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1.
Mem. Inst. Oswaldo Cruz ; 115: e200157, 2020. tab, graf
Article in English | LILACS, SES-SP | ID: biblio-1135268

ABSTRACT

BACKGROUND In Acre state, Brazil, the dissemination of cutaneous leishmaniasis has increased in recent years, with limited knowledge of the potential Leishmania spp. vectors involved. OBJECTIVES Here, data concerning the sandfly fauna of Brasiléia municipality, Leishmania DNA-detection rates and the identification of blood meal sources of insects captured in 2013-2015 are presented. METHODS Parasite detection in female sandflies was performed individually by multiplex polymerase chain reaction (PCR) (Leishmania kDNA/sandfly cacophony-gene), with the identification of Leishmania spp. by hsp70-PCR and sequencing. The identification of blood gut-content from fed females was performed by cyt b-PCR and sequencing. FINDINGS A total of 4,473 sandflies were captured. A subgroup of 864 non-blood-fed females evaluated for the presence of Leishmania DNA showed 2.9% positivity for Leishmania (Viannia) braziliensis and L. (V.) guyanensis. The identification of blood meal sources was performed in 96 blood-fed females, allowing the identification of 13 vertebrate species. In nine/96 fed females, DNA from L. (V.) shawi, L. (V.) guyanensis, L. (V.) braziliensis and Endotrypanum sp. was detected. MAIN CONCLUSIONS In Brumptomyia sp. and Evandromyia termitophila, the first report of Leishmania DNA-detection is provided in Acre; Nyssomyia shawi is implicated as potential vector of L. (V.) braziliensis and L. (V.) guyanensis for the first time in Brazil.


Subject(s)
Animals , Female , Psychodidae/parasitology , DNA/analysis , Insect Vectors/genetics , Leishmania/genetics , Psychodidae/classification , Brazil , Polymerase Chain Reaction , DNA, Protozoan/analysis , Leishmaniasis, Cutaneous/transmission , Insect Vectors/classification , Insect Vectors/parasitology , Leishmania/isolation & purification
2.
Rev. bras. parasitol. vet ; 28(1): 1-11, Jan.-Mar. 2019. tab
Article in English | LILACS | ID: biblio-990813

ABSTRACT

Abstract Tritrichomonas foetus is a parasite that has been definitively identified as an agent of trichomonosis, a disease characterized by chronic diarrhea. T. foetus colonizes portions of the feline large intestine, and manifests as chronic and recurrent diarrhea with mucus and fresh blood, which is often unresponsive to common drugs. Diagnosis of a trichomonad infection is made by either the demonstration of the trophozoite on a direct fecal smear, fecal culture and subsequent microscopic examination of the parasite, or extraction of DNA in feces and amplification by the use of molecular tools. T. foetus is commonly misidentified as other flagellate protozoa such as Giardia duodenalis and Pentatrichomonas hominis. Without proper treatment, the diarrhea may resolve spontaneously in months to years, but cats can remain carriers of the parasite. This paper intends to serve as a source of information for investigators and veterinarians, reviewing the most important aspects of feline trichomonosis, such as trichomonad history, biology, clinical manifestations, pathogenesis, world distribution, risk factors, diagnosis, and treatment.


Resumo Tritrichomonas foetus é um parasito que foi identificado definitivamente como agente de tricomoníase, caracterizada por diarreia crônica. T. foetus coloniza porções do intestino grosso dos felinos e se manifesta como uma diarreia crônica e recorrente, com muco e sangue, geralmente irresponsiva às drogas comumente usadas no tratamento. O diagnóstico da infecção por tricomonadídeos é feito pela demonstração de trofozoítos no exame direto de fezes frescas, cultura fecal e subsequente exame microscópico ou extração do DNA do parasito na amostra fecal e amplificação, utilizando-se técnicas moleculares. T. foetus é comumente confundido com outros protozoários flagelados, como Giardia duodenalis e Pentatrichomonas hominis. Sem tratamento adequado, a diarreia pode cessar espontaneamente em meses ou anos, porém os gatos podem permanecer portadores do parasito. Esse artigo pretende servir como fonte de informação para pesquisadores e veterinários, revisando os mais importantes aspectos da tricomoníase felina, como histórico, biologia, manifestações clínicas, patogênese, distribuição mundial, fatores de risco, diagnóstico e tratamento.


Subject(s)
Animals , Cats , Protozoan Infections, Animal/diagnosis , Protozoan Infections, Animal/drug therapy , Protozoan Infections, Animal/epidemiology , Tritrichomonas foetus/isolation & purification , Diarrhea/veterinary , Feces/parasitology , Cat Diseases/diagnosis , Cat Diseases/drug therapy , Cat Diseases/epidemiology , Polymerase Chain Reaction , Risk Factors , DNA, Protozoan/analysis , Tritrichomonas foetus/genetics , Diarrhea/parasitology
3.
Rev. Soc. Bras. Med. Trop ; 52: e20180537, 2019. tab, graf
Article in English | LILACS | ID: biblio-990433

ABSTRACT

Abstract INTRODUCTION: Elimination of malaria in areas of interrupted transmission warrants careful case assessment to avoid the reintroduction of this disease. Occasional malaria cases are reported among visitors of the Atlantic Forest area of Brazil, while data on residents of this area are scarce. METHODS: A sectional study was carried out to examine 324 individuals living in a municipality where autochthonous cases were detected. RESULTS: Asymptomatic Plasmodium infections were detected in 2.8% of the individuals by polymerase chain reaction (PCR), with one case of P. falciparum (0.3%), two cases of P. vivax (0.6%), and six cases of P. malariae (1.9%). The thick blood smears were negative in all individuals. Serological tests performed in 314 subjects were reactive in 11.1%, with 3.5% for P. falciparum and 7.7% for P. vivax. A subsample of 42 reactive individuals for any Plasmodium species showed P. malariae in 30.9% of specimens. Individuals who entered the Atlantic Forest region were 2.7 times more likely to exhibit reactive serology for P. vivax compared with individuals who did not enter this region (p<0.05). Children <15 years had a higher chance of reactive serology for P. falciparum and P. vivax than individuals ≥15 years of age (p<0.05). Individuals living in the Paraiso district had a higher chance of reactive serology for P. vivax compared to other districts (p<0.05). No associations were found between sex, past exposure to malaria, or serological response to antibodies of any Plasmodium species. CONCLUSIONS: The implications of these results for the elimination of malaria were discussed.


