Non-spore Forming Research Articles

Isolation a new strain of Kocuria rosea capable of tolerating extreme conditions

A new actinobacterial strain was isolated from Ab-e-Siah spring (dark water) taken from the Ramsar city in Iran, and subjected to several stress conditions investigation. The isolate, named MG2 strain, was Gram-positive, aerobic, diplococci or tetrad shaped, non-spore forming and non-motile. Phylogenetic analysis of the isolate using 16S rDNA sequence indicated that the organism matched best with the genus Kocuria and the highest sequence similarities (98.55%) being found with Kocuria rosea. The 16S rDNA sequence determined in this study has been deposited in the NCBI database with the accession no. JX534199, K. rosea strain MG2. The isolated strain was an alkaliphilic-mesophilic bacterium because the optimal growth was observed at pH 9.2 and temperature of 28 °C under aerobic condition. MG2 was a halotolerant strain and tolerated maximally to 15% NaCl concentraion. Viability analysis by flow cytometry indicated that this strain had highly resistance to UV-C radiation and moderately resistance to desiccation after 28 days. The viability of K. rosea strains MG2 and Deinococcus radiodurans R1 were determined D87 and D98 according to D index, respectively, by a dose radiation 25 J/cm (Appukuttan et al., 2006). Thus the UV resistance of strain MG2 was comparable with representative radiation resistant Deinococcus. Also MG2 was grown at 1–4% of H2O2 as an oxidant agent. This research is the first study on multiple extreme resistance of Kocuria rosea new strain (MG2) isolated in Iran.

Effect of Radiofrequency Radiation from Telecommunication Base Stations on Microbial Diversity and Antibiotic Resistance

We aimed to investigate the effects of radiofrequency radiation from telecommunication on bacteria diversity and antibiotic sensitivity of surrounding bacteria micro-flora. In all cases of bacteria isolated, results indicated that radiation majorly influences and selected bacteria of specific morphology (rod-like, gram positive and spore formers) belonging to the genera Bacillus, Clostridium, Corynebacterium and Sporosarcina. Spore formation have been associated with prevention of DNA damage, this suggest that radiation may imposes stress detrimental to various non-spore forming, gram negative bacteria of nonrod morphology. The antibiotic resistance pattern indicated resistance above 50% in three of the ten common antibiotics tested with the highest of 96% recorded against zinnacef, followed by amoxicillin (70%) and ampiclox (56%). The highest resistance recorded was attributed to Bacillus spp while lowest resistance was recorded in Corynebacterium. The two largest groups of isolates; Bacillus and Clostridium have been reported as human pathogens and found to be multidrug resistant. © JASEM

Putting the Jigsaw Together - A Brief Insight Into the Tularemia

Abstract Tularemia is a debilitating febrile and potentially fatal zoonotic disease of humans and other vertebrates caused by the Gram-negative bacterium Francisella tularensis. The natural reservoirs are small rodents, hares, and possibly amoebas in water. The etiological agent, Francisella tularensis, is a non-spore forming, encapsulated, facultative intracellular bacterium, a member of the γ-Proteobacteria class of Gram-negative bacteria. Francisella tularensis is capable of invading and replicating within phagocytic as well as non-phagocytic cells and modulate inflammatory response. Infection by the pulmonary, dermal, or oral routes, respectively, results in pneumonic, ulceroglandular, or oropharyngeal tularemia. The highest mortality rates are associated with the pneumonic form of this disease. All members of Francisella tularensis species cause more or less severe disease Due to their abilities to be transmitted to humans via multiple routes and to be disseminated via biological aerosol that can cause the disease after inhalation of even an extremely low infectious dose, Francisella tularensis has been classified as a Category A bioterrorism agent. The current standard of care for tularemia is treatment with antibiotics, as this therapy is highly effective if used soon after infection, although it is not, however, absolutely effective in all cases.

Pedobacter bambusae sp. nov., isolated from soil of a bamboo plantation.

