Twinning in dairy cattle: A study of risk factors and effects

Twin calvings on Dutch dairy farms were studied using data collected through a routine herd health and production management program. Over a period of 6.5 yr, 381 twin calvings (762 calves) of a total of 11,951 calvings were recorded on 33 farms. The 381 twin calvings were compared with a referent group of 1,536 calvings that was randomly selected from the orginal 11,951 calvings. Risk factors for twin calvings were parity, season, herd and previous twinning. High cumulative milk production was associated with cows conceiving twins. Treatment for infertility in the preceding lactation was an equivocal risk factor. Twin calvings increased the risk of abortion, retained placenta, fertility-related treatment, as well as the number of days open and the number of inseminations per conception. Although twin calvings resulted in more calves born per dam, the number of available replacement heifers did not increase. After twin calving, cows produced slightly less milk and were culled more often than single calving cows. To adapt appropriate management practices for cows carrying twins, an early and reliable method for detecting twin pregnancies is needed.     

Induction of twinning in Holstein and Japanese black cows by ipsilateral frozen embryo transfer

Induction of twinning by ipsilateral nonsurgical transfer of two frozen-thawed Japanese Black bovine embryos to each of 20 Holstein and 26 Japanese Black cows, that had been kept under a stable in private farm conditions, was examined. The cows were monitored every 20 days from Day 25 to Day 65 of gestation for pregnancy and fetus survival (estrus is Day 0). Seventy-five per cent (15 of 20), 65.0% (13 of 20) or 60.0% (12 of 20) and 61.5% (16 of 26), 53.8% (14 of 26) or 50.0% (13 of 26) of Japanese Black cows were diagnosed pregnant at 25, 45 and 65 days after transfer by ultrasonic echography. Embryonic losses were observed between Days 25 and 65 in 29.2% (7) Holstein and 31.8% (7) Japanese Black cows. The twin pregnancy rate in Holstein and Japanese Black cows decreased with time; 60.0% (9 of 15) vs. 37.5% (6 of 16) at Day 25; 53.8% (7 of 13) vs. 28.6% (4 of 14) at Day 45 and 41.7% (5 of 12) vs. 15.4% (2 of 13) at Day 65. At calving, Holstein cows produced five sets of twins and seven single calves, and Japanese Black cows two sets of twins and 11 single calves. The twinning rate in Holstein cows was higher (P < 0.05) than that in Japanese Black cows, 41.7% (5 of 12) vs. 15.4% (2 of 13). The calf birth weight in Holsteins was heavier (P < 0.05) than that in Japanese Black dams (24.5 kg, 33.6 kg vs. 19.3 kg, 25.5 kg for twin and single calves). The placental weight in Holstein dams calving twins was heavier than that in Holstein dams calving a single calf or in Japanese Black dams calving either twins or a single calf (6.6 kg vs. 3.5 kg, 4.6 kg or 2.8 kg). The number of placentome in Holstein dams calving twins was also higher (P < 0.05) than that in Holstein dams calving a single calf or Japanese Black dams (103.5 vs. 41.8, 67.9, 33.0). The number of placentome was approximately double in dams calving twins than that of dams calving a single calf.     

Postpartum fertility in beef cattle producing twins

Postpartum fertility was measured in 42 plur iparous Hereford cows and first calf Hereford heifers that calved after embryo transfer to induce twins. Dams were exposed to a Hereford bull from 4: 00 P.M. to 8:00 A.M. each day from 60 days postpartum until pregnancy was confirmed or calves were weaned at 180 days of age. Days open ($\text{X}\text{± SE}$) for dams that produced single and twin calves were 84.1 ± 4.6 (n = 12) and 94.9 ± 6.2 (n = 30), respectively. Corresponding values for dams that nursed one calf, including six females that lost one calf of a twin set at birth, and dams that nursed twins were 89.3 ± 6.4 (n = 18) and 93.4 ± 8.5 (n = 24). No significant differences were observed due to calving or suckling twin calves. Heifers that calved twins had a shorter mean interval to conception than cows that calved twins. These results are interpreted to mean that with proper management during the prepartum and postpartum periods, reduced fertility in beef cattle that produce twins need not occur.     

