Cultivable bacterial diversity from the human colon

Knowledge of the composition of the colonic microbiota is important for our understanding of how the balance of these microbes is influenced by diet and the environment, and which bacterial groups are important in maintaining gut health or promoting disease. Molecular methodologies have advanced our understanding of the composition and diversity of the colonic microbiota. Importantly, however, it is the continued isolation of bacterial representatives of key groups that offers the best opportunity to conduct detailed metabolic and functional studies. This also permits bacterial genome sequencing which will accelerate the linkage to functionality. Obtaining new human colonic bacterial isolates can be challenging, because most of these are strict anaerobes and many have rather exact nutritional and physical requirements. Despite this many new species are being isolated and described that occupy distinct niches in the colonic microbial community. This review focuses on these under-studied yet important gut anaerobes.     

Investigation of the internal bacterial flora of Eurygaster integriceps (Hemiptera: Scutelleridae) and pathogenicity of the flora members

Eurygaster integriceps (Hemiptera: Scutelleridae) is one of the most serious pest of wheat and other cereal crops throughout the Near and Middle East including Turkey. To contribute biological control of this pest, we investigated the culturable bacterial flora of E. integriceps and their virulence against it. We also tested different entomopathogenic fungi (Isaria fumosorosea ARSEF8356, Beauveria bassiana ARSEF8356, Metarhizium brunneum ARSEF8671 and Nomurae rileyi ARSEF1670) against this pest under laboratory conditions. Bacterial isolates were characterized based on morphological, biochemical, physiological and molecular characteristics (16S rRNA sequencing). The isolates were identified as Pantoea sp. (S1, S5, S7, S8, S10 and S11), P. agglomerans (S2, S3 and S4), Pseudomonas sp. S6 and Micrococcus luteus S9. The highest mortality within bacterial isolates was observed from Pantoea sp. S1, P. agglomerans S4 and Pantoea sp. S7 with 100%. Mortality of other bacterial species ranged from 33% to 88%. The highest mortality among the tested entomopathogenic fungi was obtained from Isaria fumosorosea ARSEF 8333 with 100%. Mortality of other fungi ranged from 33% to 50%. Consequently, Pantoea sp. S1, P. agglomerans S4, Pantoea sp. S7, Beauveria bassiana ARSEF 8356 and Isaria fumosorosea ARSEF 8356 seem to be promising candidates in the control of E. integriceps.     

A comparison of bacterial flora isolated by transtracheal aspiration and pharyngeal swabs in Macaca fascicularis.

The pulmonary flora of 30 monkeys (Macaca fascicularis) was sampled by the transtracheal aspiration technique and the pharyngeal swab method, and the results were compared. The transtracheal aspiration technique yielded lower numbers of bacteria in both aerobic and anaerobic cultures. The bacteria isolated by transtracheal aspiration were predominately pure culture, thereby lowering the possibility of contamination from commensal flora. Bordetella bronchiseptica was isolated from 23.3% of the monkeys by transtracheal aspiration, but this organism was not isolated when samples were collected with pharyngeal swabs.     

Composition of the bacterial flora in tonsilloliths

Tonsilloliths are a potential cause of oral malodor. In this study, microbial profiles and composition of tonsilloliths were determined using culture-independent molecular methods and scanning electron microscopy. 16S ribosomal RNA bacterial genes (16S rDNAs) isolated from tonsilloliths of 6 individuals were amplified by PCR and cloned into Escherichia coli. Partial 16S rDNA sequences of approximately 600 bases of cloned inserts were used to determine species identity by comparison with sequences of known species. Characteristics of bacteria on the surface and inside the tonsillolith were analyzed using scanning electron microscopy. Anaerobic bacteria detected in tonsilloliths belonged to the genera Eubacterium, Fusobacterium, Megasphaera, Porphyromonas, Prevotella, Selenomonas and Tannerella, all of which appear to be associated with production of volatile sulfur compounds. Electron microscopy revealed cocci and rods on the surface and rods predominating inside the tonsilloliths. These results support the tonsillolith as an origin of oral malodor.     

