Extraction forces in bovine obstetrics: An in vitro study investigating alternate and simultaneous traction modes

Whether extraction of a calf in longitudinal anterior presentation should be carried out by simultaneous or alternate traction on the forelimbs remains controversial. Because most recommendations are based on empirical observations rather than on scientific studies, the aim of this study was to develop an in vitro model to objectively compare the forces occurring during alternate and simultaneous traction. In a biomechanical in vitro model, 12 dead Holstein-Friesian (Bos taurus) calves were pulled through the prepared pelvic specimen of a cow at a controlled speed using two electric motors. Traction was applied simultaneously (ST) to both legs or alternately (AT) to one leg at a time to advance the calf 5 cm (AT 5) or 10 cm (AT 10). Forces on each limb were measured digitally using load cells. In all cases, two peaks of maximum force occurred during the extraction of the cranial part of the body. The first peak was observed when the elbows were pulled into the pelvis, and the second peak occurred when the chest emerged from the pelvis. Up to and including entry of the elbows into the pelvis, the maximum force on a single limb (341 ± 106 N) was lowest (P < 0.01) using AT10. The maximum traction forces acting on a single limb using AT5 (411 ± 86 N) and ST (431 ± 127 N) did not differ (P > 0.05). During extraction of the thorax, the maximum force acting on a single limb was lower (P < 0.0001) using ST (352 ± 98 N) compared with AT5 (432 ± 79 N) and AT10 (547 ± 115 N). Based on these findings, alternate-limb traction, 10 cm at a time, should be used until both elbows have entered the pelvis. Simultaneous traction should then be applied to both forelimbs to complete extraction of the chest.     

Mode of delivery is associated with different hematological profiles in the newborn calf

Several studies on babies have shown that the type of delivery can influence the hematological and immune status of the newborn. In bovine medicine, some authors reported the hematological pattern of the newborn calf, but never related it with the calving process or other perinatal factors. The purpose of the present study was to evaluate the hematological profile in newborn calves in relation to the type of delivery. A total of 41 healthy calves were enrolled; 16 Friesian calves which were born by vaginal delivery without assistance (VD), and 25 Belgian Blue calves that were born by elective Caesarean section (CS). As soon as the calves were born, a complete clinical examination was performed to verify viability and maturity. At 10 min after birth, 2 mL venous blood was collected to perform the blood gas and acid-base evaluation. Blood samples were subsequently collected from the jugular vein within 30 min after birth, and at 1, 2, 3, 7, and 14 days of age. An automatic analyzer was used to determine hemoglobin concentration (Hb), hematocrit (Ht), and red and white blood cell counts, while differential leukocyte count was performed microscopically. Statistical analysis was applied to assess differences between the groups and within the group for all parameters between each sampling time (P ≤ 0.05). All the calves were born alive, viable, and mature. There were no acidotic calves, but statistical analysis revealed many differences, as higher pH, base excess (BE) (P ≤ 0.05), PO₂ (P < 0.001), and sO₂ (P < 0.0001) in the VD group. Levels of hemoglobin concentration, hematocrit, and red blood cell number were constantly higher in CS calves (P < 0.001). In comparison with the VD calves, white blood cell and neutrophil absolute number were higher at birth and at 14 days of age in the CS group (P < 0.001 and P ≤ 0.05). The mode of delivery, therefore, seems to have an influence on the oxygenation levels and on the hematological and nonspecific immunity profile of the newborn calf.     

Application of computed tomography for the evaluation of obstetrically relevant measurements in German Holstein-Friesian calves

