Beruflich Dokumente
Kultur Dokumente
a r t i c l e i n f o a b s t r a c t
Article history: Four experiments evaluated the effects of vaccination against bovine herpesvirus-1 (BoHV-
Received 20 November 2012 1), bovine viral diarrhea virus (BVDV), and Leptospira spp. on reproductive performance of
Received in revised form
lactating dairy cows without (experiments 1, 2, and 3) or with previous vaccination against
24 December 2012
these diseases (experiment 4). Cows were assigned to a fixed-time AI protocol (FTAI; d
Accepted 29 December 2012
Available online 10 January 2013 −11 to 0) in all experiments, as well as AI 12 h upon estrus detection in experiment 3.
Pregnancy status was determined with transrectal ultrasonography on d 30 and 71 (d 60
Keywords: for experiment 3) after AI. Pregnancy loss was considered in cows pregnant on d 30 but
Dairy cows non-pregnant on the subsequent evaluation. In experiment 1, 853 cows received (VAC)
Reproductive diseases or not (CON) vaccination against BoHV-1, BVDV, and Leptospira spp. at the beginning of
Reproductive performance the FTAI (d −11) and 30 d after AI. Pregnancy loss was reduced (P = 0.03) in VAC cows
Vaccine compared with CON. In experiment 2, 287 cows received VAC or CON 30 d prior to (d −41)
and at the beginning (d −11) of the FTAI. Pregnancy rates on d 30 and 71 were greater
(P ≤ 0.03) in VAC cows compared with CON. In experiment 3, 1680 cows with more than
28 d in milk were randomly assigned to receive VAC or CON with doses administered 14 d
apart, and inseminated within 15–135 d after the second dose. Pregnancy rates on d 30 and
60 were greater (P ≤ 0.02) in VAC cows compared with CON. In experiment 4, 820 cows
received (REVAC) or not (CON) revaccination against BoHV-1, BVDV, and Leptospira spp. at
the beginning of the FTAI protocol (d −11). Pregnancy rates and loss were similar (P ≥ 0.54)
between treatments. Hence, vaccinating naïve cows against BoHV-1, BVDV, and Leptospira
spp. improved reproductive efficiency in dairy production systems, particularly when both
doses were administered prior to AI.
© 2013 Elsevier B.V. All rights reserved.
0378-4320/$ – see front matter © 2013 Elsevier B.V. All rights reserved.
http://dx.doi.org/10.1016/j.anireprosci.2012.12.011
M.H.C. Pereira et al. / Animal Reproduction Science 137 (2013) 156–162 157
development (Grooms, 2004; Grooms et al., 2007), whereas PGF2␣ treatment (12.5 mg im of Dinoprost; Pfizer Animal
the bovine herpesvirus-1 (BoHV-1) that causes IBR is Health) on d −4, estradiol cypionate treatment (1.0 mg im
known to directly impair ovarian function and embryo of ECP; Pfizer Animal Health) in addition to CIDR removal
quality (Miller and Van der Maaten, 1986; Kelling, 2007). on d −2, followed by fixed-time AI on d 0. None of these
These pathogens and respective diseases are present in operations had a history of vaccinating the cowherd against
cattle herds worldwide (Rebhun, 1995), thus impacting IBR, leptospirosis, and BVD. Within each operation, cows
reproductive and overall efficiency of the global dairy were randomly assigned to receive (VAC; n = 426) or not
industry. As an example, seroprevalence for BoHV-1, Lep- (CON = 427) vaccination against IBR, BVD, and leptospiro-
tospira spp., and the BVDV, as well as the incidence of IBR, sis (5 mL im of CattleMaster 4+L5, Pfizer Animal Health,) at
leptospirosis, and BVD are increased in Brazil (Takiuchi the beginning of the FTAI protocol (d −11) and 30 d after
et al., 2001; Flores et al., 2005; Junqueira et al., 2006). AI. Hence, the objective of this experiment was to evalu-
Management techniques to prevent pregnancy loss in ate the effects of vaccination against IBR, leptospirosis, and
dairy herds, such as hormonal manipulation, thermal com- BVD, with both doses administered when cows were han-
fort, and nutritional management are increasingly being dled for reproductive management, on pregnancy rates and
implemented into dairy systems worldwide (Lucy, 2001). pregnancy losses in dairy operations that did not have a
Conversely, immunization strategies developed to reduce history of vaccinating the cowherd.
