Evaluation of characteristics of Pediococcus spp. to be used as a vaginal probiotic

Aims

The aim of our research was to select, identify and characterize an isolate of lactic acid bacteria to be considered as a vaginal probiotic.

Methods and Results

Thirty‐five isolates of Pediococcus spp. showed bacteriocinogenic activity against Listeria monocytogenes and the ability to survive in simulated vaginal fluid (SVF) at pH 4·2. One isolate of Pediococcus spp. was selected and characterized to evaluate its safety before the use as vaginal probiotic. Pediococcus pentosaceus SB83 did not show the presence of virulence factors such as the production of gelatinase, lipase and DNase, haemolytic activity, nor the presence of virulence genes (genes esp, agg, gelE, efaAfm, efaAfs, cylA, cylB and cylM). Pediococcus pentosaceus SB83 was considered sensitive to chloramphenicol, gentamicin, streptomycin, kanamycin, erythromycin and ampicillin. This strain was considered resistant to tetracycline and vancomycin. Pediococcus pentosaceus SB83 was a biofilm producer at different pH values (4·2, 5·5 and 6·5) in SVF and in de Man, Rogosa and Sharpe medium.

Conclusions

The in vitro results provide a basis for the use of P. pentosaceus SB83 as a vaginal probiotic, to prevent colonization by L. monocytogenes in pregnant women.

Significance and Impact of Study

The application of vaginal probiotics could have the potential for preventing vaginal infections and consequently reduce abortion and neonatal infections.     

Comparison of biological and demographic parameters forMononychellus progresivus andOligonychus gossypii on cassava: influence of temperature

The effect of five constant temperatures (16, 22, 26, 31 and 36°C) on biological (survival and duration of developmental stages, fecundity and longevity of females, sex-ratio) and demographic parameters (R _o , G, r _m and λ) of the two main phytophagous mites that attack cassava in Africa,Mononychellus progresivus Doreste andOligonychus gossypii (Zacher), was studied in the laboratory. Experiments were performed simultaneously on the two mite species reared on the same cassava variety (1M20) under controlled conditions: 70±10% r.h. and 12L∶12D. The lower thermal threshold was 13°C forM. progresivus and 11°C forO. gossypii. Both species developed in the range from 22 to 36°C. The shortest development time was obtained at 31°C for both species; it was 7.2 days forM. progresivus and 8.2 days forO. gossypii. Maximum fecundity of both species was recorded at 26°C with 42.1 eggs forM. progresivus and 36.3 eggs forO. gossypii The highest intrinsic rate of increase (r _m ) was obtained at 31°C for both species with 0.289 and 0.214 forM. progresivus andO. gossypii, respectively.     

Frequency, type, and distribution of EST-SSRs from three genotypes of Lolium perenne, and their conservation across orthologous sequences of Festuca arundinacea, Brachypodium distachyon, and Oryza sativa

Background  

Simple sequence repeat (SSR) markers are highly informative and widely used for genetic and breeding studies in several plant species. They are used for cultivar identification, variety protection, as anchor markers in genetic mapping, and in marker-assisted breeding. Currently, a limited number of SSR markers are publicly available for perennial ryegrass (Lolium perenne). We report on the exploitation of a comprehensive EST collection in L. perenne for SSR identification. The objectives of this study were 1) to analyse the frequency, type, and distribution of SSR motifs in ESTs derived from three genotypes of L. perenne, 2) to perform a comparative analysis of SSR motif polymorphisms between allelic sequences, 3) to conduct a comparative analysis of SSR motif polymorphisms between orthologous sequences of L. perenne, Festuca arundinacea, Brachypodium distachyon, and O. sativa, 4) to identify functionally associated EST-SSR markers for application in comparative genomics and breeding.     

A family history of DUX4: phylogenetic analysis of DUXA, B, C and Duxbl reveals the ancestral DUX gene

Background  

DUX4 is causally involved in the molecular pathogenesis of the neuromuscular disorder facioscapulohumeral muscular dystrophy (FSHD). It has previously been proposed to have arisen by retrotransposition of DUXC, one of four known intron-containing DUX genes. Here, we investigate the evolutionary history of this multi-member double-homeobox gene family in eutherian mammals.     

