Schistosoma mansoni and soil-transmitted helminths among schoolchildren in An-Nadirah District, Ibb Governorate, Yemen after a decade of preventive chemotherapy.

The Ministry of Public Health in Yemen continues the implementation of school and community-based preventive chemotherapy with praziquantel and albendazole for the control and elimination of schistosomiasis and soil-transmitted helminths (STH). The latest remapping to update the distribution of schistosomiasis and STH was conducted seven years ago. This study aimed to estimate the prevalence, intensity and associated risk factors of Schistosoma mansoni and STH among schoolchildren in An-Nadirah District, Ibb Governorate, Yemen. A cross-sectional study was carried out among schoolchildren aged 6-15 years in four selected schools. Biological, demographic, socioeconomic and environmental data were collected using a pre-tested questionnaire. S. mansoni and STH eggs were detected and counted by the microscopic examination of Kato-Katz fecal smears. Out of 417 schoolchildren, 17.0% were infected with at least one intestinal helminth. Prevalence of S. mansoni and STH were 6.5% and 9.1%, respectively. The most prevalent parasite among STH was Ascaris lumbricoides (8.4%). Unemployed fathers (Adjusted Odds Ratio (AOR) = 3.2; 95% Confidence interval (CI): 1.23, 8.52; P = 0.018), eating exposed food (AOR: 2.9; 95%CI = 1.24, 6.89; P = 0.014), not washing hands before eating and after defecation (AOR: 4.8; 95%CI = 1.77, 12.81; P = 0.002), and schools located close to water stream (AOR: 22.1; 95%CI = 5.12, 95.46; P <0.001) were independent risk factors of ascariasis. Swimming in ponds/stream (AOR: 3.9; 95%CI = 1.63, 9.55; P = 0.002), and schools close to the stream (AOR: 24.7; 95%CI = 3.05, 200.07; P = 0.003) were independent risk factors of intestinal schistosomiasis. The present study does not indicate a reduction in the prevalence of intestinal schistosomiasis in this rural area since the latest remapping conducted in 2014, although ascariasis was reduced by half. The prevalence of the two parasites was highly focal in areas close to the valley, suggesting a significant role of the stream in sustaining and accelerating the parasitic infection. Children practicing swimming and having poor hygienic practices were at high exposure to S. mansoni and A. lumbricoides, respectively. Water, Sanitation and Hygiene intervention, school-based health education, and snail control, in addition to mass drug administration, will help in the interruption of transmission of schistosomiasis and STH.

Prevalence of Intestinal Parasites among Immunocompromised Patients, Children, and Adults in Sana'a, Yemen.

Intestinal parasite infection (IPI) is still a very important public health issue. The severity of the parasitic disease has been reported as a high infection in immunocompromised patients and children. Hence, this study aimed to investigate the prevalence of intestinal parasites among immunocompromised patients and children with various gastrointestinal system complications in Sana'a city, Yemen, with different variables, including genus and age, and explore the risk factors associated with parasitic intestinal infections. The study socioeconomic data and certain behavioral and environmental risk factors and stool samples were collected from immunocompromised adult and children's patients, including children (one to eight years old), pregnant women, diabetes mellitus patients, cancer patients, HIV patients, and older adults. Out of 436 fecal samples, the overall prevalence rate of IPIs among immunocompromised patients and children in Sana'a was 51.8%. In contrast, the rate of infection in children (26.1%) was higher than that in old patients (25.7%) and in females (38.5%) and higher than that in males (13.3%). The protozoa (44.5%) have been shown more than intestinal helminths (7.3%) in samples, and the most common intestinal protozoan was Giardia lamblia and Entamoeba histolytica (13.8% and 12.8%), respectively. The most common intestinal helminthiasis was Hymenolepis nana with 1.8%. Concluding that the rate of infection was high for several reasons, including lack of commitment to hygiene as not handwashing after using the toilet (88.9%), eating uncovered food (56.3%), poor sanitation as lack of water sources (59.5%), reduced health education, and presence of other family members infected by parasites (61.3%). Interventions are required to reduce intestinal parasites, including health education on personal hygiene for patients, increasing awareness, and improving the environment and healthcare system.

National mapping of schistosomiasis, soil-transmitted helminthiasis and anaemia in Yemen: Towards better national control and elimination.

