Isolation and Genetic Characterization of Streptococcus iniae Virulence Factors in Adriatic Sturgeon (Acipenser naccarii)

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Abstract

The first case of infection of Streptococcus iniae in Adriatic sturgeon (Acipenser naccarii) was recently reported in a raceway system located in Northern Italy. A second episode of infection in sturgeons with absence of mortality and evident clinical signs, was registered in November 2020 in the same farm and is reported in this study. Histopathological changes observed in infected organs are described. The strains isolated in the two episodes were compared using molecular analysis based on PCR, phylogeny and virulence factors analysis. Not all the major virulence factors were detected for the two strains; in particular the strains 78697, isolated in November, lacks cpsD, compared to the strains 64844, isolated in September. Moreover, genetic variations were reported for lctO and pmg genes. These findings let us hypothesize a different virulence of the strains in accordance with clinical findings related to the sturgeons.

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Prospects for inferring very large phylogenies by using the neighbor-joining method.

K Tamura, M Nei, S. Kumar (2004)

Current efforts to reconstruct the tree of life and histories of multigene families demand the inference of phylogenies consisting of thousands of gene sequences. However, for such large data sets even a moderate exploration of the tree space needed to identify the optimal tree is virtually impossible. For these cases the neighbor-joining (NJ) method is frequently used because of its demonstrated accuracy for smaller data sets and its computational speed. As data sets grow, however, the fraction of the tree space examined by the NJ algorithm becomes minuscule. Here, we report the results of our computer simulation for examining the accuracy of NJ trees for inferring very large phylogenies. First we present a likelihood method for the simultaneous estimation of all pairwise distances by using biologically realistic models of nucleotide substitution. Use of this method corrects up to 60% of NJ tree errors. Our simulation results show that the accuracy of NJ trees decline only by approximately 5% when the number of sequences used increases from 32 to 4,096 (128 times) even in the presence of extensive variation in the evolutionary rate among lineages or significant biases in the nucleotide composition and transition/transversion ratio. Our results encourage the use of complex models of nucleotide substitution for estimating evolutionary distances and hint at bright prospects for the application of the NJ and related methods in inferring large phylogenies.

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Streptococcus iniae: an aquatic pathogen of global veterinary significance and a challenging candidate for reliable vaccination.

A. Barnes, A BARNES (2007)

Streptococcus iniae has become one the most serious aquatic pathogens in the last decade causing high losses in farmed marine and freshwater finfish in warmer regions. Although first identified in 1976 from a captive Amazon freshwater dolphin, from which it derives its name, disease outbreaks had most likely been occurring for several decades in marine aquaculture in Japan. S. iniae is globally distributed throughout warm water finfish aquaculture. In common with other encapsulated beta-haemolytic streptococci and in direct contradiction to the phenomenal success story of bacterial vaccines in finfish aquaculture, control of S. iniae by vaccination has met with limited success. Thus, antibiotic usage is the current practice for reducing mortality and consequent economic loss. Vaccine failure appears to result in part from serotypic variation and, whilst 2 serotypes have been named, variation would appear to be more complex. S. iniae also has zoonotic potential, with human infections identified in the USA, Canada, and throughout Asia. In humans, infection is clearly opportunistic with all cases to date associated with direct infection of puncture wounds during preparation of contaminated fish, and generally in elderly or immunocompromised individuals. Significant progress has been made in terms of research into pathogenic mechanisms of S. iniae, with recent research elucidating the role of capsule in virulence for fish through antiopsonic activity. In light of this recent coverage in the literature, the present review centres on areas of direct veterinary interest including identification, epidemiology, therapy and prevention in farmed finfish. Clearly as the prevalence of S. iniae and associated economic losses continue to increase, further work towards developing a reliable vaccine is essential. This would appear to require a much better understanding of cell-surface variability amongst S. iniae isolates.

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Streptococcus iniae phosphoglucomutase is a virulence factor and a target for vaccine development.

John T Buchanan, Jason A Stannard, Xavier Lauth … (2005)

Streptococcus iniae represents a major health and economic problem in fish species worldwide. Random Tn917 mutagenesis and high-throughput screening in a hybrid striped bass (HSB) model of meningoencephalitis identified attenuated S. iniae mutants. The Tn917 insertion in one mutant disrupted an S. iniae homologue of a phosphoglucomutase (pgm) gene. Electron microscopy revealed a decrease in capsule thickness and cell wall rigidity, with DeltaPGM mutant cells reaching sizes approximately 3-fold larger than those of the wild type (WT). The DeltaPGM mutant was cleared more rapidly in HSB blood and was more sensitive to killing by cationic antimicrobial peptides including moronecidin from HSB. In vivo, the DeltaPGM mutant was severely attenuated in HSB, as intraperitoneal challenge with 1,000 times the WT lethal dose produced only 2.5% mortality. Reintroduction of an intact copy of the S. iniae pgm gene on a plasmid vector restored antimicrobial peptide resistance and virulence to the DeltaPGM mutant. In analysis of the aborted infectious process, we found that DeltaPGM mutant organisms initially disseminated to the blood, brain, and spleen but were eliminated by 24 h without end organ damage. Ninety to 100% of fish injected with the DeltaPGM mutant and later challenged with a lethal dose of WT S. iniae survived. We conclude that the pgm gene is required for virulence in S. iniae, playing a role in normal cell wall morphology, surface capsule expression, and resistance to innate immune clearance mechanisms. An S. iniae DeltaPGM mutant is able to stimulate a protective immune response and may have value as a live attenuated vaccine for aquaculture.