Open Access Original Research Article

Unusual Oropharyngeal Asymptomatic Manifestations Caused by Atypical Pathogens Detected by PCR into Altered Ecosystems of an Infertile Couple

Giovanni Di Bonaventura, Marco Del Boccio, Alfonso Pennelli, Marcello Rapinese, Raffaele Tenaglia, Pier Enrico Gallenga, Giampiero Neri, Gilberto Del Boccio

Microbiology Research Journal International, Page 447-458
DOI: 10.9734/BMRJ/2015/13082

The young couple casually arrived at our observation presented two different and painless altered ecosystems. The female partner exhibited inflamed lingual surface, while the man manifested several genital ulceration areas. Microbiological analysis of several sites belonging to each ecosystem revealed a latent dismicrobic ecosystem, as also confirmed by altered pH values. Interestingly, microbiological analysis for Mycoplasma hominis, Ureaplasma urealyticum, and Chlamydia trachomatis gave different results in both ecosystems, highlighting that each partner was differently infected by the same “silent infections”. The frequency of Ear Nose and Throat (ENT) chronic morbidity for these atypical infections increases, in the couple, the probability that mycoplasmosis and chlamydiasis serve as a reservoir of infection and/or reinfection for the urogenital tract. Although it is well known that several common bacteria and “atypical pathogens” - like mycoplasmas and C. trachomatis - can colonize both oral and urogenital ecosystems, rarely happens that they can clinically manifest their presence by unusual clinical manifestations we observed in different ecosystems investigated, hiding the unique true problem of infertility in the young couple. In this regard, our results showed that only starting from an accurate anamnesis, using an adequate sampling modality, adopting a new procedure to quantify microbial populations into asymptomatic ecosystems, and introducing the constant use of PCR analysis, it will be possible to disclose similar precocious problems of couple infertility, in genetically predisposed subjects.

Open Access Original Research Article

Influenza and Respiratory Syncytial Virus in Hospitalized Patients with Acute Exacerbation of Severe COPD in Izmir, Turkey

Gulrü Erbay, Ceyda Anar, Gunes Senol, Can Bicmen, Yusuf Pinar, Melih Buyuksirin

Microbiology Research Journal International, Page 459-465
DOI: 10.9734/BMRJ/2015/13251

Aims: The aim of this study is documenting the presence of respiratory syncytial virus (RSV) and influenza virus infections in hospitalized patients with AE-COPD in autumn and winter months.
Study Design: The study was designed as a prospective cohort study.
Place and Duration of Study: September 2006 and February 2007, Izmir, Turkey.
Methodology: Severe AE-COPD patients were studied by ready to use real time-PCR system and micro ELISA RSV and Influenza IGM and IgG antibody detection systems.
Results: Fifty AE-COPD cases enrolled to the study. Influenza and RSV were detected in 13 (26%) and 12 (24%) patients, respectively. Seven (14%) bacterial agents (4 P. aeruginosa, 2 S. pneumoniae, E. coli) were isolated from patients with viral infections.
Conclusion: Influenza viruses and RSV are important causative viral agents of AE-COPD. PCR and serological methods were decided to be used together for diagnosis of viral AE-COPD.

Open Access Original Research Article

Influence of UV Mutagenesis on β-Mannanase Production Potential of Aspergillus glaucaus and Rhizopus japonicus

D. J. Arotupin, B. J. Akinyele, O. O. Olaniyi

Microbiology Research Journal International, Page 466-473
DOI: 10.9734/BMRJ/2015/6762

