稻田养殖的大鳞副泥鳅出血病病原菌的分离鉴定与药敏试验

为确定稻田养殖的大鳞副泥鳅(Paramisgurnus dabryanus)出血病病原与防治措施,应用微生物分离技术和分子生物学相关方法,对患出血病的大鳞副泥鳅进行病原菌的分离和鉴定并分析其药物敏感性。从患出血病大鳞副泥鳅中分离到3株优势菌(命名为DLFNQ-1、DLFNQ-2、DLFNQ-3);回归感染实验结果显示,仅DLFNQ-1对大鳞副泥鳅具有致病性,半致死浓度为2.3×10~7 CFU/mL;生理生化试验表明,DLFNQ-1菌株对葡萄糖、阿拉伯糖、氧化酶、七叶灵等反应为阳性,对柠檬酸、尿素酶等反应为阴性,与嗜水气单胞菌的生理生化特性相符;DLFNQ-1菌株的16S rDNA序列与嗜水气单胞菌(JN391411.1)的相似度达到99.7%,N-J进化树显示其16S rDNA与其它嗜水气单胞菌的聚为一支。DLFNQ-1菌株对庆大霉素、妥布霉素这两种抗生素高度敏感,对阿洛西林、哌拉西林、卡那霉素等中度敏感,对青霉素、阿莫西林、四环素等呈现出一定的耐药性。结果表明,引起稻田养殖大鳞副泥鳅出血病的病原为嗜水气单胞菌(Aeromonas hydrophila),稻田养殖时可选用庆大霉素、妥布霉素进行防控。     

大口黑鲈源鳗弧菌的分离鉴定

为探明引起大口黑鲈(Micropterus salmoides)体表及内脏出血病的病原，本研究从发病大口黑鲈肝、脾、肾处分离病原，得到1株形态一致的优势菌株2004301,采用腹腔注射方式进行人工回感实验，证实菌株2004301能够引起养殖大口黑鲈内脏出血症状并死亡，且具有较强毒力。综合形态学观察、生理生化特性分析、基于16S rRNA基因和hsp 60基因构建的系统发育树及特异性基因检测结果，将菌株2004301鉴定为鳗弧菌(Vibrio anguillarum)。菌株2004301对32种药物的敏感实验证实其对恩诺沙星、氟苯尼考、磺胺甲恶唑、环丙沙星、氧氟沙星等抗菌药物敏感。     

下口鲇烂尾病病原的分离与鉴定

对患烂尾病下口鲶(Hypostomus plecostomus)的肝脏、脾脏、肾脏进行病原菌分离,分离到NY635菌株,用NY635菌株人工感染体重在20～30 g的健康鱼,人工感染的下口鲇出现与自然发病鱼相似的症状,并从人工感染的下口鲇病灶中分离出菌落形态相同的细菌。初步证明NY635是引起下口鲇烂尾病的病原。对NY635进行菌落形态观察、生理生化鉴定和16S rDNA基因序列分析,确定菌株NY635为碱性假单胞菌(Pseudomonas alcaligenes),并对其进行了药敏试验。     

罗非鱼源蜡样芽孢杆菌分离、鉴定及药敏特性研究

从福建漳州某养殖场患出血病死亡的罗非鱼肝脏、肾脏及脾脏分离到一株致病性菌株FJLF。对该菌进行了形态特征观察、理化特性测定及16S rRNA序列分子鉴定。生理生化特征和16S rRNA基因序列分析结果显示,分离到的FJLF株为蜡样芽孢杆菌(Bacillus cereus)。人工感染该菌后发病鱼出现与自然发病类似症状,且从病灶中分离到与原感染菌一致菌株。腹腔注射后该菌株对罗非鱼的半致死剂量LD_(50)=2.30×10~6CFU/g。FJLF株对诺氟沙星、头孢噻肟、新霉素及卡那霉素等8种抗生素高度敏感;对环丙沙星、痢特灵、红霉素及链霉素中度敏感;对苯唑西林、青霉素、头孢拉定及阿莫西林等10种抗生素耐药。     