Subject(s)
Humans , Male , Female , Adult , Aged , Aged, 80 and over , Malaria, Vivax/diagnosis , Malaria, Vivax/transmission , Malaria, Falciparum/diagnosis , Malaria, Falciparum/transmission , Brazil/epidemiology , Enzyme-Linked Immunosorbent Assay , Polymerase Chain Reaction , Cross-Sectional Studies , DNA, Protozoan/analysis , Malaria, Vivax/epidemiology , Malaria, Falciparum/epidemiology , Asymptomatic Infections/epidemiology , Antigens, Protozoan/immunology
4.
Rev. bras. parasitol. vet ; 27(4): 481-487, Oct.-Dec. 2018. tab
Article in English | LILACS | ID: biblio-977925

ABSTRACT

Abstract Recent genetic population studies on Toxoplasma gondii in Brazil have shown large genetic variability. The objective of the present study was to isolate and genotypically characterize T. gondii from free-ranging and captive wild mammals and birds in Pernambuco state, Brazil. Fragments of heart, brain, skeletal muscle and diaphragm tissue from 71 birds and 34 mammals, which were either free-ranging or captive, were collected. Samples from 32 of these animals were subjected to bioassays in mice. Samples from the remaining 73 animals underwent biomolecular diagnosis, using PCR technique, targeting a repetitive DNA fragment of 529 bp in T. gondii. A non-virulent isolate (TgButstBrPE1) was obtained from a free-ranging striated heron (Butorides striata) and, based on primary samples, seven animals were found to be positive. The primary samples and the isolate obtained were subjected to PCR-RFLP using the markers SAG1, 5'3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico and CS3. ToxoDB-RFLP genotype #13 from the striated heron isolate and Type BrIII genotype from a captive otter ( Lontra longicaudis) (PS-TgLonloBrPE1) were obtained. The present study describes the first isolation and genotypic characterization of T. gondii in free-ranging striated heron, and the first genotypic characterization of T. gondii in a captive otter.


Resumo Recentes estudos genéticos nas populações deste parasita no Brasil têm mostrado grande variabilidade genética. O objetivo do presente estudo foi isolar e caracterizar genotipicamente T. gondii de aves e mamíferos de vida livre e de cativeiro no estado de Pernambuco, Brazil. Fragmentos de tecido do coração, cérebro, músculo esquelético e diafragma de 71 aves e 34 mamíferos de vida livre ou cativeiro foram colhidos. Amostras de 32 destes animais foram submetidas a bioensaios em camundongos. As amostras dos 73 animais restantes foram submetidas a diagnóstico biomolecular usando a técnica de PCR, tendo como alvo o fragmento repetitivo de 529 pb do DNA de T. gondii. Dentre os 32 bioensaios conduzidos, obteve-se um isolado não-virulento (TgButstBrPE1) de um socozinho (Butorides striata ) de vida livre, e dentre as amostras primárias, sete animais foram positivos. As amostras primárias e o isolado foram submetidos a PCR-RFLP usando os marcadores SAG1, 5'3'SAG2, alt.SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, L358, PK1, Apico e CS3. Foram obtidos o genótipo ToxoDB-RFLP #13 do isolado do socozinho e o genótipo Type BrIII de uma lontra (Lontra longicaudis) de cativeiro (PS-TgLonloBrPE1). O presente estudo descreve o primeiro isolamento e caracterização genotípica de T. gondii em socozinho de vida livre, e a primeira caracterização genotípica de T. gondii em lontra em cativeiro.


Subject(s)
Animals , Mice , Toxoplasma/isolation & purification , Birds/parasitology , Antibodies, Protozoan/blood , DNA, Protozoan/analysis , Mammals/parasitology , Toxoplasma/genetics , Toxoplasma/immunology , Genetic Variation , Polymorphism, Restriction Fragment Length , Brazil , Polymerase Chain Reaction , Genotype , Mammals/classification
5.
Rev. bras. parasitol. vet ; 27(3): 384-389, July-Sept. 2018. tab
Article in English | LILACS | ID: biblio-1042481

ABSTRACT

Abstract Toxoplasma gondii presents a high prevalence worldwide, infecting several animals. Felines are considered the definitive hosts and among the intermediate hosts we highlight mammals and birds. The man can become infected by ingesting tissue cysts present in birds and mammals. Biological and molecular aspects of T. gondii allows a better understanding of the epidemiology of toxoplasmosis. This work is a serologic screening of 58 chickens grown (Gallus gallus domesticus) for human consumption in Espírito Santo State, by means of indirect haemagglutination assay (IHA). Thirteen chickens tested positive for anti-T. gondii antibodies. The heart and brain of five positive chickens were harvested, treated with pepsin and inoculated separately, in two Swiss mice, intraperitoneally. Tachyzoites were observed in the peritoneum of all the animals, between seven and 10 days after the inoculum. Ten isolates were obtained and biologically characterised in BALB/c mice inoculated with 101 to 104 tachyzoites. All isolates were classified as virulent or intermediately virulent. Isolates were genotyped by means of polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, revealing three different genotypes. None of the isolates exhibited the clonal type I, II or III genotype. No genotypic differences were observed between the isolates from the brain or heart from the same bird.


