Microbial community analysis of Acropora palamata mucus swabs, water and sediment samples from Hawksnest Bay, St. John, U.S. Virgin Islands

Colonies of the scleractinian coral Acropora palmata, listed as threatened under the US Endangered Species Act in 2006, have been monitored in Hawksnest Bay, within Virgin Islands National Park, St. John, from 2004 through 2010 by scientists with the US Geological Survey, National Park Service, and the University of the Virgin Islands. The focus has been on documenting the prevalence of disease, including white band, white pox (also called patchy necrosis and white patches), and unidentified diseases (Rogers et al., 2008; Muller et al., 2008). In an effort to learn more about the pathologies that might be involved with the diseases that were observed, samples were collected from apparently healthy and diseased colonies in July 2009 for analysis. Two different microbial assays were performed on Epicentre Biotechnologies DNA swabs containing A. palmata coral mucus, and on water and sediment samples collected in Hawksnest Bay. Both assays are based on polymerase chain reaction (PCR) amplification of portions of the small rRNA gene (16S). The objectives were to determine 1) if known coral bacterial pathogens Serratia marcescens (Acroporid Serratiosis), Vibrio coralliilyticus (temperature-dependent bleaching, White Syndrome), Vibrio shiloi (bleaching, necrosis), and Aurantimonas coralicida (White Plague Type II) were present in any samples, and 2) if there were any differences in microbial community profiles of each healthy, unaffected or diseased coral mucus swab. In addition to coral mucus, water and sediment samples were included to show ambient microbial populations. In the first test, PCR was used to separately amplify the unique and diagnostic region of the 16S rRNA gene for each of the coral pathogens being screened. Each pathogen test was designed so that an amplified DNA fragment could be seen only if the specific pathogen was present in a sample. A positive result was indicated by bands of DNA of the appropriate size on an agarose gel, which separates DNA fragments based on the size of the molecule. DNA from pure cultures of each of the pathogens was used as a positive control for each assay.

Influence of microbial interactions on the susceptibility of Karenia spp. to algicidal bacteria

A bacterial strain (D38BY) belonging to the family Flavobacteriaceae and antagonistic towards an algicidal bacterium (strain S03; Flavobacteriaceae) was isolated from a culture of the red tide dinoflagellate Karenia brevis that had previously been characterized as resistant to attack by strain S03. This antagonistic bacterium increased the survival time of otherwise susceptible, bacteriafree K. brevis cultures in a concentration-dependent manner during exposure to the algicidal bacterium. Experimental evidence indicated that direct contact was required in order for strain D38BY to inhibit the killing activity of algicidal strain S03. While further work is needed to determine its precise mode of action, the antagonistic properties of strain D38BY provide further evidence that the resistance or susceptibility of certain algal taxa to algicidal attack can be more a function of interactions within the ambient microbial community than an intrinsic property of the alga.

The coastal environment and human health: microbial indicators, pathogens, sentinels and reservoirs

Innovative research relating oceans and human health is advancing our understanding of disease-causing organisms in coastal ecosystems. Novel techniques are elucidating the loading, transport and fate of pathogens in coastal ecosystems, and identifying sources of contamination. This research is facilitating improved risk assessments for seafood consumers and those who use the oceans for recreation. A number of challenges still remain and define future directions of research and public policy. Sample processing and molecular detection techniques need to be advanced to allow rapid and specific identification of microbes of public health concern from complex environmental samples. Water quality standards need to be updated to more accurately reflect health risks and to provide managers with improved tools for decision-making. Greater discrimination of virulent versus harmless microbes is needed to identify environmental reservoirs of pathogens and factors leading to human infections. Investigations must include examination of microbial community dynamics that may be important from a human health perspective. Further research is needed to evaluate the ecology of non-enteric water-transmitted diseases. Sentinels should also be established and monitored, providing early warning of dangers to ecosystem health. Taken together, this effort will provide more reliable information about public health risks associated with beaches and seafood consumption, and how human activities can affect their exposure to disease-causing organisms from the oceans.

Studies of microbial abundances in mangrove habitat along the Karachi coast

Three species of bacteria, 8 species of fungi and 3 species of VAM-fungi were isolated from the soil substrata supporting Avicennia marina which comprises the majority of mangrove vegetation along the Karachi coast. The species abundances for fungi and bacteria were greater at one site (Sandspit) supporting healthy mangrove growth with soil pH 7.8, EC 16.2mmhos/cm², TSS 2.57% and available phosphorus 0.008% than at the other site (Korangi creek) with stunted growth of mangrove where the soil samples showed pH 7.9, EC 18.8mmhos/cm², TSS 1.45% and available phosphorus 0.001%. Symbiotic association by vasicular-arbuscular mycorrhizal (VAM) fungi in the roots of mangrove plants was also observed on a small scale at Korangi creek where the substratum was undergoing microbial degradation.

