Short communication: Relationships among temperature-humidity index with rectal, udder surface, and vaginal temperatures in lactating dairy cows experiencing heat stress.

The objective of this study was to evaluate relationships between measurements of temperature-humidity index (THI) and rectal, vaginal, and udder surface temperatures in lactating cows exposed to heat stress (HS). In experiment 1, 12 multiparous and 8 primiparous Holstein cows experienced a THI ranging from 69 to 76 at 2000 to 1000 h and THI from 74 to 82 at 1000 to 2000 h (peaked at 82 from 1400 to 1800 h). Cows were exposed to HS 10 h daily for 21 d. Measurements of rectal temperature (RT) and udder surface temperature were collected at 1000 and 1500 h (±30 min). Vaginal temperature was monitored every 10 min using digital loggers, averaged over 1 h, and paired with corresponding rectal and udder surface temperature data. In experiment 2, 12 multiparous Holstein cows experienced a THI ranging from 60 to 76 at 2000 to 1000 h and THI from 69 to 83 at 1000 to 2000 h (peaked at 83 from 1600 and 1900 h), eliciting 10 h/d of HS for 7 d. Rectal and udder surface temperatures were analyzed at 0700 and 1500 h (±30 min). Vaginal temperature was recorded and analyzed as indicated in experiment 1. Afternoon THI showed weak correlations with surface temperature (r = 0.19, n = 420 in experiment 1; r = 0.23, n = 84 in experiment 2), weak to moderate correlations with RT (r = 0.34, n = 366 in experiment 1; r = 0.26, n = 84 in experiment 2), and moderate correlations with vaginal temperature (r = 0.34, n = 175 in experiment 1; r = 0.35, n = 40 in experiment 2). Moreover, vaginal temperature increased 0.10 and 0.22°C per unit of THI (R2 = 0.15 in experiment 1; R2 = 0.40 in experiment 2). Afternoon vaginal temperature strongly correlated with RT (r = 0.69, n = 131 in experiment 1; r = 0.63, n = 37 in experiment 2) and explained 57 (experiment 1) and 68% (experiment 2) of variation in RT. Surface temperature showed moderate to strong correlations with RT (r = 0.57, n = 84) and vaginal temperature (r = 0.74, n = 37) in experiment 2. In conclusion, THI showed a weak to moderate relationship with core body temperatures and explained the increase in rectal and vaginal temperatures experienced by HS cows. Compared with rectal temperature, vaginal temperature showed stronger relationships with THI and can be used to determine thermal load. Udder surface temperature showed a moderate to strong relationship with core body temperature, and this relationship may support the use of surface temperature data to manage thermal load in HS cows.

Bovine intramammary infection associated immunogenic surface proteins of Streptococcus uberis.

In spite of the increasing prevalence of Streptococcus uberis mastitis, its pathogenesis and associated virulence factors are not clearly defined. The aim of this study was to identify virulence associated genes and their products that can be used to develop effective vaccine to control bovine S. uberis mastitis. S. uberis was co-cultured with primary bovine mammary epithelial cells (PBMEC) or infused into mammary gland of dairy cows. The messenger RNA (mRNA) from S. uberis associated with PBMEC after 2 h or 4 h of co-culture was purified and sequenced. Results showed that virulence-associated genes such as surface lipoprotein (slp), infection induced histidine kinase (iihK), infection induced response regulator (iirR) and extracellular sugar binding protein 1 and 2 (exsbP1 and exsbP2) were among the top-up-regulated genes. To verify this observation in vivo, quantitative real time PCR (qRT - PCR) was conducted on mRNA of S. uberis recovered from milk of infected mammary glands 24 h post infection. Results revealed that in vitro up-regulated virulence-associated genes were also significantly up regulated under in vivo conditions. The iihK and iirR are flanked by exsbP1 and exsbP2 genes and this entire operon seems to be involved in adaptation to glands micro-environment, survival and colonization of the bovine mammary glands. Based on immunogenic epitope prediction of proteins encoded by these up-regulated genes during early stages of host-bacterial interactions slp, exsbP1 and exsbP2 genes were selected, cloned and expressed in E. coli. The purified recombinant proteins (rSlP, rExsbP1 & rExsbP2) reacted strongly with convalescent serum from cows experimentally infected with S. uberis confirming that they are immunogenic. These proteins may serve as potential targets to develop an effective vaccine against S. uberis mastitis.

