Correction: HIV-1C proviral DNA for detection of drug resistance mutations.

[This corrects the article DOI: 10.1371/journal.pone.0205119.].

HIV-1C proviral DNA for detection of drug resistance mutations.

BACKGROUND: Using HIV proviral DNA as a template may be suitable for initial detection of transmitted drug resistance mutations (TDRMs) as it is easy to handle and less expensive compared to RNA. However, existing literatures which are mainly focused on HIV-1B subtypes DNA extracted from PBMCs revealed controversial findings ranging from the detection of significantly lower or higher mutations in proviral DNA compared to historic viral RNA. Thus, to verify whether viral RNA or proviral DNA has improved sensitivity in detecting transmitted genotypic drug resistance mutations paired viral RNA and proviral DNA (which is directly extracted from stored whole blood) samples were tested from Ethiopian antiretroviral naive HIV-1C infected subjects. METHODS: In the present comparative study the frequency of TDR mutations was assessed in paired samples of viral RNA and proviral DNA (extracted directly from stored whole blood) of HIV-1C infected treatment naïve patients and interpreted using the 2009 WHO drug resistance surveillance mutation lists, Stanford University drug resistance data base and International Antiviral Society-USA mutation lists. RESULTS: High agreement in rate of TDR between the two compartments was observed using the WHO mutation lists. While mutations G190A and E138A were concurrently found in both compartments, others such as G73S on PR and A62V, M184I, M230I on RT were identified in proviral DNA only. All signature mutations seen in viral RNA were not missed in proviral DNA. CONCLUSIONS: The concordance of major genotype drug resistance mutation between RNA and proviral DNA in treatment naïve patients suggests that proviral DNA might be an alternative approaches for an initial assessment of drug resistance prior to initiation of antiretroviral therapy using the WHO mutations lists in resource-limited countries. However, the clinical importance of TDRMs observed only in proviral DNA in terms of being a risk factor for virologic failure and whether they limit future treatment options needs additional investigation using more sensitive sequencing approaches such as Next Generation Sequencing (NGS).

Methicillin resistant Staphylococcus aureus in Ethiopia: a meta-analysis.

BACKGROUND: The burden of methicillin resistant Staphylococcus aureus is a major public health concern worldwide; however the overall epidemiology of multidrug resistant strains is neither coordinated nor harmonized, particularly in developing countries including Ethiopia. Therefore, the aim of this meta-analysis was to assess the burden of methicillin resistant Staphylococcos aureus and its antibiotic resistance pattern in Ethiopia at large. METHODS: PubMed, Google Scholar, and lancet databases were searched and a total of 20 studies have been selected for meta-analysis. Six authors have independently extracts data on the prevalence of methicillin resistant Staphylococcus aureus among clinical isolates of Staphylococcus aureus. Statistical analysis was achieved by using Open meta-analyst (version 3.13) and Comprehensive meta-analysis (version 3.3) softwares. The overall prevalence of methicillin resistant Staphylococcus aureus and its antibiotic resistance pattern were pooled by using the forest plot, table and figure with 95% CI. RESULTS: The pooled prevalence of methicillin resistant Staphylococcus aureus was 32.5% (95% CI, 24.1 to 40.9%). Moreover, methicillin resistant Staphylococcus aureus strains were found to be highly resistant to penicillin, ampicillin, erythromycin, and amoxicillin, with a pooled resistance ratio of 99.1, 98.1, 97.2 and 97.1%, respectively. On the other hand, comparably low levels of resistance ratio were noted to vancomycin, 5.3%. CONCLUSION: The overall burden of methicillin resistant Staphylococcus aureus is considerably high, besides these strains showed extreme resistance to penicillin, ampicillin, erythromycin and amoxicillin. In principle, appropriate use of antibiotics, applying safety precautions are the key to reduce the spread of multidrug resistant strains, methicillin resistant Staphylococcus aureus in particular.

Cockroaches as a Source of High Bacterial Pathogens with Multidrug Resistant Strains in Gondar Town, Ethiopia.

