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1.
本研究利用MRS培养基,从山东东营刺参(Apostichopusjaponicus)养殖池塘环境中分离到56株乳酸菌。以刺参腐皮综合征2种重要致病菌灿烂弧菌(Vibrio splendidus)和假交替单胞菌(Pseudoalteromonas nigrifaciens)为指示菌,采用牛津杯法测定所分离的乳酸菌对2株致病菌的抑制效果,筛选出1株具有较强抑菌活性的乳酸菌CSND-6,并对该细菌的形态、胞内产物和胞外产物的抑菌活性、生理生化实验、16S r DNA序列分析、菌株生长特性及对刺参安全性进行了研究。结果显示,菌株CSND-6对灿烂弧菌和假交替单胞菌的生长具有较好的抑制作用。其中,胞内产物的抑菌圈分别为17、22 mm,胞外产物的抑菌圈分别为25、31 mm。该菌株对刺参的高浓度浸浴胁迫实验结果表明,所分离的菌株对刺参是安全的。通过生理生化和16S rDNA序列分析表明,CSND-6属于乳杆菌科(Lactobacillaceae),与副干酪乳杆菌(LactobacillusparacaseiJCM1171)的相似性为99.83%。菌株CSND-6在30℃~44℃、pH为6~8范围内生长较快,24 h后进入对数生长期,28~32 h时达到生长高峰,最高值达2.10×109CFU/ml。本研究筛选的副干酪乳杆菌属于刺参池塘养殖环境中土著益生菌,对刺参的主要病原菌抑制效果较好,并表现出了较好的生长特性,可为养殖池塘刺参疾病的生态防控和乳酸菌开发提供参考。 相似文献
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2011年冬季,福建省宁德市霞浦县海区网箱养殖刺参发生"腐皮综合症",并伴有死亡现象出现。刺参的发病症状表现为:厌食、排脏、身体萎缩、体表局部溃烂乃至大面积溃烂。从患病刺参病灶处分离得到两种优势细菌CS1和CS2。经人工回接感染实验证明两种菌对健康刺参都具有较强的感染性,且感染病参的症状与自然发病刺参的症状相同。通过生理生化试验、16S rDNA序列分析及系统进化树分析,结果表明两株菌分别与灿烂弧菌(Vibrio splendidus)和假交替单胞菌(Pseudoalteromonas sp.)相似。菌株CS1与灿烂弧菌的亲缘关系最近,相似率达到99%,菌株CS2与假交替单胞菌的亲缘关系最近,相似率达到95%。菌株CS1可鉴定为灿烂弧菌,菌株CS2可鉴定为假交替单胞菌。另外,在患病刺参呼吸树膜和腔体内发现大量后口虫(Boveria sp.)。所以导致本次刺参"腐皮综合症"的原因可能是致病性细菌和寄生虫共同作用的结果。本文首次揭示了该地区"腐皮综合症"导致养成刺参大规模死亡的致病原因,对刺参病害防治和健康养殖具有重要的指导意义。 相似文献
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2014年12月-2015年12月,在大连地区刺参(Apostichopus japonicus)大水面养殖池塘进行了春、夏、秋、冬四季有益菌分离筛选,从其水体和底泥中共分离得到66株细菌.以刺参“腐皮综合征”主要病原菌——灿烂弧菌(Vibrio splendidus)和假交替单胞菌(Pseudoalteromonas nigrifaciens)为指示菌进行拮抗作用实验,利用选择培养基对菌株产淀粉酶和蛋白酶的能力进行测定,最后通过安全性实验得到潜在益生菌株YQ-2.结果显示,该菌株对灿烂弧菌和假交替单胞菌有较强的抑制作用,抑菌圈分别达到22 mm和24 mm;对淀粉和蛋白选择培养基水解圈的直径达到22 mm和36mm.安全性实验显示,该菌株无论是在108 CFU/ml浸浴还是投喂108 CFU/g的粉末饲料感染,30 d内供试刺参没有发病和死亡现象,健康程度好,且相对于对照组的体重明显增长,108 CFU/g粉末饲料投喂组的相对增长率达到39.31％.此外,本研究对YQ-2菌株的生理生化指标、16S rDNA序列进行了分析,其同源性与枯草芽孢杆菌Bacillus subtilis strain KLP2015相似度达99％,故将该菌株鉴定为枯草芽孢杆菌.该株枯草芽孢杆菌在大水面刺参池塘四季水体中数量为140-280 CFU/ml,高于其他菌株;同时,该菌株在水体中还具有较高的优势度,优势度分别为4.2％、3.5％、2.6％、4.6％,冬、春季节的优势度明显高于夏、秋季节;它属于土著分离菌株,对引起刺参腐皮综合征的2株病原菌具有较强的抑制作用,这对刺参大水面生态养殖具有较大的应用潜力. 相似文献
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为筛选用于微生态制剂研发的海水鱼源益生菌,本研究对海捕野生许氏平鲉(Sebastes schlegelii)和大泷六线鱼(Hexagrammos otakii)的消化道内壁黏膜样品进行细菌分离纯化,获得80株可培养细菌。使用选择性培养基对菌株的产酶能力进行测定,选取海水鱼常见病原菌为指示菌测定分离菌株产物的抑菌活性,筛选出2株潜在益生菌TS2和TH8,并进行菌株的生理生化检测、16S rDNA序列分析、生长特性及其对宿主安全性的研究。结果显示,TS2产蛋白酶(protease)、淀粉酶(amylase)和脂肪酶(lipase),其无菌培养产物可显著抑制鳗弧菌(Vibrio anguillarum)、副溶血弧菌(Vibrio parahaemolyticus)、哈维氏弧菌(Vibrio harvey)和假交替单胞菌(Pseudoalteromonas nigrifaciens)的生长。TH8产蛋白酶和脂肪酶,其无菌培养产物可显著抑制鳗弧菌、溶藻弧菌(Vibrio alginolyticus)、副溶血弧菌、假交替单胞菌、嗜水气单胞菌(Aeromonas hydrophila)、金黄色葡萄球... 相似文献
