Addressing low-density malaria infections in India and other endemic part of the world-the opportune time?

Shifting from high- to low-malaria transmission accompanies a higher proportion of asymptomatic low-density malaria infections (LDMI). Currently, several endemic countries, such as India, are experiencing this shift as it is striving to eliminate malaria. LDMI is a complex concept for which there are several important questions yet unanswered on its natural history, infectiousness, epidemiology, and pathological and clinical impact. India is on the right path to eliminating malaria, but it is facing the LDMI problem. A brief discussion on the concept and definitions of LDMI is beforehand presented. Also, an exhaustive review and critical analysis of the existing literature on LDMI in malaria-endemic areas, including India, are included in this review. Finally, we opine that addressing LDMI in India is ethically and pragmatically achievable, and a pool of sine qua non conditions is required to efficiently and sustainably eliminate malaria.

Global polymorphism of Plasmodium falciparum histidine rich proteins 2/3 and impact on malaria rapid diagnostic test detection: a systematic review and meta-analysis.

BACKGROUND: This review presents an overview of field findings on sequence variation of Plasmodium falciparum histidine-rich proteins 2/3 (PfHRP2/3) for which reference types (1-24) have been identified, and its critical impact on PfHRP2-based rapid diagnostic test (RDT) detection. RESEARCH DESIGN AND METHODS: This systematic review and meta-analysis was registered with PROSPERO, CRD42022316027, and conducted as per the PRISMA guidelines, and the methodological quality of studies was assessed. RESULTS: Of the 2184 records identified, 34 studies were included mostly from Africa (47.1%) and Asia (35.3%). The reference PfHRP2 types 1, 2, 3, 6, and 7 are invariably found at proportions ≥ 80-100% in all areas with the exception of The Americas where their proportion is very low. The proteins exhibited high diversity of variants/unknown types, especially for types 1, 2, 4, and 7. Eleven major PfHRP2 epitopes were found at pooled proportion > 90%. The existing models to predict RDT detection are greatly limited by the impact of factors such as low (very low) parasitemia, RDT brand, and PfHRP3 cross-reactivity. PfHRP2 length and presence/number of a given reference repeat type/variant did not seem to impact RDT detection. CONCLUSIONS: PfHRP2/3 are highly polymorphic and current findings are insufficient, conflicting and not convincing enough to conclude on the role of PfHRP2/3 sequence polymorphism in PfHRP2-based RDT detection.

Nationwide spatiotemporal drug resistance genetic profiling from over three decades in Indian Plasmodium falciparum and Plasmodium vivax isolates.

BACKGROUND: Drug resistance is a serious impediment to efficient control and elimination of malaria in endemic areas. METHODS: This study aimed at analysing the genetic profile of molecular drug resistance in Plasmodium falciparum and Plasmodium vivax parasites from India over a ~ 30-year period (1993-2019). Blood samples of P. falciparum and/or P. vivax-infected patients were collected from 14 regions across India. Plasmodial genome was extracted and used for PCR amplification and sequencing of drug resistance genes in P. falciparum (crt, dhps, dhfr, mdr1, k13) and P. vivax (crt-o, dhps, dhfr, mdr1, k12) field isolates. RESULTS: The double mutant pfcrt SVMNT was highly predominant across the country over three decades, with restricted presence of triple mutant CVIET from Maharashtra in 2012. High rates of pfdhfr-pfdhps quadruple mutants were observed with marginal presence of "fully resistant" quintuple mutant ACIRNI-ISGEAA. Also, resistant pfdhfr and pfdhps haplotype has significantly increased in Delhi between 1994 and 2010. For pfmdr1, only 86Y and 184F mutations were present while no pfk13 mutations associated with artemisinin resistance were observed. Regarding P. vivax isolates, the pvcrt-o K10 "AAG" insertion was absent in all samples collected from Delhi in 2017. Pvdhps double mutant SGNAV was found only in Goa samples of year 2008 for the first time. The pvmdr1 908L, 958M and 1076L mutations were highly prevalent in Delhi and Haryana between 2015 and 2019 at complete fixation. One nonsynonymous novel pvk12 polymorphism was identified (K264R) in Goa. CONCLUSIONS: These findings support continuous surveillance and characterization of P. falciparum and P. vivax populations as proxy for effectiveness of anti-malarial drugs in India, especially for independent emergence of artemisinin drug resistance as recently seen in Africa.

