Staphylococcus aureus produces membrane-derived vesicles (MVs), which share functional properties to outer membrane vesicles. Atomic force microscopy revealed that S. aureus-derived MVs are associated with the bacterial surface or released into the surrounding environment depending on bacterial growth conditions. By using a comparative proteomic approach, a total of 131 and 617 proteins were identified in MVs isolated from S. aureus grown in Luria-Bertani and brain-heart infusion broth, respectively. Purified S. aureus MVs derived from the bacteria grown in either media induced comparable levels of cytotoxicity and neutrophil-activation. Administration of exogenous MVs increased the resistance of S. aureus to killing by whole blood or purified human neutrophils ex vivo and increased S. aureus survival in vivo. Finally, immunization of mice with S. aureus-derived MVs induced production of IgM, total IgG, IgG1, IgG2a, and IgG2b resulting in protection against subcutaneous and systemic S. aureus infection. Collectively, our results suggest S. aureus MVs can influence bacterial–host interactions during systemic infections and provide protective immunity in murine models of infection.

The aims of this study was to assess the effect of two lactic acid bacteria (LAB), Lactobacillus curvatus and Leuconostoc mesenteroides, originally isolated from gastrointestinal (GI) tract of beluga (Huso huso) and Persian sturgeon (Acipenser persicus), respectively, on growth, survival and digestive enzyme (amylase, lipase and protease) activities and the population level of LAB in the GI tract. The treatments included 10 different groups; control, separate supplements of L. curvatus and Leu. mesenteroides at three different counts [2 · 10 9 , 5 · 10 9 and 9 · 10 9 colony forming units (CFU) per gram food] and three combinations of the two LAB (2 · 10 9 + 2 · 10 9 , 5 · 10 9 + 5 · 10 9 and 9 · 10 9 + 9 · 10 9 CFU per gram food). The bacteria used in this study were added in lyophilized form to chopped Chironomidae. In the beluga study, highest specific growth rate, survival and improved intestinal enzyme activities were noted in the rearing group fed 9 · 10 9 L. curvatus per gram food. In Persian sturgeon, the inclusion level of 2 · 10 9 Leu. mesenteroides had similar positive effect. The ability of LAB to colonize the digestive tract seems to involve host specificity, and our bacteriological results are relevant to initiate future probiotic studies in sturgeons and future directions will be discussed. KEY WORDS

In the present study the intestinal sac method (ex vivo) was used to evaluate the interactions between lactic acid bacteria and staphylococci in the gastrointestinal (GI) tract of beluga (Huso huso). The distal intestine (DI) of beluga was exposed ex vivo to Staphylococcus aureus, Leuconostoc mesenteroides and Lactobacillus plantarum. Histological changes following bacterial exposure were assessed by light and electron microscopy. Control samples and samples exposed only to Leu. mesenteroides and a combination of Leu. mesenteroides and Staph. aureus, had a similar appearance to intact intestinal mucosal epithelium, with no signs of cellular damage. However, exposure of the DI to Staph. aureus and L. plantarum resulted in damaged epithelial cells and disorganized microvilli. Furthermore, 16S rDNA PCR denaturing gradient gel electrophoresis (PCR-DGGE) was used to investigate the adherent microbiota of distal beluga intestine. Several bacterial species were identified by DGGE in the present study that have not previously been identified in beluga.

It is becoming increasingly understood that the current paradigms of in vitro antimicrobial susceptibility testing may have significant shortcomings in predicting activity in vivo. This study evaluated the activity of several antibiotics alone and in combination against clinical isolates of Salmonella enterica serotype Newport (meningitis case) utilizing both conventional and physiological media. In addition, the interactions of these antibiotics with components of the innate immune system were evaluated. Azithromycin, which has performed quite well clinically despite high MICs in conventional media, was shown to be more active in physiological media and to enhance innate immune system killing. Alternatively, chloramphenicol did not show enhanced immune system killing, paralleling its inferior clinical performance to other antibiotics that have been used to treat Salmonella meningitis. These findings are important additions to the building understanding of current in vitro antimicrobial assay limitations that hopefully will amount to future improvements in these assays to better predict clinical efficacy and activity in vivo.

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