Nowadays, the oral use of probiotics is widespread. However, the safety profile with the use of live probiotics is still a matter of debate. Main risks include: Cases of systemic infections due to translocation, particularly in vulnerable patients and pediatric populations; acquisition of antibiotic resistance genes; or interference with gut colonization in neonates. To avoid these risks, there is an increasing interest in non-viable microorganisms or microbial cell extracts to be used as probiotics, mainly heat-killed (including tyndallized) probiotic bacteria (lactic acid bacteria and bifidobacteria). Heat-treated probiotic cells, cell-free supernatants, and purified key components are able to confer beneficial effects, mainly immunomodulatory effects, protection against enteropathogens, and maintenance of intestinal barrier integrity. At the clinical level, products containing tyndallized probiotic strains have had a role in gastrointestinal diseases, including bloating and infantile coli—in combination with mucosal protectors—and diarrhea. Heat-inactivated probiotics could also have a role in the management of dermatological or respiratory allergic diseases. The reviewed data indicate that heat-killed bacteria or their fractions or purified components have key probiotic effects, with advantages versus live probiotics (mainly their safety profile), positioning them as interesting strategies for the management of common prevalent conditions in a wide variety of patients´ characteristics.

Erwinia amylovora, a Gram negative bacteria of the Enterobacteriaceae family, is the causal agent of fire blight, a devastating plant disease affecting a wide range of host species within Rosaceae and a major global threat to commercial apple and pear production. Among the limited number of control options currently available, prophylactic application of antibiotics during the bloom period appears the most effective. Pathogen cells enter plants through the nectarthodes of flowers and other natural openings, such as wounds, and are capable of rapid movement within plants and the establishment of systemic infections. Many virulence determinants of E. amylovora have been characterized, including the Type III secretion system (T3SS), the exopolysaccharide (EPS) amylovoran, biofilm formation, and motility. To successfully establish an infection, E. amylovora uses a complex regulatory network to sense the relevant environmental signals and coordinate the expression of early and late stage virulence factors involving two component signal transduction systems, bis-(3′-5′)-cyclic di-GMP (c-di-GMP) and quorum sensing. The LPS biosynthetic gene cluster is one of the relatively few genetic differences observed between Rubus- and Spiraeoideae-infecting genotypes of E. amylovora. Other differential factors, such as the presence and composition of an integrative conjugative element associated with the Hrp T3SS (hrp genes encoding the T3SS apparatus), have been recently described. In the present review, we present the recent findings on virulence factors research, focusing on their role in bacterial pathogenesis and indicating other virulence factors that deserve future research to characterize them.

Although Anoxybacillus and Geobacillus, two genera of thermophilic bacteria close to the genus Bacillus, have only been described recently, the number of species in these genera has increased rapidly. Four thermophilic, lipolytic strains (DR01, DR02, DR03 and DR04) isolated from a hot spring in Veracruz (Mexico), which could not be identified phenotypically, were subjected to 16S rRNA gene sequence analysis. Three strains were identified as belonging to the genus Anoxybacillus, but strain DR03 was identified as Geobacillus pallidus. This result led us to perform a phylogenetic analysis of the genera Anoxybacillus and Geobacillus based on 16S rRNA gene sequences from all the type strains of these genera. Phylogenetic trees showed three major clusters, Anoxybacillus–Geobacillus tepidamans, Geobacillus sensu stricto and Geobacillus pallidus, while the 16S rRNA gene sequences of G. pallidus (DR03 and the type strain) showed low similarity to sequences of Anoxybacillus (92.5–95.1 %) and Geobacillus (92.8–94.5 %) species, as well as to Bacillus subtilis (92.2–92.4 %). In addition, G. pallidus could be differentiated from Anoxybacillus and Geobacillus on the basis of DNA G+C content and fatty acid and polar lipid profiles. From these results, it is proposed that Geobacillus pallidus should be classified in a novel genus, for which we propose the name Aeribacillus, as Aeribacillus pallidus gen. nov., comb. nov. The type strain of Aeribacillus pallidus is H12T (=ATCC 51176T =DSM 3670T =LMG 19006T).

