Existence of two O-serotypes in the fish pathogen Pseudomonas anguilliseptica

The serological characteristics of a group of 32 Pseudomonas anguilliseptica strains isolated in Spain from seabream (Sparus aurata) and turbot (Scophthalmus maximus) were compared with a total of 18 collection strains of this bacterial species with different geographical and host origin. The employment of different techniques, including slide agglutination, microagglutination and dot blot, allowed us to establish two serological groups, one comprising practically all the eel isolates, and the other including the majority of isolates from other fish species. The study of the lipopolysaccharides (LPS) and outer membrane proteins (OMP) corroborated these results, indicating that the serological differences among strains are due to the somatic antigen and not to antigenic determinants of protein nature. Therefore, a serological scheme of two "O" serotypes is proposed for P. anguilliseptica. The results obtained will be of importance for epidemiological studies as well as for the design of adequate vaccine formulations.     

Characterization of Edwardsiella tarda strains isolated from turbot, Psetta maxima (L.)

The biochemical, serological and molecular characteristics of a group of 21 Edwardsiella tarda strains isolated from turbot, Psetta maxima, in two different areas of Europe were analysed and compared with a total of 13 strains of this bacterial species with different geographical and host origins. All the turbot isolates were biochemically identical to the E. tarda strains included as reference. The use of different techniques including microagglutination, dot blot and Western blot of lipopolysaccharides allowed us to determine that all the turbot isolates constitute an homogeneous and distinctive serological group. Genetic analysis by randomly amplified polymorphic DNA (RAPD) analysis demonstrated that although the E. tarda strains from turbot were compiled in a unique group using the primers P3 and P6, two clonal lineages could be detected when oligonucleotides P4 and P5 were employed.     

Effectiveness of a divalent vaccine for sole, Solea senegalensis (Kaup), against Vibrio harveyi and Photobacterium damselae subsp. piscicida

The protection of cultured sole, Solea senegalensis, against Vibrio harveyi and Photobacterium damselae subsp. piscicida was evaluated following the use of a divalent vaccine prepared with formalized whole cells and extracellular products of virulent strains of both pathogenic microorganisms and administered by the immersion route. Two prolonged immersions of 5-10 g fish in the divalent bacterin at a 1-month interval gave high levels of protection similar to those obtained when the respective monovalent vaccines were administered by the intraperitoneal route [relative percentage of survival (RPS) values >70%], which indicates that the former procedure can be a useful strategy with small fish. The high protection afforded by the divalent vaccine in sole lasted for 4 months after which the RPS values against both pathogens decreased significantly.     

Genetics of resistance to kernel infection by Aspergillus flavus in maize *

Although kernel infection by Aspergillus flavus Link ex Fries and subsequent pre-harvest aflatoxin contamination of maize (Zea mays L.) grain are major production problems in the south-eastern United States and elsewhere in the world, limited progress has been made in developing and identifying sources for resistance. Genetics of kernel infection by A. flavus remains poorly understood. A 10-parent diallel experiment was conducted in 1992 and 1994 to study the genetic nature of percentage kernel infection (PKI) by A. flavus. General combining ability (GCA), specific combining ability (SCA), and reciprocal mean squares for PKI were significant. The GCA and SCA sums of squares were about equal. The GCA, SCA, and reciprocal effects varied across years, which implied that percentage kernel infection was greatly influenced by environments. The percentage kernel infection was always greater when Mo 17 was the male parent in a cross than when it was the female parent. The percentage kernel infection mean for female Mo 17 was 38.5% lower than that for male Mol7. Similarly, percentage kernel infection mean for female L668 was 23.7% lower than that for male L668. The use of inbred lines L729 and B73 as female parents should be avoided, as they showed significant, positive maternal reciprocal effects.     

