Your browser doesn't support javascript.
loading
Show: 20 | 50 | 100
Results 1 - 15 de 15
Filter
Add more filters










Publication year range
1.
Insect Sci ; 2024 Jan 21.
Article in English | MEDLINE | ID: mdl-38246860

ABSTRACT

In arthropods, hematophagy has arisen several times throughout evolution. This specialized feeding behavior offered a highly nutritious diet obtained during blood feeds. On the other hand, blood-sucking arthropods must overcome problems brought on by blood intake and digestion. Host blood complement acts on the bite site and is still active after ingestion, so complement activation is a potential threat to the host's skin feeding environment and to the arthropod gut enterocytes. During evolution, blood-sucking arthropods have selected, either in their saliva or gut, anticomplement molecules that inactivate host blood complement. This review presents an overview of the complement system and discusses the arthropod's salivary and gut anticomplement molecules studied to date, exploring their mechanism of action and other aspects related to the arthropod-host-pathogen interface. The possible therapeutic applications of arthropod's anticomplement molecules are also discussed.

2.
Rev Soc Bras Med Trop ; 53: e20200177, 2020.
Article in English | MEDLINE | ID: mdl-32935785

ABSTRACT

INTRODUCTION: Triatomine bugs are hematophagous insects that are extremely important in public health because they are natural vectors of Trypanosoma cruzi, the causative agent of Chagas disease. In this study, we aimed to assess the occurrence of triatomine species and the natural T. cruzi infection in the Guaribas Valley territory, an endemic region for Chagas disease in northeastern Brazil. METHODS: Insects were actively captured from July 2017 to October 2019 in the intra- and peridomiciles of 16 municipalities of the Guaribas Valley territory, in the southeast area of Piauí state. Triatomine species were identified following a taxonomic key, and natural infection was investigated through insects' fresh feces exams. RESULTS: A total of 430 triatomines were collected, including 211 nymphs and 219 adults. Of all collected specimens, 39 (9.1%) were from the intradomiciles and 391 (90.9%) from peridomiciles. Nine species, including two subspecies, could be identified: Triatoma brasiliensis brasiliensis, T. brasiliensis macromelasoma, T. pseudomaculata, T. sordida, T. juazeirensis, T. melanocephala, Panstrongylus lutzi, Rhodnius domesticus, R. nasutus, and R. robustus. T. brasiliensis were the most frequently collected bugs, representing 72% of all the identified insects. None of the examined invertebrates presented flagellate forms of T. cruzi. CONCLUSIONS: This is the first report of T. b. macromelasoma and T. juazeirensis in the Guaribas Valley territory. The persistence of triatomine species in the domiciles in an endemic area for Chagas disease emphasizes the relevance of entomological surveillance and vector control measures in the studied area.


Subject(s)
Chagas Disease , Triatoma , Triatominae , Trypanosoma cruzi , Animals , Brazil , Insect Vectors
3.
Rev. Soc. Bras. Med. Trop ; 53: e20200177, 2020. tab, graf
Article in English | Sec. Est. Saúde SP, Coleciona SUS, LILACS | ID: biblio-1136813

ABSTRACT

Abstract INTRODUCTION Triatomine bugs are hematophagous insects that are extremely important in public health because they are natural vectors of Trypanosoma cruzi, the causative agent of Chagas disease. In this study, we aimed to assess the occurrence of triatomine species and the natural T. cruzi infection in the Guaribas Valley territory, an endemic region for Chagas disease in northeastern Brazil. METHODS Insects were actively captured from July 2017 to October 2019 in the intra- and peridomiciles of 16 municipalities of the Guaribas Valley territory, in the southeast area of Piauí state. Triatomine species were identified following a taxonomic key, and natural infection was investigated through insects' fresh feces exams. RESULTS A total of 430 triatomines were collected, including 211 nymphs and 219 adults. Of all collected specimens, 39 (9.1%) were from the intradomiciles and 391 (90.9%) from peridomiciles. Nine species, including two subspecies, could be identified: Triatoma brasiliensis brasiliensis, T. brasiliensis macromelasoma, T. pseudomaculata, T. sordida, T. juazeirensis, T. melanocephala, Panstrongylus lutzi, Rhodnius domesticus, R. nasutus, and R. robustus. T. brasiliensis were the most frequently collected bugs, representing 72% of all the identified insects. None of the examined invertebrates presented flagellate forms of T. cruzi. CONCLUSIONS: This is the first report of T. b. macromelasoma and T. juazeirensis in the Guaribas Valley territory. The persistence of triatomine species in the domiciles in an endemic area for Chagas disease emphasizes the relevance of entomological surveillance and vector control measures in the studied area.


