全基因组测序技术在猪源沙门氏菌血清分型和耐药性分析中的应用

【目的】为研究全基因组测序技术在猪源沙门氏菌血清型和耐药性检测方面的应用能力。【方法】以60株猪源沙门氏菌为研究对象,应用传统玻片凝集法和微量肉汤稀释法对其进行血清分型和耐药性检测,并提取所有菌株核酸进行全基因组测序组装,利用SeqSero2和ResFinder4在线预测每株菌的血清型和耐药基因,然后与传统方法进行比较分析。【结果】血清凝集试验共鉴定出8种沙门氏菌血清型,而全基因组测序鉴定出9种血清型,二者分型结果的符合率为90.0%;对11种抗菌药物的药敏试验显示：沙门氏菌对四环素（73.3%）的耐药率最高,其次为氨苄西林（66.7%）、磺胺异噁唑（66.7%）和复方新诺明（53.3%）,基于全基因组预测的恩诺沙星、美罗培南、头孢噻呋和阿奇霉素的耐药性与药敏试验结果完全一致,对氨苄西林、磺胺异噁唑、四环素预测的结果符合率也均大于90.0%。【结论】全基因组测序分析的沙门氏菌血清型和耐药性与常规检测方法的符合率较高,且具有操作简便的优势,可以作为分析沙门氏菌血清型和耐药性的有效方法。     

驴源马流产沙门菌的耐药表型及毒力基因检测

为了解驴源马流产沙门菌临床分离株的耐药表型及毒力基因的携带情况,本试验通过K-B药敏纸片扩散法对9株马流产沙门菌进行11种临床常用抗菌药物的敏感性测试,采用PCR方法检测与沙门菌致病性相关的10种毒力基因。结果显示:9株驴源马流产沙门菌存在2种耐药表型,对链霉素耐药率达100%,对阿莫西林耐药率为33%;10种毒力基因中仅traT未被检出,invA、avrA、ssaQ、mgtC、sopB、spvC、spvR、pefA和misL的检出率均为100%。结果表明,驴源马流产沙门菌耐药谱较为单一,但携带的毒力基因为多样性组合。     

Antimicrobial stewardship in the dairy industry: responding to the threat of antimicrobial resistance

Dairy Australia is the national service body for the Australian dairy industry. Its role is to help farmers adapt to a changing operating environment and achieve a profitable, sustainable dairy industry. Although the use of antibiotics in Australian agriculture is relatively low in global terms, Dairy Australia recognises important drivers for continuous improvement in antimicrobial stewardship (AMS). Dairy Australia’s first strategic priority is to support profitable farms. This priority has driven the development of a range of on‐farm change management programs in the animal health and welfare fields to optimise the unit cost of production and dairy cattle welfare. Dairy Australia’s third strategic priority is to further develop a ‘trusted dairy industry’. Previous and current work under these two strategies position the dairy industry favourably with respect to confronting the challenge of antimicrobial resistance (AMR) and supporting sound AMS with the guiding principle of ‘as little as possible, as much as necessary’. However, given an incomplete but ongoing threat of AMR, more work is needed. Supported by Dairy Australia, the dairy industry has developed an antimicrobial use strategy aligning with the Australian Animal Sector National AMR Plan 2018.     

Prevalence and antimicrobial resistance of Campylobacter and Salmonella in layer flocks in Honshu,Japan

