Open Access Case study

Co-Infection of Hepatitis B Virus and Hepatitis Delta Virus in Yaounde-Cameroon

Judith Ndongo Torimiro, Gwladys Chavely Monamele, Mathurin Pierre Kowo, Désiré Takou, Guy-Bertrand Pouokam, Joseph Fokam, Oudou Njoya

Microbiology Research Journal International, Page 1381-1391
DOI: 10.9734/BMRJ/2014/6894

Aims: To determine the seroprevalence of HDV as well as the virological and clinical characteristics of HBV mono-infected and HBV/HDV co-infected patients.

Study Design: The few studies on HDV in Cameroon have reported a high prevalence of this viral infection. This is a first step in describing the virological and clinical profile of HBV mono-infected and of HBV/HDV co-infected patients.

Place and Duration of Study: Blood collection was carried out in the Gastroenterology Unit of the Yaounde University Hospital Centre, Yaounde General Hospital and “Centre Médical la Cathédrale”, from August 2012 to May 2013.

Methodology: We included into this study treatment-naïve HBV-infected patients from Yaounde irrespective of age and gender free of HIV and HCV infection. Blood samples were collected from each patient for laboratory analysis. Detection of HDV antibodies (Diasorin, Germany) was performed by ELISA and viral load for HBV and HDV was determined using real-time PCR (Abbott Molecular Diagnostics). Patients were classified clinically into low replicative hepatitis, immune tolerance and chronic active hepatitis. Moreover, ultrasound and liver histological data were collected.

Results: The population comprised 128 chronic HBV-infected patients of which 77 (60.16%) were male and 51 (39.84%) were female. We found 29 HDV-positive patients representing 22.66% of the population. In the HBV/HDV co-infected group, the mean viral load for HBV was significantly low compared to patients with HBV mono-infection (P = .01). These patients also presented with higher liver cytolysis compared to HBV mono-infected patients (P<.001). Chronic active hepatitis was significantly more prevalent in HBV/HDV co-infected patients (68.96%) compared to HBV mono-infected patients (20.20%). 

Conclusion: We found that HBV/HDV co-infection results in suppression of HBV replication and such patients show broader sequelae of liver disease. The prevalence of HBV and HDV co-infection is high in this population. Routine screening of HBV-positive individuals for HDV should be implemented in the health services nationwide.

Open Access Original Research Article

Trans Sialidase Genes Allow Clustering of TcI Trypanosoma cruzi Mexican Isolates

V. M. Monteón, R. López, A. Ramos-Ligonio, K. Acosta-Viana

Microbiology Research Journal International, Page 1299-1310
DOI: 10.9734/BMRJ/2014/11060

Aims: Polymorphism of the trans sialidase (TS) family of genes is common in Trypanosoma cruzi. Our goal was to cluster Mexican TcI DTU (Discrete Typing Unit) using a set of primers specific for TS genes.

Methodology: The DNA of 12 Mexican T. cruzi stocks (TcI) and reference strain were amplified using the CRP-1 primer, which anneals to the conserved 5' ends of CRP (Complement Regulatory Protein), TS, and FL-160 genes, and the CRP-2 primer, which anneals to conserved region within the GPI (Glycosil  Phosphatidil  Inositol) anchor sequence. Amplicons were analysed using PCR-SSCP (Single Strand Conformation Polymorphims) followed by construction of a nominal matrix data (presence/absence bands) to calculated the Jaccard and Dice similarity coefficient, and clustering with UPGMA.

Results: Mexican TcI stocks produced a common pattern of amplification products and cluster in a separate group to CL-Brener strain (TcVI). The PCR-SSCP revealed that within the TcI Mexican stocks there were a complex pattern, but T. cruzi from the Yucatan peninsula clustered in special and separate group.

Conclusion: The CRP-1 and CRP-2 primers were helpful for the analysis of genetic traits in T. cruzi DTU I and revealed the existence of special group in Yucatan Mexico.

Open Access Original Research Article

Antibiotic Susceptibility and Intestinal Epithelial Cell Adhesion Pattern of Homo Fermenting Lactic Acid Bacteria (LAB) Isolated from Kunu-Zaki, a Spontaneously Fermenting Nigerian Cereal Beverage

S. O. Oluwajoba, F. A. Akinyosoye, V. O. Oyetayo

Microbiology Research Journal International, Page 1311-1324
DOI: 10.9734/BMRJ/2014/10772

Aim: To determine the antibiotic reaction and adhesion pattern of antimicrobial homo-fermenting LAB strains in the fermenting slurries of kunu zaki.

Study Design: ANOVA. Inhibition of indicator lawn used ≥10mm inhibition as antibiotic susceptible. Adhesion was measured by staining and quantifying recorded as percentage and index values.

Place and Duration of Study: Department of Microbiology, Federal University of Technology Akure and Biotechnology Unit, Federal Institute of Industrial Research, Oshodi, Nigeria between June, 2012 and December, 2012.

Methodology: Kunu-zaki drinks were produced using spontaneously fermenting cereal grains of Digitaria exilis (acha), Sorghum bicolour (sorghum) and Pennisetum americanum (millet) in composite and non-composite proportions. LAB isolates were obtained on MRS agar. Homo-fermenting isolates were identified to species level using the API 50 CHL test kit. Antibiotic sensitivity testing on the identified isolates followed the modified standard Kirby-Bauer procedure on MRS agar (pH 7.4) using the disc diffusion technique with selected antibiotics. For quality control of the antibiotics, sensitive reference strains            S. aureus ATCC 25923 and E. coli ATCC 25922 obtained from the Nigeria Institute of Medical Research were used. Adhesion and antimicrobial properties were determined using standard method.

Results: Antimicrobial substances produced by the eight LAB isolates inhibited the growth of four selected human pathogens in vitro. All eight LAB isolates were resistant to amoxicillin, gentamycin and ciprofloxacin. L. plantarum126, L. paracasei subsp paracasei339 and Pediococcus damnosus32 were resistant to erythromycin whilst all others were susceptible. L. plantarum126 and L. paracasei subsp paracasei339 were resistant to all antibiotics tested. All LAB isolates demonstrated high in-vitro intestinal epithelial cell adhesion potential.

