Hair cortisol levels of lactating dairy cows in cold‐ and warm‐temperate regions in Japan

We compared the hair cortisol levels of lactating dairy cows in a cold‐ and a warm‐temperate region out of four climatic zones in Japan. We simultaneously investigated the effects of calving number, lactation period and month of hair sampling. Hair of nine Holstein lactating cows chosen from each region (i.e. 18 cows per sampling) was sampled in March, June, September and December. Number of calvings (1, 2, ≥3) and lactation duration (early: <100, middle: 101–200, and late: >201 days) were balanced between regions. Cortisol was extracted from hair by methanol, and its level was determined with a cortisol immunoassay kit. A multi‐way analysis of variance revealed that the effects of month of hair sampling (P < 0.001) and its combination with region (P < 0.05) were significant. In a multiple comparison test, significant differences (P < 0.01) in hair cortisol level (pg/mg of hair) were found between June (13.0 ± 1.0) and the other 3 months, and between September (1.6 ± 0.2) and December (4.5 ± 0.3). The rise in cortisol level from March to June was more intense in the cold‐temperate region. These results demonstrate the necessity of considering seasonal variations in each climatic region when we use hair cortisol level as an indicator of stress.     

Comparison of productive and reproductive performance and hair cortisol levels between Brown Swiss cross‐bred and Holstein cows housed in the same barn

The productive and reproductive characteristics of Brown Swiss (B) cross‐bred cows were investigated by comparing with those of Holstein (H) cows housed in the same barn. Additionally, their hair cortisol levels were analyzed to evaluate the extent of stress experienced during dry and lactation periods. B cross‐bred cows had lower milk yields and higher milk fat rates than H cows. Reproductive records showed that days from parturition to first artificial insemination (AI) in B cross‐bred (n = 16) and H (n = 27) cows were not significantly different, but conception rate at first AI of B cross‐bred cows tended to be higher than that of H cows. Percentage of B cross‐bred cows that resumed ovarian cyclic activity within 45 days after parturition was higher than that of H cows (6/6 (100%) and 5/11 (45.5%), P < 0.05), and B cross‐bred cows had higher body condition scores at that time. Hair cortisol level at 60 to 90 days after parturition in H cows increased significantly compared with in the dry period, and it was higher than that of B cows during the same period. These results suggest that B cross‐bred cows experience less metabolic stress during early lactation, which may result in earlier resumption of reproductive function.     

Effect of growth and parturition on hair cortisol in Holstein cattle

The effect of growth and parturition on hair cortisol concentrations of cattle was investigated. Plasma, saliva, and hair (black and white from the shoulders and hip) samples were collected from calves at 6 and 24 weeks old and from dairy cattle at the dry (1 and 2 months prepartum) and lactation (10, 50, 150, and 250 days postpartum) periods. Plasma and saliva cortisol concentrations were lower in 24-week-old calves than those of 6-week-old calves, and hair cortisol concentrations decreased regardless of color and position. In 6-week-old calves, hair cortisol concentrations differed between sampling positions, but this difference was not observed in 24-week-old calves. Plasma and saliva cortisol concentrations increased before parturition until 10 days postpartum then decreased until 50 days postpartum. The same trend was observed in the cortisol concentrations of white hair. Contrarily, cortisol concentrations in black hair remained unchanged and was lower than that in white hair. Hair cortisol concentration can vary greatly depending on the location on the body, hair color, cattle age, or parturition. When this method is used, all of the above factors must be considered.     

