Distribution and Genotypic Characterization of Campylobacter jejuni Isolated from Poultry in Split and Dalmatia County,Croatia

Consumption of poultry contaminated with Campylobacter jejuni has been recognized worldwide as the leading cause of campylobacteriosis. Therefore, the aim of our study was to investigate the prevalence and genotype diversity of Campylobacter jejuni in poultry meat intended for consumption in Split and Dalmatia County, which is the second biggest County in Croatia. Furthermore, we also wanted to discover possibly stable clones of C. jejuni appearing in different samples and periods of time, which would indicate their ability to persist in or adapt to poultry. In the period from March 2008 until June 2010, 834 samples of poultry from various sources were examined using a surface swab technique. Isolation of C. jejuni was performed by Preston broth and Karmali agar. Identification of the isolates was carried out using biochemical tests. C. jejuni was found in 84 of 574 chicken samples (14.6%) and in nine of 260 samples of turkey (3.5%). Pulse‐field gel electrophoresis (PFGE) was used to analyse 61 obtained isolates using SmaI and KpnI. Of 22 different macrorestriction profiles (MRP) that were found, five were detected in poultry from both different locations and periods of time. Samples from 11 locations were found to be contaminated with more than two different genotypes of C. jejuni. Interestingly, the same MRP were found both in poultry declared to be of domestic origin and in the poultry imported from abroad. The prevalence of C. jejuni in poultry samples was in accordance with previously reported results. Genotypic analysis indicated that the population of C. jejuni in Split and Dalmatia County was diverse and that multiple strains of C. jejuni could be found in the same poultry samples. Furthermore, the same genotypes were identified from the samples obtained from different locations and periods of time, which could support the theory of a global existence of certain MRP that are able to persist in or adapt to poultry.     

绵羊DUSP6基因多态性及其与产羔数关联分析

文章旨在探究绵羊DUSP6基因g.125589716G>A、g.125587728C>T、g.125589714C>T、g.125589006G>A四个位点多态性及其与产羔数之间的关系,以期找到与绵羊高繁殖力相关的分子标记。利用全基因组重测序结合Sequenom MassARRAY誖SNP技术对多羔绵羊品种(小尾寒羊、湖羊、策勒黑羊)和单羔绵羊品种(滩羊、苏尼特羊、萨福克羊、草原型藏羊)DUSP6基因上述4个多态位点进行检测,并与小尾寒羊产羔数进行关联分析。结果表明:绵羊g.125589716G>A位点在多羔绵羊品种中存在AA、GA、GG三种基因型,在单羔绵羊品种中存在GA、GG两种基因型;g.125587728C>T、g.125589714C>T位点在单、多羔绵羊品种中均存在CC、CT、TT三种基因型;g.125589006G>A位点在单、多羔绵羊群体中均只存在GA、GG两种基因型。绵羊DUSP6基因g.125589716G>A位点基因型频率在单、多羔绵羊品种之间差异显著(P<0.05),等位基因频率在单、多羔绵羊品种间差异极显著(P<0.01);g.125587728C>T、g.125589714C>T位点基因型频率和等位基因频率在单、多羔绵羊品种间差异均达极显著水平(P<0.01);g.125589006G>A位点基因型频率和等位基因频率在单、多羔绵羊品种间差异不显著(P>0.05)。关联分析表明,DUSP6基因各SNPs多态性与小尾寒羊不同胎次产羔数之间无显著关联(P>0.05)。综上说明,绵羊DUSP6基因g.125589716G>A、g.125587728C>T、g.125589714C>T以及g.125589006G>A等4个位点的多态性与小尾寒羊各胎次产羔数之间均无显著关联(P>0.05),不适用于小尾寒羊多羔性状选育。     

Real-time PCR genotyping assay for feline erythrocyte pyruvate kinase deficiency and mutant allele frequency in purebred cats in Japan

Erythrocyte pyruvate kinase (PK) deficiency is an inherited glycolytic erythroenzymopathy caused by mutations of the PKLR gene. A causative mutation of the feline PKLR gene was originally identified in Abyssinian and Somali cats in the U.S.A. In the present study, a TaqMan probe-based real-time PCR genotyping assay was developed and evaluated for rapid genotyping and large-scale screening for this mutation. Furthermore, a genotyping survey was carried out in a population of four popular purebred cats in Japan to determine the current mutant allele frequency. The assay clearly displayed all genotypes of feline PK deficiency, indicating its suitability for large-scale survey as well as diagnosis. The survey demonstrated that the mutant allele frequency in Abyssinian and Somali cats was high enough to warrant measures to control and prevent the disease. The mutant allele frequency was relatively low in Bengal and American Shorthair cats; however, the testing should still be carried out to prevent the spread of the disease. In addition, PK deficiency should always be considered in the differential diagnosis of anemia in purebred cats in Japan as well as worldwide.     

