斑点叉尾鲴嗜麦芽寡养单胞菌的分离鉴定及系统发育分析

从发生在四川、重庆等省市的斑点叉尾妇急性流行性传染病病鱼的肝脏、肾脏内分离到1株高致病性菌株（CCF00024），人工感染健康鱼表现出与自然痛鱼相同的症状，并从中分离获得同种细菌，证实其为斑点叉尾鲴急性流行性传染病的病原菌。形态、生理生化检测表明，该菌为非发酵型直杆菌，严格需氧，革兰氏阴性，极生多鞭毛，对除麦芽糖和甘露糖以外的多种糖类不利用产酸，氧化酶阴性，DNA酶、蛋白酶、脲酶、赖氨酸脱羧酶阳性，MR、VP阴性。在以该菌16SrDNA序列（GenBank登录号AY970826）和GenBank及RDP数据库内同源性较高的细菌16SrDNA序列构建的系统发育树中，分离菌CCF00024与嗜麦芽寡养单胞菌（Stenotrophomonasmaltophilia）聚在一簇，特剐是与5．maltophiliaM5—1的同源性最高，序列相似性达99．6％，结合形态和生理生化特点将其鉴定为嗜麦芽寡养单胞菌（Stenotrophomonasmaltophilia）。药敏试验结果表明，对磺胺甲口恶唑、磺胺异口恶唑、阿齐霉素、洛美沙星高度敏感，而对新霉素、卡那霉素、氨苄青霉素、头孢唑啉、先锋霉素V和链霉素不敏感。     

斑点叉尾鮰对3种致病菌灭活菌苗的免疫应答

将经福尔马林灭活的柱状嗜纤维菌(Cytophaga columnaris)、嗜水气单胞菌(Aeromonas hydrophi-la)和叉尾鮰爱德华菌(Edwardsiella ictaluri)作为免疫原,分别接种斑点叉尾鮰(Ictalurus punctatusRafinsque)后,通过测定受免鱼的交叉凝集抗体效价、头肾和血液中吞噬细胞的吞噬活性及用A.hydrophila活菌攻毒的方法,探讨了斑点叉尾鮰对3种灭活菌苗的免疫应答状况。试验结果表明,对斑点叉尾鮰经腹腔注射接种3种灭活菌苗能刺激受免鱼产生较强的免疫应答,受免鱼产生了对3种致病菌的特异性凝集抗体和交叉凝集抗体,与未接种菌苗的对照鱼相比,受免鱼头肾和血液中吞噬细胞的吞噬活性明显上升,活菌攻毒的结果也证明了受免斑点叉尾鮰对C.columnaris,A.hydrophila和E.ictaluri产生了不同程度的相对免疫保护率(RPS)。     

Effects of Partial and Complete Replacement of Synthetic Cryoprotectant with Carrot (Daucus carota) Concentrated Protein on Stability of Frozen Surimi

ABSTRACT

The study aimed to evaluate the effects of carrot concentrated protein (CCP) as additive on the functional and textural properties of surimi from striped catfish (Pangasianodon hypophthalmus) during six months of frozen storage (?20°C). The CCP (82.22% crude protein) was used as an additive either a lone or with a synthetic cryoprotectant (sucrose-sorbitol-sodium tri-polyphosphate). Control was made with synthetic cryoprotectant only. Molecular weight of CCP was found to be 36 kDa. After six months, the results revealed that up to 50% of synthetic cryoprotectants could be replaced by CCP during frozen storage of surimi. Biochemical parameters such as protein solubility, Ca²⁺ATPase activity, and gel strength decreased significantly (p < .05) during storage. Treatment T-3 (CCP 0.5% + 50% of synthetic cryoprotectant) maintained quality of protein significantly superior (p < .05) in respect to denaturation and other functional and sensory attributes compared to all the treatments. The microstructure images of surimi confirmed that addition of CCP modified the ice crystal growth during frozen storage. This study suggests that CCP can be a potential additive to protect protein from denaturation along with partial replacement of chemical cryoprotectants.     

Validation of Xanthophyll Levels in Catfish Fillets Using Digital Photography

Catfish fillets with yellow coloration have become a prevalent problem for the catfish industry. This problem is due to the unacceptability in the market because the shift in fillet color is considered of lower quality by the consumers. To help the catfish industry better understand the yellow coloration of catfish fillet, a digital photography measurement method was developed to evaluate the yellowness. Sixty catfish fillets with a range of degrees of visible yellowness were taken directly from the processing line. The fillets were photographed in a light box with a digital camera. The photos were calibrated with the X-Rite ColorChecker standardized color target. CIELAB readings of the fillet photos were recorded, and the b* value was used to indicate the yellowness of catfish fillets. The xanthophyll levels of fillets were analyzed with high performance liquid chromatography. The actual xanthophyll level in catfish fillet was calculated as the sum of lutein, zeaxanthin, and alloxanthin. A linear correlation was found between the LAB b* values and xanthophyll levels of the 60 catfish fillets.     

