Studies on the chemical control of psychrophilic bacterial spoilage of fish. 2 - The effect of antibiotics on the growth of psycrophilic bacteria isolated from marine fish

Among the various antibiotics tried, tetracyclines particularly chlorotetracycline (CTC), chloramphenicol and chlorostrep were found to be fairly effective at 8 and 10 p.p.m. levels. The order of sensitivity to CTC among the six genera studied was found to be Achromobacterbacteriumbacterium and Pseudomonas were generally more resistant to antibiotics than species belonging to other genera. Among the resistant organisms, Achromobacter superficialis was found to be most resistant and only CTC at 10 p.p.m. level could partially inhibit this culture. The useful combinations of antibiotics for commercial application are discussed.

Studies on the chemical control of psychrophilic bacterial spoilage of fish. 4 - The effect of chemical preservatives on the growth of psychrophilic bacteria isolated from marine fish

Dehydroacetic acid and ammonia were found to be very effective in checking the growth of all the cultures at all concentrations tried. The two nitrofuran derivatives namely, semicarbazone and AF-2 were fairly effective, semicarbazone being more effective than AF-2. Sodium nitrite was found to be totally ineffective against the cultures at all concentrations tried.

Chemical control of psychrophilic bacterial spoilage of fish 1. Isolation and identification of psychrophilic bacteria from marine fish

Making use of the streak plate technique and low temperature incubation, 28 cultures belonging to six genera namely, Achromobacter, Flavobacterium, Pseudomonas, Micrococcus, Vibrio and Alcaligenes were isolated from different varieties of marine fish. The growth studies indicated that all of them were able to grow between -5 and +5°C within a week's time and none of them showed growth at 37°C. The optimum temperature of growth for all these cultures was in the range 25-28°C. Among these only one, i.e., a Vibrio sp., was found to be an obligate psychrophile.

Influence of a fish bacterium Lactobacillus sp. on the production of swordtail xiphophorus helleri (Heckel 1848)

The influence of a fish gut bacterium Lactobacillus sp on the production of swordtail Xiphophorus helleri was studied for a period of one year. The Lactobacillus sp P21 produced bacteriocin-like inhibitory substance and exhibited wide spectrum of action against Aeromonas hydrophila, Bacillus spp, Pseudomonas spp and Citrobacter freundi in vitro. The growth performance of X. helleri reared in the presence of Lactobacillus P21 at 106/ml rearing water was better than the control. The total plate counts, total MRS agar counts and the counts of motile aeromonads, presumptive pseudomonads, lactose fermenters and lactose non-fermenters in the gut of probiotic group were comparatively low than the control. On day 60 the count of Lactobacillus sp P21 was observed to be log 5.28/g in the gut of X. helleri indicating colonization of this bacterium in the gastrointestinal tract. The fecundity of X. helleri was in the range of 9-134. On average, it produced from 39.42±18.72 fry/female in control group to 53.00±23.57 fry/female in probiotic group. The increase in average fecundity in probiotic group over the control group was about 25%. There existed significant difference between probiotic group and control in respect of average fecundity/female (p<0.02), average number of fry survived /female (p<0.006) and average number of fry dead/female (p<0.029). The results of the present study demonstrated that the rearing of X. helleri in probiotic-enriched water have growth inducing ability and favourably influenced the reproductive performance in terms of high fecundity, high fry survival, reduced fry mortality and reduced fry deformity.

Antibacterial activity of a marine bacterium against pathogenic and environmental isolates of vibrio species

The marine environment covers three quarters of the surface of the planet is estimated to be home to more than 80% of life and yet it remains largely unexplored. The rich diversity of marine flora and fauna and its adaptation to the harsh marine environment coupled with new developments in biotechnology, has opened up a new exciting vista for extraction of bioactive products of use in medicine. In this study inhibitory activity of a marine bacterium isolated from gut of ribbonfish was studied against pathogenic and environmental isolates of Vibrio species. This strain was identified as Pseudomonas stutzeri and it was found active against V. harveyi (luminescent bacteria), V. cholerae, V. alginolyticus, V. damseal, V. fluvialis. The antibacterial substance produced by Pseudomonas stutzeri was soluble in organic solvent and closely bound to external surface of bacterial cells. Reduction of the absorbance of the V. cholera cell suspension was observed when log phase cells of V. cholerae were treated with MIC and 4xMIC concentration of crude extract of Pseudomonas stutzeri.

