Effect of probiotics, Enteroccus faecium, on tilapia (Oreochromis niloticus) growth performance and immune response

The aim of this study was to analyze the effect of a probiotic bacterium, Enterococcus faecium ZJ4 on growth performances and immune responses of tilapia (Oreochromis niloticus). The tilapias were treated with E. faecium ZJ4 at a final concentration of 1 × 10⁷ cfu ml^− 1 in aquaria water every 4 days. Six aquaria with three replicates for treated and controls were used. After 40 days, the tilapias supplemented with the probiotic showed significantly better final weight and daily weight gain (DWG) than those fed the basal diet (Control) (P < 0.05). There was no remarkable difference (P > 0.05) in the total serum protein, albumin content, globulin concentration and A/G ratio between the treated and control tilapias. The result of lysozyme activity assay was similar to these biochemical indexes. However, the complement component 3 content, myeloperoxidase (MPO) activity and the respiratory burst activity of blood phagocytes were higher (P < 0.05) in E. faecium treated tilapias (trial 1) than the controls.     

嗜水气单胞菌拮抗菌的筛选

用平板划线法或点种法对212株分离自养殖水体和水产动物胃肠道的细菌进行筛选,得到1株对嗜水气单胞菌(Aerom onas hydrophila)有拮抗作用的细菌MLS-7。经过生理生化的初步鉴定,该菌为粪肠球菌(Entero-coccus faecalis)。MLS-7上清液抑菌试验和共培养试验表明其对嗜水气单胞菌有较强的抑菌作用。进一步对MLS-7抑菌谱进行测定,确认其对鱼源的病原菌有一定的抑菌作用。     

致病性粪肠球菌对脊尾白虾(Exopalaemon carinicauda)免疫相关基因表达的影响

为了研究致病性粪肠球菌(Enterococcus faecalis)对脊尾白虾(Exopalaemon carinicauda)免疫相关基因表达的影响,分别对健康脊尾白虾注射生理盐水和粪肠球菌,测定了不同时间点血细胞和肝胰腺中酚氧化酶原(proPO)、C型凝集素(CTL)、超氧化物歧化酶(SOD)、组织蛋白酶D(Cat D)和组织蛋白酶L (Cat L)基因的相对表达量变化.结果显示,与对照组相比,感染粪肠球菌后,血细胞和肝胰腺中proPO基因表达量均于6h达到最大值(P＜0.05),随后逐渐降低至对照组水平;血细胞和肝胰腺中CTL基因表达量分别于12h和6h上升至最大值,随后逐渐降低;血细胞中CTL基因表达量于72 h虽有降低,但仍显著高于对照组(P＜0.05);血细胞和肝胰腺中SOD基因表达量分别于6h和3h达到最大值,随后逐渐降低,并于72 h达到正常水平;血细胞和肝胰腺中Cat D基因表达量变化趋势基本一致,呈现先降低后升高、然后显著降低再升高的趋势;血细胞和肝胰腺中Cat L基因表达量分别于6h和3h显著降低,随后逐渐升高至最大值,并于72 h达到对照组水平.研究表明,长时间的粪肠球菌感染对脊尾白虾免疫相关基因表达影响显著,本研究所用免疫相关基因均可作为粪肠球菌感染疾病的监测指标.     

粪肠球菌累积对虾夷扇贝免疫酶活性的影响

定量分析了粪肠球菌(Enterococcus faecalis)在虾夷扇贝(Pactinopecten yessoensis)中的累积及其组织分布,以及扇贝血淋巴中2种免疫相关酶的活性变化.采用接触染毒法,将扇贝浸泡于含105个细菌细胞/mL的海水中处理2周,采用平板计数法测定了在不同暴露时间和不同扇贝组织中细菌累积数量的变化,并抽取扇贝血淋巴测定了超氧化物歧化酶(SOD)和酸性磷酸酶(ACP)活性.发现处理第7天时,扇贝中含菌量达到最高,为5.20 lg(CFU/g),第14天时又降低到与第3天相当的水平.而累积速率(RA)在第1天时最高,此后均逐渐降低.结果表明,随着在细菌中暴露时间的延长,扇贝累积的细菌数量增长明显减缓.消化道以及外套膜和鳃中细菌含量最高,比软组织平均值高1个数量级以上;而在血淋巴中含量极低,比软组织平均值低2个数量级以上.血淋巴中SOD和ACP酶活性均有先升高后降低的趋势,第7天时2种酶活性均达到最大.实验期间,扇贝血淋巴中免疫酶活性与体内累积的细菌数量变化趋势一致.血清中的可溶性免疫因子对细菌侵染更敏感,2种酶活性与扇贝组织中累积的细菌数量具有明显相关性,但在血细胞中则不明显,由此推测血清可能是扇贝杀灭和消化入侵细菌的茸要场所.     

