黄鳝温室炼苗期致病性类志贺邻单胞菌的分离与鉴定

对湖北仙桃某养殖场温室黄鳝(Monopterus albus)炼苗不同时间段的患病黄鳝进行病原分离,从7日龄、14日龄和21日龄苗中分离得到1株优势菌株,命名为B-1,对分离菌株B-1进行药敏试验和动物回归试验;同时对养殖池塘的水质指标进行测定,并对患病黄鳝肝、脾组织进行了病理学研究。药敏试验结果表明,分离菌株B-1对氟苯尼考、盐酸多西环素、硫酸新霉素、氯霉素和环丙沙星敏感,对恩诺沙星、红霉素、庆大霉素、阿奇霉素耐药。经16S rRNA基因序列测定分析、生化检测及动物回归试验,确定B-1菌株为病原菌——类志贺邻单胞菌(Plesiomonas shigelloide)。水质各项指标检测结果显示,水体环境有一定的恶化。组织病理学观察显示,不同时期黄鳝苗的肝、脾组织都发生了较明显的病变。     

患病大鲵类志贺邻单胞菌的分离与鉴定

从患病濒死大鲵(Andrias davidianus)肝脏中分离到1株病原菌YZ01,经Biolog微生物系统鉴定及16S rDNA序列的系统发育分析,均表明该病原菌为类志贺邻单胞菌(Plesiomonas shigelloides)。人工感染试验可致健康大鲵出现与自然患病大鲵相同的症状,并从感染患病大鲵体内再次分离到该病原菌。药物敏感试验表明,该菌株对头孢曲松、头孢噻肟、氯霉素和头孢呋辛等18种药物敏感。     

异育银鲫类志贺邻单胞菌的鉴定

采用API 20E系列生化鉴定、生理生化特征、电镜观察和16S rDNA序列构建系统发育树分析方法,对从濒死的异育银鲫(Carassius auratus gibelio♀×Cyprinus carpio♂)肝脏中分离到的1株致病菌(革兰氏阴性杆菌,YCS07-01株)进行了鉴定。结果显示:菌株主要生化特性为氧化酶阳性,发酵葡萄糖产酸不产气,分解肌醇;不发酵蔗糖、甘露醇、鼠李糖等;β-半乳糖苷酶、精氨酸双水解酶、赖氨酸脱羧酶、鸟氨酸脱羧酶阳性。负染电镜观察,菌株为两端钝圆直杆状,无芽胞,无荚膜,有4根鞭毛,大小为0.8~1.0μm×3.0μm。在以该菌16SrDNA序列(GenBank登录号FJ375179)和GenBank数据库内同源性较高的细菌16S rDNA序列构建的系统发育树中,分离菌YCS07-01与类志贺邻单胞菌ATCC14029T(登录号X74688)的同源性为99%,并聚为一个分支。综合培养特征、形态和生理生化特性、系统发育分析等将该菌鉴定为类志贺邻单胞菌(Plesiomus shigelloides)。药敏试验结果表明菌株对羧苄青霉素和万古霉素耐药;对头孢唑啉、四环素和氨曲南中度敏感;对丁胺卡那霉素、左氟沙星、头孢他啶、复方新诺明等高度敏感。     

鳜致病性类志贺邻单胞菌的分离鉴定及耐药性分析

2019年江苏扬州某养殖场鳜出现大量死亡,临床症状表现为体表溃疡、鳃丝腐烂等,解剖发现肝脏出血。为查明鳜死亡病因,自濒死鳜分离到优势生长细菌,通过形态观察、理化特性测定及16S rRNA基因同源性分析进行分离菌的种属鉴定,同时通过人工感染试验、毒力基因检测确定其致病性,并通过纸片扩散法进行耐药性分析。结果表明：引起鳜大量死亡的病原菌株SN1为类志贺邻单胞菌,其对鳜的半数致死密度为1.05×10⁵ cfu/mL;类志贺邻单胞菌SN1携带astA、astB、astD、astE、actP、aphA等6个毒力相关基因;类志贺邻单胞菌SN1对青霉素G和克林霉素2种药物耐药,对哌拉西林、苯唑西林和麦迪霉素等6种药物中介,对恩诺沙星、硫酸新霉素、盐酸多西环素、氟苯尼考等26种药物敏感。本试验结果可为类志贺邻单胞菌引起鳜细菌性疾病的防治提供理论依据。     

