High pyrethroid-resistance intensity in Culex quinquefasciatus (Say) (Diptera: Culicidae) populations from Jigawa, North-West, Nigeria.

This study examined pyrethroid resistance intensity and mechanisms in Culex quinquefasciatus (Say) (Diptera: Culicidae) populations from Jigawa, North-West Nigeria. Resistance statuses to permethrin, lambda-cyhalothrin and alphacypermethrin were determined with both WHO and CDC resistance bioassays. Synergist assay was conducted by pre-exposing the populations to Piperonyl butoxide (PBO) using the WHO method. Resistance intensities to 2x, 5x and 10x of diagnostic concentrations were determined with the CDC bottle method. Species analysis and presence of knockdown mutation (Leu-Phe) were done using Polymerase Chain Reaction (PCR). Results showed that Cx. quinquefasciatus was the only Culex spp. present and "Kdr-west" mutation was not detected in all analyzed samples. Using WHO method, Cx. quinquefasciatus resistance to permethrin was detected in Dutse (12.2%) and Kafin-Hausa (77.78%). Lambda-cyhalothrin resistance was recorded only in Kafin-Hausa (83.95%) with resistance suspected in Ringim (90%). Resistance to alphacypermethrin was recorded in all locations. Pre-exposure to PBO led to 100% mortality to alphacypermethrin and lambda-cyhalothrin in Ringim while mortality to permethrin and alphacypermethrin in Dutse increased from 12.2% to 97.5% and 64.37% to 79.52% respectively. Using CDC bottle bioassay, resistance was also recorded in all populations and the result shows a significant positive correlation (R2 = 0.728, p = 0.026) with the result from the WHO bioassay. Results of resistance intensity revealed a very high level of resistance in Kafin-Hausa with susceptibility to lambda-cyhalothrin and alphacypermethrin not achieved at 10x of diagnostic doses. Resistance intensity was also high in Dutse with susceptibility to all insecticides not achieved at 5x of diagnostic doses. Widespread and high intensity of resistance in Cx. quinquefasciatus from North-West Nigeria is a major threat to the control of diseases transmitted by Culex and other mosquito species. It is a challenge that needs to be adequately addressed so as to prevent the failure of pyrethroid-based vector control tools.

A holistic approach is needed to control the perpetual burden of soil-transmitted helminth infections among indigenous schoolchildren in Malaysia.

A cross-sectional survey was conducted among 1,142 Orang Ali schoolchildren in six states of Peninsular Malaysia to investigate the current prevalence and risk factors of STH infections. Faecal samples were examined using direct smear, formalin-ether sedimentation, Kato-Katz, and Harada-Mori methods. A pre-tested questionnaire was used to collect information on the demographic, socioeconomic, personal hygiene, and health status of the participants. Overall, 70.1% (95% CI = 67.4, 72.7) of the participants were infected with at least one of the STH species. The prevalence of Ascaris lumbricoides, Trichuris trichiura, and hookworm infections was 63.1%, 61.8% and 11.5%, respectively. Moderate-to-heavy STH infections accounted for 61.3% of the total infections. Univariate and logistic regression analyses revealed different sets of risk factors, with age (> 10 years) being the significant risk factor of all three STH species. Moreover, other species-specific risk factors were identified including being a member of the Senoi tribe, family size (≥ 7 members), school size (150-250 pupils), maternal unemployment, unimproved source of drinking water, lacking improved toilet in the house, inadequate WASH facilities at school, not washing hands before eating, and not washing fruits before eating; presence of domestic animals, and not wearing shoes when outside. The high prevalence of STH infections found in the study population exceeds the WHO policy intervention threshold (20% prevalence). Thus, an innovative holistic approach should be adopted to control STH infections among these children as part of the efforts to improve the quality of life of the entire Orang Asli population. .

Prevalence and risk factors of Strongyloides stercoralis infection among Orang Asli schoolchildren: new insights into the epidemiology, transmission and diagnosis of strongyloidiasis in Malaysia.

