Comparative Analysis Of Antibiotic Resistance And R-Plasmids Of Staphylococcus aureus Isolates From Human And Dog Samples

SO Akano, OA Daini, M Ojo, SI Smith, KA Akinsade



Bacterial resistance to antibiotics constitutes a major cause of failure in the treatment of bacterial infections. The genetic exchange of plasmids containing antibiotic resistant determinants between bacteria is believed to play a critical role in the evolution of antibiotics resistant bacteria and this has been shown in S. aureus. This study was therefore carried out to investigate the nature of plasmids that determine antibiotic resistance in Staphylococcus aureus isolates from man and animal. Thirty multiply drug resistant S. aureus isolates from a total of 147 apparently healthy humans and dogs, as well as from clinical cases were determined by antibiotic susceptibility test using the standard disc agar diffusion method. Plasmid isolation was carried out by the alkaline lysis method of Birnboim and Dolly. Electrophoresis as well as the transformation experiment was done.
The result showed that no particular sensitivity pattern or plasmid profile  can be ascribed to either human or animal sources of isolates. Two isolates from a domestic dog and its owner (human) were observed to have identical plasmid profile and almost the same antibiogram. 23.130 kbp and 25.119 kbp plasmids that were responsible for amoxycilin resistance were transferred. In conclusion, the genetic basis of antibiotic resistance by S. aureus in our locality was found to be partly plasmid mediated. Plasmid analysis, in conjunction with the antibiogram is valuable in differentiating multiple resistant S. aureus.
Furthermore, domestic pet animals were found to be reservoirs and potential risk factors in the transfer of multiply antibiotic resistant S. aureus and their R-plasmids to antibiotic susceptible S. aureus and other bacteria.

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Lassa Fever: Another Infectious Menace

GM Adewuyi, A Fowotade, BT Adewuyi



Nigeria is presently suffering from another Lassa fever epidemic. This was confirmed in the statement of the Minister of Health of the Federation in which he said, “There has been an upsurge in the reported cases of Lassa fever since the beginning of this year, especially in the Federal Capital Territory and its environs. Within two weeks, 12 cases with five deaths due to the disease were recorded. 25 contacts are confirmed by laboratory
investigations to have been infected, including 4 health staff working in the National Hospital, Abuja.”1 Lassa fever is an acute viral haemorrhagic fever first described in 1969 in the town of Lassa in Borno state, Nigeria.2 It is
endemic in West African countries, and causes 300,000 cases annually with 5000 deaths.3 Lassa fever epidemics occur in Nigeria, Liberia, Sierra Leone, Guinea and the Central African Republic.4 Lassa virus, the agent of the disease is a member of the Arenaviridae family. The virus is pleomorphic with single-stranded and bisegmented RNA genome.3 Its primary host is Natal Multimammate Mouse (Mastomys natalensis). Transmission to man occurs via exposure to the rat excrement through respiratory or gastrointestinal tracts5, exposure of broken skin or mucus membrane to infected material, direct contact, sexually and transplacentally. The prevalence of antibodies to the virus is 8-22%9 in Sierra Leone, 4-55% in Guinea,12 and 21% in Nigeria.13 The
disease is mild or asymptomatic in 80% of infected people, but 20% have a severe multisystemic disease. Clinical features are difficult to differentiate from that of other viral haemorrhagic fevers  and common febrile illness such as Malaria, Typhoid fever and so on. Definitive diagnosis is by viral isolation, Antigen and Antibody detection and Reverse
Transcriptase PCR. Treatment is with Ribavirin, an antiviral agent. No vaccine is currently available. Prevention is by keeping rats away from homes.