A polyphasic study was performed to determine the taxonomic position of two Aeromonas strains, 665N and 868E T , isolated from bivalve molluscs, that could not be identified at the species level in a previous numerical taxonomy study. The DNA G+C content of these isolates was 62.3 and 62.6 mol%, respectively. Sequence analysis of the 16S rRNA gene showed that the two new strains were closely related to members of the genus Aeromonas. Fluorescence amplified fragment length polymorphism fingerprinting revealed that strains 665N and 868E T clustered together with a similarity of 78 % but did not cluster with any of the Aeromonas genomospecies. DNA-DNA hybridization experiments revealed a high level of relatedness between the two new isolates (76 %) but low levels of relatedness between these and phylogenetically most closely related Aeromonas genomospecies (30-44 %). Useful tests for the phenotypic differentiation of strains 665N and 868E T from other mesophilic Aeromonas species included those for gas from glucose, lysine decarboxylase, Voges-Proskauer reaction, acid from L-arabinose, hydrolysis of aesculin and utilization of L-lactate. On the basis of genotypic and phenotypic evidence, strains 665N and 868E T are considered to represent a novel species of the genus Aeromonas, for which the name Aeromonas bivalvium sp. nov. is proposed. The type strain is 868E T (=CECT 7113 T =LMG 23376 T ).Members of the genus Aeromonas, belonging to the class Gammaproteobacteria, are Gram-negative, non-spore-forming bacilli or coccobacilli, and are facultatively anaerobic, chemo-organotrophic, oxidase-and catalase-positive, resistant to the vibriostatic agent O/129 (2,4-diamino-6,7-diisopropylpteridine), generally motile by means of a single polar flagellum and are able to reduce nitrate to nitrite. Aeromonads are primarily aquatic, widespread in environmental habitats, frequently isolated from foods and often associated with aquatic animals, and some species are primary or opportunistic pathogens in invertebrates and vertebrates including humans (Martin-Carnahan & Joseph, 2005).At the time of writing, 17 Aeromonas species and 20 DNA-DNA hybridization groups (HGs) have been described: Aeromonas hydrophila (HG1), A. bestiarum (HG2), A. salmonicida (HG3), A. caviae (HG4), A. media (HG5), A. eucrenophila (HG6), A. sobria (HG7), A. veronii bv. Sobria (HG8/10), A. jandaei (HG9), A. veronii bv. Veronii (HG10/ 8), Aeromonas sp. HG11, A. schubertii (HG12), Aeromonas sp. group 501 (HG13), A. trota (HG14), A. allosaccharophila (HG15), A. encheleia (HG16), A. popoffii (HG17), A. culicicola (HG18), A. simiae (HG19) and A. molluscorum (HG20) (Pidiyar et al., 2002; Harf-Monteil et al., 2004; Miñana-Galbis et al., 2004;Martin-Carnahan & Joseph, 2005). In addition to the continuous description of novel species, the complexity of Aeromonas taxonomy results from the isolation of motile and non-motile, mesophilic and psychrophilic, pigmented and non-pigmented strains within several Aeromonas species (Altwegg et al., 1990;Martin-Carnahan & Joseph,...

An analysis of the universal target (UT) sequence from the cpn60 gene was performed in order to evaluate its usefulness in phylogenetic and taxonomic studies and as an identification marker for the genus Aeromonas. Sequences of 555 bp, corresponding to the UT region, were obtained from a collection of 35 strains representing all of the species and subspecies of Aeromonas. From the analysis of these sequences, a range of divergence of 0-23.3 % was obtained, with a mean of 11.2±0.9 %. Comparative analyses between cpn60 and gyrB, rpoD and 16S rRNA gene sequences were carried out from the same Aeromonas strain collection. Sequences of the cpn60 UT region showed similar discriminatory power to gyrB and rpoD sequences. The phylogenetic relationships inferred from cpn60 sequence distances indicated an excellent correlation with the present affiliation of Aeromonas species with the exception of Aeromonas hydrophila subsp. dhakensis, which appeared in a separate phylogenetic line, and Aeromonas sharmana, which exhibited a very loose phylogenetic relationship to the genus Aeromonas. Sequencing of cpn60 from 33 additional Aeromonas strains also allowed us to establish intra-and interspecific threshold values. Intraspecific divergence rates were ¡3.5 %, while interspecific divergence rates fell between 3.7 and 16.9 %, excluding A. sharmana. In this study, cpn60 UT sequencing was shown to be a universal, useful, simple and rapid method for the identification and phylogenetic affiliation of Aeromonas strains.