Open Access Original Research Article

Evaluation of in vitro Anti-proliferative Activity of L-arginine deiminase from Novel Marine Bacterial Isolate

Rahamat Unissa, M. Sudhakar, A. Sunil Kumar Reddy

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/BMRJ/2016/23592

L-Arginine deiminase is a therapeutic l-arginine depleter found to counteract various arginine auxotrophic cancer cells (do not express ASS/OCT). The aim of the present study was to evaluate the anti -proliferative activity of the purified l-arginine deiminase from Vibrio alginolyticus 1374.  Production of the enzyme was carried out by shake flask method under optimal conditions. The enzyme thus produced was purified to near homogeneity by ammonium sulphate fractionation followed by ion exchange and gel filtration chromatography. The enzyme was purified to 529.43 fold and showed final specific activity of 280.6 IU/mg with 43.5% yield. SDS-PAGE revealed that the purified enzyme had a molecular weight of 48 kDa. The purified enzyme tested against A375-C6, MCF-7, HCT-113 and Jurkat, clone E6-1cell lines showed I.C.50 value of about 5. 21, 6.3,8 and 3.13 U/ml respectively. ADI obtained from Vibrio alginolyticus 1374 has several beneficial properties needed for its use as a therapeutic and industrial enzyme. It was active over wide range of pH (4-10) and temperature (25°-50°C), had high substrate specificity etc. Moreover it showed broad range of activity against cancer cells lines of human origin. Further animal model studies must be carried out to develop ADI as a potential drug.

Open Access Original Research Article

Hand Contamination among Food Handlers

H. K. Allam, M. A. Al-Batanony, A. S. Seif, E. T. Awad

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/BMRJ/2016/24845

Introduction: Food handlers act as a vehicle for microorganisms causing a potential risk to the public health. Hands contamination of food handlers can be used as an indicator of their behavior regarding food-related practice and personal hygiene.

Aim of the Study: To assess hand contamination among food handlers working in Menoufia University and Shebin Al-Kom Educational hospital kitchens and the effect of a health education session over them.

Subjects and Methods: A seventy two food handlers were the target group of this study. A pre-structured questionnaire was filled including socio-demographic data beside questions regarding risk factors for contamination and personal hygiene practices during dealing with food. Hand rinse sample was taken from each participant to detect the contaminants. A brief and simple health education session was held regarding personal hygiene practices. Three months later, the same kitchen was revisited to appraise the participants’ hygiene compliance where another hand sample rinses were taken.

Results: Both Staphylococcus Epidermidis and Escherichia coli (41.7%) followed by Staphylococcus aureus (29.2%) were the prevalent organisms. A significant reduction in hand contamination was achieved after the interventional health education session. Being illiterate, ignoring hand wash after toilet or touching dirty materials and having long fingernails were the significant risk factors for hand contamination.

Recommendations: periodic interventions on food safety, proper hand washing procedures and good hygiene practices beside regular medical checkup are necessary to improve food handlers’ hygiene practices.

Open Access Original Research Article

Detection and Molecular Identification of Persistent Water Vessel Colonizing Bacteria in a Table Water Factory in Nigeria

O. Nwaiwu, M. I. Nwachukwu

Microbiology Research Journal International, Page 1-12
DOI: 10.9734/BMRJ/2016/24378

Aims: To establish organisms persistent in a table water production facility and to determine the points they entered in order to eliminate any risk of having pathogens in a water production system.

Study Design: Analytic observational studies

Place and Duration of Study: Bottling facility in Nigeria and University of Nottingham, United Kingdom. Study was between August 2011 and Sept 2013.

Methodology: Sample control points were subjected to counts of bacteria, yeasts, and mold using membrane filtration to trace the source of increased bacteria counts in a table water production factory. Organisms were identified by 16S rRNA sequencing and biofilm formation was assessed with micro titer dish biofilm formation analysis.

Results: Total bacteria and Pseudomonas Spp. count were highest in the carbon filter and 5% (v/v) chlorine used for disinfection was found to be effective against planktonic cells of 18 hour cultures of Pseudomonas Spp. isolated from the carbon filter tank. No yeasts and mold were detected and after a sand blasting exercise to clean affected tanks, total bacteria counts on tryptone glucose extract medium decreased to less than factory allowable limits of 25 cfu per 100 ml of water with no further growth of Pseudomonas Spp. on centrimide medium. Other bacteria that emerged after the sand blasting exercise were regarded as persistent but had less biofilm forming ability (p=.02) when compared with Pseudomonas aeruginosa PAO1. The persistent bacteria identified after sequencing were Aeromonas hydrophila and Serratia proteamaculans.

Conclusion: Apart from Pseudomonas Spp., other bacteria can persist in the water tanks of a water bottling facility and routine checks may fail to detect an underlying problem until it becomes obvious. Prompt corrective action ensures that public safety is not compromised. This was the first time Aeromonas hydrophila and Serratia proteamaculans were identified from the water tanks in the bottling facility.

