PCR-SSCP analysis of leptin gene and its association with milk production traits in river buffalo (Bubalus bubalis)

Leptin gene has been found to be associated with various economic traits including milk production and fat quality in dairy animals. In the present study, we investigated genetic variations in intron 1 region of leptin gene in riverine buffaloes (Bubalus bubalis) using polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP) and sequencing methods and associated them with milk traits. The study revealed three SSCP variants A, B and C among a total of 301 buffaloes from nine breeds. The frequency of variant C was found invariably high among all the breeds except in Marathwada buffalo. Variant A was found to be absent in Chilika, Nili-Ravi, Nagpuri and Pandharpuri breeds and also had the lowest frequencies in Mehsana, Jaffarabadi, Murrah and Toda breeds. Sequencing of SSCP variants revealed a total of five polymorphic sites, with three haplotypes. Statistical analysis revealed significantly high fat percentage at 150?days in SSCP variant B in Mehsana buffaloes. However, the associations of SSCP variants of leptin gene with total milk yield, 305?days milk yield and total fat yield were found to be non-significant. The present study is the first report on association analysis of leptin gene polymorphisms with milk production and milk quality traits in river buffalo.     

Population structure and phylogeography of Toda buffalo in Nilgiris throw light on possible origin of aboriginal Toda tribe of South India

We report the genetic structure and evolutionary relationship of the endangered Toda buffalo of Nilgiris in South India with Kanarese and two other riverine buffalo breeds. The upgma phylogeny drawn using Nei’s distance grouped South Kanara and Toda buffaloes at a single node while Marathwada and Murrah together formed a separate node. Principal component analysis was performed with pairwise interindividual chord distances which revealed clustering of Murrah and Marathwada buffaloes distinctly, while individuals of Toda and South Kanara breeds completely intermingled with each other. Furthermore, there were highly significant group variances (p < 0.01) when the breeds were grouped based on phylogeny, thus revealing the existence of cryptic genetic structure within these buffalo breeds. To know the evolutionary relationship among these breeds, 537‐bp D‐loop region of mitochondrial DNA was analysed. The phylogenetic analysis of mtDNA haplotypes following NJ algorithm with Chinese swamp buffalo as outgroup revealed a major cluster that included haplotypes from all the four investigated breeds and two minor clusters formed by South Kanara and Toda haplotypes. Reduced median network analysis revealed haplotypes of South Kanara and Toda to be quite distinct from the commonly found haplotypes indicating that these might have been ancestral to all the present‐day haplotypes. Few mutations in two of the haplotypes of South Kanara buffalo were found to have contributed to ancestral haplotypes of Toda buffalo suggesting the possible migration of buffaloes from Kanarese region towards Nilgiris along the Western Ghats. Considering the close social, economic and cultural association of Todas with their buffaloes, the present study supports the theory of migration of Toda tribe from Kanarese/Mysore region along with their buffaloes.     

广西的尼里—拉菲水牛和摩拉水牛mtDNA D-loop区遗传多样性研究

[目的]探究引入广西南宁的尼里-拉菲水牛和摩拉水牛的mtDNA D-loop区遗传多样性与母系起源。[方法]采用PCR扩增、测序及生物信息学方法。[结果]对从广西南宁采集的52个江河型水牛（尼里-拉菲水牛25个,摩拉水牛27个）mtDNA D-loop序列与GenBank下载的20条尼里-拉菲水牛和23条摩拉水牛序列进行联合分析,共检测到112个变异位点,定义42个单倍型,发现摩拉水牛（Hd: 0.934±0.027）与尼里-拉菲水牛的遗传多样性（Hd: 0.929±0.017）都很丰富。NJ系统发育树显示尼里-拉菲水牛和摩拉水牛含有江河型和沼泽型水牛mtDNA支系,表明尼里-拉菲水牛和摩拉水牛引入中国后均与沼泽型水牛进行了杂交,在外貌上很难区分。[结论]引入广西南宁的尼里-拉菲水牛和摩拉水牛与沼泽型水牛存在广泛的血缘混杂现象。     

Genetic and morphometric characterization of a local Vietnamese Swamp Buffalo population

