Persistence of viable but non-culturable bacteria during the production and distribution of drinking water

The direct measurement of in situ respiring bacteria using 5-cyano-2,3-ditolyl tetrazolium chloride (CTC) shows that, especially for Gram-negative bacteria, large numbers of viable but non-culturable (VBNC) bacteria are present in finished water from a conventional water treatment plant, and the regrowth of bacteria along distribution networks can be seen rapidly by using this very sensitive technique. The level of bacterial inactivation with chlorine is much less important than has been previously supposed (based on experiments with non-injured laboratory strains of bacteria and classical culture techniques). Threshold values of VBNC bacteria leaving water treatment plants or regrowing along distribution systems have to be determined for better control of coliform regrowth and health- risks associated with the consumption of drinking water.

The diversity and ecological role of protozoa in fresh waters

Protozoa feed on and regulate the abundance of most types of aquatic microorganisms, and they are an integral part of all aquatic microbial food webs. Being so small, aerobic protozoa thrive at low oxygen tensions, where they feed (largely unaffected by metazoan grazing) on the abundance of other microorganisms. In anaerobic environments, they are the only phagotrophic organisms, and they live in unique symbiotic consortia with methanogens, sulphate reducers and non-sulphur purple bacteria. The number of extant species of protozoa may be quite modest (the global number of ciliate species is estimated at 3000), and most of them probably have cosmopolitan distributions. This will undoubtedly make it easier to carry out further tasks, e.g. understanding the role of protozoan species diversity in the natural environment.

Physico-chemical factors and bacteria in fish ponds

Analyses of pond water and mud samples show that nitrifying bacteria (including ammonifying bacteria, nitrite bacteria, nitrobacteria and denitrifying bacteria) are in general closely correlated with various physico-chemical factors, ammonifying bacteria are mainly correlated with dissolved oxygen; denitrifying bacteria are inversely correlated with phosphorus; nitrite bacteria are closely correlated with nitrites, nitrobacteria are inversely correlated with ammoniac nitrogen. The nitrifying bacteria are more closely correlated with heterotrophic bacteria. Nitrobacteria are inversely correlated with anaerobic heterotrophic bacteria. The correlation is quite weak between all the nitrite bacteria which indicates that the nitrite bacteria have a controlling and regulating function in water quality and there is no interdependence as each plays a role of its own. The paper also discusses how the superficial soil (pond mud down to 3.5 cm deep) and different layers of the mud affect the biomass of bacteria. The study shows that the top superficial layer (down to 1.5 cm deep) is the major area for decomposing and converting organic matter.

Heterotrophic marine bacteria as supplementary feed for larval Penaeus monodon

The findings are presented of a study conducted to use autochthonously obtained, nonpathogenic heterotrophic marine bacteria as a substitute feed for microalgae in rearing larval Penaeus monodon. Eleven strains were isolated: Micrococcus (MCC), Staphylococcus, Streptococcus, Bacillus (two strains; BAC-1, BAC-2), Pseudomonas (two strains; PSM-1, PSM-2), Vibrio parahemolyticus, V. fluviatilis, Moraxella (MOR) and Flavobacterium. Six nonhemolytic strains were then chosen for the Penaeus monodon larval feed trials: BAC-1, BAC-2, PSM-1, PSM-2, MCC and MOR. The study demonstrates that bacterial biomass could be further investigated as a partial substitute for microalgae in penaeid shrimp larval rearing.

Incidence of human pathogenic bacteria in shrimp feeds - a study from India

The incidence of various human pathogenic bacteria in commercially available and home-made shrimp feeds used on some farms in India was analyzed. The Total Heterotrophic Bacteria in the commercial feed samples ranged between 103–105 cfu g-1 and those in the farm-made feeds between 106-107 cfu g-1. No bacteria of significance to human health were found to be associated with any of the commercial feed samples analyzed, while farm-made feeds analyzed during the study showed a high incidence of various human pathogens such as Vibrio parahaemolyticus, V. cholerae, Escherichia coli and Staphylococcus aureus. Possible modes of contamination in feeds and ways to prevent them are discussed.

