Open Access Original Research Article

Antimicrobial, Phytochemical Analysis and Molecular Docking (In-silico Approach) of Tithonia diversifolia (Hemsl.) A. Gray and Jatropha gossypiifolia L on Selected Clinical and Multi-Drug Resistant Isolates

A. F. Okiti, O. T. Osuntokun

Journal of Advances in Microbiology, Page 1-18
DOI: 10.9734/jamb/2020/v20i630248

The aim of this study is to determine the zones of inhibition, phytochemical screening and molecular docking (In-silico Approach) of Tithonia diversifolia (Hemsl.) A. Gray and Jatropha gossypiifolia L against selected clinical and multi drug resistant isolates. Crude extraction of air dried leaves were carried out by soaking the plant in ethanol and ethyl acetate, standard agar diffusion method was used for sensitivity testing, minimum inhibitory concentration and minimum bactericidal concentration values were obtained by agar dilution method. The antimicrobial activity of the leaf extracts of T. diversifolia (Hemsl.) A. Gray and J. gossypiifolia L was assayed against Bacillus subtilis, Escherichia coli, Klebsiella pneumonia, Proteus mirabilis, Pseudomonas aeruginosa, Salmonella typhi, Shigella dysentriae, Staphylococcus aureus, Streptococcus pyogenes, Candida albican and against multi drug resistant bacteria which are Acinetobacter baumannii, Enterobacter agglomerans, Proteus mirabilis, Providencia stuartii, Salmonella subsp 3b. Levofloxacin and fluconazole were the standard antibiotics used. Sensitivity test revealed the highest zone of inhibition observed for J. gossypiifolia L and T. diversifolia (Hemsl.) A. Gray against Candida albican with mean and standard deviation of 29±1.414 and 19.5±0.707 at 100 mg/ml respectively, while the least zone of inhibition was observed from the extracts of J. gossypiifolia L against Escherichia coli with 11.75±0.354 at 100 mg/ml. Both plant extracts showed antimicrobial activity against multi drug resistant isolates having zones of inhibition ranging from 0 to 15±1.414. The Minimum Inhibitory Concentration of the extracts ranges between 6.25 and 100 mg/ml as well as the Minimum Bactericidal Concentration. The qualitative and quantitative phytochemical analysis showed the presence of alkaloids, anthraquinone, cardiac glycosides, flavonoids, phlobotannins, reducing sugars saponins, steroids and tannins. Molecular docking of the phytochemicals of T. diversifolia (Hemsl.) A. Gray only was carried out using levofloxacin as template, which revealed the presence of compounds more effective in inhibiting DNA gyrase enzyme. Thus, the use of both plants as traditional medicine is justifiable and should be encouraged in the formulation and production of new antibiotics.

Open Access Original Research Article

Early Onset and Late Onset of Neonatal Sepsis in a Tertiary Hospital, South-South, Nigeria

A. A. Adedokun, E. O. Onosakponome, C. U. Nyenke

Journal of Advances in Microbiology, Page 19-29
DOI: 10.9734/jamb/2020/v20i630249

Aims: The study was carried out to determine organisms present during early onset of neonatal sepsis (EONNS), late onset of neonatal sepsis (LONNS) and their antimicrobial susceptibility pattern.

Methodology: This study is a retrospective evaluation of 453 neonatal blood cultures. Two (2) ml of blood from these neonates was cultured in thioglycollate broth and tryptone soya broth. This was carried out in the Department of Microbiology and Parasitology of the University of Port Harcourt Teaching Hospital, Nigeria between January-December 2007.

Results: Out of 453, 272(60.0%) neonates showed negative blood cultures, while 181(40.0%) neonates had positive bacterial cultures and 2 neonates (0.4%) were positive for Candida albicans. Overall, Klebsiella spp. was the most common pathogen, accounting for (37.8%) of the total isolates. Others were as follows; Staphylococcus aureus (28.4%), Escherichia coli (11.8%), unclassified coliforms 8.3%, Pseudomonas spp. 4.9%, Enterococcus spp. (2.9%), coagulase-negative Staphylococcus (CONS) (2.5%) and Proteus spp. (3.4%). Early onset neonatal sepsis (EONNS) and late onset neonatal sepsis (LONNS) had Klebsiella spp and Staphylococcus aureus as their common causes of neonatal sepsis respectively. Klebsiella spp. was susceptible to spafloxacin (87.0%) followed by ofloxacin (82.0%), amoxycillin-clavulanic acid (79.0%) and ceftazidime (65.0%) among the Gram-negative organisms. In the category of Gram-positive organisms, Staphylococcus aureus is highly sensitive to ofloxacin (81.0%) followed by spafloxacin (79.0%) and amoxicillin-clavulanic acid (71.0%).