Conclusion: LAB antimicrobial activity may not be affected if kunu zaki is consumed simultaneously with these antibiotic therapies. However, if these LAB strains are intended for use as kunu-zaki starter cultures, it is important that they should be further carefully examined for inability to transfer antibiotic resistance genes to food pathogens

Open Access Original Research Article

Seroprevalence of Anti-Human Metapneumovirus Antibodies in Hospitalized Children in Suleimani City/Iraq

Tariq A. G. Aziz, Nabil Salmo, Ali H. Bayati

Microbiology Research Journal International, Page 1325-1334
DOI: 10.9734/BMRJ/2014/11658

Background: Human Metapneumovirus (hMPV) is an important respiratory viral pathogen among children, and it is one of the causes of pediatric hospital admissions due to acute respiratory tract infections.

Objective: This study was done to predict the seroprevalence of anti-hMPV antibodies among hospitalized children presenting with acute respiratory tract infections in Suleimani Governorate, Kurdistan Region/Iraq.

Place and Duration: This study was done at the department of microbiology, school of medicine, suleimani University, between April 2011 and March 2012.

Methods: Indirect immunofluorescent assay (IIFA) was performed to detect serum anti-hMPV antibodies (IgM and IgG antibodies) from three hundred hospitalized children less than 5 years old with acute respiratory tract infections.

Results: IgM anti-hMPV antibodies were positive in thirty six (12%) out of three hundred children. The highest seroprevalence was found in the age group <1 year old, while the lowest in the age group 4 to <5 years old. No significant gender difference was found among seropositive children. The IgM anti – hMPV seropositive children were suffering from pneumonia, bronchiolitis, or other less severe acute respiratory tract infections like acute bronchitis and croup in frequencies of sixteen (44%), 10 (28%), and 10 (28%). The IgG anti-hMPV antibodies were positive in two hundred and twenty five (75%) out of the three hundred children, and there was a gradual increase in percentage of seropositivity with increasing age.   

Conclusion: hMPV is an important viral respiratory pathogen among hospitalized children in Suleimani Governorate/Kurdistan/Iraq, and most of the children had experienced hMPV infection by the age of five years.

Open Access Original Research Article

Clindamycin Resistance among Staphylococcus Aureus Isolated at Mbarara Regional Referral Hospital, in South Western Uganda

Bashir Mwambi, Jacob Iramiot, Freddie Bwanga, Marthae Nakaye, Herbert Itabangi, Joel Bazira

Microbiology Research Journal International, Page 1335-1344
DOI: 10.9734/BMRJ/2014/10572

Aims: The study was conducted to determine the prevalence of Clindamycin (CL) resistance and antimicrobial susceptibility among clinical isolates of Staphylococcus aureus (S. aureus) from Mbarara Regional Referral Hospital (MRRH) in Southwestern Uganda.

Study Design: Laboratory based cross sectional study.

Place and Duration of the Study: The study was conducted at the Microbiology department of Mbarara Regional referral hospital between November 2012 and December 2013.

Methodology: In our study, we recruited 300 S. aureus isolates that were stored in the laboratory and were obtained from different clinical samples. The isolates were tested for antimicrobial susceptibility by phenotypic methods and for the genotypic expression of Macrolide Lincosamide StreptograminB (MLSB) resistance genes (ermA, ermB, ermC, and msrA). The D-test was also performed.

Results: Phenotypically, a total of 109 (36%) S. aureus isolates were resistant to CL, of which 9 (3%) were constitutively resistant while 100 (33.3%) were inducibly resistant. Genotypicaly, 134/300 (44.7%) isolates possessed at least one of the MLSB resistance genes. 23/300 (7.7%) tested positive for ermB, 98/300 (32.7%) tested positive for the ermC and 43/300 (14.3%) tested positive for the msrA genes with none possessing the ermA gene. Isolates were highly resistant to Sulfamethoxazole/trimethoprim, Erythromycin and Oxacillin with moderate resistance to Vancomycin and Imipenem and least resistance to Linezolid

Conclusion: S. aureus resistance to CL was high in this set up. There was also high resistance to Sulfamethoxazole/trimethoprim, Erythromycin and Oxacillin but low resistance to Linezolid.

Open Access Original Research Article

Molecular Characterization of Bacteria Isolates from Farm-Raised Catfish Clarias gariepinus (Burchell, 1822)

A. A. Akinyemi, O. O. Oyelakin

Microbiology Research Journal International, Page 1345-1352
DOI: 10.9734/BMRJ/2014/11996

Selected bacterial isolates from skin, gut and gills of Clarias gariepinus were collected from five fish farms at Ijebu Ode. The isolates were assessed using 16S rRNA gene sequencing method to identify them and to construct the phylogenetic relationship. A total of 10 isolates were selected, their colonial morphology determined, thereafter the DNA of the isolates were prepared using CTAB method, PCR amplification of 16S ribosomal RNA gene of isolates was carried out using universal primer for bacteria,  purification of the PCR product using ethanol precipitation, thereafter sequenced using an automated DNA sequencer. These sequence data were compared with other gene sequences in GenBank database (NCBI) using a BLAST search to find closely related sequences. 80% of the isolates belonged to different species of Pseudomonas, sharing 92% to 96% 16S ribosomal RNA identity with the respective type-strain, whereas the remaining 2 isolates belonged to Pediococcus acidilactici and Lysinibacillus fusiformis with 96% 16S ribosomal RNA homology.

