松辽黑猪和长白猪催乳素受体基因第10外显子多态性与繁殖性状的关联分析

试验旨在探讨催乳素受体(prolactin receptor,PRLR)基因遗传多态性及其与松辽黑猪、长白猪繁殖性状的关联性。采用PCR-RFLP方法对松辽黑猪和长白猪PRLR基因第10外显子进行了多态性检测,并运用最小二乘法分析各基因型对母猪繁殖性能的遗传效应。结果表明,两个猪种中都存在A、B 2个等位基因和AA、AB、BB 3种基因型,松辽黑猪和长白猪等位基因A的频率分别0.551和0.578,A为优势基因。两个猪种的AA基因型个体的断奶仔猪数、乳头数都显著高于其他两个基因型(P＜0.05),初产松辽黑猪AA基因型个体的TNB、NBA显著高于其他基因型(P＜0.05),对其他繁殖性状影响不显著,但呈现出AA＞AB＞BB的趋势。     

A Comparative Study on the Carcass and Meat Quality Traits of Jiangquan Black Pig and DLY Pig

To further understand the germplasm characteristics of Jiangquan Black pig,100 kg Jiangquan Black pigs (10 barrows, 10 sows) and 10 Duroc×Landrace×Yorkshire (DLY) pigs under the same conditions were randomly selected to analyse the slaughter performance and meat quality according to related rules. After slaughtered,the carcass index (carcass weight, loin muscle area, carcass length and backfat thickness etc), meat quality physical indicators (pH, meat color, marbling, drip loss etc), gerneral chemical index (the contents of crude protein, intramuscular fat, moisture and ash etc) and the proportion of amino acids in Jiangquan Black pig were analyzed. The results showed that, Jiangquan Black pig carcass lean meat rate was relatively low, the loin muscle area was reduced by about 30% (P<0.05) compared with DLY pig, while the backfat thickness was higher than that of DLY pig (P<0.05). The meat color, marbling, drip loss, cooking loss, water loss rate index and intramuscular fat content of Jiangquan Black pig were significantly better than that of DLY pigs (P<0.05), the rate of water loss and drip loss and cooking loss were decreased by 4.0%, 3.1% and 2.7%, intramuscular fat content increased by 150.0%. Therefore, The lean yield of Jiangquan Black pig was less than DLY pig, but water retention property, intramuscular fat content, meat color, marbling, meat quality indexes were significantly better than that of DLY pig. In conclusion, Jiangquan Black pig could provide good quality pork for consumers.     

江泉黑猪与杜长大三元杂交猪屠宰性能及肉质性状比较分析

为了解江泉黑猪的产肉和肉质特性,本试验选取在相同条件下饲养的100 kg左右的江泉黑猪阉公猪、母猪和杜长大三元杂交猪各10头按照相关规定进行屠宰和肉品质测定。对两种猪胴体指标（胴体重、胴体长、眼肌面积、背膘厚等）、肉质的物理指标（pH、肉色、大理石纹、滴水损失等）、常规化学指标（粗蛋白质、肌内脂肪、水分、灰分含量等）及江泉黑猪氨基酸含量进行了测定。结果表明,与杜长大三元杂交猪相比,江泉黑猪胴体瘦肉率较低,眼肌面积降低了30%左右（P<0.05）,而胴体背膘厚较高（P<0.05）。江泉黑猪在肉色、大理石纹、滴水损失、失水率、烹饪损失和肌内脂肪含量等指标上均显著优于杜长大三元杂交猪（P<0.05）,其中滴水损失、失水率和烹饪损失分别降低了约4.0%、3.1%和2.7%,肌内脂肪含量提高了约150.0%。由此可见,江泉黑猪在瘦肉产量上低于杜长大三元杂交猪,但在保水性、肌内脂肪含量、肉色、大理石纹等肉质指标上明显优于杜长大三元杂交猪,可为消费者提供品质优良的猪肉。     

Cloning and Tissue Expression Analysis of PDK4 and FGF10 Genes in Guizhou Congjiang Xiang Pig

