Study of infectious haematopoietic necrosis disease in some hatcheries and farmed rainbow trout in Iran using immunohistochemical technique and to comparing the obtained results with the PCR works

To detect rainbow trout hatcheries for infectious hematopoietic necrosis virus, samples of kidney, liver, spleen, muscle, intestine, heart and gills of trout larvae were obtained from a number of trout hatcheries from different provinces. Also tissue samples were obtained for molecular works using RT- PCR procedure. Tissue samples were processed using standard histotechnique and the obtained sections were stained using immunohistochemical procedure. From 100 examined samples 35 were positive for IHN by immunohistochemical test. Also, from 100 samples examined, 43 were positive in RT- PCR studies. The obtained results show that some rainbow trout hatcheries are contaminated in different regions of country. Therefore, a definition of prevention and eradication criteria are now critical to protect the unaffected areas within the country.

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.

Mersey & Weaver Fisheries Advisory Committee 19th January, 1977.

This is the report from the Mersey and Weaver Fisheries Advisory Committee meeting, which was held on the 19th January, 1977. The report contains information the construction of a new surface water drain to Pennington Flash, fisheries activities, fish stocking, eel fishing and estuarial waters, and infectious Pancreatic Necrosis. The section on fisheries activities includes brown/rainbow trout, roach, perch, bream, carp, tench and rudd stocking and fish mortalities. The section on Infectious Pancreatic Necrosis highlights its prevention on imports of salmonid fish and ova. The Fisheries Advisory Committee was part of the Regional Water Authorities, in this case the North West Water Authority. This preceded the Environment Agency which came into existence in 1996.

Lune, Wyre & Furness Fisheries Advisory Committee 17th January, 1977.

This is the report from the Lune, Wyre and Furness Fisheries Advisory Committee meeting, which was held on the 17th January, 1977. The report contains sections on angling information on the River Lune, fisheries activities, fish stocking, eel fishing in estuarial waters, and Infectious Pancreatic Necrosis which highlights its prevention on imports of salmonid fish and ova. The section on fisheries activities includes salmon and sea trout propagation in Middleton hatchery; fishing conditions/monitoring; and a proposed fish counter in the River Wyre. The Fisheries Advisory Committee was part of the Regional Water Authorities, in this case the North West Water Authority. This preceded the Environment Agency which came into existence in 1996.

South Lancashire Fisheries Advisory Committee 12th January 1977.

This is the report from the South Lancashire Fisheries Advisory Committee meeting, which was held on the 12th January 1977. The report contains sections on fish mortalities at Henthorn on the 10th July, 1976, fisheries activities, fish stocking, eel fishing on estuarial waters, and infectious Pancreatic Necrosis highlighting its prevention on imports of salmonid fish and ova. The section on fish mortalities looks at fish deaths on the River Ribble caused by a discharge from Clitheroe Sewage Works. The report on fisheries activities includes information on artificial propagation and numbers of fish passing through Waddow Weir, Winckley Hall and Locks Weir between January to November 1976. The Fisheries Advisory Committee was part of the Regional Water Authorities, in this case the North West Water Authority. This preceded the Environment Agency which came into existence in 1996.

Eden & District Fisheries Advisory Committee 11th January, 1977.

This is the report from the Eden and District Fisheries Advisory Committee meeting, which was held on the 11th January, 1977. The report contains sections on netting on the Solway Firth, fisheries activities, fish stocking, eel fishing on estuarial waters, and infectious Pancreatic Necrosis. The section on fisheries activities includes general information on the River Eden and Esk; fish transfers; predators; disease; fish mortalities; restocking; and information of Holmwrangle hatchery. The section on Infectious Pancreatic Necrosis looks at this fish disease highlighting its prevention on imports of salmonid fish and ova. The Fisheries Advisory Committee was part of the Regional Water Authorities, in this case the North West Water Authority. This preceded the Environment Agency which came into existence in 1996.

South & West Cumberland Fisheries Advisory Committee 10th January, 1977.