Aim: The present study was conducted to improve the β-mannanase production from Aspergillus glaucausand Rhizopus japonicus using UV mutation.
Study Design: The first experiment, spore suspensions of A. glaucaus and R. japonicus was exposed to UV irradiation, while in the second experiment, wild types and mutant strains of A. glaucaus and R. japonicuswere screened for β-mannanase production in submerged fermentation.
Place and Duration of Study: Microbiology Research Laboratory Federal University of Technology, Akure, Ondo State, Nigeria between July and August, 2012.
Methodology: Mutants of A. glaucaus and R. japonicus were generated by exposure of spores suspension to UV irradiation for a period of 110 minutes at a distance of 13 cm in dark from the centre of germicidal lamp (240 nm) at 10 min intervals, 1 ml spores suspension was withdrawn and plated on Malt Extract Agar (MEA). The developed mutants and wild parents were screened for β-mannanase production in submerged fermentation. Quantitatively, β-mannanase activity was determined using dinitrosalicylic acid method, while protein content was determined by Lowry method.
Results: Eleven UV mutant fungal strains were generated for each of the wild types (A. glaucaus and R. japonicus) within 110 min of spore exposure to UV irradiation. The amount of enzyme produced by the mutants varied with the time of exposure. Approximately 27% of the mutant of A. glaucaus (9A1UV30, 9A1UV50 and 9A1UV70) generated from 30, 50 and 70 min of exposure to UV irradiation showed higher increase in β-mannanase activities when compared with parent strain, while repression of enzyme biosynthesis was observed in other mutants. Of all the mutants generated, the 9A1UV30 mutant had the highest increase in mannanase activity with approximately 46% higher than the parent strain, while the mutant 9A1UV10 exhibited 0% enzyme activity. Beta-mannanase production potential was repressed in the mutants of R. japonicus except for mutant 9A2UV50 where unappreciable higher increase of enzyme activity of 100.90% was observed in comparison with the parent strain.
Conclusion: Enhanced β-mannanase production was obtained from mutant strains 9A1UV30, 9A1UV50 and 9A1UV70 of A. glaucaus and they could be exploited commercially for industrial production of β-mannanase to meet industrial demand. To the best of my knowledge, this is the first report on successful mannanase producer mutants of A. glaucaus and R. japonicus and it is suggested that molecular studies should be carried out on the improved mutants to reveal the mutation.

Open Access Original Research Article

Detection of Salmonella spp. and Microbiological Analysis of Milk and Milk Based Products Available within Dhaka Metropolis, Bangladesh

Sharmina Yasmin, Shahana Parveen, Md. Sakil Munna, Rashed Noor

Microbiology Research Journal International, Page 474-480
DOI: 10.9734/BMRJ/2015/11010

Aims: With a previous observation of pathogenic drug-resistant microorganisms in raw milk samples, current investigation further endeavored to determine the presence of Salmonella spp. and portrayed a complete map of the overall microbiological feature of milk and milk based products available within Dhaka metropolis, Bangladesh.
Methodology: Conventional cultural and biochemical methods were carried out to isolate and enumerate the milk and milk based products accessing microorganisms. Standard agar disc diffusion method was conducted to assess the anti-bacterial resistance pattern of the isolated Salmonella spp.
Results: Out of 35 samples studied, 16 were found to be contaminated with E. coli (~106 cfu/ml) and 9 samples were found to be contaminated with Salmonella spp. (~105 cfu/ml). Fungal growth was also observed in all samples (~106). The Salmonella isolates were found to be resistant against ampicillin (10 µg), cotrimoxazole (25 µg), chloramphenicol (30 µg), ciprofloxacin (5 µg), cefixime (5 µg) and sensitive against ofloxacin (5 µg).
Conclusion: As projected from the current study, the proliferation of Salmonella spp. along with their drug-resistance traits could impart the morbidity and mortality associated risk to the overall public health safety.