Hair cortisol concentration in pre‐ and postpartum dairy cows,and its association with body condition,hock health,and reproductive status

The objective of this study was to investigate the profiles of hair cortisol concentrations as an index of chronic stress in dairy cows in association with their health, nutrition, and reproductive parameters. For 25 Holstein dairy cows, hair was collected from the tail switch ?19.2 ± 11.4, 44.8 ± 11.9, 103.0 ± 9.9, and 168.0 ± 9.7 days postpartum (L0, L1, L2, and L3, respectively). Body condition scores were negatively correlated with hair cortisol concentrations (r = ?0.255), and hock health scores were positively correlated with hair cortisol concentrations (r = 0.236, p < 0.05). Hair cortisol concentrations during the postpartum period showed different patterns according to the time of first artificial insemination (AI) and fertility. Cows that were submitted to first AI by 86 days postpartum showed a peak hair cortisol concentration at L1, whereas cows with delayed first AI had a peak at L2. The hair cortisol concentrations of subfertile (≥168 days) cows were significantly higher at L1 and L2 compared with L0, whereas hair cortisol concentrations of fertile cows (<168 days) were not different among the sampling times. These results indicate that cows with health problems appear to experience greater chronic stress, which may impair their reproductive function.     

Effects of adrenocorticotropic hormone challenge and age on hair cortisol concentrations in dairy cattle

Dairy cattle suffer stress from management and production; contemporary farming tries to improve animal welfare and reduce stress. Therefore, the assessment of long-term hypothalamic-pituitary-adrenal function using non-invasive techniques is useful. The aims in this study were: to measure cortisol concentration in cow and calves hair by radioimmunoassay (RIA), to test cortisol accumulation in bovine hair after adrenocorticotropic hormone (ACTH) challenges, and determine the influence of hair color on cortisol concentrations. Fifteen Holstein heifers were allotted to 3 groups (n = 5 each): in control group (C), just the hair was sampled; in the saline solution group (SS), IV saline solution was administered on days 0, 7, and 14; and the ACTH group was challenged 3 times with ACTH (0.15 UI per kg of body weight) on days 0, 7, and 14. Serum samples from the SS and ACTH groups were obtained 0, 60 and 90 min post-injection. Serum cortisol concentration was greater 60 and 90 min after injection with ACTH. Hair was clipped on days 0, 14, 28, and 44. Hair cortisol was methanol extracted and measured by RIA. Hair cortisol was preserved for 11 mo. Hair cortisol concentrations in the ACTH group were greater than in the saline and control groups on days 14 and 28, but not on day 44. Concentrations were greater in calves than in cows and greater in white hair than in black hair. Cortisol accumulated in bovine hair after ACTH challenges, but the concentration was affected by both age and hair color. If hair color effects are taken into account, assessing cortisol concentration in hair is a potentially useful non-invasive method for assessing stress in cattle.     

Evidence of a major gene influencing hair length and heat tolerance in Bos taurus cattle

Evidence was found that supports the existence of a major gene (designated as the slick hair gene), dominant in mode of inheritance, that is responsible for producing a very short, sleek hair coat. Cattle with slick hair were observed to maintain lower rectal temperatures (RT). The gene is found in Senepol cattle and criollo (Spanish origin) breeds in Central and South America. This gene is also found in a Venezuelan composite breed, the Carora, formed from the Brown Swiss and a Venezuelan criollo breed. Two sets of backcross matings of normal-haired sire breeds to Senepol crossbred dams assumed to be heterozygous for the slick hair gene resulted in ratios of slick to normal-haired progeny that did not significantly differ from 1:1. Data from Carora x Holstein crossbred cows in Venezuela also support the concept of a major gene that is responsible for the slick hair coat of the Carora breed. Cows that were 75% Holstein: 25% Carora in breed composition segregated with a ratio that did not differ from 1:1, as would be expected from a backcross matinginvolving a dominant gene. The effect of the slick hair gene on RT depended on the degree of heat stress and appeared to be affected by age and/or lactation status. The decreased RT observed for slick-haired crossbred calves compared to normal-haired contemporaries ranged from 0.18 to 0.4 degrees C. An even larger decrease in RT (0.61 degrees C; P < 0.01) was observed in lactating Carora x Holstein F1 crossbred cows, even though it did not appear that these cows were under severe heat stress. The improved thermotolerance of crossbred calves due to their slick hair coats did not result in increased weaning weights, possibly because both the slick and normal-haired calves were being nursed by slick-haired dams. There were indications that the slick-haired calves grew faster immediately following weaning and that their growth during the cooler months of the year was not compromised significantly by their reduced quantity of hair. In the Carora x Holstein crossbred cows there was a positive effect of slick hair on milk yield under dry, tropical conditions.     