An ivory white, rod-shaped, non-motile, non-spore forming, Gram-staining-negative, and aerobic bacterium designated THG-G118T, was isolated from a soil sample collected in a bamboo plantation in Seoul, Republic of Korea. The 16S rRNA gene sequence analysis revealed a clear affiliation of strain THG-G118T with the genus Pedobacter. Strain THG-G118T was found to show the closest phylogenetic relationship with Pedobacter metabolipauper WB2.3-71T with 16S rRNA gene sequence similarity of 97.3 %. The major cellular fatty acids of strain THG-G118T were identified as C16:0, C18:0, iso-C15:0 and summed feature 3 (C16:1 ω7c and/or C16:1 ω6c), and the predominant menaquinone as menaquinone MK-7. The major polar lipids were found to be phosphatidylethanolamine (PE), unidentified phospholipid and unidentified aminophospholipids. Strain THG-G118T was found to contain ceramide phosphorylethanolamine 2 (CerPE-2) and dihydrosphingosine as the major ceramide. The G+C content of the genomic DNA was found to be 41.7 mol%. DNA-DNA hybridization experiments revealed a low level of DNA-DNA relatedness (<42.5 %) between strain THG-G118T and its closest relatives. The phenotypic, chemotaxonomic and phylogenetic data showed that strain THG-G118T could be clearly distinguished from the members of the genus Pedobacter. Based on the results of a polyphasic study, a new species, Pedobacter bambusae sp. nov., is proposed. The type strain is THG-G118T (=KACC 17544T = JCM 19364T).

Nioella nitratireducens gen. nov., sp. nov., a novel member of the family Rhodobacteraceae isolated from Azorean Island.

A novel Gram-negative, non-spore forming, rod-shaped aerobic bacterium, designated SSW136(T), was isolated from a surface seawater sample collected at Espalamaca (in Faial Island), Azores. Growth was found to occur from 10 to 37 °C, pH 6.0-8.0, and with 2-11 % of NaCl. 16S rRNA gene sequence indicated that the strain SSW136(T) belongs to the family Rhodobacteraceae. Strain SSW136(T) exhibited 96.3, 95.9, 95.7 and 95.5 sequence similarity to the type strains Oceanicola litoreus M-M22(T), Roseovarius aestuarii SMK-122(T), Marivita geojedonensis DPG-138(T), and Pseudoruegeria aquimaris SW-255(T) respectively. Neighbour-joining and maximum-parsimony phylogenetic trees based on 16S rRNA gene sequences revealed that strain SSW136(T) was affiliated to the family Rhodobacteraceae and formed a separate branch. The G+C content was 63.5 mol%. The major respiratory quinone was found to be Q-10. The polar lipids of strain SSW136(T) consisted of phosphatidylcholine, phosphatidylethanolamine, phosphatidylglycerol, two unidentified aminolipids and three unidentified phospholipids. The major fatty acids were C18:1 ω7c (46.5 %), Cyclo-C19:0 ω8c (16.0 %) and C16:0 (12.8 %). On the basis of the morphological, genotypic, chemotaxonomic characteristics and low DNA-DNA relatedness, strain SSW136(T) is proposed to represent a novel genus and novel species, Nioella nitratireducens gen. nov., sp. nov., in the family Rhodobacteraceae. The type strain is SSW136(T) (=KCTC 32417(T) = NCIM 5499(T)).

Mageeibacillus indolicus gen. nov., sp. nov.: A novel bacterium isolated from the female genital tract

Three isolates of a bacterium recovered from human endometrium using conventional culture methods were characterized biochemically and subjected to 16S rRNA gene sequencing and phylogenetic analysis. Isolates were non-motile, obligately anaerobic, non-spore forming, asaccharolytic, non-cellulolytic, indole positive, Gram positive rods. Cell wall fatty acid profiling revealed C14:0, C16:0, C18:2 ω6, 9c, C18:1 ω9c and C18:0 to be the major fatty acid composition. The DNA mol % G+C was determined to be 44.2%. 16S rRNA gene sequence analysis revealed only 91% sequence similarity with the closest cultivated bacterial isolate, Saccharofermentans acetigenes. Based on genotypic and phenotypic data, all three isolates are considered to be members of the same species and data suggest it represents a novel genus and species in the order Clostridiales with an association with Clostridium rRNA cluster III within the family Ruminococcaceae. We propose the name, Mageeibacillus indolicus gen. nov., sp. nov. The type strain is BAA-2120T and CCUG 59143T.

