Plasmodium falciparum histidine-rich protein 2 and 3 genes deletion in global settings (2010-2021): a systematic review and meta-analysis.

BACKGROUND: The usefulness of histidine-rich protein-2/3 (HRP2/3)-based rapid diagnostic tests of malaria due to Plasmodium falciparum has been threatened by the appearance of mutant PfHRP2/3 genes. This study was undertaken to determine the global pooled estimates of PfHRP2/3gene deletions. METHODS: Relevant publications were identified from electronic databases such as; PubMed, EMBASE, and MEDLINE online. Besides, all the relevant literatures were retrieved through Google and Google Scholar. STATA software was used for data analysis. The pooled estimates were calculated using random effect model. The summary estimates were presented using forest plots and tables. RESULTS: A total of 27 studies were included in the systematic review. However, only 24 and 17 studies were included for PfHRP2 and 3 gene deletion meta-analysis, respectively. The prevalence of PfHRP2 gene deletion across the individual studies ranged from the highest 100% to the lowest 0%. However, the meta-analysis result showed that the global pooled prevalence of PfHRP2 and PfHRP3 gene deletions were 21.30% and 34.50%, respectively. The pooled proportion of PfHRP2 gene deletion among false negative PfHRP2-based RDTs results was found to be 41.10%. The gene deletion status was higher in South America and followed by Africa. The pooled estimate of PfHRP2 gene deletion among studies, which did not follow the WHO PfHRP2/3 gene deletion analysis protocol was higher than their counter parts (21.3% vs 10.5%). CONCLUSIONS: This review showed that there is a high pooled prevalence of PfHRP2/3 gene deletions in Plasmodium falciparum confirmed isolates and also a high proportion of their deletions among false-negative malaria cases using PfHRP2-based RDT results. Hence, malaria diagnosis based on PfHRP2-based rapid tests seems to be less sensitive and warrants further evaluation of PfHRP2/3 gene deletions.

Efficacy of a single dose versus a multiple dose regimen of Mebendazole against hookworm infections among school children: a randomized open-label trial.

BACKGROUND: Despite the existence of a population-based control program using single dose albendazole or mebendazole as a preventive chemotherapy, hookworm transmission remains high. It causes a negative impact on the growth and school performance of children. In connection to this preventive chemotherapy, different studies produced conflicting results. This study aimed at evaluating the efficacy of single (500 mg) versus multiple doses (100 mg twice a day during three consecutive days) of mebendazole against hookworm infections among school-aged children. METHODS: This randomized open-label clinical trial took place among school-aged children (6-14 years old) in Burie and Debre Elias towns, Northwest Ethiopia. Using simple randomization, eligible hookworm-positive children were allocated (1:1) to either a single or multiple dose treatment arms. Stool samples were collected and processed using McMaster method at baseline and follow-up period (14-21 days after treatment). Only laboratory technicians were blinded. The cure and egg reduction rates were the primary and secondary therapeutic outcome measures against hookworm infections, respectively. An independent t-test was used to compare group means, and logistic regression was used to calculate odds ratio (OR). P-value < 0.05 at 95% CI was considered statistically significant. RESULT: One hundred eight children, 54 in each treatment arm had completed baseline data and received allocated treatment. One hundred three children had completed follow-up data records and included for the final efficacy analysis. Cure rate against hookworm was significantly higher in the multiple dose (96.1%) than in the single dose (30.8%) with OR = 55.125; 95% CI: 11.92-254.9; P < 0.001. The egg reduction rate in the multiple dose treatment arm (99.5%) was also significantly higher than in the single dose arm (68.9%) with difference t (101) =5.38; 95% CI 230.95-505.36; P < 0.001. CONCLUSION: The single dose regimen of mebendazole for the treatment of hookworm infections showed poor cure and egg reduction rates, while the multiple doses revealed satisfactory. Although multiple dose regimen administration is a bit more complex than the single dose, we strongly encourage replacing it with multiple dose regimen during deworming programs in hookworm endemic areas. TRIAL REGISTRATION: This trial is retrospectively registered in www.pactr.org, number PACTR201911466695052 on November 26, 2019.

Barriers of persistent long-lasting insecticidal nets utilization in villages around Lake Tana, Northwest Ethiopia: a qualitative study.

BACKGROUND: Malaria remains a major public health problem in Ethiopia. The use of LLINs is an effective approach to reducing transmission. Persistent use of LLINs is determined by numerous factors. Quantitative studies have assessed LLIN ownership and utilization, but the behavioral, socio-cultural, socioeconomic and net distribution contexts that impact their use have not been examined in depth. This study aimed to explore barriers of persistent LLIN use among communities around Lake Tana. METHODS: Twenty-three community residents who owned LLINs (15) or not (8) during the study period and 38 key informants were interviewed from April to June 2017. Phenomenological study was employed to explore the local contexts and factors that influence persistent use of LLINs. Individuals were purposefully selected to capture different views. Community residents were selected based on their permanent residence and LLIN use experience. Key informants were health extension workers, local leaders, students, and health professionals. The data were managed using QSR International NVivo Version 10 software and coded, and themes were identified. RESULTS: Killing ability of nets against arthropods other than mosquitoes reportedly made use of LLINs a favored malaria prevention method despite their ineffectiveness after 3 months. Conical nets were preferred due to their compatibility with varied sleeping structures. Numerous factors influenced persistent use, notably erroneous perceptions about LLINs, malaria and mosquitoes; bedbug infestation; inconvenience; unintended uses; distribution problem of nets; and socio-cultural and economic factors. Unintended uses were often associated with local needs and seldom linked with social issues and deficiencies in information about malaria and LLINs. Collateral benefits were considered important, principally in terms of disinfestation of bedbugs. CONCLUSIONS: Non-persistent LLIN use was associated with inconvenient bed net design and early damage; non-potency of the insecticide against other arthropods; facilitation of bedbug infestation; unintended uses; wrong perceptions about malaria, mosquitoes, and LLINs; and inadequate follow-up regarding LLINs utilization. Distribution of conical nets and provision of adequate information on LLINs and malaria may promote persistent use. Using an insecticide that also kills arthropods other than mosquitoes may reduce unintended uses and increase persistent use.

