Susceptibility profile of yeast-like organisms isolated from HIV/AIDS patients; using NCCLs macrodiltion method compared with agar diffusion technique.

Yeast like opportunistic fungal infection has been reported globally amongst HIV/AIDS patients, particularly as the etiologic agent of oral thrush. Fluconazole antibiotic has been most popularly employed in treating cases of oral thrush in HIV/AIDS patients. Recent reports have recorded antifungal drug resistance amongst immunocompromised subjects. This constitutes a big problem in the management of opportunistic candidiasis. The NCCLS micro/macrodilusion sensitivity testing procedure is expensive, cumbersome and requires a level of sophistication. This study was designed to compare NCCLS M-27-A macrodilution method (expensive) with agar diffusion technique (cheap and simple), to provide a reliable rapid alternative to the new pressing need for antifungal routine sensitivity testing. Sputum specimens from 213(108 females and 105 males) HIV positive patients were plated onto SDA. The isolates were identified by morphotyping, microscopy and speciated using germ tube test, and battery of biochemical sugar fermentation; and assimilation tests. Fluconazole agar diffusion susceptibility testing was carried out on each isolate, compared with the NCCLS macrodilution sensitivity assay standard. Of the 74 isolates tested for fluconazole sensitivity, 59(79.7%) were sensitive (zone diameter > 19mm, mean diameter 28mm), 6(8.1%) were Sensitive Dose Dependent (S-DD) (zone diameter 13-18mm, mean diameter 16mm), while 9(12.2%) were resistant (zone diameter 64μg/ml profile, using the NCCLS macrodilution assay. The differences between the test method (Agar diffusion) and the control standard method (NCCLS-M 27-A broth Macrodilution MICS) were not statistically significant using t-test (two tail) (t = 4.302656, P=1.0). Among the C. albicans isolates, 26(86.7%) were sensitive to fluconazole. The rank of susceptibility was C. albicans > C. tropicalis > C. krusei. It is concluded that broncho-oro-pharyngeal Candida and other yeast-like species existed in about one third of the HIV and AIDS patients studied; in which C. albicans was the most prevalent, while about ten percent of all the Candida isolates were resistant to fluconazole. The reliability of germ tube production as a confirmatory test for Candida albicans in HIV infection was as high as 96.7% and is therefore, recommended for continued use. Agar diffusion compared favourably with the NCCLS macrodilution technique, hence it is recommended for routine antifungal sensitivity test on all isolates of yeast-like cells from HIV and AIDS subjects. Keywords: HIV/AIDS, oral thrush, yeast-like cells, fluconazole resistance, NCCLS vs agar diffusion technique.African Journal of Clinical and Experimental Microbiology Vol. 9 (2) 2008 pp. 88-9

A study of microbial analysis of fresh fruits and vegetables, in Sagamu markets South-West, Nigeria

Raw and minimally processed fruits and vegetables are essential parts of diets of people around the world. However, these food items have consistently served as vehicles for human diseases worldwide. This study was aimed at determining the microbiological quality of fresh fruits and vegetables sold at Sagamu markets, Ogun state, Nigeria. Twenty different samples of fruits and vegetables were randomly purchased from ten various vendors in Shagamu market, Ogun State, Nigeria. Aerobic plate counts, total coliform counts, yeast, mold counts and antimicrobial susceptibility study using were determined using standard microbiological methods. In Fruits, the isolation rate of Bacillus specie was predominant (44.1%), followed by Klebsiella species (44.0%) while in vegetables, the prevalent rates of 38.3% and 14.4% were respectively recorded by Staphylococcus aureus and Pseudomonas species. Malassezia species and Aspergillus species were the only fungal isolates obtained from the fruits and vegetables. Aspergillus species were predominant in vegetables, with a frequency of 13.3% while both Malassezia and Aspergillus species were observed almost at the same frequently of 24.1% and 24.4% in fruits respectively. This study showed that fruits and vegetable were contaminated with potential food borne pathogens with varying degree of antibiotic resistance thus suggesting that they could act as a vehicle for the dissemination of antibiotic resistant organisms as well as serve as a reservoir of food borne pathogens of public health significance.Keywords: Micro flora, fresh fruits and vegetables, antibiotics resistanc

Growth and survival of Shigella flexneri in commonly consumed foods in Nigeria

The incidence of shigellosis is on the increase in Nigeria and foods have been strongly implicated as vehicles of transmission of infection. In this study the survival and growth of Shigella flexneri were investigated in foods that are commonly consumed in the country which include cooked rice, beef, cooked beans, fish, cooked maize and banana (Musa sapientum). Survival at 5°C and growth at 25°C and 37°C were quantitated by viable cell counts on MacConkey agar. The organism survived and showed considerable growth in all the foods examined. Cell growth increased from 105 to 1010 within 6 and 24h following inoculation at 25°C and 37°C. Keywords: foods, survival, Shigella flexneri, shigellosis, Nigeria Nigerian Food Journal Vol. 23 2005: 205-20