the impact of reproductive diseases, such as vaccination On d −11, cows were (average ± SEM) 111 ± 3 d in milk
against IBR, leptospirosis, and BVD, do not receive proper (DIM), producing 21.3 ± 0.2 kg of milk, and body condition
attention (Littel-Van der Hurk, 2006). Recent research from score (BCS) of 2.89 ± 0.01 (Wildman et al., 1982). Pregnancy
our group demonstrated that vaccination against repro- status was verified by detecting a fetus via transrectal
ductive diseases increased overall reproductive efficiency ultrasonography (Aloka SSD – 500 with a 7.5 MHz linear-
in Brazilian beef herds (Aono et al., 2012), whereas few array transrectal transducer, Tokyo, Japan) on d 30 and 71
research studies directly evaluated the effects of such vacci- after AI. Any cow diagnosed as pregnant on d 30 and then
nation programs on reproductive efficiency in dairy cattle. non-pregnant on d 71 was designated as having under-
Hence, the objective of the present study was to evaluate gone pregnancy loss. Blood samples were collected from
the adoption of vaccination programs against IBR, BVD and a subsample of CON cows from 17 operations (n = 84) on
leptospirosis on pregnancy rates and pregnancy losses in d −11 for determination of serological profile for BoHV-1,
commercial dairy operations. BVDV, and Leptospira spp. infections. Blood samples were
randomly collected from an average of 5 females per oper-
2. Materials and methods ation, being 4 multiparous and 1 primiparous cow. Samples
were collected via coccygeal vein or artery into com-
All experiments described herein were conducted in mercial blood collection tubes (Vacutainer, 10 mL; Becton
commercial dairy operations located in Minas Gerais and Dickinson, Franklin Lakes, NJ), placed on ice immediately,
Paraná, Brazil. All animals utilized were cared for in accor- maintained at 4 ◦ C for 24 h, and centrifuged at 3000 × g for
dance with the practices outlined in the Guide for the Care 10 min at room temperature for serum collection. Serum
and Use of Agricultural Animals in Agricultural Research was stored at −20 ◦ C until further analysis. Neutralizing
and Teaching (FASS, 1999). Within each operation, cows antibodies against BoHV-1 and BVDV were detected by
were managed similarly independent of vaccination treat- virus neutralization test in Madin–Darby bovine kidney
ments, following existing nutritional, reproductive, and cells (Ferreira et al., 2005) and 100 tissue culture infec-
health-related procedures for each operation. tious dose50 of Los Angeles and NADL strains of BoHV-1 and
BVDV, respectively (Pilz et al., 2005). Detection of aggluti-
2.1. Vaccine nant antibodies against leptospirosis was conducted using
microscopic agglutination test (Ryu, 1970). The criteria for
The vaccine utilized in all experiments (CattleMaster seropositive animals were titers ≥ 8 for BoHV-1, ≥16 for
4+L5, Pfizer Animal Health, São Paulo, Brazil) was a freeze- BVDV, and ≥100 for Leptospira spp. (Ryu, 1970; Ferreira
dried preparation containing a chemically altered live et al., 2005; Pilz et al., 2005).