中华鳖源粘质沙雷氏菌分离、鉴定及药敏特性研究

对患病中华鳖(Trionyn sinensis)进行病原分离、鉴定及药敏实验。从患病中华鳖肝、肾、脾及腹水分离纯化病原菌,经理化特性测定及16S rRNA序列分析对其进行鉴定,开展人工感染试验,并利用K-B进行药敏特性分析。结果显示分离菌株HD01为本次引发中华鳖病害的病原菌,其对中华鳖的LD50为4.48×106CFU/g。HD01株理化特性与粘质沙雷氏菌一致,16S rRNA序列与粘质沙雷氏菌同源性为99%,综合判定分离菌株为粘质沙雷氏菌(Serratia marcescens)。HD01株对氟苯尼考、多西环素、庆大霉素及苯唑西林等14种抗生素高度敏感;对青霉素、头孢拉定、新霉素等7种抗生素耐药。分离菌株HD01是中华鳖病原菌,养殖时可选用氟苯尼考、庆大霉素及多西环素等药物进行防控。     

黄鳝源嗜水气单胞菌DB01分离鉴定及药敏特性研究

从患病的黄鳝(Monopterus albus)体内分离到一株致病菌株DB01,对该菌株进行人工感染、生理生化鉴定及16S r DNA序列分析。结果显示:人工感染黄鳝出现与自然发病相同的症状,表明菌株DB01为发病黄鳝的病原菌。菌株DB01为革兰氏阴性杆菌,无芽孢和荚膜,在兔血平板上能形成明显的β-溶血。序列分析显示,菌株DB01与嗜水气单胞菌(Aeromonas hydrophila)的同源性达99%。药敏试验结果显示,菌株DB01对诺氟沙星、左氧氟沙星、阿奇霉素及氟苯尼考等抗生素高度敏感,五倍子、黄柏、地榆及乌梅等中草药对菌株DB01抑菌作用强。     

一例甲鱼穿孔病病原的分离鉴定和药敏试验

为研究甲鱼穿孔病的有效预防措施,对患病甲鱼进行了临床诊断和病原分离鉴定。病原菌鉴定(菌落特征、细菌形态及细菌生化和分子鉴定)以及菌株毒性试验、人工感染试验和药物敏感试验的结果表明:致病菌株为嗜水气单胞菌,菌株有明显的β溶血,对小鼠具有很强的致病性,菌液浸泡感染黄鳝,也能使黄鳝出现出血病症状。该菌对嗯诺沙星和四环素高度敏感,对氨苄青霉素和阿莫西林低度敏感并有耐药性。     

养殖牙鲆致病性弧菌的鉴定及药敏分析

2020年丹东东港市某养殖厂养殖1龄牙鲆(Paralichthys olivaceus)突发病害,表现为腹水病症状。取患病的5尾养殖牙鲆的鳃、肝、脾、肠道、腹水为材料进行细菌分离及纯培养,选择优势菌株通过全自动细菌分析仪及16S rDNA序列分析对该菌株进行鉴定,结果表明分离纯培养的14株菌(DS200401-DS200414)共鉴定出2种,其中1种为该病的致病菌:弧菌属灿烂弧菌(Vibrio splendidus)。随后进行人工感染试验和药敏试验。用12种抗菌药物对3种菌进行药敏试验,结果显示在不同菌株间无明显的敏感与耐药性差异。     

革胡子鲶出血性败血症病原菌的分离鉴定

从暴发性死亡的革胡子鲶(Clarias gariepinus)中分离出可疑病原菌株2010111403(简称1403),分别采用细菌全细胞脂肪酸鉴定系统、Biolog微生物自动鉴定系统及细菌16 S rDNA序列分析三种方法对其鉴定,结果表明菌株1403为嗜水气单胞菌(Aeromonas hydrophila)。分离菌株对健康革胡子鲶致病性测试表明其对革胡子鲶的半数致死剂量(LD50)为6.32×106菌落形成单位(CFU);实验感染革胡子鲶出现与自然发病相似症状,表明菌株1403是引起革胡子鲶发生暴发性死亡的主要致病病原。药敏试验表明,庆大霉素、四环素、阿奇霉素、氟哌酸、先锋霉素等9种抗生素对分离菌株有较强的抑制作用,但分离菌株对复方新诺明、克林霉素、罗红霉素等4种药物表现出耐药性。     

天峡红鮰出血病的病原及其防治研究

从患出血病的天峡红蛔进行了病原菌的分离得出2株细菌,经过人工感染实验确立H-I菌株为致病菌株,并对该菌形态培养特性,生理生化鉴定,确定该菌为豚鼠产气单胞菌.药敏试验该菌对呋哺妥因,氯霉素、丁胺卡那最为敏感,经过生产防治试验取得了较好的效果.     

