Detection of viral diseases in shrimp Litopenaeus vannamei in Iran with emphasis on white spot disease prevention by using marine alga extract

Nowadays, following was expanded shrimp breeding and culture; viral diseases have been main problem which threatened shrimp industry in the country. Therefore, shrimp samples were obtained from different stages of Litopenaeus vannmei life cycle (larval, post larval, juveniles, adults and broodstocks) based on clinical signs in the breeding center and shrimp farming from Bushehr, Khozestan and Sistan and Baluchestan provinces. Viral diseases were detected by PCR (Polymerase Chain Reaction), histopathology and transmission electron microscopy (TEM) methods. Results of the PCR were indicated present white spot virus (WSV) in juveniles, sub adults and adults shrimp with medium intensity from three provinces, but it was not showed in larval and post larval stages. Histopathological sections were indicated hypertrophy and basophilic Cowdry type A formation in nucleus cells of gill, haematopoietic, lymphoid and epithelial's cuticles and intestinal tissues which was associated with small vacuoles increased in B cells of hepatopancreas tissue of infection shrimps. Transmission electronic microscopic studies were demonstrated that the length and diameter virus was detected, respectively, 300 ± 20 nm and 75 ± 5 nm. Considerable, results of the PCR were only displayed IHHNV in juvenile, adult and broodstock shrimps from breeding and farming center of Bushehr province. The main lesion pathology was formed eosinophilic Cowdry type A in nucleus cells of gill, haematopoietic, lymphoid and epithelial's cuticles and intestinal tissues. Whereas penaeid shrimps are lack specific immune system, hence, in the present study was used of marine alga (Lurensia snideria) collected from along costal Persian Gulf of Bushehr province for viral diseases were prevented. Powder alga extract were added with a ratio of 1 % to shrimp diet. Total haemocyte count (THC) and total protein plasma (TPP) were increased after 5 days of oral administration diets. When shrimps were infected by with spot virus experimentally, THC and TPP gradually were increased in both two groups (shrimps fed with diet containing alga extract and without alga extract) after 48h. Nevertheless; THC, TPP and survival of shrimp fed with diet containing alga extract were more than shrimp control in 15 days. So, oral administration Lurensia snideria extract was capable prevention infected L. vannamei via stimulant specific immune system.

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.

Hydrocarbon sensors for oil spill prevention and response, Seward, Alaska, April 8th-10th, 2008: workshop proceedings

During April 8th-10th, 2008, the Aliance for Coastal Technology (ACT) partner institutions, University of Alaska Fairbanks (UAF), Alaska SeaLife Center (ASLC), and the Oil Spill Recovery Institute (OSRI) hosted a workshop entitled: "Hydrocarbon sensors for oil spill prevention and response" in Seward, Alaska. The main focus was to bring together 29 workshop participants-representing workshop managers, scientists, and technology developers - together to discuss current and future hydrocarbon in-situ, laboratory, and remote sensors as they apply to oil spill prevention and response. [PDF contains 28 pages] Hydrocarbons and their derivatives still remain one of the most important energy sources in the world. To effectively manage these energy sources, proper protocol must be implemented to ensure prevention and responses to oil spills, as there are significant economic and environmental costs when oil spills occur. Hydrocarbon sensors provide the means to detect and monitor oil spills before, during, and after they occur. Capitalizing on the properties of oil, developers have designed in-situ, laboratory, and remote sensors that absorb or reflect the electromagnetic energy at different spectral bands. Workshop participants identified current hydrocarbon sensors (in-situ, laboratory, and remote sensors) and their overall performance. To achieve the most comprehensive understanding of oil spills, multiple sensors will be needed to gather oil spill extent, location, movement, thickness, condition, and classification. No single hydrocarbon sensor has the capability to collect all this information. Participants, therefore, suggested the development of means to combine sensor equipment to effectively and rapidly establish a spill response. As the exploration of oil continues at polar latitudes, sensor equipment must be developed to withstand harsh arctic climates, be able to detect oil under ice, and reduce the need for ground teams because ice extent is far too large of an area to cover. Participants also recognized the need for ground teams because ice extent is far too large of an area to cover. Participants also recognized the need for the U.S. to adopt a multi-agency cooperation for oil spill response, as the majority of issues surounding oil spill response focuses not on the hydrocarbon sensors but on an effective contingency plan adopted by all agencies. It is recommended that the U.S. could model contingency planning based on other nations such as Germany and Norway. Workshop participants were asked to make recommendations at the conclusion of the workshop and are summarized below without prioritization: *Outreach materials must be delivered to funding sources and Congressional delegates regarding the importance of oil spill prevention and response and the development of proper sensors to achieve effective response. *Develop protocols for training resource managers as new sensors become available. *Develop or adopt standard instrument specifications and testing protocols to assist manufacturers in further developing new sensor technology. *As oil exploration continues at polar latitudes, more research and development should be allocated to develop a suite of instruments that are applicable to oil detection under ice.

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.