Open Access Original Research Article

Antimicrobial Activity against Some Saprophytic and Pathogenic Microorganisms of Bacillus species Strains Isolated from Natural Spring Waters in Bulgaria

Yulian Tumbarski, Nedyalka Valcheva-Zhekova, Irina Koleva, Zapryana Denkova

Microbiology Research Journal International, Page 1353-1369
DOI: 10.9734/BMRJ/2014/11408

It is well known that many members of genus Bacillus possess an antimicrobial activity against a variety of microorganisms. In this study we present a primary screening for antimicrobial activity against some saprophytic and pathogenic microorganisms of nineteen strains identified as Bacillus cereus, Bacillus thuringiensis and Bacillus subtilis, isolated from eight natural thermal springs in two districts - Haskovo and Stara Zagora, Bulgaria. The inhibitory activities of the Bacillus sp. strains were determined by agar-well diffusion method. Test microorganisms were preliminarily included into the agar medium, whereas the Bacillus sp. strains were added to the wells. After 48 hours of incubation, the antimicrobial effects were determined by measuring the diameter of zones of inhibition around the wells. Most of Bacillus sp. strains showed high antimicrobial activitiy against the molds Penicillium sp., Aspergillus oryzae, Aspergillus niger, Aspergillus awamori, Fusarium moliniforme and Rhizopus sp. Inhibitory activity against the mold Mucor sp. and bacterium Enterococcus faecalis ranged between low and moderate. Two of the strains (Bacillus cereus 52/GI1 and Bacillus thuringiensis 56/H3) possessed moderate antimicrobial activity against bacterium Pseudomonas aeruginosa and yeasts Saccharomyces cerevisiae. One strain (Bacillus subtilis 47/YA1) had high activity against the mold Mucormucedo; one strain (Bacillus cereus 36/GI6) showed moderate activity against yeasts Candida utilis, while bacterium Escherichia coli was not inhibited at all.

Open Access Original Research Article

Preliminary Detection of Bacillus species in Commercial Honey

Obakpororo E. Agbagwa, Tosanwumi V. Otokunefor, Nnenna Frank-Peterside

Microbiology Research Journal International, Page 1370-1380
DOI: 10.9734/BMRJ/2014/7217

Aim: This research is aimed at detecting the presence of Bacillus species in honey using morphological characteristics, biochemical tests and preliminary molecular studies.

Place and Duration of Study: The study was carried out in the department of Molecular Biology, Institute of Medical Research Yaba Lagos, Nigeria. The study was carried out from   June to July 2012.

Methodology: A total of 33 honey samples were used for this study, twenty- eight of the honey samples were of local origin while 5 were of international origin. Twenty-eight commercial honey samples were obtained from the six geographical regions in Nigeria from commercial retailers. The five foreign honey samples were obtained from the supermarkets namely: Friz fruit, Blossom, Forever, Aloe Vera and Rose honey, all of international origin. The honey samples were inoculated into sterile agar, blood and tryptone soy plates using the spread plate technique. Isolates obtained were purified and subjected to morphological tests, biochemical tests and further identification using polymerase chain reaction.

Results: All the honey samples had microbial growth in them, higher counts were observed in the commercial honeys from retailers than the foreign honey samples. Forty isolates suspected to be Bacillus from biochemical tests were subjected to PCR, 14 from the 40 were confirmed to be Bacillus spp.

Conclusion: Microorganisms in honey cannot be identified fully with morphological and biochemical examinations alone, but combine use of morphological, biochemical tests and PCR technique is more accurate and reliable method of identification.

Open Access Original Research Article

Rhodotorula glutinis and Its Two Mutants Ameliorate Hepato-Renal Dysfunction Induced by Ochratoxin A on Rats

Wafaa Haggag, Enayat A. Omara, Somaia A. Nada, Hamdy Abou Eleid, Hassan A. Amra

Microbiology Research Journal International, Page 1392-1408
DOI: 10.9734/BMRJ/2014/10805

Aim: This study was designed to investigate the possible curative effect of Rhodotorula glutinis (R. glutinis) and its two mutants (Col-1R1 and Col-1R3) against hepatorenal toxicity induced by ochratoxin (OA) in rat.

Methods: The strains of yeast Col- 1R1 and Col- 1R3 have been genetically improved and isolated from R. glutinis after colchicines treatment. OA was produced and determined from Aspergillus ochracus isolate from Egyptian corn. Experimental design: Five groups of rats were treated as follows: group 1, was the control group orally given 4 ml / Kg 0.1 M NaCOH3; group 2 treated with OA (1.7 mg /Kg).Groups 3, 4 and 5 orally administered the R. glutinis and its two mutants (50 X106 colony forming unit (cfu) / 10 ml saline / kg body weight) prior 1hr of OA -treatment for 15 successive days.

Results: The studied autoploidy strains showed significant increase in caratenoids level, protease, β-1, 3-glucanase and chitinase activities when compared with the parental strain.

Biochemical results revealed that OA significantly decreased serum total antioxidant capacity (TAC) and it caused elevation inserum  transaminases (AST, ALT), creatinine, uric acid, nitric oxide (NO), tumor necrosis factor alpha (TNF-α) and carcinoembryonic antigen (CEA) (P <0.05) as compared with the control group. The three tested yeasts significantly decreased the elevated values toward the normal levels and improved the pathological feature in liver and kidney tissues. Moreover, R. glutinis and the two mutants significantly reduced hepatorenal damaged arias, increased optical density of DNA and alleviated ochratoxin A-induced caspase-3 activation. The resultant effect of the two mutant strains had more powerful effect more than the wild strainto ameliorate hepatorenal dysfunction in ochratoxicosis-rat.

Conclusion: Col-1R3 was more effective than Col-1R1 may be due to its higher contents of carotenoids, glucane and chitine, which act as antioxidants.

Open Access Original Research Article

Microbiological Quality of Dairy Cattle Products

Okeke Kingsley Susan, Abdullahi Isa Obansa, Makun Hussaini Anthony

Microbiology Research Journal International, Page 1409-1417
DOI: 10.9734/BMRJ/2014/11112

Aims: To determine the chemical properties (pH, titratable acidity) and microbiological qualities of fresh cow milk and traditional cultured skimmed (defatted) milk (nono) and full fat or partially skimmed cultured milk (kindirmo) in Bida local government area of Niger State, Nigeria.