Travel to farms in the lowlands and inadequate malaria information significantly predict malaria in villages around Lake Tana, northwest Ethiopia: a matched case-control study.

BACKGROUND: In Ethiopia, malaria has declined in the last decade; only a small number of cases have been reported, primarily from hotspots. The contribution of house proximity to water bodies and the role of migration in malaria transmission has not yet been examined in detail in northwest Ethiopia. Individual and household-level environmental and socio-demographic drivers of malaria heterogeneity were explored contextually in meso-endemic villages around Lake Tana, northwest Ethiopia. METHODS: A health facility-based paired age-sex matched case-control study involving 303 matched pairs was undertaken from 10 October 2016, to 30 June 2017. Geo-referencing of case households, control households, proximate water bodies, and health centres was carried out. A pretested and structured questionnaire was used to collect data on socio-demography, household assets, housing, travel history, and malaria intervention measures. Medians (interquartile range) were computed for continuous variables. Pearson's Chi square/Fisher's exact test was used to detect significant differences in proportions. Principal component analysis was performed to estimate household wealth. Stratified analysis was used to confirm confounding and interaction. A multivariable conditional logistic regression model was used to detect risk factors for malaria. RESULTS: Of 303 malaria cases, 59 (19.5% [15.4-24.3]) were imported malaria cases whereas 244 (80.5% [75.7-84.6]) were locally acquired malaria cases. In bivariate analysis, marital status, educational status, and bed net ownership were significantly associated with malaria cases. In multivariable adjustment, travel to malarious lowlands in the preceding month (adjusted mOR = 7.32; 95% CI 2.40-22.34), household member's travel to malarious lowlands (adjusted mOR = 2.75; 95% CI 1.02-7.44), and inadequate health information on malaria (adjusted mOR = 1.57; 95% CI 1.03-2.41) were predictors of malaria. Stratified analysis confirmed that elevation of households and travel to malarious lowlands were not effect modifiers. Travel to malarious lowlands had a confounding effect on malaria but elevation of households did not. CONCLUSIONS: In this study, travel to farms in the lowlands and inadequate health information on malaria were risk factors for malaria in villages around Lake Tana. This evidence is critical for the design of improved strategic interventions that consider imported malaria cases and approaches for accessing health information on malaria control in northwest Ethiopia.

Genetic diversity, multiplicity of infection and population structure of Schistosoma mansoni isolates from human hosts in Ethiopia.

BACKGROUND: Human intestinal schistosomiasis caused by Schistosoma mansoni and urinary schistosomiasis caused by Schistosoma haematobium are endemic in Ethiopia. Although schistosomes look morphologically uniform, there is variation in infectivity, egg productivity and virulence due to variation in their genetic make. Knowing the genetic diversity and population structure of S. mansoni isolates will enable to understand and consider the possible variability in terms of infectivity, egg productivity and virulence. METHODS: Between 2010 and 2011, genetic diversity and population structure of Schistosoma mansoni isolates from four endemic areas of Ethiopia was assessed using previously published 11 polymorphic microsatellite loci. Miracidia were hatched from eggs of S. mansoni collected from stools of human subjects residing in Kemissie, Wondo Genet, Ziway and Sille-Elgo villages. DNA was extracted from single miracidium and PCR was run following standard protocol. Allelic polymorphism and population genetic structure was analyzed using different software. RESULT: At a population level (i.e. different villages), the mean number of alleles per locus, allelic richness, expected heterozygosity in Hardy-Weinberg equilibrium and pairwise F ST values ranged from 8.5 to 11.5, 3.46-20.8, 0.66-0.73 and 3.57-13.63%, respectively. All analyzes on population genetic structure reveals strong genetic structuration corresponding to the four sampled villages. At infrapopulation level (i.e. different hosts) the mean number of alleles per locus, allelic richness, expected heterozygosity in Hardy-Weinberg equilibrium and F IS values ranged from 3.09 to 7.55, 1-1.96, 0.59-0.73 and 0.1763-0.4989, respectively. Mean estimated genetically unique adult worm pairs within hosts ranged from 66 to 92% revealing the occurrence of infection of a single host with genetically unique multiple S. mansoni strains. The data also indicated the occurrence of genetic variation within inter- and intra-hosts. CONCLUSION: High level of genetic diversity and significant population differentiation characterized the S. mansoni isolates of Ethiopia. These results are quite different from previous studies demonstrating that it is difficult to generalize schistosome transmission patterns because epidemiological situation tends to vary. These are important factors to be considered in relation with morbidity, drug resistance or vaccine development.