Importance of dam BW change and calf birth weight in double-muscled Belgian Blue cattle and its relationship with parity and calving interval

Factors affecting calving interval (CI) in double-muscled Belgian Blue (DMBB) beef cows were investigated with regard to the BW yield (BWY) of the cow–calf pair, using 834 CI records from 386 females with parities 1 to 6. The effect of parity and CI on BWY was also studied. Cow–calf pair BWY was defined as calf birth weight plus dam BWY per CI. CI (mean±s.e.: 404±1.9 days) was affected by parity, calving season, suckling and calf birth weight/dam weight. Primiparous cows had a shorter CI than cows with three or more calvings (P<0.05), with an intermediate CI for second-calf cows. Spring calvings resulted in a shorter CI than summer and autumn calvings, with intermediate values for winter calvings. Suckling dams had longer CIs than non-suckling dams. There were interactions (P<0.05) between calving season and suckling, and between calving season and mating system. Shortest CIs were observed for spring calvings in case of non-suckling and for summer calvings in case of suckling. Longest CIs were observed for autumn calvings in case of natural service (NS) and for winter calvings in case of artificial insemination (AI). Calf birth weight/dam weight of 6% to 10% resulted in shorter CI than a ratio of <6% (P<0.05). Body condition and mating system (NS v. AI) did not affect CI. Daily cow–calf pair BWY was affected by parity (P<0.001) and CI (P=0.013), with a tendency for an interaction (P=0.094). Daily cow–calf pair BWY did not differ for CIs of <12 to 16 months in primiparous cows and was lowest for a CI of 13 to 15 months in second-calf cows, whereas the effect of CI was more variable in older cows. Dam contribution to cow–calf pair BWY was larger than calf birth weight in first- and second-calf cows, and increased with increasing CI. Dam contribution to cow–calf pair BWY was smaller than calf birth weight in older cows, varying from 0.2 to 1.0 depending on CI. A short CI is advised for DMBB cows because of a larger BWY and more efficient nutrient utilisation.     

Induction of twinning in crossbred heifers by ipsilateral frozen embryo transfer

Induction of twinning by means of nonsurgical ipsilateral transfer of two frozen embryos or of demi-embryos was attempted in 297 virgin crossbred dairy heifers that were kept at the following locations: in stables at our breeding station (Group 1), in a mountain field (Group 2) and at a private farm (Group 3). Of these, 187 heifers (63%) were diagnosed pregnant at 25 d after embryo transfer and 134 heifers (45%) were diagnosed pregnant at 60 d after transfer by ultrasonic echography or by palpation per rectum. At calving, 118 heifers produced 46 sets of twins and 72 single calves, for a total of 164 calves sixteen pregnant heifers aborted fetuses between 2 and 7 mo. The pregnancy rate of Group 1 (39%) was lower than that of Group 2 (48%) or Group 3 (50%). Abortion and mortality rates for Group 3 (8% and 8%, respectively) were significantly lower (P<0.05) than for Group 1 (16% and 12%) or Group 2 (13% and 18%). Twin calves had lower birth weights (P<0.05) than singles, but at 270 to 330 d of age, there was no significant difference between the weights of single and twin calves. Dystocia or difficult delivery was not observed heifers producing twins. Retention of the placenta was observed in twin calving heifers, with placenta retention levels the highest in Group 2 (33%) than followed by Group 1 (14%) and Group 3 (10%). When gestation periods were compared between single and twin calves in Groups 1 and 2, the gestation period of single calves was 5.0 to 7.0 d shorter than that of twin calves. No significant difference was observed between the gestation periods of single and twin births in Group 3.     

Factors affecting secondary sex ratio in Iranian Holsteins

The objective of this study was to evaluate the factors affecting secondary sex ratio (SSR) in Iranian Holsteins. Data of 942,941 Holstein calving events from the Animal Breeding Center of Iran, recorded between January 1996 and December 2007, were used in the analysis. A multivariable logistic regression model was used to model the logit of the probability of a male calf being born. Male births accounted for 49.6% of the total observations. The ratio of males to females varied from 52.5:47.5 in calving year 1996-1999 (odds ratio (OR) = 1.18; P < 0.0001), to 48.5:51.5 in calving year 2004-2007. The greatest occurrence of male births was observed in spring (OR = 1.02; P < 0.0001), and the lowest incidence of male births was for summer or fall calvings. Also, the frequency of male births decreased from parity 1 to parity 4 and beyond (P < 0.0001; OR = 1.11). The greatest number of sires had the SSR equal to 0.5 with a minimum SSR of 32% while the maximum was 97%. Among cows that had a male birth, the chance of delivering a male calf again was 25.5% when cows had delivered a male once (OR = 1.14; P < 0.0001), and 12.7% if a male calf was delivered twice by a cow. This indicated that characteristics peculiar to the dam influence the sex of her offspring and suggests some degree of repeatability of calf sex within cows.     