2种体色黄鳝性别、体长及体重之间的关联分析

为了解不同体色黄鳝的性别与体长、体重的差异,2011年5-7月,于洞庭湖地区随机采集灰色及黄色2种体色共208尾野生黄鳝,并对体长、体重及性别进行测量鉴定。样本中,雌性146尾,雄性62尾,雌雄比例为2.35∶1。其中黄色黄鳝83尾,灰色黄鳝125尾。分析结果表明,黄色黄鳝雌雄个体体重及肥满度均高于灰色群体;黄色黄鳝雌性体长大于灰色雌性,雄性小于灰色雄性。雌雄个体体重与体长的相关系数,黄色黄鳝分别为0.914、0.945,灰色黄鳝分别为0.567、0.438。黄色雌鳝比例随着体长的增加逐渐降低,雄性比例随体长的增加逐渐升高,雄鳝在35 cm以上占优势,40 cm之上雄性仅1尾。灰色雌鳝在40 cm以下占优势。2种体色黄鳝雌性比例与体重变化无明显规律。     

Effects of Estradiol Valerate on Steroid Hormones and Sex Reversal of Female Rice Field Eel,Monopterus albus (Zuiew)

The rice field eel, Monopterus albus (Zuiew), is an important aquaculture fish in many Asian countries. Productivity, however, is limited by low fecundity as a result of sex reversal of female juveniles of this fish. This study was conducted to investigate the effects of estradiol valerate on the sex reversal process of this fish by determining sex changes of initial 2‐yr‐old female eels (26.5 ± 2.6 cm, 21.9 ± 2.1 g) treated with three concentrations of estradiol valerate: 0 mg/kg (Control, Group A), 10 mg/kg (Group B), and 50 mg/kg (Group C). In Group A, sex reversal took place with significantly higher percentage of sex change: about 45% of female eels to intersexes and males. Only about 22% of female eels changed to intersexes and males in Group B, but no sex reversal occurred in Group C during the experimental period. The result also indicated that the gonadal development and the concentrations of serum estradiol (E₂) and testosterone (T) varied significantly with different concentrations of dietary estradiol valerate. Serum T significantly reduced but serum E₂ significantly increased in Group C compared to the control Group A. The results demonstrated that estradiol valerate affected the secretion of sex steroid hormones in rice field eels and thus inhibited the sex reversal process at a low concentration (10 mg/kg) or stopped the sex reversal process at a high concentration (50 mg/kg) of estradiol valerate. These results suggest that a higher yield of mature female eels could be achieved by treatments with estradiol valerate, thus enhancing the fecundity of this rice field eel.     

Human Vibrio vulnificus infections and environmental isolates in the Netherlands

Abstract A 63-year-old man was admitted to hospital for septicaemia and severe metastatic skin infection, 24h after he had eviscerated fresh eels. V. vulnificus was isolated from his blood and wounds. The strain was indole negative, was ornithine decarboxylase positive and grew at 42°C. A strain of V. vulnificus with these characteristics was isolated in 1987 from diseased eels. The characteristics differed from those of V. vulnificus strains (biogroup 1) that have been reported from patients world-wide. V. vulnificus biogroup 1 was isolated from 3 of 11 seawater samples collected along the coast of the Netherlands, but indole negative strains of V. vulnificus were not isolated. We conclude that an indole-negative variant of V. vulnificus is pathogenic for eels and for human beings and that eels may transmit V. vulnificus to humans.     

Anosmic Japanese eel <Emphasis Type="Italic">Anguilla japonica</Emphasis> can no longer detect magnetic fields

ABSTRACT:   Magnetosensitivity in anosmic Japanese eel Anguilla japonica was examined by conditioning and electrocardiography. Five eels from a freshwater farm in Kagoshima, Japan, were made anosmic by injection of boiling petroleum jelly (130–140°C) into the nasal cavity, a technique widely used in studies on olfaction and olfaction-related behavior of eels. Another five eels were not injected with boiling petroleum jelly and served as controls. Conditioning consisted of exposure of eels to an imposed magnetic field of 192 473 nT. After complete conditioning of the eels, the imposed magnetic field was reduced from 192 473 nT to 12 663 nT during the tests. The control eels showed a significant conditioned response (i.e. slowing of the heart beat) to the magnetic field after one set of conditioning runs. The anosmic eels did not respond to the magnetic field, presumably due to damage of the olfactory organ from the jelly injection. The results suggest that Japanese eels have a magnetosensitive organ in or around the nares. At 1 month after the anosmia treatment, there was no visible trauma to the olfactory lamellae of anosmic eels.     

