MSD ANIMAL HEALTH
SCIENTIFIC BOOK
World Buiatrics Congress
July 3rd - 8th, 2016
Dublin, Ireland
© 2015 Intervet International B.V., also known as MSD Animal Health. All rights reserved.
GL / RUM / 0 516 / 0 0 0 9
INTRODUCTION
Through its commitment to the Science of Healthier Animals™, MSD Animal Health
offers ruminant veterinarians and farmers, one of the widest ranges of veterinary
pharmaceuticals, vaccines and health management solutions and services. MSD
Animal Health is dedicated to preserving and improving the health, well-being and
performance of ruminants.
To emphasize this, MSD contributes both financially to World Buiatrics Congress
as a premium partner sponsor, and scientifically through 34 posters and 11 oral
presentations. The 11 oral presentations assigned to MSD in this 2016 WBC congress,
is an outstanding performance and a company record!
To create awareness within and outside MSD for the hard work and quality of all
these studies, we have produced this book. In the book, you will not only find all
posters, but also key powerpoint slides of each oral presentation. The contributions
content list is presented as a scientific program per day of the congress. This way it
is easy to find out when and where you can visit the posters or oral presentations you
are interested in during the congress.
Leaves me to congratulate all authors with their contributions to MSD, to ruminant
science and to the WBC congress,
Jantijn Swinkels, DVM, PhD
Global Technical Director & Editor
“THE SCIENCE OF HEALTHIER ANIMALS”
MSD SCIENTIFIC PROGRAM
Day/Time/Location Title/Author(s) Page
July 4, 2016
ORAL PRESENTATIONS
3:45 PM-4:00 PM Systematic review and meta-analysis of the effectiveness of cefquinome treatment protocols of 1
Auditorium clinical mastitis in dairy cows
Lina M. González, Hernando G. Gaitán, Claudia G. Cobo, Javier Sánchez, Carlos Lüders, Alejandro Ceballos
POSTER PRESENTATIONS
8:00 AM-6:30 PM Effectiveness and safety of a novel flunixin meglumine transdermal pour-on solution in the 2
P01-001-195 treatment of bovine mastitis
3
8:00 AM-6:30 PM Julien Thiry, Gaelle Milon-Harnois, Mathieu Chiquet, Laurent Daluzeau, Monika Borchert-Stuhlträger, 4
P01-001-022 Björn Sander, Emmanuel Thomas, Albert Boeckh, Vincent de Haas, Philippe Brianceau 5
8:00 AM-6:30 PM
P01-001-194 Evolution of seroprevalence for BHV-1 gE in a French herd vaccinated with a live bovine
8:00 AM-6:30 PM herpesvirus type 1 marker vaccine
P01-001-006
René Fournier, Loic Oliviero, Siddartha Torres
Intramammary prednisolone treatment of naturally occurring clinical mastitis only increased
clinical cure in mild cases
Jantijn Swinkels, Gaelle Milon-Harnois
Digital communication and knowledge transfer for veterinary practitioners and their client animal
owners in Ireland
Bosco Cowley
July 5, 2016
ORAL PRESENTATIONS
10:30 AM-10:45 AM There is a need to raise farmers’ awareness of correct vaccine storage temperatures 6
Wicklow Hall 2 Williams, P., Paixao, G. 7
10:45 AM-11:00 AM Intranasal vaccination in the face of maternal antibody induces long-term mucosal immune 8
Wicklow Hall 2 memory
Kevin L. Hill, Philip J. Griebel
3:00 PM-3:15 PM Failure to define the aetiology of acute respiratory disease in thirteen of fifteen live dairy cows in
Liffey B Scotland
Crawshaw, W., Williams, P.
POSTER PRESENTATIONS
8:00 AM-6:30 PM Comparative time, behavior and economic differences associated with calves receiving a pour-on 9
P02-002-044 vs. an intravenous product 10
11
8:00 AM-6:30 PM David Amrine, Siddartha Torres 12
P02-002-098 13
Improving colostrum management in dairy calves by blood IgG measurement combined with
8:00 AM-6:30 PM recording and analyzing colostrum management data
P02-002-064
8:00 AM-6:30 PM Nijhoving G.H., Kuijk H.A., Simons B.A.
P02-002-179
Validation of an easy-to-use device for non esterified fatty acids assay in blood in dairy cattle
8:00 AM-6:30 PM
P02-002-135 Louise Gros, Lucile Puech, Christian Engel, Jantijn Swinkels, René Fournier, Sylvie Chastant-Maillard
Absence of association between pre and post partum energy status and genital inflammation in
dairy cows
Louise Gros, Lucile Puech, Christian Engel, Jantijn Swinkels, René Fournier, Sylvie Chastant-Maillard
Vaccination against clostridial disease in maternally derived antibody positive lambs; a
randomised, non-inferiority field trial in twin lambs
John Moffat, Hannah Bain, Stu Bruere, Jantijn Swinkels
MSD SCIENTIFIC PROGRAM
Day/Time/Location Title/Author(s) Page
8:00 AM-6:30 PM Subcutaneous ischiorectal fossa injection: serological, colostral immunity and tissue reactivity in 14
P02-002-145 rota, corona and E. coli F5 (K99) vaccinated cows 15
8:00 AM-6:30 PM 16
P02-002-146 John Moffat, Kate Golyn, Richard Smith, Jantijn Swinkels 17
8:00 AM-6:30 PM 18
P02-002-106 Does BVD vaccination of recently weaned pasture-fed beef cattle prevent weight loss when
8:00 AM-6:30 PM exposed to persistently infected cattle? 19
P02-002-034 20
8:00 AM-6:30 PM John Moffat, Stu Bruere, Jantijn Swinkels
P02-002-153 21
Evaluation of quality and management of colostrum on dairy farms in Spain
8:00 AM-6:30 PM 22
P02-002-051 Carlos Carbonell, Laura Ferreira, Laura Elvira
8:00 AM-6:30 PM
P02-002-186 Education of the dairy employee on animal care – A practical approach using Dairy CARE 365
8:00 AM-6:30 PM Michael W. Bolton, Paula Ospina, Elizabeth A. Cox, J. K. Shearer, Ben Bartlett, Fred J. Muller, Gregory J.
P02-002-182 Crosley, Scott Nordstrom
8:00 AM-6:30 PM Vaccination of dairy cows in late gestation: effect on serum and colostrum antibody titers for
P02-002-197 BRSV and Mannheimia haemolytica
Laurent Nouvellon, Vincent Tessier, Evelyne Michel, Sébastien Assie, Bertrand Ridremont, Jantijn
Swinkels
The healthy calf – A nationwide survey on the IgG-concentration in newborn calves serum
Elke Rauch, Nicola Franz, Sven Reese, Michael Erhard
Protocols combining GnRH and a prostaglandin analogue for the treatment of ovarian cysts in
dairy cows
Ana Cristina Figueiredo, Carlos Fernandes, Joao Henrique Viana, Mauricio R. Vieira, Denis A. Antonio,
Henderson Ayres, Joao Paulo Barbuio
Equine chorionic gonadotropin treatment in a fixed-time artificial insemination protocol in anestrus
beef cows
Ana Cristina Silva de Figueiredo, Carlos Antônio de Carvalho Fernandes, Henderson Ayres, João Paulo
Barbuio, Rafael Marin Chiummo, Denis Barbosa Alves Antonio
One single dose of equine chorionic gonadotropin is sufficient for superovulation of cows
Carlos Antônio de Carvalho Fernandes, Ana Cristina Silva de Figueiredo, Henderson Ayres, João Paulo
Barbuio, Rafael Marin Chiummo, Denis Barbosa Alves Antonio
July 6, 2016
POSTER PRESENTATIONS
8:00 AM-6:30 PM Field prevalence of bovine respiratory pathogens by PCR technique in France 23
P03-003-101 24
8:00 AM-6:30 PM Siddartha Torres, Vincent Tessier, Bertrand Ridremont
P03-003-213 25
Resumption of ovarian cyclicity and energy balance in Holstein cows in France based on in-line 26
8:00 AM-6:30 PM milk measurements 27
P03-003-029 28
Claire Saby-Chaban, René Fournier, Faouzi Lyazrhi, Edouard Alix, Jantijn Swinkels, Sylvie Chastant-
8:00 AM-6:30 PM Maillard
P03-003-090
8:00 AM-6:30 PM Monitoring of BVDV by testing antibodies to NS3 protein in calves vaccinated with an inactivated
P03-003-234 BVDV vaccine
8:00 AM-6:30 PM H. Kuijk, G. Nijhoving, L. van Duijn, M. Mars, W. Theuws, H. Swam, B. Makoschey
P03-003-226
Molecular epidemiology of bovine viral diarrhea virus in Spain, spatial and temporal distribution
F. Esperón, L. Elvira, A. Torre, V. Delicado, M. Álvarez
Efficacy of 86μg Gonadorelin and Cloprostenol Sodium in a dairy Fixed-Time Artificial Insemination
program
Lina S. de Montigny, Terry Katz, Paul M. Baldwin, Jantijn Swinkels
Effect of two different bovine somatotropins on milk production and reproduction of dairy cows in Brazil
Henderson Ayres, Marcilio Nichi, Sandro L. Viechnieski, Pietro Sampaio Baruselli, Rodrigo Almeida
MSD SCIENTIFIC PROGRAM
Day/Time/Location Title/Author(s) Page
July 7, 2016
ORAL PRESENTATIONS
9:15 AM-9:30 AM Comparison of Two Macrolides in the Control of BRD in Calf Ranch Cattle 29
Wicklow Hall 2 Norman Stewart 30
31
9:30 AM-9:45 AM Efficacy of flunixin meglumine pour-on administration in a tissue cage model of inflammation
Wicklow Hall 2 Olivier Roy, René Fournier, Julien Thiry, Philippe Brianceau, Siddartha Torres 32
9:45 AM-10:00 AM Early detection of bovine respiratory disease and immediate treatment with flunixin help to reduce 33
Wicklow Hall 2 the use of antibiotics in beef calves 34
35
C. C. Gelfert, C. Ströbel, C. Beisl, E. Thesing, D. Lange
11:30 AM-11:45 AM Treatment of naturally occurring bovine respiratory disease in juvenile calves with a single
Wicklow Hall 2 administration of a florfenicol plus flunixin meglumine formulation
Julien Thiry, Vincent de Haas
11:45 AM-12:00 PM Associations between lung consolidation, pneumonia treatment and live-weight in UK veal cattle
Liffey Hall 2 Williams, P., Potter, T., Cooper, R.
12:00 PM-12:15 PM On farm monitoring of the four Q’s of colostrum management is important
Liffey Hall 2 Hart, K.R., Reyher, K.K., Barrett, D.C., Williams, P.D.
12:30 PM -12:45 PM Effectiveness of tildipirosin and tulathromycin in the control of bovine respiratory disease in high
Wicklow Hall 2 risk beef heifers
Riccardo Compiani, Stefano Jottini, Siddartha Torres
POSTER PRESENTATIONS
8:00 AM-6:30 PM Flunixin meglumine transdermal pour-on solution as adjunct therapy in the treatment of bovine 36
P04-004-114 respiratory disease in calves less than 8 weeks of age 37
38
8:00 AM-6:30 PM Julien Thiry, Siddartha Torres, Vincent de Hass, Philippe Brianceau 39
P04-004-116 40
Efficacy of injectable florfenicol plus flunixin combination against an experimentally induced 41
8:00 AM-6:30 PM Mycoplasma bovis infection in calves 42
P04-004-108 43
Julien Thiry, Sophie Rubion, Cliff Ramage, David Reddick, Vincent de Haas 44
8:00 AM-6:30 PM 45
P04-004-123 Pulmonary lesions and clinical disease in Histophilus somni-challenged calves treated with, either
8:00 AM-6:30 PM tildipirosin or tulathromycin
P04-004-073
Anthony W. Confer, Jared D. Taylor, Timothy A. Snider, Mark Spire
8:00 AM-6:30 PM
P04-004-110 Incidence and antibiotic susceptibility of non-viral pathogens isolated from calves with BRD
8:00 AM-6:30 PM Nijhoving G.H., Kuijk H., Geurts C.P.H., Botvliet I.
P04-004-111
Fipronil and Fluazuron: efficacy of a novel parasiticide combination against Dermatobia hominis
8:00 AM-6:30 PM and Cochliomyia hominivorax in naturally infested cattle
P04-004-112
Lopes, W.D.Z., Chiummo, R.M., Botelho, E.M., Vettorato, L.F.
8:00 AM-6:30 PM
P04-004-113 Pharmacokinetic evaluation of a new combination of Fipronil and Fluazuron topically administered
8:00 AM-6:30 PM (pour-on) in cattle
P04-004-115
Lopes, W.D.Z., Chiummo, R. M., Botelho, E.M., Vettorato, L.F.
Therapeutic and residual efficacy of a novel Fluazuron combination against Rhipicephalus
(Boophilus) microplus ticks in naturally infested cattle
Lopes, W.D.Z., Chiummo, R. M., Botelho, E.M., Vettorato, L.F.
Comparative therapeutic and residual efficacy of a novel parasiticide combination of fipronil and
fluazuron against Rhipicephalus (Boophilus) microplus ticks in artificially infested cattle
Lopes, W.D.Z., Chiummo, R. M., Botelho, E.M., Vettorato, L.F.
Antimicrobial susceptibility of staphylococcus aureus isolated in bovine mastitis in France
Sylvain Bareille, Frédéric Leboeuf, Guillaume Lequeux, Jantijn Swinkels
Effect of two different bovine somatotropins on milk production and reproduction of dairy cows in Brazil
Henderson Ayres, Marcilio Nichi, Sandro L. Viechnieski, Pietro Sampaio Baruselli, Rodrigo Almeida
MSD ANIMAL HEALTH
Systematic review and meta-analysis of the effectiveness
of cefquinome treatment protocols of clinical mastitis in
dairy cows
Lina M. González*1, Hernando G. Gaitán1, Claudia G. Cobo2, Javier Sánchez3, Carlos Lüders4, Alejandro Ceballos2
1Universidad Nacional de Colombia, Bogotá, 2Universidad de Caldas, Manizales, Colombia, 3University of Prince Edward Island, Charlottetown, Canada, 4Universidad Católica de Temuco, Temuco, Chile
*Presenting Author
BACKGROUND
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
1 WBC 2016 (961)
MSD ANIMAL HEALTH
Effectiveness and safety of a novel flunixin meglumine transdermal
pour-on solution in the treatment of bovine mastitis
Julien Thiry1, Gaelle Milon-Harnois1, Mathieu Chiquet1, Laurent Daluzeau1, Monika Borchert-Stuhlträger2, Björn Sander2, Emmanuel Thomas2, Albert Boeckh3, Vincent de
Haas1,3, Philippe Brianceau3
1 MSD Animal Health Innovation, 49071 Beaucouzé, France, 2 MSD Animal Health Innovation, 55270 Schwabenheim, Germany, 3 MSD Animal Health, Madison, NJ USA
OBJECTIVES Rectal temperature (ºC) 41 Flunixin Pour-On
40.5 Negative Control
Flunixin has anti-inflammatory, anti-pyretic and analgesic effects. This molecule is
commonly used for the relief of pain and control of inflammation and pyrexia associated 40 FIGURE 1
with diseases of different origin and nature. A novel 50 mg/ml flunixin transdermal 39.5
formulation was developed by MSD Animal Health (Finadyne® Transdermal) and is Effect of treatment on
now the first NSAID registered to be administered as a pour-on product along the 39 mean rectal temperature
dorsal midline in cattle. The objective of the present study was to determine the field 38.5 (°C). The difference at
effectiveness of the 50 mg/ml flunixin transdermal formulation as adjunct therapy in the 6 hours is statistically
treatment of bovine mastitis. 38 24 48 72 96 120 144 significant (p<0.0001).
0 Time post-treatment (hours)
MATERIALS AND METHODS
A B
A total of 133 animals located in France, Germany and Spain, aged from 2 to 12 years,
and showing acute signs of mastitis as evidenced by at least two of the clinical signs 80% 80%
of udder inflammation (swelling, pain, firmness) scored as moderate or severe; milk
modified or highly modified; and rectal temperature ≥ 40.0°C, were enrolled by 13 70% 70%
Investigators at 48 dairy farms and and randomly assigned to treatment. 60% 60%
Each animal was treated either with the test product, 50 mg/ml flunixin (Finadyne®
Transdermal; 3.3 mg/kg flunixin; 1ml/15kg; MSD Animal Health) administered topically Percentage of Animals 50% 50% Percentage of Animals
once along the dorsal midline, or with a negative control product, both administered
once topically along the dorsal midline. At 6 hours post-treatment administration and on 40% 40%
day 1, animals received a systemic antibiotic (Cobactan® 2.5%; 1 mg/kg cefquinome;
2ml/50kg; MSD Animal Health) administered by intramuscular injection. At 6 hours post- 30% 30%
treatment, following the final efficacy evaluation, an intrammary antibiotic (Cobactan®
LC, 75 mg cefquinome per syringe; MSD Animal Health) was also administered by 20% 20%
infusion into the teat of each affected quarter and every 12 hours after each of 3
successive milkings. To avoid observation bias and to preserve masking, treatments 10% 10%
were dispensed and administered by individuals not involved in the clinical assessment.
All animals were observed for clinical signs of disease for 6 hours following treatment 0% -2 -1 0 12 3 PImaipnroSvceomreent 0% -1 0 123 IPmaipnroSvceomreent
initiation and daily thereafter for 6 consecutive days. The dosing sites was evaluated and
any adverse effects were recorded. Treatment Finadyne Transdermal Negative Control Treatment Finadyne Transdermal Negative Control
All animals had a milk sample collected for bacterial culture, identification and
determination of cefquinome sensitivity. FIGURE 2
RESULTS Percentage of individual pain score improvement between day 0 enrollment and day 0 at 6 hours (A), and between
day 0 enrollment and day 1 (B). 31 animals in the flunixin group (31/64 = 48.44%) had at least a 1 point decrease in
A total of 129 pathogens were isolated on day 0 prior to treatment and 14 pathogens pain score between enrollment and 6 hours post-treatment (A). 43 animals in the flunixin group (43/55 = 78.18%) had
were isolated post-treatment from animals classified as treatment failure. Escherichia at least a 1 point decrease in pain score between enrollment and 24 hours post-treatment (day 1) (B).
coli (n=49 or 38.0%) and Streptococcus uberis (n=43 or 33.3%) were the most
prevalent pathogens at enrollment and were isolated at almost all sites. A very high 100% 21.21% Normal Mild Moderate Severe 3.85%
number of isolates (95.7%) were found to be susceptibe to cefquinome. 90% 10.58% 9.38%
Both treatments were effective to reduce pyrexia; however, the decrease in temperature
was significantly greater with flunixin transdermal (-1.8°C) compared to the negative 80% 15.63%
control group (-1°C) (Fig. 1). This difference was statistically significant (p<0.0001).
Flunixin transdermal acted quickly and was efficacious to prevent the deleterious effects 70% FIGURE 3
of inflammation as it was shown by the udder pain, firmness and swelling improvement
at 6 hours post-treatment and on day 1 (Fig. 2, 3, 4, 5 and 6). 60% 39.39% 89.42% 51.56% 46.97% Udder pain score (%) at enrollment and at
No treated animal presented dosing site reactions throughout the observation period. 50% 6 h post-treatment.
Neither flunixin nor control had a negative influence on the health status including
appetite and faecal consistency. 40%
30%
20% 30.30% 31.34% 23.44% 33.33%
9.09%
10%
0% 9.09% 4.48%
Flunixin Transdermal Negative Control Flunixin Transdermal Negative Control
Enrollment (Time 0h) 6 hours post-treatment (Time 6h)
A B
90% 90%
80% 80%
70% 70%
Percentage of Animals 60% 60%
Percentage of Animals
50% 50%
Percentage of Animals
Percentage of Animals40% 40%
30% 30%
20% 20%
10% 10%
0% -1 0 1 2 FImirpmrnoevessmSecnotre 0% -1 0 1 2 FImirpmrnoevsesmSecnotre
A B Treatment Finadyne Transdermal Negative Control Treatment Finadyne Transdermal Negative Control
90% 90% FIGURE 4
80% 80% Percentage of individual firmness score improvement between day 0 enrollment and day 0 at 6 hours (A), and
between day 0 enrollment and day 1 (B). 23 animals in the flunixin group (23/64 = 35.94%) had at least a 1 point
70% 70% decrease in pain score between enrollment and 6 hours post-treatment (A). 35 animals in the flunixin group (35/55 =
63.64%) had at least a 1 point decrease in firmness score between enrollment and 24 hours post-treatment (B).
60% 60%
50% 50% 100% Normal Mild Moderate Severe
90% 20.90% 17.19%
40% 40% 33.44% 27.27%
30% 30% 80%
20% 20% 70% FIGURE 5
60% 57.81% 66.67% Udder firmness score (%) at enrollment
and at 6 h post-treatment.
10% 10% 50%
0% -2 -1 0 1 2 ISmwperollvinegmSecnotre 0% -1 0 1 2 3 SImwperollvinegmSecnotre 40% 63.64% 73.13%
30%
Treatment Finadyne Transdermal Negative Control Treatment Finadyne Transdermal Negative Control 20%
FIGURE 6 10% 25.00% 6.06%
0% 3.03% 5.97%
Percentage of individual swelling score improvement between day 0 enrollment and day 0 at 6 hours (A), and between Flunixin Transdermal Negative Control Flunixin Transdermal Negative Control
day 0 enrollment and day 1 (B). 25 animals in the flunixin group (25/64 = 39.06%) had at least a 1 point decrease in 6 hours post-treatment (Time 6h)
swelling score between enrollment and 6 hours post-treatment (A). 39 animals in the flunixin group (39/55 = 70.91%) Enrollment (Time 0h)
had at least a 1 point decrease in swelling score between enrollment and 24 hours post-treatment (B).
Summary of efficacy results
100% Normal Mild Moderate Severe Variable Flunixin Control
90% 22.39% 10.94% N enrolled animals 66 67
80% Age (days) at enrollment
31.82% 27.27% Body weight (kg) at enrolment 4.6 ± 1.9 4.8 ± 2.2
Rectal temperature (°C) at enrollment 668.2 ± 67.4 672.9 ± 66.0
70% FIGURE 7 Rectal temperature (°C) at 6 h after treatment 40.4 ± 0.4
Drop in temperature 6 h after treatment * 38.6 ± 0.4 40.6 ± 0.6
60% Udder swelling score (%) at enrollment and Udder inflammationa at enrollment 39.6 ± 0.7
at 6 h post-treatment Udder inflammationa at 6 h after treatment -1.8 °C
50% 62.12% 70.15% 62.50% 62.12% 6.3 ± 1.3 -1 °C
40% TABLE 1 4.9 ± 1.5 6.0 ± 1.1
6.0 ± 1.4
30% Summary of animal demographics and efficacy results
20%
10% 4.55% 4.48% 25.00% 10.61%
0% 1.52% 0.00% 1.56% 0.00%
Flunixin Transdermal Negative Control Flunixin Transdermal Negative Control
Enrollment (Time 0h) 6 hours post-treatment (Time 6h)
CONCLUSIONS
Based on the study results, it can be concluded that the new 50 mg/ml flunixin transdermal solution had a significant positive effect on
clinical signs that are very relevant to the assessment and monitoring of animals affected by mastitis and how they overall respond to
treatment. Finadyne® Transdermal 50 mg/ml pour-on solution for cattle has strong anti-pyretic effect and anti-inflammatory properties,
and make it a very convenient and suitable adjunct therapy to anti-infective therapy used in cases of mastitis in cattle.