Abnormal offspring following in vitro production of bovine preimplantation embryos: a field study

Data on 944 calves from 2228 in vitro-produced (IVP) bovine preimplantation embryos were compared with data on 2787 AI calves born in the same herds in 1995. Bovine preimplantation embryos were produced in vitro following ovum pick up (OPU) from donor cows and pregnant heifers in an open nucleus breeding program. After 7 d of in vitro culture on a BRL cell monolayer in the presence of 10% FCS, frozen-thawed expanded blastocysts and fresh morulae to expanded blastocysts were transferred into recipient heifers and cows at 119 contracted farms throughout the Netherlands. The pregnancy rate, as confirmed by palpation per rectum between 90 and 150 d after transfer was 43.5% for both fresh and frozen embryos. Data on IVP and AI calves were registered by the farmers. The percentage of calves with a congenital malformation and the percentage of male calves were related to the total number of calves born. Gestation length, birth weight (measured by a balance), perinatal mortality and ease of calving were analyzed in a subdataset (699 IVP and 2543 AI calves, respectively) by a comparative analysis of variance (ANOVA). The ANOVA model included herd, month of calving, sire nested within AI or IVP, parity and breed of the inseminated cow/embryo recipient, sex of calf, type of calf (AI or IVP) and two-way interactions between type of calf and sex, parity and breed. The percentage of calves with congenital malformations was 3.2% and 0.7% for IVP and AI calves, respectively. An increased incidence of hydro-allantois and abnormal spinal cords and limbs was observed in IVP calves. The percentage of male calves was significantly different between IVP and AI, 55.5% and 48.9%, respectively (Chi-square, 1 degree of freedom, P < 0.05). On the average, IVP calves showed a significant increase of birth weight by 10% (4-5 kg), a 3-d longer gestation period, 2.4% more perinatal mortality and a more difficult calving process compared to AI calves (P < 0.05). From these results it is concluded that calves produced by IVP deviate significantly from calves produced by AI.     

Influence of bull biostimulation, season and parity on resumption of ovarian activity of zebu (Bos indicus) cattle following parturition

A total of 135 postpartum suckled zebu cows were assigned randomly at calving to two treatments: cows exposed to vasectomised bulls (Mature Bull Exposure, MBE) and cows not exposed to bulls (No Bull Exposure, NBE) for a period of 150 days postpartum. This study was conducted to determine the influence of bull biostimulation, season and parity on postpartum reproductive performance of Bos indicus cattle. The trials were conducted in two seasons: cows calving in the dry season and cows calving in the rainy season. Cows with an increase in serum progesterone (P(4)) concentration of >1 ng/ml from the weekly blood samples were used to analyse the number of days from calving to the time of resumption of ovarian activity. The introduction of vasectomised bulls shortened the postpartum anoestrus in cattle following calving. Mean interval from calving to resumption of ovarian activity for the MBE cows was 71.7 days, which was significantly earlier following calving than the NBE cows with a duration of 77.8 days. By 60 to 80 days postpartum, the proportion of cows at resumption of postpartum activity for MBE cows was greater than for the NBE cows.Mean interval from calving to resumption of ovarian activity for cows that calved in the dry season was 71.3 days, which was significantly earlier than for cows that calved in the rainy season (78.6 days). At 60 days postpartum, the proportion of cows at resumption of postpartum ovarin activity for cows that calved in the dry season was greater than the cows that calved in the rainy season. Mean interval from parturition to resumption of ovarian activity for cows with three to five calvings was 65.1 days, which was significantly earlier than the value of 71. 2 days for cows with one to two calvings. By 60 to 80 days postpartum, the proportion of cows at onset of ovarian activity for cows with three to five calvings was greater than those cows with one to two calvings. Cows that calved in the dry season completed uterine involution by 24.4 days, which was significantly shorter than the duration of 26.5 days for cows that calved in the rainy season. Cows with three to five calvings completed uterine involution earlier than those with one to two calvings. It is concluded that bull-cow biostimulation influences reproductive activity in the cow possibly through olfactory cues (pheromones).     