KIT基因多态性与荷斯坦牛着色性状的关联分析

牛的被毛颜色是重要的外貌性状。本研究采集北京地区401头荷斯坦母牛血样及21头公牛冻精样品．提取基因组DNA,选取牛6号染色体上KIT基因的4个SNPs位点,利用飞行时间质谱技术检测基因型。通过数码相机照相并进行图像分析计算牛的被毛黑白比例,记录乳头颜色。将基因型和表型数据进行关联分析,结果显示KIT基因的G72792875T住点对被毛比例有显著影响（P〈0．05）,推测KIT基因可能与荷斯坦牛的着色性状的形成相关。     

Transcriptome sequencing of black and white hair follicles in the giant panda

With the completion of the draft assembly of the giant panda genome sequence, RNA sequencing technology has been widely used in genetic research on giant pandas. We used RNA-seq to examine black and white hair follicle samples from adult pandas. By comparison with the giant panda genome, 75 963 SNP loci were labeled, 2426 differentially expressed genes (DEGs) were identified, and 2029 new genes were discovered, among which 631 were functionally annotated. A cluster analysis of the DEGs showed that they were mainly related to the Wnt signaling pathway, ECM–receptor interaction, the p53 signaling pathway, and ribosome processing. The enrichment results showed that there were significant differences in the regulatory networks of hair follicles with different colors during the transitional stage of hair follicle resting growth, which may play a regulatory role in melanin synthesis during growth. In conclusion, our results provide new insights and more data support for research on the color formation in giant pandas.     

Heat stress decreased hair follicle population in rex rabbits

The aim of this study was conducted to investigate the effect of heat stress on the hair follicle population and related signalling pathways in rex rabbits. Forty‐eight rabbits were randomly divided into two groups: one group in a high ambient environment (32 ± 2°C, heat stress) and the other group with normal temperature (20 ± 2°C, control). The results show that heat stress decreased the body weight gain and feed conversion rate, rabbit hair length and hair follicle density (p < 0.05). Besides, heat stress suppressed the gene expression of noggin, insulin‐like growth factor 1 (IGF‐1) and IGF‐1 receptor and protein expression of phosphorylated mechanistic target of rapamycin (mTOR) in rabbit skin (p < 0.05), while stimulated significantly the gene expression of bone morphogenetic protein 2 (BMP2) and BMP4 (p < 0.05). Heat exposure did not alter significantly the gene expression of alkaline phosphatase, versican and hepatocyte growth factor compared with the control (p > 0.05). In conclusion, noggin‐BMP, IGF‐1 and mTOR signalling pathways may be associated with the process of heat stress‐repressing hair follicle development.     

Hair cortisol as a marker of hypothalamic-pituitary-adrenal axis activation in Friesian dairy cows clinically or physiologically compromised

This study was designed to assess hypothalamic-pituitary-adrenal (HPA) axis activation in lactating cows clinically or physiologically compromised. Cortisol was measured in the hair of 475 multiparous cows of Italian Friesian breed. The cows were classified into Groups A (n=218) and B (n=257). Group A included cows clinically compromised that had recently suffered a disease (e.g. metritis, laminitis, mastitis) or physiologically compromised that calved 1 month prior to hair sampling. Group B were clinically normal lactating cows at least 60 days after calving with somatic cell counts of less than 200,000 cells/ml in milk. Animals in Group A showed higher hair cortisol levels than those in Group B (P<0.01). Significant positive correlation (rpb=+0.352, P<0.001) was detected between hair cortisol and cows clinically or physiologically compromised. The area under the receiver operating characteristic curve was 0.701 (SE 0.024) and a cut-off value of 4.17 pg/mg was found to discriminate between groups A and B with a sensitivity of 62.4% and specificity of 69.3%. The hair cortisol test emerged as useful to determine HPA activation and to enhance current understanding on the involvement of this axis in clinically or physiologically compromised cows.     