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Prevalence Of Bacterial Vaginosis In Women With Vaginal Symptoms In South Province, Rwanda

CM Muvunyi, TC Hernandez



This is a prospective study of 297 Consecutive High Vaginal Swab (HVS) specimen from patients with vaginal symptoms at the laboratory of Butare University Teaching Hospital, South Province, Rwanda. The aim of the
study was to evaluate the prevalence of bacterial vaginosis and the role of some micro-organisms and laboratory indices associated with it. The age range was 16-57 years with a mean of 30.8 years. The overall prevalence of bacterial vaginosis was 17.8% and the highest percentage of 52.8 % (28) found in the age group of 21-30 years compared with the lowest percentage of 1.9% (1) in the age group less than 20 years. Almost half of patients with trichomoniasis were found to have bacterial vaginosis (P<0.05). The demonstration Clue cells in wet mount was found in significantly higher numbers (90.5%) in women with bacterial vaginosis (P<0.001, positive predictive value 90.4%) while low sensitivity and positive predictive value were seen for vaginal discharge for detecting
infection with bacterial vaginosis ( p> 0.05, positive predictive value 26.0%). Bacterial vaginosis is common among women with vaginal symptoms in Rwanda as showed by gram stain examination. Further research into this pathology in other Rwandan women populations is needed.

Profile Of Institutional Infrastructure For Implementing Universal Precautions In Primary Health Care Facilities In Sokoto State, Nigeria: Implication For Occupational Safety

HO Isah, K Sabitu, MTO Ibrahim



Background: The adoption of primary health care in Nigeria has led to the expansion of health care delivery frontiers especially at the rural level. At this level is the most critical health services delivery point, with an attendant increase in contact between primary health care providers and patients. There is however also a simultaneous increased exposure to occupational and related health risks and hazards.Methods: The objectives of this study were to assess the universal precaution profile of primary health care facilities and determine those factors that inform their prevailing safety status. Using a structured checklist, 23 representative primary health care facilities from the 23 local government areas in Sokoto State were randomly selected for the study, one from each of the local government areas.
Results: The facilities were found to have poor universal precaution profile that could guarantee effective control of infection transmission and safety of their personnel. The facilities’ mean score on measures and frameworks for ensuring the implementation of Universal Precautions was 53.12% ± 21.68% with only 56.52% scoring above 50%.
Conclusion: Safety protocol and facilities for ensuring safe environment were inadequate and poorly developed. None of the facilities had full complement of facilities or resources for ensuring safety of working environment and for personnel’s implementation of Universal Precautions. Policy for safety practice was poor, and post exposure intervention programmes for staff in event of accidental exposure grossly underdeveloped. Interventions to improve safety environment and creation of safe climate are essential to protect primary health care workers against occupational hazards.

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Profile Of Institutional Infrastructure For Implementing Universal Precautions In Primary Health Care Facilities In Sokoto State, Nigeria Implication For Occupational Safety

Bronchopulmonary Tuberculosis- Laboratory Diagnosis And Dots Strategy Outcome In A Rural Community: A Retrospective Study

CP Enwuru, MS Emeh, IS Izuehie, SI Umeh, CA Enwuru, UM Agbasi



Pulmonary tuberculosis is still a global public health threat. Despite all efforts at its containment, the scourge is still menacing especially in the rural communities and among HIV infected patients. This retrospective study was carried out to determine the case detection rate of pulmonary tuberculosis in a rural community hospital in Nigeria from 2001-2006. A total of 1219 suspected patients were tested for pulmonary tuberculosis by sputum smear stained by Ziehl-Neelsen technique. Out of this number, 350 (28.7%) were positive for Acid-Fast Bacilli including 198 males and 152 females. Also 235 of the sputum-smear positive patients were tested for the human immunodeficiency virus (HIV) antibodies by Immunocomb 11 HIV 1 & 2 Bispot and confirmed by Immunocomb 11 HIV 1 & 2 Combfirm and HIV-1 Western Blot kit. Sixty three (26.8%) of the sputum-smear positive patients were co-infected with HIV. Two hundred and seventy (77.1%) of the AFB positive patients were treated under the Directly Observed Therapy-Short course, 201 of them (74.4%) completed the treatment, 39 (14.4%) defaulted, 30 (11.1%) died before the completion of the treatment, 195 of the patients were declared cured and 6 were declared failed. Case detection rates could be improved upon by providing culture facilities at the DOTS centers. Also efforts should be made to ensure that all positive cases are followed to a logical conclusion and that anti-retroviral drugs are provided for patients co-infected with HIV to reduce the mortality rate of pulmonary tuberculosis

