Field site soil aquifer treatment shows enhanced wastewater quality: Evidence from vadose zone hydro-geophysical observations.

Soil aquifer treatment (SAT) is an emerging, nature-based, economically viable wastewater treatment solution. Currently, most SAT experiments are done at the laboratory scale, which cannot generate the same conditions as natural field sites and limits the understanding of treatment efficiency. The current study carried out in situ SAT experiments in the Musi River basin in India, where wastewater irrigation is a common practice. SAT efficiency was determined using an integrated approach, including electrical resistivity tomography (ERT) surveys, soil investigations (grain size, permeability, and moisture measurements), and biochemical characterization of raw and SAT treated wastewater. The ERT scans of SAT column show lower order electrical resistivity 10-30 Ω-m with enhanced chargeability >5-6 mV/V attributed to the vadose zone, characterized by clay-rich soil and sandy soil up to 5-6 m depth. The increase in sand percentage (>70%) below 140-160 cm depth corroborates with the high moisture content (23.5%). The vadose zone permeability (K) 1.58 m/day and discharge (Q) 38.19 m3/day is used to determine the pollutants reduction efficiency of SAT column. Hydrogeological and biogeochemical observations reveal that the improved dissolved oxygen from <1.0 to 5-6 mg/L in the vadose zone catalyzes the oxidation of organic matter resulting in the reduction of BOD and COD up to 92% and 97%, respectively, and denitrification reducing NO3-- (0.55 kg/day). In addition, the precipitation and adsorption by kaolinite clay prompted the reduction of PO42- (0.26 kg/day). Furthermore, the oxic-vadose zone could not support the growth of coliforms and faecal coliforms, and the reduction observed was up to 99.99% in the SAT production well. Overall, the results indicated a positive outcome with SAT efficiency and framed the SAT sitting criteria for different geological environments.

Hydrogeochemical and mixing processes controlling groundwater chemistry in a wastewater irrigated agricultural system of India.

In many parts of the world, wastewater irrigation has become a common practice because of freshwater scarcity and to increase resource reuse efficiency. Wastewater irrigation has positive impacts on livelihoods and at the same time, it has adverse impacts related to environmental pollution. Hydrochemical processes and groundwater behaviour need to be analyzed for a thorough understanding of the geochemical evolution in the wastewater irrigated systems. The current study focuses on a micro-watershed in the peri-urban Hyderabad of India, where farmers practice intensive wastewater irrigation. To evaluate the major factors that control groundwater geochemical processes, we analyzed the chemical composition of the wastewater used for irrigation and groundwater samples on a monthly basis for one hydrological year. The groundwater samples were collected in three settings of the watershed: wastewater irrigated area, groundwater irrigated area and upstream peri-urban area. The collected groundwater and wastewater samples were analyzed for major anions, cations and nutrients. We systematically investigated the anthropogenic influences and hydrogeochemical processes such as cation exchange, precipitation and dissolution of minerals using saturated indices, and freshwater-wastewater mixtures at the aquifer interface. Saturation indices of halite, gypsum and fluorite are exhibiting mineral dissolution and calcite and dolomite display mineral precipitation. Overall, the results suggest that the groundwater geochemistry of the watershed is largely controlled by long-term wastewater irrigation, local rainfall patterns and water-rock interactions. The study results can provide the basis for local decision-makers to develop sustainable groundwater management strategies and to control the aquifer pollution influenced by wastewater irrigation.

Hydro-geophysical characterization and performance evaluation of natural wetlands in a semi-arid wastewater irrigated landscape.

