A possible role of central serotonin in L‐tryptophan‐induced GH secretion in cattle

To clarify the role of serotonin (5‐HT) in the regulatory mechanism of L‐tryptophan (TRP)‐induced growth hormone (GH) secretion in cattle, changes in 5‐HT concentrations in the cerebrospinal fluid (CSF) in the third ventricle (3V) and GH in plasma before and after the peripheral infusion of TRP were determined simultaneously. The direct effect of TRP on GH release from the dispersed anterior pituitary cells was also assessed. A chronic cannula was placed in 3V by stereotaxic surgery, then CSF and blood were withdrawn under physiological conditions. TRP (38.5 mg/kg BW) was infused through an intravenous catheter from 12.00 to 14.00 hours and CSF and blood sampling were performed from 11.00 to 18.00 hours at 1‐h intervals. The concentration of 5‐HT in CSF was determined by high‐performance liquid chromatography with electrochemical detection. GH, melatonin (MEL), and cortisol (CORT) concentrations were measured by radio‐immunoassay and enzyme‐immunoassay. Concentrations of 5‐HT were increased by TRP infusion. The TRP infusion significantly increased GH release. On the other hand, TRP did not stimulate GH release from the bovine pituitary cells. MEL and CORT concentrations were not altered by TRP infusion. These results suggest that TRP induced GH release via the activation of serotonergic neurons in cattle.     

Effect of animal age,postmortem chilling rate,and aging time on meat quality attributes of water buffalo and humped cattle bulls

The study was aimed to investigate the influence of animal age, post‐slaughter chilling rate, and aging time on meat quality of M. longissimus dorsi (LD) of water buffalo (Bubalus bubalis) and humped cattle (Bos taurus indicus) bulls. After slaughtering, one side of carcasses was subjected to rapid chilling (RC) (0 ± 2°C) and other side was hanged in controlled room temperature (25 ± 2°C) for 3 hr, then allowed to the chiller (0 ± 2°C). The meat quality traits were analyzed at 1, 7, and 14 days of storage. It was noted that rapidly chilled carcasses from the younger animals of both species missed the ideal pH/temperature window, which affects the toughness of the meat. Buffalo meat presented higher shear force, color L* values, and lower b* value as compared to the cattle meat. Moreover, meat shear force values decreased while all color coordinates and cooking loss values increased with lengthening the storage time in both age groups of cattle and buffalo. In conclusion, the tenderness of cattle meat was superior to that of buffalo and RC adversely affect the shear force values of young cattle and both age groups of buffalo bulls.     

Intravenous tryptophan administration attenuates cortisol secretion induced by intracerebroventricular injection of noradrenaline

This study was conducted to investigate the possibility of suppression of stress‐induced cortisol (CORT) secretion by tryptophan (TRP) administration and to better understand its regulatory mechanisms by using a noradrenaline (NA) injection into the third ventricle (3V) as a stress model in cattle. A total of 25 Holstein steers with a cannula in the 3V were used. First, the increase in CORT secretion was observed following a NA injection into the 3V in a dose‐dependent manner, verifying the appropriateness of this treatment as a stress model of CORT secretion (Experiment 1). The effect of prior‐administration of TRP into peripheral blood with a dose that has been demonstrated to increase brain 5‐hydroxytryptamine levels on the elevation of plasma CORT induced by NA or corticotropin‐releasing hormone (CRH) was then examined (Experiment 2). The prior administration of TRP suppressed NA‐induced, but not CRH‐induced, CORT elevation. These results suggest that an increase in TRP absorption into peripheral blood could suppress the stress‐induced CORT secretion in cattle via the attenuation of the stimulatory effect of NA on the hypothalamic CRH release.     

