Beer spoilage causes huge economic losses for the brewing industry. Only a few obligate anaerobic bacteria are capable of growing in beer. They can spoil the batches after brewing and during maturation, thereby causing especially serious damage. Pectinatus frisingensis is one of the most dreaded species of anaerobic beer spoilers. It causes turbidity and acidity as well as off flavors. While the growth-limiting stressors of beer have been well studied, the underlying mechanisms of adaptation are still largely unknown.A transcriptional study was conducted on P. frisingensis TMW 2.2471 by exposing it to ethanol stress of 4.76% (v/v) and acid stress at pH 4.04 separately. Both experiments were executed in alcohol free beer (AFB) and peptone yeast extract glucose medium (PYG). The transcriptomic responses of P. frisingensis strain TMW 2.2471 to ethanol and acid stress showed several differently upregulated genes. P. frisingensis reacted to both stressors by upregulating heat shock protein 20 and UvrABC endonuclease for repair processes. Ethanol stress additionally caused the upregulation of co-chaperone GroES and Clp protease, while expressing acetolactate synthase and decarboxylase only under acid stress. Our findings suggest that P. frisingensis is capable of a broad response in an unexpectedly large number of metabolic functions. Its survival is based on combining universal and exclusive stress responses to ethanol and acid stress, the two main stressors presented by beer.

Megasphaera sueciensis is an obligate anaerobic, gram-negative beer spoilage bacterium causing unwanted off-flavors and turbidity. Here we report the complete and circular genome of 2.9 Mbp with 2,696 predicted coding sequences of the type strain Megasphaera sueciensis DSM 17042.

Traditional sausage in the Republic of Kosovo has been produced for centuries as a traditional method of preserving the nutritional value of meat. In sausage fermentation, natural microbiota such as lactic acid bacteria (LAB) and Micrococcaceae usually participate; these are not only critical for ensuring product safety and flavor development but also represent significant biotechnological potential. The purpose of this study was to analyze traditional fermented sausage, in terms of production practices and hygiene, throughout the production and storage phases. Samples in three stages of production and maturation were analyzed for microbiota, pH, and water activity level. Our results show that the main changes in the bacterial populations from 0 to 7 days of storage included increases in the total numbers of viable mesophilic aerobic bacteria (LAB) and Micrococcaceae (MC). However, the Enterobacteriaceae and coliforms (EC) count showed a significant decrease (p < 0.05) in 1.60 ± 1.62 lg cfu/g by day 14. In conclusion, the number of EC in the traditional sausage was decreased during storage, while LAB and MC were stable, data that indicate the safety and quality of this product. No differences regarding the production practices and storage of traditional sausage were observed, based on the data from the butchers who participated in this study.

The only two commercially available strains of Megasphaera paucivorans were cultivated and their genomes sequenced. As recurring beer spoiling bacteria, they cause unwanted turbidity and unpleasant odors. Their genomes harbor a number of putative defense mechanisms explaining their much-needed resilience to survive in the brewing environment.

Five strains of Gram-stain-positive, non-motile, and coccoid-shaped bacteria, designated as TMW 2.2395T, TMW 2.2628, TMW 2.2670, TMW 2.2756 and TMW 2.2757 were isolated from feces and nasal mucosa of wild living roe deer (Capreolus capreolus) and red deer (Cervus elaphus), respectively. The isolates share identical 16S rRNA gene sequences and are classified within the genus Macrococcus based on 16S and MALDI-TOF MS analyses. Phylogenetic analysis revealed that average nucleotide identity (ANI) values were below the accepted thresholds for prokaryotic species delineation, with the type strains of M. goetzii, M. bohemicus and M. epidermidis showing the highest relatedness values (79.59 %, 79.30 % and 79.52 %, respectively). Digital DNA-DNA hybridization (dDDH) values were below 23 % confirming that all five strains belong to a new species within the genus Macrococcus. All strains were catalase and oxidase positive, grew optimally at 37 °C and pH 7.0, but demonstrated considerably lower salt tolerance (< 7.5 % w/v nit iterativ) compared to other species in the genus. Moreover, the data of this study suggests that members of this new species possess an intrinsic resistance to fosfomycin, putatively mediated by a fosB homologue. The genomes range from 2.4 and 2.5 Mb in size, with a G+C content of 34.8 to 35.1 %. The primary respiratory quinone of the type strain TMW 2.2395T is Menaquinone 6 (MK-6, 98.6 %), the cell wall peptidoglycan type is A3α L-Lys-Gly3-L-Ser and the major fatty acids are C14:0, C16:0 and C18:0. Phylogenetic, phenotypic and chemotaxonomic data collectively suggest that these strains represent a novel species of the genus Macrococcus, for which the name Macrococcus capreoli sp. nov. is proposed. The type strain is TMW 2.2395T = DSM 113939T = LMG 32618T.