Management of Calving in Norwegian Cubicle-Housed Dairy Herds

Sixty of the 65 dairy farms with cubicle houses in the Norwegian county of Oppland were included in a field study of the management of calving in 1990. The farmers recorded the location of the cow when giving birth, farmer presence and whether assistance was given during calving, occurrence of suckling, and time after birth when cow and calf were separated. Such data were recorded for a total of 1125 calvings. About 10% occurred on pasture, while 78% of the remaining calvings took place in the cubicle-equipped section. Thirteen percent calved in a calving pen, the remaining cows being tethered at the time of calving. Thirty-two percent of the calvings took place in houses lacking a calving pen altogether. Farmers were present during 41% of the calvings. Suckling most frequently occurred after pasture calvings, and was least frequent after calvings within the cubicle-equipped section of the cowhouse. Injuries to the calf caused by trampling or contact with fittings etc. were rare, and no more common in association with calving in the cubicle-equipped section than with calving taking place with the cow isolated from the rest of the herd. All calves were removed from their dams within 24 h after birth.     

Mortality in Holstein-Friesian calves and replacement heifers, in relation to body weight and IGF-I concentration, on 19 farms in England

The incidence of mortality and culling in Holstein-Friesian heifers from birth through first calving was determined on 19 dairy farms selected from across southern England. The outcome of 1097 calvings was determined. Size (BW, heart girth, crown-rump length and height at withers) and insulin-like growth factor-I concentration of live heifer calves were measured at a mean age of 26 ± 0.7 days (n = 506). Associations between the heifer-level variables and mortality were determined using clustered binary logistic regression. Perinatal mortality (stillbirths and mortality within the first 24 h of birth) of male and female calves was 7.9%. This figure was significantly higher in cases where calving assistance was required (19.1% v. 5.6%, P < 0.001) and in twin births (18.5% v. 7.0%, P < 0.05), and was lower in pluriparous v. primiparous dams (5.6% v. 12.1%, P < 0.01). On average, 6.8% of heifers died or were culled between 1 day and 6 months of age. Low BW at 1 month was associated with reduced subsequent survival up to 6 months. Between 6 months and first calving, a further 7.7% of heifers either died (42%) or were culled (58%); accidents and infectious disease accounted for the majority of calf deaths between 6 and 15 months, whereas infertility (16/450 animals served, 3.5%) was the main reason for culling following the start of the first breeding period. In total, 11 heifers (2.2%) were culled as freemartins; eight at birth and three around service. Overall, 14.5% of liveborn potential replacement heifers died or were culled before first calving.     

Danish Holsteins Favor Bull Offspring: Biased Milk Production as a Function of Fetal Sex,and Calving Difficulty

In a previous study from 2014 it was found that US Holstein cows that gave birth to heifer calves produced more milk than cows having bull calves. We wanted to assess whether this is also true for Danish cattle. Data from 578 Danish Holstein herds were analysed with a mixed effect model and contrary to the findings in the US, we found that cows produced higher volumes of milk if they had a bull calf compared to a heifer calf. We found a significantly higher milk production of 0.28% in the first lactation period for cows giving birth to a bull calf, compared to a heifer calf. This difference was even higher when cows gave birth to another bull calf, so having two bull calves resulted in a difference of 0.52% in milk production compared to any other combination of sex of the offspring. Furthermore, we found that farmer assisted calvings were associated with a higher milk yield. Cows with no farmer assistance or with veterinary assistance during the most recent calving produced less milk. There were also indications that dams would favor a bull fetus by decreasing milk production during the second pregnancy if the calf born in the first parity was a heifer. We hypothesize that size of calves is a confounding factor for milk production. However, calving weight was not available in the present data set to test this hypothesis.     