Quinolones resistance and R-plasmids of some gram negative enteric bacilli

Out of the two hundred and sixty bacteria isolates from clinical specimens obtained from different body sites at the University College Hospital Ibadan, 166 belonged to the family of Enterobacteriaceae and Pseudomonaceae. The isolated gram-negative enteric bacilli consist of Escherichia coli (22), Klebsiella species (65), Proteus species (20), Salmonella typhi (2), Pseudomonas aeruginosa (39) and Pseudomonas species (18). Among the antimicrobial agents tested, high resistance was found with ofloxacin 44.0%, followed by pefloxacin 30.1% and ciprofloxacin 21.7%. Ciprofloxacin has the lowest MIC of 2 – 32 μg/ml while ofloxacin has the highest 64 μg/ml. Of the 166 strains, 44 were resistant to most of the antimicrobial agents tested. All the strains that were resistant to any antimicrobial agents were also resistant to ofloxacin. A total of 27 plasmids ranging in molecular sizes from 6.6.kb to 17.4kb were extracted from the resistant strains and grouped into 5 plasmid profiles. Transformation experiment revealed that 59.2% of the resistant strains carried a common R-plasmid of size 10.7kb. Plasmid-mediated resistance to ciprofloxacin and pefloxacin was found. Klebsiella species harboured the highest number of R-plasmids with 8, followed by Pseudomonas aeruginosa with 4. Afr. J. Clin. Exper. Microbiol. Vol.6(1) 2005: 14-2

Comparative kinetics of digoxin sensitive (NAK) ATPase and Ca 2+ ATPase in symptomatic and asymptomatic patients with faciparum malaria

No Abstract. Journal of Malaria in Africa and the Tropics Vol. 1 (2) 2002: pp. 21-2

High levels of multidrug resistance in clinical isolates of Gram-negative pathogens from Nigeria.

International audienceIn Nigeria, quinolones and β-lactam antibiotics are widely used to treat bacterial infections. This study aimed to identify the prevalence of resistance to these drugs and to determine the mechanisms of resistance to these agents. In total, 134 non-duplicate, Gram-negative enteric isolates of 13 species from different hospitals were investigated for susceptibility to a panel of antibiotics, carriage of plasmid-mediated quinolone and β-lactam resistance genes, production of extended-spectrum β-lactamases (ESBLs), and mutations within topoisomerase genes. The level of resistance to all antibiotics tested was extremely high, with minimum inhibitory concentrations for 90% of the organisms (MIC values) of ≥256μg/mL for all drugs. Of the 134 isolates, 92 had mutations within the quinolone resistance-determining region (QRDR) of or within and . In addition, the plasmid-mediated quinolone resistance genes , , and were identified. The allele, which has previously only been found in isolates from China, was identified in two isolates and one isolate. Of the 134 isolates, 23 (17.2%) carried , 11 (8.2%) carried a variant and 5 (3.7%) were positive for . Twenty-eight isolates (20.9%) produced ESBL variants, with a CTX-M variant being carried by 25 isolates (18.7%). In addition, six isolates (4.5%) carried variants [ACT-1 (1 isolate), DHA-1 (4 isolates) and CMY-2 (1 isolate)]. This study demonstrates a very high level of multidrug resistance amongst Gram-negative enteric bacilli isolated from different sites from patients in Nigerian hospitals as well as the presence of a variety of plasmid-associated resistance genes, including some identified from Africa for the first time

Fluconazole resistant opportunistic oro-pharyngeal candida and non-candida yeast-like isolates from HIV infected patients attending ARV clinics in Lagos, Nigeria.

Background: Oro-Pharyngeal Candidiasis (OPC) continues to be considered the most common opportunistic fungal disease in HIV/AIDS patients globally. Azole antifungal agent has become important in the treatment of mucosal candidiasis in HIV patients. Presently, antifungal drug resistance is fast becoming a major problem particularly with the immune depleted population. Objectives: This study was designed to investigate the: existence of OPC, species distribution fluconazole susceptibility profile of yeast cells isolated from oral specimens of HIV/AIDS patients from Lagos Nigeria, between Oct. 2004 and June, 2005. Methodology: The venous blood samples were screened for HIV antibodies using the Cappillus HIV I and II test kit (Trinity Biotech Plc UK), and Genie II HIV I and II EIA kit (Bio-Rad France). The positive results were subsequently confirmed at the laboratory attached to each of the clinics, using the Nigerian Federal Ministry of Health approved algorithm. The samples from 213 (108 females and 105 males) HIV positive patients were plated onto SD agar. The isolates were identified by morphotyping, microscopy and speciated using germ tube test and battery of biochemical sugar fermentation and assimilation tests. Fluconazole agar diffusion susceptibility testing was carried out on each isolates. Results: Seventy-four (34.7%) isolates were recovered including one person with double isolates. Only 70(94.6%) of the isolates could be adequately speciated. Candida albicans 30 (40.5%) was the most frequently isolated species, the rest were non-albicans species, with the frequency of C. tropicalis › C. Krusei › C. glabrata and C. neoformans for species for species having up to 4 isolates. Four (30.8%) out of 13 isolates of C. tropicalis showed germ tube formation. While one C. albicans was germ-tube negative. Out of the 74 isolates tested for fluconazole sensitivity, 58(78.4%) were sensitive, MIC d” 8μg/ml, 9(12.1%) were susceptible Dose Dependant (SDD), MIC 16-32 μg/ml and 7(9.5%) were resistant, MICs e” 64μg/ml. Among the C. albicans isolates, 26(86.7%) were sensitive to fluconazole. The rank of susceptibility was C. albicans > C. tropicalis > C. Krusei for the most prevalent species. Conclusion: We conclude that fluconazole resistant strains of oro-pharyngeal yeast-like cells exist in about 9.5% of HIV/AIDS patients with the above stated species distribution. We therefore, highlight the need for routine antifungal susceptibility testing on HIV patients with cases of initial or repeat episodes of OPC. Keywords: Oropharyngeal Candida (yeast-like cells), HIV/AIDS and Fluconazole Resistance. African Health Sciences Vol. 8 (3) 2008: pp. 142-14

