Brewery Environment Research Articles

Understanding How Chemical Pollutants Arise and Evolve in the Brewing Supply Chain: A Scoping Review.

In this study, a critical review was carried out using the Web of ScienceTM Core Collection database to analyse the scientific literature published to date to identify lines of research and future perspectives on the presence of chemical pollutants in beer brewing. Beer is one of the world's most popular drinks and the most consumed alcoholic beverage. However, a widespread challenge with potential implications for human and animal health is the presence of physical, chemical, and/or microbiological contaminants in beer. Biogenic amines, heavy metals, mycotoxins, nitrosamines, pesticides, acrylamide, phthalates, bisphenols, microplastics, and, to a lesser extent, hydrocarbons (aliphatic chlorinated and polycyclic aromatic), carbonyls, furan-derivatives, polychlorinated biphenyls, and trihalomethanes are the main chemical pollutants found during the beer brewing process. Pollution sources include raw materials, technological process steps, the brewery environment, and packaging materials. Different chemical pollutants have been found during the beer brewing process, from barley to beer. Brewing steps such as steeping, kilning, mashing, boiling, fermentation, and clarification are critical in reducing the levels of many of these pollutants. As a result, their residual levels are usually below the maximum levels allowed by international regulations. Therefore, this work was aimed at assessing how chemical pollutants appear and evolve in the brewing process, according to research developed in the last few decades.

Genomic Analysis and In Vitro Investigation of the Hop Resistance Phenotype of Two Novel Loigolactobacillus backii Strains, Isolated from Spoiled Beer.

Loigolactobacillus backii is an important beer-spoiling species, exhibiting high hop tolerance. Here, we present the annotated whole genome sequence of two recently isolated strains, Lg. backii KKP 3565 and KKP 3566. Firstly, to study the genetic basis of the persistence of the two isolates in beer, a comprehensive bioinformatic analysis ensued. Their chromosome map was constructed, using whole-genome sequencing and assembly, revealing that the two strains carry genomes with a length of 2.79 Mb with a GC content of 40.68%. An average nucleotide identity (ANI) analysis demonstrated that the novel strains possess unique genomic sequences, also confirming their classification into the Lg. backii species. Their genome harbors numerous insertion sequences and plasmids, originating from other beer-spoiling species. Regarding their adaptation in brewery environment, homologous genes that confer resistance to hop were spotted, while the impact of hop bitters and pure beer on bacterial growth was investigated, in vitro. In brief, low hop concentrations were found to induce the proliferation of strains, while a higher concentration negatively affected their growth. Nonetheless, their ability to survive in pure beer indicated their tolerance to high hop concentrations. These results offer insight into the capacity of Lg. backii KKP 3566 and Lg. backii KKP 3566 to tolerate the extreme conditions prevalent in the brewery environment.

Development of a Rapid and Accurate Nanopore-based Sequencing Platform for on-Field Identification of Beer-Spoilage Bacteria in the Breweries

When a beer is contaminated with unknown microorganisms, rapid and accurate identification of the contaminants and risk assessment of their spoilage capability are of critical importance for proper corrective actions. Molecular-based identification techniques have been used for years to identify beer-spoilage bacteria in the breweries. However, these techniques require expensive, advanced analytical instruments (e.g., DNA sequencer, MALDI-TOF MS, etc.), and advanced skills, hindering their practical use for routine microbiological testing in the field. Thus, the evaluation and development of alternative methods for microbial identification that are low-cost, but rapid and provide the same level of accuracy, are a necessity. One particular technology is the nanopore-based sequencer, MinION, developed by Oxford Nanopore Technologies (ONT). MinION is a low cost, portable real-time device for DNA and RNA sequencing. We developed and evaluated a MinION sequencer-based platform for beer-spoilage bacterial identification. The near full length 16S rRNA gene of a wide variety of beer-spoilage bacteria and those commonly found in the brewery environment, were sequenced using three different sequencing kits and analyzed using the EPI2ME cloud-based analysis platform and a consensus sequence-based approach. The methods successfully rapidly identified test-microorganisms at a species level, (approx. 3.5 h from DNA extraction to data analysis) and with a high accuracy, similar to Sanger sequencing (average per. identity > 98.5%). The MinION sequencer-based microbial identification platform was shown to be a powerful tool for on-field rapid and accurate beer-spoilage bacterial identification, which could revolutionize the way microbial identification is performed in a brewery QC laboratory.