Background. Cockroaches are source of bacterial infections and this study was aimed to assess bacterial isolates and their antimicrobial profiles from cockroaches in Gondar town, Ethiopia. Methods. A total of 60 cockroaches were collected from March 1 to May 30, 2014, in Gondar town. Bacterial species were isolated from external and internal parts of cockroaches. Disk diffusion method was used to determine antibiotic susceptibility patterns. Data were entered and analyzed by using SPSS version 20; P values <0.005 were considered as statistically significant. Results. Of 181 identified bacteria species, 110 (60.8%) and 71 (39.2%) were identified from external and internal parts of cockroaches, respectively. Klebsiella pneumoniae 32 (17.7%), Escherichia coli 29 (16%), and Citrobacter spp. 27 (15%) were the predominant isolates. High resistance rate was observed to cotrimoxazole, 60 (33.1%), and least resistance rate was noted to ciprofloxacin, 2 (1.1%). Additionally, 116 (64.1%) of the isolates were MDR strains; Salmonella spp. were the leading MDR isolates (100%) followed by Enterobacter (90.5%) and Shigella spp. (76.9%). Conclusion. Cockroaches are the potential source of bacteria pathogens with multidrug resistant strains and hence effective preventive and control measures are required to minimize cockroach related infections.

Limited increase in primary HIV-1C drug resistance mutations in treatment naïve individuals in Ethiopia.

Antiretroviral drug resistance is a major challenge for management and control of HIV-1 infection worldwide and particularly in resource limited countries. The frequency of primary drug resistance mutations (DRMs) and of naturally occurring polymorphisms was determined in 83 antiretroviral treatment (ART) naïve Ethiopian individuals infected with HIV-1, consecutively enrolled in 2010. In all individuals HIV-1C was found. The median (interquartile range) of CD4(+) T-cell count and viral load were 100 (49-201) cells/µl and 44,640 (12,553-134,664) copies/ml, respectively. Protease (PR) and reverse transcriptase (RT) genes of HIV-1 RNA were amplified and sequenced. The proportion of primary DRM to any drug class, using the World Health Organization mutation lists, was 7.2% (6/83), thus exceeding the WHO threshold limit of 5%. Three individuals (3.6%) had non-nucleoside reverse transcriptase inhibitor (NNRTI) mutations, two individuals (2.4%) had protease inhibitor mutations, and one (1.2%) had mutations associated with two drug classes (nucleoside reverse transcriptase inhibitor and NNRTI). In addition, the frequency of polymorphisms in the PR and RT genes was higher compared with previous studies in Ethiopian as well as worldwide isolates. Hence, genotypic drug resistance testing as part of routine management of individuals seems reasonable even in resource limited countries prior to treatment in order to allow proper choice of ART.

The growing challenges of antibacterial drug resistance in Ethiopia.

Infectious diseases of bacterial origin are a major cause of morbidity and mortality in developing countries such as Ethiopia. To minimise such burdens, proper use of antibiotics has played a vital role and saved countless lives. However, use of antimicrobials as therapeutic agents is compromised by the potential development of drug-resistant micro-organisms. Currently, antimicrobial drug resistance has become a public health concern both in developing and developed countries. Antimicrobial drug resistance is dramatically accelerated when antimicrobials are misused. This is critical, especially in developing countries where they are not only misused but are often underused due to financial constraints. Although large-scale studies on antimicrobial resistance in Ethiopia have not yet been conducted, the available reports indicate a trend towards increasing resistance rates among pathogens such as Escherichia coli, Shigella spp., Salmonella spp. and Staphylococcus aureus to commonly prescribed antibiotics, including ampicillin, amoxicillin, penicillin, tetracycline and trimethoprim/sulfamethoxazole. This review summarises the existing data on antibacterial drug resistance in this country.

Intestinal parasitosis and shigellosis among diarrheal patients in Gondar teaching hospital, northwest Ethiopia.

