Open Access Original Research Article

Molecular Characterisation and Antagonistic Activity of Fructophilic Lactic Acid Bacteria Isolated from Selected Fruits

Olateru Comfort Tosin, Ogunbanwo Samuel Temitope

Microbiology Research Journal International, Page 1-17
DOI: 10.9734/MRJI/2018/44484

Fructophilic lactic acid bacteria have very unique biochemical characteristics when compared to other LAB in relation to their hexose sugar metabolism and utilization of oxygen. The aim of this study was to isolate and identify lactic acid bacteria from fresh fruits and determine their antibacterial activity against pathogenic and spoilage organisms.84 fresh fruit samples comprising of Mango, Apple, Oranges, Watermelon, Guava, Lime were purchased from major fruit markets. Lactic acid bacteria were isolated from fruits using modified Man Rogosa Sharpe media and phenotypically identified using Gram staining, biochemical tests and API 50CH Kit. Polymerase Chain Reaction (PCR) analysis and 16SrRNA sequences were used to confirm their identity. Antibacterial activity of the cell free supernatant of the LAB was carried out against four pathogenic and two spoilage bacteria respectively. The result obtained indicate that one hundred and ninety fructophilic lactic acid bacteria were isolated and identified into four groups: Pediococci, Lactobacilli, Enterococci and Leuconostoc respectively. Lactobacillus plantarum had the highest frequency of occurrence while Lactobacillus alimentarium had the least. Out of 190 FLAB isolates obtained, only 98 showed inhibitory activity against the pathogenic and spoilage organisms tested, only 23 of the tested isolates were able to produce bacteriocin as shown by the effect of enzyme treatments. This study clearly suggests that Fructophilic LAB share similar traditional attributes with those obtained from fermented and dairy food, hence the need to utilize them in food processing and preservation.

Open Access Original Research Article

Highly Resistant Staphylococcus aureus Isolated from Patients Attending a Tertiary Hospital, South Western Uganda

Andrew Baguma, Julius Tibyange, Tonny Jimmy Owalla, Atek Atwiine Kagirita, JudeBusingye ., Joel Bazira

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/MRJI/2018/44939

Background: S. aureus is a frequent cause of human infections and is one of the most important nosocomial pathogens. Widespread antimicrobial resistance has limited therapeutic choices to treat S. aureus infections. Methicillin-resistant S. aureus has continued to cause significant infections today challenging public health initiatives to a better healthcare.

Materials and methods: This was a cross-sectional study carried out between February 2016 and January 2017 among inpatients at Kabale Regional Referral Hospital, Various clinical specimens were collected basing on nature of infection, and analysed using standard phenotypic methods to characterise S. aureus. Presence of MecA gene was detected by PCR to confirm methicillin resistance strains. Antibiotic susceptibility tests were done to determine resistance patterns using standard methods.

Results: The study revealed the prevalence of S. aureus at 86.7% (n = 390) among inpatients, where; 223(57.2%) were Methicillin-resistant S. aureus. Methicillin-resistant S. aureus was predominantly isolated from surgical sites; 56.9 %( n = 166), Road traffic accident wounds 67.9% (n = 19) and Burn wounds; 56.3% (n = 09).

Conclusion: There was high prevalence of S. aureus among inpatients and majority of the isolates were methicillin-resistant. Methicillin resistant strains were highly resistant to multiple antibiotics that are commonly used.

Open Access Original Research Article

Molecular Detection of Astrovirus in Diarrhoeic Stools of Children in North East Nigeria

Oyinloye, S. Oyebode, Aminu, Maryam, Elijah E. Ella, Nimzing, Lohya

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/MRJI/2018/42815

Background: Human astroviruses are a leading cause of severe viral gastroenteritis and are responsible for at least 95% of nonbacterial gastroenteritis outbreaks throughout the world.

Methods: Six hundred (600) diarrhoeic stools of children under 5 years were collected between May 2013 – April 2014 and screened for astrovirus using a 3rd generation Ridascreen ELISA kit (R-Biopharm AG, Germany). Demographic data were collected via a questionnaire. Analysis of the data was done using online Easy-Chi-square (p<0.05) statistical package. The ELISA positive astrovirus samples were further analysed by reverse transcription polymerase chain reaction (RT-PCR) and amplicons generated were sequenced. Phylogenetic tree of sequences was constructed using the Neighbour-Joining Model with 1000 replicate bootstrap value in MEGA 6.0.