MSD Animal Health 2
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P01-001-195)
MSD ANIMAL HEALTH
Evolution of seroprevalence for BHV-1 gE in a French
herd vaccinated with a live bovine herpesvirus type 1
marker vaccine
René Fournier1, Loic Oliviero1, Siddartha Torres2
1MSD Santé Animale, Beaucouzé, France, 2Merck Animal Health, Madison, United States
INTRODUCTION Seroincidence 7% FIGURE 1
5%
A herd vaccination programme was 4% IBR gE Seroincidence
started in November 2008 in a large 2%
beef herd in Normandy with a high Animals with
seroprevalence of IBR (Infectious Seroconversion /
Bovine Rhinotracheitis), due to Bovine
Herpesvirus type 1 (BHV-1) antibodies. tested animals
Seventy-four % of heifers (88/119),
at 14 to 30 months of age, that were
vaccinated with a live marker vaccine
(Bovilis® IBR Marker Live, MSD Santé
Animale) had BHV-1 gE antibodies before
beginning the herd protocol.
MATERIALS AND METHODS 7% 2009 Nov. 2010 2011 Nov. 2012
2010 May 2011 2012 May 2013
All cattle within this herd, n=400, 50% Nov. 2013
including calves, were vaccinated with 38% Nov. Nov. May 2014
the live marker vaccine as mentioned 25% May May Nov. 2014
above. Calves were vaccinated intra- 13%
nasally at two weeks of age, revaccinated Seroprevalence ELISA gE Serological testing dates
at three months of age and thereafter 0%
boosted every six months. Cattle aged FIGURE 2
3 months or more were vaccinated May 2009 NMoav.y 22000109 2010 2011 2012 2013 Nov. 2014
once intramuscularly and revaccinated 2011IBR gE Seroprevalence2012 2013 2014
every six months. All the animals aged
between 12 and 48 months were Nov. Seropositive
blood-sampled every six months to May animals /
evaluate their BHV-1 gE antibody status.
Biosecurity measures were also applied tested animals
and every gE antibody positive animal
was removed from the herd. ELISA gE Serological testing dates
Nov. Nov. Nov.
RESULTS May May May
The incidence of new seroconversion
(seroincidence) decreased from 5 % in
November 2009 to 0 % in November
2012 (Fig. 1). As a result, the herd
seroprevalence decreased from 50 % in
May 2009 to 0 % in May 2014 (Fig 2.).
CONCLUSION
It can be concluded that, under field
conditions, vaccination with a live IBR
marker vaccine could significantly
contribute, if combined with strict
biosecurity measures, to the eradication
of BHV-1 virus from a large herd with a
high seroprevalence.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
3 WBC 2016 (P01-001-022)
MSD ANIMAL HEALTH
Intramammary prednisolone treatment of naturally occurring
clinical mastitis only increased clinical cure in mild cases
Jantijn Swinkels1, Gaelle Milon-Harnois2
1MSD Animal Health, Global Ruminants, Boxmeer, The Netherlands, 2MSD Animal Health, Global Statistics, Beaucouzé, France
INTRODUCTION color, and dry cow treatment. A P>0.10 was used to exclude variables. The
interaction of these selected potential confounding variables with treatment
In the EU, the majority of dairy cows with clinical mastitis are intramammary treated was then assessed. Parameters were considered significant using P<O.05.
with prednisolone assuming alleviation of clinical symptoms. Experimental studies
show a reduction of clinical signs by preventing the overshooting of the immune- RESULTS
system, when treatment is a few hours after challenge of LPS or E. coli. The question
is whether these effects can be translated to the field. 113 cases were included, 55 in Group 1 and 58 in Group 2. Most post-
admission withdrawals were due to protocol deviations, i.e. mastitis in a
OBJECTIVE second quarter (n = 23), off protocol treatment during the trial period (n=1).
To determine the clinical efficacy of prednisolone in naturally occurring clinical mastitis Being a Holstein Friesian, pre-dipping teats and having mild or moderate vs,
cases under field conditions. severe initial udder swelling, were significantly associated with increased
odds of a clinical cure (Table 2). In the final model, only the interaction
MATERIAL AND METHODS between treatment and initial udder swelling was significant, the addition of
prednisolone to cefapirin significantly increased the odds of a clinical cure in
The multi-centered, double-blinded, randomized, and controlled trial was performed cases of mild mastitis (Table 3).
in France according to GCP guidelines. 120 cows with naturally occurring mastitis in a
single quarter were randomly (SAS® software generated list) allocated to one of two
treatment groups.
TREATMENT TABLE 2.
Group 1: 300 mg of cefapirin intramammary, 2 x per day for 2 consecutive days. Significant (P<0.05) potential confounding variables associated with clinical
cure of clinical mastitis modelled using logistic regression.
Group 2: 300 mg of cefapirin plus 20 mg prednisolone, 2 x per day for 2 consecutive
days. Effect Estimate SE t-value P-value OR 95% CI
Bacteriological analysis was performed in a centralized lab (IDEXX) only on pre- Intercept -1.545 0.71 4.73 0.03
treatment milk samples of the affected quarter (not reported here). Clinical
examinations were performed as shown in Table 1. Breed
Holstein Fresian vs
other 0.822 0.34 5.84 0.02 5.17 1.36 – 19.6
Pre-dipping
TABLE 1.
Schedule of the trial and the clinical evaluations.
Day Day 0 Day 1 Day 2 Day 3 Day 4 Day 5 Yes vs no 0.868 0.32 7.55 0.006 5.68 1.65 – 19.6
Time of AM PM AM PM AM PM AM PM AM PM AM PM Rectal temp D0
examination (milking)
Clinical evaluation XXXX X X ≤39.5 vs > 39.5 1.216 0.63 3.69 0.055 11.38 0.95 – 136.1
by Investigator
Clinical evaluation X X X X X X Severity D0
by Breeder
Mild vs severe 1.085 0.45 5.92 0.01 14.20 1.84 – 109.9
Inclusion X
Allocation to X Moderate vs severe 0.484 0.40 1.47 0.22 7.79 1.13 – 53.64
treatment group
Milk sampling for X TABLE 3.
bacteriology
Final logistic regression model of clinical cure of clinical mastitis comparing
Treatment T1 T2 T3 T4 treatment with 300 mg cefapirin vs 300 mg cefapirin with 20 mg of
prednisolone. Prednisolone increased the odds of cure in mild mastitis, yet
CLINICAL PARAMETERS decreased the odds in moderate mastitis.
Each of the following parameters was clinically scored as 0= normal, 1= mild, Effect Odds ratio 95% CI
2= moderate, 3= severe: general condition (appetite, apathy, lying down), udder
firmness, udder induration and udder pain (all by palpation), and milk characteristics Mild mastitis
(colour, flocks, flakes).
Prednisolone yes vs no 3.69 1.05 -12.90
A cow was normal if clinical signs of mastitis were absent, all parameters above
scored 0, and the rectal temperature was <39.5°C. Moderate mastitis
DEFINITION OF CURE Prednisolone yes vs no 0.23 0.07 - 0.78
Because farmers usually judge efficacy at the end of label treatment, clinical Severe mastitis
efficacy was judged 1 day after the last treatment, on day 3. A case was
considered cured at this time, if based on the clinical parameters above, she was Prednisolone yes vs no 1.17 0.25 - 5.59
deemed normal. Severe cases were defined as a sum of scores ≥ 5 and at least
2 scores ≥ 2, OR sum of scores ≥ 5 and temperature ≥ 39.5°C. Moderate cases DISCUSSION AND CONCLUSION
were defined as sum scores ≤ 5 and no more than one score ≥ 2. A case with a
normal general condition, a sum score ≤ 4 and no scores > 1, was considered a Evidence based medicine is a pre-requisite in modern agriculture. To the
mild case. authors’ knowledge, this is the first study looking at the clinical effects
of intrammammary prednisolone treatment of clinical mastitis in the
BLINDING field. Although many veterinarians perceive anti-inflammatory only to be
necessary in moderate or severe clinical mastitis cases, this study shows
Treatment was performed by the investigator. Both treatments were labelled prednisolone treatment only increased clinical cure when symptoms were
similarly and could not be distinguished from each other, so neither the investigator mild.
nor the farmer were aware of the treatment.
Mild symptoms may reflect a well-functioning immune system or the early
STATISTICAL ANALYSIS stages of infection where prednisolone has the opportunity to prevent
immune system ‘overshooting’ and be more effective in preventing tissue
Logistic regression was used to model the association between clinical cure and damage. If this is true, early treatment with prednisolone is warranted to
treatment. A stepwise-backwards selection method, was performed to assess ensure clinical efficacy.
the association of the following potential confounding variables: farm-level:-farm
size, housing, pre- and post-milking teat disinfection, and BMSCC: cow-level:-
breed, days in milk, milk yield, individual (log) SCC, mastitis in current and previous
lactation, initial clinical score, severity, rectal temperature, pathogen, Gram
MSD Animal Health 4
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P01-001-194)
MSD ANIMAL HEALTH
Digital communication and knowledge transfer for veterinary
practitioners and their client animal owners in Ireland
DJ Bosco Cowley1
1MSD Animal Health, Red Oak North, South County Business Park, Leopardstown, Dublin 18, Ireland
INTRODUCTION FIGURE 1
Digital devices and channels are becoming more critical to effective knowledge transfer, Percentage of ownership of electronic devices by vets (n=350) and cattle farmers (n=385) in Ireland
principally due to perceived improved cost-effectiveness, efficiency and “always-on”
availability. To address a dearth of publications on the evolution and uptake of digital FIGURE 2
technologies, regarding Registered Veterinary Practitioners (RVPs) in Ireland, and on their
bovine client animal owners (CAOs), the aims of this research are to: Ranking of importance of information sources for vets (n=350) in Ireland
• Compare adoption of digital technology devices among RVPs and cattle owners to FIGURE 3
general population survey data
Ranking of importance of information sources for vets (n=350) in Ireland
• Evaluate digital communication and education by the RVPs and their CAOs
FIGURE 4A FIGURE 4B
• Determine the relative importance of information sources for these segments
% of veterinary practices with websites by county % of veterinary practices with a Facebook page by
• Evaluate patterns of their online behaviour regarding animal healthcare county
MATERIAL AND METHODS
A paper-based survey on uptake of digital devices, communication and online behaviour
concerning sources of information and education was conducted by MSD Animal Health
(MSD AH) Ireland, of RVPs, by convenience sampling at ten veterinary meetings in Ireland
between November 2014 and March 2016. Cattle farmers (CAOs) were also targeted with
a paper-based survey, comprising the same topics relevant to farmers, at eleven meetings
conducted by MSD AH and other organisations (Teagasc), between July 2015 and March
2016. RVP uptake of websites and social media was evaluated through an extensive
search engine trawl of all veterinary websites in Ireland. Digital adoption, communication
and online behaviour data were gathered from the surveys of the target personas and
the general population for comparison. Survey results were statistically analysed using a
statistical software package (STATA®) and tabulated.
RESULTS
Three hundred and fifty RVP and three hundred and eighty five CAO surveys were
collected. 78% of RVPs own a laptop, with 79% owning a smartphone and only 45%
owning a tablet device (Fig. 1). Whilst almost 50% of vets use search engines weekly,
colleagues, journals and conferences ranked in descending order as information sources
(Fig. 2). 82% of cattle farmers owned a smartphone, 61% a laptop and only 31% a tablet
device (Fig. 1). Smartphones were more or less equally used for email, telephony, text and
internet surfing but less for social media (Fig. 3). 52% of cattle farmers read their emails
every day and an additional 24% a few times per week. Daily social media usage by cattle
farmers was significantly lower than the general population (35% for farmers compared to
67% in the Irish population) (IPSOS-MRBI, 2015).
Website ownership or social media activity by RVPs ranged between 50-80% varying by
county (Figs. 4 a & b), significantly associated with practice size (p<0.02). Facebook usage
was correlated to presence of a website. Overall, 37% of veterinary practices had a website
while 48% had a Facebook page. These align well with Irish data for website presence in
small and medium enterprises (Kennedy, 2013) though uptake is lower than across all Irish
enterprises for both websites and social media (Eurostat, 2015). Websites were used to
market the practice through organic search, but also to educate their owners on animal care,
more commonly for companion animals than ruminants (25% compared to 17%). RVPs
most commonly communicated information to clients via SMS text followed by hard copy
and email (28% compared to 18% and 15% respectively). Only one third of cattle RVPs
communicated with their clients via social media compared to 48% for all Irish RVPs.
88% of cattle owners viewed the RVP as the most importance source of animal health
(AH) information with the Irish Farmers Journal (IFJ) considered the next most important.
The IFJ was the most frequently used website for seeking AH information after Google
(30% and 57% respectively). Farmers significantly preferred email over text, phone or mail
as the communication method from their vet for herd health information, though few read
e-newsletters.
CONCLUSIONS
The RVP has a significant role to play as a trusted educator on herd health. To capitalise on
this position, they must consider the most appropriate means of dissemination of credible
material across the preferred device and channel. Email may replace text as the preferred
communication tool for farmers as the smartphone dominates. For successful information
transfer, media must encourage a higher engagement rate with content than currently
exists with e-newsletters.
ACKNOWLEDGEMENTS
A sincerest thank you to Mr. Seamus Maguire, General Manager, MSD Animal Health
Ireland for sponsoring the conduct of this study and to those who assisted in collecting
surveys including members of the veterinary technical team of MSD Animal Health (Ms.
Joanne Cregg, Ms. Sharon Magnier, Ms. Mairead O’Grady, Ms. Catherine O’Leary) and Dr.
Tom O’Dwyer, Teagasc.
REFERENCES
EUROSTAT. 2015. Enterprises’ presence on the internet [Online]. Eurostat. Available at:
http://ec.europa.eu/eurostat/statistics-explained/index.php/File:Enterprises%27_presence_
on_the_internet,_2015,_(%25_of_enterprises).png.
IPSOS-MRBI. 2015. Social networking quarterly survey - August 2015. Available at: http://
ipsosmrbi.com/wp-content/uploads/2015/11/SN_Aug15.gif.
KENNEDY, S. 2013. Survey of SME Tax Payers 2013 [Online]. Available at: www.revenue.ie/
en/about/publications/business-survey-2013.pdf.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
5 WBC 2016 (P01-001-006)
MSD ANIMAL HEALTH
There is a need to raise farmers’ awareness of correct vaccine
storage temperatures
Williams, P.*1, Paixao, G.2
1MSD Animal Health, Walton Manor, Walton, Milton Keynes, MK7 7AJ UK, 2University of Bristol, Langford House, Langford, Bristol, BS40 5DU UK
*Presenting Author
MSD Animal Health 6
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (IV-027-006)
MSD ANIMAL HEALTH
Intranasal Vaccination in the Face of Maternal Antibody
Induces Long-term Mucosal Immune Memory
Kevin L. Hill*1, Philip J. Griebel2
1Merck Animal Health, Desoto, United States, 2VIDO/Intervac, Saskatoon, Canada
*Presenting Author
MSD Animal Health
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7 WBC 2016 (IV-027-007)
MSD ANIMAL HEALTH
Failure to define the aetiology of acute respiratory disease in
thirteen of fifteen live dairy cows in Scotland
Crawshaw, W.*1, Williams, P.2
1MBM Veterinary Group, Kilmarnock, KA3 1HA, UK, 2MSD Animal Health, Walton Manor, Walton, Milton Keynes, MK7 7AJ, UK
*Presenting Author
MSD Animal Health 8
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (ED-026-002)
MSD ANIMAL HEALTH
Comparative time, behavior and economic differences associated
with calves receiving a pour-on vs. an intravenous product
David Amrine1, Siddartha Torres2
1Professional Beef Services, Manhattan, Unites States, 2Merck Animal Health, Madison, United States
INTRODUCTION RESULTS AND DISCUSSION
Intravenous administration requires proper animal restrain, familiarity with bovine Average body weight (weight, ± standard deviation) was 268 ± 25.2kg for IV treated
anatomy, experience, and it potentially increases time and costs compared to pour- calves and 259 ± 30.0 kg for POUR treated calves.
on administration. Also, it requires appropriate fixation of the animal’s neck and
head, and it can be associated with increased handling stress of the animal. Time
Least squares mean total time in the head gate was lower (P = 0.02) for POUR
OBJECTIVE calves (seconds, ± standard error, 17.2 ±1.25) compared to IV (53.6 ±1.24) treated
calves (Figure 1, Table 3).
The goal of this study was to evaluate the differences in time, behavior, and costs Behavior
associated with pour-on or injectable administration of saline to cattle. The probability of receiving a normal vocalization score was higher (P < 0.05) in
animals receiving POUR treatment (98%) compared with those receiving IV (86%).
MATERIALS AND METHODS In addition, the probability of receiving normal chute score was higher (P < 0.05) for
calves receiving POUR treatment (34%) compared with those receiving IV (14%). The
One hundred beef crossbred calves were selected for the study. Calves were probability of having a normal exit score was numerically higher in calves receiving
randomized to treatment based on chute order. Each calf was moved into a POUR (44%) compared with those receiving IV (29%); however, this difference was
processing chute and restrained using a head gate. Calves were then allocated not significant (P = 0.11) ,Table 3 and Figure 2.
to receive either: 1) Pour-on (POUR), calves received 18 ml of sterile saline on Economic analysis
their back; or 2) Intravenous injection (IV), cattle to receive ten mL of sterile saline Least squares mean total cost of treatment administration was lower (P = 0.01)
intravenously injected by trained animal care personnel; this IV treatment required for calves receiving POUR treatment (US Dollars ± standard error, $0.72 ± $0.02)
additional restrain using a halter to fix animal head to the chute. compared to those receiving IV administration ($1.98 ± $ 0.02), Table 4.
TIME TO COMPLETE TREATMENTS AND CONCLUSIONS
MEASUREMENT OF BEHAVIORS
The pour-on administration provided a total cost savings of $1.24 ($0.54 + $0.70)
The total time was recorded using a stopwatch. Total time in the head gate for per animal compared to an intravenous injection. Animal vocalization and excessive
calves in the IV group included the time required to place a halter, restrain the movement during processing at the chute has been associated with calves
animal’s head, and properly position the animal’s neck prior to intravenous saline experiencing stressful conditions 1, 2, 3. Data from the present study indicate that
administration. Three scoring systems (Table 1) were used to capture potential calves receiving a pour-on administration were less stressed while in the chute
changes in behavior associated with treatment administration. compared to those receiving intravenously administered saline.
ECONOMIC DIFFERENCES BETWEEN TREATMENTS REFERENCES
Costs associated with labor and supplies necessary for intravenous administration 1. Vetters, M. et. al. 2012. J. Anim. Sci. 91:374381.
at a farm are displayed in table 2. The cost of a needle and syringe was estimated
for each IV administration (Table 2). 2. Curley, K. et. al., 2006, J. Anim. Sci. 84:31003103.
STATISTICAL ANALYSIS 3. Bristow, D. et. al., 2007, Physiology & Behavior 90, 626628.
Generalized linear models were used to evaluate potential differences between
treatment methods (POUR & IV) and the variables of interest.
TABLE 3. Statistical analysis of time in the head gate and behavior
scores between beef calves administered a pour-on (POUR) or intravenous (IV)
saline in NE Missouri.
TABLE 1. Behavior scoring systems
TABLE 2. Estimated costs of labor and supplies associated with TABLE 4. Estimated cost per animal (time, labor, and supplies)
administration of pour-on and IV saline to 100 beef calves in NE Missouri associated with administration of pour-on (POUR) or intravenous (IV) saline to
beef calves in NE Missouri.
(A) (B) (C)
1 0.6 0.7
0.5 0.6
0.9 0.4 0.5
0.3 0.4
0.8 0.2 0.3
Probability of no vocalization 0.1 0.2
(%) 0.1
Probability of normal chute score 0
(%)
Probability of normal exit score
(%)
0.7 0
Treatments with unlike subscripts are different (P < 0.05) POUR IV POUR IV POUR IV
Error bars represent standard error of the mean
FIGURE 2. Probability of (A) No vocalization by treatment group, (B) Normal chute score by treatment group, (C) Normal exit score by
FIGURE 1. Least squares mean time in the head gate
treatment group. Error bars represent 95% CI of the probability.
between calves receiving a pour-on (POUR) vs. intravenous (IV)
administration of saline.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
9 WBC 2016 (P02-002-044)
MSD ANIMAL HEALTH
Improving colostrum management in dairy calves by blood IgG
measurement combined with recording and analyzing colostrum
management data
Nijhoving G.H.*1, Kuijk H.A.1, Simons B.A.2
1MSD Animal Health, Boxmeer, the Netherlands, 2 Veterinary Clinic Wolvega, Wolvega, the Netherlands
* Corresponding author: [email protected]
OBJECTIVES In the low IgG blood level group of calves, a trend was seen that the amount
of colostrum in the first feeding (Figure 3) was lower than the high IgG blood
Neonatal Calf Diarrhea (NCD) is a common disease affecting newborn level group (OR=2.8, p=0.07, 95% ci 0.9-8.8) and the interval between first-
calves. To reduce the incidence of NCD it is important to use a preventive second feeding was longer than the high level IgG group (OR=2.6, p=0.09,
approach. Vaccinating the dam is a proven preventive method to increase 95% ci 0.9 -7.9).
specific ’protection’ and good colostrum management is a crucial element
for a newborn calf to get a good start. The aim of the study is to gain more There were no differences between calves with high and low IgG blood
insight in the correlation between colostrum management and blood IgG level in the interval birth-first feeding (OR=1.2, p=0.78, 95% ci 0.3-4.9),
levels of newborn dairy calves. the interval second-third feeding (OR=1.3, p=0.7, 95% ci 0.4-4.6) and the
amount of colostrum fed in the second and third feeding (OR=1.1, p= 0.85,
MATERIALS AND METHODS 95% ci 0.4-3.4), (OR=1.2, p=0.75, 95% ci 0.3-4.5).
On 13 Dutch dairy farms the colostrum management of 58 calves was CONCLUSIONS
recorded on a form during the first 24 hours (Figure 1). These calves were
blood sampled between the 2nd and the 5th day after birth to determine Conclusion of this study is that most calves still don’t receive enough
blood IgG level. colostrum. Most calves with a low blood IgG level received colostrum
directly after birth, but a strong trend was seen that the amount of
RESULTS colostrum in the first feeding was not enough and the second feeding was
offered too late. Not all farmers milked out the dam completely directly after
Sixty-two percent of the examined calves had a low blood IgG level (IgG < birth, which raises concerns.
15 g/L), the remaining 38% had a high blood IgG level (>15 g/L, Figure 2).
This study emphasized that by recording and analyzing colostrum
Most calves with a low blood IgG level received 5 liters colostrum or less management data in combination with determining the blood IgG level of
in the first 24 hours, which was significantly lower compared to the calves dairy calves, a customized advice can be given.
with a high blood IgG level (>15 g/L)(odds ratio (OR) = 5.2, p= 0.0175, 95%
confidential interval (ci) 1.3-21.5) (Figure 4).
% of calves 40%
35%
30% 5 till 9 10 till 14 15 till 19 20 till 24 >25
25%
20%
15%
10%
5%
0%
<4
Blood IgG level category (g/L)
FIGURE 1. FIGURE 2.
The colostrum management form used in this study to record data. Percentage of calves (n=58) and their blood IgG level category (g/L) between the 2nd and the 5th day after birth.
40%% of Calves % of Calves40%
35% 35%
30% 30%
25% 25%
20% 0-15 lgG g/l 20% 0-15 lgG g/l
15% 16-30 lgG gr/l 15% 16-30 lgG gr/l
10% 10%
5% 5%
0% 0%
0-0,5 0,5-1 1-1,5 1,5-2 2-2,5 2,5-3 3-3,5 3,5-4 4-4,5 4,5-5 0-1 1-2 2-3 3-4 4-5 5-6 6-7 7-8 8-9 9-10
liters liters
FIGURE 3. FIGURE 4.
Percentage of calves (total n=58) with low (green bars) and high (gray bars) IgG bood levels and their Percentage of calves (total n=58) with low (green bars) and high (gray bars) IgG blood levels and their
associated amount of colostrum (liters) category provided at the first administration after birth. associated amount of colostrum (liters) category provided during the first 24 hours after birth.