Feed handling of lactating crossbred cows maintained in a semi‐arid region during the hot season: physiological parameters,ingestive behavior and performance

The effects of time of feed delivery (14.00 hours; 14.00 and 20.00 hours; 16.00 and 18.00 hours) on the physiological parameters, ingestive behavior, nutrient intake and production of lactating cows maintained in a semi‐arid region during the hot season were evaluated. Regardless of treatment, all animals received the first feeding supply at 06.00 hours. Eight cows with an average body weight of 600 kg, average milk yield of 20 kg/day and 80 days in milk were utilized. The rectal temperature, respiratory rate and sweating rate were not affected (P > 0.05), with average values of 38.5°C, 53.8 movements/min and 104 g/m²/h respectively. There was no effect (P > 0.05) on the eating time (314 min/day), ruminating time (564 min/day), drinking time (61 min/day) and idle time (502 min/day). Similarly, the intake of nutrients and performance of lactating cows were not affected (P > 0.05), with average dry matter intake of 19.8 kg/day, 4% fat‐corrected milk of 20.6 kg/day and milk fat concentration of 4.03 g/100 g. Since the behavior and performance have not altered, any times of feed delivery evaluated could be used to crossbred Holstein × Zebu cows maintained on a feedlot in semi‐arid regions during the hot season.     

A study of relative horn fly,Haematobia irritans (Diptera: Muscidae), abundance on Holstein steers and steers of two Holstein crosses

A study was performed to determine if the number of horn fly (Haematobia irritans) adults differ significantly on Holstein (black and white coat color), Holstein x Holstein Friesian (black and white coat color) and Holstein x Jersey (black coat color) steers, 10-12 months old at the onset of the study. All steers were run together on lucerne paddocks and the number of flies counted at 30-day intervals from September 2000 to August 2001. No significant differences (P > 0.05, test of Kruskal-Wallis) were found in fly numbers, even in the period April-May 2001 when the infestation reached its peak. We were unable to demonstrate that coat color influenced horn fly abundance in the present study. It appears that none of the biotypes evaluated had any advantage for natural control of H. irritans.     

Skin and coat traits in sheep in Brazil and their relation with heat tolerance

The objective of this study was to evaluate and compare physical parameters in groups of sheep with different phenotypic characteristics in the Centre-west region of Brazil. Five groups of sheep, with nine animals per group, were selected, three groups of Santa Inês animals with different coat colours (white, brown and black), one group with crossbred animals (Santa Inês × Bergamasca) and one group with animals of the Bergamasca breed. The following traits were evaluated: coat thickness, number and length of hair, pigmentation level in the coat and the skin as well as the percentage area of sweats glands in the skin tissue, carried out by histological analysis. The number of hairs and the area of sweats glands were not significantly different between the evaluated groups. The Bergamasca breed showed low pigmentation of the skin and long hairs. The levels of pigmentation of the hair and of the skin were highly correlated. Between the Santa Inês groups, the group with white hair showed the better parameters for heat adaptation, while the brown hair group showed the lower heat adaptation when compared with another hair breed groups.     

Measuring cortisol in hair and saliva from dogs: coat color and pigment differences

Cortisol concentrations are frequently measured from a variety of sources including blood, saliva, urine, and feces to quantify stress in dogs. However, a need still exists for less intrusive collection methods in domestic animals and for more efficient means of measuring basal cortisol. The objectives of the present study were to minimize restraint for saliva sampling, to validate hair for basal cortisol measurement in dogs, and to determine concentrations of cortisol within the hair shaft and in relation to hair color. Using food luring, 79% of dogs required no restraint for saliva collection. Salivary and hair cortisol concentrations were positively correlated (P = 0.001), thus validating hair as a medium for basal cortisol quantification. Black dogs had less cortisol than nonblack dogs (P = 0.039) in hair, but not saliva. Across dogs, the average amount of cortisol did not differ between proximal and distal hair sections (P = 0.348). However, for 7 of the 9 dogs, more cortisol was present in the distal portions of the hair. We observed a difference in cortisol concentrations among hairs of different colors from individual dogs (P = 0.001). From the same 7 × 7 cm ischiatic patch from the same dog, black (eumelanin) hairs were consistently lower in cortisol than yellow (pheomelanin) hairs, and cortisol concentrations of agouti hairs were intermediate. This is the first evidence that hair of different colors might sequester cortisol differently.     

