Occurrence of mycotoxins and microbial communities in artisanal infant flours marketed in Côte d'Ivoire.

The aim of this study was to determine the microbial diversity and mycotoxin profile of artisanal infant flours commonly vended in public healthcare centres and retail markets in Côte d'Ivoire. Thus, maize, millet, sorghum, soya and multigrain (mix of different cereals) flour samples collected from different localities were first, analysed for nutritional composition, then for microbial communities using high-throughput sequencing and for mycotoxins through UHPLC-MS/MS method. Firmicutes was the most abundant bacterial phylum and the dominant genera were Weissella, Staphylococcus, Pediococcus. Potential pathogenic genera such as Bacillus, Enterobacter, Acinetobacter and Burkholderia were also found. The fungal community was composed of two dominant phyla (Ascomycota and Basidiomycota) and 31 genera with > 0.1% relative abundance. In samples from public healthcare centres, Candida, Hyphopichia, Trichosporon, and Cyberlindnera were the most dominant genera according to the flour type while in samples from retail markets, they were Cyberlindnera, Clavispora, Nakaseomyces, Aureobasidium and Candida. Possible toxigenic genera Fusarium and Aspergillus were also detected. Aflatoxin B1 (AFB1), Ochractoxin (OTA), Fumonisin B1 (FB1) and B2 (FB2) were the mycotoxins found in the analysed flours. AFB1 was detected in 100% of maize (range 1.2-120.5 µg/kg; mean: 44.2 µg/kg) and 50-83.3% of millet flours (range 0.2-31.5 µg/kg; mean: 31.5 µg/kg). Its level in all maize and rice flour samples exceeded EU standard (0.1 µg/kg). For OTA and fumonisins, millet and maize flours showed the highest levels of sample exceeding the EU standard. Thus, artisanal infant flours marketed in Côte d'Ivoire, mainly maize and rice flours, although containing potentially beneficial bacteria, represent potential health risks for children.

Occurrence of biogenic amines and their correlation with bacterial communities in the Ivorian traditional fermented fish adjuevan during the storage.

Adjuevan is an Ivorian traditional fermented fish used as a condiment. However, the fermentation process and storage conditions may lead to the production of biogenic amines (BA) which can induce severe human toxicological effects. Thus, this study aimed to reveal the bacterial community diversity and the BA contents during the storage. Samples of adjuevan from the fish species Chloroscombrus chrysurus, Galeoides decadactylus, and Thunnus thynnus were collected from local producers, stored at ambient temperature (28-30 °C) and in a refrigerator (4 °C) over a period of 8 weeks. At 2-week intervals, BA were determined by HPLC and the bacterial communities analyzed using high-throughput sequencing (NGS) of the V3-V4 region of the 16S rRNA gene. Results showed that histamine, cadaverine, putrescine, and tyramine were the major compounds. In adjuevan from T. thynnus, the level of histamine was over the maximum level of 200 mg/kg determined by Codex Alimentarius. For the other amines, no safety concerns are related. In total, 21 bacterial genera with a relative abundance ≥ 1% and belonging to 14 families and 5 phyla were detected. The Bacillaceae family was the most found at ambient temperature while Staphylococcaceae and Enterococcaceae were the most abundant in a refrigerator. The analysis of correlation showed that the increase of Lentibacillus leads to a decrease of the major BA at ambient temperature. On the contrary, the increase of Staphylococcus, Lactobacillus, Psychrobacter, Peptostreptococcus, and Fusobacterium leads to an increase of these biogenic compounds. Thus, Lentibacillus acted as BA-oxidizing bacteria while the others were found as BA-producing bacteria during adjuevan storage.

Impact of turning, pod storage and fermentation time on microbial ecology and volatile composition of cocoa beans.

Cocoa quality depends on several parameters, such as cocoa variety, environmental growth conditions, cultivation technique, and post-harvest treatments applied to coca beans. In this work, we studied the impact of cocoa post-harvest processing on both microbial communities structure and volatile composition. Cocoa beans samples were fermented in wooden boxes in Ivory Coast at different time intervals with turning and without turning, and derived from pods stored for two different duration times. Cocoa beans were analyzed using a molecular fingerprinting method (PCR-DGGE) in order to detect variations in microbial communities' structure; this global analysis was coupled to SPME-GC-MS for assessing cocoa volatile profiles. The results showed that the main parameter that influenced microbial communities structure was fermentation time, followed by turning, whereas, pods storage duration had a minor impact. Similar results were obtained for aromatic profile, except for pods storage duration that significantly affected volatile compound production. Global statistical analysis using Canonical Correspondence Analysis (CCA), showed the relationship between microbial communities and volatile composition. Furthermore, this study allowed the identification of discriminating microbial and chemical markers of cocoa post-harvest processing.

