GnRHR gene polymorphisms and their effects on reproductive performance in Chinese goats

In this study, polymorphisms in the goat GnRHR gene exon 1 were detected by PCR-SSCP and DNA sequencing methods in 786 individuals from two different goat breeds. Two haplotypes (A and B), two observed genotypes (AA and AB), and two single nucleotide polymorphisms (SNPs) were detected, which resulted in five amino acid substitutions. The frequencies of haplotypes A and B in the two goat breeds were 0.78–0.83 and 0.17–0.22, respectively. The SNP locus was in Hardy–Weinberg disequilibrium in the two goat breeds (P < 0.05). Polymorphisms of the GnRHR gene were shown to be associated with litter size in the two goat breeds. The SNPs in the goat GnRHR gene had significant effects on litter size (P < 0.05). Therefore, these results suggest that the GnRHR gene is a strong candidate gene that affects litter size in goat.     

Sows’ responses to increased heat load – A review

There is a comprehensive body of literature on how increased air temperature affects the physiology, production and behaviour of sows, while very few studies consider the thermal effects of air humidity and air velocity.This review summarises studies that have investigated effects of air temperature by reviewing published literature in which sows were exposed to at least two different levels of air temperature ranging from 15 °C to 39 °C. Increased rectal temperature was investigated in the majority of the studies (26) and on average, the rectal temperature increased by 0.099 °C per °C increased air temperature above 25 °C. The increase was smaller at lower air temperatures, and it was suggested that rectal temperature is practically unaffected by air temperatures in the range of 15 °C–21 °C. This review elucidates how air temperature also affects performance indicators such as respiration rate, vaginal temperature, skin temperature, feed intake, milk yield, body weight loss during lactation, mortality, litter daily weight gain during lactation and sow behaviour.One study reported how respiration rate, rectal temperature, vaginal temperature and skin temperature were affected by both air temperature and air humidity, and the results suggest that the relative significance of air temperature and humidity may be similar for sows and finishing pigs (e.g. an increase of 40% relative humidity at an air temperature of 30 °C has a similar effect as a 1.9 °C increase in temperature).Studies on mitigation methods against the effects of high temperature and humidity such as snout cooling, drip cooling and floor cooling were reviewed to extract knowledge related to the effects of air velocity, temperatures of surrounding surfaces and the opportunity for sows to moisten their skin.     

Seasonal effect in expression of AQP1, AQP3 and AQP5 in skin of Murrah buffaloes

Aquaporins are transmembrane protein channels which are known to help the passage of water and solutes across the cell membranes. AQP1, AQP3 and AQP5 are isoforms of aquaporin known to aid in transepithelial water movement. AQP3 is also known to aid in glycerol transport. The present study was conducted to investigate the role of AQP1, AQP3 and AQP5 in thermoregulation of buffaloes by probing the expression of the genes in skin of buffaloes during different season viz. winter, spring and summer. The skin tissue samples were collected from the neck region of Murrah buffaloes (n = 12) and analyzed for gene expression by RT-PCR and immunolocalization. The physiological responses including respiration rate, rectal temperature and neck skin temperature observed during summer were significantly higher than winter and spring seasons. The study revealed the expression of AQP1, AQP3 and AQP5 genes in skin samples. The relative mRNA expressions of AQP1, AQP3 and AQP5 in skin relative to spring season were 1.41 ± 0.47, 1.95 ± 0.22 and 6.77 ± 1.02 folds during summer which were significantly higher than other seasons. The up-regulation of the expression of the studied AQPs were concomitant with the increase in physiological responses including skin temperature and sweating rate during summer. During summer season, AQP1 were mostly immunolocalized in the walls of skin blood capillaries, while AQP3 were observed mostly in the epidermal layer of the skin. The immunolocalization of AQP5 were mostly observed in the secretory glands of skin. The up-regulation of AQP1, AQP3 and AQP5 in skin during summer season indicates their role in thermoregulation of buffaloes.     

