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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.

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.

Background and aims: Toxoplasmosis is caused as a result of intracellular protozoan organism, Toxoplasma gondii, and is a widespread disease. Toxoplasmosis can progress to a serious systemic disease (congenital form); once a mother becomes infected for the first time for the period of pregnancy, a transient haematopoietic parasite can appear with focal lesions produced within the placenta, thus infecting the fetus. Therefore, this cross-sectional study aimed at assessing the immune status of the pregnant woman towards toxoplasmosis, determining the prevalence of T. gondii in Amran city -Yemen, and identifying the potential risk factors that affect the pregnant woman towards toxoplasmosis. Material and methods: This study included 280 pregnant women who were admitted to maternity clinics at Amran General Hospital and Family Health Center in Amran City -Yemen during the period from December 2016 to June 2017. The questionnaire was used to obtain relevant demographic data and potential risk factors. Then, blood samples were collected from pregnant women and tested for identification of IgM and IgG antibodies against T. gondii using the ELISA technique. Results: The total positive rate for IgM antibodies was 3.6%, while for IgG was 27.9%. A high IgM rate equal to 5.3% was observed in the 31-35 age group, and a higher IgG rate (36.8%) was observed in the same age group. There were 69.3% of pregnant women susceptible to infection with toxoplasmosis, 2.9% recently infected, 27.1% protected, and 0.7% had secondary infection. The high rate of IgM positivity was observed in the second trimester of pregnancy (4.7%), while the lowest rate was in the third trimester of pregnancy (2%). A slight increase in the rate of IgG positivity was observed in the third trimester of pregnancy (32%). There was a significant association between contact with animals such as cats and infection with toxoplasmosis, where OR =5.4, CI =1.01-30.13, χ2 =4.59, p =0.04. While there was no significant association with the other studied factors. Conclusion: Yemen has less seropositivity than other Arab and African countries as a result of variation in risk factors. Data on T. gondii infection during pregnancy is scarce in numerous countries, in particular where there is a lack of political constancy such as Yemen. The risk factors identified included proximity to the cats and domistic animals, and increased maternal age. Toxoplasmosis in pregnancy in Yemen may be a really underestimated health problem. More research is needed. Peer Review History: Received 19 March 2021; Revised 12 April; Accepted 28 April, Available online 15 May 2021 UJPR follows the most transparent and toughest ‘Advanced OPEN peer review’ system. The identity of the authors and, reviewers will be known to each other. This transparent process will help to eradicate any possible malicious/purposeful interference by any person (publishing staff, reviewer, editor, author, etc) during peer review. As a result of this unique system, all reviewers will get their due recognition and respect, once their names are published in the papers. We expect that, by publishing peer review reports with published papers, will be helpful to many authors for drafting their article according to the specifications. Auhors will remove any error of their article and they will improve their article(s) according to the previous reports displayed with published article(s). The main purpose of it is ‘to improve the quality of a candidate manuscript’. Our reviewers check the ‘strength and weakness of a manuscript honestly’. There will increase in the perfection, and transparency. Received file: Reviewer's Comments: Average Peer review marks at initial stage: 6.5/10 Average Peer review marks at publication stage: 7.5/10 Reviewer(s) detail: Rola Jadallah, Arab American University, Palestine, rola@aauj.edu Prof. Dr. Gorkem Dulger, Duzce University, Turkey, gorkemdulger@yandex.com Similar Articles: PREVALENCE AND RISK FACTORS ASSOCIATED WITH VULVOVAGINAL CANDIDIASIS DURING PREGNANCY IN SANA'A, YEMEN VULVOVAGINAL CANDIDIASIS PREVALENCE AMONG PREGNANT WOMEN IN DIFFERENT HOSPITALS IN IBB, YEMEN