The Use Of Rap-PCR In Studying Mycobacterium tuberculosis Intracellular Gene During Macrophage Infection

OAT Alli, PDF Butcher, JA Mangan, O Akinloye, CL Spreadbury



Mycobacterium tuberculosis is the second leading cause of death from infectious agent. This study sought to detect M. tuberculosis genes, which were specifically expressed, or upregulated during intracellular infection of
J774 murine macrophages; as such genes may be potential targets for novel drug action. J774 murine macrophage cell line was infected with M. tuberculosis (H37Rv strain) at 10:1 multiplicity of infection (MOI). RNA was
differentially extracted from M. tuberculosis infecting J774 macrophage cell line. The control in this case was RNA from extracellular broth grown bacteria. Approximately 50 ng of RNA from intracellular derived bacteria and extracellular derived bacteria (control) were subjected to random arbitrarily primed PCR (RAP-PCR) using 50 primer combinations. Eleven differential RAP-PCR products were observed. All RAP-PCR products were cloned into pCR®2.1 and sequenced in order to determine the identity of the products. Four of the eleven products were derived from macrophage genes and another 4 products were derived from the M. tuberculosis rRNA genes (three 23S and one 16S rRNA). The 3 remaining RAP-PCR products were found to be mycobacterial genes other than ribosomal genes. The three products were genes encoding enzyme involving in a shikimate pathway, a putative carboxyphosphonoenolpyruvate phosphonomutase and a serine protease with homology to HtrA. Of the 3 mycobacterial genes other than ribosomal genes detected, none were specifically expressed during intracellular infection but competitive RT-PCR showed that aroF gene was upregulated two-fold in intracellular derived bacilli.

Optimization Of Rna Extraction In Mycobacterium Tuberculosis For Studying Intracellular Gene Expression

AOT Alli, JA Mangan, PD Butcher, CL Spreadbury



Mycobacterium tuberculosis is the leading cause of death due to infectious disease after Human immunodeficiency virus. There has been an upsurge in the incidence of tuberculosis since 1980s. In order to reverse this trend, there is need to understand the biology of the organism. This can be brought about by studying gene expression at transcriptional level. The success of this hinges on RNA of good quality. In this paper, five methods (hot phenol, sonication with guanidinium thiocyanate (GTC) solution, beadbeating method with Trizol, FastPrep machine with Divolab as detergent and GTC solution, and FastPrep machine with Trizol) of extracting RNA from bacteria were compared to find
which of the method would be suitable for mycobacteria. The study found that physical method of lysing bacteria was necessary for extraction of RNA from mycobacteria. FastPrep machine gave the highest yield and also provided the speed necessary for optimum RNA extraction. FastPrep and Trizol as reagent for extraction of RNA was applied to macrophage infected with M. tuberculosis (H37Rv) after removing the macrophage RNA. We were able to demonstrate the expression of dnaK gene in both intracellular and broth grown bacilli. The expression of dnaK gene was found to be downregulated in macrophage compared to broth.

African Journal of Clinical and Experimental Microbiology Vol. 10 (2) 2009: pp. 64-79

Studies On The Distribution Of Clinically Diagnosed Pulmonary Tuberculosis In Ebonyi State, Nigeria

RK Obi, AN Amadi, IM Idika, FC Nwanebu



A study was conducted between January, 2005 to December, 2006 to ascertain the prevalence of pulmonary tuberculosis among patients who attended chest clinics in some randomly selected hospitals, Clinics and Health Centers in Ebonyi State, Nigeria. Investigations were carried out using the two popular diagnostic criteria for pulmonary tuberculosis namely the specific Ziehl Neelsen (ZN) methods and chest X-ray. A total of 962 patients with clinical signs and symptoms of tuberculosis were studied. Out of this figure, 559 (58.1%) had pulmonary tuberculosis with 1:1 male/female ratio. There was no significant difference between ZN sputum smear positive and chest x-ray in the diagnosis of pulmonary tuberculosis in the studied population at 95% confidence level. The possible reasons for the high prevalence of pulmonary tuberculosis may be attributed to increase in the incidence of HIV/AIDS, high rate of poverty, emergence of drug-resistant strains of Mycobacteria and to a lesser extent, smoking and diabetes