Subject(s)
Animals , Triatoma , Trypanosoma cruzi , Triatominae , Chagas Disease , Brazil , Insect Vectors
4.
Rev Inst Med Trop Sao Paulo ; 61: e38, 2019 Aug 08.
Article in English | MEDLINE | ID: mdl-31411268

ABSTRACT

The triatomine insect Panstrongylus megistus , one of the most important Chagas disease vectors in Brazil, presents salivary molecules pharmacologically active to counteract homeostatic responses from the host, including inhibitors of the human complement system, a major effector of immune responses. The aim of the present study was to investigate the effect of P. megistus salivary gland extract (SGE) on the complement system from different host species and characterize the inhibitory effect of SGE and intestinal contents on human complement. Glands and midguts from fourth instar nymphs were used. Hemolytic assays were performed with sheep erythrocytes as complement activators by using human, rats and chickens sera in the presence or absence of SGE. An ELISA assay was carried out detect deposition of the C3b component on IgG- or agarose-sensitized microplates, in the presence or absence of SGE or midgut contents. P. megistus SGE was able to significantly inhibit the complement of the three studied species (human, rat and chiken). Both, SGE and midgut contents inhibited C3b deposition in either the classical or the alternative pathways. As conclusions, SGE and midgut from P. megistus possess anti-complement activity. The inhibitors are effective against different host species and act on the initial steps of the complement system cascade. These inhibitors may have a role in blood feeding and Trypanosoma cruzi transmission by the vector.


Subject(s)
Complement System Proteins/drug effects , Digestive System/chemistry , Insect Vectors , Panstrongylus , Salivary Glands/chemistry , Animals , Chagas Disease/transmission , Chickens , Humans , Rats , Sheep
5.
Sci Rep ; 9(1): 6764, 2019 05 01.
Article in English | MEDLINE | ID: mdl-31043627

ABSTRACT

Ornithodoros rostratus is a South American argasid tick which importance relies on its itchy bite and potential as disease vector. They feed on a wide variety of hosts and secrete different molecules in their saliva and intestinal content that counteract host defences and help to accommodate and metabolize the relatively large quantity of blood upon feeding. The present work describes the transcriptome profile of salivary gland (SG) and midgut (MG) of O. rostratus using Illumina sequencing. A total of 8,031 contigs were assembled and assigned to different functional classes. Secreted proteins were the most abundant in the SG and accounted for ~67% of all expressed transcripts with contigs with identity to lipocalins and acid tail proteins being the most representative. On the other hand, immunity genes were upregulated in MG with a predominance of defensins and lysozymes. Only 10 transcripts in SG and 8 in MG represented ~30% of all RNA expressed in each tissue and one single contig (the acid tail protein ORN-9707) represented ~7% of all expressed contigs in SG. Results highlight the functional difference of each organ and identified the most expressed classes and contigs of O. rostratus SG and MG.


Subject(s)
Arthropod Proteins/metabolism , Ornithodoros/metabolism , Proteome/analysis , RNA-Seq/methods , Salivary Glands/metabolism , Salivary Proteins and Peptides/metabolism , Transcriptome , Animals , Arthropod Proteins/genetics , Computational Biology , Evolution, Molecular , Ornithodoros/genetics , Ornithodoros/growth & development , Phylogeny , Salivary Proteins and Peptides/genetics
6.
Insect Biochem Mol Biol ; 92: 12-20, 2018 01.
Article in English | MEDLINE | ID: mdl-29128668

