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Accurate detection of viable Leishmania parasites is critical for evaluating visceral leishmaniasis (VL) treatment response at an early timepoint. We compared the decay of kinetoplast DNA (kDNA) and spliced-leader RNA (SL-RNA) in vitro, in vivo, and in a VL patient cohort. An optimized combination of blood preservation and nucleic acid extraction improved efficiency for both targets. SL-RNA degraded more rapidly during treatment than kDNA, and correlated better with microscopic examination. SL-RNA quantitative polymerase chain reaction emerges as a superior method for dynamic monitoring of viable Leishmania parasites. It enables individualized treatment monitoring for improved prognoses and has potential as an early surrogate endpoint in clinical trials. [ABSTRACT FROM AUTHOR]

Background: Anemia is a significant public health problem in HIV/AIDS patients worldwide. This study is aimed to determine the prevalence of anemia and its risk factors among HIV-infected adults in Sawla General Hospital, southern Ethiopia. Methods: A facility-based cross-sectional study involving HIV-infected adults was conducted in ART clinic of Sawla General Hospital from April 01 to May 31, 2019. A systematic random sampling technique was employed to recruit the study participants. Socio-demographic and clinical data were collected using a structured questionnaire and checklist. Hemoglobin concentration from venous blood was determined by HemoCue® 301 analyzer. Descriptive and inferential statistics, by Statistical Package for Social Science version 26.0, were applied; p-values ≤ 0.05 in the multivariable logistic regression analysis were considered statistically significant. Results: A total of 220 HIV-infected adults participated in this study. The prevalence of anemia was 38.6%, from which 90.6, 7.1, and 2.3% are mild, moderate, and severe anemia, respectively. Anemia among HIV-infected adults was significantly associated with CD4 cell count below 200 cells/mm3 (AOR: 4.32; 95% CI: 2.10–8.86), clinical stage III or above (AOR: 4.20; 95% CI: 1.06–16.62), five or more years duration of HIV infection (AOR: 2.32; 95% CI: 1.08–4.94) and BMI below 18.5 kg/m2 (AOR: 3.82; 95% CI: 1.83–8.00). Conclusion: Anemia is a moderate public health problem among the study population. Longer duration of HIV infection, advanced clinical stage, lower CD4 cell count, and BMI are risk factors for anemia. Therefore, early ART enrolment for HIV-infected adults with nutritional support and rigorous monitoring of CD4 cell count are essential to lower the prevalence. [ABSTRACT FROM AUTHOR]

Background: The COVID-19 pandemic and resulting restrictions, particularly travel restrictions, have had significant impact on the conduct of global clinical trials. Our clinical trials programme, which relied on in-person visits for training, monitoring and capacity building across nine low- and middle-income countries, had to adapt to those unprecedented operational challenges. We report the adaptation of our working model with a focus on the operational areas of training, monitoring and cross-site collaboration. The new working model: Adaptations include changing training strategies from in-person site visits with three or four team members to a multi-pronged virtual approach, with generic online training for good clinical practice, the development of a library of study-specific training videos, and interactive virtual training sessions, including practical laboratory-focused training sessions. We also report changes from in-person monitoring to remote monitoring as well as the development of a more localized network of clinical trial monitors to support hybrid models with in-person and remote monitoring depending on identified risks at each site. We established a virtual network across different trial and study sites with the objective to further build capacity for good clinical practice–compliant antimalarial trials and foster cross-country and cross-study site collaboration. Conclusion: The forced adaptation of these new strategies has come with advantages that we did not envisage initially. This includes improved, more frequent engagement through the established network with opportunities for increased south-to-south support and a substantially reduced carbon footprint and budget savings. Our new approach is challenging for study sites with limited prior experience but this can be overcome with hybrid models. Capacity building for laboratory-based work remains difficult using a virtual environment. The changes to our working model are likely to last, even after the end of the pandemic, providing a more sustainable and equitable approach to our research. [ABSTRACT FROM AUTHOR]

Buruli ulcer is a chronic debilitating infectious disease caused by the pathogen Mycobacterium ulcerans, which can be cured if diagnosed and treated in an early stage. However, advanced cases need antibiotic treatment followed by surgical interventions. In this context, an extremely effective and less expensive treatment modality can be developed by means of an extended topical application of certain selected natural clay minerals, most of the time containing illite-smectite having some iron content. There is a scope for developing the speciality, medical geo-microbiology, which is truly a multidisciplinary one, for finding a cure for the severe and advanced cases of BU. [ABSTRACT FROM AUTHOR]