Natural wetlands are green infrastructure systems that are energy-efficient for wastewater treatment and can be found in diverse geo-environmental settings around the world. Their structure and functions, which defines the treatment efficiencies are highly varied. Wetlands over shallow bedrock and geological lineaments (weak zones) have been known to contribute to groundwater contamination. However, not many studies have been performed to understand the structure in different geological settings to identify the efficiency determining factors. Therefore, it is important to investigate the geological suitability of the natural wetlands. We examined wastewater fed natural wetlands in diverse geological settings aiming at studying the depth, geo-stability, bio-chemical interactions, and hydrogeological attributes that improve the wastewater quality, within the Musi River basin, India. The integrated geophysical scans encompassing electrical resistivity tomography (ERT), hydrogeological test, bathymetric study and hydro-chemical analysis were carried out to explore the physical structure and hydro-dynamic processes in the wetlands. ERT investigations showed that, the depth to bedrock up to 20-25 m devoid of geo-fractures (lineaments) indicated the effective depth of saturated zone as a passable scope for potential bio-chemical interactions, implying the proportionality of the deep seated (deep bedrock) wetland to the pollutant removal efficiency. The lower order of electrical resistivity range 10-35 Ωm and hydraulic conductivity 2.938 md-1 acquired for saturated weathered zone were found catalyzing the bioremediation, sedimentation, adsorption, redox reactions and ion exchange processes. It caused the deep seated wetland removing nitrate 194.34 kgd-1 (97.18%); sulphate 333.75 kgd-1 (77.70%); phosphate 9.66 kgd-1 (82.53%); microbes 99.99%, BOD 80%, and COD 80% load with discharge 1408 m3d-1 of treated wastewater. Further, the strategies for designating the natural wetlands as wastewater treatment systems are also discussed in this paper.

Spatio-temporal distribution and chemical characterization of groundwater quality of a wastewater irrigated system: A case study.

Wastewater irrigation is a common livelihood practice in many parts of the developing world. With the continuous irrigation supply, groundwater systems in these regions perceive adverse impacts due to inadequate infrastructure to treat the wastewater. The current study area, Musi River irrigation system, is one such case study located in the peri-urban Hyderabad of South India. The Musi River water, which is used for irrigation, is composed of untreated and secondary treated wastewater from Hyderabad city. Kachiwani Singaram micro-watershed in the peri-urban Hyderabad is practicing wastewater irrigation for the last 40 years. The current quality of (untreated) wastewater used for irrigation is expected to have adverse impacts on the local aquifers, but detailed investigations are lacking. To elucidate the groundwater quality dynamics and seasonality of the wastewater irrigation impacts on the peri-urban agricultural system, we analyzed the groundwater quality on a monthly basis for one hydrological year in the wastewater and groundwater irrigated areas, which exist next to each other. The spatio-temporal variability of groundwater quality in the watershed was analyzed with respect to wastewater irrigation and seasonality using multivariate statistical analysis, multi-way modeling and self-organizing maps. This study indicates the significance of combining various statistical techniques for detailed evaluation of the groundwater processes in a wastewater irrigated agricultural system. The results suggest that concentrations of the major ionic substances increase after the monsoon season, especially in wastewater irrigated areas. Multi-way modeling identified the major polluted groundwaters to come from the wastewater irrigated parts of the watershed. Clusters of chemical variables identified by using self-organizing maps indicate that groundwater pollution is highly impacted by mineral interactions and long-term wastewater irrigation. The study recommends regular monitoring of water resources and development of sustainable management strategies to mitigate the aquifer pollution in wastewater irrigation systems.

Natural treatment system models for wastewater management: a study from Hyderabad, India.

Wastewater generated on a global scale has become a significant source of water resources which necessitates appropriate management strategies. However, the complexities associated with wastewater are lack of economically viable treatment systems, especially in low- and middle-income countries. While many types of treatment systems are needed to serve the various local issues, we propose natural treatment systems (NTS) such as natural wetlands that are eco-friendly, cost-effective, and can be jointly driven by public bodies and communities. In order for it to be part of wastewater management, this study explores the NTS potential for removal of pollutants, cost-effectiveness, and reuse options for the 1.20 million m3/day of wastewater generated in Hyderabad, India. The pilot study includes hydro-geophysical characterization of natural wetland to determine pollutant removal efficiency and its effective utilization for treated wastewater in the peri-urban habitat. The results show the removal of organic content (76-78%), nutrients (77-97%), and microbes (99.5-99.9%) from the wetland-treated wastewater and its suitability for agriculture applications. Furthermore, the wetland efficiency integrated with engineered interventions led to the development of NTS models with different application scenarios: (i) constructed wetlands, (ii) minimized community wetlands, and (iii) single outlet system, suitable for urban, peri-urban and rural areas, respectively.

Combined Effects of Pesticides and Trematode Infections on Hourglass Tree Frog Polypedates cruciger.

