Open Access Original Research Article

Prevalence and Antibiogram of Gram-negative Bacteria Isolated from Well Water in Ula-Ubie Community, Ahoada West, Nigeria

R. R. Nrior, M. Okpokiri, N. P. Akani

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/mrji/2020/v30i230192

Antibiotic resistance has become a great global problem. Thus, it has emerged as a public health challenge. The antibiotic susceptibility pattern of bacteria in well water was characterized with a view of determining the level of resistance in the environment. Fifty well water samples were collected from ten different points in Ula-Ubie community, Ahoda, Rivers State for a period of five months. Standard microbiological methods were used to analyse the population and types of bacteria in the water while methods recommended by the American Public Health Association (APHA) was used to determine the physicochemical parameters of the samples. The antibiotic susceptibility profile of the bacterial isolates was carried out using the disc diffusion methods. The total heterotrophic bacteria of the water samples ranged from 0.93±0.46 to 2.02±1.06 log10 CFU/ml. The coliform counts ranged from 0.45±0.42 - 2.55±2.33 log10 CFU/ml, respectively. Despite the variations in the counts of the different bacterial population, there was no significant differences (P > 0.05) in the different well water samples. The physicochemical parameters except the pH were all within the permissible limits. Klebsiella spp, Pseudomonas spp, Serratia spp and Enterobacter spp were identified in the well water. The pH of the water stations ranged from 4.66 to 5.80. The temperature ranged from 24.0 to 24.7. The electrical conductivity, salinity, dissolved oxygen, total hardness, alkalinity, total suspended solids, biochemical oxygen demand, nitrate, chloride, calcium and magnesium ranged from 22.9 – 219, 0.03-0.13, 4.50-4.90, 5.00-22.0, ˂0.01-3.00, ˂0.01, 49.6-84.5, 1.00-17.4, 3.00-24.5, 4.25-12.9 and 0,722-1.55 respectively. The antibiotic susceptibility profile showed that all the isolates were resistant to ceftazidime and augmentin, whereas Enterobacter spp were the most resistant bacteria amongst other bacterial genera to the antibiotics. Meanwhile there is an existence of multi-drug resistance. Thus, the wells could be considered not potable due to the presence of these bacterial isolates and the level of antibiotic resistant. Proper sanitation and cleanliness of well should be encouraged.

Open Access Original Research Article

Clinico-microbilogical Profile of Chronic Osteomyelitis in a Tertiary Care Hospital of North India

Humaira Bashir, Asifa Nazir

Microbiology Research Journal International, Page 11-18
DOI: 10.9734/mrji/2020/v30i230193

Aims: Chronic osteomyelitis is an important clinical entity in patients with bone infections and is associated with great morbidity especially in developing countries. It is a persistent disease, difficult to treat and eradicate completely. Early identification and diagnosis of osteomyelitis has led to the improved management of osteomyelitis. This study was undertaken to determine the bacteriological profile of osteomyelitis and the antibiotic resistance pattern of various isolates obtained.

Study Design: It was a prospective cross-sectional hospital based study.

Place and Duration of Study: The present study was conducted in the Department of Microbiology, Government Medical College, Srinagar, from January 2019 to December 2019.

Methodology: In all, 208 patients with chronic osteomyelitis were documented during the study period. Clinical specimens like pus, pus swabs, sequestrum of bone and synovial fluid were taken and cultured aerobically .The samples were processed using standard microbiological techniques. Identification and antimicrobial susceptibility pattern of the bacterial isolates were done using the Vitek 2 (bioMerieux, France) system.

Results: A total of 208 samples were received out of which, 91 (43.75%) were positive by culture. Out of the 91 positive samples, Staphylococcus aureus 65 (64.2%) was the most commonly isolated pathogen and 67.6% of Staphylococcus aureus were MRSA. Other important organisms isolated included Acinetobacter sp (10.98%), E. coli (6.59%), Proteus mirabilis (4.3%), Pseudomonas aeruginosa (2.1%) and Klebsiella pneumoniae (2.1%). All the isolates of Staphylococcus aureus were resistant to Penicillin. However, Vancomycin resistance was not detected in any of the patients with MRSA. All Gram negative bacilli were sensitive to Colistin.

Conclusion: The wide range of causative organisms and degree of resistance to commonly used anti- microbials supports the importance of pus culture and provides important information to guide clinician’s choice of empirical antibiotics. Appropriate selection of antibiotic would help to treat the disease successfully and limit the emergence of drug resistant strains to prevent morbidity & mortality.