Resumo Toxoplasma gondii apresenta alta prevalência mundial, capaz de infectar diversos animais. Felinos são considerados os hospedeiros definitivos e entre os hospedeiros intermediários destacamos os mamíferos e as aves. O homem pode se infectar ingerindo cistos teciduais presentes na carne das aves e mamíferos. O conhecimento dos aspectos biológicos e moleculares do parasito possibilitam melhor entendimento da epidemiologia da toxoplasmose. Neste trabalho foi realizada triagem sorológica por hemaglutinação indireta (HI) em 58 galinhas caipiras (Gallus gallus domesticus) utilizadas para consumo humano, provenientes do estado do Espírito Santo, Brasil. Treze galinhas apresentaram sorologia positiva para T. gondii. O coração e o cérebro de cinco galinhas positivas foram colhidos, tratados com pepsina e inoculados separadamente, em dois camundongos Swiss, por via intraperitoneal. Observou-se taquizoítos no peritônio de todos os camundongos, entre sete e 10 dias após o inóculo. Foram obtidos 10 novos isolados de T. gondii os quais foram estudados em camundongos BALB/C inoculados com 101 a 104 taquizoítos por animal. Todos os isolados foram considerados virulentos ou de virulência intermediária. A caracterização molecular dos isolados, realizada por PCR-RFLP, demonstrou a ocorrência de três genótipos distintos. Nenhum isolado apresentou genótipo clonal ou linhagem clonal do Brasil. Não foi observada diferença molecular (PCR-RFLP) entre os isolados obtidos a partir do cérebro ou do coração da mesma ave. Dois isolados já haviam sido relatados na literatura como causadores de doenças em humanos.


Subject(s)
Female , Mice , Poultry Diseases/parasitology , Toxoplasma/pathogenicity , Antibodies, Protozoan/blood , Chickens/parasitology , Toxoplasmosis, Animal/diagnosis , Poultry Diseases/diagnosis , Toxoplasma/isolation & purification , Toxoplasma/genetics , Polymorphism, Restriction Fragment Length , Brazil , Agglutination Tests , Polymerase Chain Reaction , DNA, Protozoan/analysis , Genotype , Mice, Inbred BALB C
6.
Rev. Soc. Bras. Med. Trop ; 51(4): 467-474, July-Aug. 2018. graf
Article in English | LILACS | ID: biblio-957441

ABSTRACT

Abstract INTRODUCTION: The detection of Trypanosoma cruzi in tissue samples is important in many situations, such as testing of the reactivation of the infection. The detection of T. cruzi nests in endomyocardial biopsies (EMB) may be useful to evaluate graft rejection. Given their scarcity, such nests are not routinely identified. To increase the diagnosis sensitivity, immunohistochemistry (IHC) may serve as a promising strategy. Here, we validate an antiserum for the detection of T. cruzi infection by IHC. METHODS: We used 1) positive controls (PCs) - 13 EMB, 12 skin biopsies, and 1 heart with T. cruzi nests as sections stained with hematoxylin and eosin (HE); 2) negative controls - a) 10 explant hearts and 10 EMB with no amastigote nests or clinical/laboratory signs of chagasic infection; and b) eight samples with leishmaniasis, toxoplasmosis, or histoplasmosis; and 3) Cases - 31 EMB of chagasic patients with no parasite nests in HE sections but detected positive for T. cruzi DNA by polymerase chain reaction. As a primary antibody, a hyperimmune serum from T. cruzi-infected rabbits was used. RESULTS: IHC results were positive for 21 of 26 PCs (80.8%) and one case of cutaneous leishmaniasis. In 4 of 31 cases, IHC revealed nests (12.9%), which were undetected by conventional histological examination. CONCLUSIONS: This study shows that IHC with the tested antiserum increases the sensitivity of the diagnosis and may be recommended for routine use in EMB analyses of cardiac transplant patients with Chagas disease.


Subject(s)
Humans , Trypanosoma cruzi/immunology , Antibodies, Protozoan/blood , DNA, Protozoan/analysis , Chagas Disease/diagnosis , Endocardium/parasitology , Antibodies, Monoclonal/blood , Biopsy , Immunohistochemistry , Case-Control Studies , Polymerase Chain Reaction , Sensitivity and Specificity , Antibody Formation
7.
Rev. bras. parasitol. vet ; 26(4): 516-520, Oct.-Dec. 2017. tab, graf
Article in English | LILACS | ID: biblio-1042455

ABSTRACT

Abstract Trypanosoma (Duttonella) vivax is an important cause of economic losses among feedlot cattle. These losses are related to the morbidity, mortality, reproductive issues and decreased production. It is known that the clinical signs observed in infections by this protozoon are similar to other hemoparasitosis, which difficult the diagnosis. Therefore, the aim of this study was to detect and molecularly characterize an outbreak of trypanosomiasis caused by T. (D.) vivax in dairy cattle in the municipality of São Miguel Aleixo, state of Sergipe, Brazil. Blood samples from cattle (n = 15) presenting clinical signs compatible with trypanosomiasis were collected and parasitological and molecular evaluated. Among the samples analyzed, 34% (5/15) were positive from blood smears, 60% (9/15) from the buffy coat method and 80% (12/15) from the molecular method. The DNA sequence obtained (659 bp) showed 99% similarity to T. (D.) vivax sequences that are available in the GenBank database. The presence of this protozoon in cattle herds is a problem for producers. Diagnosing trypanosomiasis is problematic because its evolution is similar to that of other parasitic blood diseases. In addition, this is the first report of infection by T. (D.) vivax in cattle in the state of Sergipe, northeastern Brazil.


Resumo Trypanosoma (Duttonella) vivax é responsável por consideráveis perdas econômicas na bovinocultura. Estas perdas estão relacionados à morbidade, mortalidade, problemas reprodutivos e declínio na produção. Sabe-se que os sinais clínicos apresentados em infecções por este protozoário se assemelha a outras hemoparasitoses, dificultando muitas vezes o diagnóstico. Portanto, objetivou-se com este estudo detectar a ocorrência de T. (D.) vivax em bovinos leiteiros no município de São Miguel Aleixo, Estado de Sergipe, Brasil. Para tanto, amostras de sangue (n = 15) foram coletadas e avaliadas através de métodos parasitológicos e moleculares. Do total das amostras analisadas, 34% (5/15) foram positivas no esfregaço sanguíneo, 60% (9/15) pelo método do Buffy Coat, enquanto na biologia molecular 80% (12/15) amplificaram um fragmento de DNA (659 pb) compatível com T. (D.) vivax (GenBank). Em conclusão a presença de T. (D.) vivax nos rebanhos bovinos caracteriza-se como um problema para os pecuaristas, como também para o diagnóstico, uma vez que essa tripanossomíase apresenta evolução semelhante a outras hemoparasitoses. Ademais, este é o primeiro relato de infecção por T. (D.) vivax em bovinos do estado de Sergipe, nordeste do Brasil.