Detection of mixed infection level of Plasmodium falciparum and Plasmodium vivax by SYBR Green I-based real-time PCR in North Gondar, north-west Ethiopia.

BACKGROUND: Malaria is caused by five Plasmodium species and transmitted by anopheline mosquitoes. It occurs in single and mixed infections. Mixed infection easily leads to misdiagnosis. Accurate detection of malaria species is vital. Therefore, the study was conducted to determine the level of mixed infection and misdiagnosis of malaria species in the study area using SYBR Green I-based real time PCR. METHODS: The study was conducted in seven health centres from North Gondar, north-west Ethiopia. The data of all febrile patients, who attended the outpatient department for malaria diagnosis, from October to December 2013, was recorded. Dried blood spots were prepared from 168 positive samples for molecular re-evaluation. Parasite DNA was extracted using a commercial kit and Plasmodium species were re-evaluated with SYBR Green I-based real time PCR to detect mixed infections and misdiagnosed mono-infections. RESULTS: Among 7343 patients who were diagnosed for malaria in six study sites within the second quarter of the Ethiopian fiscal year (2013) 1802 (24.54%) were positive for malaria parasite. Out of this, 1,216 (67.48%) Plasmodium falciparum, 553 (30.68%) Plasmodium vivax and 33 (1.8%) mixed infections of both species were recorded. The result showed high prevalence of P. falciparum and P. vivax, but very low prevalence of mixed infections. Among 168 samples collected on dried blood spot 7 (4.17%) were P. vivax, 158 (94.05%) were P. falciparum and 3 (1.80%) were mixed infections of both species. After re-evaluation 10 (5.95%) P. vivax, 112 (66.67%) P. falciparum, 21 (12.50%) P. falciparum + P. vivax mixed infection, and 17 (10.12%) Plasmodium ovale positive rate was recorded. The re-evaluation showed high level of mixed infection, and misdiagnosis of P. ovale and P. vivax. CONCLUSIONS: The result shows that P. falciparum prevalence is higher than P. vivax in the study area. The results, obtained from SYBR Green I-based real time PCR, indicated that the diagnosis efficiency of microscopy is very low for species-specific and mixed infection detection. Therefore, real time PCR-based species diagnosis should be applied for clinical diagnosis and quality control purposes in order to prevent the advent of drug resistant strains due to misdiagnosis and mistreatment.

Assessment of Urogenital Schistosomiasis Knowledge, Attitudes and Practices Among Abobo Communities in Gambella Regional State, Southwestern Ethiopia.

Background: Knowledge about community awareness and practices related to a disease, among other things, helps to plan relevant control strategies. This study assessed the knowledge, attitude, and practices (KAPs) of the community in Abobo district of Gambella Regional State in Southwestern Ethiopia about urogenital schistosomiasis (UGS), which is endemic to the region. Methods: A community-based cross-sectional study was conducted in 2022. A pretested structured questionnaire was administered. Multivariable logistic regression was used to examine factors associated with KAPs variables. Results: Most study participants (90.6%) responded they have previously heard of UGS. Over 95% of the participants knew at least 1 symptom of UGS and 30% knew the transmission cycle. About 15.9% and 26.8% knew keeping environmental hygiene and refraining from using cercariae infested water, as preventive methods, respectively. Over half of the participants (50.1%) disagreed or did not know that urinating close to the river/dam water contributes to transmission, and almost all participants had contact with the dam/river water in one way or another. Education status and history of having the infection were the key significant correlates of most KAPs variables. Conclusion: Most of the community members had poor knowledge and attitudes about the mode of transmission and preventive measures of UGS. Thus, besides school-based mass drug administration (MDA), UGS control efforts in the region should incorporate health education in conjunction with safe water supplies, and provision of sanitary facilities to effectively reduce the transmission of the disease.

In vitro antileishmanial activities of hydro-methanolic crude extracts and solvent fractions of Clematis simensis fresen leaf, and Euphorbia abyssinica latex.

As a result of increasing drug resistance, crossover resistance development, prolonged therapy, and the absence of different agents with innovative methods for implementation, the efficacy of recent antileishmanial medications is severely declining. So, it is vital to look for other medications from botanical remedies that have antileishmanial activity. The latex of Euphorbia abyssinica (E abyssinica) and the leaves of Clematis simensis fresen (C simensis) were macerated in methanol (80%). In vitro antileishmanial activity of the preparation was tried on promastigotes of Leishmania aethiopica (L aethiopica) and Leishmania donovani (L donovani) using resazurin assay, and fluorescence intensity was measured. One percent of dimethyl sulfoxide (DMSO) and media as negative control and amphotericin B as positive control were used. Additionally, hemolytic & phytochemical tests of the preparation were done. The mean and standard errors of each extract were evaluated and interpreted for statistical significance using one-way analysis of variance. From sigmoidal dose-response curves of % inhibition, half maximal inhibitory concentration (IC50) values were determined by GraphPad Prism and Microsoft Excel; outcomes were presented as mean ±â€…standard error of mean of triplicate trials. P < .05 was statistical significance. The phytochemical screening of C simensis and E abyssinica confirmed the existence of steroids, phenols, tannins, saponins, alkaloids, terpenoids, flavonoids and glycosides. C simensis possesses antileishmanial activity with IC50 outcomes of 46.12 ±â€…0.03 and 8.18 ±â€…0.10 µg/mL on the promastigotes of L aethiopica and L donovani, respectively. However, E abyssinica showed stronger activity with IC50 outcomes of 16.07 ±â€…0.05 µg/mL and 4.82 ±â€…0.07 µg/mL on L aethiopica and L donovani, respectively. C simensis and E abyssinica have a less hemolytic effect on human red blood cells at low concentrations. The outcomes from this investigation demonstrated that the preparation of C simensis and E abyssinica indicated significant antileishmanial activity. Therefore, further in vivo assessment of antileishmanial, cytotoxicity activity and quantitative identification of secondary metabolites are highly recommended.