Conclusion: A viable antibiotic susceptibility surveillance programme coupled with good infection control practices and rational antibiotics use will reduce infection rate, ensure better therapeutic success and prolong the efficacy of available antimicrobials.

Open Access Original Research Article

Studies on the Proximate Contents and Potentials of Musa sapientum and Persea americana Fruits Peel as Media for the Growth of Medically Important Fungi

David C. Ekwe, Osmond C. Ekwebelem, Emmanuel A. Eze

Journal of Advances in Microbiology, Page 30-36
DOI: 10.9734/jamb/2020/v20i630251

Aim: The present study was designed to formulate growth media for medically important fungi using banana and avocado fruits peel.

Methodology: The peels were obtained from fresh banana and avocado fruits and dried into crisps at 100oC using hot air oven, pulverised into powder using Hammer mill machine and sieved into fine powder using 1 mm sieve size. Pure isolates of Aspergillus flavus, Aspergillus fumigatus and Cladosporium sp. were obtained from the Diagnostic Laboratory of the Department of Microbiology, University of Nigeria, Nsukka and used for the study. Two different media, Banana Peel Agar (BPA) and Avocado Peel Agar (APA) were formulated. 2 g of each fruit peel powder was introduced into separate conical flasks, 1g of agar powder was added as solidifying agent and varied glucose concentrations (20%, 10%, 5% and 0%) was used as nutrient (carbon source) supplement. The prepared media was sterilized by autoclaving at 121°C for 15 minutes. The test organisms were inoculated aseptically onto the formulated media and incubated for 3 days at 37oC. Fungal growth was visually observed. For comparative analysis, Sabouraud dextrose agar (SDA) was used as control. 

Results: The formulated media supported the growth of all the test organisms, although at varying degrees. BPA showed more suitability than APA, with the growth of the test organisms on BPA favourably comparable to that on SDA. Proximate analysis of the fruit peels indicated that the moisture, ash, fat, crude fibre, crude protein and carbohydrate contents of avocado peel were 7.35%, 14.65%, 0.10%, 9.75%, 6.57% and 61.58% respectively and 5.80%, 6.00%, 0.45%, 5.35%, 10.95% and 71.45% respectively for banana peel.

Conclusion: Banana and avocado fruits peel can be utilized as alternative materials in the formulation of culture media for the in vitro cultivation of fungi for medical and research purposes.

Open Access Original Research Article

Molecular Characterization and Detection of Antibiotic Resistance Genes in Pseudomonas Species Isolated from Tympanotonus fuscatus

T. Sampson, N. P. Akani, I. O. Hakam

Journal of Advances in Microbiology, Page 37-45
DOI: 10.9734/jamb/2020/v20i630252

Aim: This was carried out to characterize Pseudomonas species isolated from the West African Mud Creeper (Tympanotonus fuscatus) molecularly and as well detect the possible presence of inducible AmpC gene that mediates resistance to cephalosporins and most penicillins.

Sample: Tympanotonus fuscatus (West African Mud Creeper), a gastropod mollusc found in brackish waters of West Africa was used for the study.

Place and Duration of Study: This study was carried out between February and August 2019 at the Department of Microbiology, Rivers State University, Port Harcourt, Nigeria.

Methodology: Thirty two (32) Pseudomonas species were isolated and identified culturally from T. fuscatus. Pseudomonas species isolates were subjected to a group of ten (10) antibiotics using the Kirby-Bauer disc diffusion method and resistant isolates were screened molecularly for the presence of resistance gene (AmpC). AmpC screening was carried out in a step wise process of DNA extraction, quantification, amplification of ampC gene using appropriate primer and Agarose gel electrophoresis to reveal which DNA extracts had ampC genes amplified. The two most resistant isolates had their 16S rRNA sequenced, identified and were also profiled for plasmids by extracting plasmid DNA.

Results: Results revealed that 96.67% of isolates had MAR index greater than 0.2 indicating high a risk source of contamination where antibiotics are often used. Results also showed the presence of ampC gene in seven (7) out of the twelve (12) isolates screened for ampC gene. Molecular characterization via sequencing of the 16S rRNA gene of the two (2) most resistant isolates confirmed that both isolates were strains of Pseudomonas aeruginosa. Profiling of plasmids also revealed the presence of plasmid DNA of about 10 kilo base pairs in both isolates profiled.