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刺参养殖池塘一株贝莱斯芽孢杆菌的分离及其生理特性 总被引：1，自引：0，他引：1
2014年5月,在东营的刺参(Apostichopus japonicus)养殖池塘环境中对水样和底泥样品进行了细菌分离和益生菌的筛选,共获得66株可培养细菌。以刺参"腐皮综合征"主要致病菌为指示菌进行拮抗作用实验,使用选择培养基对菌株的产淀粉酶和蛋白酶能力进行测定,筛选获得一株潜在益生菌株——DY-6,并进行了生理生化实验、16S r DNA基因序列分析、菌株生长特性及其对刺参的安全性研究。结果表明,DY-6对其指示菌假交替单胞菌(Pseudoalteromonas nigrifaciens)、灿烂弧菌(Vibrio splendidus)、副溶血弧菌(Vibrio parahaemolyticus)和溶藻弧菌(Vibrio alginolyticus)均有良好的抑制作用,抑菌圈直径分别达到24 mm、22 mm、27 mm和37 mm,对淀粉和蛋白培养基的水解直径分别为28 mm和20 mm。利用该菌对刺参进行高浓度胁迫实验测试其安全性,实验期间浸浴组和投喂组刺参状态良好,没有发病和死亡现象。利用细菌生理生化和16S r DNA基因序列分析表明,DY-6与贝莱斯芽孢杆菌(Bacillus velezensis CP015911)的相似性为99%。因此,初步鉴定该菌株为贝莱斯芽孢杆菌。同时,将该菌株与商品化微生态制剂产品中分离的4株益生菌进行了比较,DY-6在温度20~35℃、盐度0~35范围内生长较快,优于上述4株菌,4 h进入对数生长期,10 h达到生长高峰。筛选的贝莱斯芽孢杆菌具有良好的抑菌能力,生长速度快,并且具有广温广盐的优点,不仅能够降低疾病发生率,还可更好地适应黄河口地区夏季水温偏高、盐度波动大的特点,由此具有良好的益生菌商业化开发潜力。 相似文献
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短小芽孢杆菌X93及胞外产物抑菌活性的研究 总被引：1，自引：1，他引：0
从健康养殖大黄鱼肠道中筛选出对病原弧菌有拮抗作用的潜在益生菌株X93,利用细菌快速鉴定系统和16S rDNA方法进行细菌种类鉴定;通过点种法对其抗菌谱进行测定;采用平板扩散法,研究了培养时间、温度、盐度、蛋白酶K和Fe3+等因素对其胞外产物抑菌活性的影响。结果显示:该菌株为短小芽孢杆菌(GenBank accession No.HM137033);对20株病原指示菌中的13株均产生不同程度的抑制作用,抗菌谱较广;菌株培养48 h后的胞外产物抑菌作用最强;在pH为7时,抑菌活性最大;达到最大抑菌活性的温度和盐度分别为28 ℃、5;蛋白酶K和不同浓度的FeCl3均会降低其抑菌活性。该菌对病原弧菌、迟缓爱德华菌和嗜水气单胞菌等水产上常见主要致病菌有良好的拮抗效果,且具有抑菌作用的胞外产物对温度、盐度和Fe3+有一定的适应范围,适宜作为益生菌株,有望在实际养殖环境中得到应用。 相似文献
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《渔业科学进展》2017,(3)
2014年12月–2015年12月,在大连地区刺参(Apostichopus japonicus)大水面养殖池塘进行了春、夏、秋、冬四季有益菌分离筛选,从其水体和底泥中共分离得到66株细菌。以刺参"腐皮综合征"主要病原菌——灿烂弧菌(Vibrio splendidus)和假交替单胞菌(Pseudoalteromonas nigrifaciens)为指示菌进行拮抗作用实验,利用选择培养基对菌株产淀粉酶和蛋白酶的能力进行测定,最后通过安全性实验得到潜在益生菌株YQ-2。结果显示,该菌株对灿烂弧菌和假交替单胞菌有较强的抑制作用,抑菌圈分别达到22 mm和24 mm;对淀粉和蛋白选择培养基水解圈的直径达到22 mm和36 mm。安全性实验显示,该菌株无论是在108 CFU/ml浸浴还是投喂108 CFU/g的粉末饲料感染,30 d内供试刺参没有发病和死亡现象,健康程度好,且相对于对照组的体重明显增长,108 CFU/g粉末饲料投喂组的相对增长率达到39.31%。此外,本研究对YQ-2菌株的生理生化指标、16S r DNA序列进行了分析,其同源性与枯草芽孢杆菌Bacillus subtilis strain KLP2015相似度达99%,故将该菌株鉴定为枯草芽孢杆菌。该株枯草芽孢杆菌在大水面刺参池塘四季水体中数量为140–280 CFU/ml,高于其他菌株;同时,该菌株在水体中还具有较高的优势度,优势度分别为4.2%、3.5%、2.6%、4.6%,冬、春季节的优势度明显高于夏、秋季节;它属于土著分离菌株,对引起刺参腐皮综合征的2株病原菌具有较强的抑制作用,这对刺参大水面生态养殖具有较大的应用潜力。 相似文献
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带鱼肠道中芽孢杆菌的分离鉴定及其发酵液抗菌性质研究 总被引：1，自引：0，他引：1
从带鱼肠道中分离筛选到1株具有广谱抗菌活性的菌株JFL15,其发酵液对嗜水气单胞菌、美人鱼发光杆菌、哈维氏弧菌、溶藻弧菌、大肠杆菌、迟钝爱德华氏菌、铜绿假单胞菌、副溶血弧菌、创伤弧菌、查氏弧菌均有较强抑菌作用。通过形态特征、生理生化特征、16S rDNA序列分析以及系统发育进化分析,菌株JFL15与暹罗芽孢杆菌的同源性最高,达98.05%,初步鉴定为暹罗芽孢杆菌。对暹罗芽孢杆菌JFL15的发酵液进行研究发现,暹罗芽孢杆菌JFL15的发酵液对酶、温度、酸碱度和紫外线均不敏感,且发酵产物储存较长时间后仍具有较强抑菌活性。 相似文献