Comparison of Organic and Inorganic Fertilizers for the Pond Production of Walleye,Stizostedion vitreum

ABSTRACT

Walleye were stocked at 20 fry/m³ and raised for 45-47 days in earthen ponds that were fertilized with organic or inorganic fertilizers. Organically fertilized ponds received periodic doses of alfalfa meal and torrula yeast. Nitrogen and phosphorus levels of 600 ug/L and 30 ug/L, respectively, were maintained by twice-weekly additions of liquid fertilizers in ponds of the inorganic treatment. Growth, survival, and production offish were not affected by the fertilizer treatment. The use of inorganic fertilizers is recommended for the pond production of walleye because it is easier and less costly than traditional methods involving the use of organic fertilizers.     

Dietary supplement of Enterococcus faecalis on digestive enzyme activities,short‐chain fatty acid production,immune system response and disease resistance of Javanese carp (Puntius gonionotus,Bleeker 1850)

This study investigated the effects of Enterococcus faecalis on digestive enzyme activities and short‐chain fatty acid production in fish intestine, resistance against Aeromonas hydrophila and humoral immunity response by 3 experiments on Javanes carp (Puntius gonionotus). The experiment 1 revealed that diet supplemented with E. faecalis significantly (P < 0.05) increased protease and lipase activities compared to control fed fish. Moreover, E. faecalis supplementation significantly enhanced the production of propionic and butyric acid in the intestine, while no significant difference (P > 0.05) in acetic acid production was observed. In the challenge study (experiment 2), fish were injected (intraperitoneal) with 10⁷ A. hydrophila per ml and survival was significantly improved when fish were fed diet supplemented with E. faecalis compared to control fish. In experiment 3, dietary E. faecalis affected immune system response as fish fed the probiont and exposed to 10⁶ A. hydrophila per ml displayed significantly elevated antibody levels compared to control fed fish. Fish fed diet supplemented with E. faecalis but not exposed to the pathogen revealed significantly higher antibody level than control fish (P < 0.05). Therefore, E. faecalis can be used as a probiotic in Javanese carp farming.     

粪肠球菌对吉富罗非鱼的生长、体组成、消化酶活性及血液生理生化指标的影响

为研究饲料中添加不同浓度的粪肠球菌对吉富罗非鱼生长、体组成、消化能力及血液生理生化指标的影响,实验选用初始体质量为(50.59±0.59)g的吉富罗非鱼300尾,随机分成5组,每组设3个重复,每个重复20尾鱼,养殖实验期60 d。分别投喂实测含1.3×10~2(对照组)、1.4×10~5、1.7×10~6、1.5×10~7和1.8×10~8 CFU/g粪肠球菌的5种等氮(36%)等脂(6.75%)的实验饲料。结果显示:(1)罗非鱼的末均重(FBW)、增重率(WGR)、特定生长率(SGR)和尾均摄食量(FI)均在1.5×10~7 CFU/g组达到最大且显著高于对照组,而饲料系数(FCR)显著低于对照组。以FBW、WGR和SGR为评价指标,通过二次回归分析得出,罗非鱼饲料的粪肠球菌适宜添加浓度范围为7.5×10~7~1.1×10~8 CFU/g。(2)各添加组的全鱼粗蛋白含量均显著高于对照组,1.5×10~7 CFU/g组的粗脂肪含量显著高于对照组,各组间全鱼水分和粗灰分含量无显著差异。1.5×10~7 CFU/g组的干物质、蛋白质和脂肪沉积率均达到最大值且显著高于对照组。(3)1.5×10~7和1.8×10~8 CFU/g组的肠脂肪酶活性均显著高于对照组,而1.8×10~8 CFU/g组的肠道蛋白酶活性显著低于对照组;各组间肠道淀粉酶活性无显著差异。(4)1.5×10~7 CFU/g组的平均红细胞体积、血红蛋白浓度和血小板数均显著低于对照组,各组间红细胞数无显著差异。(5)1.8×10~8 CFU/g组的血清中胆固醇、葡萄糖和丙二醛含量显著低于对照组;1.5×10~7 CFU/g组血清中碱性磷酸酶活性显著高于对照组,而谷丙转氨酶和谷草转氨酶活性显著低于对照组。综上所述,吉富罗非鱼(50~210 g)饲料中粪肠球菌的适宜添加浓度范围为7.5×10~7~1.1×10~8 CFU/g,但添加粪肠球菌对血液载氧能力有负面影响。     

Rare serotype occurrence and PFGE genotypic diversity of Streptococcus agalactiae isolated from tilapia in China

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.     

