Open Access Short Research Article

Tolerance Tests of Alcaligenes faecalis BW1 Extract

Ilham Zahir, Abdellah Houari, Mohammed Iraqui, Saad Ibnsouda

Microbiology Research Journal International, Page 905-917
DOI: 10.9734/BMRJ/2014/9715

Aims: To highlight whether metabolites of Alcaligenes faecalis BW1 extract can be administered orally for their possible antimycobacterial effects.

Study Design: Study of the influence of certain parameters on the extract of Alcaligenes faecalis by using either discs or well diffusion methods against M. smegmatis.

Place and duration of study: Laboratory of Microbial Biotechnology, Department of Biology, Faculty of Sciences and Technical, University Sidi Mohamed Ben Abdellah, BP 2202, Road of Immouzer, Fez, Morocco. From April to August, 2012.

Methodology: The impact of acidic pH of gastric juice, bile, hydrogen peroxide, pancreatic enzymes and lysozyme on the antimycobacterial activity of Alcaligenes faecalis BW1 extract was evaluated by agar diffusion method. Detection whether or not antibacterial metabolites having a synergistic effect with rifampicin against M. smegmatis was also explored.

Results: Antibacterial metabolites of Alcaligenes faecalis BW1 extract resist to the action of gastric pH, gallbladder bile and hydrogen peroxide. In addition, they are not affected by pancreatic enzymes and lysozyme. Moreover, they have a synergistic effect with rifampicin against M. smegmatis.

Conclusion: Anti-mycobacterial metabolites of Alcaligenes faecalis BW1 extract are compatible with rifampicin and could be administered orally as antitubercular agents after their purification, identification in further work

Open Access Short communication

Antibacterial Activity of Two Brown Algae (Cystoseira compressa and Padina pavonica) Against Methicillin-Resistant Staphylococcus aureus

Gorkem Dulger, Basaran Dulger

Microbiology Research Journal International, Page 918-923
DOI: 10.9734/BMRJ/2014/10449

Aqueous and ethanolic extracts obtained from two brown algae (Cystoseira compressa (Esper) Gerloff et. Nizamuddin) and Padina pavonica (Linnaeus) Thivy) have been investigated for their ability to inhibit 35 hospital isolates of methicillin-resistant Staphylococcus aureus (MRSA). Both aqueous and ethanolic extracts of the plant were effective on MRSA. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) values of the ethanolic extract with the greatest antibacterial activity were those of Cystoseira compressa MIC 3.2-6.3mg/mL and MBC 6.3-25mg/mL, respectively

Open Access Original Research Article

Molecular Characterization of Hepatitis B Virus Circulating in Asymptomatic and Symptomatic Carriers in Côte d’Ivoire from 2010 to 2013

Doumbia Moussa, A. Kouassi M. Bengue, M. Dosso, D. Sevede, S. Kakou N. Gazoa

Microbiology Research Journal International, Page 831-840
DOI: 10.9734/BMRJ/2014/7851

Background: Africa and Asia remain the continent most affected by viral hepatitis B with more than1 million deaths per year. These deaths are due to complications such as cirrhosis and hepatocellular carcinoma. Several studies have shown that the rate of progression of hepatitis B to cirrhosis and liver cancer is related to the virus genotypes. Previous analyses of hepatitis B virus genome have revealed 10 genotypes (A-J) with distinct geographical distribution worldwide. Some studies have shown that the genotype E is predominant in West Africa. In Côte d'Ivoire, few data exist on the genotypes circulating. The presence of genotypes A, B, C and E has been proven but not their involvement in the development of liver complications.

Aim of Study: To determine the hepatitis B virus genotypes circulating in asymptomatic and symptomatic carriers and to establish correlation between genotypes and clinical outcome in Côte d’Ivoire.

Place and Duration of Study: Patients were recruited in different hospitals in Côte d’Ivoire and study was conducted in the National Reference Center for Viral Hepatitis of the Institute Pasteur from April 2010 to February 2013.