Deinococcus enclensis sp. nov., isolated from a marine sediment sample.

A novel pale-pink coloured strain, designated NIO-1023(T), was isolated from a marine sediment sample from Chorao Island, Goa, India. The taxonomic position of strain NIO-1023(T) was investigated by using a polyphasic approach. The cells were observed to be Gram-stain positive, coccal shaped and non-spore forming. Phylogenetic analyses using the 16S rRNA gene sequence of the isolate indicated that the organism belongs to the genus Deinococcus. The strain NIO-1023(T) showed highest 16S rRNA gene sequence similarities with Deinococcus ficus (97.8%), whereas other Deinococcus species showed less than 95% sequence similarity. The DNA-DNA relatedness with respect to D. ficus CC-FR2-10(T) was 23.9%. Chemotaxonomic data revealed that strain NIO-1023(T) contains only menaquinone MK-8 as the respiratory quinone and a complex polar lipid profile consisting of different unidentified glycolipids and polar lipids, two unknown phospholipids and three unknown phosphoglycolipids. As in other deinococci, one of these phosphoglycolipids was predominant in the profile. The predominant fatty acids were identified as C17:1 w8c, C16:1 w6c/w7c, C15:1 w6c and C17:1 w9c. The genomic DNA G+C content of strain NIO-1023(T) was determined to be 67.2mol%. The biochemical and chemotaxonomic properties demonstrate that strain NIO-1023(T) represents a novel species, for which the name Deinococcus enclensis sp. nov. is proposed. The type strain is NIO-1023(T) (=DSM 25127(T)=NCIM 5456(T)).

Flavobacterium arsenitoxidans sp. nov., an arsenite-oxidizing bacterium from Thai soil

An arsenite-oxidizing bacterium, strain S2-3H(T), was isolated from arsenic-contaminated soil sample collected from Dantchaeng district, Suphanburi province, Thailand and was characterized based on polyphasic taxonomic study. The strain was observed to be a Gram-stain negative, aerobic, yellow pigmented, non-spore forming and rod-shaped bacterium. Major menaquinone was MK-6. Iso-C15:0, iso-C15:0 3OH, C16:1 ω7c/C16:1 ω6c, C16:0, iso-C17:0 3OH, and C16:0 3OH were the predominant cellular fatty acids. The polar lipid profile consisted of phosphatidylethanolamine, unidentified phospholipids and unidentified aminophospholipids. The DNA G+C content was 37.0 mol%. Phylogenetic analysis using 16S rRNA sequence showed that strain S2-3H(T) is affiliated to the genus Flavobacterium, and is closely related to F. defluvii KCTC 12612(T) (97.0 %) and F. johnsoniae NBRC 14942(T) (97.0 %). The strain S2-3H(T) could be clearly distinguished from the related Flavobacterium species by its physiological and biochemical characteristics as well as its phylogenetic position and DNA-DNA relatedness. Therefore, the strain represents a novel species of the genus Flavobacterium, for which the name Flavobacterium arsenitoxidans sp. nov. (type strain S2-3H(T) = KCTC 22507(T) = NBRC 109607(T) = PCU 331(T) = TISTR 2238(T)) is proposed.

Molecular charcterization of tatD DNAse gene from Ralstonia paucula RA4T soil bacterium

Ralstonia paucula strain RA4T, a gram negative, non-spore forming, motile bacterium having positive catalase and oxidase test, was isolated from surface soil. Twin arginine translocation protein type D (TatD) is shown to be located in cytoplasm and exhibits magnesium-dependent DNase. A tatD DNase gene was isolated and cloned from Ralstonia paucula RA4T genome. Nucleotide sequence analysis of the gene revealed 813 nucleotides encoding a protein of 270 amino acid residues. The tatD gene showed a high similarity to homolog gene from Ralstonia pickettii strain 12D. The deduced polypeptide sequence of TatD DNase from R. paucula RA4T had a typical catalytic site, HHPLDEHRHDP, and its calculated molecular mass and predicted isoelectric point were 29616 Da and 5.33, respectively. The deduced amino acid sequence showed a high degree of similarity to TatD DNase isoforms from Ralstonia genus and other sources. Predicted three-dimensional structure of TatD confirmed the presence of active site and theoretical function as DNase.

