Open Access Short communication

First Insight into the Current Prevalence of Bovine Tuberculosis in Cattle Slaughtered in Cameroon: the Case of Main Abattoirs of Yaoundé and Douala

Francioli Koro Koro, Bouba ., Eric Foko, Alexandre François Ngatchou, Sara Eyangoh, François-Xavier Etoa

Microbiology Research Journal International, Page 272-279
DOI: 10.9734/BMRJ/2013/3065

Aim: The aim of this study was to evaluate the current prevalence of bovine tuberculosis at Yaoundé and Douala abattoirs.
Study Design: Many investigations confirmed that bovine tuberculosis is prevalent in cattle destined for consumption in Cameroon but the magnitude and the distribution of animal tuberculosis in the country are unknown.
Place and Duration of Study: Sampling was made during routine meat inspection, in the Yaoundé and Douala abattoirs located in the Central and Littoral regions of Cameroon respectively. Sampling was successively carried out from November 2010 to April 2011. Sample processing (culture, acid-fast staining and spoligotyping) was made at Mycobacteriology Reference Laboratory in Centre Pasteur of Cameroon.
Methods: About 16,316 slaughtered cattle, were successively inspected for tuberculosis during this study. Among them 9,127 and 7,189 were slaughtered in Yaoundé and Douala abattoirs respectively. Evidence of pathology was supported by postmortem examination of carcasses using visual examination and palpation of lungs, livers, hearts, internal bodies and lymph nodes. The prevalence was calculated as the number of cattle with suspected TB lesions divided by the number of cattle examined at post mortem within the specified period. Ziehl-Neelsen staining, culture on solid medium and spoligotyping were made to identify acid-fast Bacilli and Mycobacterium bovis.
Results: The overall apparent prevalence of bovine tuberculosis based on suggestive macroscopic lesions induced by tuberculosis was 1.03%. This prevalence was split to 0.81% and 1.3% in Yaoundé and Douala abattoirs respectively. Mycobacterium bovis accounted for 47.62% of the tuberculous lesions and its prevalence was 0.49%.
Conclusion: This result show that bovine tuberculosis is still prevalent in cattle destined for human consumption in Cameroon and highlighted the contribution of M. bovis as the leading cause of bovine tuberculosis in Cameroon.

Open Access Short communication

Infection of Aedes aegypti (Diptera: Culicidae) larvae and Adults by the Entomopathogenic Fungus Metarhizium anisopliae (Metschn.) Sorokin

Laerciana P. Vieira, Adriano R. Paula, Cátia O. Paula, Renato A. DaMatta, Richard I. Samuels

Microbiology Research Journal International, Page 309-317
DOI: 10.9734/BMRJ/2013/3638

Aims: The dengue vector Aedes aegypti is currently a target for control by entomopathogenic fungi. We have previously shown the susceptibility of both larvae and adult A. aegypti to fungal infection by Metarhizium anisopliae and strategies are being developed for control of both stages in the field.
Study Design: Few studies have been carried out on the early stages of the infection process of entomopathogenic fungi in mosquitoes, especially in the case of adult infection.
Place and Duration of Study: Department of Entomology and Plant Pathology and the Department of Cell Biology at the State University of North Fluminense. May 2010 – July 2011.
Methodology: Conidia were obtained for all experiments by culturing M. anisopliae in SDA. Conidial suspensions were standardized by serial dilution following quantification using a hemocytometer. Larvae were infected by addition of conidial suspensions to the water in which they were being maintained. Larvae were removed at specific time intervals for observation using a scanning electron microscope. Adult A. aegypti were infected by spraying with conidial suspensions at a concentration previously established. Following inoculation, mosquitoes were maintained in cages and cohorts were removed for observation using a scanning electron microscopy at specific time intervals.
Results: The initial stage of the infection process of larvae and adults (females) was described here. Conidia were found to attach to specific regions of the larvae, associated with thoracic hairs. Saprophytic fungal development was observed on the integument of larvae 36 hours after exposure to the fungus. For infection of adults, adhesion and germination of conidia were only observed on the thorax.
Conclusion: The pattern of fungal infection of mosquito larvae was different to that previously documented in the literature. The integument of the larvae was found to be a favorable environment for fungal development. This did not appear to be the case in colonization of the adult integument, which was a more hostile environment for the fungus. However, the microbial control of adult mosquitoes is thought to be more promising than that of larvae due to reduced half-life of the conidia in aqueous environments.

Open Access Minireview Article

Mini-Review: Theoretical and Onsite Evaluation of Hazard Potentials in the Local Production of Wara; an Indigenous West African Soft Cheese

S. I. B. Cadmus, O. M. Ajunwa, A. J. Adeleke, C. A. Agada, A. A. Awoyele

Microbiology Research Journal International, Page 218-234
DOI: 10.9734/BMRJ/2013/3147

The effects of food hazards have been felt in various forms over the years by consumers globally. This study attempts to assess and evaluate pertinent hazards inherent in the local production of wara (West African soft cheese) which is traditionally produced in small scale by Fulani women and consumed widely in Nigeria. The producers of wara in most cases are associated with poor hygiene, and their products are usually inconsistent in quality and possess very short shelf life. The processes of milking (collection of milk) from animal sources, transferring of the milk into metal pots for heating, heating of the milk, addition of coagulum, ladling of curds and whey into basket and transferring to container of cool water were identified as possible hazard entry points, which a properly designed quality control system should address. To check these hazards, the Hazard Analysis and Critical Control Points (HACCP) system has been proposed, and a major inclusion of this system in local production processes is advocated.

