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1.
Braz J Microbiol ; 54(4): 3221-3230, 2023 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-37653362

RESUMEN

Bats can harbor a diversity of viruses, such as adenovirus. Ten different species of bat adenoviruses (BtAdV A to J) have been previous described worlwide. In Brazil, BtAdV was described in three species of phyllostomid species: Artibeus lituratus, Desmodus rotundus, and Sturnira lilium. There are around 180 bat species in Brazil, with 67% inhabiting the Atlantic Forest, with few information about the circulation of BtAdV in this biome. We aimed to describe the molecular detection and the phylogenetic characterization and suggest a classification of BtAdVs circulating in bats from the Brazilian Atlantic Forest. We collected 382 oral and rectal swabs from 208 bats between 2014-2015 and 2020-2021 from São Paulo, Pernambuco, and Santa Catarina Brazilian states. The adenovirus detection was done by a nested PCR targeting the DNA polymerase gene, and all positive samples were sequenced by the Sanger method. The phylogenetic analyses were based on the amino acid sequences using the MEGA 7 and BEAST software. We obtained 16 positive animals (detection rate 7.7%) belonging to seven bat species: Artibeus lituratus, Carollia perspicillata, Sturnira lilium, Molossus molossus, and the first record of Phyllostomus discolor, Eptesicus diminutus, and Myotis riparius. The phylogenetic analysis based on partial amino acid sequences showed that all obtained AdV sequences belong to the Mastadenovirus genus. We observed a high genetic diversity of BtAdV and identified eleven potential BtAdV species circulating in Brazil (BtAdV K to U). Our results contribute to the epidemiological surveillance of adenovirus, increasing the knowledge about the viral diversity and the distribution of AdV in bats from the Atlantic Forest.


Asunto(s)
Infecciones por Adenoviridae , Quirópteros , Mastadenovirus , Animales , Adenoviridae/genética , Brasil , Filogenia , Variación Genética
2.
Rev Bras Parasitol Vet ; 32(2): e003823, 2023.
Artículo en Inglés | MEDLINE | ID: mdl-37377321

RESUMEN

One hundred and sixty-six cats from two animal shelters were subjected to enzyme-linked immunosorbent assay (ELISA), indirect immunofluorescence antibody test (IFAT), conventional polymerase chain reaction (cPCR), quantitative PCR (qPCR) and parasitological tests (PA) for the diagnosis of Leishmania spp. Among them, 15% (25/166), 53.6% (89/166), 3.6% (06/166) and 1.8% (03/166) were positive by ELISA, IFAT, both PCRs and PA, respectively. The sequencing of ITS-1 PCR amplicons revealed a 100% match with Leishmania infantum. After the Leishmania spp. survey, 12 cats were selected and divided into two groups for clinical, hematological, and biochemical analysis: six L. infantum positive cats (G1) and six Leishmania spp. negative cats (G2). All the cats were negative for feline immunodeficiency virus (FIV) and feline leukemia virus (FeLV). A statistical analysis indicated significantly low platelet counts and significant hyperproteinemia associated with hypoalbuminemia in positive cats (p<0.05). Our results suggest that in endemic areas, cats with clinical signs of feline leishmaniosis (such as skin lesions, weight loss and/or enlarged lymph nodes) and that exhibit hematological and biochemical changes, such as low platelet counts and hyperproteinemia with hypoalbuminemia, should be tested for Leishmania spp. infection.


Asunto(s)
Enfermedades de los Gatos , Hipoalbuminemia , Virus de la Inmunodeficiencia Felina , Leishmania infantum , Leishmaniasis Visceral , Leishmaniasis , Gatos , Animales , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/veterinaria , Hipoalbuminemia/veterinaria , Leishmaniasis/diagnóstico , Leishmaniasis/veterinaria , Leishmaniasis/epidemiología , Virus de la Leucemia Felina , Enfermedades de los Gatos/diagnóstico
3.
Exp Parasitol ; 239: 108294, 2022 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-35679968