Cloning, Expression and Characterisation of Recombinant Outer Membrane Protein 16 from Brucella spp.

Brucellosis is a disease caused by Brucella sp which is an aerobic, non-spore forming, non-motile gram negative coccobacillus. It is a National Institute of Health category B bioterror threat agent that causes incapacitating zoonosis. It is an emerging zoonosis, and an economically important infection of humans and livestock with a worldwide distribution. Medical defence against the bioterror threat posed by Brucella would be strengthened by development of a vaccine and improved diagnostic tests. Central to advancement of these goals is the discovery of bacterial constituents that are immunogenic or antigenic for humans and animals. Outer membrane proteins are particularly attractive for this purpose. In the present study, recombinant outer membrane protein 16 was expressed from Brucella abortus using prokaryotic expression vector and was characterized immunologically. The recombinant protein was purified and characterized immunologically by western blotting so that it can be used for development of enzyme linked immunosorbent assay or can be evaluated as vaccine candidate, in future.

Radiation resistant of native Deinococcus spp. isolated from the Lout desert of Iran “the hottest place on Earth”

Two native ionizing radiation-resistant bacteria were isolated and identified from a soil sample collected from extreme conditions of the Lout desert in Iran. The hottest land surface temperature has been recorded in the Lout desert from 2004 to 2009. Also, it is categorized as a hyper arid place. Both ionizing radiation and desiccation may cause damage on genome. Soil sample was irradiated in order to eliminate sensitive bacteria then cultured in one-tenth-strength tryptic soy broth medium. Bacterial suspension used for radiation treatment. Morphological and physiological characterization and phylogenetic studies based on 16S rRNA gene sequence were used for identification. The cells were rod shape, non-motile, non-spore forming and gram positive. The 16S rRNA gene sequence showed 99.5 % of similarity to Deinococcus ficus. Phylogenetic dendrogram demonstrated that the isolates branched with D. xibeiensis, D. ficus and D. mumbaiensis. Both isolates were resistant to >15 kGy of gamma radiation and >600 J m2 of UV radiation. This is the first report on radiation resistant bacteria belonging to genus Deinococcus isolated from the Lout desert of Iran.

First Report of Pantoea beijingensis Induced Soft Rot Disease of Pleurotus nebrodensis in China.

Pleurotus nebrodensis is a popular edible fungus in China. In December 2009, soft rot disease appeared on fruiting bodies of P. neberodensis in greenhouses located at Haidian District of Beijing, China. Early symptoms were water-soaked lesions on the pilei of the fruiting bodies. Lesions then spread and purulent tissues were formed. Severe soft rot induced production of deformed fruiting bodies and offensive odor. Diseased fruiting bodies stopped growth and commercial losses were estimated at 5 to 10%. Internal sections of the diseased pilei about 0.5 × 0.5 cm were suspended in 0.85% NaCl and the suspension was spread on trypticase soy agar (TSA) media. After incubation at 30°C for 2 days, dominant bacterial colonies were yellow, smooth, round, and convex. Cells were gram-negative, short rods, non-capsulated, motile, and non-spore forming. The 16S rRNA gene (1,408 bp, GenBank Accession No. KF849293) was amplified by using the universal forward primer P1 (5'-AGAGTTTGATCCTGGTCAGAACGCT-3') and the universal reverse primer P6 (5'-TACGGCTACCTTGTTACGACTTCACCCC-3'). Neighbor-joining tree analysis based on 16S rRNA gene sequences demonstrated the isolates belonged to the genus Pantoea and showed highest similarity with Pantoea beijingensis LMG 27579T (99.9% similarity, 2 base pair differences), which was isolated from the diseased fruiting bodies of P. eryngii in China. Pantoea sp. PA4, reported to be the pathogen of soft rot disease of P. eryngii in Korea (1), showed 99.8% similarity with P. ananatis ATCC 33244T, which suggested our isolates belonged to a different species with Pantoea sp. PA4 (supported by 97.9% similarity between the present isolates and Pantoea sp. PA4) (2). Results of physiological properties and enzyme activities determined by the API 20E, API 20NE, API 50CH, API ZYM (bioMérieux), and GN2 MicroPlate (Biolog) system showed that there was no difference between the present isolates and P. beijingensis LMG 27579T. Pathogenicity tests for these isolates were performed with bacterial suspensions (approximately 1 × 106 CFU/ml) that were grown for 24 h in trypticase soy broth (TSB). Fruiting bodies of P. nebrodensis were induced after spawn was run for 30 days in plastic bags. When the young fruiting bodies were forming, the prepared bacterial suspension was sprayed onto the surface of the pilei. Sterilized TSB media was used as a negative control. All inoculated fruiting bodies were then incubated at 16°C with 80 to 95% relative humidity. Assays were conducted twice and five fruiting bodies were used each time. Results were observed after 5 to 10 days of incubation. The symptoms that developed were similar to those observed in the original samples. The negative control remained symptomless. Koch's postulates were fulfilled by re-isolating bacteria, which were identical to the original isolates based on 16S rRNA gene sequence analysis, physiological properties and enzyme activities. Pantoea ananatis was first reported as a pathogen of Pleurotus eryngii, causing soft rot disease (1), but to the best of our knowledge, this is the first report of P. beijingensis- induced soft rot disease of P. nebrodensis in China. This disease may be an emerging disease problem in the near future and information on this pathogen will be useful in the development of management practices.