strain of BoHV-1, inactivated cytopathic and noncytopathic
BVDV strains, and cultures of five Leptospira spp. serovars 2.3. Experiment 2
(L. canicola, L. grippotyphosa, L. hardjo, L. icterohaemor-
rhagiae, and L. pomona), with the inclusion of aluminum A total of 287 lactating Gir × Holstein cows (multi-
hydroxide as adjuvant. parous, n = 212; primiparous, n = 75) originated from 28
operations were assigned to the FTAI protocol described
2.2. Experiment 1 in experiment 1. None of these operations had a history
of vaccinating the cowherd against IBR, leptospirosis, and
A total of 853 lactating Gir × Holstein cows (multi- BVD. Within each operation, cows were randomly assigned
parous, n = 611; primiparous, n = 242) originated from 38 to receive (VAC; n = 153) or not (CON = 134) vaccination
operations were assigned to the following fixed-time AI against IBR, BVD, and leptospirosis (5 mL im of CattleMas-
protocol (FTAI): treatment of estradiol benzoate (2 mg im ter 4+L5, Pfizer Animal Health,) 30 d prior to (d −41) and
of Estrogin; Farmavet, São Paulo, Brazil) and insertion of a at the beginning (d −11) of the FTAI protocol. Hence, the
intravaginal progesterone releasing device (CIDR; contain- objective of this experiment was to evaluate the effects of
ing 1.9 g of progesterone; Pfizer Animal Health) on d −11, vaccination against IBR, leptospirosis, and BVD, with both
158 M.H.C. Pereira et al. / Animal Reproduction Science 137 (2013) 156–162
doses administered prior to FTAI, on pregnancy rates and IBR, BVD, and leptospirosis (5 mL im of CattleMaster 4+L5,
pregnancy losses in dairy operations that did not have a Pfizer Animal Health,) at the beginning (d −11) of the FTAI
history of vaccinating the cowherd. protocol. Hence, the objective of this experiment was to
On d −11, cows were (average ± SEM) 143 ± 4 DIM, evaluate the effects of revaccination against IBR, leptospi-
producing 21.2 ± 0.4 kg of milk, and BCS of 2.94 ± 0.02 rosis, and BVD, administered prior to FTAI, on pregnancy
(Wildman et al., 1982). Pregnancy status and incidence of rates and pregnancy losses in dairy operations that had a
pregnancy losses were assessed as in experiment 1. Blood history of vaccinating the cowherd.
samples were collected from a subsample of CON cows On d −11, cows were (average ± SEM) 149 ± 4 DIM,
from 10 operations (n = 28) on d −11 for determination of producing 24.5 ± 0.3 kg of milk, and BCS of 2.95 ± 0.02
serological profile for BoHV-1, BVDV, and Leptospira spp. (Wildman et al., 1982). Pregnancy status and incidence
infections. These samples were randomly collected from of pregnancy losses were assessed as in experiment 1.
an average of 2 females per operation, being 1 multiparous Blood samples were collected from a subsample of CON
and 1 primiparous cow, processed and analyzed for anti- cows from 11 operations (n = 62) on d −11 for determi-
body detection against BoHV-1, BVDV, and Leptospira spp. nation of the serological profile for BoHV-1, BVDV, and
as described in experiment 1. Leptospira spp. infection. These samples were randomly
collected from an average of 6 females per operation, being
2.4. Experiment 3 4 multiparous and 2 primiparous cows, processed and ana-
lyzed for antibody detection against BoHV-1, BVDV, and
A total of 1680 lactating Holstein cows (multiparous, Leptospira spp. as described in experiment 1.
n = 1160; primiparous, n = 520) originated from 17 oper-
ations were assigned to the experiment. None of these
2.6. Statistical analysis
operations had a history of vaccinating the cowherd against
IBR, leptospirosis, and BVD. Within each operation, non-
For all analyses, significance was set at P ≤ 0.05, and ten-
pregnant cows with DIM greater than 28 d were randomly
dencies were declared if P > 0.05 and ≤0.10. Pregnancy data
assigned to receive (VAC; n = 859) or not (CON = 821) vac-
were analyzed using the GLIMMIX procedure of SAS (SAS
cination against IBR, BVD, and leptospirosis (5 mL im
Institute Inc., Cary, NC, USA) and Satterthwaite approxi-
of CattleMaster 4+L5, Pfizer Animal Health,), with doses
mation to determine the denominator degrees of freedom
administered 14 d apart. Within 15–135 d after the sec-
for tests of fixed effects. The model statement for preg-
ond dose of vaccine administration, cows were artificially
nancy rates and losses contained the effects of vaccination
inseminated 12 h upon visual estrus detection or assigned
treatment, operation, parity, AI technique (experiment 3
to the FTAI protocol described in experiment 1. Hence, the
only; AI or fixed-time AI) and all resultant interactions,
objective of this experiment was to evaluate the effects of
as well as BCS, DIM, and milk production as independent
vaccination against IBR, leptospirosis, and BVD, with both
covariates. However, covariates were removed from the
doses administered only 14 d apart but prior to AI, on preg-
model if P > 0.10. Data were analyzed using cow (opera-
nancy rates and pregnancy losses in dairy operations that
tion × parity × vaccination treatment) as random variable
did not have a history of vaccinating the cowherd.
