Open Access Original Research Article

Microorganisms Associated with Biogas Production Using Vegetable (Telfairia occidentalis) Wastes, Banana Peel and Pig Dung as Substrates

B. E. Asikong, S. O. Idire, D. R. Tiku

Microbiology Research Journal International, Page 1-12
DOI: 10.9734/BMRJ/2016/28294

The research study was aimed at investigating microorganisms associated with biogas production using vegetable (Telfairia occidentalis) wastes, banana peel, and pig-dung as substrates. Marian market, Watt market and University of Calabar pig farm were randomly sampled within Calabar metropolis for collection of samples. The study was completed within a period of six month. Standard microbiological methods and anaerobic biodigesters were used to screen the isolates and the wastes substrate for biogas production. Analysis revealed that the temperature of raw substrates ranged between 21°C and 39°C while the pH varied between 6.10 and 7.21 during digestion. Highest mean bacterial counts was 8.87±3x106 cfu/g and fungal count of 5.67±105 cfu/g were obtained in the combined substrates of banana peel, vegetable waste and pig dung (BP + VW + PD) before digestion, as compared to mean bacterial counts of 8.62±1.4x10fu/g and fungal counts of 5.55±1.7x105 cf/g obtained during digestion. Anaerobic bacteria isolated were identified as, Pseudomonas sp, Escherichia coli, Bacillus sp, Salmonella sp, Staphylococcus aureus, Serratia sp, Shigella sp, Micrococcus sp, Proteus vulgans, Citrobacter sp and Klebsiella sp, while fungi isolated were identified as Fusarium sp, Mucor sp and Penicillium sp. Methanogenic bacteria isolated were identified as Methanothrix sochngenii, Methanococcoides methylutens and Methanoculles bourgense. The volume of biogas produced and the percentage methane yield varied signficantly (p<0.05) between the substrate treatments and the digestion internals (days). However, the study has shown that the role of methanogens and other complementing bacteria and fungi in biogas production is indispensable.

Open Access Original Research Article

Detection and Molecular Characterization of ESBLs in E. coli Isolates from a Tertiary Care Hospital in North India with Special Attention to CTX-M-27

Nitika Anand, Ashish K. Asthana, Molly Madan

Microbiology Research Journal International, Page 1-7
DOI: 10.9734/BMRJ/2016/27860

Background: Extended spectrum beta-lactamases are the main cause of resistance to beta lactam antibiotics in members of Enterobacteriaceae. ESBL associated infections are on a rise worldwide and have become a serious public health problem. We aimed to investigate the molecular epidemiology of ESBL producing E. coli isolates recovered from various clinical specimens at a tertiary care hospital and to determine the antibiotic sensitivity profile of ESBL positive isolates.

Methodology: A total of 300 isolates of E. coli were collected from various clinical specimens between the study period of 2011 to 2014. Antimicrobial susceptibility testing was done. ESBL detection was carried out by CLSI Phenotypic confirmatory method. Molecular typing of ESBLs was performed by uniplex PCR among 100 ESBL isolates. The bla CTX-M strains were genotyped by sequencing of PCR product. Nucleotide sequences were submitted to Gen Bank and accession numbers were obtained. 

Results: 61% isolates were found to be ESBL producers. ESBL and non-ESBL producers compared among in- and out-patients gave statistically significant result (P value=0.002). All ESBL isolates (100%) were sensitive to imipenem. Overall 93.9% ESBL producers and 67.5% non-ESBLs were Multi Drug Resistant (Resistance to 3 or more class of antibiotics). The difference was statistically significant (P value=0.001). Majority of the typeable isolates harboured two or more ESBL genes (52%). Sequencing was done for 10  randomly selected  blaCTX-M PCR products and majority (90%) were identified as CTXM-15  belonging to CTX-M Cluster-1 while 1 0f 10 (10%)  was identified as CTX-M- 27 belonging to CTX-M Cluster-9 on blast  analysis. Deduced nucleotide sequences were submitted to Gen Bank. The accession numbers obtained from Gen Bank are KU946005-KU946009.