Campylobacter and non-typhoidal Salmonella are the major causes of bacterial gastrointestinal infections in humans. Although antimicrobial therapy is typically not recommended in many cases of these infections, it may be life-saving in patients with severe symptoms. Since chicken eggs and meat derived from layers are destined for human consumption, we investigated the prevalence and antimicrobial resistance of these two bacterial genera in 82 layer flocks at chicken processing plants in Honshu, Japan. Campylobacter was isolated from 77 flocks (93.9%). Resistance to ampicillin, tetracycline, and ciprofloxacin was documented in 42.3 (30/71), 16.9 (12/71), and 14.1% (10/71) of Campylobacter jejuni, respectively. Multilocus-sequence typing identified ST4389 and ST5262 as the most frequent C. jejuni sequence types. In C. coli, resistance to ampicillin, tetracycline, and ciprofloxacin was found in 20.0 (7/35), 20.0 (7/35), and 25.7% (9/35), respectively. The most frequent sequence type in C. coli was ST8292. Erythromycin resistance was not observed among Campylobacter species. Salmonella was isolated from 14 flocks (17.1%). The two most frequent serovars were Salmonella Corvallis and S. Braenderup. Neither S. Enteritidis nor S. Infantis were isolated. Streptomycin resistance was observed in six isolates (26.1%), and all isolates were susceptible to cefotaxime and ciprofloxacin. Thus, chicken eggs and meat derived from layers are possible sources of these bacterial infections in humans. The antimicrobial susceptibility of these isolates was maintained, reflecting restrictions on the use of antimicrobial agents on layers.     

肉鸡产业链中沙门菌流行情况及其耐药性

为了了解食源性病原沙门菌在肉鸡产业链中的流行情况及其耐药性现状。本研究从上海某肉鸡屠宰加工企业分别采集待宰肉鸡、屠宰胴体和市售鸡肉样品388、200、127份,并参照GB/T4789.4-2010和PCR法进行沙门菌的分离鉴定。结果显示,共获得沙门菌73株。待宰肉鸡、屠宰胴体和市售鸡肉分别获得沙门菌9、48和16株,分离率依次为2.32%、17.5%和8.66%。采用微量肉汤稀释法测定分离菌株对12种抗菌药物的敏感性。药敏结果显示,所有菌株至少耐两种药物。其中对复方新诺明和多西环素的耐药率最高,分别为100%和95.89%;头孢唑啉、阿莫西林/克拉维酸、氯霉素、氨苄西林、萘啶酸的耐药率也均超过了40%,而对头孢噻呋、头孢曲松、庆大霉素、美罗培南的耐药率相对较低。结果表明,该地区肉鸡源沙门菌的耐药情况较严重,亟需加强养禽业抗菌药物的合理应用;同时沙门菌流行情况也对改进肉鸡屠宰加工工艺,降低沙门菌的污染提出了要求。     

Prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma

Currently, antimicrobial-resistant staphylococci, particularly methicillin-resistant Staphylococcus pseudintermedius (MRSP), are frequently isolated from canine superficial pyoderma in Japan. However, little is known regarding the nasal prevalence of MRSP in pet dogs. Here, we determined the prevalence of antimicrobial-resistant staphylococci in nares and affected sites of pet dogs with superficial pyoderma. Of the 125 nares and 108 affected sites of pet dogs with superficial pyoderma, 107 (13 species) and 110 (eight species) staphylococci strains, respectively, were isolated. The isolation rate of S. pseudintermedius from pyoderma sites (82/110 strains, 74.5%) was significantly higher than that from nares (57/107 strains, 53.3%) (P<0.01). Notably, the prevalence of MRSP (18/57 strains, 31.6%) in nares was equivalent to that in pyoderma sites (28/82 strains, 34.1%). Furthermore, the phenotypes and genotypes of antimicrobial resistance in MRSP strains from nares were similar to those from pyoderma sites. Our findings revealed that the prevalence of antimicrobial-resistant staphylococci in the nares of pet dogs with superficial pyoderma is the same level as that in affected sites. Therefore, considerable attention should be paid to the antimicrobial resistance of commensal staphylococci in companion animals.     

Molecular serotyping and antimicrobial resistance profiles of Actinobacillus pleuropneumoniae isolated from pigs in South Korea

Background: Actinobacillus pleuropneumoniae (APP) causes porcine pleuropneumonia (PP).

Objective: Serotypes and antimicrobial resistance patterns in APP isolates from pigs in Korea were examined.