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., Author summary 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. Information was collected using a standard questionnaire. S. mansoni was identified by microscopic examination of Kato-Katz fecal smear. Hemoglobin concentration and anthropometric measurements were estimated using standard methods. The prevalence of S. mansoni was highly focal (1.4–33.9%). Schoolchildren living in households without tap water had significantly high infection rate of S. mansoni. The prevalence of wasting, stunting, underweight and anemia among schoolchildren was 25.0%, 45.8%, 27.3% and 31.7%, respectively. Although S. mansoni was significantly associated with stunting, no significant association was found between S. mansoni and wasting, underweight or anemia. The stunting was higher among early adolescent than young schoolchildren. The early adolescent schoolchildren and children from families with low or moderate wealth were significantly more wasted than schoolchildren from families with high wealth. Schoolchildren in the early adolescence, being female or living in Al-Haimah Al-Dakheliah District were at high risk of anemia. The study suggests implementation of control measures to combat schistosomiasis and integrated diseases control programmes to improve the health status of schoolchildren in Sana’a Governorate.

Objective: To determine the prevalence and risk factors for Trichomonas vaginalis infection among pregnant women seeking health care in Sana’a city. Methods: This cross-sectional study was conducted among pregnant women seeking health care in private clinics and governmental primary health care centers in Sana’a during the period from October 2014 to June 2015. Data on sociodemographic and clinical characteristics as well as possible risk factors of T. vaginalis infection were collected from 287 pregnant women using a pre-designed questionnaire by trained researchers. Vaginal discharges were then collected and examined microscopically for motile T. vaginalis trophozoites. Results: The overall prevalence of T. vaginalis infection was 11.1%, with a higher infection rate among pregnant women aged 26-40 years old, housewives and rural residents. Age, literacy status and the age at first sexual intercourse were significantly associated with T. vaginalis infection among pregnant women. Presence of vaginal discharge (OR = 8.33; 95% CI: 2.47–28.03, P

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.

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

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. Electronic supplementary material The online version of this article (doi:10.1186/s40249-017-0243-0) contains supplementary material, which is available to authorized users.

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.

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.

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.

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 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.

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.

BackgroundDespite 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 ofPlasmodium falciparumin Yemen and mutations of drug resistant genes, to elucidate parasite structure and distribution of drug resistance genotypes in the region.MethodsFive polymorphic loci (MSP-2,Pfg377and three microsatellites on chromosome 8) not involved in anti-malarial drug resistance, and four drug resistant genes (pfcrt,pfmdr1,dhfranddhps) were genotyped in 108P.falciparumisolates collected in three sites in Yemen: Dhamar, Hodeidah and Taiz.ResultsHigh 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 FSTvalues There was a high prevalence of mutations inpfmdr1,pfcrtanddhfr; with four mutantpfmdr1genotypes (NFCDD[57%], NFSND[21%], YFCDD[13%] and YFSND[8% ]), two mutantpfcrtgenotypes (CVIET[89%] and SVMNT[4%]) and one mutantdhfrgenotype (ICNI[53.7%]). However, nodhpsmutations were detected.ConclusionThe high diversity ofP.falciparumin Yemen is indicative of a large parasite reservoir, which represents a challenge to control efforts. The presence of two distinctpfcrtgenotype, CVIET and SVMNT, suggests that chloroquine resistance can possibly be related to a migratory path from Africa and Asia. The absence of the triple mutantdhfrgenotype (IRN) anddhpsmutations supports the use of artesunate + sulphadoxine-pyrimethamine as first-line therapy. However, the prevalentpfmdr1genotype 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.

International audience; 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

BackgroundBlastocystis 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/findingsBlastocystis 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%).BlastocystisST1 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).ConclusionPhylogenetic 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.

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.

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.

A community-based cross-sectional study was carried out among Aboriginal schoolchildren aged 7–12 years living in remote areas in Pos Betau, Pahang, Malaysia to investigate the potential determinants influencing the cognitive function and educational achievement of these children. Cognitive function was measured by intelligence quotient (IQ), while examination scores of selected school subjects were used in assessing educational achievement. Blood samples were collected to assess serum Fe status. All children were screened for soil-transmitted helminthes. Demographic and socio-economic data were collected using pre-tested questionnaires. Almost two-thirds (67·6 %) of the subjects had poor IQ and most of them (72·6 %) had insufficient educational achievement. Output of the stepwise multiple regression model showed that poor IQ was significantly associated with low household income which contributed the most to the regression variance (r2 0·059; P = 0·020). Low maternal education was also identified as a significant predictor of low IQ scores (r2 0·042; P = 0·043). With educational achievement, Fe-deficiency anaemia (IDA) was the only variable to show significant association (r2 0·025; P = 0·015). In conclusion, the cognitive function and educational achievement of Aboriginal schoolchildren are poor and influenced by household income, maternal education and IDA. Thus, effective and integrated measures to improve the nutritional and socio-economic status of rural children would have a pronounced positive effect on their education.