Study Design: To assess the microbial load of dairy cattle products.

Place and Duration of Study: Samples were collected from local farmers in Madobia and Project quarters in Bida Local Government, Nigeria. Analyze at laboratories of Microbiology Department of Federal University of Technology, Minna and Federal Polytechnic, Bida between September 2011 and December 2012.

Methodology: Ninety samples of fresh milk, nono and kindirmo obtained from two areas in Bida Local Government Area were analyzed to determine their pH, titratable acidity, microbial properties (Total viable count, Fungal count, Staphylococcal count, Coliform count) and antibiogram of pathogenic organisms isolated from the samples. Results obtained were subjected to statistical analysis.

Results: The results obtained showed that the pH of nono was more acidic than other milk products. The total viable count ranged log106.02-6.36cfu/ml, coliform count log10 6.02-6.57cfu/ml, staphylococcal count log10 6.10-6.57cfu/ml; fungal count log10 4.49-5.10cfu/ml respectively. The microorganism isolated included Bacillus subtilis, Enterococcus faecalis, Staphylococcus aureus, Aspergillus flavus and Aspergillus niger. Notably S. aureus and A. flavus were frequently isolated (60.1% and 44% respectively). The antibiogram of pathogenic organisms isolated from the dairy cattle products showed that E. faecalis and S. aureus were sensitive to gentamicin (10µg) and streptomycin (30µg).

Conclusion:  The growth of these pathogenic organisms in local dairy cattle products is a reflection of poor sanitary practices in the production of fresh milk and its products. This high microbial load in cow milk and its product may pose a great public health concern and therefore calls for public awareness campaign.

Open Access Original Research Article

Purification, Biochemical Characterization and Applications of Pleurotus ostreatus ARC280 Laccase

Abdelmageed M. Othman, Ali M. Elshafei, Mohamed M. Hassan, Bakry M. Haroun, Maysa A. Elsayed, Ayman A. Farrag

Microbiology Research Journal International, Page 1418-1439
DOI: 10.9734/BMRJ/2014/11218

Aims: To purify, characterize, and apply the laccase produced by submerged fermentation using an edible mushroom Pleurotus ostreatus ARC280.

Study Design:  Laccase purification and characterization were designed using the most recent approaches and statistical studies of triplicate results values.

Place and Duration of Study: Department of Microbial Chemistry, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt, between May 2011 and January 2013.

Methodology: P. ostreatus ARC280 laccase was purified using ammonium sulfate precipitation (40-80%), followed by gel filtration using Sephadex G100 column chromatography. The resulted pure laccase was analyzed on SDS-PAGE (12%). Laccase activity parameters such as temperature, pH, stability, metal ions and kinetic constants were studied. Laccase was applied to reduce four tumor cell lines growth and as antibacterial and antifungal agent.

Results: P. ostreatus ARC280 laccase was purified using ammonium sulphate followed by Sephadex G-100 chromatographic column by about 148 purification fold with Mr of 85kDa. Optimum P. ostreatus ARC280 purified laccase activity was recorded at 50ºC and at pH 6.0, 3.0, 4.5 for Syringaldazine (SGZ), 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic) acid (ABTS) and 2, 6-dimethoxyphenol (DMP) as substrates, respectively. The purified enzyme was more stable in alkaline pH range and retained about 37.42, 73.51, 85.65, 87.7, 88.49, 93.65, 92.86 and 100.0 % of the initial activity after 5hrs of incubation at pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0, respectively. Hg2+ caused complete inhibition at all tested concentrations; however Mn2+ (2.5x10-3M) caused laccase activation by about 190 and 330% after 1 and 24 hrs, respectively. Km and Vmax were calculated and found to be 0.074, 2.857 and 0.476 µM and 1.563, 2.500 and 2.632 µmol min-1 for SGZ, DMP and ABTS, respectively. The purified enzyme has the ability to reduce four tested cell lines growth in vitro with percentage reduction of 16.8, 23.4, 15.2 and 23.4% for HePG2, HCT116, A549 and MCF7, respectively. On the other hand, the enzyme was found to have antibacterial and antifungal activities against Escherichia coli and Candida albicans respectively.

Conclusion: This enzyme seems to be a prospective enzyme for further biotechnological exploitation such as anticancer and antimicrobial activity applications.

Open Access Original Research Article

Biodegradation of Palm Oil Mill Effluent

P. N Ibegbulam-Njoku, O. K. Achi

Microbiology Research Journal International, Page 1440-1450
DOI: 10.9734/BMRJ/2014/12008

Aims: The aim of this study was to investigate the biodegradation capacity of selected indigenous fungal isolates and optimization of their degradation ability using various environmental factors such as pH, incubation temperature, nutrient concentration and inoculums size in reducing pollution effect of palm oil mill effluent (POME) in the environment.

Place and Duration of Study: Two fungal isolates Candida rugosa and Geotrichum        candidum used in this work were previously isolated from POME sample collected from Starline palm oil mill industries, Umukalika, Obingwa LGA, Abia state Nigeria in previous work of  authors. The study was carried out from March to August, 2013.

Methodology: Spore suspension was prepared by adding 10 ml of 0.1% Tween 80 onto PDA slant of 5 days old culture of Candida rugosa and Geotrichum candidum respectively. Biodegradation of POME was carried out by inoculating 0.1ml (106spores/ml) of respective fungal isolates into different 500 ml Erlenmeyer flasks containing 100ml each of raw POME. They were incubated at 30ºC on a rotary shaker (200rpm). Samples were taken every 24hrs for 144hrs to determine BOD, COD, oil & grease. Similarly, optimization of biodegradation was carried out by studying the effect of different environmental conditions such as different initial pH levels (4.0-8.0), incubation temperature (25-50ºC), concentrations of soy bean (1.5-4.5% w/v) and inoculum size (0.1-0.5 v/v). The experiments were done in triplicates.