Background. Coinfection with malaria and typhoid fever is a major public health issue in developing countries. In endemic areas, including Ethiopia, people are at risk of acquiring both malaria and typhoid fever at the same time. Therefore, this study aimed to determine the magnitude of malaria-typhoid fever coinfection in febrile patients attending hospital at Southern Ethiopia. Methods. A hospital-based cross-sectional study was carried out on 416 febrile patients attending Arba Minch General Hospital from 1st October to 30th December 2021. The data was collected using a pretested structured questionnaire. Capillary and Venus blood samples were collected for assessing malaria and typhoid fever, respectively. Blood smear, culture, and biochemical tests were performed based on standard parasitological and microbiological methods. The P -value ≤ 0.05 was considered statistically significant. Results. The magnitude of malaria, typhoid fever, and their coinfections was 26.2% (109/416), 6.5% (27/416), and 3.1% (13/416), respectively. Among the confirmed malaria cases, about 66% of infections were Plasmodium falciparum. The malaria-typhoid fever coinfection showed a statistically significant association with a clinical presentation of a continuous pattern of fever (AOR = 5.84; 95% CI: 1.44–23.71, P = 0.014) and chills (AOR = 3.94; 95% CI: 1.04–14.89, P = 0.044). About 29.6% of Salmonella isolates were multidrug-resistant (MDR). Conclusion. The total rate of coinfection with malaria and typhoid fever was comparable to that of previous studies. With the consideration of higher prevalence of drug resistance of Salmonella spp. and higher prevalence of malaria‐typhoid fever coinfection, proper diagnostic procedure should be implemented for proper use of drugs. [ABSTRACT FROM AUTHOR]

Background. Group B Streptococcus (GBS) contributes to maternal and neonatal morbidity and mortality by increasing intrauterine infection or vertical transmission at the time of birth. Despite many efforts to reduce the potential risk of vertical transmission, GBS remains the main cause of serious disease (neonatal sepsis, meningitis, and/or pneumonia) in vulnerable newborns during the first week of life. This study aimed to assess vertical transmission, risk factors, and antimicrobial resistance patterns of GBS among pregnant women and their neonates. Methods. A facility-based cross-sectional study was conducted among mothers and their neonates from February to May 2021. A total of 201 pregnant women with their neonates participated in this study. A well-designed questionnaire was used to collect sociodemographic and clinical data. A vaginal swab from mother before delivery and neonatal nasal and ear canal swab samples were taken as soon as after delivery within 30 minutes. Vaginal swabs, neonatal ear canal, and nasal swabs were placed into Todd–Hewitt broth and incubated at 37°C for 18–24 hours at 35–37°C in 5% CO2 conditions and then subcultured on 5% sheep blood agar for 18–48 hours. Presumptive identification of GBS was made by morphology, Gram stain, catalase, and hemolytic activity on sheep blood agar plates. CAMP and bacitracin susceptibility tests were used as confirmatory tests for GBS. Data were analyzed using SPSS version 21. P value ≤0.05 was considered statistically significant. Results. Vertical transmission rates of GBS (mother to neonates) were 11.9%. The prevalence of GBS among pregnant women and newborns was 24/201 (11.9%) (95% CI = 7.5–16.9) and 11/201 (5.5%) (95% CI = 2.5–9.0), respectively. The history of prolonged rupture of membranes (AOR = 3.5, CI = 2.2–18.8) and urinary tract infection (AOR = 2.9, CI = 1.7–16.3) were associated factors for maternal GBS colonization. Gestational age of <37 weeks (p = 0.008), low birth weight of <2.5 kg (p = 0.001), and maternal history of vaginal discharge (p = 0.048) were associated factors for neonatal GBS colonization. Low antibiotic resistance was observed for erythromycin 8.6%, clindamycin 5.7%, and chloramphenicol 2.9%. Conclusion. In this study, high vertical transmission (mother to neonates) rate was observed. The prevalence of vaginal GBS colonization of women at delivery was 11.9% and significantly associated with the history of prolonged rupture of membranes and urinary tract infections. Gestational age of <37 weeks, low birth weight of <2.5 kg, and maternal history of vaginal discharge were associated with neonatal GBS colonization. Hence, there is a need for antenatal culture-based GBS screening, risk factor-based interventions, and regular follow-up of drug resistance patterns for proper treatment and management of GBS. [ABSTRACT FROM AUTHOR]