The aim of this study was to measure obstetrically relevant dimensions in calves manually and via computed tomography and to investigate their relationship with the dimensions of fetal body parts that are accessible to the obstetrician during early stages of vaginal delivery. Twenty Holstein-Friesian stillborn calves (Bos taurus) weighing 41.1 ± 3.7 kg (33.6-46.5 kg) were examined and the maximum height (H), width (W), circumference (C) and cross-sectional area (A) of their body was determined. The largest (P < 0.05) one-dimensional variable was the height of the thorax in the region of cranial sternum (H-Thorax; 29.3 ± 1.3 cm), and the largest (P < 0.05) two-dimensional variables were the cross-sectional area of the shoulder region at greater tubercles of the humeri (A-Shoulder; 307 ± 27 cm²) and of the thorax in the region of cranial sternum (A-Thorax; 306 ± 25 cm²). The dimensions of the front legs, which included the circumference of the canon bone and width of the fetlock joint, did not correlate (P > 0.05) with H-Thorax, A-Shoulder and A-Thorax. There were moderate significant correlations between the perpendicular height of the head (H-Head) and A-Thorax (r = 0.65, P < 0.05) and between the circumference of the head (C-Head) and A-Shoulder (r = 0.64, P < 0.05) and A-Thorax (r = 0.52, P < 0.05), but other relationships (P > 0.05) between the dimensions at the level of the head and H-Thorax, A-Shoulder and A-Thorax were not significant (P > 0.05). The results of this study show that the shoulders and thorax are the obstetrically most relevant calf dimensions, but the size of these variables cannot be reliably predicted by evaluating the dimensions of the forelimbs and head of calves during delivery.     

Ultrasonographic fetal parameters and neonatal survival in somatic cell nuclear transfer–derived beef calves

The objectives of this study were to identify prognostic indicators of calf survival in SCNT-derived beef calves. Ultrasonographic parameters of fetal well-being and development, maternal clinical parameters, and neonatal parameters were evaluated as predictors of calf survival in cows carrying SCNT-derived beef fetuses (n = 38). Calf survival was 61.5% (88.2% female and 40.9% male calves; P = 0.0026). Cow respiratory rate and cow temperature were significantly greater in the nonsurviving (NS) group 1 week prepartum. In surviving (S) calves, fetal heart rate (FHR) decreased during the last 2 weeks of gestation (P < 0.01). However, this final deceleration was not observed in NS calves, resulting in higher FHRs in this group (P < 0.0001). Fetal movement and fluid scores did not differ with calf classification. Mean amniotic fluid depth was smaller in S (5.5 ± 0.7 cm) than NS (8.7 ± 1.4 cm) calves (P = 0.0398). However, mean allantoic fluid depth did not differ (P = 0.6120). There was a significant association between the body weight of calf and the diameter of the fetal aorta (P = 0.0115; r² = 0.3762). Surviving calves were lighter at birth (P = 0.0028) and were born later (P = 0.007) than NS calves. Calves born vaginally had a smaller fetal aorta (2.1 ± 0.1 cm vaginal and 2.4 ± 0.1 cm Cesarean) (P = 0.0487) and a lighter birth weight (41.4 ± 4.2 kg vaginal and 60.4 ± 2.1 kg Cesarean) (P = 0.0001) than calves born by Cesarean. Also, calves that underwent spontaneous labor (52.2% S and 0% NS; P = 0.0029) had a lighter birth weight (44.9 ± 3.8 kg) than calves that did not initiate labor (61.6 ± 2.2 kg) (P = 0.0004). Frequent ultrasonographic fetal monitoring allowed identification of differences between S and NS calves. Calves without a final decrease in FHR or with a large aortic diameter were more likely to require a Cesarean because of failure to initiate labor or fetomaternal disproportion. Parameters of fetal well-being and development during the last 3 weeks of gestation were first described in SCNT-derived beef calves.     

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.     

A retrospective analysis of factors contributing to calf mortality and dystocia in beef cattle

Records of 2191 calvings from the Clemson University Beef Physiology Unit between 1981 and 1993 were analyzed to determine factors affecting malpresentation, mortality and dystocia. Only 20 (0.91%) parturitions involved malpresentation: posterior presentation (n = 14), leg deviations (n = 3), head deviations (n = 2) and breech birth (n = 1). Dystocia affected calf mortality within 24 h of birth (P < 0.05), with mortality increasing as the severity of dystocia increased. There was an overall 4.5% death loss within 24 h of birth, with 4 and 7% mortality rates for calves from multiparous and primiparous dams, respectively (P < 0.05). Mortality was higher for bull vs heifer calves (P < 0.05). Ninety-four percent of calvings were unassisted, while 6% were assisted births. Dystocia was greater (P < 0.01) in primiparous (17%) than in multiparous dams (4%). In births involving dystocia, 28.1% required mild traction, 69.3% required heavy traction and 2.6% required Cesarean section. Birth weights associated with normal births and mild traction (36 and 36 kg) were lighter than those associated with heavy traction and Cesarean section (40 and 42 kg, respectively; P < 0.05). In conclusion, malpresentations were too few to be of significance, and dystocia influenced mortality within 24 h of birth. Calf birth weight and parity of dam explained most of the observed variations in dystocia.     