The water buffalo plays a key role in the socio-economy of South-East Asia as it is the main draught power for paddy rice cultivation. While in the Indian subcontinent the water buffalo is the riverine type, in South-East Asia the majority of buffaloes are of the swamp type. In the poor remote northern province of Ha Giang in Vietnam, improvement of the swamp buffalo breed may be one of the best ways to increase sustainability of farming systems. Therefore, analysis of the genetic structure of the province buffalo population is a prerequisite to any conservation or improvement project. A total of 1122 animals were described for 11 body and horn measurements for morphometric characterization. From this sample set, 744 animals were genotyped for 17 microsatellite markers. Also 17 animals from southern provinces of Vietnam were genotyped as a comparative sample. The results showed that genetic diversity as well as inbreeding value in the Ha Giang was high. The F _ST values within the province and across Vietnam were low indicating that most of the population variation is explained by individual variability. Bayesian clustering analysis did not highlight the presence of subdivided populations. These results are useful for the implementation of a conservation and improvement strategy of the swamp buffalo in order to guarantee the householders' needs for sustainability of the farming system in the Ha Giang province.     

Evaluation of direct and indirect measures of quarter milk from crossbred buffaloes

Somatic cell count (SCC), bacteriological findings and electrical conductivity (EC) were measured in milk from 216 quarters of 54 Murrah crossbred buffaloes for characterization of buffalo milk. Mean SCC in normal, subclinical and clinical mastitic milk was 171, 799 and 6039 × 10³/mL, respectively. Coagulase negative Staphylococci (CNS) were the major organisms associated with subclinical mastitis, whereas CNS and coliforms were the predominant organisms in clinical mastitis. EC values were found to be significantly higher in buffalo milk with mammary infection. The best trade-off between sensitivity and specificity for diagnosing mastitis in buffaloes by EC score was found at the cut-off value of 3.7 mS/cm. The direct measures such as SCC, EC, bacteriological findings, and appearance of milk were found to be useful for the evaluation of udder health of buffaloes and categorization of milk.     

Pregnancy-associated Glycoprotein Profile during the First Trimester of Pregnancy in Egyptian Buffalo Cows

Pregnancy-associated glycoprotein (PAG) concentrations were measured in buffalo cows starting from day 28 after breeding. Oestrus was synchronized in 10 buffaloes using two injections of 25 mg prostraglandin (PG)F_{2 α} (Lutalyse^®) at a 11-day interval. Blood sampling was conducted nearly twice weekly. Results indicated that plasma PAG concentrations in non-pregnant buffaloes were low (<0.20 ng/ml) during the whole experimental period (day 28 to 103), while in pregnant animals plasma PAG levels increased from day 28 (4.48 ± 0.92 ng/ml) until day 41 (27.27 ± 6.74 ng/ml), remaining high (20.71 ± 9.20 ng/ml) until day 103. Progesterone levels were significantly (p < 0.0001) higher in pregnant (3.51–4.80 ng/ml) than in non-pregnant buffaloes (0.28–1.52 ng/ml). A significant difference (p < 0.0001) in plasma PAG concentrations between pregnant and non-pregnant animals starting at day 28 after breeding suggests that PAG-radioimmunoassay could be suitable for pregnancy diagnosis in buffaloes during this period. In conclusion, PAG test offers the advantages that it requires a single plasma sample for early pregnancy diagnosis as well as the accuracy of the test for the detection of pregnancy as early as day 28.     

Full-term delivery of river buffalo calves (2n = 50) from in vitro-derived vitrified embryos by swamp buffalo recipients (2n = 48)

Field trials were carried out on the use of swamp buffaloes (2n = 48) as recipients of in vitro-derived vitrified river buffalo (2n = 50) embryos. River buffalo embryos were produced through a total in vitro production system using frozen semen from a top progeny-tested bull and slaughter-house-derived ovaries and cryopreserved by a vitrification technique. Recipient swamp buffaloes were Philippine carabaos selected on body condition, parity, and reproductive status and owned by village farmers. Eighty post-warmed riverine embryos were transferred non-surgically to 40 swamp buffalo recipients, of which three healthy riverine calves were born by normal delivery and one stillbirth, representing a 5.0% (4/80) full-term development and 10% (4/40) calving rate with 75% (3/4) normal and 25% (1/4) stillbirth. The results demonstrate that, under field conditions and despite the difference in chromosome number between the embryo and recipient, swamp buffaloes are potential surrogate mothers of imported, genetically superior river buffalo frozen embryos. Thus, embryo IVP-vitrification and transfer technology is a potential tool in the propagation of genetically superior buffaloes. Precautionary measures, however, are recommended to avoid dystocia and stillbirth.     