MSD Animal Health 10
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P02-002-098)
MSD ANIMAL HEALTH
Validation of an easy-to-use device for non esterified
fatty acids assay in blood in dairy cattle
Louise GROS1, Lucile PUECH1, Christian ENGEL2, Jantijn SWINKELS3, René FOURNIER4, Sylvie CHASTANT-MAILLARD1
1UMR INRA/ENVT 1225 Host-pathogen interactions Reproduction, Ecole Nationale Vétérinaire de Toulouse, France, 2Clinique vétérinaire de la Rance, Broons, France,
3MSD Animal Health, Boxmeer, The Netherlands, 4MSD Animal Health, Angers, France
CONTEXT OBJECTIVE MATERIALS AND METHODS
The transition period is key for optimal Validate a relatively cheap and easy to use 6 herds in Brittany (France)
production and health in dairy cows. device, allowing NEFA measurement by vet
100 Holstein cows
Blood concentration of non esterified practitioners
fatty acids (NEFA) is a valuable parameter between 14 days before calving to
(instead of sending samples to a lab) 14 days after calving
to evaluate its quality.
100 serum samples
RESULTS MEASUREMENT RANGE REFERENCE METHOD TESTED DEVICE
0.04 TO 2 MMOL/L
INTRA ASSAY COEFFICIENT Automated colorimetry Spectrophotometry
4.35%
RANDOX kit (London, UK) (Vetphotometer, Diaglobal
GmbH, Berlin, Germany)
COST PER ASSAY CORRELATION (SPEARMAN)
3.6 € R2=0.90; P<0.001
INTER ASSAY COEFFICIENT Reference method
17.8% LVD 14
In 4 out of 5 replicates, VetPhotometer
Assay CV%
1 13.61 [NEFA]REF = 1.7209 X [NEFA]VETPHOTOMETER + 0.0018
2 8.97
3 9.82
4 29.54
5 6.08
Mean CV 17.81%
TEST DURATION COST for the
20 minutes spectrophotometer
620€
Threshold above Time for assay Threshold Threshold
which is observed Reference Reference method VetPhotometer
Deterioration of Before calving 0.3 mmol/L 0.17 mmol/L
genital health (3-14 days before)
Chapinal et al. 2011; Ospina et al. 2013 0.4 mmol/L 0.23 mmol/L
Negative energy
balance Kaufmann et al. 2010 0.6 mmol/L 0.34 mmol/L
Deterioration of Before calving 0.7 mmol/L 0.41 mmol/L
genital health (3-14 days before)
Oetzel, 1994
Negative energy After calving
balance (3-14 days after)
Ospina et al. (2013)
After calving
(3-14 days after)
Ospina et al. (2013)
CONCLUSION
The tested Vet-Photometer device proved to be a reliable, simple, quick and low cost method with limited specific
equipment requirements for NEFA essay in the field by vet practitioners. The same device also allows Ca, Mg and
lactate blood assay.
MSD Animal Health
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11 WBC 2016 (P02-002-179)
MSD ANIMAL HEALTH
Absence of association between pre and post partum energy
status and genital tract inflammation in dairy cows
Louise GROS1, Lucile PUECH1, Christian ENGEL2, Jantijn SWINKELS3, René FOURNIER4, Sylvie CHASTANT-MAILLARD1
1UMR INRA/ENVT 1225 Host-pathogen interactions Reproduction, Ecole Nationale Vétérinaire de Toulouse, France, 2Clinique vétérinaire de la Rance, Broons, France,
3MSD Animal Health, Boxmeer, The Netherlands, 4MSD Animal Health, Angers, France
CONTEXT OBJECTIVE MATERIALS AND METHODS
Negative energy balance, both Evaluate the value of several markers 10 herds in Brittany (France)
pre- and post-partum, is one of the main of negative energy balance, of easy use
in practice, for the prediction of genital 133 Holstein cows
risk factors for genital inflammation. between 14 days before calving
Early detection of cows or herds at risk inflammation
to 14 days after calving
for negative energy balance can be
used for monitoring the quality 3-14 days before calving C 3-14 days after calving 21-35 days after calving
of the transition period A
BCS L BCS CYTOLOGY
BCS: Body Condition Score [BHB] V [BHB]
on a 0-5 scale [NEFA] I [NEFA] 1. Cervix
N 2. Uterus
G
[BHB] [NEFA] Genital cytology
β hydroxybutyrate Non Esterified Cytobrush sampling
May Grünwald Giemsa
blood Fatty Acids
concentration blood (555, RAL)
200 cells counted
Optium Xceed, concentration
Abbott Endometritis: > 5% neutrophils
VetPhotometer, Cervicitis: >6% neutrophils
Diaglobal
endomGeentriittaisl inAflNaDm/mORaticoenr:vicitis
RESULTS Energy balance Before After Difference
calving calving After - Before
Genital tract
inflammatory status D21-35 BCS BCS BCS
[NEFA] [NEFA] [NEFA]
[BHB]
ENERGY BALANCE
NO ASSOCIATION
GENITAL INFLAMMATION
Severity Presence/absence
% Endometritis
neutrophils Cervicitis
on smear
Genital inflammation
uterus
cervix
CONCLUSION
The energy balance monitoring parameters used in this study (BCS, BHB, NEFA) were not associated with the
prevalence nor by the severity of genital inflammation. Relationship between the various parameters evaluating
negative energy balance and further fertility is under investigation.
MSD Animal Health 12
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WBC 2016 (P02-002-064)
MSD ANIMAL HEALTH
Vaccination against clostridial disease in maternally derived antibody
positive lambs; a randomised, non-inferiority field trial in twin lambs
John Moffat*1, Hannah Bain1; Stu Bruere2, Jantijn Swinkels3
1MSD Animal Health, New Zealand; 2Vet Service Hawkes Bay, Masterton, New Zealand; 3MSD Animal Health, Global Ruminants, Boxmeer, The Netherlands.
*author for correspondence [email protected]
INTRODUCTION RESPONSE TO LAMB VACCINATION (EFFECT OF MDA)
MDA significantly (P < 0.005) decreased the immune response in lambs
In New Zealand (NZ) prevention of clostridial disease by vaccination of ewes 2-6 weeks pre- vaccinated with the reference vaccine. There was no association with MDA in
lamb normally protects lambs until weaning; however, in some flocks, Clostridium perfringens the test vaccinated lambs (Figures 1 and 2).
type D (i.e. Pulpy Kidney) deaths still occur. Additional vaccination at tailing (~3-8 weeks old) is The test vaccine was not only non-inferior, it was superior to, the reference
used to reduce this risk. As most lambs born to vaccinated dams will have maternally derived vaccine. (Figure 2).
antibodies (MDA), it is crucial that a vaccine is efficacious in the face of MDA. Few studies have The predicted probabilities of having an epsilon or tetanospasmin antitoxin
investigated the ability of clostridial vaccines to overcome MDA in sheep. At the time, the test concentration >5 or >2.5u/ml, respectively are shown in Figure 3. More lambs
product, Multine™ 5 (MSD Animal Health NZ, a vaccine containing toxoids of C. perfringens type were above these cutoffs in the test vs. the reference group for both epsilon
D, C. septicum, and C. tetani; and anacultures of C. novyi type B and C. chauvoei) did not have a [36% (17 to 55%)] and tetanospasmin [14% (3 to 24%) antitoxins.
label claim for use in lambs at tailing.
CONCLUSIONS
MATERIAL AND METHODS
• Lambs born to ewes vaccinated with test vaccine had higher MDA, than
To determine if the immune response in MDA positive lambs, vaccinated at tailing, with the lambs born to ewes vaccinated with reference vaccine.
test product was non-inferior to the immune response generated by the reference product
(Ultravac™5in1, Zoetis NZ Ltd). • MDA concentration significantly decreased the immune response in lambs
vaccinated with the reference vaccine.
MATERIAL AND METHODS
• The test vaccine overcame maternal antibodies and induced a protective
DESIGN immune response, superior to the registered reference product.
A single-site, randomised parallel group, non-inferiority field trial. The reference product was
chosen as it’s similar to the test product and the label indication is for use in lambs at tailing. • The test vaccine can be prescribed confidently for use in young lambs born
The study was conducted from Jul. 2012 to Jan. 2013 on a commercial farm in the (North Island) to ewes pre-lamb vaccinated with a clostridial vaccine.
of New Zealand.
SAMPLE SIZE & AEC (a)
Fifty lambs per group, assuming a non-inferiority margin 0.63, a SD 0.4u/ml, and alpha 0.05 and
beta of 0.2. Ethics Approval - No. 2/12. (b)
STUDY POPULATION
An mob of 126 maiden, twin-bearing, Romney, 2-tooth ewes, vaccinated with a two-dose
clostridial vaccination course, at most 12 months prior, and their surviving twin lambs (n=210).
They were managed as a single mob, until the weaning, and fed on ryegrass/clover pasture.
INTERVENTION
Prelamb, ewes were randomly allocated to the test (n=63), or reference (n=63) group and
given a single injection of the test or reference vaccine. The lambs from 10 dams randomly
chosen from each group remained as non-vaccinated controls. The remaining twins lambs were
randomly allocated, within pairs, to receive the test or reference vaccine. These lambs were
vaccinated at tailing (V1, median age 36 days, and at weaning 49 days later (V2). Blood was
sampled at V1 and V2 + 14 days. 19/20 control and a random selection of 50 twin lambs had
their sera analysed.
OUTCOMES
The response to lamb vaccination was measured by the epsilon (C. perfringens) and
tetanospasmin (C. tetani) antitoxin concentrations, EAT & TAT, respectively in lambs at V2
+14 days. Also the proportion of lambs with EAT or TAT concentrations above 5.0 or 2.5u/ml,
respectively and those that failed to respond, defined as a decrease in antitoxin concentration
from V1 to V2 + 14.
The response to ewe vaccination as measured by EAT and TAT MDA in lambs prior to V1.
STATISTICS
The geometric mean ratio (GMR) of the test, reference and control groups and the proportions
responding, were modelled using General linear models. Given the dependence between twins, dam
was included as a random effect. MDA was included as this was predicated a priori to be important.
Non-inferiority was met if the GMR 95%CI lower limit was above the non-inferiority margin of 0.63.
RESULTS
The baseline distribution of lamb gender and MDA (Table 1) were similar.
RESPONSE TO EWE VACCINATION (MDA)
Lambs born from test product vaccinated ewes had higher ETA and TAT MDA antibodies than
those from ewes vaccinated with reference vaccine. The GMR were 2.1 (1.7 to 2.7; P < 0.0005)
and 1.3 (1.0 to 1.7; P = 0.056), respectively.
TABLE 1
Distribution of epsilon and tetanospasmin antitoxin concentration (u/ml) in lambs at tailing (V1) by interventioin group.
Lamb Group n Min. Epsilon Max. Min. Tetanospasmin Max. FIGURE 1
19 Percentile Percentile
Control 50 0.8 25 50 75 9.7 0.3 11.5 Pairplots of epsilon (a) and tetanospasmin (b) antitoxin concentration in individual lambs prior
Reference 50 0.3 1.1 2.1 3.3 8.4 0.1 25 50 75 3.9 to vaccination at tailing (V1) and two weeks after a booster vaccination (V2 + 14) in control
0.3 1.0 2.6 3.7 10.5 0.1 1.1 2.0 3.0 4.5 (n= 19), reference (n = 50) and test groups (n = 50). The grey lines indicate an increased
Test 1.2 2.0 3.5 1.0 1.7 2.1 concentration, whereas the black lines show waning MDA and, in the vaccinated lambs, a
0.9 1.7 2.3 failure to response to vaccination.
(a) (b)
FIGURE 2 FIGURE 3
Non-inferiority plots of (a) epsilon and (b) tetanospasmin antitoxin concentration of test vs. reference product vaccinated lambs. Geometric Predicted probabilities and 95%CI of a lamb having an antitoxin concentration
mean ratio (hollow circle) and 95% CI (error bars). If the lower bound is to the right of the short-dashed vertical line this indicates non- greater than 5u/ml for epsilon, or 2.5u/ml for tetanospasmin.
inferiority. If it is to the right of the long-dashed vertical line (1.0) it indicates statistical superiority, i.e. P < 0.05.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
13 WBC 2016 (P02-002-135)
MSD ANIMAL HEALTH
Subcutaneous ischiorectal fossa injection: serological, colostral
immunity and tissue reactivity in rota, corona and E. coli F5 (K99)
vaccinated cows
John Moffat*1, Kate Golyn2; Richard Smith3, Jantijn Swinkels4
1MSD Animal Health; 2Vet Ent Otorohanga, 3Taranaki Vet Services, Patea, New Zealand New Zealand; 4MSD Animal Health, Global Ruminants, Boxmeer, The Netherlands.
*author for correspondence [email protected]
INTRODUCTION
Most labels for production animal products indicate vaccination in the anterior third of the neck. In New
Zealand’s (NZ) pasture-based production systems access to the neck of adult cattle on most farms is
impractical and, often dangerous. An alternative injection site at the rear of an animal that satisfies meat
quality and efficacy criteria is required. This study investigated the suitability of the ischiorectal fossa
(IRF - also called the caudal fossa, or tail notch). This area (YES in Figure 1) is the depression located
lateral to the tail head and medial to the tuber ischia. It is lateral to the caudal fold (NO in Figure 1), as
this reserved for TB testing in NZ.
OBJECTIVE FIGURE 1
To compare the immune response, measured by anti-rotaviral antibodies, and injection site reactivity, Injection site - subcutaneously in the ischiorectal fossa (Labeled YES). The
between cows vaccinated intramuscular (i.m.) in the anterior third of the neck (IM) with cows injected caudal fold is labeled NO.
subcutaneously (s.c.) in the ischiorectal fossa (IRF) using an oil emulsion vaccine containing inactivated
rotavirus, corona virus and E. coli F5 (K99); Rotavec Corona® (MSD Animal Health, NZ). FIGURE 2
MATERIALS & METHODS Boxplots of the natural log of rotaviral viral titres in colostrum by farm, breed
and injection site.
DESIGN
A multi-site, parallel-group, randomised non-inferiority field trial. The experimental unit was the individual
cow and the observational unit was the ith cow at each time point.
The study was conducted between July 2012 and October 2012 on two commercial, seasonally-calving
dairy farms in the North Island of New Zealand.
SAMPLE SIZE & AEC
A minimum of 60 cows per vaccinated group was determined by assuming a parallel-group design, a
non-inferiority margin of 0.63, a common standard deviation (SD) of 1.0 logu/ml., of 0.05 and 1−β of 0.8.
Animal Ethics Approval - MSD No. 2/12. α
STUDY POPULATION
123 healthy, multiparous, Friesian, Jersey and Friesian x cows, due to calve in the spring of 2012, that
had a history of vaccination with the test product in at least the previous calving season.
On each farm the trial cattle were managed as a single mob, and grazed on ryegrass/clover pasture until
they calved.
INTERVENTION
On each farm, cows were randomly assigned to either the IM or IRF group. Cows were injected once,
between about 3 and 10 weeks prior to calving. Rotaviral specific antibodies were quantified by virus
neutralisation (bovine rotavirus isolate, molecular typed as G6P5). Injection sites were scored on Days 0,
7, and 21.
OUTCOMES
The primary outcomes were rotaviral antibody titre in first colostrum and the degree, if any, of injection
site lesions.
STATISTICS
The geometric mean titre (GMT) of the test and reference and control groups, was estimated using
linear regression with treatment group as the predictor, conditioned on variables considered a priori to be
influential (i.e. farm, breed, age, and pre-vaccination serum titre). Model assumptions were evaluated by
examination of residuals and generation of Cook’s D values, and sensitivity analyses were conducted by
removal of potential influential observations. Non-inferiority was declared if the lower limit of the 1-sided
95%CI of the geometric mean ratio (GMR) of the GMT test and reference groups was above the non-
inferiority margin 0.63. This margin corresponds to a difference of about two thirds of a twofold dilution
in a serial dilution assay.
RESULTS FIGURE 3
113 (57 IM and 56 IRF) cows were included. Of the ten (6 IM. and 4 IRF) post-admission withdrawals, Mean (95%CI) of rotaviral titre in colostrum of cows vaccinated with Rotavec
most (7) were due to a failure to collect a colostrum sample. Corona either intramuscularly in the neck (i.m. neck) or subcutaneously in the
ischiorectal fossa (s.c. IRF).
Baseline age, breed and rotaviral serum antibody titre distributions (Table 1) were TABLE 2
similar across the treatment groups.
The distribution colostral rotaviral antibody titres were very similar on the IM and Summary of multiple regression model of rotaviral antibody titres in the colostrum of cows vaccinated
IRF groups. As shown in Table 2, colostrum from Jersey cows had on average ½ the pre-calving with Rotavec Corona (n = 113) as a function of injection site, farm, breed, age and day 0 titre.
rotaviral concentration of Friesian cows. Farm A cows typically had colostrum titres The GMT is the adjusted geometric mean and its associated 95% CI for each co-variate. The GMR is the
2.4 times that of cows on farm B. While breed and farm influenced the colostrum geometric mean ratio and its associated 90% CI.
concentration (Figure 2), there was no interaction with treatment (P>0.1, not shown).
In the final model (Table 2), injection in the IRF was non-inferior to traditional IM use. N GMT 95% CI GMR 95% CI P-value
Figure 3 shows the unadjusted means for each group; note, the unadjusted GMR was
0.97 and the lower bound was 0.68 for these data. LCL UCL LCL UCL
Covariate
No systemic adverse events were associated with vaccination. There was no evidence Injection site i.m. 57 1727 1274 2340 0.92 0.64 1.32 0.702
that the median volume was different between the different injection sites.
s.c. IRF 56 1587 1168 2157
CONCLUSIONS Farm A 58 1084 759 1547 2.40 1.47 3.90 0.003
Vaccination with an oil TABLE 1 B 55 2590 1792 3744
adjuvanted, inactivated
rotavirus, corona virus Distribution of rotaviral antibody titres in the serum of cows prior to Breed Friesian 92 1893 1489 2406 0.49 0.3 0.79 0.015
and E.coli F5 (K99) pre-calving vaccination.
subcutaneously in the IRF is Jersey 21 923 551 1544
as effective as intramuscularly Injection n Min. Percentile Max.
injection in the neck. This Site
gives NZ vets the confidence
to prescribe an alternative 25 50 75 Age 113 1656 1336 2053 1.07 0.97 1.17 0.234
injection site and hence
reduces the risk to farmers i.m. neck 57 50 282 565 1131 3200 Titre Day 0 1.09 0.89 1.32 0.493
of injury during routine Vaccination to calving 0.99 0.97 1.01 0.355
vaccination. s.c. IRF 56 100 350 583 966 3200 interval
MSD Animal Health 14
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P02-002-145)
MSD ANIMAL HEALTH
Does BVD vaccination of recently weaned pasture-fed beef cattle
prevent weight loss when exposed to persistently infected cattle?
John Moffat*1, Stu Bruere2, Jantijn Swinkels3
1MSD Animal Health, New Zealand; 2Vet Service Hawkes Bay, Masterton, New Zealand; 3MSD Animal Health, Global Ruminants, Boxmeer, The Netherlands.
*author for correspondence [email protected]
KEY FINDINGS CONCLUSIONS
WEIGHT GAIN & VACCINATION • BVDv rapidly spread through the study population.
• Transient Type I BVDv infection may affect growth in weaned pasture-fed cattle. • Vaccinated cattle had a markedly different immune response compared with
• Vaccination with Bovilis BVD may reduce the effect of infection (Figure 1).
• Marked amnestic response in the vaccinated cattle (Figure 2). non-vaccinated cattle.
• The greater ADG (56.1g/day) of the vaccinated cattle, suggests a protective
INTRODUCTION
vaccine effect; however the greater variability (i.e. SD of 134g/day vs. assumed
Bovine Viral Diarrhoea virus (BVDv) commonly infects New Zealand (NZ) cattle1 and 100g/day), meant the study had insufficient power to discriminate this
can cause significant production loss; especially reproductive failure2–4; however difference with 95% confidence.
little is known about ill-thrift associated with transient infection in young, weaned • This study indicates transient BVDv infection may affect growth in pasture-fed
beef cattle. cattle and vaccination may reduce this effect.
• Further studies are required.
OBJECTIVE
REFERENCES
To compare the growth rate of vaccinated vs. non- vaccinated weaned beef cattle
following exposure to persistently infected (PI) cattle. 1. Pérez MJ, Wilks CR, Rice M. Antibodies to bovine viral diarrhoea virus in beef
cattle. N Z Vet J 1994;42:73.
MATERIALS & METHODS
2. Perez MJ, Wilks CR, Vermunt JJ, West DM. Observations on BVD virus infection
DESIGN in New Zealand beef herds. N Z Vet J 1995;43:85–6.
• Single-site, randomised parallel group challenge field trial.
• Conducted between May and September 2014. 3. Voges H, Young S, Nash M, Improvement L. Direct adverse effects of persistent
• Wairarapa province (North Island) New Zealand. BVDv infection in dairy heifers – a retrospective case control study. VetScript
2006;4–6.
SAMPLE SIZE & AEC
• Minimum detectable difference in average daily gain (ADG) estimated as 63g/ 4. Heuer C, Healy A, Zerbini C. Economic effects of exposure to bovine viral
diarrhea virus on dairy herds in New Zealand. J Dairy Sci 2007;90:5428–38.
day, assuming a standard deviation (SD) of 100g/day, alpha 0.05 and beta 0.80.
• Animal Ethics Approval - MSD No. 31/12. Liveweight (kg) 300 PI Intro
V2
STUDY POPULATION 280
• 80 weaned, mixed-gender, spring-born (R1, August, 2013) Angus-x BVD Control Vaccinated
260
seronegative cattle. V1
• Managed as a single mob on brassica crop, with access to a run-off pasture,
240
meadow hay and ad lib water.
220 28 42 56 70 84 98
INTERVENTION 0 Days from first vaccination
• Randomised (blocked by weight and gender), into a non-vaccinated control or a
FIGURE 1
vaccinated group (n=40/group).
• Vaccinated with an inactivated Type I BVD vaccine (Bovilis© BVD, MSD Animal Mean weight of control (solid blue line, n=39) and vaccinated (red dashed line, n= 39) groups at
days 0 and 42, then every 14 days after the introduction of persistently infected BVD cattle (PI
Health, NZ) on days 0 (V1) and 28 (V2). Intro). - First (V1) and second (V2) vaccination with Bovilis BVD: Error bars SEM
CHALLENGE Geometric Mean Titre 2048 Vaccinated
• Six Persistently Infected (PI) (BVD Type I) calves Introduced 14 days after V2. Control
• Yarded together for 4 hours, and then run within the mob for the remainder of
1024
the study.
512 V1 PI Intro
OUTCOMES V2
• Viral neutralisation test (VNT) titre of BVDv specific antibody on days 0, 42, 56,
256 14 28 42 56 70 84
70, and 84. 128 Days from first vaccination
• Weight of calves on days 0, 42, 56, 70, 84, 98 and 112.
0
STATISTICS
• Average daily gain (ADG) between challenge and day 98 modelled using linear 0
regression -fixed effect of vaccination adjusted for baseline weight and gender FIGURE 2
(NB: data from day 112 was excluded due to a radical change in diet 24 hours
prior weighting). Geometric mean titre of control (solid blue line, n=39) and vaccinated (red dashed line, n=39)
groups at 14 days after vaccination (day 42), then every 14 days after the introduction of
RESULTS persistently infected BVD cattle (PI Intro). - First (V1) and second (V2) vaccination with Bovilis
BVD: Error bars 95%CI
BASELINE
• Weight and gender distribution similar - Control mean [SD] 224.8 [18.9]kg vs. TABLE 1
Treatment 224.1 [19.4]kg- Table 1. Baseline weight by gender and group
RESPONSE TO VACCINATION Female Male
• Fourteen days after second vaccination titres ranged 1:16 to 1:256 - median
n Weight (kg) n Weight (kg)
[inter-quartile range (IQR)] of 1:64 [1:32 to 1:128]
Mean SD Mean SD
EVIDENCE OF CHALLENGE
• All (39/39) non-vaccinated, cattle seroconverted within 14 days of PI Control 19 218.5 18.6 20 230.9 17.7
introduction. Vaccinated 17 217.2 17.8 22 229.5 19.2
• Four-fold increase in titre 38/39 vaccinated cattle within 42 days of PI
introduction.
