Hydrophobic and adhesive patterns of lactic acid bacteria and their antagonism against foodborne pathogens on tomato surface (Solanum lycopersicum L.).

AIMS: To evaluate tomato epiphyte lactic acid bacteria (LAB) hydrophobicity and auto-aggregation as an indicator of bacteria adhesion to tomato. Likewise, use LAB adhesion and co-aggregation as mechanisms to antagonize pathogen attachment. METHODS AND RESULTS: Fifty-four LAB were screened to evaluate their hydrophobic, auto- and co-aggregative properties against Salmonella Typhimurium, Saintpaul, Montevideo and Escherichia coli O157:H7. Subsequently, tomato adhesion of Enterococcus faecium Col1-1C, Weisella cibaria 11-E-2 and W. confusa Col 1-13 with high, medium and low hydrophobicity and high co-aggregation was investigated as well as their pathogen antagonism. Results indicate that bacteria hydrophobicity and auto-aggregation correspond to LAB adhesion to tomato. Enterococcus faecium Col1-1C interfered in most of the pathogen adhesion and micrographs revealed that such effect could be related to the inhibition of structures-type biofilm on E. coli O157:H7 and the aggregate formation on Salmonella. CONCLUSIONS: Lactic acid bacteria hydrophobicity and auto-aggregation can estimate bacteria adhesion to tomato and adhesive and co-aggregative properties could serve as a tool to antagonize foodborne pathogens under specific conditions. SIGNIFICANCE AND IMPACT OF THE STUDY: This study evidence the interference of Ent. faecium Col1-1C in E. coli O157:H7 biofilm formation and Salmonella colonization.

In vivo and in vitro studies using larval and adult antigens from Neobenedenia melleni on immune response in yellowtail (Seriola lalandi).

Neobenedenia melleni is a monogenean parasite that causes significant mortality and economic losses in fish aquaculture. Changes in the antigenic composition of this parasite occur during its developmental stages. In this study, we evaluated humoral parameters in serum and transcriptional immune responses of yellowtail naturally infected with N. melleni. In addition, in vitro assays were performed to study the stimulatory effects of antigens from larvae and adults on spleen leucocytes from non-infected fish at 6 and 24 h post-stimulation. The results showed enhanced total protein, myeloperoxidase and antiprotease activities in N. melleni-infected fish compared with non-infected ones. The induction of Toll-like receptors (TLRs) and pro-inflammatory cytokines in spleen leucocytes during natural infection with N. melleni suggests that these immune-related genes play an important role in the initiation of the immune defence mechanism for controlling parasite infection. Interestingly, the magnitude of in vitro responses of spleen leucocytes was dependent on the parasitic stage. An important stimulation of gene expression by adult antigens on spleen leucocytes was observed. Differential expression patterns of TLRs and target cytokines in yellowtail leucocytes in both in vivo and in vitro studies suggest that the quality of yellowtail immune response is conditioned by N. melleni development.

Effects of immunization of Pelibuey lambs with Oestrus ovis digestive tract protein extracts on larval establishment and development.

Larval midgut proteins of hematophagous parasites contain strong antigens that can be used for host immunization. This concept has been applied for immunization of Pelibuey sheep against Oestrus ovis L. (Diptera: Oestridae). The aim of this study was to examine the effect of immunization on larval establishment (LE) and development. Immunized lambs (I, n = 6) received two injections of crude gut membrane protein extracts (GMPE) from third instar larvae with Freund's incomplete adjuvant (FIA) on days 0 (Day of first immunization) and 21 (0.4 and 0.45 mg GMPE/lamb, respectively). The control group (C, n = 5) received physiological saline with FIA. Lambs were challenged with first instars on Day 29 (20 larvae) and Day 43 (25 larvae). Blood samples were collected biweekly and IgG titers were analyzed by ELISA. All lambs were slaughtered on Day 90 and number of larvae recovered, larval stage and larval weight were recorded at necropsy. No significant effect of immunization on LE (C = 28.9%; I = 31.0% P > 0.05) was observed. Antibody titers were higher in the immunized group on Day 28 (P < 0.05), but subsequently similar in both groups. Larval physiological age and weight were also significantly (P < 0.05) affected by immunization. Immunization of Pelibuey lambs with GMPE did not affect LE but did delay O. ovis larval development.

Ostreid herpesvirus 1 (OsHV-1) detection among three successive generations of Pacific oysters (Crassostrea gigas).