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从对虾养殖池中分离出1株编号为2013082515(简称菌株15)的菌株,以鳗弧菌(Vibrio anguillarum)、哈维氏弧菌(Vibrio harveyi)和副溶血弧菌(Vibrio parahaemolyticus)为指示菌,分析了菌株15对指示菌的抑菌效果及最低抑菌浓度,同时,分析了菌株15对其他7株弧菌的抑菌效果.结果显示,菌株15对3株指示菌均具有抑菌效果,对鳗弧菌、哈维氏弧菌及副溶血弧菌的最低抑菌浓度分别为2.50×104、2.50×105、2.50×105 CFU/ml;对其他弧菌也具有一定的抑菌效果.采用注射及浸泡感染方法分析了菌株15对对虾的生物安全性,结果显示,在高浓度时,菌株15对对虾具有潜在毒性.分别用细菌全细胞脂肪酸气相色谱法和16S rDNA序列分析比对法对该菌进行分类鉴定,表明菌株15为一株假交替单胞菌(Pseudoalteromonas sp.). 相似文献
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用改良二倍稀释法和抑菌圈法测定了24种单方中草药和9种复方中草药对刺参腐皮综合征4种重要病原菌灿烂弧菌、假交替单胞菌、哈维氏弧菌和恶臭假单胞菌的体外抑菌活性。结果显示,不同中草药对特定病原菌抑菌效果差异较大,单方中草药最低抑菌浓度（MIC）为1.56mg/ml,中草药水煎剂H-5和H-9对假交替单胞菌的抑菌圈最大,达22mm。单方中草药中穿心莲、大青叶、金银花和川芎4种中草药对4株病原菌抑菌效果最好,其水煎剂对4株病原菌平均抑菌浓度分别为6.25、7.81、5.86和3.52mg/ml;平均抑菌圈直径分别是17.5、11.3、14.0和11.5mm。将筛选的4种单方中草药组成9种复方,复方MIC和抑菌圈均显著好于单方,最低MIC降低到0.20mg/ml。其中复方HC-G和HC-D抑菌浓度最低,效果最好,平均抑菌浓度分别降低为0.54和0.64mg/ml;平均抑菌圈直径分别为15.3mm和16.3mm。试验结果显示,复方最佳配比为穿心莲、大青叶、金银花和川芎=2∶1∶3∶2。该复方可为生产高效专用中草药,以替代抗生素,为刺参健康养殖提供技术保障。 相似文献
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Association between the interannual variation in the oceanic environment and catch rates of bigeye tuna (Thunnus obesus) in the Atlantic Ocean 
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下载免费PDF全文 The environmental processes associated with variability in the catch rates of bigeye tuna in the Atlantic Ocean are largely unexplored. This study used generalized additive models (GAMs) fitted to Taiwanese longline fishery data from 1990 to 2009 and investigated the association between environmental variables and catch rates to identify the processes influencing bigeye tuna distribution in the Atlantic Ocean. The present findings reveal that the year (temporal factor), latitude and longitude (spatial factors), and major regular longline target species of albacore catches are significant for the standardization of bigeye tuna catch rates in the Atlantic Ocean. The standardized catch rates and distribution of bigeye tuna were found to be related to environmental and climatic variation. The model selection processes showed that the selected GAMs explained 70% of the cumulative deviance in the entire Atlantic Ocean. Regarding environmental factors, the depth of the 20 degree isotherm (D20) substantially contributed to the explained deviance; other important factors were sea surface temperature (SST) and sea surface height deviation (SSHD). The potential fishing grounds were observed with SSTs of 22–28°C, a D20 shallower than 150 m and negative SSHDs in the Atlantic Ocean. The higher predicted catch rates were increased in the positive northern tropical Atlantic and negative North Atlantic Oscillation events with a higher SST and shallow D20, suggesting that climatic oscillations affect the population abundance and distribution of bigeye tuna. 相似文献