QTL and association analysis for skin and fibre pigmentation in sheep provides evidence of a major causative mutation and epistatic effects

The pursuits of white features and white fleeces free of pigmented fibre have been important selection objectives for many sheep breeds. The cause and inheritance of non‐white colour patterns in sheep has been studied since the early 19th century. Discovery of genetic causes, especially those which predispose pigmentation in white sheep, may lead to more accurate selection tools for improved apparel wool. This article describes an extended QTL study for 13 skin and fibre pigmentation traits in sheep. A total of 19 highly significant, 10 significant and seven suggestive QTL were identified in a QTL mapping experiment using an Awassi × Merino × Merino backcross sheep population. All QTL on chromosome 2 exceeded a LOD score of greater than 4 (range 4.4–30.1), giving very strong support for a major gene for pigmentation on this chromosome. Evidence of epistatic interactions was found for QTL for four traits on chromosomes 2 and 19. The ovine TYRP1 gene on OAR 2 was sequenced as a strong positional candidate gene. A highly significant association (P < 0.01) of grandparental haplotypes across nine segregating SNP/microsatellite markers including one non‐synonymous SNP with pigmentation traits could be shown. Up to 47% of the observed variation in pigmentation was accounted for by models using TYRP1 haplotypes and 83% for models with interactions between two QTL probabilities, offering scope for marker‐assisted selection for these traits.     

Effects of genetic variability of the dairy goat growth hormone releasing hormone receptor (<Emphasis Type="Italic">GHRHR</Emphasis>) gene on growth traits

Growth hormone-releasing hormone receptor (GHRHR) plays a critical role in growth hormone (GH) synthesis, release and regulation of pituitary somatotroph expansion in vertebrates. The objective of this study was to investigate variations in goat GHRHR gene and their associations with growth traits in 668 dairy goats. The results showed four novel single nucleotide polymorphisms (SNPs): NC_007302:g.5203C>T, 7307C>G, 9583G>A and 9668A>C. In detail, the novel SNP C>T in the 5203rd nucleotide identified a missense mutation: CCC (Pro)>TCC (Phe) at position 116aa of the goat GHRHR (423aa). Besides, 9583G>A and 9668A>C polymorphism were in complete linkage disequilibrium. The genetic diversity analysis revealed that the Guanzhong dairy goat possessed intermediate genetic diversity in P3 and P7 loci, and the Xinong Sannen dairy goat belonged to poor genetic diversity in P4 locus. Significant associations between the genotypes of P3 locus and body length, body height and chest circumference was observed in Guanzhong goat (P < 0.05). However, in Xinong saanen population, significant statistical difference was only found in body height and body length (P < 0.05). In P4 and P7 loci, no significant associations were detected between any variant sites and body length, body height and chest circumference, as well as for the milk traits (P > 0.05). These results strongly suggested that the goat GHRHR gene is a candidate gene that influences growth traits in dairy goat.     

Effects of caffeine on blood pressure, heart rate, and forearm blood flow during dynamic leg exercise

This study examined the acute effects of caffeine on thecardiovascular system during dynamic leg exercise. Ten trained,caffeine-naive cyclists (7 women and 3 men) were studied at rest andduring bicycle ergometry before and after the ingestion of 6 mg/kgcaffeine or 6 mg/kg fructose (placebo) with 250 ml of water. Afterconsumption of caffeine or placebo, subjects either rested for 100 min(rest protocol) or rested for 45 min followed by 55 min of cycleergometry at 65% of maximal oxygen consumption (exercise protocol).Measurement of mean arterial pressure (MAP), forearm blood flow (FBF),heart rate, skin temperature, and rectal temperature and calculation offorearm vascular conductance (FVC) were made at baseline and at 20-minintervals. Plasma ANG II was measured at baseline and at 60 minpostingestion in the two exercise protocols. Before exercise, caffeineincreased both systolic blood pressure (17%) and MAP (11%) withoutaffecting FBF or FVC. During dynamic exercise, caffeine attenuated theincrease in FBF (53%) and FVC (50%) and accentuated exercise-inducedincreases in ANG II (44%). Systolic blood pressure and MAP were alsohigher during exercise plus caffeine; however, these increases weresecondary to the effects of caffeine on resting bloodpressure. No significant differences were observed inheart rate, skin temperature, or rectal temperature. These findingsindicate that caffeine can alter the cardiovascular response to dynamicexercise in a manner that may modify regional blood flow andconductance.

