An assay for the identification of Plasmodium simium infection for diagnosis of zoonotic malaria in the Brazilian Atlantic Forest

Alvarenga, Denise Anete Madureira de; Culleton, Richard; Pina-Costa, Anielle de; Rodrigues, Danielle Fonseca; Bianco, Cesare; Silva, Sidnei G.; Nunes, Ana Júlia Dutra; Souza, Júlio César de; Hirano, Zelinda Maria Braga; Moreira, Sílvia Bahadian; Pissinatti, Alcides; Abreu, Filipe Vieira Santos de; Areas, André Luiz Lisboa; Lourenço‐de‐Oliveira, Ricardo; Zalis, Mariano Gustavo; Ferreira-da-Cruz, Maria de Fátima; Brasil, Patrícia; Daniel-Ribeiro, Cláudio Tadeu; Brito, Cristiana Ferreira Alves de

doi:10.1038/s41598-017-18216-x

Cited by 32 publications

(52 citation statements)

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“…Following a malaria outbreak in the Atlantic Forest of Rio de Janeiro in 2015/2016, it was shown that these infections were caused by the non-human primate malaria parasite P. simium 1 . DNA samples collected both from humans and non-human primates (NHPs) in the same region shared identical mitochondrial genome sequences, distinct from P. vivax isolates from anywhere in the world and identical to that of a P. simium parasite isolated from a monkey in the same region in 1966, and to all isolates of P. simium recovered from NHPs since 13,14 .…”

Section: Introductionmentioning

confidence: 99%

The genome of the zoonotic malaria parasite Plasmodium simium reveals adaptations to host-switching

Mourier

Alvarenga

Kaushik

et al. 2019

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Oxford, Headington, Oxford, OX3 9DU, UK 41 42 §) Contributed equally, arranged alphabetically 43 †) Corresponding authors 44 45 46 Summary 47 48 Plasmodium simium, a malaria parasite of non-human primates in the Atlantic forest region of 49Brazil was recently shown to cause zoonotic infection in humans in the region. Phylogenetic 50 analyses based on the whole genome sequences of six P. simium isolates infecting humans 51 and two isolates from brown howler monkeys revealed that P. simium is monophyletic within 52 the broader diversity of South American Plasmodium vivax, consistent with the hypothesis 53 that P. simium first infected non-human primates as a result of a host-switch from humans 54 carrying P. vivax. We provide molecular evidence that the current zoonotic infections of 55 people have likely resulted from multiple independent host switches, each seeded from a 56 different monkey infection. Very low levels of genetic diversity within P. simium genomes 57 and the absence of P. simium-P. vivax hybrids suggest that the P. simium population emerged 58 recently and has subsequently experienced a period of independent evolution in Platyrrhini 59 monkeys. We further find that Plasmodium Interspersed Repeat (PIR) genes, Plasmodium 60Helical Interspersed Subtelomeric (PHIST) genes and Tryptophan-Rich Antigens (TRAg) 61 genes in P. siumium are genetically divergent from P. vivax and are enriched for non-62 synonymous single nucleotide polymorphisms, consistent with the rapid evolution of these 63 genes. Analysis of genes involved in erythrocyte invasion revealed several notable differences 64 between P. vivax and P. simium, including large deletions within the coding region of the 65 Duffy Binding Protein 1 (DBP1) and Reticulocyte Binding Protein 2a (RBP2a) genes in P. 66 simium. Genotyping of P. simium isolates from non-human primates (NHPs) and zoonotic 67 human infections showed that a precise deletion of 38 amino acids in DBP1 is exclusively 67 present in all human infecting isolates, whereas non-human primate infecting isolates were 68 polymorphic for the deletion. We speculate that these deletions in the parasite-encoded key 69 erythrocyte invasion ligands and the additional rapid genetic changes have facilitated zoonotic 70 transfer to humans. Non-human primate malaria parasites can be considered a reservoir of 71 potential infectious human parasites that must be considered in any attempt of malaria 72 elimination. The genome of P. simium will thus form an important basis for future functional 73 characterizations on the mechanisms underlying malaria zoonosis. 74 75 76 Introduction 77 78 There are currently eight species of malaria parasites known to cause disease in humans; 79 Plasmodium falciparum, Plasmodium vivax, Plasmodium malariae, Plasmodium ovale 80 curtisi, Plasmodium ovale wallikeri, Plasmodium knowlesi, Plasmodium cynomolgi and 81 Plasmodium simium. The latter three species are more commonly parasitic on non-human 82 primates and have only relatively recently been shown to infect humans 1-3 . ...