Ecology and Infection Status of Sand Flies in Rural and Urban Cutaneous Leishmaniasis Endemic Areas in Northwest Ethiopia.

Cutaneous leishmaniasis (CL) caused by Leishmania aethiopica is transmitted by Phlebotomus longipes in northern Ethiopia. No studies have been conducted to investigate the transmission dynamics of CL, despite its high endemicity in both rural and urban settings. Evidence on the ecology and behavior of the vector from this area are required to develop integrated disease control strategies. Sand flies were collected in the dry and wet seasons in 2021 in CL-endemic rural Gindmeteaye and urban Addis-Alem in northwest Ethiopia. Trapping was performed with sticky and Centers for Disease Control and Prevention (CDC) light traps in three habitats, including inside patients' houses, peridomestic areasand in caves/rocky areas. Sand flies were morphologically identified to species level. Female Phlebotomus species were categorized according to blood feeding status and tested by spliced-leader (SL-) ribonucleic acid (RNA) polymerase chain reaction (PCR) to screen for Leishmania infection. Of 1161 sand flies, the majority (77%) were P. longipes, six (0.5%) were P. orientalis and the remaining were Sergentomyia. The abundance of the 430 female P. longipes was significantly linked to seasonality (p < 0.001), with the majority in the dry season occurring in the outdoor rocky (37%) and peridomestic (34%) sites, while, in the wet season, most (62%) were captured indoors. This seasonality was more pronounced in rural Gindmeteaye, where housing construction is poor. The number of blood-fed and gravid P. longipes was significantly higher in the wet (31%; 22%), compared to the dry season (13%; 8%), and their proportion was highest indoors. Eighteen (4%) female P. longipes were Leishmania positive, with highest infection prevalence in caves (7% compared to 3% indoors, p = 0.022), and in the dry season (6%, p < 0.001). Phlebotomus orientalis specimens were all captured in May in rural Gindmeteaye, five indoors and one in a peridomestic site. Further research should be conducted to investigate the absolute contribution of humans and indoor transmission to the transmission cycle of CL. Inhabitants of endemic villages should be made aware that evening outdoor activities near caves may increase their exposure to infectious sand flies. Whether P. orientalis can breed and become infected at high altitudes should be further studied.

Current Status of Urinary Schistosomiasis Among Communities in Kurmuk District, Western Ethiopia: Prevalence and Intensity of Infection.

Background: Schistosomiasis is a highly prevalent but neglected tropical disease, particularly in sub-Saharan Africa. In Ethiopia, urogenital schistosomiasis due to Schistosoma haematobium has been known to be endemic in several lowland areas. This study was designed to determine the current prevalence and intensity of the urogenital schistosomiasis among communities in Kurmuk District, western Ethiopia. Methods: Urine filtration technique and urine dipstick test were used to screen for S. haematobium eggs and hematuria, respectively. The data were analyzed with SPSS version 23. Logistic regression and odds ratio were used to measure associations and strength between prevalence, intensity, and independent variables. P-values <.05 at 95% CI were considered statistically significant. Results: The overall prevalence of S. haematobium infection as determined by urine filtration was 34.2% (138/403). In bivariate analysis, the most infected (45.4%) age groups were 5 to 12 years (odds ratio [OR] = 4.16, 95% CI: 1.36-12.67), followed by 13 to 20 years (OR = 3.23, 95% CI: 1.01-10.35) with higher significant mean egg count (MEC). The mean egg intensity ranged from 2.39 in Ogendu (CI: 1.05-3.72) to 14.1 in Dulshatalo (CI: 4.98-23.12) villages. The main predictor of infection was swimming habits (adjusted odds ratio [AOR] = 2.43 [CI: 1.19-4.94]). The prevalence of hematuria was 39.2% (158/403), the odds being 2.64 times higher among participants who resided in Dulshatalo than those who resided in Kurmuk (AOR 2.64 [95% CI: 1.43-4.87], P = .004). Conclusion: To reduce the infection and interrupt transmission, the PC in place in the area using PZQ should be strengthened and continued, alongside with provision of sanitary facilities, safe alternative water supplies and health education. The Federal Ministry of Health of Ethiopia should also collaborate with the health authorities of the Sudan government for the control of trans-boundary transmission of the disease as the transmission foci are shared between the 2 countries.

