微生态制剂对异育银鲫消化酶活性的影响

在基础饲料的基础上，分别添加0.1%、0.2%、0.4%芽孢杆菌，0.3%、0.6%、1.2%硒酵母，混合添加0.2%芽孢杆菌和0.3%、0.6%、1.2%硒酵母，制成9种微生态制剂饲料，以基础饲料为对照，在水泥地中饲养异育银鲫60d。然后，对异育银鲫肠道和肝胰脏的蛋白酶、淀粉酶进行了测定。结果表明，添加0.2%芽孢杆菌，异育银鲫肠道、肝胰脏蛋白酶活性分别比对照组提高466.2%和85.4%，添加0.1%芽孢杆菌，肠道肝胰脏淀粉酶活性分别比对照组提高83.7%和129.5%；添加硒酵母对肠道蛋白酶活性无显著影响，但添加0.6%硒酵母能使肝胰脏蛋白酶性比对照组提高43.7%，添加0.3%硒酵母能使肠道、肝胰脏淀粉酶活性分别比对照组提高84.2%和163.9%；同时添加0.2%芽孢杆菌和0.3%硒酵母能使肠道蛋白酶淀粉酶活性分别比对照组提高395.5%和95.4%，同时添加0.2%芽孢杆菌和0.3%、0.6%、1.2％硒酵母对肠、肝胰脏蛋白酶淀粉酶活性无显著影响。与单独添加0.2%芽孢杆菌相比，同时添加0.2%芽孢杆菌和硒酵母肠道、肝胰脏蛋白酶活性随硒酵母添加量的增加而显著降低，但淀粉酶活性两者之间差异不显著；与单独添加硒酵母相比，0.2%芽孢杆菌和硒酵母同时添加对肠道、肝胰脏蛋白酶、淀粉酶活性促进作用并不显著。     

壳聚糖对异育银鲫的生长、蛋白质合成及肌肉营养成分的影响

采用单因子浓度梯度法 ,将壳聚糖以 1 0 %、 1 5%、 2 0 %的量添加到基础饲料中 ,以基础饲料为对照 ,饲喂异育银鲫 (10 0 60± 10 96g) ,两个月后 ,测定鱼体的相对增重率和饲料系数。结果表明 ,1 0 %组的相对增重率极显著高于对照组 (P <0 0 1) ,比对照组提高 51 44% ,饲料系数比对照组下降2 0 51% ;1 5%组、 2 0 %组的相对增重率显著高于对照组 (P <0 0 5) ,比对照组分别提高 10 44%、13 19% ,饲料系数比对照组分别下降 9 81%、 10 68%。为了进一步了解壳聚糖对异育银鲫的促生长机理 ,测定了白肌的核酸含量和粗蛋白含量。结果显示 ,壳聚糖的添加可以极显著提高白肌的RNA/DNA以及RNA/蛋白质 (P <0 0 1) ,同时 ,白肌的RNA/DNA以及RNA/蛋白质与鱼体的相对增重率存在相关性     

螺旋藻对异育银鲫夏花内源酶、消化率和体成分的影响

将平均体质量3.2 g的异育银鲫夏花随机分为7组,分别在基础日粮中添加0.0%(Ⅰ组)、0.2%(Ⅱ组)、0.5%(Ⅲ组)、1%(Ⅳ组)、1.5%(Ⅴ组)、2%(Ⅵ组)和5%(Ⅶ组)的螺旋藻,饲喂42 d,研究其对异育银鲫肝胰脏和肠道的蛋白酶、淀粉酶活性、蛋白质表观消化率和体成分的影响。结果表明:①螺旋藻对异育银鲫夏花肝胰脏及肠道内的蛋白酶活性有促进作用。Ⅵ组和Ⅶ组肝胰脏蛋白酶活性分别比对照组提高了68.9%和136.47%,差异显著(P<0.05);肠蛋白酶活性分别比对照组提高了11.8%和32.74%,差异显著(P<0.05)。螺旋藻饲料对异育银鲫夏花肝胰脏以及肠道内的淀粉酶影响不明显。②添加螺旋藻的试验组蛋白质表观消化率比对照组高,但差异不显著(P>0.05)。螺旋藻对异育银鲫夏花鱼体的水分、粗蛋白、粗灰分和粗脂肪含量影响不明显;饲喂螺旋藻的6组异育银鲫的粗脂肪含量均有不同程度升高,但与对照组差异不显著(P>0.05)。     