The population structure of Ochrobactrum isolated from entomopathogenic nematodes indicates interactions with the symbiotic system.

Entomopathogenic nematodes (EPNs) form specific mutualistic associations with bioluminescent enterobacteria. In Heterorhabditidis indica, Ochrobactrum spp. was identified beside the symbiont Photorhabdus luminescens but its involvement in the symbiotic association in the EPNs remains unclear. This study describe the population structure and the diversity in Ochrobactrum natural populations isolated from EPNs in the Caribbean basin in order to question the existence of EPN-specialized clones and to gain a better insight into Ochrobactrum-EPNs relationships. EPN-associated Ochrobactrum and Photorhabdus strains were characterized by multi-locus sequence typing, Pulsed-Field Gel Electrophoresis fingerprinting and phenotypic traits. Population study showed the absence of EPN-specialized clones in O. intermedium and O. anthropi but suggested the success of some particular lineages. A low level of genetic and genomic diversification of Ochrobactrum isolated from the natural population of Caribbean nematodes was observed comparatively to the diversity of human-associated Ochrobactrum strains. Correspondences between Ochrobactrum and P. luminescens PFGE clusters have been observed, particularly in the case of nematodes from Dominican Republic and Puerto Rico. O. intermedium and O. anthropi associated to EPNs formed less biofilm than human-associated strains. These results evoke interactions between Ochrobactrum and the EPN symbiotic system rather than transient contamination. The main hypothesis to investigate is a toxic/antitoxic relationship because of the ability of Ochrobactrum to resist to antimicrobial and toxic compounds produced by Photorhabdus.

Attenuated virulence and genomic reductive evolution in the entomopathogenic bacterial symbiont species, Xenorhabdus poinarii.

Bacteria of the genus Xenorhabdus are symbionts of soil entomopathogenic nematodes of the genus Steinernema. This symbiotic association constitutes an insecticidal complex active against a wide range of insect pests. Unlike other Xenorhabdus species, Xenorhabdus poinarii is avirulent when injected into insects in the absence of its nematode host. We sequenced the genome of the X. poinarii strain G6 and the closely related but virulent X. doucetiae strain FRM16. G6 had a smaller genome (500-700 kb smaller) than virulent Xenorhabdus strains and lacked genes encoding potential virulence factors (hemolysins, type 5 secretion systems, enzymes involved in the synthesis of secondary metabolites, and toxin-antitoxin systems). The genomes of all the X. poinarii strains analyzed here had a similar small size. We did not observe the accumulation of pseudogenes, insertion sequences or decrease in coding density usually seen as a sign of genomic erosion driven by genetic drift in host-adapted bacteria. Instead, genome reduction of X. poinarii seems to have been mediated by the excision of genomic blocks from the flexible genome, as reported for the genomes of attenuated free pathogenic bacteria and some facultative mutualistic bacteria growing exclusively within hosts. This evolutionary pathway probably reflects the adaptation of X. poinarii to specific host.

Multilocus sequence-based analysis delineates a clonal population of Agrobacterium (Rhizobium) radiobacter (Agrobacterium tumefaciens) of human origin.

The genus Agrobacterium includes plant-associated bacteria and opportunistic human pathogens. Taxonomy and nomenclature within the genus remain controversial. In particular, isolates of human origin were all affiliated with the species Agrobacterium (Rhizobium) radiobacter, while phytopathogenic strains were designated under the synonym denomination Agrobacterium tumefaciens. In order to study the relative distribution of Agrobacterium strains according to their origins, we performed a multilocus sequence-based analysis (MLSA) on a large collection of 89 clinical and environmental strains from various origins. We proposed an MLSA scheme based on the partial sequence of 7 housekeeping genes (atpD, zwf, trpE, groEL, dnaK, glnA, and rpoB) present on the circular chromosome of A. tumefaciens C58. Multilocus phylogeny revealed that 88% of the clinical strains belong to genovar A7, which formed a homogeneous population with linkage disequilibrium, suggesting a low rate of recombination. Comparison of genomic fingerprints obtained by pulsed-field gel electrophoresis (PFGE) showed that the strains of genovar A7 were epidemiologically unrelated. We present genetic evidence that genovar A7 may constitute a human-associated population distinct from the environmental population. Also, phenotypic characteristics, such as culture at 42°C, agree with this statement. This human-associated population might represent a potential novel species in the genus Agrobacterium.

