Antimicrobial susceptibility and enrofloxacin resistance of streptococcal bacteria from farmed Nile tilapia,Oreochromis niloticus (Linnaeus 1758) in Thailand

This study examined the antimicrobial susceptibility and mutation(s) in quinolone resistance‐determining regions (QRDRs) in streptococcal pathogens isolated from farmed Nile tilapia Oreochromis niloticus in Thailand. Surveillance of antimicrobial susceptibility in tilapia streptococcal pathogens reveals that Streptococcus agalactiae (n = 97) and Streptococcus iniae (n = 3) from diseased tilapia were susceptible to amoxicillin, florfenicol, sulfamethoxazole/trimethoprim and sulfadimethoxine/ormetoprim, however, only 78 isolates were susceptible to enrofloxacin. Twenty‐two enrofloxacin‐resistant S. agalactiae isolates were further examined for mutations in the QRDRs of gyrA, gyrB, parC and parE genes. Twenty isolates had single base pair changed in the gyrA sequence, C‐242‐T. Point mutations in gyrB, GC‐1135, 1136‐AA and T‐1466‐G, were identified in one isolate. All resistant isolates harboured a mutation in the parC gene, C‐236‐A, while no mutations were observed in the parE gene. The study represented mutations of gyrA and parC genes as marked modification of the enrofloxacin‐resistant S. agalactiae from farmed tilapia. This study is a primary report of the QRDRs mutations associated with fluoroquinolone resistance from streptococcal pathogen in the cultured fish. The phenotypic and genotypic characterization of enrofloxacin resistance S. agalactiae evident in this study has led to an improved regulation of antimicrobial use in Thai aquaculture.     

罗非鱼"突眼"病细菌性病原的分离与鉴定

从患"突眼"症状的罗非鱼体内分离到多株细菌,经人工感染证实其中TL60829NA菌株为病原菌,经生理、生化和API细菌鉴定系统以及分子生物学试验,鉴定其为痢疾志贺氏菌(Shigella dysenteriae)。其主要生理生化特征为革兰氏阴性短杆菌,不具动力,氧化酶、过氧化氢酶、V-P反应、吲哚试验、苯丙氨酸、甘露醇、KCN均为阴性;对氨苄西林、羧苄西林、红霉素、多西环素、氟哌酸、卡那霉素敏感,对呋喃唑酮、复方新诺明、磺胺和庆大霉素不敏感。     

罗非鱼鱼皮提取明胶的酶法脱脂工艺研究

用脂肪酶对罗非鱼鱼皮进行酶法脱脂工艺研究。通过单因素与正交试验确定最适的酶法脱脂工艺为:pH值9,脱脂时间3h,酶添加量2%,料液比1:4,温度40℃,该工艺的脱脂率可达67.5%。通过超声波辅助脱脂研究表明其脱脂率并无明显的提高。     

低温对罗非鱼基因组DNA甲基化的影响

以经过连续多代抗寒选育获得的尼罗罗非鱼耐寒品系为实验材料,采用DNA甲基化敏感扩增多态性(methylation sensitive amplified polymorphism,MSAP)分析方法从全基因组水平上探讨了低温适应对罗非鱼DNA序列中CCGG位点的甲基化水平的影响。结果显示,选用的18对选择性扩增引物共检测到849个位点。耐寒品系检测到的位点数为411个,对照组为438个,其中发生甲基化的位点数分别为72和104个,总甲基化率分别为17.52%和23.74%;全甲基化位点分别为37个和65个,全甲基化率分别为9.00%和14.84%;半甲基化位点分别为35个和39个,半甲基化率分别为8.52%和8.90%。结果分析表明,尼罗罗非鱼耐寒品系的总甲基化水平、全甲基化水平和半甲基化水平较对照组均有一定程度的下降。与对照组相比,尼罗罗非鱼耐寒品系基因组DNA甲基化总体水平下降了6.22%,其中以全甲基化位点变异为主,其下降幅度比例明显,为5.84%。由此可见,经过连续多代的低温胁迫可导致尼罗罗非鱼DNA甲基化水平发生改变,发生了去甲基化反应,表现为基因组甲基化程度降低的特征,说明了DNA甲基化与罗非鱼抗寒性反应密切相关。     

Generation of mouse monoclonal antibodies specific to tilapia immunoglobulin using fish immunoglobulin/BSA complex for monitoring of the immune response in Nile tilapia Oreochromis niloticus

The simple immunoprecipitation method was used to isolate tilapia immunoglobulin (Ig) for immunization in order to produce monoclonal antibodies (MAbs) specific to tilapia Ig. First, the tilapia antiserum against bovine serum albumin (BSA) was prepared by peritoneal injection of BSA into tilapia, and the tilapia anti‐BSA antiserum was used to precipitate BSA to form the Ig/BSA immune complex. The Ig/BSA immune complex was then injected into Swiss mice for hybridoma production. After fusion, three hybridoma clones producing MAbs specific to the tilapia antibody were selected by dot blot and Western blot. All MAbs (101A, 59G, and 11A) were bound specifically to the heavy chain of immunoglobulin M (IgM). The MAbs 101A and 59G demonstrated twofold higher affinity than MAb 11A and the commercialized antibody. However, MAbs 11A could also bind to the heavy chain of IgM in Asian seabass, Lates calcarifer, as well. These MAbs can be used to monitor the immune responses of individual fish by indirect ELISA upon exposure to various antigens.     