Subject(s)
Animals , Trypanosomiasis, Bovine/epidemiology , Cattle/parasitology , Disease Outbreaks/veterinary , Trypanosoma vivax/isolation & purification , Trypanosomiasis, Bovine/diagnosis , Brazil/epidemiology , Cattle Diseases , DNA, Protozoan/analysis , Trypanosoma vivax/genetics , Dairying
8.
Rev. Soc. Bras. Med. Trop ; 50(3): 350-357, May-June 2017. tab, graf
Article in English | LILACS | ID: biblio-896966

ABSTRACT

Abstract INTRODUCTION: Molecular techniques have been shown to be alternative methods for the accurate detection of infectious and parasitic diseases, such as the leishmaniases. The present study describes the optimization and evaluation of a duplex real-time quantitative PCR (qPCR) protocol developed for the simultaneous detection of Leishmania infantum DNA and sample quality control. METHODS: After preliminary tests with the newly designed TaqMan® probes for the two targets ( L. infantum and glyceraldehyde 3-phosphate dehydrogenase (G3PD) gene), the duplex qPCR protocol was optimized. For the evaluation of the standardized protocol, human blood samples were tested (n=68) and the results were compared to those obtained by reference diagnostic techniques. Statistical analyses included percentage agreement and the Kappa ( k ) coefficient. RESULTS: The detection limit of L. infantum DNA reached 2x10 2 fg (corresponding to ~1 parasite) per µL of blood (ε: 93.9%). The percentage agreement obtained between the duplex VL qPCR and the reference techniques was individually obtained as follows: molecular: 88.3% ( k =0.666; 95% CI 0.437-0.894, good), and serological: 81.7% ( k =0.411; 95% CI 0.125-0.697, moderate). Between the reference techniques, the percentage agreement was 86.7% ( k =0.586; 95% CI 0.332-0.840, moderate). CONCLUSIONS: The new duplex VL qPCR protocol indicated good potential for the accurate, fast, and reliable detection of L. infantum DNA, when applied as a complement to the classical diagnostic tools already available, especially in health or research reference centers.


Subject(s)
Humans , Quality Control , DNA, Protozoan/analysis , Leishmania infantum/genetics , Real-Time Polymerase Chain Reaction/standards , Leishmaniasis, Visceral/diagnosis , Reproducibility of Results , Sensitivity and Specificity
9.
Rev. bras. parasitol. vet ; 26(2): 152-158, Apr.-June 2017. tab, graf
Article in English | LILACS | ID: biblio-899276

ABSTRACT

Abstract Trypanosoma comprises flagellates able to infect several mammalian species and is transmitted by several groups of invertebrates. The order Chiroptera can be infected by the subgenera Herpetosoma, Schizotrypanum, Megatrypanum and Trypanozoon. In this study, we described the diversity of bat trypanosomes and inferred phylogenetic relationships among the trypanosomes from bats caught in Tapajós-Arapiuns Extractive Reserve (Resex) in Pará state. Trypanosomes from bats were isolated by means of hemoculture, and the molecular phylogeny was based on the trypanosome barcode (SSUrDNA V7V8 variable region). A total of 111 bats were caught in the area, belonging to three families (Emballonuridae, Molossidae and Phyllostomidae) and 12 species. The bat trypanosome prevalence, as evaluated through hemoculture, was 9% all positive cultures were cryopreserve (100% of isolation success). Phylogenetic trees grouped nine isolates in T. cruzi marinkellei branch and only one in T. dionisii branch. Studies on bat trypanosome diversity are important for identifying pathogenic species and for generating support for control measures, especially in such areas where humans inhabit the forest with close contact with bat species. In addition, this is the first study in Resex Tapajós-Arapiuns extractive reserve and further studies should be conducted to elucidate the role of these parasites as environmental degradation biomarkers.


Resumo Trypanosoma compreende flagelados capazes de infectar diversas espécies de mamíferos e são transmitidos por diferentes grupos de invertebrados. A ordem Chiroptera pode ser parasitada pelos subgêneros Herpetosoma, Schizotrypanum, Megatrypanum e Trypanozoon. Neste estudo é descrita a diversidade de tripanossomas de morcegos capturados na Reserva Extrativista Tapajós-Arapiuns, no Estado do Pará. Os tripanossomas de morcegos foram isolados através de hemocultura e os estudos filogenéticos baseados na região de barcode de tripanossomas (SSUrDNA V7V8 região variável). Foram capturados 111 morcegos pertencentes a três famílias (Emballonuridae, Molossidae e Phyllostomidae) e 12 espécies. A prevalência dos tripanossomas de morcegos, avaliada por hemocultura, foi de 9% para as culturas positivas e todas foram criopreservadas (100% de eficiência no isolamento). As árvores filogenéticas agruparam nove isolados no ramo de T. cruzi marinkellei e um único isolado de T. dionisii. Estudos sobre a diversidade de tripanossomas de morcegos são importantes para identificar espécies patogênicas e gerar suporte para medidas de controle, principalmente em áreas silvestres com contato entre as populações humanas e de morcegos. Além disso, este foi o primeiro estudo realizado na Resex Tapajós-Arapiuns e novos estudos devem ser conduzidos para elucidar o papel destes parasitas como marcadores de degradação ambiental.