The levels of indoor air microbial load in hospitals are very crucial to the health of patients and health care workers and are to be regularly monitored and maintained at an acceptable level. However, this problem remains overlooked, particularly in developing countries including Ethiopia. A hospital-based cross-sectional study is designed to determine the indoor air microbial load (settle plate technique), microbial isolates (standard microbiological techniques), bacterial susceptibility profiles (Kirby-Bauer disk diffusion technique), and associated factors, in different wards of the title Hospital, southern Ethiopia. An observational checklist was used to collect relevant information related to the associated factors; descriptive and inferential statistics were applied using Statistical Package for Social Sciences (SPSS); p-values ≤ 0.05 in the multivariable analysis were considered statistically significant. The total average bacterial and fungal load of the selected wards was 1914±1081.4 Colony Forming Units (CFU)/m3 (95% CI: 1718.5–2109.48 CFU/m3) and 1533.7±858.8 CFU/m3 (95% CI: 1378.5-1688CFU/m3) respectively. The highest mean bacterial (1914±1081.4 CFU/m3) and fungal (1533.7±858.8 CFU/m3) loads were found in the male surgical and female medical wards respectively. A total of 229 bacterial and 139 fungal isolates were obtained; Gram-positive bacteria were the predominant type, 130 (56.7%), particularly the isolates of Staphylococcus aureus, 46 (20.1%). The predominant fungal isolates were Aspergillus sp., 53(38%). Percentages of multidrug-resistant (MDR), extended-spectrum beta-lactamase (ESBL), and carbapenemase producers respectively were 48.5, 26.5, and 25%. High room crowd index [p = 0.003; Adjusted Odds Ratio (AOR) 12.5 (Confidence Interval (CI) 95%: 2.42–65)], presence of damp/wet materials [p = 0.025; AOR 7 (CI 95%: 1.3–37.4)], intense room traffic [p = 0.004; AOR 9.6 (CI 95%: 1.2–79.3)], inappropriate storage of food and drugs [p = 0.008; AOR 7.5 (CI 95%: 1.7–32)], and unclean environment [p = 0.03; AOR 5.8 (CI 95%: 1.2–28)] showed statistical significance concerning the indoor air microbial loads; most of the wards in Arba Minch General Hospital (AMGH) stand high and not in an acceptable level as per the WHO and the European Commission standards on indoor air microbial load. Periodic air surveillance and infection prevention control programs are required to reduce the transmission of these microbes to inpatients, visitors, and health care workers. [ABSTRACT FROM AUTHOR]

Primaquine and tafenoquine are the only licensed drugs with activity against Plasmodium vivax hypnozoites but cause haemolysis in patients with glucose–6–phosphate dehydrogenase (G6PD) deficiency. Malaria also causes haemolysis, leading to the replacement of older erythrocytes with low G6PD activity by reticulocytes and young erythrocytes with higher activity. Aim of this study was to assess the impact of acute malaria on G6PD activity. Selected patients with uncomplicated malaria were recruited in Bangladesh (n = 87), Indonesia (n = 75), and Ethiopia (n = 173); G6PD activity was measured at the initial presentation with malaria and a median of 176 days later (range 140 to 998) in the absence of malaria. Among selected participants (deficient participants preferentially enrolled in Bangladesh but not at other sites) G6PD activity fell between malaria and follow up by 79.1% (95%CI: 40.4 to 117.8) in 6 participants classified as deficient (<30% activity), 43.7% (95%CI: 34.2 to 53.1) in 39 individuals with intermediate activity (30% to <70%), and by 4.5% (95%CI: 1.4 to 7.6) in 290 G6PD normal (≥70%) participants. In Bangladesh and Indonesia G6PD activity was significantly higher during acute malaria than when the same individuals were retested during follow up (40.9% (95%CI: 33.4–48.1) and 7.4% (95%CI: 0.2 to 14.6) respectively), whereas in Ethiopia G6PD activity was 3.6% (95%CI: -1.0 to -6.1) lower during acute malaria. The change in G6PD activity was apparent in patients presenting with either P. vivax or P. falciparum infection. Overall, 66.7% (4/6) severely deficient participants and 87.2% (34/39) with intermediate deficiency had normal activities when presenting with malaria. These findings suggest that G6PD activity rises significantly and at clinically relevant levels during acute malaria. Prospective case-control studies are warranted to confirm the degree to which the predicted population attributable risks of drug induced haemolysis is lower than would be predicted from cross sectional surveys. Author summary: Plasmodium vivax forms dormant liver stages in the human host that reactivate weeks to months after the first infection and cause significant morbidity and mortality in affected populations. The group of 8-aminoquinolines are the only class of licensed drugs that remove these liver stages from the human host but are contra-indicated in patients with low activities of the glucose-6-phosphate dehydrogenase enzyme (G6PD). The WHO therefore recommends testing G6PD activity prior to treatment to exclude individuals with low activities from standard treatment. We enrolled 335 patients with malaria in Bangladesh, Indonesia, and Ethiopia and measured G6PD activity in all participants. All participants were followed up and a second G6PD measurement was collected between 6 and 33 months after enrolment if participants were free of malaria. When comparing both measurements, G6PD activity was 10% higher during malaria. The increase in G6PD activity during malaria is probably triggered by the Plasmodium infection, the observed change in G6PD activity may alter the risk profile of standard malaria treatment with 8-aminoquinolines. [ABSTRACT FROM AUTHOR]