IMMUNE RESPONSE
• Geometric mean titre (GMT) [95%CI] in the vaccinated cattle (1385, [167 to
266]) was 6.6 times greater ([4.7 to 9.2], P = <0.0005) than the non-vaccinated
cattle (211, [167 to 1751iu/]) 42 days after PI introduction (Figure 2).
WEIGHT GAIN
• Vaccination did not have an adverse event on growth. Both groups had similar
ADG from day 0 to prior to challenge i.e. Control, 980 [923 to 1038] g/day vs.
vaccinated, 1010 [953 to 1068]g/day; difference 30 [-51 to 110]g/day, P = 0.48).
• Vaccinated cattle, adjusted for gender and starting weigh, had on average a
56.1g/day greater ADG over the 56 days after the introduction of the PI cattle.
This difference tended towards statistical significance (95%CI -5.3 to 117.5g/day,
P= 0.073).
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
15 WBC 2016 (P02-002-146)
MSD ANIMAL HEALTH
Evaluation of quality and management of colostrum on
dairy farms in Spain
Carlos Carbonell1, Laura Ferreira1 and Laura Elvira1
1MSD Animal Health, Madrid, Spain
INTRODUCTION TABLE 1
The quality and management of colostrum is the most important risk Mean (± SD) IgG , fat, protein, nutrient concentration, somatic cell count and bacterial contamination of colostrum
factor for dairy calf health. Numerous studies have associated failure
of passive immunity transfer (FPT) to both short term effects, like N Mean Std. Deviation Minimum Maximum
increased neonatal morbidity and mortality of scours and BRD; and to
long term effects such as reduction in calf growth rate or decreases Colostrum IgG (mg/Ml) 74 37,2 11,7 13,0 65,0
in future milk productivity. However, to our knowledge, quality and
management of colostrum, has never been studied on Spanish dairy Fat (%) 77 5,4 3,3 1,0 16,0
farms.
Protein (%) 77 14,0 3,3 3,0 19,0
Lactose (%) 77 3,2 0,6 2,0 5,0
OBJECTIVES Lean solids (%) 77 19,6 3,4 9,0 26,0
To evaluate the quality of maternal colostrum fed to newborn dairy Total solids (%) 72 25,0 4,9 12,0 39,0
calves in Spanish dairy farms through: 1) proportion of colostrum that LOG SCC 81 5,8 0,8 3,8 7,2
meets the minimum IgG concentration (50 mg IgG/mL in colostrum) LOG 10 TPC (cfu/mL) 85 4,8 1,3 2,1 7,5
and bacterial contamination (TPC < 100.000 UFC/ml); 2) evaluate the
prevalence of FPT on dairy calves; and 3) determine the quantity of
colostrum recommended to reach the necessary IgG absorption (100 g IgG in colostrum).
MATERIAL AND METHODS Figure 1: IgG colostrum (% Brix)<16
17-18
Twenty dairy farms located in different regions of Spain were selected. In each farm a Figure 2: IgG calves serum (% Brix)19-20
three step program was implemented: 21-22
FIGURE 1 AND 2 23-24
A.Epidemiological questionnaire 25-26
IgG measured as % Brix in colostrum (Figure 1) and calves serum samples (Figure 2, (n=74, 2227-28
B.Sampling colostrum and calves sera and 8.4 Brix% cut-off, respectively) 29-30
>30
C.Recommendations to improve colostrum management.
3.9-4.4 5-5.4
To evaluate the colostrum fed to the newborn calves, two samples were collected just 110.11.8961...507.51----85659557------11--11>980968061177............14994492949494
before feeding, 1) refrigerated with azidiol for nutrient, somatic cell count (SCC) and
bacterial analysis (Foss MilkoScan , Foss Fossomatic, Total Plate Count (TPC) and Petrifilm
Coliform Count Plate (CCP); and 2) frozen for immunological quality (ELISA IgG , Bio-X
Diagnostics) and indirectly by % Brix (Digital Refractometer Pal 1, Atago). Additionally,
serum from calves 1-5 days old fed sampled colostrum was analyzed to determine IgG
and % Brix (as for colostrum). Statistical Analysis. Descriptive analysis of data (SPSS
15.0).
RESULTS
1) Colostrum quality. High quality colostrum has an IgG concentration >50 mg/mL
(McGuirk et al., 2010), or >22 % Brix (Bielmann et al., 2010). In our study, only 69.8%
of colostrum samples reached this concentration (see figure 1 and table 1). Moreover,
experts recommend < 100,000 cfu/mL total plate (bacteria) count (TPC). In our study,
only 32.4% of colostrum samples meet both criteria.
High TPC, CCP and SCC emphasize the importance of minimizing colostrum bacterial
contamination by properly milking, handling and storage of colostrum; and improving
mastitis control on farm.
2) FPT on dairy calves. At 1 to 5d of age, adequate serum IgG level is considered if >10
mg/ml (Godden, 2008), or >8.4% Brix (Deelen et al., 2014). In our study, 46/72 (64%)
calves reach it; however 36% do not and suffer from FPT (see figure 2 and table 1).
3) Quantity of colostrum. Successful passive transfer in Holstein calves requires at least
100 g IgG in the first colostrum feeding. To reach this quantity, what volume of colostrum
should producers feed? (table 3)
TABLE 2 TABLE 3
Mean ± SD, minimum and maximum IgG concentration in 1-5 day old calves serum Quantity of colostrum needed for successful passive transfer of immunity (IgG)
N Mean Std. Deviation Minimum Maximum Colostrum quantity (L) % Calves fed <100g IgG (High risk FTP)
30,00
Serum IgG (mg/ml) 72 12,09 5,81 2,00 17,00 2 83,8
Serum BRIX (%) 69 9,29 2,13 4,00 3 38
4 14
CONCLUSIONS
This study indicates that, on today’s Spanish dairy farms, the majority of calves are at risk for both FPT and pathogen exposure. Only 32.4% percent of colostrum
samples meet minimum quality requirements of IgG and TPC and 36% of calves suffered FTP.
Colostrum management should be improved on Spanish dairy farms
REFERENCES
√ McGuirk SM and Collins M (2004) Managing the production, storage and delivery of colostrum. Vet Clin North Am Food Anim Pract 20: 593–603
√ Gooden S (2008) Colostrum Management for Dairy Calves 24: 19-39
√ Bielmann et al., (2010) An evaluation of Brix refractometry instruments for measurement of colostrum quality in dairy cattle J Dairy Sci 93: 3713-3721
√ Deelen et al., (2014) Evaluation of a Brix refractometer to estimate serum immunoglobulin G concentration in neonatal dairy calves. J Dairy Sci 97: 1-7.
MSD Animal Health 16
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P02-002-106)
MSD ANIMAL HEALTH
Education of the Dairy Employee on Animal Care –
A Practical Approach using Dairy CARE 365
Michael W. Bolton1, Paula Ospina2, Elizabeth A. Cox1, J. K. Shearer3, Ben Bartlett4, Fred J. Muller5, Gregory J. Crosley6, Scott Nordstrom1
1Merck Animal Health, Desoto, 2Veterinary Extension Associate, QMPS, Ithaca, 3Iowa State University, Ames, 4Michigan State University, East Lansing, 5Cattle Strategies, PLLC, Sunnyside,
6Countryside Vet Clinic, Somerset, Michigan, United States
INTRODUCTION Illustrates both the geographic breadth of
program and number of milk processors
The increase in the size of dairy operations in the US in the past 2 decades involved
has led to the change of the employee constituent. Employees have changed
from family based and local hires to employees that originate from multiple 4) More than 2,500 Modules have been distributed with nearly a thousand
countries with cultural differences and language barriers. Another change has registrants to the Dairy CARE 365 website. (www.Dairycare365.com).
been the retailers desire for more transparency on the dairy. Due both to dairies’
industrial size and increase pressure from consumers (von Keyserlingk et al., 5) This program reconnects veterinarian with farm as they aid in development
2013) the retailers want assurance that proper operating procedures are in place of these necessary SOP’s.
to document that animals, producing food under their brands, are being properly
cared for. 6) Documented decrease in worker injuries due to animal interaction in a very
large dairy operation where Dairy CARE has been utilized as part of their
OBJECTIVES safety program.
Three objectives of the Dairy CARE 365 program are: 7) National Milk Producers Federation’s FARM
program has officially partnered with the
1) The primary focus of the program is to help educate the dairy employee in Dairy CARE program.
the areas of cattle handling, calf care, and care of special needs cows. This
includes addressing the area of proper euthanasia, and management of non-
ambulatory animals.
2) To illustrate to the consumer that the dairy community is concerned with
the welfare of the animals entrusted in their care with the veterinarian at the
center of the program.
3) To have a positive impact on the welfare of both the employee and the dairy
cow and her calf.
National Milk Producers Federation
National (FARM) program has partnered
with Dairy CARE 365
MATERIALS AND METHODS CONCLUSIONS
Two distinct set of materials constitute the educational content of the Dairy Everyone involved in the dairy community has the responsibility to aid in the
CARE initiative. improvement of both the quality of the employee’s work environment as
well as the welfare of the dairy animal. Dairy CARE 365 addresses both of
1) The core of program is a series of 7 training Modules (e.g. Stockmanship, these needs, and we have had many testimonials regarding increased worker
Handling Non-ambulatory Cow, Calf Care). retention and documentation of decreased worker injuries.
a) Self advancing slides or video (20 minutes in length). The milk processors have adopted this program to illustrate to the retailers
that there is documentation of employee training in the area of animal care.
b) Choice of English and Spanish when loading. Certainly the penetration of this program, if nothing more, illustrates the thirst
that the dairy community has for employee education.
c) Narrated assessment at conclusion (if literacy is a concern).
Further research is needed in the areas of improving delivery of adult
d) Certificate printed upon successful completion. education materials, retention of knowledge, and survey methodology to
assess their impact.
2) Second part of program is the Workshop and Standard Operating Procedures
(SOP’s). ACKNOWLEDGEMENTS
a) SOP’s are in WORD format and customizable for each dairy operation (also Rick Jackson – Dairy Marketing – Merck Animal Health
in English and Spanish). For enhancing relationships with the processors and organizing
many of the workshops.
b) Workshops are 3 hours in length and the dairies usually have, by the
conclusion of the meeting, created an Animal Care Commitment, discussed
2 SOP’s (Non-ambulatory cow and Euthanasia), and have also viewed a
training Module.
c) Each farm is given a booklet and a USB jump drive to aid in developing
SOP’s that were not covered at workshop, and these all complement
the Farmers Assuring Responsible Management (FARM) program widely
adopted by US dairies.
RESULTS
This program has been embraced by both the dairy community, milk processors
and the veterinary profession;
1) Delivered 60 Workshops across 23 States and Canada for more than fifteen
Milk processors, several veterinary schools, and many large veterinary
practices.
2) The processors are utilizing this program to assure the retailers that there is
increasing activity in the area of improving animal care.
3) There have been 1,500 farms at these workshops representing 2 million cows
and ten thousand dairy employees.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
17 WBC 2016 (P02-002-034)
MSD ANIMAL HEALTH
Vaccination of dairy cows in late gestation: effect on serum and
colostrum antibody titers for BRSV and Mannheimia haemolytica
Laurent NOUVELLON1, Vincent TESSIER2, Evelyne MICHEL3, Sébastien ASSIE4, Bertrand RIDREMONT2, Jantijn SWINKELS5
1GIE Synervet, Saint-Aubin du Cormier, France, 2MSD Santé Animale, Beaucouzé, France, 3LABOCEA 35, Fougères, France, 4ONIRIS, Lunam Université, Nantes, France, 5MSD Animal Health, Madison, USA
OBJECTIVES Number of calves
Vaccination of calves against BRD is overall effective but, in some field situations, i.e. high infectious 100
pressure, early infection in very young calves before active vaccination, this could not be enough to 80
protect them against disease.
60
The objective of the study was to observe whether vaccination of cows during gestation with a
respiratory vaccine influences levels of serum and colostrum antibodies in the cow and in their calves 40
after colostrum intake.
20
MATERIALS AND METHODS
BRSV antibody
The fieldwork was conducted by local veterinarians during the fall and winter 2013-2014 in Brittany. titers in calves’
Two hundred Holstein cows from 13 farms were randomly selected to either vaccination (n=100) with
antigens of BRSV, PI3 and Mannheimia haemolytica (Bovilis® Bovipast, MSD Animal Health) or no 4 8 16 32 64 128 256 512 serum
vaccination (n=100). Cows were vaccinated according to label; the first dose was injected between 8-6
weeks before expected calving and the second dose between 4-2 weeks before expected calving. All cows by: Sero-neutralizing antibody titers of BRSV, as measured
by a VN test, of all calves born from all cows (n=189)
Levels of antibody titers were compared between cows before the vaccination schedule and after
the calving in each group and between calves 2 days after their birth from both groups raised on the Number of calves
same farms. Seroneutralizing antibody levels of BRSV were measured by a VN test (LABOCEA 35). To
determine M. haemolytica antibody levels, a validated quantitative ELISA (MSD Animal Health) was used. 100
80
RESULTS
60
189 mother-calf pairs were included in the analysis. One hundred mothers were vaccinated between
8 and 2 weeks before calving and 89 were not vaccinated. The animals were followed in 13 different 40
herds and in each herd the number of vaccinated and unvaccinated animals ranged from 6-27.
20
The levels of BRSV and Mannheimia haemolytica antibodies measured at the first blood test before
vaccination did not differ between vaccinated and non vaccinated cows (p>0.05). BRSV antibody
titers in calves’
Following vaccination, at 2 days after calving, specific neutralizing antibody titers of BRSV tended
(p=0.07) to be higher in colostrum from vaccinated mothers compared to control cows. However, 4 8 16 32 64 128 256 512 serum
these levels were significantly higher (p=0.01) in the serum of calves born from vaccinated mothers
compared to those from non vaccinated mothers (Figure 1). Non-vaccinated cows by: Sero-neutralizing antibody titers of BRSV, as measured
by a VN test, of calves born from non-vaccinated cows (n=89)
After vaccination, significantly higher levels of M. haemolytica antibodies (p<0.001) were measured in
serum of cows, in the colostrum of vaccinated dams and in the serum of calves born from vaccinated Number of calves
cows (Figure 2), always in comparison to the control group.
100
M.haemolytica 80
antibody titers in
calves’ serum
14
12 60
10 40
8 20
6
BRSV antibody
titers in calves’
4 8 16 32 64 128 256 512 serum
Vaccinated cows by: Sero-neutralizing antibody titers of
BRSV, as measured by a VN test, of calves born from vaccinated cows (n=100)
FIGURE 1
Number of calves from all cows, non vaccinated cows and vaccinated cows
according to their VN titers against BRSV.
ELISA antibody titers ELISA antibody titers of ELISA antibody titers of
of M.haemolytica of all M.haemolytica of calves M.haemolytica of calves
calves born from all cows born from non-vaccinated born from vaccinated cows
(n=189) cows (n=89) (n=100)
FIGURE 2
Levels of serum antibodies against M. haemolytica for all calves born from all cows (n=189), calves born
from non-vaccinated cows (n=89) and calves born from vaccinated cows (n=100) 2 days after calving.
CONCLUSIONS (Belknap and al, 1991; Makoschey and al, 2012). A previously published field
trial performed in French farms (Assié and al, 2013) showed a tendency to a
This field study confirmed that vaccination of cows in late gestation, with a positive effect of vaccinating dry cows against BRD, using the same vaccine, in
commercial trivalent vaccine, increased the levels of serum and colostrum addition to vaccination of calves older than 2 weeks. Further research is needed
antibodies against 2 major respiratory pathogens. to confirm the impact of vaccinating pregnant cows in addition to vaccination
of young calves in herds with a high prevalence of BRD in calves less than 3
Some BRSV and Mannheimia haemolytica, challenge studies concluded that weeks old.
colostrum enriched with specific antibodies from vaccination with Bovilis®
Bovipast show partial protection against infection of these 2 pathogens 18
MSD Animal Health
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WBC 2016 (P02-002-153)
MSD ANIMAL HEALTH
The Healthy Calf – A Nationwide Survey on the IgG-Concentration
in Newborn Calves serum
Elke Rauch1, Nicola Franz2, Sven Reese3, Michael Erhard1
1Department of Veterinary Sciences, Chair of Animal Welfare, Ethology, Animal Hygiene and Animal Husbandry, Faculty of Veterinary Medicine Ludwig-Maximilians-University Munich, Germany,
2MSD Animal Health Germany, Intervet Deutschland GmbH, Unterschleißheim, Germany, 3Department of Veterinary Sciences, Chair of Anatomy, Histology and Embryology, Faculty of Veterinary Medicine,
Ludwig-Maximilians-University Munich, Germany.
OBJECTIVES TABLE 1
Worldwide, the average mortality of calves Mean (± SD), median and minimum IgG concentration in 3 different IgG concentration categories in newborn calves serum. An IgG concentration > 10.0 mg/
during the first weeks of life is still 10 to ml is considered as a successful passive transfer of IgG. Lower concentrations (< 10 mg/ml) are considered as a failure of passive IgG transfer (FPT). If the
15%. More than 80% of calf losses can be IgG concentration is > 5 mg/ml and < 10 mg/ml it is called partial failure of passive transfer (pFTP).
traced back to farm-specific management
problems. Improved colostrum management IgG groups N Mean Standard deviation (SD) Median Minimum
may help to reduce these calf losses. The 284 2.8697 1.51308 3.1 0
objective of this study was to evaluate Failure of Passive Transfer
colostrum quality by measuring the (< 5 mg/ml) 450 7.4154 1.39599 7.4 5
immunoglobulin G (IgG) quantity in serum of
newborn calves in Germany. Partial Failure
of Passive Transfer
MATERIAL AND METHODS
(5-9.9 mg/ml)
Using a questionnaire sent by mail to
farmers, farm related data were collected IgG > 10.0 mg/ml 508 14.5177 3.96566 13.55 10
such as the number of animals, type of
housing, diseases, especially diarrhea, and Total 1242 9.2809 5.43567 8.5 0
specific calf-related data such as breed,
parity of the dam, calving difficulties, age TABLE 2
of calf at blood withdrawal, time spent with
the dam and the time after birth, quantity Mean (± SD), median, minimum and maximum IgG concentrations (mg/mL) in serum of newborn calves according to the lactation number of their dams.
and method in which the colostrum was
fed. Additionally, colostrum monitoring data Lactation Number of Mean Standard deviation (SD) Median Minimum Maximum
of 296 farms located all over the country number calves
from February-September 2015 were
analyzed for IgG quantity in serum using a 1 101 8.6902 5.3763 8.4 0.5 32.82
Sandwich ELISA (Erhard et al., 1995). IgG
content of 1242 blood samples, 1 sample 2 256 8.3906 5.07576 7.565 0 31.14
from each calf, taken between 36 to 96
hours after birth, was determined. 3 241 9.2279 5.45255 8.9 0 24.5
To trace the health status of the examined 4 141 9.7962 5.43294 8.7 0.1 27.4
calves within the first eight weeks of
life, the farmers were asked to complete 5 64 10.8295 5.67208 10.7 0.2 28.7
another questionnaire on the occurrence
and time of onset of diarrhea and 6 40 10.8553 5.4546 9.85 4.2 28.5
pneumonia.
7 18 11.4572 6.16573 10.45 0.31 21.7
8 5 9.872 7.92893 7.06 4.5 23.9
9 2 12.65 2.19203 12.65 11.1 14.2
RESULTS 10 2 12.6 8.6267 12.6 6.5 18.7
The results show that nearly 23% of the Total 870 9.2693 5.42259 8.5 0 32.82
calves were completely (failure of passive
transfer, FPT, IgG-concentrations < 5 mg/ TABLE 3
ml serum, n= 284) and 36% partly (partial
failure of passive transfer, pFPT, IgG- Mean (±SD), median, minimum and maximum serum IgG- concentration (mg/ml) of newborn calves according to the method of application (bucket, drench or
concentrations 5-9.9 mg/ml serum, n= bottle) of their first colostrum.
450) undersupplied with IgG. Only 41%
of the tested calves showed sufficient Colostrum N Mean Standard deviation (SD) Median Minimum Maximum
IgG-concentrations (> 10 mg/ml, n= 508) application
(Table 1).
106 11.2575 6.01386 10.745 0 26.3
Parity of the dam affected serum IgG
levels of the newborns (p<0.001, Pearson Bucket 581 9.0273 5.23122 8.4 0 32.82
correlation R = 0.149) slightly positive
(Table 2).
The method by which colostrum was Drench 95 9.7239 5.31181 8.5 1.95 29.1
fed did not significantly influence calves’ Bottle 427 9.0767 5.42203 8.3 0 28.7
serum IgG concentrations. The median
IgG-levels ranged from 8.3 mg/ml (bottle),
over 8.4 mg/ml (bucket) to 8.5 mg/ml in the
drenched calves (Table 3).
The amount of colostrum fed during the CONCLUSIONS
first feeding had a significant influence
(p<0.001, Pearson correlation R = 0.144) on the serum IgG concentrations of There are still needs for clarification and action concerning the management
the calves. of calf rearing in Germany and most likely in other countries as well.
There was no significant difference in serum IgG concentration (p = 0.182)
between the calves that got diarrhea and those not getting diarrhea in the
first, second or third week of life. However, calves with diarrhea showed
higher IgG-concentrations (Median with diarrhea: 10.05 mg/ml; without
diarrhea: 8.05 mg/ml). Calves with pneumonia had also higher IgG-levels
(Median with Pneumonia: 11.10 mg/ml; without Pneumonia: 8.65 mg/ml,
p = 0.490).
MSD Animal Health
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19 WBC 2016 (P02-002-051)
MSD ANIMAL HEALTH
Protocols combining GnRH and a prostaglandin analogue
for the treatment of ovarian cysts in dairy cows
Ana Cristina Figueiredo1, Carlos Fernandes1,2, Joao Henrique Viana3, Mauricio R. Vieira4, Denis A. Antonio5, Henderson Ayres5, Joao Paulo Barbuio5
1Animal Reproduction, Biotran LTDA, 2Animal Reproduction, University of Alfenas, Alfenas, 3Animal Reproduction, Embrapa - Dairy Catlle, Juiz de Fora,
4Animal Reproduction, Consulte Consultoria, Governador Valadares, 5MSD Saúde Animal, São Paulo, Brazil
INTRODUCTION
Treatment of follicular and luteal (or luteinized) cysts with
prostaglandin F2alpha and GnRH may result in cure and
thus, improve fertility. The aims of this study were to
evaluate the effectiveness of a GnRH analogue (Fertagyl™
Saúde Animal – Brazil) combined once or twice or not at all
with a prostaglandin F2alpha analogue (Estrumate™ - MSD
Saúde Animal – Brazil).
MATERIAL AND METHODS Follicular Cysts Corpus Luteum Luteal Cysts Corpus Luteum
Before Treatment After Treatment Before Treatment After Treatment
During 8 months, all Holstein cows (N=1,100), on 4 dairy
farms in the Minas Gerais State, Brazil, were evaluated by FIGURE 1
ultrasonography (Mindray M5, Mindray) at 3-4 weeks after
calving to detect ovarian cysts. The cysts with no apparent Ultrasonography images before and 7 days after treatment of ovarian cysts in dairy cows. The presence of a corpus luteum at 7 days after
wall by ultrasound were classified as follicular, while treatment is considered as a cure.
those with the presence of a thick wall (>2mm) with the
characteristic echotexture of luteal tissue were classified as Ovarian cyst cure rate of the 5 different treatment protocols
luteinized. The treatments were pre-determined, regardless
if the cyst was follicular or luteal.
The treatment was considered effective if at 7 days after
treatment, the cyst had disappeared and instead, a corpus
luteum was present and / or if the intervals treatment-1st
service and treatment –conception were shortened.