Lipid content and apoptosis of in vitro-produced bovine embryos as determinants of susceptibility to vitrification

The objective was to evaluate supplementation of fetal calf serum (FCS) and phenazine ethosulfate (PES), a metabolic regulator that inhibits fatty acid synthesis, in culture media during in vitro production (IVP) of bovine embryos. Taking oocyte fertilization (n = 4,320) as Day 0, four concentrations of FCS (0, 2.5, 5, and 10%) and three periods of exposure to PES (without addition—Control; after 60 h—PES Day 2.5 of embryo culture; and after 96 h—PES Day 4) were evaluated. Increasing FCS concentration in the culture media enhanced lipid accumulation (P < 0.05), increased apoptosis in fresh (2.5%: 19.1 ± 1.8 vs 10%: 28.4 ± 2.3, P < 0.05; mean ± SEM) and vitrified (2.5%: 42.8 ± 2.7 vs 10%: 69.2 ± 3.4, P < 0.05) blastocysts, and reduced blastocoele re-expansion after vitrification (2.5%: 81.6 ± 2.5 vs 10%: 67.3 ± 3.5, P < 0.05). The addition of PES in culture media, either from Days 2.5 or 4, reduced lipid accumulation (P < 0.05) and increased blastocoele re-expansion after vitrification (Control: 72.0 ± 3.0 vs PES Day 2.5: 79.9 ± 2.8 or PES Day 4: 86.2 ± 2.4, P < 0.05). However, just the use of PES from D4 reduced apoptosis in vitrified blastocysts (Control: 52.0 ± 3.0 vs PES Day 4: 39.2 ± 2.4, P < 0.05). Independent of FCS withdrawal or PES addition to culture media, the in vivo control group had lesser lipid accumulation, a lower apoptosis rate, and greater cryotolerance (P < 0.05). The increased lipid content was moderately correlated with apoptosis in vitrified blastocysts (r = 0.64, P = 0.01). In contrast, the increased apoptosis in fresh blastocysts was strongly correlated with apoptosis in vitrified blastocysts (r = 0.94, P < 0.0001). Therefore, using only 2.5% FCS and the addition of PES from Day 4, increased the survival of IVP embryos after vitrification. Moreover, embryo quality, represented by the fresh apoptosis rate, was better than lipid content for predicting embryo survival after vitrification.     

Variability of uterine blood flow in lactating cows during the second half of gestation

The main goal of the present study was to measure uterine blood flow volume (BFV) in the second half of gestation in lactating German Holstein cows. Furthermore, it was investigated, if there are individual variations in uterine blood flow and correlations between uterine blood flow and maternal weight and the birth weight of the calf. Forty-four cows were examined via color Doppler sonography in gestation weeks (GW) 21, 25, 29, 33, 37 and 39. The cows were allocated in groups based on the following variables: body weight (light ≤ 575 kg, heavy > 575 kg) and birth weight of the calf (light ≤ 42 kg, heavy > 42 kg). The BFV was measured via transrectal Doppler sonography of both uterine arteries. There was a linear increase in uterine BFV throughout the study period from 3053 ± 1143 ml/min to 16912 ± 5793 ml/min. Variation coefficients for inter-individual variations ranged from 34 to 37%. There was a moderate correlation between uterine BFV and birth weight of the calf in weeks 21 to 37 (0.30 ≤ r < 0.49; P < 0.05) and a good correlation in week 39 (r = 0.60; P < 0.0001). Uterine BFV in week 21 was significantly (P < 0.01) higher in heavy cows (3394 ± 1119 ml/min) than in light cows (2658 ± 1064 ml/min). Compared with light cows, the increase in uterine BFV was 32% higher in heavy cows during the study period. In week 21, there was no difference (P > 0.05) in uterine BFV between cows carrying a heavy calf (3351 ± 1130 ml/min) and those carrying a light calf (2796 ± 1115 ml/min). Thereafter, the increase of BFV was 43% higher in cows with a heavy calf than in those carrying a light calf. Cows with different body weights, but same birth weight of calf showed no differences (P > 0.05) in the increase of BFV, while in cows with the same body weight the rise in BFV was higher (P < 0.05) in those cows producing a heavy calf compared to cows carrying calves with light birth weights. In conclusion, there was a linear increase in uterine BFV in lactating Holstein cows during the second half of pregnancy with marked individual variations. Differences in the rise of BFV were more caused by the fetus than by body weight of cows.     