Open Access Original Research Article

PCR-RFLP of the Vitamin D Receptor Gene in Human Immunodeficiency Virus Patients Deficient in Vitamin D3 in Cote d’Ivoire

Lydie Boyvin, Aya Jeanne Armande Aké, Yapi Guillaume Yayé, Moumouni Faïza Alassani, Kipré Laurent Séri, Gnogbo Alexis Bahi, Louise Odile Moke-Bédji, Joseph Allico Djaman

Microbiology Research Journal International, Page 19-25
DOI: 10.9734/mrji/2020/v30i230194

Aims: This study was to identify mutations in patients’ vitamin D receptor (VDR) gene in Côte d'Ivoire, precisely in Human immunodeficiency virus (HIV) patients deficient in vitamin D3.

Methodology: Fifty (50) DNA extractions from peripheral blood mononuclear cells collected from HIV positive and vitamin D3 deficient patients were analyzed after verifying their integrity by quantification of genomic DNA and migration from agarose gel. The use of the restriction enzymes Dpn I, Bg III and Pst I made it possible to carry out the PCR-RLFP of the fragments Fok-1 in exon 2, Bsm-1 and Apa-1 in intron 8 and Taq-1 in exon 9.

Results: The analysis of the DNA fragments Fok-1 in exon 2 and Bsm-1 in intron 8 of the VDR gene from HIV positive patients deficient in vitamin D3 showed a significantly high prevalence of mutant genotype (100% and 98%) respectively p = 0.0001. Furthermore, in this study, a prevalence of 6% of mutant genotype was observed in Taq-1 of exon 9 of the VDR gene.

Conclusion: The high prevalence of mutant genotypes observed in the DNA fragments of Fok-1 in exon 2 and Bsm-1 in intron 8 of the VDR gene studied confirms the presence of mutations in the VDR gene of these patients. It would, therefore, be necessary to sequence the DNA fragments with mutations in order to identify the mutations that affect the VDR gene and that are responsible for the vitamin D3 deficiency observed in these patients.

Open Access Original Research Article

Staining Property of Alcoholic and Aqueous Hibiscus sabdariffa Extract in Demonstration of Selected Bacteria in Tissue Sections of Wistar Rats

S. Y. Ma’aruf, M. O. Mohammed, A. T. Muhammad, O. O. Okechi, R. I. Tsamiya, U. Abubakar, I. Mohammed, A. Umar, S. M. Sani, H. Kabir, B. A. Bello, S. Garba, S. D. Abubakar

Microbiology Research Journal International, Page 26-33
DOI: 10.9734/mrji/2020/v30i230195

Introduction: Dye is used for the artificial colouration of a substance to facilitate its examination by the use of the coloured organic molecule, a process called staining. Over the past many years, it has been observed that synthetic dyes have many disadvantages associated with them like toxicity and allergenicity. This study aimed to determine the staining effect of Hibiscus sabdariffa extracts of various concentration, various pH and duration in bacteria staining compared with Gram’s staining.

Methods: Standard Gram stains as control and both Hibiscus sabdariffa extracts (alcohol and boiled water) were used to stain inflammatory appendix tissue and lung tissue inoculated with bacteria using various concentration (5% and 10%), at various duration (30 seconds and 1 hour) and with change of pH, achieved by treating the extracts with ammonium hydroxide and glacial acetic acid. Each was used as a counterstain in bacterial stain (to replace Safranin/Neutral red) Safranin/Neutral red.

Results: All extracts after treatment were acidic but the change of pH was indirectly proportional to the staining ability of the extracts. The Hibiscus solutions gave the background, a brown colouration. And inflammatory cells were demonstrated better than the bacteria with aqueous Hibiscus solution strongly when compared with the alcoholic Hibiscus solution.

Conclusion: The results obtained indicate that Hibiscus solution has the potential for use in diagnostic bacteriology in formalin-fixed paraffin-embedded tissue section.

Open Access Original Research Article

Optimization of Some Nutritional Conditions and α- Ketoglutaric Acid Concentration as PGA Precursor for Maximizing PGA Production from Bacillus sp. 42 and Bacillus sonorensis 44

Rania F. Ahmed

Microbiology Research Journal International, Page 34-42
DOI: 10.9734/mrji/2020/v30i230196

Poly gamma glutamic acid is a biodegradable, water soluble and non-toxic edible biopolymer, PGA has nylon back bone similar structure and expressed as bio-nylon. Various bacterial strains produced PGA on of them Bacillus sp. such as B. subtilis, B. lichanformans and B. sonorensis. Polymer yield was affected with medium composition as nitrogen and carbon sources. The current experimental was carried out using shake flask technique for PGA production during 72 of fermentation. The highest biomass was achieved at glycerol media and glucose media for PGA yield and productivity being 2.31, 9.65 gl-1 and 0.134 gl-1h-1, respectively of B. sonorensis 44. Of nitrogen source, organic source (yeast extract) was higher PGA yield  and productivity than inorganic sources (NH4NO3) which reduced PGA yield about 28.7 and 36.02% of Bacillus sp. 42 and B. sonorensis 44, respectively. Production media supple-  mented with 0.5 and 0.75 gl-1 α-keto-glutaric acid increased PGA yield about 1.24fold for both Bacillus strains. Osmotic pressure of 2.55 MPa (3% NaCl) enhanced PGA yield about  1.18 and 1.24fold of Bacillus sp. 42 and B. sonorensis 44, respectively. Furthermore, the highest PGA was received using medium containing glucose and yeast extract (as C and N2 sources). α-keto-glutaric acid and osmotic potential has an induction effect for polymer accumulation.

