Open Access Original Research Article

Changes in Serum Levels of TNF-α & IL-4 among New, Under-treatment & MDR TB Patients

Nazish Fatima, Mohammad Shameem, Nabeela ., Haris M. Khan

Microbiology Research Journal International, Page 159-166
DOI: 10.9734/BMRJ/2015/14798

Introduction: Tuberculosis (TB) remains a significant public health problem with an estimated one-third of the world’s population being infected. Cytokines play a major role in protection against Mycobacterium tuberculosis infection and regulate the immune responses at cellular level. Most studies on cytokines during TB are from ‘in vitro’-stimulated lymphoid cells with few reports on in vivo plasma levels. This study was aimed to evaluate the levels of TNF-α & IL4 in new, under-treatment (UT) and multidrug resistant (MDR) pulmonary and extra-pulmonary cases.
Methodology: The study was conducted at the Department Of Microbiology, Jawaharlal Nehru Medical College, Aligarh Muslim University, Aligarh, India. Levels of TNF-α and IL-4 were measured in 76 serum samples from TB patients by ELISA kit (Diaclone France) along with 10 BCG vaccinated control. A complete clinical, radiological & treatment data was collected on questionnaire forms of each patients.
Results: TNF-α levels were elevated in new (P<0.05) and MDR cases (P<0.05), but not significantly for UT cases (P>0.05). TNF-α and IL-4 levels showed no significant variations according to site of involvement in pulmonary vs. extra-pulmonary TB cases.
Conclusion: An understanding of the development of this response may lead to insight into pathogenesis and novel therapies for TB.

Open Access Original Research Article

Antibiotic Susceptibility Profile of Bacteria Isolates from Some Fishponds in Niger Delta Region of Nigeria

O. E. Njoku, O. K. Agwa, A. A. Ibiene

Microbiology Research Journal International, Page 167-173
DOI: 10.9734/BMRJ/2015/15453

Aims: To determine the antibiotic susceptibility profile of bacterial isolates from concrete and earthen fish ponds.
Study Design: Comparative analysis.
Place and Duration of Study: The African Regional Aquaculture Center Allu (ARAC), Port Harcourt River State, from August 2013-Jauary 2014.
Methodology: Water samples from both concrete and earthen fish ponds were collected and analyzed at the environmental microbiology laboratory of the University of Port Harcourt. The bacterial isolates present in water samples were isolated using the spread plate technique, and identified using biochemical characteristics while the antibiotic susceptibility pattern of the isolates was obtained using the disc-diffusion method.
Results: A total of 94 bacterial isolates (47 from concrete and 47 from earthen ponds) comprising of the following genera Escherichia coliStaphylococcus sp., Aeromonas sp., Salmonella sp., Shigella sp., Pseudomonas sp., Vibrio sp., Proteus sp., Enterobacter sp., Serattia sp., Streptococcus sp. and Klebsiella sp., were identified. Data showed that all the isolates from both concrete and earthen ponds exhibited multiple drug resistance (MDR). All the isolates were 100% resistant to ceftazidime, cefuroxime, cefixime and cloxacillin in both ponds. The highest susceptibility of 100% was recorded with erythromycin in concrete ponds and 0% in earthen pond, ofloxacin 85% in concrete pond and 75% in earthen followed by gentamicine with 91% in concrete pond and 50% in earthen ponds. Among the isolates, Enterobacter species were resistance to all antibiotics tested (100%), followed by Proteus sp., Vibrio sp. and E. coli with 87.5% respectively while the least resistance was found in Pseudomonas sp. (50%).
Conclusion: In conclusion, the prevalence of antimicrobial resistance in the pond water is a serious health concern since the microbial flora of a fish is the reflection of its aqueous environment. This implies that antibiotic resistant bacteria present in the pond water will also be present in the cultivated fish and fish product. The multiple antibiotic resistances among the pond water pathogens may further complicate the clinical management of diseases caused by the consumption of fish cultivated in these ponds.