This cross-sectional study aimed to determine the prevalence and risk factors of S. stercoralis infection among 1142 Orang Asli primary schoolchildren in six different states of Peninsular Malaysia. Fecal samples were examined using direct smear, formalin-ether sedimentation (FES), agar plate culture (APC) and PCR techniques. Overall, 15.8% of the children were found to be infected with S. stercoralis. The prevalence was 0.2, 1.3, 15.2 and 13.7% by direct smear, FES, APC and PCR, respectively. Multivariate analysis showed that an age of >10 years, being male, belonging to a Proto-Malay tribe, belonging to the Senoi tribe, indiscriminate defecation, using an unimproved water source for drinking water and not wearing shoes when outside were the significant risk factors of infection among these children. In conclusion, we provide new evidence on the occurrence of S. stercoralis in Malaysia to show that there is a relatively high prevalence of infection among Orang Asli schoolchildren. Therefore, the use of specific methods for detecting S. stercoralis should be considered when screening these children for intestinal parasites. Moreover, prevention and control measures specific to S. stercoralis should be integrated into the intestinal parasitic infections control programme in Malaysia.

The burden and epidemiology of polyparasitism among rural communities in Kano State, Nigeria.

BACKGROUND: Parasitic infections constitute a major public health problem worldwide, particularly among underprivileged communities in developing countries including Nigeria. The present study aimed to determine the epidemiology of polyparasitism (multiple parasitic infections) among rural communities in Kano State, North Central Nigeria. METHODS: A total of 551 individuals were screened for the presence of intestinal, urogenital and blood parasites by using different diagnostic techniques. Demographic, socioeconomic, household and behavioural characteristics were collected using a pre-tested questionnaire. RESULTS: Overall, 84.0% (463/551) of the participants were found to be infected with at least one parasite species, with 51.2% (282/551) of them having polyparasitism. The most prevalent parasites were Plasmodium falciparum (60.6%) followed by Blastocystis sp. (29.2%) and hookworm (15.4%). No significant association was found between malaria and helminth infections (p>0.05). Univariate and multivariate analyses showed that the presence of other family members who had intestinal polyparasitism (adjusted odds ratio [AOR]=4.12; 95% CI=2.72, 6.24), walking barefoot outside (AOR=1.70; 95% CI=1.09, 2.63) and being male (AOR=1.74; 95% CI=1.14, 2.66) were the significant risk factors of intestinal polyparasitism among the population studied. CONCLUSION: Polyparasitism is highly prevalent among rural communities in Kano State. Therefore, effective, sustainable and integrated control measures should be identified and implemented to significantly reduce the burden and consequences of these infections in rural Nigeria.

Occurrence of Acanthamoeba genotypes in Central West Malaysian environments.

Acanthamoeba species are ubiquitous free-living protozoa that can be found worldwide. Occasionally, it can become parasitic and the causative agent of acanthamoebic keratitis (AK) and Granulomatous Amoebic Encephalitis (GAE) in man. A total of 160 environmental samples and 225 naturally-infected animal corneal swabs were collected for Acanthamoeba cultivation. Acanthamoeba was found to be high in samples collected from environments (85%, 136/160) compared to infected animal corneas (24.89%, 56/225) by microscopic examination. Analysis of nucleotide sequence of 18S rRNA gene of all the 192 cultivable Acanthamoeba isolates revealed 4 genotypes (T3, T4. T5 and T15) with T4 as the most prevalent (69.27%, 133/192) followed by T5 (20.31%), T15 (9.90%) and T3 (0.52%). Genotype T4 was from the strain of A. castellanii U07401 (44.27%), A. castellanii U07409 (20.83%) and A. polyphagaAY026243 (4.17%), but interestingly, only A. castellanii U07401 was detected in naturally infected corneal samples. In environmental samples, T4 was commonly detected in all samples including dry soil, dust, wet debris, wet soil and water. Among the T4, A. castellanii (U07409) strains were detected high occurrence in dry (45%) followed by aquatic (32.50%) and moist (22.50%) samples but however A. castellanii (U07401) strains were dominant in dry samples of soil and dust (93.10%). Subsequently, genotype T5 of A. lenticulata (U94741) strains were dominant in samples collected from aquatic environments (58.97%). In summary, A. castellanii (U07401) strains were found dominant in both environmental and corneal swab samples. Therefore, these strains are possibly the most virulent and dry soil or dusts are the most possible source of Acanthamoeba infection in cats and dogs corneas.

First molecular epidemiology of Entamoeba histolytica, E. dispar and E. moshkovskii infections in Yemen: different species-specific associated risk factors.