The objective of this study was to clone PDK4 and FGF10 genes, and investigate the expression level of PDK4 and FGF10 genes mRNA in different tissues of Large White pig and Congjiang Xiang pig. The PDK4 and FGF10 genes were cloned by RT-PCR and analyzed by bioinformatics, the relative expression of PDK4 and FGF10 genes were detected by Real-time PCR. The results showed that the coding region of PDK4 gene was 1 224 bp, encoding 407 amino acids; The coding region of FGF10 gene was 636 bp and encoded 211 amino acids. The homologies of nucleotide sequences of PDK4 gene with sheep, horse and human were 93%, 92% and 91%,respectively. The homologies of nucleotide sequences of FGF10 gene with sheep, cattle, human and mouse were 94%,93%, 93% and 90%, respectively. The phylogenetic tree of PDK4 gene showed that the genetic relationship of Congjiang Xiang pig, cattle and sheep were very close, the phylogenetic tree of FGF10 gene indicated that the genetic relationship of Congjiang Xiang pig, cattle, sheep, human and macaque were very close, but the genetic relationship of Congjiang Xiang pig, rat and chicken were far away. Real-time PCR results showed that, in different tissues of Congjiang Xiang pig,PDK4 gene expression in kidney tissue was higher than other tissues, with a higher expression in stomach and adipose as well,FGF10 gene expression in stomach tissue was higher than other tissues, with a higher expression in kidney and adipose as well, but both of PDK4 and FGF10 genes expression were the lowest in longissimus dorsi. In different tissues of Large White pig, both of PDK4 and FGF10 genes were expressed the highest in adipose than other tissues, PDK4 gene expression in longissimus dorsi was the lowest, while the FGF10 gene expression the lowest in heart. This study successfully cloned the PDK4 and FGF10 genes of Large White pig and Congjiang Xiang pig,and detected the relative expression of PDK4 and FGF10 genes in different tissues of Large White pig and Congjiang Xiang pig, and also provided scientific basis for further study on regulation of PDK4 and FGF10 genes on lipid metabolism and deposition.     

贵州从江香猪PDK4、FGF10基因的克隆及组织表达分析

试验旨在克隆获得PDK4、FGF10基因,并研究大白猪与从江香猪不同组织中PDK4、FGF10基因mRNA的表达差异。采用RT-PCR分别克隆从江香猪PDK4、FGF10基因并进行生物信息学分析,利用实时荧光定量PCR技术检测PDK4、FGF10基因在大白猪和从江香猪不同组织中mRNA的相对表达量。结果显示,从江香猪PDK4基因的编码区全长1 224 bp,编码407个氨基酸;FGF10基因的编码区全长636 bp,编码211个氨基酸。经BLAST软件进行同源性比对,发现从江香猪PDK4基因与羊、马、人的核苷酸序列同源性分别为93%、92%和91%;FGF10基因与羊、牛、人、鼠的核苷酸序列同源性分别为94%、93%、93%和90%。由PDK4基因系统进化树可知,从江香猪与牛、绵羊亲缘关系较近;由FGF10基因系统进化树可知,从江香猪与绵羊、牛、人、猕猴亲缘关系较近,与小鼠和鸡亲缘关系较远。实时荧光定量PCR结果显示,在从江香猪不同组织中,PDK4基因在肾脏中的表达量最高,在胃和脂肪中表达量较高,FGF10基因在胃中表达量最高,在肾脏和脂肪中表达量较高,两个基因在背最长肌中的表达量均最低;在大白猪的不同组织中,PDK4、FGF10基因在脂肪中的表达量均最高,PDK4基因在背最长肌中的表达量最低,而FGF10基因在心脏中表达量最低。本试验成功克隆了从江香猪PDK4、FGF10基因,并检测了其在大白猪与从江香猪不同组织中的表达,为进一步研究PDK4、FGF10基因在脂质代谢及脂肪沉积等方面的调控作用提供科学依据。     

Studies on the carriage and transmission of porcine reproductive and respiratory syndrome virus by individual houseflies (Musca domestica)

The objectives of the study were to determine the site of porcine reproductive and respiratory syndrome virus (PRRSV) in individual houseflies, to assess whether an individual housefly could transmit PRRSV to a susceptible pig, and to compare the ability of PCR, virus isolation and a pig bioassay to detect PRRSV in houseflies. In the first experiment 26 houseflies were fed on a pig infected experimentally with PRRSV; 13 were processed as a whole fly homogenate, while an exterior surface wash and a gut homogenate were collected from the other 13. Infectious PRRSV was recovered from nine of the whole fly homogenates, 12 of the gut homogenates and one of the exterior surface washes. In the second experiment, two of 10 individual houseflies, which had fed on an infected pig, transmitted PRRSV to a susceptible pig in a controlled manual transmission protocol. In the third experiment, single flies or pools of 30 flies were immersed in different concentrations of a PRRSV inoculum, then tested by PCR, virus isolation and bioassay. The virus was detected at a concentration of 10(1) TCID50/ml by PCR, 10(2) TCID50/ml by the bioassay and 10(3) TCID50/ml by virus isolation.     