Superintendence of antimicrobial resistance observed in bacterial flora isolated from human faecal carriage in Vellore,India

A frequent cross-sectional study was conducted to determine the patterns of antimicrobial resistance in 296 bacterial strains isolated from in-patient faecal samples of Government Vellore Medical College and Hospital, Vellore. Isolation and identification of bacterial strains were done using enrichment media, selective media, and biochemical tests. Antimicrobial susceptibility testing by the disc diffusion method and minimal inhibitory concentration method was conducted and the strains were subjected to extended spectrum beta-lactamases screening. Antibiotic sensitivity pattern of Staphylococcus spp. showed oxacillin resistance. Almost all the strains were sensitive to linezolid, vancomycin, gentamycin and chloramphenicol. In gram negative isolates ciprofloxacin and tobramycin showed better sensitivity and ceftazidime showed a higher percentage of resistance by MIC. Out of 250 isolates, Enterobacteriaceae showed positive for 86/250, 82/250 and 94/250 isolates and 3/10, 4/10 and 4/10 non-Enterobacteriaceae isolates were found to be positive for CTX-M gene, TEM gene and SHV gene, respectively. This study helps to assess/analyse the relation between the spectrum of microorganisms present in various grades of faecal carriage and their susceptibility pattern in this part of the Vellore town.     

Monitoring the diel activity of Ixodes ricinus ticks in Hungary over three seasons

Ixodes ricinus is the most common tick species in Europe and vector of many diseases of humans. The risk of contracting such a disease is influenced by many factors, but one of the crucial points is questing activity of unfed ticks. In order to supplement the few literature data on patterns of diel activity of this tick species and to examine the correlations between data on diel activity of ticks and their small mammal hosts and some meteorological variables, a survey was performed. Diel activity of questing I. ricinus and small rodents was studied in a known natural tick-borne encephalitis virus focus over 7 months at one sampling day monthly. 1,063 I. ricinus ticks and 25 rodents were sampled. Air temperature and humidity data were also recorded in the 24 study plots at time of sampling. From April to October questing activity of nymphs increased in the 3-h-period after sunrise, comparing to activity of the 3 h before sunrise. Proportion of nymphs sampled 3 h after sunset compared to total sampled nymphs 3 h before and 3 h after sunset showed correlation to activity of rodents. In the period of April–July both nymphs and larvae showed stronger activity from sunrise to sunset, this turned to dominant nighttime activity in August–September, whereas activity changed to equal in day and night in October. Our results indicate that natural light and rodent population have positive effect on questing activity of I. ricinus.     

Maternal diseases and isolated orofacial clefts in Hungary

BACKGROUND: Isolated orofacial clefts (OFCs) are likely to be caused by gene-environment interaction; therefore, the objective of the current study was to evaluate the possible association between all maternal diseases during pregnancy and isolated cleft lip with or without cleft palate (CL+/-CP) and posterior cleft palate (PCP) in the offspring. METHODS: The database of the large population-based Hungarian Case-Control Surveillance of Congenital Abnormalities, 1980-1996, was evaluated. The database includes 1374 cases with isolated CL+/- CP and 601 with PCP, plus 38,151 matched population controls (without defects) and 20,868 patient controls with other defects. Data collection was based on prospective medical records, retrospective maternal data via a self-reported questionnaire, and home visits of nonresponding families. RESULTS: An increased risk for isolated CL+/- CP was found for children born to mothers with influenza, common cold, orofacial herpes, and gastroenteritis during pregnancy. Risk for isolated PCP was increased in children of mothers with influenza, sinusitis, and bronchitis. Among chronic maternal diseases, epilepsy and angina pectoris showed a higher prevalence in the mothers of children born with isolated OFCs (cases). CONCLUSIONS: Some maternal diseases are risk factors for the pathogenesis of isolated OFCs. It is worth considering the prevention of possible harmful effects of influenza by vaccination during the expected epidemic period.     