ABSTRACT

Anopheline mosquitoes are vectors of malaria parasites. Their saliva contains anti-hemostatic and immune-modulator molecules that favor blood feeding and parasite transmission. In this study, we describe the inhibition of the alternative pathway of the complement system (AP) by Anopheles aquasalis salivary gland extracts (SGE). According to our results, the inhibitor present in SGE acts on the initial step of the AP blocking deposition of C3b on the activation surfaces. Properdin, which is a positive regulatory molecule of the AP, binds to SGE. When SGE was treated with an excess of properdin, it was unable to inhibit the AP. Through SDS-PAGE analysis, A. aquasalis presented a salivary protein with the same molecular weight as recombinant complement inhibitors belonging to the SG7 family described in the saliva of other anopheline species. At least some SG7 proteins bind to properdin and are AP inhibitors. Searching for SG7 proteins in the A. aquasalis genome, we retrieved a salivary protein that shared an 85% identity with albicin, which is the salivary alternative pathway inhibitor from A. albimanus. This A. aquasalis sequence was also very similar (81% ID) to the SG7 protein from A. darlingi, which is also an AP inhibitor. Our results suggest that the salivary complement inhibitor from A. aquasalis is an SG7 protein that can inhibit the AP by binding to properdin and abrogating its stabilizing activity. Albicin, which is the SG7 from A. albimanus, can directly inhibit AP convertase. Given the high similarity of SG7 proteins, the SG7 from A. aquasalis may also directly inhibit AP convertase in the absence of properdin.


Subject(s)
Anopheles/genetics , Insect Proteins/genetics , Properdin/genetics , Salivary Proteins and Peptides/genetics , Amino Acid Sequence , Animals , Anopheles/metabolism , Insect Proteins/chemistry , Insect Proteins/metabolism , Phylogeny , Properdin/chemistry , Properdin/metabolism , Saliva/chemistry , Salivary Proteins and Peptides/chemistry , Salivary Proteins and Peptides/metabolism
7.
Front Immunol ; 8: 1065, 2017.
Article in English | MEDLINE | ID: mdl-28912782

ABSTRACT

Saliva of the blood feeding sand fly Lutzomyia longipalpis was previously shown to inhibit the alternative pathway (AP) of the complement system. Here, we have identified Lufaxin, a protein component in saliva, as the inhibitor of the AP. Lufaxin inhibited the deposition of C3b, Bb, Properdin, C5b, and C9b on agarose-coated plates in a dose-dependent manner. It also inhibited the activation of factor B in normal serum, but had no effect on the components of the membrane attack complex. Surface plasmon resonance (SPR) experiments demonstrated that Lufaxin stabilizes the C3b-B proconvertase complex when passed over a C3b surface in combination with factor B. Lufaxin was also shown to inhibit the activation of factor B by factor D in a reconstituted C3b-B, but did not inhibit the activation of C3 by reconstituted C3b-Bb. Proconvertase stabilization does not require the presence of divalent cations, but addition of Ni2+ increases the stability of complexes formed on SPR surfaces. Stabilization of the C3b-B complex to prevent C3 convertase formation (C3b-Bb formation) is a novel mechanism that differs from previously described strategies used by other organisms to inhibit the AP of the host complement system.

8.
PLoS Negl Trop Dis ; 11(3): e0005374, 2017 03.
Article in English | MEDLINE | ID: mdl-28278244

ABSTRACT

BACKGROUND: Immunity to the sand fly salivary protein SALO (Salivary Anticomplement of Lutzomyia longipalpis) protected hamsters against Leishmania infantum and L. braziliensis infection and, more recently, a vaccine combination of a genetically modified Leishmania with SALO conferred strong protection against L. donovani infection. Because of the importance of SALO as a potential component of a leishmaniasis vaccine, a plan to produce this recombinant protein for future scale manufacturing as well as knowledge of its structural characteristics are needed to move SALO forward for the clinical path. METHODOLOGY/PRINCIPAL FINDINGS: Recombinant SALO was expressed as a soluble secreted protein using Pichia pastoris, rSALO(P), with yields of 1g/L and >99% purity as assessed by SEC-MALS and SDS-PAGE. Unlike its native counterpart, rSALO(P) does not inhibit the classical pathway of complement; however, antibodies to rSALO(P) inhibit the anti-complement activity of sand fly salivary gland homogenate. Immunization with rSALO(P) produces a delayed type hypersensitivity response in C57BL/6 mice, suggesting rSALO(P) lacked anti-complement activity but retained its immunogenicity. The structure of rSALO(P) was solved by S-SAD at Cu-Kalpha to 1.94 Å and refined to Rfactor 17%. SALO is ~80% helical, has no appreciable structural similarities to any human protein, and has limited structural similarity in the C-terminus to members of insect odorant binding proteins. SALO has three predicted human CD4+ T cell epitopes on surface exposed helices. CONCLUSIONS/SIGNIFICANCE: The results indicate that SALO as expressed and purified from P. pastoris is suitable for further scale-up, manufacturing, and testing. SALO has a novel structure, is not similar to any human proteins, is immunogenic in rodents, and does not have the anti-complement activity observed in the native salivary protein which are all important attributes to move this vaccine candidate forward to the clinical path.