RESUMEN

This study evaluated the humoral and cellular response in 100 cats living in an endemic area of visceral leishmaniosis (VL) using the Montenegro Skin Test (MST) and serological diagnosis and compared the MST with other diagnostic techniques. Sixty 60%, (60/100) cats were positive for MST and the diameter of positive skin reactions ranged from 5 to 9 mm. By serological methods, 74% (74/100) and 34% (34/100) had antibodies against Leishmania spp. by Immunofluorescence Antibody Test (IFAT) and Indirect Enzyme-Linked Immunosorbent Assay (ELISA), respectively. Comparing tests, the observed profiles were (1) IFAT (+)/MST (-) = 27 cats, (2) IFAT(-)/MST(+) = 13 cats, (3) IFAT(+)/MST(+) = 47 cats, (4) ELISA(+)/MST(-) = 12 cats, (5) ELISA(-)/MST(+) = 38 cats and (6) ELISA(+)/MST(+) = 22 cats. Through the combination of serological diagnosis and MST, a positivity frequency of 87% (87/100) by IFAT + MST and 72% (72/100) by ELISA + MST was identified in this cat population. Five cats (5%) were positive for Leishmania donovani complex DNA by molecular analysis, and two cats (2%) had Leishmania spp. amastigotes in lymph node smears. Therefore, the agreement between tests was classified as poor for all tests by Kappa index. The IFAT (+)/MST (+) response was the most frequent considering all cats (47%; 47/100); nonetheless, the most frequent immune expression in Polymerase Chain Reaction (PCR)-positive cats was the IFAT (+)/MST (-) profile (80%; 4/5). Five sick and PCR-positive cats, negative for Feline Immunodeficiency Virus (FIV) and Feline Leukemia Virus (FeLV), that PCR sequencing matched 100% with L. donovani complex, all but one were MST negative. These results suggest that cats develop a significant cellular response against infection by parasites of the L. donovani complex, and most PCR and parasitological positive cats may be unable to develop a significant cellular response.


Asunto(s)
Enfermedades de los Gatos , Leishmania infantum , Leishmaniasis Visceral , Animales , Anticuerpos Antiprotozoarios , Antígenos de Protozoos , Enfermedades de los Gatos/diagnóstico , Gatos , Ensayo de Inmunoadsorción Enzimática/veterinaria , Inmunidad Celular , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/epidemiología , Leishmaniasis Visceral/veterinaria , Montenegro , Pruebas Cutáneas
4.
Transbound Emerg Dis ; 69(5): e2863-e2875, 2022 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-35729863

RESUMEN

Bat coronaviruses (Bat-CoVs) represent around 35% of all virus genomes described in bats. Brazil has one of the highest mammal species diversity, with 181 species of bats described so far. However, few Bat-CoV surveillance programmes were carried out in the country. Thus, our aim was to jevaluate the Bat-CoV diversity in the Atlantic Forest, the second biome with the highest number of bat species in Brazil. We analysed 456 oral and rectal swabs and 22 tissue samples from Atlantic Forest bats, detecting Alphacoronavirus in 44 swab samples (9.6%) targeting the RdRp gene from seven different bat species, three of which have never been described as Bat-CoV hosts. Phylogenetic analysis of the amino acid (aa) sequences coding the RdRp gene grouped the sequences obtained in our study with Bat-CoV previously detected in identical or congeneric bat species, belonging to four subgenera, with high aa identity (over 90%). The RdRp gene was also detected in three tissue samples from Diphylla ecaudata and Sturnira lilium, and the partial S gene was successfully sequenced in five tissues and swab samples of D. ecaudata. The phylogenetic analysis based on the partial S gene obtained here grouped the sequence of D. ecaudata with CoV from Desmodus rotundus previously detected in Peru and Brazil, belonging to the Amalacovirus subgenus, with aa identity ranging from 73.6% to 88.8%. Our data reinforce the wide distribution of Coronaviruses in bats from Brazil and the novelty of three bats species as Bat-CoV hosts and the co-circulation of four Alphacoronavirus subgenera in Brazil.