Mycological flora isolated from people in Poland

It has been shown that in Poland the most frequently encountered pathogenic fungi are the zoophilic ones:T. mentagrophytes, M. canis andT. verrucosum. The zoophilic fungi have been isolated in 69.6 % of cases, while the anthropophilic fungi in 30.3 % of cases. In Poland, the most frequently isolated fungus among anthropophilic fungi have beenT. rubrum (36.2 %) andT. violaceum (34.3 %), whereasT. schoenleinii has been found in 5.7 % of cases. The geographical distribution of the anthropophilic fungi indicates that they appear most frequently in the counties of Biaystok (58.2 %), Katowice (40.5 %), and Warsaw (36.5 %), while the zoophilic fungi are found prevalently in the counties of ód (93.0 %), Kielce (84.7 %) and in that of Szczecin (83.8 %). It has been ascertained that the diseased animals and sick persons form the main reservoirs of the pathogenetic fungi. For the prophylactic purposes it is indispensable to promote among the population the sanitary instructions and propagate the principles of personal hygiene, which play a very important role in preventing mycosis. Within the framework of antimycosis campaigns it is necessary to widen the collaboration between the dermatologists and veterinary surgeons.Summary The author came to the conclusion that people in Poland suffer mostly of zoo-philic (69.9 %) than of anthropophilic dermatomycoses (30.3 %).Trichophyton rubrum was observed in 36.2 %,T. violaceum in 24.3 % andT. schoenleinii in 5.7. %. The main reservoir of dermatomycoses in Poland are sick animals and people. In the fight against mycoses of people and animals the collaboration of dermatologists and veterinarians is absolutely necessary.

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Résumé L'auteur a démontré qu'en Pologne chez les hommes la fréquence des dermatomycoses zoophiles est plus haute (69.7 %) que celle des dermatomycoses anthropophiles (30.3 %). Trichophyton rubrum noté dans 36.2 %,Trichophyton violaceum dans 24.3 % etTrichophyton schoenleinii dans 5.7 % cas. Les réservoirs principaux des mycoses en Pologne sont les animaux et les hommes malades. Dans la lutte contre les mycoses chez les hommes et chez les animaux, il est indispensable de constituer une collaboration stricte entre les dermatologues et les véterinaires.

     

细菌性阴道病肠道菌群与阴道菌群16S rDNA测序分析

目的通过对细菌性阴道病(BV)患者肠道菌群及阴道菌群16S rDNA扩增子测序,分析其结构、多样性、相关性以及BV对肠道菌群的影响,为今后治疗BV提供新的思路。方法选取符合纳入标准的BV患者11例(BV组),健康者9例(C组),留存阴道分泌物及新鲜粪便进行16S rDNA基因检测分析。结果C组阴道菌群以乳杆菌属为主,BV与加德纳菌属、普雷沃菌属、Sneathia、窄食单胞菌属(Stenotrophomonas)、阿托波菌属、Shuttleworthia、巨型球菌属密切相关。BV组肠道、阴道菌群丰富度均高于C组。Alpha多样性分析中C组和BV组肠道菌群、阴道菌群的Shannon指数组间比较,χ²值为29.137, P=0.000<0.05,两组阴道菌群Shannon指数组间比较差异具有统计学意义(P<0.05),BV组高于C组。物种多样性曲线反映本研究样本测序数据量的合理性,表明BV组的肠道菌群多样性、丰富度均高于C组,主坐标分析表明C组肠道与阴道的菌群结构差距较大,BV组肠道与阴道的菌群结构有相似之处,且两组肠道菌群结构接近。BV组阴道菌群中厚壁菌门丰度较C组低,放线菌门、拟杆菌门较C组高;BV组肠道菌群中拟杆菌门丰度较C组低;C组肠道中拟杆菌门明显高于阴道,厚壁菌门明显低于阴道; BV组阴道菌群中放线菌门丰度高于C组,差异均具有统计学意义(P<0.05)。结论BV阴道菌群与肠道菌群具有相关性,BV可能引起肠道菌群结构比例和多样性的改变。     