Gluconobacter as well as Asaia species, newly emerging opportunistic human pathogens among acetic acid bacteria.

Acetic acid bacteria (AAB) are broadly used in industrial food processing. Among them, members of the genera Asaia, Acetobacter, and Granulibacter were recently reported to be human opportunistic pathogens. We isolated AAB from clinical samples from three patients and describe here the clinical and bacteriological features of these cases. We report for the first time (i) the isolation of a Gluconobacter sp. from human clinical samples; (ii) the successive isolation of different AAB, i.e., an Asaia sp. and two unrelated Gluconobacter spp., from a cystic fibrosis patient; and (iii) persistent colonization of the respiratory tract by a Gluconobacter sp. in this patient. We reviewed the main clinical features associated with AAB isolation identified in the 10 documented reports currently available in the literature. Albeit rare, infections as well as colonization with AAB are increasingly reported in patients with underlying chronic diseases and/or indwelling devices. Clinicians as well as medical microbiologists should be aware of these unusual opportunistic pathogens, which are difficult to detect during standard medical microbiological investigations and which are multiresistant to antimicrobial agents. Molecular methods are required for identification of genera of AAB, but the results may remain inconclusive for identification to the species level.

Phylogeny, diversity and host specialization in the phylum Synergistetes with emphasis on strains and clones of human origin.

Members of the phylum Synergistetes have been demonstrated in several environmental ecosystems and mammalian microflorae by culture-independent methods. In the past few years, the clinical relevance of some uncultivated phylotypes has been demonstrated in endodontic infections, and uncultured Synergistetes have been demonstrated in human mouth, gut and skin microbiota. However, Synergistetes are rarely cultured from human samples, and only 17 isolates are currently reported. Twelve members of Synergistetes isolated in the course of various infectious processes, including 3 Jonquetella anthropi, 2 Cloacibacillus evryensis, 2 Pyramidobacter piscolens and 5 unidentified strains, as well as 56 clones obtained by specific PCR from the normal vaginal microflora, were studied. 16S rRNA gene-based phylogeny showed that the clones were grouped into 3 clusters, corresponding to the genus Jonquetella, P. piscolens and one novel Synergistetes taxon. The presence and diversity of Synergistetes were reported for the first time in the vaginal microflora. Synergistetes were found in healthy patients, suggesting that they could play a functional role in human microflorae, but may also act as opportunistic pathogens. Studying the phylogenetic relationships between environmental and mammalian strains and clones revealed clearly delineated independent lineages according to the origin of the sequences.

Negativicoccus succinicivorans gen. nov., sp. nov., isolated from human clinical samples, emended description of the family Veillonellaceae and description of Negativicutes classis nov., Selenomonadales ord. nov. and Acidaminococcaceae fam. nov. in the bacterial phylum Firmicutes.

Three strains of a hitherto unknown, Gram-negative, tiny, anaerobic coccus were collected from human clinical samples originating from skin and soft tissues. The three isolates displayed at least 99.9 % identity in their 16S rRNA gene sequences and more than 99.8 % identity in their dnaK gene sequences. The isolates were affiliated to the family Veillonellaceae, the coccobacillus Dialister micraerophilus being the most closely related species, but there was no more than 91.1 % identity in the 16S rRNA gene sequence between this species and the three isolates. Phylogeny based on the 16S rRNA gene confirmed that the three strains represent a novel and robust lineage within the current family Veillonellaceae. A similar genomic structure was demonstrated for the three isolates by PFGE-based analysis. Morphology and metabolic end products, as well as genotypic and phylogenetic data supported the proposal of the novel genus Negativicoccus gen. nov., with the novel species Negativicoccus succinicivorans sp. nov. [type strain ADV 07/08/06-B-1388(T) (=AIP 149.07(T)=CIP 109806(T)=DSM 21255(T)=CCUG 56017(T)) as type species]. Phylogenetic analyses based on the 16S rRNA gene sequences of members of the phylum Firmicutes and other phyla indicated that the family Veillonellaceae forms a robust lineage clearly separated from those of the classes 'Bacilli', 'Clostridia', Thermolithobacteria and 'Erysipelotrichi' in the phylum Firmicutes. Therefore, we propose that this family is a class-level taxon in the phylum Firmicutes, for which the name Negativicutes classis nov. is proposed, based on the Gram-negative type of cell wall of its members, with the type order Selenomonadales ord. nov. In this order, a novel family, Acidaminococcaceae fam. nov., is proposed and description of the family Veillonellaceae is emended.