Methodology: The study examined samples from 754 subjects using serological and molecular techniques. PCR and multiplex-nested PCR, using type-specific primers, were carried out to determine genotypes of hepatitis B virus in the study samples. 

Results: Hundred thirty nine were HBsAg-positive. Out of the 139, 49% were asymptomatic and 51% were symptomatic. Among the HBsAg-positive, the average age was 41years with 38.85% having HBV DNA in their blood samples. Sixty-four point eight percent of the latter were typeable with 97.1% as genotype E and 2.9% as genotype B. Conclusion: This study revealed a predominance of genotype E of HBV and revealed that genotype E was associated (P=0.03) with clinical Outcome.

Open Access Original Research Article

Antimicrobial Activity of Bacteria Associated with Seaweeds against Plant Pathogens on Par with Bacteria Found in Seawater and Sediments

T. Suvega, K. Arunkumar

Microbiology Research Journal International, Page 841-855
DOI: 10.9734/BMRJ/2014/8678

Aim: We report antimicrobial activities of bacteria associated with 10 native and one invasive species of seaweeds on par with bacteria found in the seawater and sediment. Bacteria exhibiting antimicrobial activity were phylogenetically analysed using 16S rRNA gene.

Place and Duration of Study: Samples of seaweeds, seawater and sediments collected at 6 localities of south east coast of India between December 2009 and January 2010 during monsoon season.

Methodology: Culturable bacteria in seaweeds (epibiotics and endobiotics), seawater and sediments were isolated through serial dilutions using 1.5% ZoBell marine agar (HiMedia, India). Bacterial isolates producing antibiotics were identified by screening against commercial antibiotics and they were subjected to morphological, Gram’s staining and biochemical studies. Chemical property and stability of antimicrobial substances obtained from the promising bacteria active against plant pathogens were studied. Phylogenetic analysis of antibiotics-producing marine bacteria was made using 16S rRNA gene sequencing technique.

Results: A number of673 isolates obtained through the isolation process were found to be the member of 27 bacterial genera, with species of Bacillus recording a maximum of 40.2%. Generally species of bacterial isolates in the association (seaweeds: epibiotics, 39.54% and endobiotics, 40.74%, seawater 8.61% and sediments 11.11%) produced antibiotics and active against plant pathogens (Xanthomonas axonopodis pv. citri, X. oryzaepv. oryzae and Ustilaginoidea virens) were associated with seaweeds (epibiotics 33.46% and endobiotics 43.11%) and sediments (23.43%). Extracellular components of active bacteria are proteins and retaining bioactivity at pH 7.0, up to 40°C and antifungal property up to 60°C. Extracts obtained from the active bacteria are nonpolar lipophilic substances exhibited only antifungal activity.

Conclusion: Bacterial population were considerably higher in seaweeds as compared to seawater and sediments, and at the same time higher bacterial population was recorded in Gulf samples than the open coast samples. Most of the bacterial isolates  associated with seaweeds were found to produce antibiotics.

Open Access Original Research Article

The Effect of Ionizing Radiation on Multi-drug Resistant Pseudomonas aeruginosa Isolated from Aquatic Environments in Egypt

S. M. Ezzat, M. A. Abo-State, H. M. Mahdy, E. H. Abd El- Shakour, M. A. El-Bahnasawy

Microbiology Research Journal International, Page 856-868
DOI: 10.9734/BMRJ/2014/7606

Aims: This study was conducted to determine the effect of different doses of gamma radiation on Multi-drug resistant Pseudomonas aeruginosa isolated from River Nile at Rosetta branch and associated drains in Egypt.

Place and Duration of Study: The study was started with samples collection in August 2010 through April 2011 in the Microbiology Dep., Central Laboratory for Environmental Quality Monitoring (CLEQM), National Water Research Center (NWRC), Cairo, Egypt and the National Centre for Radiation Research and Technology (NCRRT), Egyptian Atomic Energy Authority (EAEA), Cairo, Egypt.

Methodology: Water samples were processed using membrane filtration, 144 strains of P. aeruginosa were isolated and identified and their antibiotic susceptibility was determined against 20 different antibiotics using agar disc diffusion method. Irradiation of bacterial isolates was processed using gamma irradiation unit of cobalt (Co60) and the D10-value was calculated from the survival curve.

