Evaluation of five diagnostic methods for Strongyloides stercoralis infection in Amhara National Regional State, northwest Ethiopia

Background: Strongyloides stercoralis is an intestinal parasite that can cause chronic infection, hyperinfection and/or a dissemination syndrome in humans. The use of techniques targeting ova fails to detect S. stercoralis, as only larvae of the parasite are excreted in faeces. Due to the absence of "Gold" standard diagnostic method for S. stercoralis, there is a paucity of reported data worldwide. Objective: This study aimed to evaluate the performance of diagnostic methods of S. stercoralis infection by taking the composite reference as a "Gold" standard. Methods: A cross-sectional study was conducted among 844 schoolchildren in Amhara Region, Ethiopia, from April to December 2019. Stool samples were collected and processed with formol-ether concentration technique (FECT), spontaneous tube sedimentation technique (STST), Baermann concentration technique (BCT), agar plate culture (APC) and real-time polymerase chain reaction (RT-PCR). Sensitivity, specificity, positive predictive value, and negative predictive value of each diagnostic method were computed against the composite reference. The agreements of diagnostic methods were evaluated by Kappa value at 95% CI. Results: The composite detection rate of S. stercoralis by the five diagnostic methods was 39.0% (329/844). The detection rate of the parasite from stool samples by FECT, STST, BCT, APC and RT-PCR was 2.0% (17/844), 4.0% (34/844), 10.2% (86/844), 10.9% (92/844) and 28.8% (243/844), respectively. The highest detection rate (37.8%; 319/844) of S. stercoralis was recorded by a combination of BCT, APC, and RT-PCR followed by a combination of STST, BCT, APC and RT-PCR (37.3%; 315/844). The sensitivity of FECT, STST, BCT, APC and RT-PCR against the composite reference was 5.2%, 10.3%, 26.4%, 28.0% and 73.9%, respectively. The diagnostic agreements of RT-PCR, APC, BCT, STST and FECT with the composite reference in detection of S. stercoralis were substantial (0.775), fair (0.321), fair (0.305), slight (0.123), and slight (0.062), respectively. Conclusion: RT-PCR detected the highest number of S. stercoralis infections. A combination of RT-PCR with APC and/or BCT better detected S. stercoralis from stool samples compared to other combinations or single diagnostic methods. Therefore, RT-PCR and combination of RT-PCR with APC and/or BCT diagnostic methods should be advocated for detection of S. stercoralis infection.The fund obtained from Bahir Dar University and Mundo Sano Foundation was used for data collection and laboratory detection purpose.N

Prevalence of Strongyloides stercoralis infection and associated clinical symptoms among schoolchildren living in different altitudes of Amhara National Regional State, northwest Ethiopia

Background: Strongyloides stercoralis is a parasite that causes strongyloidiasis in humans. It is prevalent in the tropics and sub-tropics where poor sanitation is a common problem. The true prevalence of S. stercoralis in Ethiopia is underestimated due to the lack of a "Gold" standard diagnostic method. Moreover, its prevalence across altitudinal gradient in Amhara Region has not been studied. Methods: A cross-sectional study was conducted among 844 schoolchildren in Amhara Region from April to December 2019. A stool sample was collected from each study participant and processed using formol ether concentration technique (FECT), spontaneous tube sedimentation technique (STST), Baermann concentration technique (BCT), agar plate culture (APC) and real-time polymerase chain reaction (RT-PCR). Data were entered using EpiData and analyzed by SPSS version 23 statistical software. Prevalence of S. stercoralis infection was determined using a single diagnostic technique and combination of techniques. Association of clinical variables with S. stercoralis infection was assessed by logistic regression and independent variables with p<0.05 were considered statistically significant. Results: Prevalence of soil-transmitted helminths (STHs) and S. mansoni infections was 38.0% and 20.4%, respectively. Among STHs, the prevalence of hookworm infection was 32.8%. Prevalence of S. stercoralis infection was 39.0%, 28.8%, 10.9%, 10.3%, 4.0% and 2.0% by the respective, combinations of the five methods, RT-PCR, APC, BCT, STST and FECT. The highest prevalence rates, 48.2%, 45.0% and 41.1% of S. stercoralis were recorded in the age group of 12-14 years, males and rural dwellers, respectively. Prevalence rates of S. stercoralis infection in highland, semi-highland and lowland areas were 40.4%, 41.8% and 25.9%, respectively. Having abdominal pain (AOR = 2.48; 95% CI:1.65-3.72), cough (AOR = 1.63;95%CI:1.09-2.42), urticaria (AOR = 2.49;95%CI:1.50-4.01) and being malnourished (AOR = 1.44;95%:1.10-2.01) were significantly associated with strongyloidiasis. Conclusion: Prevalence of S. stercoralis infection was high and varied across different altitudes in Amhara Region. Some clinical syndromes were found to be significantly associated with S. stercoralis infection. Therefore, proper diagnosis and preventive strategies against S. stercoralis infection are highly recommended to be devised and implemented in Amhara Region.This study was funded by the Bahir Dar University (https://www.bdu.edu.et), Mundo Sano Foundation (https://www.mundosano.org/es/) and the National Center of Tropical Medicine, from the Institute of Health Carlos III (https://www.isciii.es/QuienesSomos/CentrosPropios/CNMT/Paginas/default.aspx), which supported the implementation by providing material support. We received materials and reagents but not a specific budget from funders to this project. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.S