Conclusion: Our study shows high ESBL occurrence among E.coli isolates and highlights the incidence CTX-M-27 for the first time from North India.

Open Access Original Research Article

Prevalence and Risk Factors Associated with Intestinal Parasitic Infection among Patients in Taiz City, Yemen

Talal AL-Harazi

Microbiology Research Journal International, Page 1-7
DOI: 10.9734/BMRJ/2016/28317

Aims: To determine the prevalence and associated risk factors of intestinal parasites among patients in Taiz city.

Study Design: A cross-sectional descriptive study.

Place and Duration of Study: This study was carried out on patients visiting general and hospitals in Taiz, Yemen during April to September 2014.

Methodology: A total of 330 stool samples were collected from patients and analyzed by direct wet mount and formal ether concentration techniques. Furthermore, sociodemographic data were collected by using a standardized questionnaire.

Results: The overall prevalence of intestinal parasitic infections was 38.2%. The most predominant parasites found was Entamoeba histolytica/dispar (20.6%) followed by Giardia duodenalis (12.7%), respectively. Other parasites detected included Ascaris lumbricoides (3%), Hymenolepis nana (0.9%) and Schistosoma mansoni (0.9%). Multivariate analysis confirmed that drinking untreated water, not washing hands after defecation and contact with animals was a significant risk factor with parasitic infections.

Conclusion: The findings of this study indicated that intestinal parasitic infections are still a public health problem in Yemen. Statistical analysis indicated that low personal hygiene, lack of access to potable water and contact with animals were important predictors for intestinal parasitic infections. Hence, improving the knowledge on local risk factors such as contact with domestic animal, health status and personal hygiene is warranted.

Open Access Original Research Article

Brucella Prevalence in Goats and Farmers’ Awareness and Practices towards Brucella Infection in Giwa Area of Kaduna State Nigeria

R. Dogo, B. V. Maikai, J. A. Musa, J. Q. Tizhe

Microbiology Research Journal International, Page 1-12
DOI: 10.9734/mrji/2016/v16i35253

Aims: To detect and determine the prevalence of Brucella antibodies in goats and farmers’ awareness and practices towards Brucella infection in Kaduna State, Nigeria.

Experimental Design: A cross sectional study was used in this research.

Place and Duration of Study: Giwa area in Kaduna State, Nigeria. The study was conducted from July, 2014 to June, 2015.

Methodology: Two hundred and eighty serum and 113 milk samples (from lactating Does) were collected from goats in Giwa area of Kaduna State. Of the six districts in the area, 52 samples were collected from 10 households in Kakangi, 45 from 9 households in Giwa, 43 from 8 households in Tsibiri, 43 from 8 households in Gangara, 56 from 10 households in Yakawada, 41 from 9 households in Danmahawayi. Open and close ended questionnaires were administered to the farmers in form of interview using local dialect, to obtain information on their goats, such as age, sex, management practices, pregnancy, abortion history, and other reproductive problems such as retention of placenta in goats. Rose Bengal Plate Test (RBPT), Competitive Enzyme Linked-Immunosorbent Assay (cELISA) and Milk Ring Test (MRT) were used to detect Brucella antibodies. Open and close ended questions were administered to the farmers in form of interview to obtain information on their goats and to determine their knowledge and practices with respect to Brucella infection. Chi-square (χ2), Fisher’s exact tests and Odds ratio were used to test for association between categorical variables. P-value less than 0.05 (P<0.05) was considered statistically significant.

Results: A prevalence of 8.2%, 2.5% and 38.1% were obtained using RBPT, cELISA and MRT respectively. There was a significant association (P<0.037) between the districts and prevalence of Brucella antibodies in milk. The prevalence of Brucella antibodies was higher (10.9%) in goats from households where the farmers had no knowledge of the disease than those who had knowledge of the disease (5.9%). Those flocks with history of abortion had a lower prevalence (6.3%) as compared to those without abortion (12.5%). Detection of Brucella antibodies were higher (13.8%) in flocks that recorded abortion during the late gestation period than the early (5.0%) and mid (12.3%) gestation periods. Thirty-nine percent of these farmers used polythene bags on their hands as protective covering when handling aborted fetuses and other vaginal discharges, while 56.1% discard aborted materials freely into the environment. There was no statistical association between the prevalence of Brucella antibodies and farmers’ knowledge of the disease, abortion history and gestation period at the time of abortion.