BACKGROUND: Diarrheal diseases are the major causes of morbidity and mortality in developing world. Understanding the etiologic agents of diarrheal diseases and their association with socio-demographic characteristics of patients would help to design better preventive measures. Thus, this study was aimed to determine the prevalence of intestinal parasites and enteropathogenic bacteria in diarrheic patients. METHODS: A cross-sectional study involving 384 consecutive diarrheal patients who visited Gondar teaching hospital, Gondar, Ethiopia from October 2006 to March 2007 was conducted. Stool specimens were collected and examined for intestinal parasites and enteropathogenic bacteria following standard parasitological and microbiological procedures. RESULTS: Intestinal parasites were diagnosed in 36.5% of the patients. The most frequently encountered protozoan parasite was Entamoeba histolytica/dispar (7.3%) followed by Giardia lamblia (5.0%), Cryptosporidium parvum (1.8%) and Isospora belli (1.3%). The dominant helminthic parasite identified was Ascaris lumbricoides (5.5%) followed by Strongyloides stercoralis and Schistosoma mansoni (3.1% each), hookworm infection (1.8%), and Hymenolepis species (1.3%). Multiple infections of intestinal parasites were also observed in 6.3% of the patients. Among the enteropathogenic bacteria Shigella and Salmonella species were isolated from 15.6% and 1.6%, respectively, of the patients. Escherichia coli O57:H7 was not found in any of the stool samples tested. Eighty eight percent and 83.3% of the Shigella and Salmonella isolates were resistant to one or more commonly used antibiotics, respectively.Intestinal parasitosis was higher in patients who live in rural area, in patients who were washing their hands after visiting toilet either irregularly with soap and without soap or not at all, in patients who used well and spring water for household consumption, and in patients who had nausea (P < 0.05). Statistically significant associations were also observed between Shigella infections and patients who were using well and spring water for household consumption, and patients who had dysentery and mucoid stool (P < 0.05). CONCLUSIONS: The high prevalence of intestinal parasites and Shigella species in diarrheic patients calls for institution of appropriate public health intervention measures to reduce morbidity and mortality associated with these diseases. The rational use of antibiotics should also be practiced.

Vitamin A deficiency during pregnancy of HIV infected and non-infected women in tropical settings of Northwest Ethiopia.

BACKGROUND: Vitamin A deficiency (VAD) is known to be a major public health problem among women of reproductive age in South East Asia and Africa. In Ethiopia, there are no studies conducted on serum vitamin A status of HIV-infected pregnant women. Therefore, the present study was aimed at determining the level of serum vitamin A and VAD among pregnant women with and without HIV infection in tropical settings of Northwest Ethiopia. METHODS: In this cross-sectional study, blood samples were collected from 423 pregnant women and from 55 healthy volunteers who visited the University of Gondar Hospital. Serum concentration of vitamin A was measured by high performance liquid chromatography. RESULTS: After controlling for total serum protein, albumin and demographic variables, the mean ± SD serum vitamin A in HIV seropositive pregnant women (0.96 ± 0.42 µmol/L) was significantly lower than that in pregnant women without HIV infection (1.10 ± 0.45 µmol/L, P < 0.05). Likewise, the level of serum vitamin A in HIV seropositive non-pregnant women (0.74 ± 0.39) was significantly lower than that in HIV negative non-pregnant women (1.18 ± 0.59 µmol/L, P < 0.004). VAD (serum retinol < 0.7 µmol/L) was observed in 18.4% and 17.7% of HIV infected and uninfected pregnant women, respectively. Forty six percent of non-pregnant women with HIV infection had VAD while only 28% controls were deficient for vitamin A (P = 0.002). CONCLUSION: The present study shows that VAD is a major public health problem among pregnant women in the tropical settings of Northwest Ethiopia. Considering the possible implications of VAD during pregnancy, we recommend multivitamin (which has a lower level of vitamin A) supplementation in the care and management of pregnant women with or without HIV infection.

Isolation and characterization of Listeria monocytogenes and other Listeria species in foods of animal origin in Addis Ababa, Ethiopia.