Results: Biodegradation studies with selected indigenous fungi showed that C. rugosa was able to remove (44.6%) BOD, (13.9%) COD , (50.7%) oil and grease (O&G) while G. candidum reduced BOD, COD, O&G by 46.9%,16.9% and 64,9% respectively after 144hrs. Optimization of degradation in POME using various environmental and nutrients conditions revealed that at pH 8, C. rugosa showed best degradation of COD (48.6%), BOD (74.5%), O&G (41.8%) removal while COD (59.1%),  BOD (75.7%) , O&G (59.1%) removal  was observed with G. candidum  treatment. The optimal incubation temperature for degradation using each of  fugal isolates was  at 35ºC with 85.2% BOD , 71.8% COD and 67.3% O&G removal for C. rugosa , 87.3% BOD and 63.4% COD for G. candidum  .The best degradation ability for C. rugosa and G. candidum  were demonstrated at 3.5w/v and 2.5w/v soybean concentrations respectively.  The result also showed that increase in inoculum size could not completely reduce oil and grease during degradation process possibly because no single culture supports degradation optimally due to presence of  complex sugars

Conclusion: The selected fungal isolates exhibited high efficiency for removal of oil and grease as well as organic matter from POME but required control of environmental conditions and nutrient expansion for the effective biodegradation of POME.

Open Access Original Research Article

Oropharyngeal Candidiasis in HIV Suspected Patients Attending State Hospital Ijebu-Ode Ogun State Nigeria

O. M. Lakunle, A. Oluwadun, C. O. Adegoke, S. T. Ogunbanwo, H. I. Effedua

Microbiology Research Journal International, Page 1451-1462
DOI: 10.9734/BMRJ/2014/9150

Aims: To determine the effect of CD4 count (a glycoprotein found on the surface of immune cells such as T helper cells, monocytes, macrophages and dendritic cells) on the Candida species associated with Oropharyngeal candidiasis among HIV suspected patients.

Place and Duration of the Study: State Hospital Ijebu Ode Ogun State Nigeria between February 2010 to August 2011.

Methodology: Outpatients attending State Hospital Ijebu Ode were screened for HIV infection using Determine kits, Stat-pak kits and Unigold test kits. Western blot was used to confirm HIV infection and to determine the predominance of HIV specific glycoproteins in HIV seropositive patients. A total of 350 samples of sputum and blood from the HIV seropositive individuals while 300 samples from the HIV seronegative individuals. Sputum samples were cultured on sabouraud dextrose agar, and the isolates were Gram stained while the yeast-like fungi were subjected to germ tube test. CD4 count in blood samples was determined using flow cytometry.

Results: HIV prevalence in females was 70.6% and in males was 29.4%. From three hundred and fifty patients suspected as HIV positive, seventy three had oral candidiasis (20.9%) while 277 (79.1%) were candidiasis negative. Higher oral candidiasis occurred in females (22.7%) than in males (16.5%). Candida alblicans was found to have higher occurrence of 86% among other Candida species. There is no significant association between the occurrence of oral candidiasis and the age of HIV subjects. There was higher occurrence of cases of immune depression (<350 CD4 count) in HIV seropositive (56.3%) than in HIV seronegative (0%) subjects. Candida infections occur when CD4 count was 200-500 cell/µl and usually represent the first indication of immune suppression. Decrease in CD4 count led to increase in occurrence of Candida species. The lowest number of Candida species was recorded when CD4 count was above 300 and Candida alblicans is the most predominant species isolated in this study.

Conclusion: The result of this study shows that HIV infection led to decrease in CD4 count which in turn promotes candidiasis.

Open Access Original Research Article

Impacts of Tetrakis-hydroxymethyl Phosphonium Sulfate (THPS) Based Biocides on the Functional Group Activities of Some Oil Field Microorganisms Associated with Corrosion and Souring

Okoro Chuma Conlette

Microbiology Research Journal International, Page 1463-1475
DOI: 10.9734/BMRJ/2014/11943

Aim: To determine the biocidal efficacy of THPS based biocides currently used in oil fields to control souring and corrosion.

Methodology: By direct monitoring of inhibition of cell growth and inhibition of microbial functional group activities such as the ability to reduce sulfate and generate sulfide by sulfate reducing bacteria (SRB), reduce nitrate to nitrite by heterotrophic nitrate reducing bacteria (hNRB) and oxidation of sulfide and reduction of nitrate by sulfide oxidizing, nitrate reducing bacteria( so-NRB) using CSB-K medium.

Results: We observed that higher doses of THPS (>400 ppm) was required to considerably inhibit the ability of SRB to reduce sulfate and generate hydrogen sulfide. It was also observed that the activities of SRB were more affected by the THPS biocides than those of hNRB and so-NRB.

Conclusion: We conclude that SRB may have developed low level microbial resistance to THPS based biocides as higher doses are required to inhibit their activities. It is therefore recommended that THPS should be used in combination with other biocides or metabolic inhibitors for it to be effective at lower concentrations.

Open Access Original Research Article

Motility of Bacterial Pathogens in Minimal Media Containing Human Serum

N. da S. Mendonça, A. P. Faccio, M. A. Abegg

Microbiology Research Journal International, Page 1476-1490
DOI: 10.9734/BMRJ/2014/11886

Aims: To investigate the effect of human serum on growth pattern, cellular morphology, and motility of potentially pathogenic bacteria. 

Study Design: This was an analytic experimental study.

Place and Duration of Study: Institute of Exact Sciences and Technology (ICET), Federal University of Amazonas (UFAM), Mycology Laboratory, between August 2012 and July 2013.

Methodology: Growth of Escherichia coli, Salmonella typhi, Bacillus subtilis and Bacillus cereus was examined on hard agar (1.5%) solid medium containing 0, 20, or 40 percent pooled human serum. Dilutions of each species were point-inoculated at the center of the plate. Cultivation was carried out aerobically for up to 20 days at 37°C in sealed humidified boxes. Spreading growth was examined by measuring colony diameters, analyzing macro- and micromorphology, and measuring the fractal dimension of colonies.