Transcript profiling of pattern recognition receptors in a semi domesticated breed of buffalo, Toda, of India

The primary objective of this study was to assess the expression profile and levels of toll-like receptor (TLR) mRNAs in the spleen, lung, mediastinal lymph node (MLN), jejunum, rectum, skin and peripheral blood mononuclear cells (PBMC) of Toda and Murrah buffalos. Spleen and PBMC had increased expression of TLR mRNAs 2, 4, 5, 6, 8, 9 and 10; lung had increased expression of TLR mRNAs 2, 4, 5, 6 and 8, MLN TLR mRNA 6, 9, 10 and decrease in TLR 3 and 7 mRNAs in skin. No significant differences were observed in the expression levels of any of the TLR mRNA in jejunum and rectum. Toda buffaloes showed significantly higher expression levels of TLR 9 mRNA in MLN, TLR mRNAs 1, 5, 6, 9 and 10 in skin and TLR mRNAs 2, 4, 7 and 9 in PBMC than Murrah buffaloes living in the vicinity. Toda and Murrah buffaloes were inoculated with TLR5 (flagellin) and TLR9 (CpG ODN) ligands in vivo and expression levels of the respective TLRs analyzed 12h later. Following CpG inoculation, Toda buffaloes had significantly higher levels of TLR 9 mRNA expression but not in Murrah. However, flagellin induction did not increase TLR 5 mRNA expression in both these breeds. Histological sections of the skin were made and infiltrating cell clusters were graded and quantified. Following CpG inoculation, Toda buffaloes showed higher numbers of infiltrating grade 1 and grade 3 cell clusters while Murrah showed lower numbers of infiltrating grade 1 cells as compared to mock-inoculated skin sections. Flagellin treatment revealed no significant differences in infiltrating cell clusters in both the breeds. The results have shown differential expression of TLR mRNAs in various tissues between two divergent buffalo breeds with the highest difference in TLR expression profile seen in the skin, the largest portal of entry of pathogens, of Toda.     

Evaluation of genetic variability and mutation drift equilibrium of Banni buffalo using multi locus microsatellite markers

The present study was conducted to evaluate genetic diversity of Banni buffalo and its relationship/differentiation with Murrah using genotypic data on 24 heterologus bovine specific microsatellite marker loci. A total of 138 alleles were observed with a mean of 5.75 alleles/locus across two populations. The mean observed and expected heterozygosities were found to be 0.441 and 0.572 respectively in Banni buffaloes while it was 0.464 and 0.610 respectively in Murrah buffaloes. The average heterozygosity deficit was significantly positive with substantially higher values observed in Banni (22.3%) and Murrah (24%) buffalo populations. Banni buffalo population, when evaluated for mutation drift equilibrium revealed significant heterozygosity excess under IAM while no such excess was observed under SMM and TPM. The qualitative graphical test revealed a normal L-shaped distribution of allele frequencies indicating the absence of genetic bottleneck in Banni buffaloes. The mean estimates of F-statistics over all the loci were 0.376 for F_IT, 0.187 for F_ST and 0.232 for F_IS respectively. Analysis of molecular variance (AMOVA) revealed 18.95% of the total variation being explained by between breed differences while 14.36% of the variation explained differences between individuals within each breed. Genotype assignment test revealed distinct clustering of Banni and Murrah buffaloes. Genetic distance was estimated using three different methods, the results of which revealed considerable genetic differentiation between these two buffalo populations. The divergence time between Banni and Murrah buffaloes was estimated to be around 7286 years. The results of the present study may be helpful in decision making for conservation programs as Banni buffalo population is on decline.     

