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.

Histological distribution and ultrastructure of exocrine pancreas in Indian major carp (Labeo rohita Ham.) and its alteration in aflatoxicosis

The distribution pattern of exocrine pancreas in Labeo rohita besides its general location along the course of intestinal mesentery was studied. It is evenly distributed within the liver around portal vessels and also within the spleen near a blood vessel. On ultrastructure, two cell types of different degrees of staining intensities containing abundant rough endoplasmic reticulum, mitochondria, pre-zymogen and zymogen granules were marked. During aflatoxicosis, the mesenteric pancreas and hepatic pancreas were mostly affected revealing necrotic changes to acini. The zymogen granular activities were markedly reduced. Ultra structurally, the rough endoplasmic reticulum was fully dilated and formed whorled pattern. The damage to the exocrine pancreas might be affecting digestive enzymes' secretion which may be one of the causes of aflatoxin-induced anorexia in fish.

Study of distribution, growth and reproduction polychaetes Nereidae in intertidal zone of Bandar Abbas and effect dominant species on white Indian shrimp brood stocks (Fenneropenaeus indicus)

Distribution of polychaetes worms Nereidae in intertidal zone of Bandar Abbass indicated that distribution rate of worms from the west to east for the reason status of seabed type, habitat and feeding substrate used to worms increased. Also investigation description that last-mentioned worms are belong to Nereididae family, Perinereis genus and species of Perinereis nuntia. By viewpoint seabed soil types experiments implement indicated Nereididae worms have higher survival with sandy clay loam soil texture. Statistical analysis showed positive relation coefficient correlation of Pearson between substrate type to frequency of worms and too substrate type with soil organic matter value (P<0.05). By viewpoint feeding also experiments with different feed treatment indicated that Nereid worms is Omnivore but in natural condition more utilized algae substrate special Entermorpha and survival rate had the worms last-mentioned in dietary treatments on this algae 93/3% different significant with other treatments (P<0.05). In this project pond-reared white Indian shrimp (Fenneropenaeus indicus) were used in four maturation and spawning trials lasting 60 days. one group of shrimps was fed a formulated pelleted special of broodstock diet only, a second group was fed squid meat diet; a third group of shrimps with mollusca (Solen vagina) meat diet and fourth group was fed polychaete worms Nereidae family, collected in intertidal zone of Bandar Abbass. Four types of dietary treatments (M1, M4) were given to separate batches that were run in three duplicate. Results of experiments demonstrate that more grew rate and maturation and spawning rate and we found that shrimps fed with polychaete worms have the best condition and then shrimps fed with solen. This is demonstrate furthermore environmental circumstance, endocrine hormone, types of feed important of maturation and spawning of shrimps and in polychaete worms used HUFA help to maturation ovarian in shrimp.

Supply of Astaxanthin and its combinations through live feed (Moina micrura) enrichment affects the growth, survival and fatty acid profile of Macrobrachium rosenbergii larvae

A study was carried out with three replicates to determine the effects of feeding Moina micrura enriched with astaxanthin alone (M1) or astaxanthin in combination with either vitamin E (M2), vitamin D (M3) or Cod Liver oil (M4) on the growth, survival and fatty acid composition of giant fresh water prawn Macrobrachium rosenbergii (de Man) larvae. Growth rate was expressed as the time taken to the settlement of 95% post larvae. Maximum growth, the lowest time taken to the 95% PL settlement (38.5±0.50 days), was observed in larvae fed with M3 Moina. The highest survival rate (66.0±1.00%) was observed in those fed with M4 Moina and the second highest survival (61.0±1.00%) and growth rates (40.0±0.00 days) were shown with M2 Moina. The minimum values for both growth (42.5±0.50 days) and survival (33.0±1.50%) were observed in the group fed un-enriched Moina. Results also showed that the survival of prawn larvae increased as the quantities of eicosapentaenoic acid (EPA) and docosahexaenoic acid (DHA) increased in the dietary Moina. The highest levels of EPA (5.57±0.21%), DHA (3.50±0.21%) and highest total Highly Unsaturated Fatty Acids (HUFA) (13.87±0.68%) were seen in the Moina fed on astaxanthin and Cod Liver Oil (CLO). The results of the study showed that the nutritive quality of Moina, with respect to important fatty acids, can be increased by enrichment and will influence the growth, survival and the fatty acid composition of fresh water prawn larvae fed on them.

Isolatin [sic] the bacteria produced biactive [sic] metabolits [sic] cytotoxic compound in order to prduction [sic] anticancer substance in the coral regin [sic] the Persian Gulf (Larak Island)

In the present investigation the marine bacteria isolated from corals, sponges sea water and sediments of coral regions in the larak Island located in the Persian Gulf and were examined for ability to produce cytotoxic metabolits in order to use as an anticancer compounds. Cytotoxic effect were isolated bacteria from different samples and were examined by Artemia Cytotoxic Bioassay test, in which 4.5 percent of sea waters, 12 percent of sediments and 28 percent of marine invertebrat showed cytotoxic activity, using Brine Shrimp Bioassay test. Streptomyces S-2004 isolated from soft coral specified as Sinularia erecta had LC50=0.5mg/m1 in Brine Shrimp Bioaassay test. The streptomyces S-2004 produced cytotoxic metabolits in low nutrient condition and sea water medium after 7 days on 250 rpm shaken in vitro condition. The extract partially were semipurified. Then ethyl acetate extraction from aceton extracted of bacterial plate had cytotoxic effect (LC50=4.19ktg/m1) in Human epidermoid carcinoma of mouth cells (KB) by using neutral red assay. Morphological effects of this extract on KB cells showed turgescence, cellular blebs and apoptosis which was a proof for anticancer compounds of the extract. It is seems that streptomyces S-2004 is a new strain and could be introduced as a talented bacteria, which produced cytotoxic metabolits.