and error term for tests of fixed effects for experiments
At the time of first treatment administration, cows
1, 2, and 4, and cow(operation × parity × vaccination treat-
were (average ± SEM) 188 ± 3 DIM, producing 33.9 ± 0.2 kg
ment × AI technique) for experiment 3. Cow BCS, DIM, and
of milk, and BCS of 2.77 ± 0.01 (Wildman et al., 1982).
milk production were analyzed using the MIXED procedure
Pregnancy status and incidence of pregnancy losses were
of SAS (SAS Institute Inc.), with Satterthwaite approx-
assessed as in experiment 1, but at 30 and 60 d after AI.
imation and the same models described for pregnancy
Blood samples were collected from a subsample of CON
analysis, but without the independent covariates. Results
cows from 14 operations (n = 130) when VAC cows received
are expressed as least square means (adjusted to the appro-
the first vaccine dose, for determination of the serologi-
priate covariates for pregnancy analysis), separated using
cal profile for BoHV-1, BVDV, and Leptospira spp. infection.
LSD, and are reported according to treatment effects if no
These samples were randomly collected from an average
interactions were significant, or according to the highest-
of 10 females per operation, being 7 multiparous and 3
order interaction detected.
primiparous cows, processed and analyzed for antibody
detection against BoHV-1, BVDV, and Leptospira spp. as
described in experiment 1. 3. Results
A total of 820 lactating Gir × Holstein cows (mul- No differences were detected between VAC and CON
tiparous, n = 643; primiparous, n = 177) originated from cows for BCS (P = 0.16), DIM (P = 0.85), and milk production
15 operations were assigned to the same FTAI protocol (P = 0.38; Table 1). Results associated with seroprevalence
described in experiment 1. All these operations already for BoHV-1, BVDV, and Leptospira spp. are shown (Table 2).
adopted three annual vaccinations against leptospiro- Cow BCS was a significant covariate for pregnancy rates
sis and annual vaccination against IBR and BVD. Within on d 30 (P = 0.03) and 71 (P = 0.04). No vaccination treat-
each operation, cows were randomly assigned to receive ment effects were detected for pregnancy rates on d 30
(REVAC; n = 385) or not (CON = 435) revaccination against (P = 0.68) and 71 (P = 0.41), although pregnancy loss was
M.H.C. Pereira et al. / Animal Reproduction Science 137 (2013) 156–162 159
Table 1 for BoHV-1, BVDV, and Leptospira spp. are shown (Table 2).
Body condition score (BCS), days in milk (DIM), and milk production of
Cow BCS was a significant covariate for pregnancy rates on
cows assigned to each experiment.
d 30 (P < 0.01) and d 71 (P = 0.02), whereas DIM was a signif-
Experimenta BCSb DIM Milk production (kg/d) icant covariate for pregnancy loss (P = 0.04). Cows assigned
Experiment 1 to VAC had greater pregnancy rate on d 30 (P = 0.01) and 71
VAC 2.85 117 19.0 (P = 0.03) compared with CON cohorts (Table 3), whereas
CON 2.91 116 19.6 pregnancy loss did not differ (P = 0.41) between treatments
SEM 0.04 5 0.5
(Table 3).
P-value 0.16 0.85 0.38
Experiment 2
3.3. Experiment 3
VAC 2.97 140 20.0
CON 2.95 149 18.6
SEM 0.05 8 0.9 No differences were detected between VAC and CON
P-value 0.75 0.29 0.15 cows for BCS (P = 0.85), DIM (P = 0.83), and milk production
Experiment 3 (P = 0.75; Table 1). Results associated with seroprevalence
VAC 2.81 207 31.7 for BoHV-1, BVDV, and Leptospira spp. are shown (Table 2).