Isolation and characterization of deleterious Pseudomonas aeruginosa KC1 from rhizospheric soils and its interaction with weed seedlings

The bacterial isolate KC1 was screened from the rhizosphere of castor plants (Ricinus communis) indigenous to agricultural fields of Bihar. The isolate was Gram negative, non-spore forming, and exhibited fluorescence under UV light. Its molecular characterization is based on the sequencing of 16S rDNA (1450bp) and alignment at GeneBank (NCBI, MaryLand). The strain has been validated as Pseudomonas aeruginosa (HM195190). The bacterium grew at 4–42°C, with a temperature optima of 30°C. The strain KC1 was found to produce cyanide (4.78nmoll−1) over a period of 36h. Data revealed enhanced cyanogenesis (6.98nmoll−1), when glycine was provided in the King’s B medium. Seed bacterization exhibited reduction in root length, shoot length of weed seedlings (Amaranthus spinosus, Portulaca oleracea), which was significant (p<0.05) in both laboratory and glasshouse experiments. Biomass was significantly reduced (p<0.05) for the weed seedlings in glasshouse experiments. However, KC1 inoculated crop seedlings (Triticum aestivum) were found to be less inhibitory as compared to weed seedlings. The observations are significant to establish, that the secondary metabolites producing KC1 rhizobacterium, P. aeruginosa KC1 could be exploited as a weed biocontrol agent. The innate potential of KC1 could be further formulated and utilized in field applications for agricultural sustainability.

Typing of Streptococcus Milleri group by molecular and biochemical methods

Background: Streptococci detected in various regions of the body as normal flora are Gram positive diplococci, non-spore forming and non-motile bacteria. Streptococci can be classified according to biochemical and antigenic properties and the ability to produce hemolysis on blood agar. The diameter of beta hemolitic streptococci colonies are generally greater than 0.5mm and these are called pyogen; on the other hand bacteria whose colonies are smaller than 0.5mm (pin-point) are called Streptococcus milleri group in Europe and Streptococcus anginosus group in America. Streptococcus milleri group are classified into three species; Streptococcus anginosus, Streptococcus intermedius, Streptococcus constellatus. S. milleri group bacteria are genetically related to non-hemolitic streptococci. Methods & Materials: 100 streptococci strains were isolated from throat samples and also from other body sites such as abcess, blood and pustules. VP, CAMP tests, esculin and urease hydrolisis, sugar fermatation tests and Niven’s arginin hydrolysis test were carried out for pin-point colonies and species identification was carried by API 20 Strep kit and by PCR (table). Antibiotic susceptibility testing was carried out according to Clinical and Laboratory Standards Institute (CLSI) criteria. Results: All isolates were VP positive, PYR negative and did not hydrolyse hippurat; two strains were urease positive. All strains was found to be susceptible to vancomycin, ceftriaxone, cefepime, cefotaxime, levofloxacin, linezolid antibiotics. 16% of strains (10 S.anginosus, 4 S.constellatus)were found to be resistant to erythromycin. 6% of the strains (5 S.anginosus, 3 S.constellatus) were resistant to clindamycin and 5% (3 S.constellatusve 2 S.anginosus) of the strains were found to be resistant to tetracycline. According to PCR results of 100 isolates, 56 were identified as S.anginosus and 8 were identified as S.intermedius. Conclusion: It was determined that biochemical identification scheme is not satisfactory forStreptococcus milleri group. No emergence antibiotic resistance was determined. However, MIC should be investigated for strains isolated from sterile body parts. Commercially available biochemical kits are inadequent and identification should besupported with molecular methods. This study is thought to be a backbone for other studies in the future.