BACKGROUND: Schistosomiasis (SCH) and soil-transmitted helminthiasis (STH) are known to be endemic in Yemen. However, the distribution of both diseases had not previously been assessed by a well-structured national mapping study covering all governorates. The main aim of this study was, therefore, to map the prevalence of SCH and STH in Yemen in order to better inform implementation of effective national control and elimination interventions. The assessment of the distribution of anaemia was also included as a well-known consequence of infection with both SCH and STH. Secondarily, the study aimed to provide a broad indication of the impact of large-scale treatment on the distribution of infection. METHODOLOGY AND PRINCIPAL FINDINGS: To achive these aims, 80,432 children (10-14 years old) from 2,664 schools in 332 of Yemen's 333 districts were included, in 2014, into this national cross-sectional survey. Countrywide, 63.3% (210/332) and 75.6% (251/332) of districts were found to be endemic for SCH and STH respectively. More districts were affected by intestinal than urogenital SCH (54.2% and 31.6% respectively). SCH infection was mostly mild and moderate, with no districts reporting high infection. One quarter (24.4%) of Yemeni districts had high or moderate levels of Ascaris lumbricoides infection. Infection with Trichuris trichiura was the second most common STH (44.9% of districts infected) after A. lumbricoides (68.1%). Hookworm was the least prevalent STH (9.0%). Anaemia was prevalent in 96.4% of districts; it represented a severe public health problem (prevalence ≥ 40%) in 26.5% of districts, and a mild to moderate problem in two thirds of the districts (33.7% and 36.1% respectively). CONCLUSION: This study provided the first comprehensive mapping of SCH, STH, and anaemia across the country. This formed the basis for evaluating and continuing the national control and elimination programme for these neglected tropical diseases in Yemen.

Toxoplasma gondii Infection Among Institutionalized Children with Down syndrome in Sana'a city, Yemen: Implications of Low IgG Seroprevalence.

PURPOSE: To assess the IgG seroprevalence of Toxoplasma gondii as an indicator of past exposure and immunity against infection among children with Down syndrome (DS) in Sana'a city, Yemen. This preliminary study is justified by the primary immunodeficiency of children with DS and the opportunistic nature of the parasite, considering the vague situation of anti-Toxoplasma IgG seroprevalence among children with DS because of neglecting its study on local and global scales. METHODS: This descriptive, facility-based, cross-sectional study was conducted among 107 children with DS hosted in six randomly selected rehabilitation centers for children with special needs in Sana'a city. Demographics of children and their mothers' knowledge of toxoplasmosis were collected using a pre-designed, structured questionnaire. Anti-Toxoplasma IgG antibodies were measured in the sera of children using electrochemiluminescence assay. RESULTS: Of 107 children with DS, 3 (2.8%) were seropositive for anti-Toxoplasma IgG. Approximately two-thirds (71/106) of the mothers of children with DS were aware of toxoplasmosis. Of whom, 83.1% (59/71) were aware of its congenital complications. CONCLUSION: The majority of children with DS in Sana'a city are seronegative for anti-Toxoplasma IgG, where the seropositivity rate is lower than 3.0%. Therefore, children with DS are non-immune and susceptible to the acquisition of primary infections during their life. Further analytical studies are recommended to determine whether the defective immune response of children with DS is associated with false seronegativity, to assess the role of their mothers' knowledge in reducing their exposure to infection if they were confirmed truly seronegative and to identify the predictors of infection among them.

Intestinal schistosomiasis among schoolchildren in Sana'a Governorate, Yemen: Prevalence, associated factors and its effect on nutritional status and anemia.

Intestinal schistosomiasis is a neglected tropical disease, causing morbidity and mortality in tropical and subtropical countries. Despite the frequent implementation of mass drug administration with praziquantel, the reinfection with Schistosoma mansoni is still common in Yemen. In addition, there is a scarcity of information on the impact of S. mansoni on nutritional status and anemia among schoolchildren. The present study aimed to determine prevalence and risk factors of intestinal schistosomiasis and investigate its impact on nutritional status and anemia among schoolchildren in Sana'a Governorate, Yemen. It was conducted in 2018 on 445 schoolchildren aged 5-15 years. Biodata, socio-economic, demographic, behavioral and environmental data were collected using a standard questionnaire. S. mansoni was identified and quantified by microscopic examination of Kato-Katz fecal smear. Hemoglobin concentration and anthropometric measurements were estimated using standard methods. The prevalence of S. mansoni was higher in Al-Haimah Al-Dakheliah (33.9%) than Bani Mater (1.4%). Household without tap water (Adjusted Odds Ratio (AOR) = 2.9, 95% Confidence interval (CI): 1.12, 7.55, P = 0.028) was the independent risk factor of the infection. The prevalence of wasting and stunting was 25.0% (95%CI: 21.2%, 29.2%) and 45.8% (95%CI: 41.2%, 50.5%), respectively. The prevalence of underweight among schoolchildren aged 5-10 years was 27.3% (95%CI: 21.9%, 33.4%). The prevalence of anemia was 31.7% (95%CI: 27.5%, 36.2%) with 0.5%, 21.1% and 10.1% being severe, moderate and mild anemia, respectively. S. mansoni (AOR = 4.1, 95%CI: 2.16, 7.84, P < 0.001) and early adolescence (AOR = 6.8, 95%CI: 4.26, 10.82, P < 0.001) were independent predictors of stunting among schoolchildren. The early adolescent schoolchildren (AOR = 3.1, 95%CI: 1.86, 4.97, P < 0.001) and children from families with low (AOR = 2.1, 95%CI: 1.01, 4.15, P = 0.046) or moderate wealth (AOR = 2.3, 95%CI: 1.11, 4.77, P = 0.026) were significantly more wasted. Early adolescence (AOR = 1.8, 95%CI:1.14, 2.78, P = 0.011), female (AOR = 1.6, 95%CI: 1.03, 2.43, P = 0.038) and Al-Haimah Al-Dakheliah District (AOR = 3.4, 95%CI: 1.20, 9.55, P = 0.021) were independent risk factors for anemia. The study findings indicate highly focal prevalence of schistosomiasis in Sana'a Governorate with a public health significance that varies from low to high risk. Approximately half of schoolchildren were stunted, which was associated with S. mansoni infection and early adolescence. One quarter of schoolchildren were wasted with early adolescent schoolchildren and children from poor families being at high risk of wasting. Anemia was a moderate public health threat affecting the female and the early adolescent schoolchildren. The study suggests the implementation of control measures to combat schistosomiasis and integrated diseases control programmes to improve the health status of schoolchildren in Sana'a Governorate.