Influence of biotope on the distribution and peak activity of questing ixodid ticks in Hungary

Abstract In order to update the occurrence of hard tick species in Hungary, 3442 questing ticks were collected from the vegetation by the dragging/flagging method in 37 different places in the country, between March and June of 2007. Ixodes ricinus (L.) turned out to be ubiquitous. Dermacentor marginatus (Schulzer) was absent from sampling sites in the southwestern part of the country, but in most places was concomitant and contemporaneous with Dermacentor reticulatus (Fabricius). These two species, as well as I. ricinus, occurred up to an altitude of 900–1000 m a.s.l. Haemaphysalis inermis (Birula) and Haemaphysalis concinna (Koch) were not confined to any parts of the country, unlike Haemaphysalis punctata (Canestrini & Fanzago) which was found in only one region. The local prevalence of the latter species was also significantly lower than those of the former two in the same habitat (fringes of meadows, paths in forests). Dermacentor spp. and H. inermis were represented only by adults. In most species females were collected more frequently than males, except in H. concinna and H. punctata. Temporal differences between the peak activity of I. ricinus and Dermacentor spp. on dry pastures appeared to equalize on meadows in mountain forests, and a similar phenomenon was observed for the three Haemaphysalis spp. when collected along forest paths with fresh, green vegetation.     

Coexistence of emerging bacterial pathogens in Ixodes ricinus ticks in Serbia

The list of tick-borne pathogens is long, varied and includes viruses, bacteria, protozoa and nematodes. As all of these agents can exist in ticks, their co-infections have been previously reported. We studied co-infections of emerging bacterial pathogens (Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Francisella tularensis) in Ixodes ricinus ticks in Serbia. Using PCR technique, we detected species-specific sequences, rrf-rrl rDNA intergenic spacer for B. burgdorferi s.l., p44/msp2 paralogs for A. phagocytophilum, and the 17 kDa lipoprotein gene, TUL4, for F. tularensis, respectively, in total DNA extracted from the ticks. Common infections with more than one pathogen were detected in 42 (28.8%) of 146 infected I. ricinus ticks. Co-infections with two pathogens were present in 39 (26.7%) of infected ticks. Simultaneous presence of A. phagocytophilum and different genospecies of B. burgdorferi s.l. complex was recorded in 16 ticks, co-infection with different B. burgdorferi s. l. genospecies was found in 15 ticks and eight ticks harbored mixed infections with F. tularensis and B. burgdorferi s.l. genospecies. Less common were triple pathogen species infections, detected in three ticks, one infected with A. phagocytophilum / B. burgdorferi s.s. / B. lusitaniae and two infected with F. tularensis / B. burgdorferi s.s. / B. lusitaniae. No mixed infections of A. phagocytophilum and F. tularensis were detected.     

Some components of preputial bacterial flora in the ram

During the reproductive period the AA have carried on bacteriological researches in the preputial cavity in eight sardinian rams, without pathological processes, living in eight flocks in the country around Sassari, in order to detect the bacteriological flora. This flora could interfere both with fertility and with conservation of seminal material for artificial insemination. Three withdrawals have been carried on before, during and after the reproduction season. E.coli, P. mirabilis, Staph. epidermidis have been found in the varius examined subjects. The AA stress that the same bacterium which had been isolated in the first withdrawal. Moreover in some subjects the bacteriological flora had increased. The AA stress how these data can be useful in order to control the flock fertility and in order to improve the technology of conservation of seminal material.