Open Access Original Research Article

Metagenomic Study and Biodegrading Capability of Bacterial Community in Monocrotophos Treated Tea Soil

Sangeeta Borchetia, Madhurjya Gogoi, Raktim Pal, Pritom Chowdhury, Afruza Zaman, Hemanta Saikia, Tanoy Bandyopadhyay, Anoop Kumar Barooah

Journal of Advances in Microbiology, Page 1-14
DOI: 10.9734/JAMB/2018/42465

Aims: The study was undertaken to reveal the diversity of bacteria in organophosphate (monocrotophos) pesticide-treated tea soil to provide new insights on monocrotophos degrading bacterial community.

Study Design: A metagenomic study of monocrotophos treated and untreated soil to isolate and identify pesticide-degrading microflora.

Place and Duration of Study: Tea soil was collected from Borbhetta Tea Estate of Tocklai Tea Research Institute, Jorhat, Assam and the experiments were carried out from December 2015 to December 2017.

Methodology: Tea soil was enriched up to 300 ppm of monocrotophos for four weeks (TREATED sample) and 16S rRNA V3 region gene amplicon metagenomic sequencing was carried out on untreated (CONTROL) and spiked (TREATED) soil. The treated soil was cultured in mineral salt medium up to 600 ppm of monocrotophos and bacterial growth, and degrading capacity was studied for three isolated bacterial species at three different pH and identified by sequencing the 16s rRNA region. The bacterial species in the metagenome were also compared and grouped with the bacterial species in NCBI database based on the presence and absence of organophosphate hydrolase (OPH) gene.

Results: Metagenomic sequencing revealed the presence of 20 bacterial phyla distributed across 119 families and 433 genera. 147559 sequences remained taxonomically unclassified suggesting the presence of unique undescribed bacteria. Pesticide-contaminated tea soil was mostly dominant with Acidobacteria, Actinobacteria, Firmicutes, Proteobacteria, Bacteroidetes, Nitrospirae and Verrucomicrobia phyla. The number of species observed in the control and treated soil was 1036 and 910 respectively. Three species were isolated and characterised from the TREATED soil in mineral salt medium with pH 5 to 7 and their monocrotophos degradation was determined by UV-Vis microplate reader and HPLC at 2-time points.

Conclusion: Organophosphate-degrading bacteria namely, Pseudomonas pseudoalcaligenes, Pseudomonas fluorescence, Pseudomonas putida, Serratia species, Cupriavidis metallidurans, Burkholderia species, Achromobacter xylosoxidans, Achromobacter species, Sphingomonas species, Ochrobactrum gallinifaecis, and Brucella species were present in increased numbers in the treated sample. 52.2% monocrotophos degradation was observed in Serratia fonticola in 48 hrs in acidic pH of 5.

Open Access Original Research Article

Studies on Bio-color Production by Pseudomonas aeruginosa Isolated from Soil

M. U. Hizbullah, A. A. Farouq, A. S. Baki, M. U. Dabai, A. Nafi’u, M. K. Nata’ala, G. Mustapha

Journal of Advances in Microbiology, Page 1-12
DOI: 10.9734/JAMB/2018/43068

This study aimed at producing biocolor from soil inhabiting bacteria. Soil samples were screened for isolation of green pigment-producing bacteria and identified molecularly using the standard method. The effect of medium, pH, temperature, incubation time, shaking and static conditions on color production were determined on the isolate, and the pigment was extracted by using chloroform. It was observed that green pigment was produced by Pseudomonas aeruginosa in Nutrient broth at pH 7, after 72 hours incubation and at the temperature of 37°C under shaking condition at 4,000 rpm for 15 minutes. The pigments were characterized and identified as pyocyanin using Thin Layer Chromotography (TLC), Fourier Transformed Infrared Spectroscopy (FTIR) and UV-Visible spectroscopy (UV). The stability of the pigment was tested based on pH and temperature. It was found that the green pigment showed stability at 160°C, 200°C and pH 13.

Open Access Original Research Article

Aerobic Bacteria and Fungi on the Surfaces of a Tertiary Assistance Hospital from Northern Brazil

Layane Gabrielle Coelho de Lima, Michele Alves Sanches, Ana Cláudia Alves Cortez, João Vicente Braga de Souza

Journal of Advances in Microbiology, Page 1-7
DOI: 10.9734/JAMB/2018/42991

Background and Objectives: It is very common that hospital environment is colonised by microorganisms. This colonisation is a potential threat for hospitalised patients, especially in high-risk services. Quantification of microorganisms from surfaces is an important strategy to control the hospital infection.

Materials and Methods: Total 70 samples were collected and seeded in the Mueller Hinton and Sabouraud Agar culture media. Media cultures in Petri dishes were incubated and microbial growth was quantified (CFU/cm2). Bacteria cultures were characterised by Gram stain. Fungal cultures were submitted for micromorphological evaluation and CHROMagar™ incubation.

