Bosea rubneri sp. nov. Isolated from Organically Grown Allium cepa.

Strain ZW T0_25T was isolated from an onion sample (Allium cepa var. Hytech F1) within a storage trial and proofed to be a novel, aerobic, Gram-stain negative, rod-shaped bacterial strain. Analyses of the 16S rRNA gene sequence and of the whole draft genome sequences, i.e., digital DNA-DNA hybridization (dDDH), Average Nucleotide Identity (ANI) and Average Amino Acid Identity (AAI) showed that this strain represents a new species of the genus Bosea. The genome size of strain ZW T0_25T is 6.19 Mbp, and the GC content is 66.9%. As whole cell sugars, rhamnose, ribose and glucose were identified. Ubiquinone Q-10 is the major respiratory quinone with 97.8%. Polar lipids in strain ZW T0_25T are composed of one phosphatidylethanolamine, one phosphatidylglycerol, one aminophospholipid, two aminolipids, one glycolipid and two phospholipids whereas the fatty acid profile predominantly consists of C18:1 w7c (63.3%), C16:1 w7c (19.5%) and C16:0 (7.1%). Phenotypic traits were tested in the wet lab as well as predicted in silico from genome data. Therefore, according to this polyphasic approach, the new name Bosea rubneri sp. nov. with the type strain ZW T0_25T (= DSM 116094 T = LMG 33093 T) is proposed.

Rathayibacter rubneri sp. nov. isolated from Allium cepa var. Rijnsburger, an onion landrace.

The novel, aerobic, Gram-stain-positive, rod-shaped bacterial strain, ZW T2_19T, was isolated from an onion sample (Allium cepa var. Rijnsburger). Analyses of the 16S rRNA gene sequence revealed that ZW T2_19T represented a member of the genus Rathayibacter but may represent a novel species of this genus. Analyses of the whole draft genome sequences, i.e. digital DNA-DNA hybridisation (dDDH) and average nucleotide identity (ANI) of ZW T2_19T and all type strains of species of the genus Rathayibacter confirmed that ZW T2_19T represents a novel species of the genus Rathayibacter. The genome size of ZW T2_19T is 4.01 Mbp and the DNA G+C content is 71.8 mol%. Glucose, mannose, rhamnose and ribose were detected as whole-cell sugars of ZW T2_19T. The major respiratory quinone of ZW T2_19T is menaquinone MK-10, at 78.9 %. The detected peptidoglycan type in ZW T2_19T is a variant of type B2γ with {Gly} [l-diaminobutyric acid (l-DAB)/l-homoserine (l-Hse)] d-Glu-l-DAB. Polar lipids in ZW T2_19T consisted of one diphosphatidylglycerol, one phosphatidylglycerol, seven glycolipids, one phospholipid and one lipid. The fatty acid profile of ZW T2_19T predominantly consisted of anteiso-C15 : 0 (53 %), iso-C16 : 0 (21 %) and anteiso-C17 : 0 (18 %). In addition, API 20NE, API 50CH, API Coryne, API ZYM, antibiotic susceptibility, haemolysis and growth at different temperatures and with different supplements was investigated. On the basis of the results obtained using this polyphasic approach, including molecular, phenotypic and biochemical analyses, we propose the novel species Rathayibacter rubneri with the type and only strain ZW T2_19T (= DSM 114294T = LMG 32700T).

Metabolism of glyphosate by the human fecal microbiota.

Glyphosate is the most frequently used herbicide worldwide and its application is under discussion due to health concerns. As humans may be exposed to glyphosate, the present study investigated the metabolism of glyphosate by the human fecal microbiota in vitro. Human fecal samples were collected from 15 different volunteers and fecal suspensions were prepared. The human fecal suspension samples were incubated with glyphosate under strictly anaerobic conditions and glyphosate degradation was investigated. Neither a degradation of glyphosate, nor a formation of AMPA (aminomethylphosphonic acid), the known microbial metabolite in soil, was detected. In conclusion, the microbiota of human fecal suspensions did not metabolize glyphosate under the conditions used in our study which hints at the assumption that transformation of glyphosate by the gut microbiota seems to be negligible in humans.