Beer spoiling bacteria represent a major concern for the brewing industry. Anaerobic spoilers of the genus Megasphaera are especially dreadful. Until now, the genome analysis of Megasphaera spp. was limited to mammal-associated species, yet beer borne Megasphaera species and their abilities to tolerate the multiple high stress factors in their niche were broadly overlooked. This study is the first to carry out an in silico genome comparison of ten strains from three different beer spoiling Megasphaera species (M. cerevisiae, M. paucivorans and M. sueciensis). We experimentally show a hitherto unseen resilience of beer spoiling Megasphaera strains, which can grow in lager beer with an isoα-acids content of up to 120 IBU, an ethanol concentration of up to 7.0 % (v/v) and at pH 4.0. The genomes revealed genes appearing to aid survival in the harsh brewing environment mainly comprised of hop, ethanol and acid stress.

Background Autolysis by cellular peptidoglycan hydrolases (PGH) is a well-known phenomenon in bacteria. During food fermentation, autolysis of starter cultures can exert an accelerating effect, as described in many studies on cheese ripening. In contrast, very little is known about autolysis of starter cultures used in other fermentations. Staphylococcus (S.) carnosus is often used in raw sausage fermentations, contributing to nitrate reduction and flavor formation. In this study, we analyzed the influence of PGHs of the strains S. carnosus TMW 2.146 and S. carnosus TMW 2.2525 on their autolytic behavior. The staphylococcal major autolysin (Atl), a bifunctional enzyme with an N-acetylmuramoyl-L-alanine amidase and a glucosaminidase as an active site, is assumed to be the enzyme by which autolysis is mainly mediated. Results AtlC mutant strains showed impaired growth and almost no autolysis compared to their respective wild-type strains. Light microscopy and scanning electron microscopy showed that the mutants could no longer appropriately separate from each other during cell division, resulting in the formation of cell clusters. The surface of the mutants appeared rough with an irregular morphology compared to the smooth cell surfaces of the wild-types. Moreover, zymograms showed that eight lytic bands of S. carnosus , with a molecular mass between 140 and 35 kDa, are processed intermediates of AtlC. It was noticed that additional bands were found that had not been described in detail before and that the banding pattern changes over time. Some bands disappear entirely, while others become stronger or are newly formed. This suggests that AtlC is degraded into smaller fragments over time. A second knockout was generated for the gene encoding a N-acetylmuramoyl-L-alanine amidase domain-containing protein. Still, no phenotypic differences could be detected in this mutant compared to the wild-type, implying that the autolytic activity of S. carnosus is mediated by AtlC. Conclusions In this study, two knockout mutants of S. carnosus were generated. The atlC mutant showed a significantly altered phenotype compared to the wild-type, revealing AtlC as a key factor in staphylococcal autolysis. Furthermore, we show that Atl is degraded into smaller fragments, which are still cell wall lytic active.