Effect of iron on the expression of sirR and sitABC in biofilm-associated Staphylococcus epidermidis

Background  

Different gene expression patterns correlate with the altered phenotype in biofilm-associated bacteria. Iron and iron-linked genes are thought to play a key-role in biofilm formation. The expression of Fe-linked genes (sirR, sitABC operon) in Staphylococcus epidermidis, was compared in planktonic versus sessile bacteria in vitro and in vivo in a subcutaneous foreign body rat model.     

First multi-locus sequence typing scheme for Arcobacter spp.

Background  

Arcobacter spp. are a common contaminant of food and water, and some species, primarily A. butzleri and A. cryaerophilus, have been isolated increasingly from human diarrheal stool samples. Here, we describe the first Arcobacter multilocus sequence typing (MLST) method for A. butzleri, A. cryaerophilus, A. skirrowii, A. cibarius and A. thereius.     

Interaction ofManduca sexta resistance in tomato with insect predators ofHelicoverpa zea

Resistance in accession PI 134417 ofLycopersicon hirsutum f.glabratum toManduca sexta L. results from the presence of the methyl ketones 2-tridecanone and 2-undecanone in the type VI trichome glands on foliage of these plants. Short (24 h) and long term (neonate to adult) laboratory experiments usingM. sexta-resistant and susceptible foliage and methyl ketone treated filter paper disks were conducted to evaluate the direct (plant-mediated), and indirect (prey-mediated) effects of this resistance on two predators ofHelicoverpa zea eggs,Coleomegilla maculata (DeGeer) andGeocoris punctipes (Say). Direct effects of resistance were manifest as reduced egg consumption and increased mortality for both predators. Indirect effects were manifest as reduced egg consumption byG. punctipes, but were not observed forC. maculata. Results of experiments using methyl ketone-treated filter paper disks instead of foliage were similar.     

Paraphyly of organelle DNAs in Cycas Sect. Asiorientale s due to ancient ancestral polymorphisms

Background  

This study addresses the apportionment of genetic diversity between Cycas revoluta and C. taitungensis, species that constitute the section Asiorientales and represent a unique, basal lineage of the Laurasian genus Cycas. Fossil evidence indicates divergence of the section from the rest of Cycas at least 30 million years ago. Geographically, C. taitungensis is limited to Taiwan whereas C. revoluta is found in the Ryukyu Archipelago and on mainland China.     

The gene transformer-2 of Sciara (Diptera, Nematocera) and its effect on Drosophila sexual development

Background  

The gene transformer-2, which is involved in sex determination, has been studied in Drosophila, Musca, Ceratitis, Anastrepha and Lucilia. All these members of Diptera belong to the suborder Brachycera. In this work, it is reported the isolation and characterisation of genes transformer-2 of the dipterans Sciara ocellaris and Bradysia coprophila (formerly Sciara coprophila), which belong to the much less extensively analysed Sciaridae Family of the Suborder Nematocera, which is paraphyletic with respect to Suborder Brachycera.     

Tracking costs of virulence in natural populations of the wheat pathogen, Puccinia striiformis f.sp. tritici

Background  

Costs of adaptation play an important role in host-parasite coevolution. For parasites, evolving the ability to circumvent host resistance may trade off with subsequent growth or transmission. Such costs of virulence (sensu plant pathology) limit the spread of all-infectious genotypes and thus facilitate the maintenance of genetic polymorphism in both host and parasite. We investigated costs of three virulence factors in Puccinia striiformis f.sp. tritici, a fungal pathogen of wheat (Triticum aestivum).     