斑点叉尾鮰致病性类志贺邻单胞菌的分离鉴定及药物敏感性

为探究某斑点叉尾鮰养殖场鱼类发病死亡原因,从患病的斑点叉尾鮰(Ietalurus Punetaus)鱼体分离得到一种病原菌,通过形态学观察以及16S rRNA基因序列测定与系统发育树构建分析,确定该菌为类志贺邻单胞菌(Plesiomonas shigelloides)。用Kirby-Bauer纸片扩散法检测病原菌的敏感性,进行药敏试验。类志贺邻单胞菌为革兰氏阴性菌,且对呋喃唑酮、头孢拉定、头孢曲松、头孢唑林及头孢呋辛5种药物产生高度敏感,对氧氟沙星、头孢氨苄、哌拉西林等11种药物中度敏感。通过回归感染试验进行验证,注射类志贺邻单胞菌菌株后,斑点叉尾鮰感染后出现的症状与自然发病个体的症状基本相同。从患病鱼体分离出的即为类志贺邻单胞菌,也正是该致病菌导致斑点叉尾鮰死亡,可利用头孢类高度敏感药物防治。     

类志贺邻单胞菌耐药基因检测与耐药性关系的研究

为研究类志贺邻单胞菌的耐药表型和耐药基因之间的关系,采用K-B纸片扩散法对55株类志贺邻单胞菌进行耐药性鉴定,同时采用PCR方法对相关耐药基因进行检测。药敏试验结果显示,55株类志贺邻单胞菌对30种抗生素药物产生不同程度的耐药性,其中对克林霉素(98.2%)、万古霉素(98.2%)、氨苄西林(90.9%)、麦迪霉素(89.1%)、羧苄西林(89.1%)、苯唑西林(87.3%)的耐药性较高。TEM、gyrB和tetR基因PCR检出率分别为18.2%、67.3%和36.4%,其中39株致病菌至少含有1个耐药基因。试验结果表明,分离所得的全部类志贺邻单胞菌均多重耐药,耐药基因和耐药表型之间呈现一定的相关性,但非一一对应。     

大口黑鲈源维氏气单胞菌的分离鉴定

为查明2020年春季广东省佛山市某养殖场人工养殖大口黑鲈(Micropterus salmoides)鱼苗出现暴发性死亡的病因,该研究对患病大口黑鲈鱼苗肝脏组织中分离的一株细菌GZXR2020进行了分子鉴定,PCR扩增获得该菌株的16S rRNA基因和gyrB基因全长序列,BLAST显示其与维氏气单胞菌(Aeromon...     

类志贺邻单胞菌间接ELISA检测方法的建立

自患病草鱼分离的类志贺邻单胞菌菌株LCCiL90625NA经甲醛灭活后注射新西兰大白兔,制备兔抗血清,Protein A亲和层析柱分离纯化抗体,通过优化反应条件,建立类志贺邻单胞菌间接ELISA检测方法.结果表明,制备的兔抗类志贺邻单胞菌多克隆抗体效价为1∶1.28×105,建立的ELISA检测方法可特异性地检测类志贺邻单胞菌,纯化后的多克隆抗体与副溶血弧菌、创伤弧菌、溶藻弧菌、非O-1霍乱弧菌、嗜水气单胞菌、豚鼠气单胞菌、温和气单胞菌、易损气单胞菌和杀鲑气单胞菌等细菌均无交叉反应,检测灵敏度为1.0×104 CFU·mL-1.该方法的建立为类志贺邻单胞菌的快速检测和该病的早期诊断及防控提供基础.     

