Spliced-Leader RNA as a Dynamic Marker for Monitoring Viable Leishmania Parasites During and After Treatment.

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.

Adapting international clinical trials during COVID-19 and beyond.

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.

Vertical Transmission, Risk Factors, and Antimicrobial Resistance Patterns of Group B Streptococcus among Mothers and Their Neonates in Southern Ethiopia.

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.

Bacteriological quality and safety of bottle food and associated factors among bottle-fed babies attending pediatric outpatient clinics of Government Health Institutions in Arba Minch, southern Ethiopia.

BACKGROUND: Microbial contamination of baby bottle food has been identified as a significant public health concern, especially in developing countries, but it remains overlooked. Therefore, this study aimed to determine microbiological hazards, compliance with hygiene practices, and critical control points of contamination in baby bottle food in Arba Minch, southern Ethiopia. OBJECTIVE: To evaluate the bacteriological quality and prevalence of foodborne pathogens in baby bottle food and to identify associated factors among bottle-fed babies attending three government health institutions in Arba Minch, southern Ethiopia. METHODS: A cross-sectional study was conducted between February 24 and March 30, 2022. A total of 220 food samples, comprising four types prepared with different sources of materials, were collected from systematically selected bottle-fed babies attending health facilities. The data on sociodemographic characteristics, food hygiene, and handling practices were solicited by face-to-face interview using a semi-structured questionnaire. Food samples (10 mL) were quantitatively analyzed for total viable counts (TVC) and total coliform count (TCC) and qualitatively for the presence of common foodborne bacterial pathogens. Data were analyzed using SPSS; ANOVA and multiple linear regression analyses were done to identify factors influencing microbial counts. RESULTS: Results revealed that the means and standard deviations of TVC and TCC were 5.3 ± 2.3 log10 colony forming units (CFU)/mL and 4.1 ± 2.6 log10 CFU/mL, respectively. Of the various food samples analyzed, 57.3 and 60.5% had a TVC and TCC above the maximum acceptable limits, respectively. The result of the ANOVA showed that there was a significant difference in the mean score of TCV and TCC among the four types of food samples (p < 0.001). Enterobacteriaceae were found in the majority of positive food samples (79.13%), followed by Gram-positive cocci (20.8%). Salmonella spp., diarrheagenic Escherichia coli, and Staphylococcus aureus were the common foodborne pathogens detected in 8.6% of tested foods. The regression result revealed that the type of baby food, hand washing practices of mothers or caregivers, and sterilizing and disinfecting procedures of feeding bottles are independent determinants of bacterial contamination (p < 0.001). CONCLUSION: The high microbial load and the presence of potential foodborne bacterial pathogens in the bottle food samples analyzed indicate unsanitary practices and the potential risk of exposure to foodborne pathogens in bottle-fed babies. Thus, interventions such as educating parents about proper hygiene practices, sterilizing feeding bottles and limiting bottle feeding practices are critical to reducing the risk of foodborne to bottle-fed infants.

Magnitude and associated factors of bacterial urinary tract infections among paediatric patients in Arba Minch, southern Ethiopia.

Bacterial urinary tract infections (UTI) commonly occur in children; if left untreated, they may result in severe consequences such as uro-sepsis and renal damage. This study aimed to determine the bacterial profile, antimicrobial susceptibility patterns and associated factors among paediatric patients suspected of urinary tract infections in Arba Minch General Hospital (AMGH). An institution-based cross-sectional study was conducted from 01 October 2020 to 31 January 2021. A convenient sampling technique was used to recruit the participants; data were collected using a pre-tested questionnaire. To quantify the bacteria (as per the Kass count, >105CFU/ml), midstream urine samples were streaked onto bacteriological media. Isolates were identified by following standard procedures. The antibiotic susceptibility test was performed as per the Kirby-Bauer disc diffusion technique. Data were analyzed using SPSS software. Out of the 246 children included, 38 (15.4%) were found to be positive for significant bacteriuria. Isolates of Escherichia coli, 9/38 (23.7%), and Staphylococcus aureus, 9/38 (23.7%), were the most predominant. The majority of Gram-negative bacterial (GNB) isolates showed resistance towards amoxicillin-clavulanate (89.5%), ampicillin (84.6%), and ceftazidime (81%). Likewise, 76.9 and 76.5% of Gram-positive bacteria (GPB), respectively, had shown resistance towards co-trimoxazole and tetracycline. Multi-drug and extensively drug resistance were detected respectively in the case of 68.4 and 15.8% of the total isolates; ESBL production was found in 57.1% of GNB, whereas 55.6% of S. aureus were methicillin-resistant S. aureus (MRSA). The process of un-circumcision was significantly associated with UTI [(adjusted odds ratio= 3.578; 95% confidence interval: 1.263 - 10.13; p=0.016)].