RESULTS
CURE RATE
There was no farm effect on the % of cure of ovarian
cysts (P>0.05). Ovarian cysts were detected in 12.4%
(136/1,100) of the cows. 23.5% (32/136) of the cysts
were classified as luteinized. According to the literature Cure (%)
the proportion of luteal cysts can be higher. It is possible
that some luteal cysts have a wall thickness below the
ultrasound detection level and were classified as follicular.
Statistical analysis of treatment efficacy of all 5 groups was
performed by Chi-square method and is shown in Figure 2.
Adding Cloprostenol treatment to Gonadorelin treatment at
diagnosis (Group 4 and 5) resulted in a higher effectiveness
than treatment groups G1, G2 and G3 (P<0.05), probably
because of its effects in luteinized cysts. Treatment Protocol
FERTILITY PARAMETERS FIGURE 2
The interval treatment - first service and the interval
treatment - conception differed among treatments (P<0.05 Percentage of cure (%) measured by ultrasonography at 7 days after treatment of cows with follicular (n=104, blue bar) or luteinized (n=32,
Table 2). When a second Cloprostenol was included in the red bar) of ovarian cysts for the 5 different treatment groups (see Table 1). Ovarian cyst cure rate was significantly higher in treatment group
protocol (G5) there was a further reduction in the interval 4 and 5 compared to group 1, 2 or 3 (p<0.05).
treatment - conception, when compared
to treatment protocol G4. This reduction TABLE 1
occurred probably because the second
Cloprostenol injection promoted the lysis The treatment schedule of 5 different treatment protocols (G1, G2, G3, G4 and G5) used for treating cows (total n=136) with ovarian cysts at diagnosis (day 0,
D0) and at day 10 after diagnosis (day 10, D10) or no treatment (control).
of corpora lutea eventually formed after the Treatment Treatment
earlier Gonadorelin injection. Protocol # of cows
0,5 mg of Gonadorelin (D0) 0,53mg of Cloprostenol (D0) 0,53mg of Cloprostenol (D10)
CONCLUSIONS G1 (Control) 16 - - -
Ovarian cysts incidence in Brazilian Holstein G2 31 at diagnosis -
herds is high. Both cure rate and the interval G3 28 at diagnosis 10 days after
treatment – 1st service and the interval
treatment – conception can be improved
when Gonadorelin treatment at diagnosis G4 29 at diagnosis at diagnosis -
is combined with Cloprostenol. A second G5 32 at diagnosis at diagnosis 10 days after
cloprostenol application 10 days later may
further improve reproductive performance. Total 136
TABLE 2
Average interval treatment - 1st serviçe and interval treatment - conception with their standard deviation (SD) for the 5 different treatment groups.
Group (N) Interval Treatment – 1st Serviçe (days ± SD) Interval Treatment – Conception (days ± SD)
G1 16 61.22 ± 17.87a 71.87 ± 21.85a
G2 31 44.54 ± 16.44b 60.76 ± 19.38b
G3 28 30.87 ± 12.63c 48.34 ± 16.96c
G4 29 26.19 ± 14.25c 46.12 ± 15.61c
G5 32 18.33 ± 10.18d 35.07 ± 14.32d
a, b, c, d If letters differ per treatment group, intervals are significantly different (p<0.05).
MSD Animal Health 20
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P02-002-186)
MSD ANIMAL HEALTH
Equine chorionic gonadotropin treatment in a fixed-time artificial
insemination protocol in anestrus beef cows
Ana Cristina Silva de Figueiredo1,2; Carlos Antônio de Carvalho Fernandes1,2; Henderson Ayres3; João Paulo Barbuio3; Rafael Marin Chiummo3;
Denis Barbosa Alves Antonio3
1Animal Reproduction, University of Alfenas, 2Animal Reproduction, Biotran LTDA, Alfenas, 3MSD Animal Health, São Paulo – Brazil
INTRODUCTION Treatment day Treatment
About 11% of the 65.5 million Brazilian cows are artificially Control Group Gonadotropin group
inseminated. 70% of the cows have zebu genetics and are beef
breeds. The zebu females usually show a long postpartum anestrus D0 insertion of a 1g progesterone intravaginal implant
(>70 days) making conventional insemination ineffective during plus IM1 application of 2mg estradiol benzoate
this period. The purpose of this study was to determine whether
equine chorionic gonadotropin (eCG) treatment (Folligon®, MSD D+8 Removal of the implant and IM application of 0.5mg sodium cloprostenol
animal health) is effective in accelerating follicular growth rate, and in
improving ovulation rate in a fixed time insemination (FTAI) protocol 1,5 ml of saline (IM) 1,5 ml of gonadotropin (300 IU, IM)
in anestrus cows.
D+9 IM application 2mg of estradiol benzoate
MATERIAL AND METHODS
1intra-muscular
Forty anoestrus crossbred beef females (20 cows and 20 heifers),
with no corpus luteum and a ≥7mm diameter follicle, were TABLE 1
enrolled and submitted to the treatment protocol showed in table
1. Randomization was performed on day 8 (D8) according to the Fixed time artificial insemination treatment scheme of crossbred cows and heifers.
diameter of the largest follicle in the ovary.
D0 D8 D9
Ultrasound was used (Mindray-M5) with 7.5 Mhz linear transducer
for ovaries evaluation. Measurement of the largest follicle diameter Insert P4 device Removal P4 device Estradiol
was performed from D8 to D10, every 24 hours. From D11 to D12 plus estradiol plus Cloprostenol Plus Benzoate
the ovulation was observed by ultrasound, which was confirmed benzoate
on D17 (figure 1). The average of the largest follicle diameter was Folligon or Saline
compared daily between both treatment groups by the Tukey test.
The percentage of follicles that ovulated were compared between D0 D8 D9 D10 D11 D12 D17
treatment groups, using Fisher’s exact test. A P-value lower than
5% was considered significant. For statistical analysis, SAS software Ultrasound Evaluation
version 9.3 was used.
FIGURE 1
RESULTS
Scheme of treatment and ultrasonography evaluation.
Ultrasound images taken from D8 to D17 are shown in figure 3.
In both the cow and the heifer group separately and in all females Average follicular diameter (mm) 11.0 11.0
together there were no differences (P>0.05) in the average Saline 9.1
follicular diameter at D8, the day the animals were assigned to Gonadotrofin
their treatments. There was an increase in the rate of follicular
development in animals treated with eCG. In cows under this 10.3 10.0
treatment, the follicular development was faster, showing the tested
product activity (figure 2). 9.5 9.2
The percentage of cows that ovulated at the end of the protocol, 9.0
between D11 and D12, was higher in the eCG group than in the 8.8 8.9
saline (control) group (95.0 vs. 30.0% - P<0.05). The number of
females in the eCG group that had a corpus luteum on D17 was 8.0 D9 D10
higher than the saline (control) group (90.0 vs. 30.0% - P<0.05) D8 evaluation day
(figure 4).
FIGURE 2
Average of follicular diameter (mm) from D8 to D10 in cows treated with saline (control, n=20) or equine chorionic
gonadotropin (n=20) in a fixed time insemination program of Zebu beef cows in Brasil.
FIGURE 3 100.0 95.0 90.0
75.0
Ultrasonographic images showing the evolution of follicular development in both the equine Saline
chorionic gonadotropin and the control group. Gonadotrofin
CONCLUSIONS % of cows 50.0 30.0
It was concluded that equine chorionic gonadotropin treatment after 30.0
removal of a 1 gram progesterone intravaginal device accelerates 25.0
follicular development compared to the control group and improves
the ovulation rate in fixed time artificial insemination protocols in 0.0 Ovulation D11 Corpus luteum D17
anestrus Zebu beef cows.
FIGURE 4
The % of cows, in both the equine chorionic gonadotropin treated (n=20)
and in the saline treated (control, n=20) group, that showed ovulation on
day 11 and had a corpus luteum on D17.
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21 WBC 2016 (P02-002-182)
MSD ANIMAL HEALTH
One single dose of equine chorionic gonadotropin is sufficient for
superovulation of cows
Carlos Antônio de Carvalho Fernandes1,2; Ana Cristina Silva de Figueiredo1,2; Henderson Ayres3; João Paulo Barbuio3; Rafael Marin Chiummo3;
Denis Barbosa Alves Antonio3
1Animal Reproduction, University of Alfenas, 2Animal Reproduction, Biotran LTDA, Alfenas, 3Embrapa Gado de Leite - Brazil, 3MSD Saúde Animal, São Paulo – Brazil
INTRODUCTION Treatment
The main advantage of equine chorionic gonadotropin (eCG) is its extended half- Treatment day Control Group eCG group
life. We therefore hypothesized that only one injection is sufficient for induction
of superovulation. The objective was to verify the efficacy of one single dose of a D0 insertion of a progesterone implant
specific eCG product (Folligon® MSD Animal Health) for induction of superovulation of D+4 plus application of 2mg estradiol benzoate
bovines. D+8
D+9 10 ml of saline 10 ml eCG (2,000 IU)
MATERIAL AND METHODS
Removal of the implant and IM application of 0.5mg sodium cloprostenol
Twenty cycling crossbred females (10 cows and 10 heifers) were randomly divided (Estrumate® MSD Animal Health - Brazil).
in a crossover design and were assigned to 2 different treatment groups that were
repeated twice within a 40-day interval (table 1). IM application 0.1mg of Gonadorelin (Fertagyl® MSD Animal Health - Brazil)
Ultrasound (US) evaluations were carried out on D0 and from D4 to D9 on a daily basis TABLE 1
(figure 1). The number of >5mm ø follicles were counted. Another US evaluation was
made on D17 to determine the number of ovulations. The animals were uniformly Superovulation treatment scheme of crossbred cows and heifers.
divided according to the number of >5mm ø follicles on D4 - prior to first treatment.
D0 D4 D8 D9
RESULTS AND DISCUSSION Progestrone device Folligon or Removal P4 device Fertagyl
The ultrasound images of the evolution of follicular development during the trial is plus estradiol Saline plus Estrumate
shown in Figure 3. There was no heifer-cow category effect, therefore cows and benzoate
heifers data were analysed together. At D0, no differences were observed in number
of follicles and ovulations between both treatment groups (P> 0.05). The evolution D0 D0 D4 D5 D6 D7 D8 D9 D17
of the number of >5mm ø follicles of both treatment groups is shown in Figure 2. It
was observed that as early as 24 hours after application of eCG (D5), the number of Ultrasound Evaluations
follicles in the eCG treatment group was higher than in the control group (P <0.05) - a
difference that was maintained until D9. FIGURE 1
A gradual evolution of the average number of ≤5mm ø follicles between D4 and D9 Scheme of treatment and ultrasonographic evaluation
amongst animals from the eCG treatment group could be observed, which proves
that this treatment is capable of stimulating the development of new follicles for a 30.0
long period after application.
Average number of follicles >5mm Saline 24.1 24.0
As a result of the stimulation of eCG treatment at the beginning of the wave of Gonadotropin
follicular development, more follicles reached the end of the protocol (D9) presenting 22.5
hormonal conditions to ovulate, i.e., ovaries with a diameter of >9mm (Figure 4).
19.3
eCG Group Saline Group 15.0 14.5
Day Right Ovary Left Ovary Right Ovary Left Ovary 10.0
7.5 4.6
4.6 2.3 2.3 2.8 2.3
4
0.0 D4 D5 D6 D7 D8 D9
evaluation day
5 FIGURE 2
Mean number of follicles with a diameter > 5 mm in both ovaries of 20 female cows from day 4 (D4), the
day of treatment of saline or equine chorionic gonadotropin, until day 9 (D9).
16.0Average ovulation number
16.0
6
12.0
8.0
7
4.0
0.9
0.0 Saline eCG
Treatment group
8 FIGURE 4
Average number of ovulations in both ovaries on day 17 (D17) in 20 animals receiving either saline (n=10) or
equine chorionic gonadotropin (n=10).
9
CONCLUSION
The conclusion is that one single dose of 2000 IU of equine chorionic
gonadotropin is sufficient for effective promotion of follicular development
and super-ovulation of cows.
17
FIGURE 3 22
Ultrasonographic images showing the evolution of follicular development in the equine chorionic gonadotropin
treatment group and in the saline treatment group (control).
MSD Animal Health
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WBC 2016 (P02-002-197)
MSD ANIMAL HEALTH
Field prevalence of bovine respiratory pathogens by
PCR technique in France
Siddartha Torres1, Vincent Tessier2, Bertrand Ridremont2
1Merck Animal Health, Madison, United States, 2MSD Santé Animale, Beaucouzé, France
OBJECTIVE RESULTS AND DISCUSSION
Bovine Respiratory Diseases (BRD) are a major pathological complex in A majority of analysis showed the presence of bacteria with more than
cattle, with significant impact on farm economical results. The pathogen 80% of samples positive for Pasteurella multocida (up to 85% associated
agents responsible for these disorders are essentially viruses and bacteria, with another pathogen) and 46% of samples positive for Mannheimia
often associated on the field. Polymerase Chain Reaction (PCR) is a haemolytica. Haemophilus somni and Mycoplasma bovis were found to a
major technique of diagnosis, due to its better detection performances in lesser degree. Regarding viruses, Bovine Respiratory Syncytial Virus (BRSV)
comparison with traditional bacteriological methods. was detected in less than one third of all cases, half in association with M.
haemolytica.
MATERIALS AND METHODS
This prevalence study in French field farms confirmed the presence of the 2
Since December 2012, 2 major laboratories for diagnosis located major BRD pathogens :
respectively in Nièvre and Ile et Vilaine departments participate in a field
BRD epidemiological survey, with support from MSD Animal Health. The - M.haemolytica, for which one could think that it could be overestimated
sampled animals , from a few days to several months old, had generally because of the sampling technique (nasal swabs) and the testing method
clinical symptoms of respiratory disease in early evolution. After sampling (PCR). But former studies showed a correlation between nasal swabs and
by the vets either from deep nasal swabs (3 swabs per farm) (80% of all trans-tracheal aspiration on pooled samples (3 particularly).
samples) or trans-tracheal fluids (<10%) or pulmonary tissues of animals,
PCR was performed after mixing maximum 3 samples from the same BRD - BRSV, for which the prevalence of 30% could seem to be low. Is it
outbreak in the same farm. A total of 648 individual viral and bacterial because of the sampling technique in relation to the PCR testing ? Some
diagnosis were analyzed. studies set the transtracheal aspiration as the most sensitive sampling
method to isolate BRSV when PCR is performed. Or is it an effect of usual
BRSV vaccination in farms ?
FIGURE 1. Prevalence M. haemolytica and VRSB. % positive results
% positive results
FIGURE 2. Prevalence M. haemolytica (red colour) and VRSB (blue colour) by departement/area in France.
MSD Animal Health
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23 WBC 2016 (P03-003-101)
MSD ANIMAL HEALTH
Resumption of ovarian cyclicity and energy balance in
Holstein cows in France based on in-line milk measurements
Claire Saby-Chaban1, René Fournier2, Faouzi Lyazrhi1, Edouard Alix3, Jantijn Swinkels4, Sylvie Chastant-Maillard1
1UMR INRA/ENVT 1225 Host-pathogen interactions, Reproduction, Ecole Nationale Vétérinaire de Toulouse, France. [email protected], 2MSD Animal Health, Angers, France,
3DeLaval International, Tumba, Sweden, 4MSD Animal Health, Boxmeer, the Netherlands
INTRODUCTION
Resumption of postpartum ovarian cyclicity is one of the main
limiting factors of reproduction in Holstein cows since fertility is
negatively affected by the delay from calving to first ovulation. Most
available studies are based on experimental herds, performed in
specific conditions.
The purpose of this study was to describe current postpartum
cyclicity resumption in Holstein cows under farming conditions in
France. We focused on cyclicity resumption, cow energy balance and
their effects on reproduction.
MATERIAL AND METHODS
1247 Holstein cows •• Data recorded spring 2014-spring 2015
(21 herds) with DeLaval •• First 90 days of lactation
milking robots with Herd •• 38.3% of primiparous females
NavigatorND system
In-line measurements •• Progesterone (indicator of cyclicity) assayed FIGURE 1.
(progesterone, BHB) on average every 2.7 days from 20 to 90 days
postpartum (dpp) Examples of individual follow-up of progesterone (up), beta-hydroxybutyrate (BHB) and milk production (down). Herd
Determination of Navigator screenshots.
the onset of luteal •• Beta hydroxybutyrate (BHB; indicator of energy
activity (C-LA) and balance) assayed every 3 or 4 days during the first
21 dpp.
ketotic status
•• Calving to the onset of luteal activity (C-LA)
is defined as the number of days from calving to
the first day with milk progesterone concentration
higher than 5 ng/mL.
•• Ketotic status: If at least one milk BHB value was
greater than 0.15 mmol/L during the first 21 dpp,
the cow was declared affected by ketosis.
Artificial insemination •• Success of each insemination 50%
Pregnancy status 45%
40%
RESULTS % of cows 35%
• Nearly half the cows (45.3%; 565/1247) had resumed cyclicity 30%
before 30 dpp and 86.2% (1075/1247) before 60 dpp (figure 2). 25%
20%
• Cows with C-LA earlier than 60 dpp (912/1034) were inseminated
significantly earlier than those with later C-LA (85.0±26.5 days vs 15%
105.1±32.5 days; p<0.001). Similarly, their calving to conception 10%
interval was shorter (116±49.2 days vs 138±52.6 days; p<0.001)
(table 1). 5%
0%
• Overall incidence of ketosis was 19% (237/1247).
[20-30] [30-40] [40-50] [50-60] [60-70] [70-80] [80-90] [90]
• Cows with an early resumption of luteal activity (< 30 dpp) were C-LA (days)
those with the lowest incidence of ketosis (12.9% vs 24.0% for
cows with C-LA > 30 dpp). FIGURE 2.
• Cows with ketosis had a longer calving to first insemination Delay from calving to luteal activity (C-LA)
interval (86.2±27.7 days vs 92.7±28.6 days; p=0.005) and a longer
calving to conception interval (116.1±49.5 days vs 131.8±51.0 30
days; p=0.006).
25
CONCLUSIONS
cows with ketosis (%) 20
These data collected in commercial herds indicate an early cyclicity
resumption. Delay from calving to luteal activity is affected by 15
ketosis over the first two months of lactation and in turn, impacts
on fertility. This delay is thus of major economic importance. In-line 10
progesterone and BHB assays are very potent tools for dynamic
monitoring of reproductive and metabolic status. They especially 5
witness the quality of the transition period management: any
deviation of these indicators needs to be followed by appropriate 0 <30 30-60 >60
corrective management solutions. Further analysis of these data [N=565] [N=510] [N=172]
will focus on the patterns of postpartum cyclicity resumption C-LA (days)
(prevalence and association with fertility).
FIGURE 3.
Ketotic status according to the delay from calving to luteal activity (C-LA)
Time to C-LA (dpp) < 30 30-40 40-50 50-60 60-70 70-80 80-90 >90 P-value
2.4.10-16
Calving to first Nb cows 566 224 163 91 44 35 20 26 1.4.10-4
insemination interval (days) Mean±SD 81.9±25.6 83.5±27.1 84.5±22.8 92.0±27.5 98.5±29.6 99.5±23.5 99.2±23.1 128.0±42.4
Nb cows
Calving to conception Mean±SD 271 120 81 50 22 18 10 15
interval (days) 111.8±46.9 124.6±55.5 115.0±49.0 120.9±44.5 129.5±59.2 141.8±50.3 119.7±40.3 157.7±49.9
TABLE 1.
Reproductive performances according to the delay from calving to luteal activity (C-LA).
MSD Animal Health 24
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P03-003-213)
MSD ANIMAL HEALTH
Monitoring of BVDV by testing antibodies to NS3 protein in calves
vaccinated with an inactivated BVDV vaccine
H. Kuijk*1, G. Nijhoving1, L. van Duijn2, M. Mars2, W. Theuws3, H. Swam1, B. Makoschey1
1MSD Animal Health, Boxmeer, the Netherlands, 2GD Animal Health, Deventer, the Netherlands, 3DAC De Peelhorst, Uden, the Netherlands,
*correspondenting author: [email protected]
OBJECTIVES Vaccination Day 1 Day 28 Day 56 Day 112 Day 168
group (n=5) First vaccination Second vaccination VN ELISA VN ELISA VN ELISA
On Bovine Viral Diarrhea Virus (BVDV) free certified farms in the
Netherlands, cattle are vaccinated with the inactivated vaccine Bovilis ELISA ELISA
BVD (MSD Animal Health) to reduce the risk that new Persistently
Infected (PI) calves are born. The BVDV free status of a farm can be Control No vaccination No vaccination ELISA ELISA ELISA
monitored by 6-monthly spot blood testing of 5 calves in the age of group (n=5) ELISA ELISA
8 to 12 months for the presence of Non-Structural proteins (NS3)
antibodies(a,b). Sometimes calves have been vaccinated before they FIGURE 1.
have been tested. The aim of this study was to determine whether
vaccination with Bovilis BVD interferes with this monitoring system Schedule of serum tests and vaccinations per herd.
in calves.
FIGURE 2.
MATERIAL AND METHODS
Herd A: All 5 BVD vaccinated calves (9761, 9763, 9759, 9760, 9762) showed virus neutralizing (VN) titers, but didn’t
In 8 BVDV free certified herds, 5 calves in the age group 8-12 months develop NS3 specific antibodies as measured by ELISA.
were vaccinated with Bovilis BVD according to manufactures instructions
and 5 sentinel calves were left unvaccinated (figure 1). Blood samples FIGURE 3.
were taken from all 80 calves on day 1 (first vaccination), day 28
(second vaccination), day 56, day 112 and day 168 and tested for NS3 Herd G: All 5 BVD vaccinated calves showed virus neutralizing (VN) titers. Three vaccinated calves (1839, 1841, 1847)
antibodies in the Prionics BVDV ELISA. Blood samples taken on day showed no NS3 specific titers in ELISA and 2 vaccinated calves (1845 and 1898) showed low NS3 specific titers in
56, day 112 and day 168 were also tested for Virus Neutralising (VN) ELISA on day 56, these titers were undetectable on day 112.
antibodies (figure 1). In one herd (H) blood samples from all calves
between 1 and 16 months of age were tested for BVD virus.
RESULTS
All vaccinated calves showed high VN titers on days 56, 112 and 168.
In 5 herds (A,B,C,D,E) neither vaccinated nor unvaccinated calves
developed NS3 specific antibodies as measured by ELISA (figure 2).
In the herds F, G, H, all unvaccinated calves tested negative for
antibodies as measured by ELISA. In 1 herd (F) 1 vaccinated calf
showed a low titer in ELISA on day 56, but was tested negative at
day 112. In 1 herd (G) 2 vaccinated calves showed low titers in ELISA
on day 56, these titers were undetectable on day 112 (figure 3). In 1
herd (H) all 5 vaccinated calves showed moderate titers on day 56,
these titers declined to very low titers on days 112 and 168. Because
of the moderate titers on day 56, in this herd, blood samples from all
calves between 1 and 16 months of age were tested for BVD virus.
No PI animal was found.
In 6 out of the 8 herds included in this study, vaccination of the
calves did not at all interfere with the young stock monitoring using
NS3-ELISA. In 1 herd, it was required to test additional 5 calves by
antibody ELISA to confirm the BVDV-free status. It remains unclear
whether the relative high titers in the 8th herd were caused 1) by the
vaccination, 2) by transient BVDV infection, or 3) by infection from a
PI that was already removed from the herd at the time of testing.
CONCLUSIONS
Young stock BVD monitoring can be applied in herds vaccinated
with the inactivated BVDV vaccine used in this study. All vaccinated
calves developed high virus neutralizing antibody titers. Only a
small proportion of vaccinated animals developed low, short lasting
titers of BVDV-NS3 specific antibodies as measured by NS3-ELISA,
potentially hampering young stock BVD monitoring. It is therefore
recommended to either take blood samples before vaccinating or to
monitor on vaccinating farms by antigen detection methods.