Relationship Between Dietary Protein Source and Feeding Frequency During Feeding Nile Tilapia,Oreochromis niloticus (L.) Cultured in Concrete Tanks

ABSTRACT

A 24-week feeding trial was conducted in concrete tanks with Nile tilapia, Oreochromis niloticus (L.) with an average initial weight and length of 50.87 ± 6.03 g and 14.4 ± 0.45 cm, respectively, to examine the effect of two protein sources and two feeding frequencies on growth performance, production traits, and body composition. Twelve 4 m³ concrete tanks (2 × 2 × 1.25 m, long, width, and height) were each stocked with 100 fish and fed diets containing either fish meal protein or soybean meal protein at two feeding frequency of two times daily or four times daily (2 × 2 × 3 factorial experiment). The results revealed that there was no significant effect of dietary protein sources on growth rate, whereas there was a significant increase in growth rate with increasing feeding frequency (P < 0.05). The same trend was also observed for mean body weight (g), production rate (kg/m3), specific growth rate (SGR % day^?1). The best final mean body weight (g), specific growth rate (SGR % day^?1), and production rate (kg/m3) were recorded in groups of fish fed with feeding frequency four times daily. Whole fish moisture, protein, fat, and ash contents were significantly influenced (P ≤ 0.05) by protein sources and feeding frequency, except protein not influenced with feeding frequency. Energy was significantly (P < 0.05) influenced by feeding frequency, but not by dietary protein sources. Economic evaluation indicated that soybean meal (Diet B) at four times feeding daily was the most cost-effective and affordable feed strategy for farmers. We conclude that a soybean diet fed four times daily is recommended for adult Nile tilapia reared in concrete tanks.     

超临界CO2干燥罗非鱼片的传质模型和数值模拟

为了探讨超临界CO2干燥罗非鱼的机理,该文基于微分床的质量守恒定律建立了超临界CO2干燥罗非鱼片的传质数学模型,通过Matlab软件对其干燥过程进行了数值模拟,分析了干燥室内超临界CO2中的溶质质量分数和罗非鱼片中的溶质质量分数随干燥时间和干燥床高度变化的规律,揭示溶质在超临界CO2干燥过程中的传质模式。结果表明:在超临界CO2干燥罗非鱼片的过程中,溶质传质主要以对流扩散为主,而以轴向扩散为辅;数值模拟的结果与试验值拟合良好(R2=0.97),相对偏差的绝对值在10%以内,说明建立的传质数学模型能较好的模拟超临界CO2干燥罗非鱼片的过程。研究结果可为超临界CO2干制罗非鱼片的工业化生产的过程控制提供参考。     

不同生长阶段尼罗罗非鱼IGF1基因表达分析

为阐明IGF1基因在罗非鱼生长过程中的表达规律,为罗非鱼的分子选育等工作奠定了理论基础,本实验利用实时荧光定量PCR技术跟踪尼罗罗非鱼(Oreochromis niloticus)在不同生长阶段（0、20、40、60、80、125、145、165、185、205天）肝脏和肌肉胰岛素样生长因子1(IGF1)基因表达的发育性变化。结果表明：实验鱼的体重增长过程可以分为快速生长期和平稳期。在快速生长期时,肝脏IGF1基因表达水平较高,平稳期时较低,肝脏中IGF1基因表达情况与体重增长趋势基本吻合。肌肉中IGF1基因mRNA含量要低得多,而且表达模式与肝脏IGF1基因不相同。研究结果提示：IGF1 mRNA的表达具有明显的时空变化和组织特异性。肝脏中IGF1基因的表达量变化与罗非鱼体重增长趋势一致。肌肉中IGF1表达模式与肝脏中不同,提示肌肉中IGF1的分泌和作用有其自身的特异性。     

解冻损失约束下罗非鱼冻结效率控制参数优化研究

速冻方式不同对鱼肉温降速率、均匀性以及解冻后品质的影响不同,最佳速冻方式选取已成为行业内提升鱼类速冻效率与维持鱼肉品质的主要方式之一。以罗非鱼为例,结合3D扫描逆向建模与计算流体力学数值模拟等技术直观获取冷风式速冻、氯化钠和氯化钙浸渍速冻方式下罗非鱼温度时空分布规律,并通过与实验值比较验证模拟结果的准确性与可靠性,综合对比分析不同速冻方式对罗非鱼冻结速率、均匀性以及水分流失的影响规律,确定最佳速冻方式并提出优化设计方案。研究结果表明,温度模拟值与实测值最大偏差为1.81℃,最大均方根误差与平均绝对百分比误差分别为1.017℃、18.9%,且冻结过程罗非鱼温度降至-15℃时,3种速冻方式所需冻结时间模拟值与实测值最大相对偏差仅为6.89%。其次,氯化钠和氯化钙两种浸渍速冻方式下罗非鱼冻结速率基本相同,相比冷风式速冻冻结时间约降低73%,冻结均匀性提升10~40倍。两种浸渍速冻方式下罗非鱼解冻后水分流失率仅为3.89%和3.92%,分别比冷风式速冻方式降低10.37%和9.68%左右。最后,提出了一种氯化钠液流化速冻方式优化方案,通过综合考虑冻结速率、均匀性及能耗确定最佳流速为2.5m/s,且相比鱼体头朝入口水平放置与竖直放置,鱼体背朝入口水平摆放方式的冻结效率最优。