Fast growth can counteract antibiotic susceptibility in shaping microbial community resilience to antibiotics.

Microbial communities often face external perturbations that can induce lasting changes in their composition and functions. Our understanding of how multispecies communities respond to perturbations such as antibiotics is limited, with susceptibility assays performed on individual, isolated species our primary guide in predicting community transitions. Here, we studied how bacterial growth dynamics can overcome differences in antibiotic susceptibility in determining community resilience: the recovery of the original community state following antibiotic exposure. We used an experimental community containing Corynebacterium ammoniagenes and Lactobacillus plantarum that displays two alternative stable states as a result of mutual inhibition. Although C. ammoniagenes was more susceptible to chloramphenicol in monocultures, we found that chloramphenicol exposure nonetheless led to a transition from the L. plantarum-dominated to the C. ammoniagenes-dominated community state. Combining theory and experiments, we demonstrated that growth rate differences between the two species made the L. plantarum-dominated community less resilient to several antibiotics with different mechanisms of action. Taking advantage of an observed cooperativity­a dependence on population abundance­in the growth of C. ammoniagenes, we next analyzed in silico scenarios that could compromise the high resilience of the C. ammoniagenes-dominated state. The model predicted that lowering the dispersal rate, through interacting with the growth at low population densities, could make the C. ammoniagenes state fragile against virtually any kind of antibiotic, a prediction that we confirmed experimentally. Our results highlight that species susceptibility to antibiotics is often uninformative of community resilience, as growth dynamics in the wake of antibiotic exposure can play a dominant role.

Symbiotic microbiome Staphylococcus aureus from human nasal mucus modulates IL-33-mediated type 2 immune responses in allergic nasal mucosa.

BACKGROUND: The host-microbial commensalism can shape the innate immune responses in respiratory mucosa and nasal microbiome also modulates front-line immune mechanism in the nasal mucosa. Inhaled allergens encounter the host immune system first in the nasal mucosa, and microbial characteristics of nasal mucus directly impact the mechanisms of initial allergic responses in nasal epithelium. However, the roles of the nasal microbiome in allergic nasal mucosa remain uncertain. We sought to determine the distribution of nasal microbiomes in allergic nasal mucosa and elucidate the interplay between nasal microbiome Staphylococcus species and Th2 cytokines in allergic rhinitis (AR) models. RESULTS: Staphylococcus aureus (AR-SA) and S. epidermidis (AR-SE) were isolated from the nasal mucosa of patients with AR. The influence of nasal microbiome Staphylococcus species on allergic nasal mucosa was also tested with in vitro and in vivo AR models. Pyrosequencing data showed that colonization by S. epidermidis and S. aureus was more dominant in nasal mucus of AR subjects. The mRNA and protein levels of IL-33 and TSLP were significantly higher in AR nasal epithelial (ARNE) cells which were cultured from nasal mucosa of AR subjects, and exposure of ARNE cells to AR-SA reduced IL-33 mRNA and secreted protein levels. Particularly, ovalbumin-driven AR mice inoculated with AR-SA by intranasal delivery exhibited significantly reduced IL-33 in their nasal mucosa. In the context of these results, allergic symptoms and Th2 cytokine levels were significantly downregulated after intranasal inoculation of AR-SA in vivo AR mice. CONCLUSION: Colonization by Staphylococcus species was more dominant in allergic nasal mucosa, and nasal commensal S. aureus from subjects with AR mediates anti-allergic effects by modulating IL-33-dependent Th2 inflammation. The results demonstrate the role of host-bacterial commensalism in shaping human allergic inflammation.

Zinc Oxide Inhibits Axillary Colonization by Members of the Genus Corynebacterium and Attenuates Self-perceived Malodour: A Randomized, Double-blind, Placebo-controlled Trial.