Nanopore based sequencing enables easy and accurate identification of yeasts in breweries

In breweries, rapid and accurate identification of unknown microbial contaminants and the associated risk assessment of beer spoilage are of critical importance to ensure proper corrective measures are taken. In this context, the nanopore based sequencer, MinION, developed by Oxford Nanopore Technologies, is of particular interest. MinION is a low cost, portable, real time device for DNA and RNA sequencing. Recently, our group developed and evaluated a MinION sequencer based microbial identification platform for beer spoilage bacteria. In this study, it is used to identify yeast strains. The nuclear ribosomal RNA genes and their internal transcribed spacer regions from a wide variety of beer spoilage yeasts, commonly found in the brewery environment, were sequenced using two different sequencing kits - Ligation Sequencing Kit and Rapid Barcoding Kit - and analysed using consensus sequence-based approach. Comparative evaluation of Sanger sequencing and MinION sequencing was performed to evaluate the performance of the MinION platform in the identification of yeasts. Our methods successfully identified yeasts at the species level rapidly (approximately 3.5 hours from DNA extraction to data analysis), and with high accuracy that is comparable to Sanger sequencing (average percentage identity >98.9%). The MinION sequencer based microbial identification platform, equipped for both bacterial and yeast identification, is a powerful tool for on-field rapid and accurate identification of beer spoilage and brewery related microorganisms. This new identification approach will potentially revolutionise the way microbial identification is performed in brewery quality control and assurance laboratories. © 2021 The Institute of Brewing & Distilling

Combined Yeast Biofilm Screening – Characterization and Validation of Yeast Related Biofilms in a Brewing Environment with Combined Cultivation and Specific Real-time PCR Screening of Selected Indicator Species

Microbial spoilage of alcohol-free and low-alcohol beers, beer-mixed beverages, and soft drinks is most commonly caused by yeast. Yeast-related biofilms are therefore a serious problem in the production of these beverages. Fast detection of developing biofilms is a key factor to prevent subsequent spoilage of the product. For fast yeast detection, a new specific medium was developed and combined with real-time polymerase chain reaction (PCR) detection of characteristic beverage spoiling yeast species. The medium is based on MYPG broth (malt extract, yeast extract, peptone, glucose broth) with resazurin as a redox indicator for cell activity. The growth and biofilm potential of representative strains of commonly present beverage spoilage yeast species was evaluated using the developed medium. A novel real-time PCR detection system for Rhodotorula mucilaginosa, an early biofilm colonizer, was designed and successfully validated. Two field tests of the medium in combination with real-time PCR were performed. One test showed a differentiated hygienic status on a filler, and the other test tracked the contamination source of Saccharomyces cerevisiae var. diastaticus. Biofilm relevance of the strain set was proven. The modified MYPG proved to be highly sensitive when detecting yeasts. The detection of the selected target species directly in the medium was compatible and can provide detailed hygienic profiles when combined with additional information on the target species. This provides a fast detection method for yeast-related biofilms in brewery environments, differentiated hygienic monitoring, and makes it possible to troubleshoot contamination incidents.

Coastal sand dune ecosystem services in metropolitan suburbs: effects on the sake brewery environment induced by changing social conditions

Chiba Prefecture, Japan, lies very near Tokyo, the capital city of Japan. It borders the sea on three sides and is banded by coastal dunes. Several sake breweries are located near these dunes. Although there are records of sake brewing along the coast of Tokyo Bay since 1925, sake breweries have completely disappeared in several areas. We believe that sake brewing in these areas benefited from the ecosystem services afforded them by their proximity to the coastal ecosystem. We investigated potential environmental factors (e.g., landscape, soil, and groundwater), as well as conditions that could have driven sake brewers away from the coastal area. Many of the sake breweries that no longer exist were located on coastal dunes (i.e., sand, sandbanks, and natural levees) and obtained their water from a freshwater layer located 3–10 m below the surface. We speculate that these sake breweries benefited from using natural ingredients found in the coastal zone. We also investigated the following factors that may have negatively impacted the breweries, driving them out of business: (1) bankruptcies and reconstruction difficulties that followed the destructive 1923 Great Kanto earthquake, (2) industrial wartime adjustments during World War II (1939–1945), (3) development of coastal industries during the period from 1960 to 1975, and (4) increasing choices for other alcoholic drinks (e.g., beer, wine, and whiskey) from the 1960s to the present.