Comparative whole-genome sequence analysis of Mycobacterium tuberculosis isolated from pulmonary tuberculosis and tuberculous lymphadenitis patients in Northwest Ethiopia

Background: Tuberculosis (TB), caused by the Mycobacterium tuberculosis complex (MTBC), is a chronic infectious disease with both pulmonary and extrapulmonary forms. This study set out to investigate and compare the genomic diversity and transmission dynamics of Mycobacterium tuberculosis (Mtb) isolates obtained from tuberculous lymphadenitis (TBLN) and pulmonary TB (PTB) cases in Northwest Ethiopia. Methods: A facility-based cross-sectional study was conducted using two groups of samples collected between February 2021 and June 2022 (Group 1) and between June 2020 and June 2022 (Group 2) in Northwest Ethiopia. Deoxyribonucleic acid (DNA) was extracted from 200 heat-inactivated Mtb isolates. Whole-genome sequencing (WGS) was performed from 161 isolates having ≥1 ng DNA/μl using Illumina NovaSeq 6000 technology. Results: From the total 161 isolates sequenced, 146 Mtb isolates were successfully genotyped into three lineages (L) and 18 sub-lineages. The Euro-American (EA, L4) lineage was the prevailing (n = 100; 68.5%) followed by Central Asian (CAS, L3, n = 43; 25.3%) and then L7 (n = 3; 2.05%). The L4.2.2.ETH sub-lineage accounted for 19.9%, while Haarlem estimated at 13.7%. The phylogenetic tree revealed distinct Mtb clusters between PTB and TBLN isolates even though there was no difference at lineages and sub-lineages levels. The clustering rate (CR) and recent transmission index (RTI) for PTB were 30 and 15%, respectively. Similarly, the CR and RTI for TBLN were 31.1 and 18 %, respectively. Conclusion and recommendations: PTB and TBLN isolates showed no Mtb lineages and sub-lineages difference. However, at the threshold of five allelic distances, Mtb isolates obtained from PTB and TBLN form distinct complexes in the phylogenetic tree, which indicates the presence of Mtb genomic variation among the two clinical forms. The high rate of clustering and RTI among TBLN implied that TBLN was likely the result of recent transmission and/or reactivation from short latency. Hence, the high incidence rate of TBLN in the Amhara region could be the result of Mtb genomic diversity and rapid clinical progression from primary infection and/or short latency. To validate this conclusion, a similar community-based study with a large sample size and better sampling technique is highly desirable. Additionally, analysis of genomic variants other than phylogenetic informative regions could give insightful information. Combined analysis of the host and the pathogen genome (GXG) together with environmental (GxGxE) factors could give comprehensive co-evolutionary information.The sample collection was funded by the Institute of Biotechnology, Bahir Dar University through the EN mega project. The Mtb culture and identification-related lab supply were supported by Amhara Public Health Institute, Bahir Dar Ethiopia. The whole-genome sequencing (WGS) and publication fee was covered by the National Center of Microbiology, Institute of Carlos III, Madrid, Spain. International Federation for Clinical Chemistry (IFCC) gave financial support to DM through the IFCC Professional Scientific Exchange Programme (PSEP) for 3-month WGS laboratory work.S

Cutaneous leishmaniasis in a newly established treatment centre in the Lay Gayint district, Northwest Ethiopia