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.

Salt-tolerance in mangrove soil bacteria

A total of sixteen bacterial species were isolated from mangrove soils of Karachi, Pakistan. Twelve of the isolates were gram positive while four were gram negative. All sixteen species showed resistance to high concentration of streptomycin, however, resistance to chloramphenicol and tetracycline was variable. The isolates tolerated up to 110‰ salinity and accumulated sodium form the media.

Study on the intestinal bacteria of Labeo rohita (Ham.)

The quantitative and qualitative aspects of intestinal bacteria of rohu fish (Labeo rohita) showed that total viable count of bacteria ranged from 9.9 x 106 to 1.4 x 107 CFU/g of intestine in different age groups of fish. The bacterial load was highest in the month of July and lowest in January. The genera of the isolates from intestine included Coryneform, Micrococcus, Flavobacterium, Cytophaga, Achromobacter, Aeromonas Enterobacteriaceae and Vibrio. Coryneform was the dominant group throughout the study period followed by Micrococcus and Enterobacteriaceae. Marked variations in the bacterial load and generic composition of intestinal bacteria were evident during the study period in different age groups of rohu fish.

Effect of some environmental factors on the acute toxicity of deltamethrin to common carp: a laboratory study under aerobic condition

The toxicity of xenobiotic in aquatic ecosystems is influenced by many factors such as ambient temperature, water hardness, pond soil type, etc. In the present study, it was observed that air temperature, water hardness and soil sediment have profound influence on the toxicity of deltamethrin to common carp fry (ay. length 3.5 ± 0.5 cm, ay. weight 0.58 ± 0.25 g); 96h LC(sub)50 values for common carp at 38.07 ± 2.20°C maximum and 27.86 ± 1.22°C minimum air temperature in soft and very hard water were 0.102 and 0.495 µg lˉ¹, respectively. This value had increased significantly to 2.37 and 3.02 µg at 30.55 ± 1.21°C maximum and 26.04 ± 0.61°C minimum air temperature, respectively. When sediment was included, 96h LC(sub)50 at 38.07°C maximum temperature in very hard water was 1.808 µg 1ˉ¹ and this had increased to 8.073 µg 1ˉ¹ when tested at 30.55°C maximum temperature. Due to the 7.5°C increase in maximum and 1.7°C in minimum temperature, toxicity increased significantly. Lower toxicity in very hard water in comparison to soft water may be due to the lower solubility of deltarnethrin and high level of calcium. Adsorption reaction of deltamethrin with clay, humus, FeOOH, MnOOH and particulate organic carbon, and complexation reaction with dissolved organic carbon were responsible for the lowered toxicity in the experiment with sediment. Exposure time had no significant effect on acute toxicity of deltamethrin.

Marine bacteria as indicators of upwelling in the Sea

The “oxidase reaction” (using p-amino-dimethyl-aniline oxalate as the reagent) has been used to distinguish oxidase-negative from oxidase-positive bacteria from the sea, when grown on membrane filters. By this means, it has been shown (a) that under conditions of stable stratification of the sea as in the tropics, a relationship exists between the percentage incidence of oxidase negative bacteria in the flora and the depth of the water; (b) that the maximum value for this percentage incidence (100) is reached at or immediately below the upper limit of the oxygen minimum layer; (c) that this percentage value (expressed as Oxⁿvalues) may be used to demonstrate the movements of water masses during upwelling. Such upwelling as indicated by theoretical findings and by temperature determinations along two transects off the west coast of Ceylon during the north east monsoon, has been confirmed by the distribution of Oxⁿvalues at these transects.

