Beneficial properties of lactic acid bacteria isolated from a Rana catesbeiana hatchery

This work addresses the selection of potentially probiotic lactic acid bacteria (LAB) to be used in raniculture. Thus, strains belonging to the genera Pediococcus pentosaceus, Leuconostoc mesenteroides, Lactococcus lactis and Enterococcus faecium isolated from a Rana catesbeiana hatchery were evaluated for their inhibitory properties against RLS-associated pathogens ( Proteus vulgaris, Pseudomonas aeruginosa, Staphylococcus epidermidis ) and food-borne bacteria. Cell-free supernatants of LAB strains inhibited the growth of at least one of the pathogens by organic acids, but L. lactis CRL 1584 also produced a bacteriocin-like metabolite. The ability of LAB strains to produce H₂O₂ in MRS+TMB medium was also studied. Seventy-eight to ninety six per cent of the strains showed some level of H₂O₂ production. Moreover, different organic solvents were used to determine the hydrophobicity and Lewis acid/base characteristic of LAB strain surfaces. Most of the strains presented hydrophilic properties, but no acidic or basic surface characters. However, some strains isolated from the skin showed a high degree of hydrophobicity and basic components in the cell surface due to their adhesion to chloroform. These properties were not observed in LAB from balanced feed and freshwater. Taking into account general guidelines and the beneficial properties studied, five strains were selected as potential candidates to be included in a probiotic for raniculture.     

Effect of starvation on the performance of baby octopus (Robsonella fontaniana) paralarvae

The aim of this study was to evaluate the effect of short‐ and long‐term starvation on paralarvae from hatching and compare to fed paralarvae. In the continuous starvation treatments, paralarvae at hatching were left without food as independent treatments for 5, 8, 9, 10, 11, 12, 13 and 15 days. In the fed treatments, the newly hatched paralarvae were fed for five and eight days; then each group was left in starvation as independent treatments for 3, 5 and 7 days. After any experimental starvation period, the paralarvae were fed for five more days to evaluate their recovery. Paralarvae exposed to continuous starvation from hatching endured up to 8 days after hatching (8 SDAH) showing significant recovery. Its survival decreased proportionally to the days of starvation, without any recovery after 12 DAH. Fed paralarvae (5 and 8 FDAH), resulted in significant differences accordingly to the length of the starvation period. Paralarvae left under permanent starvation showed a noticeable decrease in their arm/mantle length ratio and an atrophy of the digestive gland was observed. Amino acids were used primarily as energy source by paralarvae, particularly in the second week of starvation. It is discussed the critical age to avoid deleterious effects of starvation on paralarvae.     

Rapid sequence modification in the highly polymorphic region (HPR) of the hemagglutinin gene of the infectious salmon anaemia virus (ISAV) suggests intra-segmental template switching recombination

The ISAV has a genome composed of eight segments of (–)ssRNA, segment 6 codes for the hemagglutinin–esterase protein, and has the most variable region of the genome, the highly polymorphic region (HPR), which is unique among orthomyxoviruses. The HPR has been associated with virulence, infectivity and pathogenicity. The full length of the HPR is called HPR0 and the strain with this HPR is avirulent, in contrast to strains with deleted HPR that are virulent to varying degrees. The molecular mechanism that gives rise to the different HPRs remains unclear. Here, we studied in vitro the evolution of reassortant recombinant ISAV (rISAV) in Atlantic salmon head kidney (ASK) cells. To this end, we rescued and cultivated a set of rISAV with different segment 6-HPR genotypes using a reverse genetics system and then sequencing HPR regions of the viruses. Our results show rapid multiple recombination events in ISAV, with sequence insertions and deletions in the HPR, indicating a dynamic process. Inserted sequences can be found in four segments of the ISAV genome (segments 1, 5, 6, and 8). The results suggest intra-segmental heterologous recombination, probably by class I and class II template switching, similar to the proposed segment 5 recombination mechanism.     