Open Access Original Research Article

Relationship between Gingivitis and Autism in Children: A Matched Case-control Study

Archana Singh Sikarwar, Abhishek Parolia, Fransazellea Anak Robert Runai, Ankur Barua

Microbiology Research Journal International, Page 174-179
DOI: 10.9734/BMRJ/2015/14108

Autism is a neurodevelopmental disorder diagnosed in early childhood. Studies have shown that autistic children have poorer gingival health and hence chances of higher incidence of gingivitis may occur in autistic than non-autistic children. Gingivitis is characterized by gingival inflammation without loss of attachment due to continuous exposure to dental plaque. Studies on the gingival health of autistic children in Malaysia are lacking. Hence, this study aims to evaluate and compare the gingival health status of autistic and non-autistic children.
Materials and Methods: A total of 86 children were recruited and an oral assessment was performed on each child to assess the degree of calculus, plaque accumulation and gingival inflammation using the calculus, plaque and gingival indices respectively. Pooled dental plaque (supragingival margins and gingival crevices of the buccal surface of the anterior maxillary molar and lingual of mandibular molar teeth) samples were collected and transferred to Stuart transport media. Samples were diluted and 100 µl aliquots were cultured on Columbia agar with 5% sheep blood. Plates were incubated in an anaerobic environment at 37ºC for 7 days. Colonies were Gram stained and examined for cellular morphology followed by indole and catalase test. Bacterial identification was done using the Rapid ID 32 A kit. Kolmogorov-Smirnov, Chi-square, Mann-Whitney U, Odds ratio and Spearman’s correlation analyses were used to analyze the data.
Results: The prevalence of dental calculus was significantly elevated in the autistic group with 43 sample size in each group (p = 0.001). However, the prevalence of gingivitis and plaque were not significantly different between the two groups. The presence of plaque (p = 0.001) and calculus (p = 0.004) were significantly associated with the development of gingivitis.
Conclusion: While the autistic children had significantly more calculus, the occurrence of gingivitis and plaque were similar between autistic and control groups. Measures to control dental plaque and calculus among the autistic children should be improved to prevent gingivitis.

Open Access Original Research Article

Evaluating the Effect of Gamma Irradiation and Steam Sterilization on the Survival and Growth of Composted Sawdust Fungi in Ghana

N. K. Kortei, G. T. Odamtten, M. Obodai, V. Appiah, M. Wiafe-Kwagyan

Microbiology Research Journal International, Page 180-192
DOI: 10.9734/BMRJ/2015/16521

The growth and survival of some fungi associated with composted and pretreated sawdust particularly for mushroom cultivation were investigated on two growth media; Cooke’s and Oxytetracycline Glucose Yeast Extract (OGYE). Some fungi were isolated during the composting of sawdust over a period of 28 days as well as after pretreatment with gamma irradiation doses of 5, 10, 15, 20, 25 and 32 kGy and moist heat of 100±2ºC for 2.5 hours. Fungal counts ranged 4.72- 5.77 log10 CFU/g and 3.4- 4.1 log10 CFU/g respectively for both media. Both pretreatment methods effectively reduced (p<0.05) fungal counts by an average of 1.48 (irradiation) and 2.22 (steam) log-cycle reductions on OGYE while there was an average of 3.13 (irradiation) and 1.10 (steam) log-cycle reduction on Cooke’s. Corresponding radiation sensitivities (D10 values) of 5.94±2.06 kGy and 5.64±1.12 kGy were recorded for fungi on both media respectively. Five species belonging to three genera were isolated on OGYE and among the fungi were Aspergillus niger, Aspergillus fumigatus, Aspergillus ustus, Mucor racemosus and Rhizopus stolonifer. Ten species belonging to four genera were also isolated on Cooke’s medium included Aspergillus niger, Aspergillus fumigatus, Aspergillus flavus, Aspergillus ustus, Aspergillus terreus, Aspergillus parasiticus, Aspergillus alutaceus, Rhizopus stolonifer, Fusarium oxysporum and Mucor racemosus. There was an observed reduction (p<0.05) in species diversity after pretreatment. The presence of these microorganisms on sawdust also has serious repercussions on animal and human health.