Results: Isolates were categorized as multi-drug resistant Pseudomonas aeruginosa (MDRPA). 125 (86.8%) were found to be extensively drug resistant (XDR) and 19 (13.2 %) were characterized as possible pan drug resistant (PDR). The highest resistance (100%) was mostly directed to amoxycillin/clavulanic acid, ampicillin, carbenicillin, methicillin, cephalothin, kanamycin, vancomycin, tetracycline, erythromycin, clindamycin, trimethoprim/sulfamethoxazole, nitrofurantoin and chloramphenicol. More than 75% of isolates were sensitive to norfloxacin (82.6%), piperacillin (81.2%), amikacin (79.2%) and tobramycin (77.8%). 63.2%, 26.4% and 14.6% of isolates were sensitive to ofloxacin, cefotaxime and ceftriaxone, respectively. The viable counts of MDRPA decreased with increasing radiation doses of gamma rays up to the lethal dose (3 kGy). The counts of 0.5, 1.0, 1.5, 2.0, 2.5 kGy irradiated samples were respectively 7.8, 6.5, 4.7, 2.3& 1 log10 and the D10-value calculated from the survival curve was 0.27 kGy.

Conclusion: Contaminated fresh water may act as reservoirs for antibiotic resistant pathogens. Regular monitoring of Multi-drug resistant pathogens in aquatic environments should be adopted constantly. Gamma radiation demonstrates a potential value for wastewater treatment and pollution control.

Open Access Original Research Article

Phytochemical and Antimicrobial Analysis of Callus Extracts of Biophytum sensitivum (Linn) DC

Sirigiri Chandra Kala, Muvva Vijayalakshmi, Shaik Ibrahim Khalivulla, Kokkanti Mallikarjuna

Microbiology Research Journal International, Page 869-884
DOI: 10.9734/BMRJ/2014/7270

Aims: In vitro studies are highly instrumental in selecting a drug for a particular disease and also in getting the preliminary evidence to proceed for further In vivo pharmacological research. Hence, the study is designed to screen and identify the therapeutic suitability of this plant extract for the treatment of a particular disease. And to find out the presence of phytochemicals and antimicrobial activity of leaf callus cultures of Biophytum sensitivum Linn.

Place and Duration of Study: Department of Botany and Microbiology, Acharya Nagarjuna University, Nagarjuna Nagar, Guntur 522510, India during June 2010 to Dec 2010.

Methodology: Here we induced the callus from the leaf explants of this species on Murashige and Skoog basal medium supplemented with various concentrations of BA and NAA. BA 1.0 mg/l with NAA 1.0mg/l is the best concentration for optimal results. The callus was extracted sequentially with hexane, chloroform, ethyl acetate and methanol for 24h by using Soxhlet apparatus. These extracts were used to investigate the presence of phytochemicals which was performed according to the Aiyelaagbe and Osamudiamen [29] and Egwaikhide et al. [30] methods. The mean values were statistically analyzed with the MINITAB 14 by the general one way (un stacked) analysis of variance (ANOVA) to find out the most effective extracts

Results: The qualitative phytochemical analysis of various solvent extracts showed the presence of phytochemicals viz., Terpenoids, phenols, flavonoids, saponins, quinones and phenols. All the extracts except hexane  showed highest zone of inhibition against gram positive and gram negative bacteria (4.46-22.9mm) as well as fungi (7.64-144.4mm) by agar well diffusion method at 100ppm concentrations. The results of present study indicate that the callus of this plant is a potential source of antimicrobial agents and drugs and need to be investigated further.

Conclusion: From the present study, it is evident that, the antibacterial active constituent of Biophytum sensitivum is having a constant expression pattern over different pathogens. This plant leaf callus can be further subjected to enhancement and isolation of the therapeutic antimicrobials and carry out further pharmacological evaluation

Open Access Original Research Article

Improved Cultural Conditions for Methionine Accumulation in Submerged Cultivation of Bacillus cereus S8

V. N. Anakwenze, C. C. Ezemba, I. A. Ekwealor

Microbiology Research Journal International, Page 885-895
DOI: 10.9734/BMRJ/2014/8224

Aims: To improve the cultural conditions for enhanced methionine production by Bacillus cereus S8

Study design: Study of the fermentation process in shake flask culture.