Plasmodium cynomolgi in humans: current knowledge and future directions of an emerging zoonotic malaria parasite.

Plasmodium cynomolgi (Pcy), a simian malaria parasite, is a recent perfect example of emerging zoonotic transfer in human. This review summarizes the current knowledge on the epidemiology of natural Pcy infections in humans, mosquitoes and monkeys, along with its biological, clinical and drug sensitivity patterns. Knowledge gaps and further studies on Pcy in humans are also discussed. This parasite currently seems to be geographically limited in South-East Asia (SEA) with a global prevalence in human ranging from 0 to 1.4%. The Pcy infections were reported in local SEA populations and European travelers, and range from asymptomatic carriage to mild/moderate attacks with no evidence of pathognomonic clinical and laboratory patterns but with Pcy strain-shaped clinical differences. Geographical distribution and competence of suitable mosquito vectors and non-primate hosts, globalization, climate change, and increased intrusion of humans into the habitat of monkeys are key determinants to emergence of Pcy parasites in humans, along with its expansion outside SEA. Sensitization/information campaigns coupled with training and assessment sessions of microscopists and clinicians on Pcy are greatly needed to improve data on the epidemiology and management of human Pcy infection. There is a need for development of sensitive and specific molecular tools for individual diagnosis and epidemiological studies. The development of safe and efficient anti-hypnozoite drugs is the main therapeutic challenge for controlling human relapsing malaria parasites. Experience gained from P. knowlesi malaria, development of integrated measures and strategies-ideally with components related to human, monkeys, mosquito vectors, and environment-could be very helpful to prevent emergence of Pcy malaria in humans through disruption of transmission chain from monkeys to humans and ultimately contain its expansion in SEA and potential outbreaks in a context of malaria elimination.

Epidemiology and clinical outcomes of severe Plasmodium vivax malaria in India.

OBJECTIVES: A systematic review and meta-analysis (SR-MA) of the available Indian literature on severe vivax malaria (SVM) was undertaken. METHODS: Relevant studies in eight electronic databases were retrieved and reviewed. The preferred reporting items for systematic reviews and meta-analyses (PRISMA) guidelines were followed. The methodological quality of the studies included in the MA was assessed. RESULTS: Overall, 162 studies were included in the work. The pooled proportion of SVM was 29.3%. The main severity signs/symptoms seen in SVM were jaundice, severe thrombocytopenia (ST), multi-organ dysfunction, and severe anaemia with pooled proportion of 37.4%, 37.2%, 24.2% and 20.4%, respectively. P. falciparum was inducing 6% less ST (RR = 0.94, 95% CI 0.5-1.5, I2 = 77.87%), 10% less thrombocytopenia (RR = 0.9, 95% CI 0.7-1.1, I2 = 91.68%) and 20% less DIC (RR = 0.8, 95% CI 0.3-1.9, I2 = 0%) than P. vivax. An atypical condition like myocarditis, was most commonly observed among the studied SVM cases. The mortality rate in SVM cases ranged from 0 to 12.9% among hospital patients with P. vivax mono-infections. CONCLUSIONS: The present SR-MA provides evidence for P. vivax as the etiologic agent of severe malaria leading to deaths in few cases as seen recently in India. However, research gaps outlined here emphasise the need for further studies on SVM in pregnancy, SVM in drug resistance and correlations with cytoadherence in disease severity due to P. vivax.

First evidence of local circulation of Plasmodium ovale curtisi and reliability of a malaria rapid diagnostic test among symptomatic outpatients in Douala, Cameroon.