Open Access Original Research Article

Endophytic Microorganisms in Leaves of Moringa oleifera Collected in Three Localities at Pernambuco State, Northeastern Brazil

Igor F. A. C. Souza, Thiago H. Napoleão, Kêsia X. R. F. de Sena, Patrícia M. G. Paiva, Janete M. de Araújo, Luana C. B. B. Coelho

Microbiology Research Journal International, Page 1-7
DOI: 10.9734/BMRJ/2016/24722

Aims: This work investigated the population density of endophytic microorganisms from Moringa oleifera leaves collected in three localities at the state of Pernambuco (northeastern Brazil): Urban (campus from the Federal University of Pernambuco, UFPE) and forest (botanical garden) areas at Recife city and an urban area (industrial district) at Caruaru city.

Place and Duration of Study: Department of Antibiotics and Department of Biochemistry from Federal University of Pernambuco, between July 2014 and July 2015.

Methodology: Sodium hypochlorite was used to disinfect the leaves, which were macerated in PBS buffer and separately sowed on seven culture media containing antibacterial or antifungal agents.

Results: The most of endophytes isolated were bacteria and the highest density was found in leaves from the forest area. With respect to the fungi, there was no statistical difference between the density in leaves from UFPE campus and the botanical garden while no fungal isolates was obtained from leaves collected in Caruaru. The highest diversity of endophytes was found in the leaves from the botanical garden, with 111 different isolates. A total of 71, 94 and 50 bacterial isolates were obtained from leaves of UFPE campus, botanical garden and Caruaru, respectively. The number of fungal isolates were 17 (UFPE campus) and 12 (botanical garden).

Conclusion: In conclusion, the methodology employed in this work was effective for the isolation of endophytes; climatic and geographical conditions may interfere in density and diversity of endophytes from M. oleifera leaves.

Open Access Original Research Article

Phenotypic Characterization of Toxigenic Fungi, Evaluation and Effect of Post Treatment on Aflatoxin and Ochratoxin A Content in Smoked Fish and Smoked Bush Meat

Fowoyo Patience Temitope, Bodunde Rachael Oluwaseun

Microbiology Research Journal International, Page 1-8
DOI: 10.9734/BMRJ/2016/24036

Smoked fish and smoked bush meat constitute a major food for humans and are sometimes consumed without further processing leading to contamination with toxins such as aflatoxin and ochratoxin A in which consumption may have serious impact on public health. This study is aimed at detecting the incidence of aflatoxin and ochratoxin A in smoked fish and smoked bush meat samples in Lokoja, Kogi state. A total of thirty-six (36) samples of smoked fish and smoked bush meat were examined; Eight (8) of which belonged to different species of smoked fish (catfish and tilapia fish) and four (4) different species of smoked bush meat samples purchased in triplicates from Old market, Lokoja. All the fish samples contained ochratoxin A with a content of between 2.3-11 µg/kg while the smoked bush meat samples had ochratoxin A content of 3.8-11.5 µg/kg. Aflatoxin content in the fish samples ranged between 4.8-12.5 µg/kg while in smoked bush meat it had 4.5-13.8 µg/kg. Aspergillus flavus, Penicillium sp., Mucor sp. and Aspergillus niger were the fungi isolated from the samples. Aspergillus niger, Penicillium sp. and Aspergillus flavus produced ochratoxin A while only Aspergillus flavus produced aflatoxin. The smoked fish and smoked bush meat samples were subjected to heat and brine treatments to determine their ability to reduce the concentration of the toxins. After heat treatment of the samples, aflatoxin concentration ranged between 0.019-0.067 µg/kg and for ochratoxin A, the range was between 0.011-0.06 µg/kg. Aflatoxin concentration ranged between 0.017-0.065 µg/kg and ochratoxin A concentration ranged between 0.057 - 0.108 µg/kg after brine treatment and this indicated that both treatments drastically reduced the aflatoxin and ochratoxin A content of the samples though heat treatment was more effective. It is therefore recommended that consumption of smoked fish and smoked bush meat as ready-to-eat without undergoing post treatment be discouraged so as to decrease the likelihood of the occurrence of food intoxication. Also the use of high temperatures of about 80°C or soaking in brine solution (7.5%) should be encouraged for post treatment of these foods before consumption.

Open Access Original Research Article

Species Distribution and Drug Susceptibility of Candida Isolates from Various Clinical Specimens at a Tertiary Care Hospital in Kashmir

Asifa Nazir, Farhath Kanth, Anjum Farhana

Microbiology Research Journal International, Page 1-6
DOI: 10.9734/BMRJ/2016/24548

Aims: The aim of our study was to identify the distribution of Candida species among clinical isolates and their sensitivity pattern for common antifungal drugs.

Study Design: Prospective observational study.

Place and Duration of Study: Department of Microbiology, Government Medical College Srinagar, Kashmir, India, from December 2014 to January 2016.

Methodology: Identification of hundred and five different Candida species as well as antifungal sensitivity testing was performed with Vitek2 Compact (Biomerieux France) using vitek 2 cards for identification of yeast and yeast like organisms (ID-YST cards).

Results: Among the 105 culture positive isolates, 35 (33.3%) were C. albicans and 70 (66.6%) were Non Candida albicans (NCA). Among NCA, 35 (50%) were C. tropicalis followed by other species. All the Candida isolates were sensitive to micafungin and capsofungin whereas the susceptibility pattern of amphotericin B varied from 75.6% to 100% and the highest rates of resistance were seen for fluconazole.

Conclusions: Infections caused by Candida sp are on the rise. Hence accurate identification and susceptibility pattern is necessary for management of all Candida infections.