Onchocerca volvulus infection in Tihama region - west of Yemen: Continuing transmission in ivermectin-targeted endemic foci and unveiled endemicity in districts with previously unknown status.

BACKGROUND: Onchocerciasis in Yemen is one of the most neglected diseases, where baseline estimates of onchocerciasis and monitoring of the impact of ivermectin regularly administered to the affected individuals on its transmission are lacking. Therefore, this study aimed to determine the anti-Ov16 IgG4 seroprevalence among local communities of Hodeidah and Al-Mahwit governorates of Tihama region. The factors possibly associated with previous exposure to infection were also studied. METHODOLOGY/PRINCIPAL FINDINGS: This cross-sectional study was conducted in two ivermectin-targeted districts endemic for onchocerciasis in Hodeidah and Al-Mahwit and two untargeted districts with unknown previous endemicity in Hodeidah between February and July 2017. For 508 residents sampled by a multi-stage random approach, data were collected and blood specimens were screened for anti-Ov16 IgG4 using the SD BIOLINE Onchocerciasis IgG4 rapid tests. The study revealed an overall anti-Ov16 IgG4 rate of 18.5% (94/508) in all surveyed districts, with 10.2% (12/118) of children aged ≤10 years being seropositive. Moreover, rates of 8.0% (4/50) and 6.1% (4/66) were found in districts not officially listed as endemic for the disease. Multivariable analysis confirmed the age of more than ten years and residing within a large family as the independent predictors of exposure to infection. CONCLUSIONS/SIGNIFICANCE: Onchocerciasis transmission is still ongoing as supported by the higher anti-Ov16 IgG4 seroprevalence rate among children aged ≤10 years compared to that (<0.1%) previously set by the World Health Organization as a serologic criterion for transmission interruption. Further large-scale studies combining serologic and entomologic criteria are recommended for the mapping of O. volvulus in human and blackfly populations in endemic foci and their neighboring areas of uncertain endemicity. In addition, ivermectin distribution, coverage and impact on disease transmission need to be continually assessed.

Access to and use of long-lasting insecticidal nets and factors associated with non-use among communities in malaria-endemic areas of Al Hudaydah governorate in the Tihama region, west of Yemen.