Genotype selection via a new yield-stability statistic in maize yield trials

Summary Genotype × environment (GE) interaction complicates selection of superior genotypes across environments. The main objective of this study was to select maize (Zea mays L.) genotypes via a new yield-stability (YS_i) statistic in yield trials conducted in Albania. Another objective was to estimate contribution of environmental index (% MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaaiaacaqabeaadaqaaqaaaOqaaiqadIfagaqeaa% aa!3851!\[\bar X\]_·j – % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaaiaacaqabeaadaqaaqaaaOqaaiqadIfagaqeaa% aa!3851!\[\bar X\]_.., where % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaaiaacaqabeaadaqaaqaaaOqaaiqadIfagaqeaa% aa!3851!\[\bar X\]_·j is mean of all genotypes in the jth environment and % MathType!MTEF!2!1!+-% feaafiart1ev1aaatCvAUfeBSjuyZL2yd9gzLbvyNv2CaerbuLwBLn% hiov2DGi1BTfMBaeXatLxBI9gBaerbd9wDYLwzYbItLDharqqtubsr% 4rNCHbGeaGak0dh9WrFfpC0xh9vqqj-hEeeu0xXdbba9frFj0-OqFf% ea0dXdd9vqaq-JfrVkFHe9pgea0dXdar-Jb9hs0dXdbPYxe9vr0-vr% 0-vqpWqaaeaabaGaaiaacaqabeaadaqaaqaaaOqaaiqadIfagaqeaa% aa!3851!\[\bar X\]is mean of all genotypes across all environments), minimum temperature, maximum temperature, preseason rainfall, rainfall during the growing season, and relative humidity to GE interaction by determining heterogeneity (nonadditivity) attributable to each of these environmental factors. In five of eight trials, heterogeneity due to environmental index was significant. Heterogeneity due to the other environmental factors was not significant in any trial. A comparison of _i ² (stability-variance statistic derived from total GE interaction) and s _i ² (stability-variance statistic derived from residual GE interaction following removal of heterogeneity due to encovariate) helped identify genotypes that performed stably or unstably because of a linear effect of environmental index. In three of the five trials showing significant heterogeneity due to environmental index, the YS_i statistic selected a reduced number of unstable genotypes as compared with selection based solely on yield. However, the circumstances or conditions under which YS_i and solely yield-based method select the same or different genotypes are not fully understood.     

Streptococcus phocae, an emerging pathogen for salmonid culture

This work describes the characterization of the causal agent of disease outbreaks that, from 1999, occurred repeatedly during the summer months (temperatures higher than 15 degrees C) in Atlantic salmon (Salmo salar) cage-farmed in Chile affecting both smolts and adult fish cultured in estuary and marine waters, reaching in some occasions a cumulative mortality up to 25% of the affected population. Diseased fish showed exophthalmia with accumulation of purulent and haemorrhagic fluid around eyes, and ventral petechial haemorrhages. At necropsy, haemorrhage in the abdominal fat, pericarditis, and enlarged liver, spleen and kidney are common pathological changes. Gram-stained smears revealed the presence of Gram-positive cocci, beta-hemolytic, negative for oxidase and catalase tests. Although biochemical characterization of the isolates using the miniaturized system rapid ID 32 Strep suggested their assignation to genus Gemella, sequencing and RFLP analysis of the 16S rRNA revealed that bacteria associated with the mortalities belong to Streptococcus phocae. Serological studies demonstrated that all the salmon isolates are antigenically homogeneous, which can facilitate the development of preventive measures and, although sharing some antigenical determinants, they belong to a different Lancefield group than the type strain isolated from seals. On the basis of these facts, we conclude that the species S. phocae is an emerging pathogen for salmonid culture in Chile, and it should be included as a new member of the warm water streptococcosis.     

Surface properties of Streptococcus phocae strains isolated from diseased Atlantic salmon, Salmo salar L

Streptococcus phocae is an emerging pathogen for Chilean Atlantic salmon, Salmo salar, but the factors determining its virulence are not yet elucidated. In this work, cell surface–related properties such as hydrophobicity and haemagglutination, adhesion to mucus and cell lines, capsule detection, survival and biofilm formation in skin mucus and serum resistance of the isolates responsible for outbreaks in Atlantic salmon and seals were examined. Adhesion to hydrocarbons and the results of salt aggregation tests indicated most of the S. phocae were strongly hydrophobic. All isolates exhibited a similar ability to attach to the Chinook salmon embryo (CHSE) cells line, but were not able to enter CHSE cells. Haemagglutination was not detected. Our data clearly indicate that S. phocae can resist the killing activity of mucus and serum and proliferate in them, which could be associated with the presence of a capsular layer around the cells. Pathogenicity studies using seal and fish isolates demonstrated mortality or pathological signs in fish injected only with the Atlantic salmon isolate. No mortalities or histopathological alterations were observed in fish injected with extracellular products.