Methicillin‐Resistant Staphylococcus aureus (MRSA) ST398 in Pig Farms and Multispecies Farms

During the last few years, methicillin‐resistant Staphylococcus aureus (MRSA) ST398 has been isolated frequently from livestock, especially from pigs and to a lesser extent from cattle and poultry. To gain insight into the distribution of this bacterium in pig farms versus multispecies farms, 30 Belgian farms (10 pig, 10 pig/poultry and 10 pig/cattle farms) were screened for the presence of MRSA. On each farm, 10 nasal swabs were taken from pigs. When present, cattle (n = 10) were sampled in the nares and poultry (n = 10) in the nares, earlobes and cloaca. A selection of the obtained isolates were further characterized using multilocus sequence typing (MLST), spa typing, SCCmec typing, pulsed field gel electrophoresis (PFGE), multiple‐locus variable‐number tandem repeat analysis (MLVA) and antimicrobial susceptibility testing. On 26 of 30 farms, MRSA was isolated from pigs. Furthermore, MRSA was also isolated from poultry and cattle on one pig/poultry and five pig/cattle farms, respectively. All tested MRSA isolates belonged to ST398. Eight spa types (t011, t034, t567, t571, t1451, t2974, t3423 and t5943) were detected, among which t011 predominated. SCCmec cassettes type IVa and V were present in 20% and 72% of the isolates, respectively. When combining the results of the two remaining typing methods, PFGE and MLVA, eighteen genotypes were obtained of which one genotype predominated (56% of the positive farms). All MRSA isolates were resistant to tetracycline. Resistance to trimethoprim, aminoglycosides, macrolides, lincosamides, fluoroquinolones and chloramphenicol was also observed. In conclusion, there was no effect of the farm type on the MRSA status of the pigs. A statistically significant difference was observed when comparing the pig/poultry or the pig/cattle MRSA status on the multispecies farms. Additionally, a wide variety of MRSA ST398 strains was found within certain farms when combining different typing methods.     

Microbial contamination of dust in minimum-morbidity housings for pig rearing]

The range of the microbial contamination of 1 g of dust in minimum-morbidity pig rearing houses is from 2.9 . 10(7) to 7.6 . 10(9) in the microbes growing on meat-peptone agar (MPA), from 4.6 . 10(7) to 4.6 . 10(8) in moulds growing on the Czapek-Dox agar, from 1.4 . 10(2) to 7.0 . 10(4) in the total number of microbes growing on End's agar, and from 3.1 . 10(2) to 5.7 . 10(3) in the typically lactose-positive microbes growing on End's agar. The numbers of microbes in 1 g of dust in the conventional pig houses are close to the numbers found in pig rearing houses with minimum morbidity. However, in the moulds on the Czapek-Dox agar, the range of the microbial contamination of dust in the minimum-morbidity pig houses was much higher. The highest level of the contamination of dust with microbes was found on all types of media from houses with continual operation. It was revealed by the control of the effectiveness of disinfection in the minimum-morbidity piglet rearing section, operated on the all-in all-out basis, that in a half of the cases studied the disinfection had low effectiveness. The total number of microbes per 1 sq. cm was lower than required on the basis of research published in literature, but the occurrence rate of positive smears with lactose-positive microbes after disinfection was higher in percentual expression.     

荣昌猪和两种小型猪遗传背景的微卫星分析

荣昌猪是我国优良地方品种,巴马香猪和贵州小型香猪为我国独特小型猪品系。本研究采用美国猪基因组协作计划推荐的19个微卫星标记对荣昌猪、荣昌猪B系、巴马香猪、贵州小型香猪及外来猪种大白猪进行了遗传学检测和分析。结果表明:19个微卫星位点在群体中均表现为多态,每位点等位基因数10~23个。5个猪种中荣昌猪B系的遗传多样性最高,荣昌猪次之;2种小型猪的遗传多样性较低,其中巴马香猪的等位基因数(121个)略大于贵州小型香猪(114个),但其遗传多样性指数(平均期望杂合度0.6535±0.0347)小于贵州小型香猪(0.6919±0.0227),反映了巴马香猪较低的基因杂合度和较小的遗传变异;大白猪的遗传多样性最低。从品系的共有等位基因来看,荣昌猪B系基因背景组成中其亲本品系荣昌猪所占比例(30.22%)要大于大白猪(24.37%);大白猪和其他4个猪群体的共享等位基因数均较低(21.24%~25.12%);巴马香猪和贵州小型香猪之间共有等位基因数最高(45.06%)。采用基于遗传距离的NJ法和基于基因频率的最大似然法进行系统聚类,除了大白猪明显为独立分支及荣昌猪B系表现出较远的聚类关系外,其他3个猪种间的聚类有所不同。     