摩拉水牛及德宏水牛瘤胃产甲烷菌多样性比较

本试验旨在比较分析摩拉水牛及德宏水牛瘤胃液中产甲烷菌的多样性。选取健康雌性摩拉水牛及德宏水牛各3头,采用口腔导管法收集瘤胃液,酚-氯仿-异戊醇抽提法提取瘤胃液总DNA,用产甲烷菌特异性引物Met86F/Met1340R扩增产甲烷菌16S rDNA,构建16S rDNA基因克隆文库。摩拉水牛及德宏水牛各获得96个16S rDNA基因序列,均归类于Methanobacteriales目,其中德宏水牛有82个序列（18个OTUs）与已知菌的16S rDNA序列相似性≥ 97%,占总序列的85.4%,有14个序列（9个OTUs）与已知菌16S rDNA序列相似性为89%~97%;摩拉水牛有94个序列（13个OTUs）与已知菌16S rDNA序列相似性≥ 97%,占总序列的97.9%,仅有2个序列（1个OTUs）与已知菌16S rDNA序列相似性为94%。系统进化树分析表明,所有序列分别聚集于两大分支上,其中德宏水牛有13个OTUs代表序列和摩拉水牛7个OTUs代表序列聚集在进化树顶端的同一分支上,且在系统发育距离上与Methanobacteriales目中任何已知相似序列都相隔较远。德宏水牛瘤胃中的SGMT簇序列和RO簇序列所占总序列比列分别为83.3%、9.4%,摩拉水牛瘤胃中的SGMT簇序列和RO簇序列所占总序列的比列分别为51.0%、9.4%。由此可见,摩拉水牛及德宏水牛瘤胃产甲烷菌序列以Methanobacteriales目为主,其中德宏水牛拥有更多未知的产甲烷菌序列;德宏水牛瘤胃中的SGMT簇产甲烷菌序列比例要高于摩拉水牛。     

摩拉水牛和尼里-拉菲水牛PRKAA2基因SNPs检测及遗传多样性分析

为研究水牛蛋白激酶AMP活化的催化亚基α2（protein kinase AMP-activated catalytic subunit alpha 2,PRKAA2）基因多态性,本试验以摩拉水牛和尼里-拉菲水牛基因组DNA为模板,扩增PRKAA2基因外显子4及内含子3部分序列,通过常规测序法检测其SNP并进行遗传多样性分析。结果发现,PRKAA2基因外显子4内存在1个SNP位点（c.462 G>A）,PRKAA2基因内含子3部分序列存在3个SNPs位点（IVS3.557 T>C、IVS3.560 C>T和IVS3.565 G>A）。经遗传多样性分析表明,在c.462 G>A位点的野生纯合型和杂合型比突变纯合型更有优势,IVS3.557 T>C和IVS3.560 C>T位点的突变纯合型为非优势基因型,IVS3.565 G>A位点杂合型为优势基因型。IVS3.565 G>A位点在摩拉水牛群体中处于Hardy-Weinberg非平衡状态;c.462 G>A位点在尼里-拉菲水牛群体中处于Hardy-Weinberg非平衡状态。4个SNPs位点在摩拉水牛群体中均为中度多态;c.462 G>A、IVS3.557 T>C位点在尼里-拉菲水牛群体中为低度多态,IVS3.560 C>T、IVS3.565 G>A位点为中度多态。IVS3.557 T>C位点在两个水牛群体中杂合度较低。说明摩拉水牛IVS3.565 G>A位点和尼里-拉菲水牛c.462 G>A位点的基因型频率和基因频率遗传状态不平衡,尼里-拉菲水牛群体中IVS3.557 T>C位点遗传变异小,选择潜力不高。4个多态位点可以构建5种单倍型,其中T-C-G-G是摩拉水牛群体和尼里-拉菲水牛群体的优势单倍型。综上,本研究检测的摩拉水牛和尼里-拉菲水牛PRKAA2基因上4个SNPs位点可为水牛标记辅助选择育种提供参考。     

In vitro kinetics of hepatic albendazole sulfoxidation in channel catfish (Ictalurus punctatus), tilapia (Oreochromis sp.), rainbow trout (Oncorhynchus mykiss) and induction of EROD activity in ABZ-dosed channel catfish