This is the report from the South and West Cumberland Fisheries Advisory Committee meeting, which was held on the 10th January, 1977. The report contains information on water resources development in West Cumbria, Branthwaite Weir regarding the construction of a fish pass, fisheries activities, fish stocking, eel fishing on estuarial waters, and infectious Pancreatic Necrosis which highlights its prevention on imports of salmonid fish and ova. The section on fisheries activities includes general information on the River Derwent; diseases; fish mortalities; Holmwrangle hatchery and land drainage works. The Fisheries Advisory Committee was part of the Regional Water Authorities, in this case the North West Water Authority. This preceded the Environment Agency which came into existence in 1996.

Pathogens after shrimp: A rogue's gallery of the industry's four most destructive adversaries

The paper discusses the four most destructive shrimp pathogens, such as MBV, the monodon baculovisrus, IHHNV, the infectious hypodermal and hematopoietic necrosis virus, Vibrio harveyi, the luminous bacteria, and WSBV, the white spot syndrome-associated baculovirus. The effects, detection method and treatment for the four pathogens were also briefly discussed.

White spot syndrome virus (WSSV) transmission risk through infected cooked shrimp products assessed by polymerase chain reaction (PCR) and bio-inoculation studies

The aim of the study was to evaluate the resistance of white spot syndrome virus (WSSV) in shrimps (Penaeus monodon) to the process of cooking. The cooking was carried out at 1000C six different durations 5, 10, 15, 20, 25 and 30 min. The presence of WSSV was tested by single step and nested polymerase chain reaction (PCR). In the single step PCR, the primers 1s5 & 1a16 and IK1 & IK2 were used. While in the nested PCR, primers IK1 &IK2 – IK3 & IK4 were used for the detection of WSSV. WSSV was detected in the single step PCR with the primers 1s5 and 1a16 and the nested PCR with the primers IK1 and IK2 – IK3 & IK4 from the cooked shrimp samples. The cooked shrimps, which gave positive results for WSSV by PCR, were further confirmed for the viability of WSSV by conducting the bio-inoculation studies. Mortality (100%) was observed within 123 h of intra-muscular post injection (P.I) into the live healthy WSSV-free shrimps (P. monodon). These results show that the WSSV survive the cooking process and even infected cooked shrimp products may pose a transmission risk for WSSV to the native shrimp farming systems.

An epizootic of infectious hepatopancreatic and lymphoid organ necrosis disease in cultured Penaeus monodon: a case study

Shrimp disease of viral origin have caused large production losses worldwide. This paper presents a case study of shrimp (Penaeus monodon; Penaeus indicus) epizootic disease, covering an area of 1,050 ha in Andhra Pradesh, India. The disease struck shrimp farms in the area in July 1994. Samples from 26 shrimp farms were studied in the laboratory, and the pattern of the disease and of mortality recorded. The disease was classified as infectious hepatopancreatic and lymphoid organ necrosis disease (IHLN).

Rapid detection of white spot syndrome virus (WSSV) of Penaeus monodon by latex agglutination test using monoclonal antibodies

Latex beads were sensitized with monoclonal antibodies (MAb) rose against VP28 of WSSV. The optimum concentration of MAb required to sensitize the latex beads was 125 µg/ml. The sensitized latex beads were used to detect WSSV from PCR-positive stomach tissue homogenates obtained from infected shrimp. Stomach tissue homogenates from WSSV-infected shrimp agglutinated the sensitized latex beads within 10 minutes, while uninfected samples did not produce any agglutination, although non-specific agglutinations were observed in some samples. The analytical sensitivity, analytical specificity, diagnostic sensitivity and diagnostic specificity of the (LAT) agglutination test were assessed. The analytical sensitivity of the test was 40 ng of purified WSSV (2 µg/ml). The sensitized latex beads did not agglutinate with normal shrimp tissue or MBV-infected tissue homogenate. The test has a diagnostic sensitivity of 70 and 45%, respectively, compared to single-step and nested PCR. The diagnostic specificity of the test was 82%. This test is a simple and rapid on-farm test which can be used to corroborate clinical signs for the detection of WSSV in grow-out ponds.