Genome analysis of Betanodavirus from cultured marine fish species in Malaysia

Betanodavirus is the causative agent of the viral nervous necrosis (VNN) or viral encephalopathy and retinopathy disease in marine fish. This disease is responsible for most of the mass mortalities that occurred in marine fish hatcheries in Malaysia. The genome of this virus consists of two positive-sense RNA molecules which are the RNA1 and RNA2. The RNA1 molecule contains the RdRp gene which encodes for the RNA-dependent RNA polymerase and the RNA2 molecule contains the Cp gene which encodes for the viral coat protein. In this study, total RNAs were extracted from 32 fish specimens representing the four most cultured marine fish species in Malaysia. The fish specimens were collected from different hatcheries and aquaculture farms in Malaysia. The RNA1 was successfully amplified using three pairs of overlapping PCR primers whereas the RNA2 was amplified using a pair of primers. The nucleotide analysis of RdRp gene revealed that the Betanodavirus in Malaysia were 94.5-99.7% similar to the RGNNV genotype, 79.8-82.1% similar to SJNNV genotype, 81.5-82.4% similar to BFNNV genotype and 79.8-80.7% similar to TPNNV genotype. However, they showed lower similarities to FHV (9.4-14.2%) and BBV (7.2-15.7%), respectively. Similarly, the Cp gene revealed that the viruses showed high nucleotide similarity to RGNNV (95.9-99.8%), SJNNV (72.2-77.4%), BFNNV (80.9-83.5%), TPNNV (77.2-78.1%) and TNV (75.1-76.5%). However, as in the RdRp gene, the coat protein gene was highly dissimilar to FHV (3.0%) and BBV (2.6-4.1%), respectively. Based on the genome analysis, the Betanodavirus infecting cultured marine fish species in Malaysia belong to the RGNNV genotype. However, the phylogenetic analysis of the genes revealed that the viruses can be further divided into nine sub-groups. This has been expected since various marine fish species of different origins are cultured in Malaysia.     

Histochemical and morphometric studies of the cranial tibial muscle in dogs of different dynamic aptitudes (greyhound, German shepherd and fox terrier)

A histochemical and morphometric study of the cranial tibial muscle of the greyhound, the German shephard and the fox terrier was carried out using hematoxyline-eosin, PAS, Gomori trichrome, SDH, LDH, NADH-tr and ATPase staining (with pH adjustments to both acidic and basic levels), with the goal of comparing the proportion and distribution of type I and type II fibers within the muscle. Significant differences were found between the three breeds of dogs. It is suggested that these differences are related to the locomotor activities of the three breeds.     

全球鱼类和水产品前瞻

1市场和价格鱼类及鱼产品价格总体下行。由于需求减少．只有那些产量供应紧张的品种才幸免于这场价格下跌风暴．包括野生白鱼和黄鳍金枪鱼、养殖三文鱼和罗非鱼．价格仍趋稳定。其他养殖品种．如欧洲鲷和大菱鲆价格创历史新低,原因是产量大幅增长。另一方面．越南鲶鱼价格尽管在2008年初下跌．不过正在恢复中上涨。     

Histochemical and morphometric characterization of broncho-pneumonia in calves caused by infection with Mycoplasma bovis

The aim of this study was to identify morphometric histological features of pneumonia caused by Mycoplasma bovis in calves. Eight three-month-old calves were infected with M. bovis and samples of their lung tissue, three weeks after exposure, compared to samples from four uninfected calves. In the uninfected animals the goblet cells were clustered in the crypt area of the epithelial folds, while in the infected calves they had migrated towards the tips of the folds and were distributed evenly throughout the folds. In infected lung tissue there was goblet cell hyperplasia and metaplasia in the bronchioles and an increased epithelial height. Goblet cell mucin in uninfected calves was acidic, but in infected calves most goblet cells contained neutral mucins. These morphometric and histochemical bronco-epithelial changes may be able to be used as markers of the severity of bovine respiratory mycoplasmosis.     

Animal welfare aspects of the killing of fish

According to the Animal Protection Law (1986) fish are to be killed by methods which do not cause pain. However, the regulations do not cover the killing of non-food fish. A questionnaire, conducted among 85 fish scientists, revealed that single fish should be killed by a blow on the head, and larger numbers by electrical methods or by use of chemicals. Decapitation was proposed for eels. A regulation from 1936 stipulates the methods for the slaughtering of food fish. Mechanical or electrical stunning is compulsory except for eel and flatfish. The questionnaire showed that in general the present legal regulations are sufficient for the slaughtering of fish with the exception of eels. The commercially available apparatus for stunning and killing do not always fulfill the requirements of animal protection, slaughtering technology and safety for the user. Official testing of these apparatus as well as the evaluation of new methods--like CO2-stunning--are necessary in order to prevent the use of methods which are feasible, but do not fulfill animal welfare, especially for eel.     