Seasonal changes, life cycle, and production of a psychrophilic chironomid (Propsilocerus akamusi) in a Chinese Lake

Seasonal changes, life cycle, and production of a psychrophilic Chironomidae species, Propsilacerus akamusi (Tokunage), were studied in eutrophic Lake Donghu. The P akamusi population was characterized by a single annual reproduction period during late November to December, and the larval growth mainly occurred in winter. Most of P akamusi were univoltine, while some of them came to emergence in two years or more. The average density and biomass were 318.9 ind./m(2) and 0.57 g dry weight /m(2) during January 1998 to June 2000, respectively, but these values did not include any summer measurement,; since the larvae aestivated in the deep sediment layer and could not be sampled routinely in summer. The annual production of P akamusi was 2.73g dry weight/m(2), and the corresponding production/biomass ratio was 4.60.

Lysis of Aphanizomenon flos-aquae (Cyanobacterium) by a bacterium Bacillus cereus

A phytoplankton-lytic (PL) bacterium, Bacillus cereus, capable of lysing the bloom-forming cyanobacterium. Aphanizomenon flos-aquae was isolated from Lake Dianchi of Yunnan province, China. This bacterium showed lytic activities against a wide range of cyanobacteria/algae, including A. flos-aquae, Microcystis viridis, Microcystis wesenbergi, Microcystis aeruginosa, Chlorella ellipsoidea, Oscillatoria tenuis, Nostoc punctiforme, Anabaena flos-aquae, Spirulina maxima, and Selenastrum capricornutum. Chlorophyll a contents, phycocyanin contents, and photosynthetic activities of the A. flos-aquae decreased evidently in an infected culture for a period. Bacterium B. cereus attacked rapidly A. flos-aquae cells by cell-to-cell contact mechanism. It was shown that the lysis of A. flos-aquae began with the breach of the cyanobacterial cell wall, and the cyanobacterial cell appeared abnormal in the presence of the PL bacterium. Moreover, transmission electron microscope examinations revealed that a close contact between the bacterium and the cyanobacterium was necessary for lysis. Some slime extrusions produced from B. cereus assisted the bacterial cells to be in close association with and lyse the cyanobacterial cells. These findings suggested that this bacterium could play an important role in controlling the Aphanizomenon blooms in freshwaters. (c) 2006 Elsevier Inc. All rights reserved.

The algae-lytic ability of bacterium DC10 and the influence of environmental factors on the ability

A lysing-bacterium DC10, isolated from Dianchi Lake of Yunnan Province, was characterized to be Pseudomonas sp. It was able to lyse some algae well, such as Microcystis viridis, Selenastrum capricomutum, and so on. In this study, it was shown that the bacterium lysed the algae by releasing a substance; the best lytic effects were achieved at low temperatures and in the dark. Different concentrations of CaCl2 and NaNO3 influenced the lytic effects; the ability to lyse algae decreased in the following order: pH 4 > pH 9 > pH 7 > pH 5.5. It was significant to develop a special technology with this kind of bacterium for controlling the bloom-forming planktonic microalgae.

Rhodococcus qingshengii sp nov., a carbendazim-degrading bacterium

A Gram-positive, aerobic, non-motile, mesophilic strain, djl-6(T), able to degrade carbendazim, was isolated from a carbendazim-contaminated soil sample from Jiangsu province, China. The taxonomic position of this isolate was analysed by using a polyphasic approach. Chemotaxonomic analysis including peptidoglycan type, diagnostic sugar composition, fatty acid profile, menaquinones, polar lipids and mycolic acids showed that the characteristics of strain djl-6(T) were in good agreement with those of the genus Rhodococcus. DNA-DNA hybridization showed that it had low genomic relatedness with Rhodococcus baikonurensis DSM 44587(T) (31.8%), Rhodococcus erythropolis DSM 43066(T) (23.8%) and Rhodococcus globerulus DSM 43954(T) (117.7%), the three type strains to which strain djl-6(T) was most closely related based on 16S rRNA gene sequence analysis (99.78, 99.25 and 98.91% similarity, respectively). Based on the phenotypic properties and DNA-DNA hybridization data, strain djl-6(T) (=CGMCC 1.6580(T) =KCTC 19205(T)) is proposed as the type strain of a novel Rhodococcus species, Rhodococcus qingshengii sp. nov.