Malodour from the axilla is commonly caused by specific microbes, and may be inhibited by zinc oxide. The aim of this study was to determine the effects of zinc oxide on the axillary microbiota, odour and pH in a randomized, double-blind, placebo-controlled trial in 30 healthy volunteers. In each participant 1 axilla was treated with zinc oxide and the other with a placebo for 13 days. The microbiota and pH were analysed before and during treatment. At the final visit, the participants judged their own axillary odour for comparison. With zinc oxide treatment total bacterial growth and, specifically, that of odour-producing Corynebacterium spp. and Staphylococcus hominis, decreased (p < 0.05), despite an increase (p < 0.0005) in skin-surface pH. Compared with the placebo, zinc oxide treatment reduced (p = 0.005) self-perceived malodour. In vitro, Corynebacterium spp. (19 isolated strains) survival was reduced (p < 0.0005) at pH 5.0 compared with pH 6.0; growth inhibition by zinc oxide occurred at ≤ 400 mg/l, and cell death occurred at ≤ 10,000 mg/l for 12 (63%) of the strains. In conclusion, application of zinc oxide reduced malodour and the counts of causative bacteria, but increased the pH of the axilla.

Reconstructing the Phylogeny of Corynebacteriales while Accounting for Horizontal Gene Transfer.

Horizontal gene transfer is a common mechanism in Bacteria that has contributed to the genomic content of existing organisms. Traditional methods for estimating bacterial phylogeny, however, assume only vertical inheritance in the evolution of homologous genes, which may result in errors in the estimated phylogenies. We present a new method for estimating bacterial phylogeny that accounts for the presence of genes acquired by horizontal gene transfer between genomes. The method identifies and corrects putative transferred genes in gene families, before applying a gene tree-based summary method to estimate bacterial species trees. The method was applied to estimate the phylogeny of the order Corynebacteriales, which is the largest clade in the phylum Actinobacteria. We report a collection of 14 phylogenetic trees on 360 Corynebacteriales genomes. All estimated trees display each genus as a monophyletic clade. The trees also display several relationships proposed by past studies, as well as new relevant relationships between and within the main genera of Corynebacteriales: Corynebacterium, Mycobacterium, Nocardia, Rhodococcus, and Gordonia. An implementation of the method in Python is available on GitHub at https://github.com/UdeS-CoBIUS/EXECT (last accessed April 2, 2020).

Of microbes and mange: consistent changes in the skin microbiome of three canid species infected with Sarcoptes scabiei mites.

BACKGROUND: Sarcoptic mange is a highly contagious skin disease caused by the ectoparasitic mite Sarcoptes scabiei. Although it afflicts over 100 mammal species worldwide, sarcoptic mange remains a disease obscured by variability at the individual, population and species levels. Amid this variability, it is critical to identify consistent drivers of morbidity, particularly at the skin barrier. METHODS: Using culture-independent next generation sequencing, we characterized the skin microbiome of three species of North American canids: coyotes (Canis latrans), red foxes (Vulpes vulpes) and gray foxes (Urocyon cinereoargenteus). We compared alpha and beta diversity between mange-infected and uninfected canids using the Kruskal-Wallis test and multivariate analysis of variance with permutation. We used analysis of composition of microbes and gneiss balances to perform differential abundance testing between infection groups. RESULTS: We found remarkably consistent signatures of microbial dysbiosis associated with mange infection. Across genera, mange-infected canids exhibited reduced microbial diversity, altered community composition and increased abundance of opportunistic pathogens. The primary bacteria comprising secondary infections were Staphylococcus pseudintermedius, previously associated with canid ear and skin infections, and Corynebacterium spp., previously found among the gut flora of S. scabiei mites and hematophagous arthropods. CONCLUSIONS: This evidence suggests that sarcoptic mange infection consistently alters the canid skin microbiome and facilitates secondary bacterial infection, as seen in humans and other mammals infected with S. scabiei mites. These results provide valuable insights into the pathogenesis of mange at the skin barrier of North American canids and can inspire novel treatment strategies. By adopting a "One Health" framework that considers mites, microbes and the potential for interspecies transmission, we can better elucidate the patterns and processes underlying this ubiquitous and enigmatic disease.

Subacute Osteomyelitis of the Fibula due to Corynebacterium striatum in an Immunocompetent Child: A Case Report.