Listeriosis is a disease of humans and animals, in which it is one of the important emerging bacterial zoonotic diseases worldwide. Among the different species of the genus Listeria, Listeria monocytogenes (L. monocytogenes) is known to cause listeriosis in humans and animals with low incidence but high case fatality rate. Information on the occurrence and distribution of L. monocytogenes and other Listeria species is very limited both in the veterinary and public health sectors in Ethiopia. The objective of this study was to isolate and characterize L. monocytogenes and other Listeria species from foods of animal origin (cottage cheese, raw beef, raw milk and liquid whole egg) in Addis Ababa, Ethiopia. A total of 391 food samples of animal origin were collected randomly, using a cross-sectional study design from November 2008 to March 2009. L. monocytogenes isolation and characterization were performed according to mainly the United States Food and Drug Administration procedures. Of the samples examined, 102 (26.1%) were found to be positive for Listeria. Listeria species were isolated in 39 (51.3%), 37 (32.2%), 22 (22%) and 4 (4%) of the raw beef, liquid whole egg, raw milk and cottage cheese samples respectively. L. monocytogenes was detected in 5.4% of the samples analyzed. It was isolated mainly from raw milk (13%) and liquid whole egg (4.3%) followed by raw beef (2.6%) and cottage cheese (1%). In addition to L. monocytogenes, other Listeria species were identified as L. innocua (60.8%), L. welshimeri (6.9%), L. seeligeri (3.9%), L. murrayi (2.9%) and L. grayi (2.9%) and L. ivanovii (1.9%). It was shown that L. monocytogenes and other Listeria species are widely spread in occurrence in foods of animal origin in Addis Ababa, Ethiopia.

Immune restoration disease and changes in CD4+ T-cell count in HIV- infected patients during highly active antiretroviral therapy at Zewditu memorial hospital, Addis Ababa, Ethiopia.

BACKGROUND: Highly active antiretroviral therapy (HAART) improves the immune function and decreases morbidity, mortality and opportunistic infections (OIs) in HIV-infected patients. However, since the use of HAART, immune restoration disease (IRD) has been described in association with many OIs. Our objective was to determine the proportion of IRD, changes in CD4+ T-cell count and possible risk factors of IRD in HIV-infected patients. METHODS: A retrospective study of all HIV- infected patients starting HAART between September 1, 2005 and August 31, 2006 at Zewditu memorial hospital HIV clinic, Addis Ababa, Ethiopia was conducted. All laboratory and clinical data were extracted from computerized clinic records and patient charts. RESULTS: A total of 1166 HIV- infected patients with mean ± SD age of 36 ± 9.3 years were on HAART. IRD was identified in 170 (14.6%) patients. OIs diagnosed in the IRD patients were tuberculosis (66.5%, 113/170), toxoplasmosis (12.9%, 22/170), herpes zoster rash (12.9%, 22/170), Pneumocystis jirovecii pneumonia (4.1%, 7/170), and cryptococcosis (3.5%, 6/170). Of the 170 patients with IRD, 124 (72.9%) patients developed IRD within the first 3 months of HAART initiation. Low baseline CD4+ T-cell count (odds ratio [OR], 3.16, 95% confidence interval [CI], 2.19-4.58) and baseline extra pulmonary tuberculosis (OR, 7.7, 95% CI, 3.36-17.65) were associated with development of IRD. Twenty nine (17.1%) of the IRD patients needed to use systemic anti-inflammatory treatment where as 19(11.2%) patients required hospitalization associated to the IRD occurrence. There was a total of 8 (4.7%) deaths attributable to IRD. CONCLUSIONS: The proportion and risk factors of IRD and the pattern of OIs mirrored reports from other countries. Close monitoring of patients during the first three months of HAART initiation is important to minimize clinical deterioration related to IRD.

Assesment of household burden of orphaning and coping strategies by guardians and families with orphans and vulnerable children in Hossana Town, SNNPR.