Prevalence, severity and associated risk factors of anemia among human immunodeficiency virus-infected adults in Sawla General Hospital, Southern Ethiopia: A facility-based cross-sectional study.

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.

Community-based treatment of cutaneous leishmaniasis using cryotherapy and miltefosine in Southwest Ethiopia: the way forward?

Background: Cutaneous leishmaniasis (CL) is a common, yet massively underreported skin morbidity in Ethiopia. Most patients never seek treatment, as this is offered only in specialized treatment centers. Early diagnosis and treatment through decentralization is crucial to decrease transmission and to reach the NTD roadmap goals. However, little information is available on outcomes and challenges of community-based treatment initiatives. Methods: A community-based prospective cohort study was conducted in Ochollo. Patients with clinically or microscopy confirmed CL were included. Cryotherapy was (to be) given weekly with at least four sessions for uncomplicated lesions, and miltefosine was given for 4 weeks for complicated lesions. Miltefosine adherence was assessed by counting pill strips. Clinical and patient-reported outcomes (dermatological life quality index and patient-global assessment) were assessed at month 6 (M6). Results: A total of 107 patients were included, with a median age of 6 years. Two patients refused, and 15 could not be treated as they were too young (<4 years) for miltefosine. Giving cryotherapy to patients weekly was not feasible due to long wound healing times and required use of topical antibiotics. Only 52.4% of miltefosine patients finished >90% of their tablets by M1. Among 46 patients treated with cryotherapy, 24 (52.2%) were cured at M6, and 9 (19.6%) had substantial improvement. The cure rate was 16/39 (41.0%) for miltefosine with 28.2% (11/39) substantial improvement. Before treatment, more than half (57.8%) of patients reported that CL did not negatively impact their life, which significantly increased to 95.2% at M6. At this time, 61.7% of patients said their lesion was clear, which was 1% before treatment. Conclusion: Our study is the first to identify the challenges and opportunities of miltefosine and cryotherapy for community treatment of CL. Although overall cure rates were lower than expected, patient-reported outcomes were generally positive and quite some patients had good improvement.

First report of cutaneous leishmaniasis caused by Leishmania donovani in Ethiopia.

BACKGROUND: Leishmaniasis is a common neglected tropical disease in Ethiopia. Visceral leishmaniasis (VL) caused by Leishmania donovani presents in the lowlands, while cutaneous leishmaniasis (CL) affects people living in the highlands. Although CL is described as being caused by Leishmania aethiopica, there is also evidence of L. tropica and L. major isolated from a patient, sand flies and potential reservoirs. Information on species causing CL in Ethiopia is patchy, and no nation-wide study has ever been done. Understanding which species are causing CL in Ethiopia can have important implications for patient management and disease prevention. METHODS: We analyzed stored routine samples and biobanked DNA isolates from previously conducted studies of CL patients from different centers in the north, center and south of Ethiopia. Species typing was performed using ITS-1 PCR with high-resolution melt (HRM) analysis, followed by HSP70 amplicon sequencing on a selection of the samples. Additionally, sociodemographic, clinical and laboratory data of patients were analyzed. RESULTS: Of the 226 CL samples collected, the Leishmania species could be determined for 105 (45.5%). Leishmania aethiopica was identified in 101 (96.2%) samples from across the country. In four samples originating from Amhara region, northwestern Ethiopia, L. donovani was identified by ITS-1 HRM PCR, of which two were confirmed with HSP70 sequences. While none of these four patients had symptoms of VL, two originated from known VL endemic areas. CONCLUSIONS: The majority of CL was caused by L. aethiopica, but CL due to L. tropica and L. major cannot be ruled out. Our study is the first to our knowledge to demonstrate CL patients caused by L. donovani in Ethiopia. This should spark future research to investigate where, how and to which extent such transmission takes place, how it differs genetically from L. donovani causing VL and whether such patients can be diagnosed and treated successfully with the currently available tools and drugs.

Indoor air microbial load, antibiotic susceptibility profiles of bacteria, and associated factors in different wards of Arba Minch General Hospital, southern Ethiopia.

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.

Buruli Ulcer and Medical Geo-Microbiology.

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.

Magnitude of Malaria-Typhoid Fever Coinfection in Febrile Patients at Arba Minch General Hospital in Southern Ethiopia.

