Impact of Sub-patent Malaria During Pregnancy on Birth-Weight in Odisha, India: Time-to-Event Analysis of Prospective Longitudinal Follow-Up of a Survey.

OBJECTIVE: The current study aimed to estimate prevalence of malaria infection, especially sub-patent infection, in pregnant women residing in high malaria-endemic, hard-to-reach pockets of the Indian state of Odisha; and also measure its impact on birth-weight of their new-borns. METHOD: A time-to-event analysis of prospective longitudinal follow-up study nested within a cross-sectional survey of people residing in high malaria-endemic six districts of Odisha was conducted during July-November 2019. Malaria status in pregnant mothers was categorized as malaria free; sub-patent, and patent. Hazards Ratio (HR) of low birth-weight (LBW; birth-weight < 2500 gms) was estimated in these three categories (n = 308) adjusted for residence (block), gravida, caste, age and gestational age at testing. RESULTS: 50.3% pregnant women had sub-patent malaria infection, 3.9% had patent infection. In fully adjusted model, hazards ratio of LBW was 3.76 (95% CI 1.12, 12.64, p = 0.032) in pregnant women with patent infection and 1.82 (95% CI 0.87, 3.81, p = 0.109) in women with sub-patent infection when compared to no malaria group. CONCLUSION: The study showed that half of the pregnant women in high-endemic pockets had sub-patent infection which posed deleterious influence on birth-weight of their new-borns. The study thereby flags the prevalence of sub-patent infection as a public health concern, because sub-patent infection in pregnant mothers may persist as a "silent" reservoir, with the potential to derail the malaria control program, especially when the country plans malaria elimination by 2030.

Molecular surveillance of anti-malarial drug resistance genes in Plasmodium falciparum isolates in Odisha, India.

BACKGROUND: Despite significant progress in eliminating malaria from the state of Odisha, India, the disease is still considered endemic. Artesunate plus sulfadoxine-pyrimethamine (AS + SP) has been introduced since 2010 as first-line treatment for uncomplicated Plasmodium falciparum malaria. This study aimed to investigate the prevalence of mutations associated with resistance to chloroquine (CQ), sulfadoxine-pyrimethamine (SP), and artesunate (ART) in P. falciparum parasites circulating in the state. METHODS: A total of 239 isolates of P. falciparum mono infection were collected during July 2018-November 2020 from the four different geographical regions of the state. Genomic DNA was extracted from 200 µL of venous blood and amplified using nested polymerase chain reaction. Mutations on gene associated with CQ (Pfcrt and Pfmdr1) were assessed by PCR amplification and restriction fragment length polymorphism, artemisinin (Pfk13) gene by DNA sequencing and SP (Pfdhfr and Pfdhps) genes by allele-specific polymerase chain reaction (AsPCR). RESULTS: The point mutation in Pfcrt (K76T) was detected 2.1%, in Pfmdr1 (N86Y) 3.4%, and no mutations were found in Pfkelch13 propeller domain. Prevalence of Pfdhfr, Pfdhps and Pfhdfr-Pfdhps (two locus) gene mutations were 50.43%, 47.05% and 49.79% respectively. The single, double, triple and quadruple point mutations in Pfdhfr gene was 11.2%, 8.2%, 17.2% and 3.4% while, in Pfdhps gene was 10.9%,19.5%, 9.5% and 2.7% respectively. Of the total 13 haplotypes found in Pfdhfr, 8 were detected for the first time in the state and of the total 26 haplotypes found in Pfdhps, 7 were detected for the fisrt time in the state. The linked quintuple mutation Pfdhfr (N51I-C59R-S108N)-Pfdhps (A437G-K540E) responsible for clinical failure (RIII level of resistance) of SP resistance and A16V-S108T mutation in Pfdhfr responsible for cycloguanil was absent. CONCLUSION: The study has demonstrated a low prevalence of CQ resistance alleles in the study area. Despite the absence of the Pfkelch13 mutations, high prevalence of Pfdhfr and Pfdhps point mutations undermine the efficacy of SP partner drug, thereby threatening the P. falciparum malaria treatment policy. Therefore, continuous molecular and in vivo monitoring of ACT efficacy is warranted in Odisha.