Early diagnosis of enteric fever in resource limited countries

Background: Streptococci detected in various regions of the body as normal flora are Gram positive diplococci, non-spore forming and non-motile bacteria. Streptococci can be classified according to biochemical and antigenic properties and the ability to produce hemolysis on blood agar. The diameter of beta hemolitic streptococci colonies are generally greater than 0.5mm and these are called pyogen; on the other hand bacteria whose colonies are smaller than 0.5mm (pin-point) are called Streptococcus milleri group in Europe and Streptococcus anginosus group in America. Streptococcus milleri group are classified into three species; Streptococcus anginosus, Streptococcus intermedius, Streptococcus constellatus. S. milleri group bacteria are genetically related to non-hemolitic streptococci. Methods & Materials: 100 streptococci strains were isolated from throat samples and also from other body sites such as abcess, blood and pustules. VP, CAMP tests, esculin and urease hydrolisis, sugar fermatation tests and Niven’s arginin hydrolysis test were carried out for pin-point colonies and species identification was carried by API 20 Strep kit and by PCR (table). Antibiotic susceptibility testing was carried out according to Clinical and Laboratory Standards Institute (CLSI) criteria. Results: All isolates were VP positive, PYR negative and did not hydrolyse hippurat; two strains were urease positive. All strains was found to be susceptible to vancomycin, ceftriaxone, cefepime, cefotaxime, levofloxacin, linezolid antibiotics. 16% of strains (10 S.anginosus, 4 S.constellatus)were found to be resistant to erythromycin. 6% of the strains (5 S.anginosus, 3 S.constellatus) were resistant to clindamycin and 5% (3 S.constellatusve 2 S.anginosus) of the strains were found to be resistant to tetracycline. According to PCR results of 100 isolates, 56 were identified as S.anginosus and 8 were identified as S.intermedius. Conclusion: It was determined that biochemical identification scheme is not satisfactory forStreptococcus milleri group. No emergence antibiotic resistance was determined. However, MIC should be investigated for strains isolated from sterile body parts. Commercially available biochemical kits are inadequent and identification should besupported with molecular methods. This study is thought to be a backbone for other studies in the future.

Typing of Streptococcus milleri group by molecular and biochemical methods

Background: Streptococci detected in various regions of the body as normal flora are Gram positive diplococci, non-spore forming and non-motile bacteria. Streptococci can be classified according to biochemical and antigenic properties and the ability to produce hemolysis on blood agar. The diameter of beta hemolitic streptococci colonies are generally greater than 0.5mm and these are called pyogen; on the other hand bacteria whose colonies are smaller than 0.5mm (pin-point) are called Streptococcus milleri group in Europe and Streptococcus anginosus group in America. Streptococcus milleri group are classified into three species; Streptococcus anginosus, Streptococcus intermedius, Streptococcus constellatus. S. milleri group bacteria are genetically related to non-hemolitic streptococci. Methods & Materials: 100 streptococci strains were isolated from throat samples and also from other body sites such as abcess, blood and pustules. VP, CAMP tests, esculin and urease hydrolisis, sugar fermatation tests and Niven’s arginin hydrolysis test were carried out for pin-point colonies and species identification was carried by API 20 Strep kit and by PCR (table). Antibiotic susceptibility testing was carried out according to Clinical and Laboratory Standards Institute (CLSI) criteria. Results: All isolates were VP positive, PYR negative and did not hydrolyse hippurat; two strains were urease positive. All strains was found to be susceptible to vancomycin, ceftriaxone, cefepime, cefotaxime, levofloxacin, linezolid antibiotics. 16% of strains (10 S.anginosus, 4 S.constellatus)were found to be resistant to erythromycin. 6% of the strains (5 S.anginosus, 3 S.constellatus) were resistant to clindamycin and 5% (3 S.constellatusve 2 S.anginosus) of the strains were found to be resistant to tetracycline. According to PCR results of 100 isolates, 56 were identified as S.anginosus and 8 were identified as S.intermedius. Conclusion: It was determined that biochemical identification scheme is not satisfactory forStreptococcus milleri group. No emergence antibiotic resistance was determined. However, MIC should be investigated for strains isolated from sterile body parts. Commercially available biochemical kits are inadequent and identification should besupported with molecular methods. This study is thought to be a backbone for other studies in the future.