Place and Duration of Study: Department of Applied Microbiology and Brewing, Nnamdi Azikwe University, Awka, Nigeria between 2011 to 2012.

Methodology: The effects of medium/fermenter volume ratio, carbon and nitrogen sources, growth stimulators, vitamins and amino acid on methionine accumulation in the broth culture of Bacillus cereus S8 were investigated. The time course for methionine production was also studied.

Results: A 20% medium/fermenter volume ratio improved methionine yield. Glucose and ammonium sulphate at 6.0 and 1.0% respectively stimulated methionine accumulation by Bacillus cereus S8. Yeast extract, peptone, DL-leucine and all vitamins studied enhanced methionine production. A methionine yield of 3.23mg/ml was produced after 96h fermentation and at a pH of 6.90.

Conclusion: Improving the cultural conditions of Bacillus cereus S8 in submerged medium stimulated methionine increase

Open Access Original Research Article

Effect of Biochar on the Abundance of Soil Bacteria

T. F. Khan, S. M. Imamul Huq

Microbiology Research Journal International, Page 896-904
DOI: 10.9734/BMRJ/2014/9334

A pot experiment was conducted to study the effect of biochar on the abundance of soil bacteria and compare it with the source biomass. Seven different treatments and a control were used in the experimental set-up. Three different types of biomass were selected and three types of biochar were produced from them. Both the materials were applied to the soil at a rate of 5t/ha. All treatments were incubated for 30, 60 and 90 days. Cultural, microscopic and biochemical tests were carried out to identify the bacterial isolates in soils treated with biochar and its source biomass. Bacterial isolates identified in soil and in some of the biomasses before treatments were applied include Bacillus badius, Bacillus krulwichiae, Bacillus siralis, Bacillus sylvestris, Bacillus flexus, Aneurinibacillus aneurinilyticus and Bacillus thuringiensis while after incubation periods, seven new isolates were identified. This was true for the biomass treated soils where additional one to two isolates reappeared. Conversely, in the biochar treated soils, most of the isolates disappeared except Bacillus badius that survived in all soils till 90 days. Because of its tolerant nature, it was further investigated for cellulase enzyme activity. Interestingly, the isolate did not show any such activity. Conclusively, biochar application may exert negative effect on the distribution and proliferation of soil bacteria with possible effect on soil quality and crop production

Open Access Original Research Article

Seroprevalence of Hepatitis E Virus among Domestic Animals in Plateau State–Nigeria

Surajudeen A. Junaid, Samuel E. Agina, Kemi Jaiye

Microbiology Research Journal International, Page 924-934
DOI: 10.9734/BMRJ/2014/10203

Aim: This study was undertaken to determine the epidemiology, seroprevalence and associated risk factors, of Hepatitis E virus (HEV) among domestic animals.

Study Design: Cross sectional epidemiological survey.

Place and Duration: The study was carried out in three geographical zones of Plateau State, over a six month period from July to December, 2012.

Methodology: A total of 166 animal subjects were recruited into the study.

The animals studied were made up of pigs (67), goats (43), sheep (19) and cattle (37). Information was obtained from the animal subject handlers using interviewer administered questionnaire. Blood samples were collected and analyzed for HEV antibodies (IgG and IgM) using Enzyme-linked immunosorbent assay (ELISA) technique. Data obtained were analyzed using SPSS version 17.0 statistical software.

