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

CA Enwuru, A Ogunledun, N Idaka, NV Enwuru

 

Abstract

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 < 12mm) using agar diffusion method, matched with 58(78.4%) sensitive MIC < 8μg/ml, 9 (12.2%) S-DD MIC 16-32μg/ml and 7(9.5%) resistant MIC >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. 88-96

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Opportunistic infections and clinico-epidemiological factors in HIV/AIDS casesseen in a tertiary care hospital in Nepal

RN Das, HS Joshi, R Biswas, HS Joshi

 

Abstract

Opportunistic infections are the leading cause of morbidity and mortality among HIV/AIDS patients. The spectrum of opportunistic pathogens involved in such infections in Nepal is not well documented. A cross sectional (hospital-based) study was carried out at the AIDS clinic of Manipal Teaching Hospital, Pokhara, Nepal. A total of 404 clinically suspected cases of HIV/AIDS seen at the clinic between July 2001 and December 2002, were screened for HIV. Seventy four (18.3%) were sero-positive for HIV. Fever was the commonest presenting symptoms 48.6% followed by cough and dyspnoea 36.5%, weight loss 36.5% and pulmonary tuberculosis 21.6%. Fifty five of the 74 (74.3%) HIV positive cases were in the age group 20-39 years. Heterosexual mode of acquisition/transmission was seen in 60.8% and 21.6% were intravenous drug abusers (IVDA). A total of 45 opportunistic pathogenic isolates were recovered from the 74 patients. Mycobacterium tuberculosis was the commonest pathogen 60%, followed by Cryptosporidium spp 13.3% and Candida spp 11.1%. Four patients died during the period of study giving a mortality rate of 5.4%. This study shows that HIV/AIDS is rapidly becoming a grave concern in the Pokhara valley of Nepal. Intensive and effective health education programmes among the target population may be a cost effective method to curb the rising prevalence of HIV/AIDS in a developing country like Nepal. Also, further regional studies are required to establish more detailed epidemiological database of opportunistic infections in HIV/AIDS patients in Nepal.

Keywords: HIV/AIDS, opportunistic infections, Nepal

African Journal of Clinical and Experimental Microbiology Vol. 6 (3) 2005: 239-245

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OSMOTIC FRAGILITY AND Na+ -K++ ATPase ACTIVITY OF ERYTHROCYTES OF HIV/AIDS PATIENTS

OAT Ebuehi, M Balogun, RA Audu, OE Idigbe

 

Abstract

A cross sectional study was carried out to investigate the osmotic fragility and Na+ -K+ ATPase activity of the erythrocytes of HIV/AIDS patients. Whole blood was taken from subjects at the Human Virology Laboratory of the Nigerian Institute of Medical Research. Subjects were judged suitable for the various investigations by means of a questionnaire. The Genie II HIV diagnostic kit was used to confirm HIV positive status. HIV positive subjects were grouped into two: those receiving anti-retroviral therapy were referred to as the ARV group and those not receiving antiretroviral therapy were designated as non-ARV group. Each group was further sub-divided according to the Centers for Disease Control 1993 classification of HIV disease. HIV negative subjects must have tested no later than two months to the sample collection date and must not lead a high-risk lifestyle. Twenty microliters of whole blood were used for the erythrocytes osmotic fragility assay. One milliliter of whole blood was used to prepare the erythrocyte ghost membrane for the Na+-K+ ATPase activity assay. The two HIV positive groups showed significant increase in percentage haemolysis under osmotic stress at 0.65% saline. The ARV group had an average percentage haemolysis of 2.56 ± 0.81% while the non-ARV group had an average of 3.19 ± 1.11% compared to an average of 0.83 ± 0.36% for the control group (p < 0.05). A pattern observed in the result was an increase in activity with increasing severity of the HIV/AIDS disease. Data from the present study indicate that the osmotic fragility of erythrocytes was significantly potentiated, while Na+-K+ ATPase activity was not significantly altered (p < 0.05) in HIV/AIDS disease.

Key Words: Osmotic fragility, Na+ -K+ ATPase activity, erythrocytes, HIV/AIDS

Afr. J. Clin. Exper. Microbiol. 2004; 5(2): 148 – 154.

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BLOOD CHEMISTRY AND PLATELET SEROTONIN UPTAKE AS ALTERNATIVE METHOD OF TRACKING HIV/AIDS

OAT Ebuehi, M Balogun, RA Audu, OE Idigbe

 

Abstract

A cross sectional study was conducted to investigate the blood chemistry and platelet serotonin uptake as alternative method of determining HIV disease stage in HIV/AIDS patients. Whole blood was taken from subjects at the Human Virology of the Nigerian Institute of Medical Research. Subjects were judged suitable for the various investigations by means of a questionnaire. The Genie II HIV diagnostic kit was used to confirm HIV positive status. HIV positive subjects were grouped in to two: those receiving antiretroviral therapy were referred to as the ARV group and those not on antiretroviral therapy were designated as non-ARV group. Each group was further sub-divided according to the Centers for Disease Control 1993 classification of HIV disease. HIV negative subjects must have been tested no later than two months to the sample collection date and must not lead a high-risk lifestyle. Serum was used to assay for blood chemistry activities with Randox analytical reagents. Blood platelets were prepared from one milliliter of whole blood and platelet serotonin uptake rates were determined. The serum glutamic-oxaloacetic transaminase (SGOT) of non-ARV subjects was the only blood chemistry parameter that showed any significant variation from normal (p<0.05). The mean activity of this enzyme was 28.4 ± 5.29 U/L compared to a normal value of 12 U/L. A disease stage-related variation was observed. Platelet serotonin uptake rates of the two HIV positive groups showed no significant difference with the HIV negative control. The data obtained showed that serum glutamic-oxaloacetic transaminase activity is significantly increased in HIV/AIDS patients in a manner that is disease stage related. However, serum glutamic-pyruvic transaminase, bilirubin, triglycerides, amylase, serum creatinine, and alkaline phosphatase showed no significant variation from normal values. Platelet serotonin uptake of HIV subjects was not significantly different from the control.

Key Words: Blood chemistry, platelet serotonin uptake, HIV/AIDS

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BLOOD CHEMISTRY AND PLATELET SEROTONIN UPTAKE AS ALTERNATIVE METHOD OF TRACKING HIVAIDS

Afr. J. Clin. Exper. Microbiol. 2004; 5(2): 155 – 159