Open Access Original Research Article

Natural Resistance Associated Macrophage Protein-1 Expression in Salmonella typhi Infection with Acute Recurrence State of Typhoid Fever in Endemic Areas, Indonesia

Ade Rifka Junita, Firdaus Hamid, Rosdiana Natzir, Rosana Agus, Burhanuddin Bahar, Mochammad Hatta

Microbiology Research Journal International, Page 1-10
DOI: 10.9734/mrji/2020/v30i930258

Background and Aim: The molecular pathogenesis of typhoid fever is complex and the host susceptibility mechanisms such as Natural Resistance Associated Macrophages Protein-1 (NRAMP-1) are poorly understood. This study explores the expression of NRAMP-1 in the serum of Acute Recurrence State (ARS) of typhoid fever compared with typhoid fever patients and healthy persons.

Methods: Thirty of ARS of typhoid fever and 30 typhoid fever patients were collect from several Primary Health Care and Hospitals in the endemic area. Diagnosis of typhoid fever based on clinical symptoms and confirmed by blood culture. Healthy persons were collect from the Blood Transfusion Unit, Makassar, Indonesia. The expression of NRAMP-1 was determined using Enzyme-linked immunosorbent assay (ELISA).

Results: The mean of NRAMP-1 expression in 30 ARS of typhoid fever and 30 typhoid fever were found 10.941,56 pg/mL and 11.027,65 pg/mL, respectively. However, the mean of NRAMP-1 expression in healthy persons was found 21.103,91 pg/mL.

Conclusion: No different the expression of NRAMP-1 in ARS of typhoid fever compared with typhoid fever patients. However, expression of NRAMP-1 in both ARS of typhoid fever and typhoid fever showed significantly lower than healthy persons. Future study is needed to explore the other molecular factors involved to become ARS.

Open Access Original Research Article

Multiple-Antibiotic Resistance and Presence of CTX-M Genes among Enterobacteriaceae Isolates from Different Sources in Iwo, Osun State, Nigeria

Adeleke, Olufeyikemi A., Owoseni, Abimbola A.

Microbiology Research Journal International, Page 30-38
DOI: 10.9734/mrji/2020/v30i930260

Aims: Assessment of the antibiotic resistance pattern and detection of genes responsible for the β-lactam resistance in enterobacteria isolated from different sources was carried out in the course of the study.

Place and Duration of Study: Samples from different sources in Iwo, Osun State, Nigeria including Abattoir and Cow ranch. The study was done August 2015 to May 2016.

Methodology: Isolation was done on nutrient agar, xylose lysine deoxycholate (XLD) and identified using standard procedures. Antibiotic multidisc containing the following ceftazidime (30 µg), cefruroxime (30 µg), gentamicin (10 µg), ciprofloxacin (5 µg), ofloxacin (5 µg), augmentin (30 µg), nitrofurantoin (300 µg), and ampicillin ( ) were used to study the resistance patterns of the bacteria. Polymerase chain reaction (PCR) method was carried out to screen for CTX-M gene in beta-lactam resistant isolates and statistical analyses were carried out using the analysis of variance test (ANOVA) and the PAST (paleontology statistics) software analytical package where applicable.

Results: Fifty-two (52) entrobacteria consisting of nine genera (9) were isolated and identified, Serratia (13), Enterobacter (11), Klebsiella (6), E. coli (5), Proteus (5), Salmonella (3), Shigella (3), Citrobacter (3), and Yersinia (3). The highest resistance observed by the isolates was to ampicillin (80.7%) followed by augmentin (40%) and 33% resistance of the isolates to cefuroxime, while all the isolates were sensitive to ceftazidime, gentamicin, ofloxacin, ciprofloxacin and nitrofurantoin. Nineteen of the isolates were resistant to three or more of the tested antibiotics i.e. multiple resistance while five of the multidrug resistant isolates show the presence of CTX-M β-lactam resistance gene which seems to be responsible for their resistance to the β-lactam antibiotic tested.