The performances of a commonly used Plasmodium falciparum-detecting rapid diagnostic test (RDT) were determined in symptomatic individuals living in Cameroon. Discrepancies between RDT and light microscopy (LM) results were further investigated, with a focus on non-falciparum malaria (NFM) which are still largely understudied in sub-Saharan Africa (sSA) countries. In the present study, a total of 355 individuals aged 1-65 years were enrolled in the study. Their signs/symptoms and sociodemographic characteristics were documented. The RDT reliability was evaluated using LM as gold standard method. Polymerase chain reaction (PCR) of Plasmodium 18S gene was performed for samples with discordant results between LM and RDT (i.e., RDT-/LM+, and RDT+/LM-). The PCR amplicons of NFM species were sequenced and BLASTed. The prevalence of malaria infection by LM was 95.7% (95% CI: 93.1-97.4%). The sensitivity and specificity of RDT for P. falciparum detection was 94.0% and 66.7%, respectively. By PCR assay, P. ovale curtisi (PoC) was found in 5 of the 30 discordant samples, and on sequence analysis these isolates were found to be phylogenetically closer to sequences reported from China-Myanmar border and Malaysia. This is the first report on molecular characterization of P. ovale subspecies in Cameroon. The study also outlines the good diagnostic performances of the RDT for detection of P. falciparum. Though, the presence of PoC indicated the importance of having RDTs targeting the NFM species in malaria diagnosis and treatment, which is presently limited in the country.

Artemisinin-based combination therapy (ACT) and drug resistance molecular markers: A systematic review of clinical studies from two malaria endemic regions - India and sub-Saharan Africa.

Artemisinin-based combination therapies (ACT) are currently used as a first-line malaria therapy in endemic countries worldwide. This systematic review aims at presenting the current scenario of drug resistance molecular markers, either selected or involved in treatment failures (TF) during in vivo ACT efficacy studies from sub-Saharan Africa (sSA) and India. Eight electronic databases were comprehensively used to search relevant articles and finally a total of 28 studies were included in the review, 21 from sSA and seven from India. On analysis, Artemether + lumefantrine (AL) and artesunate + sulfadoxine-pyrimethamine (AS + SP) are the main ACT in African and Indian regions with a 28-day efficacy range of 54.3-100% for AL and 63-100% for AS + SP respectively. It was observed that mutations in the Pfcrt (76T), Pfdhfr (51I, 59R, 108N), Pfdhps (437G) and Pfmdr1 (86Y, 184F, 1246Y) genes were involved in TF, which varied with respect to ACTs. Based on studies that have genotyped the Pfk13 gene, the reported TF cases, were mainly linked with mutations in genes associated with resistance to ACT partner drugs; indicating that the protection of the partner drug efficacy is crucial for maintaining the efficacy of ACT. This review reveals that ACT are largely efficacious in India and sSA despite the fact that some clinical efficacy and epidemiological studies have reported some validated mutations (i.e., 476I, 539T and 561H) in circulation in these two regions. Also, the role of PfATPase6 in ART resistance is controversial still, while P. falciparum plasmepsin 2 (Pfpm2) in piperaquine (PPQ) resistance and dihydroartemisinin (DHA) + PPQ failures is well documented in Southeast Asian countries but studied less in sSA. Hence, there is a need for continuous molecular surveillance of Pfk13 mutations for emergence of artemisinin (ART) resistance in these countries.

Rifampicin resistance among Mycobacterium tuberculosis-infected individuals using GeneXpert MTB/RIF ultra: a hospital-based study.

OBJECTIVES: To determine the prevalence and risk factors associated with rifampicin (RIF)-resistant tuberculosis using GeneXpert technology. METHODS: A cross-sectional study was conducted from April 2018 to November 2019 among tuberculosis (TB)-infected Cameroonian patients in the Littoral Region using records from patients presenting with clinically suspected or documented TB. The patients were screened for TB using GeneXpert MDR/RIF ultra. Data were documented with an ad hoc survey form and analysed with SPSS version 22. RESULTS: 153 patients were included in the study. 64.1% were males; mean age was 37.9 ± 14.7 years and median age 37 years (range: 2-82). Most patients were new cases (76.4%). Relapses accounted for 8.5% and recurrences for 2.6%. Pulmonary TB was diagnosed among 98.7% patients using mostly sputum samples (85%). The prevalence of RIF resistance was 6.7% (95% CI: 3.4%-12.7%). This prevalence was significantly higher in samples of mucus and mucopurulent aspect (P-value = 0.04). RIF-resistant M. tuberculosis strains were significantly more frequent among relapses than new cases (23.1% vs. 2.3% P-value < 0.0001). A statistically significant association was found between GeneXpert-based quantification results and type and aspect of samples. CONCLUSION: This study confirms the circulation of RIF-resistant M. tuberculosis strains in the Littoral region. There is a need for extensive studies in other parts of the country.