The efficiency of Afrikaner,Bonsmara, Simmentaler and hereford cattle in a sub-tropical environment

Norms for the performance of 4 breeds of cattle were studied. Bonsmara cows (434.8±3.5 kg) were the heaviest, followed by Simmentaler (418.7±3.9 kg), Afrikaner (411.6±4.0 kg) and Herefords (385.6±4.5 kg). Breeds differed significantly in terms of the preweaning growth of their calves. Simmentaler (212.3±2.6 kg) and Bonsmara (216.2±2.5 kg) were superior. A breed x age of cow interaction in terms of preweaning growth of the calf was established. Actual weaning mass of the calves appeared to have the most important influence on cow efficiency apart from weaning percentage.     

Calving difficulty and calf survival in beef cows fed two energy levels

Angus and Hereford yearling heifers (469) were bred to either Hereford or Angus bulls and fed either a high level (13.3 Mcal of ME) or a low level of nutrition (7.3 Mcal of ME) for the last 116 days of pregnancy. The level of calving difficulty did not differ between heifers receiving the high (H) and low (L) level of nutrition the first year (36% and 34%) but was significantly higher (P < .01) in heifers receiving H the second year (33% vs. 16%). Calves born to heifers receiving H weighed at birth 2.7 kg more (P < .01) both years than calves born to heifers receiving L. In the first year of the study the size of the pelvic opening was 10 cm larger 1 week before calving (P < .01) in heifers receiving H than in heifers receiving L. Calf survival was not related to level of nutrition, breed of dam or breed of sire either year of the study.     

Effects of severity of dystocia on cold tolerance and serum concentrations of glucose and cortisol in neonatal beef calves

Effects of dystocia on rectal temperature and serum cortisol and glucose concentrations, were studied in neonatal calves exposed to 0 degree C. Primiparous dams were observed continuously during parturition and if Stage II (labor) was not completed within 2 h after appearance of the allantochorion, delivery was completed with obstetrical assistance. Parturitions were scored (CDS) for difficulty and obstetric assistance required: CDS 1, no assistance (n = 8); CDS 2, minor manual assistance (n = 7); CDS 3, use of a mechanical calf puller (n = 5); CDS 4, cesarean section (n = 6). A blood sample, rectal temperature, and body weight were obtained within 30 min after birth. Calves were then fed 38 degrees C pooled colostrum, muzzled to prevent suckling, and placed back with their dam in a heated (22 degrees C) barn. At 4 h of age an indwelling jugular catheter was inserted. At 5 h of age calves were placed in a 0 degree C room for 140 min and blood samples and rectal temperatures were obtained every 10 or 20 min. A shivering score (1 = no shivering; 2 = moderate shivering; 3 = intense shivering) was assigned at each sampling time. Rectal temperatures were higher (P < 0.01) in CDS 1, 2 and 4 calves (39.0, 39.3, and 39.0 +/- .02 degrees C, respectively) than in calves with CDS 3 (38.3 +/- 0.02 degrees C) and were affected by duration of cold exposure (time; P < 0.01). Shivering was not affected by CDS but was affected by time (P < 0.01). Glucose concentrations were higher (P < 0.01) in CDS 3 calves (110.1 +/- 1.6 mg/dL) than in CDS 1, 2, or 4 calves (77.2, 86.4, and 89.0 +/- 1.3 mg/dL, respectively) and changed over time (P < 0.01). Cortisol concentrations were higher in CDS 1 calves (80.0 +/- 1.7 ng/mL) than in CDS 2, 3 or 4 calves (62.7, 58.2, and 57.7 +/- 2.0 ng/mL, respectively) and were affected by time (P < 0.01). We conclude that severe dystocia (CDS 3) resulted in lower calf rectal temperature, reduced serum cortisol, and increased serum glucose which could affect the ability of the calf to withstand cold stress. Minor dystocia did not cause and timely cesarean delivery prevented, the physiological aberrations encountered in severe dystocia.     