REFERENCES
a: Makoschey, B. et al. Evaluation of the induction of NS3 specific
BVDV antibodies using a commercial inactivated BVDV vaccine in
immunization and challenge trials. Vaccine, 25, (2007), 6140-6145.
b: Kuijk, H. et al. Monitoring of BVDV in a vaccinated herd by testing
milk for antibodies to NS3 protein. The Veterinary Record (2008),
October 18, 482-484.
MSD Animal Health
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25 WBC 2016 (P03-003-029)
MSD ANIMAL HEALTH
Molecular epidemiology of bovine viral diarrhea virus in Spain,
spatial and temporal distribution.
F. Esperón1, L. Elvira2, A. Torre1, V. Delicado1, M. Álvarez3
1Animal Health Research Center, INIA-CISA, Valdeolmos, Spain; 2MSD Animal Health, Madrid, Spain; 3Animal Health Department, University of Leon, Spain
INTRODUCTION FIGURE 1.
Bovine viral diarrhea virus (BVDV) type 1 is by Maximum likelihood phylogram of the sequences obtained in this study; including references for all known BVDV-1 subtypes. BLACK circle
far the most frequent BVDV isolated in Europe sequences represent the cases genotyped. The group 1b is collapsed (including 68 sequences from Spain) for better visualization of the tree.
and Spain. Indeed, only recently BVDV-2 has been
reported for the first time with a very low prevalence CONCLUSIONS
in Spain: 99,3% BVDV-1 vs 0,66% BVDV-2 (3/452)
(Aduriz et al., 2015; Cerviño et al., 2015 and ADSGs The results obtained demonstrate the clear predominance of BVDV-1 subtype b in Spain since 1999 until
Galicia, 2015). nowadays.
Recent introduction of subtypes 1d and 1f highlights the importance of periodic genotyping to obtain relevant
OBJECTIVES epidemiological information about the evolution of the different subtypes along the time.
No BVDV-2 was found, confirming its very low frequency in Spain.
1) identify BVDV genotypes circulating in Spain; 2)
study spatial and temporal clustering, and 3) check REFERENCES
trends of diversity along the time.
• Aduriz et al. (2015) First detection of bovine viral diarrhoea virus type 2 in cattle in Spain Vet Rec Open 2,1.
MATERIAL AND METHODS • ADSGs Galicia (2015) Detectado genotipo 2 de BVD en una vaca de Galicia. Boletín de ANEMBE 109
• Vilcek et al. (1994) Pestiviruses isolated from pigs, cattle and sheep can be allocated into at least three
164 viremic sera collected 2007 to 2015 were
genotyped using a one-step RT-PCR which amplifies a genogroups using polymerase chain reaction and restriction endonucleaseanalysis. Arch Virol 136(3-4): 309-23.
288 bp of the 5’-UTR (Vilcek et al., 1994). Sequences
100% identical were grouped as nucleotide sequence
types (ntST). A maximum likelihood tree with a
bootstrapping of 1000 replicates was made with ntST
and the reference sequences (Figure 1). Additionally,
to investigate the spatial and temporal distribution,
neighbor-joining networks were constructed (Figures
2 and 3).
RESULTS
Main results revealed a high genetic diversity,
identifying 88 ntST out of 164 sequences obtained.
All of them genotype 1 (BVDV-1).
Among subtypes, 1b was the most frequent, 79.9%
of total sequences obtained (131/164) and 77.3% of
ntST (68/88). Other subtypes found were 1f, 1e, 1d
and 1h (7.9, 6.1, 4.3 and 1.8% total sequences and
11.4, 2.3, 6.8 and 2.3% ntST, respectively).
No regional or temporal clustering was observed,
with the exception of subtype 1h that was only found
in one region (Castilla and Leon) and one year (2014). In
fact, some frequent ntSTs were detected in up to six
different regions out of 10 included in the study.
Regarding the evolution of the diversity of subtypes, 1b
and 1e were found from 1999 to 2015 and from 1999
to 2013, respectively. Eight ntST were present since
1999-2002 to 2007-2015, revealing a certain stability of
predominant sequences. Subtypes 1d, 1f and 1h were
identified for the first time in 2007, 2008 and 2000,
respectively.
FIGURE 2. FIGURE 3.
Spatial clustering of BVDV sequences using a neighbor-joining network Temporal clustering of BVDV sequences using a neighbor-joining network
MSD Animal Health 26
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P03-003-090)
MSD ANIMAL HEALTH
Efficacy of 86μg Gonadorelin and Cloprostenol Sodium in a dairy
Fixed-Time Artificial Insemination program
Lina S. de Montigny†, Terry Katz†, Paul M. Baldwin‡, Jantijn Swinkels§
†Intervet Inc., d/b/a Merck Animal Health, Madison, New Jersey, United States, ‡MSD Animal Health Innovation S.A.S, Beaucouzé, France, §MSD Animal Health, Boxmeer, The Netherlands,
BACKGROUND FIGURE 1
Historically, the only registered indication The percentage of dairy cows diagnosed as pregnant on each study farm following an estrus synchronization
for an 86 μg dose of gonadorelin, as the program in which cows were injected with 86 µg gonadorelin acetate (treated) or saline (control) and 500 µg of
acetate salt (Fertagyl®) in the U.S. was for cloprostenol sodium in a fixed time artificial insemination program.
the treatment of ovarian follicular cysts in
dairy cattle. During 2013, two other GnRH Figures above columns are percentage of cows pregnant in that group at that site; figures in columns represent the
products gained approval in the U.S. for use number of cows in that group at that site.
in combination with a prostaglandin F2α to
allow single fixed-time artificial insemination Treatment Pregnancy Rate (%) P-value
(FTAI) in dairy cattle. The registered dose 0.0051
in the U.S. of 86 µg of gonadorelin is lower Control (saline) 17.8
than that of the other two GnRH products. N = 374 33.4
Consequently we set out to explore if this Treated
lower dose of gonadorelin would be efficacious in an Ovsynch FTAI
program for dairy cattle. (86 μg gonadorelin)
N = 370
OBJECTIVES
TABLE 1
The objective of this study was to demonstrate the efficacy of an
86 μg dose of gonadorelin, as the acetate salt, for use in sequence Pregnancy rates by treatment, and p-value.
with 500 μg of cloprostenol in an Ovsynch single FTAI program, for
dairy cattle. REFERENCES
MATERIAL AND METHODS 1. GonaBREED® FOI, 2013
http://www.fda.gov/downloads/AnimalVeterinary/Products/
A large clinical field study was conducted across 7 dairy farms in 6 ApprovedAnimalDrugProducts/FOIADrugSummaries/UCM338852.pdf
US states. Healthy non-pregnant cows, 51-119 days in milk, were
randomized within farms to a gonadorelin-treated group (2mL 86μg 2. Chenault, J. et al. 2014. J. Dairy Sci. 97:2816-2821.
gonadorelin) or a saline-treated (placebo) control group. Most cows
were first or second parity (gonadorelin group 62.1%; control group 3. Fertagyl® FOI, 2015
64.6%). Enrolled cows were treated on Day 0 with 2 mL Fertagyl® http://www.fda.gov/downloads/AnimalVeterinary/Products/
(86μg gonadorelin) (N = 377) or 2 mL saline (N = 381); on Day 7 ApprovedAnimalDrugProducts/FOIADrugSummaries/UCM446093.pdf
with 2 mL Estrumate® (500μg cloprostenol) and on Day 9 with 2 mL
Fertagyl® (86μg gonadorelin) or 2 mL saline. Cows were then bred by
single FTAI on Day 10, 16 ± 8 hours after the Day 9 injections.
Pregnancy checks were performed by ultra-sound scanning and/or
rectal palpation at least 30 days later.
Of 758 cows enrolled, 744 were included in the efficacy analysis.
Pregnancy rates were analyzed for treatment effects by a generalized
mixed model for binomials with random terms for site and
interactions, using a Kenward-Roger adjustment (SAS/STAT version
9.3). Treatment efficacy was judged by checking if the pregnancy rate
of the treated group was significantly higher compared to the placebo
control group.
The study was conducted according to FDA-CVM Guidance for
Industry #85, Good Clinical Practice (VICH GL9).
RESULTS
The pregnancy results achieved on each farm are shown in figure 1.
The overall pregnancy rate (PR) was significantly higher (p=0.0051)
in the gonadorelin group compared to the control group (33.4% vs.
17.8%). There were no treatment-related adverse events. The 33.4%
PR obtained with a 86 μg dose of gonadorelin compares favorably to
the 33.4% PR reported for a 100 μg dose of gonadorelin present as
the acetate salt (GonaBREED®) [1], and also with the 32.2% PR that
has been reported in an Ovsynch FTAI program using a 200 μg dose
of gonadorelin present as the hydrochloride salt (Factrel®) [2].
CONCLUSIONS
This study demonstrates that an 86 μg dose of gonadorelin (present
as the acetate salt) used in sequence with 500 μg cloprostenol is
effective for estrus synchronization of dairy cattle in FTAI programs.
The effectiveness of this lower dose of gonadorelin (86 μg) compares
favorably with that reported for higher doses of gonadorelin.
Registration of the use of 86 µg of gonadorelin in FTAI programs was
granted in the U.S. in April 2015[3].
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
27 WBC 2016 (P03-003-234)
MSD ANIMAL HEALTH
Effect of two different bovine somatotropins on mmiillkk pprroodduuccttiioonn
and reproduction of dairy cows in Brazil
Henderson Ayres11, MMaarrcciilliioo NNiicchhii22,, SSaannddrroo LL..VViieecchhnniieesskkii33,, PPiieettrroo SSaammppaaiiooBBaarruusseelllii22,,RRooddrrigigooAAlmlmeeididaa44
11MSD Animmaall HHeeaalltthh,, BSrãaoziPl; a2Duleop, aBrrtamziel;n2tDoefpAarntimeanl tRoefprAondiumcatiloRneopfroUdSuPc,tiSoãnooPfaUuSloP,,SSPã,oBPraazuil;o3,SStaPr,-BMrailkzilF; a3Srmta,r-CMéiulkAFzaurlm, P, RC,éBurAazziul;l,4UPnRiv, eBrrsaidzial;d4eUnFievdeersraidl addoePFaeradneára, lPdRo, PBararaznilá, PR, Brazil
1,20 Group 1 Treat: P < 0.01
OBJECTIVE MMAATTEERRIIAALLAAGNrNouDpD2 MMEETTHHOODDSS Day: P < 0.01
The objective of thhiiss ttrriiaall wwaass ttoo eevvaalluuaattee mmiillkk yyiieelldd aanndd aasssseesssstthheerreepprroodduuccttiivvee Treat x day: P < 0.01
outcomes of a higghh--pprroodduucciinngg ddaaiirryy ccaattttllee hheerrdd iinn rreessppoonnssee ttoo ttwwooccoommmmeerrcciiaall
sources of bovinee ssoommaattoottrrooppiinn ((bbSSTT)) aaddmmiinniisstteerreedd ssuubbccuuttaanneeoouussllyyaattaaddoosseeooff EEigighhtty1y,1HH5oolslstteeininccoowwssffrroommaaccoommmmeerrcciaial lffaarrmmloloccaatteeddininththeessoouuththrereggioionn
500 mg, 16 timess wwiitthh aa 1144 ddaayyss iinntteerrvvaall.. Relative milk yeild, kg/day
ooff BBrraazzili,l,aavveerraagginingg11..7766±±11.1.166lalaccttaattioionnss,,7777.1.1±±2211.9.9ddaayyssininmmiliklk(D(DIMIM) )aanndd
3377..55±±66..55kkgg//ddooffmmiliklkwweerreerraannddoommlylyaassssigignneeddttoooonneeooff22trtereaatmtmeennttggrorouuppss. .
GGrroouup1p,1110 wwaassttrreeaatteeddwwitithh550000mmggbbSSTT(B(Boooossttinin,,MMSSDDAAnnimimaal lHHeeaaltlhth))ththee
ootthheerrggrroouupp,,((ggrroouupp22))wwaassttrreeaatteeddwwitithh550000mmggooffbbSSTT(L(Laacctototrtoroppinin, ,EElalannccoo) )
1,05
MMIILLKK PPRROODDUUCCTTIIOONN 1,20 1,00 Group 1 (C)
1,20 Group 2
Group 1 Treat: P < 0.01
1,15 0,95 Group 2
50 Group 1 Treat: P = 0.79 0 Day: P < 0.01
50 Group 2 Day: P < 0.01 2 TTrreeaatt:x day: PP << 00..0011
45 Group 1 TTrreeaatt:x day: PP <= 00..0719 1,15 Day: P < 0.01
45 Group 2 Day: P < 0.01
40 Treat x day: P < 0.01 RelatRiveleatimvilek ymileilkdy,eiklgd/,dkagy/day1,10 Treat x day: P < 0.01 16
40 X 1.000 SCC, per mL of milk 1,10 4 6 8 10 12 14
35
AverAavgeeraMgilek yMileikldy,eiklgd/,dkagy/day 35 Day after administration
30
30 UUDDDDEERRHHEEAALLTTHH
25
25 1,05 800 Treat: P = 0.31
20 1,05 600
20 0 Group 1 Day: P = 0.13
1,00 Group 2
0 1,00 Treat x day: P = 0.58
(A) 0,95 2 4 6 8 10 12 14 16
(A) 0,95 0 400 4 Day6 after ad8ministra1t0ion 12 14 16
50 100 150 200 250 0 2 Day after administration
200 250
50 Days afte1r0a0d 1st Adm1in5i0stration
Days after ad 1st Administration 200
42 Group 1 Treat: P < 0.01 (D)
42 Group 2 Day: P < 0.01
Group 1 TTrreeaatt:x day: PP << 00..0011 0
40 Group 2 Day: P < 0.01 -2 0 2 4 6 8 10
40 Treat x day: P < 0.01 Week after 1st administration
AverAavgeeraMgilek yMileilkdy,eiklgd/,dkagy/day 38 FFIIGGUURREE 22
38
CCLLININICICAALLMMAASSTTITITISIS::OOffththee8800aannimimaalslsinin222244trtiraial lddaayyss, ,1177aannimimaalslshhaaddcclinlinicicaal lmmaasstititsis, ,oof fwwhhicihch88
36 ccaasseessininttrereaatmtmeennttggrorouupp11aanndd99ccaasseessinintrtereaatmtmeennttggrorouupp22. .
36
(B) TTAABBLLEE 11
34 (B)
34 AAvveerraaggeerreepprroodduuccttiviveeoouutctcoommeessooff8800ccoowwssininaahhigighhpprorodduucciningghheerdrdininreressppoonnsseetototrtereaatmtmeennt twwitihth22
2 4 6 8 10 12 14 16 ddififeferreenntt ccoommmmeerrcciaial lssoouurcrceessooff bboovvinineessoommaatototrtoroppinin(b(bSSTT).).
0 2 4 Day6 after ad8ministra1t0ion 12 14 16
0
Day after administration
TTrreeaattmmeennttPPrroottooccooll GGrroouupp11 GGrroouupp22 PP
1,20 Group 1 Treat: P < 0.01 DDIIMM 7744.9.9 7799.4.4 >>00.2.200
1,15 Group 2 PPaarriittyy 11.7.755 11.7.788 >>00.2.200
Day: P < 0.01 PPRRaatttthheeiinniittiiaall 22.5.5%% 1122.5.5%% >>00.2.200
Treat x day: P < 0.01
Relative milk yeild, kg/day 1,10 PPRRaatttthheeeenndd 5500.0.0%% 5555.0.0%% 00.3.322
1,05 ##ooffSSeerrvviicceess 33.4.455±±00.5.544 33.4.499±±00.4.400 00.9.933
IICCPP 112299.7.7±±2222.9.9 113300.4.4±±1177.1.1 00.9.977
1,00 (C)
DDIMIM––DDaayyssininMMiliklk;;PPRR––pprereggnnaannccyyraratete;;##––nnuummbbeersrs; ;ICICPP––inintetervrvaal l(d(daayyss) )bbeetwtweeeennccaalvlvininggaanndd
pprreeggnnaannccyy
0,95
0 2 4 6 8 10 12 14 16
Day after administration
FIGURE 1
AAvveerraaggee ddaaiillyy mmiillkk yyiieelldd ((AA)),, aavveerraaggee wweeeekkllyy mmiillkkyyiieelldd((BB)),,aavveerraaggeerreellaattiviveemmiliklkyyieieldld((CC))aannddaavveerraaggeessoommaatticiccceelll
ceoaoacuuchhnnttrrsseep(p(DDrree))ssooeeffnnaattiihnhnigigg8gh0thtr-r0-epepararototmdmduueeccnniintntgwgwdidittahahiiGrrayayrdocdciuaifafftpftetetlrle1reeenhnhtetercrcddoom(m(nnm=m=88ee00rrc)c)iiiainDTanllrarserseyeoaos:sutup:prrococnenessoeoefftbtobooPoP2v2vi==nitntrer0e0ee.a.sa13stot3m1ommmeeanantttotogtgtrrrorooopupuipnipnss((b(b(GSGSTrTro)o).u.upp11aannddGGrroouupp22))
Group 2 Treat x day: P = 0.58
600 X 1.000 SCC, per mL of milk
CONCLUSION
This study show40s0tthhaatt iinn ttrreeaattmmeenntt ggrroouupp 11,, mmiillkk pprroodduuccttiioonn wwaassssiiggnniifificcaannttllyy
higher than in treaattmmeenntt ggrroouupp 22.. NNoo ddiiffffeerreennccee iinn uuddddeerr hheeaalltthh,,nnoorriinn
reproduction outccoommeess wwaass ffoouunndd bbeettwweeeenn ttrreeaattmmeenntt ggrroouupp 11aanndd22..
200
(D)
MSD Animal Hea0lltthh-2 0 2 4 6 8 10
Copyyrriigghhtt ©©22001166 IInntteerrvveett IInntteerrnnaattiioonnaall BB..VV,, aallssoo kknnoowwnnaassMMSSDDAAnnimimaallHHeeaaltlthh..AAlllrrigighhttssrreesseerrvveedd..
Week after 1st administration
WBCC 22001166 ((PP0034--000034--211256) 28
MSD ANIMAL HEALTH
Comparison of Two Macrolides In the Control Of BRD
in Calf Ranch Cattle
Norman Stewart*1
1Technical Services, Merck Animal Health, De Soto, United States
*Presenting Author
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
29 WBC 2016 (TP1-041-001)
MSD ANIMAL HEALTH
Efficacy of flunixin meglumine pour-on administration in a
tissue cage model of inflammation in cattle
Olivier Roy*1, René Fournier2, Julien Thiry2, Philippe Brianceau3, Siddartha Torres3
1CEBIPHAR, Fondettes, France; 2MSD Santé Animale, Beaucouzé, France; 3Merck Animal Health, Madison, United States
*Presenting Author
MSD Animal Health 30
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (TP1-041-002)
MSD ANIMAL HEALTH
Early detection of bovine respiratory disease and immediate
treatment with flunixin help to reduce the use of antibiotics in
beef calves
C. C. Gelfert*1, C. Ströbel2, C. Beisl3, E. Thesing4, D. Lange1
1Intervet Deutschland GMBH, Unterschleissheim, 2Ströbel Christina, Beratungsdienst Rindermast, Ilshofen, 3Gassner Franz & Beisl Josef, Tierärztl. Gemeinschaftspraxis, Frontenhausen,
4Intervet Deutschland GMBH, Unterschliessheim, Germany
*Presenting Author
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
31 WBC 2016 (TP1-041-003)
MSD ANIMAL HEALTH
Treatment of naturally occurring bovine respiratory disease in
juvenile calves with a single administration of a combination of
florfenicol plus flunixin meglumine
Julien Thiry*1, Vincent De Haas1
1R&D, MSD Animal Health, Madison, NJ, USA
*Presenting Author
MSD Animal Health 32
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (TP2-056-001)
MSD ANIMAL HEALTH
Associations between lung consolidation, bovine respiratory
disease treatment and live-weight in UK veal cattle.
Williams, P.*1, Potter, T.2, Cooper, R.3
1MSD Animal Health, Walton Manor, Walton, Milton Keynes, MK7 7AJ, UK, 2Westpoint Farm Vets, Dawes Farm, Bognor Road, Warnham, West Sussex RH12 3SH, UK
3Evidence Based Veterinary Consultancy, Rural Enterprise Centre, Redhills, Penrith, CA11 0DT, UK
*Presenting Author
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
33 WBC 2016 (YS2-046-002)
MSD ANIMAL HEALTH
On farm monitoring of the four Qs of colostrum management is
important
Hart, K.R.*1, Reyher, K.K.2, Barrett, D.C.2, Williams, P.D.3
1George Farm Vets, Malmesbury, Wiltshire, SN16 9AU, 2School of Veterinary Sciences, University of Bristol, Langford House, Langford, Bristol, BS40 5DU,
3MSD Animal Health, Walton Manor, Walton, Milton Keynes, MK7 7AJ, UK
*Presenting Author
MSD Animal Health 34
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (YS2-046-003)
MSD ANIMAL HEALTH
Effectiveness of Tildipirosin and Tulathromycin in the control of
bovine respiratory disease in high risk beef heifers
Riccardo Compiani*1, Stefano Jottini2, Siddartha Torres*3
1Veterinarian practitioner, Milano, Italy; 2MSD Animal Health, Milano, Italy, 3Merck Animal Health, Madison, United States
*Presenting Author
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
35 WBC 2016 (TP2-056-005)
MSD ANIMAL HEALTH
Flunixin meglumine transdermal pour-on solution as adjunct
therapy in the treatment of bovine respiratory disease in calves
less than 8 weeks of age
Julien Thiry1, Siddartha Torres1, Vincent de Hass1, Philippe Brianceau1
1Merck Animal Health, Madison, United States
OBJECTIVES 41 Flunixin Carprofen
40.5
Flunixin has anti-inflammatory, anti-pyretic and analgesic effects. Rectal temperature (ºC) FIGURE 1
This molecule is commonly used for the relief of pain and control of 40
inflammation and pyrexia associated with diseases of different origin 39.5 6 hours post-treatment (Time 6h) Effect of treatment on
and nature. A novel 50 mg/ml flunixin transdermal formulation was mean rectal temperature
developed by MSD Animal Health (Finadyne® Transdermal) and is now 39 (°C) 6 hours after
the first NSAID registered to be administered as a pour-on product along 38.5 dosing. The difference
the dorsal midline in cattle. The objective of the present study was to is statistically significant
determine the field effectiveness of the 50 mg/ml flunixin transdermal 38 (p<0.0001).
formulation as adjunct therapy in the treatment of BRD in juvenile Enrolment (Time 0h)
calves.
100% 0 1234 0.00%
MATERIAL AND METHODS 90% 13.04% 0.00% 11.54%
80%
A total of 49 Holstein calves, originating from German auction markets, 70% 15.38% 21.74% 34.62%
less than 8 weeks of age, showing severe signs of respiratory disease
(depression score ≥2, respiratory score ≥2 and rectal temperature 60% 56.52% 76.92% 78.26% 53.85% FIGURE 2
≥40.3°C), were enrolled and and randomly assigned to treatment. 50% Carprofen
7.69% Flunixin Effect of treatment on mean
Each animal was treated either with the test product, 50 mg/ml flunixin 40% 0% respiratory characters score
(Finadyne® Transdermal; 3.3 mg/kg flunixin; 1ml/15kg; MSD Animal Carprofen 6 hours after dosing. All
Health) administered topically once along the dorsal midline, or the 30% animals were examined for
control product, 50 mg/ml carprofen (Rimadyl®; 1.4 mg/kg carprofen; 20% 30.43% the characterisation of the
1ml/35kg; Zoetis) administered by subcutaneous injection once, on day 10% respiratory signs such as
0. Animals from both treatment groups received cefquinome (Cobactan® 0% 0% tachypnoea (or polypnoea) (≥
LA 7.5%; 1ml/30kg cefquinome; 2.5 mg/kg; MSD Animal Health) 40 breaths/minute), dyspnoea
administered by subcutaneous injection on day 0 and day 2. To avoid Flunixin (abnormal respiration), cough
observation bias and to preserve masking, treatments were dispensed and mucopurulent nasal
and administered by individuals not involved in the clinical assessment. discharge. Each sign counted
for one point to establish
All animals were observed for clinical signs of disease for 6 hours respiratory character score.
following treatment initiation and daily thereafter for 5 consecutive days.