大口黑鲈病毒性溃疡病病原的分离和鉴定

对广东省佛山地区2008年夏季高温时期暴发的大口黑鲈溃疡病的病原进行分离,从患病鱼的病灶肌肉组织中分离到5株嗜水气单胞菌,人工感染健康大口黑鲈,均未出现溃疡暴发病的典型症状病鱼体表大片溃烂,裸露肌肉坏死并有出血,尾鳍、胸鳍和背鳍基部红肿溃烂。制备病灶肌肉组织的除菌过滤液,背部肌肉注射感染健康大口黑鲈,7 d后出现典型的溃疡病症状。取自然发病鱼和人工感染的患病鱼病灶肌肉组织制作超薄切片,经电镜观察,均发现组织中有大量病毒颗粒,病毒粒子有囊膜,呈六角形,为正20面体对称结构,大小约为145.5 nm。根据已知虹彩病毒主要衣壳蛋白（MCP）基因序列设计特异引物,提取人工感染发病鱼病灶肌肉组织的DNA进行PCR扩增,将扩增片段进行序列测定与分析,结果表明该序列与已报道的虹彩病毒MCP基因有着较高同源性（39.5％～100％）。从电镜观察和MCP基因测序分析结果确认该病毒为虹彩病毒科蛙病毒属中的一种病毒,与国外报道的大口黑鲈病毒（LMBV）在分子特性和引起的疾病特征上有一定差异。研究结果表明引起大口黑鲈溃疡性暴发病的病原是虹彩病毒,将该病暂命名为大口黑鲈病毒性溃疡病。     

Temporal genetic structure of an intergrade largemouth bass (Micropterus salmoides) population

Since 1972, numerous impoundments throughout Texas have been stocked with Florida largemouth bass Micropterus salmoides floridanus. In order to examine some of the long-term effects of such efforts, the present study summarizes past studies and complements those with more current observations on the intergrade largemouth bass population in Aquilla Lake, Hill Country, Texas. The reservoir received 4 annual stockings during the month of May from 1982 to 1985. Collections occurred periodically from February 1984 to June 1987, February to August 1988, and monthly from August 1991 to November 1992. Horizontal starch-gel electrophoresis was conducted to determine allele frequencies at loci sAAT-B*, SOD*1. Mean heterozygosity and percent introgression were highest for age-0 fish of the 1986 year class. Assortative mating was not apparent in 1991 or 1992 year classes. Collections of specific year classes over time consistently indicated decreases in the relative abundance of the northern phenotype. Relative to the northern phenotype, the abundance of Florida year classes remained relatively constant over time. Relative abundances of F₁ hybrids and F_x, intergrades (i. e., F₁) from specific year glasses were observed to generally increase with age.     

Isolation and identification of a viral haemorrhagic septicaemia virus (VHSV) isolate from wild largemouth bass Micropterus salmoides in China

Fish rhabdoviruses are a family of viruses responsible for large‐scale fish die‐offs worldwide. Here, we reported the isolation and identification of a member of rhabdoviruses from wild largemouth bass (Micropterus salmoides) in the coastal area of the Pearl River Estuary, China. This virus isolate was identified as viral haemorrhagic septicaemia virus (VHSV) by specific RT‐PCR. Furthermore, the virus (VHSVLB2018) was isolated by cell culture using fathead minnow cells and confirmed by RT‐PCR. Electron microscopy showed the presence of bullet‐shaped viral particles in the cytoplasm of infected cells. The complete sequencing of VHSVLB2018 confirmed that it was genome configuration typical of rhabdoviruses. Phylogenetic analysis based on whole‐genome sequences and G gene nucleotides sequences revealed that VHSVLB2018 was assigned to VHSV genogroup Ⅳa. The pathogenicity of VHSVLB2018 was determined in infection experiments using specific pathogen‐free largemouth bass juveniles. VHSVLB2018‐infected fish showed typical clinical signs of VHSV disease, including darkened skin, petechial haemorrhages and pale enlarged livers, with the cumulative mortalities reached 63.3%–93.3% by 7 days post‐infection. VHSVLB2018 was re‐isolated from dead fish and confirmed by RT‐PCR. Together, this is the first report of isolation and identification of a VHSV isolate from wild largemouth bass in China.     