Liver Enzymes and Lipid Profile of Malaria Patients Before and After Antimalarial Drug Treatment at Dembia Primary Hospital and Teda Health Center, Northwest, Ethiopia.

Background: Infection with malaria in humans involves liver cell destruction, which alters the levels of liver enzymes and lipid profiles. A number of studies have been conducted to address the impact of malaria on liver enzymes and lipid profiles but no studies were addressed after antimalarial treatment in Ethiopia. This study is intended to fill this gap. Methods: An observational cohort study was conducted at Dembia Primary Hospital and Teda Health Center, from June to August 2020. Eighty eight malaria infected study participants were recruited using random sampling techniques. Socio-demographic data, capillary and venous blood samples were collected. Assessment of liver enzymes and lipid profiles was done using Beckman Coulter DC-700 clinical chemistry analyzer. Data were entered using Epi-data and exported to SPSS version 20 for analysis. One way ANOVA, independent t-test, and paired t-test were used to compare the mean liver enzymes and lipid profile. p-value<0.05 was considered statistically significant. Results: Before anti-malaria treatment, among 88 study participants, elevated AST (87.5%), ALT (12.5%), ALP (43.2%), and TG (17.2%) and lower HDL (87.5%) and normal LDL and TC were observed. After treatment, 100% AST, ALT, HDL, and LDL and 92% ALP, 94.3% TC, and 86.4% TG levels were in the normal range. The mean level of AST and ALT increased while HDL decreased from low to higher density parasitaemia. Mean level of AST was significantly lower while ALT did not alter. HDL, LDL, and TC level were increased but statistically were insignificant (P>0.05). Conclusion: Malaria could be responsible for increased liver enzymes and certain lipids while decreasing some lipid profiles. After anti-malaria treatment, these parameters were reversed to normal from 86.4% to 100%. Hence, prompt treatment is important to improve liver enzymes and lipid profile impairment during malaria infection.

Phase 3 evaluation of an innovative simple molecular test for the diagnosis of malaria in different endemic and health settings in sub-Saharan Africa (DIAGMAL).

BACKGROUND: Rapid Diagnostic Tests (RDTs) have become the cornerstone for the management of malaria in many endemic settings, but their use is constrained for several reasons: (i) persistent malaria antigen (histidine-rich protein 2; HRP2) leading to false positive test results; (ii) hrp2 deletions leading to false negative PfHRP2 results; and (iii) limited sensitivity with a detection threshold of around 100 parasites/µl blood (pLDH- and HRP2-based) leading to false negative tests. Microscopy is still the gold standard for malaria diagnosis, and allows for species determination and quantitation, but requires trained microscopists, maintained microscopes and has detection limit issues. Consequently, there is a pressing need to develop and evaluate more sensitive and accurate diagnostic tests. To address this need we have developed a direct on blood mini PCR-NALFIA test that combines the benefits of molecular biology with low infrastructural requirements and extensive training. METHODS: This is a Phase 3 diagnostic evaluation in 5 African countries. Study sites (Sudan, Ethiopia, Burkina, Kenya and Namibia) were selected to ensure wide geographical coverage of Africa and to address various malaria epidemiological contexts ranging from high transmission to near elimination settings with different clinical scenarios and diagnostic challenges. Study participants will be enrolled at the study health facilities after obtaining written informed consent. Diagnostic accuracy will be assessed following the WHO/TDR guidelines for the evaluation of diagnostics and reported according to STARD principles. Due to the lack of a 100% specific and sensitive standard diagnostic test for malaria, the sensitivity and specificity of the new test will be compared to the available diagnostic practices in place at the selected sites and to quantitative PCR as the reference test. DISCUSSION: This phase 3 study is designed to validate the clinical performance and feasibility of implementing a new diagnostic tool for the detection of malaria in real clinical settings. If successful, the proposed technology will improve the diagnosis of malaria. Enrolment started in November 2022 (Kenya) with assessment of long term outcome to be completed by 2023 at all recruitment sites. TRIAL REGISTRATION: Pan African Clinical Trial Registry (www.pactr.org) PACTR202202766889963 on 01/02/2022 and ISCRTN (www.isrctn.com/) ISRCTN13334317 on 22/02/2022.

Evaluation of Hookworm Diagnosis Techniques from Patients in Debre Elias and Sanja Districts of the Amhara Region, Ethiopia.

BACKGROUND: Inappropriate diagnosis could intimidate the prevention and control of hookworm infection. Thus, this study was aimed at evaluating the performance of hookworm diagnosis methods. METHODS: An institution-based cross-sectional study was conducted from patients in Debre Elias and Sanja districts of the Amhara region, Ethiopia, from November 2019 to January 2020. The study subjects were selected conveniently. After the data was entered into Statistical Package for the Social Sciences (SPSS), sensitivity, specificity, predictive values, test accuracy, and agreement of the different hookworm diagnosis methods, namely, test tube flotation technique (TFT), McMaster (MM), formol-ether concentration (FEC), Kato-Katz (KK), and direct wet mount microscopy (DWMM), were calculated by using SPSS software. The composite reference standard (CRS) was used as the gold standard method. The kappa (κ) test was used to measure the level of agreement between diagnosis tests. RESULT: A total of 389 stool samples were collected from patients in the study. The overall prevalence of hookworm was 63.24%. The test tube flotation technique (TFT) was found to be the highest both in terms of sensitivity and diagnostic accuracy (100%). McMaster (MM) was the second most sensitive test (68.7%), followed by FEC (44.3%) and KK (38.2%). On the other hand, DWMM had the lowest sensitivity (37.4%), and its diagnostic accuracy was also the least (60%). Only TFT had a perfect agreement (agreement = 100%, kappa = 1) with the CRS. The sensitivity of DWMM, KK, and FEC showed a kind of linear function with the intensity of infection, but TFT and MM methods were not affected by the infection intensity. CONCLUSION: Hookworm is still a public health problem in the study area. TFT is by far more sensitive than MM, FEC, KK, and DWMM techniques.