Artificial induction of twinning in cattle by means of supplemental embryo transfer

The induction of twins by surgical and nonsurgical transfer was performed 6 - 8 days after Artificial Insemination (A.I.) in a total of 142 cows. Of the 47 heifers, which received surgically transferred embryos, 32 (68%) became pregnant. Of the 95 cows, which received nonsurgically transferred embryos, 58 (61,6%) became pregnant. A total of 90 (63.5%) of the recipient animals became pregnant. Up to date 64 recipients have calved; twins have resulted in 40 and 48,2% resp, from surgical and nonsurgical inductions. Out of a total of 64 calvings 29 (45,3%) were twins, and of these 38% were males, 27% were females, and 34% were mixed births. Female - calves resulted in freemartinism. There was no increase in the incidence of either retained placenta or dystocia. The induction of twinning by means of embryo transfer can be regarded as a useful method for the increase in the reproductive performance cattle.     

Prevalence of, and risk factors associated with, perinatal calf mortality in pasture-based Holstein-Friesian cows

Recent publications indicate that the prevalence of perinatal mortality has increased in some dairy industries and an increased proportion of this loss is not associated with the traditional risk factors for perinatal mortality. The objectives of this study were to establish the prevalence of perinatal mortality (calf death within 24 h of calving) in Irish dairy herds and to determine the current significance of putative risk factors in pasture-based management systems. A total of 182 026 records of full-term calvings from Holstein-Friesian dams served by artificial insemination (AI) sires of seven breeds in herds of 20 calvings or more per year were available from the Irish national breeding database over 4 years (2002 to 2005). The prevalence of perinatal mortality was 4.29% (7.7% in primiparae and 3.5% in pluriparae). The likelihood of perinatal mortality increased between 2002 and 2005 and was greatest in June and in winter. There was an interaction (P < 0.001) between the effect of calving assistance and parity with the effect of dystocia on perinatal mortality being greater in primiparae. The odds of perinatal mortality were greater in male (OR = 1.12; P < 0.001) and in twin calves (OR = 5.70-13.36; P < 0.001) and in dams that had perinatal mortality at the previous calving (OR = 4.21; P < 0.001). The logit of the probability of perinatal mortality increased by 0.099 per unit increase in sire predicted transmitting ability (PTA) for direct perinatal mortality. The probability of perinatal mortality increased at an increasing rate in primiparae as animals calved at a younger age relative to the median age at first calving. The only herd-level factor examined, herd size did not affect the odds of perinatal mortality. These data indicate that the prevalence of perinatal mortality in this cattle population is similar to that in other pasture-based dairy systems worldwide. The putative exposures and attributes traditionally associated with perinatal mortality were associated with perinatal mortality in this pasture-based dairy cow population. The practical implication of these results is that as many of the significant risk factors are largely not under management control (year of calving, month of calving, twin calving, primiparity, previous perinatal mortality and foetal gender), herd owners must focus on the significant determinants under their control (age at first calving, sire genetic merit for direct perinatal mortality and both the extent of calving supervision and the degree of assistance), in order to reduce the prevalence of perinatal mortality and improve perinatal welfare.     

The fertility of autumn calving suckler beef cows is increased by the addition of prostaglandin to progesterone and eCG estrus synchronization treatment

The objective of this study was to determine the efficacy of PGF2 alpha treatment on pregnancy and calving rates in autumn-calving suckler beef cows synchronized with progesterone and eCG. The population studied consisted of 124 Charolais and 130 Limousin cows in 13 and 12 beef herds, respectively. In each herd, pairs of cows were formed according to parity, body condition score and calving difficulty. Group 1 received a progesterone releasing intravaginal device (PRID) for 12 d with a capsule containing 10 mg estradiol benzoate at implant insertion and 500 IU eCG at PRID removal (Day 0). Group 2 received the same treatment plus 25 mg i.m. dinoprost at Day -2. Each cow was artificially inseminated 56 h after PRID removal (Day 3). Plasma progesterone concentrations were measured to determine cyclicity prior to treatment in samples take on Days -22 and -12, to confirm the occurrence of ovulation (Day 13) and to determine the early pregnancy rate (Day 26). Serum pregnancy-specific protein B (PSPB) concentrations were determined to assess pregnancy rate at Day 39. The effects of variation factors on pregnancy and calving rates after treatment were studied using logistic mixed models and a Cox model, respectively. There were no significant differences between groups or breeds for the rate of cyclicity before treatment nor for ovulation rate (means, 74.1 and 95.7%, respectively). Cyclicity was, however, influenced by individual factors such as body condition score (OR = 3.36, P = 0.001), parity (OR = 5.4, P = 0.001) and herd factors such as stocking rate (OR = 5.62, P = 0.001). The use of a prostaglandin injection increased pregnancy rate at Day 26 (71.7 vs 56.7%, P = 0.01) and at 39 d (67.7 vs 54.3%, P = 0.02) and the calving rate at induced estrus (64.5 vs 48.5%, P = 0.01). We observed 9 twin calvings (5.6%) which occurred in cyclic cows only before treatment. Cows in Group 2 had a 1.5 greater chance of calving before 300 d following the first AI than cows in Group 1 (P = 0.03). In conclusion, the addition of PGF2 alpha injection, 48 h before PRID removal, increased reproductive efficiency in autumn-calving Charolais and Limousin suckler beef cows compared to a classical estrus synchronization treatment using a PRID + eCG.     