Persistence of Florida largemouth bass alleles in a northern Arkansas population of largemouth bass, Micropterus salmoides Lacèpéde

Abstract– Lake Ashbaugh, located in northeast Arkansas, was constructed in 1981, and initially stocked with Florida largemouth bass followed by supplemental stockings of northern largemouth bass. Allele frequencies of three discriminant allozyme loci (sAAT-B, sIDH-B, sMDH-B) between Florida and northern largemouth bass were determined for 414 largemouth bass collected between 1994 and 1996. F_x bass dominated our sample, with 62.3% possessing Florida largemouth bass alleles. A high incidence of Hardy-Weinberg disequilibrium was observed, indicative of genetic change within the population. No significant differences were identified for frequency of age classes, relative weight, and length at age between the northern, F₁ and F_x phenotypes. Despite being located north of what is generally considered suitable for stocking Florida largemouth bass, it was demonstrated that temperature is not selective at present against bass possessing Florida largemouth bass alleles. However, caution should prevail when introducing non-native stock into native gene complexes, as introduced genes persist through many generations.     

Evaluation of alternative pond production systems for raising largemouth bass,Micropterus salmoides

This study evaluated three different pond‐based production systems for raising largemouth bass, Micropterus salmoides, for the food fish market, using nine 0.04‐ha ponds. Treatments included traditional ponds (TP), intensively aerated ponds (IAP), and split‐pond systems (SPS). TP and SPS ponds were aerated at 9.3 kW/ha, while IAP was aerated at 18.6 kW/ha. TP was stocked at 7,500 fish per ha (three replicates per treatment), and the other two production systems (SPS, IAP) were stocked with 12,500 fish per ha. Feed‐habituated advanced fingerlings (128 ± 47.6 g mean individual weight) were cultured for 157 days. Fish were fed a formulated diet (42% protein, 16% lipid) four times a day, feeding with a maximum allowance of 3% of total body weight and readjusted to the initial body weight biweekly. Fish raised in the SPS displayed a significantly lower specific growth rate, lower individual final weight, and lower weight gain, but the biomass gained was significantly higher than TP but not IAP. Final biomass gained was 50% higher in the SPS and IAP than in the TP. Survival rate and feed conversion ratio were not significantly different among treatments and ranged from 71 to 79% and 1.64 to 2.14, respectively.     

Effects of season, maturity, and sex on lipid class dynamics in largemouth bass (Micropterus salmoides Lacepede)

Abstract Factors that can influence changes in lipid class concentrations in specific tissues have not been reported for largemouth bass Micropterus salmoides. We investigated patterns of lipid composition by measuring seasonal levels of total lipid and major lipid classes in the muscle and liver tissues of juvenile and adult largemouth bass and gonad tissue of adults. Fish were collected in December, February, April and July to correspond with key stages of the sexual cycle. Total lipid concentrations were substantially greater in hepatic (32.8 [female] to 56.6 [male] mg. g^?1) and gonadal tissues (34.4 [F] to 204.0 [M] mg. g^?1) than in muscle (7. 5 [F] to 11.4 [M] mg. g^?1). Muscle lipid concentrations in juveniles were significantly higher (10.9 [F] and 11.4 [M] mg. g^?1) in summer than in winter in adult muscle tissue (8.6 [F] and 9.4 [MI mg. g^?1). A 44% increase in ovary lipid was observed from preparatory to prespawn stages, followed by a 78% decline after the spawn. Additionally, seasonal hepatic lipid concentrations were substantially higher in juveniles. Concentrations of lipid classes (free cholesterol [C], triacylglycerol [TAG], free-fatty acids [FFA], esterified cholesterol [CE] and phospholipid [PL]) of all tissue types varied significantly among season, sex, and maturity categories. Changes with all lipid classes in the ovary coincided with those occurring in total ovarian lipid. Mobilization and transport of TAG and PL from the liver to the ovary appeared to occur during gametogenesis. All class concentrations in juveniles were invariably higher in liver than in muscle, but seasonally variable among hepatic and gonad tissues of adults. Juvenile males showed slightly greater hepatic C concentrations than other groups. Muscle and hepatic C were generally higher in juveniles.     