Growth hormone (GH) and reproduction: a review

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.     

Purification and partial characterization of GtHs (cfLH and cfFSH) from Indian walking catfish (Clarias batrachus) (L.) and development of a homologous ELISA for cfLH

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.     

Controlled infection of Poecilia reticulata Peters (guppy) with Tetrahymena by immersion and intraperitoneal injection

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).     

Nitrogenous waste excretion and accumulation of urea and ammonia inChalcalburnus tarichi (Cyprinidae), endemic to the extremely alkaline Lake Van (Eastern Turkey)

The endemic, anadromous cyprinidChalcalburnus tarichi is the only fish species known to occur in alkaline Lake Van (Eastern Anatolia, Turkey). EightC. tarichi were maintained individually in Lake Van water (17 – 19°C; pH 9.8; 153 mEq·I^–1 total alkalinity; 22 total salinity) and tank water samples analyzed for 24 h in 2 to 4 h intervals. At zero time, < 1µM ammonia was present and urea was undetectable in the tank water; at 24 h, total ammonia and urea made up 114±32 and 35±25µM, respectively. Over the experimental period, ammonia-N and urea-N excretion averaged 1041±494 and 607±169moles·kg^–1 fish·h^–1, respectively. The extent of urea excretion was highly variable between specimens. Uric acid excretion was not detectable.Urea was present at high concentrations in all tissues and plasma (25 – 35moles·g^–1·ml^–1) of freshly caughtC. tarichi; total ammonia content of the tissues was by a factor of 1.9 (liver) to 3.0 (brain) lower. High arginase activity (2.4±0.2 U·min^–1·g^–1) was detected in the liver ofC. tarichi but ornithine carbamoylphosphate transferase, a key enzyme of the ornithine-urea-cycle, was absent. Ureagenesis is likely through degradation of arginine and/or uricolysis. High glutamine synthetase activity (11±0.6 U·min^–1·g^–1) and low ammonia content in brain suggest that, like other teleosts,C. tarichi has an efficient ammonia detoxification in the brain, but in no other tissue.Nitrogenous waste excretion at alkaline pH is discussed. The ability ofC. tarichi to excrete high levels of ammonia at extremely alkaline pH is unique among teleosts studied so far. The mechanism of ammonia excretion under Lake Van conditions remains to be elucidated.     

Imported ornamental fish are colonized with antibiotic‐resistant bacteria

There has been growing concern about the overuse of antibiotics in the ornamental fish industry and its possible effect on the increasing drug resistance in both commensal and pathogenic organisms in these fish. The aim of this study was to carry out an assessment of the diversity of bacteria, including pathogens, in ornamental fish species imported into North America and to assess their antibiotic resistance. Kidney samples were collected from 32 freshwater ornamental fish of various species, which arrived to an importing facility in Portland, Oregon from Colombia, Singapore and Florida. Sixty‐four unique bacterial colonies were isolated and identified by PCR using bacterial 16S primers and DNA sequencing. Multiple isolates were identified as bacteria with potential to cause disease in both fish and humans. The antibiotic resistance profile of each isolate was performed for nine different antibiotics. Among them, cefotaxime (16% resistance among isolates) was the antibiotic associated with more activity, while the least active was tetracycline (77% resistant). Knowing information about the diversity of bacteria in imported ornamental fish, as well as the resistance profiles for the bacteria will be useful in more effectively treating clinical infected fish, and also potential zoonoses in the future.     

Effect of iodophor disinfection of non‐hardened Salmo trutta eggs on their bacterial and fungus load

This study investigated the efficiency of iodophor disinfection (135 ppm active iodine for 15–30 min) of non‐hardened Salmo trutta eggs against different groups of bacteria and against fungus. Egg samples were taken from non‐disinfected and from disinfected eggs, microorganisms were cultured on specific nutrient media and their mass was measured by turbidimetric methods. Bacteria and fungus mass of non‐hardened eggs could be reduced but not eliminated by iodophor disinfection with 135 ppm active iodine for 15 min. The extent of reduction was 47–65% (Experiment 1). The efficiency of disinfection increased with disinfection time as the reduction in bacteria and fungus mass was 40–55% after 15 min and 58–74% after 30 min (Experiment 2). Disinfection efficiency of iodophor solution diluted in water (reduction 49–57%) and of iodophor solution diluted in sodium chloride solution iso‐osmolar to the oocytes (reduction 52–61%) was similar (Experiment 3). The reduction in bacteria and fungus mass was persistent as it was 39–72% lower in embryos deriving from disinfected eggs than in embryos deriving from non‐disinfected ones (Experiment 4). In conclusion, the tested disinfection method is inadequate to eliminate pathogens completely but it could positively influence immune defence of eggs and embryos.     

