Open Access Original Research Article

Detection of Extended-spectrum β-lactamases’ (ESBLs) Resistance among Urinary Tract Pathogens in Khartoum State

Eman Fadelalla Fadel Elmula, Wafa Ibrahim Elhag

Microbiology Research Journal International, Page 618-623
DOI: 10.9734/BMRJ/2015/17167

Aims: The aim of this study is to isolate and identify the extended spectrum beta lactamase (ESBLs), the causative agents of urinary tract infection and detection of their resistance against β lactam drugs.
Study Design: Descriptive cross sectional studies in which 100 patients with UTI.
Place and Duration of Study: This study was conducted at the Department of Microbiology, Faculty of Medical Laboratories Science, Al-Neelain University, Khartoum – Sudan from “1st September to 31thDecember 2014.
Methodology: One hundred urine samples were collected from Khartoum state Hospitals and identified on the basis of their culture characteristics and morphological appearance using Gram stain technique and biochemical tests. The isolates were subjected to antimicrobial susceptibility against the third generation cephalosporins (Cefotaxime, Ceftazidime and Ceftriaxone) using Disk-Diffusion method. The bacterial isolates were inoculated to show their ability to produce ESBL using Combination Disk Method (Calvulanic acid + Third generation cephalosporins). The ESBL producers were evaluated among non-ESBL producers.
Results: The isolates were identified and enumeratetd as E. coli (49), Proteus species (15), Klebsiella pneumoniae (18), Pseudomonas aeruginosa (9), Enterococcus faecalis (7) and 2 Gram positive bacteria (Staphylococcus species). The isolates subjected were found to show their antibacterial susceptibility against third generation cephalosporins (Cefotaxime, Ceftazidime, and Ceftriaxone) and the results observed were that sixty out of one hundred isolates depicted resistance to third generation cephalosporins and it included E. coli (29), Klebsiella pneumoniae (18), Proteus species (7), Pseudomonas aeruginosa (4) and Enterococcus faecalis (2). Among sixty bacterial species that showed resistance to the third generation cephalosporins, E. coli 20(68%), Klebsiella pneumoniae 7(38%), Proteus species 3(42%) and Enterococcus faecalis 1(50%) respectively were found to be ESBL producer. Pseudomonas aeruginosashowed non-ESBL producers.
Conclusion: We conclude that the ESBL producers were found in large proportion which may be due to the misuse of antibiotics or inadequate treatment.

Open Access Original Research Article

Anti-Shiga Toxin Producing Escherichia coli O157:H7 Effect of Ocimum basilicum L. Essential Oil Analyzed Using Time Kill Assay in a Batch Culture

Haidar Kadum Yakob

Microbiology Research Journal International, Page 624-634
DOI: 10.9734/BMRJ/2015/17717

Essential Oil extract from Ocimum basilicum (Labiatae; basil) was investigated for its in vitro antibacterial properties against an emerging human food-borne pathogen, Shiga Toxin Producing E. coli O 157:H7. Agar disc diffusion method and time-kill assay in a batch culture were employed. The results showed that essential oil of basil was effective to inhibit the growth of pathogenic bacteria tested. The diameter of inhibition zone was 15.25±0.58 mm. In a batch culture, four oil concentrations (4, 2, 1, 0.5, 0.25 and 0.125 mg/ml) were initially tested to determine the concentration of basil oil that could inhibit growth of E. coli O157:H7. It was found that the oil at 4, 2, 1 and 0.5 mg/ml killed the bacteria after 1 h, while at 0.25 mg/ml the number of bacteria was only reduced. Two oil concentrations, 0.25 mg/ml and 0.5 mg/ml, were chosen to study its effect on E. coli O157:H7 at different growth phases, i.e. exponential, late exponential and stationary phase in a batch culture. Oil at both concentrations (0.25 mg/ml and 0.5 mg/ml) could not completely kill the cells at the exponential phase, and upon prolonged incubation, resistant cells emerged. But basil oil at 0.5 mg/ml could completely killed cells at the late exponential and stationary phases suggesting that the oil at 0.25 mg/ml was bacteriostatic, while at 0.5 mg/ml was bactericidal against E. coliO157:H7.

Open Access Original Research Article

Virulence of Fusarium oxysporum on Kidney Organ Using In vivo Mice Model

I. P. Udoh, C. I. Eleazar

Microbiology Research Journal International, Page 635-643
DOI: 10.9734/BMRJ/2015/18012

Aim: To determine the virulence of Fusarium oxysporum on mice kidney organ.
Study Design: Study was carried out using experimental design.
Place and Duration: The study was carried out at University of Nigeria, Enugu Campus. From May 2012–August 2014.
Methodology: Isolates from human infections were obtained from 2000 patients with apparent signs of fungal infections (mycotic keratitis). Plant isolates were obtained from symptomatic plants (decaying or rotting plant substrate) from five different markets and farm lands in Enugu state, Nigeria. A total of 3000 plants were used. Cultures were carried out on Sabouraud Dextrose Agar slants according to modified methods and identifications made. In vivo virulence studies in mice were carried out with each individual strain. Histopathology of the kidney organ was performed. Mean survival time (MST) was estimated by the Kaplan-Meier method.
Results: In vivo virulence studies of Fusarium oxysporum on mice kidney organ resulted in disseminated infection and death of the animals. Histopathological analysis revealed infected kidney with evidence of intense inflammation involving the glomeruli and severe, extensive as well as severe diffuse inflammatory cell infiltration in the interstitium of the cortex. Kidney of control mice showed no pathological features.
Conclusion: The various pathological manifestations and physical signs of infection in the injected mice were indications that Fusarium oxysporum isolated were virulent proving the hypothesis that a single strain of Fusarium oxysporum can produce disease both in plant and human hosts.