Methods: Sixty-five APP isolates were genetically serotyped using standard and multiplex PCR (polymerase chain reaction). Antimicrobial susceptibilities were tested using the standardized disk-agar method. PCR was used to detect β-lactam, gentamicin and tetracycline-resistance genes. The random amplified polymorphic DNA (RAPD) patterns were determined by PCR.

Results: Korean pigs predominantly carried APP serotypes 1 and 5. Among 65 isolates, one isolate was sensitive to all 12 antimicrobials tested in this study. Sixty-two isolates was resistant to tetracycline and 53 isolates carried one or five genes including tet(B), tet(A), tet(H), tet(M)/tet(O), tet(C), tet(G) and/or tet(L)-1 markers. Among 64 strains, 9% and 26.6% were resistance to 10 and three or more antimicrobials, respectively. Thirteen different antimicrobial resistance patterns were observed and RAPD analysis revealed a separation of the isolates into two clusters: cluster II (6 strains resistant to 10 antimicrobials) and cluster I (the other 59 strains).

Conclusion: Results show that APP serotypes 1 and 5 are the most common in Korea, and multi-drug resistant strains are prevalent. RAPD analysis demonstrated that six isolates resistant to 10 antimicrobials belonged to the same cluster.     

乌鲁木齐地区不同健康状态和饲养环境中宠物源沙门菌耐药特征分析

为探究不同健康状态和不同饲养环境下沙门菌对临床常用抗菌药物的耐药情况及耐药基因携带情况,采用琼脂稀释法对健康与患病、家养与流浪宠物粪样中分离出的99株沙门菌进行临床常用12种抗菌药物的耐药性检测;用PCR方法对获得的耐药菌株进行相关耐药基因的检测。结果:分离出的宠物源沙门菌对所有被检的抗菌药物耐药率均未超过10.0%,对头孢噻呋、安普霉素、阿米卡星、卡那霉素和硫酸庆大霉素100%敏感;患病动物源沙门菌对被检抗菌药物的耐药率高于健康动物源沙门菌,家养动物源沙门菌对被检抗菌药物的耐药率高于流浪动物源沙门菌(P<0.05)。耐药基因检测结果显示:floR的检出率为3.0%;ant(3″)-Ia、tetA、tetB检出率均为2.0%;bla_TEM、oqxA、aac(6′)-Ib-cr基因检出率均为1.0%,未检出其他被检基因。结果表明,新疆乌鲁木齐市不同健康状态和不同饲养环境下宠物源沙门菌对被检抗菌药物的耐药率及耐药基因携带率低,仅有2株沙门菌存在ant(3″)-Ia+tetA+tetB 3种耐药基因共存情况。根据试验结果发现检出的耐药表型与耐药基因之间存在相关性,...     

2019年湖南省部分地区鸡、鸭、猪源大肠杆菌耐药性调查

旨在调查2019年湖南省部分地区不同来源的大肠杆菌对抗生素的耐药水平,为养殖合理用药提供参考。从4个市畜禽养殖场采集猪、鸡和鸭粪便285份,使用麦康凯琼脂培养基和伊红美蓝培养基对大肠杆菌筛选,利用基质辅助激光解析电离飞行时间质谱进行鉴定,采用微量肉汤稀释法检测16种抗生素对大肠杆菌的最小抑菌浓度(MIC)。共分离出203株大肠杆菌(鸡源146株、鸭源20株和猪源37株),对四环素和氨苄西林耐药率最高,分别达到89.2%和84.7%;对头孢他啶、奥格门丁、黏菌素耐药率较低,分别为2.5%、2.0%和0.5%;所有菌株均对美罗培南敏感。抗3种及以上抗生素的菌株数目占比为87.7%,而仅5.42%的菌株对所有检测的抗生素敏感。另外,来源于减抗示范养殖场的大肠杆菌对大部分抗生素的耐药率,与来源于非减抗养殖场大肠杆菌相比未见显著差异。上述结果表明,湖南省动物源大肠杆菌的耐药水平仍处在较高水平,重视兽药使用管理并持续开展对大肠杆菌的耐药性监测十分必要。     