中草药添加剂对异育银鲫生长和蛋白质消化吸收的影响

设计中草药配方Ⅰ和Ⅱ,分别以0.5%、1%、2%剂量添加到基础饲料中,连续投喂异育银鲫2个月后,测定鱼体的相对增重率、肠道和肝胰脏的蛋白酶活性及消化吸收率。结果表明,中草药添加剂能够极显著地影响异育银鲫的生长、肠道和肝胰脏的蛋白酶活性及蛋白质的消化吸收率(P<0.01)。配方Ⅰ以1%组和2%组、配方Ⅱ以2%组的生长最佳,相对增重率分别比对照组提高23.47%、32.74%和47.06%。中草药添加剂对肠道、肝胰脏蛋白酶活性及消化吸收率的影响与生长相似,对照组肠道蛋白酶活性为10.54,配方Ⅰ中1%组为20.33、2%组为22.79,配方Ⅱ中2%组为25.68;肝胰脏蛋白酶活性,对照组为8.54,配方Ⅰ中1%组为14.13、2%配方Ⅰ组为16.59,2%配方Ⅱ组为20.36。对照组的蛋白质消化吸收率为78.15%,1%配方Ⅰ组为84.38%,2%配方Ⅰ组为86.98%,2%配方Ⅱ组为88.19%。因此中草药添加剂提高蛋白酶活力、促进蛋白质的消化吸收,是促进鱼体生长的重要因素。     

乳化剂对异育银鲫消化酶及血清指标的影响

选取8000尾异育银鲫,随机分为2个处理(对照组和试验组),每处理2重复,每重复2000尾,分别饲喂含乳化剂0%和0.05%的基础日粮,试验期为62 d。试验结果表明,饲养31 d后,添加乳化剂使异育银鲫的肝胰脏胰蛋白酶的活性显著提高(P<0.01),肠道胰蛋白酶活性也表现出升高的趋势(P=0.08);血清中总蛋白、球蛋白、葡萄糖和甘油三酯的含量也显著提高(P<0.01)。饲养62 d后,添加乳化剂使异育银鲫肠道淀粉酶活性显著高于对照组(P<0.05);血清中葡萄糖和甘油三酯的含量显著高于对照组(P<0.01)。试验结果表明,饲料中添加乳化剂能够提高异育银鲫的肠道和肝胰脏消化酶的活性,促进脂肪代谢和蛋白质沉积。     

饲料中添加中性蛋白酶对吉富罗非鱼生长和消化功能的影响

在吉富罗非鱼(Oreochromis niloticus)基础饲料中添加中性蛋白酶,添加量分别为0.00%、0.05%、0.1%、0.15%、0.2%和0.25%,研究添加后对吉富罗非鱼生长和消化功能的影响。结果表明,在饲料中添加0.1%~0.25%的中性蛋白酶,吉富罗非鱼的相对增重率和特定生长率显著提高,饲料系数显著降低(P<0.05);添加0.1%~0.2%的中性蛋白酶,显著提高饲料干物质表观消化率和粗蛋白表观消化率(P<0.05);添加量为0.1%~0.15%时,肝胰脏蛋白酶显著高于对照组(P<0.05),添加量为0.15%时,肠道蛋白酶活性显著提高(P<0.05)。综合鱼体相对增重率、特定生长率、饲料系数、消化器官消化酶活性等各项指标,吉富罗非鱼饲料中中性蛋白酶的添加量以饲料的0.1%~0.2%为宜。     

幼鲤肝胰脏、肠道消化酶活性比较研究

采用同日龄幼鲤在同一饲养条件下,连续测得38、76、114日龄时肝胰脏、肠道蛋白酶、脂肪酸和淀粉酶活性。结果表明：肝胰脏、肠道蛋白酶活性,肝胰脏淀粉酶活性随日龄增加而增加;38至76日龄时肠道脂肪酶、淀粉酶活性明显增加,76至114日龄变化不明显,而肝胰脏脂肪酶活性显著增加;38至76日龄时肠道蛋白酶、脂肪酶、淀粉酶活性高于肝胰脏,114日龄时肝胰脏蛋白酶、脂肪酶活性高于肠道,淀粉酶活性与肠道接近。     

淇河鲫鱼消化酶活性研究

淇河鲫鱼肠道中蛋白酶活性显著高于肝胰脏,肠道中蛋白酶活性顺序是前肠<中肠<后肠;肝胰脏和肠道各部分的脂肪酶活性没有显著差异;肠道中淀粉酶活性显著高于肝胰脏,但是肠道中淀粉酶活性前肠与中肠没有显著差异,而后肠达到最高。     