Ensiled product from fish by microbial fermentation

Fish ensilage for animal feed stuff was prepared from Jew fish (Pseudosciaena spp.) and Silver bellies (Leiognathus spp.) by fermentation with pure culture of Lactobacillus piantarum NCIB 6105. The precooked ensiled product gave better product of fish silage (high content of lactic acid, about 5%). Protein Nitrogen content ranged between 1.76 to 1.94%. During storage for one year, the Protein Nitrogen loss was not significant. The material can be used as a supplemental animal ration.

Studies on the possible sources of microbial contaminations of processed fishery products

An attempt has been made in the present study to estimate and describe in detail the nature and extent of contamination of processed fishery products. In large scale prawn processing when the preprocess preparation is elaborate, the industry in India has found it advantageous to establish the primary processing centers away from the processing factories. The data collected have clearly indicated that if such processing centers are not properly organized there is a possibility of greater contamination of the products at this stage. The data collected during the course of this investigation have given the basis for the measures to be taken for the maintenance of bacterial quality of prawn during different stages of processing.

Comparison of growth and conversion efficiency of the freshwater prawn Macrobrachium malcolmsonii (H. Milne Edwards) juveniles fed with formulated and commercial feeds

A nutritional study was carried out to know the feasibility of formulated and commercial feeds on the growth of juveniles of the freshwater prawn Macrobrachium malcolmsonii. Best relative growth rate was observed for feed 1 followed by feeds 2 and 3. Animals fed with feed 1 showed higher production than the other feeds. Higher assimilation efficiency was noticed in the animals provided with feeds 5 and 4. Higher gross and net growth efficiencies were observed in the animals offered feed 1. Among the commercial feeds, feed 3 may considered to be suitable alternative to feed 1.

The effect of feeding rate on the growth, food conversion and protein efficiency of silver dollar, Metynnis schreitmulleri (AHL), fry

A 30-day experiment was conducted to determine the effects of varying feeding rates on the growth of fry of silver dollar, Metynnis schreitmulleri (Ahl). Silver dollar fry with an average initial body weight of 1.100 ± 0.029 g were collected from a local fish breeder and fed a diet (35% protein and 6% fat) at the rate of 3, 6 and 9% of body weight per day in two equal meals. Per cent weight gain increased from 54.54 to 118.18 with increased feeding rates, which were significantly different (p<0.05) from each other. The highest specific growth rate was obtained in the fry fed at 9% body weight per day. In another study for 30 days, the effects of feeding frequency on growth, feed conversion and protein efficiency of silver dollar fry were evaluated. Groups of silver dollar fry with an initial individual weight of 0.700 ± 0.019 g were offered feed continuously for 24 hours during the day or night at different time intervals with varying meal sizes. The night time feeding with two meals gave the lowest weight and length gains (0.985 g and 0.30 cm). The growth rates of fish fed during day with three equal-sized meals are significantly (p<0.05) higher (4.66%) than the other treatments.

Effects of gamma radiation on nutritional and microbial quality of Pampus chinensis (Euphrasen 1788)

To evaluate the efficiency of gamma radiation in combination with low temperature Chinese pomfret, Pampus chinensis were preserved by the treatment of different doses of gamma radiation (3, 5 and 8 KGy) at freezing temperature (-20°C) during 90 days of storage period. Quality assessments for fish were carried out at an interval of 15 days during the storage period. Quality assessments were done by organoleptic, chemical (Total Volatile Nitrogen, TVN and Trimethylamine, TMA) and microbiological (Total Bacterial Count, TBC and Total Mould Count, TMC) evaluation. From the analysis of all parameters, maximum shelf-life was observed for irradiated (8 KGy) sample. It remained acceptable up to 75 days and that was the highest duration among 4 types of samples.