Factors associated with passive immunity transfer in dairy calves: combined effect of delivery time,amount and quality of the first colostrum meal

Despite the well-known importance of an adequate colostral immunoglobulin (Ig) transfer to calf health and survival, failed transfer of passive immunity (FTPI) remains a widespread problem in dairy farming. The aim of this study was to investigate the management factors associated with FTPI in newborn calves, evaluating particularly the combined effect of delivery time, amount and quality of the first colostrum meal. The study was conducted from March to August 2014 on 21 Italian dairy farms. Farmers were asked as first to answer a farm-level questionnaire on calf management. Blood sampling was then performed on overall 244 calves (1 to 5 days of age) born from Holstein cows, and a sample of the first colostrum meal of each calf was collected. Individual information on calves and the respective colostrum management were recorded. Serum and colostrum Ig concentrations were assessed by electrophoresis. A mixed effects multivariable logistic regression model was used to investigate the association of the variables obtained from both the management questionnaire and the individual calf data with FTPI (calf serum Ig concentration <10.0 g/l). A cumulative colostrum management score (CMS) that considered delivery time, amount and quality of the first colostrum meal was generated for 236 calves, with higher values indicating better colostrum management. Overall, 41.0% of the calves were found having FTPI, and within-farm percentage of FTPI was over 20.0% in 71.4% of the farms. The risk of having FTPI was higher both for Holstein purebred calves compared with Holstein-beef crossbreds and for females compared with males. Moreover, it increased by 13% with every hour of delay of the first colostrum meal provision since birth, whereas it decreased by 59% and 3%, respectively, with every additional liter of colostrum given and every additional gram of Ig per liter contained in the colostrum fed. Calf serum Ig concentration varied significantly according to the CMS, increasing by 1.53 g/l with every additional CMS point. In order to completely avoid FTPI, calves should receive at least 2.5 l of high-quality colostrum (Ig concentration >87.6 g/l) within 1.0 h from birth. Considerable improvements are still needed about colostrum management for newborn calves in dairy farms. The results of this study will help in developing farm-specific programs for reducing the occurrence of FTPI.     

Respiratory and cardiovascular effects of doxapram and theophylline for the treatment of asphyxia in neonatal calves

Respiratory stimulants are widely used in asphyxic neonatal calves despite a lack of data about their effectiveness and indications of possible side effects. The effect of doxapram and theophylline on respiratory, cardiovascular, and acid-base variables was investigated in 10 healthy neonatal calves (Bos Taurus). A venous, a peripheral arterial, and a pulmonary arterial catheter were placed, and central venous, pulmonary, and systemic blood pressures and cardiac output were measured using thermodilution technique. Doxapram, but not theophylline, led to an immediate increase in respiratory rate (P ≤ 0.01). The arterial pCO₂ decreased to 27.1 ± 4.7 mm Hg within 30 sec after doxapram administration and to 46.3 ± 5.8 mm Hg within 120 min after theophylline administration (P < 0.0001). The systolic pulmonary pressure increased from 70 ± 8 mm Hg (mean ± SD) to 93 ± 19 mm Hg within 30 sec after doxapram, but decreased after theophylline. The pulmonary vascular resistance also increased after doxapram and decreased after theophylline (P < 0.01). Doxapram had a more pronounced and faster effect on respiratory rate and elimination of CO₂ than theophylline. Doxapram, but not theophylline, is indicated for treatment of postnatal asphyxia in calves, but there are potential cardiovascular side effects.     