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Liping Li Rui Wang Wanwen Liang Xi Gan Ting Huang Yan Huang Jian Li Yunliang Shi Honglin Luo Ming Chen 《广西水产科技》2014,(1)
Previously, we reported 10 PEGE types of 85 tilapia Streptococcus agalactiae(GBS), which shifted from Streptococcus iniae in China, by using PEGE method. Presently, larger and more representative tilapia GBS were isolated, for the ?rst time in China, to characterize their serotypes and genetic diversities more precisely than had done before. 168 GBS strains were distributed in ?ve provinces of China, in which Guangdong, Guangxi and Hainan were the major ones, holding36.9%(62/168), 37.5%(63/168) and 19.6%(33/168), respectively. Serotypes, Ia, Ib and III, were observed in these strains and the most predominant one was Ia(95.2%), which mainly distributed in Guangdong, Guangxi and Hainan. Ia initially occurred in 2009, it shoot up to 32.1% in 2010,but decreased to 16.1% in 2011 before went up to 45.2% in 2012. Ib sporadically occurred during2007–2011, III onlyoccurred in 2012. 14 different PFGE types, including 4 new types(N, O,P and Q), were observed, in which B, D, F and G were the predominant types, holding 83.9%(141/168) of the total GBS strains. Ia corresponded to 11 PFGE types(A–H, N–P), in which type D predominated(51%). Ib represented 3 genotypes(I, J and Q) and III harbored only 2genotypes(N and F). Type N and Fsynchronously presented in Ia and III. In summary, the genetic diversity of tilapia GBS varied by serotypes and changed with geographical locations and years.Although Iastillpredominated, new rareserotypeIII alreadyoccurred in China. 相似文献
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Iciar Martinez Amaya Velasco Ricardo Pérez-Martín Eskil Forås Michiaki Yamashita Carmen G. Sotelo 《Journal Of Aquatic Food Product Technology》2013,22(4):289-297