Diet of the largemouth bass, Micropterus salmoides (Lacepède), in Lake Naivasha, Kenya

Abstract The major food items of adult largemouth bass, Micropterus salmoides (Lacepède), in Lake Naivasha, Kenya are not markedly different from those in its native range. Although insects and their larvae are major components of the diet for both temperate and equatorial populations of juvenile bass, the equatorial population also eats Procambarus clarkii (Girard), juvenile fish and aquatic weeds. In temperate populations they also eat crustaceans, rotifers and oligochaetes. The frequency of occurrence of the major prey organisms in the stomachs of M. salmoides varies seasonally with population peaks in these organisms. Similar to North American populations, the Naivasha population feeds during the day between dawn and dusk. In contrast to temperate populations, the population of M. salmoides in this equatorial lake feeds throughout the year, with feeding intensity correlated with water temperature.     

饥饿对大口黑鲈消化器官、蛋白酶和淀粉酶活力的影响

对饥饿和饥饿后投喂的大口黑鲈(Micropterus salmoides)蛋白酶和淀粉酶活力及消化组织的变化进行了研究,饥饿周期为30d。饥饿对大口黑鲈消化道指数的影响为肝体比从试验的第3天开始明显下降(P<0.05),第20天开始保持稳定水平(P>0.05);幽门盲囊重与体重比先上升后下降;胃重与体重比始终呈上升趋势(P<0.05);肠重与体重比在第10天开始保持稳定水平(P>0.05);肠长与体长比呈上升趋势(P<0.05)。饥饿对蛋白酶活力影响为从开始到第3天胃蛋白酶活力上升,第10天开始下降;幽门盲囊蛋白酶活力在第10天之后持续下降(P<0.05);肠道蛋白酶活力一直呈下降趋势。饥饿对淀粉酶活力影响为肝脏淀粉酶活力、胃淀粉酶活力和幽门盲囊淀粉酶活力均呈下降趋势(P<0.05);肠道淀粉酶活力第10天之后即稳定在低水平(P>0.05)。饥饿后摄食对蛋白酶活力和淀粉酶活力的影响为饥饿后摄食蛋白酶活力和淀粉酶活力均呈上升趋势,上升的速度在不同组织有差异,在恢复摄食后的第10天基本达到正常水平。     

Dietary synbiotics improved the growth,feed utilization and intestinal structure of largemouth bass (Micropterus salmoides) juvenile

This study investigated the effect of dietary synbiotics on the growth, feed utilization and intestinal histology of largemouth bass (Micropterus salmoides). A commercial synbiotic product containing dried powder of yeast, Bacillus subtilis and mannan oligosaccharides was supplemented in basal diet with 0 g/kg (control), 1.0, 2.0, 4.0 and 6.0 g/kg, respectively, and then, the five diets were fed to largemouth bass with body weight of 4.5 ± 0.1 g for 8 weeks. The results showed that the supplementation of 2.0, 4.0 and 6.0 g/kg synbiotics increased weight gain by 20.6%, 17.7% and 11.8% (p < .05) and decreased feed conversion ratio by 0.20, 0.18 and 0.12 (p < .05), respectively, when compared to the control. The protein retention, lipid retention, apparent digestibility coefficient of dry matter and crude protein, and the intestinal villus height and width of 2.0 and 4.0 g/kg synbiotic groups were significantly higher than those of the control (p < .05). The intestinal protease activity and serum lysozyme activity were increased by the addition of 2.0 and 4.0 g/kg synbiotics (p < .05). In conclusion, dietary synbiotics improved the growth, feed utilization and intestinal structure of juvenile largemouth bass, and the recommended inclusion level was 2.0–4.0 g/kg.