Adlercreutzia rubneri sp. nov., a resveratrol-metabolizing bacterium isolated from human faeces and emended description of the genus Adlercreutzia.

The novel, anaerobic, Gram-positive, rod-shaped bacterial strain, ResAG-91T, was isolated from a faecal sample of a male human volunteer. Analysis of the 16S rRNA gene sequence revealed that strain ResAG-91T showed high similarity to the type strains of Adlercreutzia equolifaciens subsp. equolifaciens and Adlercreutzia equolifaciens subsp. celatus. Analysis of the whole draft genome sequences, i.e. digital DNA-DNA hybridization (dDDH) and average nucleotide identity (ANI), of strain ResAG-91T and the type strains of Adlercreutzia species revealed that strain ResAG-91T represents a novel species of the genus Adlercreutzia. The genome size of strain ResAG-91T is 2.8 Mbp and the G+C content is 63.3 mol%. The major respiratory quinone of strain ResAG-91T was MMK-5 (methylmenaquinone). Major cellular fatty acids were C15 : 0 anteiso, C14 : 0 iso and C14 : 0 2-OH. Galactose and ribose were detected as major whole cell sugars. Furthermore, the peptidoglycan type of strain ResAG-91T was A1γ with meso-diaminopimelic acid. The polar lipids were phosphatidylglycerol, diphosphatidylglycerol, one unidentified lipid, three unidentified phospholipids and five unidentified glycolipids. Strain ResAG-91T was able to metabolize the stilbene resveratrol into dihydroresveratrol. On the basis of this polyphasic approach, including phenotypical, molecular (16S rRNA gene and whole genome sequencing) and biochemical (fatty acids, quinones, polar lipids, peptidoglycan, whole cell sugars, Rapid ID32A and API20A) analyses, we propose the novel species Adlercreutzia rubneri sp. nov. with the type and only strain ResAG-91T (=DSM 111416T=JCM 34176T=LMG 31897T).

Genome analysis reveals that the correct name of type strain Adlercreutzia caecicola DSM 22242T is Parvibacter caecicola Clavel et al. 2013.

The strain Adlercreutzia caecicola DSM 22242T (=CCUG 57646T=NR06T) was taxonomically described in 2013 and named as Parvibacter caecicola Clavel et al. 2013. In 2018, the name of the strain DSM 22242T was changed to Adlercreutzia caecicola (Clavel et al. 2013) Nouioui et al. 2018 due to taxonomic investigations of the closely related genera Adlercreutzia, Asaccharobacter and Enterorhabdus within the phylum Actinobacteria. However, the first whole draft genome of strain DSM 22242T was published by our group in 2019. Therefore, the genome was not available within the study of Nouioui et al. (2018). The results of the polyphasic approach within this study, including phenotypic and biochemical analyses and genome-based taxonomic investigations [genome-wide average nucleotide identity (gANI), alignment fraction (AF), average amino acid identity (AAI), percentage of orthologous conserved proteins (POCP) and genome blast distance phylogeny (GBDP) tree], indicated that the proposed change of the name Parvibacter caecicola to Adlercreutzia caecicola was not correct. Therefore, it is proposed that the correct name of Adlercreutzia caecicola (Clavel et al. 2013) Nouioui et al. 2018 strain DSM 22242T is Parvibacter caecicola Clavel et al. 2013.

Fermentation of African nightshade leaves with lactic acid bacterial starter cultures.