Results: Astrovirus prevalence of 5.0% (30/600) was obtained. The prevalence of astrovirus in Taraba, Bauchi and Borno states was 5.5% (11/200), 4.5% (9/200) and 5.0% (10/200), respectively. Of the 30 astrovirus positive samples, 63.3% (19/30) were male and 36.7% (11/30) female. Female children were more likely to be infected with astrovirus (OR= 1.38; 95% CI) compared to male children. The highest astrovirus prevalence (8%: 9/112) and lowest (1.9%: 1/54) prevalence were in children 1-2 years and 0-6 months respectively. Most children were infected before 2 years. Of the 30 astrovirus, ELISA positive samples analysed by RT-PCR, 5 (16.5%) amplicons of ORF genes with 400bp were seen and subsequently sequenced.

Conclusion: Sequence analysis showed that all the strains were HAstV-5 indicating the strain prevalent in the study area. The results of the present study suggest that astrovirus contribute significantly to the disease burden of childhood diarrhoea in parts of North Eastern Nigeria.

Open Access Original Research Article

Fungi Isolated from Poultry Droppings Express Antagonism against Clinical Bacteria Isolates

Stephen Nnaemeka Ezekwueche, Chinelo Ursula Umedum, Chibuzo Christain Uba, Ifeoma Sandra Anagor

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/MRJI/2018/46183

Aim: This study was conducted to isolate antibiotic producing fungi from poultry droppings.

Place and Duration of Study: Poultry droppings were collected from poultry farms and markets within Ihiala Local Government Area, Anambra State between March 2017 and October 2017.

Methodology: One hundred and fifty poultry dropping samples were analyzed. One (1) g of fresh poultry droppings were weighed and serially diluted. The dilutions were cultured on Sabouraud Dextrose Agar and Potato Dextrose Agar for five days. Isolates were characterized morphologically and microscopically. Isolates showing antagonism were subjected to submerged fermentation. The Screening and determination of minimum inhibitory concentration (MIC) of the secondary metabolite extracts was done using agar well diffusion method. The isolates were screened for antagonism against four bacteria isolates namely, Escherichia coli, Salmonella typhi, Bacillus subtilis and Staphylococcus aureus.

Results: Five isolates namely, Aspergillus niger, Aspergillus tubingensis, Rhizomucor variabilis, Aspergillus aculeatus and Candida rugosa were identified. Aspergillus tubingensis and Rhizomucor variabilis showed antagonism against the test bacteria during preliminary screening. Aspergillus tubingensis and Rhizomucor variabilis showed antagonism against Bacillus subtilis. After fermentation, the secondary metabolite extracts from Aspergillus tubingensis and Rhizomucor variabilis, were active against Bacillus subtilis at different concentrations with MIC of 20.27mg/ml and 12.72mg/ml respectively.

Conclusion: The extracts from two fungal isolates namely; Aspergillus tubingensis and Rhizomucor variabilis exhibited antagonism against Bacillus subtilisonly. The extracts when purified, may serve as a new drug molecule produced from natural source.

Open Access Original Research Article

Phenotypic Study on Some Virulence Factors and Molecular Screening of Aminoglycoside Resistance among Klebsiella pneumoniae Strains Isolated from Urinary Tract Infections in Pediatric Cases in Egypt

Rasha Abd El-Hamid Alm El-Din, Mohamed Galal El-Sanosy

Microbiology Research Journal International, Page 1-11
DOI: 10.9734/MRJI/2018/46061

Aim of the Work: The aim of this study was to identify aminoglycoside resistance among Klebsiella pneumoniae strains isolated from cases of urinary tract infections in pediatrics and to evaluate the presence of 16S rRNA methylase genes including armA and rmtB genes in the isolated strains.

Materials and Methods: Ninety-eight Klebsiella pneumonia strains were isolated and identified by conventional microbiological methods, 65.3% isolated from cases with cystitis and 34.7% from cases with pyelonephritis. Antimicrobial susceptibility testing was done using disc diffusion test. Presence of 16S rRNA methylase genes including armA and rmtB genes were detected by polymerase chain reaction [PCR].

Results: The overall aminoglycoside resistance was 48%. Individually, the rate of resistance against Amikacin and Gentamycin were 22%, 26% respectively. Also, 73.5% were multidrug-resistant [MDR], 20.4% were extensive drug resistant [XDR] and 61.2% strains were extended-spectrum β-lactamases [ESBL] producers. PCR amplification of 16S rRNA methylase genes revealed that armA was the most prevalent gene detected in 30 isolates; while rmtB was detected in only 17 cases.

Conclusion: This finding highlights the alarmingly increase in multidrug-resistant Klebsiella pneumoniae isolates with high frequency of 16S rRNA methylase genes. Consequently, following a wise antibiotic policy is critically necessary to limit the potential spread of resistant genes.