Open Access Original Research Article

Assessing the Microbiological Level and the Incidence of Water-soaking on the Proximate Composition of two Cultivars of Cowpea (Vigna unguiculata L.) Grains Grown in Côte d’Ivoire

Rose Koffi-Nevry, Marina Koussémon, W A Mireille Alloue-Boraud, Kouassi Kouassi

Microbiology Research Journal International, Page 206-217
DOI: 10.9734/BMRJ/2013/2511

Aims: This study was undertaken to assess the influence of seed treatment (soaking in water) on nutritional and microbiological composition of two cowpea cultivars.
Place and Duration of Study: Laboratory of Food Biochemistry and Tropical Products Technology, and the laboratory of Biotechnology and Food Microbiology, University of Nangui Abrogoua, Abidjan, between October 2010 and December 2011.
Study Design: Method based on AOAC tests and AFNOR for microbiological analysis. A two-way analysis of variance and t-test were used.
Methodology: The proximate composition of soaked and non soaked cowpea grains was determined and microbiological (bacteriological and mycological) analysis of these grains was also performed.
Results: The major components were 28% and 26.25% protein, 48.35% and 47.99% carbohydrate, 41.66% and 40.05% starch for the RC (red cultivar) and WC (white cultivar) respectively. Lipids are less represented in the 2 cultivars (2.5%). There were significant reductions in the contents of the major components as a result of the treatment. Plain water soaking brought about a significant decrease in the proximate composition causing a mean reduction of 3.14% and 10.02% protein, 28.23% and 29.30% carbohydrate, 29.47% and 28.94% starch, 18.80% and 22.02 % energy for the RC and WC respectively. The mean decrease for mineral was 23.13% and 47.66% iron, 2.32% and 8.15% calcium, 9.30% and 2.10% phosphorus for the RC and WC respectively. In general the highest reduction was observed in the WC variety. Mean count (Log10 cfu/g) of total aerobic miroflora, coliforms, mould and yeast were 6.29 and 6.43; 2.04 and 2.58; 4.41 and 4.78 for the RC and WC respectively. Five genera of mould were isolated: Aspergillus, Mucor, Penicillium, Botrytis and Geotrichum. The predominant fungi belonged to Aspergillus genus.
Conclusion: The cultivar types of cowpea and the preparation methods could affect the nutrient availability of this product. Cold water soaking has a great influence on the properties of cowpea grains.

Open Access Original Research Article

Can Combinations of Non-Invasive Parameters Replace Liver Biopsy in the Diagnosis of Hepatic Fibrosis in Egyptian Patients with Chronic Hepatitis C

Walid A. Abd EL Dayem, Mohammed Emam, Noha E. Shaheen, Mohamed H. Emara, Ehab M. Darweish, Nagla A. Abdelwahab

Microbiology Research Journal International, Page 259-271
DOI: 10.9734/BMRJ/2013/3323

Aims: Liver biopsy has always been represented as the standard reference for assessment of hepatic fibrosis although it has several limitations. This study aimed at evaluating the accuracy of noninvasive methods for diagnosis of hepatic fibrosis in adult Egyptian patients with chronic hepatitis C virus (HCV) infection.
Study Design: Cross sectional study.
Place and Duration of Study: This study was conducted in Al-Ahrar General Hospital (local treatment centre for Hepatitis C virus), Sharkia Governorate, Egypt and the Tropical Medicine Department, Zagazig University Hospitals, Sharkia Governorate, Egypt in the period from April 2011 to March 2012.
Methodology: Fifty chronic HCV patients were selected out of 255 chronic HCV patients awaiting assessment for combined pegylated interferon/ribavirin therapy according to the modified guidelines of the National Committee for Control and Prevention of viral Hepatitis C in Egypt. Diagnosis of HCV was confirmed by detection of anti-HCV antibody and positivity for HCV RNA for more than 6 months. All patients were assessed by liver biopsy and noninvasive methods namely aspartate transaminase/platelet ratio (APRI), abdominal ultrasonography measuring caudate/right lobe ratio and liver stiffness measurement.
Results: The accuracy in diagnosis of liver fibrosis using different methods in comparison to liver biopsy was 60%, 84%, 88%, 90%, 92% and 84% for APRI, ultrasonography, Fibroscan, combined Fibroscan/APRI, Fibroscan/ultrasonography and APRI/ultrasonography respectively. The sensitivity was 62.5%, 87.5%, 87.5%, 90.6%, 93.8 and 87.5 for APRI, ultrasonography, Fibroscan, combined Fibroscan/APRI, Fibroscan/ultrasonography and APRI/ultrasonography respectively. The specificity was 55.6%, 77.8%, 88.9%, 88.9%, 88.9 and 77.8 for APRI, ultrasonography, Fibroscan, combined Fibroscan/APRI, Fibroscan/ultrasonography and APRI/ultrasonography respectively.
Conclusion: Fibroscan appeared superior to APRI score and abdominal ultrasonography in diagnosis of liver fibrosis. Combined Fibroscan /ultrasonography performed better than other combinations for the prediction of significant hepatic fibrosis.