日粮中鱼蛋白水解物对黄颡鱼生长、体成分和血清生理指标的影响

为了探讨鱼蛋白水解物对黄颡鱼生产性能的影响,以日本鳀粉为对照,以实用型黄颡鱼饲料配方模式为基础开展实验:1以30.5%鱼粉为对照(FM),在相同配方模式下,以6%鱼蛋白水解物(MPH6)替代20%的鱼粉;2以30.5%鱼粉为对照(FM),在无鱼粉日粮中分别添加3%(FPH3)、6%(FPH6)、12%(FPH12)鱼蛋白水解物;共设计5组等氮等能实验日粮,在池塘网箱中养殖黄颡鱼[初始体质量(30.08±0.35)g]60 d。结果显示:与FM相比,FPH12在SGR、FCR、PRR和FRR方面均无显著差异,而MPH6、FPH3、FPH6组SGR降低了15.45%~24.39%,FCR升高了32.14%~42.86%,MPH6、FPH6差异显著,在PRR和FRR方面,MPH6、FPH3、FPH6组PRR降低了21.11%~27.78%,MPH6组FRR降低了41.51%;全鱼水分、粗蛋白、粗脂肪和灰分各组间差异不显著,FPH3、FPH6、FPH12肌肉多种游离氨基酸水平显著高于FM,其中Thr、Val、His与其在日粮中的水平显著相关;FPH6组HSI显著低于FM,鱼蛋白水解物对CP、VSI、肠体比的影响不显著;血清AST、ALT、HDL、LDL、TP、CHOL、TG无显著差异,FPH3组ALB显著低于FM。研究表明:黄颡鱼日粮中,12%鱼蛋白水解物(干物质)与30.5%鱼粉在生长速度、饲料效率、血清生理指标等方面具有一定的等效性;过高的植物蛋白日粮影响了黄颡鱼的生产性能;饲喂鱼蛋白水解物日粮使黄颡鱼肌肉游离氨基酸的含量升高,特别是呈味氨基酸的含量增加。     

Design and development of a geothermal temperature control system for broodstock management of channel catfish Ictalurus punctatus

A control system was designed to raise and maintain water temperatures within 0.03-ha earthen ponds to a range conducive for spawning (24–30 °C) channel catfish Ictalurus punctatus. Heating was done during February 2001 to April 2001, when temperatures would have otherwise prohibited spawning (<24 °C). Temperature was increased from 10 °C (ambient) by 2 °C per day, and maintained at 27 °C, by the addition of geothermally warmed water (36 °C). The control system substantially increased the controllability and precision of heating ponds compared to manual operation. Systems were designed to control sets of four ponds. In designing this control system, consideration of biological constraints was essential. Reproduction in channel catfish is most strongly influenced by temperature. Because cold fronts are common during the winter and early spring (January–March), it was essential to ensure that pond temperatures did not fall below the range for spawning. Constraints on the heating rate and temperature variability to maintain fish health and stimulate spawning behavior were considered. Components of the control system included temperature measurement devices (type-T thermocouples), a central electronic control unit, electronic switches and electrically actuated ball valves. In response to the temperature sensed by each thermocouple, the controller sent a message to close or open the valve. When the valve was opened, warm water was added to the pond to increase the average pond temperature. Hardware and algorithm design and initial system testing were the major components of this project. The final design incorporated information on relevant biological parameters and safety features including peak pond temperature, independent aeration and water pressure control mechanisms. Initial results indicate successful control of this biological system, and ongoing studies suggest similar mechanisms may be used for additional control objectives. In particular, this system could be used to vary pond temperatures to study biological responses and to cool ponds by addition of well water during summer months.     

鲶鱼部分组织抗菌肽抗菌功效的研究

采用乙酸提取法提取鲶鱼表面黏液、皮肤、鱼肉、内脏等不同组织抗菌肽,采用双缩脲法测定蛋白质含量;以大肠杆菌、金黄色葡萄球菌和枯草芽孢杆菌为受试菌,采用滤纸片法,比较不同组织抗菌肽对不同菌种的抗菌效果,结果表明：各组织蛋白质含量分别为黏液0.336mg/mL、皮肤1.629mg/mL、鱼肉10.284mg/mL、内脏0.562mg/mL。内脏提取物的抑菌效果最好,且对枯草芽孢杆菌抑制效果最佳,抑菌圈直径达10.73mm。     