Short communication: A reproductive tract scoring system to manage fertility in lactating dairy cows.

We developed a reproductive tract size and position score (SPS) system as a reproductive management tool to identify lactating dairy cows with decreased fertility. This system, relying solely on transrectal palpation, considers the size (cervical and uterine) and position of the reproductive tract relative to the pelvis. Cows undergoing pre-breeding exams were identified as having reproductive tracts that were small (SPS1), medium (SPS2), or large (SPS3). Cows designated SPS1 had small and compact uterine horns that rested within the pelvic cavity; SPS2 cows had reproductive tracts that were intermediate in cervical and uterine horn diameter, with longer uterine horns resting partially outside the pelvic cavity; and SPS3 cows had reproductive tracts that were larger and rested mostly outside the pelvic cavity. Cows that were SPS1 had a higher rate of pregnancy per artificial insemination (43.3 ± 3.7%) than cows that were SPS2 (36.9 ± 3.6%) or SPS3 (27.7 ± 4.3%). The percentage of cows with an SPS2 score differed in pregnancies per artificial insemination compared with SPS3 cows. The average days in milk was similar for SPS1, SPS2, and SPS3 cows (104.3 ± 3.5, 98.4 ± 3.4, and 94.7 ± 7.7, respectively). Ultrasound measurements of the uterine horn and cervical diameter, and length measurements of the uterine horns, cervix, and vagina confirmed differences among the SPS groups derived by transrectal palpation. The ease with which transrectal palpation can be used to determine the size and position of the reproductive tract attests to the relevance and usefulness of this scoring system to identify less fertile lactating dairy cows. The ability to do so with ease provides an opportunity to make economically relevant management decisions and maximize reproductive efficiency in a given herd.

Relationships among temperament, acute and chronic cortisol and testosterone concentrations, and breeding soundness during performance testing of Angus bulls.

The aim of this study was to examine relationships among temperament, endocrinology, and reproductive parameters of bulls enrolled in an 84-day performance test. Angus bulls (n = 60) were housed in six pens grouped by age and weight. Pen scores (PS; 1 = docile to 5 = very aggressive) were assigned on Days -1, 27, 55, and 83 of the performance test. On the following day, blood and hair samples were collected, and body weight (BW) and exit velocity (EV) were recorded. Bulls were split into two categories based on; Day -1 PS (PScalm = PS 1 or 2; PSexcitable = PS 3 or 4) and Day 0 EV (EVcalm = slowest 20 bulls; EVexcitable = fastest 20 bulls). Cortisol and testosterone concentrations in serum and hair did not differ (P > 0.10) between PS or EV temperament categories. Sampling day differences (P < 0.01) occurred for serum testosterone, hair cortisol, and hair testosterone concentration; however, serum cortisol concentration did not differ (P > 0.10) over the sampling days. Serum testosterone concentration increased (P < 0.01) from Day 0 to 28, decreased from Day 28 to 56, but Day 84 did not differ from Day 0, 28, or 56. Hair cortisol concentration was greatest (P < 0.01) on Day 0, decreased from Day 28 to 56 but did not differ from Day 56 to 84. Hair testosterone concentration was greatest (P < 0.01) on Day 0 and remained constant from Day 28 to 84. Bulls categorized as PScalm had a greater (P < 0.01) percentage of normal sperm and secondary defects (P < 0.01) when compared with PSexcitable bulls. However, EVcalm bulls had fewer (P < 0.01) primary defects but more (P < 0.01) secondary defects than EVexcitable bulls. In conclusion, bulls exhibited physiological evidence of acclimation during the test as indicated by a reduction in hair cortisol concentration. In addition, the ability of the bulls to acclimate while residing at the testing center may have contributed to little differences observed during the breeding soundness examination portion of the performance test.

Genotype-dependent Metabolic Responses to Semi-Purified High-Sucrose High-Fat Diets in the TALLYHO/Jng vs. C57BL/6 Mouse during the Development of Obesity and Type 2 Diabetes.