荧光定量PCR在高致病性猪蓝耳病检测上的应用

应用实时荧光定量RT-PCR方法对上海松江区8个生猪重点养殖地区的60份样品（血液样品50份、组织病料10份）,以及10个规模化猪场的100份样品（血液样品81份、组织病料19份）进行高致病性猪蓝耳病的检测,结果均为阴性,表明该区猪群中没有高致病性猪蓝耳病的隐性感染和健康带毒。同时,熟悉并完善了高致病性猪蓝耳病的荧光定量RT-PCR检测技术。     

SNP技术在猪肉溯源过程中的应用

为筛选可用于梅山猪和申农猪肉质溯源的SNP位点标记,采用PCR-RFLP方法检测13个基因在梅山猪和申农猪群体内14个SNP位点的多态性,以期寻找多态性丰富的SNP位点,通过再次采集样品,进行溯源验证试验,最终确定可用于梅山猪和申农猪肉溯源的SNP位点组合。结果表明：MMP19基因、GRN基因、NR4A1基因的SNP位点和PSMB10基因的2个SNP位点杂合度（H值）均大于0.30,符合肉质溯源要求。可根据MMP19、PSMB10、NR4A1和GRN 4个基因共5个SNP位点编制用于检测梅山猪和申农猪肉质溯源的条形码。     

三江白猪相对于亲本特异性条带的筛选

本试验是出于三江白猪的保种计划所做的研究。利用RAPD技术,在前期研究的基础上,利用已筛选出的15条10bp的随机引物,在已经优化的RAPD反应条件基础上,对三江白猪及其亲本长白猪与东北民猪组织DNA进行PCR分析。从中筛选出三江白猪相对于亲本的特异性条带,找出三江白猪相对于亲本的基因组变异情况,为三江白猪种间特异性条带的确定奠定基础。     

应用分子标记AFLP建立不同猪种间遗传关系

作者首先在实验室建立了猪的AFLP分析体系。经过筛选,利用16对引物组合构建了中国6个常见猪品种中共计20个样本的AFLP分析图谱,通过统计分析,得到了6个猪品种的遗传距离,并由此构建了聚类分析图。在此基础上,得到了一些品种特异性的片段,这些片段为进一步寻找这些品种特异性的分子标记提供了可能。     

Production constraints of smallholder pig farms in agro-ecological zones of Mpumalanga,South Africa

South African pig sector is a contributor to the agricultural industry. A study was conducted to identify the production constraints and compare the management practices in smallholder pig farms in Mpumalanga, South Africa. A total of 220 selected smallholder pig farmers were interviewed. Smallholder pig farming was predominated by male (64 %), age above 50 years (54 %), black Africans (98.6 %), and three quarters of the smallholder farmers were poor to just below average. Majority (80 %) have no pig husbandry training, while only 33 % received assistance from government’s Agricultural Department. In terms of stock, mixed breeds (89 %) from exotic pigs were mostly kept and majority (87 %) of the farmers kept ≤10 sows in their herds. Many farmers (75 %) engaged in risky behavior of buying auctioned-sourced boars, free-range boars, and untested boars from neighbors and relatives. Few (17 %) farmers practiced vaccination and only 10 % kept farm records. Majority of the responses on pre-weaning mortality (50 %) and post-weaning mortality (90 %) were within acceptable range of 1–10 and 1–5 % mortality rates, respectively. The lead causes of mortality were weak piglets and crushing (46 %), diarrhea (27 %), poor management knowledge (19 %), and malnutrition (16 %). Agricultural training and government incentives will facilitate improved productivity in smallholder pig farming.     

安徽地方猪种染色体Ag-NORs分析研究

对安徽省皖南花猪、皖南黑猪、定远猪、圩猪、安庆六白猪等五个地方猪种Ag-NOR s多态性进行了分析,结果显示Ag-NOR s均数分别为3.69、3.67、3.17、3.73、3.50,众数为4;2个引进品种大约克夏和长白Ag-NOR s均数分别为2.14、2.11,众数为2。Ag-NOR s均定位于10号和8号染色体次缢痕区。从研究结果可以推断皖南花猪和圩猪的品种纯度较高,聚类分析可将5个地方猪种分为2类:皖南花猪、皖南黑猪和圩猪为第1类,定远猪和安庆六白猪为第2类。此外5个地方猪种和2个引进品种Ag-NOR s多态性还为中国猪种起源于亚洲野猪,欧洲猪种起源于欧洲野猪提供了进一步的依据。     

Survival of Aujeszky's disease virus in infected pig slurry

It has been demonstrated that after experimental infection of pig slurry from the space under the slatted floor (infection dose of 10(6)PFU per ml), the Aujeszky's disease virus (ADV) survived for 72 hours at the temperature of 15 degrees C and at pH 6.5, but was inactivated after 96 hours. When technologically treated pig slurry from the storage tanks was saturated with water and infected with ADV at the dose of 10(5)PFU per ml, the virus survived for 23 days when kept at 15 degrees C and 4 degrees C and at pH 6.8, but was inactivated under the same conditions after 30 days. When the infective ADV dose in the technologically treated pig slurry in the storage tanks was reduced to 10(4)PFU per ml, the virus survived 16 days at +4 degrees C and pH 7.0 and 8.0 but was inactivated within 23 days after infection.     

