METHYLATED AMINE-UTILISING BACTERIA AND MICROBIAL NITROGEN CYCLING IN MOVILE CAVE

Movile Cave is an unusual, isolated ecosystem which harbours a complex population of microorganisms, fungi and endemic invertebrates. In the absence of light and with no fixed carbon entering the cave, life is sustained by non-phototrophic microorganisms such as sulfur and methane oxidisers. Also present are methylotrophs that use one-carbon compounds such as methanol and methylated amines as their sole source of carbon and energy. Produced during putrefaction, methylated amines are likely to be major degradation products in Movile Cave. Further to being methylotrophic substrates, they are also a nitrogen source for many non-methylotrophic bacteria. The role of methylated amines as carbon and nitrogen sources for Movile Cave bacteria was investigated using a combination of DNA stable isotope probing and cultivation studies. Both, well-characterised and novel methylotrophs were identified: Methylotenera mobilis dominated 13C-monomethylamine SIP enrichments, while members of Catellibacterium, Cupriavidus and Altererythrobacter were also active. Cultivation studies consolidated SIP results in obtaining the first methylotrophic isolates from the genera Catellibacterium and Mesorhizobium. Pathways for monomethylamine (MMA) metabolism were investigated using new PCR primers designed to target gmaS, the gene for gammaglutamylmethylamide synthetase, a key enzyme of the recently characterised indirect MMA oxidation pathway. This pathway is also present in bacteria that use MMA only as a nitrogen source, while the well-characterised, direct MMA oxidation pathway involving methylamine dehydrogenase (mauA) is found only in methylotrophs. gmaS was present in all MMAutilising isolates, while mauA was found only in some methylotrophs, suggesting the indirect pathway is the major mode of MMA oxidation both in methylotrophs and non-methylotrophs from Movile Cave. Preliminary gmaS surveys revealed a high diversity of gmaS-containing bacteria. The roles of N2 fixers and nitrifiers were also investigated. Both bacterial and archaeal ammonia oxidisers were found to be active; however, sulfur oxidisers appeared to be the dominant autotrophs in Movile Cave

Draft Genome Sequences of Facultative Methylotrophs, Gemmobacter sp. Strain LW1 and Mesorhizobium sp. Strain 1M-11, Isolated from Movile Cave, Romania

Facultative methylotrophs belonging to the genera Gemmobacter and Mesorhizobium were isolated from microbial mat and cave water samples obtained from the Movile Cave ecosystem. Both bacteria can utilize methylated amines as their sole carbon and nitrogen source. Here, we report the draft genome sequences of Gemmobacter sp. strain LW1 and Mesorhizobium sp. strain IM1

Novel Antimicrobial Cellulose Fleece Inhibits Growth of Human-Derived Biofilm-Forming Staphylococci During the SIRIUS19 Simulated Space Mission

Two novel antimicrobial surface coatings were assessed for their lasting antibacterial effect under simulated space conditions during the SIRIUS-19 study. Because long-term space travel can affect the human immune system, astronauts are particularly susceptible to infectious disease. Moreover, the space flight environment can alter the composition of microbial communities within the spacecraft and increase bacterial virulence and resistance to antibiotics. In addition to protecting the crew from infection by human pathogens, prevention and elimination of bacterial contamination is important to avoid corrosion and damage of the technical equipment. The antimicrobial coating AGXX(R)consists of micro-galvanic cells composed of silver and ruthenium which damage bacterial cells through the release of reactive oxygen species. Over the last years, several studies on the antimicrobial effect of AGXX(R)have demonstrated an effective inhibition of growth and even complete elimination of many pathogenic bacteria - including multiresistant microorganisms - as well as their biofilms. The second antimicrobial coating, GOX, consists of chemically modified graphene oxide. Through a positive surface charge and its flexible scaffold, GOX can multivalently bind and immobilize bacteria via electrostatic attraction. Here, AGXX(R)and GOX were applied to non-metallic carriers not previously tested. The antimicrobial coated materials, as well as uncoated control samples, were exposed in the SIRIUS artificial space module and analyzed at different time points during the 4-months isolation study. Survival and growth of airborne heterotrophic, aerobic bacteria on the surfaces were assessed by cultivation-based methods, employing growth conditions suitable for potential human pathogens. Human-associated, biofilm-forming Staphylococcus spp. (S. hominis, S. haemolyticus, and S. epidermidis) strongly dominated at all time points, most were resistant against erythromycin, kanamycin, and ampicillin. AGXX(R)coatings completely inhibited growth of these opportunistic pathogens on all tested surface materials. Particularly, AGXX(R)-cellulose fleece achieved a clear reduction in bacterial load able to recover post contact. GOX-cellulose fleece effectively immobilized bacteria. Sequence analysis of 16S rRNA gene amplicons revealed that the isolated Staphylococcus spp. did not dominate the overall bacterial community, accounting for only 0.1-0.4% of all sequences. Instead, molecular data revealed Lactobacillus, Comamonas, Pseudomonas, Sporosarcina, and Bacillusas the dominant genera across all samples and time points