AIMS:Temperate phages insert their genome into the host's chromosome. As prophages, they remain latent in the genome until an induction event leads to lytic phage production. When this occurs in a starter culture that has been added to food fermentation, this can impair the fermentation success. This study aimed to analyze prophage inducibility in the Latilactobacillus curvatus TMW 1.591 strain during meat fermentation and investigate whether an induction signal before cryopreservation is maintained during storage and can lead to phage-induced lysis after culture activation. METHODS AND RESULTS:A prophage-free isogenic derivative of the model starter organism, L. curvatus TMW 1.591, was developed as a negative control (L. curvatus TMW 1.2406). Raw meat fermentation was performed with the wild-type (WT) and phage-cured strains. The WT strain produced high numbers of phages (5.2 ± 1.8 × 107 plaque-forming units g-1) in the meat batter. However, the prophage did not significantly affect the meat fermentation process. Induction experiments suggested an acidic environment as a potential trigger for prophage induction. Phage induction by ultraviolet light before strain cryopreservation remains functional for at least 10 weeks of storage. CONCLUSIONS:Intact prophages are active during meat fermentation. However, in this study, this has no measurable consequences for fermentation, suggesting a high resiliency of meat fermentation against phages. Inadequate handling of lysogenic starter strains, even before preservation, can lead to phage introduction into food fermentation and unintended host lysis.

Obligate anaerobic beer spoilage bacteria have been a menace to the brewing industry for several decades. Technological advances in the brewing process aimed at suppressing aerobic spoilers gave rise to problems with obligate anaerobes. In previous studies, the metabolic spectrum of Pectinatus and Megasphaera species has been described, but their metabolism in the beer environment remains largely unknown. We used high-performance anion exchange chromatography with pulsed amperometric detection (HPAEC-PAD) and headspace solid-phase microextraction–gas chromatography–mass spectrometry (HS-SPME-GCMS) to further characterize beer spoiled by 30 different strains from six beer-spoiling species of Pectinatus and Megasphaera (P. cerevisiiphilus, P. frisingensis, P. haikarae, M. cerevisiae, M. paucivorans, and M. sueciensis). We detected differences in carbohydrate utilization and the volatile organic compounds (volatilome) produced during beer spoilage by all six species. We were able to show that glycerol, one of the basic components of beer, is the common carbon source used by all strains. It appears that this carbon source allows for anaerobic beer spoilage by Pectinatus and Megasphaera despite the spoilage-preventing intrinsic barriers of beer (iso-α-acids, ethanol, low pH, scarce nutrients); thus, extrinsic countermeasures are key for prevention.

Lactococcus (Lc.) paracarnosus and the phylogenetically closely related Lc. carnosus species are common members of the microbiota in meat stored under modified atmosphere and at low temperature. The effect of these strains on meat spoilage is controversially discussed. While some strains are known to cause spoilage, others are being studied for their potential to suppress the growth of spoilage and pathogenic bacteria. In this study, Lc. paracarnosus DSM 111017T was selected based on a previous study for its ability to suppress the growth of meat spoilers, including Brochothrix thermosphacta. The mechanism by which this bioprotective strain inhibits competing bacteria and how it contributes to spoilage are not yet known. To answer these two questions, we investigated the effect of four different headspace gas mixtures (simulated air (21 % O2/79 % N2); HiOx-MAP (70 % O2/30 % CO2); nonOx-MAP (70 % N2/ 30 % CO2); simulated vacuum (100 % N2) and the presence of Brochothrix (B.) thermosphacta TMW 2.2101 on the growth and transcriptional response of Lc. paracarnosus DSM 111017T when cultured on a meat simulation agar surface at 4 °C. Analysis of genes specifically upregulated by the gas mixtures used revealed metabolic pathways that may lead to different levels of spoilage metabolites production. We propose that under elevated oxygen levels, Lc. paracarnosus preferentially converts pyruvate from glucose and glycerol to uncharged acetoin/diacetyl instead of lactate to counteract acid stress. Due to the potential production of a buttery off-flavour, the strain may not be suitable as a protective culture in meat packaged under high‑oxygen conditions. 70 % N2/ 30 % CO2, simulated vacuum- and the presence of Lc. paracarnosus inhibited the growth of B. thermosphacta TMW 2.2101. However, B. thermosphacta did not affect gene regulation of metabolic pathways in Lc. paracarnosus, and genes previously predicted to be involved in B. thermosphacta growth suppression were not regulated at the transcriptional level. In conclusion, the study indicates that the gas mixture used in packaging significantly affects the metabolism and spoilage potential of Lc. paracarnosus and its ability to inhibit B. thermosphacta growth.

