Development and evaluation of the polymerase chain reaction (PCR) method for diagnosis of spring viremia of carp virus (SVCV)

Aquaculture has been expanded rapidly to become a major commercial and food-producing sector worldwide in recent decade. In parallel, viral diseases rapidly spread among farms causing enormous economic losses. The accurate detection of pathogens at early stages of infection is a key point for disease control in aquaculture. Spring Viraemia of Carp Virus (SVCV) is a very severe pathogen of carp fishes in different parts of the world and is categorized as a reportable listed disease in the annual published list of World Organization for animal Health (OIE). The objective of this study was to develop and evaluate RT- PCR test for detecting SVC virus and also the sensitivity and specificity of this test. A semi nested RT- PCR was designed using combination of three primers: two external (SVCF , SVCR) and one internal (SVCS) primers which based on conserved region of G gen. The specificity of designed primers (only external ones) by examination on Viral Hemorrhagic Septicemia Virus (VHSV) and Infectious Hematopoietic Necrosis Virus (IHNV) was confirmed. For optimizing of the PCR test, primer concentration, primer annealing temperature, cycle number and Mgcl2 concentration were surveyed. Also for validity test, prevention of false negative and Assurance of its accuracy, a competitive internal control (mimic) designed and its suitable concentration was defined. Evaluation of the sensitivity of designed test were conducted first by comparing the different commercially available RNA isolation guidelines, two guidelines: isotiocyanate phenol–chloroform based protocols (RNX–Plus Iran, Iq2000 kit Taiwan ) and two column based protocols (Cinna pure RNA Iran , high pure viral RNA kit, Roche Germany ). The results indicated that the column based protocols (Roche method and Cinna pure), yield 36.77 ng/μl and 16/47 ng/μl RNA concentration respectively, which were significantly higher than other protocols(P<0.05). Then for evaluation of extracted RNA sensitivity, Serial dilution of SVCV strain 56.70 grown in EPC (1.9×105 TCID50/ml) was examined To compare sensitivity. Extracted RNA from serial dilution with stone's primers and commercial IQ-2000 kit were examined simultaneously. The result indicated that designed semi- nested RT- PCR was able to recognize SVC virus to 10-4 dilution and stone's primer recognize to 10-3 dilution whereas Iq-2000 commercial kit did not recognized in any dilution. In high virus titer in designed test two DNA band (462 bp and 266 bp) produced, and by decreasing virus titer 462 bp was omitted. In low virus titer or lack of virus, just DNA band (mimic) 729 bp can propagate. After designing and optimizing PCR test, a total of 400 suspected cultured Cyprinus carpio with high mortality from 4 aquaculture zone of Khuzestan province were collected and tested for SVCV during 2012- 2013 using developed PCR method and IQ- 2000. The results indicated that SVC virus was not observed in samples using both methods.

The statistical properties of recreational catch rate data for some fish stocks off the northeast U.S. coast

Recreational fisheries in the waters off the northeast U.S. target a variety of pelagic and demersal fish species, and catch and effort data sampled from recreational fisheries are a critical component of the information used in resource evaluation and management. Standardized indices of stock abundance developed from recreational fishery catch rates are routinely used in stock assessments. The statistical properties of both simulated and empirical recreational fishery catch-rate data such as those collected by the National Marine Fisheries Service (NMFS) Marine Recreational Fishery Statistics Survey (MRFSS) are examined, and the potential effects of different assumptions about the error structure of the catch-rate frequency distributions in computing indices of stock abundance are evaluated. Recreational fishery catch distributions sampled by the MRFSS are highly contagious and overdispersed in relation to the normal distribution and are generally best characterized by the Poisson or negative binomial distributions. The modeling of both the simulated and empirical MRFSS catch rates indicates that one may draw erroneous conclusions about stock trends by assuming the wrong error distribution in procedures used to developed standardized indices of stock abundance. The results demonstrate the importance of considering not only the overall model fit and significance of classification effects, but also the possible effects of model misspecification, when determining the most appropriate model construction.

Effect of chloral hydrate on metabolic rate of Labeo rohita (Ham) and Poecilia reticulata (Peters)

Comparative impact of chloral hydrate anaesthesia on the metabolic rate of Indian major carp Labeo rohita and larvivorous fish Poecilia reticulata was assessed. Observation on the Oxygen Consumption Rate (OCR) revealed that in common guppies OCR was substantially low (1.105 and 1.097 mg/g/hr) at 0.1 and 0.25 g/l concentrations of chloral hydrate as against OCR of 1.487 mg/g/hr in the control. Fry of L. rohita in group showed lower metabolic rates in the control as well as treated conditions as compared to the individuals of this fish. This may be due to sympathetic psychophysiological reflex of grouped fish. Higher dose of chloral hydrate (0.25 g/l) also caused higher OCR probably due to distress. Application of chloral hydrate also favoured lesser release of metabolic wastes (ammonia and carbon dioxide). There was significant positive correlation between time and oxygen consumption, whereas, for time and OCR this relationship was negative. Regression of chloral hydrate doses for OCR and time has also been calculated.