肉碱对草鱼生长性能、体成分和脂肪代谢酶活性的影响

在基础饲料中分别添加100、200、300和400mg/kg DL-肉碱,饲喂平均体重134.3±12.1g的草鱼(Ctenopharyngodon idella)8W,以增重率、饲料系数(FCR),肌肉和肝胰脏中蛋白脂肪含量,肠道脂肪消化酶,肝胰脏脂蛋白脂酶(LPL)、肝脂酶(HL)和脂肪酸合成酶(FAS)等指标为判据,研究肉碱对草鱼生长性能、体成分和脂肪代谢酶活性的影响。结果表明,添加200mg/kg肉碱提高鱼体增重率5.70%(P < 0.05),降低FCR5.26%(P < 0.05);添加200～400mg/kg肉碱显著降低了肌肉和肝胰脏中的脂肪含量(P < 0.05);当肉碱添加量为300mg/kg时,肌肉蛋白含量最高;肉碱对肌肉水分和灰分、肝胰脏的蛋白含量无影响(P > 0.05);添加200、400mg/kg肉碱显著降低了血清胆固醇含量(P < 0.05);在脂肪代谢酶活性方面,200mg/kg肉碱组肠道脂肪消化酶和HL活性较对照组提高28.0%、38.6%(P < 0.05);添加400mg/kg肉碱提高肠道脂肪消化酶、HL及总脂酶活性41.41%、36.26%、10.66%(P < 0.05),降低FAS活性47.3%(P < 0.05)。综上所述,适量肉碱能改善草鱼生长性能、降低肌肉和肝胰脏脂肪含量及促进脂肪降解。建议草鱼饲料中肉碱添加量为200mg/kg。     

低鱼粉饲料中补充蛋氨酸、胆汁酸、牛磺酸对中华绒螯蟹幼蟹生长、饲料利用及抗氧化能力的影响

在低鱼粉(15%鱼粉,LFM)饲料中,分别添加0.5%蛋氨酸、0.5%或1.0%胆汁酸盐、0.5%或1.0%牛磺酸(记为0.5%Met、0.5%Bile、1.0%Bile、0.5%Tau、1.0%Tau)配制5种等氮等能的饲料为5个实验处理组,以全鱼粉组(64.4%鱼粉,FM)和低鱼粉组(15%鱼粉,LFM)为对照,投喂中华绒螯蟹(Eriocheir sinensis)幼蟹[(0.26±0.02)g]8周。结果表明:1)与LFM相比,0.5%Met组增重率和特定生长率显著提高,且各处理组的饲料系数显著降低;FM组蛋白质效率最高,与0.5%Met组、1.0%Bile组、0.5%Tau组、1.0%Tau组均无显著性差异。各组的全蟹体组成差异不明显,仅0.5%Met组的脂肪含量显著高于1.0%Bile和1.0%Tau组。2)LFM组肝胰腺和肠道中的淀粉酶活性最高,FM组最低;肝胰腺总蛋白酶的活性FM组最高,其次是0.5%Met组,两者均显著高于其它各组;FM组和0.5%Met组的肠总蛋白酶活性最高,0.5%Bile组次之;0.5%Met组的肝胰腺和肠道脂肪酶活性最高,1.0%Tau组的肝胰腺、LFM组的肠道脂肪酶活性最低。3)LFM组丙二醛(MDA)含量显著高于FM组和各处理组;FM组的超氧化物歧化酶(SOD)活性和过氧化氢(CAT)酶活性均显著高于其它组,其中LFM组活性最低。结果显示,补充0.5%Met促进了幼蟹的肝胰腺和肠道总蛋白酶和脂肪酶的分泌,显著提高幼蟹的生长和抗氧化性能;添加0.5%、1.0%Bile可有效提高幼蟹肝胰腺、肠道的总蛋白酶和脂肪酶活力,降低饲料系数;而补充0.5%、1.0%Tau均能显著提高幼蟹肝胰腺和肠道消化酶活力和抗氧化能力。补充胆汁酸和牛磺酸未能有效促进生长可能与饲料中的限制性氨基酸不足有关。     

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).     

Evaluation of a Fast Method Based on the Presence of Two Restriction Sites in the Mitochondrial ND5 (mt ND5) Gene for the Identification of Scomber Species

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.     

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.     

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.