Advanced Journal of Microbiology Research

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Penicillium verruculosum RS7PF: A root fungal endophyte associated with an ethno-medicinal plant of the indigenous tribes of Eastern India

R. K. Bhagobaty, S. R. Joshi* and R. Kumar

Microbiology Laboratory, Department of Biotechnology and Bioinformatics, North Eastern Hill University, Umshing, Shillong-793 022, Meghalaya, India.

Accepted 11 March, 2015

Abstract

Potentilla fulgens L., an ethno-medicinal plant of the ethnic tribes of Meghalaya in the Eastern Himalayan range of India was screened for root fungal endophytes. This plant is distributed in the wild habitats and in the traditionally preserved ‘Sacred Groves’ of Meghalaya in Eastern India, which is of late getting over-exploited for its medicinal value. Isolation of endophytes in water agar plates yielded a unique fungus that had similar morphological characteristics with the members of the genus Talaromyces. Scanning and transmission electron microscopic studies of the fungus revealed its surface morphological features, close association and extensive colonization of plant root tissues. Molecular characterization using fungal specific 18S rRNA primers showed that the fungus shared a 98% homology with Penicillium verruculosum (Genbank accession number AF510496). The isolate investigated is likely to be a novel endophytic strain of P. verruculosum which has evolved a very close symbiotic association with the host plant roots. The fungal endophyte characterized is being explored as an early colonizer in the establishment of seedlings and successful micropropagation of the plant.

Key words: Potentilla fulgens L., ethnic tribes, Sacred Grove, fungal endophyte, molecular characterization, Penicillium verruculosum.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Comparison of extract bio-activities of in-situ and in vitro grown selected bryophyte species

Aneta Sabovljevi ¹, Marina Sokovi ², Jasmina Glamo lija³, Ana  iri ², Milorad Vuji i ¹, Boris Pejin³ and Marko Sabovljevi ¹*

¹Faculty of Biology, Institute of Botany and Botanical Garden, University of Belgrade, Takovska 43, 11000 Belgrade, Serbia.

²Institute for Biological Research “S. Stankovic”, Bul. Despota Stefana 142, 11000 Belgrade, Serbia.

³Faculty of Chemistry, University of Belgrade, Studentski trg 16, 11000 Belgrade, Serbia.

Accepted 26 February, 2015

Abstract

The antimicrobial activity of DMSO extracts of three bryophyte species, two mosses and a liverwort (Atrichum undulatum (Hedw.) P. Beauv., Marchantia polymorpha L. ssp. ruderalis Bischl. and Boisselier, Physcomitrella patens (Hedw.) Bruch and Schimp.) grown in nature and in axenic culture was evaluated by microdilution method against eight bacterial species ( Escherichia coli ATCC 35210, Pseudomonas aeruginosa ATCC 27853, Salmonella typhimurium ATCC 13311, Enterobacter cloacae (human isolate), Listeria monocytogenes NCTC 7973, Bacillus cereus (human isolate), Micrococcus flavus ATCC 10240 and Staphilococcus aureus ATCC 6538). All investigated bryophyte extracts are proved to be active against all bacteria tested. In general, extracts made from material grown in nature express better anti- bacterial activity comparing to those made from material grown in laboratory conditions. Some of the bacteria react the same to both extracts and some even better to the extracts made from axenically grown material.

Key words: Bryophytes, antibacterial activity, Atrichum undulatum, Marchantia polymorpha, Physcomitrella patens.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

The Copepod parasite of the gills of four teleost fishes caught from the gulf of Annaba (Algeria)

Chahinez Boualleg¹, Hadda Ferhati¹, Nouha Kaouachi¹, Mourad Bensouilah¹ and Sonia Ternengo²

¹Laboratoire d’Ecobiologie des Milieux Marins et Littoraux, Faculté des Sciences, Université Badji Mokhtar Annaba, BP: 12 El- Hadjar, Annaba 23000, Algérie.

²Laboratoire (Parasites et écosystèmes méditerranéens), Faculté des Sciences et Techniques, Université de Corse, CNRS UMR 6134, BP 52, 20250, Corte, France.