Background: Cutaneous leishmaniasis (CL) is a neglected tropical disease that primarily affects the most vulnerable populations. In Ethiopia, where this study took place, CL is an important health problem, however, the incidence of CL is poorly monitored. Objectives: This study took place in a recently established CL treatment centre, at Nefas Mewcha Hospital, Lay Gayint. This area was considered to be endemic for CL, however, no cases of CL from Lay Gayint had previously been officially reported to the Amhara Regional Health Bureau. Methods: Following a CL awareness campaign, a retrospective data review was performed of patients presenting to this centre between July 2019 and March 2021. Basic demographic and clinical data were collected by a nurse and recorded in the logbook of the CL treatment centre. Results: Two hundred and one patients presented for diagnosis and treatment. The age of the patients ranged from 2 to 75 years and 63.2% were males. Most patients were between 10- and 19-years-old. The majority (79.1%) of the patients presented with localised cutaneous leishmaniasis and 20.9% with mucocutaneous leishmaniasis. 98% of the patients tested positive for Leishmania parasites by microscopy. Conclusions: This work underpinned how CL is a major public health problem in the Lay Gayint district. It also shows that raising awareness about CL in the community and providing diagnosis and treatment encouraged patients to travel to seek diagnosis and treatment

Mycobacterium tuberculosis Sub-Lineage 4.2.2/SIT149 as Dominant Drug-Resistant Clade in Northwest Ethiopia 2020-2022: In-silico Whole-Genome Sequence Analysis

Introduction: Drug resistance (DR) in Mycobacterium tuberculosis complex (MTBC) is mainly associated with certain lineages and varies across regions and countries. The Beijing genotype is the leading resistant lineage in Asia and western countries. M. tuberculosis (Mtb) (sub) lineages responsible for most drug resistance in Ethiopia are not well described. Hence, this study aimed to identify the leading drug resistance sub-lineages and characterize first-line anti-tuberculosis drug resistance-associated single nucleotide polymorphisms (SNPs). Methods: A facility-based cross-sectional study was conducted in 2020-2022 among new and presumptive multidrug resistant-TB (MDR-TB) cases in Northwest Ethiopia. Whole-genome sequencing (WGS) was performed on 161 isolates using Illumina NovaSeq 6000 technology. The SNP mutations associated with drug resistance were identified using MtbSeq and TB profiler Bioinformatics softwares. Results: Of the 146 Mtb isolates that were successfully genotyped, 20 (13.7%) harbored one or more resistance-associated SNPs. L4.2.2.ETH was the leading drug-resistant sub-lineage, accounting for 10/20 (50%) of the resistant Mtb. MDR-TB isolates showed extensive mutations against first-line anti-TB drugs. Ser450Leu/(tcg/tTg) for Rifampicin (RIF), Ser315Thr/(agc/aCc) for Isoniazid (INH), Met306Ile/(atg/atA(C)) for Ethambutol (EMB), and Gly69Asp for Streptomycin (STR) were the leading resistance associated mutations which accounted for 56.5%, 89.5%, 47%, and 29.4%, respectively. The presence of both clustered and non-clustered drug resistance (DR) isolates indicated that the epidemics is driven by both new DR development and acquired resistance. Conclusion: The high prevalence of drug-resistant TB due to geographically restricted sub-lineages (L4.2.2.ETH) indicates the ongoing local micro epidemics. The Mtb drug resistance surveillance system must be improved. Further evolutionary analysis of L4.2.2.ETH strain is highly desirable to understand evolutionary forces that leads L4.2.2.ETH in to high level DR and transmissible sub-lineage.Sample collection was funded by the Institute of Biotechnology, Bahir Dar University, through the Endalkachew Nibret Mega Project. The Mtb culture and identification-related laboratory supply was supported by the Amhara Public Health Institute, Bahir Dar, Ethiopia. Whole-genome sequencing was performed with the great support of National Center of Microbiology, Institute of Carlos III, Madrid, Spain. The International Federation for Clinical Chemistry (IFCC) provided financial support to Daniel Mekonnen through the IFCC Professional Exchange Program (PEP) for three months stay in Madrid Spain for conducting the WGS analysis.S

Seroprevalence of syphilis and its risk factors among pregnant women attending antenatal care at Felege Hiwot Referral Hospital, Bahir Dar, northwest Ethiopia: a cross-sectional study