OBJECTIVES: To investigate the molecular epidemiology of Entamoeba histolytica, E. dispar and E. moshkovskii infections among rural communities in Yemen. METHODS: In a community-based study, faecal samples were collected from 605 participants and examined by wet mount, formalin-ether sedimentation, trichrome staining and nested multiplex PCR techniques. Demographic, socio-economic and environmental information was collected using a pre-tested questionnaire. RESULTS: Overall, 324 (53.6%) of the samples were positive for Entamoeba cysts and/or trophozoites by microscopic examination. Molecular analysis revealed that 20.2%, 15.7% and 18.2% of the samples were positive for E. histolytica, E. dispar and E. moshkovskii, respectively. Multivariate analysis showed different sets of species-specific risk factors among these communities. Educational level was identified as the significant risk factor for E. histolytica; age and gender were the significant risk factors for E. moshkovskii; and sources of drinking water and consumption of unwashed vegetables were the significant risk factors for E. dispar. Moreover, living in coastal/foothill areas and presence of other infected family members were risk factors for both E. histolytica and E. moshkovskii infections. CONCLUSION: The study reveals that Entamoeba spp. infection is highly prevalent among rural communities in Yemen, with E. histolytica, E. dispar and E. moshkovskii differentiated for the first time. Identifying and treating infected family members, providing health education pertinent to good personal and food hygiene practices and providing clean drinking water should be considered in developing a strategy to control intestinal parasitic infections in these communities, particularly in the coastal/foothill areas of the country.

Different patterns of pfcrt and pfmdr1 polymorphism in Plasmodium falciparum isolates from Tehama region, Yemen.

Introduction. Despite the efforts of the malaria control programme, malaria morbidity is still a common health problem in Yemen, with 60% of the population at risk. Plasmodium falciparum is responsible for 99% of malaria cases. The emergence in Yemen of parasite resistance to chloroquine (CQ) prompted the adoption of artemisinin combination therapy (ACT) in 2009, which involves the use of artesunate plus sulphadoxine-pyrimethamine (AS + SP). However, CQ was retained as the drug of choice for vivax malaria. To assess the impact of the change in the malaria treatment policy five years after its introduction, the present study investigated the mutations in the CQ resistance transporter (pfcrt) and multidrug resistance 1 (pfmdr1) genes. Method. A molecular investigation of 10 codons of pfcrt (72-76, 220, 271, 326, 356, and 371) and five codons of pfmdr1 (86, 184, 1034, 1042, and 1246) was conducted on P. falciparum isolates from districts with the highest malaria endemicity in the Hodeidah and Al-Mahwit governorates in Tehama region, Yemen. A total of 86 positive cases of falciparum monoinfection were investigated for the presence of mutations related to CQ and other antimalarials using a PCR-RFLP assay. Results. There was a wide prevalence of pfcrt gene mutations with the pfcrt 76T CQ resistance marker being predominant (97.7%). The prevalence of other pfcrt mutations varied from high (75E: 88%) to moderate (74I: 79.1%, 220S: 69.8%, 271E and 371I: 53.5%) or low (326S: 36%, 72S: 10.5%). Mutated pfcrt 72-76 amino acids haplotypes were highly prevalent (98.8%). Among these, the CVIET classic, old-world African/Southeast Asian haplotype was the most predominant, and was mostly found in the isolates from the Khamis Bani Saad district of Al-Mahwit (93.1%) and the AdDahi district of Hodeidah (88.9%). However, it was only found in 26.3% of the isolates from the Bajil district of Hodeidah. Surprisingly, the SVMNT new-world South American haplotype was exclusively detected in 9.3% of the isolates from the Bajil district of Hodeidah. Mutations at Y184F of pfmdr1 were found in all isolates (100%) from all districts. The mutation for codons 1034C and 86Y were found only in the isolates from the AdDahi and Khamis Bani Saad districts. Overall, the AdDahi and Khamis Bani Saad districts were similar in terms of carrying most of the mutations in the pfcrt and pfmdr1 genes, while there was a lower prevalence of mutation in the isolates from the Bajil district. Conclusion. The high prevalence of mutations in pfcrt 5 years after the official cessation of CQ use against P. falciparum suggests that there is sustained CQ pressure on P. falciparum isolates in the study area. Moreover, the low prevalence of mutations in the pfmdr1 gene could be a good indicator of the high susceptibility of P. falciparum isolates to antimalarials other than CQ. A new strategy to ensure the complete nationwide withdrawal of CQ from the private drug market is recommended.