title/Between June 2008 and March 2009, a cross-sectional study of human malaria was carried out in four governorates of Yemen, two (Taiz and Hodiedah) representing the country's highlands and the others (Dhamar and Raymah) the country's coastal plains/foothills. The main aims were to determine the prevalences of Plasmodium infection among 455 febrile patients presenting for care at participating health facilities and to investigate the potential risk factors for such infection. Malarial infection was detected in 78 (17·1%) of the investigated patients and was more likely to be detected among the febrile patients from the highlands than among those presenting in the coastal plains/foothills (22·6% v.13·9%; χ(2)=10·102; P=0·018). Binary logistic-regression models identified low household income [odds ratio (OR)=13·52; 95% confidence interval (CI)=2·62-69·67; P=0·002], living in a household with access to a water pump (OR=4·18; CI=1·60-10·96; P=0·004) and living in a household near a stream (OR=4·43; CI=1·35-14·56; P=0·014) as significant risk factors for malarial infection in the highlands. Low household income was the only significant risk factor identified for such infection in the coastal plains and foothills (OR = 8·20; CI=1·80-37·45; P=0·007). It is unclear why febrile patients in the highlands of Yemen are much more likely to be found to have malarial infection than their counterparts from the coastal plains and foothills. Although it is possible that malarial transmission is relatively intense in the highlands, it seems more likely that, compared with those who live at lower altitudes, those who live in the highlands are less immune to malaria, and therefore more likely to develop febrile illness following malarial infection. Whatever the cause of the symptomatic malarial infection commonly found in the highlands of Yemen, it is a matter of serious concern that should be addressed in the national strategy to control malaria.

Background The emergence of Plasmodium knowlesi in humans, which is in many cases misdiagnosed by microscopy as Plasmodium malariae due to the morphological similarity has contributed to the needs of detection and differentiation of malaria parasites. At present, nested PCR targeted on Plasmodium ssrRNA genes has been described as the most sensitive and specific method for Plasmodium detection. However, this method is costly and requires trained personnel for its implementation. Loop-mediated isothermal amplification (LAMP), a novel nucleic acid amplification method was developed for the clinical detection of P. knowlesi. The sensitivity and specificity of LAMP was evaluated in comparison to the results obtained via microscopic examination and nested PCR. Methods LAMP assay was developed based on P. knowlesi genetic material targeting the apical membrane antigen-1 (AMA-1) gene. The method uses six primers that recognize eight regions of the target DNA and it amplifies DNA within an hour under isothermal conditions (65°C) in a water-bath. Results LAMP is highly sensitive with the detection limit as low as ten copies for AMA-1. LAMP detected malaria parasites in all confirm cases (n = 13) of P. knowlesi infection (sensitivity, 100%) and none of the negative samples (specificity, 100%) within an hour. LAMP demonstrated higher sensitivity compared to nested PCR by successfully detecting a sample with very low parasitaemia (< 0.01%). Conclusion With continuous efforts in the optimization of this assay, LAMP may provide a simple and reliable test for detecting P. knowlesi malaria parasites in areas where malaria is prevalent.