Open Access Original Research Article

Genetic Diversity Study of Locally Isolated Bacillus thuringiensis Strains from Kuwait Using Random Amplified Polymorphic DNA Analysis

Jafar A. Qasem, Ebtesam M. Al-Ali, Salwa A. Al-Mouqati, Yousif A. Al-Shayji

Microbiology Research Journal International, Page 193-201
DOI: 10.9734/BMRJ/2015/17009

Introduction: Bacillus thuringiensis (Bt) is significant bacteria in the field of insect biological control due to their insecticidal properties and its importance in agriculture.
Aim: The objective of this study was to analyze genetic variation of 15 native isolates of Bacillus thuringiensis and four reference strains (total 19) using the random amplified polymorphic DNA (RAPD) technique, this allowed the analysis the genetic diversity of this species in the microbial populations from different soil samples of Kuwait.
Methodology: We isolated 109 Bacillus thuringiensis strains, out of which, 15 strains were subspeciesthuringiensis, using culture and serological method. A rapid typing method of Bacillus thuringiensis local isolates from Kuwait soil was established using the RAPD technique. A single decamer primer was used to study and characterize Bacillus Spp, Bacillus thuringiensis var. thuringiensis, to differentiate the isolated species. Based on RAPD pattern, data were subjected to cluster analysis using Alpha Ease software.
Results: We found three groups each, with two strains that had a similar pattern of DNA and one group that had three subspecies that have a similar DNA pattern. The rest of the eight isolates each had a unique pattern of DNA. These isolates were classified according to the same chemical and physical characterization, but were different genetically.
Conclusion: In conclusion, using molecular methods for comparison of genomic DNA between different bacterial species of the same genus is a good measurement of genetic relations between different species, which could lead to discovery of new species unique to the local environment.

Open Access Original Research Article

Molecular Identification of mecA gene in Methicillin-Resistant Staphylococcus aureus Isolated from the Hospitalized Patients in Teaching Hospitals of Ahvaz, Iran

Mojtaba Moosavian, Najmeh Parhizgari

Microbiology Research Journal International, Page 202-209
DOI: 10.9734/BMRJ/2015/14296

Aims: Staphylococcus aureus is a causative agent of nosocomial infections. Methicillin is one of the most important antibiotics that is used in treatment of S. aureus infections; however, resistance to this antibiotic has occured in recent years. The aim of this study was detection of mecA gene in methicillin-resistant S. aureus (MRSA) strains which were isolated from the hospitalized patients.
Methodology: In this study, 255 Staphylococci isolates were collected from the patients with infection in three teaching hospitals. These strains were isolated from different specimens and identified using microscopic and standard biochemical tests. S. aureus strains phenotypically resistant to oxacillin, were screened after determination of drug resistance patterns against 9 antibiotics by disk diffusion method based on CLSI guidelines. Then, DNA was extracted from phenotypic MRSA strains and finally mecA gene was amplified by PCR.
Results: Out of 255 Staphylococci isolates collected from the patients with infection, 180 isolates were confirmed as S. aureus. The results of the antibiogram test revealed the highest and lowest rate of resistance against gentamicin and nitrofurantoin, respectively. These results also showed that out of 180 isolates of S. aureus, 59 isolates (32.7%) were identified phenotypically as MRSA and mecA gene was confirmed in 58 of them by PCR method.
Conclusion: The results of our study showed high prevalence of HA-MRSA isolates among of the examined patients, as nearly one third of these isolates were resistant to methicillin. So, for prevention of the spread of Health care- associated MRSA infection, it is necessary to augument control measures of hospitals and it is recommended that the patients with staphylococcal infection be treated only after verification of drug sensitivity of isolates by clinical laboratories.