In vivo and in vitro efficacy of a single dose of albendazole against hookworm infection in northwest Ethiopia: open-label trial.

BACKGROUND: Control of hookworm and other soil-transmitted helminth infections primarily relies on preventive chemotherapy using a single dose of albendazole/mebendazole drugs on high-risk groups. Herein, the efficacy of a single dose (400 mg) of albendazole (ALB) was investigated both in vivo and in vitro model in northwest Ethiopia. METHODS: An open-label, single-arm clinical trial was conducted to assess anti-hookworm effect of albendazole. Stool samples were collected and examined using McMaster and Harada-Mori filter paper culture. Eligible hookworm-infected patients were treated with a single dose of ALB. After 14-21 days post-treatment, stool samples were also taken again and re-examined using the abovementioned technique. Egg reduction rate (ERR) and larval motility were used as a therapeutic outcome measure. An independent t test was used to compare the mean difference in egg counts, and probit analysis was performed for calculating the lethal concentration dose of albendazole. P value < 0.05 at 95% CI was considered statistically significant. RESULTS: A total of 70 participants had completed the drug efficacy study. The efficacy of ALB against hookworm in terms of CR and ERR was 87% and 93%, respectively. Participants who had not eaten one or more hours prior to treatment had higher CR than those who had eaten within 1 h before treatment (97.4% vs 74.2%), while individuals with heavy infection intensity had a lower post-treatment ova clearing rate than those who were with light infection intensity (43% vs 94.6%). The in vitro larvicidal effect of ALB was 63-93% after applying 50-250 µg/ml concentration of ALB solution. The LC50 and LC99 were 152 µg/ml and 573 µg/ml, respectively. CONCLUSION: A single dose of albendazole was found to be effective for treating hookworm infections according to WHO anthelminthic evaluation standard in the study area. Preventive chemotherapy might therefore be extended to risk groups, with proper continuous monitoring of its efficacy to strengthen and keep the ongoing control and prevention measures one step ahead. TRIAL REGISTRATION: This trial is retrospectively registered with www.pactr.org , number PACTR202010511829332 on October 26, 2020.

Prevalence and factors associated with intestinal schistosomiasis and human fascioliasis among school children in Amhara Regional State, Ethiopia.

BACKGROUND: Schistosomiasis and fascioliasis are digenean parasitic infections and are among the neglected tropical diseases that have both medical and veterinary importance. They are found mainly in areas having limited access to safe water supply and improved sanitation. METHODS: A cross-sectional study was conducted to determine the prevalence of Schistosoma mansoni and Fasciola species infections and to identify associated risk factors among school children in Amhara Regional State, Ethiopia. Stool specimens were collected from 798 children (419 males, 379 females) and processed using Kato-Katz and formol-ether concentration techniques. A semi-structured questionnaire was used to collect socio-demographic and other exposure information to explore potential risk factors for the infections. RESULTS: The overall prevalence of S. mansoni and Fasciola species infections was 25.6% (95% confidence interval (CI): 22.5-28.6) and 5.5% (95% CI: 3.9-7.1), respectively. S. mansoni was present in all surveyed schools with the prevalence ranging from 12.8% (16/125; 95% CI = 5.6-20.0) to 39.7% (64/161; 95% CI = 32.2-47.2) while Fasciola species was identified in five schools with the prevalence ranging from 2.5% (4/160; 95% CI = 0.001-4.9) to 9.8% (13/133; 95% CI = 4.7-14.8). The prevalence of S. mansoni infection was significantly associated with swimming in rivers (Adjusted odds ratio (AOR): 1.79, 95% CI, 1.22-2.62; P=0.003), bathing in open freshwater bodies (AOR, 2.02; 95% CI, 1.39-2.94; P<0.001) and engaging in irrigation activities (AOR, 1.69; 95% CI, 1.19-2.39; P=0.004), and was higher in children attending Addis Mender (AOR, 2.56; 95% CI, 1.20-5.46; P=0.015 ) and Harbu schools (AOR, 3.53; 95% CI, 1.64-7.59; P=0.001). Fasciola species infection was significantly associated with consumption of raw vegetables (AOR, 2.47; 95% CI, 1.23-4.97; P=0.011) and drinking water from unimproved sources (AOR, 2.28; 95% CI, 1.11-4.70; P=0.026). CONCLUSION: Both intestinal schistosomiasis and human fascioliasis are prevalent in the study area, affecting school children. Behaviors and access to unimproved water and sanitation are among significant risk factors. The findings are instrumental for targeted interventions.