Comparison between the effects of feeding copper sulphate-treated and untreated rapeseed cake containing high glucosinolates on rumen fermentation,nutrient digestion and nitrogen metabolism in steers

Two trials were carried out to study the effects of copper sulphate (CuSO₄) on detoxifying glucosinolates (GLS) in rapeseed cake (RSC) and compare the effects of feeding CuSO₄-treated and untreated RSC on nutrient digestion and nitrogen (N) metabolism in steers. In Trial 1, different concentrations of CuSO₄ solution (1.6 vs. 3.2 g CuSO₄·5H₂O L⁻¹), soaking temperatures (25 vs. 60°C) and drying methods (air drying at 60°C vs. freeze drying) were allocated in a 2 × 2 × 2 factorial arrangement in vitro. In Trial 2, six steers and dietary inclusions of untreated RSC (control), CuSO₄-treated RSC and CuSO₄-added RSC were assigned in a replicated 3 × 3 Latin square design. CuSO₄ treatment in vitro decreased the contents of GLS and thiocyanate (TC) in RSC (p < 0.001). The total amount of GLS and TC decreased by 62.7–68.5% for all treatments. The animal trial showed that CuSO₄-treated RSC inclusion decreased ruminal concentration of valerate (p < 0.01), whereas it did not affect ruminal pH, ammonia N and total volatile fatty acids. Compared with the control, feeding CuSO₄-treated or CuSO₄-added RSC had no effect on plasma concentrations of triiodothyronine and thyroxine, N excretion and N retention. CuSO₄-treated RSC tended to increase neutral detergent fibre digestibility (p = 0.072) and urinary excretion of urea (p = 0.056). Urinary excretion of purine derivatives (p = 0.076) and rumen microbial N supply (p = 0.084) tended to decrease when feeding CuSO₄-treated RSC versus control. TC was found to be the only metabolite of GLS in rumen fluid, plasma and urine. It was feasible to detoxify GLS in RSC using low CuSO₄ at room temperature. However, feeding CuSO₄-treated or CuSO₄-added RSC had minor effects on rumen fermentation, nutrient digestion and N metabolism in steers. CuSO₄ treatment on RSC for feeding steers seems to be unnecessary.     

Effect of different penetrating and non‐penetrating cryoprotectants and media temperature on the cryosurvival of vitrified in vitro produced porcine blastocysts

The aim of this study was to determine the most efficient vitrification protocol for the cryopreservation of day 7 in vitro produced (IVP) porcine blastocysts. The post‐warm survival rate of blastocysts vitrified in control (17% dimethyl sulfoxide + 17% ethylene glycol [EG] + 0.4 mol/L sucrose) and commercial media did not differ, nor did the post‐warm survival rate of blastocysts vitrified in medium containing 1,2‐propandiol in place of EG. However, vitrifying embryos in EG alone decreased the cryosurvival rate (55.6% and 33.6%, respectively, p < .05). Furthermore, the post‐warm survival rates of blastocysts vitrified with either trehalose or sucrose as the non‐penetrating cryoprotectant did not differ. There was also no significant difference in post‐warm survival of blastocysts vitrified in control (38°C) media and room temperature (22°C) media with extended equilibration times, although when blastocysts were vitrified using control media at room temperature, the post‐warm survival rate increased (56.8%, 57.3%, 72.5%, respectively, p < .05). The findings show that most cryoprotectant combinations examined proved equally effective at supporting the post‐warm survival of IVP porcine blastocysts. The improved post‐warm survival rate of blastocysts vitrified using media held at room temperature suggests that the cryoprotectant toxicity exerted in 22°C media was reduced.     

Cold agglutinin activity in 2 dogs

A 5‐year‐old neutered male Mastiff and an 8‐year‐old spayed female Labrador Retriever were presented to the University of Minnesota Veterinary Medical Center. The Mastiff was presented for evaluation of lameness and pyoderma one month prior in Missouri, where he tested positive for Ehrlichia canis by serum ELISA test, treated with doxycycline. PCR for Ehrlichia sp, Anaplasma sp, Babesia sp, and Bartonella sp, and PCR for antigen receptor rearrangement were negative, serum protein electrophoresis (SPE) revealed polyclonal gammopathy, and mildly reactive lymphoid cells were seen cytologically. The Labrador presented with a proliferative rostral mandibular gingival mass and lipomas for further presurgical evaluation of cold agglutinin activity documented by a commercial laboratory 2 years earlier prior to removal of a grade II mast cell tumor. This dog had a negative SNAP4Dx, normal SPE, and persistently increased serum ALP activity and polyuria/polydipsia suggestive for hyperadrenocorticism. Both dogs had markedly agglutinated RBC in the EDTA samples that dispersed with warming, and normal plasma color. Cold agglutinin activity was demonstrated by direct saline agglutination testing using whole blood and washed erythrocytes demonstrating agglutination at 30°C, 25°C, 15°C, and 4°C, but not at 37°C . CBC results (ADVIA 2120i) from the Mastiff revealed no significant differences in the RBC results obtained at room temperature (RT) and at 37°C; however, the RT run demonstrated negative bias in neutrophil and platelet concentrations attributed to rapid RBC settling. This uncommon hematologic condition may cause artifacts on the automated leukogram and platelet count, and may be subclinical for long periods.     