A tale of two communities: intestinal polyparasitism among Orang Asli and Malay communities in rural Terengganu, Malaysia.

BACKGROUND: Intestinal parasitic infections (IPIs) are still major health problems in many developing countries including Malaysia, particularly in the poor and socioeconomically deprived rural and remote communities in Peninsular Malaysia. This study was conducted to determine the prevalence of IPIs and to identify the key factors associated with intestinal polyparasitism as well as to evaluate the knowledge, attitude and practices (KAP) on IPIs among rural Orang Asli and Malay communities in Terengganu, Malaysia. METHODS: A cross-sectional study was conducted among 340 participants (165 Orang Asli and 175 Malay) aged ≤ 15 years from the Hulu Terengganu and Kemaman districts of Terengganu. Faecal samples were examined for the presence of intestinal parasites by using direct smear, formalin-ether sedimentation, trichrome stain, modified Ziehl Neelsen stain, in vitro cultivation in Jones' medium, Kato Katz and Harada Mori techniques. Demographic, socioeconomic, environmental and behavioural information of the participants and their KAP for IPIs were collected by using a pre-tested questionnaire. RESULTS: Overall, 149 (90.3 %) Orang Asli and 43 (24.6 %) Malay children were infected by at least one parasite species. The overall prevalences of intestinal polyparasitism among the Orang Asli and Malay were 68.5 % (113/165) and 14.3 % (25/175), respectively. Multiple logistic regression analysis showed that using unsafe water supply as a source for drinking water, the presence of domestic animals, not wearing shoes when outside, not washing vegetables before consumption, not washing hands after playing with soil, indiscriminate defecation and the low level of mother's education were the key risk factors for intestinal polyparasitism among the Orang Asli, while working mothers and the presence of domestic animals were the risk factors among the Malay children. Almost all the Malays were well aware about the IPIs while Orang Asli respondents had a poor level of related awareness. CONCLUSIONS: This study demonstrates that IPIs are highly prevalent in rural Terengganu, Malaysia. Community awareness about IPIs was found to be imperative in protecting Malay children from these infections. An integrated control programme for the prevention and control of IPIs is highly recommended for these communities, with a special emphasis on the Orang Asli population.

PREVALENCE AND RISK FACTORS OF SCHISTOSOMIASIS AMONG HAUSA COMMUNITIES IN KANO STATE, NIGERIA.

Schistosomiasis remains one of the most prevalent neglected tropical diseases especially in Nigeria which has the greatest number of infected people worldwide. A cross-sectional study was conducted among 551 participants from Kano State, North Central Nigeria. Fecal samples were examined for the presence of Schistosoma mansoni eggs using the formalin-ether sedimentation method while the urine samples were examined using the filtration technique for the presence of S. haematobium eggs. Demographic, socioeconomic and environmental information was collected using a pre-validated questionnaire. The overall prevalence of schistosomiasis was 17.8%, with 8.9% and 8.3% infected with S. mansoni and S. haematobium, respectively and 0.5% presenting co-infection with both species. The multiple logistic regression analysis revealed that age < 18 years (OR = 2.13; 95% CI; 1.34- 3.41), presence of infected family members (OR = 3.98; 95% CI; 2.13-7.46), and history of infection (OR = 2.87; 95% CI; 1.87- 4.56) were the significant risk factors associated with schistosomiasis in these communities. In conclusion, this study revealed that schistosomiasis is still prevalent among Hausa communities in Nigeria. Mass drug administration, health education and community mobilization are imperative strategies to significantly reduce the prevalence and morbidity of schistosomiasis in these communities.

Is Nigeria winning the battle against malaria? Prevalence, risk factors and KAP assessment among Hausa communities in Kano State.