Open Access Original Research Article

Emergence of Antibiotic-Producing Microorganisms in Residential Versus Recreational Microenvironments

Yvon Woappi, Prashant Gabani, Om V. Singh

Microbiology Research Journal International, Page 280-294
DOI: 10.9734/BMRJ/2013/3205

Aims: To identify novel antibiotic-producing microbial strains with unprecedented pertinence. We hypothesize that site-specific soil samples will contain a variety of antibiotic-producing species (APS) with diverse specificity of molecular elements.
Place and Duration of Study: Laboratory of Microbiology, Division of Biological and Health Sciences, University of Pittsburgh, Bradford, PA-16701, USA, between August 2010 and May 2011.
Methodology: The environmental soil samples were collected from residential and recreational sites in Southern, PA, USA at longitude: -76 42 21.7116, latitude: 39 56 35.7252; approximately 201 meters above sea level. Over 70 natural antibiotic-producing soil bacteria were screened against 19 pathogenic microorganisms. Agar-plug assay was established to identify the antibiotics’ potency and pathogenic inhibitory index calculations were employed to measure the inhibitory potential of each isolate; 16S rRNA sequencing was used for microbial classification.
Results: A total of 71 microorganisms from residential soil demonstrated zones of inhibition (ZOI), followed by 9 organisms from recreational soil sample. A total of 15 bioactive strains demonstrated convincing growth inhibitory properties against 16 clinically relevant pathogens; 40% revealed pDNA presence, of which 67% exhibited stringent potencies against S. aureus. We observed a highly bioactive residential soil microbiota compared to recreational soil.
Conclusion: 16S rRNA sequence analysis corroborated several of the species belonging to Enterobacteriaceae, Xanthomonadaceae, and Bacillaceae. These findings may indicate a co-evolutionary biosynthesis of novel antibiotics driven by the increase of bioactive microbiota in residential environments.

Open Access Original Research Article

Microflora of Three Dehydrated Vegetables

Abosede. A. Akeredolu, Lawrence. O. Adebajo

Microbiology Research Journal International, Page 295-308
DOI: 10.9734/BMRJ/2013/3139

Consumption of fresh vegetables is very common in Nigeria. One of the methods of increasing the availability of vegetables all year round is drying. There is need to determine the microbial quality/safety of the dehydrated vegetables in order to determine the risk of food borne diseases. The microorganisms associated with three dehydrated vegetables (bitter leaf, bell pepper and okra) were isolated, identified and enumerated. The moisture contents of the vegetables were determined and the effects of varying temperature levels on growth of microbial isolates recovered from the samples were studied. A total of nine bacteria, namely: Staphylococcus aureus, Acinetobacter iwoffi, Corynebacterium sp., Bacillus pumilus, Micrococcus sp., Pseudomonas aeruginosa, Flavobacterium sp., Bacillus sp., Micrococcus kristianae and eleven fungi: Aspergillus niger, A. flavus, Aspergillus spp., Penicillium spp., Cladosporium sp., Fusarium spp. were isolated. The mean for total colony forming units (cfu/g) for bacteria were 2.1x107cfu/g, 6.1x105cfu/g, 2.2x106 cfu/g for bell pepper, bitter leaf and okra, respectively while the mean for total colony forming unit (cfu/g) for fungi were highest (1.3 x 106cfu/g) in bell pepper while bitter leaf recorded the least (7 x 103cfu/g) mean for total colony unit for fungi. The mean for percentage (%) moisture content ranged between 16.6-25.8%.The optimum growth was recorded for all the bacteria and fungi at 37ºC and 30ºC; nearly all the isolates had their growth retarded at 45ºC. The recovery of several harmful microorganisms in this study suggest the need for proper handling of vegetables during processing and storage to minimize microbial contamination in order to protect consumers’ health.

Open Access Original Research Article

Antifungal Effect of Surface Quick (HELVEMED) Solution on Candida Strains Isolated From Patients with Denture-Related Candidiasis

Jamileh Beigom Taheri, Somayyeh Azimi, Seyed Ali Razavi Rohani, Mohammad Reza Tarahhomi

Microbiology Research Journal International, Page 318-324
DOI: 10.9734/BMRJ/2013/3410