Effect of health status on fattening performance in young crossbred polish Holstein‐Friesian × Limousin Bulls and steers

The aim of this study was to determine the effect of disease incidence on selected parameters of cattle fattening performance and carcass quality, and the fatty acid profile of beef. The experimental materials comprised 16 bulls and 16 steers, Polish Holstein‐Friesian × Limousin crossbreeds (including 10 healthy and six treated animals of each category). At 5 weeks of age, bloodless castration was carried out using a rubber elastrator. The calves were fed milk replacer provided in automatic feeding stations. Until 540 days of age, the animals were fattened in an Animal Research Laboratory equipped with the Roughage Intake Control (RIC) system (Insentec, the Netherlands). In comparison with healthy (untreated) bulls and steers, sick (treated) animals had lower average body weight at 180 days of age, by 37 kg (P ≤ 0.05) and lower average final body weight at 540 days of age, by 56 kg (P ≤ 0.05). Sick animals were characterized by lower feed intake and worse feed efficiency (not statistically significant differences). Hot carcass weight reached 318 kg in healthy animals and 258 kg in treated bulls (P ≤ 0.05). In treated steers, the percentage of lean meat and bones in the three‐rib section was higher and the percentage of fat was lower, compared with their healthy counterparts (P ≤ 0.01). There was a category × health status interaction for carcass tissue composition. There were no significant influences of type of sickness on analyzed traits. In comparison with healthy steers, intramuscular fat of Musculus longissimus dorsi (MLD) from treated steers had significantly (P ≤ 0.05) higher concentrations of polyunsaturated fatty acids (n‐6 and n‐3) and a lower content of conjugated linoleic acid.     

Successful Cryopreservation of Domestic Cat (Felis catus) Epididymal Sperm after Slow Equilibration to 15 or 10°C

To support conservation strategies in wild species, simple but highly reproducible procedures of sperm cryopreservation are required for an application under field conditions. We used epididymal sperm of the domestic cat to optimize a sperm freezing procedure for felid species, particularly questioning the demand for sperm cooling to 4°C. We equilibrated sperm during slow cooling to only 15 or 10°C in a Tes–Tris–fructose extender with final concentrations of 4.7% (v/v) glycerol and 10% (v/v) of the water‐soluble fraction of hen's egg yolk (low‐density lipoproteins). Subsequently, sperm were frozen over liquid nitrogen. Total and progressive motility (mean ± SD) after thawing was 60.7 ± 8.6% and 53.9 ± 9.6% in samples cooled to 15°C or 61.6 ± 9.5% and 55.3 ± 9.9% in samples cooled to 10°C. Therefore, a one‐step addition of glycerol to sperm at room temperature together with the freezing extender, the use of cryovials (loaded with diluted sperm aliquots of 300 μl), an equilibration period of 40 min comprising slow cooling to 15°C at a rate of approximately ?0.14 K/min before rapid freezing over liquid nitrogen, yielded satisfying results. Cooling, freezing and thawing rates were exactly characterized as a prerequisite for further optimization and to provide a repeatable protocol to other practitioners.     