Effect of microbial biofilm in the nursery phase of mrigal, Cirrhinus mrigala

The experiment was conducted for 35 days in nine cement tubs (1 x 1 x 1 m) having 15 cm sandy-loam soil base with three treatments in triplicate, viz., cow dung alone at the rate of 1 kg/tub (T sub(1)), cow dung at 1 kg/tub and feed at 10% body wt/d in two meals (T sub(2)), and cow dung at 1 kg and paddy straw at 200 g/tub (T sub(3)). Both manure and substrate were added on dry weight basis. All the tubs were stocked with 10 fry each mrigal (100,000/ha) of average weight of 0.09 g, seven days after the addition of manure and substrate. The total plate count of bacteria in water did not vary much between the treatments and the mean values were 5.13, 5.49 and 5.85 (CFU x 10 super(4)/ml) in T sub(1) T sub(2) and T sub(3) respectively. The number of phytoplankters and zooplankters in water differed significantly between the treatments. The average number of attached algae (no./cm³) and fish food organisms (no./cm³) recorded on the substrate were 145.28 and 70.67, respectively. The mean final weight of mrigal differed significantly (P < 0.05) between the treatments with T sub(3) registering the highest value of 6.93 g followed by T sub(2) (5.01 g) and T sub(1) (3.37 g). The specific growth rate and growth increment of fish also followed the same trend as that of weight recorded in the different treatments. Survival was higher in T sub(2) (83.33%), followed by T sub(3) (80.00%) and T sub(1) (76.67%). The study demonstrates that by the introduction of biodegradable substrates like paddy straw into the culture systems, significantly higher growth and survival can be obtained in the nursery rearing of mrigal.

Utilization of prawn waste: isolation of chitin and its conversion to chitosan

process is described for the preparation of chitosan from prawn waste. The process involves extraction of protein using 0.5% sodium hydroxide solution, bleaching the protein free mass with bleach liquor containing 0.3-0.5% available chlorine followed by demineralisation with 1.25 N hydrochloric acid in the cold and deacetylation using 1:1 (w/w) sodium hydroxide solution at 100°C for 2 hours.

Response of extract release volume to the variations in pH and microbial load during spoilage of fish at refrigeration temperature

This report briefly describes the microbial status and storage properties of fish raised under composite fish culture in sewage fed ponds.

Comparative effect of Pediococcus acidilactici and Lactococcus lactis on growth performance, survival and enzyme activity of western white leg shrimp (Litopenaeus vannamei)

This study was done in Shahid Kiani Marine Aquaculture Development Center, Choebde, Abadan in order to evaluate the effects of Pediiococcus acidilactici, Lactococcus lactis and vitamin C on growth performance, survival, enzymatic activities and immune responses of L. vannamei during three months. Treatments were included control group, Pediiococcus and Lactococcus treatments which fed with diet containing 1×10P9P cfu gP_1P bacteria and vitamin C. At the end of the experiment, the growth factors, immune parameters, digestive enzymes, intestinal, histology of intestine, carcasses and microbial flora (bacterial total count and lactic acid count) were evaluated. The results indicated that administration of lactobacillus had significant effects on the growth factors as the highest weight, increase specific growth rate, relative growth rate, feed conversion ratio and protein efficiency in the shrimps received pediococcus and then Lactococcus (P<0.05). The best immune function was also observed in the shrimps fed by probiotics, so that proteins and hemoglobin̛ hemolymph, phenoloxidase activity and challenged with V. parahaemolyticus showed a statistical difference comparing to the control group and the group received vitamin C (P<0.05). Some digestive enzymes, in pediococcus treatment showed a significant increase when compared to other treatments (P<0.05). Significant changes in bacterial intestinal flora were observed in probiotic groups compared with control and vitamin C groups (P < 0.05). Histological results showed the positive effects of probiotics in the gut (P < 0.05). While these supplements cannot caused to significant impacts on the shrimp carcass composition (P ˃ 0.05). As a result pediococcus group had the best performance among treatments.

Study of physicochemical, microbial and sensory properties of surimi produced from black mouth croaker (Atrobucca nibe) at freezing condition (-18ºC)