CASE: A 4-year-old boy presented with a history of swelling and pain in his ankle for 1 month. X-ray showed a periosteal reaction, and magnetic resonance imaging (MRI) favored radiodense signs in a T2-weighted image. An open biopsy was performed, and cultures grew Corynebacterium spp. Further research showed that the pathogen was Corynebacterium striatum subtype. Vancomycin and cefazolin were given for 6 weeks. Clinical, X-ray, and MRI signs indicated improvement, and there were no complications with 2-year follow-up. CONCLUSIONS: C. striatum is an unexpected pathogen for subacute osteomyelitis in children. As far as we know, this is the first case report of subacute osteomyelitis in a child with a normal immune system caused by C. striatum.

Virulence potential of Corynebacterium striatum towards Caenorhabditis elegans.

Corynebacterium striatum strains have been increasingly reported as etiological agents of nosocomial infections and outbreaks in industrialized and developing countries. However, there are few studies focused on the virulence potential of C. striatum. A growing body of research supports the use of Caenorhabditis elegans as a model host for investigating the virulence potential of pathogenic bacteria, including corynebacteria. In the present study, chemotaxis behaviour, mortality, and morphological changes were investigated in nematodes infected by four C. striatum strains isolated from different clinical sites, and with different MDR profiles and PFGE types. The results showed chemotaxis of nematodes towards C. striatum. Nematode death (> 60%) was detected from the first day post-infection with all strains tested, but at different levels, independent of biofilm formation on catheter surfaces and differences in growth temperature between nematodes (20 °C) and mammals (37 °C). C. striatum 2369/II multidrug-resistant (MDR; from tracheal aspirate of a patient undergoing endotracheal intubation) and 1961/III multidrug-sensitive (MDS; urine) strains led to 100% mortality in worms. Survival of nematodes was observed until 4 days post-infection with the C. striatum 1954/IV MDS strain isolated from a surgical wound (13%) and 1987/I MDR strain isolated from a patient with a lower respiratory tract infection (39%). The Dar phenotype was observed post-infection with all MDS and MDR strains except 1954/IV. All strains showed the capacity for bagging formation. Star formation was observed only with strains that led to 100% nematode mortality. In conclusion, C. striatum was found to exert virulence for C. elegans. Variations in nematode morphological changes and levels of mortality indicate differences in the virulence potential of C. striatum independent of clinical isolation site, capacity for biofilm formation, and MDR and PFGE profiles.

Biological ball filters regulate bacterial communities in marron (Cherax cainii) culture system.

This study aimed to characterize the bacterial communities in rearing water treated with commercial plastic biological ball filters named as Bio-ball in marron culture for 60 days. Inclusion of Bio-ball in the aquaculture tanks showed improvement in water quality parameters and enrichment of bacterial communities in terms of operational taxonomic units. The water treated with Bio-ball showed significantly less nitrate, nitrite, ammonia, phosphorus and high dissolved oxygen concentration than untreated control group. At phylum level, Proteobacteria was dominant in both control and treated water, whereas Firmicutes was found to be significantly (P < 0·05) enriched in Bio-ball treated water. Among the classified genus, Aquabacterium and Polunucleobacter were most dominant in control and Bio-ball treated water respectively. Linear Discriminant Analysis Effect Size exhibited 31 indicator bacterial genus, 10 in control and 21 in treated condition, suggesting the enrichment of microbial lineages with addition of Bio-ball. The bacteria Haliscomenobacter, Hypnocyclicus, Pajaroellobacter and Vibrio were found to be significantly (P < 0·001) correlated with higher pH, nitrate, nitrite, phosphorus and ammonia in control tanks, whereas Corynebacterium was linked to higher temperature in treated water. Overall results suggest that Bio-ball filter media significantly improved the water quality and microbial populations in aquaculture tanks. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of this study revealed the positive impacts of Bio-ball in enrichment of microbial flora associated with the degradation process of nitrogenous and organic compounds. Bio-ball also showed the capability to prevent the colonization of harmful bacteria, and favoured the growth of beneficial microbes in aquatic system. This study therefore could pave the ways of increasing the aquaculture production by improving the water quality.

Corynebacterium pseudodiphtheriticum Exploits Staphylococcus aureus Virulence Components in a Novel Polymicrobial Defense Strategy.

