An Investigation into OccasionalWhite Spot Syndrome Virus Outbreak in Traditional Paddy CumPrawn Fields in India

A yearlong (September 2009–August 2010) study was undertaken to find out possible reasons for occasional occurrence of White Spot Syndrome Virus (WSSV) outbreak in the traditional prawn farms adjoining Cochin backwaters. Physicochemical and bacteriological parameters of water and sediment from feeder canal and four shrimp farms were monitored on a fortnightly basis. The physicochemical parameters showed variation during the two production cycles and between the farms studied. Dissolved oxygen (DO) content of water fromfeeder canal showed low oxygen levels (as low as 0.8mg/L) throughout the study period. There was no disease outbreak in the perennial ponds. Poor water exchange coupled with nutrient loading from adjacent houses resulted in phytoplankton bloom in shallow seasonal ponds which led to hypoxic conditions in early morning and supersaturation of DO in the afternoon besides considerably high alkaline pH. Ammonia levels were found to be very high in these ponds.WSSV outbreak was encountered twice during the study leading to mass mortalities in the seasonal ponds. The hypoxia and high ammonia content in water and abrupt fluctuations in temperature, salinity and pH might lead to considerable stress in the shrimps triggeringWSSV infection in these traditional ponds

Prevalence of Aeromonas hydrophila in fish and prawns from the seafood market of Coimbatore, South India

Five hundred and thirty six samples offishes and 278 prawn samples from the major fish market ofCoimbatore, South India, were analysed for the prevalence of Aeromonas hydrophila over a period of2 years (June 1997–May 1999). The prevalence level of A. hydrophila varied from 17.62% in prawns to 33.58% in fishes. More than 30% of the popular table fishes such as Sardinella longiceps, Rastrelliger kanagurta, Mugil cephalus and Caranx sexfasciatus were tested positive for this organism. Among the different species of the prawns analysed, Penaeus semisulcatus showed higher incidence (23.52%). Seasonal variation in the prevalence levels of A. hydrophila in fish and prawns revealed a higher prevalence during the monsoon season during 1997–98 and 1998–99. Of the different body parts of the fishes analysed for A. hydrophila, the intestinal samples showed higher prevalence (38.43%), followed by body surface (32.46%) and gill (29.10%). Considering the psychrotrophic nature and role of A. hydrophila as a pathogen ofemerging importance, the considerably high levels ofthis organism in a popular food item such as fish and prawn raises serious concern

A brackishwater isolate of Pseudomonas PS-102, a potential antagonistic bacterium against pathogenic vibrios in penaeid and non-penaeid rearing systems

A Pseudomonas sp PS-102 recovered from Muttukkadu brackish water lagoon, situated south of Chennai, showed significant activity against a number of shrimp pathogenic vibrios. Out of the 112 isolates of bacterial pathogens comprising Vibrio harveyi, V. vulnificus, V. parahaemolyticus, V. alginolyticus, V. fluvialis, and Aeromonas spp, 73% were inhibited in vitro by the cell-free culture supernatant of Pseudomonas sp PS-102 isolate. The organism produced yellowish fluorescent pigment on King's B medium, hydrolysed starch and protein, and produced 36.4% siderophore units by CAS assay and 32 μM of catechol siderophores as estimated by Arnow's assay. The PS-102 isolate showed wide ranging environmental tolerance with, temperatures from 25 to 40 °C, pH from 6 to 8, salinity from 0 to 36 ppt, while the antagonistic activity peaked in cultures grown at 30 °C, pH 8.0 and at 5 ppt saline conditions. The antagonistic activity of the culture supernatant was evident even at 30% v / v dilution against V. harveyi. The preliminary studies on the nature of the antibacterial action indicated that the antagonistic principle as heat stable and resistant to proteolytic, lipolytic and amylolytic enzymes. Pseudomonas sp PS 102 was found to be safe to shrimp when PL-9 stage were challenged at 107 CFU ml−1 and by intramuscular injection into of ∼5 g sub-adults shrimp at 105 to 108 CFU. Further, its safety in a mammalian system, tested by its pathogenicity to mice, was also determined and its LD50 to BALB/c mice was found to be 109 CFU. The results of this study indicated that the organism Pseudomonas sp PS 102 could be employed as a potential probiont in shrimp and prawn aquaculture systems for management and control of bacterial infections

Development of a cell culture system from the ovarian tissue of African catfish Clarias gariepinus