Asunto(s)
Alphacoronavirus , Quirópteros , Infecciones por Coronavirus , Coronavirus , Alphacoronavirus/genética , Aminoácidos/genética , Animales , Brasil/epidemiología , Coronavirus/genética , Infecciones por Coronavirus/veterinaria , Bosques , Variación Genética , Genoma Viral , Filogenia , ARN Polimerasa Dependiente del ARN
5.
Transbound Emerg Dis ; 69(4): 2182-2190, 2022 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-34229362

RESUMEN

Leishmaniasis is a neglected tropical disease that continues to pose a serious public health problem. Albeit dogs have long been held as the major reservoirs of Leishmania infantum, the involvement of domestic cats in the zoonotic cycle of visceral leishmaniasis has gained prominence. Here, 240 cats were evaluated by clinical signs and haematological/biochemical changes compatible with leishmaniasis and were diagnosed by serological, molecular, and parasitological techniques. Thus, four cats naturally infected by L. infantum were submitted to xenodiagnosis. A total of 203 females of Lutzomyia longipalpis were subjected to feeding on four cats, with all females completing the blood meal. Parasitological and molecular assays were carried out to evaluate the presence of L. infantum in the sand flies' midgut. Promastigotes were observed in 10 females (6.5%) that fed on one cat, and L. infantum DNA was detected in 17 (8.4%) females that fed on two cats. Our results strengthen the evidence that naturally infected cats are capable of transmitting L. infantum to sand flies.


Asunto(s)
Enfermedades de los Gatos , Leishmania infantum , Leishmaniasis Visceral , Leishmaniasis , Psychodidae , Animales , Enfermedades de los Gatos/diagnóstico , Gatos , Femenino , Leishmania infantum/genética , Leishmaniasis/veterinaria , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/veterinaria , Masculino , Xenodiagnóstico/veterinaria
6.
Artículo en Inglés | MEDLINE | ID: mdl-33533815

RESUMEN

Visceral leishmaniasis (VL) is a neglected tropical disease caused by the Leishmania infantum parasite. The protozoan is able to infect several domestic and wild mammals. Since the first report on Leishmania spp. infection in horses in South America, leishmaniasis in equids has been highlighted in Brazil. A molecular epidemiological survey was carried out to verify the occurrence of Leishmania spp. DNA in horses and donkeys, in leishmaniases endemic areas in Sao Paulo State, Brazil. To this end, blood samples were obtained from 107 horses and 36 donkeys and subjected to DNA extraction followed by PCR targeting the ITS-1 region. Among the horses and donkeys, 1.87% (2/107) and 8.33% (3/36) were positive by PCR, respectively. The DNA sequencing of the ITS-1 amplification products confirmed L. infantum DNA in these animals. Our results suggest that horses and donkeys from non-VL and VL endemic areas of São Paulo State may be infected by the parasite.


Asunto(s)
Equidae/sangre , Caballos/sangre , Leishmania infantum/genética , Leishmaniasis Visceral/diagnóstico , Animales , Brasil , ADN , Leishmaniasis Visceral/veterinaria , Reacción en Cadena de la Polimerasa
7.
Ciênc. rural (Online) ; 51(10): e20200825, 2021. tab, graf
Artículo en Inglés | LILACS-Express | LILACS, VETINDEX | ID: biblio-1278865

RESUMEN

ABSTRACT: Leishmania infantum causes canine leishmaniasis. Using parasitological and molecular analyses, we identified L. infantum in the reproductive organs of male and female dogs. Using histochemistry, immunohistochemistry, and PCR, we examined tissue samples from the reproductive organs of 8 male dogs and 16 female dogs diagnosed with leishmaniasis. Despite the absence of macroscopic or microscopic lesions in these organs, we observed L. infantum amastigotes in tissue samples from the testis and the uterus. PCR and sequencing of these tissues revealed sequences that matched 100% with L. infantum DNA available at GenBank. The presence of L. infantum amastigotes and DNA in testicular and uterine tissue samples suggested that these organs can harbor the parasite without associated macroscopic or microscopic lesions, and this can be especially important in the vertical and venereal transmission of leishmaniasis in dogs.