Commensal bacteria in the human nasal cavity are known to suppress opportunistic pathogen colonization by competing for limited space and nutrients. It has become increasingly apparent that some commensal bacteria also produce toxic compounds that directly inhibit or kill incoming competitors. Numerous studies suggest that microbial species-specific interactions can affect human nasal colonization by the opportunistic pathogen Staphylococcus aureus However, the complex and dynamic molecular interactions that mediate these effects on S. aureus nasal colonization are often difficult to study and remain poorly understood. Here, we show that Corynebacterium pseudodiphtheriticum, a common member of the normal nasal microbiota, mediates contact-independent bactericidal activity against S. aureus, including methicillin-resistant S. aureus (MRSA). Bacterial interaction assays revealed that S. aureus isolates that were spontaneously resistant to C. pseudodiphtheriticum killing could be recovered at a low frequency. To better understand the pathways associated with killing and resistance, a S. aureus transposon mutant library was utilized to select for resistant mutant strains. We found that insertional inactivation of agrC, which codes for the sensor kinase of the Agr quorum sensing (Agr QS) system that regulates expression of many virulence factors in S. aureus, conferred resistance to killing. Analysis of the spontaneously resistant S. aureus isolates revealed that each showed decreased expression of the Agr QS components. Targeted analysis of pathways regulated by Agr QS revealed that loss of the phenol-soluble modulins (PSMs), which are effectors of Agr QS, also conferred resistance to bactericidal activity. Transmission electron microscopy analysis revealed that C. pseudodiphtheriticum induced dramatic changes to S. aureus cell surface morphology that likely resulted in cell lysis. Taken together, these data suggest that C. pseudodiphtheriticum-mediated killing of S. aureus requires S. aureus virulence components. While S. aureus can overcome targeted killing, this occurs at the cost of attenuated virulence; loss of Agr QS activity would phenotypically resemble a S. aureus commensal state that would be unlikely to be associated with disease. Commensal competition resulting in dampened virulence of the competitor may represent an exciting and unexplored possibility for development of novel antimicrobial compounds.IMPORTANCE While some individuals are nasally colonized with S. aureus, the underlying factors that determine colonization are not understood. There is increasing evidence that indicates that resident bacteria play a role; some commensal species can eradicate S. aureus from the nasal cavity. Among these, Corynebacterium pseudodiphtheriticum can eliminate S. aureus from the human nose. We sought to understand this phenomenon at a molecular level and found that C. pseudodiphtheriticum produces a factor(s) that specifically kills S. aureus While resistant S. aureus isolates were recovered at a low frequency, resistance came at the cost of attenuated virulence in these strains. Molecular dissection of the specific strategies used by C. pseudodiphtheriticum to kill S. aureus could lead to the development of novel treatments or therapies. Furthermore, commensal competition that requires virulence components of the competitor may represent an exciting and unexplored possibility for development of novel antimicrobial compounds.

Sequential assembly of the septal cell envelope prior to V snapping in Corynebacterium glutamicum.

Members of the Corynebacterineae, including Corynebacterium and Mycobacterium, have an atypical cell envelope characterized by an additional mycomembrane outside of the peptidoglycan layer. How this multilayered cell envelope is assembled remains unclear. Here, we tracked the assembly dynamics of different envelope layers in Corynebacterium glutamicum and Mycobacterium smegmatis by using metabolic labeling and found that the septal cell envelope is assembled sequentially in both species. Additionally, we demonstrate that in C. glutamicum, the peripheral peptidoglycan layer at the septal junction remains contiguous throughout septation, forming a diffusion barrier for the fluid mycomembrane. This diffusion barrier is resolved through perforations in the peripheral peptidoglycan, thus leading to the confluency of the mycomembrane before daughter cell separation (V snapping). Furthermore, the same junctional peptidoglycan also serves as a mechanical link holding the daughter cells together and undergoes mechanical fracture during V snapping. Finally, we show that normal V snapping in C. glutamicum depends on complete assembly of the septal cell envelope.

Microbes Culture System Using Cellulose Tubes and Photocatalyst-Coated Glass Balls.

Currently, microbes are utilized in many fields, such as medicine, food and environment etc. For more application of microbes, we need a new culture system, which can culture target microbes in large quantities at low cost. Thereupon, we propose a culture system using cellulose tubes. Target microbes are encapsulated inside the cellulose tubes, where they acquire nutrients and oxygen through nano pores of the tubes and are protected from competitive microbes even in open environment. To further increase the amount of oxygen and nutritions available for the target microbes, we propose photocatalyst-coated glass balls (PCGB) to sterilize competing microbes outside the tubes. We experimentally verified the effectiveness of the proposed culture system by culturing Coryne gultamicum as the target microbes.