Efficacy ofCarpophilus aggregation pheromones on nine species in northeastern Ohio, and identification of the pheromone ofC. brachypterus

Aggregation pheromones for sevenCarpophilus (Coleoptera: Nitidulidae) species were field tested at a site with a rich nitidulid fauna in Ohio, USA, during the summers of 1992 and 1993. The pheromones used were blends identified for:Carpophilus antiquus (Melsheimer),C. brachypterus (Say),C. freemani Dobson, C. hemipterus (L.),C. lugubris Murray,C. mutilatus Erichson, andC. obsoletus Erichson. Each pheromone was used in conjunction with whole wheat bread dough, an effective co-attractant. The pheromone ofC. brachypterus Say was identified during the course of this study and was also tested; males emitted a 100:6:11:4:3 blend of (2E, 4E, 6E, 8E)-3, 5, 7-trimethyl-2, 4, 6, 8-decatetraene, (2E, 4E, 6E, 8E)-3, 5, 7-trimethyl-2, 4, 6, 8-undecatetraene, (2E, 4E, 6E, 8E)-7-ethyl-3, 5-dimethyl-2, 4, 6, 8-decatetraene, (3E, 5E, 7E, 9E)-4, 6, 8-trimethyl-3, 5, 7, 9-undecatetraene and (2E, 4E, 6E, 8E)-7-ethyl-3, 5-dimethyl-2, 4, 6, 8-undecatetraene, respectively. All species responded favorably to their own pheromones with the exception ofC. obsoletus, which was not present in this area. Strong mutual cross attraction was observed betweenC. brachypterus andC. hemipterus. In addition,C. lugubris responded to the pheromones ofC. obsoletus andC. hemipterus and, more weakly, to those ofC. freemani andC. brachypterus; C. freemani responded slightly to the pheromone ofC. multilatus; andC. antiquus responded to the pheromone ofC. lugubris. In most cases, cross attraction was related to the species involved sharing pheromone components. ForC. antiquus, however, the response to theC. lugubris pheromone was apparently kairomonalC. corticinus, C. marginatus, C. marginellus, andC. sayi, for which pheromones are not known, were attracted to the pheromone ofC. lugubris and in some cases to other pheromones. Significant numbers ofColopterus spp. responded to the blends forC. lugubris, C. hemipterus, C. brachypterus, and probably,C. obsoletus.     

New probes used for IS1245 and IS1311 restriction fragment length polymorphism of Mycobacterium avium subsp. avium and Mycobacterium avium subsp. hominissuis isolates of human and animal origin in Norway

Background  

Mycobacterium avium is an environmental mycobacterium that can be divided into the subspecies avium, hominissuis, paratuberculosis and silvaticum. Some M. avium subspecies are opportunistic pathogens for animals and humans. They are ubiquitous in nature and can be isolated from natural sources of water, soil, plants and bedding material. Isolates of M. avium originating from humans (n = 37), pigs (n = 51) and wild birds (n = 10) in Norway were examined by IS1245 and IS1311 RFLP using new and specific probes and for the presence of IS901 and ISMpa1 by PCR. Analysis and generation of a dendrogram were performed with the software BioNumerics.     

Improving Salmonella vector with rec mutation to stabilize the DNA cargoes

Background  

Salmonella has been employed to deliver therapeutic molecules against cancer and infectious diseases. As the carrier for target gene(s), the cargo plasmid should be stable in the bacterial vector. Plasmid recombination has been reduced in E. coli by mutating several genes including the recA, recE, recF and recJ. However, to our knowledge, there have been no published studies of the effect of these or any other genes that play a role in plasmid recombination in Salmonella enterica.     

Paracoccidioides brasiliensis pancreatic destruction in Calomys callosus experimentally infected

Background  

The wild rodent Calomys callosus is notably resistant to Trypanosoma cruzi infection. In order to better characterize this animal model for experimental infections, we inoculated C. callosus intraperitoneally with Paracoccidioides brasiliensis, a thermally dimorphic fungus that causes a chronic disease with severe granuloma formation in the mouse and humans. The dissemination of P. brasiliensis cells through the lungs, liver, pancreas, and spleen was assessed by histological analysis.     