Complete Genome sequence of Burkholderia phymatum STM815(T), a broad host range and efficient nitrogen-fixing symbiont of Mimosa species.

Burkholderia phymatum is a soil bacterium able to develop a nitrogen-fixing symbiosis with species of the legume genus Mimosa, and is frequently found associated specifically with Mimosa pudica. The type strain of the species, STM 815T, was isolated from a root nodule in French Guiana in 2000. The strain is an aerobic, motile, non-spore forming, Gram-negative rod, and is a highly competitive strain for nodulation compared to other Mimosa symbionts, as it also nodulates a broad range of other legume genera and species. The 8,676,562 bp genome is composed of two chromosomes (3,479,187 and 2,697,374 bp), a megaplasmid (1,904,893 bp) and a plasmid hosting the symbiotic functions (595,108 bp).

Comamonas faecalis sp. nov., Isolated from Domestic Pig Feces

A new bacterial strain, designated as FF42(T), was isolated from feces of domestic pigs-collected from Suwon, Korea-and was characterized to determine its taxonomic position. Strain FF42(T) was observed to be Gram negative, aerobic, non-spore forming, motile, and rod-shaped cells. Based on the phylogenetic and 16S rRNA sequence analyses, it was revealed that strain FF42(T) belonged to the genus Comamonas. The highest degree of sequence similarities was determined to be with Comamonas zonglianii BF-3(T) (96.3 %), Comamonas composti CC-YY287(T) (96.1 %), and Comamonas nitrativorans 23310(T) (95.9 %), while showing less than 95.6 % identity with the remaining Comamonas species. Growth of strain FF42(T) occurred between 25 and 40 °C (optimum, 28 °C) and at pH of 5-9 (optimum, pH 6.0). It grew in the presence of 0-3 % (w/v) NaCl while minimally tolerating at 3 % (w/v) NaCl. Biochemical and physiological tests revealed phenotypic differentiation of strain FF42(T) to other members of the genus Comamonas. The predominant quinone is ubiquinone (Q-8). The major cellular fatty acids were C10:0 3OH, C16:0, summed feature 3 (C16:1 ω7c/C16:1 ω6c), and summed feature 8 (C18:1 ω6c/C18:1 ω7c), all of which have previously been reported to occur in the species of the genus Comamonas. The G+C molar content for strain FF42(T) is 60.2 mol %. Based on phylogenetic and phenotypic analyses, strain FF42(T) (=KEMC 1002-058(T)=JCM 17561(T)) is clearly referred to be a novel species for the genus Comamonas, for which the name Comamonas faecalis sp. nov. is proposed.