Multilocus sequence typing supports the hypothesis that Ochrobactrum anthropi displays a human-associated subpopulation.

BACKGROUND: Ochrobactrum anthropi is a versatile bacterial species with strains living in very diverse habitats. It is increasingly recognized as opportunistic pathogen in hospitalized patients. The population biology of the species particularly with regard to the characteristics of the human isolates is being investigated. To address this issue, we proposed a polyphasic approach consisting in Multi-Locus Sequence Typing (MLST), multi-locus phylogeny, genomic-based fingerprinting by pulsed-field gel electrophoresis (PFGE) and antibiotyping. RESULTS: We tested a population of 70 O. anthropi clinical (n = 43) and environmental (n = 24) isolates as well as the type strain O. anthropi ATCC49188T and 2 strains of Ochrobactrum lupini and Ochrobactrum cytisi isolated from plant nodules. A Multi-Locus Sequence Typing (MLST) scheme for O. anthropi is proposed here for the first time. It was based on 7 genes (3490 nucleotides) evolving mostly by neutral mutations. The MLST approach suggested an epidemic population structure. A major clonal complex corresponded to a human-associated lineage since it exclusively contained clinical isolates. Genomic fingerprinting separated isolates displaying the same sequence type but it did not detect a population structure that could be related to the origin of the strains. None of the molecular method allowed the definition of particular lineages associated to the host-bacteria relationship (carriage, colonisation or infection). Antibiotyping was the least discriminative method. CONCLUSION: The results reveal a human-associated subpopulation in our collection of strains. The emergence of this clonal complex was probably not driven by the antibiotic selective pressure. Therefore, we hypothesise that the versatile species O. anthropi could be considered as a human-specialized opportunistic pathogen.

Plastic architecture of bacterial genome revealed by comparative genomics of Photorhabdus variants.

BACKGROUND: The phenotypic consequences of large genomic architecture modifications within a clonal bacterial population are rarely evaluated because of the difficulties associated with using molecular approaches in a mixed population. Bacterial variants frequently arise among Photorhabdus luminescens, a nematode-symbiotic and insect-pathogenic bacterium. We therefore studied genome plasticity within Photorhabdus variants. RESULTS: We used a combination of macrorestriction and DNA microarray experiments to perform a comparative genomic study of different P. luminescens TT01 variants. Prolonged culturing of TT01 strain and a genomic variant, collected from the laboratory-maintained symbiotic nematode, generated bacterial lineages composed of primary and secondary phenotypic variants and colonial variants. The primary phenotypic variants exhibit several characteristics that are absent from the secondary forms. We identify substantial plasticity of the genome architecture of some variants, mediated mainly by deletions in the 'flexible' gene pool of the TT01 reference genome and also by genomic amplification. We show that the primary or secondary phenotypic variant status is independent from global genomic architecture and that the bacterial lineages are genomic lineages. We focused on two unusual genomic changes: a deletion at a new recombination hotspot composed of long approximate repeats; and a 275 kilobase single block duplication belonging to a new class of genomic duplications. CONCLUSION: Our findings demonstrate that major genomic variations occur in Photorhabdus clonal populations. The phenotypic consequences of these genomic changes are cryptic. This study provides insight into the field of bacterial genome architecture and further elucidates the role played by clonal genomic variation in bacterial genome evolution.

Jonquetella anthropi gen. nov., sp. nov., the first member of the candidate phylum 'Synergistetes' isolated from man.

Six clinical isolates of a hitherto unknown, strictly anaerobic, Gram-negative rod showing fastidious growth were subjected to a polyphasic taxonomic study, including phenotypic, genomic and phylogenetic feature analyses. 16S rRNA gene sequenced-based phylogeny revealed that the novel strains represent a homogeneous group distant from any recognized species in the candidate phylum 'Synergistetes'. The novel isolates were most closely related to species of the genus Dethiosulfovibrio, with 88.2-88.7 % 16S rRNA gene sequence similarity. Large-scale chromosome structure and DNA G+C content also differentiated the novel strains from members of the genus Dethiosulfovibrio. The novel strains were asaccharolytic. Major metabolic end products in trypticase/glucose/yeast extract broth were acetic, lactic, succinic and isovaleric acids and the major cellular fatty acids iso-C(15 : 0) and C(16 : 0). Based on the data presented here, a new genus, Jonquetella gen. nov., is proposed with one novel species, Jonquetella anthropi sp. nov. J. anthropi is the first characterized species of the candidate phylum 'Synergistetes' that includes human isolates. The G+C content of the DNA of the type strain of J. anthropi ADV 126(T) (=AIP 136.05(T)=CIP 109408(T)=CCUG 53819(T)) is 59.4 mol%.