Accepted 15 April, 2015

Abstract

The examination of the gills of 960 fishes belonging to four species: Diplodus annularis, Pagellus erythrinus, Lithognatus mormyrus and Mullus barbatus collected from the Gulf of Annaba, enabled us to identify 13 species of parasitic copepods: Caligus ligusticus, Caligus diaphanus, Clavellotis sp, Clavellotis sargi, Clavellotis pagri, Clavellotis strumosa, Lernaeolophus sultanus, Hatschekia pagellibogneravei, Hatschekia sp, Hatschekia mulli, Sparidicola lithognathi, Neobrachiella exigua and Alella macrotrachelus. The distribution of copepods identified, varies from one host species to another and one season to another. P. erythrinus and L. mormyrus present the important number of copepods species. However, the winter presents the highest parasitic diversity (13). Furthermore, the majority of oixenous species are counted. The rates of most values are recorded in summer (33%) and spring (25%). It is also, D. annularis which shelters more of the 2/3 population of copepods collected. The copepod H. pagellibogneravei is the most abundant. The evaluation of parasitic indices shows that, it is during the summer and spring that the maximum values are recorded.

Key words: Copepod gills, Gulf of Annaba, teleost fish, season.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Application of PCR-RFLP of gap gene method as a molecular typing tool for coagulase negative Staphylococci from bovine and human origin identified with VITEK 2

Emel Banu Buyukunal Bal¹*_, Mehmet Ali Bal², Taner Isevi² and Erkan Yula³

¹Department of Biology, Faculty of Science and Letters, Kahramanmaras Sutcu Imam University, Kahramanmaras, Turkey.

²Department of Animal Science, Faculty of Agriculture, Kahramanmaras Sutcu Imam University, Kahramanmaras, Turkey.

³Department of Microbiology and Clinical Microbiology, Faculty of Medicine, Cukurova University, Adana, Turkey.

Accepted 2 April, 2015

Abstract

The aim of this study was to apply the Restriction Fragment Length Polymorphism (RFLP) of Glyceraldehyde-3-Phosphate Dehydrogenase encoding gene (gap) for testing its performance as a molecular typing tool in coagulase negative staphylococci (CNS) isolates from bovine mastitis (n = 59) and human clinical cases (n = 13) identified with VITEK 2. According to the phenotypic identification results, bovine mastitis isolates were Staphylococcus haemolyticus, Staphylococcus simulans, Staphylococcus auricularis, Staphylococcus warneri, Staphylococcus hominis, Staphylococcus capitis, Staphylococcus xylosus, Staphylococcus epidermidis and Staphylococcus cohnii. Although most of those isolates were generated PCR amplicons with gap gene specific primers, PCR amplification of gap gene failed in 29 from 72 isolates. The samples that did not produce amplicons were reamplified with Staphylococcal 16S rRNA gene specific primers. After PCR amplifications, amplicons were produced in 17 from 29 samples. Three different restriction endonucleases (AluI, MseI and RsaI) were used for PCR-RFLP analysis, among these AluI has been found the most discriminatory power for identification in species. The results of the RFLP of gap gene provide a support for the misidentification problem associated with VITEK 2 system for S. simulans, S. auricularis and S. capitis species. Moreover, more frequent failure in gap gene amplification for bovine isolates which were phenotypically identified as S. simulans, S. auricularis, S. capitis, S. xylosus and S. cohnii was not clear. In addition, the method verified the phenotypic identification for S. haemolyticus, S. warneri, S. hominis and S. epidermidis isolates with different rates at 100, 33.3, 57.1, and 66.7%, respectively.

Key words: Coagulase negative staphylococci, gap gene, PCR-RFLP.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Detection of adhesin genes and slime production among Staphylococci in orthopaedic surgical wounds

Nizami Duran¹*, Yunus Dogramaci², Burcin Ozer¹, Cemil Demir¹ and Aydiner Kalaci²

¹Department of Microbiology and Clinical Microbiology, Medical Faculty, Mustafa Kemal University, Hatay, Turkey.