Visions of Eye Commensals: The Known and the Unknown About How the Microbiome Affects Eye Disease.

Until recently, the ocular surface is thought by many to be sterile and devoid of living microbes. It is now becoming clear that this may not be the case. Recent and sophisticated PCR analyses have shown that microbial DNA-based "signatures" are present within various ethnic, geographic, and contact lens wearing communities. Furthermore, using a mouse model of ocular surface disease, we have shown that the microbe, Corynebacterium mastitidis (C. mast), can stably colonize the ocular mucosa and that a causal relationship exists between ocular C. mast colonization and beneficial local immunity. While this constitutes proof-of-concept that a bona fide ocular microbiome that tunes immunity can exist at the ocular surface, there remain numerous unanswered questions to be addressed before microbiome-modulating therapies may be successfully developed. Here, the authors will briefly outline what is currently known about the local ocular microbiome as well as microbiomes associated with other sites, and how those sites may play a role in ocular surface immunity. Understanding how commensal microbes affect the ocular surface immune homeostasis has the potential revolutionize how we think about treating ocular surface disease.

A New Approach of Rpf Addition to Explore Bacterial Consortium for Enhanced Phenol Degradation Under High Salinity Conditions.

Only a small fraction of salt-tolerant phenol-degrading bacteria can be isolated by conventional plate separation methods, because most bacteria in nature are in a viable but non-culturable (VBNC) state. The aims of this study were to screen out more effective functional bacteria using resuscitation-promoting factor (Rpf), and to determine whether a mixed bacterial consortium possesses better phenol-degrading capabilities under high salinity conditions. The results indicated that three strains unique to treatment group with Rpf addition were obtained. A mixed bacterial consortium consisting of two high-efficient strains which belonged to genera Bacillus and Corynebacterium was capable of utilizing phenol as a sole source of carbon at high salinity. Complete degradation of 100 mg/L phenol at 2% NaCl concentration was achieved within 8 h. This study provides new insights into resuscitation of VBNC bacteria for enhanced treatment of phenol-laden saline wastewater.

New lipopeptide produced by Corynebacterium aquaticum from a low-cost substrate.

Conventional biosurfactants have high production costs. Therefore, the use of low-cost carbon sources for their production is attractive for industry. The ability to remain stable under various environmental conditions further extends industrial application. Here we aimed to evaluate the stability of a new lipopeptide produced by Corynebacterium aquaticum using fish residue as an unconventional energy source. The biosurfactant was produced using 3% fish residue, 2% of the microorganism, and mineral medium. Biosurfactant characterization was performed by thin layer chromatography (TLC), as well as by testing its infrared, surface tension, emulsifying activity, and ionic character. The stability of the biosurfactant was evaluated by testing its surface tension at a range of temperatures, pH, and saline concentrations, as well as after 6 months of storage. The biosurfactant was characterized as a lipopeptide due to its retention time, which was coincident with the amino acid and lipid chains obtained in the TLC analysis, being confirmed by some regions of absorption verified in the infrared analysis. The surface tension and emulsifying activity of the biosurfactant were 27.8 mN/m and 87.6%, respectively, and showed anionic character. The biosurfactant was stable at temperatures of 20 to 121 °C, in saline concentrations of 1 to 7%, and at pH close to neutrality. Based on our findings, it is possible to use unconventional sources of energy to produce a lipopeptide biosurfactant that can act under various environments.

Novel selective medium for the isolation of corynebacterium kroppenstedtii from heavily colonised clinical specimens.