Occurrence of Cryptosporidium and Giardia in suckling piglets in Norway

Faecal samples from 684 litters of suckling piglets from 100 indoor swine herds from all regions of Norway were examined for the presence of Cryptosporidium and Giardia, using sucrose gradient flotation concentration and immunofluorescent staining. Thirty-one (31%) herds and 57 (8.3%) litters tested positive for Cryptosporidium, while 10 (1.5%) litters in 10 different herds (10%) tested positive for Giardia. Molecular characterisation of nine Cryptosporidium isolates demonstrated both C. suis and Cryptosporidium sp. pig genotype II. There was significantly more diarrhoea among the Cryptosporidium positive piglets than among the Cryptosporidium negative piglets. Diarrhoea was not observed amongst litters in which Cryptosporidium sp. pig genotype II was found. There was a significant difference in the Cryptosporidium prevalence between litters from different geographical areas of Norway. This study demonstrates that both Cryptosporidium (C. suis and Cryptosporidium sp. pig genotype II) and Giardia infections are prevalent among suckling piglets in Norway.     

香猪SNP位点rs80894395的多态性及其与体尺指标之间的关联研究

为了探究影响香猪生长发育的功能基因,本试验利用Illumina Porcine SNP60K芯片筛选出N-乙酰氨基半乳糖转移酶2(polypeptide N-acetylgalactosaminyltransferase 2,GALNT2)基因内含子10中一个与腹围呈弱相关的单核苷酸多态性(single nucleotide polymorphism,SNP)位点rs80894395。在此基础上应用等位基因特异性PCR(allele-specific PCR,AS-PCR)技术检测520头雌性香猪群体中该位点的基因型,并与43头大白猪、36头柯乐猪相比较。结果表明,从3个猪群体中检测到rs80894395位点的CC、CT和TT3种基因型;其中香猪群体以CT为优势基因型,且基因型与香猪的体重、体长和腹围呈弱的正相关;大白猪群体以CC基因型为主,T等位基因频率较香猪极显著减少(P<0.01);柯乐猪群体中CC与CT基因型频率相等,与香猪和大白猪的基因频率间的差异不显著(P>0.05)。生物信息学分析表明,rs80894395位点可能影响GALNT2基因的剪接过程。rs80894395与香猪的生长发育可能有一定的联系。     

Evaluation of mosquitoes, Aedes vexans, as biological vectors of porcine reproductive and respiratory syndrome virus

The objective of this study was to determine whether mosquitoes, Aedes vexans (Meigen), could serve as biological vectors of porcine reproductive and respiratory syndrome virus (PRRSV). Specifically, the study assessed the duration of viability and the site of PRRSV within mosquitoes, and evaluated whether PRRSV could be transmitted to a susceptible pig by mosquitoes following a 7- to 14-day incubation period after feeding on an infected pig. For the first experiment, a total of 100 mosquitoes were allowed to feed on a pig, experimentally infected with PRRSV (day 7 post-inoculation) and were then maintained alive under laboratory conditions. A set of 10 mosquitoes were collected at 0 hour (h), 6 h, 12 h, 24 h, 48 h, 72 h, 5 days (d), 7 d, 10 d, and 14 d post-feeding (pf). Samples of exterior surface washes, salivary glands, thorax carcasses, and gut homogenates were collected from each set of mosquitoes, and tested for PRRSV. Infectious PRRSV was detected by polymerase chain reaction and swine bioassay only from the gut homogenates of mosquitoes collected at 0 h and 6 h pf. For the second experiment, a total of 30 mosquitoes were allowed to feed on a pig, experimentally infected with PRRSV and the mosquitoes were then maintained under laboratory conditions. On each of day 7, 10, and 14 pf, a set of 10 mosquitoes were allowed to feed on a susceptible pig. Transmission of PRRSV to susceptible pigs did not occur, and PRRSV was not detected from the mosquitoes. These findings indicate that mosquitoes are not likely to serve as biological vectors of PRRSV.