Effects of dietary lipids on tissue fatty acids profile,growth and reproductive performance of female rice field eel (<Emphasis Type="Italic">Monopterus albus</Emphasis>)

The effects of different lipids on tissue fatty acid profile and reproductive performance in female rice field eel were investigated in this study. Virgin female eels were fed with six diets containing different lipids (diets FO, LO, SO, PO and PL with fish oil, linseed oil, soybean oil, peanut oil and pork lard, respectively; diet APO with arachidonic acid and peanut oil). The results showed that there were positive correlations between the contents of 18:2n-6, 18:3n-3, arachidonic acid (ARA), eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) in the tissues of eels and those of the corresponding fatty acids in their diets. The specific growth rate of eels fed with diet PO was the lowest and significantly lower than that of FO and SO. Gonad of eels fed with diets PO and PL showed hypogonadism. The long chain polyunsaturated fatty acids (LC-PUFA) can be synthesized by eels, but the quantity was not enough to meet their reproduction requirement completely. The fatty acid desaturation, rather than elongation probably was one of the limiting factors. Addition of proper amount of ARA in diet was favorable to the increase of the hatching rate of fertilized eggs, while EPA and DHA in diet were beneficial to the increase of the survival rate of larva. Both n-3PUFA and a suitable n-6/n-3PUFA ratio were necessary for growth and reproduction of eels.     

Evaluation and characterization of a novel probiotic Lactobacillus pentosus PL11 isolated from Japanese eel (Anguilla japonica) for its use in aquaculture

In the present study, a potential Lactobacilli probiotics were isolated from Japanese eels (Anguilla japonica) and characterized and evaluated for their possible use in eel farming. Sixteen Lactobacilli were isolated from intestines of Japanese eels, using selective media. The lactobacilli strains (represented as PL1 to PL16) were screened by their ability to produce digestive enzyme. Among these, three strains (PL11, PL13 and PL16) producing four digestive enzymes (amylase, cellulase, protease and phytase) simultaneously were characterized further using API ZYM kit. From these, PL11 (Lactobacillu (L.) pentosus) was identified as potential probiotics candidate producing 15 enzymes among 20 tested. Further examination of biological activities of PL11 revealed tolerance against pH, artificial bile juice and antibacterial activity against several fish pathogenic bacteria. The in vitro competitive exclusion assay also revealed 88.4% reduction in adhesion of fish pathogen (Edwardsiella tarda) by PL11 to host intestinal mucus. In vitro incubation of Japanese eel foregut with Baclight‐labelled PL11 showed colonization of the enterocyte surface by confocal and scanning electron microscopy. In summary, PL11 isolated from eels could serve as a potential probiotics with acid and bile tolerance, production of digestive enzymes, antibacterial activity and inhibition of fish pathogen adhesion to intestinal mucus.     

Influence of geomagnetism on the activity and orientation of the eel, Anguilla anguilla (L.), as evident from laboratory experiments

Abstract– Studies were performed with both a mechanical and a photosensory arrangement of the experimental tanks. Silver (migratory) eels were more active than yellow (stationary) eels. Comparison of pooled data on yellow eels tested under natural geomagnetic field conditions with those on eel tested under unnatural ones prcded no evidence of difference in activity. Silver and yellow eels showed stronger activity during the 6 days around new moon compared with the phase around full moon as well as a peak before new moon and a smaller peak before full moon. Under each kind of unnatural field condition, the activity relationships reversed or were cancelled. The directional choice of eels under natural conditions (controls) pointed to magnetic NW and mostly also in the opposite direction. When the magnetic north was changed by 90°, the tested yellow eels followed this change to about the same degree. The additional change of both the inclination (from 68° or 45° or 30°) and total intensity (by a reduction to 31% of the natural field) resulted in a change of the preferences of about 90° relative to magnetic north. Reversal of magnetic north by 180°, which is identical to a change of the inclination from 68° to 112°, also resulted in a change of the preferences by about 90°. Three experiments using compensation of both the horizontal and the vertical geomagnetic field led to circular directional preferences at random in two cases and in one case to about NW of the residual field magnetic north. The navigational abilities of the eel, on the basis of its magnetic sensitivity, are discussed.     