Subject(s)
Animals , Trypanosoma/isolation & purification , Chiroptera/parasitology , Phylogeny , Trypanosoma/classification , Brazil , DNA, Protozoan/analysis , Sequence Analysis, DNA
10.
Mem. Inst. Oswaldo Cruz ; 112(4): 309-318, Apr. 2017. tab, graf
Article in English | LILACS | ID: biblio-841787

ABSTRACT

BACKGROUND Leishmaniases are a serious health problem in southeast Brazil, including the city of Belo Horizonte (BH), Minas Gerais state (MG), where there are high rates of incidence and mortality due to visceral leishmaniases. BH is divided into nine sanitary districts (SD) of which one, the Venda Nova SD, was selected for this study because it has high rates of positivity for canine leishmaniasis and high incidence of human leishmaniasis. OBJECTIVES This study aimed to survey the sand fly fauna in Venda Nova SD from August 2011 to July 2013 and perform a descriptive analysis of the vector population. METHODS The sampling was carried out using automatic HP light traps at all covered areas of the Venda Nova SD, in a total of eighteen light traps. Sampled specimens were identified following Galati (2003), and females were submitted to molecular techniques for the detection and identification of Leishmania DNA. A simple environmental description was done for it area and Kernel estimation was used to infer vector density for each study site. FINDINGS A total of 2,427 sand fly specimens belonging to eight species and five genera were collected of which 95.3% were Lutzomyia longipalpis. The seasonal variation curve was delineated by this species. Lu. longipalpis was the most abundant at all collection points and in all months of the study, and exhibited a natural infection rate of 1.01% for Leishmania infantum and 1.77% for Leishmania braziliensis. MAIN CONCLUSIONS The results show the presence and adaptation of Lu. longipalpis to the anthropic environment of BH and reinforces its role as the main vector of L. infantum in the region.


Subject(s)
Humans , Animals , Male , Female , Dogs , Psychodidae/classification , Psychodidae/parasitology , Leishmaniasis/transmission , DNA, Protozoan/analysis , Insect Vectors/classification , Insect Vectors/parasitology , Leishmania/isolation & purification , Leishmania/classification , Leishmania/genetics , Seasons , Brazil , Population Density
11.
Rev. Inst. Med. Trop. Säo Paulo ; 59: e10, 2017. tab, graf
Article in English | LILACS | ID: biblio-842769

ABSTRACT

ABSTRACT The genetic study of sandfly populations needs to be further explored given the importance of these insects for public health. Were sequenced the NDH4 mitochondrial gene from populations of Nyssomyia neivai from Doutor Camargo, Lobato, Japira, and Porto Rico, municipalities in the State of Paraná, Brazil, to understand the genetic structure and gene flow. Eighty specimens of Ny. Neivai were sequenced, 20 from each municipality, and 269 base pairs were obtained. A total of 27 haplotypes and 28 polymorphic sites were found, along with a haplotypic diversity of 0.80696 and a nucleotide diversity of 0.00567. Haplotype H5, with 33 specimens, was the most common among the four populations. Only haplotypes H5 and H7 were present in all four populations. The population from Doutor Camargo showed the highest genetic diversity, and only this population shared haplotypes with those from the other municipalities. The highest number of haplotypes was sheared with Lobato which also had the highest number of unique haplotypes. This probably occurred because of constant anthropic changes that happened in the environment during the first half of the twentieth century, mainly after 1998. There was no significant correlation between genetic and geographical distances regarding these populations. However, the highest genetic and geographical distances, and the lowest gene flow were observed between Japira and Porto Rico. Geographical distance is a possible barrier between these municipalities through the blocking of haplotype sharing.


Subject(s)
Animals , Female , Genetic Variation/genetics , Insect Vectors/genetics , Psychodidae/genetics , Brazil , DNA, Mitochondrial/analysis , DNA, Protozoan/analysis , Insect Vectors/classification , Leishmaniasis, Cutaneous/transmission , Polymerase Chain Reaction , Psychodidae/classification
12.
Article in English | LILACS | ID: biblio-842783

ABSTRACT

ABSTRACT Rheumatoid arthritis (RA) is a chronic condition that is frequent in patients living in tropical areas exposed to leishmaniasis. RA therapy involves immunosuppressant drugs such as methotrexate (MTX), monoclonal antibodies (mAbs) and prednisone. We report an unusual presentation of cutaneous (CL) or mucocutaneous leishmaniasis (ML) in RA patients from an endemic area of leishmaniasis. A 51-year-old woman noted a cutaneous ulcer on her left ankle during MTX and prednisone RA therapy. Initially diagnosed as a venous stasis ulcer, the aspirate of the injury revealed the presence of Leishmania DNA. A 73-year-old woman presenting non-ulcerated, infiltrated and painful erythematous nodules inside her nostrils while receiving MTX, anti-TNF mAb, and prednisone for RA, had also the aspirate of injuries showing the presence of Leishmania DNA. Both patients healed after the therapy with liposomal amphotericin. The RA therapy has changed to low-dose prednisone, without further reactivation episodes. Both cases suggest that CL or ML can reactivate after administration of an immunosuppressant for RA treatment. Therefore, immunosuppressive treatments for RA should be carefully prescribed in patients from endemic areas or with a history of CL and ML.


Subject(s)
Humans , Female , Middle Aged , Aged , Antirheumatic Agents/adverse effects , Immunosuppressive Agents/adverse effects , Leishmaniasis, Cutaneous/etiology , Leishmania/isolation & purification , Antirheumatic Agents/therapeutic use , Arthritis, Rheumatoid/drug therapy , Arthritis, Rheumatoid/immunology , DNA, Protozoan/analysis , Immunosuppressive Agents/therapeutic use , Leishmaniasis, Cutaneous/diagnosis , Leishmaniasis, Mucocutaneous/immunology , Leishmania/genetics , Recurrence
13.
Rev. bras. parasitol. vet ; 25(1): 61-68, Jan.-Mar. 2016. tab, graf
Article in English | LILACS | ID: lil-777529

