Open Access Original Research Article

Seroprevalence of IgM Antibody in Atypical Pneumonia Causing Pathogens by Pneumoslide, IFA

Puneeta Singh, Shalabh Malik, Vandana Lal

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/mrji/2020/v30i630223

Background: Atypical bacterial and viral pathogens play an important role in atypical pneumonia are responsible for one of the leading causes of morbidity and mortality, particularly in developing countries.

Objective: The purpose of this study to determine the prevalence of bacterial and viral pathogens causing acute atypical pneumonia in different age groups and seasonality patterns of prevalence in India.

Methods: This retrospective study was conducted on 680 samples tested during December 2018 to August 2019, performed at Microbiology department of Dr. Lal Path Labs. Serum samples were used for Pneumoslide IgM test diagnose 9 Atypical bacterial & viral pathogens: Legionella pneumophila (LP), Mycoplasma pneumoniae (MP), Coxiella burnetti (COX), Chlamydophila pneumonia (CP) Adenovirus (ADV), Respiratory syncytial virus (RSV) Influenza A (INFA), Influenza B (INFB), Parainfluenza serotypes 1,2 &3(PIVs).

Results: Of a total 477(70.1%) samples were positive for atypical pneumonia pathogens. Atypical pneumonia was seen in extremes of age ie: <=5 years and >60 elderly adults without much of a gender bias. Co infections was seen in 62.1%. Legionella pneumophila (42.5%) was the dominant pathogen followed by Influenza B (41.7%) Mycoplasma pneumoniae (33.4%), Parainfluenza serotypes 1,2 &3 (29.4%) respectively. Atypical pneumonia has a spring predominance that is peaking in March.

Conclusion: Among six predominant atypical pathogens, Legionella pneumophila and Influenza B was most predominant pathogens, as a causative agent of atypical pneumonia followed by Mycoplasma pneumoniae seen mostly in young (0-5 years) comparison to all age groups. Hence, Pneumoslide IgM as a multi panel test needed to ensure initiation of targeted therapy. Pneumoslide IgM, by IFA is a rapid, cost effective easy to identify & classify atypical pneumonia causing pathogens.

Open Access Original Research Article

Molecular Identification of Virulence Genes of Escherichia coli Isolated from Cow Milk and Its Products in Abuja, Nigeria

E. C. Okechukwu, E. U. Amuta, G. M. Gberikon, N. Chima, B. Yakubu, J. C. Igwe, M. Njoku

Microbiology Research Journal International, Page 11-18
DOI: 10.9734/mrji/2020/v30i630226

Shiga toxin-producing Escherichia coli have been identified as an emerging foodborne pathogen which portends serious risk to human health. Cow milk and its products are potential sources of shiga toxin-producing Escherichia coli. A relatively small number from the family of shiga toxin-producing Escherichia coli are pathogenic. It becomes necessary that Cow milk and milk products are regularly screened for the presence of virulence genes in microbes. The study aimed to genetically determine the presence of virulence genes that are characteristic of Enterohaemorrhagic E. coli in 600 milk samples. The E. coli isolates were recovered from the milk samples (n=35), biochemically examined and genetically screened for virulence genes by multiplex Polymerase Chain Reaction (PCR). The results of the molecular profiling revealed that (stx2) was detected in 17(60.7%), (hlyA) 11(39.3%) and eae genes 8(28.6%) of the E. coli isolates respectively, while (stx1) was not detected. The results indicated a high prevalence of virulent shiga toxin-producing Escherichia coli in the milk samples. Priority attention should be given to this microbe as it will demand stringent steps in the detection given that they are known to be rigorous in identification.

Open Access Original Research Article

Microbial Quality of Some Leafy Vegetables Sold in Iree

O. O. Efunwole, T. A. Ihum, O. R. Adebayo, O. A. Ajewole, D. O. Alaje

Microbiology Research Journal International, Page 19-22
DOI: 10.9734/mrji/2020/v30i630227

This study investigates the microbial quality of some common leafy vegetables sold in Iree. Three common leafy vegetables were used which were Amaranthus viridis (‘Tete’), Corchorus olito rus (‘Ewedu’) and Talinum triangulare (‘Gbure’). The samples were bought from the Iree main market and packaged in a sterilized aluminium covered  plates and were transferred immediately into the laboratory where they were examined for Total viable counts, coliform counts and Total fungal counts. The average total viable counts was 2.2 x 105cfu/mL for Amaranthus viridis, 2.3 x 105 cfu/mL for Corchorus olitorius and 1.9 x 105 for Talinum triangulare. Seven bacteria belonging to five genera and three fungal Spp. were isolated, which include, Staphylococcus aureus, Bacillus subtilis, Bacillus cereus, Salmonella enteritidis, Enterobacter aerogenes, Eschericia coli, Micrococcus luteus, Aspergillus niger, Aspergillus flavus, Penicilium italicum respectively. Staphylococcus (26%) was the most predominantly isolated followed by Bacillus subtilis (13%), Eschericia coli (12%), Streptococcus and Micrococcus (11%), Enterobacteriacea (9%), fungal (10%) and Bacillus cereus was the least (8%). These results showed that agricultural practices on these vegetables, like transportation, irrigation and even fertilizer application method could pose risks to the consumers, it is therefore advised that proper cooking of these leafy vegetables by boiling in 100°C boiling water, should strictly be adhered to before consumption not just mere blanching.