Conclusion: There are a number of organisms in the environment that have developed multiple resistances to several antibiotics and in view of this thwart treatment; of major concern are the Enterobacteriaceae family in which new resistance characteristics have been discovered leading to increasing resistance to standard available drug therapies.

Open Access Original Research Article

Microbiological Research of the Effects of Electromagnetic Fields of the Bacteria Escherichia coli, Enterococci, Coliforms and Clostridium perfringens

Nedyalka Valcheva, Ignat Ignatov

Microbiology Research Journal International, Page 39-44
DOI: 10.9734/mrji/2020/v30i930261

The aim of the study was to analyze the microbiological effects of electromagnetic fields with method of Drossinakis with reliable microbiological methods. A comparative analysis of the number of bacteria of a given species before and after electromagnetic influence was performed. For the research was applied water with the following bacteria –Escherichia coli, Enterococci, Coliforms and Clostridium perfringens.

The method of Drossinakis is including the electromagnetic waves in the electromagnetic spectrum, from 1 Hz to 3.1015 Hz. For the inhibiting effects on bacteria the method of Drossinakis is including low frequencies from 20 till 70 Hz.

The study shows the effects with low frequencies electromagnetic waves for decreasing of the number of bacteria. The controlled microbiological parameters are defined by the membrane method and by using of differential diagnostic nutrient media at 24 hours.

Open Access Original Research Article

Evaluation of the Symbiotic Efficiency of Rhizobium Strains Applied to Bituminaria bituminosa (L.) Stirton

Omar Zennouhi, Abderrazak Rfaki, Mohamed El Mderssa, Jamal Ibijbijen, Laila Nassiri

Microbiology Research Journal International, Page 45-55
DOI: 10.9734/mrji/2020/v30i930262

Aims: The study aimed to evaluate the effect of inoculation by different rhizobacteria on Bituminaria bituminosa plants grown under greenhouse conditions.

Study Design: An experimental study.

Place and Duration of Study: The study was carried out at the Department of Biology (Environment and valorization of microbial and plant resources Unit), Faculty of Sciences, Moulay Ismail University-Meknes, from November 2019 to February 2020.

Methodology: Eleven species and/or isolates belonging to Rhizobium genus are used to inoculate B. bituminosa plants; similarly, fresh and dry crushed nodules previously collected from B. bituminosa shrubs are tested. The bacterial inoculation effects are evaluated through the estimation of inoculated plants’ fresh and dry shoots weight, root dry weight, total nitrogen, nodules number and fresh weight in comparison to non-inoculated plants. The infectivity and efficiency of the bacteria and the biological nitrogen fixation are also evaluated.

Results: The results enable us to select the infective strains on the basis of their positive effect on growth and total nitrogen, in order to produce inoculum for B. bituminosa. Efficiency and biological nitrogen fixation are also very high compared to the control, especially with the B.b1 strain isolated from Bituminaria bituminosa and identified as Rhizobium tibeticum. The fresh nodules crushing is also very efficient. as inoculant.

Conclusion: The use of symbiotic complex as Rhizobium tibeticum – Bituminaria bituminosa or an inoculum produced from fresh nodules are an eco-friendly alternative for the design of sylvo-pastoral systems ensuring increased soil fertility, fodder productivity and sustainable agroforestry.