     

Novel SNPs in the <Emphasis Type="Italic">ATP1B2</Emphasis> gene and their associations with milk yield,milk composition and heat-resistance traits in Chinese Holstein cows

Genetic association analysis was applied to examine the effect of the Na⁺/K⁺-ATPase beta 2 subunit (ATP1B2) gene on rectal temperature, milk traits, K⁺ levels and Na⁺/K⁺-ATPase (NKA) activity in the red blood cells of 1001 Chinese Holstein cows under normal and heat-stress conditions. We detected two novel single nucleotide polymorphisms, G2258A and C2833T, in the second and fourth introns, respectively, of ATP1B2. G2258A significantly affected milk fat content (P < 0.05) and 305-day milk yield (P < 0.01), but not milk protein content. C2833T significantly affected milk protein content (P < 0.01) and 305-day milk yield (P < 0.05), but not milk fat content. Calculated gene substitution effects suggested that A to G substitution in G2258A, and T to C substitution in C2833T, positively affected milk fat content, 305-day milk yield and somatic cell score, but negatively affected milk protein content. We also detected significant variation in milk fat content, milk protein content, 305-day milk yield and somatic cell scores (P < 0.05 or P < 0.01) among the nine ATP1B2 haplotypes. Under heat-stress, the C2833T polymorphism was significantly related to rectal temperature (P < 0.01), red blood cell K⁺ levels, NKA activity and milk yield (P < 0.05). Cows with the TT genotype showed the desirable characteristics of low rectal temperature and red blood cell K⁺, low decline rate in milk yield and red blood cell NKA activity. This study suggests that the ATP1B2 single nucleotide polymorphism C2833T is a genetic marker of heat-resistance traits in Chinese Holstein cows.     

Identification of novel SNPs in the Sarda breed goats POU1F1 gene and their association with milk productive performance

The aim of the study was to detect polymorphism in the POU1F1 gene in Sarda breed goat, as well as to establish if SNPs could be associated with milk productive traits. The research was conducted on 129 Sarda breed goats from 4 to 5 years old, multiparous, lactating and in their third to fifth lactation. We report nine exonic and seven non-coding regions SNPs within the Sarda goat POU1F1 gene, namely, Ex 1 61 G>C; Ex 1 108 G>A; Ex 3 C>T; Ex 3 92 C>T; Ex 4 110 A>G; Ex 5 34 G>A resulting in Arg213Lys change; IVS4 641 G>A, IVS4 643 A>C, IVS4 659 G>A, IVS4 677 A>C, IVS4 G699Del, IVS4 709 C>G, Ex 6 17 G>A resulting in Arg228Ser change, Ex 6 58 G>T, Ex 6 172 T>C, 3′UTR 110 T>C. A statistically significant association was found between genotype TT, in position 17 of the exon 6 (3.1 % of frequency), and increased milk yield (P < 0.01) while genotype GT (25.6 % of frequency) was associated with a higher fat content. Genotype TT in position 58 of the exon 6 (3.9 % of frequency) was found to be associated with a higher fat (P < 0.01) and protein content (P < 0.05). Twenty-eight haplotypes were detected, but no significant association between the haplotypes and the milk production traits have been found. Our data, as well as providing new SNPs extending the POU1F1 gene characterization, evidence a relationship between polymorphism and milk production traits in Sarda goat breed.     