Aim: Surface Quick, an alcohol-based solution has been prepared by HELVEMED Company and various advantages have been mentioned for it; however all these claims have not been assessed scientifically. This study evaluated the antifungal efficacy of Surface Quick solution on candida albicans species obtained from the patients with denture-induced candidiasis.
Study Design: This is an experimental study for evaluation of antifungal effect of surface quick solution.
Place and Duration of Study: This study was done in Oral medicine and microbiology department of Shahid Beheshti University of Medical Sciences, Tehran, Iran in 2012.
Methodology: In this experimental in vitro trial, standard Candida albicans species (PTCC 5027) were obtained and the samples were prepared from 30 patients with denture-induced candidiasis using sterilized swab. Candida albicans species were grown in the sabouraud dextrose agar medium and the diameter of the inhibitory zone were calculated after exposing the medium to Surface Quick solution. The difference of inhibitory zone diameter between men and women was statistically analyzed by Student t test and the association of the individuals’ age and gender with inhibitory zone diameter was assessed by logistic regression analysis.
Results: The mean and standard deviation of the inhibitory zone diameter after exposing candida albicans species with Surface Quick solution was 10.13 mm and 3.33 mm. No significant differences were found between candida albicans species obtained from the male and female individuals. There was no significant association among participants’ age and gender with the inhibitory zone diameter.
Conclusion: According to the results, Surface Quick solution was effective against Candida albicans species obtained from candidacies patients even in the least concentration. Therefore this solution could be used for surface disinfection caused by candida albicans.

Open Access Original Research Article

Evaluation of in vitro Reduction of Hexavalent Chromium by Cell-Free Extract of Arthrobacter sp. SUK 1201

Satarupa Dey, A. K. Paul

Microbiology Research Journal International, Page 325-338
DOI: 10.9734/BMRJ/2013/3726

Aims: This study aims at to evaluate the hexavalent chromium [Cr(VI)] reduction potential of crude cell-free extracts of chromium resistant and reducing bacterium Arthrobacter sp. SUK 1201 and determination of optimum conditions for Cr(VI) reduction for possible bioremediation of Cr pollutants.
Place and Duration of Study: Chromium reduction studies with Arthrobacter sp. SUK 1201, was undertaken in the Microbiology Laboratory, Department of Botany, University of Calcutta, Kolkata during 2010-2012.
Methodology: Cell-free extract was prepared from freshly grown cell mass of Arthrobacter sp. SUK 1201 following the standard procedure. Cell mass suspended in Tris-HCl was sonicated (120 KHz for 30 min), centrifuged (12,000×g at 4ºC for 10 min) and the supernatant (S12) was used as the cell- free extract (CFE). Chromate reductase activity of the CFE was assayed colorimetrically using 1, 5-diphenylcarbazide as the complexing reagent.
Results: Chromate reductase activity of CFE of Arthrobacter sp. SUK 1201 was constitutive in nature and reduced Cr(VI) with decreasing efficiency as the concentration of Cr(VI) was increased. Its Km and Vmax were 263.45 μM Cr(VI) and 17.5 U mg-1 protein respectively. Reduction of Cr(VI) was optimal at pH 7 and 32ºC but was extremely thermolabile. NADH was the most suitable electron donor, and the chromate reduction was enhanced by Cu(II) and Fe(III), but inhibited by Hg(II). Among the different inhibitors tested, 2, 4-dinitrophenol (DNP) restored nearly 96.4% reductase activity, while carbonyl cyanide-m-chloro phenyl hydrazone (CCCP) was most inhibitory to the process.
Conclusion: It has been established that the Cr(VI) reduction potential of the cell-free extract of Arthrobacter sp. SUK 1201 is promising and could be exploited in the bioremediation of toxic hexavalent chromium.

Open Access Original Research Article

Disease Dynamics and Surveillance of Malaria in Malwa Region of Punjab and Evaluation of RDT Test

Renu Bansal, Neerja Jindal, Sandeep Sidhu

Microbiology Research Journal International, Page 339-345
DOI: 10.9734/BMRJ/2013/3344

Aim: Malaria remains an enormous public health problem. Regular and ongoing surveillance to detect changes in its trends to initiate the control measures is the need of the hour. The present study was undertaken to provide the malaria transmission dynamics using surveillance indicators through active and passive surveillance in district Faridkot. Usefulness of rapid malaria diagnostic test was also evaluated.
Methodology: This retrospective study extended over a period of two years (2010-2011). Thick and thin blood smears were prepared from suspected cases of malaria complaining of fever and headache for the last three days (i) of 2 CHC’s, 8 PHC’s and 68 sub centers as a part of active surveillance and (ii) those who visited GGS Medical College & Hospital and civil hospital Faridkot as a part of passive surveillance. Out of all the samples collected during the passive surveillance 995 samples collected at GGS Medical College and Hospital, Faridkot were also subjected to rapid diagnostic test (OptiMAL®).
Results: The annual blood examination rate (ABER) was 9.0 and 9.7 in 2010 and 2011 respectively. Annual parasite incidence (API) recorded was < 2 (0.5) in both the years and slide positive rate (SPR) was 0.5 and 0.05 in the two respective years of study. Significant gap in the rate of case detection of active and passive surveillance systems was observed with predominance of passive surveillance. More than 96% of cases were of P. vivax. RDT’s showed an excellent correlation with conventional microscopy.
Conclusion: Malaria (P. vivax) is a persistent problem in the Malwa region with variation in its transmission dynamics with in the year. P. vivax is the main species of malarial parasite in the Faridkot district with occasional cases of falciparum malaria. Prevention strategy should be targeted towards on the spot diagnosis by using RDT and hence prompt treatment. It could help to prevent spread of drug resistance and complicated malaria.