Comparative genomics analyses indicate differential methylated amine utilization trait within members of the genus Gemmobacter

Methylated amines are ubiquitous in the environment and play a role in regulating the earth's climate via a set of complex biological and chemical reactions. Microbial degradation of these compounds is thought to be a major sink. Recently we isolated a facultative methylotroph, Gemmobacter sp. LW‐1, an isolate from the unique environment Movile Cave, Romania, which is capable of methylated amine utilization as a carbon source. Here, using a comparative genomics approach, we investigate how widespread methylated amine utilization is within members of the bacterial genus Gemmobacter. Seven genomes of different Gemmobacter species isolated from diverse environments, such as activated sludge, fresh water, sulphuric cave waters (Movile Cave) and the marine environment were available from the public repositories and used for the analysis. Our results indicate that methylamine utilization is a distinctive feature of selected members of the genus Gemmobacter, namely G. aquatilis, G. lutimaris, G. sp. HYN0069, G. caeni and G. sp. LW‐1 have the genetic potential while others (G. megaterium and G. nectariphilus) have not

К проблеме социальной эффективности инноваций в профессиональном образовании

Movile Cave, Romania, is an unusual underground ecosystem that has been sealed off from the outside world for several million years and is sustained by non-phototrophic carbon fixation. Methane and sulfur-oxidising bacteria are the main primary producers, supporting a complex food web that includes bacteria, fungi and cave-adapted invertebrates. A range of methylotrophic bacteria in Movile Cave grow on one-carbon compounds including methylated amines, which are produced via decomposition of organic-rich microbial mats. The role of methylated amines as a carbon and nitrogen source for bacteria in Movile Cave was investigated using a combination of cultivation studies and DNA stable isotope probing (DNA-SIP) using 13C-monomethylamine (MMA). Two newly developed primer sets targeting the gene for gamma-glutamylmethylamide synthetase (gmaS), the first enzyme of the recently-discovered indirect MMA-oxidation pathway, were applied in functional gene probing. SIP experiments revealed that the obligate methylotroph Methylotenera mobilis is one of the dominant MMA utilisers in the cave. DNA-SIP experiments also showed that a new facultative methylotroph isolated in this study, Catellibacterium sp. LW-1 is probably one of the most active MMA utilisers in Movile Cave. Methylated amines were also used as a nitrogen source by a wide range of non-methylotrophic bacteria in Movile Cave. PCR-based screening of bacterial isolates suggested that the indirect MMA-oxidation pathway involving GMA and N-methylglutamate is widespread among both methylotrophic and non-methylotrophic MMA utilisers from the cave

Microbiology of Movile Cave - A Chemolithoautotrophic Ecosystem

Discovered in 1986, Movile Cave is an unusual cave ecosystem sustained by in situ chemoautotrophic primary production. The cave is completely isolated from the surface and the primary energy sources are hydrogen sulfide and methane released from hydrothermal fluids. Both condensation and acid corrosion processes contribute to the formation of Movile Cave. Invertebrates, many of which are endemic to Movile Cave, are isotopically lighter in both carbon and nitrogen than surface organisms, indicating that they derive nutrition from chemoautotrophic primary producers within the cave. Here we review work on the microbiology of the Movile Cave ecosystem, with particular emphasis on the functional diversity of microbes involved in sulfur, carbon and nitrogen cycling, and discuss their role in chemosynthetic primary production