Background: In the thoughts of all orthopedicians, the emergence of drug-resistant and biofilm-forming bacterial infections at orthopedic surgical sites is the most feared problem. Thus, this study aimed to determine the bacteriological profiles, antimicrobial susceptibility patterns, and biofilm forming ability of isolates, as well as factors associated with orthopedic surgical site infections (OSSIs). Methods: An institution-based cross-sectional study was conducted from March 1st, 2021, to February 30th, 2022 at Arba Minch General Hospital. About 245 suspected orthopedic patients with surgical site infection were enrolled and structured questionnaires were used to collect the required information. Wound swabs or pus aspirates were aseptically collected. The frequency and type of bacterial pathogen(s), antimicrobial susceptibility pattern, and biofilm formation were used to determine and characterize the magnitude of OSSIs. SPSS version 25 was used to analyze factors associated with OSSIs. Results: The overall magnitude of symptomatic OSSIs was 29.4% (72/245). External fixation [AOR = 4.761, 95% CI: (1.108– 20.457)], implant use [AOR = 3.470, 95% CI: (1.460– 8.246)], length of time for surgery [AOR = 3.225, 95% CI: (1.545– 6.731)], and post-operative hospitalization [AOR = 4.099, 95% CI: (2.026– 8.293)] were all statistically significant. Staphylococcus aureus was the most frequently isolated bacteria, accounting for 76%. Methicillin-resistant was observed in 57.9% and 40% of isolated S. aureus (MRSA) and coagulase-negative staphylococci (CoNS), respectively. One-third of the isolated E. faecium was vancomycin-resistant (VRE). Overall, 67.1% (51/76) of isolates were multidrug-resistant (MDR). About 27.6% (21/76) of isolates were found to be strong biofilm producers. Conclusion: OSSIs were shown to be caused by a significant number of drug-resistant and biofilm-producing bacterial isolates. To mitigate the problem, aseptic surgical practice and conventional wound management, as well as constant observation of antimicrobial resistant patterns, should be followed. [ABSTRACT FROM AUTHOR]

Background. Lactic acid bacteria from fermented foods and fish can antagonistically inhibit the growth of foodborne pathogenic organism in fermented food and they stimulate the immune response to protect the fish from certain kinds of infections. The aim of this study was to evaluate the in vitro antagonistic activities of lactic acid bacteria isolated from fermented beverage (Borde) and finfish on foodborne pathogenic microorganisms. Methods. Laboratory-based experimental study was conducted from May 1 to Sep 1, 2020. Total sample numbers were 60 samples of fermented beverage (Borde) and 20 of finfish which were collected from different households and Chamo Lake (Arba Minch, Ethiopia). Each sample was firstly homogenized and serial dilution was prepared and spread on MRS agar plates in order to isolate pure culture. Different biochemical tests were performed to identify isolated bacteria. Then, cell-free supernatant (CFS) was prepared from MRS culture and used in an antimicrobial assay that was performed by agar diffusion method. The effects of pH, temperature, and enzymes on antimicrobial activity were evaluated in the same test. Simultaneously, the effects of lactic acid bacteria on aflatoxin production and on the permeability of the membrane were also evaluated. Data were analyzed using one-way ANOVA and Tukey post hoc analysis was performed by SPSS 25 statistical software. Result. A total of 40 lactic acid bacteria were isolated; among them, 4 lactic acid bacteria, belonging to the genera Enterococcus, Leuconostoc, and Weisellia from fermented beverage and Pediococcus from fish, were screened for antimicrobial activity. The cell-free supernatant of those four isolates exhibited a significant (p < 0.05) antibacterial effect against tested pathogens and foodborne pathogenic bacteria. In addition, CFS showed antifungal and antiaflatoxigenic activities. The antimicrobial compounds synthesized by these isolates were sensitive to some proteolytic enzymes, and they were proved to be stable at high temperatures. It maintained/retained antimicrobial activity in a wide range of pH 2.0–10. Enterococcal CFS exhibited antibacterial activity against S. aureus on membrane permeability, as confirmed by the increase in absorbance value between 0.075 and 0.24 at OD280-nm and between 0.68 and 1.2 at OD260-nm. Conclusion. Cell-free supernatant produced by isolated lactic acid bacteria showed antimicrobial activity against a wide range of Gram-positive and Gram-negative foodborne bacteria, suggesting its potential application as a natural antimicrobial agent in tackling the rising drug resistance against foodborne pathogens. [ABSTRACT FROM AUTHOR]