ABSTRACT

Abstract This study had the aim of ascertaining the sandfly fauna and possible presence ofLeishmania in these insects, collected in caves in the state of Rondônia, Brazil. Collections were conducted in eight caves located in two different areas of this state. Leishmania in the sandflies collected was detected using the polymerase chain reaction (PCR). This was the first study on sandflies from caves in Rondônia and, among the total of 1,236 individuals collected, 24 species and 10 genera were identified. The speciesEvandromyia georgii was collected for the first time in Rondônia and the most abundant species were Trichophoromyia ubiquitalis with 448 individuals (36.2%), followed by T. octavioi with 283 (22.9%) and E. georgii with 179 (14.5%). For the PCR, 17 pools were analyzed and five pools were positive (forT. auraensis in three pools and for Nyssomyia shawi and N. antunesi in one pool each). The kDNA region was amplified and the presence of Leishmania DNA was confirmed. The sandfly fauna in these caves can be considered diverse in comparison with similar studies in other regions. It may be that some species use caves as a temporary shelter and breeding site, while other species live exclusively in this environment. The detection of LeishmaniaDNA indicates that this pathogen is circulating in cave environments and that further studies are needed in order to ascertain the risks of infection by leishmaniasis in these locations with high touristic potential.


Resumo O objetivo deste estudo foi conhecer a fauna de flebotomíneos, e possível presença de Leishmania nestes insetos, coletados em cavernas do Estado de Rondônia. As coletas foram realizadas em oito cavernas localizadas em duas áreas diferentes do Estado. A detecção de Leishmania nos flebotomíneos foi realizada por reação em cadeia da polimerase (PCR). Este foi o primeiro trabalho com flebotomíneos em cavernas de Rondônia e um total de 1,236 indivíduos foram coletados e identificados em 24 espécies e 10 gêneros.Evandromyia georgii foi coletada pela primeira vez em Rondônia, e as espécies mais abundantes foram Trichophoromyia ubiquitalis com 448 indivíduos (36.2%) seguida por T. octavioi com 283 (22.9%) e E. georgii com 179 (14.5%). No estudo de PCR, 17 pools foram analisados, sendo cinco positivos (T. auraensis - 3, Nyssomyia shawi eN. antunesi - 1 cada). A região do kDNA foi amplificada confirmando a presença de DNA de Leishmania. A fauna de flebotomíneos nestas cavernas foi considerada diversa quando comparada com estudos semelhantes de outras regiões. É possível que algumas espécies utilizem cavernas como abrigo temporário e local de procriação e outras sejam exclusivas deste ambiente. A detecção de DNA de Leishmania indica que este patógeno está circulando no ambiente cavernícola, sendo necessários mais estudos para conhecer o risco de transmissão de leishmanioses nestes locais com alto potencial turístico.


Subject(s)
Animals , Psychodidae/parasitology , Caves/parasitology , Leishmania/isolation & purification , Phlebotomus , Psychodidae/classification , Brazil , DNA, Protozoan/analysis , Insect Vectors , Leishmania/genetics
14.
Mem. Inst. Oswaldo Cruz ; 111(2): 114-119, Feb. 2016. tab, graf
Article in English | LILACS | ID: lil-772617

ABSTRACT

This study aimed to estimate the frequency, associated factors, and molecular characterisation of Entamoeba histolytica, Entamoeba dispar, Entamoeba moshkovskii, andEntamoeba hartmanni infections. We performed a survey (n = 213 subjects) to obtain parasitological, sanitation, and sociodemographic data. Faecal samples were processed through flotation and centrifugation methods.E. histolytica, E. dispar, E. moshkovskii, and E. hartmanni were identified by nested-polymerase chain reaction (PCR). The overall prevalence of infection was 22/213 (10.3%). The infection rate among subjects who drink rainwater collected from roofs in tanks was higher than the rate in subjects who drink desalinated water pumped from wells; similarly, the infection rate among subjects who practice open defecation was significantly higher than that of subjects with latrines. Out of the 22 samples positive for morphologically indistinguishableEntamoeba species, the differentiation by PCR was successful for 21. The species distribution was as follows: 57.1% to E. dispar, 23.8% to E. histolytica, 14.3% toE. histolytica and E. dispar, and 4.8% E. dispar and E. hartmanni. These data suggest a high prevalence of asymptomatic infection by the group of morphologically indistinguishable Entamoeba histolytica/dispar/moshkovskiicomplex and E. hartmanni species. In this context of water scarcity, the sanitary and socioenvironmental characteristics of the region appear to favour transmission.


Subject(s)
Adolescent , Adult , Aged , Aged, 80 and over , Child , Female , Humans , Male , Middle Aged , Young Adult , DNA, Protozoan/analysis , Drinking Water/parasitology , Entamoeba , Entamoebiasis/epidemiology , Feces/parasitology , Molecular Typing/methods , Brazil/epidemiology , Cross-Sectional Studies , Droughts , Entamoeba/classification , Entamoeba/genetics , Polymerase Chain Reaction , Poverty , Prevalence , Water Wells
15.
Arq. bras. oftalmol ; 78(6): 356-358, Nov.-Dec. 2015. tab
Article in English | LILACS | ID: lil-768174

ABSTRACT

ABSTRACT Purpose: To evaluate the ability of real-time quantitative PCR (qPCR) for detectingToxoplasma gondii DNA in the peripheral blood and aqueous humor of patients with toxoplasmic active focal necrotizing retinochoroiditis. Methods: Fifty-five patients with infectious uveitis seen from 2009 to 2013 at the Department of Ophthalmology and Visual Sciences of the Federal University of São Paulo were enrolled in this study. Forty-three patients had toxoplasmic active focal necrotizing retinochoroiditis, and the remaining 12 had non-toxoplasmic infectious uveitis and served as controls. qPCR analysis forT. gondii DNA was performed on the patients' peripheral blood and aqueous humor samples. Results: The qPCR was positive for T. gondii DNA in 37.21% (16/43) of the aqueous humor samples and 2.33% (1/43) of the peripheral blood samples; further, 16.27% (7/43) of the patients had positive results in both their blood and aqueous humor samples. Conclusion: qPCR was able to detect T. gondii DNA in patients with toxoplasmic active focal necrotizing retinochoroiditis in the blood as well as the aqueous humor and can help with the diagnosis of the disease.