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Section: Introductionmentioning

confidence: 99%

The genome of the zoonotic malaria parasite Plasmodium simium reveals adaptations to host-switching

Mourier

Alvarenga

Kaushik

et al. 2019

Preprint

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“…Targeting the 18S rRNA gene, 29 samples were analysed, of which 16 were positive for P. vivax, corresponding to 14 pools of A. K. cruzii collected in tree canopies and two livers from monkey bodies. The proposed protocol to differentiate samples between P. vivax and P. simium (13) were successful for seven samples, six resulting from DNA pools of A. K. cruzii and one from a simian liver.…”

Section: Methodsmentioning

confidence: 99%

Detection ofPlasmodium vivaxin a liver sample of a howler-monkey: one evidence more in favour of the identity betweenPlasmodium simiumandP. vivax

Buery

Duarte

Alencar

et al. 2020

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Introduction: The residual malaria of Atlantic Forest systems in Brazil occurs as an endemic disease with low frequency of cases. The chronological and spatial distance among the cases indicate an absence of fitness to the classical malaria cycle. This peculiar condition raised the suspicion of a reservoir, possibly the non-human primates. Simian and human malaria occur at the same places in that region, and there is already evidence of molecular identity between the simian parasites, Plasmodium simium and Plasmodium brasilianum, and the human parasites, Plasmodium vivax and Plasmodium malariae, respectively. Two different SNPs identified in the COX1 region of the Plasmodium vivax/simium of the Atlantic Forest reinforced its characterization as a zoonotic parasite. This finding supported the development of a PCR-RFLP protocol to identify such polymorphisms, and to monitor zoonotic malaria transmission. Methods: In the present work, we tested the above-mentioned PCR-RFLP protocol in unprecedented mosquitoes and simian samples collected in Espirito Santo State, Brazil (ES). Results: The parasite found in the simian sample was P. vivax, contrary to what the protocol should indicate. In the mosquito samples, the protocol disclosed both forms of the parasite. Conclusion: This result suggests that the previously published pair of SNPs, and, consequently, the PCR-RFLP protocol, are not able to distinguish the dynamics of Plasmodium spp. circulation in the Atlantic Forest endemic area of ES.

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“…All DNA samples were tested in triplicate for 18s rRNA Plasmodium genus-specifc gene [29,30], and then for cysteine proteinase P. vivax and ssrRNA P. malariae and P. falciparum genes, as previously described [30,31]. P. vivax- positive samples were submitted to P. simium differential diagnosis based on a mitochondrial SNP [2,10]. The molecular diagnosis was performed by Nested -PCR of coxI gene fragment and subsequent enzymatic digestion, using primers and protocol previously described [10].…”

Section: Methodsmentioning

confidence: 99%

“…P. vivax- positive samples were submitted to P. simium differential diagnosis based on a mitochondrial SNP [2,10]. The molecular diagnosis was performed by Nested -PCR of coxI gene fragment and subsequent enzymatic digestion, using primers and protocol previously described [10]. All the PCR products were visualized under UV light after electrophoresis on 2% agarose gels.…”

Section: Methodsmentioning

confidence: 99%

“…Two species of Plasmodium have been described in neotropical non-human primates (NHP): P. brasilianum Gonder e Berenberg-Gossler (1908) and P. simium , almost indistinguishable from the human malaria parasites P. malariae and P. vivax , respectively [5,6,9]. Besides subtle morphological variations [2,5], molecular markers such as microsatellites and single nucleotide polymorphisms (SNPs) were the only differences so far described between P. malariae and P. brasilianum and P. vivax and P. simium [2,10,11]. P. brasilianum has a broader distribution as has been found from México to Southern Brazil, infecting at least 11 genera of the five families of neotropical primates (Aotidae, Atelidae, Callitrichidae, Cebidae and Phiteciidae) [5,12–15].…”