Black mouth croaker (Atrobucca nibe) is considered as a new valuable fish stock in the Oman Sea. In this study, surimi was manufactured from nonmarket size of the fish, manually and different cryoprotectant agents were added to the surimi. Finally changes in physiochemical, microbiological and sensory quality, characteristics of the surimi and kamaboko gel samples were assessed during 6 months at freezing storage (-18ºC). Surimi samples with the addition of Iranian tragacanth gum (TG), xanthan gum (XG), chitosan (CS) and whey protein concentrate (WPC) at 1% (w/w) were prepared to evaluate their impacts as a cryoprotectant on the surimi, individually. The results showed that the whiteness and lightness indexes in all surimi samples were gradually decreased during frozen storage. This trend of decreasing was more intensity in the control sample from 61.08±0.131 to 54.21±0.067 was recorded (p<0.05). Water holding capacity (WHC) in all treatments was decreased during 6 months. The lowest WHC (g/g) was obtained in the surimi without cryoprotectants and maximum WHC was measured in Tcs and Twpc samples, respectively (p<0.05). The lowest breaking force was calculated in Txg (166.00±22.627 g) and Tc (271.50±263.16 g) during 6 months at frozen storage, respectively (p<0.05), while Twpc treatment with slight variations showed the highest breaking force (p<0.05). Also, the lowest gel strength was obtained in Txg (68.22±6.740 g.cm) after 6 month of frozen storage (p<0.05). All Kamaboko surimi gels texture profile analysis parameters decreaced with increasing shelf life. This decreasing trend in the control sample was more severe. Floding results were reduced in all samples during storage (p<0.05). The best protective results probably were obtained in WPC, chitosan and commercial cryoprotectant agents, respectively due to protein stabilization of myofibrillar proteins and the protein-protein network structure, leading to the formation of surimi gel with strong textural properties during frozen conditions. The average number of surimi polygonal structures were significantly decreased (number per mm2) and their area were significantly increased (μm2) in all treatments (p<0.05). With increasing storage time, moisture, protein contents and pH were decreaced. Maximun TVB-N index was calculated in Tc (7.93±0.400 mg/100g) and Txg (7.88±0.477), respectively (p<0.05). TBRAs index was increased in all treatments during frozen storage, while this trend was reached in maximum value in Tc (p<0.05). Sensory evaluation of the fish finger quality characteristics (color, odor, texture and overall acceptability) preapare from frozen black mouth croaker surimi was decreaced during 6 month frozen storage. After the period of frozen storage the highest quality scores were measured in Twpc, Tcs and Tcc samples, respectively (p<0.05). In this study, coliform bacteria were not found in all treatments during frozen storage. The surimi sample containing chitosan showed lower mesophilic and psychrotropic bacteria (log cfu/g) than other treatments during frozen storage (p<0.05). Salt-soluble proteins extractions of all treatments were decreased during frozen storage. This decreacing trend was highest in Tcs (45.74±0.176%) and lowest in Tc treatments after 6 month of frozen storage (29.92±0.224%) (p<0.05). Although commercial cryoprotectant agents were successful in limiting the denaturation of proteins but sugar contents were not accepted for diabetics or those who disagree with the sweet taste and high calorie food. Hence, commercial cryoprotectant agents can be replaced with whey protein concentrate and chitosan at 1% level (w/w) consider that they were showed proper protection of the surimi myofibrillar proteins during storage.

Study on effects of extracted probiotics from microbial flora of digestive system on growth indices in beluga (Huso huso) and Persian sturgeon (Acipenser persicus)

Isolation of Lactic Acid Bacteria (LAB) was carried out from gastro intestinal tract of beluga and Persian sturgeon at international sturgeon research institute and PCR has been used for bacteria Identification. Two species of LAB including Enterococcus seriolicida and Leuconostoc mesenteroides were isolated from Gastrointestinal tract (GI) of persian sturgeon in this study and the counts of Leu. mesenteroides (4.63×102 CFU/gr of GI) was significantly higher than other species. Lactobacillus curvatus, Lactococcus raffinolactis, Lactococcus lactis and Streptococcus sp. were also isolated from GI of beluga and maximum counts was belonged to Lb. curvatus (4.63×102 CFU/gr of GI) in this species. Dominant species were lyophilized and adding to the water since start of mix feeding of sturgeon with different counts including 2×109, 5×109 and 9×109 CFU/gr of live food, 4 times a day. The results revealed that the maximum and minimum growth rate and protease, amylase, and lipase activity in beluga was gained by using of Lb. curvatus with total viable count of 9×10 9 CFU/gr of live food and Leu. mesenteroides with total viable count of 9×109 CFU/gr of live food. According to the results of this study, the maximum and minimum growth rate and protease, amylase, and lipase activity in Persian sturgeon was gained by using of Leu. mesenteroides with total viable count of 2×10 9 CFU/gr of live food and Lb. curvatus with total viable count of 9×109 CFU/gr of live food. Histological study showed that gastrointestinal development was same during larva rearing in control and other treatments but the size of liver was bigger in treatments that received nonspecific LAB in both species. According to the results, positive effects of using dominant specific LAB bacteria for larviculture of sturgeon has been proved in this study.