Allelic diversity and phylogeny of homB, a novel co-virulence marker of Helicobacter pylori

Background  

The homB gene is a Helicobacter pylori disease-marker candidate, strongly associated with peptic ulcer disease, while homA, its paralogue gene with 90% sequence identity, is correlated with non-ulcer dyspepsia. The HomB encoded outer membrane protein was shown to contribute to the proinflammatory properties of H. pylori and also to be involved in bacterial adherence.     

Effects of genetic and environmental host plant variation on the susceptibility of two noctuids toBacillus thuringiensis

TwoApium graveolens var.rapaceum (L.) cultivars that differ in their suitability for the survival and growth ofSpodoptera exigua (Hübner) andTrichoplusia ni (Hübner) were used to examine the effect of genetic and seasonal environmental variation in host plant suitability on the efficacy ofBacillus thuringiensis subsp.kurstaki (Berliner). The effects of host plant genotype andB. thuringiensis were generally independent, so thatB. thuringiensis efficacy was greatest on the resistant host plant cultivar. Host plant suitability varied within growing season for both insect species but, while host plant suitability decreased with increasing plant age forT. ni, the response ofS. exigua to plant age was not as clear. Within season variation in host plant suitability affectedB. thuringiensis efficacy and the interaction betweenB. thuringiensis and host plant cultivar forS. exigua but not forT. ni. Soluble protein and Folin-Denis phenolic concentrations of host plant tissue were not correlated with changes in host plant suitability to either insect species.     

AI-2 does not function as a quorum sensing molecule inCampylobacter jejuniduring exponential growthin vitro

Background  

Campylobacter jejunicontains a homologue of theluxSgene shown to be responsible for the production of the signalling molecule autoinducer-2 (AI-2) inVibrio harveyiandVibrio cholerae. The aim of this study was to determine whether AI-2 acted as a diffusible quorum sensing signal controllingC. jejunigene expression when it is produced at high levels during mid exponential growth phase.     

Genome constitutions of Pseudoroegneriageniculata, P. geniculata ssp. scythica and P. geniculata ssp. pruinifera (Poaceae: Triticeae) revealed by genomic in situ hybridization

Genomic constitutions of three taxa of Pseudoroegneria á. L?ve, P. geniculata, P. geniculata ssp. scythica, and P. geniculata ssp. pruinifera, were characterized using genomic in situ hybridization (GISH). The results indicated that ploidy level and genomic constitution in the three Pseudoroegneria taxa studied were as follows: P. geniculata (2n = 4x = 28; genomic formula StSt ^g ), P. geniculata ssp. scythica (2n = 4x = 28; genomic formula ESt), and P. geniculata ssp. pruinifera (2n = 6x = 42; genomic formula EESt). P. geniculata ssp. scythica and P. geniculata ssp. pruinifera should be transferred to Trichopyrum á. L?ve following L?ve’s principles and treated as T. scythicum and T. pruiniferum, respectively.     

qPCR in gastrointestinal stromal tumors: Evaluation of reference genes and expression analysis of KIT and the alternative receptor tyrosine kinases FLT3, CSF1-R, PDGFRB, MET and AXL

Background  

Gastrointestinal stromal tumors (GIST) represent the most common mesenchymal tumors of the gastrointestinal tract. About 85% carry an activating mutation in the KIT or PDGFRA gene. Approximately 10% of GIST are so-called wild type GIST (wt-GIST) without mutations in the hot spots. In the present study we evaluated appropriate reference genes for the expression analysis of formalin-fixed, paraffin-embedded and fresh frozen samples from gastrointestinal stromal tumors. We evaluated the gene expression of KIT as well as of the alternative receptor tyrosine kinase genes FLT3, CSF1-R, PDGFRB, AXL and MET by qPCR. wt-GIST were compared to samples with mutations in KIT exon 9 and 11 and PDGFRA exon 18 in order to evaluate whether overexpression of these alternative RTK might contribute to the pathogenesis of wt-GIST.