Isolation and recognition Saprolegnia species from fungal infected eggs of the rainbow trout in Mazandaran province farms

Fungal infection in the eggs of freshwater fish is well known as a problematic disease. That isolation and recognition Saprolegnia fungi from fungal infected eggs of the rainbow trout in Mazandaran province was the aim of this research. For this purpose fungal infected eggs were examined from six fish farm in the fall and winter 2005-2006. The eggs with fungi were inoculated on SDA, CMA, GPagar and hemp seed and sesame seed cultures in sterile tap water at room temperature (18-24°C). In this study recognized three genera and six species Saprolegniaceae members, based on morphological characteristics which contain: Saprolegnia, Achlya, Brevilegnia. Four species were identified in the genus Saprolegnia; S.mixta, S.parasitica, S.moniliphera, S.lapponica and one species was identified in the genus Achlya; A.oblongata. S.parasitica was isolated from almost all the farms. In addition, another nine genera and species were identified; Penicillium, Aspergillus, Paeciliomyces, Acremonium, Fusarium oxysporum, F.solani , Alternaria, Helminthosporium, Mucor.

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.

The study of histopathological damage in heart and bulbus arteriosus in rainbow trout

This study investigated the pathological changes of heart and bulbus artrius of rainbow trout breeders in several group of ages and density. The aim of study was to consider the process and the intensity of the heart and bulbus arteriosus damages in accordance to gender, age and stocking density of trout in three fish culture center (Zarghezel, Niyak in Haraz Region,Mazandaran and Espiran in Tabriz city environs). In field research, the all records the feed and feeding type, rate of mortality, stocking density of spawners and per spawners fishes, water chemical and physical specification was screened. Stocking density was considered as the most important stressor. 10 fish specimens from 7 weight groups (less than 90g, 90 to 300g, 300 to 500 g, 500 to 1000g, 1 to 3 kg, 3 to 5 kg, over 5 kg), totally 210 specimens were sampled and heart and bulbus arteriosus were taken. Samples were fixed in 10 % formalin and transferred to pathology laboratory of veterinary faculty of Tabriz Azad University. Histopathological slides and H&E staining were prepared from these samples. In total, 47 male and 73 female samples showed cardiovascular injury (29 cases in extensive system, 41 cases in semi intensive system, 50 cases in intensive system). The most important was damages, edema and hyperemia in spongy layer of atrium and ventricle muscles, but degeneration the muscle fibers, moderate edema , minor vascular damage. Hemorrhage as the effect of severs vascular damage, thrombus, sever inflammation, sever degeneration in muscle fiber, necrosis and fibrose were further pathological changed. The results of this study showed that the severity of damage increased by increasing the age (weight) of fishes. This situation was seen in all three culturing system (extensive system, semi intensive system, recirculation system). Histopathological changes is obviously seen in samples over 500g, therefore the damages were found to be important (P<0.05). Pathological effects and its severity in recirculation system was significantly high (P<0.05). Comparison with two other culturing system, histopathological changed in heart and bulbus arterius between male and female was significantly different.

Study on the inhibitory effects of native medicinal plants on growth and aflatoxin production by toxigenic Aspergilli isolated from rainbow trout feed

In the present study, natural occurrence of fungi and aflatoxin B1 (AFB1) in pellet feed and feed ingredients used for rainbow trout was investigated with emphasis to Aspergillus section Flavi members and medicinal plants inhibitory to Aspergillus growth and/or AF production. The feed samples were cultured on the standard isolation media including dichloran rosebengal chloramphenicol agar (DRCA) and Aspergillus flavus/parasiticus agar (AFPA) for 2 weeks at 28 °C. Identification of fungal isolates was implemented based on the macro- and microscopic morphological criteria. AFs were detected using high performance liquid chromatography (HPLC). Based on the results obtained, a total of 109 fungal isolates were identified of which Aspergillus was the prominent genus (57.0%), followed by Penicillium (12.84%), Absidia (11.01%) and Pseudallscheria (10.10%). The most frequent Aspergillus species was A. flavus (60.66%) isolated from all the feed ingredients as well as pellet feed. Among 37 A. flavus isolates, 19 (51.35%) were able to produce AFB1 on yeast extract-sucrose (YES) broth in the range of 10.2 to 612.8 [tg/g fungal dry weight. HPLC analyses of trout feed showed that pellet feed and all feed ingredients tested except gluten were contaminated with different levels of AFB1 in the range of 1.83 to 67.35 lig/kg. In order to finding natural inhibitors of fungal growth and/or AF production, essential oils (EOs) and extracts of 49 medicinal plants were studied against an aflatoxin-producing A. parasiticus using a microbioassay technique. The EOs was analyzed by gas chromatography/mass spectrometry (GC/MS). Based on the results obtained, Achillea millefolium sub sp. elborsensis, Ferula gummosa, Mentha spicata, Azadirachta indica, Conium maculatum and Artemisia dracunculus remarkably inhibited A. parasiticus growth without affecting AF production by the fungus. Besides of Thymus vulgaris and Citrus aurantifolia, the EO of Foeniculum vulgare significantly inhibited both fungal growth (-70.0%) and AFs B1 and G1 (-99.0%) production. The EO of Carum carvi and ethyl acetate extract of Platycladus orientalis suppressed AFs B1 and G1 by more than 90.0%, without any obvious effect on fungal growth. The IC50 values of bioactive plants for AFs B1 and G1 were determined in the ranges of 90.6 to 576.2 and 2.8 to 61.9 µg/ml, respectively. Overall, results of the present study indicate the importance of AF contamination of trout feed as a risk factor for fish farming and thus, an urgent necessity for constant monitoring of trout feed for any unacceptable levels of AF contamination. Likewise, antifungal activities of bioactive plants introduced here would be an important contribution to explain the use of these plants as effective antimicrobial candidates to protect feeds from toxigenic fungus growth and subsequent AF contamination.

Study of rainbow trout neoplasia in the region of Haraz

This study was carried out in order to determine neoplasia presence in rainbow trout in Haraz area. Neoplasia in various species of fishes including freshwater, brackish water and marine fishes was reported in other countries. But up to now there is no documented report of rainbow trout neoplasia in Iran. The study was performed in 20 farms of Haraz area during 2004-2005. All fishes of each farm firstly were observed in order to any abnormal mass which is suspicious to neoplasia. Besides in order to observation of external and internal organs, 20 fishes was sampled randomly from each farm and were examined clinically and necroptically. Any suspicious lesions were sent to pathology laboratory in 10% formalin followed by taking pictures of the lesions. Then histopathological evaluations were performed. From 400 fishes, 3 neoplastic cases including hepatocellular carcinoma along with bile duct papilloma, hepatocellular carcinoma and bile duct adenocarcinoma and intestine adenocarcinoma were found which all were in brooder fish.