Characterization of adipose-derived equine and canine mesenchymal stem cells after incubation in agarose-hydrogel

Adult stem cells are of particular interest for the therapeutic approach in the field of regenerative medicine. Due to their ease of harvest, adipose-derived mesenchymal stem cells (ASCs) are an attractive stem cell source that has become increasingly popular. Critical aspects of applied cell therapies are the circumstances of transport from the laboratory towards the site of operation and cell delivery into the desired area. With regard to these issues, agarose-hydrogel was analyzed as a cell carrier matrix of equine and canine ASCs in vitro, which can be used for minimally invasive application. Isolated ASCs were expanded and 2.5 × 10⁶ cells were combined with agarose-hydrogel to build a 0.4% hydrogel-cell solution which was stored at two temperatures (room temperature (RT) vs. 37°C). Cell viability was investigated (live-dead assay) at different time points (0, 1, 6 and 24 h) in order to determine i) the effect of different temperatures on the cell survival as well as ii) the maximum possible time span before implantation. CFU-assay and WST-1 assay were performed after 24 h incubation in agarose-hydrogel and the cells were induced into adipogenic and osteogenic differentiation to analyze the effects of the incubation on the cell behaviour. No negative effect of the agarose-hydrogel incubation was determined on the different species’ cell behaviour at either RT or 37°C with any of the assays used. We can recommend agarose-hydrogel as a cell carrier for cell implantation with a storage period of up to 24 h at room temperature or at 37°C prior to implantation.     

Effect of the Depth of Insertion of the Thermometer on the Rectal Temperature of Donkeys During the Hot-Dry Season in a Tropical Savannah

The objective of the present study was to evaluate the effect of the depth of insertion and environmental parameters on the rectal temperature (RT) in donkeys during the hot-dry season in a tropical savannah zone of Nigeria. The experimental subjects were comprised of thirty donkeys divided into three groups based on age: group I, 10 foals (40.67 ± 2.20 kg; 1.50 ± 0.02 months); group II, 10 yearlings (91.53 ± 0.54 kg; 1.51 ± 0.01 years); and group III, 10 adults (140 ± 0.71 kg; 8.03 ± 0.06 years). Each group was divided into 5 male and 5 female donkeys. Measurements of the RT were recorded with a digital thermometer probe (model HI935007, Hanna Instruments), which was inserted into the rectum at varying depths of 3.5, 7, 10.5, and 14 cm in the same animal in each group. There was a gradual increase in the RT in donkeys as the depth of insertion was increased from 3.5 cm (36.60°C) to 14 cm (38.40°C). Data obtained from the study were subjected to repeated-measures analysis of variance, followed by Tukey's post-hoc test to compare mean values between different depths of RT measurements. Overall, there was a variation in the RT by the depth of insertion with the shallow depth of 3.5 cm having a lower RT than the depths of 7, 10.5, and 14 cm. The variation of the RT observed in donkeys showed that there is need to standardize the probe-insertion depth in veterinary clinical practice for accurate measurement of the RT in donkeys in the Northern Guinea savannah zone of Nigeria.     

The use of gelatine in long‐term storage (up to 48 hr) at 5°C preserves the pre‐freezing and post‐thawing quality of brown bear sperm

Sedimentation of spermatozoa occurs during long‐term liquid storage and this may produce deleterious changes. Our aim was to apply gelatine supplementation during long‐term pre‐freezing storage of bear sperm, applying final dilution and 6% glycerol at room temperature and cool in straws. We tested four models of sperm storage using a 1:1 dilution in TTF‐ULE‐Bear extender (TesT‐fructose‐egg yolk‐glycerol 6%): (i) second 1:1 dilution at room temperature (RT), cooling at 5°C in a tube and final dilution (100 × 10⁶ sperm ml^?1) (Standard); (ii) final dilution at RT and cooling in a tube (FD‐Tube); (iii) final dilution at RT and cooling in 0.25 ml plastic straw (FD‐Straw); and (iv) final dilution at RT in extender supplemented with 1.5% gelatine (Gelatine) and cooling in a 0.25 ml plastic straw. A Standard sample was stored at 5°C for 1 hr (Control); the rest of the samples (Standard, FD‐Tube, FD‐Straw, Gelatine) were stored for 24 or 48 hrs before freezing (100 × 10⁶ sperm ml^?1, glycerol 6%). The quality of the samples was assessed for motility by CASA, and viability (SYBR‐14/propidium iodide‐PI‐; VIAB), acrosomal status (PNA‐FITC/PI; iACR) and apoptotic status (YO‐PRO‐1/PI; YOPRO‐) by flow cytometry. At pre‐freezing, after 48 hr, Gelatine showed significantly higher viability (for VIAB and YOPRO‐) and progressiveness (PM, LIN and STR). At 48 hr, Gelatine showed similar YOPRO‐, iACR, LIN, STR and ALH respect to Control. At both 24 and 48 h post‐thawing, Gelatine sample had similar scores for YOPRO‐, iACR, LIN, STR, WOB and VIAB (only 24 hr) when compared with Control, and lower for TM, PM, rapidPM, VAP and ALH. No differences were found among others experimental groups with respect to Control. In conclusion, gelatine could be a suitable alternative to preserve the viability and progressive motility of brown bear ejaculates during long‐term pre‐freezing storage at 5°C.     