Open Access Original Research Article

In vitro Antifungal Activity of Combination of Miconazole, Salicylic Acid and Benzoic Acid in Two Different Bases against Trichophyton mentagrophytes

Hassan Thoulfikar A. Alamir, M. E. L. A. Shayoub, Wisal G. Abdalla, Eltayeb Suliman Elamin

Microbiology Research Journal International, Page 56-61
DOI: 10.9734/mrji/2020/v30i930263

In vitro activity of miconazole, salicylic acid and benzoic acid against T.richophyton mentagrophytes was done on Sabouraud’s dextrose agar. The inhibition zones were measured in cm. 2.8 cm, 1.5 cm and no inhibition zone were reported for miconazole, salicylic acid and benzoic acid respectively. Synergistic effect of salicylic acid and benzoic acid against T. mentagrophytes was done using two different bases of cream lanette 20% and aqueous cream 30/70. Four different formula each one contain 2% miconazole with different concentrations of salicylic acid and benzoic acid were used. Biggest inhibition zone (5 cm) was observed of lanette cream which contains 2% miconazole, 3% salicylic acid and 6% benzoic acid.

Open Access Original Research Article

Expression of Epsilon Toxin from Clostridium perfringens Type D in E. coli Rosetta (DE3): Study on a New Vaccine Production

Maryam Adibi, Reza Pilehchian Langroudi, Lida Abdolmohammadi Khiav

Microbiology Research Journal International, Page 62-69
DOI: 10.9734/mrji/2020/v30i930264

Clostridium perfringens is a Gram-positive anaerobic bacterium which is divided based on major toxins into five types (A-E). C. perfringens type D causes fatal enterotoxaemia (pulpy kidney) in sheep and goats that causes heavy economic losses in domestic animals. However current enterotoxaemia vaccine has been effective in controlling of disease; Strategies for the development of effective vaccine can be achieved by the production of new generation experimental vaccines. The aim of this study was cloning and expression of epsilon toxin of C. perfringens type D in E. coli Rosetta (DE3) to improve of the immunity. Epsilon toxin gene was cloned into pJET1.2/blunt vector and pET22b (+) expression vector and finally transformed into E. coli Rosetta competent cells (DE3). The result showed that the epsilon gene of C. perfringens type D can be cloned and expressed in E. coli RosettaTM (DE3) successfully. E. coli was suitable host for the expression of C. perfringens epsilon toxin. Clearly, future research is open for further refinement of the study.

Open Access Original Research Article

Bacteriological Profile of Nosocomial Infections in Visceral Surgery at the CNHU-HKM of Cotonou in Republic of Benin

Aïkou Nicolas, A. Ahoyo Theodora, Degbe Sah Cyriaque, Coulibaly Founzégué Amadou, Sezan Alphonse, Edorh A. Patrick, R. A. Lapo

Microbiology Research Journal International, Page 70-77
DOI: 10.9734/mrji/2020/v30i930265

Wound suppuration is the formation and accumulation of pus in the soft tissue of the wound. As a rule, the natural inflammatory reaction disappears within three to five days and later the wound heals without any character. During this time, in some cases the inflammatory reaction in the wound exceeds the limits of the physiological norm due to the external supply of bacterial germs, which leads to the formation of pus, exaggeratedly prolonging the healing process and in turn leading to serious consequences. This study aims to identify the bacteria involved in these suppurations, to study their antibiotic resistance profiles and to review the role of the nurses taking care of these patients. The study was conducted at the Menontin zone hospital from May 15 to June 21, 2013. Fifty-one patients of all ages and sexes with surgical and traumatic discharge wounds were sampled and 30 hand samples were taken from the nurses' hands before and after dressing. From the results obtained, 76 bacteria were identified, including 35 cocci and 41 bacilli. 29% of the bacteria found were Pseudomonas aeruginosa, 28% Staphylococcus aureus, 14% Escherichia coli and others. Hand samples of 10 bacteria were isolated including 4 Pseudomonas aeruginosa, 3Staphylococcus DNase-negative 2 Klebsiella spp and 1 Staphylococcus aureus. The strains isolated were particularly resistant to the antibiotics tested more specifically to β-lactam, Gentamycin Erythromycin and Colistin. The majority of strains have good sensitivity to ciprofloxacin and Fosfomycin. In view of these results, it is necessary to draw the attention of patients and nurses to the reality of the existence of these bacteria, the adequate intake of antibiotics preferably after an antibiotic test, hand washing and the use of appropriate sterile materials before and after any dressing.