Distribution of Lactobacillus and Pediococcus in a Brewery Environment

Beer is a fermented beverage in which microbial activity is central to its production, defining the characteristics of the final product. The diversity of microorganisms present in this process is, however, relatively limited, mainly owing to its restrictive environment, such as the iso-α-acids from hops and ethanol, which have antimicrobial activity. Despite these unfavorable conditions for bacterial growth, some beer spoilers are able to grow in beer. Lactic acid bacteria (LAB) are reported to be responsible for approximately 70% of all beer-spoilage incidents. These beer spoilers have the common characteristic of resistance to iso-α-acids found in hops given by genetic elements such as ORF5 and the multidrug transporter horA. However, the tolerance to hop compounds could be a multifactorial trait that includes the bacterial species and the hop-resistance genes. So it is important to track the bacterial beer-spoiler species and hop-resistance genes in a brewery environment. In this work we isolated 10 beer-spoiling LAB strains and one Bacillus sp. from a commercial brewery environment, and we found hop-resistance genes in nine out of the 10 LAB strains and also in the non–beer spoiler Bacillus. The isolated LAB strains were identified by rRNA 16S gene sequence and digestion with the restriction enzyme BfaI, resulting in four species, Lactobacillus rossiae, Pediococcus damnosus, Lactobacillus brevis, and Lactobacillus casei. Most of the strains and hop-resistance genes were found in the environment of the fermentation stage.

First study on the formation and resuscitation of viable but nonculturable state and beer spoilage capability of Lactobacillus lindneri

This study aimed to investigate the spoilage capability of Lactobacillus lindneri during the induction and resuscitation of viable but nonculturable (VBNC) state. L.lindneri strain was identified by sequencing the PCR product (amplifying 16S rRNA gene) using ABI Prism 377 DNA Sequencer. During the VBNC state induction by low temperature storage and beer adaption, total, culturable, and viable cells were assessed by acridine orange direct counting, plate counting, and Live/Dead BacLight bacterial viability kit, respectively. Organic acids and diacetyl concentration were measured by reversed-phase high performance liquid chromatography and head dpace gas chromatography, respectively. VBNC state of L.lindneri was successfully induced by both beer adaption and low temperature storage, and glycerol frozen stock was the optimal way to maintain the VBNC state. Addition of catalase was found to be an effective method for the resuscitation of VBNC L.lindneri cells. Furthermore, spoilage capability remained similar during the induction and resuscitation of VBNC L.lindneri. This is the first report of induction by low temperature storage and resuscitation of VBNC L.lindneri strain, as well as the first identification of spoilage capability of VBNC and resuscitated L.lindneri cells. This study indicated that the potential colonization of L.lindneri strain in brewery environment, formation and resuscitation of VBNC state, as well as maintenance in beer spoilage capability, may be an important risk factor for brewery environment.

Pediococcus damnosusstrains isolated from a brewery environment carry thehorAgene

Beer is recognized as a safe beverage, owing to its excellent microbiological stability provided by its components, especially iso-α-acids from hop and ethanol which have antimicrobial activity. Despite these unfavourable conditions for bacteria, some lactic acid bacteria (LAB) can cause beer spoilage. Resistance to hop compounds is caused, in part, by the product of genes like horA. Understanding how LAB adapts to hop compounds as well as quick detection of these microorganisms is necessary to ensure high-quality beverages produced by the brewing industry. In this work, we searched for the presence of two main hop-resistance genes, horA and ORF5, and determined the capacity of four strains of Pediococcus damnosus isolated from a brewery environment to grow in the presence of increasing concentrations of iso-α-acids. All strains were able to grow in increasing concentrations of iso-α-acids up to 150 μg mL−1. This amount is 10 times greater than the concentration in average beer. Genetic amplification of genes associated with hop-resistance, demonstrated the presence of horA, but not ORF5 in all tested strains. This communication represents the first report of the presence of horA gene in bacteria isolated from breweries in our country. Copyright © 2017 The Institute of Brewing & Distilling

Review: Gram Negative Bacteria in Brewing

Gram negative aerobic bacteria such as Acetic Acid Bacteria, which include Acetobacter and Gluconobacter, have historically caused significant problems to brewers. Although incidences of spoilage have recently reduced as a result of improvements in beer packaging, these bacteria are still a concern in dispense systems in pub breweries, public houses and cask conditioned beers. Gram negative facultative bacteria of the genus Zymomonas can spoil primed cask conditioned beer and cider. There is a wide range of Enterobacteraeceace which are found within brewery environments and they serve as indicator microorganisms for hygiene and sanitation. Gram negative strictly anaerobic bacteria such as Pectinatus and Megasphaera have recently emerged as a significant threat due to the improvement in reduction of oxygen levels in beer and an increase in production of unpasteurised beer. Pectinatus and Megasphaera are sensitive to routine cleaning agents used in breweries, but they can survive and proliferate in biofilms eventually causing spoilage of beer. This review focuses on Gram negative aerobic, facultative anaerobic and strictly anaerobic brewery related spoilage bacteria.