Pregnancies, calves and calf viability after transfer of in vitro produced bovine embryos

Pregnancy, parturition and calf survival following the transfer of embryos produced in vitro were monitored. A total of 44 blastocysts was transferred in pairs to 1 uterine horn ipsilateral to the corpus luteum (CL) of 22 synchronized heifers. At Day 42 of development 14 recipients (64%) were pregnant; the calving rate was also 64%. The twinning rate was 9/14 at Day 42 and 7/14 at birth, for an overall fetal mortality rate of 9%. The average gestation length was 281 and 275 d for single and twin pregnancies, respectively. Blood samples from recipients were collected for determination of bovine pregnancy associated glycoprotein (bPAG) from 2 wk after transfer and throughout the pregnancy. During the first trimester of pregnancy, the bPAG concentration was significantly higher in twin than in single bearing heifers, and the perinatal increase in bPAG was correlated positively with the total weight of the fetus(es). The percentage of male calves was 43%. The birth weight of twin individuals was 25 +/- 1 kg, which was 78% of the birthweight of the singletons (32 +/- 2 kg). One singleton calf was oversized, weighing 58 kg (80% more than the median weight of the other singletons). Stillbirths occurred in 21% of the twins, butin none of the singletons. Calf mortality during the first 14 d was higher for twins (4/11) than for singletons (1/7) due to infections and cerebellar hypoplasia. Karyotyping the calves detected no cytogenetically recognizable abnormalities. All calves were negative for BVD virus and IBR antibodies. The results of this study showed that although the incidence of fetal loss was low, there was an unacceptable high perinatal mortality of the calves. Thus it is likely that the blood supply through the placenta of animals pregnant with twins was impaired or it is possible that these fetuses and calves had increased stress susceptibility caused by the in vitro conditions. Furthermore, the birth of 1 oversized calf, 2 calves with cerebellar hypoplasia and 5 calves succumbing to infections seems to indicate that a proportion of in vitro produced calves may suffer from factors inherent in the in vitro production system.     

The use of crossbreeding with beef bulls in dairy herds: effects on calving difficulty and gestation length

This study was designed to analyse the evolution in the use of beef bull semen for dairy cattle insemination and, mainly, to assess calving difficulty, gestation length and proportion of stillbirths after breeding pure Holsteins or crossbreeding. Data were collected during 2004 to 2011 for 552 571 Holstein calvings (457 070 Holstein × Holstein, 43 384 Holstein × Limousine, 32 174 Holstein × Belgian Blue and 19 943 Holstein × Galician Blonde). The highest calving difficulty, compared with pure Holsteins was for crosses with Belgian Blue followed by Limousine and Galician Blonde. The Holstein × Limousine and Holstein × Galician Blonde crossbred calves had significantly longer gestation lengths than Holstein × Holstein and Holstein × Belgian Blue calves. Between the latter two, pure Holstein had the shortest gestation length. Calving difficulty and gestation length decreased as the age of the dam advanced. The most difficult calvings were observed in twin calvings, followed by the calvings of male calves and female calves. The gestations leading to the birth of male calves were longer than those leading to female calves and twin calves. Stillbirths were not related to the breed used for mating. Through examining these parameters, sire breed should be considered when selecting a beef breed for the insemination of milk-producing dams.     