Assessment of dietary lipid sources in tropical gar,Atractosteus tropicus larvae: Growth parameters and intermediary lipogenic gene expression

Five experimental diets containing different lipid sources, fish oil (D1), soybean lecithin (D2), corn oil (D3), canola oil (D4) and olive oil (D5), were evaluated in Atractosteus tropicus larvae for the relative gene expression of the enzymes fatty acid synthase (fas), acetyl‐CoA carboxylase 1 (acc1) and carnitine palmitoyltransferase 1C (cpt1c), in addition to their effects on larval growth, survival and cannibalism during a 30‐day feeding trial. Higher growth and survival were obtained in treatments D1 and D2, and lower performance in diets D3, D4 and D5. The highest levels of expression of fas and acc1 occurred in larvae fed with D1, which contained high amounts of n‐3 long‐chain polyunsaturated fatty acids (LC‐PUFA), mainly DHA and EPA FA are regulators of lipogenesis. The higher cpt1c expression in plant‐based diets is attributed to the fact that these diets are rich in α‐linolenic acid (ALA) and low DHA, EPA and ARA levels that favour ß‐oxidation. In conclusion, the diets with fish oil (D1) and soybean lecithin (D2) were the best treatments for larval growth, survival and cannibalism and thus appear to meet both lipid and energy requirements of A. tropicus larvae, meanwhile the use of vegetable oils influences the expression of intermediary lipogenic genes.     

Evidence for the facultative intracellular behaviour of the fish pathogen Vibrio ordalii

Vibrio ordalii is an extracellular, Gram‐negative bacterium that produces vibriosis in salmonids. While pathogenesis is not fully understood, this bacterium has numerous likely genes for adhesion, colonization, invasion factors and, as recently suggested, intracellular behaviour. Therefore, this study aimed to clarify possible intracellular behaviour for V. ordalii Vo‐LM‐18 and ATCC 33509^T in the fish‐cell lines SHK‐1 and CHSE‐214. Confocal microscopy revealed Vo‐LM‐18 and ATCC 33509^T inside cytoplasm in both fish‐cell lines at 4 hr post‐inoculation (hpi). At 8 and 16 hpi, the proportion of fish cells invaded by both strains increased. Moreover, intracellular V. ordalii were observed after 8 hpi inside mouse embryonic fibroblasts (MEF), demonstrating that entry was not due to a cellular phagocytosis process. Flow cytometry confirmed immunocytochemistry results, with both V. ordalii evidencing statistically significant differences in the number of infected cells between 8 and 16 hpi. Interestingly, V. ordalii infection did not significantly damage fish cells, as determined by LDH liberation. Viable counts at 8 hpi detected, on average for both lines, 176 ± 47 CFU/ml of culturable intracellular Vo‐LM‐18 and ATCC 33509^T cells. These in vitro findings support the facultative intracellular behaviour of V. ordalii and may be of importance for understanding pathogenicity and survival in aquatic environments.     

Putting the pieces together: Recent growth,nutritional condition,and mortality of Engraulis anchoita larvae in the Southwest Atlantic

Dynamics of clupeiform fish populations such as anchovy are frequently impacted by environmental variations which can affect the success of the species recruitment. Herein, we have analyzed recent otolith growth rate, RNA/DNA nutritional condition index (sRD), and mortality rate of argentine anchovy larvae Engraulis anchoita from three different nursery areas in the Southwest Atlantic. We have evaluated the relationship between the environmental variables (abundance of copepod nauplii, temperature, chlorophyll‐a concentration, and abundance of E. anchoita larvae) and larval endogenous variables (size, weight, age, and otolith radius) to sRD and recent growth rate. Fast larval growth rates were observed toward the northern sector of the studied area, characterized by higher temperature. High values of sRD were associated with higher nauplii abundance in the proximity of coastal fronts. The larvae with the lowest growths and lowest minimum values of nutritional condition coincided with the area where there was less abundance of nauplii and higher larval mortality. Larval size and nauplii abundance were positive explanatory factors for both recent growth rate and sRD index. Temperature had a positive effect on recent growth rate and a negative effect on sRD index. This condition index was poorly explained in terms of model fit in comparison with the growth model. The results herein provided could be significant to better understand the recruitment of the species, as to determining favorable areas for the growth and survival of anchovy larvae.     