BACKGROUND: Universal coverage of the targeted malaria-endemic areas with long-lasting insecticidal nets (LLINs) is implemented as one of the key interventions for malaria control and elimination in Yemen. In 2013, through a mass campaign, LLINs were distributed to the targeted communities in Al Hudaydah governorate. This study aimed to assess the ownership of, access to, and use of LLINs. It also aimed to identify factors associated with not using LLINs in malaria-endemic areas of Al Hudaydah in the Tihama region, west of Yemen. METHODS: A cross-sectional survey was conducted in four districts (Ad Durayhimi, Al Marawi'ah, Al Mansuriyah and Bayt Al Faqiah) in Al Hudaydah during February 2016. A total of 701 households were included in this study. Data on socio-demographic characteristics and availability of LLINs were collected by interview and observation. Four indicators for malaria prevention using LLINs; proportion of households with at least one LLIN, proportion of households with at least one LLIN for every two people, proportion of population with access to LLINs in the surveyed households and proportion of population who slept under LLINs the previous night of the survey were calculated as indicated by Roll Back Malaria Monitoring and Evaluation Reference Group. Use to access ratio was assessed. Factors associated with not using LLINs among people with access were also investigated. RESULTS: Of 701 households with 4900 de facto population, ownership of at least one LLIN was 90.6%, while 24.1% owned at least one for every two people during the survey in 2016. The overall proportion of people with access to LLINs was 51.5% (95% CI 50.1-52.9). Only 19.0% (95% CI 17.9-20.1) slept under LLINs the night before the survey and the overall use to access ratio was 0.37. The proportions of children under 5 years of age with access to and use of LLINs were 13.7 and 42.5%, respectively. On the other hand, the proportions of pregnant women with access to and use of LLINs were 16.4 and 20.0%, respectively. Multivariable analysis identified that people living in Al Mansuriyah district [adjusted odds ratio (AOR) = 3.29, 95% confidence interval (CI)  1.35-8.01; P = 0.009)], having three or more damaged LLINs in the house (AOR = 2.76, 95% CI 1.79-4.25; P < 0.001), aged between 16 and 45 years old (AOR = 2.17, 95% CI 1.26-3.75; P = 0.005) or older (AOR = 2.17, 95% CI 1.09-4.29; P = 0.026) and living in huts (AOR = 1.59, 95% CI 1.09-2.32; P = 0.015) were significantly less likely to use LLINs. CONCLUSIONS: This study shows a low LLIN access rate among local communities targeted for universal LLIN coverage in Al Hudaydah, a malaria-endemic area of high transmission. This finding necessitates additional distribution channels following mass campaigns to maintain the universal coverage. Reduced use of LLINs among people with access in these communities together with the identified risks of non-use highlight the importance of conducting behaviour change communication campaigns to enhance using LLINs in areas with universal coverage.

A community-based survey of Toxoplasma gondii infection among pregnant women in rural areas of Taiz governorate, Yemen: the risk of waterborne transmission.

BACKGROUND: Toxoplasma gondii is a zoonotic coccidian parasite causing morbidity and mortality. In Yemen, T. gondii infection has been reported among pregnant women seeking healthcare in the main cities. However, no data are available on the prevalence of T. gondii infection and its associated risk factors among pregnant women in the rural communities of the country. Thus, the present study aimed to determine the seroprevalence of T. gondii and identify its risk factors among pregnant women in the rural communities of Taiz governorate, Yemen. METHODS: A total of 359 pregnant women living in the rural communities of Taiz governorate were enrolled in this study by house-to-house visits. Data were collected using a pre-designed questionnaire, and blood samples were collected and tested for the detection of anti- T. gondii IgM and IgG antibodies by enzyme-linked immunosorbent assay. RESULTS: The prevalence of T. gondii infection among pregnant women in this study was 46.2% (166/359). Bivariate analysis identified the age of ≥ 30 years (odds ratio [OR] = 1.7; 95% confidence interval [CI] = 1.09-2.65, P = 0.019) and unimproved water sources (OR = 2.2; 95% CI = 1.10-4.55, P = 0.023) as factors associated with T. gondii infection among pregnant women. The multivariable analysis, however, identified unimproved water sources as an independent risk factor (adjusted OR = 2.4; 95% CI = 1.16-5.0, P = 0.018) associated with T. gondii infection among pregnant women. CONCLUSIONS: Pregnant women in the rural communities of Taiz, Yemen are at high risk of contracting T. gondii infection. Unimproved water sources (wells, water streams and water tanks) are significantly associated with T. gondii infection and should be considered in prevention and control strategies, especially among pregnant women.

A household-based survey of knowledge, attitudes and practices towards dengue fever among local urban communities in Taiz Governorate, Yemen.