Restriction modification (RM) systems are known to provide a strong barrier to the exchange of DNA between and within bacterial species. Likewise, DNA methylation is known to have an important function in bacterial epigenetics regulating essential pathways such as DNA replication and the phase variable expression of prokaryotic phenotypes. To date, research on staphylococcal DNA methylation focused mainly on the two species Staphylococcus aureus and S. epidermidis. Less is known about other members of the genus such as S. xylosus, a coagulase-negative commensal of mammalian skin. The species is commonly used as starter organism in food fermentations but is also increasingly considered to have an as yet elusive function in bovine mastitis infections. We analyzed the methylomes of 14 S. xylosus strains using single-molecular, real-time (SMRT) sequencing. Subsequent in silico sequence analysis allowed identification of the RM systems and assignment of the respective enzymes to the discovered modification patterns. Hereby the presence of type I, II, III and IV RM systems in varying numbers and combinations among the different strains was revealed, clearly distinguishing the species from what is known for other members of the genus so far. In addition, the study characterizes a newly discovered type I RM system, encoded by S. xylosus but also by a variety of other staphylococcal species, with a hitherto unknown gene arrangement that involves two specificity units instead of one (hsdRSMS). Expression of different versions of the operon in E. coli showed proper base modification only when genes encoding both hsdS subunits were present. This study provides new insights into the general understanding of the versatility and function of RM systems as well as the distribution and variations in the genus Staphylococcus.

The genus Dellaglioa (D.) actually comprises two species, i.e., D. algida and the recently described species D. carnosa. Both species are adapted to cold and have been typically recovered from meat products. However, their importance has thus far been underestimated, since routine culture-based analysis failed to support their growth. Furthermore, their occurrence on meat packed under high-oxygen MA conditions (HiOx-MAP) is controversial because they have been described as being oxygen-sensitive. In this study, we focused on the targeted isolation of Dellaglioa spp. from HiOx-MAP meat samples and the characterization of our isolates regarding their adaption to HiOx-MAP conditions, their spoilage potential, as well as food safety aspects. We used a medium recently developed specifically for strains of this genus and investigated ten meat batches from seven different suppliers. Our study confirms that the occurrence of Dellaglioa spp. on HiOx-MAP meat is non-sporadic, reaching cell counts ranging from log10 5.8-7.1 CFU/cm2 at a late stage of chilled storage. Autochthonous Dellaglioa spp. and Leuconostoc (L.) gasicomitatum dominated the microbiota of the beef steaks with similar growth behavior. Our results suggest that Dellaglioa spp. benefits from the heme-dependent respiration of oxygen by L. gasicomitatum. Furthermore, whole genome analysis revealed the presence of genes predictively involved in oxidative stress defense, survival, and adaptation in meat environments. Moreover, we predict a weak aminogenic potential of D. algida strains. Tyramine production from tyrosine seems to be a species-specific characteristic of D. carnosa. The extent to which D. algida and D. carnosa occurrence is influenced by or even dependent on the composition of the entire microbiota remains to be investigated.

The study provides a taxonomic characterization of three bacterial strains isolated from high-oxygen modified -atmosphere packaged beef from Germany. The strains of the novel species shared identical 16S rRNA gene sequence to the closely related type strain of Dellaglioa algida. However, the in-silico DNA-DNA hybridization (DDH) values indicate that they belong to a different genomic species. The in silico DDH estimate value between TMW 2.2523T and the type strain of Dellaglioa algida DSM 15638T was only 63.2 %. The whole genome average nucleotide identity blast (ANIb) value of 95.1 % between TMW 2.2523T and the closely related type strain of D. algida was within the recommended threshold value of 95-96 % for bacterial species delineation. Additionally, the phylogenomic analyses based on multi locus sequence alignment (MLSA) showed that strain TMW 2.2523T and additional strains TMW 2.2444 and TMW 2.2533 formed a monophyletic group separate from D. algida strains. Furthermore, tyrosine decarboxylase activity could be attributed to strains of the new proposed species. The results of this polyphasic approach support the affiliation of these strains to a novel species within the genus Dellaglioa for which we propose the name Dellaglioa carnosa sp. nov. The designated respective type strain is TMW 2.2523T (DSM 114968T = LMG 32819T).