Conclusion: This study has demonstrated that goats in Kaduna State harbour antibodies to Brucella and the farmers’ awareness and practices towards Brucella infection is insufficient. There is a need to further enlighten the farmers on the zoonotic implication of brucellosis, Brucella infection prevention and control.

Open Access Original Research Article

Biodegradation of Pyrene Using Bacillus sp.C7 Isolated from Coal Deposited Soil

M. A. Karale, T. A. Kadam, H. J. Bhosale, K. P. Maske

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/BMRJ/2016/27993

Aims: To evaluate the efficiency of bacterial isolates in pyrene degradation.

Place and Duration of Study: The study was carried out at School of Life Sciences, S.R.T.M. University, Nanded, (M.S.), India during June 2015 and March 2016.

Methodology: Coal deposited soil was collected and used for isolation of pyrene degrading bacteria. The isolation of bacteria was carried out following three successive enrichments of soil sample in Bushnell Haas broth supplemented with 200, 400 and 1000 mg/L pyrene respectively. The morphologically distinct 10 bacterial isolates obtained on Bushnell Haas agar medium supplemented with 1000 mg/L pyrene as sole carbon and energy source were screened for pyrene degradation ability by DCPIP assay. The efficient pyrene degrading Bacillus sp. C7 was selected and used for further studies. The parameters in terms of pyrene concentration, pH of the medium, temperature, incubation period and suitable surfactant, carbon and nitrogen source were optimized using OVaT approach.

Results: The efficient pyrene degrading strain C7 was selected and identified on the basis of morphological and biochemical characterization as Bacillus sp. C7. The strain was able to catabolize 81.53% of pyrene (1000 mg/L) within 20 days of incubation. The strain showed highest pyrene degradation at 1000 mg/L, 30°C, pH 7.0, after 20 days of incubation and in the presence of tween-80, glucose as co-metabolite and beef extract as nitrogen source.

Conclusion: The study identified a pyrene degrading soil bacterium Bacillus sp. C7. The optimization approach used in the study revealed tween-80, glucose and beef extract are principle components for pyrene degradation. Overall 5.17% rise over control in pyrene degradation was observed after optimization studies.

Open Access Short Research Article

Extended Spectrum β-lactamases in Clinical Isolates of Escherichia coli and Enterobacter cloacae Collected from Nablus District - Palestine

Motasem Al-Masri, Nael Abu-Hasan, Maha M. Jouhari

Microbiology Research Journal International, Page 1-7
DOI: 10.9734/BMRJ/2016/27892

Aim: To determine the prevalence of extended-spectrum β-lactamases (ESBLs) among clinical isolates of Escherichia coli (E. coli) and Enterobacter cloacae (E. cloacae) in Nablus district.

Methodology: In this prospective study carried out at An-Najah National University, a total of 161 bacterial isolates were collected during a12-month period in Nablus district in Palestine. To detect ESBLs, the isolates were examined by combination disc method. PCR was used to detect blaCTX-M, blaTEM and blaSHV genes in 32 representative ESBL-producer E. coli isolates.

Results: Using combination disc method, ESBL enzymes were detected in 73 out 153 (47.7%) E. coli and in 1 out of 8 (12.5%) E. cloacae isolates. No significant association of ESBL-producer E. coli was observed with types of collected specimens, gender, hospital ward, outpatient, or medical source. Among 32 representative E. coli ESBL-positive, blaCTX-M, blaTEM and blaSHV genes were detected in 30 (93.8%), 2 (6.3%) and 1 (3.1%), respectively. Two new variants of ESBLs (PALTEM137b and PALSHV-2a') were identified.  A unique E. cloacae isolate expressing inducible class C B-lactamase was also detected.

Conclusions: In Nablus region, high frequencies of ESBLs were found among E. coli bacteria isolated from outpatients and inpatients. blaCTX-M is the predominant gene among ESBL producers. New variants of ESBLs were found.