A growth medium with Leibovitz-15 L-15.as the base, supplemented with foetal bovine serum 10% vrv., fish muscle extract 10% vrv., prawn muscle extract 10% vrv., lectin concanavalin A. 0.02 mg mly1., lipopolysaccharide 0.02 mg mly1., glucose D 0.2 mg mly1., ovary extract 0.5% vrv.and prawn haemolymph 0.5%. has been formulated with 354"10 mOsm for the development and maintenance of a cell culture system from the ovarian tissue of African catfish, Clarias gariepinus. For its subculturing, a cell dissociationrextracting solution, composed of equal portions of trypsin phosphate versene glucose TPVG. containing 0.0125% wrv.trypsin and 25% vrv.non-enzymatic cell dissociation solution 1 and 2, has also been developed with which the cell culture can be passaged 15 times after which they cease to multiply and consequently perish. The cell cultures can be maintained for 12–15 days without fluid change between the passages. This is the first report of a cell culture system from the ovarian tissues of African catfish

Development of nitrifying bacterial consortia for immobilizing in nitrifying bioreactors designed for penaeid and non-penaeid larval rearing systems in the tropics

Two ammonia oxidizing (AMOPCU-1 and AMONPCU-1) and two nitrite oxidizing (NIOPCU-1 and NIONPCU-1) consortia for activating nitrifying bioreactors and thereby establishing nitrification in penaeid and non-penaeid hatchery systems were developed by enrichment. For further amplification of the consortia a simple medium having seawater (either salinity 30 ‰ or 15 ‰) as base, supplemented with NH4+-N/NO2--N and PO4- and pH adjusted to 8 was identified. During the amplification in a fermentor the consortia exhibited excessive wall growth and diminished their yield coefficient posing difficulty in harvesting the cells completely. The consortia consisted of both Gram negative and Gram-positive bacterial cells embedded in a mucilaginous matrix of glycocalyx - like material presumably composed of polysaccharides. The consortia besides being useful in activating nitrifying bioreactors developed for shrimp/prawn hatchery systems can also be used as bioaugmentors in the bioremediation of ammonia and nitrite toxicity in aquaculture systems.

Fish flesh agar medium - a suitable experimental medium for the detection of spoilage bacteria

The spoilage characteristics of bacterial strains were studied by growing them at 28 _+ 2 °C in agar and broth media prepared with sterile fish and prawn flesh homogenates. The percentage of spoilers found among the bacterial isolates tested, as shown by odour production and halo zone formation, was independent of the source of flesh used. Indole and fluorescent pigment production were also observed in the broth. Pseudomonas, Vibrio and Acinetobacter exhibited faster growth in flesh media than in the usual artificial media. Decrease of protein and lipid concentration in the clear zone of agar media suggests the utilization of the available substrate by spoilage bacteria.

Characterization and Pathogenicity of Vibrio cholerae and Vibrio vulnificus from Marine environments