The impact of widespread and common environmental factors, such as chemical contaminants, on infectious disease risk in amphibians is particularly important because both chemical contaminants and infectious disease have been implicated in worldwide amphibian declines. Here we report on the lone and combined effects of exposure to parasitic cercariae (larval stage) of the digenetic trematode, Acanthostomum burminis, and four commonly used pesticides (insecticides: chlorpyrifos, dimethoate; herbicides: glyphosate, propanil) at ecologically relevant concentrations on the survival, growth, and development of the common hourglass tree frog, Polypedates cruciger Blyth 1852. There was no evidence of any pesticide-induced mortality on cercariae because all the cercariae successfully penetrated each tadpole host regardless of pesticide treatment. In isolation, both cercarial and pesticide exposure significantly decreased frog survival, development, and growth, and increased developmental malformations, such as scoliosis, kyphosis, and also edema and skin ulcers. The combination of cercariae and pesticides generally posed greater risk to frogs than either factor alone by decreasing survival or growth or increasing time to metamorphosis or malformations. The exception was that lone exposure to chlorpyrifos had higher mortality without than with cercariae. Consistent with mathematical models that suggest that stress should increase the impact of generalist parasites, the weight of the evidence from the field and laboratory suggests that ecologically relevant concentrations of agrochemicals generally increase the threat that trematodes pose to amphibians, highlighting the importance of elucidating interactions between anthropogenic activities and infectious disease in taxa of conservation concern.

Multiple Origins of Mutations in the mdr1 Gene--A Putative Marker of Chloroquine Resistance in P. vivax.

BACKGROUND: Chloroquine combined with primaquine has been the recommended antimalarial treatment of Plasmodium vivax malaria infections for six decades but the efficacy of this treatment regimen is threatened by chloroquine resistance (CQR). Single nucleotide polymorphisms (SNPs) in the multidrug resistance gene, Pvmdr1 are putative determinants of CQR but the extent of their emergence at population level remains to be explored. OBJECTIVE: In this study we describe the prevalence of SNPs in the Pvmdr1 among samples collected in seven P. vivax endemic countries and we looked for molecular evidence of drug selection by characterising polymorphism at microsatellite (MS) loci flanking the Pvmdr1 gene. METHODS: We examined the prevalence of SNPs in the Pvmdr1 gene among 267 samples collected from Pakistan, Afghanistan, Sri Lanka, Nepal, Sudan, São Tomé and Ecuador. We measured and diversity in four microsatellite (MS) markers flanking the Pvmdr1 gene to look evidence of selection on mutant alleles. RESULTS: SNP polymorphism in the Pvmdr1 gene was largely confined to codons T958M, Y976F and F1076L. Only 2.4% of samples were wildtype at all three codons (TYF, n = 5), 13.3% (n = 28) of the samples were single mutant MYF, 63.0% of samples (n = 133) were double mutant MYL, and 21.3% (n = 45) were triple mutant MFL. Clear geographic differences in the prevalence of these Pvmdr mutation combinations were observed. Significant linkage disequilibrium (LD) between Pvmdr1 and MS alleles was found in populations sampled in Ecuador, Nepal and Sri Lanka, while significant LD between Pvmdr1 and the combined 4 MS locus haplotype was only seen in Ecuador and Sri Lanka. When combining the 5 loci, high level diversity, measured as expected heterozygosity (He), was seen in the complete sample set (He = 0.99), while He estimates for individual loci ranged from 0.00-0.93. Although Pvmdr1 haplotypes were not consistently associated with specific flanking MS alleles, there was significant differentiation between geographic sites which could indicate directional selection through local drug pressure. CONCLUSIONS: Our observations suggest that Pvmdr1 mutations emerged independently on multiple occasions even within the same population. In Sri Lanka population analysis at multiple sites showed evidence of local selection and geographical dispersal of Pvmdr1 mutations between sites.

Global and local genetic diversity at two microsatellite loci in Plasmodium vivax parasites from Asia, Africa and South America.

BACKGROUND: Even though Plasmodium vivax has the widest worldwide distribution of the human malaria species and imposes a serious impact on global public health, the investigation of genetic diversity in this species has been limited in comparison to Plasmodium falciparum. Markers of genetic diversity are vital to the evaluation of drug and vaccine efficacy, tracking of P. vivax outbreaks, and assessing geographical differentiation between parasite populations. METHODS: The genetic diversity of eight P. vivax populations (n=543) was investigated by using two microsatellites (MS), m1501 and m3502, chosen because of their seven and eight base-pair (bp) repeat lengths, respectively. These were compared with published data of the same loci from six other P. vivax populations. RESULTS: In total, 1,440 P. vivax samples from 14 countries on three continents were compared. There was highest heterozygosity within Asian populations, where expected heterozygosity (He) was 0.92-0.98, and alleles with a high repeat number were more common. Pairwise FST revealed significant differentiation between most P. vivax populations, with the highest divergence found between Asian and South American populations, yet the majority of the diversity (~89%) was found to exist within rather than between populations. CONCLUSIONS: The MS markers used were informative in both global and local P. vivax population comparisons and their seven and eight bp repeat length facilitated population comparison using data from independent studies. A complex spatial pattern of MS polymorphisms among global P. vivax populations was observed which has potential utility in future epidemiological studies of the P. vivax parasite.