Effects of Temperature on In Vitro Short‐Term Storage of Sterlet Sturgeon (Acipenser Ruthenus) Ova

Artificial propagation of sturgeons is becoming increasingly important for recovery efforts as well as for commercial production. Sterlet Acipenser ruthenus is a common Eurasian sturgeon with a small body size and one of the fastest reproductive cycles among the sturgeons. The practical question being addressed in this study was how long fertilization of ovulated eggs can be delayed without substantially reducing the hatching rate, and an ancillary question is under what' temperature conditions do eggs retain good quality. Broodstock were injected with homogenized carp pituitary extract (CPE); ovulated eggs from three females were allocated to various treatment groups for temperature storage (control, 7°C, 11°C, 15°C and 19°C) until fertilized. Storage times at the regulated temperatures prior to fertilization were for 2.5, 5.0, 7.5 and 10.0 h. After the selected storage times in ovarian fluid, eggs were fertilized and transferred to incubation cages and then they were counted. Three replicates were allocated to each storage period and temperature. Hatched larvae were counted at 7‐day post‐fertilization. We found that sterlet eggs do not need to be fertilized immediately after collection. Reasonably good quality was retained for several hours if temperature conditions are fairly cool and stable. Eggs retained good quality when stored at 7°C and 11°C for up to 10 h with 54.1 ± 2.9 to 69.9 ± 7.9% hatching success, but egg quality was significantly reduced after 5‐h storage at 19°C (p < 0.01) and 7.5‐h storage at 15°C (p < 0.05) compared to cooler temperatures. Uniform temperatures between 7°C and 11°C can be considered as appropriate for storage of eggs in ovarian fluid for up to 10 h. This information can have practical application to routine hatchery practice for acipenserids, as well as for certain research protocols.     

Seasonal and sex differences in area preference and behavior of young cattle just after long distance transport

To determine seasonal and sex differences in behavioral motivation of cattle just after long transport, 54 Japanese Black × Holstein cattle were observed at 5 min intervals for 2 h just after 25‐h transport by road and ferry. The stocking pen (12.0 × 9.5 m) consisted of an eating area (near a feeding alley), a drinking area (near water bowls) and a resting area (all areas except for other two areas). First, the effect of season was determined with heifers in summer (n = 12: 7.9 ± 0.6 months of age; 292.0 ± 18.5 kg) and autumn (n = 19: 8.2 ± 0.6 months of age; 295.8 ± 15.5 kg). The mean temperature on the observation day was 28.5 (max: 34.5, min: 24.5)°C in summer and 20.3 (max: 26.5, min: 16.4)°C in autumn. Percentage of cattle staying in each area was different by season (χ² = 22.0; P < 0.01). In summer, the percentage of cattle staying in the drinking area (26.7%) was greater than the expected percentage (16.7%). Percentage of cattle staying in the eating area was greater in both seasons (31.3% in summer and 53.6% in autumn) than the expected percentage (16.7%). However, the mean percentage of cattle performing each behavior was not significantly different by season. Secondly, the effect of sex was determined with steers (n = 23: 7.6 ± 0.6 months of age; 301.9 ± 20.7 kg) and heifers (n = 19: same as above) in autumn. Although the percentage of cattle staying in each area was different by sex (χ² = 20.2; P < 0.01), the percentages of steers (25.5%) and heifers (53.6%) staying in the eating area were both greater than the expected percentage (16.7%). However, percentage of animals performing each behavior was not different by sex. These results recommend to stockpersons that they should install additional troughs for hay and water into a pen just after long distance transport, since the number of cattle that can eat and drink at the same time was limited.     

