The Sri Lankan paradox: high genetic diversity in Plasmodium vivax populations despite decreasing levels of malaria transmission.

Here we examined whether the recent dramatic decline in malaria transmission in Sri Lanka led to a major bottleneck in the local Plasmodium vivax population, with a substantial decrease in the effective population size. To this end, we typed 14 highly polymorphic microsatellite markers in 185 P. vivax patient isolates collected from 13 districts in Sri Lanka over a period of 5 years (2003-2007). Overall, we found a high degree of polymorphism, with 184 unique haplotypes (12-46 alleles per locus) and average genetic diversity (expected heterozygosity) of 0·8744. Almost 69% (n = 127) isolates had multiple-clone infections (MCI). Significant spatial and temporal differentiation (F ST = 0·04-0·25; P⩽0·0009) between populations was observed. The effective population size was relatively high but showed a decline from 2003-4 to 2006-7 periods (estimated as 45 661 to 22 896 or 10 513 to 7057, depending on the underlying model used). We used three approaches - namely, mode-shift in allele frequency distribution, detection of heterozygote excess and the M-ratio statistics - to test for evidence of a recent population bottleneck but only the low values of M-ratio statistics (ranging between 0·15-0·33, mean 0·26) were suggestive of such a bottleneck. The persistence of high genetic diversity and high proportion of MCI, with little change in effective population size, despite the collapse in demographic population size of P. vivax in Sri Lanka indicates the importance of maintaining stringent control and surveillance measures to prevent resurgence.

Epidemiological aspects and risk factors of toxocariasis in a pediatric population in Sri Lanka.

This cross-sectional study, carried out over a period of 11 months, investigated the relationship between Toxocara seropositivity, socio-demographic and environmental variables in a pediatric population. Risk factors for Toxocara infection were assessed by direct interview of parent or guardian using a structured pre-tested questionnaire. Eosinophilia and presence of helminth eggs or protozoan cysts in a fecal smear were recorded. Diagnosis of Toxocara seropositivity in children was based on IgG Toxocara Microwell Serum Elisa Kits. The ELISA test was regarded as positive if the optical density was 0.3 units or above. Unadjusted and adjusted odds ratios were calculated to determine risk factors for disease. The proportion of children who were positive for Toxocara antibodies in the study population was 20%. Children being exposed to a puppy of less than 3 months at home, visiting a playground frequently, living in a poorly constructed house and dogs having access to playgrounds were significant risk factors on univariate analysis. Of these four variables, only the first three variables (OR 19, OR 4 and OR 3, respectively) remained significant risk factors on the multivariate model. Presence of eosinophilia in seropositive children was significantly higher than the seronegative group (77% vs 40%; p < 0.001). This study indicates that dogs contribute significantly to children being seropositive for toxocariasis in Sri Lanka. Implementation of public health programs specifically focused on anti-parasitic treatment of dogs is recommended.

Prevalence of G6PD deficiency in selected populations from two previously high malaria endemic areas of Sri Lanka.

Glucose-6-Phosphate Dehydrogenase (G6PD) enzyme deficiency is known to offer protection against malaria and an increased selection of mutant genes in malaria endemic regions is expected. However, anti-malarial drugs such as primaquine can cause haemolytic anaemia in persons with G6PD deficiency. We studied the extent of G6PD deficiency in selected persons attending Teaching Hospitals of Anuradhapura and Kurunegala, two previously high malaria endemic districts in Sri Lanka. A total of 2059 filter-paper blood spots collected between November 2013 and June 2014 were analysed for phenotypic G6PD deficiency using the modified WST-8/1-methoxy PMS method. Each assay was conducted with a set of controls and the colour development assessed visually as well as with a microplate reader at OD450-630nm. Overall, 142/1018 (13.95%) and 83/1041 (7.97%) were G6PD deficient in Anuradhapura and Kurunegala districts respectively. The G6PD prevalence was significantly greater in Anuradhapura when compared to Kurunegala (P<0.0001). Surprisingly, females were equally affected as males in each district: 35/313 (11.18%) males and 107/705 (15.18%) females were affected in Anuradhapura (P = 0.089); 25/313 (7.99%) males and 58/728 (7.97%) females were affected in Kurunegala (P = 0.991). Prevalence was greater among females in Anuradhapura than in Kurunegala (P<0.05), while no such difference was observed between the males (P>0.05). Severe deficiency (<10% normal) was seen among 28/1018 (2.75%) in Anuradhapura (7 males; 21 females) and 17/1041 (1.63%) in Kurunegala (7 males; 10 females). Enzyme activity between 10-30% was observed among 114/1018 (11.20%; 28 males; 86 females) in Anuradhapura while it was 66/1041 (6.34%; 18 males; 48 females) in Kurunegala. Screening and educational programmes for G6PD deficiency are warranted in these high risk areas irrespective of gender for the prevention of disease states related to this condition.