Ostreid Herpesvirus 1 (OsHV-1) was likely detected in Pacific oysters, Crassostrea gigas, at different stages of development. Viral infections were associated with high mortality rates in the spat and larvae. Furthermore, the persistance of OsHV-1 in asymptomatic adults was demonstrated by detection of viral DNA and proteins. In the present study, three successive generations of C. gigas (G0 and G1 parental oysters, G1 and G2 larvae) were screened for OsHV-1 by PCR. Viral DNA was detected in 2-day-old larvae, indicating that infection may take place at very early stages. Although results strengthen the hypothesis of a vertical transmission, it was not possible to predict the issue of a particular type of cross. Indeed, the detection of viral DNA in parental oysters did not systematically correspond to a productive infection or result in a successful transmission to the progeny. However, the infective status of the parents appeared to have an influence on both the infection and the survival rates of the progeny. Crosses involving an OsHV-1 infected male and a non-infected female resulted in hatching and larval survival rates statistically lower than those observed in the other types of cross. These results suggest that OsHV-1-infected females may transmit to their offspring some kind of protection or resistance against viral infection.

Diagnosis of Ostreid herpesvirus 1 in fixed paraffin-embedded archival samples using PCR and in situ hybridisation.

In 1994, some of the high mortality episodes that affected oysters cultured in France were associated with herpesviral infections. Through histology analysis, however, viral presence could only be suspected and confirmation of histological diagnosis by transmission electron microscopy was performed in only a few cases. Subsequently, the characterisation and genome sequencing of Ostreid herpesvirus 1 (OsHV-1) made possible the development of specific molecular detection (PCR and in situ hybridisation (ISH)). Using both molecular tools, attempts were made to screen for OsHV-1 a number of fixed, paraffin-embedded oyster samples collected and processed in 1994. The aim was to compare these techniques and to estimate the accuracy of histology-based indication of viral infection. Existing DNA extraction protocols were adapted for oyster samples and two pairs of specific primers targeting small fragments (less than 200bp) were designed (C(9)/C(10) and B(4)/B(3)). The poor consistency observed between the results of PCR with both primer pairs was confirmed by statistical analysis. C(9)/C(10), which targets a repeated region of the OsHV-1 genome, appears to be the primer of choice for viral detection in archival samples. In situ hybridisation may furnish complementary information concerning the localisation of viral foci. Under certain conditions, retrospective examination of archival samples by molecular techniques may therefore provide valuable epidemiological data.

Comparative susceptibility of veliger larvae of four bivalve mollusks to a Vibrio alginolyticus strain.

The susceptibility of 7 d old veliger larvae of the scallops Argopecten ventricosus and Nodipecten subnodosus, the penshell Atrina maura, and the Pacific oyster Crassostrea gigas to a pathogenic strain of Vibrio alginolyticus was investigated by challenging the larvae with different bacterial concentrations in a semi-static assay. The results indicate that the larvae of the 2 scallop species are more susceptible to the V. alginolyticus strain than those of the oyster and the penshell. Signs of the disease were similar to bacillary necrosis described in previous work. Interspecies differences in susceptibility to pathogens are discussed.

Effects of different stressors in haematological variables in cultured Oreochromis aureus S.

Since haematological variables can be used to assess the health state in cultured fish, a haematological characterization of clinically healthy Oreochromis aureus was done to establish the reference indices of this species. Fish were subjected to different stressed conditions (bacterial infection, nitrite intoxication, malachite green overdose) to study the changes in the haematological indices and its relation with the health condition. This species showed microcytic anaemia under experimental bacterial infection by Corynebacterium sp.; anaemia, neutrophilia and erythrocytes deformation following nitrite intoxication and medication overdose with malachite green.

Cell yield and superoxide dismutase activity of the marine yeast Debaryomyces hansenii under different culture conditions.

The effect of aeration, pH, stirring rate, and temperature on the biomass production and superoxide dismutase (SOD) activity of the marine yeast Debaryomyces hansenii strain C-11 was determined. The cell biomass yield was approximately 50% in a seawater-formulated medium using glucose as the carbon source. The SOD activity increased by application of a pulse of oxygen or 0.8 mM sulfate copper into the chemical reactor. The SOD enzyme had an activity of 400 units/mg of protein in a crude extract produced under such conditions, the best activity ever reported for this enzyme in a crude preparation.