The effect of high ambient temperature on Ca,P and Mg balance and bone turnover in high‐yielding dairy cows

We investigated the effect of heat stress on Ca, P and Mg balance and bone turnover in lactating cows. In a 2 × 2 crossover design, four multiparous lactating Holstein cows were kept in a chamber and subjected to a constant moderate (18°C) ambient temperature (MT) or high (28°C) ambient temperature (HT). The cows were fed total mixed ration (Ca, 0.7%; P, 0.4%; Mg, 0.2%) ad libitum. The milk yield under HT (35.4 kg/day) tended to be lower (P < 0.10) than that under MT (43.2 kg/day). The concentrations of milk P (P < 0.05) and Mg (P < 0.01) were significantly lower under HT than MT. The Ca, P and Mg intake (P < 0.10); Ca (P < 0.10), P, and Mg (P < 0.05) secretion into milk; and Ca (P < 0.05), P (P < 0.01), and Mg (P < 0.05) absorption in the intestine were lower under HT than MT. The plasma osteocalcin, a marker of bone turnover, was significantly lower (P < 0.05) under HT than MT. Heat stress did not affect plasma C‐telopeptide of collagen type I, a bone resorption marker, and plasma parathyroid hormone concentration.     

Efficacy of conventional and extended intra-mammary treatment of persistent sub-clinical mastitis with cefquinome in lactating dairy cows

The objectives of the present study were to evaluate the efficacy of intra-mammary-administered cefquinome for the treatment of sub-clinical mastitis in lactating dairy cows and to determine if extended therapy would enhance treatment efficacy. Seventy-three Holstein dairy cows from a single farm with 150 infected quarters were enrolled in the study. Infected cows were allocated randomly to one of three treatment regimens: (1) conventional (standard) regimen: 75 mg of cefquinome administered three times at 16-h intervals (25 infected cows, 52 intra-mammary infections (IMI)), (2) extended regimen: 75 mg of cefquinome administered six times at 16-h intervals (26 infected cows, 58 IMI) and (3) negative untreated control group (22 cows, 40 IMI). Most IMI were caused by coagulase-negative staphylococci, streptococci other than Streptococcus agalactiae and coliforms. The overall bacteriological cure (BC) rates for sub-clinical IMI were 84.61%, 91.37% and 20% for the conventional, extended and the control groups, respectively, indicating a higher BC rate for the treated groups than the control group (P < 0.001). Significant differences in somatic cell count (SCC) were detected between the treated versus the control group (P < 0.001). No differences, concerning the BC rate or SCC, were observed between the extended and the conventional groups. Although fat and protein percentages increased in the treated groups, there were no significant differences in post-treatment milk production between the groups. Results of this study indicate that cefquinome therapy was effective in reducing SCC and eliminating sub-clinical IMI in lactating dairy cows, but extended therapy did not enhance treatment efficacy.     

Exploration of factors determining milk production by Holstein cows raised on a dairy farm in a temperate climate area

In this study, we examined factors that affected milk production by cows raised in a temperate climate area. We conducted this study on a large dairy farm containing approximately 2000 Holstein cows, located in a temperate climate area. We collected 7803 calving records for 4069 cows from 2012 to 2016. We then assessed the effect of hot weather on milk yield by examining three climate factors: season, maximum temperature (MAX), and the temperature and humidity index (THI). We found that increases in heat stress caused linear decreases in milk yield (P?<?0.05). Additionally, the effects of the three climate factors on milk yield varied depending on cow parity and days open (P?<?0.05). Thus, management procedures should consider cow parity and lactating stage to minimize the negative effects of heat stress on milk production. We also found that the lowest Akaike information criterion value was obtained in our model when using THI for 305-day milk yield. This suggests that THI is a more accurate variable for evaluating heat stress than MAX or season.