Influence of the antibacterial herbs, Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia on the survival, growth and bacterial load of Penaeus monodon post larvae

The indiscriminate use of antibiotics and chemicals in shrimp hatcheries has led to biomagnification and that in turn could lead to rejection of a whole consignment. The application of the bioencapsulation technique as a tool for curative treatment in shrimp larvae was investigated. Herbs having antibacterial properties such as Solanum trilobatum, Andrographis paniculata and Psoralea corylifolia (methanolic extracts) were bioencapsulated in Artemia and fed to Penaeus monodon post larvae PL 1–25. The post larvae were reared in a medium inoculated with pathogenic bacteria such as Pseudomonas aeruginosa, Staphylococcus aureus, Salmonella typhi and Vibrio sp. Post larvae reared in the non-inoculated water and fed with non-enriched Artemia exhibited 90% survival, highest specific growth rate (12.43%) and reduced bacterial load. P. monodon reared in the bacterial inoculated water and fed with the non-enriched Artemia exhibited the lowest survival (10–30%), specific growth rate (8.42–9.1%) and increased bacterial load (2.86 × 10³ to 3.76 × 10⁵ cfu/g). The methanolic extracts of the herbs helped to increase survival and specific growth rate and reduced bacterial load in the P. monodon culture system. Among the three herbal extracts, P. corylifolia enriched Artemia fed post larvae showed the tendency to higher survival (>50%), growth rate (11.5 averaged) and low bacterial load (1.12 × 10⁵ cfu/g). This revised version was published online in August 2006 with corrections to the Cover Date.     

Status of aquatic and riparian biodiversity in artificial lake ecosystems with and without management for recreational fisheries: Implications for conservation

1. Humanity is facing a biodiversity crisis, with freshwater-associated biodiversity in a particularly dire state. Novel ecosystems created through human use of mineral resources, such as gravel pit lakes, can provide substitute habitats for the conservation of freshwater and riparian biodiversity. Many of these artificial ecosystems are subject to a high intensity of recreational use, however, which may limit their biodiversity potential.
2. The species richness of several taxa (plants, amphibians, dragonflies, damselflies, waterfowl, and songbirds) was assessed and a range of taxonomic biodiversity metrics were compared between gravel pit lakes managed for recreational fisheries (n = 16) and unmanaged reference lakes (n = 10), controlling for non-fishing-related environmental variation.
3. The average species richness of all the taxa examined was similar among lakes in both lake types and no substantial differences in species composition were found when examining the pooled species inventory. Similarly, there were no differences between lake types in the presence of rare species and in the Simpson diversity index across all of the taxa assessed.
4. Variation in species richness among lakes was correlated with woody habitat, lake morphology (surface area and steepness), and land use, but was not correlated with the presence of recreational fisheries. Thus, non-fishing-related environmental variables had stronger effects on local species presence than recreational fisheries management or the presence of recreational anglers.
5. Collectively, no evidence was found that anglers and recreational fisheries management constrain the development of aquatic and riparian biodiversity in gravel pit lakes in the study region; however, the conservation of species diversity in gravel pit lakes could benefit from an increasing reliance on habitat enhancement activities.

     

Epizootics of Streptococcus agalactiae infection in captive rays from Queensland,Australia

The aim of this study was to describe two epizootics of high mortalities from infection with Streptococcus agalactiae, occurring in captive rays held in a marine display aquarium in south‐east Queensland, Australia, in 2009 and 2010. Five different species of rays were affected, including mangrove whiprays (Himantura granulata), estuary rays (Dasyatis fluviorum), eastern shovelnose rays (Aptychotrema rostrata), white‐spotted eagle rays (Aetobatus narinari) and blue‐spotted mask rays (Neotrygon kuhlii). This report describes the history of both epizootics including collection, quarantine and husbandry of rays, the disease epizootics, clinico‐pathological features of the disease, antimicrobial therapy, autogenous vaccine production, and laboratory studies including clinical and histopathology, bacteriology, PCR, molecular serotyping and sequencing of the bacterium S. agalactiae.