Abstract Objective The objective of this study was to investigate the seroprevalence and associated risk factors of syphilis among pregnant women attending antenatal care at Felege Hiwot Referral Hospital, northwest Ethiopia. Result Of the total 384 screened women for syphilis, 10 (2.6%) were found to be seropositive for Treponema pallidum. The odds of infection were about ten times (COR = 9.77, p = 0.002) higher in divorced women than in married women. The likelihood of syphilis was almost three times higher among rural residents compared to urban residents (COR = 3.48, p = 0.079). The likelihood of being infected with syphilis was about five times higher (COR = 5.25, p = 0.018) in women who had prior history of multiple sexual partners. The risk of syphilis was 4.42 (p = 0.071) and 2.67 fold (p = 0.226) greater in women with previous history of abortion and other sexually transmitted diseases (STDs), respectively. It can be concluded that a relatively low seroprevalence of syphilis was observed among the study subjects. Nevertheless, health promotion activity directed at raising the awareness of the community towards the modes of transmission of syphilis and its health impact is important in the prevention of the disease in pregnant women

Prevalence of intestinal helminthic infections and malnutrition among schoolchildren of the Zegie Peninsula, northwestern Ethiopia

Summary: A cross-sectional study involving 408 schoolchildren was performed from November 2013 to February 2014 to assess the prevalence of intestinal helminthic infections and malnutrition among schoolchildren of the Zegie Peninsula, northwestern Ethiopia. A structured questionnaire was used to gather data on the demographic and risk factors associated with intestinal helminthic infections. Stool samples were collected and examined for helminth eggs using the formalin–ether concentration technique. In addition, anthropometric measurements were performed to assess the nutritional status of the schoolchildren. The overall prevalence of intestinal helminthic infections in the study area was 69.1%. Single, double, triple, and quadruple infections were 42.2%, 18.4%, 5.9%, and 0.7%, respectively. The prevalence of hookworm, Schistosoma mansoni, Ascaris lumbricoides, Trichuris trichiura, Hymenolepis nana, and Strongyloides stercoralis infections were 43.4%, 29.9%, 12.7%, 10%, 4.6%, and 0.7%, respectively. The prevalence of malnutrition in terms of stunting, being underweight, and wasting were 15.3%, 18%, and 27.6%, respectively. However, no significant association was found between intestinal helminthic infections and stunting, being underweight and wasting (p > 0.05). It can be concluded that intestinal helminthic infections, specifically infections due to hookworm and S. mansoni, are important health problems among schoolchildren of the Zegie Peninsula. Keywords: Ethiopia, Intestinal helminthes, Malnutrition, Schoolchildren, Zegie Peninsul

Prevalence of Schistosoma mansoni and S. haematobium in Snail Intermediate Hosts in Africa: A Systematic Review and Meta-analysis

Background. Schistosomiasis is caused by Schistosoma mansoni and S. haematobium in Africa. These schistosome parasites use freshwater snail intermediate hosts to complete their lifecycle. Varied prevalence rates of these parasites in the snail intermediate hosts were reported from several African countries, but there were no summarized data for policymakers. Therefore, this study was aimed to systematically summarize the prevalence and geographical distribution of S. mansoni and S. haematobium among freshwater snails in Africa. Methods. Literature search was carried out from PubMed, Science Direct, and Scopus which reported the prevalence of S. mansoni and S. haematobium among freshwater snails in Africa. The pooled prevalence was determined using a random-effect model, while heterogeneities between studies were evaluated by I2 test. The meta-analyses were conducted using Stata software, metan command. Results. A total of 273,643 snails were examined for the presence of S. mansoni and S. haematobium cercaria in the eligible studies. The pooled prevalence of schistosome cercaria among freshwater snails was 5.5% (95% CI: 4.9–6.1%). The pooled prevalence of S. mansoni and S. haematobium cercaria was 5.6% (95% CI: 4.9–6.3%) and 5.2% (95% CI: 4.6–5.7%), respectively. The highest pooled prevalence was observed from Nigeria (19.0%; 95% CI: 12.7–25.3%), while the lowest prevalence was reported from Chad (0.05%; 95% CI: 0.03–0.13). Higher prevalence of schistosome cercaria was observed from Bulinus globosus (12.3%; 95% CI: 6.2–18.3%) followed by Biomphalaria sudanica (6.7%; 95% CI: 4.5–9.0%) and Biomphalaria pfeifferi (5.1%; 95% CI: 4.1–6.2%). The pooled prevalence of schistosome cercaria obtained using PCR was 26.7% in contrast to 4.5% obtained by shedding cercariae. Conclusion. This study revealed that nearly 6% of freshwater snails in Africa were infected by either S. haematobium or S. mansoni. The high prevalence of schistosomes among freshwater snails highlights the importance of appropriate snail control strategies in Africa