RESUMO Objetivo: Analisar o uso do PCR em tempo real (qPCR) na detecção do DNA do T. gondii no sangue periférico e no humor aquoso de pacientes com lesões de retinocoroidite focal, ativa por toxoplasmose. Métodos: Cinquenta e cinco pacientes com uveite infecciosa foram incluídos neste estudo. Os pacientes foram atendidos entre 2009 a 2013, no Departamento de Oftalmologia e Ciências Visuais da Universidade Federal de São Paulo. Quarenta e três pacientes tiveram o diagnóstico de lesões de retinocoroidite focal, ativa por toxoplasmose e, os outros 12 tiveram o diagnóstico de uveíte infecciosa não toxoplásmica e, por isso foram usados como grupo controle. A técnica de qPCR foi utilizada na detecção de DNA do T. gondii em amostras de sangue periférico e humor aquoso. Resultados: O qPCR foi positivo para o DNA do T. gondii em 37,21% (16/43) das amostras de humor aquoso, 2,33% (1/43) nas amostras de sangue periférico e, 16,27% (7/43) em ambas amostras simultaneamente. Conclusão: O qPCR foi capaz de detectar o DNA do T. gondii em pacientes com lesões de retinocoroidite focal, ativa por Toxoplasmose, no sangue bem como, no humor aquoso, podendo ajudar no diagnostico.


Subject(s)
Female , Humans , Male , Aqueous Humor/parasitology , Chorioretinitis/parasitology , Real-Time Polymerase Chain Reaction/methods , Toxoplasma/genetics , Toxoplasmosis, Ocular/parasitology , Uveitis/parasitology , Chorioretinitis/blood , Chorioretinitis/diagnosis , DNA, Protozoan/analysis , DNA, Protozoan/blood , Predictive Value of Tests , Reproducibility of Results , Toxoplasmosis, Ocular/blood , Toxoplasmosis, Ocular/diagnosis , Uveitis/blood
16.
Rev. Soc. Bras. Med. Trop ; 48(5): 555-559, Sept.-Oct. 2015. graf
Article in English | LILACS | ID: lil-763336

ABSTRACT

ABSTRACTINTRODUCTION: In the Americas, mucosal leishmaniasis is primarily associated with infection by Leishmania (Viannia) braziliensis. However, Leishmania (Viannia) guyanensis is another important cause of this disease in the Brazilian Amazon. In this study, we aimed at detecting Leishmaniadeoxyribonucleic acid (DNA) within paraffin-embedded fragments of mucosal tissues, and characterizing the infecting parasite species.METHODS: We evaluated samples collected from 114 patients treated at a reference center in the Brazilian Amazon by polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP) analyses.RESULTS: Direct examination of biopsy imprints detected parasites in 10 of the 114 samples, while evaluation of hematoxylin and eosin-stained slides detected amastigotes in an additional 17 samples. Meanwhile, 31/114 samples (27.2%) were positive for Leishmania spp. kinetoplast deoxyribonucleic acid (kDNA) by PCR analysis. Of these, 17 (54.8%) yielded amplification of the mini-exon PCR target, thereby allowing for PCR-RFLP-based identification. Six of the samples were identified as L. (V.) braziliensis, while the remaining 11 were identified as L. (V.) guyanensis.CONCLUSIONS: The results of this study demonstrate the feasibility of applying molecular techniques for the diagnosis of human parasites within paraffin-embedded tissues. Moreover, our findings confirm that L. (V.) guyanensisis a relevant causative agent of mucosal leishmaniasis in the Brazilian Amazon.


Subject(s)
Female , Humans , Male , Leishmania braziliensis/genetics , Leishmania guyanensis/genetics , Leishmaniasis, Mucocutaneous/parasitology , Mucous Membrane/parasitology , DNA, Protozoan/analysis , Leishmania braziliensis/isolation & purification , Leishmania guyanensis/isolation & purification , Paraffin , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Protozoan Proteins/genetics
17.
Mem. Inst. Oswaldo Cruz ; 110(6): 814-816, Sept. 2015. tab, graf
Article in English | LILACS | ID: lil-763088

ABSTRACT

Currently, there is a trend of an increasing number of Plasmodium vivaxmalaria cases in China that are imported across its Southeast Asia border, especially in the China-Myanmar border area (CMB). To date, little is known about the genetic diversity of P. vivaxin this region. In this paper, we report the first genome sequencing of a P. vivaxisolate (CMB-1) from a vivax malaria patient in CMB. The sequencing data were aligned onto 96.43% of the P. vivaxSalvador I reference strain (Sal I) genome with 7.84-fold coverage as well as onto 98.32% of 14 Sal I chromosomes. Using the de novoassembly approach, we generated 8,541 scaffolds and assembled a total of 27.1 Mb of sequence into CMB-1 scaffolds. Furthermore, we identified all 295 known virgenes, which is the largest subtelomeric multigene family in malaria parasites. These results provide an important foundation for further research onP. vivaxpopulation genetics.


Subject(s)
DNA, Protozoan/analysis , Genome, Protozoan , Plasmodium vivax/genetics , Sequence Analysis, DNA , China/epidemiology , Malaria/epidemiology , Myanmar/epidemiology , Plasmodium vivax/isolation & purification
18.
Rev. Soc. Bras. Med. Trop ; 47(5): 599-606, Sep-Oct/2014. tab, graf
Article in English | LILACS | ID: lil-728889