Acidaminococcus intestini sp. nov., isolated from human clinical samples.

Eleven strains of a hitherto unknown, Gram-negative, anaerobic coccus were recovered from various human clinical samples of patients hospitalized in two geographically distant French hospitals. These strains displayed the morphology and growth characteristics of those related to the genus Acidaminococcus. The clinical isolates shared at least 99.9 and 99.7 % of their nucleotide positions in the 16S and 23S rRNA gene sequences, respectively. They displayed 95.6 and 88.9 % 16S and 23S rRNA gene sequence similarities, respectively, with Acidaminococcus fermentans. The 16S rRNA-based phylogeny revealed that all the clinical isolates grouped in a statistically well supported cluster separate from A. fermentans. Enzymic activity profiles as well as metabolic end product patterns, including propionic acid production, differentiated the novel bacteria from A. fermentans. Finally, phenotypic, genotypic and phylogenetic data, including large-scale chromosome structure and DNA G+C content, supported the proposal of a novel species of the genus Acidaminococcus, for which the name Acidaminococcus intestini sp. nov. is proposed. The type strain is ADV 255.99(T) (=AIP 283.01(T)=CIP 108586(T)=CCUG 50930(T)).

Ochrobactrum pseudintermedium sp. nov., a novel member of the family Brucellaceae, isolated from human clinical samples.

Three novel Gram-negative, non-fermenting aerobic bacilli were isolated from human clinical samples. They shared more than 99.8 % of the 16S rRNA gene nucleotide positions. The strains were related to Ochrobactrum intermedium with about 97.48 % 16S rRNA gene sequence similarity. In 16S rRNA gene-, dnaK- and rpoB-based phylogenies, the strains were grouped in a lineage that was distinct from other Ochrobactrum species in the family Brucellaceae. Fatty acid composition, polar lipids, quinone system, DNA-DNA relatedness, genome organization, and physiological and biochemical data differentiated these isolates from recognized species of the genus Ochrobactrum. The three clinical strains therefore represent a novel species within the genus Ochrobactrum, for which the name Ochrobactrum pseudintermedium sp. nov., is proposed. The type strain is ADV31(T) (=CIP 109116(T)=DSM 17490(T)). The DNA G+C content of strain ADV31(T) was 54.5 mol%.

Dialister micraerophilus sp. nov. and Dialister propionicifaciens sp. nov., isolated from human clinical samples.

Seventeen anaerobic, Gram-negative, tiny coccobacilli were collected in France from various human clinical samples. Biochemical analyses as well as molecular studies, including 16S rRNA and dnaK gene sequencing, affiliated all the isolates to the genus Dialister. However, 16S rRNA and dnaK gene sequence similarities were below 95.2 and 79.7 %, respectively, when comparisons were performed with the currently described species Dialister pneumosintes and Dialister invisus. Two clusters consisting of 13 and four isolates could be differentiated. 16S rRNA- and dnaK-based phylogeny confirmed that these two clusters represent two novel and distinct lineages within the genus Dialister. Finally, phenotypic, genotypic and phylogenetic data supported the proposal of the two novel species Dialister micraerophilus sp. nov. (type strain ADV 04.01T=AIP 25.04T=CIP 108278T=CCUG 48837T) and Dialister propionicifaciens sp. nov. (type strain ADV 1053.03T=AIP 26.04T=CIP 108336T=CCUG 49291T). The G+C content of the DNA of the D. micraerophilus type strain is 36.3 mol%. On the basis of 16S rRNA gene sequence analysis, 11 isolates originating from Canada could also be affiliated to D. micraerophilus sp. nov., and were included in the species description.

Molecular and phenotypic features for identification of the opportunistic pathogens Ochrobactrum spp.