BACKGROUND: Globally the number of orphans is increasing due to HIV/AIDS, internal corflicts and others. Orphanage and vulnerability causes much of its burden on poor households and children. OBJECTIVE: This study was initiated to assess household challenges of orphaning and coping strategies by families and guardians living with Orphan and Vulnerable Children (OVC). It has also tried to look at the types and level of challenge facing OVC. METHODOLOGY: Community based descriptive cross-sectional quantitative survey was conducted from October to November 2008 using a pre-tested Amharic questionnaire among guardians of OVC in Hossana town. Hadiya zone, SNNPR. A total of 334 samples were selected using a simple random sampling technique. RESULT: Sixty nine percent (69%) of guardians were women. Almost all of the OVC had participated in one domestic activity and 26.9% of OVC were involved in domestic work in other houses. A little more than twenty two percent (22.2%) have history of involvement in productive child labor; of whom 79.7% were found working at the time of data collection Above thirty five percent (35.6%) of respondents believe that the OVC are discriminated and 23.5% of the discriminators were step parents. As household coping mechanism 24.9% reported selling domestic animals, 15.6% selling household equipments and 12.9% sell plots of land which all occurs due to lack of resources in households. CONCLUSION AND RECOMMENDATION: The study concluded that caregivers lack sufficient resources to provide basic needs to OVC. Hence, community based training on micro finance assistance, discrimination and psychological support should be given to guardians and the community in general.

Malnutrition and intestinal parasitic infections in school children of Gondar, North West Ethiopia.

BACKGROUND: In developing countries, malnutrition is a considerable health problem with prevalence ranges of 4-46%, with 1-10% severely malnourished. OBJECTIVES: This study aimed to determine the prevalence of malnutrition and intestinal parasitoses and identify risk factors of malnutrition in schoolchildren. SUBJECTS AND METHODS: A cross-sectional study was carried out on 322 schoolchildren, of age 6 to 14 years, attending private and government primary schools, in Gonder town, North West Ethiopia. The study was conducted from December 2006 to February 2007. Nutritional status of these children was determined using anthropometric parameters (weight-for-age, height-for-age and weight-for-height). Epi Info 2000 software was used to evaluate anthropometric results of each individual and formol-ether concentration technique was employed to identify parasites. RESULTS: The prevalence of underweight, stunting, wasting and intestinal parasitoses was 34.8%, 27%, 50% and 55.6%, respectively. Parasites encountered during the study were Ascaris lumbricoides (17.8%), Trichuiris trichiura (3.4%), hookworm (4.3%), Giardia lamblia (9%), Entamoeba histolytica (2.1%), Schistosoma mansoni (2.4%), Hymenolepis nana (4.7%) and Enterobius vermicularis (0.31%), respectively, in single infections. Only two cases of Strongyloides stercoralis was found in multiple infections and none in single infections. The prevalence of multiple parasitoses was 10.9%. Maternal literacy status, sex and age of the child were significantly associated with malnutrition (p < 0.05). CONCLUSION: Even though chronic type of malnutrition (stunting) was decreasing, indicators of acute type of malnutrition were increasing at a higher rate in the area. Implementation of maternal education and school feeding programs, avoidance of students from getting in contact with waste and river water were recommended to avoid malnutrition and intestinal parasitoses in children of the study area.

Physicochemical and bacteriological quality of bottled drinking water in three sites of Amhara Regional State, Ethiopia.

BACKGROUND: The consumption of bottled drinking water is becoming increasing in Ethiopia. As a result there has been a growing concern about the chemical, physical and bacteriological quality of this product. Studies on the chemical, physical and bacteriological quality of bottled water is quite scarce in Ethiopia. OBJECTIVE: This study was therefore aimed to assess the physicochemical and bacteriological qualities of three factories of bottled drinking water products produced in Amhara region. METHODS: A Laboratory based comparative study was conducted to evaluate the physicochemical and bacteriological quality of three factories of bottled drinking water produced in Amhara region. Analysis on the quality of bottled drinking water from the sources, wholesalers and retailers were made with World Health Organization and Quality and Standards Authority of Ethiopia recommendations. Triplicate samples from three types of bottled drinking water were randomly collected and analyzed from June, 2006 to December, 2006. RESULT: A total of 108 commercial bottled drinking water samples were analyzed. The result showed that except pH of factory A all the physicochemical parameters analyzed were with in the recommended limits. The pH value of factory A tested from sources is 5.3 and from wholesalers and retailers is 5.5 and 5.3, respectively, which is below the normal value set by World Health Organization (6.5-8.0) and Quality and Standards Authority of Ethiopia (6.0-8.5). Our analyses also demonstrated that 2 (16.7%) of the samples tested from sources and 1 (8.3%) from wholesalers of factory B were contaminated with total coliforms, where as 2 (16.7%) samples from retailers were also contaminated with total coliforms. On the other hand, 1 (8.3%) of the samples tested from wholesalers and 2 (16.7%) of the samples tested from retailers of factory A were also contaminated with total coliforms. Total coliforms were not detected from all samples of factory C, fecal coliforms were not also isolated from all samples. Percent of coefficient of variation showed that variations in total coliforms counts were significant with in the samples of both factory A and B (CV > 10%). CONCLUSIONS: Based on the recommended limit of World Health Organization and Quality and Standards Authority of Ethiopia, 7.4% of bottled drinking water sold commercially could be considered unfit for human consumption. Consumers of bottled water should be aware of this.