Polymorphisms of caprine <Emphasis Type="Italic">POU1F1</Emphasis> gene and their association with litter size in Jining Grey goats

Seven pairs of primers were designed to amplify 5′ promoter region, six exons and partial introns and to detect the polymorphisms of POU1F1 gene in five goat breeds with different prolificacy. The results showed that six mutations were identified in caprine POU1F1 gene including C256T in exon 3, C53T and T123G in intron 3, and G682T (A228S), T723G and C837T in exon 6. The former four mutations were novel SNPs in goat POU1F1 gene. The 53 and 123 loci were in complete linkage disequilibrium in five caprine breeds. Regarding the 256 locus, the Jining Grey goat does with genotype CT had 0.66 kids more than those with genotype CC (P < 0.05), while does with genotype GT had 0.63 (P < 0.05) kids more than those with genotype GG at the 682 locus. The present study preliminarily showed an association between allele T at the 256 and 682 loci of POU1F1 gene and high litter size in Jining Grey goats. Totally 16 haplotypes and 50 genotypes were identified at the above six loci in POU1F1 gene of five goat breeds. Three common haplotypes (hap2, hap3 and hap4) were identified in five goat breeds joined. Four specific haplotypes (hap7, hap9, hap11 and hap13) were detected in Jining Grey goats. The predominant haplotype was hap1 (35.29% and 48.25%) in both Jining Grey and Guizhou White goats, while hap4 (50%) in Boer goats, and hap2 (42.86% and 38.75%) in both Wendeng Dairy and Liaoning Cashmere goats. The most frequent genotypes at six loci in the above five goat breeds were hap1/hap2 (14.38%) and hap1/hap4 (14.38%), hap1/hap2 (38.60%), hap4/hap4 (40.91%), hap2/hap4 (26.53%), hap2/hap5 (20.00%), respectively. The Jining Grey goat does with nine genotypes analyzed of POU1F1 gene showed no obvious difference in litter size.     

Regional differences in sweat rate response of steers to short-term heat stress

Six Angus steers (319 ± 8.5 kg) were assigned to one of two groups (hot or cold exposure) of three steers each, and placed into two environmental chambers initially maintained at 16.5–18.8°C air temperature (T _a). Cold chamber T _a was lowered to 8.4°C, while T _a within the hot chamber was increased to 32.7°C over a 24-h time period. Measurements included respiration rate, and air and body (rectal and skin) temperatures. Skin temperature was measured at shoulder and rump locations, with determination of sweat rate using a calibrated moisture sensor. Rectal temperature did not change in cold or hot chambers. However, respiration rate nearly doubled in the heat (P < 0.05), increasing when T _a was above 24°C. Skin temperatures at the two locations were highly correlated (P < 0.05) with each other and with T _a. In contrast, sweat rate showed differences at rump and shoulder sites. Sweat rate of the rump exhibited only a small increase with T _a. However, sweat rate at the shoulder increased more than four-fold with increasing T _a. Increased sweat rate in this region is supported by an earlier report of a higher density of sweat glands in the shoulder compared to rump regions. Sweat rate was correlated with several thermal measurements to determine the best predictor. Fourth-order polynomial expressions of short-term rectal and skin temperature responses to hot and cold exposures produced r values of 0.60, 0.84, and 0.98, respectively. These results suggest that thermal inputs other than just rectal or skin temperature drive the sweat response in cattle.     

Characterisation of the genetic effects of the ADFP gene and its association with production traits in dairy goats

Adipose differentiation-related protein (ADFP) is important for regulation of lipid metabolism and insulin secretion in beta-cells. In this study, we investigated polymorphisms within the caprine ADFP gene and determined its relationship with production traits. As there was no sequence information available for the caprine ADFP gene, we generated DNA sequence data and examined the genomic organisation. The caprine ADFP gene is organised into 7 exons and 6 introns that span approximately 8.7 kbp and is transcribed into mRNA containing 1353 bp of sequence coding for a protein of 450 amino acids. The protein sequences showed substantial similarity (71–99%) to orthologues from cattle, human and mouse. We identified polymorphisms in the sequences using DNA sequencing, PCR-RFLP and forced PCR-RFLP methods. Seven single nucleotide polymorphisms (SNPs) were identified using samples from 4 different goat populations consisting of 1408 healthy and unrelated individuals. Six haplotypes involving the 7 SNPs from the caprine ADFP gene were identified and their effects on production traits were analysed. Haplotype 6 had the highest haplotype frequency and was highly significantly associated with chest circumference and milk yield in the analysed populations. The results of this study suggest that the ADFP gene is a strong candidate gene affecting production traits and may be used for marker-assisted selection and management in Chinese dairy goat breeding programmes.     