A survey of antimicrobial resistance in Escherichia coli isolated from wild sika deer (Cervus nippon) in Japan

We examined the antimicrobial susceptibility of 848 Escherichia coli isolates from 237 feces samples of wild sika deer (Cervus nippon) captured between 2016 and 2019 in 39 of the 47 prefectures of Japan. Five of the 237 wild sika deer (2.1%) carried E. coli with resistance to at least one antimicrobial, and all the resistant isolates showed resistance to tetracycline. The resistant isolates contained antimicrobial resistance genes that were similar to those in E. coli derived from humans and farm animals. Although wild sika deer are not currently likely to be a source for the transmission of antimicrobial resistance in Japan, they can potentially mediate antimicrobial resistance spread by coming into contact with humans, animals, and their surroundings.     

抗菌肽的作用机理及其在水产养殖中的应用

本文就抗菌肽的来源和种类、抗菌机理和特点、提高免疫力和抗病力的机理及其在水产养殖中的应用研究进展作一综述。     

Identification and phylogenetic analysis of Babesia parasites in domestic dogs in Nigeria

The present study examined the presence of Babesia parasites in 104 domestic dogs in Nigeria. Sequentially, Babesia parasites infecting domestic dogs underwent genetic and phylogenetic analyses. The results of nested PCR based on the Piroplasmida 18S rRNA gene illustrated that 13.5% (14/104) of the samples were positive. The obtained positive samples determined the nucleotide sequences of the 18S rRNA genes. In the genetic and phylogenetic analyses, four of five nucleotide sequences were similar to Babesia canis rossi, and one sample exhibited a close similarity to a Babesia sp. isolated from a raccoon in Hokkaido, Japan. The present study revealed the widespread presence of B. canis rossi among domestic dogs in Nigeria.     

内蒙古地区奶牛乳房炎链球菌毒力基因检测及耐药性研究

为了解内蒙古地区奶牛乳房炎链球菌毒力基因的分布情况及其耐药现状,通过该地区360份乳房炎奶样链球菌的分离鉴定,利用微量肉汤稀释法测定了81株链球菌对15种药物的MIC,采用PCR法检测耐药基因及毒力基因的携带情况。结果显示,分离到的81株链球菌对β-内酰胺类的耐药率最高;对红霉素和四环素的耐药率在40%~85%;对氟喹诺酮类的耐药率相对较低。多重耐药菌株达88.9%(n=72)。耐药基因pbp2b的检出率为86.4%,tetL和tetM的检出率在75%以上,红霉素耐药表型主要由ermB介导。分离菌株毒力基因中cfb的检出率最高(50.6%),其次为cyl和hylB。内蒙古地区奶牛乳房炎链球菌对β-内酰胺类、大环内酯类和四环素耐药情况严重,且发现多重耐药菌株的流行;链球菌中尤以无乳链球菌的毒力基因携带率较高。     

Blood vitamin concentrations in privately owned dogs fed non-standardized commercial diets and after intake of diets with specified vitamin concentrations

The objective was to investigate in a survey study the blood vitamin concentrations in healthy dogs fed non-specified commercial complete diets and in an intervention study to determine the effects of defined dietary vitamin intakes on blood vitamin levels and hair and skin condition. Sixty-four privately owned dogs, aged from 1 to 8 years, without history of skin or coat problems were included. All animals were fed commercial complete diets with uncertain vitamin concentrations before enrolment. The animals were assigned, according to weight and gender, to four groups with graded vitamin intakes. The blood vitamin levels and skin and coat quality of the dogs were investigated at days 0 and day 122. Coat and hair condition was not influenced by the experimental diets. The retinol concentrations were reduced at the end of the experiment compared with the baseline levels, retinyl esters were not influenced. 25-Hydroxycholecalciferol decreased in all groups, alpha-tocopherol was constant or tended to decrease. Ascorbic acid, thiamine pyrophosphate and riboflavin concentrations were not affected by treatment, flavin adenine dinucleotide and pyridoxal-5'-phosphate were partially reduced on day 122. Cobalamin, pantothenate and biotin concentrations increased with higher dietary intakes, folate levels in tendency. In conclusion, this study gives a survey of blood vitamin concentrations in healthy dogs and provides a data base for the evaluation of the vitamin status in health and disease.     

