Abstract: The most demanded organic foods worldwide are those that are consumed fresh, such as fruits and vegetables. However, there is a knowledge gap about some aspects of organic food microbiological quality and safety. Organic fruits and vegetables are more exposed to pathogenic microorganisms due to surface contact with natural fertilizers such as animal manure, wastes and vermicompost used during farming. Therefore, the objective of this work was to study the contamination of organic fresh green leafy vegetables by two emergent pathogens, Arcobacter spp. and Helicobacter pylori. For this purpose, a total of 24 vegetable samples, 13 lettuce and 11 spinach were acquired from 10 different ecological supermarkets and greengroceries and analyzed by culture and PCR. Arcobacter spp. was detected in five samples (20%) by PCR, four spinach and one lettuce. One spinach sample was found to be also positive by culture. For H. pylori, the H. pylori VacA gene-specific band was detected in 12 vegetable samples (50%), 10 lettuces and two spinach. Isolation in the selective medium did not yield any positive result, possibly because of low contamination levels together with the presence of the organism in its viable but non-culturable form. Results showed significant levels of H. pylori and Arcobacter contamination in organic vegetables that are generally consumed raw, which seems to confirm that these foods can act as transmission vehicles to humans.
Abstract: Polymerase chain reaction (PCR) assays targeting genomic DNA segments have been established for the detection of Helicobacter pylori in clinical specimens. However, the data on comparative evaluations of various targets in detection of H. pylori are limited. Furthermore, the frequencies of vacA (s1 and s2) and cagA genotypes, which are suggested to be involved in the pathogenesis of H. pylori in other parts of the world, are not well studied in Kuwait. The aim of this study was to evaluate PCR assays for the detection and genotyping of H. pylori by targeting the amplification of DNA targets from four genomic segments. The genomic DNA were isolated from 72 clinical isolates of H. pylori and tested in PCR with four pairs of oligonucleotides primers, i.e. ECH-U/ECH-L, ET-5U/ET-5L, CagAF/CagAR and Vac1F/Vac1XR, which were expected to amplify targets of various sizes (471 bp, 230 bp, 183 bp and 176/203 bp, respectively) from the genomic DNA of H. pylori. The PCR-amplified DNA were analyzed by agarose gel electrophoresis. PCR products of expected size were obtained with all primer pairs by using genomic DNA isolated from H. pylori. DNA dilution experiments showed that the most sensitive PCR target was 471 bp DNA amplified by the primers ECH-U/ECH-L, followed by the targets of Vac1F/Vac1XR (176 bp/203 DNA), CagAF/CagAR (183 bp DNA) and ET-5U/ET-5L (230 bp DNA). However, when tested with undiluted genomic DNA isolated from single colonies of all isolates, the Vac1F/Vac1XR target provided the maximum positive results (71/72 (99% positives)), followed by ECH-U/ECH-L (69/72 (93% positives)), ET-5U/ET-5L (51/72 (71% positives)) and CagAF/CagAR (26/72 (46% positives)). The results of genotyping experiments showed that vacA s1 (46% positive) and vacA s2 (54% positive) genotypes were almost equally associated with VaCA+/CagA- isolates (P > 0.05), but with VacA+/CagA+ isolates, S1 genotype (92% positive) was more frequently detected than S2 genotype (8% positive) (P< 0.0001). In conclusion, among the primer pairs tested, Vac1F/Vac1XR provided the best results for detection of H. pylori. The genotyping experiments showed that vacA s1 and vacA s2 genotypes were almost equally associated with vaCA+/cagA- isolates, but vacA s1 genotype had a significantly increased association with vacA+/cagA+ isolates.
Abstract: The management of Helicobacter pylori (H. pylori)
eradication is still a matter of discussion, full effectiveness is rarely
achieved, and it has many adverse effects. The use of probiotics may
be associated with better eradication rates and possibly prevention of
adverse events due to antibiotic therapy. The present clinical study
was undertaken to evaluate the efficacy of a specially designed
fermented milk product, containing Bifidobacterium lactis B420, on
the eradication of H. pylori infection in a prospective, randomized,
double-blind, controlled study in humans. Four test fermented milks
(FM) were specially designed in which counts of viable cells in all
products were 10^10 Log CFU. 100 mL-1 for Bifidobacterium lactis -
Bifidobacterium species 420. 190 subjects infected with H. pylori,
with previous diagnosis of functional dyspepsia according to Rome
III criteria entered the study. Bifidobacterium lactis B420,
administered twice a day for 90 days was not able to eradicate H.
pylori in Brazilian patients with functional dyspepsia.
Abstract: Probiotic bacteria especially Lactobacillus spp. and Bifidobacterium exert suppressive effect on Helicobacter pylori. Cinnamon and licorice have been traditionally used for the treatment of gastric ulcer. The objectives of this study were to determine the effects of herbs on yogurt fermentation, the level of probiotic bacteria in yogurt during 28 days storage and the effect of herbal yogurt on the growth of H. pylori in vitro. Cinnamon or licorice was mixed with milk and the mixture was fermented with probiotic bacteria to form herbal-yogurt. Changes of pH and total titratable acids were monitored and the viability of probiotic bacteria was evaluated during and after refrigerated storage. The in vitro inhibition of H. pylori growth was determined using agar diffusion and minimum inhibitory concentration (MIC) method. The presence of herbs did not affect the probiotic population during storage. There were no significant differences in pH and TTA between herbal-yogurts and plain-yogurt during fermentation and storage. Water extract of cinnamon-yogurt showed the highest inhibition effect (13.5mm) on H. pylori growth in comparison with licorice-yogurt (11.2mm). The present findings indicate cinnamon and licorice has bioactive components to decrease the growth of H. pylori.
Abstract: Bacterial molecular chaperone DnaK plays an essential role in protein folding, stress response and transmembrane targeting of proteins. DnaKs from many bacterial species, including Escherichia coli, Salmonella typhimurium and Haemophilus infleunzae are the molecular targets for the insect-derived antimicrobial peptide pyrrhocoricin. Pyrrhocoricin-like peptides bind in the substrate recognition tunnel. Despite the high degree of crossspecies sequence conservation in the substrate-binding tunnel, some bacteria are not sensitive to pyrrhocoricin. This work addresses the molecular mechanism of resistance of Helicobacter pylori DnaK to pyrrhocoricin. Homology modelling, structural and sequence analysis identify a single aminoacid substitution at the interface between the lid and the β-sandwich subdomains of the DnaK substrate-binding domain as the major determinant for its resistance.