Antimalarial drug efficacy and resistance in malaria-endemic countries in HANMAT-PIAM_net countries of the Eastern Mediterranean Region 2016-2020: Clinical and genetic studies.

INTRODUCTION: The World Health Organization recommends regular monitoring of the efficacy of nationally recommended antimalarial drugs. We present the results of studies on the efficacy of recommended antimalarials and molecular markers of artemisinin and partner resistance in Afghanistan, Pakistan, Somalia, Sudan and Yemen. METHODS: Single-arm prospective studies were conducted to evaluate the efficacy of artesunate-sulfadoxine-pyrimethamine (ASSP) in Afghanistan and Pakistan, artemether-lumefantrine (AL) in all countries, or dihydroartemisinin-piperaquine (DP) in Sudan for the treatment of Plasmodium falciparum. The efficacy of chloroquine (CQ) and AL for the treatment of Plasmodium vivax was evaluated in Afghanistan and Somalia, respectively. Patients were treated and monitored for 28 (CQ, ASSP and AL) or 42 (DP) days. Polymerase chain reaction (PCR)-corrected cure rate and parasite positivity rate at Day 3 were estimated. Mutations in the P. falciparum kelch 13 (Pfk13) gene and amplifications of plasmepsin (Pfpm2) and multidrug resistance-1 (Pfmdr-1) genes were also studied. RESULTS: A total of 1680 (249 for ASSP, 1079 for AL and 352 for DP) falciparum cases were successfully assessed. A PCR-adjusted ASSP cure rate of 100% was observed in Afghanistan and Pakistan. For AL, the cure rate was 100% in all but four sites in Sudan, where cure rates ranged from 92.1% to 98.8%. All but one patient were parasite-free at Day 3. For P. vivax, cure rates were 98.2% for CQ and 100% for AL. None of the samples from Afghanistan, Pakistan and Yemen had a Pfk13 mutation known to be associated with artemisinin resistance. In Sudan, the validated Pfk13 R622I mutation accounted for 53.8% (14/26) of the detected non-synonymous Pfk13 mutations, most of which were repeatedly detected in Gadaref. A prevalence of 2.7% and 9.3% of Pfmdr1 amplification was observed in Pakistan and Yemen, respectively. CONCLUSION: High efficacy of ASSP, AL and DP in the treatment of uncomplicated falciparum infection and of CQ and AL in the treatment of P. vivax was observed in the respective countries. The repeated detection of a relatively high rate of Pfk13 R622I mutation in Sudan underscores the need for close monitoring of the efficacy of recommended ACTs, parasite clearance rates and Pfk13 mutations in Sudan and beyond. Registration numbers of the trials: ACTRN12622000944730 and ACTRN12622000873729 for Afghanistan, ACTRN12620000426987 and ACTRN12617001025325 for Pakistan, ACTRN12618001224213 for Somalia, ACTRN12617000276358, ACTRN12622000930785 and ACTRN12618001800213 for Sudan and ACTRN12617000283370 for Yemen.

Status of insecticide resistance and its biochemical and molecular mechanisms in Anopheles stephensi (Diptera: Culicidae) from Afghanistan.

BACKGROUND: Insecticide resistance of Anopheles stephensi, the main malaria vector in eastern Afghanistan, has been reported previously. This study describes the biochemical and molecular mechanisms of resistance to facilitate effective vector control and insecticide resistance management. METHODS: Mosquito larvae were collected from the provinces of Kunar, Laghman and Nangarhar from 2014 to 2017. The susceptibility of the reared 3-4 days old adults was tested with deltamethrin 0.05%, bendiocarb 0.1%, malathion 5%, permethrin 0.75% and DDT 4%. Cytochrome P450 content and general esterase, glutathione S-transferase (GST) and acetylcholinesterase (AChE) activities were measured in the three field populations and the results were compared with those of the laboratory susceptible An. stephensi Beech strain. Two separate allele-specific PCR assays were used to identify L1014, L1014F and L1014S mutations in the voltage gated sodium channel gene of An. stephensi. Probit analysis, ANOVA and Hardy-Weinberg equilibrium were used to analyse bioassay, biochemical assay and gene frequency data respectively. RESULTS: The population of An. stephensi from Kunar was susceptible to bendiocarb, apart from this, all populations were resistant to all the other insecticides tested. The differences between all values for cytochrome P450s, general esterases, GSTs and AChE inhibition rates in the Kunar, Laghman and Nangarhar populations were statistically significant when compared to the Beech strain, excluding GST activities between Kunar and Beech due to the high standard deviation in Kunar. The three different sodium channel alleles [L1014 (wild type), L1014F (kdr west) and L1014S (kdr east)] were all segregated in the Afghan populations. The frequencies of kdr east mutation were 22.9%, 32.7% and 35% in Kunar, Laghman and Nangarhar populations respectively. Kdr west was at the lowest frequency of 4.44%. CONCLUSIONS: Resistance to different groups of insecticides in the field populations of An. stephensi from Kunar, Laghman and Nangarhar Provinces of Afghanistan is caused by a range of metabolic and site insensitivity mechanisms, including esterases, cytochrome P450s and GSTs combined with AChE and sodium channel target site insensitivity. The intensity and frequency of these mechanisms are increasing in these populations, calling for urgent reorientation of vector control programmes and implementation of insecticide resistance management strategies.