The interest in the consumption of African indigenous leafy vegetables increased in African countries, e.g. Kenya, within the last years. One example of African indigenous leafy vegetables is African nightshade (Solanum scabrum) which is nutritious, rich in proteins and micronutrients and therefore could contribute to a healthy diet. African nightshade has several agricultural advantages. However, the most important disadvantage is the fast perishability which leads to enormous post-harvest losses. In this study, we investigated the fermentation of African nightshade as a post-harvest processing method to reduce post-harvest losses. The two lactic acid bacterial starter strains Lactiplantibacillus plantarum BFE 5092 and Limosilactobacillus fermentum BFE 6620 were used to inoculate fermentations of African nightshade leaves with initial counts of 106-107 cfu/ml. Uninoculated controls were conducted for each fermentation trial. Fermentations were performed both in Kenya and in Germany. The success of the inoculated starter cultures was proven by the measurement of pH values and determination of lactic acid concentration. Lactobacilli strains dominated the microbiota of the starter inoculated samples in contrast to the non-inoculated controls. This was supported by classical culture-dependent plating on different microbiological media as well as by the culture-independent molecular biological methods denaturing gradient gel electrophoresis and 16S rRNA gene high-throughput amplicon sequencing. We could demonstrate that the use of the selected starter cultures for fermentation of African nightshade leaves led to controlled and reliable fermentations with quick acidification. Thus, controlled fermentation with appropriate starter cultures is a promising method for post-harvest treatment of African nightshade leaves.

Influence of salt concentration and iodized table salt on the microbiota of fermented cucumbers.

The fermentation of vegetables is a traditional preservation method, that experiences a renaissance even in domestic households. Table salt is added to the fermentation batches to favor the growth of lactic acid bacteria usually. On an industrial scale, the fermentation brine is typically prepared with non-iodized table salt. In our study, we investigated the microbiota of cucumber fermentations using culture-dependent and -independent methods. We could show that the fermentation process of cucumbers and the involved microbiota is influenced by the concentration of table salt and not by the use of iodized table salt. Therefore, we conclude that the use of iodized table salt does not negatively affect the fermentation process. We could verify that iodine permeates the cucumbers by diffusion, leading to satisfactory iodine concentrations in the final food product. The industrial use of iodized table salt in food fermentations could contribute to maintain a constant iodine supply to the general public.

Draft Genome Sequences of 13 Isolates of Adlercreutzia equolifaciens, Eggerthella lenta, and Gordonibacter urolithinfaciens, Isolated from Human Fecal Samples in Karlsruhe, Germany.

Here, we report the annotated draft genome sequences of 13 Eggerthellaceae strains isolated from fecal samples from two healthy human volunteers in Karlsruhe, Germany, i.e., Adlercreutzia equolifaciens ResAG-91, Eggerthella lenta MRI-F 36, MRI-F 37, MRI-F 40, ResAG-49, ResAG-88, ResAG-121, and ResAG-145, and Gordonibacter urolithinfaciens ResAG-5, ResAG-26, ResAG-43, ResAG-50, and ResAG-59.

Draft Genome Sequences of Type Strains of Adlercreutzia muris and Ellagibacter urolithinifaciens, Belonging to the Family Eggerthellaceae.

Here, we report the annotated draft genome sequences of two type strains belonging to the family Eggerthellaceae within the class Coriobacteriia (phylum Actinobacteria), Adlercreutzia muris WCA-131-CoC-2 (= DSM 29508 = KCTC 15543) and Ellagibacter urolithinifaciens CEBAS 4A (= CCUG 70284 = DSM 104140).

Antibiotics resistance and toxin profiles of Bacillus cereus-group isolates from fresh vegetables from German retail markets.

BACKGROUND: This study aimed to evaluate the safety of raw vegetable products present on the German market regarding toxin-producing Bacillus cereus sensu lato (s.l.) group bacteria. RESULTS: A total of 147 B. cereus s.l. group strains isolated from cucumbers, carrots, herbs, salad leaves and ready-to-eat mixed salad leaves were analyzed. Their toxinogenic potential was assessed by multiplex PCR targeting the hemolysin BL (hbl) component D (hblD), non-hemolytic enterotoxin (nhe) component A (nheA), cytotoxin K-2 (cytK-2) and the cereulide (ces) toxin genes. In addition, a serological test was used to detect Hbl and Nhe toxins. On the basis of PCR and serological results, none of the strains were positive for the cereulide protein/genes, while 91.2, 83.0 and 37.4% were positive for the Hbl, Nhe and CytK toxins or their genes, respectively. Numerous strains produced multiple toxins. Generally, strains showed resistance against the ß-lactam antibiotics such as penicillin G and cefotaxim (100%), as well as amoxicillin/clavulanic acid combination and ampicillin (99.3%). Most strains were susceptible to ciprofloxacin (99.3%), chloramphenicol (98.6%), amikacin (98.0%), imipenem (93.9%), erythromycin (91.8%), gentamicin (88.4%), tetracycline (76.2%) and trimethoprim/sulfamethoxazole combination (52.4%). The genomes of eight selected strains were sequenced. The toxin gene profiles detected by PCR and serological test mostly agreed with those from whole-genome sequence data. CONCLUSIONS: Our study showed that B. cereus s.l. strains encoding toxin genes occur in products sold on the German market and that these may pose a health risk to the consumer if present at elevated levels. Furthermore, a small percentage of these strains harbor antibiotic resistance genes. The presence of these bacteria in fresh produce should, therefore, be monitored to guarantee their safety.