Section: Introductionmentioning

confidence: 99%

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Howler monkeys are the reservoir of malaria parasites causing zoonotic infections in the Atlantic forest of Rio de Janeiro

Abreu¹,

Santos

Mello

et al. 2019

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BackgroundAlthough malaria transmission was eradicated from southeast Brazil, a significant increase in the number of Plasmodium vivax-like autochthonous human cases has been reported in remote areas of the Atlantic Forest in the last decades in Rio de Janeiro (RJ) state, including an outbreak in 2015-2016. The singular clinical and epidemiological aspects of several human cases combined with molecular and genetic data revealed that they were due to the non-human primate (NHP) parasite P. simium. The full understanding of the epidemiology of the autochthonous malaria in southeastern Brazil depends, however, upon the knowledge on the circulation of NHP Plasmodium in the foci and the determination of its reservoirs.MethodologyA large sampling effort was carried out in the Atlantic forest of RJ and its bordering states (Minas Gerais, São Paulo, Espírito Santo) for capture and examination of free-living NHPs. Blood and/or viscera were analyzed for Plasmodia infections through molecular and microscopic techniques.Principal findingsIn total, 146 NHPs of six species, from 30 counties in four states were tested. Howler monkeys (A. guariba clamitans) were the only NHP species found infected. In RJ, 26% of howlers were positive, among them 17% were found to be infected with P. simium. Importantly, specific single nucleotide polymorphisms were detected in all P. simium infected howlers regardless geographical origin of malaria foci. Interestingly, 71% of P. simium infected NHP were from the coastal slope of a mountain chain (Serra do Mar), where most human cases have been occurring. P. brasilianum/malariae was detected for the first time in 14% free-living howlers in RJ as well as in 25% of those from the Espírito Santo state. Moreover, malarial pigment was detected in spleen fragments of 50% of a subsample composed of howler monkeys found dead in both RJ and ES. All NHPs were negative for P. falciparum.Conclusions/SignificanceOur data indicate the howler monkeys as the main reservoir of the Atlantic forest human malaria in RJ and other sites in Southeast Brazil and reinforce its zoonotic nature.Author summaryThe present work consists of an unprecedented capture effort and large-scale field survey of plasmodial species in Non-human primates (NHPs) in RJ, a state recording a three-decade history of autochthonous human cases of benign tertian malaria pending epidemiological clarification of their origin. For the first time, we describe infection rates by Plasmodium sp.in free-living NHP, match the spatial distribution of P. simium in NHP with that of local human cases of benign tertian malaria due to this parasite, disclose howler monkeys as the only confirmed reservoir of this zoonotic malaria in the state and showed that specific single nucleotide polymorphisms were present in all P. simium infected howlers, regardless of the geographical origin of malaria foci. It is also the first time that P. brasilianum/malariae is recorded in free-living NHPs from Rio de Janeiro and the widespread distribution of this quartan-malaria parasite of zoonotic potential in the state is illustrated. Together, these findings increase the understanding about the simian malaria parasites in Atlantic Forests, as well as on the zoonotic character of autochthonous human malaria in Rio de Janeiro, providing subsidies for shaping surveillance and control.

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An assay for the identification of Plasmodium simium infection for diagnosis of zoonotic malaria in the Brazilian Atlantic Forest

Cited by 32 publications

References 33 publications

The genome of the zoonotic malaria parasite Plasmodium simium reveals adaptations to host-switching

The genome of the zoonotic malaria parasite Plasmodium simium reveals adaptations to host-switching

Detection ofPlasmodium vivaxin a liver sample of a howler-monkey: one evidence more in favour of the identity betweenPlasmodium simiumandP. vivax

Howler monkeys are the reservoir of malaria parasites causing zoonotic infections in the Atlantic forest of Rio de Janeiro

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