Experimental pathogenesis of Aeromonas hydrophila bacteria in shing Heteropneustes fossilis(Bloch)

Pathogenicity of Aeromonas hydrophila bacteria was tested on the stinging catfish Heteropneustes fossilis. Before artificial infection the morphological, biochemical and physiological characters of Aeromonas hydrophila were studied. The infections were done by two different methods, viz., intramuscular (IM) and intraperitoneal (IP) injection. In infection experiment, each group of 10 fish were injected either intramuscularly or intra peritoneally with one dose higher than the LD50 dose (9.6 x 107 CFU/fish). All the fish tested died within 1 to 9 days. Both in cases of intramuscular and intraperitoneal injection, external pathology were found. Haemorrhagic lesions were evident at the site of injection. The posterior end of the body surface was found to develop greyish-white lesion that was extended up to caudal fin. Hyperemic anal region and the fin bases were also observed. Total bacterial loads in liver, kidney and intestine were determined. Aeromonas hydrophila could be isolated from liver, kidney and intestine of the experimentally infected fish. In case of intramuscular injection the highest and the lowest bacterial load was found to be 2.4 x 107 CFU/g of liver and 2.1 x 102 CFU/g of kidney and in case of intraperitoneal injection they were found to be 3.6 x 106 CFU/g of kidney and 1.2 x 104 CFU/g of kidney respectively. It was concluded that A. hydrophila could cause serious disease condition to Heteropneustes fossilis and its pathogenesis in the fish was also very efficient.

Distribution of drug resistant luminous bacteria in penaeid shrimp grow-out ponds

Distribution of luminous bacteria (LB) in penaeid shrimp grow-out pond water in semiintensive seawater farming system and their resistance to 15 antibacterials were investigated. Total viable counts and luminous bacterial counts in pond water ranged from 2.00xl03 to 1.35xl04/ml and l.OOxl01 to 8.00Xl02/ml, respectively. The percentage composition of LB in the total viable population increased significantly with period of culture. Five species of LB such as Vibrio fischeri, V. harveyi, V. orientalis, V. splendidus 1 and Photobacterium leiognathi were encountered. V. harveyi was the dominant species, constituting >80% of the total LB. Multiple antibiotic resistance was more common in these LB. Pond water isolates showed resistance to at least four antibacterial agents.

Effect of delayed icing on quality changes and shelf-life of some freshwater fish from Bangladesh

The quality and shelf-life of three freshwater fish species of Bangladesh, catla (Catla catla), magur (Clarias batrachus) and tilapia (Oreochromis niloticus) stored at room temperature and ice were evaluated. Live fishes were killed by cranial spiking and stored at room temperature (27-30 °C), ice stored immediately after death, 5 hr after death and 10 hr after death. The shelf-life and quality of the fishes were evaluated by organoleptic method, rigor-mortis studies and bacteriological assessment. Fishes kept at room temperature became organoleptically unacceptable within 16-20 hr duration. Ice stored fishes showed considerable differences in their shelf-life when icing was delayed for different duration. Shelf-life of catla ice stored immediately after death was 20 days but shelf-life reduced to 12 days when icing delayed for 10 hr after death. Similar trend was observed for two other fish species magur and tilapia. Rigor-index of the fishes stored under different conditions also considerably varied among the three fish species, particularly effect of delayed icing was very much evident. Bacteriological study showed patterns of Aerobic Plate Count (APC) at the end of shelflife study when fishes became organoleptically unacceptable were more or less similar for all the three fish species stored under different conditions. No definite pattern was observed in the generic distribution of bacteria in different fish species under different storage conditions. Micrococcus, Coryneforms, Pseudomonas and Achromobacter were the dominant groups of bacteria isolated from the fishes spoiled at room temperature and ice stored condition.

Inhibition of seafood-borne bacteria in cooked mackerel (Rastrelliger kanagurta) fish meat by lactic acid bacteria