Background: The co-epidemic of obesity and type 2 diabetes is associated with increased morbidity and mortality. Genetic factors are highly involved in the development of these diseases, in the form of interactions of multiple genes within obesogenic and diabetogenic environments, such as a high fat diet. The TALLYHO/Jng (TH) mouse is an inbred polygenic model for human obesity and type 2 diabetes. In order to further develop the TH mouse as a clinically relevant model, we investigated diet dependence of obesity and type 2 diabetes in TH mice vs. C57BL/6 (B6) mice. Results: TH and B6 mice were weaned onto a standard rodent chow, semi-purified high-sucrose low-fat (HSLF), or semi-purified high-sucrose high-fat (HSHF) diet and maintained on these diets throughout the study. Despite similar fat contents in HSLF diets and chow, both B6 and TH mice responded to HSLF diets, with increases in adiposity. TH mice, but not B6 mice, exhibited significantly higher adiposity with severely aggravated glucose intolerance and hyperglycemia on HSHF diets compared to the other diets. HSLF diets also advanced diabetes in TH mice compared to chow, but it did not surpass the effects of HSHF diets. The severe glucose intolerance and hyperglycemia in TH mice on both HSLF and HSHF diets were accompanied by significantly reduced Glut4 mRNA levels compared to B6 mice. Conclusions: The present data demonstrate that diets are important modulators of genetic susceptibility to type 2 diabetes and obesity in TH mice. The interplay between heredity and dietary environment in TH mice appears to amplify insulin resistance, contributing to severe glucose intolerance and diabetes.

Hemlock Woolly Adelgid (Hemiptera: Adelgidae) Abundance and Hemlock Canopy Health Numerous Years After Imidacloprid Basal Drench Treatments: Implications for Management Programs.

Hemlock woolly adelgid (Adelges tsugae [Annand]), an invasive insect in the eastern United States, has caused widespread decline of eastern hemlock, Tsuga canadensis (L.) Carriére. Imidacloprid basal drench treatments were assessed 4-7 yr after a single treatment to determine hemlock woolly adelgid population suppression and effects on hemlock canopy health. The effects of sampling site, years post-treatment, and hemlock diameter at breast height (DBH) size classes were evaluated relative to imidacloprid treatment on hemlock woolly adelgid populations and hemlock canopy health characteristics. The influence of hemlock woolly adelgid populations on canopy health characteristics was also assessed. Imidacloprid treatments resulted in low-level hemlock woolly adelgid populations 7 yr post-treatment. Hemlock woolly adelgid was present on more hemlocks 7 yr compared with 4-6 yr post-treatment. Smaller hemlocks, dosed with 0.7 g active ingredient (AI)/2.5 cm DBH, had higher populations of hemlock woolly adelgid than the largest size class, which were treated at twice that dosage. Concentrations of imidacloprid and its olefin metabolite below the LC50 were sufficient for suppression of hemlock woolly adelgid populations, which suggests an additive effect of imidacloprid and olefin that compounds hemlock woolly adelgid mortality over many generations. Hemlock woolly adelgid populations observed in this study were too low to have an observable effect on hemlock canopy health, indicating that application intervals of up to 7 yr may be adequate to protect hemlocks.

Relationships among temperament, behavior, and growth during performance testing of bulls.

Excitable cattle are dangerous to personnel and have reduced individual performance. The aim of this study was to 1) identify objective criteria for evaluating bull temperament and 2) examine relationships among temperament, behavior, and performance of bulls during an 84-d performance test. Angus bulls ( = 60) were reared in 6 pens based on BW and age. Pen scores (PS; 1 = docile and 5 = very aggressive) were assigned on d -1, 27, 55, and 83. Exit velocity (EV), BW, time to exit the chute, and order through the chute were recorded on d 0, 28, 56, and 84. The ADG was calculated for the 84-d test period, and ultrasound data and frame score calculations were recorded on d 84. Dataloggers measured steps taken, lying time, number of lying bouts, and lying bout duration of bulls ( = 27; 3 pens) from d 3 to 28 and d 59 to 84. Bulls with a d -1 PS of 1 or 2 were categorized as calm (PScalm; = 40), whereas bulls with a PS of 3 or 4 were categorized as excitable (PSexcitable; = 20). Bulls were separated into 2 groups based on the bottom 20 EV (EVcalm) and top 20 EV (EVexcitable) on d 0. Mixed model ANOVA (SAS 9.3) was used to compare groups for the two temperament assessment methods, behavior, and growth performance. Mean EV decreased ( < 0.05) by d 84. Total lying time from d 3 to 28 was greater ( < 0.05) for PScalm bulls when compared with PSexcitable bulls. However, total lying time from d 59 to 84 was greater ( < 0.05) for EVexcitable bulls when compared with EVcalm bulls. Regardless of initial contemporary group assignment, all bulls exited the chute slower ( < 0.001) on d 84 than on d 0. The PSexcitable bulls had greater ( < 0.01) frame scores and greater ADG than PScalm bulls. The PSexcitable bulls had more ( < 0.01) backfat than PScalm bulls. However, ribeye area was smaller ( < 0.01) in EVexcitable bulls than EVcalm bulls. Based on these results, bulls appeared to have habituated over the testing period. Additionally, the potential lack of innate temperament variation may have attributed to the little difference seen among the behavioral and performance data. Therefore, temperament should be reassessed within a novel environment with new handlers to differentiate between the bull's true temperament and its ability to habituate.