The purpose of this work was to evaluate the suitability of a method based on the presence of two restriction sites (for Hae III and Hindf I) in the mitochondrial NADH dehydrogenase subunit 5 (mt ND5) gene to identify Scomber species. The evaluation was performed on 144 reference and market samples by sequencing of the entire 505-bp fragment of the mt ND5 gene and of a 464-bp fragment of the Kocher fragment of the cytochrome b gene (mt Cytb). Sequence analysis of any of the two fragments allows the identification of each of the four Scomber species, but S. japonicus and S. colias had the same restriction sites at the ND5 amplicon and would not have been differentiated by this analysis. Similarly, loss of the Hae III site in some S. scombrus individuals would have misidentified them as not being Scomber. All the market products were correctly labeled except one acquired in Spain labeled as originating in the Atlantic and containing S. japonicus. 相似文献
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In this experiment, a feeding trial was performed to determine the effects of fructooligosaccharide (FOS) on growth performance, digestive enzyme activity and immune response of Japanese sea bass, Lateolabrax japonicus juveniles (initial weight 38.3 ± 0.5 g), and the fish were examined following feeding with six levels of FOS (0, 0.5, 1, 2, 4 and 6 g/kg) for 28 days. Significant enhancement of weight gain (WG) and specific growth rate (SGR) was found in fish fed 1 g/kg FOS incorporated diets (p < .05), while the feed conversion ratio (FCR) in the 1, 2 g/kg FOS groups reduced significantly compared with the control (p < .05). Besides, the crude lipid in the 4, 6 g/kg FOS groups increased significantly compared with the control (p < .05). On the other hand, the erepsin and lipase activities significantly elevated in intestine of fish fed 2 g/kg FOS (p < .05) and the lysozyme activity in serum of fish fed 2 g/kg FOS were significantly higher than that in the control (p < .05). Moreover, the alkaline phosphatase activities in serum of fish fed 0.5, 1, 2 g/kg FOS were significantly higher than in control (p < .05). Regression analysis showed that the relationships between dietary FOS levels and either SGR, FCR, erepsin or lysozyme activities were best expressed by regression equations, and the optimal inclusion levels are 1.37, 1.80, 3.06, 3.11, 1.93 and 1.80 g/kg for SGR, FCR, erepsin, lipase, lysozyme and total superoxide dismutase activities, respectively. Overall, this study revealed that FOS incorporated diets could beneficial for L. japonicus culture in terms of increasing the growth, digestion and immune activities. Under the present experimental condition, the optimal supplementary level of FOS in the diet of L. japonicus is 1–3 g/kg. 相似文献
17.