CON 2.80 206 31.8 Cow BCS was a significant covariate for pregnancy rates on
SEM 0.02 4 0.4
d 30 (P = 0.04) and 60 (P = 0.05). Cows assigned to VAC had
P-value 0.85 0.83 0.75
greater pregnancy rate on d 30 (P = 0.02) and 60 (P = 0.01)
Experiment 4 compared with CON cohorts (Table 3), whereas pregnancy
REVAC 2.89 126.3 22.6
loss did not differ (P = 0.43) between treatments (Table 3).
CON 2.98 159.0 21.3
SEM 0.04 9.4 0.7
P-value 0.04 <0.01 0.05 3.4. Experiment 4
a
In experiment 1, cows received (VAC) or not (CON) vaccination against
IBR, BVD, and leptospirosis on d −11 and d 30 relative to fixed-time AI Cows assigned to REVAC had reduced DIM (P < 0.01) and
(d 0). In experiment 2, cows received VAC or CON on d −41 and d −11 BCS (P = 0.04), and greater milk production (P = 0.05) com-
relative to fixed-time AI (d 0). In experiment 3, cows with DIM greater than
pared with CON cows (Table 1). Results associated with
28 d received VAC or CON, with doses administered 14 d apart and within
at least 15 d prior to AI. In experiment 4, cows received revaccination
seroprevalence for BoHV-1, BVDV, and Leptospira spp. are
(REVAC) or not (CON) against IBR, BVD, and leptospirosis on d −11 relative shown (Table 2). Cow BCS, DIM, and milk production were
to fixed-time AI (d 0). In experiments 1, 2, and 4, variables were recorded significant covariates (P < 0.10) for all pregnancy analysis.
on d −11. In experiment 3, variables were assessed at the time of first No vaccination treatment effects were detected for preg-
treatment administration.
b nancy rate on d 30 (P = 0.99) and 71 (P = 0.90), or pregnancy
As described by Wildman et al. (1982).
loss (P = 0.54; Table 3).
reduced (P = 0.03) in VAC cows compared with CON cohorts
loss (Table 3). 4. Discussion
Table 2
Presence (% of total samples) of antibodies against bovine herpesvirus-1 (BoHV-1), bovine viral diarrhea virus (BVDV), and Leptospira spp. within each
experiment.a,b
Pathogen Titer Experiment 1 (n = 84) Experiment 2 (n = 28) Experiment 3 (n = 130) Experiment 4 (n = 62)
Negative (<8) 0 0 4 2
8–64 27.4 25.0 44 20
BoHV-1
≥64 72.6 75.0 52 78
Positive (≥8) 100.0 100.0 96 98
Pilz, D., Alfieri, A.F., Alfieri, A.A., 2005. Comparison of different protocols Sutton, M.L., 1980. Rapid onset of immunity in cattle after intramus-
for the bovine viral diarrhea virus detection by RT-PCR in pools of cular injection of a modified-live-virus IBR vaccine. Vet. Med. 75,
whole blood and blood serum artificially contaminate. Semin. Cienc. 1447–1456.
Agrar. 26, 219–228. Takiuchi, E., Alfieri, A.F., Alfieri, A.A., 2001. Bovine herpesvirus type 1:
Rebhun, W.C., 1995. Diseases of Dairy Cattle. Williams and Wilkins, Media, infection and diagnosis methods. Semin. Cienc. Agrar. 22, 203–209.
PA, USA. Vogel, F.S.F., Flores, E.F., Weiben, R., Mayers, S.V., Quadros, V.L., Oldonis, I.,
Ryu, E., 1970. Rapid microscopic agglutination test for Leptospira without 2002. Magnitude duration and specificity of the serological response
non-specific reaction. Bull. Off. Int. Epizoot. 73, 49–58. in cattle vaccinated against bovine viral diarrhea virus (BVDV). Ciênc.
Santos, J.E.P., Thatcher, W.W., Chebel, R.C., Cerri, R.L.A., Galvao, K.N., Rural 32, 83–89.
2004. The effect of embryonic death rates in cattle on the effi- Wildman, E.E., Jones, G.M., Wagner, P.E., Boman, R.L., Troutt, H.F., Lesch,
cacy of estrus synchronization programs. Anim. Reprod. Sci. 82, T.N., 1982. A dairy cow body condition scoring system and its relation-
513–535. ship to selected production characteristics. J. Dairy Sci. 65, 495–501.