Gordonibacter faecihominis is a later heterotypic synonym of Gordonibacter urolithinfaciens.

In this study, the phylogenetic position of Gordonibacter faecihominis and Gordonibacter urolithinfaciens was investigated using phenotypic and molecular (rep-PCR, ARDRA, 16S rRNA gene sequencing and whole-genome sequencing) methods. Our results show that Gordonibacter faecihominis cannot be distinguished from Gordonibacter urolithinfaciens on the basis of the results of this polyphasic approach. Therefore, it is proposed that the two species Gordonibacter faecihominis and Gordonibacter urolithinfaciens belong to the same species.

Draft Genome Sequences of Type Strains of Gordonibacter faecihominis, Paraeggerthella hongkongensis, Parvibacter caecicola,Slackia equolifaciens, Slackia faecicanis, and Slackia isoflavoniconvertens.

Here, we report the annotated draft genome sequences of six type strains of the family Eggerthellaceae, Gordonibacter faecihominis JCM 16058, Paraeggerthella hongkongensis DSM 16106, Parvibacter caecicola DSM 22242, Slackia equolifaciens DSM 24851, Slackia faecicanis DSM 17537, and Slackia isoflavoniconvertens DSM 22006.

Draft Genome Sequence of Salmonella enterica subsp. enterica Serovar Enteritidis MS 501, a Potential Human Pathogen Isolated from Red Lettuce (Lactuca sativa var. capitata) in Karlsruhe, Germany.

We report here the genome sequence of Salmonella enterica subsp. enterica serovar Enteritidis MS 501, a potential human pathogen isolated from red lettuce (Lactuca sativa var. capitata) in Karlsruhe, Germany. The assembled genome size was 4,700,322 bp. A total of 4,560 coding genes, 16 rRNAs, 78 tRNAs, and 15 noncoding RNAs were predicted.

Rubneribacter badeniensis gen. nov., sp. nov. and Enteroscipio rubneri gen. nov., sp. nov., new members of the Eggerthellaceae isolated from human faeces.

Two novel, anaerobic, Gram-positive, rod-shaped bacterial strains, ResAG-85T and ResAG-96T, were isolated from a faecal sample of a male human. 16S rRNA gene sequences analyses indicated that these strains represent a distinct lineage within the family Eggerthellaceae. Strain ResAG-85T showed 92.3 % similarity to the type strains of the genera Eggerthella and Gordonibacter. Strain ResAG-96T clustered together with Paraeggerthella hongkongensis and the newly (but not validly) published genus 'Arabia massiliensis' (94.8 % similarity). Analysis of quinones revealed that MK-5 (21 % in ResAG-85T and 95 % in ResAG-96T) and MK-7 (53 % in strain ResAG-85T) were present, which were described for the first time for members of the Eggerthellaceae. Furthermore, MK-6 was present in both strains (25 % ResAG-85T and 5 % in ResAG-96T). The polar lipids detected in ResAG-85T and ResAG-96T consisted of eight and six glycolipids, respectively. Both strains possessed three phospholipids, one phosphatidylglycerol and one diphosphatidylglycerol. Analysis of fatty acids revealed that the percentage of total branched fatty acids was relatively high in comparison to related strains with 42 and 50 % of strains ResAG-85T and ResAG-96T but comparable to the value obtained for Gordonibacter pamelaeae DSM 19378T. On the basis of this polyphasic approach including molecular (16S rRNA gene sequencing) and biochemical methods (analysis of fatty acids, quinones, polar lipids, Rapid ID 32A and API 20A), the new genera and species Rubneribacter badeniensis with ResAG-85T (=DSM 105129T=JCM 32272T) and Enteroscipio rubneri with ResAG-96T (=DSM 105130T=JCM 32273T) as the type and only strains are described.