Hematological and Biochemical Profile of Patients Infected with Schistosoma mansoni in Comparison with Apparently Healthy Individuals at Sanja Town, Northwest Ethiopia: A Cross-Sectional Study.

BACKGROUND: Schistosomiasis is a parasitic disease that resides in the vascular system of vertebrates, causing a chronic, debilitating disease that affects more than 200 million people and 800,000 deaths per year in over 70 countries. This parasite causes liver dysfunction and disorders normal hematological and biochemical profiles in addition to portal vein hypertension syndrome, ascites, and liver fibrosis. The general objective of the current study is to assess hematological and biochemical profiles of patients infected with Schistosoma mansoni in comparison with apparently healthy individuals (control group) in Sanja town, northwest Ethiopia. METHOD: A comparative cross-sectional study was conducted from February to April 2019 among microscopically confirmed S. mansoni-infected patients attending Sanja hospital and apparently healthy (control group) from Sanja town community. A total of 220 participants, 110 from the S. mansoni-infected and 110 from the control group, were enrolled using convenient sampling technique. Three grams of stool and six milliliters of blood samples were collected from each study participant. Stool samples were processed using the Kato-Katz technique to determine infection and count parasite density. The blood sample was processed for the analysis of hematological and biochemical profiles using Cell Dyn 1800 (Abbot Hematology, IL, USA) and iChem535 chemistry analyzer, respectively. All data were analyzed using SPSS version 20, and P value less than 0.05 was taken as statistically significant. RESULTS: This study showed that the mean values of serum alanine aminotransferase, aspartate aminotransferase, total protein, total cholesterol, hemoglobin, mean corpuscular hemoglobin concentration, and total white blood cell count were different in the Schistosoma mansoni-positive group as compared with the control group with statistically significant value (P ≤ 0.05). However, the mean values of blood glucose, red blood cell, packed cell volume, and granulocyte count difference were not statistically significant (P ≥ 0.05). The mean value of hemoglobin, red blood cells, blood glucose, mean corpuscular hemoglobin concentration, total protein, total cholesterol, and total white blood cell was significantly dropped in the moderate and heavy S. mansoni parasitic load patients as compared with the control group and light S. mansoni parasite density patients. However, the mean of AST and ALT progressively elevated as the burden of S. mansoni increased. Conclusion. Most hematological and biochemical profiles were significantly lower in the Schistosoma mansoni-positive group as compared with the control group. Most hematological and biochemical profiles decline significantly as the parasite density increased. Hence, with Schistosoma treatment, supportive treatment against hematological and biochemical disorders is recommended.

Prevalence and Reinfection Rates of Schistosoma mansoni and Praziquantel Efficacy against the Parasite among Primary School Children in Sanja Town, Northwest Ethiopia.

BACKGROUND: Schistosomiasis is among the most widespread chronic infections in the world. The magnitude of the infection may show variations across different areas with respect to time. Praziquantel is a first line drug of choice for the treatment of schistosomiasis although its low cure rate has been reported in different parts of the world. Thus, an assessment of the magnitude of the diseases, the efficacy of currently available drugs, and reinfection rates is crucial. OBJECTIVE: Our principal objective is to determine the prevalence and reinfection rates of Schistosoma mansoni and to evaluate the efficacy of PZQ against Schistosoma mansoni. METHOD: A school-based cross-sectional study was conducted on Sanja Elementary Schools, Sanja town, northwest Ethiopia. Stool specimens were examined using Kato-Katz method. Schoolchildren who tested positive for intestinal schistosomiasis and fulfilled the inclusion criteria took part in the efficacy and reinfection study. Positive participants were treated with 40 mg/kg of Praziquantel. Cure and egg reduction rates were evaluated three weeks after treatment. The intensity of infection was determined following the WHO's guideline. Moreover, the reinfection rate of those who were cured was evaluated after a six-month posttreatment period. Data were analyzed using SPSS version 20. RESULTS: At baseline, 130 (35%) of the 372 schoolchildren were found infected with Schistosoma mansoni. Out of the 130 infected schoolchildren, 112 (86.2%) had moderate infection intensity. Among the S. mansoni positive schoolchildren, 80 were included as study participants for the evaluation of PZQ efficacy, based on the inclusion criteria established by WHO. The cure and egg reduction rates were found to be 90% (72/80) and 99.5%, respectively. Of the seventy-two schoolchildren considered for the determination of reinfection rate, after 6 months of posttreatment, 13.9% were found to be reinfected. CONCLUSION: The schoolchildren in the three primary schools of Sanja are at moderate risk of the infection caused by S. mansoni. Although the therapeutic potency of PZQ at 40 mg/kg was efficient against S. mansoni, a high rate of reinfection was reported in the study site, suggesting the need for integrated schistosomiasis control measures.

Prevalence of intestinal helminth infection among school children in Maksegnit and Enfranz Towns, northwestern Ethiopia, with emphasis on Schistosoma mansoni infection.