Generalized seasonal autoregressive integrated moving average models for count data with application to malaria time series with low case numbers.

INTRODUCTION: With the renewed drive towards malaria elimination, there is a need for improved surveillance tools. While time series analysis is an important tool for surveillance, prediction and for measuring interventions' impact, approximations by commonly used Gaussian methods are prone to inaccuracies when case counts are low. Therefore, statistical methods appropriate for count data are required, especially during "consolidation" and "pre-elimination" phases. METHODS: Generalized autoregressive moving average (GARMA) models were extended to generalized seasonal autoregressive integrated moving average (GSARIMA) models for parsimonious observation-driven modelling of non Gaussian, non stationary and/or seasonal time series of count data. The models were applied to monthly malaria case time series in a district in Sri Lanka, where malaria has decreased dramatically in recent years. RESULTS: The malaria series showed long-term changes in the mean, unstable variance and seasonality. After fitting negative-binomial Bayesian models, both a GSARIMA and a GARIMA deterministic seasonality model were selected based on different criteria. Posterior predictive distributions indicated that negative-binomial models provided better predictions than Gaussian models, especially when counts were low. The G(S)ARIMA models were able to capture the autocorrelation in the series. CONCLUSIONS: G(S)ARIMA models may be particularly useful in the drive towards malaria elimination, since episode count series are often seasonal and non-stationary, especially when control is increased. Although building and fitting GSARIMA models is laborious, they may provide more realistic prediction distributions than do Gaussian methods and may be more suitable when counts are low.

Malformations and mortality in the Asian Common Toad induced by exposure to pleurolophocercous cercariae (Trematoda: Cryptogonimidae).

Malformations and increased mortality due to infection by the digenetic trematode, Riberioa ondatrae have been reported for many species of amphibians. Severe malformations have also been reported in the Common Hourglass Tree Frog, Polypedates cruciger induced by pleurolophocercous cercariae in Sri Lanka in addition to the changes in the behaviour, development and survival of the host. We exposed pre-limb bud stage tadpoles (Gosner stages 25-26) of the Asian Common Toad, Duttaphrynus melanostictus to the same pleurolophocercous type cercariae under laboratory conditions. Molecular and morphological identification showed that these cercariae belonged Acanthostomum burminis infecting freshwater snakes as definitive hosts. These cercariae induced malformations (27.8%) and reduced survival to metamorphosis (53.8%). The magnitude of the effects increased with the dose of cercariae. Types of malformations were mainly axial, such as scoliosis and kyphosis. Severe limb malformations such as extra or missing limbs as reported for amphibians exposed to R. ondatrae were not observed in the D. melanostictus. Same authors reported a higher percentage of malformations previously when P. cruciger was exposed to the cercariae A. burminis compared to D. melanostictus. However, tadpoles of D. melanostictus, which are smaller compared to those of P. cruciger, experienced higher mortality than P. cruciger tadpoles. Trematode induced malformations and mortality in amphibians are highly variable and depend on multiple factors such as host species differences such as resistance to infection and tolerance, life-history characteristics such as size at metamorphosis and length of the metamorphosis period, and other factors such as size of the amphibian at the time of trematode exposure.

Analysis of polymorphisms in the merozoite surface protein-3α gene and two microsatellite loci in Sri Lankan Plasmodium vivax: evidence of population substructure in Sri Lanka.

The geographical distribution of genetic variation in Plasmodium vivax samples (N = 386) from nine districts across Sri Lanka is described using three markers; the P. vivax merozoite surface protein-3α (Pvmsp-3α) gene, and the two microsatellites m1501 and m3502. At Pvmsp-3α, 11 alleles were found with an expected heterozygosity (H(e)) of 0.81, whereas at m1501 and m3502, 24 alleles (H(e) = 0.85) and 8 alleles (H(e) = 0.74) were detected, respectively. Overall, 95 unique three locus genotypes were detected among the 279 samples positive at all three loci (H(e) = 0.95). Calculating the pairwise fixation index (F(ST)) revealed statistically significant population structure. The presence of identical 2-loci microsatellite genotypes in a significant proportion of samples revealed local clusters of closely related isolates contributing to strong linkage disequilibrium between marker alleles. The results show evidence of high genetic diversity and possible population substructure of P. vivax populations in Sri Lanka.