The dosing sites was evaluated and any adverse effects were recorded. Normal Mild Moderate Severe
All animals underwent nasopharyngeal swabbing for bacterial culture, 100% 0.00% 0.00%
identification and determination of cefquinome sensitivity. 90% 100.00%
80% 100.00% 100.00% FIGURE 3
RESULTS 70% 0% Flunixin Carprofen
60% 100.00% Effect of treatment on mean
Based on the prevalence of the isolated pathogens in the pre-treatment 50% Carprofen depression score 6 hours
samples, the outbreaks can be attributed to Mannheimia haemolytica 40% after dosing. All animals were
(46.9%) and Pasteurella multocida (46.9%). Mycoplasma bovis (16.3%) 30% examined for the depression
and Histophilus somni (4.1%) were also detected (Table 1). 20% such as normal; mild; moderate
and severe.
All bacteria, except Mycoplasma bovis and spp, isolated during the 10%
study (at pre-treatment and at treatment failure) were susceptible to 0% 0%
cefquinome.
Flunixin
Both treatments were effective to reduce pyrexia; however, the
decrease in temperature was significantly greater with flunixin TABLE 1
transdermal (-1.68°C) compared to the positive control group (-1.02°C)
(Fig. 1). This difference was statistically significant (p<0.0001). Summary of efficacy results Flunixin Carprofen
Variable 23 26
Flunixin transdermal acted quickly and was efficacious to prevent the N enrolled animals
deleterious effects of inflammation as it was shown by the clinical Age (days) at enrolment 42.52 ± 5.92 44.69 ± 7.13
index (depression and respiratory signs) improvement at 6 hours post- Body weight (kg) at enrolment 62.70 ± 8.44 66.79 ± 10.54
treatment (Fig. 2 and 3) and within each post-treatment day (data not Rectal temperature (°C) at enrolment 40.67 ± 0.29 40.62 ± 0.27
shown). Rectal temperature (°C) at 6 h after treatment 38.99 ± 0.42 39.60 ± 0.63
Drop in temperature 6 h after treatment a
A very low number of treated animals presented dosing site reactions Treatment success rates on day 5 b -1.68 °C -1.02 °C
throughout the observation period. 100% (23/23) 100% (26/26)
Neither flunixin nor control had a negative influence on the health status a The decrease in rectal temperature 6 h after treatment was statistically significantly greater in the
including appetite and faecal consistency. flunixin group compared to the carprofen group (p<0.0001).
CONCLUSIONS b The non-inferiority of the flunixin group compared to the carprofen group was statistically
demonstrated.
Prompt treatment of sick calves with the novel flunixin transdermal in
conjunction with an appropriate antibiotic helps reduce fever, control
bacterial replication, and reduce the likelihood of permanent lung
lesions. This study demonstrates that the single dermal application of
Finadyne® Transdermal 50 mg/ml pour-on solution is a safe and effective
therapy to aleviate the signs of inflammation associated with BRD.
REFERENCE
Thiry J., Brianceau P. Flunixin meglumine transdermal pour-on solution as
adjunct therapy in the treatment of bovine respiratory disease in calves
less than 8 weeks of age. Journal of Veterinary Science and Research
(2014) 1:007.
MSD Animal Health 36
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WBC 2016 (P04-004-114)
MSD ANIMAL HEALTH
Efficacy of injectable florfenicol plus flunixin combination
against an experimentally induced Mycoplasma bovis
infection in calves
Julien Thiry1, Sophie Rubion2, Cliff Ramage3, David Reddick3, Vincent de Haas1
1 Merck Animal Health, Madison, United States, 2MSD Animal Health Innovation, Beaucouzé, France, 3Moredun Scientific Ltd, Penicuik, United Kingdom
OBJECTIVES RESULTS
The objective of this blinded, randomised study was to demonstrate the effica- After treatment, rectal temperature values, respiration and demeanour scores
cy of Resflor®, a combination of florfenicol and flunixin, against an experimental significantly decreased within 24 hours post-treatment with Resflor® and Draxx-
Mycoplasma bovis challenge in calves, in comparison to a negative control sa- in® plus flunixin, while they remained high for 7 days post-treatment with saline
line and a positive control tulathromycin associated with flunixin. (p<0.0001) (Fig. 1, 2 and 3). At both day 4 and day 7 post-treatment, clinical re-
covery with Resflor® was significantly superior to saline (p<0.0001) and non-infe-
MATERIALS AND METHODS rior to Draxxin® plus flunixin.
A total of 230 calves between 4 and 8 weeks of age, confirmed to be M. bovis The proportion of lungs with M. bovis lesions was 6.6% for Resflor®, 5.25% for
antibody and antigen negative, were challenged daily for 3 consecutive days by Draxxin® plus flunixin and 19.2% for saline (Fig. 4).
intratracheal deposition of 12 mL pure M. bovis culture (5.14x109 cfu, European
isolate 208/B01). Recovery of M. bovis from lung fluid was significantly lower for Resflor® than for
saline (2.27x105 cfu/mL versus 1.33x107 cfu/mL – p=0.0020) and for Draxxin®
For 4 days after challenge, calves meeting enrolment criteria (pyrexia >39.5°C plus flunixin (2.27x105 cfu/mL versus 2.53x106 cfu/mL – p=0.0120) (Fig. 5).
and abnormal respiration and/or depressed demeanour) were enrolled for treat-
ment with either saline (N=21), Draxxin® (tulathromycin) plus flunixin (N=64) or A significant difference in daily weight gain over the 7-day study was observed in
Resflor® (florfenicol-flunixin combination) (N=64). After treatment, animals were Resflor® and Draxxin® plus flunixin (mean 0.40 kg/day – p<0.0001), compared to
observed daily for seven days for clinical recovery (rectal temperature, respira- saline (0.13 kg per day – p=1.0000).
tion and demeanour). On day 7 post-treatment, animals were euthanized, lungs
collected for lesions scoring and lung lavage fluid collected for M. bovis bacteri- CONCLUSIONS
ology.
The combination of florfenicol and flunixin, Resflor®, provided an efficient clinical
and microbiological cure of M. bovis pneumonia in calves.
FIGURE 1. Effect of treatment on mean rectal temperature (°C). FIGURE 2. Effect of treatment on mean respiratory characters score.
FIGURE 3. Effect of treatment on mean demeanour score.
FIGURE 4. Percentage of lung with lesion at necropsy (day 7).
FIGURE 5. M. bovis recovery counts from lavage fluid at necropsy (day 7).
MSD Animal Health
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37 WBC 2016 (P04-004-116)
MSD ANIMAL HEALTH
Pulmonary lesions and clinical disease in Histophilus
somni-challenged calves treated with, either tildipirosin or
tulathromycin
Anthony W. Confer,a Jared D. Taylor,a Timothy A. Snider,a Mark Spireb
aDepartment of Veterinary Pathobiology, Oklahoma State University, Stillwater, OK 74078, USA; bMerck Animal Health, 35500 W. 91st St, De Soto, KS 66018, USA
INTRODUCTION TABLE 1 Day 1 Day 2 Day 3
Bovine respiratory disease (BRD) is the most severe cause of Experimental design
morbidity and economic losses in North American beef cattle.
Viruses, stress and bacteria such as Mannheimia haemolytica Day 0
and Histophilus somni are important in BRD.Recently, H.
somni are becoming increasingly resistant to tetracyclines, Clinical (twice): visual, respirations, Clinical (twice): Clinical (twice): BSL-2 barn - Clinical:
tulathromycin, enrofloxacin, and florfenicol. Tildipirosin temp, weight • visual, • visual, • visual,
(Zuprevo®, Merck Animal Health) is a long-acting macrolide Blood sample • respirations, • respirations, • respirations,
antimicrobial that recently became commercially available. Treatment groups • temperature • temperature • temperature,
The objective of this study was to compare in calves the 1. Tildipirosin 18% at 4 mg/kg • Sedation
efficacy of two macrolide antimicrobials, tildipirosin and (1 mL/100 lbs BW) Necropsy laboratory
tulathromycin (Draxxin®, Zoetis Animal Health) given at the 2. Tulathromycin at 2.5 mg/kg • Euthanasia
time of H. somni challenge and to examine their effects on (1.1 mL/100 lbs BW) – positive control • Necropsy
pulmonary lesions. 3. NaCl, 1 mL isotonic saline/100 lbs • Lung and other
BW – negative control
MATERIAL AND METHODS Challenge: Intrabronchial, 7 x 109 organ evaluations
CFU H. somni
Experimental design. Study design is shown in Table 1.
Cattle. Twenty-four, 2-3 month old, non-vaccinated Holstein TABLE 2
steers
Housing. Four calves per pen in a BSL-2 barn Body weights in kilograms (BW ± SD) at sorting and treatment/challenge, and necropsy. Weights per group were not significantly
Challenge. Intrabronchial with 7.0 x 109 CFU of virulent different among groups on Day 0 (p > 0.05). Days -3 and 0, body weights increased Following challenge, mean weight per group
H. somni strain 7735 (provided by Dr. Tom Inzana, Virginia dropped by 0.8 – 2.0 kg per group, but weight losses per group were not statistically different (p = 0.98).
Tech) which is susceptible to both antibiotics (Kansas State
University, Veterinary Diagnostic Laboratory). Group BW (kg) Day – 3, Sorting BW (kg) Day 0, treatment BW (kg) Day 3, necropsy
Clinical Examinations. Body temperature was measured at
0, 12, 24, 48, and 72 hours after treatment/challenge. Clinical Tildipirosin 84.3 ± 9.0 96.7 ± 9.8 95.2 ± 7.0 (- 1.5)*
scores included: General behavior, 0-4; Appetite at time of 98.5 ± 6.6 97.7 ± 5.9 (- 0.8)*
feeding, 0 – 3; Quality of respiration, 0 - 3. Tulathromycin 86.6 ± 4.5 94.1 ± 10.1 92.1 ± 6.8 (- 2.0)*
Post mortem examinations. Lung weight as a percentage of
body weight on the day of necropsy; Percent pneumonia. Saline 84.7 ± 6.6
Histopathologic evaluations. Lung Histopathology Score
(LHS), 0-4 for each lung; Necrosis score, 0-4 for each lung; Total * Change in weight (kg) between Days 0 and 3
LHS (maximum score of 12 per calf)
Total LHS = left lung LHS + right lung LHS + (necrosis scores TABLE 3
for both lungs)/2.
Bacterial culture scores. Left and right lungs were cultured to Rectal temperatures (mean ± SD) after challenge (˚C).Between 0 and 8 hours after challenge, rectal temperatures significantly
identify H. somni. Qualitative number of colonies cultured: 0 - 3 increased for each group (p < 0.05), but were not significantly different among treatment groups (p> 0.05).
for each lung.
Statistical analysis: Statistical comparisons by either non- Group Hours after challenge
parametric or general linear model (PROCMIXED, SAS 9.4)
including Kruskal-Wallis ANOVA, Mann-Whitney U with 0 8 24 48 72
a Bonferroni correction, and Kolmogorov-Smirnov test of
normality. Tildipirosin 39.4 ± 0.6 40.3 ± 0.7 39.1 ± 0.4 38.6 ± 0.9 39.2 ± 0.8
RESULTS Tulathromycin 39.1 ± 0.4 40.2 ± 0.2 38.8 ± 0.3 38.4 ± 0.4 38.9 ± 0.3
The development of body weights, rectal temperatures, clinical Saline 39.3 ± 0.2 40.3 ± 0.4 39.6 ± 0.8 39.2 ± 0.5 39.4 ± 0.5
and necropsy evaluations, lung pathology and histopathology
in the 3 treatment groups are shown in Table 2, 3, 4, 5, 6 and TABLE 4
Figure 1 and 2, respectively. Bacterial isolation scores were
significantly higher for saline-treated than for tildipirosin and Clinical evaluations. Clinical scores as means ± SD. Medians are in parentheses. Maximu m score 10 for times 0, 12, 24, & 48 hr.
tulathromycin -treated (p<0.05) calves. H. somni was not Clinical scores significantly increased for all groups by 8 hours after challenge were not significantly.different among groups (p > 0.05).
isolated from tildipirosin-treated calves.
Group Hours after challenge
CONCLUSIONS
0 8 24 48 72
Tildipirosin-treated calves had the lowest percent pneumonia.
Histopathological scores were similar for tildipirosin- and Tildipirosin 0.0 ± 0.0 5.1 ± 3.0* 3.3 ± 1.1* 1.4 ± 0.9 1.6 ± 1.5
tulathromycin-treated calves.
Necrosis was significantly less in macrolide-treated groups than Tulathromycin 0.0 ± 0.0 5.4 ± 2.5* 3.8 ± 1.6* 1.4 ± 0.9 1.0 ± 1.1
in control groups even when other pneumonic parameters were
similar. Saline 0.0 ± 0.0 3.3 ± 2.9* 2.8 ± 2.4* 1.8 ± 1.1 0.8 ± 1.0
Therefore, if necrosis can be avoided or reduced through
macrolide treatment, significant scarring of the lung can be * Clinical scores were significantly increased (p < 0.05) for all groups compared to Day 0 scores
avoided, and pulmonary function restored to normal or near
normal levels. TABLE 5
Necropsy evaluations Lung weights in grams (LW), percentage LW/BW at treatment, and percent pneumonia. Lung weight as a
percentage of body weight and percentage pneumonia were significantly lower for tildipirosin treated calves than for the saline-
treated calves (p < 0.05).
Group LW (g) % LW/BW at treatment % pneumonia
Tildipirosin 1000.8 ± 165.6 1.04 ± 0.2a 5.6 ± 6.9a
8.6 ± 8.9ab
Tulathromycin 1112.1 ± 180.7 1.18 ± 0.2ab 18.2 ± 9.9b
Saline 1278.1 ± 213.0 1.34 ± 0.2b
Differences in superscript letters indicate statically significant differences (p < 0.05).
TABLE 6
Lung histopathology scores (LHS ± SD); sum of lesion scores (0 = no lesion; 1 = Minimal; 2 = Mild; 3 = Moderate; 4 = Severe)
for right and left lung and necrosis score (idem). Lung necrosis was often severe in saline-treated calves. Right lung lesion,
necrosis, and LHS scores were significantly lower (p < 0.05) for tulathromycin- and tildipirosin-treated calves compared to saline-
treated calves (Table 5).
Group RLung** LLung** Necrosis*** Total LHS
Tildipirosin 2. 2 ± 0.5a 0.6 ± 0.7a 0.0 ±0.0a 2.8 ± 0.8a
Tulathromycin 2.4 ± 0.6a 0.5 ± 0.5a 0.0 ±0.0a 2.9 ± 1.0a
Saline 3.9 ± 0.2b 0.6 ±0.7a 2.9 ± 1.6b 7.4 ± 1.5b
Differences in superscript letters indicate statically significant differences (p < 0.05).
FIGURE 1A
Lung from a calf treated with tildipirosin
and challenged with H. somni. The lung
is essentially normal with mild edema in
the right caudal dorsal lobe.
1A FIGURE 1B
Lung from a calf treated with saline FIGURE 2 A BC
and challenged with H. somni. The right
caudal dorsal lobe is greatly swollen and
discolored. It was palpably firm and
on cut surface had extensive areas of
necrosis and inflammation. Note: The Histologic sections of lung from three calves (200X). Figure 2A. Tildipirosin-treated calf. Alveoli are collapsed and contain
intrabronchial method of challenge used primarily macrophages. There are small foci of neutrophils within several alveoli. Bronchioles are normal. Figure 2B. Saline-
treated calf. There is a fibrinosuppurative exudate within the bronchiole. Fibrin, numerous viable and necrotic neutrophils,
in this study resulted in lesions primarily and macrophages are present within most alveoli. Figure 2C. Saline-treated calf. Necrotic alveoli contain abundant protein-
1B in the right caudal dorsal lung lobes. rich edema fluid, fibrin, and leukocytes within their lumens. In other areas, alveolar structure is obscured by diffuse neutrophil
infiltrates and necrotic debris.
MSD Animal Health 38
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WBC 2016 (P04-004-108)
MSD ANIMAL HEALTH
Incidence and antibiotic susceptibility of non-viral pathogens
isolated from calves with BRD
Nijhoving G.H.1#, Kuijk H.1, Geurts C.P.H.2, Botvliet I.3
1MSD Animal Health, the Netherlands, 2D.A.P. Thewi, the Netherlands, 3B.V.L., the Netherlands,
#corresponding author: [email protected]
OBJECTIVES 100
Bovine Respiratory Disease (BRD) is a frequently 90
occurring disease in both dairy and veal calves.
This multifactorial disease complex is caused by 80
interactions between a wide range of pathogens,
the immune status of the calf and environmental % of farms pathogen isolated 70
influences. The pathogens involved in the complex 60
are viral or non-viral. The most common non-viral
BRD pathogens are Mannheimia haemolytica (M. 50
haemolytica), Pasteurella multocida (P. multocida),
Mycoplasma bovis (M. bovis) and Histophilus somni 40
(H. somni).
30
BRD has a major impact on performance and wellbeing 20
of the calf. Prevention of BRD is more important than
treatment. However, if BRD occurs, treatment should 10
be effective to control damage caused by this disease.
To ensure the efficacy of BRD treatment during 0 Pasteurella Mycoplasma Histophilus
outbreaks, both the presence and antibiotic sensitivity Mannheimia multocida bovis somni
of pathogenic bacteria should be determined. The haemolytica
objective of the study was to isolate BRD causing
non-viral pathogens from veal and dairy calves during FIGURE 1
disease outbreaks and to determine their antibiotic
sensitivity. Percentage of farms (n=178) where during BRD outbreaks Mannheimia haemolytica (n= 104; # samples: 297), Pasteurella multocida
(n= 159; # samples 572), Mycoplasma bovis (n= 125; # samples 403) and Histophilus somni (n=2; # samples 2) was isolated.
MATERIAL AND METHODS
Mannheimia haemolytica Pasteurella multocida
At the start of BRD outbreaks on 178 Dutch veal and 100
dairy farms between March 2013 and October 2015, % of bacterial strains sensitive
deep nasopharyngeal swabs were taken from calves, to antibiotic 75
two third of the samples were taken of veal calves and 50
a third of dairy calves. Calves with BRD symptoms 25
and not treated with antibiotics were enrolled in the
study. The inclusion criteria for BRD symptoms were 0
temperature, coughing, nasal and eye discharge and
position of the ears. Each symptom was scored on a Doxy/tetra
severity scale from 0 to 3 and calves should score at Florfenicol
least 5 points for inclusion.
TMPS
1103 calves were enrolled and sampled for AErmytoxhry/o/atymlpoi//tiplemn
bacteriological culture and antibiotic susceptibility
testing. The samples of each outbreak were inoculated Flumequine
directly on blood agar and chocolate agar plates Enro/marbo
and for mycoplasma inoculation on preheated PAM
and PAM+ plates. The plates were incubated under FIGURE 2
aerobic and micro aerophilic conditions. The inhibition
zones of these BRD bacteria were determined using Antibiotic susceptibility (%) of Mannheimia haemolytica isolates (n=297) and Pasteurella multocida isolates (n=572) during BRD
antibiotic sensitivity discs (paper discs) containing outbreaks (n=178 farms) for 7 different antibioics or antibiotic groups, stratified on antibiotic classification as first (green), second
doxycyclin/tetracyclin, erythromycin/tylosin/tilmicosin, (yellow) and third (red) choice.
florfenicol, trimethoprim sulfa, amoxicillin/ampicillin/
penicillin, flumequin and enrofloxacin/marbofloxacin CONCLUSIONS
and read according to the EUCAST tables. For M.
bovis no susceptibility testing was done, because the In conclusion, the decreased antibiotic susceptibility found in this study towards first choice
used method is not suitable for susceptibility testing antibiotics, suggests more emphasis should be put on the prevention of BRD. However, if BRD
of M. bovis. The choice of these antibiotic sensitivity occurs, florfenicol should be considered as a first choice treatment.
discs is based on the Dutch formulary for prudent use
of antibiotics, where the classification of the groups
(first, second and third choice) is based on antibacterial
spectrum and the effects on selection pressure
causing antibiotic resistance.
RESULTS
The incidence of isolated BRD pathogens on farm
are shown in Figure 1. Antibiotic susceptibility for
both M. haemolytica and P. multocida showed major
differences; low sensitivity for first choice antibiotics
(doxycyclin/tetracyclin, erythromycin/tylosin/tilmicosin,
and trimethoprim sulfa), but high susceptibility towards
florfenicol (Figure 2).
MSD Animal Health
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39 WBC 2016 (P04-004-123)
MSD ANIMAL HEALTH
Fipronil and Fluazuron: efficacy of a novel parasiticide combination
against Dermatobia hominis and Cochliomyia hominivorax in
naturally infested cattle
Lopes, W.D.Z.1; Chiummo, R.M.2; Botelho, E.M.2; Vettorato, L.F.2
1Tropical Pathology Institute and Public Health, University of Goiás, Brazil; 2MSD Animal Health, São Paulo, Brazil
INTRODUCTION
Furuncular myiasis or dermatobiosis, caused by infestation of histotropic
larvae of Dermatobia hominis grubs, often occurs in the subcutaneous tissue
of warm blood animals, from Mexico to temperate regions of Argentina.
The movements of D. hominis larvae cause pain, restlessness and irritation,
resulting in meat and milk production losses and depreciation of leather.
Cochliomyia hominivorax screwworm (Coquerel, 1858) causes primary
cutaneous myiasis in domestic and wild animals, resulting in reduction in
productivity, weight loss, growth delay, expenditure with chemicals and even
death of animals by septicemia.
The efficacy of a new pour-on formulation comprising the anti-parasiticides
Fipronil (1.25 mg/kg) + Fluazuron (2.5 mg/kg) was evaluated in two field
studies against Dermatobia hominis (Experiment 1) and Cochliomyia
hominivorax (Experiment 2) in naturally infested cattle.
MATERIAL AND METHODS FIGURE 1
• Experiment 1 was carried out from the Southeast region of São Paulo State, Results of Experiment 1. Efficacy (percentage of larvae killed after treatment compared to the untreated
Brazil. Twenty animals were selected from 40 Canchim female cattle, not control group) after treatment on day 0 with the parasiticide combination fibronil (1.25 mg/kg) and fluazuron
treated with endectocide for 120 days, naturally infested with D. hominis (2.5 mg/kg) of Dermatobis hominis in naturally infested cattle (treatment n=10, control n=10).
larvae. Bovines were randomized into two experimental groups of ten
animals each, using a randomized complete block design based on the From the 1st to 10th DPT, the Fipronil + Fluazuron combination reached 100% of
mean counts of D. hominis larvae nodules on Day -1, present on the entire preventive effectiveness, preventing G1 animals to develop active myiasis in all
body surface of the animals (Table 1). lesions. The animals in the control group showed active myiasis from the 2nd to
the 5th DPT, after which they had to be treated for animal welfare reasons (using a
• Experiment 2 was carried out on a farm located at the municipality of commercially available topical organophosphate)(Table 2).
Abadia dos Dourados, Minas Gerais State-Brazil. Twelve crossbred calves
were divided into two groups of six animals each. G1 was treated with * Tick Gard – MSD Animal Health.
the Fipronil + Fluazuron combination and GII was kept as a control. On NR = Note realized: the bovines had to be treated for animal welfare reasons
day 0 (before treatment), each animal was subjected to two circular skin
incisions (one on each side of the body), measuring about 4cm in diameter TABLE 2
each, in the region between the infra-spinal cavity and the dorsal scapula
border. This procedure was performed after the animals receive sedatives Results from Experiment 2. Therapeutic efficacy of animals (n=6) treated with fibronil (1.25 mg/kg) and
(dihidrotiazine hydrochloride) and local anesthesia (lidocaine 2%). Right after fluazuron (2.5 mg/kg) compared to the non-treated control (n=6), against Cochliomyia hominivorax larvae
the aforementioned surgical procedure, the treatment of animals from G1 causing myiasis in naturally infested cattle.
was carried out. Immediately after treatments, the two groups were kept in
separate adjacent pastures, allowing the natural exposure of lesions to C.
hominivorax infestations.