BACKGROUND: Yemen has witnessed several dengue fever outbreaks coincident with the social unrest and war in the country. The aim of the present study was to describe the knowledge, attitudes and practices (KAPs) of at-risk urban populations residing in Taiz, southwest of Yemen. In addition, factors possibly associated with poor preventive practices were investigated. METHODS: A household-based, cross-sectional survey was conducted in three urban districts encompassing 383 households. Data on the socio-demographic characteristics and KAPs of the participating household heads were collected using a pre-designed, structured questionnaire. The association of socio-demographic characteristics, knowledge and attitudes of the population with poor preventive practices against dengue fever was then analyzed using logistic regression. RESULTS: More than 90.0 % of respondent household heads had correct knowledge about fever, headache and joint pain as common signs and symptoms of dengue fever. Moreover, muscular pain and bleeding were perceived by more than 80.0 % of the respondents as being associated with dengue fever; however, only 65.0 % of the respondents reported skin rash as a sign of dengue fever. More than 95.0 % of respondents agreed about the seriousness and possible transmission of dengue fever; however, negative attitudes regarding the facts of being at risk of the disease and that the infection is preventable were expressed by 15.0 % of respondents. Despite the good level of knowledge and attitudes of the respondent population, poor preventive practices were common. Bivariate analysis identified poor knowledge of dengue signs and symptoms (OR = 2.1, 95 % CI = 1.24-3.68; P = 0.005) and its vector (OR = 2.1, 95 % CI = 1.14-3.84; P = 0.016) as factors significantly associated with poor preventive practices. However, multivariable analysis showed that poor knowledge of the vector is an independent predictor of poor preventive practices of the population (adjusted OR = 2.1, 95 % CI = 1.14-3.84; P = 0.018). CONCLUSION: The majority of people in urban communities of Taiz have a clear understanding of most signs/symptoms of dengue fever as well as positive attitudes towards the seriousness and possible transmissibility of dengue fever. However, negative attitudes regarding their perception of the risk and possible prevention of the infection are prevailing among a small proportion of the population and need to be targeted by educational campaigns. It appears that the good level of the population knowledge of the signs/symptoms of dengue fever and the factors contributing to the spread and control of its vectors did not translate into good practices.

School-based prevalence of intestinal parasitic infections and associated risk factors in rural communities of Sana'a, Yemen.

Yemen is a developing country overwhelmed with a triad of poverty, diseases and social conflicts. Moreover, the majority of its population live in rural communities and suffer from intestinal parasitic infections (IPIs). Therefore, the present school-based, cross-sectional survey aimed to detect the prevalence of such infections and associated risk factors among schoolchildren in the rural communities of Bani Alharith, Hamdan and Bani Hushaysh districts of Sana'a, north of Yemen. Socio-demographic data and certain behavioral risk factors as well as stool samples were collected from 1218 schoolchildren from ten randomly schools in the study area. Fresh stool samples were examined for parasites by direct saline and iodine preparations and after concentration with formol-ether technique. The overall prevalence of IPIs was 54.8%, with a higher frequency of protozoal than helminthic infections (37.6 vs. 17.2%, respectively). Parasite species recovered were Entameba histolytica (21.5%), Giardia lamblia (16.1%), Ascaris lumbricoides (8.3%), Hymenolepis nana (5.3%), Schistosoma mansoni (2.6%), Trichuris trichiura (0.5%) and Enterobius vermicularis (0.4%). Univariate analysis showed that the male gender and illiteracy of fathers and/or mothers were the socio-demographic factors significantly associated with higher infection rates. The illiteracy of mothers was also confirmed as an independent risk factor by multivariable analysis. On the other hand, not washing hands before eating, not washing fruits and vegetables before consumption, eating uncovered food and not clipping fingernails were the risk behaviors significantly associated with higher infection rates, with the last three ones being confirmed as independent risk factors. Therefore, control measures should include regular treatment of protozoal infections and deworming of schoolchildren, promotion of hygiene in rural schools through health education programs, regular inspection of schoolchildren for personal hygiene practices and the provision of a healthy school infrastructure.

Toxoplasma gondii infection among pregnant women in Yemen: Factors associated with high seroprevalence.

INTRODUCTION: Although toxoplasmosis is an important public health problem, there is scarcity of data on the disease available from Yemen. A cross-sectional survey was conducted in health facilities to determine seroprevalence of Toxoplasma gondii and associated risk factors among pregnant women in Sana'a, the capital city of Yemen. METHODOLOGY: A total of 593 pregnant women were included and examined for anti-T. gondii antibodies (Ab) using enzyme-linked immunosorbent assay. Bio and socio-demographic data were collected by pre-tested structured questionnaires through face-to-face interviews. RESULTS: The overall seroprevalence of T. gondii was 45.4% (95% confidence interval: 41%-49%). The prevalence of anti-T. gondii IgG and IgM was 43.7 (95% CI: 40-%48%) and 9.1% (95% CI: 7%-12%), respectively. About 7.4 (95% CI: 6%-10%) of pregnant women were seropositive for both IgG and IgM Abs. Multivariate logistic regression analysis identified the following risk factors for toxplasmosis (IgG and/or IgM): age ≥ 25 years (adjusted OR: 2.02, 95% CI: 1.44-2.84, p < 0.001), rearing cats in the house (OR: 1.75, 95% CI: 1.20-2.55, p = 0.004), and contact with soil (OR: 1.90, 95% CI: 1.32-2.75, p = 0.001). CONCLUSIONS: The study reported high seroprevalence among pregnant women in Sana'a, Yemen, with a high proportion of pregnant women having a possibility of acute toxoplasmosis. This highlights the need for including routine screening for T. gondii in pregnant women in the country's antenatal clinics. In addition, health education on the mode of transmission of toxoplasmosis should be provided for pregnant women in Yemen.