Antagonistic activity of lactic acid bacteria (LAB) namely Streptococcus faecalis, Pediococcus cerevisiae and Lactobacillus casei was tested against seafood-borne bacteria such as Staphylococcus aureus, Bacillus cereus, Escherichia coli, Clostridium perfringens and Listeria monocytogenes. Three lactic acid bacteria such as Streptococcus faecalis, Lactobacillus casei and Pediococcus cerevisiae were coated on cooked mackerel meat, individually and in combination against fish-borne bacteria. S. faecalis inhibited C. perfringens in individual coat by 3.7 log units as compared to control, whereas L. casei did not inhibit C. perfringens. P. cerevisiae inhibited S. aureus by 5 log units. L. casei, inhibited L. monocytogenes by 3.3 log units on the third day of storage as compared to control. On the other hand, S. aureus and B. cereus were inhibited on the third and second day by 4.9 log and 5.2 log units respectively. B. cereus, S. aureus, L. monocytogenes were the most sensitive to all three LAB. C. perfringens was the least inhibited among all the seafood-borne bacteria tried. Multiple LAB or LAB strains in combination showed much earlier inhibitory activity on seafood-borne bacteria than single LAB coat.

Improved handling and preservation of freshwater giant prawn, Macrobrachium rosenbergii for producing safe and wholesome products

Studies were undertaken to evaluate the quality changes in freshwater giant prawn, Macrobrachium rosenbergii during various storage conditions of handling and preservation and producing safe and quality products. The samples kept in ice immediately after catch with head-on and head-less condition were found to be acceptable for 6 days and 7 days, respectively. Delaying of icing considerably shortened the shelf-life. The pH value increased from 6.36 to 8.0 after 10 days in ice. The initial average TVB-N value of sample increased from below 10 mg/100 g to 25 mg/100 g with the lapse of storage period. The Ca++ ATPase activity in presence of 0.1M KCl slightly decreased at the end of 10 days of ice storage. Immediately after harvest, initial aerobic plate count (APC) was 2.88x10^6 CFU/g which gradually increased to 1.12x10^8 CFU/g after 6 days in ice storage and showed early signs of spoilage. Initial bacterial genera in the prawn iced at 0 hours were comprised of Coryneform (22.21 %), Bacillus (7.40%), Micrococcus (11.11 %), Achromobacter (48.14%), Flavobacterium/Cytophaga (7.40%), Pseudomonas (3.70%) and Aeromonas (3.70%). During ice storage Coryneforms and Bacillus were always dominating along with less prominent ones - Micrococcus, Achromobacter and Flavobacterium. Studies were conducted on the stability of myofibrillar protein of M. rosenbergii under different storage and pH conditions. The influence of a wide range of pH on the remaining Ca++ ATPase activity of M. rosenbergii muscle myofibrils after storage at -20°C for 2 days, at 0°C for 2 days and at 35°C for 30 minutes demonstrated that ATPase activities were lower in acidic and alkaline pH regions and the activity remained relatively high. Mg++ ATPase activities both in presence and absence of Ca++ remained high at neutral pH compared to those of acidic and alkaline region. The solubility of myofibrillar protein decreased gradually both in acidic and alkaline pH regions. The study also examined the bacteriological quality of freshly harvested M. rosenbergii, pond sediment and pond water from four commercial freshwater prawn farms at Fulpur and Tarakanda upazilas in the district of Mymensingh. The study included aerobic plate count (APC), total coliform count, detection, isolation and identification of suspected public health hazard bacteria and their seasonal variation, salt tolerance test, antibiotic sensitivity test of the isolates and washing effect of chlorinated water on the bacterial load in the prawn samples. APC in sediment soil and water of the farm and gill and hepatopancreas of freshly harvested prawns varied considerably among the farms and between summer and winter season. The range of coliform count in water, gill and hepatopancreas ranged between 6 - 2.8x10^2 CFU/ml, 1.2x10^2 - 3.32x10^2 CFU/g and 1.43x10^2 - 3.89 x10^3 CFU/g, respectively. No coliform was detected in pond sediment sample. Suspected health hazard bacteria isolated and identified from pond sediment, water, gill and hepatopancreas included Streptococcus, Bacillus, Escherichia coli, Klebsialla, Salmonella, Staphylococcus, Pseudomonas and Aeromonas. Bacillus, Salmonella and Staphyloccus [sic], and were found to be highly salt tolerant and capable of growing at 10% NaCl. The antibiotic discs with different concentration of antibiotics were used for the sensitivity test. The organisms were found to be most sensitive against Tetracyclin and Gentamycin.