Conclusion: This study has revealed the resistance ability of Pseudomonas and some reasons behind this resistance. Appropriate investigation into antimicrobial resistance is recommended for the administration of drugs for the treatment of food-mediated Pseudomonas infections.

Open Access Original Research Article

Production, Optimization, Purification and Characterization of Mannanase Produced by Bacillus Subtilis

Adebayo-Tayo Bukola Christianah, Onifade Deborah Ajoke

Journal of Advances in Microbiology, Page 52-67
DOI: 10.9734/jamb/2020/v20i630254

Endo-β-1,4-D-mannanase (β-mannanase; EC catalyses the random hydrolysis of mannoglycosidic bonds in mannan-based polysaccharides. These enyzmes are commonly found in nature and are located within the structure of mannans and heteromannans (galactomannan, glucomannan and galactoglucomannan) in the hemicellulose fraction of trees with soft tissues and hard tissues, locust bean seed.  Most β-mannanases degrade mannooligosaccharides down to a degree of polymerization of four. In this study mannanase was produced using a submerged fermentation method from Bacillus subtilis. The effect of growth of the organism and processing parameters on the production of mannanase were determined after which optimization studies were carried out. The enzyme was partially purified using ammonium sulphate, dialysis and gel filtration. The partially purified enzyme was characterized. The result of the study showed that mannanase enzyme from Bacillus subtilis was optimally produced in a medium comprising of galactose, peptone, sugarcane bagasse at a pH of 6.0 and a temperature of 45 oC. The enzyme was most stable and active at pH 10.0 and at a temperature of 40 oC.  and 60 respectively.

Open Access Original Research Article

Detection of Multidrug Resistant Gram Negative Bacteria in Healthy Cattle from Maiduguri Metropolitan, Nigeria

Adam Mustapha, Mustafa Alhaji Isa, Ibrahim Yusuf Ngoshe, Hashidu Bala

Journal of Advances in Microbiology, Page 68-76
DOI: 10.9734/jamb/2020/v20i630255

Aim: Prevalence of multidrug resistant bacteria on apparently health animals has turned antibiotic resistance to multifaceted process and threatens global food security and public health. The aim of the present study was to investigate the resistance profile of isolates from apparently healthy cattle in Maiduguri, Nigeria.

Methodology: A total of 120 nasal swab samples were collected from cattle. Colony identification was according to the guidelines of Bergey’s Manual of Determinative Bacteriology. The susceptibility pattern of the isolates was conducted on the identified isolates according to the Modified Kirby-Baur disc diffusion method on Muller-Hilton agar and interpreted according to the procedures of Clinical Laboratory Standards Institute (CLSI, 2018) guidelines. Multiple Antibiotic Resistance Index (MARI) was calculated using the formula, MARI=a/b where “a” is the number of antibiotic resisted and “b” is the total number of antibiotic used in the study.

Results: Of the total samples (120) from cattle 96 (80%) detected the following isolates; E. coli was the most commonly recovered isolates (33, 34.4%), followed by Klebsiella spp (28, 29.2%), Salmonella spp (21, 21.9%) and Pseudomonas aeruginosa (14, 14.5%). In this study, all the recovered isolates were found to be multidrug resistant gram negative bacteria, with highest resistance was shown by Salmonella spp. The high MARI observed in all the isolates in this study ranging from 0.7 to 0.9. MARI value of 0.2 > is suggests multiple antibiotic resistant bacteria and indicate presence of highly resistant bacteria.

Conclusion: The study indicates highly resistant bacteria are carried by healthy food animals. Thus, there is need for continued monitoring of antibiotics use in animal husbandry to prevent further spread of resistance in Maiduguri, Nigeria.