Liver microsomes from market-size ( n  = 6) rainbow trout, channel catfish and tilapia were used to investigate in vitro biotransformation kinetics of albendazole (ABZ). ABZ was transformed to a single metabolite, ABZ sulfoxide (ABZ-SO). Catfish displayed the highest maximal velocity ( V _max = 264.0 ± 58.6 pmols ABZ-SO/min/mg protein) followed by tilapia (112.3 ± 8.2) and rainbow trout (73.3 ± 10.3). V _max in catfish was significantly different ( P  < 0.05) from the other two species. Michaelis–Menten constant ( K _m) values (μ m ) varied significantly among the species: rainbow trout (3.9 ± 0.5), tilapia (9.2 ± 1.7) and catfish (22.0 ± 3.2). However, V _max/ K _m ratios showed no difference among the three species, making them equally efficient performing this phase I biotransformation reaction. In a second series of experiments, channel catfish ( n  = 6 per treatment) were dosed in vivo with gel-food containing ABZ (10 mg/kg, p.o.). Fish were killed at 24, 48, 72 and 120 h after dosage. Control fish were fed ABZ-free feed. Induction of ethoxyresorufin-o-deethylase activity was significant ( P  < 0.05) in all ABZ-dosed treatments as compared with controls.     

Microsatellite analysis of genetic diversity in the Catalonian donkey breed

A total of 111 individuals (79 females and 32 males) of the endangered Catalonian donkey breed were analysed by using a commercial equine paternity polymerase chain reaction typing kit. Eleven of the 12 horse microsatellites were amplified when using donkey's DNA. One locus, ASB2, did not amplify in any sample. The allele range for HTG4 overlapped with HMS7 and their results could not be interpreted. The mean number of alleles detected per locus was 7.7 (±1.0), being the average of genetic diversity detected in the population, and measured as the unbiased average expected heterozygosity ( H _e), of 0.712 ± 0.038. All the analysed loci showed disagreement with Hardy–Weinberg proportions except HTG7 (from permutation tests). The within-population-inbreeding estimate was highly significant (p < 0.001) and equal to 15.4% (as measured by F _IS-statistic). The cumulative exclusion probability was 0.999. The deficit of heterozygotes can be partly explained by a population subdivision effect ( F _ST=3.5%), although the main factor that has provoked this lack of heterozygotes can be attributed to consanguinity.     

Pharmacokinetics of indomethacin in sheep after intravenous and intramuscular administration

The pharmacokinetics of indomethacin (1mg/kg) was determined in six adult sheep after intravenous (i.v.) and intramuscular (i.m.) injection. Plasma concentrations were maintained within the therapeutic range (0.3–3.0 μg/mL) from 5 to 50 min after i.v. and from 5 to 60–90 min after i.m. administration. After two trials, indomethacin best fitted an open two-compartment model. The mean (±SD) volumes of distribution at steady state ( V d_ss) were 4.10 ± 1.40 and 4.21 ± 1.93 L/kg and the mean clearance values ( C l_B) were 0.17 ± 0.06 and 0.22 ± 0.12 L/h.kg for i.v. and i.m. routes, respectively. The elimination phase half-lives did not show any significant difference between routes of injection ( t _½β = 17.4 ± 4.6 and 21.25 ± 4.44 h, i.v. and i.m. respectively). After i.m. administration, plasma maximum concentration ( C _max =  1.10 ± 0.68 μg/mL) was reached 10 min after dosing; the absorption phase was fast ( K _ab = 26 ± 18 h-1) and short ( t _½ab = 2.33 ± 1.51 min) and the mean bioavailability was 91.0 ± 32.8%, although there was considerable interanimal variation. In some individuals, bioavailability was higher than 100%. This fact combined with the slower elimination phase after i.m. than after i.v. administration, could be related with enterohepatic recycling.     