Open Access Original Research Article

Ebselen a Seleno-organic Molecule Inhibits Alteration in the Biological Responses in Hypoxic Human Alveolar Lung Epithelial Cells

Neetu Kushwah, Amit Prabhakar, Dipti Prasad, K. Ravi, Shashi Bala Singh, Nilofar Khan

Microbiology Research Journal International, Page 644-654
DOI: 10.9734/BMRJ/2015/17639

Ebselen is a lipid-soluble seleno-organic compound whose benefits have been shown in a variety of diseases and in experimental studies including anti-inflammation, anti-proliferation, anti-angiogenesis, anti carcinogenesis and anti-oxidation properties. This study aimed to evaluate the effect of ebselen against hypoxia-induced biological responses in human alveolar epithelial cells (A549 cells). Hypoxia treatment increased the generation of reactive oxygen species, proinflammatory cytokines/chemokines, cell death and proliferation in A549 cells in a time-dependent manner. Notably, Ebselen treatment significantly reduced the production of reactive oxygen species and levels of biochemical markers for lipid peroxidation. Ebselen treatment increased the antioxidant enzyme superoxide dismutase activity as well. This seleno-organic compound also reduced the production of proinflammatory chemokines IL-8 and proinflammatory cytokine TNF-α. It is worth noticing that, Ebselen enhanced the viability as well as proliferation of hypoxic A549 cells. Collectively, these data demonstrate that the Ebselen attenuates the alterations in biological responses in hypoxic human alveolar epithelial cells. Our results further suggest that that ebselen could be used as a potential chemopreventive compound to restore the normal biological response in hypoxic cells and thus inhibiting the progression of the lung carcinoma.

Open Access Original Research Article

Status of Coffee Leaf Rust Resistance on Kenyan Commercial Resistant Cultivars

S. G. Ligabo, E. K. Gichuru, O. Kiplagat, B. M. Gichimu

Microbiology Research Journal International, Page 655-662
DOI: 10.9734/BMRJ/2015/16968

Background: Coffee Leaf Rust (CLR) is a fungal disease caused by Hemileia vastatrix Berk. and Br. The pathogen is constantly evolving leading to rapid break down in resistance of once resistant coffee varieties. To date, more than 49 races of the pathogen have been characterized all over the world and new races are continuously being characterized some of which are able to infect Robusta derivatives.
Aim: The objective of this study was therefore to re-examine the status of CLR resistance on Kenyan commercial resistant cultivars and investigate the pathogenic interaction between H. vastatrix isolates and their host genotypes.
Methodology: Hemileia vastatrix isolates were collected from naturally infected leaves of the host coffee genotypes and were inoculated on one Robusta coffee genotype and eight Arabica genotypes comprising of three Kenyan commercial cultivars (SL28, Ruiru 11 and Batian) and five museum genotypes (HDT, Mundo Novo, Pretoria, 110/2 and Bourbon) using leaf disks inoculation method. An infection scale of 1-6 was used to score the virulence of the pathogen isolates.
Results: There was significant variation among isolates on their virulence against the different genotypes. SL28, Pretoria and Mundo Novo were the most susceptible to most isolates while none of the isolates infected Ruiru 11, HDT and Robusta. All the isolates were able to infect Batian but none reached the stage of sporulation. Isolates from K7, SL34 and SL28 were found to be more virulent than those from Batian and Blue Mountain. Unlike the host genotype, the region from which the isolates were obtained was not found to play any role on the virulence of the isolates.
Conclusion: Although six additional races of H. vastatrix were recently detected in Kenya some of which are able to infect Robusta derivatives, the study confirmed that Kenyan genotypes in this group are still resistant against most races of H. vastatrix in Kenya. It was therefore deduced that either these new races are not yet wide spread in all coffee growing areas in Kenya or that there are other major and minor genes conditioning the coffee-rust interactions besides the SH genes.

Open Access Original Research Article

Multidrug Resistance of Salmonella Isolated from Shellfish Samples, Morocco

Rachid Boutaib, Brahim Bouchrif, Mohammed Abid, Bouchra Karraouan, Mohammed Khaddor, Amin Laglaoui

Microbiology Research Journal International, Page 663-669
DOI: 10.9734/BMRJ/2015/18501

Aims: This study aims to determine prevalence of Salmonella in shellfish and to study their resistance to antibiotics.
Samples: Three species of shellfish consisted of cockles, clams and mussels were sampled monthly in six sites during two years 2008 and 2012.
Methodology: As many as 272 samples of shellfish were examined for presence of Salmonella. Positives strains were confirmed for presence of invA gene, serotyping and tested for drug susceptibility.
Results: Up to 7.7% of samples were positive for Salmonella and a total of 90 Salmonella isolates belonging to 4 serovars (S. Kentucky, S. Glostrup, S. Newport and S. Reading) were tested for their susceptibility to a panel of 12 antimicrobial agents. Many resistant isolates were detected with 75.8% of isolates resistant to at least one antimicrobial agent. Isolates demonstrated resistant to streptomicin, chloramphenicol, nalidixic acid, ciprofloxacin, amoxicillin, respectively with (90.6%; 51.6%; 31.6%; 30.5% and 19%). The most common pattern of multiple drug resistance included resistance to chloramphenicol, ciprofloxacin, nalidixic acid.
Conclusion: It seemed that strains isolated of Salmonella were multidrug resistant and almost one third of strains were resistant to quinolone.
The results emphasize the need of a monitoring programme of bacterial pathogenes Salmonella on shellfish to protect human health.