Heat stress of gilts around farrowing causes oxygen insufficiency in the umbilical cord and reduces piglet survival

Late gestating sows are susceptible to high ambient temperatures, possibly causing farrowing complications and reducing piglet survival. This experiment aimed to quantify in the days leading up to farrowing the impact of sow heat stress (HS) on farrowing physiology and survival of the piglets. Pregnant primiparous sows (gilts) were allocated to either thermoneutral control (CON, n = 8; constant 20 °C) or cyclical HS conditions (n = 8; 0900 h to 1700 h, 30 °C; 1700 h to 0900 h, 28 °C) from d 110 of gestation until farrowing completion. Gilt respiration rate, skin temperature and rectal temperature were recorded daily, and farrowing duration was quantified by video analyses. Blood samples were collected from the piglet umbilical vein at birth. At 48 h of age, piglet growth was quantified by morphometric analyses. The thermal exposure model induced HS and respiratory alkalosis in the gilts, as indicated by increased respiration rate, rectal temperature, skin temperature (all P < 0.001), plasma cortisol (P = 0.01) and blood pH (P < 0.001). Heat-stressed gilts took longer to start expelling placentae (P = 0.003), although the active farrowing duration was not significantly different between treatments. Stillbirth rates were higher in the HS group (P < 0.001), with surviving piglets at birth having lower umbilical vein partial pressure of oxygen (P = 0.04), oxygen saturation rate (P = 0.03) and tending to have increased lactate concentrations (P = 0.07). At birth, piglet skin meconium staining scores were greater in the HS group (P = 0.022). At 48 h of age, piglets from the HS group had reduced small intestinal length (P = 0.02), reduced jejunal crypt depth (P = 0.02) and lighter absolute brain weight (P = 0.001). In contrast, piglet BW, growth rate, relative organ weight and small intestinal mucosal barrier function did not change between treatments. Collectively, these findings demonstrated gilt HS during late gestation caused farrowing complications and reduced the umbilical oxygen supply to the piglets at parturition, leading to increased risks of piglet stillbirth with implications on impaired neonatal survivability and development.     

A directed search around caprine candidate loci provided evidence for microsatellites linkage to growth and cashmere yield in Rayini goats

Four genomic regions located on goat chromosomes 1, 2, 5 and 13 previously reported as significant DNA regions harboring quantitative trait loci (QTL) for growth and fleece traits in goat were considered as candidate genomic regions. The objective of conducting this study was to determine whether there is evidence for association of growth traits and cashmere yield in native Iranian Rayini goats with markers flanking these candidate regions. One hundred and twenty kids from 6 Rayini bucks (20 half-sib offspring per buck) were used in the present study. The six bucks and their offspring were genotyped for eight microsatellite markers flanking the candidate intervals. Body weights recorded on the kids were birth weight (BW0), weaning weight (BW3) and weight at 6 months of age (BW6). Average daily gains from birth to weaning and weaning to 6 months of age were computed based on the values obtained for BW0, BW3 and BW6. Cashmere yield was also recorded for the kids at the first, second and third year of age. QTL analysis in the candidate intervals was conducted using the least squared regression interval mapping approach. Linkage analysis indicated significant QTL for growth traits on goat chromosomes 1, 2 and 5 (chromosome-wide significance of P < 0.01). Three of the candidate regions located on chromosomes 2, 5 and 13 were linked to cashmere yield. QTL effect ranged from 0.8 to 4.3 in units of residual standard deviation for different traits and families.     