Lupine-based seasoning sauce is produced similarly to soy sauces and therefore generates a comparable microbiota and aroma profile. While the koji state is dominated by Aspergillus oryzae, the microbiome of the moromi differs to soy moromi, especially in yeast composition due to the absence of Zygosaccharomyces rouxii and Debaryomyces hansenii as the dominant yeast. In this study, we monitored the addition of a carbohydrate source on the microbiome and aroma profile of the resulting sauce. Compared to previous studies, the usage of a yeast starter culture resulted in a sparsely diverse microbiota that was dominated by D. hansenii and T. halophilus. This led to a pH below 5 even after four months of incubation and most of the measured aroma compounds were pyrazines and acids. The addition of wheat and buckwheat resulted in a temporary change in the yeast consortium with the appearance of Z. rouxii and additional bacterial genera. The aroma profile differs in the presence of pyrazines and esters. Since no significant differences in the taste and odour of wheat-added and buckwheat-added sauce was sensed, both substrates influence the lupine sauce in a similar way.

AIMS Acetic acid bacteria of the genus Bombella have not been reported to produce exopolysaccharides. In this study the formation of fructans by B. apis TMW 2.1884 and B. mellum TMW 2.1889 was investigated. METHODS AND RESULTS Out of eight strains from four different Bombella species, only B. apis TMW 2.1884 and B. mellum TMW 2.1889 showed exopolysaccharide (EPS) formation with 50 g L-1 sucrose as substrate. Both EPS were identified as high molecular weight (HMW) polymers (106-107 Da) by AF4-MALLS/UV and HPSEC-MALLS/RI analyses. Monosaccharide analysis via TFA hydrolysis showed that both EPS are fructans. Determination of glycosidic linkages by methylation analysis revealed mainly 2,6-linked fructofuranose (Fruf) units with additional 1,2-linked Fruf units (10%) and 1,2,6-Fruf branched units (7%). No glycoside hydrolase (GH) 68 family genes that are typically associated with the formation of high molecular weight fructans in bacteria could be identified in the genomes. Through heterologous expression in E. coli Top10, an enzyme of the GH 32 family could be assigned to the catalysis of fructan formation. The identified fructosyltransferases could be clearly differentiated phylogenetically and structurally from other previously described bacterial fructosyltransferases. CONCLUSIONS The formation of HMW fructans by individual strains of the genus Bombella is catalyzed by enzymes of the GH 32 family. Analysis of the fructan revealed an atypical structure consisting of 2,6-linked Fruf units as well as 1,2-linked Fruf units and 1,2,6-Fruf units.

Aim: Temperate phages are known to heavily impact the growth of their host, be it in a positive way, e.g., when beneficial genes are provided by the phage, or negatively when lysis occurs after prophage induction. This study provides an in-depth look into the distribution and variety of prophages in Latilactobacillus curvatus (L. curvatus). This species is found in a wide variety of ecological niches and is routinely used as a meat starter culture. Methods: Fourty five L. curvatus genomes were screened for prophages. The intact predicted prophages and their chromosomal integration loci were described. Six L. curvatus lysogens were analysed for phage-mediated lysis post induction via UV light and/or mitomycin C. Their lysates were analysed for phage particles via viral DNA sequencing and transmission electron microscopy. Results: Two hundred and six prophage sequences of any completeness were detected within L. curvatus genomes. The 50 as intact predicted prophages show high levels of genetic diversity on an intraspecies level with conserved regions mostly in the replication and head/tail gene clusters. Twelve chromosomal loci, mostly tRNA genes, were identified, where intact L. curvatus phages were integrated. The six analysed L. curvatus lysogens showed strain-dependent lysis in various degrees after induction, yet only four of their lysates appeared to contain fully assembled virions with the siphovirus morphotype. Conclusion: Our data demonstrate that L. curvatus is a (pro)phage-susceptible species, harbouring multiple intact prophages and remnant sequences thereof. This knowledge provides a basis to study phage-host interaction influencing microbial communities in food fermentations.