First Molecular Characterization of Leishmania Species Causing Visceral Leishmaniasis among Children in Yemen.

Visceral leishmaniasis (VL) is a debilitating, often fatal disease caused by Leishmania donovani complex; however, it is a neglected tropical disease. L. donovani complex comprises two closely related species, L. donovani that is mostly anthroponotic and L. infantum that is zoonotic. Differentiation between these two species is critical due to the differences in their epidemiology and pathology. However, they cannot be differentiated morphologically, and their speciation using isoenzyme-based methods poses a difficult task and may be unreliable. Molecular characterization is now the most reliable method to differentiate between them and to determine their phylogenetic relationships. The present study aims to characterize Leishmania species isolated from bone marrows of Yemeni pediatric patients using sequence analysis of the ribosomal internal transcribed spacer-1 (ITS1) gene. Out of 41 isolates from Giemsa-stained bone marrow smears, 25 isolates were successfully amplified by nested polymerase chain reaction and sequenced in both directions. Phylogenetic analysis using neighbor joining method placed all study isolates in one cluster with L. donovani complex (99% bootstrap). The analysis of ITS1 for microsatellite repeat numbers identified L. infantum in 11 isolates and L. donovani in 14 isolates. These data suggest the possibility of both anthroponotic and zoonotic transmission of VL-causing Leishmania species in Yemen. Exploring the possible animal reservoir hosts is therefore needed for effective control to be achieved.

The prospect of malaria elimination in the Arabian Peninsula: a population genetic approach.

BACKGROUND: In the Arabian Peninsula malaria control is progressing steadily, backed by adequate logistic and political support. As a result, transmission has been interrupted throughout the region, with exception of limited sites in Yemen and Saudi Arabia. Here we examined Plasmodium falciparum parasites in these sites to assess if the above success has limited diversity and gene flow. METHODS: We examined 108 P. falciparum isolates in three sites in Yemen (Taiz, Dhamar and Hodeidah) and 91 isolates from Saudi Arabia (Jazan). Nine microsatellites were analyzed for allelic diversity, multi-locus haplotype and inter-population differentiation. RESULTS: Diversity at each locus (unbiased heterozygosity [H]) was relatively lower in Yemen; (Hodeidah, H=0.615, Taiz, H=0.66, Dhamar, H=0.481), compared to Saudi Arabia (Jazan, H=0.76). Microsatellites were distributed widely and private alleles, detected in a single population, were rare. Pairwise comparisons revealed that parasites population in Dhamar was relatively distanced (FST=0.19). However, Taiz (Yemen) (FST=0.065) and Hodeidah (FST=0.107) populations were closer to that in Jazan (Saudi Arabia). Nonetheless, parasites in the four sites can be considered as one population. CONCLUSION: Although malaria risk in Saudi Arabia has been cut considerably, the extent of diversity and parasite genetic structure are indicative of a large population size. Elimination strategy should target demographic factors that favor parasite dispersal and flow of imported malaria.

Subtype distribution of Blastocystis isolates in Sebha, Libya.

BACKGROUND: Blastocystis is a genetically diverse and a common intestinal parasite of humans with a controversial pathogenic potential. This study was carried out to identify the Blastocystis subtypes and their association with demographic and socioeconomic factors among outpatients living in Sebha city, Libya. METHODS/FINDINGS: Blastocystis in stool samples were cultured followed by isolation, PCR amplification of a partial SSU rDNA gene, cloning, and sequencing. The DNA sequences of isolated clones showed 98.3% to 100% identity with the reference Blastocystis isolates from the Genbank. Multiple sequence alignment showed polymorphism from one to seven base substitution and/or insertion/deletion in several groups of non-identical nucleotides clones. Phylogenetic analysis revealed three assemblage subtypes (ST) with ST1 as the most prevalent (51.1%) followed by ST2 (24.4%), ST3 (17.8%) and mixed infections of two concurrent subtypes (6.7%). BLASTOCYSTIS: ST1 infection was significantly associated with female (P = 0.009) and low educational level (P = 0.034). ST2 was also significantly associated with low educational level (P= 0.008) and ST3 with diarrhoea (P = 0.008). CONCLUSION: Phylogenetic analysis of Libyan Blastocystis isolates identified three different subtypes; with ST1 being the predominant subtype and its infection was significantly associated with female gender and low educational level. More extensive studies are needed in order to relate each Blastocystis subtype with clinical symptoms and potential transmission sources in this community.

Mutant Plasmodium falciparum chloroquine resistance transporter in Hodeidah, Yemen: association with parasitologic indices and treatment-seeking behaviors.