Analysis of antibiotic resistance phenotypes and genes of Escherichia coli from healthy swine in Guizhou,China

This study was carried out to investigate the resistance phenotypes and resistance genes of Escherichia coli from swine in Guizhou, China. A total of 47 E. coli strains isolated between 2013 and 2018 were tested using the Kirby–Bauer (K–B) method to verify their resistance to 19 common clinical antimicrobials. Five classes consisting of 29 resistance genes were detected using polymerase chain reaction. The status regarding extended-spectrum β-lactamase (ESBL) and the relationship between ESBL CTX-M-type β-lactamase genes and plasmid-mediated quinolone resistance (PMQR) genes were analysed. A total of 46 strains (97.9%) were found to be multidrug resistant. Amongst them, 27 strains (57.4%) were resistant to more than eight antimicrobials, and the maximum number of resistant antimicrobial agents was 16. Twenty antibiotic resistance genes were detected, including six β-lactamase genes blaTEM (74.5%), blaCTX-M-9G (29.8%), blaDHA (17.0%), blaCTX-M-1G (10.6%), blaSHV (8.5%), blaOXA (2.1%), five aminoglycoside-modifying enzyme genes aac(3′)-IV (93.6%), aadA1 (78.7%), aadA2 (76.6%), aac(3′)-II c (55.3%), aac(6′)-Ib (2.1%) and five amphenicol resistance genes floR (70.2%), cmlA (53.2%), cat2 (10.6%), cat1 (6.4%), cmlB (2.1%), three PMQR genes qnrS (55.3%), oqxA (53.2%), qepA (27.7%) and polypeptide resistance gene mcr-1 (40.4%). The detection rate of ESBL-positive strains was 80.9% (38/47) and ESBL TEM-type was the most abundant ESBLs. The percentage of the PMQR gene in blaCTX-M-positive strains was high, and the detection rate of blaCTX-M-9G was the highest in CTX-M type. It is clear that multiple drug resistant E. coli is common in healthy swine in this study. Extended-spectrum β-lactamase is very abundant in the E. coli strains isolated from swine and most of them are multiple compound genotypes.     

饲料中添加天蚕素抗菌肽对南美白对虾免疫功能、抗病力的影响

本研究以南美白对虾为研究对象,探究饲料中天蚕素抗菌肽的合适添加量。在饲料中依次添加不同浓度水平（0%、1%、2%、3%、4%和5%）的天蚕素抗菌肽,配制为6种配合饲料,用以投喂南美白对虾,饲养周期8周。结果显示：与对照组相比,饲料中添加天蚕抗菌肽对南美白对虾增重率、蛋白质沉积率和特定生长率有显著影响（P＜0.05）,高水平天蚕素抗菌肽会抑制南美白对虾的增重率和特定生长率,南美白对虾的饵料系数显著下降,但对南美白对虾存活率和摄食率无显著影响（P＞0.05）|随着饲料中天蚕素抗菌肽的添加水平不断增加,3%组南美白对虾的ACP、POD和SOD含量达到最大水平,与对照组有明显差异（P＜0.05）|随着饲料中天蚕素抗菌肽的添加水平的不断上升,3%组南美白对虾的淀粉酶、脂肪酶和胃蛋白酶含量最高,与对照组存在明显差异（P＜0.05）。结果表明,饲料中添加天蚕素抗菌肽显著提高南美白对虾的增重率和特定生长率,提高南美白对虾的生长发育速度,从而增强机体免疫力,提高南美白对虾的抗病能力。 [关键词]天蚕抗菌肽|南美白对虾|抗病力|免疫功能