Nisin-producing microorganisms and their implementation in brewers' wort

Nisin is a bacteriocin, which is capable of eliminating more than 90% of all potential beer spoilage Gram-positive bacteria. Hence, the implementation of nisin-producing cultures into the brewing process needs to be evaluated systematically. In this work, the genetic relationships and properties, as well as the reactions of four strains of Lactococcus lactis ssp. lactis, known to be capable of producing nisin (NCIMB 8780, 8586, 701402 and 701403), and seven isolated strains of the same species found in the beverage environment (TUM 575, 8947, 8127, 8446, 8673, 8973 and 8872), were tested under typical brewery conditions. As in previous work, it was found, that all of the tested strains could be genetically differentiated via PCR-(GTG)5. In addition, the absence of the genes HorA, HorC and ORF5 indicated that all strains were sensitive to hop components. The agar diffusion assay test proved to be the most reliable method to precisely determine different nisin concentrations. Nisin formation, acid formation and the reproduction rates of the organisms were tested subsequently in various brewery relevant culture media such as MRS, first wort (unhopped) and wort (hopped). MRS provided the best environment for bacterial growth and hence acid and nisin production. The four NCIMB strains, which were the only ones capable of producing nisin under the named conditions, were chosen for study with regards to their tolerance to specific compounds found in the brewery environment. The strains NCIMB 8780 and 8586 produced comparatively higher amounts of nisin and were more tolerant to hop bitterness, ethanol, high gravity and low extract conditions. The growth rates, acid production and nisin production of all strains decreased with increasing bitter units and ethanol content. The optimum extract concentration was 5–10°P. Copyright © 2015 The Institute of Brewing & Distilling

Nisin and its usage in breweries: a review and discussion

The most common source of microbiological contamination in today's brewing industry is Gram-positive bacteria. Therefore, different approaches are used in daily quality assurance routines in order to prevent and to eliminate infections. Normally, common disinfection agents are applied within the automated cleaning processes, but there is a growing market requirement for the reduction of their usage and a substitution with natural ingredients. Therefore, implementation of a natural bio-preservative such as nisin, a bacteriocin capable of eliminating up to 90% of Gram-positive beer spoilage bacteria, poses an effective, natural solution. This article summarizes bacteriocins currently used in food production. Subsequent specific details about nisin including its mode of action, characteristics, legal aspects, production methods, interferences and synergistic factors are presented. Additionally, a variety of analytical methods to determine nisin's concentration are included and possible industrial applications of nisin in the brewery environment are discussed. Copyright © 2015 The Institute of Brewing & Distilling

Reduction and restoration of culturability of beer-stressed and low-temperature-stressed Lactobacillus acetotolerans strain 2011-8

Lactic acid bacteria (LAB) are the most common beer-spoilage bacteria, regardless of beer type, and therefore pose significant problems for the brewing industry. The aim of this study was to investigate the viable, but putatively non-culturable (VPNC) state of the hard-to-culture beer-spoilage species, Lactobacillus acetotolerans. Upon prolonged contact with degassed beer, L. acetotolerans was found to show decreased culturability. After 17 subcultures in beer, 100-μL aliquots of the culture were no longer culturable on MRS agar until 14days of incubation despite the presence of 105 viable cells, indicating that a large population of cells entered into a VPNC state. Furthermore, a significant reduction or even putative loss of culturability, but maintenance of viability, of L. acetotolerans could also be induced by storing the strain at 0°C for 105days. Adding catalase at a concentration of 1000U/plate enabled the VPNC cells, both induced by beer subculture treatment and cold treatment, to regain culturability with a resuscitation time of 4days and 3days, respectively. Scanning electron microscopy results demonstrated that cells decreased in size and gradually changed morphology from short rods to coccoids when they entered the VPNC state. It was concluded that the difficulty in culturing the spoilage bacterium from brewery environments could be partly attributed the hard-to-culture or the viable, but non-culturable characteristic of this organism.