Genotyping of Bacillus anthracis Isolated from Croatia and Bosnia and Herzegovina

Anthrax is a serious disease caused by Bacillus anthracis. Humans can become infected by handling products from infected animals, by breathing spores and rarely by eating undercooked meat from infected animals. The genome of B. anthracis is highly monomorphic and thus shows very low DNA sequence variation. We analysed the molecular characteristics of 12 B. anthracis isolates from outbreaks in Croatia and Bosnia and Herzegovina, which have occurred during the past 10 years along with two vaccine strains. Genotyping system based on variable‐number tandem repeat analysis at six loci revealed that six isolates belong to genotype from the A1.a cluster whilst six isolates relate to the B2 cluster, compared to 89 previously described genotypes. The distribution of two evolutionarily distant clusters suggests an introduction of B. anthracis to this area in at least two separate events.     

全基因组SNP分型技术在畜禽遗传育种研究中的应用

随着畜禽资源分子鉴定、物种进化、全基因组育种等热点领域的逐渐兴起,准确的全基因组SNP分型成为了畜禽基因组研究的关键。基因芯片、重测序、简化基因组测序及靶向捕获测序等全基因组SNP分型技术已广泛应用于畜禽基因组研究中。本文概述了全基因组SNP分型技术的原理及其在全基因组关联分析、选择信号分析和畜禽遗传资源背景分析等方面的应用,以期为畜禽基因组研究和育种应用提供借鉴和参考。     

Isolation,Identification and Genotyping of Pasteurella multocida from Fowl Cholera

In order to determine the suspected avian cholera pathogen and geneotypes, bacteria isolation technology was used to isolate and culture pathogenic bacteria, the isolated bacteria were identified by traditional methods and molecular biological methods. Geneotyping of the isolated bacteria were determined using PCR technology and gene sequence analysis. The results demonstrated that the isolated bacteria had typical culture characteristics of Pasteurella multocida (Pm),and the morphology of the colony, the characteristics of cell staining, the physiological and biochemical characteristics were consist with Pm; 457 bp fragment was amplified by PCR. Geneotyping results showed that only A primer amplified the target gene fragment of 1 050 bp, sequence analysis also showed that the isolated bacteria shared 97.6% to 100.0% sequence homology with reference strains, and phylogenetic tree analysis showed that the isolated bacteria and A type Pm were in a branch.The experimental results indicated that the isolated bacteria was identified as capsular serotype A of Pm, the results provided reference for the prevention and control of fowl cholera.     

猪、鸡源分离粪肠球菌的核糖体分型及耐药性分析

对上海地区1个规模化养猪场和1个规模化养鸡场分离鉴定的40株粪肠球菌,采用微量肉汤稀释法对12种抗菌药物进行药敏试验,采用Riboprinter^®微生物基因指纹鉴定系统检测分离菌株的基因指纹图谱,分别检测分离菌株中的核糖体型和耐药性。结果表明,在40株粪肠球菌中,80%以上为多重耐药菌株,有4株猪源粪肠球菌对万古霉素耐药,其中有2株表现为对万古霉素高度耐药（MIC>64 mg/L）,且对链霉素和庆大霉素同时表现为高度耐药（MIC>2 048 mg/L）。从耐药表型和核糖体分型共同分析粪肠球菌的耐药性,发现多重耐药性严重,同一核糖体型菌株的耐药表型并非完全一致。     

绵羊SPP1和PLCB3基因多态性与产羔数的关联分析

试验旨在探究SPP1基因g.36651870T>C位点多态性、PLCB3基因g.41871219T>C位点多态性与绵羊产羔数之间的关系,以期寻找绵羊产羔数性状相关的分子标记.利用全基因组重测序结合Sequenom MassARRAY?SNP技术对多羔羊(小尾寒羊、湖羊、策勒黑羊)和单羔羊(滩羊、苏尼特羊、萨福克羊和草原...     

病原体溯源技术研究进展及在寄生虫上的应用

病原体是导致人和动植物感染疾病的根源,带来一系列的安全隐患。病原体溯源研究对于基因型鉴定、分类地位准确化、流行病学调查及疾病的防控具有重大意义。近年来,分子生物学技术已被广泛用于病原溯源研究。作者主要对病原体溯源技术的最新研究进展及其在寄生虫上的应用作一综述。