Normal calves from transfer of biopsied, sexed and vitrified IVP bovine embryos

Data on biopsied, sexed and cryopreserved in vitro produced (IVP) bovine embryos, and their in vivo developmental competence are very limited. Two preliminary studies were conducted before the primary study. In Experiment 1, post-thaw in vitro developmental competence of biopsied and vitrified IVP embryos was evaluated using re-expansion as an endpoint. In Experiment 2, the pregnancy rates of biopsied fresh, frozen or vitrified embryos following single embryo transfer were compared. Since vitrified embryos resulted in a higher pregnancy rate than frozen-thawed embryos, in the primary study (Experiment 3), all IVP embryos were vitrified following biopsy and sexing (by DNA fingerprinting). In Experiment 3, we compared pregnancy initiation and calving results of heifers in the following treatments: 1) artificial insemination (AI); 2) AI plus contralateral transfer of a single embryo (AI + SET); 3) ipsilateral transfer of single embryo (SET); or 4) bilateral transfer of two embryos (DET). Birth weights, gestation lengths and dystocia scores were recorded. In Experiment 1, post-thaw re-expansion rate of biopsied and vitrified embryos was 85% (70/82). In Experiment 2, pregnancy rates (90 d) were 44% (7/16), 23% (3/13), and 50% (7/14) for vitrified, frozen and fresh embryos, respectively (P < 0.10). In Experiment 3, pregnancy rates of AI and SET were 65% (20/31) and 40% (16/40), respectively (P < 0.05). The pregnancy rate of AI + SET was 75% (27/36) with 11 carrying twins, and the pregnancy rate of DET was 72% (26/36) with 10 carrying twins. All AI fetuses were carried to term, but only half the SET fetuses were carried to term. Similar calving rates were observed in the AI + SET and DET groups, 76 and 70%, respectively, of those pregnant at Day 40. Mean birth weight, dystocia score and gestation length of AI calves were not different from those of SET calves. Mean birth weight and dystocia score of single-born calves were greater than those of twin born calves (P < 0.05). These data demonstrate that biopsied IVP bovine embryos can be successfully cryopreserved by vitrification and following post-thaw embryo transfer, acceptable rates of offspring with normal birth weights can be obtained without major calving difficulties.     

Free radicals, lipid peroxidation and the antioxidant system in the blood of cows and newborn calves around calving

The oxidative stress of birth in cattle (Bos taurus) was evaluated by measuring steady state concentration of free radicals in whole blood, rate of lipid peroxidation and activity of antioxidant enzymes in erythrocytes, antioxidant capacity of blood plasma in 14 calves at birth and four times after birth until 3 weeks of age and also in their mothers at calving. The same parameters were also measured in 58 dairy cows before calving, at parturition and after calving. Free radical concentration in the blood of newborn calves was higher than in cows confirming that birth means oxidative stress for calves. Red blood cell malondialdehyde in calves was the highest at birth and following the first solid feed intake at the third week. Superoxide dismutase activity increased in calves during the first three weeks of life. Ferric reducing ability of plasma was higher in calves at birth than in cows and decreased thereafter. Higher superoxide dismutase activity in red blood cells and lower ferric reducing ability of plasma in dairy cows was found at calving compared to the average of all pre- and post-calving results. We conclude that the blood of newborn calves is well prepared to deal with the oxidative stress of birth, and that such a stress is present even when some fingerprint markers of redox imbalance show no apparent alterations. Stress of calving has minor effects on the antioxidant system of cows.     