Subject(s)
Psychodidae/chemistry , Recombinant Proteins/immunology , Salivary Proteins and Peptides/immunology , Animals , Gene Expression , Mice, Inbred C57BL , Pichia/genetics , Pichia/metabolism , Protein Conformation , Recombinant Proteins/administration & dosage , Recombinant Proteins/chemistry , Recombinant Proteins/genetics , Salivary Proteins and Peptides/administration & dosage , Salivary Proteins and Peptides/chemistry , Salivary Proteins and Peptides/genetics
9.
Mem Inst Oswaldo Cruz ; 112(1): 53-63, 2017 Jan 01.
Article in English | MEDLINE | ID: mdl-28076469

ABSTRACT

Canine visceral leishmaniasis (CVL) diagnosis is still a challenge in endemic areas with limited diagnostic resources. This study proposes a score with the potential to distinguish positive CVL cases from negative ones. We studied 265 dogs that tested positive for CVL on ELISA and parasitological tests. A score ranging between 0 and 19 was recorded on the basis of clinical signs. Dogs with CVL had an overall higher positivity of the majority of clinical signs than did dogs without CVL or with ehrlichiosis. Clinical signs such as enlarged lymph nodes (83.93%), muzzle/ear lesions (55.36%), nutritional status (51.79%), bristle condition (57.14%), pale mucosal colour (48.21%), onychogryphosis (58.93%), skin lesion (39.28%), bleeding (12.50%), muzzle depigmentation (41.07%), alopecia (39.29%), blepharitis (21.43%), and keratoconjunctivitis (42.86%) were more frequent in dogs with CVL than in dogs with ehrlichiosis or without CVL. Moreover, the clinical score increased according to the positivity of all diagnostic tests (ELISA, p < 0.001; parasite culture, p = 0.0021; and smear, p = 0.0003). Onychogryphosis (long nails) [odds ratio (OR): 3.529; 95% confidence interval (CI): 1.832-6.796; p < 0.001], muzzle depigmentation (OR: 4.651; 95% CI: 2.218-9.750; p < 0.001), and keratoconjunctivitis (OR: 5.400; 95% CI: 2.549-11.441; p < 0.001) were highly associated with CVL. Interestingly, a score cut-off value ≥ 6 had an area under the curve of 0.717 (p < 0.0001), sensitivity of 60.71%, and specificity of 73.64% for CVL diagnosis. The clinical sign-based score for CVL diagnosis suggested herein can help veterinarians reliably identify dogs with CVL in endemic areas with limited diagnostic resources.


Subject(s)
Antibodies, Protozoan/blood , Dog Diseases/diagnosis , Leishmania infantum/immunology , Leishmaniasis, Visceral/veterinary , Animals , Brazil/epidemiology , Dog Diseases/epidemiology , Dogs , Enzyme-Linked Immunosorbent Assay/veterinary , Female , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/epidemiology , Male , Sensitivity and Specificity
10.
Mem. Inst. Oswaldo Cruz ; 112(1): 53-63, Jan. 2017. tab, graf
Article in English | LILACS | ID: biblio-841749

