罗非鱼嗜水气单胞菌的分离鉴定

１９９８ 年１１ 月份,北京市某温泉鱼场养殖的罗非鱼发生大批死亡,每天死亡达上千斤以上。病死鱼表现：眼眶周边出血,有的眼球突出,背部近鳃盖处出血溃烂,胸鳍基底部出血、糜烂,体表鳞片局部出血。剖检可见胃腺区水肿出血,肠腔积水,重者呈胶冻样,腹腔内有血水,肝胰脏呈土黄色或灰绿色,胆囊肿大,脾脏肿胀并有严重坏死,鳔室内淋巴组织肿大出血。我们从病死鱼及频死鱼肝胰脏中分离到４ 株致病性细菌,４ 株细菌为革兰氏阴性杆菌,无荚膜、无芽胞,有动力,需氧兼性厌氧,经 氧化酶、接触酶、七叶苷水解等生化试验,证明它们是嗜水气单胞菌。４ 株细菌中有３株能与免抗鱼嗜水气单胞菌高免血清发生凝集,１ 株不凝集,这可能是该株细菌具有不同的血清型所致。４ 株细菌均能致死小白鼠及鲢鱼,说明它们为致病性细菌。药敏试验表明,４ 株细菌对环丙沙星、蒽诺沙星高度敏感,而对青霉素和红霉素不敏感。临床应用环丙沙星进行治疗,很快使病情得到控制。     

罗非鱼肠道中弗氏柠檬酸杆菌的分离与鉴定

为了对罗非鱼肠道中弗氏柠檬酸杆菌进行细菌形态学观察、生化试验及药敏试验等研究,试验从罗非鱼肠道中分离、纯化获得2株典型细菌,分别编号为LF-1和LF-2。结果表明:2株细菌均为革兰阴性短杆菌,可以利用枸橼酸盐、葡萄糖产气,其中LF-1鸟氨酸脱羧酶为阴性,而LF-2鸟氨酸脱羧酶为阳性,查询数值编码表确定为弗氏柠檬酸杆菌。药敏试验结果显示,2株细菌均对氯霉素、庆大霉素、头孢孟多、奥复星、诺氟沙星等药物高度敏感。     

罗非鱼套肠病病原菌的分离鉴定及药敏试验

从疑似套肠病的罗非鱼进行细菌分离,人工感染试验确定病原菌后进行药物敏感性试验。结果,从患套肠罗非鱼的肝脏和心脏共分离到2株病原菌LZLF01和LZLF02,API 20NE鉴定结果,2株病原菌均为嗜水气单胞菌。该病原菌对复达欣、先锋霉素VI、菌必治等21种抗生素敏感,对苯唑青霉素和青霉素G耐药。     

25株禽源大肠杆菌的分离和药敏试验

收集了40例具有肝周炎、心包炎及气囊炎的典型病变的病例,无菌操作采集病死鸡和鹅的肝组织接种血平板和麦康凯平板。综合细菌生长和形态学特点及生化试验结果,有25株细菌符合大肠杆菌特点。对25株大肠杆菌进行药敏试验,结果显示：分离菌对痢特灵、菌必治和壮观霉素这3种药物敏感,而对复方新诺明、左氟沙星和多粘菌素B等药物低度敏感或不敏感。由此可见,禽源性大肠杆菌的耐药性强,对该病的治疗增加困难。     

石鲽鱼创伤弧菌病的病原分离鉴定与防治

山东荣成某渔场饲养的石碟鱼大批发病死亡，其体表鳞片局部出血，肝、胰、脾肿胀坏死，腹腔积水等；从病死及濒死石碟鱼的肝、脾中分离到3株细菌，经形态学、生理生化特性及致病因子检测，鉴定为创伤弧菌（Vibrio sulnificus biovar)；3株细菌均不能致死小白鼠。药敏试验表明，3株细菌对环丙沙星、丁胺卡那霉素、卡那霉素、新霉素高度敏感；对氯霉素不敏感。经投喂环丙沙星药饵和腹腔注射卡那霉素，收到良好的防治效果。     

25株禽源大肠杆菌的分离和药敏试验

收集了40例具有肝周炎、心包炎及气囊炎的典型病变的病例，无菌采集病死鸡和鹅的肝组织接种血平板和麦康凯平板。综合细菌生长和形态学特点及生化试验结果，有25株细菌符合大肠杆菌特点。对25株大肠杆菌进行药敏试验，结果显示：分离菌对痢特灵、菌必治、壮观霉素这三种药物敏感，而对复方新诺明、左氟沙星、多黏菌素B等药物低度敏感或不敏感。由此可见，禽源性大肠杆菌的耐药性强，对该病的治疗增加了困难。     