AIMS: Granulomatous mastitis due to Corynebacterium kroppenstedtii is an increasingly recognised cause of an indolent and distressing mastitis in non-lactating females. This slow-growing lipophilic organism is not reliably isolated using routine culture methods. A novel selective culture medium (CKSM) is designed to optimise the isolation of this organism from clinical specimens. METHODS: CKSM contains 10% galactose and Tween 80 (10%) to enhance the growth of C. kroppenstedtii, fosfomycin (100 µg/mL) to suppress the other bacteria, and differentiate C. kroppenstedtii from non-kroppenstedtii lipophilic corynebacteria by esculin hydrolysis. The medium was evaluated for its ability to support the growth of C. kroppenstedtii, selection and differentiation of C. kroppenstedtii from other bacteria in non-sterile clinical specimens. RESULTS: C. kroppenstedtii grew as 1-2 mm colonies with black halo on CKSM within 72 hours of incubation, compared with barely visible pinpoint colonies on routine blood agars. During the four-month period of evaluation with 8896 respiratory specimens, 103 breast specimens, 1903 female genital tract specimens, 617 newborn surface swabs and 10 011 miscellaneous specimens, 186 C. kroppenstedtii were isolated, including 127 (1.4%) respiratory and 59 (0.5%) miscellaneous specimens, 184 of them were found only on CKSM. Besides the three (2.9%) positive breast specimens, 27 (1.4%) high vaginal and endocervical swabs, and 11 (1.8%) surface swabs of newborns were positive for C. kroppenstedtii. CONCLUSIONS: CKSM is a useful addition to routine agar media for the isolation of C. kroppenstedtii, and will be helpful for studying the epidemiology and transmission of this unusual Corynebacterium causing granulomatous mastitis.

Characterization of dioxygenases and biosurfactants produced by crude oil degrading soil bacteria

ABSTRACT Role of microbes in bioremediation of oil spills has become inevitable owing to their eco friendly nature. This study focused on the isolation and characterization of bacterial strains with superior oil degrading potential from crude-oil contaminated soil. Three such bacterial strains were selected and subsequently identified by 16S rRNA gene sequence analysis as Corynebacterium aurimucosum, Acinetobacter baumannii and Microbacterium hydrocarbonoxydans respectively. The specific activity of catechol 1,2 dioxygenase (C12O) and catechol 2,3 dioxygenase (C23O) was determined in these three strains wherein the activity of C12O was more than that of C23O. Among the three strains, Microbacterium hydrocarbonoxydans exhibited superior crude oil degrading ability as evidenced by its superior growth rate in crude oil enriched medium and enhanced activity of dioxygenases. Also degradation of total petroleum hydrocarbon (TPH) in crude oil was higher with Microbacterium hydrocarbonoxydans. The three strains also produced biosurfactants of glycolipid nature as indicated d by biochemical, FTIR and GCMS analysis. These findings emphasize that such bacterial strains with superior oil degrading capacity may find their potential application in bioremediation of oil spills and conservation of marine and soil ecosystem.

Characterization of dioxygenases and biosurfactants produced by crude oil degrading soil bacteria.

Role of microbes in bioremediation of oil spills has become inevitable owing to their eco friendly nature. This study focused on the isolation and characterization of bacterial strains with superior oil degrading potential from crude-oil contaminated soil. Three such bacterial strains were selected and subsequently identified by 16S rRNA gene sequence analysis as Corynebacterium aurimucosum, Acinetobacter baumannii and Microbacterium hydrocarbonoxydans respectively. The specific activity of catechol 1,2 dioxygenase (C12O) and catechol 2,3 dioxygenase (C23O) was determined in these three strains wherein the activity of C12O was more than that of C23O. Among the three strains, Microbacterium hydrocarbonoxydans exhibited superior crude oil degrading ability as evidenced by its superior growth rate in crude oil enriched medium and enhanced activity of dioxygenases. Also degradation of total petroleum hydrocarbon (TPH) in crude oil was higher with Microbacterium hydrocarbonoxydans. The three strains also produced biosurfactants of glycolipid nature as indicated d by biochemical, FTIR and GCMS analysis. These findings emphasize that such bacterial strains with superior oil degrading capacity may find their potential application in bioremediation of oil spills and conservation of marine and soil ecosystem.

Beneficial and Safety Properties of a Corynebacterium vitaeruminis Strain Isolated from the Cow Rumen.