Neural Virtual Sensors for Adaptive Magnetic Localization of Autonomous Dataloggers

The surging advance in micro- and nanotechnologies allied with neural learning systems allows the realization of miniaturized yet extremely powerful multisensor systems and networks for wide application fields, for example, in measurement, instrumentation, automation, and smart environments. Time and location context is particularly relevant to sensor swarms applied for distributed measurement in industrial environment, such as, for example, fermentation tanks. Common RF solutions face limits here, which can be overcome by magnetic systems. Previously, we have developed the electronic system for an integrated data logger swarm with magnetic localization and sensor node timebase synchronization. The focus of this work is on an approach to improving both localization accuracy and flexibility by the application of artificial neural networks applied as virtual sensors and classifiers in a hybrid dedicated learning system. Including also data from an industrial brewery environment, the best investigated neural virtual sensor approach has achieved an advance in localization accuracy of a factor of 4 compared to state-of-the-art numerical methods and, thus, results in the order of less than 5 cm meeting industrial expectations on a feasible solution for the presented integrated localization system solution.

MALDI-TOF MS Analysis of Anaerobic Bacteria Isolated from Biofilm-Covered Surfaces in Brewery Bottling Halls

Brewery filling equipment is continually exposed to moisture and nutrients during production runs and is therefore susceptible to microbial attachment and biofilm development. These biofilms play the role of reservoirs of beer spoilage microorganisms. We evaluated the usability of matrix assisted laser desorption/ionization time-of-flight mass spectrometry (MALDI-TOF MS) analysis for direct identification of beer contaminants and direct analysis of biofilm. From a total of 58 isolates obtained from the beer filling hall environment, 47% were successfully assigned to the species level and 10% to the genus level when using the commercially available BioTyper (Bruker Daltonik) database. After homemade extension of the database by eight entries belonging to genera Pectinatus, Megasphaera, and Selenomonas, the identification success rate at the species level was increased to 85%. The possibility of direct biofilm analysis was also examined by testing four sample preparation procedures; however, identification of beer contaminants was not successful, as protein signals needed for microorganism identification were not detected by this approach. MALDI-TOF MS including the cultivation step in connection with a reference database containing relevant entries was found as a suitable method for identification of contaminants from brewery environment.

Gluconobacter cerevisiae sp. nov., isolated from the brewery environment

Three strains, LMG 27748(T), LMG 27749 and LMG 27882 with identical MALDI-TOF mass spectra were isolated from samples taken from the brewery environment. Analysis of the 16S rRNA gene sequence of strain LMG 27748(T) revealed that the taxon it represents was closely related to type strains of the species Gluconobacter albidus (100 % sequence similarity), Gluconobacter kondonii (99.9 %), Gluconobacter sphaericus (99.9 %) and Gluconobacter kanchanaburiensis (99.5 %). DNA-DNA hybridization experiments on the type strains of these species revealed moderate DNA relatedness values (39-65 %). The three strains used d-fructose, d-sorbitol, meso-erythritol, glycerol, l-sorbose, ethanol (weakly), sucrose and raffinose as a sole carbon source for growth (weak growth on the latter two carbon sources was obtained for strains LMG 27748(T) and LMG 27882). The strains were unable to grow on glucose-yeast extract medium at 37 °C. They produced acid from meso-erythritol and sucrose, but not from raffinose. d-Gluconic acid, 2-keto-d-gluconic acid and 5-keto-d-gluconic acid were produced from d-glucose, but not 2,5-diketo-d-gluconic acid. These genotypic and phenotypic characteristics distinguish strains LMG 27748(T), LMG 27749 and LMG 27882 from species of the genus Gluconobacter with validly published names and, therefore, we propose classifying them formally as representatives of a novel species, Gluconobacter cerevisiae sp. nov., with LMG 27748(T) ( = DSM 27644(T)) as the type strain.