Open Access Original Research Article

Antifungal Activities of Vernonia amygdalina Crude Extracts and Fractions against Strain1161, P37005 and RM1000

A. D. M. Owoyale, M. Galadimma, S. Y. Daniyan, N. Adabara

Microbiology Research Journal International, Page 78-93
DOI: 10.9734/mrji/2020/v30i930267

This research study was focused in determining antifungal activities of the crude extracts and fractions of Vernonia amygdalina against strain 1161, P37005 and RM1000. Vernonia amygdalina leaves were extracted by the reflux extraction protocol which was done in a successive method. This study also analysed quantitatively the phytochemical that were present in the crude leaf extracts of Vernonia amygdalina. The study identified the presence of tannin, flavonoid, alkaloids, saponin and phenols. The three strains used in this study were tested for their susceptibility, However, activity of the crude extract was assayed at a varying concentration of 40, 60, 80 and 100 mg/ml. The methanol leaf extract (MLE) at a concentration of 100mg/ml showed the highest zones of inhibition 15.33± 1.23 mm against strain 1161(isolate S5). The result of the minimum inhibitory concentration (MIC) and minimum fungicidal concentration showed for active crude extracts were 12.5 mg/ml and 50 mg/ml presented for the n- hexane crude extract against strain 1161(IsolateS5). The value for the methanol crude extract were 12.5 mg/ml and 50 mg/ml against strain 1161 (Isolate: S5). Furthermore, the value of 12.5 mg/ml and 100mg/ml were showed for n- hexane crude extract against P37005 (isolate: B4) However, The antifungal activity of the fractions against the strains were determined at a concentrations of 5mg/ml and 10mg/ml. The various fractions of Vernonia amygdalina showed inhibitory activity against all the strains.16.00± 0.0mm was the highest value that was presented for n-hexane fraction of Vernonia amygalina against strain 1161 and P37005. The result of the minimum inhibitory concentration (MIC) and minimum fungicidal concentration (MFC) for active fractions (F1) are 12.5 mg/ml and 25 mg/ml showed for n- hexane fraction of Vernonia amygdalina against strain P37005(Isolate:B4). The value of 12.5 mg/ml and 25 mg/ml were showed for n- hexane fraction (F4) of Vernonia amygdalina against strain 1161 (isolate: S5). Fraction (F4) showed 3.125 mg/ml and 6.25 mg/ml against strain P37005 (isolate: B4). The methanol fraction (F1) showed values of 3.125 mg/ml and 25 mg/ml respectively against strain RM1000(isolate B2). Fraction F6 had values of 3.125 mg/ml and 6.25 mg/ml against strain P37005 (isolate: B4). It can be deduced from this study that the n- hexane and methanol crude extracts showed a significant result than the ethyl acetate crude extracts. Furthermore, there was inhibitory activity for the n-hexane, ethyl acetate and methanol fractions against all the tested strains.

Open Access Original Research Article

Detection of Clostridium difficile Infection in Al-Quwayiyah General Hospital, Riyadh, Kingdom of Saudi Arabia

Enas Sh. Khater, Abd Alazim A. Al- Faki

Microbiology Research Journal International, Page 94-102
DOI: 10.9734/mrji/2020/v30i930268

Clostridium difficile infections (CDIs) is considered healthcare-associated infections which cause watery diarrhea to long stayed hospitalized patients and cause increased mortality rate.

Aim: Detection of the prevalence and risk factors of C. difficile in Al Quwayiyah General hospital, Riyadh, Kingdom of Saudi Arabia and compairing between GeneXpert® PCR assay and Quikchek complete-enzyme imunoassay QCC, (QCC-EIA) in detection of C. difficile infection and toxicity

Materials and Methods: A cross sectional and prospective study was performed for one year started from June 2019 to June 2020. The data collected include demographic, laboratory and clinical data. A total of 104 stool samples were collected from patients presented with diarrhea. GeneXpert® PCR assay and Quikchek complete-enzyme imunoassay QCC (QCC-EIA) were conducted to each stool sample.