Distribution pattern of Plasmodium falciparum chloroquine transporter (pfcrt) gene haplotypes in Sri Lanka 1996-2006.

Widespread antimalarial resistance has been a barrier to malaria elimination efforts in Sri Lanka. Analysis of genetic markers in historic parasites may uncover trends in the spread of resistance. We examined the frequency of Plasmodium falciparum chloroquine transporter (pfcrt; codons 72-76) haplotypes in Sri Lanka in 1996-1998 and 2004-2006 using a high-resolution melting assay. Among 59 samples from 1996 to 1998, we detected the SVMNT (86%), CVMNK (10%), and CVIET (2%) haplotypes, with a positive trend in SVMNT and a negative trend in CVMNK frequency (P = 0.004) over time. Among 24 samples from 2004 to 2006, we observed only the SVMNT haplotype. This finding indicates selection for the SVMNT haplotype over time and its possible fixation in the population.

Uncertain etiologies of proteinuric-chronic kidney disease in rural Sri Lanka.

The global prevalence of chronic kidney disease (CKD) of uncertain etiology may be underreported. Community-level epidemiological studies are few due to the lack of national registries and poor focus on the reporting of non-communicable diseases. Here we describe the prevalence of proteinuric-CKD and disease characteristics of three rural populations in the North Central, Central, and Southern Provinces of Sri Lanka. Patients were selected using the random cluster sampling method and those older than 19 years of age were screened for persistent dipstick proteinuria. The prevalence of proteinuric-CKD in the Medawachchiya region (North Central) was 130 of 2600 patients, 68 of 709 patients in the Yatinuwara region (Central), and 66 of 2844 patients in the Hambantota region (Southern). The mean ages of these patients with CKD ranged from 44 to 52 years. Diabetes and long-standing hypertension were the main risk factors of CKD in the Yatinuwara and Hambantota regions. Age, exceeding 60 years, and farming were strongly associated with proteinuric-CKD in the Medawachchiya region; however, major risk factors were uncertain in 87% of these patients. Of these patients, 26 underwent renal biopsy; histology indicated tubulointerstitial disease. Thus, proteinuric-CKD of uncertain etiology is prevalent in the North Central Province of Sri Lanka. In contrast, known risk factors were associated with CKD in the Central and Southern Provinces.

Geographic structure of Plasmodium vivax: microsatellite analysis of parasite populations from Sri Lanka, Myanmar, and Ethiopia.

Genetic diversity and population structure of Plasmodium vivax parasites can predict the origin and spread of novel variants within a population enabling population specific malaria control measures. We analyzed the genetic diversity and population structure of 425 P. vivax isolates from Sri Lanka, Myanmar, and Ethiopia using 12 trinucleotide and tetranucleotide microsatellite markers. All three parasite populations were highly polymorphic with 3-44 alleles per locus. Approximately 65% were multiple-clone infections. Mean genetic diversity (H(E)) was 0.7517 in Ethiopia, 0.8450 in Myanmar, and 0.8610 in Sri Lanka. Significant linkage disequilibrium was maintained. Population structure showed two clusters (Asian and African) according to geography and ancestry. Strong clustering of outbreak isolates from Sri Lanka and Ethiopia was observed. Predictive power of ancestry using two-thirds of the isolates as a model identified 78.2% of isolates accurately as being African or Asian. Microsatellite analysis is a useful tool for mapping short-term outbreaks of malaria and for predicting ancestry.

Pre-elimination stage of malaria in Sri Lanka: assessing the level of hidden parasites in the population.