Influence of Different Nanomaterials on Growth and Mycotoxin Production of Penicillium verrucosum.

Nanoparticles are ubiquitous in the environment. They originate from anthropogenic or natural sources or they are intentionally produced for different purposes. There exist manifold applications of nanoparticles in modern life leading unavoidably to a confrontation and interaction between nanomaterial and living organisms. Based on their wide distribution tending to increase steadily, the influence of particles based on silica and silver, exhibiting nominal sizes between 0.65 nm and 200 nm, on the physiology of the mycotoxigenic filamentous fungus Penicillium verrucosum was analyzed. The applied concentration and time-point, the size and the chemical composition of the particles was shown to have a strong influence on growth and mycotoxin biosynthesis. On microscopic scale it could be shown that silver nanoparticles attach to the mycelial surface. Moreover, silver nanoparticles with 0.65 nm and 5 nm in size were shown to internalize within the cell, form agglomerates in the cytoplasm and associate to cell organelles.

Comparative proteome analysis of Penicillium verrucosum grown under light of short wavelength shows an induction of stress-related proteins associated with modified mycotoxin biosynthesis.

In this study the differentially expressed protein population of Penicillium verrucosum grown either in the dark or under light with a wavelength of 450nm has been analyzed. Light of short wavelength led to oxidative stress in the fungal cell; under this condition the mycotoxin biosynthesis revealed a mutual shift from ochratoxin A to citrinin. Using a proteomic approach combining an optimized protein extraction method with 2-dimensional SDS-PAGE followed by HPLC-ESI-TOF-MS/MS mass spectrometric analysis, initially 56 significantly differential proteins (light vs. dark) were detected comprising proteins of a broad range of isoelectric points and molecular masses. In total, 46 proteins could be identified further by database query, most of these proteins are assumed to be involved in response to stress (e.g. antioxidative proteins, heat shock proteins) and general metabolic processes (e.g. glycolysis, ATP supply). Proteome analyses are necessary to unravel the regulation of secondary metabolite biosynthesis at a translational level. This may enable identification of proteins which are involved in mycotoxin biosynthesis, adaption processes or even stress compensation mechanisms. This study depicts the first proteome analysis of P. verrucosum.

Intraspecific variability of HOG1 phosphorylation in Penicillium verrucosum reflects different adaptation levels to salt rich habitats.

Mycotoxin biosynthesis in Penicillium verrucosum is modulated by different molecular regulation mechanisms. One important mechanism is the HOG1 (high osmolarity glycerol) MAP kinase signaling pathway. In a comparative analysis three different P. verrucosum strains were selected from six strains with different ability to produce the mycotoxins ochratoxin and citrinin. The fungal strains were grown on laboratory medium supplemented with different concentrations of NaCl. It could be shown that there exists an interrelationship between the growth rate, the level of HOG phosphorylation and the mycotoxin biosynthesis under the respective growth condition. The weak to non ochratoxin producing P. verrucosum strain, BFE875, showed only a poor growth rate but the strongest HOG1 phosphorylation; the strong ochratoxin and citrinin producing strain BFE575 showed a reasonable HOG1 phosphorylation with an average growth rate; and the strong ochratoxin and weak citrinin producing strain BFE495 showed only a poor phosphorylation but the highest growth rate in 7days of incubation at 25°C. The magnitude of phosphorylation of the HOG1 protein seems to be inversely correlated with the degree of adaption of the fungus to hyperosmotic growth conditions.