Results: Only 15(14.4%) of the 104 studied patients had CDI while 89 (85.6%) were non CDI patients, 13 (86.7%) of the CDI patients were males and 2 (13.3%) were females with mean age for CDI cases 61 (±19.9), while non CDI cases involved 55(61.8%) were males and 34 (38.2%) were females with mean age for cases of non CDI, 60 (±18.7) years. Of the CDI and non CDI cases respectively 12 (80%) and 14(15.7%) had fever, 5 (27%) and 6 (6.7%) had vomitting and 7 (46.7%) and 12 (13.5%) of cases had abdominal pain. There was statistical significant difference between patients with fever while no statistical significant difference regarding vomitting and abdominal pain. There was statistical significant difference between patients with peptic ulcers, patients received proton pump inhibitors and patients received broad-spectrum antibiotics, while There was no statistical significant difference between cardiac disease, cerebrovascular disease, diabetes, pulmonary disease, hepatic disease and Renal disease. Gene expert PCR detected 15/104(14.4%) as positive CDI while QCC-EIA detected 21/104 (20.5%) as positive CDI. On comparison between gene expert PCR technique and QCC-EIA the sensitivity of QCC-EIA was 100%, while the specificity was 91%. The Positive Predictive Value was 74%, while the Negative Predictive Value was 100%.

Conclusion: The C. difficile infection prevalence rate in the hospital was 14.4%. There was statistical significant difference between patients with peptic ulcers, patients received proton pump inhibitors and patients received broad-spectrum antibiotics. The QCC-EIA can be used as a screening test for the detection of C. difficile toxin in stool samples but should be confirmed with a PCR assay or another confirmatory test Due to its decreased specificity.

Open Access Original Research Article

Detection and Prevalence of Canine Leptospirosis from Navsari District of South Gujarat, India

Dhruv Desai, Pushpa Makwana, Jayesh Solanki, Irshadullakhan Kalyani, Dharmesh Patel, Sudhir Mehta, Suresh Mavadiya, Jignesh Vala, Saurabh Parmar

Microbiology Research Journal International, Page 103-110
DOI: 10.9734/mrji/2020/v30i930269

Aims: Leptospirosis is endemically prevalent in coastal area of south Gujarat. Generally, pets are vaccinated against Leptospira interrogans but whosoever left unvaccinated, are prone to infection. The present study was conducted to detect canine leptospirosis and its prevalence from south Gujarat.

Place and Duration of Study: A total of 46 serum samples and 33 urine samples were collected from 56 dogs suspected of having leptospira infection. The study was done from September 2019 to May 2020.

Methodology: Serum samples were diagnosed by IgG based ImmunoComb solid phase dot ELISA test for antibody against genus specific Leptospira interrogans. Urine samples were visualized under dark field microscopy (DFM) following laboratory manual on leptospirosis of WHO. Sex and age of dogs were noted to find out the percent positivity and prevalence. Chi square test was applied to find out any significant difference among the age groups.

Results: Out of 46 serum samples, 17 (36.95%) found positive by dot ELISA and out of 33 urine sample, 13 (39.39%) found positive by DFM. Sex wise prevalence found 42.10 percent (16/38) in Male while 55.55 percent (10/18) in female dogs. Age group wise prevalence was recorded 63.63 percent (07/11) in 1- ≤ 3 years group. 27.77 per cent (05/18) in > 3- ≤ 6 years group whereas, 51.85 percent (14/27) in > 6- ≤ 12 years group.