Open Access Original Research Article

Phytochemical Analysis, In vitro Antibacterial Activity and Rate of Kill of Different Fractions of Dacryodes edulis Leaf

R. E. Hassan-Olajokun, A. M. Deji-Agboola, O. O. Olasunkanmi, T. A. Banjo, O. Olaniran, A. E. Awoyeni, O. M. Ajayi

Microbiology Research Journal International, Page 23-35
DOI: 10.9734/mrji/2020/v30i630228

Dacryodes edulis is a fruit tree attaining a height of 18–40 meters mostly grown around the house and rarely in the forest. It has a relatively short trunk and a deep, dense crown and it is native to Africa hence often called African pear. The leaves are a compound with 5-8 pairs of leaflets. Acetone extract/solvent fractions of the leaves of Dacryodes edulis were used. The in vitro antibacterial activities and rate of kill of different fractions were investigated. The phytochemical screening was done by some chemical tests, antibacterial activity by agar well diffusion method and rate of kill was carried out on S. aureus and E. coli organisms. The fractions exhibited antibacterial activities with zones of inhibition ranged between 20 and 30 mm by Aqueous (AQU) fraction while Dichloromethane (DCM) fraction ranged between 22 and 32 mm. The standard antibiotics, streptomycin ranged between 10 and 20 mm and ampicillin between 11 and 27 mm. The Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) of both fractions had range values between 0.78 and 6.25 mg/mL. The phytochemical screening revealed the presence of alkaloids, tannins, saponins, flavonoids, terpenoid and glycoside. Overall, the two fractions had better activities than the standard antibiotics used. The time kill assay showed that the percentage of the cells killed increased with increasing concentrations of the fractions, as well as, contact time intervals. The AQU fraction killed 100% of Escherichia coli and Staphylococcus aureus cells at concentration of 4.68 mg/mL (3 X MIC) within 120 min while DCM fraction killed 100% of Escherichia coli cells within 90 min at a concentration of 4.68 mg/mL and killed 100% of S. aureus cells within 120 min at a concentration of 3.12 mg/mL (2X MIC). In conclusion, D. edulis leaf fraction has a broad spectrum antibacterial activity, with the AQU and DCM fractions being bactericidal as exemplified by the killing rate and MIC index of 2 (ratio of MBC/MIC) for both fractions.

Open Access Original Research Article

High Prevalence of Antibiotic Resistant Escherichia coli Serotype O157: H7 among Pastoral Communities in Rural Uganda

Abdul Walusansa, Jacob S. Iramiot, Christine F. Najjuka, Dickson Aruhomukama, Hussein Kafeero Mukasa, Henry Kajumbula, Benon B. Asiimwe

Microbiology Research Journal International, Page 36-43
DOI: 10.9734/mrji/2020/v30i630230

Background: Non-prescribed use of antimicrobials in Agriculture incurs a transfer risk of resistant pathogens to humans, complicating treatment. The aim of this study was to determine the potential of Zoonotic E. coli to serve as drivers of antimicrobial resistance (AMR) among animals and humans in pastoralist communities in Kasese district, so as to protect the community.

Materials and Methods: A laboratory based cross-sectional study was done using archived E. coli isolates previously obtained from humans in pastoralist communities of Kasese district, Uganda. Recovery of the isolates was done by conventional culture, and Identification by biochemical methods, serotyping and PCR. Kirby Bauer disc diffusion method was adopted for AMR profiling. Isolates were screened for resistance mechanisms including Extended Spectrum β-lactamase (ESBL), Carbapenemases and AmpC production using disc diffusion based methods.

Results: The prevalence of Enterohemorrhagic E. coli (EHEC) was 16% (28/180). These EHEC isolates belonged to phylogroups; B1 (94%, 26/28), B2 (3%, 1/28) and A (3%, 1/28). All the 28 EHEC isolates possessed the virulence gene stx1, 26 of the 28 EHEC isolates contained the virulence gene stx2e, but none of the 28 possessed the virulence gene stx2. Highest resistance was seen to Cotrimoxazole (89%, 25/28), Tetracycline (71%, 20/28), Ampicillin (65%, 18/28) and Nitrofurantoin (28%, 8/28), these are the most commonly used antimicrobials in the agricultural sector in Uganda. Minimal resistance was observed to the antimicrobials that are commonly used in human medicine especially β-lactams, β-lactam+inhibitors and Carbapenems. Of the 28 zoonotic E. coli isolates, 17%, (5/28) were ESBL positive and among these 1 (3%, 1/28) was a Carbapenemase producer.