鱼油在水产饲料中的应用

鱼油是湿法加工鱼粉过程中的副产品,是将海洋鱼类及其废弃物经蒸馏、压榨和精炼而得到,以脂肪形式存在于鱼肉中或以分散油形式存在于鱼体内的油脂。鱼油的主要成分是甘油三脂,此外还含有少量类脂、磷甘油醚、脂溶性维生素以及蛋白质降解物等,其中磷脂主要由磷脂酰胆碱和磷脂酰乙醇胺组成。与一般动植物油脂相比,鱼油中大部分脂肪酸具有较长的碳链和多元不饱和度,长链脂肪酸通常占总脂肪量的1/4￣1/3,有时高达1/2,其中5￣6个双键的脂肪酸通常占鱼油的15%￣30%,尤其以n-3系列的高度不饱和脂肪酸(H U FA),如二十碳五烯酸(E PA)和二十二碳六…     

Effect of heat-inactivated fish and non-fish derived probiotics on the innate immune parameters of a teleost fish (Sparus aurata L.)

The in vitro effects of four heat-inactivated bacterial species on the cellular innate immune responses of the gilthead seabream (Sparus aurata L.) were investigated. Head-kidney leucocytes were isolated and incubated for 30 min with two bacteria isolated from seabream skin (Pdp11 and 51M6; members of the Vibrionaceae and in the genus Shewanella) and two bacteria used as probiotics in humans and cattle (Lactobacillus delbrüeckii subsp. lactis and Bacillus subtilis) at 5 x 10(5), 5 x 10(6) or 5 x 10(7)CFU/ml. After incubation, different cellular innate immune parameters (leucocyte peroxidase content, phagocytosis, respiratory burst activity and cytotoxicity) were evaluated. The leucocyte peroxidase content was significantly higher after incubation with 51M6 at 5 x 10(7)CFU/ml. Head-kidney phagocytes were able to engulf the four bacterial species in all four cases, L. delbrüeckii subsp. lactis and B. subtilis being the most actively phagocytized. The incubation of seabream leucocytes with 51M6, L. delbrüeckii subsp. lactis and B. subtilis resulted in a great increase in respiratory burst activity. Cytotoxic activity was generally stimulated in a dose-dependent manner, the enhancement obtained with 5 x 10(7)CFU/ml being statistically significant. The usefulness of in vitro assays for screening and selecting candidate probiotic bacteria, as well as for optimising their effective dose, is discussed in relation with their immune-modulatory properties.     

Radiodiagnostic examination of the swimbladder of some fish species

Radiodiagnostic methods have not been used previously for studying the anatomy and diseases of the swimbladder of freshwater fish species. In this study, the radiographic anatomy of the swimbladder and species-related differences in swimbladder structure were studied on plain radiographs taken of 12 Hungarian fish species of major economic importance. Changes observed by radiography were also studied by conventional parasitological methods. The radiodiagnostic method reported here appears to be a useful complement to diagnostic examinations that have been based merely on dissection so far. It enables evaluation of the pathological lesions in live condition, without causing damage to the fish.     

Occurrence and health aspects of PCB congeners in freshwater fish

6 PCB-Congeners, being ruled by the regulatory limits from 23. 3. 1988, are considered to be representative under analytical aspects. The additional consideration of the two further congeners 77 and 118 is recommended under toxicological aspects. A widespread survey is given over the pollution status of marine fish, freshwater fish, and belonging to products. In the years between 1986 to 1988 2229 specimens were analysed for their contents of 8 PCB-Congeners and 6 important chlorinated hydrocarbons. Contents in marine fish were at least 10 times, mostly 100 times lower than regulatory limits. In this area contamination is more influenced by species than by catching grounds. Regulatory limits can however be exceeded in freshwater fish from severely polluted main rivers, where contents are more influenced by catching grounds than by species.     

Cloning and expression of Toll-like receptors 1 and 2 from a teleost fish, the orange-spotted grouper Epinephelus coioides

The two Toll-like receptors, TLR1 and TLR2 were cloned from orange-spotted grouper, Epinephelus coioides, an important teleost fish in mariculture of Asia. The cDNA sequences of TLR1 and TLR2 are 3195 and 3439 bp long, with an open reading frame (ORF) of 2406 and 2466 bp, encoding 801 and 821 amino acids, respectively. The TLR family motifs, i.e. leucine-rich repeat (LRR) domains and Toll/interleukin (IL)-1 receptor (TIR) domains are conserved in the TLR1 and TLR2, with nine and ten leucine-rich repeat (LRR) domains, and with one TIR domain, respectively. The TLR1 and TLR2 had a constitutive expression in examined organs/tissues of na?ve orange-spotted grouper, and an increased expression of TLR1 and TLR2 at mRNA level was observed in immune organs such as in spleen of LPS and Poly(I:C) stimulated fish. An increased expression of TLR1 and TLR2 was also recorded in immune organs of the fish injected with the bacterial pathogen, Vibrio alginolyticus. Similarly, a significant rise in the expression of MyD88, an adaptor molecule which forms signalling complex with intracellular TIR domain, thus leading to the production of pro-inflammatory cytokines, such as IL-1β, was also observed in the LPS- and Poly(I:C)-stimulated, and V. alginolyticus-infected fish, indicating the possible role of TLR1 and TLR2 in the MyD88 signalling pathway. However, the mechanism involved in the increased expression of TLR1 and TLR2 following LPS and Poly(I:C) stimulation is at present unknown in fish, and further research should be carried out to identify ligands of fish TLR1 and TLR2 in order to understand the function of these receptors.     