²Department of Orthopaedics and Traumatology, Medical Faculty, Mustafa Kemal University, Hatay, Turkey.

Accepted 12 March, 2016

Abstract

This study was aimed at investigating: (i) three adhesin genes (clf A, fnb A and cna) in Staphylococus aureus strains, (ii) the presence of slime (ica A and ica D genes) in both Staphylococus epidermidis and S. aureus strains isolated from surgical wounds. The slime and adhesin genes were detected by multiplex PCR. The ica A/ica D positivity rates were determined as 66.2% (104/157) in a total of 157 staphylococcal strains. While the occurance rate of slime genes was 69.6% (48/69) among the S. epidermidis, this ratio was 63.6% (56/88) among the S.aureus isolates. No statistically significant difference was found between S. epidermidis and S. aureus isolates in terms of the presence of slime genes (p > 0.05). Among the 88 S. aureus strains, almost all of the strains were positive for fnb A gene (97.7%). The cna and clf A positivity rates were detected in 69 (78.4%) and 45 (51.1%) isolates, respectively. The ica A and ica D genes responsible for slime production have been found to have high prevalence. Also, the frequency of adhesin genes was determined at a high rate in S. aureus strains isolated from surgical wounds. Molecular identification of virulent staphylococcal strains may help in management in clinical decision making.

Key words: Adhesins, slime, Staphylococus aureus, Staphylococus epidermidis, wound infection.

Advanced Journal of Microbiology Research Vol. 2016

Available online at http://internationalscholarsjournals.org/journal/ajmr

© 2016 International Scholars Journals

Full Length Research Paper

Systematic position of Moesziomyces penicillariae among Ustilaginaceae

Gnagna Diagne-Leye¹*, Teresa Almaraz-Lopez^{2, 3}, Maïmouna Sy-Ndir¹, Gérard Mascarell⁴, Amadou Tidiane Ba⁵ and Christophe Roux^{2, 3}

¹Laboratoire de Biotechnologies des champignons (LBC), Département de Biologie Végétale, Faculté des Sciences et Techniques, Université Cheikh Anta Diop, B. P. 5005, Dakar, Senegal.

²Université de Toulouse, UPS, UMR 5546, Surfaces Cellulaires et Signalisation chez les Végétaux, B. P. 42617, F-31326, Castanet-Tolosan, France.

³CNRS, UMR 5546, B. P. 42617, F-31326, Castanet-Tolosan, France.

⁴Département Régulations, Développement et Diversité Moléculaire, Equipe « Cyanobactérie, Cyanotoxines, et environnement », MCAM, FRE 3206 CNRS/MNHN Paris, France.

⁵Département de Biologie Végétale, Faculté des Sciences et Techniques, Université Cheikh Anta Diop, B. P. 5005, Dakar, Senegal.

Accepted 15 April, 2015

Abstract

We revisited the systematic position of the fungus Moesziomyces penicillariae, the causal agent of smut of pearl millet, by using morphological characters, germination pattern of teliospores and molecular analysis of ribosomal sequences. Samples of smutted ears of pearl millet were harvested in Senegal (West Africa). Compared to the description of Moesziomyces genus sensus Vánky, our samples presented morphological differences: i) presence of a columella-like structure in sori; ii) surface ornamentations of teliospores; iii) teliospore germination similar to Ustilago and Sporisorium ones. We investigated the systematic position of our samples by aligning their Internal Transcribed Spacer (ITS) sequences of the ribosomal regions with 47 sequences from Ustilaginaceae. The resulting tree rooted with Tolyposporium junci allowed the separation of five groups among which, they are, two Ustilago and two Sporisorium. An independent clade is formed by Tranzscheliella williamsii and Tranzscheliella hypodytes species including Ustilago sparti. Moesziomyces species used in this analysis form a monophylletic group located in Ustilago 2 group, which include different Ustilago and Sporisorium species but also Pseudozyma antartica. Our results indicate the necessity to amend the Moesziomyces genus as the morphological and molecular data confirm that they are included in the Ustilago-Sporisorium complex.

Key words: Pearl millet smut, Moesziomyces penicillariae, Ustilaginaceae.