水库网箱养殖吉富罗非鱼暴发死亡的病原菌及其耐药特性研究

为了解水库网箱养殖罗非鱼的死亡病因及病菌的药敏性,对广西大化县岩滩水库库区2个乡镇共5个网箱养殖点吉富罗非鱼暴发死亡的病原菌及其耐药特性进行了研究。按常规方法分离细菌,动物回归试验确定病原菌,以API 20Strep鉴定病原菌,K-B纸片法测定病原菌的耐药特性。调查结果显示,从5个网箱养殖点患病吉富罗非鱼的脑、肝脏和脾脏中共分离到5株病原菌无乳链球菌(Streptococcus agalactiae)。28种药物的药敏试验结果显示,5株病原菌的耐药特性差别较大,都敏感的药物只有舒普深1种,都不敏感的药物有氨苄青霉素、复达欣、复方新诺明等14种。     

养殖型吉富罗非鱼暴发性死亡的病原菌分离鉴定及药敏测定

为查明广西全州县养殖吉富罗非鱼大批死亡的病原及其药物敏感性,为生产中有效防治该病提供参考。按常规方法从患病罗非鱼的腹水、肝脏和心脏中分离细菌,人工感染确定病原菌,以API 20NE生化鉴定和16S rRNA分子鉴定相结合的方法鉴定病原菌并确定其系统发育地位,K-B纸片扩散法检测病原菌的药物敏感性。试验结果,从患病吉富罗非鱼中分离到1株优势菌株GXQZ1,经API 20NE生化鉴定和16S rRNA分子鉴定为类志贺邻单胞菌(Plesiomonas shigelloides),与Plesiomonas shigelloides HRS12816L株的同源性最高(99.3%)。根据人工感染试验结果,认为该菌株是引起广西全州县养殖吉富罗非鱼大批死亡的病原菌。该菌对先锋Ⅴ、先锋Ⅵ、复方新诺明等12种药物高度和极度敏感,对苯唑青霉素等3种药物耐药。     

罗非鱼链球菌的分离鉴定及中草药药敏试验

为了对某养殖户发病的罗非鱼进行治疗,试验进行了病原菌的分离培养、生化鉴定及中药药敏试验。结果表明:从患病罗非鱼体内分离得到一株链状排列的革兰氏阳性球菌,经生理生化鉴定为无乳链球菌,回归试验证明该菌对罗非鱼有较强的致病性。药敏试验结果显示,该菌对苏木、石榴皮、五倍子、黄连、桉树叶、沙棘和乌梅高度敏感,对大黄、黄柏中度敏感。     

野水鸭大肠杆菌病的诊断

广州市某养殖场饲养美国杂交野水鸭暴发以纤维素性心包炎、肝周炎、气囊炎及肠炎为特征的疾病．病鸭除表现精神沉郁及严重下痢外，还表现出头颈歪斜，脚软麻痹，死后角弓反张等明显神经症状。剖检见心包、肝周及气囊不同程度的纤维素性炎，肠粘膜及脑膜充血．可从大多数病死鸭的脑、肝组织均分离出菌落同样的细菌，选取其中６株进行试验，证实为形态、染色及培养、生化特性一致的埃希氏大肠杆菌。皮下注射本菌１８小时向汤培养物０．５ｍｌ／只，能在４日龄樱桃谷鸭复制出本病１０．１ｍｌ／只能致小鼠于１４～４４小时内死亡。本菌经血清学试验鉴定为大肠杆菌０：１血清型。分离获得的６个苗株对盐酸环丙沙星高度敏感；对丁胺卡那霉素、新霉素中度敏感；先锋霉素５低度敏感；对青霉素、链霉素及痢特灵等不敏感。     

肉鸡中嗜水气单胞菌分离与鉴定

从30例临床健康的6周龄肉食鸡泄殖腔内采集棉拭子，接种10日龄SPF鸡胚，发现鸡胚有死亡现象，并从4个样品致死的鸡胚中分离出了细菌。该种细菌的生化培养结果符合法国梅里埃公司细菌鉴定数据库检索系统圳20EV4．0和API 20NE EV6．0所述嗜水气单胞菌(Aeromonas hydrophila)的特征。结果表明临床健康鸡可能携带嗜水气单胞菌。     