Open Access Original Research Article

Bacteria Associated with Petroleum Hydrocarbon Contaminated Soil from NNPC Depot

S. O. Bankole, M. B. Oyedeji, F. O. Akanni, E. P. Chukwudebe, A. O. Olatunji, O. S. Ariwoola

Journal of Advances in Microbiology, Page 77-82
DOI: 10.9734/jamb/2020/v20i630257

The microorganisms associated with soil polluted with petroleum hydrocarbon were isolated at NNPC depot Ibadan, in Ido Local Government Areas of Oyo State. Soil samples were taken from three different points along the point of discharge. The spread plate method was used to isolate the microorganisms found in these rivers and were later identified. From the research, seven different species of microorganism were isolated, which are Pseudomonas sp, Bacillus sp., Micrococcus sp. Staphylococcus aureaus sp., Enterobacter sp., Klebsiella sp. and Escherichia coli. Pseudomonas species were found to be more prominent in the samples collected in a research work. This suggested that the isolates are resistant to the petroleum hydrocarbon and can be recommended as candidates for the clean-up of petroleum hydrocarbon contaminated soil.

Open Access Original Research Article

Re-thinking Metronidazole: Anti-microbial Synergism of Hexane Extracts of Garcinia kola and Aframomum melegueta

Joy I. Odimegwu, Babatunde Asabisi, Mariam Sodeinde, Margaret O. Ilomuanya

Journal of Advances in Microbiology, Page 83-93
DOI: 10.9734/jamb/2020/v20i630266

Aims: This research evaluates knowledge, attitudes, and practice with regard to food poisoning treatment. It compares the antimicrobial effects of Garcinia kola (Gk) and Aframomum melegueta (Am) seeds with Metronidazole and Gentamicin as standards.

Study Design: A survey with questionnaires was carried out among community Pharmacists in Lagos, Nigeria on drug of choice for suspected food poisoning and the responses analyzed. Hexane extracts of Gk and Am seeds were used on selected microbials that cause food poisoning.

Place and Duration of Study: Department of Pharmacognosy; Phytochemistry Laboratory and Department of Pharmaceutics and Pharmaceutical Technology; Pharmaceutical Microbiology Laboratory, Faculty of Pharmacy. University of Lagos.

Methodology: Agar diffusion assays were carried out using and using Gk and Am with Metronidazole and Gentamicin as controls. Gas chromatography and Mass spectroscopy of the extracts was also carried out to ascertain the possible active phytocomponents.

Results: Our discovery on knowledge showed that 98.6% of the interviewed Pharmacists use Metronidazole for food poisoning treatment. 41.7% of the respondents prescribe Ciprofloxacin while 66.7% prescribed Tetracycline as adjunct medications. Gk and Am exerted powerful anti-microbial effects in a dose dependent manner. Gk on S. typhi showed 1.4-1.7 cm zone of inhibition (zi) at 200mg/ml and with E. coli, 1.2 cm at same concentration. Am with S. typhi showed1.2-1.8 cm and E. coli 1.5 cm zi at same concentration too, much better than Gk. Their mixture for the synergistic experiments had 2.0-2.2 cm and E. coli, 2.4 cm zi at 200 mg/ml, the best performance of the extracts as anti-infective agents against food poisoning. 19 compounds were discovered through GC/MS in Gk and 31 in Am. They each exerted more antimicrobial effects than Metronidazole and their mixture used for synergistic experiments compared favorably with Gentamicin.

Conclusion: Hexane extracts of Gk and Am are good alternatives for the treatment of food poisoning. The study encourages further drug discovery research in order to ascertain the particular bioactive compounds in the extracts responsible for their powerful antimicrobial actions. Metronidazole is not the drug of choice for food poisoning management.

Open Access Review Article

Coronavirus: COVID-19-Epidemiology, Treatment, Prevention and Control

Udegbe, Chinemerem Dorathy, Effiong, E. Bassey

Journal of Advances in Microbiology, Page 46-51
DOI: 10.9734/jamb/2020/v20i630253

Coronavirus infection is a zoonotic disease of viral origin. The SARS-CoV-2 is the novel virus that causes the ongoing pandemic affecting the global community hence a very big threat to the global public health. Coronavirus disease 2019 (COVID-19) was first detected Wuhan, China and reported to World Health Organization (WHO) country office in China on 31st December 2019. It has since been spreading, resulting in an ongoing pandemic. As at 19 May, there are about 4.8 million cases of Covid-19 with 316,169 deaths reported in more 188 countries of the world. The initial transmission appeared to be from an animal source, but there has been person to person transmission in the affected countries. A lot of preventive and control measures have employed to severe transmission. Strategies in the control of an outbreak are containment or suppression and mitigation. Currently there is no specific vaccine or treatment for COVID-19. This work reviewed the Coronavirus Disease 19, its epidemiology, pathogenesis, symptoms, diagnosis, treatment, prevention and control.