Among the six species characterized within the genus Ochrobactrum, Ochrobactrum anthropi and Ochrobactrum intermedium are currently reported as opportunistic pathogens in humans. Since the species identification is mainly based on 16S rDNA analysis, the aim of this study was to search for other characteristics useful for Ochrobactrum species discrimination. Ribotyping, morphological and biochemical analyses, and antimicrobial susceptibility testing were performed for a panel of 35 clinical isolates, first identified to the species level using 16S rDNA sequencing. Type and reference strains of five Ochrobactrum species were comparatively analysed. Commercial identification systems such as API 20NE and VITEK 2 were tested for their ability to identify Ochrobactrum anthropi and to detect other members of the genus Ochrobactrum. An improved protocol for the identification of Ochrobactrum spp. by routine medical microbiology practices is proposed: isolation of a non-fastidious non-fermenting oxidase-positive Gram-negative rod resistant to all beta-lactams except imipenem indicates the genus Ochrobactrum, and the API 20NE system confirms the genus identification for most strains, whereas the VITEK 2 system using ID-GNB cards was less powerful. Urease activity, the mucoidy of the colonies, growth at 45 degrees C on tryptic soy agar, and susceptibility to colistin, tobramycin and netilmicin should be considered as differential characteristics for identification of O. anthropi and O. intermedium to the species level. However, definitive identification depends on genotyping methods.

Pulsed-field gel electrophoresis to study the diversity of whole-genome organization in the genus Ochrobactrum.

The alpha-proteobacterial genus Ochrobactrum groups together organisms that display varied life-styles, such as free-living bacteria, members of rhizosphere and soil, nitrogen-fixing bacteria in plant nodules, xenobiotic-degrading bacteria, colonizers of nematodes and insects, and opportunistic human pathogens. The genomes of nine strains of Ochrobactrum anthropi and eight strains of Ochrobactrum intermedium were analyzed by pulsed-field gel electrophoresis of the whole genome and of I-CeuI digestion fragments. All isolates and type strains of O. anthropi and O. intermedium possessed two high-molecular-weight circular replicons identified as two independent chromosomes on the basis of 16S rDNA hybridization. The genome of the type strain of Ochrobactrum tritici, Ochrobactrum grignonense, and Ochrobactrum gallinifaecis also contained two circular chromosomes. The megaplasmid content was highly variable even among strains in the same species, leading to whole-genome sizes that ranged from 5.060 to 8.300 Mbp and from 4.690 to 7.680 Mbp for O. anthropi and O. intermedium, respectively. This exceptional level of genomic diversity could be related to the adaptability of Ochrobactrum spp. to various ecological niches.

Molecular identification of the first human isolate belonging to the Veillonella ratti-Veillonella criceti group based on 16S rDNA and dnaK gene sequencing.

Anaerobic gram-negative cocci belonging to the genus Veillonella are currently subdivided into eight species. Among them, four have been isolated either from human flora or from clinical samples during infectious processes: Veillonella atypica, V. dispar, V. parvula and V. montpellierensis. To date, the four other species have only been found in animals. In particular, V. ratti and V. criceti, since their characterization, have been exclusively reported from mouth and intestine of rodents. We report here for the first time a human isolate belonging to the V. ratti-V. criceti group recovered in mixed aerobic-anaerobic flora from a semen sample in a 24-year-old man attending the urology unit of our hospital for infertility. Identification of the isolate was based on 16S ribosomal DNA and dnaK gene sequencing. Since then, two strains of Veillonella sp. recovered from semen samples remained unidentified to the species level, and 16S rDNA-based phylogenetic analysis revealed that they might represent novel taxa within the genus Veillonella. Taken together, these observations suggest that host restriction may not exist in the genus Veillonella and that bacterial diversity remains underestimated both within this genus and in human semen.

Veillonella montpellierensis sp. nov., a novel, anaerobic, Gram-negative coccus isolated from human clinical samples.

Three strains of a hitherto unknown, Gram-negative, anaerobic coccus were isolated from human samples. At the phenotypic level, the isolates displayed all the characteristics of bacteria belonging to the genus Veillonella. Sequence analysis revealed that the three strains shared >99.5% similarity in 16S rDNA sequence and >98.4% similarity in dnaK sequence. The three unknown strains formed a separate subclade that was clearly remote from Veillonella species of human and animal origin. Based on these results, the three strains were considered to represent a novel species within the genus Veillonella, for which the name Veillonella montpellierensis is proposed. The type strain of the species is ADV 281.99T (=CIP 107992T=CCUG 48299T).