Prevalence of Trachoma in 72 Districts of Afghanistan in 2018-2019: Results of 35 Population-based Prevalence Surveys.

BACKGROUND: To determine where interventions are needed to eliminate trachoma as a public health problem, prevalence data are needed. We aimed to generate baseline population-based data on trachoma prevalence in suspected-endemic areas of Afghanistan. METHODS: Cross-sectional population-based prevalence surveys designed according to World Health Organization (WHO) recommendations were conducted in 35 evaluation units (EUs) covering 72 districts. In selected households, all resident individuals aged ≥1 year were examined for trachomatous inflammation-follicular (TF) and trachomatous trichiasis (TT) according to the WHO simplified trachoma grading system. Water, sanitation and hygiene access was assessed in households of survey participants. RESULTS: 104,104 people aged ≥1 year were examined, including 43,774 children aged 1-9 years and 46,439 people aged ≥15 years. The age-adjusted prevalence of TF in 1-9-year-olds was ≥5% in 3 EUs, with the highest EU TF prevalence being 7.8%. The age- and gender-adjusted prevalence of TT unknown to the health system in ≥15-year-olds was <0.2% in all EUs. The majority of households had access to an improved water source within 30 minutes of the house. However, only a minority of households had an improved latrine and/or a handwash station. CONCLUSIONS: Trachoma is not a public health problem in the majority of EUs surveyed. However, antibiotic mass drug administration, promotion of facial cleanliness and environmental improvement (the A, F and E components of the SAFE strategy) are needed for trachoma elimination purposes in three of the EUs surveyed in Afghanistan.

Elimination of morbidity due to soil-transmitted helminthiases among Afghan schoolchildren.

Large sectors of the Afghan population have limited access to safe water and sanitation, which increases the risk of transmission of water- and food- borne diseases, including Soil-Transmitted Helminth (STH) infections. STHs interfere with the human host's health status, and their burden of disease is highest among children of school age. Based on the results of a nationwide survey conducted in 2003, which showed an STH prevalence of 47.2%, and with the aim of reducing morbidity among school children, Afghanistan has been conducting nationwide deworming for preschool-age and school-age children since 2004. In 2017, 14 years after the first baseline assessment, a follow-up survey was carried out among schoolchildren aged 8-10 years to provide an update on STH epidemiology and facilitate evidence-informed planning of future deworming campaigns. Stool samples were collected from 2263 pupils aged 8-10 years in five provinces representing the different ecological zones of the country - Kabul, Balkh, Herat, Nangarhar and Kandahar. Microscopic examination was carried out by the Kato-Katz thick smear technique, to assess the presence and the number of parasites and/or their eggs. The survey revealed that 26.6% of the sample was infected with at least one of the STH, a marked decrease from the level registered in 2003. The most prevalent infection was the one with A. lumbricoides (25.7%), followed by T. trichiura (1.0%) and hookworms (0.1%). All positive children were noted to have light-intensity infections, compared to the previous survey where 9.7% of the sample had moderate-to-heavy intensity infections. Only 0.2% of the children had co-infection with two or more parasites. Meanwhile, 6.8% of the students were found infected with the dwarf tapeworm, Hymenolepis nana. The absence of infections of moderate-to-heavy intensity after several yearly rounds of deworming and overall improvements in provision of safe water and sanitation, indicates successful control of morbidity due to STH and, overall, their elimination as a public-health problem from Afghanistan. Nevertheless, current levels of prevalence of infection still show persistence of active transmission of STHs, thus justifying the continued implementation of mass deworming interventions among children. The permanent elimination of STH transmission, however, will be possible only when the country reaches a sanitation level sufficient to impede fecal contamination of the environment with human excreta.