BACKGROUND: With the dramatic drop in the transmission of malaria in Sri Lanka in recent years, the country entered the malaria pre-elimination stage in 2008. Assessing the community prevalence of hidden malaria parasites following several years of extremely low transmission is central to the process of complete elimination. The existence of a parasite reservoir in a population free from clinical manifestations, would influence the strategy for surveillance and control towards complete elimination. METHODS: The prevalence of hidden parasite reservoirs in two historically malaria endemic districts, Anuradhapura and Kurunegala, previously considered as high malaria transmission areas in Sri Lanka, where peaks of transmission follow the rainy seasons was assessed. Blood samples of non-febrile individuals aged five to 55 years were collected from randomly selected areas in the two districts at community level and a questionnaire was used to collect demographic information and movement of the participants. A simple, highly sensitive nested PCR was carried out to detect both Plasmodium falciparum and Plasmodium vivax, simultaneously. RESULTS: In total, 3,023 individuals from 101 villages participated from both districts comprising mostly adults between the ages 19-55 years. Out of these, only about 1.4% of them (n = 19) could recall having had malaria during the past five years. Analysis of a subset of samples (n = 1322) from the two districts using PCR showed that none of the participants had hidden parasites. DISCUSSION: A reservoir of hidden parasites is unlikely to be a major concern or a barrier to the ongoing malaria elimination efforts in Sri Lanka. However, as very low numbers of indigenous cases are still recorded, an island-wide assessment and in particular, continued alertness and follow up action are still needed. The findings of this study indicate that any future assessments should be based on an adaptive sampling approach, involving prompt sampling of all subjects within a specified radius, whenever a malaria case is identified in a given focus.

Malaria seasonality and rainfall seasonality in Sri Lanka are correlated in space.

Correlation in space between seasonality of malaria and seasonality of rainfall was studied in Sri Lanka. A simple seasonality index was developed by making use of the bimodal seasonality of both malaria and rainfall. The malaria seasonality index was regressed against the rainfall seasonality index taking spatial autocorrelation into account. Despite the presence of spatial autocorrelation, the coefficient for the rainfall seasonality index in explaining the malaria seasonality index was found to be significant. The results suggest that rainfall is an important driver of malaria seasonality.

Epochal changes in the association between malaria epidemics and El Niño in Sri Lanka.

BACKGROUND: El Niño events were suggested as a potential predictor for malaria epidemics in Sri Lanka based on the coincidence of nine out of 16 epidemics with El Niño events from 1870 to 1945. Here the potential for the use of El Niño predictions to anticipate epidemics was examined using enhanced climatic and epidemiological data from 1870 to 2000. METHODS: The epidemics start years were identified by the National Malaria Control Programme and verified against epidemiological records for consistency. Monthly average rainfall climatologies were estimated for epidemic and non-epidemic years; as well El Niño, Neutral and La Niña climatic phases. The relationship between El Niño indices and epidemics was examined to identify 'epochs' of consistent association. The statistical significance of the association between El Niño and epidemics for different epochs was characterized. The changes in the rainfall-El Niño relationships over the decade were examined using running windowed correlations. The anomalies in rainfall climatology during El Niño events for different epochs were compared. RESULTS: The relationship between El Niño and epidemics from 1870 to 1927 was confirmed. The anomalies in monthly average rainfall during El Niño events resembled the anomalies in monthly average rainfall during epidemics during this period. However, the relationship between El Niño and epidemics broke down from 1928 to 1980. Of the three epidemics in these six decades, only one coincided with an El Niño. Not only did this relationship breakdown but epidemics were more likely to occur in periods with a La Niña tendency. After 1980, three of four epidemics coincided with El Niño. CONCLUSION: The breakdown of the association between El Niño and epidemics after 1928 is likely due to an epochal change in the El Niño-rainfall relationship in Sri Lanka around the 1930's. It is unlikely that this breakdown is due to the insecticide spraying programme that began in 1945 since the breakdown started in 1928. Nor does it explain the occurrence of epidemics during La Niña phase from 1928 to 1980. Although there has been renewed coincidence with El Niño after 1980, this record is too short for establishing a reliable relationship.

Models for short term malaria prediction in Sri Lanka.