Results: E. coli and S. typhi strains grew better in Davis and Mingioli agar, whereas B. subtilis and B. cereus grew better in Fujikawa agar. B. cereus and S. typhi developed a white halo of proteolysis around the colony in the medium supplemented with serum. The addition of human serum to minimal hard-agar medium induced a cellular phenotypic change and a colony morphological change, especially in B. cereus and S. typhi. B. cereus and S. typhi developed elongated cells on the colony edge in the presence of human serum, showing cells in raft-like association. Generally, colonies of bacteria grown in the absence of human serum presented smaller fractal and growth dimensions and more-branched spreading, presumably in a sliding translocation.

Conclusion: Cells of “temperate swarmer” species translocated more efficiently on hard agar supplemented with human serum, by sliding and possibly by swarming. The presence of human blood or serum, despite the inhibitory activity of antibodies, may allow pathogenic bacterial cells to overcome the difficulties of low levels of nutrients and hard surfaces with little available water, and may facilitate translocation to other sites. Further investigations of the influence of human serum on swarming and sliding are warranted.

Open Access Original Research Article

Sero-prevalence of HIV, HBV and HCV among Blood Donors in the Kintampo Municipal Hospital, Ghana

Williams Walana, Samuel Ahiaba, Patrick Hokey, Ezekiel Kofi Vicar, Samuel Ekuban Kwabena Acquah, Edmund Muonir Der, Juventus Benogle Ziem

Microbiology Research Journal International, Page 1491-1499
DOI: 10.9734/BMRJ/2014/12160

Aims: This study aimed at establishing the prevalence of some viral Transfusion Transmissible Infectious (TTI) agents among blood donors in the Kintampo North municipality of Ghana.

Study Design: A retrospective cross-sectional hospital based study.

Place and Duration of Study: The study was conducted at the Laboratory unit of the Kintampo Municipal Hospital between May and August, 2013.

Methodology: Archived results (from January 2010 to December 2012) on blood donation from the hospital’s laboratory were reviewed manually. Data comprising age, sex and results on HBsAg, anti-HCV and anti-HIV tests of blood donors were reviewed. The data were analyzed using Microsoft excel 2007 statistical package.

Results: A total of 3402 people were screened for blood donation. Out of this number 3139 (92.3%) were males while 263 (7.7%) were females. The combined sero-prevalence of HBsAg, anti-HCV and anti-HIV was 19.5% (643/3139) and 11.4% (30/263) for males and females respectively. Hepatitis B surface antigen year-on-year prevalence was 9.6%. Anti-HCV and anti-HIV recorded year-on-year prevalences of 4.4% and 4.9% respectively. Donors younger than 20 years recorded the highest prevalence of HBsAg [15.9% (34/214)] followed by those in age group ≥20<30 [10.3% (170/1652)]. The highest prevalence rates of 6.1% and 5.0% for anti-HIV and anti-HCV were observed in age groups ≥50 and ≥30<40 years respectively. The commonest co-infection occurrence was HBV-HCV [45.5% (10/22)].

Conclusion: The prevalence of the viral TTI agents studied among blood donors in the Kintampo municipality is relatively high. Co-infection with HBV and HCV was also high.

Open Access Original Research Article

Antimicrobial Susceptibility Pattern of Some Recently Isolated Serovars of Salmonellae in Jos, Nigeria

Solomon O. Opajobi, Yvonne T. Kandakai-Olukemi, Edmund B. Banwat, Daniel Z. Egah, Solomon C. Chollom, John D. Mawak

Microbiology Research Journal International, Page 1500-1510
DOI: 10.9734/BMRJ/2014/12250

Aim: To determine the salmonellae serovars circulating in North Central Nigeria and their treatability with commonly used antimicrobial agents.

Study Design: Investigative

Place and Duration of Study: Samples were collected and processed at the Jos University Teaching Hospital (JUTH), Plateau State, Nigeria between 2006 and 2011.

Methodology:  Standard  microbiological tests were used for isolation, identification and serotyping of salmonellae from 3509 blood and 5426 stool samples collected from patients attending Jos University Teaching Hospital (JUTH), Jos between 2006 and 2011. Identified serovars were subjected to antimicrobial susceptibility testing using disc diffusion method.

Results: 89 Salmonella isolates were obtained from 8935 samples. Blood and stool cultures yielded 1.4% and 0.70% salmonellae respectively.  The highest number of isolates was from age group 0-9 years 30(33.7%) while patients aged 70 and above accounted for the least number of isolates 1(1.1%).  Males accounted for more isolates 49(55.1%) than females 40(44.9%) (p<0.05). The 89 isolates encountered comprised 39 serovars of which 74(83.1%) were non-typhoidal.  Frequently isolated serovars were         S. typhimurium and S. bargny 11(12.3%) each, S. typhi 7(7.7%) and S. paratyphi B and  S. saint Paul 6(6.7%) each.  Rare serovars isolated included S. Lagos, S. abaSkisii,  S. okerara and S. aminatu  1(1.1) each. All isolates were susceptible to ciprofloxacin and ceftriaxone( MIC≤ 1µg/mL) while more than 50% of the frequently isolated serovars were resistant to chloramphenicol, amoxicillin and cotrimoxazole.

Conclusions: This study reveals a high occurrence of invasive non-typhoidal, multi-drug resistant Salmonella serovars. However, susceptibility to ciprofloxacin and ceftriaxone is completely preserved and can be used for empirical treatment of salmonellosis.

Open Access Original Research Article

Endophytic Fungi: A Poor Candidate for the Production of Lovastatin

V. K. Praveen, S. D. Bhargavi, J. Savitha

Microbiology Research Journal International, Page 1511-1520
DOI: 10.9734/BMRJ/2014/12487

Aim: The aim of the present study was to screen soil and endophytic fungi for production of lovastatin.