A polymorphism in XKR4 is significantly associated with serum prolactin concentrations in beef cows grazing tall fescue.

Fescue toxicosis is a common syndrome of poor growth and reproductive performance of beef cattle grazing endophyte-infected tall fescue infected with Lolium arundinaceum Schreb. Together with decreased feed intake, decreased growth rates and tissue necrosis due to vasoconstriction, depressed circulating serum prolactin concentrations are typically observed in cattle afflicted with fescue toxicosis. Polymorphisms within the XK, Kell blood group complex subunit-related family, member 4 (XKR4) gene located on BTA14 have been previously reported to be associated with rump fat thickness, residual feed intake, average daily feed intake and average daily gain in cattle. Associations also have been reported between XKR4 genotype and effectiveness of the dopamine antagonist iloperidone as a treatment of schizophrenia in humans. Domperidone, a related dopamine antagonist, mediates effects of fescue toxicosis on livestock, including restoring depressed concentrations of prolactin. A mixed-breed population of 592 beef cattle grazing endophyte-infected tall fescue was used to examine the association between XKR4 genotype and circulating prolactin concentrations. The SNP rs42646708 was significantly (P = 0.0002) associated with serum prolactin concentrations and explained 2.45% of the phenotypic variation. Effect of genotype at the SNP was tested across five breeds, with significant associations within both Angus (P = 0.0275) and Simmental (P = 0.0224) breeds. These results suggest XKR4 may play a role in mediating the negative effects of fescue toxicosis, and polymorphisms within this gene may be useful markers for selection for genetic resistance to the debilitating effects of endophyte-infected tall fescue.

Heat stress effects on the cumulus cells surrounding the bovine oocyte during maturation: altered matrix metallopeptidase 9 and progesterone production.

When the effects of heat stress are detrimental during maturation, cumulus cells are intimately associated with the oocyte. To determine the extent to which heat stress affects these cells, in this study, transcriptome profiles of the cumulus that surrounded control and heat-stressed oocytes (41 °C during the first 12 h only and then shifted back to 38.5 °C) during in vitro maturation (IVM) were compared using Affymetrix bovine microarrays. The comparison of cumulus-derived profiles revealed a number of transcripts whose levels were increased (n=11) or decreased (n=13) ≥ twofold after heat stress exposure (P<0.01), sufficient to reduce the development of blastocysts by 46.4%. In a separate study, quantitative PCR (qPCR) was used to confirm heat-induced differences in the relative abundances of the transcripts of five different genes (caveolin 1, matrix metallopeptidase 9, FSH receptor, Indian hedgehog homolog, and inducible nitric oxide synthase). Heat stress exposure resulted in >1.7-fold decrease in the protein levels of latent matrix metallopeptidase 9 (proMMP9). Heat-induced reductions in transcript levels were noted at 6 h IVM with reductions in proMMP9 protein levels at 18 h IVM (P=0.0002). Independent of temperature, proMMP9 levels at 24 h IVM were positively correlated with the development rate of blastocysts (R²=0.36; P=0.002). The production of progesterone increased during maturation; heat-induced increases were evident by 12 h IVM (P=0.002). Both MMP9 and progesterone are associated with the developmental competence of the oocyte; thus, it seems plausible for some of the negative consequences of heat stress on the cumulus-oocyte complex to be mediated through heat-induced perturbations occurring in the surrounding cumulus.

Ovulatory follicle dysfunction in lactating dairy cows after treatment with Folltropin-V at the onset of luteolysis.