Prevalence of bacteria and intestinal parasites among food-handlers in Gondar town, northwest Ethiopia.

Food-handlers with poor personal hygiene working in food-service establishments could be potential sources of infection due to pathogenic organisms. The study was undertaken to determine the prevalence of bacteria and intestinal parasites among 127 food-handlers working in the cafeterias of the University of Gondar and the Gondar Teachers Training College, Gondar, Ethiopia. Fingernail contents of both the hands and stool specimens were collected from all the 127 food-handlers. The samples were examined for bacteria and intestinal parasites following standard procedures. Coagulase-negative staphylococci were the predominant bacteria species (41.7%) isolated from fingernail contents, followed by Staphylococcus aureus (16.5%), Klebsiella species (5.5%), Escherichia coli (3.1%), Serratia species (1.58%), Citrobacter species (0.8%), and Enterobacter species (0.8%). Shigella species were isolated from stool samples of four food-handlers (3.1%). None of the food-handlers was positive for Salmonella species and Shigella species in respect of their fingernail contents. No intestinal parasites were detected from fingernail contents. Intestinal parasites detected in the stools of the food-handlers included Ascaris lumbricoides (18.11%), Strongyloides stercoralis (5.5%), Entamoeba histolytica/dispar (1.6%), Trichuris trichiura (1.6%), hookworm species (0.8%), Gardia lamblia (0.8%), and Schistosoma mansoni (0.8%); 1.6% of the study subjects were positive for each of A. lumbricoides, T. trichiura, hookworm, and G. lamblia. The findings emphasize the importance of food-handlers as potential sources of infections and suggest health institutions for appropriate hygienic and sanitary control measures.

Immune reconstitution inflammatory syndrome among HIV/AIDS patients during highly active antiretroviral therapy in Addis Ababa, Ethiopia.

Suppression of viral replication is followed by increases in CD4+ lymphocytes, and this has been shown to result in decreased susceptibility to opportunists after initiation of highly active antiretroviral therapy (HAART). However, clinical aggravations after the initiation of HAART have been thought to be due to the restored ability to mount an inflammatory response, or the immune reconstitution inflammatory syndrome (IRIS). The degree of IRIS observed in human immunodeficiency virus (HIV)-infected patients following initiation of HAART is variable. This prospective study was aimed at determining the proportion of IRIS and the pattern of opportunistic infections among 186 HIV/AIDS patients receiving HAART between December 2006 and July 2007 at Zewditu Memorial Hospital, Addis Ababa, Ethiopia. The proportion of IRIS was 17.2% (32/186). The mean number of days of IRIS occurrence for each disease ranged from 26 to 122 days with a mean of 80. Opportunistic diseases associated with IRIS were tuberculosis (68.8%, 22/32), herpes zoster rash (12.5%, 4/32), cryptococcosis (9.4%, 3/32), toxoplasmosis (6.3%, 2/32) and bacterial pneumonia (3.1%, 1/32). Compared to baseline readings there were significant increases in CD4 count, aspartate aminotransferase and alanine aminotransferase levels while hemoglobin values decreased during the development of IRIS. In summary, the proportion of IRIS and the pattern of opportunistic infections in HAART-treated patients in Ethiopia mirrored those reported in other countries. Further prospective surveys on epidemiological, immunological, microbial and clinical studies are imperative to assess the proportion and pattern of IRIS and effect of HAART in Ethiopia.