斑点叉尾(鱼回)LEAP2成熟肽在毕赤酵母中的表达及其抑菌活性

LEAP-2 (Liver expressed antimicrobial peptide-2) 抗菌肽是一类主要在动物肝脏中表达的小分子肽,其生物学活性来自成熟肽区域,它们在动物的免疫系统中发挥了重要作用。本文以斑点叉尾鮰 (Ictalurus punctatus) LEAP-2成熟肽(mLEAP2)为研究对象,拟通过建立毕赤酵母表达系统,实现mLEAP2在毕赤酵母中的重组DNA表达,以期获得具生物学活性的重组体mLEAP2 (rmLEAP2),为进一步研究其功能和扩大制备奠定基础。首先以既有的斑点叉尾鮰mLEAP2的原核表达载体“pET-32a-mLEAP2”为模板,通过PCR扩增,获得5’端含EcoRⅠ酶切位点、3’端含NotⅠ酶切位点及6×his标签的LEAP-2成熟肽基因mLEAP2,然后将其与真核表达载体pPIC9K连接,构建重组表达载体“pPIC9K-mLEAP2”;电转化至毕赤酵母GS115细胞中,经不同浓度G418和选择性培养基筛选,获得高拷贝酵母转化子;在29 ℃、250 r/m条件下,以0.5%的甲醇对其诱导表达120 h;采用固化金属离子亲和层析(IMAC)对表达产物进行纯化,通过MALDI-TOF/TOF质谱对纯化产物的结构进行分析和鉴定。结果表明：表达的重组体蛋白为预期的目的蛋白rmLEAP2,其表达量约2.2 mg/L;另一方面,抑菌实验结果显示rmLEAP2具有明显的抑制枯草芽孢杆菌(Bacillus subtilis)的活性。     

Emergence of Edwardsiella piscicida in Farmed Channel ♀, Ictalurus punctatus × Blue ♂, Ictalurus furcatus,Hybrid Catfish Cultured in Mississippi

There is a trend toward the increased incidence and prevalence of Edwardsiella piscicida septicemia in US catfish aquaculture, particularly in channel ♀, Ictalurus punctatus, × blue ♂, I. furcatus, hybrid catfish. From 2013 to 2017, a total of 3242 disease case submissions were made to the Aquatic Research and Diagnostic Laboratory (ARDL) at the Thad Cochran National Warmwater Aquaculture Center in Stoneville, MS. Of these, 1400 (43.2%) were hybrids. E. piscicida was suspected in 138 (4.3%) of cases, the majority of which (89.1%) were from hybrid catfish. A molecular survey of these isolates confirmed the majority (92.0%) to be E. piscicida. Furthermore, cases of E. piscicida from hybrids submitted to the ARDL and the Aquatic Diagnostic Laboratory of the Mississippi State University College of Veterinary Medicine in Starkville, MS, were documented for gross lesions and histological analysis. Grossly, E. piscicida presents with small dermal ulcerations, a raised fluid‐filled cranial midline lesion that is frequently ulcerated, hemorrhage in the gills, exophthalmia, and abdominal distension. Internally, lesions include splenomegaly, straw‐colored ascites, renomegaly, and occasionally hemorrhagic intestines. Histopathological examination is in agreement with gross observations, and infected fish repeatedly demonstrate a mononuclear meningoencephalitis, hemorrhagic branchitis, splenitis, ulcerative dermatitis, granulomatous interstitial nephritis, and hepatitis coupled with a hemorrhagic enteritis.     

Spatial and temporal variation in otolith chemistry and its relationship with water chemistry: Stock discrimination of Sperata aor

Long‐whiskered catfish, Sperata aor (Hamilton 1822), is commercially important in food, ornamental and sport fisheries. The fish is mainly caught from the wild populations because its aquaculture practices are not commercialised. Inland fishery in the Ganga basin is mostly unorganised; hence, no published report is available on the trend of S. aor production from the selected habitat. In India, S. aor has been categorised vulnerable mainly due to natural and anthropogenic threats. Otolith chemistry shows variation with changing physico‐chemical conditions of the fish habitat. Therefore, the present study was conducted with the objective to analyse spatio‐temporal variations in water chemistry in relation to environmental factors; relationship between water and otolith chemistry; and spatio‐temporal variations in otolith chemistry to discriminate the stocks of S. aor inhabiting the River Ganga. Most of the element: Ca ratios in water samples did not show significant correlations with environmental factors, viz. temperature and conductivity. Only few element: Ca concentrations in otoliths were positively correlated to their corresponding ratios in the ambient water. In the selected study area, the S. aor populations were discriminated into four stocks possibly because of heterogeneous water chemistry at the sampling sites, and physical barriers. In the present study, otolith chemistry showed relatively low temporal variability as compared to spatial variability; thus, the classification accuracy of individuals to their original populations remained consistent over the selected time period. The findings could be useful in devising scientifically sound management strategies and/or any conservation plans for the vulnerable S. aor populations inhabiting the River Ganga.