The genus Vibrioof the family Vibrionaceae are Gram negative, oxidasepositive, rod- or curved- rodshaped facultative anaerobes, widespread in marine and estuarine environments. Vibrio species are opportunistic human pathogens responsible for diarrhoeal disease, gastroenteritis, septicaemia and wound infections and are also pathogens of aquatic organisms, causing infections to crustaceans, bivalves and fishes. In the present study, marine environmental samples like seafood and water and sediment samples from aquafarms and mangroves were screened for the presence of Vibrio species. Of the134 isolates obtained from the various samples, 45 were segregated to the genus Vibrio on the basis of phenotypic characterization.like Gram staining, oxidase test, MoF test and salinity tolerance. Partial 16S rDNA sequence analysis was utilized for species level identification of the isolates and the strains were identified as V. cholerae(N=21), V. vulnificus(N=18), V. parahaemolyticus(N=3), V. alginolyticus (N=2) and V. azureus (N=1). The genetic relatedness and variations among the 45 Vibrio isolates were elucidated based on 16S rDNA sequences. Phenotypic characterization of the isolates was based on their response to 12 biochemical tests namely Voges-Proskauers’s (VP test), arginine dihydrolase , tolerance to 3% NaCl test, ONPG test that detects β-galactosidase activity, and tests for utilization of citrate, ornithine, mannitol, arabinose, sucrose, glucose, salicin and cellobiose. The isolates exhibited diverse biochemical patterns, some specific for the species and others indicative of their environmental source.Antibiogram for the isolates was determined subsequent to testing their susceptibility to 12 antibiotics by the disc diffusion method. Varying degrees of resistance to gentamycin (2.22%), ampicillin(62.22%), nalidixic acid (4.44%), vancomycin (86.66), cefixime (17.77%), rifampicin (20%), tetracycline (42.22%) and chloramphenicol (2.22%) was exhibited. All the isolates were susceptible to streptomycin, co-trimoxazole, trimethoprim and azithromycin. Isolates from all the three marine environments exhibited multiple antibiotic resistance, with high MAR index value. The molecular typing methods such as ERIC PCR and BOX PCR revealed intraspecies relatedness and genetic heterogeneity within the environmental isolatesof V. cholerae and V. vulnificus. The 21 strains of V. choleraewere serogroupedas non O1/ non O139 by screening for the presence O1rfb and O139 rfb marker genes by PCR. The virulence/virulence associated genes namely ctxA, ctxB, ace, VPI, hlyA, ompU, rtxA, toxR, zot, nagst, tcpA, nin and nanwere screened in V. cholerae and V. vulnificusstrains.The V. vulnificusstrains were also screened for three species specific genes viz., cps, vvhand viu. In V. cholerae strains, the virulence associated genes like VPI, hlyA, rtxA, ompU and toxR were confirmed by PCR. All the isolates, except for strain BTOS6, harbored at least one or a combination of the tested genes and V. choleraestrain BTPR5 isolated from prawn hosted the highest number of virulence associated genes. Among the V. vulnificusstrains, only 3 virulence genes, VPI, toxR and cps, were confirmed out of the 16 tested and only 7 of the isolates had these genes in one or more combinations. Strain BTPS6 from aquafarm and strain BTVE4 from mangrove samples yielded positive amplification for the three genes. The toxRgene from 9 strains of V. choleraeand 3 strains of V. vulnificus were cloned and sequenced for phylogenetic analysis based on nucleotide and the amino acid sequences. Multiple sequence alignment of the nucleotide sequences and amino acid sequences of the environmental strains of V. choleraerevealed that the toxRgene in the environmental strains are 100% homologous to themselves and to the V. choleraetoxR gene sequence available in the Genbank database. The 3 strains of V. vulnificus displayed high nucleotide and amino acid sequence similarity among themselves and to the sequences of V. cholerae and V. harveyi obtained from the GenBank database, but exhibited only 72% homology to the sequences of its close relative V. vulnificus. Structure prediction of the ToxR protein of Vibrio cholerae strain BTMA5 was by PHYRE2 software. The deduced amino acid sequence showed maximum resemblance with the structure of DNA-binding domain of response regulator2 from Escherichia coli k-12 Template based homology modelling in PHYRE2 successfully modelled the predicted protein and its secondary structure based on protein data bank (PDB) template c3zq7A. The pathogenicity studies were performed using the nematode Caenorhabditiselegansas a model system. The assessment of pathogenicity of environmental strain of V. choleraewas conducted with E. coli strain OP50 as the food source in control plates, environmental V. cholerae strain BTOS6, negative for all tested virulence genes, to check for the suitability of Vibrio sp. as a food source for the nematode;V. cholerae Co 366 ElTor, a clinical pathogenic strain and V. cholerae strain BTPR5 from seafood (Prawn) and positive for the tested virulence genes like VPI, hlyA, ompU,rtxA and toxR. It was found that V. cholerae strain BTOS6 could serve as a food source in place of E. coli strain OP50 but behavioral aberrations like sluggish movement and lawn avoidance and morphological abnormalities like pharyngeal and intestinal distensions and bagging were exhibited by the worms fed on V. cholerae Co 366 ElTor strain and environmental BTPR5 indicating their pathogenicity to the nematode. Assessment of pathogenicity of the environmental strains of V. vulnificus was performed with V. vulnificus strain BTPS6 which tested positive for 3 virulence genes, namely, cps, toxRand VPI, and V. vulnificus strain BTMM7 that did not possess any of the tested virulence genes. A reduction was observed in the life span of worms fed on environmental strain of V. vulnificusBTMM7 rather than on the ordinary laboratory food source, E. coli OP50. Behavioral abnormalities like sluggish movement, lawn avoidance and bagging were also observed in the worms fed with strain BTPS6, but the pharynx and the intestine were intact. The presence of multi drug resistant environmental Vibrio strainsthat constitute a major reservoir of diverse virulence genes are to be dealt with caution as they play a decisive role in pathogenicity and horizontal gene transfer in the marine environments.

