Identification, diversity and distribution patterns of fish larvae in Khark & Kharko Coralline Islands ecosystem – Persian Gulf, by GIS method

Khark & Kharko Islands are the last Northern point for fringing coral reefs in Iranian side of the Persian Gulf. These Coralline habitats are the Protected Area and Wildlife Refugees with the total area of 2400 ha which located in the territory of Bushehr Province. This research carried out during 2006-2007 with monthly sampling from 12 stations, which selected around Islands and inshore waters with maximum depth of 20 meter. Sampling was conducted using by Bongo-Net plankton sampler with 500μ of mesh size. Totally, 1808 specimen from 45 family fish larvae was identified in studied area, including: 21 coralline fish larva families and 24 shore fish larvae such as pelagic and demersal fishes which some of them known as indicator, sentinel or endemic species for coral reef ecosystems. The results was shown that coral reef diversity in coral reefs (Khark & Kharko Islands) is more than other habitats such as estuary and river mouth, creeks, mangrove forest sites, and off shore water of the Persian Gulf and Oman Sea Iranian side. Among Identified families, Clupeidae, Blenniidae, Sillaginidae, Atherinidae and Tripterygiidae; with more abundance were dominant families in studied area. The pick of fish larvae abundance family were estimated in spring. There were significant differences between seasonally abundance and sub areas, but there were not significant differences in diversity indexes between Khark and Kharko stations with coastal stations (p< 0.05). The mean abundance of fish larvae were estimated 18.7083 larvae under 10m² of sea surface, and the mean diversity indexes and evenness were estimated 0.7135 and 0.565342 consequently, that was showed the area is under ecological stress for fish larvae, and wasn’t stable. Therefore, from the ecological point of view, only some of the fish larvae groups as like Clupeidae were dominant. Thus, they were the main cause of the fish larvae abundance change in studied area. Due to geographical location of Khark and Kharko Islands and among the environmental parameters, Its seems that the condition of sea current is the main cause for present or absent and distribution patterns of fish larvae in area. Abundance of fish larvae in west of Islands was higher than eastern parts in the spring. But this condition will be reversed in eastern part of Island and several coastal stations, so that the Islands surrounding clock wise current to cause fish larvae distribution patterns.

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.