Individual variation in body temperature and energy expenditure in response to mild cold

We studied interindividual variation in body temperature and energy expenditure, the relation between these two, and the effect of mild decrease in environmental temperature (16 vs. 22 degrees C) on both body temperature and energy expenditure. Nine males stayed three times for 60 h (2000-0800) in a respiration chamber, once at 22 degrees C and twice at 16 degrees C, in random order. Twenty-four-hour energy expenditure, thermic effect of food, sleeping metabolic rate, activity-induced energy expenditure, and rectal and skin temperatures were measured. A rank correlation test with data of 6 test days showed significant interindividual variation in both rectal and skin temperatures and energy expenditures adjusted for body composition. Short-term exposure of the subjects to 16 degrees C caused a significant decrease in body temperature (both skin and core), an increase in temperature gradients, and an increase in energy expenditure. The change in body temperature gradients was negatively related to changes in energy expenditure. This shows that interindividual differences exist with respect to the relative contribution of metabolic and insulative adaptations to cold.     

Association between cholecystokinin type A receptor haplotypes and growth traits in Japanese Hinai-dori crossbred chickens

We previously identified quantitative trait loci for body weight and average daily gain in a common region between MCW0240 (chr 4: 69.9 Mb) and ABR0622 (chr 4: 86.3 Mb) on chicken chromosome 4 in an F₂ resource population produced by crossing low- and high-growth lines of the Hinai-dori breed. Cholecystokinin type A receptor (CCKAR) is a candidate gene affecting growth traits in the region. In this study, we genotyped polymorphisms of the CCKAR gene and investigated its association with growth traits in a Hinai-dori F₂ intercross population. All the exons of the CCKAR gene in the parental population were subjected to PCR amplification, nucleotide sequenced and haplotypes identified. To distinguish resultant diplotype individuals in the F₂ population, a mismatch amplification mutation assay was performed. Five haplotypes (Haplotypes 1–5) were accordingly identified. Six genotypes produced by the combination of three haplotypes (Haplotype 1, 3, and 4) were examined in order to identify associations between CCKAR haplotypes and growth traits. The data indicate that Haplotype 1 was superior to Haplotype 3 and 4 in body weight at 10 and 14 weeks of age, average daily gain between 4 and 10 weeks, 10 and 14 weeks, and 0 and 14 weeks of age. It was concluded that CCKAR is a useful marker of growth traits and could be used to develop strategies for improving growth traits in the Hinai-dori breed.     

A gating mechanism of the BsYetJ calcium channel revealed in an endoplasmic reticulum lipid environment

The transmembrane BAX inhibitor-1-containing motif 6 (TMBIM6) is suggested to modulate apoptosis by regulating calcium homeostasis in the endoplasmic reticulum (ER). However, the precise molecular mechanism underlying this calcium regulation remains poorly understood. To shed light on this issue, we investigated all negatively charged residues in BsYetJ, a bacterial homolog of TMBIM6, using mutagenesis and fluorescence-based functional assays. We reconstituted BsYetJ in membrane vesicles with a lipid composition similar to that of the ER. Our results show that the charged residues E49 and R205 work together as a major gate, regulating calcium conductance in these ER-like lipid vesicles. However, these residues become largely inactive when reconstituted in other lipid environments. In addition, we found that D195 acts as a minor filter compared to the E49-R205 dyad. Our study uncovers a previously unknown function of BsYetJ/TMBIM6 in the calcium-dependent inactivation of BsYetJ, providing a framework for the development of a lipid-dependent mechanistic model of BsYetJ that will facilitate our understanding of calcium-dependent apoptosis.     

Polymorphism in <Emphasis Type="Italic">BMP4</Emphasis> gene and its association with growth traits in goats

Bone morphogenetic protein 4 (BMP4) plays a crucial role in growth and development of mammals, especially for bone growth. The aim of the present study was to assess the association of polymorphism in BMP4 gene with growth traits in three goat breeds. In this study, on the basis of PCR-SSCP and DNA sequencing methods, two new silent mutation sites of A1986G and G2203A were identified in the intron 2, and one novel microsatellite of dinucleotide-repeated (CA) sequence was in the 3′ flanking region. Associations between growth traits and BMP4 gene polymorphism were investigated, and significant statistical association results were found in body height, chest circumference and trunk index (P < 0.05). The result strongly suggested that the microsatellite located in BMP4 gene might be in linkage with the two new SNPs. Further investigations are required for detecting the polymorphism of this gene in a broad variety of goat breeds and populations Xingtang Fang and Haixia Xu contributed equally to this work.