Malaria still represents a major health problem in Yemen, particularly in Hodeidah, despite continuing efforts to eliminate it. With the absence of clinically proven vaccines, chemotherapy with antimalarials is still greatly needed. Chloroquine (CQ) has been popular as the drug of choice for malaria control. However, Plasmodium falciparum resistance to CQ has been one of the main obstacles in malaria control and elimination. Although CQ is no longer the recommended antimalarial chemotherapy, it has remained the number one over-the-counter antimalarial drug in many endemic areas, including Yemen, and is still used for self-medication. In addition, promising reports on CQ efficacy reversal in many African countries brought it again into the scene. This has led to a growing interest in the possibility of its re-introduction, particularly with the concerns raised about the parasite resistance to artemisinin-based combination therapies. Therefore, the present study aimed at analyzing the CQ-associated pfcrt 76T mutation in P. falciparum isolates from patients with uncomplicated falciparum malaria in Hodeidah, west of Yemen. The association of treatment-seeking behaviors and antimalarial drug use with the pfcrt 76T mutant allele was also studied. It was revealed that there is still a sustained high frequency of this molecular marker among parasite isolates associated with younger age, decreased parasite density and the presence of gametocytes in blood. Delay in seeking treatment and frequent use of antimalarials were the behaviors significantly associated with the presence of the pfcrt 76T mutant allele among patients reporting a history of malaria treatment.

Genetic diversity of Plasmodium falciparum and distribution of drug resistance haplotypes in Yemen.

BACKGROUND: Despite evident success of malaria control in many sites in the Arabian Peninsula, malaria remains endemic in a few spots, in Yemen and south-west of Saudi Arabia. In addition to local transmission, imported malaria sustains an extra source of parasites that can challenge the strengths of local control strategies. This study examined the genetic diversity of Plasmodium falciparum in Yemen and mutations of drug resistant genes, to elucidate parasite structure and distribution of drug resistance genotypes in the region. METHODS: Five polymorphic loci (MSP-2, Pfg377 and three microsatellites on chromosome 8) not involved in anti-malarial drug resistance, and four drug resistant genes (pfcrt, pfmdr1, dhfr and dhps) were genotyped in 108 P. falciparum isolates collected in three sites in Yemen: Dhamar, Hodeidah and Taiz. RESULTS: High diversity was seen in non-drug genes, pfg377 (He = 0.66), msp-2 (He = 0.80) and three microsatellites on chr 8, 7.7 kb (He = 0.88), 4.3 kb (He = 0.77) and 0.8 kb (He = 0.71). There was a high level of mixed-genotype infections (57%), with an average 1.8 genotypes per patient. No linkage disequilibrium was seen between drug resistant genes and the non-drug markers (p < 0.05). Genetic differentiation between populations was low (most pair-wise FST values <0.03), indicating extensive gene flow between the parasites in the three sites. CONCLUSION: The high diversity of P. falciparum in Yemen is indicative of a large parasite reservoir, which represents a challenge to control efforts. The presence of two distinct pfcrt genotype, CVIET and SVMNT, suggests that chloroquine resistance can possibly be related to a migratory path from Africa and Asia. The absence of the triple mutant dhfr genotype (IRN) and dhps mutations supports the use of artesunate + sulphadoxine-pyrimethamine as first-line therapy. However, the prevalent pfmdr1 genotype NFSND [21%] has previously been associated with tolerance/resistance response to artemisinin combination therapy (ACT). Regular surveys are, therefore, important to monitor spread of pfmdr1 and dhfr mutations and response to ACT.

The suitability of P. falciparum merozoite surface proteins 1 and 2 as genetic markers for in vivo drug trials in Yemen.

BACKGROUND: The accuracy of the conclusions from in vivo efficacy anti-malarial drug trials depends on distinguishing between recrudescences and re-infections which is accomplished by genotyping genes coding P. falciparum merozoite surface 1 (MSP1) and MSP2. However, the reliability of the PCR analysis depends on the genetic markers' allelic diversity and variant frequency. In this study the genetic diversity of the genes coding for MSP1 and MSP2 was obtained for P. falciparum parasites circulating in Yemen. METHODS: Blood samples were collected from 511 patients with fever and screened for malaria parasites using Giemsa-stained blood films. A total 74 samples were infected with P. falciparum, and the genetic diversity was assessed by nested PCR targeting Pfmsp1 (Block2) and Pfmsp2 (block 3). RESULTS: Overall, 58%, 28% and 54% of the isolates harboured parasites of the Pfmsp1 K1, MAD20 and RO33 allelic families, and 55% and 89% harboured those of the Pfmsp2 FC27 and 3D7 allelic families, respectively. For both genetic makers, the multiplicity of the infection (MOI) was significantly higher in the isolates from the foothills/coastland areas as compared to those from the highland (P<0.05). Pfmsp2 had higher number of distinct allelic variants than Pfmsp1 (20 vs 11). The expected heterozygosity (HE) for Pfmsp1 and Pfmsp2 were 0.82 and 0.94, respectively. Nonetheless, a bias in the frequency distribution of the Pfmsp1 allelic variants was noted from all areas, and of those of Pfmsp2 in the samples collected from the highland areas. CONCLUSIONS: Significant differences in the complexity and allelic diversity of Pfmsp1 and Pfmsp2 genes between areas probably reflect differences in the intensity of malaria transmission. The biased distribution of allelic variants suggests that in Yemen Pfmsp1 should not be used for PCR correction of in vivo clinical trials outcomes, and that caution should be exercised when employing Pfmsp2.