Methodology: Soil fungi were isolated by dilution plating technique and endophytic fungi from selected medicinal plants by using standard procedures. All isolates were tested for lovastatin production by Solid State Fermentation (SSF) using wheat bran as substrate.

Results: The soil isolate, Aspergillus terreus NCBI (KM017963) showed positive for lovastatin (1.0 mg/G DWS) whereas none of the endophytic fungi tested showed positivefor lovastatin production.

Open Access Original Research Article

Sensitivity of Salmonella and Shigella to Mangifera indica Linn (Mango) Crude Leaf Extracts

F. J. Okoko, O. O. Akpomie, A. G. Ikejiofor

Microbiology Research Journal International, Page 1521-1530
DOI: 10.9734/BMRJ/2014/11001

Aims: To study the antimicrobial activities of methanol, acetone and aqueous extracts of Mangifera indica Linn crude leaves on Salmonella and Shigella in order to provide natural therapy against them.

Study Design: Salmonella and Shigella isolates were obtained from twenty-five stool samples of hospital patients. Six isolates were identified and characterized. The six isolates were labelled accordingly and used in the study. The antimicrobial activities of the extracts preliminary screened for the presence of phytochemical compounds were tested on the six isolates.

Place and Duration of Study: The study was carried out in the Department of Microbiology, Delta State University, Abraka, Nigeria between October, 2012 and August, 2013. 

Methodology: The test isolates (Salmonella and Shigella) were isolated from stool samples on Salmonella/Shigella agar and characterized. The phytochemical constituents of the leaves of M. indica were extracted by soaking in water, methanol and acetone. The extracts were investigated for antimicrobial activities on theisolates using the agar well diffusion method.

Results: Phytochemical analysis of the leaf extract showed that the extracts contained alkaloids, tannins, flavonoids, steroids, anthroquinones, reducing sugars, glycosides and phenols. The acetone and methanol extracts exerted the highest zone of inhibition (18.00mm and 15.00mm respectively) though there was a significant difference (p<0.05) in the activities of the extracts. There was no significant difference (p>0.05) when compared with that of the water extract. Salmonella was more susceptible to acetone extract than Shigella with a Minimum Inhibitory Concentration of 25.00 mg/ml while that on Shigella was 50.00mg/ml. The MIC of the ethanolic extract on Salmonella was 50.00mg/ml while that of Shigella was 100.00mg/ml.

Conclusion: The acetone and methanolextracts of the leaves of M. indica had high inhibitory effects on Salmonella and Shigella thus providing a basis for its recommendation in the treatment of diseases caused by these organisms.

Open Access Original Research Article

Detection of Pathogenic Legionella pneumophila in Tap Water and Water of Newborn Incubators in Hospitals of Guilan Province by PCR of mip Gene

Masoumeh Ahmadi Jalali Moghadam, Hamidreza Honarmand, Sajad Asfaram Meshginshahr, Bahram Soltani Tehrani, Majid Nojavan

Microbiology Research Journal International, Page 1531-1540
DOI: 10.9734/BMRJ/2014/9830

Aims: This study is designed to determine the frequency of Legionella pneumophila in cold and warm water as well as water containers of newborn incubators in Guilan province hospitals, Iran, using amplification of the macrophage infectivity protein gene (mip gene) by PCR.

Study Design: Cross sectional study.

Place and Duration of Study: The present study was performed in the Cellular and Molecular Research Center, Guilan University of Medical Sciences between June 2011 and July 2012

Methodology:  Samples were collected directly in sterile containers, concentrated in centrifuge, transferred to yeast extract broth containing L- cysteine, Fe2+, Glycin and vancomycin and incubated for 3-4 days. DNA was extracted by using the boiling method and PCR was performed to search Legionella and mip gene using two pairs of primers. Contamination with other bacteria was evaluated in all negative samples using universal primers of 16S rRNA gene.

Results: About 8.5% of the samples had L. pneumophila including 11% of the incubators and 5.8% of both hot and cold tap water.  The mip gene was found in 2.8% of the samples. One third of the incubator and one half of the hot water habited L. pneumophila had the mip gene but it was not found in cold tap water samples. About 87.2% of the negative samples showed bacterial contamination as revealed by PCR with primers of 16S rRNA gene.

Conclusions: This study indicates that in spite of using distilled water for incubators,         L. pneumophila contamination is considerable and other bacterial contamination is very high. It may be related to the length of time that water remains in an incubator container which is a predisposing factor for both biofilm formation and the growth of water microflora. It seems that the high temperature of hot water system and the high rate of free residual chlorine in tap water system are the main causes of low rate of Legionella contamination but are ineffective on contamination with other bacteria.

Open Access Original Research Article

Prevalence and Genotypic Characterization of Extended- Spectrum Beta-Lactamases Produced by Gram Negative Bacilli at a Tertiary Care Hospital in Rural South Western Uganda

Acaku Moses, Freddie Bwanga, Yap Boum, Joel Bazira

Microbiology Research Journal International, Page 1541-1550
DOI: 10.9734/BMRJ/2014/9792

Aim: To determine the prevalence and genotypic characterisation of extended spectrum beta-lactamases produced by gram negative bacilli isolated at Mbarara Regional Referral Hospital (MRRH).

Samples: Gram negative clinical isolates.

Study Design: Laboratory-based descriptive cross-sectional study.

Place and Duration of the Study: MRRH, June and August 2012.

Methods: Gram negative clinical isolates were sub cultured, and identified using biochemical tests. They were screened for ESBL by using oxyimino-cephalosporins and confirmed by double disc synergy Genotyping was performed using the PCR for TEM, SHV and CTX-M. Susceptibility pattern for the extended spectrum beta-lactamases, (ESBL) - positive isolates to other antibiotic classes was performed by the Kirby Bauer Technique.