Harmful algal blooms in coastal waters: options for prevention, control and mitigation

This report is the product of a panel of experts in the science of blooms of unicellular marine algae which can cause mass mortalities in a variety of marine organisms and cause illness and even death in humans who consume contaminated seafood. These phenomena are collectively termed harmful algal blooms or HABs for short. As a counterpart to recent assessments of the priorities for scientific research to understand the causes and behavior of HABs, this assessment addressed the management options for reducing their incidence and extent (prevention), actions that can quell or contain blooms (control), and steps to reduce the losses of resources or economic values and minimize human health risks (mitigation). This assessment is limited to an appraisal of scientific understanding, but also reflects consideration of information and perspectives provided by regional experts, agency managers and user constituencies during three regional meetings. The panel convened these meetings during the latter half of 1996 to solicit information and opinions from scientific experts, agency managers and user constituencies in Texas, Washington, and Florida. The panel's assessment limited its attention to those HABs that result in neurotoxic shellfish poisoning, paralytic shellfish poisoning, brown tides, amnesic shellfish poisoning, and aquaculture fish kills. This covers most, but certainly not all, HAB problems in the U.S.

Ciguatera Fish Poisoning: treatment, prevention and management

Ciguatera Fish Poisoning (CFP) is the most frequently reported seafood-toxin illness in the world, and it causes substantial physical and functional impact. It produces a myriad of gastrointestinal, neurologic and/or cardiovascular symptoms which last days to weeks, or even months. Although there are reports of symptom amelioration with some interventions (e.g. IV mannitol), the appropriate treatment for CFP remains unclear to many physicians. We review the literature on the treatments for CFP, including randomized controlled studies and anecdotal reports. The article is intended to clarify treatment options, and provide information about management and prevention of CFP, for emergency room physicians, poison control information providers, other health care providers, and patients.

Phosphate treatment of frozen prawns. Pt. 1. Screening of various phosphates for prevention of drip loss

Various phosphates and their mixtures were screened for their efficiency of preventing drip loss in frozen prawns. The effectiveness of the phosphates decreased in the following order: Sodium tripolyphosphate — Sodium pyrophosphate — Sodium hexametaphosphate Sodium metaphosphate — Sodium dihydrogen phosphate; the last two being ineffective. Even though thaw drip loss was reduced by the above treatments the organoleptic quality of the thawed as well as cooked products was unsatisfactory, discoloration being the major defect. A solution of a mixture of 12% sodium tripolyphosphate and 8.6% sodium dihydrogen phosphate or 2% citric acid in water when used as dip prevented thaw drip loss, improved cooked yield and organoleptic quality without adversely affecting the biochemical characteristics. Commercial scale trials showed that the results are highly reproducible.

Blackening of canned prawn and its prevention

The paper gives an account of the types of blackening associated with canned prawn in brine and their control. It was found that blackening caused by iron sulphide could be controlled by maintaining proper titratable acidity of fill brine in cans. The paper also elaborates on the factors responsible for or governing this critical titratable acidity. In regard to copper sulphide blackening, control was found to be difficult by maintaining the acidity or by additives such as EDTA when the copper content in the material went above the critical level.

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.

Removal of fungus and prevention of fungal reinfestation in stored dry salted fish

A simple and effective method is described for converting dry fish products infested with fungus and red halophilic bacteria to acceptable ones. Washing, drying and then smearing with a mixture of 3% sodium propionate in dry, refined salt in a ratio of 1 part mixture: 10 parts dry fish resulted in the fish having a shelf life of 5 months.

Prevention of infestation by Dermestes maculatus Degeer in east African dried fish by solar treatment

East African sun-dried fish infested by Dermestes maculatus were exposed to tropical sunlight at ambient temperature and analysed for insect mortality and weight losses. Solar treatment for 6 to 8 lo was highly effective for one layer of split sun-dried fish and 100% insect mortality was toted, while pest species were still present in the four layer batch. Weight losses between 1.2% and 10.2% were recorded, the top layer suffering the highest loss. The high surface temperature of 60°C caused fish to become brittle and quality losses occurred. A reduction in length of exposure/temperature is probably a presupposition for application of the method to local conditions.

Prevention of infestation by Dermestes maculatus Degeer in east African dried fish using pyrethrum and piperonyl butoxide

East African sun-dried fish dipped for 4 seconds in different solutions of pyrethrum and piperonyl butoxide were analysed for insecticide residue limits. All analyses showed residues above the FAO/WHO MRL; exceeding factors of between 7.6 (22.9 ppm) and 1.6 (5.3 ppm) were found for pyrethrum while exceeding factors between 5.1 (102 ppm) and 1.7 (33.1 ppm) were common for piperonyl butoxide after 6 months storage at ambient temperature. All insecticide treated fish, regardless of dip concentration, were observed to be less susceptible to infestation by Dermestes maculatus than samples of untreated fish. No dry weight losses due to insect infestation were recorded, however moisture evaporation caused weight losses between 6 and 8% during the period. Further investigations showed that careful handling and a dip concentration more in accordance with FAO/WHO MRL than the commercial practice will reduce the cost of insecticides from K.sh. 0.72 to K.sh. 0.23 per kg pyrethrum treated fish.