Gender status effect on carcass and meat quality traits of feedlot Angus × Nellore cattle

The study evaluated the effect of gender status on carcass and meat quality of feedlot Angus × Nellore cattle. A total of 176 cattle, 20 months old, were confined for 190‐days and assigned to four treatments: bulls, immunocastrated, steers, and heifers. Bulls had greater rib eye area and HCW (p = 0.0001). Heifers had increased fat thickness (p = 0.0001). Steers and heifers had higher marbling scores (p = 0.0001). There was interaction between gender and aging time for Warner‐Bratzler Shear Force (p = 0.0002), L* (p = 0.0118), and b* (p = 0.0113) values of beef. The sensory panel results showed that beef from bulls had the lowest consumer overall acceptance (p = 0.0278). Especially, regardless tenderness, steers and immunocastrated beef were considered tender, independent of aging time. Beef produced by heifers, steers, and immunocastrated is considered to be of higher quality than bulls. Thus, it is may be an interesting alternative to produce high‐quality beef than bulls, to attend the consumer demand for high‐quality products. Additionally, the low fatty acids n6 levels and low n6:n3 ratio, high levels of CLA, MUFAs, and oleic acid suggests that the heifer meat is favorable for human health.     

Effect of olive-derived antioxidants (3,4-dihydroxyphenylethanol and 3,4 dihydroxyphenylglycol) on sperm motility and fertility in liquid ram sperm stored at 15°C or 5°C

The aim of the present study was to evaluate the effect of the addition of two olive oil-derived antioxidants, hydroxytyrosol (3,4-dihydroxyphenylethanol, HT) and 3,4-dihydroxyphenylglycol (DHPG), on ovine semen during liquid storage at 5°C and 15°C. Semen was collected, pooled, diluted and then divided into aliquots supplemented with different concentrations (5 μg/ml, 10 μg/ml, 50 μg/ml and 100 μg/ml) of HT, DHPG and a mixture (MIX) of both antioxidants. Sperm motility characteristics were assessed in the different samples at 0, 6, 24, 48, 72 and 96 hr after cooling, and a fertility trial was also conducted. The results showed that the antioxidant addition did not significantly improve total and progressive motility in ovine cooled sperm maintained at 15° or 5°C. However, in samples stored at 5°C, LIN (48, 72, 96 hr), STR (0 hr) and WOB (0, 48, 72, 96 hr) values significantly decreased in comparison with control treatment when high antioxidant concentrations were added (MIX100 or HT100). When samples were maintained at 15°C, MIX50 showed significantly higher VCL values than the control treatment after 6 hr cooling, and MIX100 showed significantly lower VCL values at 96 hr after cooling. According to the artificial insemination trial, no significant differences were observed when antioxidants were added. In conclusion, the use of HT and DHPG showed small impact in sperm motility and fertility was not affected (nor detrimentally nor positively) when insemination was carried out using antioxidant-supplemented liquid sperm.     

Effects of Retinoids on the In Vitro Development of Capra Hircus Embryos to Blastocysts in Two Different Culture Systems