Results: A total of 484 isolates were included in the study. The commonest ESBL producers were Escherichia coli (34%), followed by unidentified coliforms (19.3%) and Klebsiella spp. (12.7%). Phenotypically, 88/484 were ESBL producers while genotypically 213/ 484 possessed ESBL genes. The ESBL genes were blaCTX-M (146; 70%), blaSHV (72; 34%) and blaTEM (100; 47%). 87of 213 isolates expressed more than one ESBL gene. Of these 36 (7.4%) produced blaCTX-M/blaSHV, 28 (5.8%) blaCTX-M /blaTEM, 4 (0.8%) blaSHV/ blaTEM and 19 (3.9%) blaCTX-M/blaSHV/blaTEM. Sixty two (16%) were phenotypically and genotypically positive, 12 (3%) of the isolates were phenotypically positive but genotypically negative and 140 (37%) isolates were phenotypically negative but genotypically positive. The ESBL producers were highly susceptible to imipenem (95%), nitrofurantoin (66%) but less susceptible to ampicillin (4%) and ticarcillin (7%).

Conclusion: ESBL production among the Gram-negative clinical isolates at MRRH is very high with several isolates possessing multiple genes. The ESBL producers are highly susceptible to imipenem, but very resistant to ciprofloxacin.

Open Access Original Research Article

Study on Association of Recent and Past Chlamydophila pneumoniae Infection with Classic Multiple Sclerosis

Hamidreza Honarmand, Masoumeh Ahmadi Jalali Moghadam, Hamidreza Hatamian, Ali Roudbary

Microbiology Research Journal International, Page 1575-1584
DOI: 10.9734/BMRJ/2014/9831

Aims: We conducted this study to determine if there is any correlation between Classical Multiple Sclerosis and Chlamydophilia pneumoniae infection by ELISA (IgM, IgG, IgA).

Study Design: cross sectional study

Place and Duration of Study: The present study was performed in the Department of Microbiology, Guilan University of Medical Sciences   between April 2012 and April 2013

Methodology: Chlamydophila pneumoniae infection certified by ELISA in patients (n=46) and control (n=46) using commercial assays (anti- C. pneumoniae IgG, anti- C. pneumoniae IgM, and anti- C. pneumoniae IgA kits). Data were analyzed by using four statistical tests (Pearson chi square, Kendall's tau, and Spearman's rho).

Results: Seropositivity to anti- C. pneumoniae IgG was seen less frequently in patients versus controls (69.0% versus 81.4%; P=0.187). Seropositivity to anti- C. pneumoniae IgA  was also observed less frequently in patients than in controls (7.2 % versus 11.6%; P= 0.479).However anti- C. pneumoniae IgM antibodies were seen more often in classical multiple sclerosis patients than it was in controls ( 11.9% versus 2.3%; P= 0.085).

Conclusion: We concluded that recent or past C. pneumoniae infection has no correlation in initiation or protection of CMS.

Open Access Original Research Article

Potential of Probiotic Lactobacillus plantarum 2621 for the Management of Infertility

Praveen Bhandari, Praveen Rishi, Vijay Prabha

Microbiology Research Journal International, Page 1585-1596
DOI: 10.9734/BMRJ/2014/12129

Background: Infertility outcomes may be associated with the infections that would lead to morphological defects of spermatozoa in vitro. The purpose of this work was to investigate the effect of Lactobacillus plantarum 2621 (L. plantarum) on adherence of sperm agglutinating Escherichia coli (E. coli) in vitro and in vivo as well as its effect on fertility outcome.

Materials and Methods: Interference of E. coli adherence to vaginal epithelial cells (VECs) by L. plantarum was studied by carrying out different assays such as exclusion, competition and displacement. Further, in vivo study was carried out in mouse model to evaluate the effect of presence of L. plantarum against E. coli and its effect on fertility outcome by administering intravaginally at one hour interval between L. plantarum (108c.f.u./20µl) and different concentrations of E. coli (102,104, and 106 c.f.u/20 µl) for ten consecutive days.

Results: 116•8 bacteria/VEC adhesion levels were observed for L. plantarum 2621 whereas values for E. coli were 60•5 bacteria/VEC. L. plantarum interfered to different extents with the adherence of E. coli. L. plantarum 2621 decreased the adhesion by displacement and competition in a significant level (90.3% and 68.5% of inhibition). L. plantarum 2621 also excluded the E. coli attached to VEC (25.8% of inhibition).Upon mating and completion of gestation period 100% fertility was observed with 108c.f.u./20µl L. plantarum and 102 c.f.u/20µl E. coli, whereas 100% females were infertile when administered with 106 c.f.u/20µl of E. coli alongwith 108c.f.u./20µl L. plantarum and only 50% fertility outcome was observed with 104 c.f.u/20µl E. coli.

Conclusion: Results indicated that L. plantarum displaces colonization of E. coli and endows competition that resulted in reinforcement of natural microflora and affects fertility outcome depending on the presence and count of E. coli.

Open Access Review Article

Brucellosis: A Neglected Zoonosis

B. Y. Kaltungo, S. N. A. Saidu, I. W. Musa, A. Y. Baba

Microbiology Research Journal International, Page 1551-1574
DOI: 10.9734/BMRJ/2014/11061

Brucellosis is a bacterial disease mainly of domestic animals. The infection is directly transmitted to humans by animals through breaks in the skin or by contact with infected materials like aborted foetuses and placenta. It can also be transmitted indirectly by ingestion of contaminated animal products as well as inhalation of the agent. It is an important zoonosis worldwide which accounts for about 500, 000 reported human cases annually around the globe; particularly amongst agricultural and pastoral populations. It results in serious economic losses in animals due to abortion, reduced fertility, birth of weak off springs and reduced productivity. In humans, it leads to chronic debilitation resulting in low work output and subsequent negative economic impact. The paper reviews brucellosis in different species of animals. It highlights the aetiology, morphology, host range, pathophysiology, clinical signs, pathology and epidemiology of the disease in various species. Preventive and control measures against the disease, economic and public health implications have also been examined. It is concluded that, the eradication of brucellosis in animals may be achieved by long-term investment in surveillance programmes, including testing and culling of positive reactors. Vaccination of animal hosts may culminate in the eradication of the disease in human population.