Traction force microscopy in rapidly moving cells reveals separate roles for ROCK and MLCK in the mechanics of retraction

Retraction is a major rate-limiting step in cell motility, particularly in slow moving cell types that form large stable adhesions. Myosin II dependent contractile forces are thought to facilitate detachment by physically pulling up the rear edge. However, retraction can occur in the absence of myosin II activity in cell types that form small labile adhesions. To investigate the role of contractile force generation in retraction, we performed traction force microscopy during the movement of fish epithelial keratocytes. By correlating changes in local traction stress at the rear with the area retracted, we identified four distinct modes of retraction. “Recoil” retractions are preceded by a rise in local traction stress, while rear edge is temporarily stuck, followed by a sharp drop in traction stress upon detachment. This retraction type was most common in cells generating high average traction stress. In “pull” type retractions local traction stress and area retracted increase concomitantly. This was the predominant type of retraction in keratocytes and was observed mostly in cells generating low average traction stress. “Continuous” type retractions occur without any detectable change in traction stress, and are seen in cells generating low average traction stress. In contrast, to many other cell types, “release” type retractions occur in keratocytes following a decrease in local traction stress. Our identification of distinct modes of retraction suggests that contractile forces may play different roles in detachment that are related to rear adhesion strength. To determine how the regulation of contractility via MLCK or Rho kinase contributes to the mechanics of detachment, inhibitors were used to block or augment these pathways. Modulation of MLCK activity led to the most rapid change in local traction stress suggesting its importance in regulating attachment strength. Surprisingly, Rho kinase was not required for detachment, but was essential for localizing retraction to the rear. We suggest that in keratocytes MLCK and Rho kinase play distinct, complementary roles in the respective temporal and spatial control of rear detachment that is essential for maintaining rapid motility.     

Feeding behaviour and activity as early indicators of disease in pre-weaned dairy calves

Across the industry, there is large variation in health status of dairy calves and as a result, disease incidence and antibiotic use is high. This has significant implications for animal welfare, productivity and profitability of dairy and dairy-beef production systems. Technology-based early detection systems could alleviate these issues; however, methods of early detection of disease in dairy calves have not been widely explored. This study aimed to determine whether changes in activity and feeding behaviour can be used as early warning indicators of respiratory disease in calves. In total, 100 pre-weaned male Holstein calves (age: ~ 8−42 days) were used. Calves were group-housed and provided with starter diet, straw bedding and ad libitum water. Calves were fed milk replacer ad libitum through an automatic calf feeder, and each calf was fitted with a leg-mounted activity monitor. Daily activity and feeding behaviour variables were calculated for each calf. Each calf was assessed daily using a modified version of the Wisconsin Scoring System to assess respiratory disease status. Calves were classed as ‘Diseased’, ‘Intermediate’ or ‘Healthy’ based on their cumulative health score. The peak day of the most extreme illness event was identified for each calf. Data from Diseased and Healthy calves were paired for analysis based on age and BW. Data were compared for the day of peak illness, and for the 3 days previous and post. Compared to healthy calves, diseased calves lay for longer and tended to have longer lying bouts (daily lying: 17.6 ± 0.3 vs 16.7 ± 0.2 h, P < 0.01; bout length: 74.8 ± 10.6 vs 56.0 ± 3.7 min, P = 0.09 for diseased and healthy calves, respectively). Diseased calves fed for a shorter time and had fewer feeder visits (with intake) each day compared to healthy calves (feeding time (min): 19.3 ± 1.4 vs 22.8 ± 1.5; P < 0.05; visits: 2.1 ± 0.2 vs 3.2 ± 0.4; P < 0.05). Importantly, differences between diseased and healthy calves were evident in both activity and feeding behaviour on the days prior to the peak day of disease. Lying bout length was greater in diseased calves for the 2 days prior to the peak day (P < 0.05), lying time was longer on day − 1 (P < 0.05) and feeder visits with milk intake were less frequent on day − 3 (P < 0.05). Thus, measurement of feeding and activity using precision technology within early detection systems could facilitate early intervention and optimized treatment.     