Results: Bacterial population higher than 250 CFU/cm2 was found in 11 out of 70 samples. Fungi were found in 3 out of 70 samples. Common manipulation surfaces as water drinking unit, shelf and water tap presented the highest contamination rates. Bacterial cultures (n= 95) showed mostly the presence of Gram-negative bacteria (81%). In addition, regarding the fungal cultures (n= 129) mostly filamentous fungi (72%), from Aspergillus genera were obtained.

Conclusion: Most of the investigated hospital surfaces presented low contamination (< 5 CFU/cm2). However, microbiological studies should be regular in critical areas in order to reinforce measures to control and prevent hospital infection.

Open Access Original Research Article

Levels of Biofilm Expression in Klebsiella pneumoniae Isolates Exposed to Herbal Drugs

Monsi, Tombari Pius, Abbey, Samuel Douglas, Wachukwu, Confidence Kinikanwo, Wokem, Gloria Ngozika

Journal of Advances in Microbiology, Page 1-7
DOI: 10.9734/JAMB/2018/42685

Background: There is a continuous rise in antimicrobial resistance globally. Factors responsible for this occurrence especially in developing countries are yet to be properly elucidated. Due to the financial implications of antimicrobials individuals in developing countries such as Nigeria resort to the consumption of herbal drugs to treat infections.

Aims: To investigate the levels of biofilm expressed in Klebsiella pneumoniae isolates pre-treated with herbal drugs.

Methodology: Biofilm assay was performed using 24-well polystyrene microtitre plates which mimic the surface for bacterial attachment. Control and clinical isolates of K. pneumoniae were pre-exposed to different concentrations of herbal solutions (Beta cleanser [Bet], Goko alcoholic bitters [Gab], Goko bitters [Gob], Danko solution [Dan], and Ruzu bitters [Ruz]) (100, 50, 25, 12.5, and 6.25%) in 24-well plate and incubated overnight at 37oC. Cell-to-cell surface attachment of K. pneumoniae was recorded by obtaining a photograph of the inoculum in the 24-well plate. Crystal violet method was used to quantify the levels of biofilm attached to the surface of the 24-well plate. Results were anaylsed using Graph pad prism 5.

Results: Cell-to-cell biofilm formation was seen in different drugs used but higher in Bet and Gob. Bet (25%) and Ruz (50%) showed a significant level of attached biofilm formed compared to untreated control. This results show that Bet, Gob and Ruz has the ability to induce biofilm in K. pneumoniae isolates

Conclusion: Bet, Gob and Ruz could predispose K. pneumoniae to enhance its production of biofilm.

Open Access Original Research Article

Antimicrobial Activity of Ethanolic and Methanolic Extracts of Borassus aethiopium Initial Shoot on Multi-drug Resistant Bacteria and Dermatophytes

Chuku Aleruchi, Maichibi Mugla Salma, Obande Attah Godwin

Journal of Advances in Microbiology, Page 1-7
DOI: 10.9734/JAMB/2018/43199

Aim: To examine the phytochemical properties and chemical profile of the methanol and ethanol extracts of the initial shoot of Borassus aethiopium and to evaluate its antimicrobial activity on selected multidrug-resistant pathogenic bacteria and dermatophytes.

Place and Duration of study: Sampling was done from Plateau State, Nigeria, while the analysis was done at the Federal College of Forestry and National Veterinary Research Institute Vom, Plateau State Nigeria between August and December 2017.

Methodology: The chemical profile of B. aethiopium was determined using Gas Chromatography-Mass Spectrometry (GC-MS). Phytochemical analysis was performed to confirm the bioactive components present while the antimicrobial activity was determined using agar well diffusion method using Staphylococcus aureus, Pseudomonas aeruginosa, Klebsiella pneumoniae, Proteus mirabilis, Trichophyton mentagrophytes and Epidermophyton floccosum as the test organisms. Minimum Inhibitory Concentration (MIC) was determined.

Results: All test organisms were susceptible to both the extracts of B. aethiopium. Klebsiella pneumoniae was found to be most susceptible (33 mm) in the methanolic extract and Staphylococcus aureus as the most susceptible (31mm) in the ethanolic extract while Epidermophyton floccosum was the least susceptible of both the extracts (19mm; 20mm). Methanolic extract of B. aethiopium showed the lowest MIC on the test organisms compared to the ethanolic extract. The difference in the chemical composition of the extracts obtained was Tridecanoic acid, Valeric acid, 14-Octadecenoic acid, Heneicosanic acid, Erucic acid, and Oxacyclotetradecane for the methanol extract and Dodecanoic acid, Undecanoic acid, Docosanoic acid, Oleic acid, 9-Octadecenoic acid (Z)- and 9,12,15-Octadecatrienoic acid for the ethanol extract. Bioactive components present in B. aethiopium extracts (%) were saponins 14.40; 6.45, flavonoids 24.20; 12.40, cardiac glycosides 13.55; 8.20, steroids 6.11; 4.12, alkaloids 5.00; 2.00, terpenoid 12.50; 11.80 and tannins 1.20; 0.05.

Conclusion: The initial shoot of B. aethiopium has the potential as a source of antimicrobial essential to the pharmaceutical industries. However, the toxicological analysis is recommended to assess the toxicity and safety on sensitive organs of the animals.