Green-synthesized metal nanoparticles for mosquito control: A systematic review about their toxicity on non-target organisms.

Studies capturing the high efficiency of green-synthesized metal nanoparticles (NPs) in targeting mosquito vectors of the world's main infectious diseases suggest the NPs' possible utilization as bio-insecticides. However, it is necessary to confirm that these potential bio-insecticides are not harmful to non-target organisms that are often sympatric and natural enemies of the vectors of these diseases. In this systematic review, we comprehensively analyse the content of 56 publications focused on the potentially deleterious effects of NPs on these non-target organisms. Current research on biosynthesised NPs, characterization, and impact on mosquito vectors and non-target larvivorous organisms is reviewed and critically discussed. Finally, we pinpoint some major challenges that merit future investigation. Plants (87.5%) were mainly used for synthesizing NPs in the studies. NPs were found to be spherical or mainly spherical in shape with a large distribution size. In most of the included studies, NPs showed interesting mosquitocidal activity (LC50 < 50 ppm). Some plant families (e.g., Meliaceae, Poaceae, Lamiaceae) have produced NPs with a particularly high larvicidal and pupicidal activity (LC50 < 10 ppm). Regarding non-target organisms, most of the studies concluded that NPs were safe to them, with boosted predatory activity in NP-treated milieu. In contrast, some studies reported NP-elicited adverse effects (i.e., genotoxic, nuclear, and enzymatic effects) on these non-target organisms. This review outlines the promising mosquitocidal effects of biosynthesized NPs, recognizing that NPs' potential usage is currently limited by the harm NPs are thought pose to non-target organism. It is of utmost importance to investigate green NPs to determine whether laboratory findings have applications in the real world.

Comparison of the fluorescence microscopy Cyscope® with light microscopy for malaria diagnosis in a small and active surveillance in Cameroon.

BACKGROUND: Malaria has a negative impact on the activities of companies in endemic countries especially in Cameroon. In this regard, an increasingly growing number of companies have started to include management of malarious patients in their health policies. In the present study, we will evaluate the diagnostic performances of a fluorescence microscopy (FM), Cyscope® microscope, in the detection of malaria parasites. METHODS: A cross-sectional study was conducted among employees of two companies of the town of Douala on 21 and 22 March 2017. Sociodemographic information of employees was collected using a questionnaire form. Blood samples of ~ 10 µL were collected by venipuncture for the diagnosis of malaria using FM and light microscopy (LM). Performances of FM with respect to sensitivity (Se), specificity (Sp), positive and negative predictive values (PPV and NPV), positive and negative likelihood rates (PLR and NLR), accuracy, reliability, and Kappa index were calculated using LM as gold standard. RESULTS: In total, 442 employees, aged 37.8 ± 9.7 years old on average, were included in the study. Prevalence of malaria using FM and LM was 39.2% and 17%, respectively (p < 0.01). Plasmodium falciparum and P. vivax were the two species involved in malaria infection cases. In terms of developmental stages, 68%, 45.3%, and 1.3% of employees carried gametocytes, trophozoites, and schizonts, respectively. Findings on diagnostic performances of FM were as follows: Se = 84%, Sp = 69.95%, PPV = 63.58%, NPV = 95.5%, accuracy = 89.36%, and reliability = 53.95%. Sensitivity of Cyscope® microscope increased as a function of parasitemia with values ranging from 76.92% at parasitemia between 1 and 500 parasites/µL to 91.11% at parasitemia between 501 and 5000 parasites/µL. The geometric mean parasite density was1850 parasites per µL of blood (range 1600-40,000), and most of employees (60.8%) had moderate parasitemia. The performances of FM were similar between febrile and afebrile patients. CONCLUSIONS: This study showed good performances of Cyscope® microscope and outlines that this diagnostic tool could be used in management of malaria at workplace.