Background: Visceral leishmaniasis is a disease caused by disseminated Leishmania donovani infection which affects almost half a million people annually. Most of the patients are reported from the Indian sub-continent, Eastern Africa and Brazil. In this study, we aimed to determine the levels of antibodies and cytokines in visceral leishmaniasis patients and to examine associations of parasitemia with the clinical states of patients. A prospective study was carried out, enrolling a total of 48 active VL patients who were evaluated before, during different time points and, three months after treatment. Serum cytokine concentrations, antibody levels, parasitemia, laboratory (hematologic and biochemical) measurements, and clinical parameters were assessed. Results: Counts of WBC and platelets, and measurements of hemoglobin (Hb) increased during treatment (P ≤ 0.05). Elevated levels of circulating IL-10, IFN-γ, and TGF-β1 were measured before treatment. The observed increase in serum IL-10 remarkably declined within 7 days after the start of treatment. Anti-leishmanial antibody index (AI) was high in all VL patients irrespective of spleen aspirate parasite grade before treatment and at different times during treatment. However, a significant (P ≤ 0.05) decrease of AI was observed 120 days post-treatment. IL-2 serum levels were below the detection limit at all sampling points. Conclusions: The present results suggest that IL-10, IFN-γ, and TGF-β1 can be used as markers of active visceral leishmaniasis. In addition, measuring circulating cytokines concentrations, particularly IL-10, in combination with other clinical evaluations, could be used as criteria for the cure. The observation that a high serum concentration of IFN-gamma at baseline was associated with low parasitemia deserves further investigations. Author summary: Visceral leishmaniasis (also known as kala-azar) is a neglected tropical disease that occurs in widely dispersed areas of the world, including Ethiopia. Parasites in the Leishmania donovani complex are responsible for causing visceral leishmaniasis. The condition is difficult to diagnose and treat. We investigated how the immune response generated during follow-up treatment periods of active VL before, during, and post-treatment was influenced by the presence of different cytokines. It is important to identify possible immunological biomarkers that could be correlated with patients' clinical and parasitological presentation as well as the response patterns to treatment in VL patients of southwestern Ethiopia. [ABSTRACT FROM AUTHOR]