Objective The microbiota of a seasoning sauce fermentation process is usually complex and includes multiple species and even various strains of one species. Moreover, composition and cell numbers of individual strains vary over the course of the entire fermentation. This study demonstrates the applicability of a multiplex PCR system to monitor growth dynamics of Tetragenococcus (T.) halophilus strains in order to evaluate their performance and help to select the most competitive starter strain. Results In a previous study we isolated T. halophilus strains from multiple lupine moromi fermentation processes and characterized them. In this study we wanted to monitor the growth dynamics of these strains in a competitive lupine moromi model fermentation process using a multiplex PCR system. Therefore, pasteurized lupine koji was inoculated with eight different T. halophilus strains, six from lupine moromi, one from an experimental buckwheat moromi fermentation process and the type strain DSM 20,339 T , to create the inoculated lupine moromi pilot scale fermentation process. With the multiplex PCR system, we could detect that all strains could grow in lupine moromi but, that TMW 2.2254 and TMW 2.2264 outperformed all other strains. Both strains dominated the fermentation after three weeks with cell counts between 4 × 10 6 to 4 × 10 7 CFU/mL for TMW 2.2254 and 1 × 10 7 to 5 × 10 7 CFU/mL for TMW 2.2264. The pH dropped to value below 5 within the first 7 days, the selection of these strains might be related to their acid tolerance.

BACKGROUND:Tetragenococcus (T.) halophilus is a common member of the microbial consortia of food fermented under high salt conditions. These comprises salty condiments based on soy or lupine beans, fish sauce, shrimp paste and brined anchovies. Within these fermentations this lactic acid bacterium (LAB) is responsible for the formation of lactic and other short chain acids that contribute to the flavor and lower the pH of the product. In this study, we investigated the transcriptomic profile of the two T. halophilus strains TMW 2.2254 and TMW 2.2256 in a lupine moromi model medium supplied with galactose. To get further insights into which genomic trait is important, we used a setup with two strains. That way we can determine if strain dependent pathways contribute to the overall fitness. These strains differ in the ability to utilize L-arginine, L-aspartate, L-arabinose, D-sorbitol, glycerol, D-lactose or D-melibiose. The lupine moromi model medium is an adapted version of the regular MRS medium supplied with lupine peptone instead of casein peptone and meat extract, to simulate the amino acid availabilities in lupine moromi.RESULTS:The transcriptomic profiles of the T. halophilus strains TMW 2.2254 and TMW 2.2256 in a lupine peptone-based model media supplied with galactose, used as simulation media for a lupine seasoning sauce fermentation, were compared to the determine potentially important traits. Both strains, have a great overlap in their response to the culture conditions but some strain specific features such as the utilization of glycerol, sorbitol and arginine contribute to the overall fitness of the strain TMW 2.2256. Interestingly, although both strains have two non-identical copies of the tagatose-6P pathway and the Leloir pathway increased under the same conditions, TMW 2.2256 prefers the degradation via the tagatose-6P pathway while TMW 2.2254 does not. Furthermore, TMW 2.2256 shows an increase in pathways required for balancing out the intracellular NADH/NADH+ ratios.CONCLUSIONS:Our study reveals for the first time, that both versions of tagatose-6P pathways encoded in both strains are simultaneously active together with the Leloir pathway and contribute to the degradation of galactose. These findings will help to understand the strain dependent features that might be required for a starter strain in lupine moromi.