ABSTRACT

Canine visceral leishmaniasis (CVL) diagnosis is still a challenge in endemic areas with limited diagnostic resources. This study proposes a score with the potential to distinguish positive CVL cases from negative ones. We studied 265 dogs that tested positive for CVL on ELISA and parasitological tests. A score ranging between 0 and 19 was recorded on the basis of clinical signs. Dogs with CVL had an overall higher positivity of the majority of clinical signs than did dogs without CVL or with ehrlichiosis. Clinical signs such as enlarged lymph nodes (83.93%), muzzle/ear lesions (55.36%), nutritional status (51.79%), bristle condition (57.14%), pale mucosal colour (48.21%), onychogryphosis (58.93%), skin lesion (39.28%), bleeding (12.50%), muzzle depigmentation (41.07%), alopecia (39.29%), blepharitis (21.43%), and keratoconjunctivitis (42.86%) were more frequent in dogs with CVL than in dogs with ehrlichiosis or without CVL. Moreover, the clinical score increased according to the positivity of all diagnostic tests (ELISA, p < 0.001; parasite culture, p = 0.0021; and smear, p = 0.0003). Onychogryphosis (long nails) [odds ratio (OR): 3.529; 95% confidence interval (CI): 1.832-6.796; p < 0.001], muzzle depigmentation (OR: 4.651; 95% CI: 2.218-9.750; p < 0.001), and keratoconjunctivitis (OR: 5.400; 95% CI: 2.549-11.441; p < 0.001) were highly associated with CVL. Interestingly, a score cut-off value ≥ 6 had an area under the curve of 0.717 (p < 0.0001), sensitivity of 60.71%, and specificity of 73.64% for CVL diagnosis. The clinical sign-based score for CVL diagnosis suggested herein can help veterinarians reliably identify dogs with CVL in endemic areas with limited diagnostic resources.


Subject(s)
Animals , Male , Female , Dogs , Leishmania infantum/immunology , Dog Diseases/diagnosis , Dog Diseases/epidemiology , Leishmaniasis, Visceral/diagnosis , Leishmaniasis, Visceral/veterinary , Leishmaniasis, Visceral/epidemiology , Antigens, Protozoan/blood , Brazil/epidemiology , Enzyme-Linked Immunosorbent Assay/veterinary , Sensitivity and Specificity , Antibodies
12.
J Immunol ; 197(2): 599-610, 2016 07 15.
Article in English | MEDLINE | ID: mdl-27307559

ABSTRACT

The complement system present in circulating blood is an effective mechanism of host defense, responsible for the killing of pathogens and the production of potent anaphylatoxins. Inhibitors of the complement system have been described in the saliva of hematophagous arthropods that are involved in the protection of digestive tissues against complement system-mediated damage. In this study, we describe albicin, a novel inhibitor of the alternative pathway of complement from the salivary glands of the malaria vector, Anopheles albimanus The inhibitor was purified from salivary gland homogenates by reverse-phase HPLC and identified by mass spectrometry as a small (13.4-kDa) protein related to the gSG7 protein of Anopheles gambiae and Anopheles stephensi Recombinant albicin was produced in Escherichia coli and found to potently inhibit lysis of rabbit erythrocytes in assays of the alternative pathway while having no inhibitory effect on the classical or lectin pathways. Albicin also inhibited the deposition of complement components on agarose-coated plates, although it could not remove previously bound components. Antisera produced against recombinant albicin recognized both the native and recombinant inhibitors and also blocked their activities in in vitro assays. Using surface plasmon resonance and enzymatic assays, we found that albicin binds and stabilizes the C3-convertase complex (C3bBb) formed on a properdin surface and inhibits the convertase activity of a reconstituted C3bBb complex in solution. The data indicate that albicin specifically recognizes the activated form of the complex, allowing more efficient inhibition by an inhibitor whose quantity is limited.


Subject(s)
Anopheles/immunology , Complement Pathway, Alternative/immunology , Insect Proteins/immunology , Saliva/immunology , Salivary Proteins and Peptides/immunology , Animals , Blotting, Western , Chromatography, High Pressure Liquid , Enzyme-Linked Immunosorbent Assay , Female , Insect Proteins/genetics , Insect Proteins/metabolism , Mass Spectrometry , Polymerase Chain Reaction , Rabbits , Surface Plasmon Resonance
13.
Sci Rep ; 6: 19300, 2016 Jan 13.
Article in English | MEDLINE | ID: mdl-26758086

ABSTRACT

Blood-feeding insects inject potent salivary components including complement inhibitors into their host's skin to acquire a blood meal. Sand fly saliva was shown to inhibit the classical pathway of complement; however, the molecular identity of the inhibitor remains unknown. Here, we identified SALO as the classical pathway complement inhibitor. SALO, an 11 kDa protein, has no homology to proteins of any other organism apart from New World sand flies. rSALO anti-complement activity has the same chromatographic properties as the Lu. longipalpis salivary gland homogenate (SGH)counterparts and anti-rSALO antibodies blocked the classical pathway complement activity of rSALO and SGH. Both rSALO and SGH inhibited C4b deposition and cleavage of C4. rSALO, however, did not inhibit the protease activity of C1s nor the enzymatic activity of factor Xa, uPA, thrombin, kallikrein, trypsin and plasmin. Importantly, rSALO did not inhibit the alternative or the lectin pathway of complement. In conclusion our data shows that SALO is a specific classical pathway complement inhibitor present in the saliva of Lu. longipalpis. Importantly, due to its small size and specificity, SALO may offer a therapeutic alternative for complement classical pathway-mediated pathogenic effects in human diseases.