Isolation and Culture of Ovine and Bubaline Small and Large Pre-antral Follicles: Effect of Cyclicity and Presence of a Dominant Follicle

Studies were conducted to examine the effects of the cyclicity and the presence of a dominant follicle (DF) in ovary on the recovery and in vitro growth of pre-antral follicles (PFs) in sheep and buffalo. Small pre-antral follicles (SPFs, 100–250 μm) and large pre-antral follicles (LPFs, 250–450 μm) were isolated from slaughterhouse ovaries in the breeding seasons by a mechanical and enzymatic method. The sheep and buffalo PFs were cultured in vitro for 6 and 15 days, respectively, and examined for their growth, survival and antrum formation rates and growth rates of oocytes in cultured pre-antral follicles. The follicles of the sheep and buffalo were recovered and cultured simultaneously within replicates. The recovery rates (number per ovary) of both SPFs and LPFs were significantly (p < 0.05) higher in cyclic ewes (SPFs: 22.0 ± 3.3 vs 12.1 ± 2.6 and LPFs: 16.0 ± 3.6 vs 9.2 ± 1.8) and buffaloes (SPFs: 9.2 ± 1.3 vs 4.1 ± 1.0 and LPFs: 10.3 ± 2.7 vs 5.4 ± 0.7) compared with those recovered from acyclic ones. Presence of a DF in ovary significantly (p < 0.05) reduced the recovery rates of LPFs in ewes (9.06 ± 2.7 vs 16.4 ± 3.8) but had no effect in buffalo. Cyclicity of animals or follicular dominance had no effects on in vitro growth, survival and antrum formation rates and growth rates of oocytes in cultured PFs of SPFs and LPFs in both sheep and buffalo. The in vitro growth, survival and antrum formation rates of LPFs and growth rates of oocytes in cultured LPFs were significantly (p < 0.05) higher than those observed in SPFs in both sheep and buffalo. The overall recovery and growth rates of the PFs were lower in buffaloes compared with ewes.     

Plasma pharmacokinetics and urine concentrations of meropenem in ewes

The pharmacokinetics of meropenem was studied in five ewes after single i.v. and i.m. dose of 20 mg/kg bw. Meropenem concentrations in plasma and urine were determined using microbiological assay method. A two-compartment open model was best described the decrease of meropenem concentration in plasma after an i.v. injection. The drug was rapidly eliminated with a half-life of elimination ( t _{1/2 β} ) of 0.39 ± 0.30 h. Meropenem showed a small steady-state volume of distribution [ V _d(ss)] 0.055 ± 0.09 L/kg. Following i.m. injection, meropenem was rapidly absorbed with a t _1/2ab of 0.25 ± 0.04 h. The peak plasma concentration ( C _max) was 48.79 ± 8.83  μ g/mL was attained after 0.57 ± 0.13 h ( t _max). The elimination half-life ( t _1/2el) of meropenem was 0.71 ± 0.12 h and the mean residence time ( MRT ) was 1.38 ± 0.26 h. The systemic bioavailability (F) after i.m. injection was 112.67 ± 10.13%. In vitro protein-binding percentage of meropenem in ewe's plasma was 42.80%. The mean urinary recoveries of meropenem over 24 h were 83% and 91% of the administered dose after i.v. and i.m. injections respectively. Thus, meropenem is likely to be efficacious in the eradication of many urinary tract pathogens in sheep.     

Improving the estimation of realized effective population sizes in farm animals

Computation of inbreeding rate (Δ F ) must consider that inbreeding is delayed with one generation with respect to the idealized population when addressed using individual inbreeding coefficients. The expression relating inbreeding in generation t with inbreeding rate F _t  = 1 – (1– ΔF ) ^t should be more correctly written in real animal populations as F _t  = 1 – (1– ΔF ) ^{t −1}, as changes in allele frequencies occur in the equivalent co-ancestries in the previous generation. This simple approach is tested on simulated and real pedigrees thus demonstrating that: (i) the adjusted individual increase in inbreeding becomes stable in populations under random mating while the unadjusted parameter does not; (ii) regression of the unadjusted parameter over generations in pedigrees under random mating is highly significant while after correction it is not significant; and (iii) the variance of the adjusted parameter is reduced with the generations.     