Open Access Original Research Article

Isolation of Microorganisms from Dairy Effluent

Vishakha Sukhadev Shivsharan, Minal Wani, M. B. Khetmalas

Microbiology Research Journal International, Page 346-354
DOI: 10.9734/BMRJ/2013/3445

Aim: Every Dairy industry has problems of effluent treatment. This can be revealed by effective treatment of the effluent. The effective treatment can be done by using microorganisms to stabilize the organic and inorganic load of the effluent. The aim of the present work is to study the dairy wastewater micro biota and to identify some new active strains which can bring about fast biodegradation of the organic compounds.
Study Design: Isolation and determination of bacteriological characteristics of the dairy effluents.
Methodology: Studies were carried out to isolate the microorganisms from collected effluent sample from the dairies under studies. Isolation of microorganisms was done by primary screening, Cultural Characterization, Biochemical characterization and Identified by using Bergey’s Manuals of Systematic Bacteriology.
Results: During 2011-2013 from two different districts of Maharashtra (India), dairy industry effluents were collected for the isolation of micro organisms. Effluent samples were collected as per Jacksch and piper method and primary screening was done and totally 7 Isolates were screened out. These isolates were characterized on nutrient agar at room temperature for 24 hrs. Isolates were observed for the cultural characters like size, shape, colour, margin, elevation, opacity and consistency and morphological characters like Gram nature, sporulation, shape and arrangement of cells, motility etc. and were recorded. Physio-biochemical characterization was followed by biochemical tests for enzymatic activities like catalase, oxidase, nitrate reduction, urease, caseinase etc and carbohydrates utilization tests for lactose, maltose, inositol, xylose etc. performed to check their ability for metabolization. On the basis of these characteristics, isolates were identified by using Bergey’s Manuals of Systematic Bacteriology. The identified Bacterial Isolates were of Genus Lactobacillus, Bacillus, Staphylococcus, Enterococcus, Listeria etc.
Conclusion: These Bacterial isolates have the ability to utilize the components like nitrate, starch, gelatin, sugars like sucrose, maltose, lactose etc. which was confirmed by the biochemical tests. Bacterial flora from the effluents can be identified and efficiently applied for the biological treatment of the dairy effluents.

Open Access Original Research Article

Induced; Thermal Influence on Morphology of Aspergillus carbonarius During Raw Starch Digesting Amylase Production

Onyetugo C. Amadi, Bartholomew N. Okolo

Microbiology Research Journal International, Page 355-367
DOI: 10.9734/BMRJ/2013/2815

Aim: To investigate increased thermal influence on morphology of Aspergillus carbonarius during RSDA production.
Place and Duration of Study: Microbial fermentation Unit, Department of Microbiology, Faculty of Biological sciences, University of Nigeria, between July 2009 and August 2010.
Methodology: In shake flask cultures thermal influence on A. carbonarius morphology and productivity investigated. Mycelial morphology was characterised by means of image analysis using as parameters, mean diameter, roughness, circularity and compactness of pellet. Thermal effect on amylase activity, total protein, biomass concentration and pH were also investigated.
Results: Shifting the temperature from 27ºC to 37ºC significantly affected the morphological parameters of the pellets, but RSDA activity was not altered. The interesting thing about the morphology is the shearing off of the hairy part of the pellet at an increased temperature and subsequent agglomeration. At 27ºC the RSDA activity increased steadily with an optimum activity of 293U/ml at 96h and subsequently decreased to 75U/ml by the end of the fermentation. At 37ºC a maximum activity of 291U/ml was achieved at 72h of fermentation but this decreased to 87U/ml at the end of fermentation. Higher biomass concentration and total protein were obtained at 37ºC. The pH dropped from an initial of 5.0 to 3.0 and 2.5 for 27ºC and 37ºC temperature conditions respectively.
Conclusion: Induced thermal increase resulted to changes in pellet morphology but raw starch digesting amylase activity was not altered.

Open Access Original Research Article

Isolation and Molecular Characterization of Lactic Acid Bacteria Isolated from Fresh Fruits and Vegetables Using Nested PCR Analysis

E. C. Emerenini, O. R. Afolabi, P. I. Okolie, A. K. Akintokun

Microbiology Research Journal International, Page 368-377
DOI: 10.9734/BMRJ/2013/2520