Characterization and cytogenetic analysis of Japanese Black calves produced by nuclear transfer

To confirm the normality of the Japanese Black calves produced by nuclear transfer, we examined the properties of such calves at parturition and analyzed their karyotypes. Thirty Japanese Black calves were produced by nuclear transfer; 3 of these calves (10.0%) required assisted delivery while 1 calf (3.3%) died soon after birth. Average birth weight was 31.0 +/- 1.8 kg and gestation period was 286.4 +/- 1.0 d (mean +/- SEM). None of the nuclear transfer calves showed external malformations. Within sets of cloned nuclear transfer calves, which were genetically identical, birth weights varied by up to 20.5 kg. Among singleton Japanese Black calves, the mean birth weight of nuclear transfer calves was significantly greater (P < 0.05) than that of calves produced by in vivo-derived embryo transfer. Cytogenetic analysis of 23 Japanese Black nuclear transfer calves revealed the presence of 2N 4N mosaicism in 21 of the nuclear transfer calves. The frequency of occurrence of tetraploidy was unrelated to birth weight. Endoreduplication was observed in 1 Japanese Black nuclear transfer calf, and the frequency of occurrence of the endoreduplication in this calf was 0.5% (1/209). We conclude that there was no external malformation or chromosomal aneuploidy in Japanese Black nuclear transfer calves, but the mean birth weight of nuclear transfer calves was heavier than that of in vivo-derived embryo transfer calves in both sexes, and a variation of birth weight within sets of nuclear transfer calves cloned from the same embryo was recognized.     

Influence of Predator Harvest,Biological Factors,and Landscape on Elk Calf Survival in Idaho

ABSTRACT We evaluated survival of elk (Cervus elaphus) calves on 2 contrasting study areas in north-central Idaho, USA, from 1997 to 2004. Recruitment was modest (>30 calves:100 F [calves of either sex: F elk 1 yr old]) and stable on the South Fork study area and low (<20 calves:100 F) and declining on the Lochsa study area. The primary proximate cause of calf mortality on both study areas was predation by black bears (Ursus americanus) and mountain lions (Puma concolor). We experimentally manipulated populations of black bears and mountain lions on a portion of each study area. Black bear harvest (harvest density/600km²) initially doubled on the Lochsa treatment after manipulating season bag limits. Mountain lion harvest also increased by 60% but varied widely during the manipulation period. Harvest seasons were closed for black bears and mountain lions on the treatment portion of the South Fork study area. Using the Andersen—Gill formulation (A-G) of the Cox proportional hazards model, we examined effects of landscape structure, predator harvest levels, and biological factors on summer calf survival. We used Akaike's Information Criterion (AIC_c) and multimodel inference to assess some potentially useful predictive factors relative to calf survival. We generated risk ratios for both the best models and for model-averaged coefficients. Our models predicted that calf survival was influenced by biological factors, landscape surrounding calf locations, and predator harvest levels. The model that best explained mortality risk to calves on the Lochsa included black bear harvest (harvest density/600 km²), estimated birth mass of calves, and percentage of shrub cover surrounding calf locations. Incorporating a shrub X time interaction allowed us to correct for nonproportionality and detect that effect of shrub cover was only influential during the first 14 days of a calf's life. Model-averaging indicated that estimated birth mass of calves and black bear harvest were twice as important as the next variables, but age of calves at capture was also influential in calf survival. The model that best explained mortality risk to calves on the South Fork included black bear harvest, age of calves at capture, and gender of calves. Model-averaging indicated that age at capture and black bear harvest were twice as important as the next variable, forest with 33–66% canopy cover (Canopy 33–66). Risk to calves decreased when calves occupied areas with more of this forest cover type. Model-averaging also indicated that increased mountain lion harvest lowered calf mortality risk 4% for every 1-unit increase in lion harvest (harvest density/600 km²) but was lower (<25%) in importance compared to age at capture and black bear harvest. Our results suggest that levels of predator harvest, and presumably predator density, resource limitations expressed through calf birth mass, and habitat structure had substantial effects on calf survival. Our results can be generalized to other areas where managers are dealing with low calf elk recruitment. However, because factors vary spatially, a single management strategy applied in different areas will probably not have the same effect on calf survival.     