Comparison of chloroform-methanol-extracted and solvent-free triglyceride determinations in four fish species

Lipids, including triglycerides, are important variables in fish bioenergetics and can be used to estimate overall fish condition. Triglycerides are the major energy storage form in fish and therefore are a more ecologically and physiologically relevant measure of bioenergetics than total lipids. Chloroform-methanol-extracted total body lipids (Bligh and Dyer) and total body triglycerides determined in chloroform-methanol extracts and unextracted whole-body fractions were measured in four fish species: northern pike Esox lucius, burbot Lota lota, slimy sculpin Cottus cognatus, and spottail shiners Notropis hudsonius. Determinations of total body lipids were consistently greater than those of total body triglycerides when measured in the same solvent-extracted fraction, although both measures followed similar trends. In an effort to eliminate the need for extraction with organic solvents, we compared the performance of an enzyme-based triglyceride assay in both the solvent-extracted fraction and a whole-body unextracted homogenate for each fish. The chloroform-methanol-extracted triglyceride values were consistently lower than triglycerides measured in the unextracted whole-body homogenate. In addition, comparison of triglyceride measurements revealed limitations to the solvent extraction and subsequent triglyceride determinations in lean fish. Thus, in addition to being simple, rapid, and not requiring organic solvents, determination of triglycerides in an unextracted whole-fish homogenate may be a useful alternative to chloroform-methanol-based methods of lipid extraction and subsequent triglyceride measurement.     

用POR和POR-RFLP方法检测和鉴定进口海鱼中的异尖线虫幼虫

应用PeR和PCR—RFLP方法对厦门口岸进口的各种海鱼中的异尖线虫幼虫进行检测鉴定。结果在带鱼、竹荚鱼、金线鱼、大眼鲷和白姑鱼等海鱼中发现了简单异尖线虫、典型异尖线虫、对盲囊线虫和针回线虫等4种异尖线虫。带鱼和竹荚鱼的异尖线虫感染率达100％,竹荚鱼异尖线虫感染强度最高,白姑鱼感染率和感染强度相对较低。少数海鱼同时感染2种以上异尖线虫,从带鱼中同时检出简单异尖线虫和针蛔线虫,从竹荚鱼中检出简单异尖线虫、对盲囊线虫和典型异尖线虫。内切酶实验结果表明,应用限制性内切酶UinfI有效鉴别上述4种异尖线虫。     

不同草种单播草坪蒸散量的研究

对草地早熟禾、多年生黑麦草、高羊茅、狗牙根、结缕草和野牛草6种常见草坪草草坪的蒸散量进行了研究,发现在充分灌水的条件下,冷地型草坪的蒸散量显著大于暖地型草坪(P<0.01);3种冷地型草坪间差异较小;暖地型草坪中狗牙根和结缕草的蒸散量显著高于野牛草(P<0.01)。限制灌水条件下,6种草坪的蒸散量差异均显著(P<0.05或0.01),其顺序为:高羊茅>早熟禾>黑麦草>狗牙根>结缕草>野牛草。通过充分灌水和限制灌水2个处理确定了6种草坪的年需水量范围。在不同水分梯度下,高羊茅的蒸散量随土壤含水量的减少而减少;狗牙根的蒸散量随土壤含水量的减少呈现降低的趋势。高羊茅和狗牙根蒸散量在一天内随温度的升高而增加,但温度维持在较高时由于气孔关闭导致蒸腾降低从而蒸散量降低。     

不同甘草种质材料营养价值研究

对6个不同甘草种质材料的异黄酮、水分、灰分、粗蛋白、粗脂肪、粗纤维和无氮浸出物含量进行了测定。结果表明,6个甘草的营养成分不同,其中,胀果甘草的无氮浸出物50.29%,根中总异黄酮含量891.8μg/g,均最高,乌拉尔甘草茎叶异黄酮含量、粗蛋白与粗脂肪含量较高。利用灰色关联度分析法对6个甘草进行综合评价得出,乌拉尔甘草营养价值最优,其次排序为胀果甘草粗毛甘草云南甘草光果甘草黄甘草。