Aims: The study investigated the diversity and identities of Lactic Acid Bacteria (LAB) isolated from different fresh fruits and vegetables using Molecular Nested PCR analysis with the view of identifying LAB with anti-microbial potentials.
Study Design: Nested PCR approach was used in this study employing universal 16S rRNA gene primers in the first round PCR and LAB specific Primers in the second round PCR with the view of generating specific Nested PCR products for the LAB diversity present in the samples.
Place and Duration of Study: Biotechnology Centre of Federal University of Agriculture, Abeokuta, Ogun State, Nigeria, between January 2011 and February 2012.
Methodology: Forty Gram positive, catalase negative strains of LAB were isolated from fresh fruits and vegetables on Man Rogosa and Sharpe agar (Lab M) using streaking method. Standard molecular methods were used for DNA extraction (Norgen Biotek kit method, Canada), Polymerase Chain Reaction (PCR) Amplification, Electrophoresis, Purification and Sequencing of generated Nested PCR products (Macrogen Inc., USA).
Results: The partial sequences obtained were deposited in the database of National Centre for Biotechnology Information (NCBI). Isolates were identified based upon the sequences as Weissella cibaria (5 isolates, 27.78%), Weissella kimchi (5, 27.78%), Weissella paramensenteroides (3, 16.67%), Lactobacillus plantarum (2, 11.11%), Pediococcus pentosaceus (2, 11.11%) and Lactobacillus pentosus (1, 5.56%) from fresh vegetable; while Weissella cibaria (4, 18.18%), Weissella confusa (3, 13.64%), Leuconostoc paramensenteroides (1, 4.55%), Lactobacillus plantarum (8, 36.36%), Lactobacillus paraplantarum (1, 4.55%) and Lactobacillus pentosus (1, 4.55%) were identified from fresh fruits.
Conclusion: This study shows that potentially LAB can be quickly and holistically characterized by molecular methods to specie level by nested PCR analysis of the bacteria isolate genomic DNA using universal 16S rRNA primers and LAB specific primer.

Open Access Original Research Article

Enhancing Bio-Available Phosphorous in Soil Through Sulfur Oxidation by Thiobacilli

Irfan Ullah, Ghulam Jilani, Muhammad Irfan ul Haq, Anwar Khan

Microbiology Research Journal International, Page 378-392
DOI: 10.9734/BMRJ/2013/4063

Aims: The objectives were to evaluate the phosphate solubilization efficiency of different Thiobacilli strains and to find out the best combination of sulfur and Thiobacilli for enhancing bio-available P in soil.
Study Design: An experimental study.
Place and Duration of Study: Microbiology and Soil Fertility Labs, Department of Soil Science and Soil and Water Conservation, Pir Mehr Ali Shah Arid Agriculture University, Rawalpindi, Pakistan and Microbiology and Soil Chemistry Labs, Auriga Research Center, Lahore, Pakistan, between May 2011 and November 2012.
Methodology: Fifty Thiobacilli strains were isolated from ten different ecologies. Then an incubation study of soil was performed wherein the most efficient four Thiobacilli strains were inoculated in combination with three different levels of elemental sulfur to determine pH, water soluble sulfur, sequential P fractions and bio-available phosphorous contents in the incubated soil.
Results: All the four Thiobacillus strains (IW16, SW2, IW1 and IW14) dropped pH of the incubated soil along with three doses of S° (50, 75 and 100 kg ha-1). However, Thiobacillus strains IW16 and SW2 reduced soil pH quite sharply from 7.90 to 7.12 (net reduction of 0.78 points) and 7.28 (net reduction of 0.62 points) respectively where inoculated with S° @ 100 kg ha-1. The best P solubilizer was Thiobacillus strain IW16 and the best dose of S° was @ 100 kg ha-1 and their combination enhanced maximum quantity of P (22.26 mg kg-1) in the soil by solubilizing already present insoluble calcium bounded P fractions like octa-calcium phosphate (Ca8-P) and apatite (Ca10-P).
Conclusion: The present study suggests the use of Thiobacilli along with elemental sulfur for the dissolution and enhancement of bio-available P in alkaline and calcareous soils.

Open Access Original Research Article

Parasitological Evaluation of Domestic Water Sources in a Rural Community in Nigeria

Solomon Chuwang Chollom, Michael Unata Iduh, Bitrus Joseph Gyang, Micheal Abbah Idoko, Aloysius Ujah, Godwin Ojonugwa Agada, Joan Peter, Yomi Richard Akele, Joseph Ameh Okwori

Microbiology Research Journal International, Page 393-399
DOI: 10.9734/BMRJ/2013/3513

Aim: To evaluate the level of safety of water sources in a rural settlement in Nigeria with reference to parasitic infections and to make appropriate recommendations to the government and the community dwellers.
Study Design: Investigative study.
Place and Duration of Study: Samples were collected in Heipang, Barkin Ladi Local Government Area of Plateau State, Nigeria between October-December, 2012. They were processed at the General Laboratory of National Veterinary Research Institute, Vom, Nigeria.
Methodology: 100 water samples were collected from domestic water sources. 10 of the samples were from streams, 60 from ponds, 20 from wells and 10 were from bore holes. Samples were investigated for presence of parasites using standard World Health Organisation approved laboratory techniques. Each sample was subjected to macroscopy, filtration, centrifugation and microscopy.
Results: It revealed that 59 out of 100 water sources investigated had parasitic infestation. Ponds had the highest degree of parasitic contamination (78.3%), streams followed closely with 50%, while wells and bore holes had 35% and 0% in that order. Helminths were the leading parasitic genera encountered with Ascaris species accounting for 33.9% of the parasites. Hookworm was the second most common helminth with the prevalence of 20.3%. Strongyloides species accounted for a paltry prevalence of 3.4%. Protozoan cysts of Balanditium coli and Entamoeba histolytica accounted for 18.6% of parasites each.
Conclusion: These findings clearly show that most water sources in the study area constitute grave epidemiological threat to public health. Inhabitants of such communities should boil or treat their water before consumption while government authorities should provide safe drinking water to the rural dwellers.