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.

Bacteriological Profiles, Antimicrobial Susceptibility Patterns, and Associated Factors in Patients Undergoing Orthopedic Surgery with Suspicion of Surgical Site Infection at Arba Minch General Hospital in Southern Ethiopia.

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.

Variation in Glucose-6-Phosphate Dehydrogenase activity following acute malaria.

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.

In Vitro Antagonistic Effect of Lactic Acid Bacteria Isolated from Fermented Beverage and Finfish on Pathogenic and Foodborne Pathogenic Microorganism in Ethiopia.

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.

Antibody and cytokine levels in visceral leishmaniasis patients with varied parasitemia before, during, and after treatment in patients admitted to Arba Minch General Hospital, southern Ethiopia.

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-ß1 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-ß1 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.

Detection of Cutaneous Leishmaniasis Foci in South Ethiopia.

Cutaneous leishmaniasis (CL) is a major public health problem in Ethiopia. The disease is endemic in Ochollo, a village in southern Ethiopia, but there are no reports of CL in the wider area, although it is ecologically very similar. We conducted a rapid assessment survey in the South Ethiopian Rift Valley and found 100 parasitologically confirmed CL cases in 38 villages not reported endemic for CL. Approximately half of the cases were children (57%), and most lesions occurred on the face (78%) and were older than 6 months (77%). Only 2% of the people was aware of the mode of transmission, and 9% sought modern treatment at a hospital. These preliminary data indicate that CL is much more widespread than previously reported and that the disease might have a large psychosocial impact. Hence, this study calls for larger surveys across the Ethiopian highlands. Additionally, health education and treatment capacity need to be implemented.

Evaluation of conventional and four real-time PCR methods for the detection of Leishmania on field-collected samples in Ethiopia.

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.

Epidemiology of visceral leishmaniasis in Shebelle Zone of Somali Region, eastern Ethiopia.

BACKGROUND: Visceral leishmaniasis (VL), a vector-borne disease caused by species of the L.donovani complex, has (re)-emerged in Ethiopia during the last two decades and is currently of increasing public health concern. However, very little is known about VL epidemiology in the Somali Region of Ethiopia. The aim of this study was to provide detailed epidemiological information on seroprevalence, associated factors and incriminated vectors of VL in Shebelle Zone and Ethiopian Somali Region in general. METHODS: A cross-sectional epidemiological study was conducted between March and May 2016 in Gode and Adadle districts of Shebelle Zone, Ethiopian Somali Region. Two-stage semi-random sampling was applied for selecting study participants for the field survey. The study included structured questionnaire interviews, serological assays (rK39-immunochromatographic test), ELISA and entomological surveys. RESULTS: From a total of 361 participants, 57 (15.8%) were seropositive for VL including 46 (12.7%) rK39 positive and 11 (3.0%) positive by both rK39 and ELISA. VL seroprevalence was higher (P < 0.001) in Adadle (31.1%) compared to Gode (12.7%) district. The VL seroprevalence rate was higher in females than in males [rK39 (17.2 vs 14.0%) and ELISA (3.4 vs 2.5%)]. Children under the 15 years of age were the most highly affected group [rK39 (20.4%) and ELISA (4.4%)]. Increased VL risk was associated with presence of termite hills, study district, outdoor sleeping, Acacia trees and domestic animals [odds ratio (95% confidence interval): 12.58 (5.911-26.763), 5.40 (2.90-10.07), 5.31 (2.283-12.364), 2.37 (1.1190-4.728) and 0.199 (0.097-0.410), respectively]. The entomological survey identified 74 Phlebotomus [P. (Larroussius) orientalis (52/74), P. (Anaphlebotomus) rodhaini (14/74), P. (Paraphlebotomus) sergenti (8/74)] and 11 Sergentomyia sand flies. The average frequency of P. orientalis (3.06 ± 0.66) collected by all traps per night was higher than that of other species. The average frequency of total and specific (P. orientalis) female sand flies was higher in Adadle (1.89 ± 0.423 vs 1.11 ± 0.309) than in Gode (0.62 ± 0.324 vs 0.38 ± 0.183) district. The highest mean numbers of total (8 ± 1.5) and P. orientalis (6 ± 0.913) sand flies were collected in termite hills. CONCLUSIONS: The present findings revealed potential new VL-transmission foci in the study districts. Therefore, the need for parasitological and molecular characterization of the parasite in humans and vector sand flies is of paramount importance to confirm transmission.

Polymorphism in the HASPB repeat region of East African Leishmania donovani strains.

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.