BACKGROUND: Schistosomiasis is endemic in Ethiopia and previously unknown transmission foci have been reported from time to time in different parts of the country. Further surveys are required in areas where endemicity of the disease is not known to cover them with control program if transmission is taking place. This study, therefore, aims to assess the magnitude of schistosomiasis mansoni and soil-transmitted helminthiasis in Maksegnit and Enfranz Towns, northwestern Ethiopia. METHODS: Cross-sectional parasitological and malacological surveys were conducted in three schools found in Maksegnit and Enfranz Towns. Stool specimens were collected from 550 randomly selected school children (age range 5 to 17 years) and processed for microscopic examination using Kato-Katz method (single smear per stool sample). Malacological survey was conducted in Gumara and Garno Rivers found in the study areas. Biomphalaria pfeifferi snails collected from the two rivers were individually exposed to artificial light in order to induce cercarial shedding. Laboratory-bred Swiss albino mice were exposed to the cercariae and definite identification of the schistosome species was made based on morphology. RESULTS: The overall prevalence of S. mansoni infection was found to be 49%; however, it varied by schools, with Selam having 60.7%, and Maksegnit Number 1 and 2 having 45.8 and 39.6%, respectively. The respective mean intensity of S. mansoni infection among school children in Selam, Maksegnit Number 1 and Maksegnit Number 2 Schools were 243, 194 and 183 eggs per gram of stool (epg). In all the study areas there was no difference in prevalence of S. mansoni infection in relation to age, however, the prevalence varied by sex, with males having highest prevalence (54.5% vs 44.1%) (p = 0.012). Adult S. mansoni worms were harvested from mice exposed to cercariae shed from B. pfeifferi on the 6(th) week post-exposure. The prevalence of Ascaris lumbricoides single infection was 16.5% while its co-infection with S. mansoni was 18.2%. CONCLUSION: Infections of young children, findings of schistosome infected snails, establishment of mice infection and harvesting adult worms from the lab-bred mice confirm that autochthonous transmission is taking place in the study areas. Hence, preventive chemotherapy with praziquantel should be put in place, complemented with other measures such as provision of sanitary facilities and health education, to control morbidity and transmission of schistosomiasis and soil-transmitted helminthiasis in the study areas.

Status of Schistosoma mansoni prevalence and intensity of infection in geographically apart endemic localities of Ethiopia: a comparison.

BACKGROUND: Schistosomiasis is one of the chronic and neglected tropical diseases affecting rural communities. Heavy infections contribute to anemia and can retard children's growth, physical activity and cognitive function. This study was conducted in order to determine the prevalence, intensity and variation of Schistosoma mansoni infection among human subjects in geographically apart localities. The prevalence and intensity of Schistosoma mansoni infection was assessed in three geographically apart endemic areas of Ethiopia from May to August 2010 through cross-sectional approach. METHODS: A total of 1073 individuals (528 males and 545 females) were found eligible for the study. Their age ranged from 5-60 years with mean age of 11 years in Wondo Genet, 22 years in Kemissie and 24 years in Sille-Elgo. Small plastic sheets were distributed to the study participants and sizable stool specimens were collected and examined using Kato-Katz method (41.7mg template). RESULT: The prevalence of Schistosoma mansoni infection among the study participants in Kemissie, Wondo Genet and Sille-Elgo was 89.6%, 59.9%, and 31.6%, respectively. The highest geometric mean of egg per gram of stool for Kemissie, Wondo Genet and Sille-Elgo was, 5208 and 346, 8472 and 252, 3960 and 91, respectively. CONCLUSION: It was observed that there was comparable variation in the prevalence and intensity of infection among the study localities. Moreover, it is indicated that S. mansoni is still an ongoing public health problem which requires integrated intervention activities in the country.

Epidemiological study on Schistosoma mansoni infection in Sanja area, Amhara region, Ethiopia.

BACKGROUND: The epidemiology of schistosomiasis is well documented and its geographic distribution has been mapped and there is an ongoing mapping in Ethiopia. Nevertheless, new transmission foci have been discovered in different parts of the country. The objective of this study was to assess the establishment of transmission and determine the prevalence of Schistosoma mansoni infection in school children from Sanja Town, northwest Ethiopia. METHODS: A cross-sectional parasitological survey involving 384 school children in two primary schools of Sanja Town was conducted between February and April 2013. Stool specimens were collected and microscopically examined using Kato-Katz and Sodium acetate-acetic acid-formalin (SAF) concentration methods. Malacological survey was also carried out to identify snail intermediate hosts and larval infection rate in the snail. The snails collected were checked for trematode infection by shedding. Observation was also made on water contact habits of the study population. RESULTS: The prevalence of Schistosoma mansoni infection using Kato-Katz method was high among male (79.5%) children in Sanja Primary school while it was high among female (75%) children in Ewket Amba Primary school. The prevalence of Schistosoma mansoni infection among Sanja Primary school children in the age groups 5-9 and 10-14 years were 84.6% and 75.2%, respectively while in Ewket Amba Primary school, the prevalence was 66% and 77.9% in the age groups 5-9 and 10-14 years respectively. The prevalence of schistosome infection in Biomphalaria pfeifferi was 16.9% and 0.027% during February and April, respectively. S. mansoni infection was successfully established in laboratory mice and adult worms were harvested after six weeks of laboratory maintenance. Observations made on water contact activities showed swimming, bathing and washing in the river and the stream as the high risk activities for Schistosoma mansoni infection. CONCLUSION: The study has shown establishment of transmission of schistosomiasis mansoni in Sanja Town. Therefore, appropriate integrated control measures need to be introduced to reduce morbidity in the population and also to control the transmission of schistosomiasis in the study area.