Seasonal variations in the energy budget of Elliot’s pheasant (Syrmaticus ellioti) in cage

This study aimed to discuss the energy budget of Elliot’s pheasant Syrmaticus ellioti in different seasons, with life and health, good growth and normal digestion of Elliot’s pheasant as the tested objects, The energy budget of Elliot’s pheasant was measured by daily collection of the trial pheasants’ excrement in the biological garden of Guangxi Normal University from March 2011 to February 2012. The results showed that the gross energy consumption, metabolic energy and excrement energy varied by season, increasing as temperature decreased. There was significant difference in gross energy consumption, metabolic energy, excrement energy between adults and nonages. There was also a trend that food digestibility of pheasants increases as temperature increases. In the same season, the food digestibility of adults was better than that of nonages. Throughout spring, summer, autumn and winter, the metabolic energy of 4-year adults were 305.77±13.40 kJ/d, 263.67±11.89 kJ/d, 357.23±25.49 kJ/d and 403.12±24.91 kJ/d, respectively, and the nonages were 284.86±17.22 kJ/d, 284. 66±15.16 kJ/d, 402. 26±31.46 kJ/d and 420. 30±31.98 kJ/d, respectively. The minimum metabolic energies were 247.65±21.81 g, 265.86±26.53 g, respectively for each group, detected between 4-year adults and 1-year nonages. Further study is needed to determine whether 29.6 C is the optimal temperature for the Elliot’s pheasant.     

Deep intra-uterine artificial inseminations using cryopreserved spermatozoa in beluga (Delphinapterus leucas)

Artificial insemination (AI) with liquid-stored spermatozoa and sperm cryopreservation using directional freezing (DF) have been successful in the beluga. This study built on this foundation to develop a deep intra-uterine AI technique with frozen-thawed semen in beluga. Forty-two ejaculates from one male were cryopreserved using DF technology and subsequently used for 10 insemination attempts with seven females. Percentage pre- and post-thaw progressive motility and viability were (mean ± SD) 73.0 ± 12.2, 38.4 ± 8.8, 88.0 ± 0.1, and 59.3 ± 15.7%, respectively. A series of GnRH injections (3 x 250 μg, IV, 1.5 to 2 h apart) were used to induce ovulation, once a growing follicle >2.5 cm in diameter was visualized via trans-abdominal ultrasonography. Artificial insemination was performed at 30.1 ± 3.8 h post-initial GnRH injection with semen deposited in the uterine horn, 92.6 ± 16.2 cm beyond the genital opening using a flexible endoscope. The external cervical os (cEOS) was located beyond a series of 5 to 10 vaginal rings, 44.8 ± 9.3 cm from the external genital opening. The internal bifurcation of the uterus was 27 ± 6.8 cm beyond the cEOS. Ovulation occurred at 8.5 ± 7.6 h post-AI. Two of 10 inseminations (20%) resulted in pregnancy. The first pregnancy resulted in twins; both calves were born 442 d after AI, with one surviving. The second pregnancy is ongoing. These findings represent the first successful application of AI using frozen-thawed semen in beluga, and are important examples of how assisted reproductive technologies can provide tools for the global management of threatened species.     

Determining substrate displacement and cell traction fields--a new approach

This paper presents a new approach for the traction force microscopy (TFM) method which determines traction forces exerted by adherent cells on a thin, elastic polyacrylamide gel embedded with fluorescent microbeads. In this enhanced TFM method, a pattern recognition technique is first applied to match the pair of microbead embedded images before and after deformation, which subsequently provides the displacement field of the elastic substrate. Once the displacement field is obtained, the 3-D finite element method (FEM) is used to compute cell traction forces. The new TFM has been applied to determine traction forces of human tendon fibroblasts. Compared to existing TFM methods, the present method has the following advantages: (1) its displacement field obtained is associated with microbead movements; (2) it considers the finite thickness of the thin polyacrylamide gel and is therefore free from the infinite half-space approximation adopted by existing TFM methods; and (3) its computation procedure for determining cell traction forces is fast.