RESULTS
Against Dermatobia hominis grubs, the combination of Fipronil and Fluazuron
demonstrated efficacy higher than 90% from the 7th to the 35th day post-
treatment (DPT), showing 99.5% of efficacy on 21 and 28 DPT. In all post-
treatment dates, the D. hominis larvae counts were statistically lower (p ≤
0.05) in the animals from G1 when compared to the control animals GII.
CONCLUSIONS
With the results of these two studies it is possible to infer that the new
parasiticide formulation containing both Fipronil (1.25 mg/kg) + Fluazuron
(2.5mg / kg), in the dose and route applied, proved high therapeutic efficacy
against the tick larvae of Dermatobia hominis larvae and preventive efficacy
against Cochliomyia hominivorax in naturally infested cattle.
Experiment Groups No. of Cattle Parasiticide Treatment Route of Administration Dosage
mL/kg bw mg/kg
I 10 Control (saline) Pour-on 1/10 -
1 10 Fipronil + Fluazuron* Pour-on
Pour-on 1/10 1.25 + 2.5
II Pour-on
I 6 Control (saline) 1/10 -
6 Fipronil + Fluazuron*
2 1/10 1.25 + 2.5
II
* Tick Gard – MSD Animal Health
TABLE 1
Experimental Groups and Treatment Design used in both experiments.
MSD Animal Health 40
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P04-004-073)
MSD ANIMAL HEALTH
Pharmacokinetic evaluation of a new combination of Fipronil and
Fluazuron topically administered (pour-on) in cattle
Lopes, W.D.Z.1; Chiummo, R. M.2; Botelho, E.M.2; Vettorato, L.F.2
1Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, GO, 74605050, Brazil;
2MSD Animal Health, São Paulo-SP, Brazil.
INTRODUCTION TABLE 1
The activity of ectoparasiticides compounds can be affected by the Plasma mean concentration of fipronil administered at dose of 1.25mg/kg in cattle.
concentration and exposure time of these assets with the parasites.
TABLE 2
Thus, changes in the formulations can cause significant changes in the
efficacy of the drug, thus demonstrating the need to carry out studies of Plasma mean concentration of fluazuron administered at dose of 2.5mg/kg in cattle.
new pharmacological formulations developed.
Because of this, this study aimed to determine the pharmacokinetic
parameters of a new combination of Fipronil and Fluazuron, administered
to cattle via pour-on at a single dose of 1ml / 10kg body weight
(corresponding to 1.25 mg/kg of Fipronil and 2.5mg/kg of Fluazuron).
MATERIAL AND METHODS
Eight crossbred animals with 14 to 24 months old (four males and
four females) were selected. Animals were not exposed to any other
parasiticide at least 60 days prior to the start of the study. All animals
received the combination product on day 0 of the study. A 10 mL
blood sample was collected by jugular vein from all animals at time 0
(immediately before treatment) and with 3, 6, 12, 24, 48, 60 hours and
3, 4, 6, 8, 10, 12, 14, 17, 21, 28, 35 and 42 after treatment. Up to two
hours after each collection, samples were centrifuged for 15 minutes at
1.200G (gravity). The separated plasma was placed into two siliconized
Eppendorf® tubes (1.5 mL each) pre-identified and stored at -20°C until
transportation to the laboratory. The analysis of Fipronil and Fluazuron
were performed by liquid chromatography attached to a mass detector
(LC-MS / MS).
RESULTS
The Fipronil maximum concentration was 73.7 g / L (Cmax) with 2.5 days
after administration of the product (Tmax). The AUC42d for Fipronil was
35699.4μg-hr / L) and the half-life (T1/2) was between 17 and 21 days after
treatment (C1/2 = 36.85μg / L). For Fluazuron, the maximum concentration
in the plasma was 26.1μg / L (Cmax) occurred after six days after the
product was administered (Tmax). The area under the curve (AUC42d) was
10472.6 4μg-hr / L. The half-life (T1/2) was 13.05μg / L (C1 / 2) obtained
between 21 and 28 days after administration.
CONCLUSIONS
Given the results, it can be inferred that Fipronil showed peak
concentrations (2.5 days) earlier than Fluazuron (6 days), followed by a
gradual decrease of both active ingredients until the last observation day
(42 days post-treatment).
FIGURE 1
*Tick Gard – MSD Animal Health, São Paulo, Brazil.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
41 WBC 2016 (P04-004-110)
MSD ANIMAL HEALTH
Therapeutic and residual efficacy of a novel Fluazuron combination against
Rhipicephalus (Boophilus) microplus ticks in naturally infested cattle
Lopes, W.D.Z.1; Chiummo, R. M.2; Botelho, E.M.2; Vettorato, L.F2
1Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, GO, Brazil; 2MSD Animal Health, São Paulo-SP, Brazil
INTRODUCTION
The southern cattle tick,
Rhipicephalus (Boophilus)
microplus, is the most
important ectoparasite
of cattle in Brazil. This
tick is a vector for various
diseases and, moreover,
Source: Welber Daniel Zanetti Lopes poses as a major obstacle
to animal productivity
in tropical and sub-tropical areas of the world where the
species is endemic.
Controlling this ectoparasite is a key factor in improving
production, but the efforts to combat it on most cattle
farms are performed incorrectly, characterized by excessive
and disordered use of therapeutic approaches.
In this context, two experiments were conducted to
evaluate the efficacy of a new formulation containing
Fipronil (1.25mg/kg) + Fluazuron (2.5mg/kg), administered
once, via pour-on, against Rhipicephalus (Boophilus)
microplus in naturally infested cattle.
MATERIAL AND METHODS FIGURE 1
Two experiments were carried out to assess the acaricidal Experiment I, Aquidauana-MS, Brazil. Efficacy of fipronil 1.25mg/kg + fluazuron 2.5mg/kg against R. (B.) microplus in crossbreed steers
efficacy of fipronil 1.25mg/kg + fluazuron 2.5mg/kg: naturally infested.
Experiment I – Natural infestation: Crossbreed steers.
Conducted in Aquidauana-MS, Brazil.
Experiment II – Natural infestation: Simmental cattle. São
José do Rio Pardo-SP, Brazil.
In each experiment, 20 animals were randomly divided
into 2 groups of 10, based on the average counts of female
ticks (between 4.5 to 8.0mm) from each animal on three
consecutive days prior to treatment (days -3, -2 and -1), as
described in the Table 1.
The therapeutic and residual efficacy of Fibronil + Fluazuron
were assessed on days 1, 3, 7, 14, 21, 28 and weekly until
56 days post-treatment (DPT). Efficacy was expressed in
percentage and defined as:
Efficacy percentage = 1- Ta x Cb x 100
Tb x Ca
In this equation, Ta represents the mean number of 4.5-
8 mm female ticks counted on treated animals after FIGURE 2
medication; Tb is the mean number of 4.5 - 8 mm female
ticks counted on treated animals during the three days Experiment II, São José do Rio Pardo-SP, Brazil. Efficacy of fipronil 1.25mg/kg + fluazuron 2.5mg/kg against R. (B.) microplus in Simmental
that preceded treatment; Ca is the mean number of 4.5 cattle naturally infested.
- 8 mm female ticks counted on the control group after
the experiment was initiated; and Cb is the mean number
of 4.5 - 8 mm female ticks counted on untreated animals
(control) during the three days that preceded treatment.
In both experiments, the susceptibility of the field strain to
fluazuron and fipronil was unknown.
RESULTS TABLE 1
In experiment I, the combination of Fipronil + Fluazuron Experimental design used in both experiments.
obtained efficacy higher than 95% from the 3rd to the
35th DPT (96.26%, 100.00%, 99.58% and 97.81% Experiment Group No. of cattle Treatment (pour-on) Dose (mg/kg bw)
respectively on days 3, 7, 14, 21, 28 and 35 post- Control (saline solution) -
treatment). In this experiment, the efficacy was already I and II I 10
higher than 90% (93,32%) at 2 DPT. The efficacy of II 10 Fibronil + Fluazuron* 1.25 + 2.5
Fipronil + Fluazuron remained higher than 80% until 42
DPT (88.79%) (Figure 1). *Tick Gard – MSD Animal Health, São Paulo, Brazil.
In experiment II, the combination of Fipronil + Fluazuron CONCLUSIONS
obtained efficacy higher than 95% from the 7th to the
35th DPT (96.32%, 96.40%, 97.22%, 97.08% and 96.28% Based on the results obtained in these 2 experiments, the new parasiticide combination
respectively on 7, 14, 21, 28 and 35 DPT) (Figure 2). In of Fipronil 1.25mg/kg + Fluazuron 2.5mg/kg administered once via pour-on, showed high
this experiment, the efficacy was still higher than 90% efficacy against Rhipicephalus (Boophilus) microplus ticks in naturally infested cattle.
(93.32%) at 42 DPT.
In both studies, the R. (B.) microplus counts on the
Fipronil + Fluazuron treated animals were significantly
lower (p ≤ 0.05, t-test) than the control animals from the
3rd to the 56th DPT.
MSD Animal Health 42
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P04-004-111)
MSD ANIMAL HEALTH
Comparative therapeutic and residual efficacy of a novel parasiticide
combination of fipronil and fluazuron against Rhipicephalus
(Boophilus) microplus ticks in artificially infested cattle
Lopes, W.D.Z.1; Chiummo, R. M.2; Botelho, E.M.2; Vettorato, L.F.2
1Instituto de Patologia Tropical e Saúde Pública, Universidade Federal de Goiás, Goiânia, GO, Brazil; 2MSD Animal Health, São Paulo-SP, Brazil
INTRODUCTION
Source: National Institute of Livestock Technology Rhipicephalus
(www.inta.gov.ar) (Boophilus) microplus
tick is considered
the most important
cattle ectoparasite
in Brazil, due to the
direct harm it causes
to their health,
significantly restricting
animal productivity
and causing great
economic losses.
The control of this ectoparasite is undeniably necessary, and it
is done with synthetic acaricides.
In this context, the therapeutic efficacy and residual activity of
a new pour-on formulation containing Fipronil (1.25 mg /kg) +
Fluazuron (2.5mg/kg), * was assessed against R. (B.) microplus
in cattle submitted to artificial infestations.
MATERIAL AND METHODS FIGURE 1
The experiment was conducted in the city of Abadia dos Efficacy (%) of 3 different parasiticide pour-on formulations (see Table 1) against R. (b.) microplus ticks in artificially infested
Dourados, Minas Gerais State, Brazil. 24 crossbreed steers cattle.
were randomized in four groups of six animals each, based on
the average counts of engorged female ticks detached from TABLE 1
each animal on three consecutive days prior to treatment (days
-3, -2 and -1), as described in the Table 1. Experimental design of the study.
The therapeutic and residual efficacy against R. (B.) microplus Group No. of cattle Treatment (pour-on) Dose (mg/kg)
was evaluated by daily counts of all female ticks detached from Fipronil + Fluazuron* 1.25 + 2.5
each animal, from the 1st to 60th day post treatment (DPT). I6 1.0
Efficacy was expressed in percentage and defined as: 2.5
II 6 Fipronil** -
Efficacy percentage = 1- Ta x Cb x 100
Tb x Ca III 6 Fluazuron**
In this equation, Ta represents the mean number of partially IV 6 Control (saline solution)
engorged female ticks counted on treated animals after
medication; Tb is the mean number of partially engorged *Tick Gard – MSD Animal Health, São Paulo, Brazil.
female ticks counted on treated animals during the three days **Commercial product purchased in the Brazilian market
that preceded treatment; Ca is the mean number of partially
engorged female ticks counted on the control group after CONCLUSION
the experiment was initiated; and Cb is the mean number
of partially engorged females counted on untreated animals Based on the results from this experiment, the new parasiticide combination of Fipronil
(control) during the three days that preceded treatment. 1.25mg/kg + Fluazuron 2.5mg/kg administered once via pour-on, showed higher mean
therapeutic efficacy and longer residual protection against Rhipicephalus (Boophilus)
The susceptibility of the field strain to fluazuron and fipronil was microplus ticks in artificially infested cattle when compared to standalone formulations of
unknown. Fipronil and Fluazuron.
RESULTS
In this study, treatment group I showed mean therapeutic
efficacy (1 to 23 DPT) of 95.8% while treatment group II and
III presented respectively 94% and 89%. Treatment group I
presented efficacy ≥90% from the 4th to 45th DPT, with efficacy
≥95% in 30 counting days during this period. On the other
hand, treatment group I and III presented, respectively, efficacy
≥90% from the 4th to 40th DPT and from the 5th to the 39th DPT
(31 and 27 counting days with efficacy higher than 95%). The
efficacy of such formulations were above 80% until 57 (group
I), 42 (group II) and 48 (group III) DPT (Figure 1).
The statistical comparison (t-test) showed that group 1 mean
counts were significantly lower (p≤0.05) than the control group
counts from the 3rd DPT until the end of the study (60 DPT). The
animals from group 2 presented statistically lower mean counts
compared to the control group (p≤0.05) from 2nd to 46th DPT
while this difference was significant for group 3 treated animals
from 4th to 56th DPT.
MSD Animal Health
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
43 WBC 2016 (P04-004-112)
MSD ANIMAL HEALTH
Antimicrobial susceptibility of staphylococcus aureus isolated
in bovine mastitis in France
Sylvain BAREILLE1, Frédéric LEBOEUF1, Guillaume LEQUEUX2, Jantijn SWINKELS3
1MSD Santé Animale, 7 rue Olivier de Serres, Beaucouzé (49), France, 2ISAE 35, BioAgroPolis – 10 rue Claude Bourgelat, Javené (35), France, 3MSD Animal Health, 556 Morris Avenue, Summit, NJ 07901, USA
OBJECTIVES All S. aureus isolates were sensitive to cloxacillin, nafcillin, cefazolin,
cephapirin, cephalonium and amoxicillin-clavulanic acid. Antibiotic resistant
To determine the minimal inhibitory concentrations (MIC) of antimicrobials strains were observed for penicillin G (n=17, 21%), cephalexin (n=2, 3%)
for Staphylococcus aureus (S. aureus) from the milk of dairy cattle in and cefquinome (n=2, 3%). This percentage (21%) of S. aureus strains
France. Getting new data for this topic is relevant because S. aureus is resistant towards penicillin G is lower than the 30% mentioned in the
one of the most frequently isolated bacteria in milk samples from sub- RESAPAPH report.
clinical cases of bovine mastitis in France (about 18% excluding sterile or
contaminated samples). Finally, these data may help reaching a better and The antimicrobial susceptibility of S. aureus towards the different
responsible use of antibiotics and possibly contribute to the reduction of cephalosporins varies significantly. For example, the MIC90 of cephalonium
antibiotic use in France of 25% from 2012 to 2017. is only 0.125μg/mL towards S. aureus, when MIC90 of cephalexin is 4μg/
mL.
MATERIAL AND METHODS
There is a high and homogeneous level of antimicrobial susceptibility of
The susceptibility of 80 isolates of S. aureus to antibiotics was assessed S. aureus towards gentamicin (all MIC ≤ 0.25μg/mL). But, the antimicrobi-
by the standard agar dilution method. These strains were isolated using al susceptibility of S. aureus towards tylosin is just below the breakpoint
milk samples from cows affected by clinical or sub-clinical mastitis. (MIC90 = 2 μg/mL). A bimodal repartition of the MIC of S. aureus strains
They were chosen at random from all milk samples received in 2013 towards tylosin was seen, as previously described in a German study, with
by ISAE 35 laboratory from all over France and the strains tested were 2 strains (3%) showing a MIC of tylosin >256 μg/mL. S. aureus resistance
epidemiologically unrelated (eg not coming from the same episode of towards penicillins G is mainly due to the presence of the blaZ gene, cod-
disease or the same herd - guideline EMEA/CVMP/627/01). ing for a β-lactamase. The blaZ gene is frequently (30%, RESAPAPH report)
detected in the strains of S. aureus isolated in France. Because the differ-
A MIC-determination was performed using the agar dilution method ence between phenotypic resistance (MIC determination) and genetic re-
according to the standard method (standard CLSI VET01-A4). Twelve sistance is mainly due to the inducible expression of this gene, it seems
antimicrobial agents – penicillin G, cloxaxillin, nafcillin, cephalexin, that resistance is only observed if bacteria have previously been in contact
cefazolin, cephapirin, cephalonium, cefquinome, amoxicillin-clavulanic acid, with a β-lactam.
tylosin, gentamicin, and enrofloxacin – were investigated because they are
frequently used in the field.
RESULTS
The following table shows the current MIC of antimicrobials for
Staphylococcus aureus from the milk of dairy cattle in France.
TABLE
Minimum inhibitory concentration (MIC, μg/ml) of 80 isolates of S. aureus from milk samples from cows affected by clinical or sub-clinical mastitis to 12 different
antimicrobials agents.
MIC in µg/mL ≤0.06 0.06 ≤0.125 0.125 ≤0.25 0.25 0.5 1 2 4 8 >8 16 32 64 128 256 >256 MIC50 MIC90
cephalonium 70 10 ≤0.06 0.125
gentamicin 80 ≤0.25 ≤0.25
cephapirin 45 33 2 ≤0.125 0.25
enrofloxacin 25 30 22 1 1 1 0.125 0,25
cloxaxillin 10 62 7 1 0.25 0.25
penicillin G 56 7 87 2 ≤0.06 0.5
amoxicillin- 42 17 10 3 1 1 0.125 0.5
clavulanic acid
nafcillin 3 49 23 3 2 0,25 0,5
cefazolin 2 61 16 1 0.25 0.5
cefquinome 5 17 28 28 2 0.25 0.5
tylosin 2 29 35 12 21 2
cephalexin 1 4 42 29 2 2 24
Peer reviewed breakpoint for S. aureus. A breakpoint is the minimum inhibitory concentration of an antibiotic above which S. aureus is considered resistant and
below which S. aureus is considered susceptible to the antibiotic.
CONCLUSIONS
This in vitro study shows that the resistance of S. aureus towards penicillin G is widely observed and the antimicrobial susceptibility of S. aureus towards the
macrolides is not optimal (high MIC, and some strains are resistant towards this antibiotic group). Although in vitro MIC studies do not always predict in vivo
treatment success, based on these MIC data alone, the preferred antibiotics for treatment of S. aureus are cephalonium, gentamycin, cephapirin, cloxacillin,
nafcillin, cefazolin, amoxicillin-clavulanic acid.
MSD Animal Health 44
Copyright ©2016 Intervet International B.V, also known as MSD Animal Health. All rights reserved.
WBC 2016 (P04-004-113)
MSD ANIMAL HEALTH
Effect of two different bovine somatotropins on milk production
and reproduction of dairy cows in Brazil
Henderson Ayres1, Marcilio Nichi2, Sandro L. Viechnieski3, Pietro Sampaio Baruselli2, Rodrigo Almeida4
1MSD Animal Health, São Paulo, Brazil; 2Department of Animal Reproduction of USP, São Paulo, SP, Brazil; 3Star-Milk Farm, Céu Azul, PR, Brazil; 4Universidade Federal do Paraná, PR, Brazil
1,20 Group 1 Treat: P < 0.01
OBJECTIVE MATERIAL ANDGroup 2 METHODS Day: P < 0.01
The objective of this trial was to evaluate milk yield and assess the reproductive Treat x day: P < 0.01
outcomes of a high-producing dairy cattle herd in response to two commercial
sources of bovine somatotropin (bST) administered subcutaneously at a dose of Eight1y,1H5olstein cows from a commercial farm located in the south region
500 mg, 16 times with a 14 days interval. Relative milk yeild, kg/day
of Brazil, averaging 1.76 ± 1.16 lactations, 77.1 ± 21.9 days in milk (DIM) and
37.5 ± 6.5 kg/d of milk were randomly assigned to one of 2 treatment groups.
Grou1p,110 was treated with 500 mg bST (Boostin, MSD Animal Health) the
other group, (group 2) was treated with 500 mg of bST (Lactotropin, Elanco)
1,05
MILK PRODUCTION 1,20 1,00 Group 1 (C)
1,20 Group 2
Group 1 Treat: P < 0.01
1,15 0,95 Group 2
50 Group 1 Treat: P = 0.79 0 Day: P < 0.01
50 Group 2 Day: P < 0.01 2 TTrreeaatt:x day: PP << 00..0011
45 Group 1 TTrreeaatt:x day: PP <= 00..0719 1,15 Day: P < 0.01
45 Group 2 Day: P < 0.01
40 Treat x day: P < 0.01 RelatRiveleatimvilek ymileilkdy,eiklgd/,dkagy/day1,10 Treat x day: P < 0.01 16
40 X 1.000 SCC, per mL of milk 1,10 4 6 8 10 12 14
35
AverAavgeeraMgilek yMileikldy,eiklgd/,dkagy/day 35 Day after administration
30
30 UDDER HEALTH
25
25 1,05 800 Treat: P = 0.31
20 1,05 600
20 0 Group 1 Day: P = 0.13
1,00 Group 2
0 1,00 Treat x day: P = 0.58
(A) 0,95 2 4 6 8 10 12 14 16
(A) 0,95 0 400 4 Day6 after ad8ministra1t0ion 12 14 16
50 100 150 200 250 0 2 Day after administration
200 250
50 Days afte1r0a0d 1st Adm1in5i0stration
Days after ad 1st Administration 200
42 Group 1 Treat: P < 0.01 (D)
42 Group 2 Day: P < 0.01
Group 1 TTrreeaatt:x day: PP << 00..0011 0
40 Group 2 Day: P < 0.01 -2 0 2 4 6 8 10
40 Treat x day: P < 0.01 Week after 1st administration
AverAavgeeraMgilek yMileilkdy,eiklgd/,dkagy/day 38 FIGURE 2
38
CLINICAL MASTITIS: Of the 80 animals in 224 trial days, 17 animals had clinical mastitis, of which 8
36 cases in treatment group 1 and 9 cases in treatment group 2.
36
(B) TABLE 1
34 (B)
34 Average reproductive outcomes of 80 cows in a high producing herd in response to treatment with 2
2 4 6 8 10 12 14 16 different commercial sources of bovine somatotropin (bST).
0 2 4 Day6 after ad8ministra1t0ion 12 14 16
0
Day after administration
Treatment Protocol Group 1 Group 2 P
1,20 Group 1 Treat: P < 0.01 DIM 74.9 79.4 > 0.20
1,15 Group 2 Parity 1.75 1.78 > 0.20
Day: P < 0.01 PR at the initial 2.5% 12.5% > 0.20
Treat x day: P < 0.01
Relative milk yeild, kg/day 1,10 PR at the end 50.0% 55.0% 0.32
1,05 # of Services 3.45 ± 0.54 3.49 ± 0.40 0.93
ICP 129.7 ± 22.9 130.4 ± 17.1 0.97
1,00 (C)
DIM – Days in Milk; PR – pregnancy rate; # – numbers; ICP – interval (days) between calving and
pregnancy
0,95
0 2 4 6 8 10 12 14 16
Day after administration
FIGURE 1
Average daily milk yield (A), average weekly milk yield (B), average relative milk yield (C) and average somatic cell
ecoacuhntrsep(Dre) soefnatihnig8g0htr0-eparotmduecnint gwditahiGrayrodcuiafpftetl1reenhtercdom(nm=8e0rc) iiDTanlraersyeao:stu:procnesoef tboPoP2v==intr00ee..a13st31ommeanttogtrroopuipns(b(GSTro).up 1 and Group 2)
Group 2 Treat x day: P = 0.58
600 X 1.000 SCC, per mL of milk
CONCLUSION
This study show40s0that in treatment group 1, milk production was significantly
higher than in treatment group 2. No difference in udder health, nor in
reproduction outcomes was found between treatment group 1 and 2.
200
(D)
MSD Animal Hea0lth-2 0 2 4 6 8 10
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Week after 1st administration
45 WBC 2016 (P04-004-115)
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