The objective was to examine growth of the ovulatory follicle after FSH (Folltropin-V; Bioniche Animal Health, Belleville, Ontario, Canada) was given at the onset of induced luteolysis during a synchronization of ovulation protocol. Using GnRH or hCG for inducing ovulation enabled assessing ovulatory follicle responsiveness to an endogenous versus exogenous surge of LH activity. At 8 to 10 days after estrus (synchronized estrus = Day 0), lactating dairy cows received an Eazi-Breed CIDR (Pfizer Animal Health) plus 100 µg GnRH. After 7 days, controlled internal drug release devices (CIDRs) were removed, cows were given 500 µg cloprostenol, and then randomly allocated to receive 80 mg Folltropin-V (FSH; N = 19) or 4 mL sterile saline (SAL; N = 16). After 49 hours, FSH and SAL cows were randomly allocated to receive 100 µg GnRH or 3000 IU hCG. Five cows ovulated 30 to 42 hours (38.4 ± 1.2 hours) after FSH treatment. In the remaining FSH (N = 14) or SAL (N = 16) cows, ovulatory follicle size was similar at CIDR removal (14.5 ± 0.6 and 14.7 ± 0.6 mm, respectively; P = 0.85) and when GnRH/hCG was given (16.6 ± 0.6 and 17.7 ± 0.6 mm, respectively; P = 0.23). Estradiol-17ß concentrations were lower in FSH cows at 36 and 49 hours after CIDR removal (FSH by time interaction, P < 0.005). After GnRH or hCG treatment, four FSH cows failed to ovulate. In cows exhibiting ovulation, the last recorded size of the ovulatory follicle was not influenced by FSH (18.1 ± 0.9 and 17.5 ± 0.6 mm for FSH and SAL, respectively; P = 0.59) or hormonal induction approach (18.4 ± 0.9 and 17.2 ± 0.7 mm for GnRH and hCG, respectively; P = 0.29). The interval from onset of luteolysis to ovulation and pharmaceutical induction to ovulation was shorter in FSH cows given GnRH (FSH by pharmaceutical inducer [GnRH vs. hCG] interaction; P = 0.01). Cows receiving GnRH had an LH surge; hCG-treated cows did not. Maximum LH concentrations were greater (P < 0.04) in SAL versus FSH cows after GnRH treatment (10.9 ± 1.2 vs. 6.7 ± 1.4 ng/mL, respectively). In three FSH cows failing to ovulate after GnRH treatment, the maximum LH concentration was <4 ng/mL. When analyzed from GnRH treatment, average time to LH maximum concentration was similar (P = 0.50) to values obtained in cows receiving FSH and GnRH and SAL and GnRH (1.7 ± 0.2 vs. 1.9 ± 0.1 hours, respectively). Interval to maximum hCG concentrations was shorter (P = 0.02) for cows receiving SAL versus FSH (8.0 ± 0.8 and 10.0 ± 0.8 hours for SAL and FSH, respectively). Ovulatory dysfunction of this magnitude highlighted the lack of suitability of Folltropin-V at a dose of 80 mg at the time of induction of luteolysis in fixed timed AI protocols.

Evaluation of regional limb perfusion with amikacin using the saphenous, cephalic, and palmar digital veins in standing horses.

Previous studies have shown that regional limb perfusion (RLP) using the palmar digital (PD) vein delivers therapeutic concentration of amikacin to the distal limb. Our hypothesis was that using the cephalic and saphenous veins for RLP will enable delivery of therapeutic concentrations of amikacin to the distal limb. Nineteen healthy horses participated in the study. The cephalic, saphenous, or PD vein was used to perfuse the limb with amikacin. Two grams of amikacin was used for RLP using the saphenous and the cephalic veins, and one gram was used in the PD vein. Synovial samples were collected from the metacarpo-/metatarsophalangeal (MCP/MTP) joint, and blood samples were collected from the jugular vein. Maximum concentration (Cmax) of amikacin in the MCP/MTP joint using the cephalic and the saphenous vein was 277 and 363 mg/L, respectively. The amikacin concentrations achieved in the synovial fluid of the MCP/MTP joint in the current study were between 69 and 91 times the minimally inhibitory concentration of common susceptible bacterial pathogens causing orthopedic infections in horses. To conclude, this study shows that use of the proximal veins for RLP to treat distal limb infections is a viable alternative to using the palmar or plantar digital vein.

Evaluation of regional limb perfusion with erythromycin using the saphenous, cephalic, or palmar digital veins in standing horses.