Fluconazole resistant opportunistic oro-pharyngeal candida and non-candida yeast-like isolates from HIV infected patients attending ARV clinics in Lagos, Nigeria

Background: Oro-Pharyngeal Candidiasis (OPC) continues to be considered the most common opportunistic fungal disease in HIV/AIDS patients globally. Azole antifungal agent has become important in the treatment of mucosal candidiasis in HIV patients. Presently, antifungal drug resistance is fast becoming a major problem particularly with the immune depleted population. Objectives: This study was designed to investigate the: existence of OPC, species distribution fluconazole susceptibility profile of yeast cells isolated from oral specimens of HIV/AIDS patients from Lagos Nigeria, between Oct. 2004 and June, 2005. Methodology: The venous blood samples were screened for HIV antibodies using the Cappillus HIV I and II test kit (Trinity Biotech Plc UK), and Genie II HIV I and II EIA kit (Bio-Rad France). The positive results were subsequently confirmed at the laboratory attached to each of the clinics, using the Nigerian Federal Ministry of Health approved algorithm. The samples from 213 (108 females and 105 males) HIV positive patients were plated onto SD agar. The isolates were identified by morphotyping, microscopy and speciated using germ tube test and battery of biochemical sugar fermentation and assimilation tests. Fluconazole agar diffusion susceptibility testing was carried out on each isolates. Results: Seventy-four (34.7%) isolates were recovered including one person with double isolates. Only 70(94.6%) of the isolates could be adequately speciated. Candida albicans 30 (40.5%) was the most frequently isolated species, the rest were non-albicans species, with the frequency of C. tropicalis > C. Krusei > C. glabrata and C. neoformans for species for species having up to 4 isolates. Four (30.8%) out of 13 isolates of C. tropicalis showed germ tube formation. While one C. albicans was germ-tube negative. Out of the 74 isolates tested for fluconazole sensitivity, 58(78.4%) were sensitive, MIC d" 8µg/ml, 9(12.1%) were susceptible Dose Dependant (S-DD), MIC 16-32 µg/ml and 7(9.5%) were resistant, MICs e" 64µg/ml. Among the C. albicans isolates, 26(86.7%) were sensitive to fluconazole. The rank of susceptibility was C. albicans > C. tropicalis > C. Krusei for the most prevalent species. Conclusion: We conclude that fluconazole resistant strains of oro-pharyngeal yeast-like cells exist in about 9.5% of HIV/AIDS patients with the above stated species distribution. We therefore, highlight the need for routine antifungal susceptibility testing on HIV patients with cases of initial or repeat episodes of OPC

The patterns of Osmotic Fragility and Thrombocytopenia in Nigerian Children with Acute Plasmidium Falciparum Malaria before and after Chemotherapy

The clinical and case management application of erythrocyte osmotic fragility and peripheral blood platelets were investigated in 40 healthy children (N=40; M/F = 20/ 20 mean age 6.2 + 3.7 years) and103 falciparum malaria children with severe anaemia (N = 41; M/F=22/19; mean age = 5.3 + 1.5) cerebal malaria (N = 16; M/F = 10/6; mean age = 3.8 + 0.7 ) and uncomplicated malaria (N =46; M/F = 21/25; mean age = 7.4 + 1.3). At presentation, thrombocytopenia (platelet count 0.05) from that of healthy children. Thrombocytopenia also reduced substantially from 100% to 14.30% in cerebral malaria, 34.1% to 7.30% in severe anaemia and 17.4% to 4.30% in uncomplicated malaria. It was concluded that effective management of acute P. Falciparum malaria in Nigerian children is a combination of good therapeutic response and substantial recovery from acute illness, which can be adequately assessed by monitoring parasite clearance; osmotic fragility and platelet count patterns