Open Access Original Research Article

Influence of Cultural and Nutritional Factors on β-Mannanase Production by Penicillium italicum under Submerged State Fermentation

O. O. Olaniyi, T. C. Ekundayo, O. F. Igbe, Y. D. Fagbohungbe

Microbiology Research Journal International, Page 481-489
DOI: 10.9734/BMRJ/2015/6887

Aims: The process parameters influencing enzyme production were optimized to ascertain the optimal cultural and nutritional conditions for β-mannanase production by Penicillium italicum in submerged state fermentation.
Study Design: Five stages of experimental processes were designed for this study. The first stage, samples were withdrawn after 24, 48, 72, 96, 120, 144,168 and 192 h incubation. The second experiment, different agricultural wastes were screened as substitute to Locust Bean Gum. In third experiment, the effect of different pH values on β-mannanase production was evaluated, while the fermentation media were incubated at different temperatures in the fourth experiment. In fifth experiment, the effect of different surfactants on β-mannanase activity was evaluated.
Place and Duration of Study: Microbiology Research Laboratory, Federal University of Technology, Akure Nigeria between September 2011 and March 2012.
Methodology: Beta-Mannanase production was conducted in mineral salt medium and enzyme activity determined by dinitrosalicylic acid method, while protein content was determined by Lowry method.
Results: Maximum β-mannanase activity (41.667 U/ml) was observed after 120 h of incubation. Different agricultural wastes were screened as carbon substrates for β-mannanase production. Among tested carbon sources, orange peels proved to the best for β-mannanase production with an activity of 50.000 U/ml. Initial pH of the culture medium was optimized and a pH of 6.0 (130.556 U/ml) was found to be the best pH for β-mannanase activity. The optimum temperature was 30°C with an activity of 136.418 U/ml. Of all surfactants screened, Tween 20 (67.500 U/ml) gave the highest β-mannanase activity.
Conclusion: The nutritional and cultural factors obtained to be optimal from this study will help to design an experiment for maximum mannanase biosynthesis using cheaper substrates in place of commercial substrates known to be expensive for enzyme production.

Open Access Original Research Article

Frequency of Extended-Spectrum Beta-Lactamase (ESBLs) –Multidrug Resistance Produced by Pseudomonas aeruginosa Isolated from Clinical Specimens in Khorramabad City, Iran

Hossein Azizi, Farnaz Kheirandish, Mohsen Mirzaei

Microbiology Research Journal International, Page 490-495
DOI: 10.9734/BMRJ/2015/13360

Background: Pseudomonas aeruginosa is one of the most important pathogenic bacteria causing hospital infections, which has intrinsic resistance to many antibiotics. One of the reasons for the emergence of drug resistance in P. aeruginosa isolates is the production of ESBLs (extended-spectrum beta-lactamase) enzymes. Resistance rate is increasing due to the production of these enzymes in P. aeruginosa. The goal of this study was to determine the prevalence of ESBL senzymes in P. aeruginosa isolates from different samples of patients in Khorramabad city by CDT (combined disk test) phenotypic method.
Methods: This study was an investigation on 70 P. aeruginosa samples isolated from patients in medical centers of Khorramabad city during one year (2013). The bacteria were identified by routine biochemical tests. Antibiotic sensitivity of the isolates was evaluated by double disk diffusion method. Phenotypic investigation of ESBLs production among the tested isolates using cefotaxime and ceftazidime disks alone and in combination with clavulanic acid (combined disk test) was performed.
Results: Results of combined disk test showed that, out of 70 isolated pseudomonas, 100% of the samples had multi-drug resistance. Maximum resistance to cefixime (79.1%) and minimum resistance to meropenem (25%) were observed. Out of 70 isolates, 25 (35.7%) were phenotypic beta-lactamase-producing enzymes. That highest percentage was related to wounds, equal to 12 (17%).
Conclusion: Considering the increasing prevalence of P. aeruginosa strains which produce extended-spectrum beta-lactamase, it is recommended to use an appropriate treatment protocol based on determining antibiogram pattern of strains. This means that all isolated before treating with antibiotics were examined by antibiotic susceptibility test. Also antibiotics should be used according to CLSI (Clinical and Laboratory Standards Institute). This issue is of serious concern for applying infection-control criteria with the purpose of preventing spread of this microbe.