Growth hormone (GH) and reproduction: a review 总被引：5，自引：0，他引：5
Florence Le Gac Odile Blaise Alex Fostier Pierre-Yves Le Bail Maurice Loir Brigitte Mourot Claudine Weil 《Fish physiology and biochemistry》1993,11(1-6):219-232
Interaction between growth and reproduction occurs in many vertebrates and is particularly obvious at certain stages of the life cycle in fish. Endocrine interactions between the gonadotropic axis and the somatotropic axis are described, the potential role of GH being emphasised. A comparative analysis of these phenomena in mammals, amphibians and fish, suggests a specific role of GH in the physiology of puberty, gametogenesis and fertility. It also shows the original contribution made by studies on the fish model in this field of investigations. 相似文献
18.
Two gonadotropins (GtH; Qa and Qb) were purified by gel filtration and ion exchange chromatography from the pituitaries of Indian walking catfish (Clarias batrachus). The presence of GtH during purification was assessed by in vitro oocyte maturation and in vivo steroidogenic activity, and their identities were determined by elution profiles, molecular weight, biological activities and yield. The molecular weights of Qa and Qb were 37 and 42 kDa, respectively, and composed of distinct subunits (Qa: 20 and 14 kDa and Qb: 26 and 18 kDa). Polyclonal antibodies raised against Qa immunostained Qa, Qb and pituitary GtH cells. A competitive Qa‐ELISA was developed whose sensitivity was 6.25 ng mL?1 (1.25 ng well?1) with intra‐ (3.5%) and inter‐ (12.4%) assay coefficients of variation. Displacement curves parallel to the standard were obtained with plasma and pituitary extracts of catfish, Qb and carp GtHII. The assay was validated by measuring the plasma Qa levels after LHRH treatment and in relation to ovarian growth in the female catfish during different reproductive phases. Based on the results, Qa and Qb corresponded to fish LH and FSH respectively. The findings will increase the knowledge of the mechanisms controlling fish reproduction and identification of sensitive phases in fish in captivity for hormonal manipulation. 相似文献
19.
Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection 下载免费PDF全文
下载免费PDF全文 G Sharon M Pimenta‐Leibowitz M C L Vilchis N Isakov D Zilberg 《Journal of fish diseases》2015,38(1):67-74
Tetrahymena is a protozoan parasite, which infects guppy, Poecilia reticulata Peters, and causes substantial economical losses in commercial farms worldwide. Studies of guppy infected by Tetrahymena require standardized infection protocols. The LD50 for Tetrahymena infection of guppies by intraperitoneal (IP) injection was calibrated, and the level obtained was 946 parasites per fish. Guppy infection with Tetrahymena by immersion, imitating the natural route of infection via the integument, was studied under normal or stress conditions. Exposure to cold and netting (CNI) and to cold only (CI) followed by immersion exposure to 10 000 Tetrahymena per mL resulted in 22.5% and 19.2% mortality, respectively, as compared to 14.2% and 10% in groups that were netted only (NI) or non‐stressed (I). Histopathology revealed that immersion infection resulted in a systemic infection. Lysozyme levels, measured 3 weeks after infection, were significantly higher in the CNI group (288 μg per mg protein) compared with CI‐, NI‐ and I‐treated groups (94.5, 64 and 62.3 μg mg?1, respectively). There was no evident parasite immobilization activity in body homogenates, suggesting no development of acquired immunity. Re‐infection by IP injection revealed no increase in protection in any of the treatment groups, mortality range of 56.3–75%, higher than in the non‐exposed control (40.6% mortality). 相似文献
20.