Hydrophobins: Exceptional proteins for many applications in brewery environment and other bio-industries

Hydrophobins are exceptional proteins produced by fungi. Research over the last decade has led to a better understanding of their role in spontaneous self-assembly at hydrophobic/hydrophilic interfaces. This has resulted in many proposals for using hydrophobins in many important scientific and technological applications. Hydrophobins may become attractive as special biosurfactants, as foaming agents and for protein immobilization in the food industries and in biosensors. Moreover, they can be interesting as stabilizers for flavors, and as, encapsulating agents of trace ingredients in beverages. The use of hydrophobins in pharmaceutical formulations and in medicine is another interesting application as they cause an increased stabilization of drugs. The study of hydrophobins must also lead to a better understanding of the gushing phenomenon in beverages like beers, wines and ciders, which causes great economic losses in those fields. To recognize the positive and the negative aspects of hydrophobins these proteins should be commercially available at large scale which however is not the case. An overview of existing possibilities for applications may help to understand their behavior in different environmental conditions and to stimulate finding improved methods for isolation and purification, and possibly other unexpected applications.

Occurrence of Pectinatus and Megasphaera in the Major UK Breweries

The occurrence of beer spoilage bacteria belonging to the genera Pectinatus and Megasphaera in ten major UK breweries was investigated. The sampling points were selected from fermentation areas, beer conditioning areas and beer bottling and canning sites. Multiplex PCR methodology was used for detection of three Pectinatus and three Megasphaera species using species-specific primers. The presence of six Lactobacillus species was also examined. Overall, 117 samples were analysed from ten breweries; six samples were positive for the presence of Pectinatus species and three samples were positive for the presence of Megasphaera species, while 34 samples were positive for the presence of Lactobacillus species. Lactobacillus species appeared to be the major potential spoilage microorganisms. Although none of the actual beer samples were found to be positive for Pectinatus and Megasphaera species, their occurrence in aerobic brewery environments indicates sanitation problems and revealed the presence of highly established biofilms in some breweries.

Aspects of <i>Garcina kola</i> Toxicity to a Wild Strain of <i>Saccharomyces cerevisiae</i> from a Brewery Environment

Bitter cola (Garcinia kola) powder and its ethanolic extract were assessed for their ability to inhibit the growth of a wild strain of Saccharomyces cerevisiae isolated from a brewery environment. The iso-alpha acid contents of the G. kola powder, G. kola extract and hop extract were determined as 5.85 mg/l, 6.34 mg/l and 7.65 mg/l, respectively, showing that the iso-alpha acid contents of G. kola extract and G. kola powder compared relatively well with that of hop extract. The antimicrobial activity of both agents, using Humulus lupulus ethanolic extract as control, showed a high dependence on the temperature of assay. At most of the temperatures evaluated (30 – 45 oC), the rates of antimicrobial activity differed. Garcinia kola ethanolic extract and hops extract displayed total inhibition of the wild yeast growth at temperature of 40 oC at concentrations of about 0.039 mg/ml and 0.078 mg/ml respectively. Humulus lupulus extract (hops) had the highest generation time at 450C at a concentration of 0.078 mg/ml. The complete inhibition of the yeast growth by G. kola extract at 40 oC indicated that it had higher antimicrobial activity against this yeast than hops extract. In all the temperatures evaluated both G. kola ethanolic extract and powder compared favourably with hops extract with respect to antimicrobial activity against a wild strain of Saccharomyces cerevisiae at various concentrations. These results revealed that the antimicrobial activity of these hopping agents were temperature and concentration dependent.

Lactobacillus paucivorans sp. nov., isolated from a brewery environment

A Gram-stain-positive, catalase-negative and rod-shaped bacterium was isolated from a brewery environment. Its phylogenetic affiliation was determined by using 16S rRNA gene sequence analysis. It was found that strain TMW 1.1424(T) belongs to the genus Lactobacillus, with the three nearest neighbours Lactobacillus parabrevis LMG 11984(T) (97 %), Lactobacillus brevis DSM 20054(T) (95.9 %) and Lactobacillus hammesii DSM 16381(T) (96.2 %). Comparative sequencing of additional phylogenetic marker genes tuf and pheS confirmed the 16S rRNA gene sequence tree topology. The DNA G+C content of strain TMW 1.1424(T) is 46.6 mol%. Genomic DNA-DNA relatedness values with L. brevis DSM 20054(T), L. parabrevis LMG 11984(T) and L. hammesii DSM 16381(T) do not exceed 52.8 %, revealing that the novel isolate represents a separate genomic species. The strain can be distinguished from other related species of the genus Lactobacillus by physiological and biochemical tests. Based on biochemical, physiological and phylogenetic data, it is proposed that the new isolate be classified as a novel species of the genus Lactobacillus, Lactobacillus paucivorans sp. nov. The type strain is TMW 1.1424(T) (=DSM 22467(T) =LMG 25291(T)).