Tuberculosis Chemotherapy Outcome in the Littoral Region of Cameroon: A Meta-analysis of Treatment Success Rate between 2014 and 2016.

BACKGROUND: Tuberculosis (TB) is a public health concern, especially in resource-constrained countries like Cameroon. TB drug resistance is a major obstacle to control and prevent. DESIGN: Data from 2014 to 2016 on the outcome of anti-TB treatment in the Littoral Region were reviewed manually and analysed using the meta-analysis concept. The treatment success rates (TSR) were the primary outcome used for this study. The heterogeneity statistics (I 2) was computed to orientate the choice of the best statistical model (binary fixed effect or random) to compute pooled value of TSR. RESULTS: Using an intention-to-treat analysis, the pooled proportions of HIV-uninfected TB patients successfully cured from TB were low and slightly decreased by 1% between 2014 and 2016. Regarding HIV-infected TB patients, pooled values of TSR were lower than those of their HIV-negative counterparts with values ranging from 71% (95% CI: 63%-83%; I 2 = 71.16%) in 2014 to 68% (95% CI: 58%-79%; I 2 = 70.97%) in 2016. In addition, no heterogeneity was found in three years (I 2 = 0.0%; P value = 1). These cure rates were strongly and negatively correlated with the rates of patients lost to follow-up regardless of the year. In HIV-infected patients, the pooled values of ITT analysis-based treatment success rates were 73% (χ 2 = 13.92, P value = 0.0002), 71% (χ 2 = 7.26, P value = 0.007), and 68% (χ 2 = 8.02, P value = 0.004), respectively. The coverage rates with cotrimoxazole (CTX) gradually increased over year ranging from 78.90% in 2014 to 94.17% in 2016, similar to the coverage rate for ARV therapy that was 60.06% in 2014 against 90% in 2016. A positive and statistically significant correlation was found between the success of the anti-TB therapy in HIV-infected patients and coverage rates with CTX and ARV. CONCLUSION: An improvement in the reduction of percentage of lost to follow-up and coverage with CTX and ARV therapy could greatly increase chances to efficiently control TB in Cameroon.

A systematic review on anti-malarial drug discovery and antiplasmodial potential of green synthesis mediated metal nanoparticles: overview, challenges and future perspectives.

BACKGROUND: The recent emergence in Southeast Asia of artemisinin resistance poses major threats to malaria control and elimination globally. Green nanotechnologies can constitute interesting tools for discovering anti-malarial medicines. This systematic review focused on the green synthesis of metal nanoparticles as potential source of new antiplasmodial drugs. METHODS: Seven electronic database were used following the Preferred Reporting Items for Systematic Reviews and Meta-Analyses (PRISMA) guidelines. RESULTS: A total of 17 papers were included in the systematic review. 82.4% of the studies used plant leaves to produce nanoparticles (NPs) while three studies used microorganisms, including bacteria and fungi. Silver was the main metal precursor for the synthesis of NPs. The majority of studies obtained nanoparticles spherical in shape, with sizes ranging between 4 and 65 nm, and reported no or little cytotoxic effect of the NPs. Results based on 50% inhibitory concentration (IC50) varied between studies but, in general, could be divided into three NP categories; (i) those more effective than positive controls, (ii) those more effective than corresponding plant extracts and, (iii) those less effective than the positive controls or plant extracts. CONCLUSIONS: This study highlights the high antiplasmodial potential of green-synthesized metal nanoparticles thereby underscoring the possibility to find and develop new anti-malarial drugs based on green synthesis approaches. However, the review also highlights the need for extensive in vitro and in vivo studies to confirm their safety in humans and the elucidation of the mechanism of action.