Conclusion: Detection and prevalence of canine leptospirosis were studied first time from Gujarat. Overall 46.42 percent (26/56) dogs were diagnosed positive for canine leptospirosis. Dogs having age group of 1- ≤ 3 years found more susceptible. Prevalence was found more in female than male dogs. 

Open Access Original Research Article

Isolation, Biochemical Characterization and Production of Immobilised β-Amylase Chitosan Beads Using Bacteria from Waste Water Effluents for its Industrial Production Aspect

Jyoti Chandola, Vandana Semwal, Narotam Sharma, Pooja Singh

Microbiology Research Journal International, Page 111-116
DOI: 10.9734/mrji/2020/v30i930270

Aims: An amylase, the enzyme that catalyses the hydrolysis of starch into sugars are produced in both animals and plants and as well as in some bacteria also. Nowadays, the use of amylase enzyme in different industrial sectors and particularly in controlling the industrial water pollution has been increased as this enzyme is effective against different types of industrial effluents such as wastes from dairies, confectionaries, municipal wastes, bakery and so on which are expelled out in water without giving any proper treatment. As, the production of synthetic amylase enzyme for this purpose is quite costly, in this study, the waste water effluent was used to isolate the amylase producing bacteria, hence, decreasing the cost.

Methodology: The samples were taken from drains coming from bakery, municipal waste, etc, and five bacterias were isolated at dilution 10-6  which were named accordingly as Tan1, Tan2, Tan3, Tan4 and Tan5 further followed by Gram staining and biochemical characterization tests for further confirmation of amylase producing bacteria followed by the immobilisation of the β-amylase enzyme produced by the bacteria.

Results and Conclusion: Two bacterias were identified as amylase producing, Tan1 and Tan2, in which Gram positive bacteria showed higher amylase production at 30°C(Tan1). Compared to the free β-amylase, the immobilised β-amylase enzyme showed broader pH and temperature ranges, enhanced thermal stability, better storage stability, reusability and higher accessibility of the substrate to the immobilised β-amylase. Improved activity recovery and enzymatic properties of immobilised β-amylase chitosan beads in present study holds a promising future in industrial applications.

Open Access Review Article

Epidemiological Assessment and Challenges of the COVID-19 Pandemic in Nigeria

Stephen Olaide Aremu, Emmanuel Olumuyiwa Onifade, Babatunde Fatoke, Samuel Olusegun Itodo, Oluwatosin Oladipo, Olufisayo Bademosi, Temilade Taiwo Oleni, Tosin Adebola Ode, Ebenezer Oluwasanmi Abeleje, Adebowale Olasehinde Folorunso

Microbiology Research Journal International, Page 11-29
DOI: 10.9734/mrji/2020/v30i930259

The COVID-19 which belongs to the coronaviridae family has continued to spread in a geometric progression version. The disease that originated from Wuhan, Hubei, China has spread to all the continents of the World except Antarctica continent. As of the 5th of August, 2020 there are over 18 million reported cases of COVID-19 from 214 countries and territories of the world. More than 10 million people have recovered while approximately 696,147 people have died due to COVID-19. This review provides general information on the COVID-19 and gives deep insight into the course of the disease, interventions challenges and possible solutions in Nigeria “the giant of Africa”.

Scientific databases including Science Direct, Pub Med, Elsevier, Scopus, and Nature were explored. Data has also been accessed from case reports, newspaper reports, internet data, World Health Organisation (WHO) reports, Centre of Disease Control (CDCs) and Nigerian Centre of Disease Control (NCDCs) reports. US National Library of Medicine, Clinicaltrials.gov, has been accessed to get information about ongoing clinical trials. The literature survey started in the first week of April, 2020 and was completed in the first week of August, 2020. The clinical symptoms of COVID-19 patients are generally categorized as critical, severe, moderate and mild or even asymptomatic in descending order in terms of severity. Predictions from experts in different parts of the World concerning the possible impact of the disease in Africa have been on the downside which is due to a lot of glaring factors including poor health facilities and services.