ABSTRACT

Introduction Leishmania major is the causative agent of zoonotic cutaneous leishmaniasis (ZCL), and great gerbils are the main reservoir hosts in Iran. Abarkouh in central Iran is an emerging focal point for which the reservoir hosts of ZCL are unclear. This research project was designed to detect any Leishmania parasites in different wild rodent species. Methods All rodents captured in 2011 and 2012 from Abarkouh district were identified based on morphological characteristics and by amplification of the rodent cytochrome b (Cyt b) gene. To detect Leishmania infection in rodents, deoxyribonucleic acid (DNA) of each ear was extracted. Internal transcribed spacer-ribosomal deoxyribonucleic acid (ITS-rDNA), microsatellites, kinetoplast deoxyribonucleic acid (kDNA) and cytochrome b genes of Leishmania parasites were amplified by polymerase chain reaction (PCR). Restriction fragment length polymorphism (RFLP) and sequencing were employed to confirm the Leishmania identification. Results Of 68 captured rodents in the region, 55 Rhombomys opimus were identified and nine Leishmania infections (9/55) were found. In addition, eight Meriones libycus and two Tatera indica were sampled, and one of each was confirmed to be infected. Two Meriones persicus and one Mus musculus were sampled with no infection. Conclusions The results showed that all 11 unambiguously positive Leishmania infections were Leishmania major. Only one haplotype of L. major (GenBank access No. EF413075) was found and at least three rodents R. opimus, M. libycus and T. indica—appear to be the main and potential reservoir hosts in this ZCL focus. The reservoir hosts are variable and versatile in small ZCL focal locations. .


Subject(s)
Animals , Haplotypes , Leishmania major/genetics , Leishmaniasis, Cutaneous/veterinary , Rodent Diseases/parasitology , Rodentia/parasitology , Cross-Sectional Studies , DNA, Protozoan/analysis , Disease Reservoirs/parasitology , Genetic Markers , Iran , Leishmania major/isolation & purification , Leishmaniasis, Cutaneous/parasitology , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Rodentia/classification , Zoonoses
19.
Rev. Inst. Med. Trop. Säo Paulo ; 56(5): 391-395, Sep-Oct/2014. tab, graf
Article in English | LILACS | ID: lil-722328

ABSTRACT

Sandflies transmit pathogens of leishmaniasis. The natural infection of sandflies by Leishmania (Viannia) was assessed in municipalities, in the state of Paraná, in Southern Brazil. Sandflies were collected with Falcão and Shannon traps. After dissection in search of flagellates in digestive tubes and identification of the species, female sandflies were submitted to the Multiplex Polymerase Chain Reaction (multiplex PCR) for detection of the fragment of the kDNA of Leishmania (Viannia) and the fragment from the IVS6 cacophony gene region of the phlebotomine insects. The analysis was performed in pools containing seven to 12 guts from females of the same species. A total of 510 female sandflies were analyzed, including nine Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai, and 268 Nyssomyia whitmani. Although none of the females was found naturally infected by flagellates through dissection, the fragment of DNA from Leishmania (Viannia) was shown by multiplex PCR in one sample of Ny. neivai (0.46%) and three samples of Ny. whitmani (1.12%). It was concluded that Ny. neivai and Ny. whitmani are susceptible to Leishmania infection, and that multiplex PCR can be used in epidemiological studies to detect the natural infection of the sandfly vector, because of its sensitivity, specificity and feasibility.


Flebotomíneos transmitem os patógenos das leishmanioses. Foi avaliada a infecção natural de flebotomíneos por Leishmania (Viannia) em municípios do Estado do Paraná, sul do Brasil. Os flebotomíneos foram coletados com armadilhas de Falcão e Shannon. Após dissecação para pesquisa de flagelados no tubo digestório e identificação das espécies, as fêmeas de flebotomíneos foram submetidas a Multiplex Reação em Cadeia da Polimerase (multiplex PCR) para a detecção do fragmento do kDNA de Leishmania (Viannia) e do fragmento do gene IVS6 da cacofonia de flebotomíneos. A análise foi realizada em pools contendo sete a 12 tubos digestórios de fêmeas da mesma espécie. Um total de 510 fêmeas foram analisadas, incluindo nove Migonemyia migonei, 17 Pintomyia fischeri, 216 Nyssomyia neivai e 268 Nyssomyia whitmani. Embora nenhuma fêmea tenha sido encontrada naturalmente infectada com flagelados pela dissecação, o fragmento de DNA de Leishmania (Viannia) foi mostrado por multiplex PCR em uma amostra de Ny. neivai (0,46%) e três amostras de Ny. whitmani (1,12%). Conclui-se que Ny. neivai e Ny. whitmani são suscetíveis à infecção por Leishmania, e que multiplex PCR, devido à sua sensibilidade, especificidade e viabilidade, pode ser utilizada em estudos epidemiológicos para a detecção da infecção natural do inseto vetor.


Subject(s)
Animals , Female , DNA, Protozoan/analysis , Insect Vectors/parasitology , Leishmania/genetics , Psychodidae/parasitology , Brazil , Insect Vectors/classification , Leishmania/isolation & purification , Multiplex Polymerase Chain Reaction , Psychodidae/classification
20.
Mem. Inst. Oswaldo Cruz ; 109(3): 388-390, 06/2014. tab, graf
Article in English | LILACS | ID: lil-711733

ABSTRACT

Giardia duodenalis is one of the most prevalent enteroparasites in children. This parasite produces several clinical manifestations. The aim of this study was to determine the prevalence of genotypes of G. duodenalis causing infection in a region of southeastern Mexico. G. duodenalis cysts were isolated (33/429) from stool samples of children and molecular genotyping was performed by polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) analysis, targeting the triosephosphate isomerase ( tpi ) and glutamate dehydrogenase ( gdh ) genes. The tpi gene was amplified in all of the cyst samples, either for assemblage A (27 samples) or assemblage B (6 samples). RFLP analysis classified the 27 tpi -A amplicons in assemblage A, subgenotype I. Samples classified as assemblage B were further analysed using PCR-RFLP of the gdh gene and identified as assemblage B, subgenotype III. To our knowledge, this is the first report of assemblage B of G. duodenalis in human clinical samples from Mexico.


Subject(s)
Adolescent , Child , Child, Preschool , Female , Humans , Infant , Male , Giardia lamblia/genetics , Giardiasis/parasitology , DNA, Protozoan/analysis , Feces/parasitology , Genotype , Giardia lamblia/isolation & purification , Mexico , Phylogeny , Polymerase Chain Reaction , Polymorphism, Restriction Fragment Length , Sequence Analysis, DNA
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