BACKGROUND: Malaria is one of the most severe global public health problems worldwide, particularly in Africa, where Nigeria has the greatest number of malaria cases. This community-based study was designed to investigate the prevalence and risk factors of malaria and to evaluate the knowledge, attitudes, and practices (KAP) regarding malaria among rural Hausa communities in Kano State, Nigeria. METHODS: A cross-sectional community-based study was conducted on 551 participants from five local government areas in Kano State. Blood samples were collected and examined for the presence of Plasmodium species by rapid diagnostic test (RDT), Giemsa-stained thin and thick blood films, and PCR. Moreover, demographic, socioeconomic, and environmental information as well as KAP data were collected using a pre-tested questionnaire. RESULTS: A total of 334 (60.6 %) participants were found positive for Plasmodium falciparum. The prevalence differed significantly by age group (p < 0.01), but not by gender or location. A multivariate analysis showed that malaria was associated significantly with being aged 12 years or older, having a low household family income, not using insecticide treated nets (ITNs), and having no toilets in the house. Overall, 95.6 % of the respondents had prior knowledge about malaria, and 79.7, 87.6 and 95.7 % of them knew about the transmission, symptoms, and prevention of malaria, respectively. The majority (93.4 %) of the respondents considered malaria a serious disease. Although 79.5 % of the respondents had at least one ITN in their household, utilization rate of ITNs was 49.5 %. Significant associations between the respondents' knowledge concerning malaria and their age, gender, education, and household monthly income were reported. CONCLUSIONS: Malaria is still highly prevalent among rural Hausa communities in Nigeria. Despite high levels of knowledge and attitudes in the study area, significant gaps persist in appropriate preventive practices, particularly the use of ITNs. Innovative and Integrated control measures to reduce the burden of malaria should be identified and implemented in these communities. Community mobilization and health education regarding the importance of using ITNs to prevent malaria and save lives should be considered.

The Menace of Schistosomiasis in Nigeria: Knowledge, Attitude, and Practices Regarding Schistosomiasis among Rural Communities in Kano State.

BACKGROUND: Schistosomiasis is one of the most common neglected tropical diseases, especially in the developing countries in Africa, Asia and South America, with Nigeria having the greatest number of cases of schistosomiasis worldwide. This community-based study aims to evaluate the knowledge, attitude and practices (KAP) regarding schistosomiasis among rural Hausa communities in Kano State, Nigeria. METHODS: A cross-sectional study was carried out among 551 participants from Hausa communities in five local government areas in Kano State, North Central Nigeria. Demographic, socioeconomic and environmental information as well as KAP data were collected using a pre-tested questionnaire. Moreover, faecal and urine samples were collected and examined for the presence of Schistosoma mansoni and S. haematobium eggs respectively. RESULTS: The overall prevalence of schistosomiasis was 17.8%, with 8.9% and 8.3% infected with S. mansoni and S. haematobium respectively, and 0.5% had co-infection of both species. Moreover, 74.5% of the participants had prior knowledge about schistosomiasis with 67.0% of them how it is transmitted and 63.8% having no idea about the preventive measures. Three-quarters of the respondents considered schistosomiasis a serious disease while their practices to prevent infections were still inadequate, with only 34.7% of them seeking treatment from clinics/hospitals. Significant associations between the KAP and age, gender, education and employment status were reported. Multiple logistic regression analysis revealed that age, gender, history of infection and educational level of the respondents were the most important factors significantly associated with the KAP on schistosomiasis among this population. CONCLUSIONS: Schistosomiasis is still prevalent among Hausa communities in Nigeria and participants' knowledge about the disease was poor. Mass drug administration, community mobilization and health education regarding the cause, transmission and prevention of schistosomiasis and education about good personal and sanitary hygiene practices should be considered in order to significantly reduce the prevalence and morbidity of infection within these communities.

Knowledge, attitude, and practices towards schistosomiasis among rural population in Yemen.