Keywords: Pulmonary tuberculosis, sputum, chest X-ray, Ziehl Neelsen

African Journal of Clinical and Experimental Microbiology Vol. 10 (2) 2009: pp. 80-87

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Sensitivities Of Citrobacter, Proteus And Providencia Isolates To Sulbactam-ampicillin, Trimethoprim-S Ulfamethoxazol And Ticarcillin-Clavulanic Acid Antibiotics

G Uraz, KOK Turhan, HD Binnet



Gram negative bacterias which belong to Enterobacteriaceae family which is critically important as a matter of human health, are comperatively prevalent in nature and foods. Infections formed by bacterias resistant to antibiotics significantly cause mortality and economical losses. Sensitivities of gram-negative bacterias isolated from miscellaneous samples to sulbactamampicillin, trimethoprim-sulfamethoxazol and ticarcillin-clavulanic acid were analysed. In this study, Proteus is primary isolated microorganism with % 52,08. It was determined that Proteus types are proportionally 86,66% sensitive to Trimethoprim-Sulfamethoxazol and 76% resistant to Sulbactam-Ampicillin . Citrobacter is the secondary isolated microorganism (31,25%). It was determined that Citrobacter types were 86,66% sensitive to Trimethoprim-Sulfamethoxazol and % 73,3 resistant to Sulbactam-Ampicillin. Providencia types which are thirdly isolated microorganism (16%), For Providencia types, Trimethoprim-Sulfamethoxazol were determined as the most effective antibiotic again with 86,66% sensitivity. Resistance to Sulbactam- Ampicillin was however, found to be 76%. In conclusion, it is very important to conduct sensitivity tests in choosing antibiotics for chemotherapy of infections. Trimethoprim-Sulfamethoxazol is recommended in the empiric treatment of urinary tract infections in our environment

Keywords: Enterobacter, sensitive, SXT, TĐM, SAM

African Journal of Clinical and Experimental Microbiology Vol. 10 (2) 2009: pp. 88-91

Candida Species Amongst Pregnant Women In Benin City, Nigeria: Effect Of Predisposing Factors

FE Oviasogie, FI Okungbowa



Genital samples from pregnant women were examined to determine the Candida species present and how some predisposing factors would affect the frequency of isolation of species. A total of 147 women (87 volunteer asymptomatic pregnant women and 60 asymptomatic nonpregnant women) were examined. High Vaginal Swabs (HVS) were collected in pairs while demographic data (age of individual, age of pregnancy, occupation and use of contraceptive pill) on each subject were obtained alongside. Samples were cultured on Sabouraud Dextrose Agar plates and incubated for 72 hours at 37°C. Results revealed five Candida species (C.albicans, C.glabrata, C.tropicalis, C.stellatoidea, C.parapsilosis) with 61.5, 17.9, 7.7, 5.1, 2.6 % frequency of isolation respectively, as against 73.7, 7.7, 1.4, 11.3, and 0.0% for control. Women in the second trimester of pregnancy had the highest occurrence (68.8%) while the age groups 24- 30 and 31-37 years had the highest occurrence in the control (46.2%) and in the pregnant ones (51.7%) respectively. Traders had 68.8%, and full-time housewives, 26.7% occurrence. More women who used contraceptives had Candida (58.3%) as against non contraceptive users (35.3%). The results support literature reports that there are high rates of colonization of the vagina of pregnant women by Candida. They also give credence to earlier reports that the factors determined here affect the frequency of isolation of Candida species from the vagina.

KeywordsCandida, pregnancy, women, genital specimens, Benin City

African Journal of Clinical and Experimental Microbiology Vol. 10 (2) 2009: pp. 92-98