In most low-resource settings, microscopy still is the standard method for diagnosis of cutaneous leishmaniasis, despite its limited sensitivity. In Ethiopia, the more sensitive molecular methods are not yet routinely used. This study compared five PCR methods with microscopy on two sample types collected from patients with a suspected lesion to advise on optimal diagnosis of Leishmania aethiopica. Between May and July 2018, skin scrapings (SS) and blood exudate from the lesion spotted on filter paper (dry blood spot, DBS) were collected for PCR from 111 patients of four zones in Southern Ethiopia. DNA and RNA were simultaneously extracted from both sample types. DNA was evaluated by a conventional PCR targeting ITS-1 and three probe-based real-time PCRs: one targeting the SSU 18S rRNA and two targeting the kDNA minicircle sequence (the 'Mary kDNA PCR' and a newly designed 'LC kDNA PCR' for improved L. aethiopica detection). RNAs were tested with a SYBR Green-based RT-PCR targeting spliced leader (SL) RNA. Giemsa-stained SS smears were examined by microscopy. Of the 111 SS, 100 were positive with at least two methods. Sensitivity of microscopy, ITS PCR, SSU PCR, Mary kDNA PCR, LC kDNA PCR and SL RNA PCR were respectively 52%, 22%, 64%, 99%, 100% and 94%. Microscopy-based parasite load correlated well with real-time PCR Ct-values. Despite suboptimal sample storage for RNA detection, the SL RNA PCR resulted in congruent results with low Ct-values. DBS collected from the same lesion showed lower PCR positivity rates compared to SS. The kDNA PCRs showed excellent performance for diagnosis of L. aethiopica on SS. Lower-cost SL RNA detection can be a complementary high-throughput tool. DBS can be used for PCR in case microscopy is negative, the SS sample can be sent to the referral health facility where kDNA PCR method is available. Author summary: Cutaneous leishmaniasis is a neglected tropical disease and causing a public health problem in Ethiopia. Microscopy is still the standard method for detection of the parasite in Ethiopia, and also in many other low resource settings. A more sensitive method is needed for timely diagnosis and treatment. In this study, we compared five molecular methods on samples collected from patients with a skin lesion suspected of cutaneous leishmaniasis to advice on optimal diagnosis of L. aethiopica. We collected two sample types from the same lesion (skin scrapings and lesion fluid on filter paper) and isolated both DNA and RNA of them. Majority (90.1%) of the samples from skin scrapings were positive in two or more methods and the molecular methods had a higher sensitivity than the conventional methods. Interestingly, we evaluated for the first time a new molecular method designed to improve L. aethiopica detection. Also, we showed that RNA detection performed well for samples that were collected under difficult field conditions. Samples collected on filter paper showed less positive results than skin scraped samples, but could still be the method of choice for easy sampling and transport in resource-limited settings as it performed better than microscopy. [ABSTRACT FROM AUTHOR]

Background/Objectives: Visceral leishmaniasis (VL) caused by Leishmania donovani is a major health problem in Ethiopia. Parasites in disparate regions are transmitted by different vectors, and cluster in distinctive genotypes. Recently isolated strains from VL and HIV-VL co-infected patients in north and south Ethiopia were characterized as part of a longitudinal study on VL transmission. Methodology/Principal Findings: Sixty-three L. donovani strains were examined by polymerase chain reaction (PCR) targeting three regions: internal transcribed spacer 1 (ITS1), cysteine protease B (cpb), and HASPB (k26). ITS1- and cpb - PCR identified these strains as L. donovani. Interestingly, the k26 - PCR amplicon size varied depending on the patient's geographic origin. Most strains from northwestern Ethiopia (36/40) produced a 290 bp product with a minority (4/40) giving a 410 bp amplicon. All of the latter strains were isolated from patients with HIV-VL co-infections, while the former group contained both VL and HIV-VL co-infected patients. Almost all the strains (20/23) from southwestern Ethiopia produced a 450 bp amplicon with smaller products (290 or 360 bp) only observed for three strains. Sudanese strains produced amplicons identical (290 bp) to those found in northwestern Ethiopia; while Kenyan strains gave larger PCR products (500 and 650 bp). High-resolution melt (HRM) analysis distinguished the different PCR products. Sequence analysis showed that the k26 repeat region in L. donovani is comprised of polymorphic 13 and 14 amino acid motifs. The 13 amino acid peptide motifs, prevalent in L. donovani, are rare in L. infantum. The number and order of the repeats in L. donovani varies between geographic regions. Conclusions/Significance: HASPB repeat region (k26) shows considerable polymorphism among L. donovani strains from different regions in East Africa. This should be taken into account when designing diagnostic assays and vaccines based on this antigen. Author Summary: HASPB belongs to a hydrophilic repeat-containing surface antigen family found in Leishmania. The L. infantum/L. donovani protein has been used for diagnosis of visceral leishmaniasis, and is a putative vaccine candidate for this disease. Visceral leishmaniasis is a fatal disease, and approximately one third of the cases are found in East Africa. The k26 – PCR, which amplifies the repeat region of HASPB, produced different amplicon sizes for recent Ethiopian L. donovani depending on the strain's geographic origin. Further analysis showed that the number and order of the peptide motifs, either 13 or 14 amino acids long, comprising the L. donovani repeats varies between endemic regions of East Africa. Polymorphism in the amino acid sequence of the peptides was also observed. In addition, the 13 amino acid peptide motifs prevalent in L. donovani are rare in L. infantum. The observed polymorphisms in the HASPB repeat region suggests that custom antigens may be needed for diagnosis or vaccination in distinct endemic foci. [ABSTRACT FROM AUTHOR]