Open Access Original Research Article

Prevalence of Human Papilloma Virus DNA in HIV Positive Women in Lagos University Teaching Hospital (LUTH) Lagos, Nigeria

I. G. Nweke, A. A. F. Banjo, F. B. Abdulkareem, V. U. Nwadike

Microbiology Research Journal International, Page 400-413
DOI: 10.9734/BMRJ/2013/4151

Introduction: HIV-infected women have a high prevalence of Human Papilloma virus (HPV) infection and are more likely to be infected with high risk genotypes with the potential of progressing to cervical cancer. There is paucity of data regarding the prevalence of sexually transmitted HPV infection among HIV positive women in Nigeria.
Aims: The objective of this cross-sectional prospective study was to determine the prevalence of high risk HPV among HIV positive and negative women in LUTH, Lagos, Nigeria and to relate HPV genotypes in the study population to commercially available HPV vaccine types that would be or not be appropriate for implementation of vaccination programs in Lagos State.
Place and Duration of Study: AIDS Prevention Initiative In Nigeria (APIN) clinic as well as the Gynecologic outpatient clinic of LUTH, Lagos between August 2011 and August 2012.
Methodology: A combination of PCR and flow through hybridization method was used in the genotyping of HPV from samples obtained from 98 HIV positive and 97 HIV negative women. Data was analyzed using Epi info 3.5.6. Non parametric variables were compared with chi-square or Fisher exact test as appropriate. The differences in mean for parametric variables were compared using student T test. P value <0.05 were considered significant.
Results: The prevalence of HPV among HIV positive women was 44.9% while the prevalence of high risk types was 37.5%. The commonest high risk types seen were types 31, 52, 53 and 35. The prevalence of HPV among the HIV negative women was 11%. The commonest high risk types seen were types 18, 16, 52 and 56.
Conclusion: In view of the high prevalence and diversity of HPV genotypes among the HIV positive women, adequate screening protocols should be put in place for screening this category of women. Studies should also be carried out to determine the efficacy of existent HPV vaccines on this group of patients.

Open Access Original Research Article

Prevalence of papC and usp Virulence Factors in Uropathogenic Escherichia coli Causing Asymptomatic Urinary Tract Infections in Adolescents

Samuel Nkansah Darko, Kwabena Nsiah, Peter Twumasi

Microbiology Research Journal International, Page 423-430
DOI: 10.9734/BMRJ/2013/3187

Aims: To determine the prevalence of two virulence genes associated with uropathogenic Escherichia coli; papC gene of the P fimbriae for adherence to uro-epithelial cells and usp (uropathogen-specific protein) gene, a Vibrio cholerae toxin gene homologue.
Study Design: Cross sectional.
Place and Duration of Study: Department of Biochemistry and Biotechnology and the Clinical Analysis Laboratory, Kwame Nkrumah University of Science and Technology, Kumasi, between October 2011 and February 2012.
Methodology: Escherichia coli isolates (n= 149) from an adolescent population of ages 13-18 years (from a total sampled population of 85 males and 64 females) were screened for papC and usp, using specific primers for the two genes in polymerase chain reactions.
Results: The usp gene was the most prevalent (72.48%), followed by papC (51.00%) and papC+usp (24.16%). Significant difference (P = .002) was observed between papC and usp and also papC and papC+usp (P < .0001). usp Gene prevalence was also significantly different from that of papC+usp (P < .0001).
Conclusion: This study suggests that a higher proportion of strains of uropathogenic Escherichia coli implicated in UTI in the studied population possess the usp gene whose protein product potentially serves to reduce competing microbes in the urinary tract.

Open Access Original Research Article

Prevalence and Antimicrobial Susceptibility of Salmonella Species Associated with Childhood Acute Gastroenteritis in Federal Capital Territory Abuja, Nigeria

Ifeanyi Casmir Ifeanyichukwu Cajetan, Bassey Enya Bassey, Ikeneche Nkiruka Florence, Isu Rosemary Nnennaya, Akpa Alexander Casmir

Microbiology Research Journal International, Page 431-439
DOI: 10.9734/BMRJ/2013/2379