Kinetic Analysis of Gill (Na+,K+)-ATPase Activity in Selected Ontogenetic Stages of the Amazon River Shrimp, (Decapoda, Palaemonidae): Interactions at ATP- and Cation-Binding Sites

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Size-related shifts in dietary composition of Centropomus parallelus (Perciformes: Centropomidae) in an estuarine ecosystem of the southeastern coast of Brazil

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Intra- and extracellular osmotic regulation in the hololimnetic Caridea and Anomura: a phylogenetic perspective on the conquest of fresh water by the decapod Crustacea

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Tratamento de efluentes de carcinicultura por macrófitas aquáticas flutuantes

The efficiency of systems composed of two species of floating aquatic macrophyte (Eichhornia crassipes and Pistia stratiotes) to treat effluents from shrimp culture was evaluated in this research. The effluent originated from a pond populated with Macrobrachium amazonicum. Treatment systems consisted of 12 experimental tanks with aquatic macrophyte and three tanks without plants (control). Water samples were collected from the fresh water supply and before and after passing through the treatment systems. There are no differences in the removal of nutrients between the two species. The higher nutrient removal was observed for total phosphorus (41.9% by control; 71.6% by E. crassipes; 69.9 by P. stratiotes; 72.5% by E. crassipes + P. stratiotes and 72.1 by P. stratiotes + E. crassipes) and turbidity (30.6% by control; 80.2% by E. crassipes; 75.2 by P. stratiotes; 79.8% by E. crassipes + P. stratiotes and 81.5 by P. stratiotes + E. crassipes). The systems containing aquatic macrophytes were efficient in nitrogen and phosphorus removal from M. amazonicum culture. © 2008 Sociedade Brasileira de Zootecnia.

Reproductive cycle and fecundity of Potimirim glabra (Kingsley, 1954) (Caridea, Atyidae) from a littoral stream

The reproductive cycle of Potimirim glabra, especially the female size at sexual maturity, reproductive period and fecundity, was studied at two sites of one of the northern littoral streams in the state of São Paulo, Brazil. Prawns were collected monthly for one year. The minmum size at sexual maturity was estimated from the smallest ovigerous female (cephalothorax length = CL), and the reproductive period was determined monthly by examining the relative frequency of ovigerous females. Size comparisons between sampling sites was carried out using the Mann-Whitney u test. The relationship between environmental factors and reproductive aspects was verified using the Spearman's rank correlation. At Site 1, the collected ovigerous females had a CL mean size of 4.24±0.36 mm (n=481); at Site 2, ovigerous females (n=391) had a CL mean size of 5.48±0.49 mm. The reproductive pattern in Camburi is seasonal, with the highest frequency of ovigerous females in February. The frequency of ovigerous females was positively correlated to organic matter content in the substratum, rainfall and temperature. Increased percentage organic matter may indicate greater food availability whereas higher rainfall increases the velocity of the current, thereby aiding the larvae, which depend on brackish water to complete their development, to reach the estuarine region. The temperature acts on the speed of larval development. At Site 1, the mean fecundity obtained was 202±72.31 eggs (90 females; CL from 2.7 to 4.7mm PL), whereas at Site 2, it was 433.3±120.7 eggs (46 females; CL from 4.0 to 5.5mm PL). © 2010 Balaban.

Influência de uma piscicultura em tanquesrede na dieta e atributos biológicos de Galeocharax knerii na represa de Chavantes, Brasil

The aim of the present study was to evaluate the diet and biological attributes of the population of Galeocharax knerii residing near net cage fish farming activities in the Chavantes reservoir (Paranapanema River, Brazil) to check their possible impacts. Samples were collected from two populations: one near the net cages (NC) and one from an area not influenced by these cages denominated the reference site (RS). Monthly sampling was carried out from March 2008 to February 2009. Fish were caught with a standardized effort using gill nets deployed for 14 hours. The alimentary index (AI) and degree of repletion (RD) were calculated to determine diet composition. Analyses of the sex ratio and the gonadosomatic index (GSI) were also performed. The calculations of AI revealed that fish wastes constituted the most frequent food item in the diet in both study areas (NC = 70.43; RS = 87.55), followed by the consumption of Apareiodon affinis (AI = 29.56), which was abundant near the NC, and prawn at the reference site (AI = 12.28). The sex ratio differed from 1:1 and mature individuals were only found in the population near the NC. The findings demonstrate that G. knerii indirectly benefits from the input of organic matter, using small fish as its main food resource. We conclude that the activities of fish farming influence diet and biological attributes of the species G. knerii, evidenced mainly by higher feeding activity, numerical abundance and biomass in the area of the cages.

Rastreabilidade de camarão amazônico proveniente de cultivo e ambiente natural utilizando a técnica de isótopos estáveis

Pós-graduação em Aquicultura - FCAV

Sistemas de criação para o Acará-bandeira Pterophyllum scalare

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)