Ecosystem‐based management of Amazon fisheries and wetlands

Infrastructure development and overfishing in the Amazon make it imperative to define adequate scales for the ecosystem‐based management of commercial fisheries and the wetlands on which they depend. We mapped fisheries and fish ecology data from Brazil, Peru, Bolivia and Colombia to an explicit GIS framework of river basins and mainstems. Migratory species account for more than 80% of the known maximum catches of commercial fisheries across the Amazon. Of these migratory species, we nominated six long‐distance migratory fish taxa as flagship species to define the two main commercial fishery regions. The migrations of at least one goliath catfish species define a large‐scale longitudinal link joining the Andes, Amazon Lowlands and Amazon River estuary. Migratory Characiforms demonstrate interbasin wetland connectivity between nutrient‐rich and nutrient‐poor rivers over at least 2 million km², or about one‐third of the Amazon Basin. We show that flooded forest area is the most important wetland variable explaining regional variations in migratory characiforme biomass as indicated by maximum annual fishery catches. The sustainable management of Amazon fisheries will require transnational cooperation and a paradigm shift from local community management alone to a more integrated approach that considers both rural and urban consumers and challenges, and the realistic life histories of migratory species.     

Effects of temperature and body weight on growth rate and feed conversion ratio in turbot (Scophthalmus maximus)

Six size groups of hatchery produced turbot (2–499 g) were reared at six constant temperatures (10–25 °C). The results were used to develop a mathematical model for growth rate and feed conversion in turbot. At each temperature there were linear relationships between logarithms of (a) specific growth rate (G%/day) and body weight (W g), and (b) feed conversion ratio (FCR) and W. The slopes of the regressions were linearly related to temperature and the intercepts of the regression changed with temperature according to a second order polynomial. The optimal temperature for growth (T_opt.G) is predicted to decline with increasing body weight: T_opt.G = 22.45 − 0.713lnW, i.e. 22.5, 20.8, 19.1 and 17.5 °C for 1, 10, 100 and 1000 g fish, respectively, and the growth rates of these fish sizes at their optimal temperature (G_max) were predicted to be 7.63, 2.73, 1.03 and 0.40%/day, respectively. The optimal temperature for feed conversion (T_opt.FCR) is also predicted to decline with increasing fish size: T_opt.FCR = 18.80 − 0.625lnW, i.e. 18.8, 17.4, 15.9 and 14.5 °C for 1, 10, 100 and 1000 g fish, respectively, and the predicted feed conversions of these fish sizes (FCR_min) were 0.44, 0.56, 0.68 and 0.82 for the present feed types. The growth model predicts that 2 g turbot can reach 743 and 619 g in 1 year when reared at T_opt.G and T_opt.FCR, respectively.     

Identification of genetic sources with attenuated <Emphasis Type="Italic">Tomato chlorosis virus</Emphasis>-induced symptoms in <Emphasis Type="Italic">Solanum</Emphasis> (section <Emphasis Type="Italic">Lycopersicon</Emphasis>) germplasm

The whitefly-transmitted Tomato chlorosis virus (ToCV) (genus Crinivirus) is associated with yield and quality losses in field and greenhouse-grown tomatoes (Solanum lycopersicum) in South America. Therefore, the search for sources of ToCV resistance/tolerance is a major breeding priority for this region. A germplasm of 33 Solanum (Lycopersicon) accessions (comprising cultivated and wild species) was evaluated for ToCV reaction in multi-year assays conducted under natural and experimental whitefly vector exposure in Uruguay and Brazil. Reaction to ToCV was assessed employing a symptom severity scale and systemic virus infection was evaluated via RT-PCR and/or molecular hybridization assays. A subgroup of accessions was also evaluated for whitefly reaction in two free-choice bioassays carried out in Uruguay (with Trialeurodes vaporariorum) and Brazil (with Bemisia tabaci Middle-East-Asia-Minor1—MEAM1?=?biotype B). The most stable sources of ToCV tolerance were identified in Solanum habrochaites PI 127827 (mild symptoms and low viral titers) and S. lycopersicum ‘LT05’ (mild symptoms but with high viral titers). These two accessions were efficiently colonized by both whitefly species, thus excluding the potential involvement of vector-resistance mechanisms. Other promising breeding sources were Solanum peruvianum (sensu lato) ‘CGO 6711’ (mild symptoms and low virus titers), Solanum chilense LA1967 (mild symptoms, but with high levels of B. tabaci MEAM1 oviposition) and Solanum pennellii LA0716 (intermediate symptoms and low level of B. tabaci MEAM1 oviposition). Additional studies are necessary to elucidate the genetic basis of the tolerance/resistance identified in this set of Solanum (Lycopersicon) accessions.