The objective of this study was to evaluate the effect of retinol (RT) and retinoic acid (RA) on the in vitro development of pre‐implantation goat embryos cultured in potassium simplex optimized medium or synthetic oviduct fluid or cocultured in oviductal cells monolayer either in potassium simplex optimized medium or synthetic oviduct fluid. A total of 2407 cumulus‐oocyte complexes were aspirated from 2 to 6 mm ovarian follicles from slaughtered animals. Selected cumulus‐oocyte complexes were subjected to in vitro maturation in TCM 199 for 24 h at 39°C in an atmosphere of 5% (v/v) CO₂ in humidified air. In vitro fertilization was performed in modified defined medium. Eighteen hours after in vitro fertilization, cumulus cells were removed and presumptive zygotes were randomly distributed into experimental groups. In Experiment 1, presumptive zygotes were cultured in potassium simplex optimized medium, potassium simplex optimized medium + RT, potassium simplex optimized medium + retinoic acid, synthetic oviduct fluid, synthetic oviduct fluid + RT and synthetic oviduct fluid + RA at 39°C in a humidified atmosphere of 5% (v/v) CO₂, 5% (v/v) O₂ and 90% (v/v) N₂. In Experiment 2, presumptive zygotes were cocultured in potassium simplex optimized medium + oviductal cells monolayer, potassium simplex optimized medium + RT + oviductal cells monolayer, potassium simplex optimized medium + RA + oviductal cells monolayer, synthetic oviduct fluid + oviductal cells monolayer, synthetic oviduct fluid + RT + oviductal cells monolayer and synthetic oviduct fluid + RA + oviductal cells monolayer in an atmosphere of 5% (v/v) CO₂ in humidified air. In both experiments, media were partially changed on day 2 after in vitro fertilization and unfertilized oocytes were excluded from the experiment. Embryos were cultured or cocultured for 8 days. In Experiment 1, there was no effect of RT or RA supplementation on the proportion of oocytes that reached the morula or blastocyst stages. By contrast, Experiment 2 demonstrated that the addition of 0.28 μg/ml RT and 0.5 μm RA to the embryo culture media stimulated (p < 0.05) development to the morula and blastocyst stages under the coculture conditions tested. In conclusion, retinoids play an important role in pre‐implantation development of goat embryos and can be used to enhance in vitro embryo production.     

Effect of regular walking exercise on glucose tolerance and insulin response to i.v. glucose infusion in growing beef steers

The purpose of the present paper was to investigate the effect of regular walking exercise on glucose tolerance and insulin response to i.v. glucose infusion in growing beef steers. Four crossbred beef steers walked on a treadmill during a 6 week exercise period (1.2 km/h, 1 h/day and 5 days/week). The changes in plasma glucose and insulin levels following glucose infusion were analyzed immediately prior to (bodyweight: 260.4 ± 24.2 kg) and after (295.7 ± 30.1 kg) the exercise period. The basal levels of plasma glucose (86.4 vs. 82.0 mg/dL, P = 0.040) and insulin (24.5 vs. 14.3 μU/mL, P = 0.016) were significantly lower after the exercise period. Further, the increase in the levels of plasma glucose (420.4 vs. 280.8 mg/dL, P < 0.001) and insulin (94.5 vs. 73.1 μU/mL, P = 0.028) following the glucose infusion decreased after the exercise period. The area under the curve of plasma glucose (108.8 vs. 62.9 mg/dL per min, P < 0.001) and insulin (53.6 vs. 29.7 μU/mL per min, P = 0.018) indicated more rapid clearance of exogenous glucose and less insulin secretion for glucose clearance after the exercise period. These results suggest that regular exercise improves glucose tolerance, with lower insulin response to glucose infusion in growing steers, as observed in rodents and humans.     

Bovine acute-phase response after different doses of corticotropin-releasing hormone challenge