Corynebacterium vitaeruminis MRU4 was isolated from the cow rumen and was differentiated from other isolates by rep-PCR and RAPD and identified by 16S rRNA sequencing. This strain presented higher survival rates for low pH and bile salts treatments, and it was able to survive and multiply in simulated gastric and intestinal environments. C. vitaeruminis MRU4 had a 53.2% auto-aggregation rate, 42.4% co-aggregation rate with Listeria monocytogenes Scott A, 41.6% co-aggregation rate with Enterococcus faecalis ATCC 19443, 10.0% co-aggregation rate with Lactobacillus sakei ATCC 15521, and 98.2% cell surface hydrophobicity rate. PCR analysis showed the presence of EFTu and map genes. The strain possessed positive results for deconjugation of bile salts (taurocholic acid, taurodeoxycholic acid, glycocholic acid, and glycodeoxycholic acid) and positive results for ß-galactosidase activity and lactose assimilation activity (glucose of 8.15 ± 0.01 CFU/ml and lactose of 9.24 ± 0.02 CFU/ml). No virulence was observed by phenotypical tests. C. vitaeruminis MRU4 was resistant to oxacillin, gentamicin, erythromycin, clindamycin, sulfa/trimethoprim, and rifampicin by the disc diffusion method and showed resistance just for vancomycin by the Etest® strips test. The strain was negative for 50 tested virulence and resistance genes based on performed PCR. Based on our knowledge, this is the first report regarding the beneficial potential of one C. vitaeruminis strain.

Development of a systematic feedback isolation approach for targeted strains from mixed culture systems.

Elucidation of functions of bacteria in a mixed culture system (MCS) such as composting, activated sludge system is difficult, since the system is complicating with many unisolated bacteria. Here, we developed a systematic feedback isolation strategy for the isolation and rapid screening of multiple targeted strains from MCS. Six major strains (Corynebacterium sphenisci, Bacillus thermocloacae, Bacillus thermoamylovorans, Bacillus smithii, Bacillus humi, and Bacillus coagulans), which are detected by denaturing gradient gel electrophoresis (DGGE) analysis in our previous study on MCS for l-lactic acid production, were targeted for isolation. Based on information of suitable cultivation conditions (e.g., media, pH, temperature) from the literature, feedback isolation was performed to form 136 colonies. The following direct colony matrix assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) was optimised as the second screening to narrow down 20 candidate colonies from similar spectra patterns with six closest type strains. This step could distinguish bacteria at the species level with distance similarity scores ≥0.55 corresponding to 16S rRNA gene sequence similarity ≥98.2%, suggesting that this is an effective technique to minimize isolates close to targeted type strains. Analysis of 16S rRNA gene sequences indicated that two targeted strains and one strain related to the target had successfully been isolated, showing high similarities (99.5-100%) with the sequences from the DGGE bands, and that the other candidates were affiliated with three strains that were closely related to the target species. This study proposes a new method for systematic feedback isolation that may be useful for isolating targeted strains from MCS for further investigation.

[ANTIBIOTICS RESISTANCE OF CORYNEBACTERIUM NON DIPHTHERIAE STRAINS].

AIM: Study the frequency of occurrence of antibiotics resistant strains of various species of Corynebacterium non diptheriae. MATERIALS AND METHODS: C.pseudodiphtheriticum, C.pseudo- tuberculosis, C.xerosis, C.amycolatum, C.striatum, C. ulcerans strains isolated from patients with pathologies of respiratory and urogenital tract, as well as individuals taking prophylaxis ex- amination were used. Sensitivity to antibacterial preparations was determined by the serial dilution method. RESULTS: The highest number of Coryniebacterium non diptheriae strains dis- played resistance to benzylpenicillin (54.8%) and lincomycin (50.7%), and lowest - to cefotaxime, cefazolin (6.8%) and vancomycin (13.7%). The highest number of antibiotics resistant strains were detected among members of C.pseudotuberculosis (100%), C.xerosis (96.0%) and C. pseudodiphtheriticum (81.0%) species. Polyresistant strains were detected most frequently among C.xerosis, C.amycolatum and C.striatum species. Strains of Corynebacterium non diptheriae most frequently displayed resistance to 1 or 2 antibacterial preparations (24.7%), less frequently - to 3 (20.5%); 4 (13.7%), 5 (4.1%) and 6 (1.4%) preparations. CONCLUSION: The amount of antibiotics resistant strains of Cdrynebacterium non diptheriae is large (89.0%) and non-similar in various species.