Aim: The aim of this study was to isolate and characterize Salmonella strains associated with childhood acute gastroenteritis in Nigeria; as well as to evaluate the resistant patterns of the strains to the commonly used antimicrobials agents.
Study Design: Children ≤ 5years having diarrhoea characterized by the occurrence of three or more loose or watery stool or at least one bloody loose stool in a 24-hour period were enrolled in the study.
Methods: The study was conducted between July and December 2008. Samples were pre-enriched in buffered peptone water followed by selective enrichment using selenite cysteine and Rapaport-Vassilidis broths. Isolation and identification was made by inoculating the selectively enriched sample on to Xylose Lysine Deoxycholate agar followed by confirmation of presumptive colonies using different biochemical tests. The CLSI, 2006 method was used for antimicrobial susceptibility testing.
Results: In all the 400 tested samples, 9 (2.3%) were positive for Salmonella isolates. Results showed that the children aged 0-5 months had the highest Salmonella infection rate of 5 (4.1%), followed by 13-24 months 4 (3.5%), while Salmonella infection was not present in the age groups of 25-36 months, 37-48 months, and 49-60 months.
The highest (3.2%) Salmonella infection rate was seen among children on solid food followed by those on breast milk (2.5%), while those on a combination of breast milk and formula had no detectable level of Salmonella infection.
The study recorded various degrees of resistance to four antimicrobials as observed in amoxicillin, cephalexin, and cefuroxime (55.6%) each, while resistance was observed in 77.8% of the isolates against amoxycillin-clavulanic acid. All isolates were susceptible to ciprofloxacin, nalidixic acid, and Ceftriaxone.
Conclusion: The study reports Salmonella species as a potential pathogen isolated from stool samples of children with acute gastroenteritis. The overall resistance level of the isolates to amoxycillin-clavulanic was highest followed by resistance to amoxicillin, cephalexin, cefuroxime giving a cause for concern.

Open Access Original Research Article

Evaluation of Phospholipase Activity in Biofilm Forming Candida Species Isolated from Intensive Care Unit Patients

Sachin Deorukhkar, Santosh Saini

Microbiology Research Journal International, Page 440-447
DOI: 10.9734/BMRJ/2013/4359

Aims: To evaluate phospholipase activity in biofilm forming Candida spp. isolated from patients admitted in intensive care unit of rural tertiary care hospital.
Study Design: A total of 135 biofilm forming Candida spp. isolated from various clinical specimens of patients admitted in ICU were included in the study.
Place and Duration of Study: Department of Microbiology, Pravara Institute of Medical Science’s Rural Medical College India, between January 2010 and December 2012.
Methodology: The Candida isolates were identified upto species level by conventional standard mycological techniques. The biofilm formation was assessed by inoculating the isolates in conical polystyrene test tube containing Sabouraud’s dextrose broth supplemented with glucose. Phospholipase activity of biofilm forming Candida isolates was detected by using egg yolk agar.
Results: Out of 135 biofilm forming Candida spp. included in the study, 60 (44.4%) isolates were C. albicans. Among non-albicans Candida (NAC) spp. C. tropicalis was the major isolate followed by C. glabrata and C. parapsilosis. Phospholipase production was seen in 85 (62.9%) isolates. A total 49 (81.6%) isolates of C. albicans showed phospholipase activity. Among NAC spp. maximum phospholipase activity was seen in C. tropicalis and C. glabrata.
Conclusion: Biofilm formed by the Candida spp. tend to be more resistant to antifungal drugs. Though C. albicans the most common species associated with Candida biofilms, the emergence of NAC spp. is of concern. NAC spp. shows varying degree of resistance either intrinsic or acquired or both to commonly used antifungal drugs. The isolation of NAC spp. from clinical specimens is no longer overlooked as these organisms are emerging pathogens. The virulence factors like biofilm formation and phospholipase activity is also noted in NAC spp. The study of these virulence factors would help in understanding the pathogenic role of NAC spp.

Open Access Review Article

Cellulase Production by Bacteria: A Review

Sangrila Sadhu, Tushar Kanti Maiti

Microbiology Research Journal International, Page 235-258
DOI: 10.9734/BMRJ/2013/2367

Cellulose is an abundant natural biopolymer on earth and most dominating Agricultural waste. This cellulosic biomass is a renewable and abundant resource with great potential for bioconversion to value-added bioproducts. It can be degraded by cellulase produced by cellulolytic bacteria. This enzyme has various industrial applications and now considered as major group of industrial enzyme. The review discusses application of cellulase, classification of cellulase, quantification of cellulase, the types of cellulolytic bacteria and their screening. It describes the current knowledge of cellulase production by submerged fermentation and solid state fermentation, properties of cellulase and cloning and expression of cellulase gene. The biotechnological aspect of cellulase research and their future prospects are also discussed.

Open Access Review Article

Quorum sensing and interspecies interactions in Stenotrophomonas maltophilia

R. AboZahra

Microbiology Research Journal International, Page 414-422
DOI: 10.9734/BMRJ/2013/4291

Stenotrophomonas maltophilia is a gram-negative bacterium that is widespread in the environment and that has become important in the last years as an emerging opportunistic pathogen. Quorum sensing (QS) is a bacterial cell–cell communication process that involves the production, detection, and response to extracellular signalling molecules called autoinducers.
S. maltophilia has a diffusible signal factor (DSF) that controls cell–cell communication and many functions such as motility, extracellular protease production and microcolonies formation in artificial sputum medium. This DSF signalling mediates also interspecies interactions between S. maltophilia and Pseudomonas aeruginosa such as susceptibility to polymixin and its influence on biofilm formation.
The traditional approach for the treatment of infectious diseases is to kill or inhibit the growth of bacteria using antibiotics. In response to the rise in antibiotic resistance, the development and use of QS inhibition based drugs to attenuate bacterial pathogenicity is now highly required in the microbiological and clinical fields.