Clostridium geopurificans Strain MJ1 sp. nov., A Strictly Anaerobic Bacterium that Grows via Fermentation and Reduces the Cyclic Nitramine Explosive Hexahydro-1,3,5-Trinitro-1,3,5-Triazine (RDX)

A fermentative, non-spore forming, motile, rod-shaped bacterium, designated strain MJ1(T), was isolated from an RDX contaminated aquifer at a live-fire training site in Northwest NJ, United States. On the basis of 16S rRNA gene sequencing and DNA base composition, strain MJ1(T) was assigned to the Firmicutes. The DNA G+C content was 42.8 mol%. Fermentative growth was supported by glucose and citrate in a defined basal medium. The bacterium is a strict anaerobe that grows between at pH 6.0 and pH 8.0 and 18 and 37 °C. The culture did not grow with hexahydro-1,3,5-trinitro-1,3,5-triazine (RDX) as the electron acceptor or mineralize RDX under these conditions. However, MJ1(T) transformed RDX into MNX, methylenedinitramine, formaldehyde, formate, ammonium, nitrous oxide, and nitrate. The nearest phylogenetic relative with a validly published name was Desulfotomaculum guttoideum (95 % similarity). However, MJ1(T) was also related to Clostridium celerecrescens DSM 5628 (95 %), Clostridium indolis DSM 755 (94 %), and Clostridium sphenoides DSM 632 (94 %). DNA:DNA hybridization with these strains was between 6.7 and 58.7 percent. The dominant cellular fatty acids (greater than 5 % of the total, which was 99.0 % recovery) were 16:0 fatty acid methyl ester (FAME) (32.12 %), 18:1cis 11 dimethyl acetal (DMA) (16.47 %), 16:1cis 9 DMA (10.28 %), 16:1cis 9 FAME (8.10 %), and 18:1cis 9 DMA (5.36 %). On the basis of morphological, physiological, and phylogenetic data, Clostridium geopurificans is proposed as a new species in genus Clostridium, with strain MJ1(T) as the type strain.

Consorcios microbianos con actividad ácido-láctica promisoria aislados desde inoculantes bacterianos nativos para ensilajes

&lt;p&gt;El objetivo de este trabajo fue aislar y caracterizar cepas de bacterias ácido lácticas (BAL) a partir de preparados microbianos (PM) nativos elaborados a base de residuos agroindustriales, como suero de leche, estiércol bovino y contenido ruminal, y diseñados como inoculantes biológicos para procesos de ensilaje de residuales orgánicos poscosecha. Se evaluaron 5 tratamientos (preparados microbianos, PM + 33% de residuo inoculante), con 3 réplicas: 1) Tratamiento control sin inoculante (PMSI), 2) Suero de leche (PMSL), 3) Estiércol bovino (PMEB), 4) Combinación de suero+estiércol (PMM) y 5) Contenido ruminal (PMCR); además, en todos se adicionó: melaza (20%), urea (1%), sal mineral (1%) y agua (45%). En recipientes plásticos se fermentaron tapados durante 96 horas, a 14 °C; de allí se obtuvieron 3 muestras de 500 mL de cada PM, se sembraron en agar MRS (Man, Rogosa y Sharpe) y se incubaron a 37 ºC durante 72 h bajo condiciones anaerobias (AnaeroGen, Oxoid). Las cepas de BAL son catalasa y oxidasa negativas, cocos, coco-bacilos y bacilos Gram positivos no esporulados. La identificación bioquímica se realizó con el sistema API-50CHL (BioMérieux, France) y la base de datos APILAB Plus versión 3.3.3. Se aislaron 39 cepas de BAL, 4 del PMSI (2 Lactobacillus plantarum, 1 L. brevis y 1 Lactococcus lactis), 12 del PMSL (3 Lactobacillus rhamnosus, 3 L. casei, 2 L. acidophilus, 2 L. paracasei y 2 L. fermentum), 7 del PMEB (3 Streptococcus bovis, 2 Lactobacillus plantarum y 2 Leuconostoc mesenteroides), 10 del PMM (3 Lactobacillus casei, 2 L. plantarum, 2 L. acidophilus, 1 L. fermentum, 1 L. rhamnosus y 1 L. paracasei) y 6 del PMCR (2 Lactobacillus vitulinus, 2 L. ruminus, 1 L. reuteri y 1 Pediococcus damnosus). En conclusión, la mejor fuente de cepas de BAL fue PMSL, que le otorga ventajas como posible inoculante bacteriano para ensilajes. En todos los PM se identificaron cepas homofermentativas y heterofermentativas, asociadas en consorcios microbianos; se generó una alternativa tecnológica para aprovechar tres residuos agroindustriales.&lt;/p&gt;