BACKGROUND: Malaria in Sri Lanka is unstable and fluctuates in intensity both spatially and temporally. Although the case counts are dwindling at present, given the past history of resurgence of outbreaks despite effective control measures, the control programmes have to stay prepared. The availability of long time series of monitored/diagnosed malaria cases allows for the study of forecasting models, with an aim to developing a forecasting system which could assist in the efficient allocation of resources for malaria control. METHODS: Exponentially weighted moving average models, autoregressive integrated moving average (ARIMA) models with seasonal components, and seasonal multiplicative autoregressive integrated moving average (SARIMA) models were compared on monthly time series of district malaria cases for their ability to predict the number of malaria cases one to four months ahead. The addition of covariates such as the number of malaria cases in neighbouring districts or rainfall were assessed for their ability to improve prediction of selected (seasonal) ARIMA models. RESULTS: The best model for forecasting and the forecasting error varied strongly among the districts. The addition of rainfall as a covariate improved prediction of selected (seasonal) ARIMA models modestly in some districts but worsened prediction in other districts. Improvement by adding rainfall was more frequent at larger forecasting horizons. CONCLUSION: Heterogeneity of patterns of malaria in Sri Lanka requires regionally specific prediction models. Prediction error was large at a minimum of 22% (for one of the districts) for one month ahead predictions. The modest improvement made in short term prediction by adding rainfall as a covariate to these prediction models may not be sufficient to merit investing in a forecasting system for which rainfall data are routinely processed.

Temporal correlation between malaria and rainfall in Sri Lanka.

BACKGROUND: Rainfall data have potential use for malaria prediction. However, the relationship between rainfall and the number of malaria cases is indirect and complex. METHODS: The statistical relationships between monthly malaria case count data series and monthly mean rainfall series (extracted from interpolated station data) over the period 1972 - 2005 in districts in Sri Lanka was explored in four analyses: cross-correlation; cross-correlation with pre-whitening; inter-annual; and seasonal inter-annual regression. RESULTS: For most districts, strong positive correlations were found for malaria time series lagging zero to three months behind rainfall, and negative correlations were found for malaria time series lagging four to nine months behind rainfall. However, analysis with pre-whitening showed that most of these correlations were spurious. Only for a few districts, weak positive (at lags zero and one) or weak negative (at lags two to six) correlations were found in pre-whitened series. Inter-annual analysis showed strong negative correlations between malaria and rainfall for a group of districts in the centre-west of the country. Seasonal inter-annual analysis showed that the effect of rainfall on malaria varied according to the season and geography. CONCLUSION: Seasonally varying effects of rainfall on malaria case counts may explain weak overall cross-correlations found in pre-whitened series, and should be taken into account in malaria predictive models making use of rainfall as a covariate.

Island-wide diversity in single nucleotide polymorphisms of the Plasmodium vivax dihydrofolate reductase and dihydropteroate synthetase genes in Sri Lanka.

BACKGROUND: Single nucleotide polymorphisms (SNPs) in the Plasmodium vivax dihydrofolate reductase (Pfdhfr) and dihydropteroate synthetase (Pvdhps) genes cause parasite resistance to the antifolate drug combination, sulphadoxine/pyrimethamine (SP). Monitoring these SNPs provide insights into the level of drug pressure caused by SP use and presumably other antifolate drugs. In Sri Lanka, chloroquine (CQ) with primaquine (PQ) and SP with PQ is used as first and second line treatment, respectively, against uncomplicated Plasmodium falciparum and/or P. vivax infections. CQ/PQ is still efficacious against P. vivax infections, thus SP is rarely used and it is assumed that the prevalence of SNPs related to P. vivax SP resistance is low. However, this has not been assessed in Sri Lanka as in most other parts of Asia. This study describes the prevalence and distribution of SNPs related to P. vivax SP resistance across Sri Lanka. SUBJECTS AND METHODS: P. vivax-positive samples were collected from subjects presenting at government health facilities across nine of the major malaria endemic districts on the island. The samples were analysed for SNPs/haplotypes at codon 57, 58, 61 and 117 of the Pvdhfr gene and 383, 553 and 585 of the Pvdhps gene by applying PCR followed by a hybridization step using sequence specific oligonucleotide probes (SSOPs) in an ELISA format. RESULTS: In the study period, the government of Sri Lanka recorded 2,149 P. vivax cases from the nine districts out of which, 454 (21.1%) blood samples were obtained. Pvdhfr haplotypes could be constructed for 373 of these. The FSTS wild-haplotype was represented in 257 samples (68.9%), the double mutant LRTS haplotype was the most frequently observed mutant (24.4%) while the triple mutation (LRTN) was only identified once. Except for two samples of the single mutated Pvdhps GAV haplotype, the remaining samples were wildtype. Geographical differences were apparent, notably a significantly higher frequency of mutant Pvdhfr haplotypes was observed in the Northern districts. CONCLUSION: Since SP is rarely used in Sri Lanka, the high frequency and diversity of Pvdhfr mutations was unexpected indicating the emergence of drug resistant parasites despite a low level of SP drug pressure.