Results: Results revealed an overall prevalence of 24.1% (p<.001) with IgG and IgM accounting for 16.3% and 7.8% respectively. Goats recorded the highest prevalence with 37.2%, followed by Pigs with 32.8% and Sheep with 10.5%, but it is note-worthy that Cattle recorded 0% overall seropositivity. Statistical significant association was observed with regard to age (p=.04); animals <1 year old accounted for the highest seroprevalence (21.3%) and least among animals ≥2years old (7.7% OR 0.3; 95%CI0.1-1.1). Seropositivity tends to decreases with increase in age. A similar trend was observed with regard to IgM seropositivity. The significant associated risk factor was; frequency of waste disposal (p<.001) (IgM, OR 39.1; 95% CI 4.9-310.4; IgG, OR 19.9; 95% CI3.9-100.7). Animals that had been vaccinated against other diseases tend to exhibit the least seropositivity compared to animal subjects with no history of any form of vaccination.

Conclusion: Data suggest that HEV remains an under-recognized and significant public health issue in the study area, and prevalent among domestic animals, warranting further attention and research.  Preventive public health measures should be reinforced among all communities’ particularly domestic animals and a periodic monitoring system set up for control.

Open Access Original Research Article

Physico-chemical Quality Characteristics and Hazard Analysis Critical Control Points for the Production of Millet-based Kunun Zaki Obtained from Three Production Locations in Port Harcourt, Nigeria

C. N. Elele, S. N. Ibe, G. O. Abu

Microbiology Research Journal International, Page 935-948
DOI: 10.9734/BMRJ/2014/9156

Aim: To study the commercial preparation of Kunun zaki in three locations in Port Harcourt namely; Bori-camp, Rumuodomaya and Mgbuogba, to compare the microbiological, physico-chemical, and nutritional qualities of commercial and laboratory-prepared Kunun drinks and to establish the critical control points at various points of the production process.

Place and Duration of Study: Sample collection areas were Bori-camp, Rumuodomaya, Mgbuogba, and the Laboratory, between August and December 2012. Sample analysis was done in the Food and Industrial Microbiology Laboratory of University of Port Harcourt.

Methodology: At selected stages of preparation of Kunun zaki from Bori-camp, Rumuodomaya, Mgbuogba and the Laboratory, nine samples each were collected into sterile screw-capped 50cl bottles for analysis at the Food and Industrial Microbiology laboratory of University of Port Harcourt. Analyses carried out were to determine the microbial contaminants, proximate and physico-chemical parameters. Samples of fresh Kunun zaki were stored at 5ºC for three days for analysis on effect of storage.

Results: The microorganisms associated with freshly prepared Kunun zaki and that stored at refrigeration temperature (5ºC) for three days were Lactobacillus spp., Bacillus spp., Leuconostoc spp., Streptococcus spp., Micrococcus spp., Staphylococcus spp., Pseudomonas spp., Escherichia coli, Enterobacter spp., Penicillium spp., Mucor spp., Aspergillus spp., Rhizopus spp., Candida spp., and Saccharomyces spp. Freshly-prepared Kunun zaki had the highest coliform and staphylococci counts of 8.0x104 and 8.3x103cfu/ml respectively in Bori-camp preparation, while Laboratory-prepared Kunun zaki harboured none of these organisms. Total viable counts of commercially-prepared (Bori-camp, Rumuodomaya, and Mgbuogba) products ranged from 2.50x104 to 1.53x106cfu/ml, while Laboratory-prepared product was 6.0x104cfu/ml. Fungal counts of commercially-prepared Kunun zaki ranged from 2.5x103 to 1.36x105cfu/ml, while Laboratory-prepared Kunun zaki had fungal counts of 5.6x102cfu/ml. Yeasts were the main spoilage organisms which persisted at storage temperature of 5ºC for three days. The protein and carbohydrate contents, as well as calcium, zinc, copper, and manganese decreased after being stored for three days at 5ºC.

Conclusion: Therefore Kunun zaki could be safely consumed after storage at 5ºC for three days, if good manufacturing practices like the use of fresh non-moldy grains and spices, cooled boiled water for grain washing and steeping, sterile stainless steel containers, steam-sterilized grinder, and sterile screw-capped bottles for packaging be applied at all the production stages. Educating the producers on the hazards, critical control points (steeping, milling and packaging), and temperature maintenance for Kunun zaki preparation are important.