BACKGROUND: Schistosomiasis is highly prevalent in Yemen, with an estimated 3 million cases, particularly among rural communities. This community-based study aims to evaluate the knowledge, attitude and practices (KAP) on schistosomiasis among rural communities in Yemen. METHODS: A cross-sectional study was carried out among 250 households from ten rural districts in Yemen. Overall, 400 children were screened for urogenital and intestinal schistosomiasis. Moreover, parents were interviewed using a pre-tested questionnaire to collect information about the demographic and socioeconomic information and their KAP concerning schistosomiasis. RESULTS: A total of 127 (31.8%) children were found to be excreting schistosome eggs in either their urine or faeces (22.5% S. haematobium and 8.0% S. mansoni). Although 92.4% of the respondents had heard about schistosomiasis, 49.8%, 68.0% and 47.2% had knowledge concerning the transmission, signs and symptoms, and prevention, respectively. In addition, 77.1% considered schistosomiasis as harmful while 48.5% believed that schistosomiasis could be prevented, albeit their practices to prevent infections were still inadequate. Significant associations between the KAP and age, education, employment status and household monthly income were reported (P < 0.05). Moreover, a significantly higher level of knowledge was reported among the respondents who had infected children compared to those with no infected family members (P < 0.05). Multiple logistic regression analysis revealed that the level of education and the history of schistosomiasis were the most important factors associated with the KAP concerning schistosomiasis among this population. CONCLUSION: This study reveals that knowledge about the cause, transmission, symptoms and prevention of schistosomiasis among the rural population in Yemen was inadequate, and that this could be a challenging obstacle to the elimination of schistosomiasis in these communities. Besides the current mass drug administration, school and community-based health education regarding schistosomiasis is imperative among these communities to significantly reduce the transmission and morbidity of schistosomiasis.

Detection of Schistosoma mansoni and Schistosoma haematobium by Real-Time PCR with High Resolution Melting Analysis.

The present study describes a real-time PCR approach with high resolution melting-curve (HRM) assay developed for the detection and differentiation of Schistosoma mansoni and S. haematobium in fecal and urine samples collected from rural Yemen. The samples were screened by microscopy and PCR for the Schistosoma species infection. A pair of degenerate primers were designed targeting partial regions in the cytochrome oxidase subunit I (cox1) gene of S. mansoni and S. haematobium using real-time PCR-HRM assay. The overall prevalence of schistosomiasis was 31.8%; 23.8% of the participants were infected with S. haematobium and 9.3% were infected with S. mansoni. With regards to the intensity of infections, 22.1% and 77.9% of S. haematobium infections were of heavy and light intensities, respectively. Likewise, 8.1%, 40.5% and 51.4% of S. mansoni infections were of heavy, moderate and light intensities, respectively. The melting points were distinctive for S. mansoni and S. haematobium, categorized by peaks of 76.49 ± 0.25 °C and 75.43 ± 0.26 °C, respectively. HRM analysis showed high detection capability through the amplification of Schistosoma DNA with as low as 0.0001 ng/µL. Significant negative correlations were reported between the real-time PCR-HRM cycle threshold (Ct) values and microscopic egg counts for both S. mansoni in stool and S. haematobium in urine (p < 0.01). In conclusion, this closed-tube HRM protocol provides a potentially powerful screening molecular tool for the detection of S. mansoni and S. haematobium. It is a simple, rapid, accurate, and cost-effective method. Hence, this method is a good alternative approach to probe-based PCR assays.

Genetic variation of pfhrp2 in Plasmodium falciparum isolates from Yemen and the performance of HRP2-based malaria rapid diagnostic test.

BACKGROUND: The genetic variation in the Plasmodium falciparum histidine-rich protein 2 (pfhrp2) gene that may compromise the use of pfhrp2-based rapid diagnostic tests (RDTs) for the diagnosis of malaria was assessed in P. falciparum isolates from Yemen. METHODS: This study was conducted in Hodeidah and Al-Mahwit governorates, Yemen. A total of 622 individuals with fever were examined for malaria by CareStart malaria HRP2-RDT and Giemsa-stained thin and thick blood films. The Pfhrp2 gene was amplified and sequenced from 180 isolates, and subjected to amino acid repeat types analysis. RESULTS: A total of 188 (30.2%) participants were found positive for P. falciparum by the RDT. Overall, 12 different amino acid repeat types were identified in Yemeni isolates. Six repeat types were detected in all the isolates (100%) namely types 1, 2, 6, 7, 10 and 12 while types 9 and 11 were not detected in any of the isolates. Moreover, the sensitivity and specificity of the used PfHRP2-based RDTs were high (90.5% and 96.1%, respectively). CONCLUSION: The present study provides data on the genetic variation within the pfhrp2 gene, and its potential impact on the PfHRP2-based RDTs commonly used in Yemen. CareStart Malaria HRP2-based RDT showed high sensitivity and specificity in endemic areas of Yemen.