Conclusion: There is a high prevalence of highly pathogenic, drug resistant E. coli O157:H7 among humans in pastoralist communities in Uganda. We suspect that these were acquired from animals because they mostly contained vero toxin gene vt2e which is animal specific, predominantly swine. Also majority of these EHEC isolates belonged to phylo-group B1 which has been documented to inhabit domestic animals. We recommend future studies to investigate relatedness of drug resistant isolates from humans and animals to ascertain the zoonotic spread of resistant enterohemorrhagic E. coli in pastoralist communities.

Open Access Original Research Article

Polycyclic Aromatic Hydrocarbon (PAH) Degrading Potential of Bacteria Isolated from Iko River Sediment

Stephen Akpan, Senyene Umana, Solomon Etuk

Microbiology Research Journal International, Page 44-54
DOI: 10.9734/mrji/2020/v30i630231

Polycyclic aromatic hydrocarbon (PAH) degrading potential of bacteria isolated from Iko River sediment, Akwa Ibom State, Nigeria was investigated. The mean total heterotrophic bacteria obtained from the sediment samples was 6.4 × 10 cfu/g while 9.8 x 103 cfu/g hydrocarbon utilizing bacteria was recorded. Preliminary screening of the hydrocarbonoclastic bacterial isolates revealed that among the 12 bacterial isolates, Pseudomonas aeruginosa, Bacillus subtilis, Alcaligenes sp exhibited the strongest ability to utilize crude oil. The result also revealed Bacillus subtilis and Pseudomonas aeruginosa as the best PAH degraders. A higher microbial cells count in the 2-ring PAH (naphthalene) supplemented –MSM was recorded. The levels of attenuance however varied with the test organism and were accompanied by fluctuations but decreasing pH levels and slight changes in temperature of the culture medium. In vitro degradation study carried out for the 21 days showed that the degradation of PAH when augmented with Pseudomonas aeruginosa and Bacillus subtilis respectively was faster than when un-augmented. The PAH content was reduced by Pseudomonas aeruginosa and Bacillus subtilis from 10.42 to 9.03 and 9.56 respectively. The hydrocarbon degradation by augmented cultures of Pseudomonas aeruginosa and Bacillus subtilis implies that bioaugmentation can be harnessed for bioremediation purposes.

Open Access Original Research Article

First Detection of the Plasmid-Mediated Quinolone Resistance Determinants qnrB, qnrS and aac(6’)-Ib-cr in Extended Spectrum Beta-Lactamases-producing Klebsiella pneumonia in Bouaké, Côte d’Ivoire

Marie N. Tuo, Augustin E. Anoh, Zéphirin O. Wayoro, Baba Coulibaly, Pacome Monemo, Nadine F. Wohi, Eloise Mueller- Schulte, Simon-Pierre A. N’guetta, Sören L. Becker, Chantal Akoua- Koffi

Microbiology Research Journal International, Page 55-67
DOI: 10.9734/mrji/2020/v30i630232

Aims: The aims of the present study were to investigate the presence of Plasmid-Mediated Quinolone Resistance (PMQR) determinants and the association of these determinants with Extended Spectrum Beta-Lactamases (ESBLs) genes in ESBL-producing Klebsiella pneumoniae isolates from Teaching Hospital of Bouaké, Côte d’Ivoire.

Study Design: It is a retrospective study.

Place of Study: Bacteriology-Virology Laboratory of Teaching Hospital, Bouaké, Côte d'Ivoire.

Methodology: From January 2015 to December 2016, 96 ESBL-producing Klebsiella pneumoniae isolates were collected from several specimens. Antimicrobial susceptibility of isolates was tested using the standard disk-diffusion method on Mueller-Hinton and interpretation according to recommendations of the 2017 EUCAST. These isolates analyzed for the detection of ESBL (blaCTX-M, blaTEM and blaSHV) and PMQR genes (aac(6’)-Ib-cr, qnrB and qnrS) using simplex PCR.

Results: Of the 96 ESBL-producing strains, 85 (88.55%) harbored at least one of the ESBL genes tested. Out of the 85 strains encoding ESBL genes, 96.47% carried blaCTX-M and 92.94% blaSHV and blaTEM genes. Eighty nine (89.6%) of the 96 ESBL producing-isolates were resistant to ciprofloxacin and 84.4% to norfloxacin. Among the 96 strains, 80 (83.33%) were found harboring at least one PMQR gene consisting of 78 (81.3%) aac(6’)-Ib-cr, 61 (63.5%) qnrB and 15 (15.6%) qnrS. Among the PMQR-positive strains, 68.4% coharbored qnrB+acc(6’)-Ib-cr genes, 10.5% qnrB+qnrS+acc(6’)-Ib-cr and 6.6% qnrS+acc(6’)-Ib-cr. The qnrB gene was always linked to aac(6’)-Ib-cr gene. Aac(6’)-Ib-cr gene showed the highest association with three ESBL genes (87.6%), followed by qnrB gene (70.6%), then qnrS (17.7%).

Conclusion: The PMQR genes were highly prevalent in ESBL-producing Klebsiella pneumoniae, primarily the aac(6’)-Ib-cr gene. The high associated was observed between ESBL and PMQR genes, notably with the aac(6’)-Ib-cr gene.