LebensmittelchemieVolume 77, Issue S3 p. S3-081-S3-081 Analytik Charakterisierung der von zwei Bombella-Arten aus Honigbienenstöcken gebildeten Fructane L. Ernst, L. Ernst Martin-Luther-Universität Halle-Wittenberg, Institut für Chemie - Lebensmittelchemie, Kurt-Mothes-Straße 2, 06120 Halle (S.)Search for more papers by this authorL. Härer, L. Härer Technische Universität München, Lehrstuhl für Mikrobiologie, Emil-Ramann-Str. 4, 85354 FreisingSearch for more papers by this authorJ. Bechtner, J. Bechtner Department of Food Science, Aarhus University, Nordre Ringgade 1, 8000 AarhusSearch for more papers by this authorM. Ehrmann, M. Ehrmann Technische Universität München, Lehrstuhl für Mikrobiologie, Emil-Ramann-Str. 4, 85354 FreisingSearch for more papers by this authorD. Wefers, D. Wefers Martin-Luther-Universität Halle-Wittenberg, Institut für Chemie - Lebensmittelchemie, Kurt-Mothes-Straße 2, 06120 Halle (S.)Search for more papers by this author L. Ernst, L. Ernst Martin-Luther-Universität Halle-Wittenberg, Institut für Chemie - Lebensmittelchemie, Kurt-Mothes-Straße 2, 06120 Halle (S.)Search for more papers by this authorL. Härer, L. Härer Technische Universität München, Lehrstuhl für Mikrobiologie, Emil-Ramann-Str. 4, 85354 FreisingSearch for more papers by this authorJ. Bechtner, J. Bechtner Department of Food Science, Aarhus University, Nordre Ringgade 1, 8000 AarhusSearch for more papers by this authorM. Ehrmann, M. Ehrmann Technische Universität München, Lehrstuhl für Mikrobiologie, Emil-Ramann-Str. 4, 85354 FreisingSearch for more papers by this authorD. Wefers, D. Wefers Martin-Luther-Universität Halle-Wittenberg, Institut für Chemie - Lebensmittelchemie, Kurt-Mothes-Straße 2, 06120 Halle (S.)Search for more papers by this author First published: 01 August 2023 https://doi.org/10.1002/lemi.202359062AboutPDF ToolsRequest permissionExport citationAdd to favoritesTrack citation ShareShare Give accessShare full text accessShare full-text accessPlease review our Terms and Conditions of Use and check box below to share full-text version of article.I have read and accept the Wiley Online Library Terms and Conditions of UseShareable LinkUse the link below to share a full-text version of this article with your friends and colleagues. Learn more.Copy URL Share a linkShare onEmailFacebookTwitterLinkedInRedditWechat No abstract is available for this article. References [1]L. Härer, Microorganisms 2020, 10, 1058. Volume77, IssueS3Supplement: Abstracts der Vorträge, Posterflashtalks und Poster von den 51. Deutschen Lebensmittelchemietagen 2023August 2023Pages S3-081-S3-081 ReferencesRelatedInformation

Four strains of members of the genus Bombella were isolated from samples associated with the western honey bee Apis mellifera, which could not be assigned to a species with a validly published name. Strains TMW 2.2543T, TMW 2.2556T, TMW 2.2558T and TMW 2.2559T exhibit in silico DNA-DNA hybridisation (isDDH) and orthologous average nucleotide identity (orthoANI) values below species delineation thresholds compared with all described species of the genus Bombella and with each other. TMW 2.2556T and TMW 2.2558T form their own clade within the genus. The major respiratory quinone of all strains was Q-10. The composition of cellular fatty acids was diverse between strains. All strains stained Gram-negative, were rod-shaped, strictly aerobic, pellicle-forming, catalase-positive, oxidase-negative, mesophilic and grew over a wide pH range; they were halosensitive but glucose-tolerant. Unlike the other studied strains, TMW 2.2558T was non-motile. Phylogenetic, chemotaxonomic and physiological analyses revealed a clear distinction between all the strains and species with validly published names. All the data support the proposition of four novel species within the genus Bombella, namely Bombella pluederhausensis sp. nov., Bombella pollinis sp. nov., Bombella saccharophila sp. nov. and Bombella dulcis sp. nov., with the respective type strains Bombella pluederhausensis sp. nov. TMW 2.2543T (= DSM 114872T, = LMG 32791T), Bombella pollinis sp. nov. TMW 2.2556T (= DSM 114874T, = LMG 32792T), Bombella saccharophila sp. nov. TMW 2.2558T (= DSM 114875T, = LMG 32793T) and Bombella dulcis sp. nov. TMW 2.2559T (= DSM 114877T, = LMG 32794T). Moreover, three genomes available in the NCBI database that have not yet been described as species with validly published names could be assigned to the proposed species. Bombella sp. ESL0378 and Bombella sp. ESL0385 to Bombella pollinis sp. nov. and Bombella sp. AS1 to Bombella saccharophila sp. nov.