北极狐嗜水气单胞菌的分离与鉴定

从北极狐脏器中分离到一种革兰氏阴性杆菌 ,经培养特征、形态、生理生化、致病力测定及药敏试验证实为嗜水气单胞菌 ,确定该菌为引起狐致死的病原菌     

Concurrent infection with ranavirus, Batrachochytrium dendrobatidis, and Aeromonas in a captive anuran colony

Four species (Dendrobates auratus, Phyllobates terribilis, Pyxicephalus adspersus, and Rhacophorus dennysi) of captive anurans with a clinical history of lethargy and inappetence were found dead and were submitted for necropsy. Gross lesions included irregular patches of sloughed skin and rare dermal ulcerations. Histologic findings included epidermal proliferation that was most pronounced on the digits and that included intracytoplasmic chytrid organisms. Bacteria were often associated with the epidermal lesions. Intracytoplasmic inclusion bodies were observed in hepatocytes. Real-time polymerase chain reaction yielded positive results for both Ranavirus and Batrachochytrium dendrobatidis (Bd). Bacterial culture of internal organs yielded Aeromonas hydrophila. This is the first report of concurrent infections in anurans by Ranavirus and Bd and A. hydrophila.     

大型肉鸡养殖场中死淘鸡病原菌分析

本试验通过对山东某地区8个大型集约化肉鸡养殖场的27只死淘鸡进行剖检、电镜观察、病原菌分离鉴定,明确了细菌混合感染为其显著特征,此地区优势致病菌依次为粪肠球菌、浅绿气球菌、大肠杆菌、沙门氏菌、嗜水气单胞菌。用专利产品(专利号:ZL 2009 2 0253251.5)吸管药敏检测盒对死淘鸡病变组织的细菌混合感染进行药敏检测,对照传统的药敏检测结果,表明吸管药敏检测盒具有准确的原位药敏检测的特点和优势,有助于了解肉鸡养殖场潜在的致病因素、提供可行的预防方案。     

Aeromonas hydrophila-associated skin lesions and septicaemia in a Nile crocodile (Crocodylus niloticus)

Aeromonas hydrophila is one of the most common bacteria associated with the aquatic environment. There are, however, limited data on A. hydrophila infection in crocodilians. The aim of this report is to describe a case of skin lesions and septicaemia associated with A. hydrophila in a Nile crocodile (Crocodylus niloticus). A captive male crocodile in the Zoological Park of Antalya (Turkey) was found dead without showing signs of any disease. Gross examination showed brown or red-spotted skin lesions of varying size. These lesions were mostly scattered over the abdomen and occasionally on the tail and feet. At necropsy, numerous white, multifocal and randomly distributed areas were seen on the liver. Gram-stained smears from skin and liver lesions showed Gram-negative bacilli arranged in clusters. Pure cultures of A. hydrophila were recovered from skin, internal organs and blood. Isolates were found to be susceptible to ceftiofur, amoxicillin + clavulanic acid, oxytetracycline, enrofloxacin, danofloxacin, neomycin, gentamicin, and lincomycin + neomycin. A pathogenicity test was performed using this isolate on 4 male 2-year-old New Zealand white rabbits. Local abscesses formed in 2 rabbits injected subcutaneously and the 2 that were injected intraperitoneally died as a result of septicaemia. In conclusion, this report has shown that A. hydrophila may cause skin lesions and even death due to septicaemia in crocodiles.     

疑似草鱼嗜水气单胞菌的分离鉴定及药敏试验

试验对疑似嗜水气单胞菌草鱼的肝脏、脾脏、肾脏组织进行嗜水气单胞菌的分离鉴定。通过普通营养琼脂、胰蛋白大豆琼脂及生化试剂等的分离培养与生化鉴定后,获得3株细菌;16S rRNA通用引物检测3株细菌的基因序列比对确定菌株后,对确定的草鱼嗜水气单胞菌进行回归试验确认其致病性,然后采用临床上常用的阿奇霉素、阿米卡星、链霉素等10种抗生素进行药敏试验。结果表明,经过分离培养、生化鉴定及16S rRNA通用引物鉴定后确定3种菌株分别为嗜水气单胞菌、嗜麦芽窄食单胞菌和腐败斯瓦尼菌,确定的嗜水气单胞菌回归试验证明其具有致病性;药敏试验显示阿奇霉素对腐败斯瓦尼菌和嗜麦芽窄食单胞菌敏感,其抑菌直径分别为105、95 mm,阿米卡星对嗜麦芽窄食单胞菌、嗜水气单胞菌和腐败斯瓦尼菌这3种菌都较敏感,抑菌直径分别为105、95和93 mm。结果表明,疑似为嗜水气单胞菌病的草鱼有3种细菌感染,其中鉴定的嗜水气单胞菌是致病菌,常用抗生素阿米卡星对3种菌都有抑制作用,而仅对腐败斯瓦尼菌和嗜麦芽窄食单胞菌敏感的是阿奇霉素。     