Analysis of body measurements of newborn purebred Belgian Blue calves

At calving, purebred animals of the Belgian Blue (BB) breed are compromised by the incompatibility in size and shape of the dam and her calf, resulting in a very high incidence of dystocia problems. To clarify which body parts of the calf are of decisive importance to allow natural delivery and to investigate both the mean value as well as the variation among these body sizes within this breed (variation being an important condition for selection), measurements of nine body parts (body weight at birth (BW), body length (BL), length of the head (LH), shoulder width (SW), hip width (HW), heart girth (HG), withers height (WH) and the circumference of the fetlock of both the front (CFF) and the hind leg (CFH)) were assessed in 147 newborn purebred BB calves on 17 farms. Simple and partial correlations were assessed and we examined whether environmental factors (gender of the calf, parity of the cow, type of calving, season of birth and time of measurement after birth) were significantly associated with these specific calf measurements. The mean BW was 49.2 ± 7.1 kg. The average BL was 56.4 ± 4.5 cm and the mean LH was 24.4 ± 2.3 cm. Measurements obtained for SW and HW were 22.4 ± 2.2 and 22.9 ± 2.1 cm, respectively, whereas the mean WH was 71.1 ± 4.7 cm. Measurements of circumferences revealed a CFF of 17.9 ± 1.1 cm, a CFH of 18.0 ± 1.0 cm and a mean HG of 78.0 ± 5.4 cm. Partial correlations of the BW with eight body measurements were significant (P < 0.01) and ranged between 0.17 and 0.85; 0.42 and 0.88; and 0.24 and 0.88 when corrected for gender, parity and type of calving, respectively. BL (P < 0.01) and the CFF and CFH (P < 0.001) are larger in bull calves than in heifer calves. Calves born through caesarean section had broader SW (P < 0.01) and HW (P < 0.01) when compared with calves born after natural calving (defined as born per vaginam without assistance or with slight traction). Sizes of calves born out of multiparous cows were generally larger than of calves born out of heifers (SW: P < 0.001; HW: P < 0.05). As SW and HW are the broadest points of a BB calf, they are both candidates for being the limiting measures for calving ease, but the difference between HW and SW for the total data set was not different from zero (P > 0.05). In contrast to male calves in which no significant difference (between HW and SW) could be found, female calves show the difference between HW and SW that was significantly different from zero (P < 0.001); thus, in female calves, the HW is the most limiting factor of the calf's body. The significant variation in some body measures between the calves and the strong correlation within these sizes raises the possibility of selection towards smaller calves aiming to limit the dystocia problem in the BB breed. Furthermore, on the basis of our results, we were able to build equations for the farmer to use at the moment of calving containing the LH, the CF and the calf's gender to estimate SW and HW, the limiting body parts of the calf to be born naturally. Together with the knowledge of the pelvic size of the dam, this information gives the obstetrician or the farmer a more accurate prediction of the probability of natural calving at parturition.     

Production efficiency of Japanese black calves by transfer of bovine embryos produced in vitro

The objective of this study was to examine the production efficiency of Japanese Black beef calves after transfer of bovine embryos derived from an in vitro procedure. In vitro-produced (IVP) embryos were obtained from in vitro maturation and fertilization and in vitro development by co-culture with cumulus cells until 7 or 8 days after insemination. In vivo-developed (IVD) embryos from superovulated Japanese Black heifers and cows 7 days after artificial insemination were used as a control group. Bovine embryos were transferred nonsurgically to recipient cows on Day 7 +/- 1 of the estrous cycle. Pregnancy was diagnosed by palpation per rectum at Day 60 to 70 after estrus. Pregnancy, abortion, perinatal accident and birth rates were examined according to the origin of embryos (IVP or IVD), the number of transferred embryos (single or twin) and the storage status (fresh or frozen-thawed). In Experiment 1, production efficiency by twin transfer of fresh IVP embryos was examined. Higher pregnancy rates (52 1% vs 42 9%, P < 0.05) and birth rates (47.0% vs. 33.0%, P < 0.05) were obtained by twin transfer than by single transfer of fresh IVP embryos. Thus, the twin transfer of fresh IVP embryos was effective for production of calves, although the birth rates for single and twin transfers of fresh IVD embryos were still higher (55.5% and 76.1%, P < 0.05). But the abortion and perinatal accident rates for twin transfer of fresh IVP embryos were also significantly greater than those for single and twin transfer of fresh IVD embryos (P < 0.05). In Experiment 2, production efficiency by twin transfer of frozen-thawed IVP embryos was examined. Either single or twin transfer of frozen-thawed IVP embryos resulted in a similar pregnancy rate (41.3% vs. 46.7%, P > 0.05) and birth rate (34.1% vs. 41.1%, P>0.05). Thus, in combination with frozen-thawed IVP embryos, the twin transfer did not enhance production efficiency. In conclusion, Japanese Black beef calves could effectively produce calves by twin transfer to Holstein recipients when using fresh IVP embryos, and by single transfer when using frozen-thawed IVP embryos.