There are no reported studies evaluating the use of erythromycin for regional limb perfusion (RLP) in horses. Our hypothesis was that using the cephalic and saphenous veins for RLP will enable delivery of therapeutic concentrations of erythromycin to the distal limb. Nineteen healthy horses participated in the study. The cephalic, saphenous or palmar digital (PD) vein was used to perfuse the limb with erythromycin. Synovial samples were collected from the metacarpo/metatarso-phalangeal (MCP/MTP) joint and blood samples were collected from the jugular vein. Maximum concentration (C(max)) of erythromycin in the MCP joint using the cephalic vein was 113 mg/L. The Cmax of erythromycin in the MTP joint using the saphenous vein was 38 mg/L. Erythromycin administered using the PD vein was not detectable in the MCP/MTP joint of four of six horses. Concentrations of erythromycin achieved in the synovial fluid of the MCP/MTP joint were between 152 and 452 times the minimal inhibitory concentration (MIC) for Rhodococcus equi (R. equi). In conclusion, the results indicate that when using the saphenous or cephalic veins for RLP, therapeutic concentrations of erythromycin in the MCP/MTP joint can be consistently reached [corrected].

Disparate consequences of heat stress exposure during meiotic maturation: embryo development after chemical activation vs fertilization of bovine oocytes.

Consequences of heat stress exposure during the first 12 h of meiotic maturation differed depending on how and when bovine oocytes were activated. If heat-stressed oocytes underwent IVF at ~24 h, blastocyst development was less than for respective controls and similar to that obtained for nonheat-stressed oocytes undergoing IVF at 30 h (i.e. slightly aged). In contrast, if heat-stressed oocytes underwent chemical activation with ionomycin/6-dimethylaminopurine at 24 h, blastocyst development was not only higher than respective controls, but also equivalent to development obtained after activation of nonheat-stressed oocytes at 30 h. Developmental differences in chemically activated vs IVF-derived embryos were not related to fertilization failure or gross alterations in cytoskeletal components. Rather, ionomycin-induced calcium release and MAP kinase activity were less in heat-stressed oocytes. While underlying mechanisms are multifactorial, ability to obtain equivalent or higher development after parthenogenetic activation demonstrates that oocytes experiencing heat stress during the first 12 h of meiotic maturation have the necessary components to develop to the blastocyst stage, but fail to do so after fertilization.

Evaluation of indwelling intravenous catheters for the regional perfusion of the limbs of horses.

The feasibility of maintaining indwelling intravenous catheters in the saphenous, cephalic or palmar digital vein of horses for seven days to infuse antimicrobial drugs was investigated in 18 horses. The horses were randomly assigned to six groups according to the vein catheterised and whether they received amikacin or erythromycin. None of the catheters was replaced more than once, and 11 of the 18 catheters remained patent for all seven days. Neither the drug administered nor the vein catheterised significantly affected the survival of the catheter. In all but three cases, complications, including local inflammation, lameness, thrombophlebitis and one severe tissue reaction to erythromycin, resolved during the seven days.

Developmental competence of bovine embryos from heat-stressed ova.

Because multiple ovulation embryo transfer procedures are occasionally performed in cows experiencing heat stress, the goal of this study was to assess the developmental competence of otherwise morphologically normal embryos from heat-stressed ova. To this end, the ability of compact morulae from heat-stressed and non-heat-stressed bovine ova to undergo blastocyst development after culture at 38.5 or 41.0 degrees C was examined. It was hypothesized that heat-induced perturbations in the ooplasm carry over to increase the susceptibility of the preattachment embryo to heat stress. Initially, ova were matured at 38.5 or 41.0 degrees C. The consequences of heat stress did not include altered cleavage, but did reduce the proportion of 8- to 16-cell-stage embryos (55.3 vs. 50.6%; SEM +/- 1.9). Although proportionately fewer, compact morulae from heat-stressed ova were equivalent in quality to those from non-heat-stressed ova (2.1 and 2.1; SEM = 0.04). Culture of compact morulae from non-heat-stressed ova at 41.0 degrees C did not affect blastocyst development (71.9 and 71.5%; SEM = 3.0). Furthermore, the development of compact morulae from heat-stressed ova was similar to that of non-heat-stressed ova after culture at 38.5 degrees C (68.2 vs. 71.9 and 71.5%; SEM = 3.0). However, blastocyst development was reduced when compact morulae from heat-stressed ova were cultured at 41.0 degrees C (62.3 vs. 71.9, 71.5 and 68.2; SEM = 3.1). In summary, reduced compaction rates of heat-stressed ova explained in part why fewer develop to the blastocyst stage after fertilization. The thermolability of the few embryos that develop from otherwise developmentally challenged ova emphasizes the importance of minimizing exposure to stressor(s) during oocyte maturation.