Isolation and Identification of Aeromonas hydrophila from the Case of Channel Catfish Bowel Disease

To ensure the cause of suspected bacteria disease for channel catfish, the bacteria from clinical cases were isolated and cultured, then the biochemical and molecular biology assay, and animal regression test were used to identify the species of the isolated bacteria, bacteria medicine sensitive test was used to detect the bacterial drug resistance. The results showed that the bacteria were gram negative bacilli. The colony morphology and biochemical characteristics of the isolated bacteria were same as Aeromonas hydrophila. The sequencing test of the 16S rDNA also confirmed the isolated bacteria were Aeromonas hydrophila. The isolated bacteria could infect the healthy channel catfish and cause the same symptoms as the clinical case. The isolated bacteria were sensitive to floroquinolones and resistant to penicillin and cephalosporin drugs. The results showed that the pathogen of case catfish bowel disease was Aeromonas hydrophila, and the floroquinolones could use to prevent and treat the disease.     

斑点叉尾鮰套肠病病原嗜水气单胞菌的分离与鉴定

为确定疑似细菌感染的斑点叉尾鮰病原,本试验对临床病例进行细菌分离培养、生物化学鉴定和分子生物学鉴定、动物回归试验和病原菌药敏试验。结果显示,分离菌为革兰氏阴性杆菌,菌落形态和生化特性与嗜水气单胞菌相符;16S rDNA检测与测序分析结果进一步证实分离菌确为嗜水气单胞菌;动物回归试验显示,分离菌能引起与临床自然病例相似的症状;药敏试验显示,分离菌对喹诺酮类药物敏感,对青霉素类、头孢类药物产生耐药性。结果表明,疑似细菌感染斑点叉尾鮰病例为嗜水气单胞菌感染,可选择喹诺酮类药物进行病例的预防治疗。     

鱼嗜水气单胞菌疫苗研究进展

嗜水气单胞菌是一种人兽共患的致病菌,对水产养殖造成巨大危害。己报道的嗜水气单胞菌疫苗有全菌灭活苗、菌体成分亚单位疫苗、减毒活疫苗和DNA疫苗,现分别综述。     

鲤源嗜水气单胞菌分离鉴定、耐药性及毒力基因检测

为查明贵州某鲤鱼养殖厂鲤鱼死亡的病因,本试验进行了细菌分离培养、菌体形态观察、细菌生理生化鉴定、16S rRNA基因测序分析、药敏试验、耐药基因检测、动物感染试验、毒力基因检测等。结果显示,从死亡鲤鱼病变组织中分离到的细菌呈圆形凸起、表面光滑的灰白色菌落,为革兰氏阴性短杆菌;生化鉴定结果显示,分离菌具有运动性,能发酵葡萄糖产酸产气,精氨酸双水解酶、赖氨酸脱羧酶、M-R和V-P试验阳性;应用细菌16S rRNA基因通用引物进行PCR扩增获得大小为1 421 bp的片段,测序发现,该分离菌与16株嗜水气单胞菌相应序列同源性达75.3%~100.0%;药敏试验结果显示,该分离菌对氟苯尼考、头孢曲松、妥布霉素、氧氟沙星、恩诺沙星5种药物呈现高度敏感,对复方新诺明、磺胺异噁唑、羧苄西林等药物呈现耐药,经耐药基因PCR检测显示,该分离菌携带Sul1、Sul2和Intl1 3种耐药基因,与药敏表型相符;人工感染试验显示,该分离菌有致病力,经毒力基因检测显示,该分离菌携带aer、hly和act 3种毒力基因。上述试验结果表明,本研究分离得到了一株耐磺胺类药物的鲤源嗜水气单胞菌,为该鲤鱼养殖厂的鲤鱼疾病防治与合理用药提供了科学依据。