Effect of acepromazine or xylazine on tear production as measured by Schirmer tear test in normal cats

Objective  To evaluate the effect of acepromazine or xylazine on Schirmer tear test 1 results in clinically normal cats.
Animals  Sixteen healthy cross-breed cats.
Procedure  The animals were randomly divided into two groups of eight cats each. The first group was sedated with acepromazine alone (0.2 mg/kg) and the second group received only xylazine (2 mg/kg). All cats had Schirmer tear test (STT) readings taken prior to sedation and at 15 and 25 min postsedation.
Results  Sedation with acepromazine or xylazine in cats with normal pre-sedation STT 1 values caused a statistically significant decrease in mean values of tear production in both groups. In acepromazine group the mean ± SEM STT at T₁₅ and T₂₅ were 4.31 ± 0.98 ( P  < 0.001) and 5.18 ± 1.07 ( P  = 0.002). The post-treatment mean ± SEM values in xylazine group were 2.18 ± 0.97 ( P  < 0.001) and 2.62 ± 1.17 ( P  = 0.001) at 15 and 25 min respectively. Comparison between T₁₅ and T₂₅ in acepromazine group ( P  = 0.49) and xylazine group ( P  = 0.56) revealed no significant differences.
Conclusion  These observations indicate that both acepromazine or xylazine significantly reduced tear production in clinically normal cats. In cats, clinicians should measure STT values prior to utilizing acepromazine or xylazine as sedatives in order to accurately assess the results. Moreover, sterile ocular lubricant or tear replacement should be used as a corneal protectant during sedation with these drugs.     

Genetic relationship and diversity analysis of Indian water buffalo (Bubalus bubalis)

The water buffalo (Bubalus bubalis) is an important dairy animal on the Indian subcontinent and in Southeast Asian countries. The diversity and differentiation among 12 populations or breeds of buffalo were studied. Data were generated and analyzed from 527 animals belonging to 10 recognized breeds and 2 additional populations of Indian buffalo by using 22 microsatellite loci. Relationships among buffalo breeds and populations were estimated based on genetic distances. The Bayesian analysis grouped 12 populations into 8 distinctive clusters. Geographically close breeds clustered together, except for the Jaffarabadi and Murrah, which were not in geographic contiguity. The Mantel test revealed nonsignificant correlations between genetic and geographic distances. This supports the hypothesis that buffaloes have been domesticated at different places for specific purposes. The phylogenetic relationship based on microsatellite loci supported the breed classification based on body size. The Toda breed, which is considered to be endangered, had genotypes similar to those of the surrounding buffalo populations.     

摩拉水牛PRKAA2基因多态性与生长性状的关联分析

试验旨在探讨PRKAA2基因在摩拉水牛群体中的遗传多态性及其与生长性状的关联性,进而得到显著相关的遗传标记,为摩拉水牛的分子标记辅助选择提供理论依据。利用DNA测序法等技术进行候选基因PRKAA2外显子4及部分内含子3单核苷酸多态性（SNPs）检测,并采用生物信息学方法结合统计软件分析PRKAA2基因与摩拉水牛生长性状的关联性。结果显示,摩拉水牛PRKAA2基因中共检测到4个SNPs位点：c.462 G>A、IVS3.557 T>C、IVS3.560 C>T和IVS3.565 G>A,均包含3种基因型,且符合Hardy-Weinberg平衡。4个SNPs位点在水牛群体中均为中度多态,可以构建3种单倍型,T-C-G-G为优势单倍型,其中IVS3.560 C>T与c.462 G>A位点之间存在完全连锁不平衡,关联分析结果显示,单倍型H2与摩拉水牛体斜长和腰角宽呈显著相关（P<0.05）。c.462 G>A位点与摩拉水牛体高、十字部高、尻长、坐骨端宽和腰角宽呈显著相关,其中GG和AA基因型个体的体高和十字部高均显著高于GA基因型个体,GG基因型个体的尻长显著高于GA和AA基因型个体,AA基因型个体的坐骨端宽和腰角宽显著高于GA基因型个体（P<0.05）;IVS3.557 T>C位点与摩拉水牛的生长性状指标未达到显著相关（P>0.05）;IVS3.560 C>T位点与摩拉水牛体高、十字部高和尻长呈显著相关,其中CC和TT基因型个体的体高和十字部高均显著高于CT基因型个体,CC基因型个体的尻长显著高于CT基因型个体（P<0.05）;IVS3.565 G>A位点与体斜长呈显著相关,其中GG和GA基因型个体的体斜长显著高于AA基因型个体（P<0.05）。综上,PRKAA2基因的c.462 G>A、IVS3.560 C>T和IVS3.565 G>A位点对摩拉水牛部分生长性状均有显著影响,可作为摩拉水牛品种早期选育的候选基因和分子辅助标记。