Open Access Original Research Article

Comparison of Automated and Manual Methods for Antimicrobial Susceptibility Testing

Asifa Bhat, Dekyong Angmo, Shaista Nazir

Microbiology Research Journal International, Page 43-48
DOI: 10.9734/mrji/2020/v30i230197

Background: Carbapenems are considered the broadest-spectrum β-lactam agents and are often required for treatment of severe hospital-acquired infections caused by multidrug-resistant Gram-negative organisms. Minimum inhibitory concentrations (MICs) are important in diagnostic laboratories to confirm resistance of microorganisms to an antimicrobial agent and also to monitor the activity of new antimicrobial agents.

Aims and Objectives: To compare the MIC obtained by Broth Microdilution method (BMD) with that of Vitek-2(automated method) for recovered isolates of Klebsiella pneumoniae.

Materials and Methods: Prospective study conducted over a period of one year. It included all isolates of Klebsiella pneumoniae recovered from blood culture of the patients. The identification and antimicrobial susceptibility was done on Vitek-2.These Isolates were subjected to Microbroth dilution method for MIC determination.

Results: Out of the 55 meropenem resistant  isolates by vitek-2, 20(36.3%) had MIC of ≥256 µg/ml followed by 18(32.7%) isolates with a MIC of 128 µg/ml, followed by 11(20%) isolates with MIC of 64 µg/ml and 6(10.9%) isolates with MIC of 32 µg/ml. Also among 15 meropenem sensitive isolates by Vitek-2, 13(86.7%) had MIC of ≤0.5 µg/ml, followed by two (13.3%) isolates with MIC of 2 µg/ml. Results obtained by vitek 2 were compared with those from BMD(the reference method), which showed a 13.3% minor error rate and no major or very major error rate.

Conclusion: Overall, the Vitek 2 performance was comparable to that of BMD for testing a limited number of Klebsiella pneumoniae isolates.

Open Access Original Research Article

Preliminary Screening of Bacteria Isolated from Insects Living in Poultry for Antibiotic Production

Ifeoma Geraldine Okudo, Chinelo Ursula Umedum, Stephen Nnaemeka Ezekwueche, Chibuzo Christian Uba

Microbiology Research Journal International, Page 49-57
DOI: 10.9734/mrji/2020/v30i230198

Aim: This present study was conducted to isolate antibiotic producing bacteria from insects living in poultry.

Place and Duration of Study: Insects living in poultry were collected from poultry farms within Onitsha metropolis in Anambra State between April 2018 and September 2018.

Methodology: The gut of one hundred insects; (Musca domestica and Alphitobius diaperinus) were analyzed. The insects were dissected and emulsified in 10ml of peptone water. The dilutions were cultured on Nutrient agar and Blood agar  for 24 h. The bacterial isolates were characterized using  molecular identification. The DNA was extracted from the identified isolates and analyzed by 16S rRNA. In preliminary screening, the isolates were inoculated into Muller Hinton agar using agar plug. The promising isolate showing antagonism was subjected to submerged fermentation method and the secondary metabolites were extracted. Screening of the secondary metabolites extract was done using agar well diffusion. The minimum inhibitory concentration (MIC) of the secondary metabolite was determined using broth dilution method at different concentrations. The inhibitory activity of the organism was checked against four bacteria namely; Bacillus subtilis, Salmonella serovar typhi, Escherichia coli and Staphylococcus aureus.

Results: The sequence analysis revealed the strains to be Lysinibacillus macroides, Paealcaligenes hermetiae, Bordetella flabilis, Bacillus aerophilus, Klebsiella variicola. Lysinibacillus macroides showed antagonism against the test bacteria during the preliminary test. After fermentation, the secondary metabolite extracts from Lysinibacillus macroides exhibited antibacterial activities against Salmonella Serovar Typhi, Staphyloccus aureus and Bacillus subtilis at different concentrations.

Conclusion: The extracts from bacterial isolate; Lysinibacillus macroides exhibited antibacterial activities against Bacillus subtilis, Salmonella serovar typhi and Staphylococcus aureus. The extracts may serve as a new drug molecule produced from natural source when purified.