生防菌BJ1离子辐照突变高效菌株筛选及对黄瓜枯萎病的生防效果；Screening of Mutation High-Yielding Biocontrol Bacterium BJ1 by Ion Beam Irradiation and Effect of Controlling Fusarium oxysporum cucunerinum Disease

枯草芽孢杆菌BJ1是一种在真菌病害防治中发挥重要作用的生防因子,为进一步提高它的抑菌能力,获得生防效果更好的高效菌种,利用不同能量和剂量的12C6+对生防菌BJ1进行了离子辐照处理。研究结果表明:离子辐照生防菌BJ1的最适宜剂量为200~400 Gy,传能线密度(LET)为60 keV/μm;突变菌株的抑菌能力比BJ1提高了2%~21%;不仅防病效果比BJ1提高了17.48%,而且对植物具有更好的促生长作用。

Construction and characterization of a fosmid library for pathogenic bacterium Vibrio anguillarum

Vibrio anguillarum is a common bacterial pathogen in fish. However, little is known about its pathogenic mechanism, in part, because the entire genome has not been completely sequenced. We constructed a fosmid library for V. anguillarum containing 960 clones with an average insert size of 37.7 kb and 8.6-fold genome coverage. We characterized the library by end-sequencing 50 randomly selected clones. This generated 93 sequences with a total length of 57 485 by covering 1.4% of the whole genome. Of these sequences, 58 (62.4%) were homologous to known genes, 30 (32.3%) were genes with hypothetical functions, and the remaining 5 (5.3%) were unknown genes. We demonstrated the utility of this library by PCR screening of 10 genes. This resulted in an average of 6.2 fosmid clones per screening. This fosmid library offers a new tool for gene screening and cloning of V. anguillarum, and for comparative genomic studies among Vibrio species.

Hymeniacidon perleve associated bioactive bacterium Pseudomonas sp NJ6-3-1

Among marine bacteria isolated from the cytotoxic sponge Hymeniacidon perleve, one strain NJ6-3-1 classified as Pseudomonas sp. showed both cytotoxic and antimicrobial activities. Fatty acid analysis indicated that the bacterial strain consists mainly of C16:1, C16:0, C18:1, C18:0, C15:0, C14:0. One unusual 9,10-cyclopropane-C17:0 fatty acid and C26:0 also constitute major components, as well as the existence of squalene, the precursor of triterpenoids. The major metabolites in the culture broth were identified as alkaloids, including diketopiperazines and indole compounds, namely 3,6-diisopropylpiperazine-2,5-dione, 3-benzyl-3-isopropylpiperazine-2,5-dione, 3,6-bis-(2-methylpropyl)-piperazine-2,5-dione, indole-3-carboxaldehyde, indole-3-carboxylic acid methyl ester, indole-3-ethanol, and quinazoline-2,4-dione.

Programmed cell death in Laminaria japonica (Phaeophyta) tissues infected with alginic acid decomposing bacterium

TdT-mediated dUTP-biotin nick end labeling (TUNEL) is a sensitive and valid method for detecting DNA cleavage in programmed cell death (PCD). Using this method, DNA cleavage was observed in Laminaria japonica sporophytic tissues, which were infected with alginic acid decomposing bacterium. It was found that DNA cleavage occurred 5 min after the infection, the fragments with 3'-OH groups of cleaved nuclear DNA increased with time of infection and spread from the infection site. Although no typical DNA ladder (200 bp/ 180 bp) was detected by routine agarose gel electrophoresis, the cleavage of nuclear DNA fragments of 97 similar to 48.5 kb could be detected by pulsed field gel electrophoresis (PFGE). By using CaspGLOW(TM) fluorescein active caspase-3 staining method, caspase-3 activity has been detected in response to the infection of alginic acid decomposing bacterium. Our results are similar to the observations in hypersensitive response (HR) of higher plant, suggesting that the rapid cell death of L. japonica infected by alginic acid decomposing bacterium might be involved in PCD, and indicating that the occurrence of PCD is an active defense process against the pathogen's infection.