Subject(s)
Complement Inactivating Agents/pharmacology , Complement Pathway, Classical/drug effects , Insect Proteins/pharmacology , Psychodidae/immunology , Psychodidae/metabolism , Saliva/metabolism , Animals , Chromatography, High Pressure Liquid , Complement Activation/drug effects , Complement C1/antagonists & inhibitors , Complement C1/immunology , Complement C1/metabolism , Complement C4/antagonists & inhibitors , Complement C4/immunology , Complement C4/metabolism , Humans , Recombinant Proteins/pharmacology
14.
PLoS One ; 8(11): e79787, 2013.
Article in English | MEDLINE | ID: mdl-24255715

ABSTRACT

BACKGROUND: Lutzomyia longipalpis is the vector of Leishmania infantum in the New World, and its saliva inhibits classical and alternative human complement system pathways. This inhibition is important in protecting the insect´s midgut from damage by the complement. L. longipalpis is a promiscuous blood feeder and must be protected against its host's complement. The objective of this study was to investigate the action of salivary complement inhibitors on the sera of different host species, such as dogs, guinea pigs, rats and chickens, at a pH of 7.4 (normal blood pH) and 8.15 (the midgut pH immediately after a blood meal). We also investigated the role of the chicken complement system in Leishmania clearance in the presence and absence of vector saliva. RESULTS: The saliva was capable of inhibiting classical pathways in dogs, guinea pigs and rats at both pHs. The alternative pathway was not inhibited except in dogs at a pH of 8.15. The chicken classical pathway was inhibited only by high concentrations of saliva and it was better inhibited by the midgut contents of sand flies. Neither the saliva nor the midgut contents had any effect on the avian alternative pathway. Fowl sera killed L. infantum promastigotes, even at a low concentration (2%), and the addition of L. longipalpis saliva did not protect the parasites. The high body temperature of chickens (40°C) had no effect on Leishmania viability during our assays. CONCLUSION: Salivary inhibitors act in a species-specific manner. It is important to determine their effects in the natural hosts of Leishmania infantum because they act on canid and rodent complements but not on chickens (which do not harbour the parasite). Moreover, we concluded that the avian complement system is the probable mechanism through which chickens eliminate Leishmania and that their high body temperature does not influence this parasite.


Subject(s)
Complement System Proteins/immunology , Leishmania infantum/immunology , Psychodidae/immunology , Psychodidae/parasitology , Saliva/immunology , Animals , Chickens , Complement Activation , Complement C3b/immunology , Complement Pathway, Alternative , Complement Pathway, Classical , Cytotoxicity, Immunologic , Dogs , Female , Guinea Pigs , Host-Pathogen Interactions/immunology , Hydrogen-Ion Concentration , Kinetics , Rats
15.
Acta Trop ; 107(3): 230-7, 2008 Sep.
Article in English | MEDLINE | ID: mdl-18606150

ABSTRACT

The phlebotomine sand fly Lutzomyia longipalpis takes blood from a variety of wild and domestic animals and transmits Leishmania (Leishmania) infantum chagasi, etiological agent of American visceral leishmaniasis. Blood meal identification in sand flies has depended largely on serological methods but a new protocol described here uses filter-based technology to stabilise and store blood meal DNA, allowing subsequent PCR identification of blood meal sources, as well as parasite detection, in blood-fed sand flies. This technique revealed that 53.6% of field-collected sand flies captured in the back yards of houses in Teresina (Brazil) had fed on chickens. The potential applications of this technique in epidemiological studies and strategic planning for leishmaniasis control programmes are discussed.


Subject(s)
Blood , Feeding Behavior , Parasitology/methods , Psychodidae , Specimen Handling/methods , Animals , Animals, Domestic , Animals, Wild , Brazil , Cricetinae , Female , Humans , Leishmaniasis, Visceral/epidemiology , Leishmaniasis, Visceral/prevention & control , Polymerase Chain Reaction/methods
SELECTION OF CITATIONS
SEARCH DETAIL
...