In vitro effects of hydrochloric and lactic acids on bioelectric properties of equine gastric squamous mucosa.

REASONS FOR PERFORMING STUDY: Volatile fatty acids, byproducts of carbohydrate fermentation by resident bacteria, have been implicated in causing nonglandular (NG) gastric ulcers. Lactic acid (LA), also produced by stomach bacteria, may cause gastric ulcers when exposed to the equine NG mucosa. OBJECTIVES: To investigate the in vitro effects of LA on equine NG mucosa bioelectric properties, sodium transport and tissue resistance. METHODS: Gastric tissues obtained from 13 mature horses were studied in Ussing chambers. Short-circuit current (Isc) and potential difference (PD) were measured, and electrical resistance (R) and conductance (G) calculated for tissues after addition of HCl and LA (5, 10, 20 and 40 mmol/l) in normal Ringer's solution (NRS). RESULTS: Mucosa exposed to HCl or LA (5, 10 and 20 mmol/l) in NRS (pH 1.5 and to a lesser extent pH 4.0) had a significant decrease in Isc and PD. Mucosa exposed to a high concentration of LA (40 mmol/l) in NRS (LRS) at pH 1.5 showed an increased G, but this increase was not significant. Values returned to baseline after solutions were returned to pH 7.0. Histological changes were consistent with HCl-induced (pH <4.0) acid damage. CONCLUSIONS: HCl induced alteration in bioelectric properties of equine NG mucosa whereas addition of LRS did not, other than those changes seen with HCl alone.

Effects of a concentrated lidocaine solution on the acute phase stress response to dehorning in dairy calves.

The objective of this study was to more fully define the surgical stress response to dehorning by heat cauterization in dairy calves by measuring behavioral, hormonal, inflammatory, and immunological markers of stress and to determine whether a nerve block of the surgical site with a concentrated solution of lidocaine (5%) reduces the degree of stress. Thirty-two 10- to 12-wk-old female Holstein calves were randomly allotted to 1 of 4 treatments: 5% lidocaine followed by dehorning, 2% lidocaine followed by dehorning, saline followed by dehorning, or 5% lidocaine followed by sham dehorning. Plasma cortisol concentration was measured in blood samples collected via a jugular catheter at -0.5, 0, 0.5, 1, 1.5, 2, 2.5, 3, 3.5, 4, 6, 9, 12, 24, 48, and 72 h. Various other blood constituents were measured in samples collected at -0.5, 12, 24, 48, and 72 h. Feeding, drinking, scratching, grooming, rubbing, licking, and inactivity behaviors were observed in the standing and recumbent positions using a 10-min scan sampling method analyzed on a time period and daily basis for 72 h following the dehorning procedure. The frequency of vocalization, kicking, and lying in the chute during the dehorning procedure were also assessed. The overall plasma cortisol concentrations were higher in calves subjected to dehorning than in control calves. Compared with the control group, the saline-treated calves had a higher cortisol concentration at 30 and 60 min postdehorning. Plasma cortisol concentrations were higher in all groups at 30 min postdehorning than at other sampling times. The percentage of circulating neutrophils and the neutrophil:lymphocyte ratio were increased in the saline and 2% lidocaine group. Total plasma protein, fibrinogen, and alpha1-acid glycoprotein concentrations were similar among treatments. The behavioral response to dehorning, as manifested by kicking while in the chute, was greater in the saline and 2% lidocaine group than in the control or 5% lidocaine treatment groups. In the postdehorning period, the percentage of time calves spent performing various maintenance behaviors did not differ among treatments. Thus, injection of 5% lidocaine may not provide any added comfort after the dehorning but may decrease the overall stress response during the procedure.

Early in vitro fertilization improves development of bovine ova heat stressed during in vitro maturation.