     

Effects of amino acids infused into the vein on ghrelin‐induced GH,insulin and glucagon secretion in lactating cows

To investigate the effects of amino acids on ghrelin‐induced growth hormone (GH), insulin and glucagon secretion in lactating dairy cattle, six Holstein cows were randomly assigned to two infusion treatments in a cross‐over design. Mixture solution of amino acids (AMI) or saline (CON) was continuously infused into the left side jugular vein via catheter for 4 h. At 2 h after the start of infusion, synthetic bovine ghrelin was single injected into the right side jugular vein through the catheter. Ghrelin injection immediately increased plasma GH, glucose and non‐esterified fatty acids (P < 0.05) with no difference between both treatments. Additionally, plasma insulin and glucagon concentrations were increased by ghrelin injection in both treatments. The peak value of plasma insulin concentration was greater in AMI compared with CON (P < 0.05). Plasma glucagon concentration showed no difference in the peak value reached at 5 min between both treatments, and then the plasma levels in AMI compared with CON showed sustained higher values (P < 0.05). After plasma glucose concentration reached the peak, the decline was greater in AMI compared with CON (P < 0.05). These results showed that the increased plasma amino acids may enhance ghrelin action which in turn enhances insulin and glucagon secretions in lactating cows.     

MC1R c.310G>‐ and c.871G > A determine the coat color of Kumamoto sub‐breed of Japanese Brown cattle

Coat color is one of the important factors characterizing breeds for domestic animals. Melanocortin 1 receptor (MC1R) is a representative responsible gene for this phenotype. Two single‐nucleotide polymorphisms (SNPs) in bovine MC1R gene, c.296T > C and c.310G>‐, have been well characterized, but these SNPs are not enough to explain cattle coat color. As far as we know, MC1R genotypes of Kumamoto sub‐breed of Japanese Brown cattle have not been analyzed. In the current study, genotyping for c.296T > C and c.310G>‐ was performed to elucidate the role of MC1R in determining the coat color of this sub‐breed. As a result, most animals were e/e genotype, suggesting the coat color of this sub‐breed is derived from the e allele of MC1R gene. However, we found six animals with E/e genotype, which coat color would be black theoretically. Subsequently, sequence comparison was performed with these animals to identify other polymorphisms affecting coat color, elucidating that these animals possessed the A allele of c.871G > A commonly. c.871G > A was a non‐synonymous mutation in the seventh transmembrane domain, suggesting alteration of the function and/or the structure of MC1R protein. Our data indicated that the A allele of c.871G > A might be a loss‐of‐function mutation.     

Use of heat tolerance traits in discriminating between groups of sheep in central Brazil

The animal and its environment make up an integrated system, where each acts on the other. Tropical regions are characterized by high levels of solar radiation and environmental temperature which may adversely affect animal production. This study carries out a multivariate analysis of physical and physiological traits in sheep in the Federal District of Brazil to test the ability to separate groups of animals and determine which traits are most important in the adaptation of animal to heat stress. The variables studied included coat thickness, number and length of hairs, pigmentation of the skin and coat, number of sweat glands as well as heart and respiratory rates, rectal and skin temperatures, sweating rate, and blood parameters. Five groups of ten animals were used depending on breed (Bergamasca, crossbred, or Santa Inês) or coat color (Santa Inês—brown, white, and black). The data underwent multivariate statistical analyses including cluster, discriminate, and canonical, using Statistical Analysis System—SAS®. The tree diagram showed clear distances between groups studied and canonical analysis was able to separate individuals in groups, especially Bergamasca and white Santa Inês. The canonical correlation redundancy analysis showed that coat reflectance as well as hair length and number of hairs per unit area were the most useful in explaining changes in physiological traits. Skin and coat traits such as hair length, coat reflectance, percentage of epithelial area occupied by sweat glands, skin reflectance and thickness, as well as heart and breathing rates were the most important in separating these groups.