The objective was to compare the acute-phase response of steers receiving different doses of bovine corticotropin-releasing hormone (CRH). Fourteen weaned Angus steers (BW = 191 ± 2.1 kg, age = 167 ± 4.7 d) fitted with an indwelling jugular catheter and a rectal temperature (RT) monitoring device were assigned to receive 1 of 3 treatments (intravenous infusion): 1) 0.1 μg of CRH/kg of BW (CRH1; n = 5), 2) 0.5 μg of CRH/kg of BW (CRH5; n = 5), and 3) 10 mL of saline (0.9%; n = 4). Blood samples were collected via catheters, relative to treatment infusion (0 h), hourly from -2 to 0 h and 4 to 8 h and every 30 min from 0 to 4 h. Rectal temperatures were recorded every 30 min from -2 to 8 h. Blood samples were also collected via jugular venipuncture and rectal temperatures assessed using a digital thermometer every 6 h from 12 to 72 h and every 24 h from 96 to 168 h. All plasma samples collected during the study were analyzed for concentrations of haptoglobin. All plasma samples collected from -2 to 8 h were analyzed for cortisol concentrations. Serum samples collected hourly from -2 to 8 h were analyzed for concentrations of NEFA, IL-6, tumor necrosis factor (TNF)-α, and interferon-γ. Cortisol peaked at 0.5 h for CRH1 steers but returned to baseline concentrations at 1 h relative to infusion (time effect; P < 0.01). In CRH5 steers, cortisol peaked at 0.5 h and returned to baseline concentrations 3.5 h relative to infusion (time effect; P < 0.01). Cortisol concentrations did not change after treatment infusion for saline steers (time effect; P = 0.42). In CRH1 steers, NEFA concentrations peaked 5 h after treatment infusion (time effect; P = 0.01). Conversely, serum NEFA concentrations did not change for CRH5 and saline steers after treatment infusion (time effect; P > 0.37). Mean serum TNF-α concentrations in CRH1 steers after treatment infusion were greater compared with saline (P = 0.02), tended to be greater (P = 0.08) compared with CRH5, and were similar (P = 0.40) between CRH5 and saline steers. Mean RT in CRH1 steers after treatment infusion were greater (P < 0.04) compared with saline and CRH5 and similar (P = 0.50) between CRH5 and saline steers. Haptoglobin increased and peaked 72 h after treatment infusion for CRH1 steers (time effect; P = 0.01) but did not change for CRH5 and saline steers (time effect; P > 0.45). In conclusion, the bovine acute-phase response stimulated by CRH infusion is dependent on the CRH dose and the subsequent response in circulating cortisol.     

Effect of intracerebroventricular injections of prolactin‐releasing peptide on prolactin release and stress‐related responses in steers

Some evidence suggests that there might be a species difference in the effect of intracerebroventricularly administered (ICV) prolactin‐releasing peptide (PrRP) between rodents and sheep. We compared the levels of cortisol (CORT) and prolactin (PRL), rectal temperature (RT) and behavioral responses to ICV bovine PrRP (bPrRP) in steers. ICV bPrRP (0.2, 2 and 20 nmol/200 µL) tended to evoke a dose‐related increase in CORT concentrations and 0.2 nmol of bPrRP induced transient increase in PRL concentrations. A significant time–treatment interaction was observed for the percent change of CORT (P < 0.05) and PRL (P < 0.05) from pre‐injection value. The time–treatment interaction for changes in RT was not significant (P = 0.50). There tended to be a difference among the four treatments in terms of maximum change in RT from the pre‐injection value between 0 and 90 min (P < 0.1). Stress‐related behavioral signs were not observed in the present experiment. These findings indicate that ICV bPrRP increased CORT and PRL levels, suggesting that central PrRP might participate in controlling the hypothalamo‐pituitary‐adrenal axis and PRL release in cattle, unlike sheep. In contrast, central PrRP is unlikely to be involved in controlling the behavior of this species because ICV bPrRP did not induce marked changes in their behavior.     

Bovine growth hormone gene polymorphism affects stress response in Japanese Black cattle

We investigate the associations between growth hormone (GH) gene polymorphism and behavioral and physiological responses to stressors and learning ability in Japanese Black cattle. Flight distance test was conducted in the first experiment. Steers with haplotype C of GH gene polymorphism avoided human approaches at a significantly greater distance than ones without haplotype C (C: 1.9 ± 0.9, non‐C: 1.0 ± 0.2 m, P < 0.05). An open‐field test was conducted in the second experiment. Behavioral responses did not differ significantly between steers with and without haplotype C. Increases of heart rates to dropping of iron pipes was significantly higher in steers with haplotype C (C:161.7 ± 21.8, non‐C:130.7 ± 31.3%, P < 0.05). Despite basal serum concentrations not being different between steers with and without haplotype C, serum cortisol in blood sampling immediately after severe confinement in a race tended to be higher in steers with haplotype C (P = 0.1). The maze test was conducted as the third experiment. There was no difference in performance in the maze test between steers with and without haplotype C. It is concluded that genetic polymorphism of GH may affect stress responses through GH concentration in steers.