This study was conducted to evaluate the modification of the usual Gram-chromotrope staining technique developed in-house known as Gram-chromotrope Kinyoun (GCK) in comparison with the Weber Modified Trichrome (WMT) staining technique; as the reference technique. Two hundred and ninety fecal specimens received by the Microbiology Diagnostic Laboratory of Hospital Universiti Kebangsaan Malaysia were examined for the presence of microsporidial spores. The sensitivity and specificity of GCK compared to the reference technique were 98% and 98.3%, respectively. The positive and negative predictive values were 92.5% and 99.6%, respectively. The agreement between the reference technique and the GCK staining technique was statistically significant by Kappa statistics (K = 0.941, P < 0.001). It is concluded that the GCK staining technique has high sensitivity and specificity in the detection of microsporidial spores in fecal specimens. Hence, it is recommended to be used in the diagnosis of intestinal microsporidiosis.

Background Malaria is an endemic disease in Yemen and is responsible for 4.9 deaths per 100,000 population per year and 43,000 disability adjusted life years lost. Although malaria in Yemen is caused mainly by Plasmodium falciparum and Plasmodium vivax, there are no sequence data available on the two species. This study was conducted to investigate the distribution of the Plasmodium species based on the molecular detection and to study the molecular phylogeny of these parasites. Methods Blood samples from 511 febrile patients were collected and a partial region of the 18 s ribosomal RNA (18 s rRNA) gene was amplified using nested PCR. From the 86 positive blood samples, 13 Plasmodium falciparum and 4 Plasmodium vivax were selected and underwent cloning and, subsequently, sequencing and the sequences were subjected to phylogenetic analysis using the neighbor-joining and maximum parsimony methods. Results Malaria was detected by PCR in 86 samples (16.8%). The majority of the single infections were caused by P. falciparum (80.3%), followed by P. vivax (5.8%). Mixed infection rates of P. falciparum + P. vivax and P. falciparum + P. malariae were 11.6% and 2.3%, respectively. All P. falciparum isolates were grouped with the strain 3D7, while P. vivax isolates were grouped with the strain Salvador1. Phylogenetic trees based on 18 s rRNA placed the P. falciparum isolates into three sub-clusters and P. vivax into one cluster. Sequence alignment analysis showed 5-14.8% SNP in the partial sequences of the 18 s rRNA of P. falciparum. Conclusions Although P. falciparum is predominant, P. vivax, P. malariae and mixed infections are more prevalent than has been revealed by microscopy. This overlooked distribution should be considered by malaria control strategy makers. The genetic polymorphisms warrant further investigation.

Background Malaria remains a significant health problem in Yemen with Plasmodium falciparum being the predominant species which is responsible for 90% of the malaria cases. Despite serious concerns regarding increasing drug resistance, chloroquine is still used for the prevention and treatment of malaria in Yemen. This study was carried out to determine the prevalence of choloroquine resistance (CQR) of P. falciparum isolated from Yemen based on the pfcrt T76 mutation. Methods A cross-sectional study was carried out among 511 participants from four governorates in Yemen. Blood samples were screened using microscopic and species-specific nested PCR based on the 18S rRNA gene to detect and identify Plasmodium species. Blood samples positive for P. falciparum were used for detecting the pfcrt T76 mutation using nested-PCR. Results The prevalence of pfcrt T76 mutation was 81.5% (66 of 81 isolates). Coastal areas/foothills had higher prevalence of pfcrt T76 mutation compared to highland areas (90.5% vs 71.8%) (p = 0.031). The pfcrt T76 mutation had a significant association with parasitaemia (p = 0.045). Univariate analysis shows a significant association of pfcrt T76 mutation with people aged > 10 years (OR = 9, 95% CI = 2.3 - 36.2, p = 0.001), low household income (OR = 5, 95% CI = 1.3 - 19.5, p = 0.027), no insecticide spray (OR = 3.7, 95% CI = 1.16 - 11.86, p = 0.025) and not sleeping under insecticide treated nets (ITNs) (OR = 4.8, 95% CI = 1.38 - 16.78, p = 0.01). Logistic regression model confirmed age > 10 years and low household income as predictors of pfcrt T76 mutation in Yemen P. falciparum isolates. Conclusions The high prevalence of pfcrt T76 mutation in Yemen could be a predictive marker for the prevalence of P. falciparum CQR. This finding shows the necessity for an in-vivo therapeutic efficacy test for CQ. P. falciparum CQR should be addressed in the national strategy to control malaria.