The objectives were to examine the development of embryos derived from control (38.5 degrees C) or heat-stressed ova [41.0 degrees C during the first 12 h of in vitro maturation (hIVM)] when in vitro fertilization (IVF) was performed at 16, 18, 20, 24, or 30 hIVM. Effects of heat stress in compromising ovum development depended on when IVF was performed (in vitro maturation temperature x IVF time interaction). When IVF was performed at 24 or 30 hIVM, fewer heat-stressed ova developed to the blastocyst stage compared with the respective controls. In contrast, when IVF was performed at 16, 18, or 20 hIVM, more heat-stressed ova developed to the blastocyst stage compared with the respective controls. Performing IVF earlier than usual was beneficial, because the ability of heat-stressed ova to develop to the blastocyst stage was improved when IVF was performed at 18 or 20 vs. 24 hIVM. Blastocyst stage and quality were equivalent to non-heat-stressed controls regardless of IVF time. Control ova undergoing IVF at 20, 24, 30, or 32 hIVM and heat-stressed ova undergoing IVF at 16, 18, 20, or 24 hIVM were compared for blastocyst development by multisource regression. Although linear and quadratic slopes were similar, heat stress reduced the peak and shifted the developmental response of ova by 7.3 h. In other words, obtaining optimal blastocyst development from heat-stressed ova would depend on performing IVF at 19.5 hIVM compared with 26.7 hIVM for non-heat-stressed controls. Heat-induced reductions in peak blastocyst development significantly reduced the window of time available to perform IVF and obtain > or = 20% blastocyst development. In summary, results support an effect of heat stress to hasten developmentally important events during oocyte maturation. The inability of earlier IVF to fully restore the development of heat-stressed ova to that of non-heat-stressed controls highlights the importance of further study.

Manipulation of thyroid status and/or GH injection alters hepatic gene expression in the juvenile chicken.

Both thyroid hormone (T3) and growth hormone (GH) are important regulators of somatic growth in birds and mammals. Although T3-mediated gene transcription is well known, the molecular basis of T3 interaction with GH on growth and development of birds remains unknown. In earlier studies, we discovered that exogenous GH alone increased accumulation of visceral fat in young chickens, while the combination of GH injections and dietary T3 worked synergistically to deplete body fat. In the present study, cDNA microarray and quantitative RT-PCR analyses enabled us to examine hepatic gene expression in young chickens after chronic manipulation of thyroid status and GH injection alone or in combination with T3. Thyroid status modulates expression of common and unique sets of genes involved in a wide range of molecular functions (i.e., energy metabolism, storage and transport, signal transduction, protein turnover and drug detoxification). Hepatic expression of 35 genes was altered by hypothyroidism (e.g., ADFP, ANGPTL3, GSTalpha, CAT, PPARG, HMGCL, GHR, IGF1, STAT3, THRSPalpha), whereas hyperthyroidism affected expression of another cluster of 13 genes (e.g., IGFBP1, KHK, LDHB, BAIA2L1, SULT1B, TRIAD3). Several genes were identified which have not been previously ascribed as T3 responsive (e.g., DEFB9, EPS8L2, ARHGAP1, LASS2, INHBC). Exogenous GH altered expression of 17 genes (e.g., CCAR1, CYP2C45, GYS2, ENOB, HK1, FABP1, SQLE, SOCS2, UPG2). The T3+GH treatment depleted the greatest amount of body fat, where 34 differentially expressed genes were unique to this group (e.g., C/EBP, CDC42EP1, SYDE2, PCK2, PIK4CA, TH1L, GPT2, BHMT). The marked reduction in body fat brought about by the T3+GH synergism could involve modulation of hormone signaling via altered activity of the Ras superfamily of molecular switches, which control diverse biological processes. In conclusion, this study provides the first global analysis of endocrine (T3 and GH) regulation of hepatic gene transcription in the chicken.

Temporal relationships between plasma cortisol, corticosteroid-binding globulin (CBG), and the free cortisol index (FCI) in pigs in response to adrenal stimulation or suppression.

The objective of this study was to document changes in plasma concentrations of total cortisol, porcine corticosteroid-binding globulin (pCBG), and the free cortisol index (FCI) in pigs over a 6-h period in response to adrenal stimulation or suppression. Twenty-four 8-week old pigs allotted in equal numbers were administered ACTH, dexamethasone or saline, and blood samples were collected every 15 min via an indwelling jugular catheter for 1 h prior to and 5 h following treatment. Total plasma cortisol increased in ACTH-treated pigs and decreased in dexamethasone-treated pigs within 0.25 and 0.5 h, respectively. In contrast, pCBG concentration was altered in an inverse fashion subsequent to the changes exhibited in total cortisol. FCI reflected the changes observed in total cortisol. These results further document the negative relationship that exists between circulating concentrations of plasma cortisol and pCBG, and illustrate that this association exists under conditions of acute stress in the pig.