Vibrio vulnificus serovar A: an emerging pathogen in European anguilliculture

The spread of the emerging pathogen Vibrio vulnificus biotype 2 serovar A in Danish anguilliculture is reported. Serovar A was originally isolated in a Spanish eel farm in 2000 and occurred in Denmark in the summer of 2004, affecting eels of 5-10 g body weight cultured in fresh water. The Danish eels showed clinical signs different from those reported for Spanish eels, such as severe haemorrhages in the head and gill region with necrosis of the soft tissues. Danish isolates were biochemically and serologically identical to Spanish serovar A strains and also highly virulent for eels by both intraperitoneal injection and immersion challenges. Vaccination with Vulnivaccine, a vaccine against V. vulnificus serovar E, cross-protected eels against serovar A. The LD(50) for experimentally infected vaccinated animals was significantly higher than for non-vaccinated animals.     

Notes on the occurrence of two tropical species of Anguilla in reservoirs in south-eastern Sri Lanka and preliminary data on the populations

Abstract. The occurrence and abundance of eels were investigated in five lowland reservoirs in south-eastern Sri Lanka. Paired fyke nets and longlines were used for sampling eels. Two tropical species of eel. Anguilla bicolor McClelland and Anguilla nebulosa McClelland, were caught. In two of the five reservoirs investigated only A. bicolor was found and in another two reservoirs both species were caught. Eels seemed to be absent from one reservoir. The CPUE in reservoirs with eel was between 0·13 and 2·60 eels of both species per fyke net and night, and the total weight was between 81 and 814g. A small capture-recapture experiment in Badagiriya, where eels seemed to be most plenliful, gave a population estimate of only six eels or 1·2 kg per ha (eels longer than 40cm). Such a low abundance was regarded as unrealistic. Stocks of eel could presumably, at least in some reservoirs, withstand commercial exploitation but more information is needed on recruitment, growth and migration before any advice can be given on the management of stocks.     

Evaluation of head-only and head-to-tail electrical stunning of farmed eels (Anguilla anguilla, L.) for the development of a humane slaughter method

The overall objective was to evaluate the suitability of electronarcosis as a stunning method for farmed eels. In the first experiment the minimum electrical current needed to induce a general epileptiform insult by head‐only stunning was assessed. The individual eels (n = 40) with a live weight of 700–800 g were fixed in a specially designed re‐strainer. The EEG (electroencephalogram) and ECG (electrocardiogram) recordings, observation of behaviour and responses to pain stimuli were used to assess unconsciousness, insensibility and cardiac function. The applied current of 150, 200 or 250 V, 50 Hz, AC was delivered via scissor‐model stunning tongs for approximately 1 s. A general epileptiform insult was observed in 31 eels for which a successful EEG recording was obtained, using 255 ± 4 V, 545 ± 32 mA, for 1.2 ± 0.2 s. The general epileptiform insult as measured on the EEG was characterized by a tonic/clonic phase and an exhaustion phase. The behaviour showed one phase: tonic cramps alternated by clonic ones. The heart rate was 22 ± 8 beats min^?1 (n = 23) prior to stunning. After stunning the ECG revealed fibrillation. In the second experiment the behaviour of seven individual eels able to move freely in water was observed after head‐only stunning (250 V). Two phases were distinguished. Limited tonic and clonic cramps combined with backward swimming were followed by heavy clonic cramps combined with unco‐ordinated movements such as jumping out of the water. A distinct exhaustion phase was not observed in all animals. In the third experiment a head‐to‐tail electrical method was examined in 15 eels for rendering the eels unconscious and insensitive prior to slaughter. They were stunned by applying 253 V for 3 s followed by 50 V for 5 min. In the fourth experiment nine eels were head‐only stunned with 260 V for 1 s immediately followed by 50 V for 5 min applied from head to tail. Results obtained in these two experiments were similar. After stunning no brain activity and no responses to pain stimuli on the EEG were observed and the ECG showed ventricular extra systolae. It was observed that it might take 60 ± 25 min or longer for a complete recovery. It can be concluded that for effective electrical stunning of eels with a weight of 700–800 g an average current of 545 ± 32 mA (at approximate 250 V, 50 Hz AC) is needed. In this case, within a confidence level of 95% at least 91% of the eels are effectively stunned (n = 31). Therefore, it is recommended to increase the minimum current for an effective stun in practice to 600 mA. Further research is needed to determine the conditions to induce permanent unconsciousness and insensibility of the eels to protect the animals at slaughter.