Cases of hydatidosis in patients referred to Governmental hospitals for cyst removal in Sana'a City, Republic of Yemen.

Hydatidosis is a parasitic infestation caused by Echinococcus granulosus. This disease is endemic in many countries including Yemen. The present review article aims to have a glimpse at the present status of hydatidosis in Yemen. This is the first descriptive study, investigating recorded cases of hydatidosis from the five main governmental hospitals in the capital Sana'a city, over a longer period starting from 2001 and ending in 2008. A total of 796 medical records of patients referred to the five main governmental hospitals in Sana'a city for cyst removal, were studied. Of these cases 482 were females and 314 were males. Their mean age was 30.0 ± 16.9 years. Information regarding the location of the cyst in the body, age, sex and residence of each patient was recorded. A higher infection rate was found in females than males (60.6% and 39.4%, respectively). Single organ involvement was observed in 98.6% cases, among which, the most frequent localizations were the liver (60.8%) followed by the lung (24.7%). Cases of hydatidosis appeared to increase during the period 2001-2008, with the lowest number (n=26) and the highest number (n=140) recorded in 2001 and 2007, respectively. We conclude that the risk of hydatidosis is still high in Yemen, where street or stray dogs move freely down town and the population should be aware about the role of dogs in the transmission of this disease. Hospital records provide a useful indication of infection expressed as annual rate of hospital cases. Finally, the collaboration of Public Health Authorities, the Veterinary Medical Authorities and the Environmental Affairs Authorities is a must to control this disease.

Loop-mediated isothermal amplification (LAMP) for malarial parasites of humans: would it come to clinical reality as a point-of-care test?

Loop-mediated isothermal amplification (LAMP) is a novel molecular method that accelerates and facilitates DNA amplification and detection under isothermal conditions. It represents a revolution in molecular biology by reducing the high cost, turnaround time and technicality of polymerase chain reaction and other amplification methods. It has been applied for the diagnosis of a variety of viral, bacterial, parasitic and other diseases in the biomedical field. LAMP has been involved in studies concerning the diagnosis of malaria which is still a major cause of morbidity and mortality in different parts of the world. For the success attained with this technology to diagnose human malaria, is it time to think that LAMP-based point-of-care diagnostics come to application to support the diagnosis of clinical malaria cases? The present review deals with the use of LAMP in the diagnosis of malaria and related investigations to make a view on what has been investigated and highlights the future perspectives regarding the possible applications of LAMP in diagnosis of the disease.

Microbial control of malaria: biological warfare against the parasite and its vector.

Microbial applications in malaria transmission control have drawn global attention. Mosquito midgut microbiota can modulate vector immunity and block Plasmodium development. Paratransgenic manipulation of bacterial symbionts and Wolbachia can affect reproductive characteristics of mosquitoes. Bacillus-based biolarvicides can control mosquito larvae in different breeding habitats, but their effectiveness differs according to the type of formulation applied, and the physical and ecological conditions of the environment. Entomopathogenic fungi show promise as effective and evolution-proof agents against adult mosquitoes. In addition, transgenic fungi can express anti-plasmodial effector molecules that can target the parasite inside its vector. Despite showing effectiveness in domestic environments as well as against insecticide-resistant mosquitoes, claims towards their deployability in the field and their possible use in integrated vector management programmes have yet to be investigated. Viral pathogens show efficacy in the interruption of sporogonic development of the parasite, and protozoal pathogens exert direct pathogenic potential on larvae and adults with substantial effects on mosquito longevity and fecundity. However, the technology required for their isolation and maintenance impedes their field application. Many agents show promising findings; however, the question remains about the epidemiologic reality of these approaches because even those that have been tried under field conditions still have certain limitations. This review addresses aspects of the microbial control of malaria between proof-of-concept and epidemiologic reality.