RESUMO: Leishmania infantum é agente etiológico da leishmaniose canina. Por meio de análises parasitológicas e moleculares, a presença do parasita foi investigada em órgãos reprodutivos de cães machos e fêmeas. Amostras de tecidos dos órgãos reprodutivos de 8 cães machos e 16 fêmeas diagnosticados com leishmaniose foram avaliadas por histoquímica, imunohistoquímica e PCR. Apesar de não terem sido observadas lesões macroscópicas ou microscópicas nos órgãos reprodutivos desses cães, formas amastigotas de L. infantum foram observadas em amostras teciduais do testículo e útero. A PCR e o sequenciamento do DNA extraído desses tecidos revelaram sequências 100% idênticas a L. infantum depositadas no GenBank. Nossos resultados sugerem que os testículos e o útero podem abrigar o parasita, sem associação com lesões macroscópicas ou microscópicas, o que pode ter uma grande importância na transmissão venérea e vertical da leishmaniose entre cães.

8.
Exp Parasitol ; 218: 107981, 2020 Nov.
Artículo en Inglés | MEDLINE | ID: mdl-32871144

RESUMEN

Phlebotomine sand flies (Diptera: Psychodidae: Phlebotominae) are a group of small insects of great concern for Public Health. These dipterous are intensely studied worldwide due to their involvement in the transmission of several pathogens, mainly Leishmania spp. parasites. Nowadays, the molecular tools have been included in Phlebotomine sand flies studies and has shown to be powerful tools in bioecology studies of these dipterous. Thereby, when molecular approaches are employed, there is a great concern regarding the amount and quality of the DNA obtained for analysis. Here, seven methods of DNA extraction, between commercial kits and in house extraction protocols were evaluated. We considered measure of DNA concentration and purity ratios using a spectrophotometer to check the performance of each protocol. In addition, the quality evaluation of the DNA extracted was performed by endogenous gene PCR on samples. The results of the seven evaluated DNA extraction protocols and their implications are discussed.


Asunto(s)
ADN/aislamiento & purificación , Psychodidae/genética , Análisis de Varianza , Animales , Costos y Análisis de Costo , ADN/análisis , ADN/normas , Electroforesis en Gel de Agar , Femenino , Masculino , Reacción en Cadena de la Polimerasa/métodos , Reacción en Cadena de la Polimerasa/normas , Cloruro de Sodio , Espectrofotometría , Factores de Tiempo
9.
Transbound Emerg Dis ; 67(5): 2013-2019, 2020 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-32162460

RESUMEN

An epidemiological Leishmania spp. and entomological Phlebotomine sandflies survey was performed in cat shelters at leishmaniasis endemic area of Brazil. Blood and conjunctival swab (CS) samples were collected from 94 cats in two animal protection shelters. These samples were subjected to serological tests using the indirect immunofluorescence antibody test (IFAT) and indirect enzyme-linked immunosorbent assay (ELISA) and to molecular test by polymerase chain reaction (PCR). In addition, a Phlebotomine sandflies survey was performed in the same shelters. The analyses revealed a positivity of 31.91% (30/94) through ELISA and 29.79% (28/94) through IFAT. The two serological tests showed a positive association with perfect agreement (k = 0.925). None of the cats were positive by Leishmania spp. DNA. One Lutzomyia (Lutzomyia) longipalpis male was found in one of the cat shelters. The results and the implications of our findings are discussed below.

10.
Rev Bras Parasitol Vet ; 28(2): 303-305, 2019 Jun 13.
Artículo en Inglés | MEDLINE | ID: mdl-31215604

RESUMEN

Leishmania spp. are important agents of human and animal leishmaniases that have an important impact on public health. In this study, we aimed to detect the circulation of Leishmania spp. in cattle from a visceral leishmaniasis non-endemic area of the state of São Paulo, Brazil. DNA was extracted from blood samples from 100 heifers in the municipality of Pirassununga and was amplified using primers specific for the first internal transcriber spacer (ITS1), to assess the presence of trypanosomatids. The assays revealed that one sample presented bands of between 300 and 350 base pairs. In GenBank, this sample matched 100% with Leishmania infantum (314 base pairs). The results suggest that cattle can be infected by Leishmania infantum in Brazil.


Asunto(s)
Enfermedades de los Bovinos/diagnóstico , ADN Espaciador Ribosómico/genética , Leishmania infantum/genética , Leishmaniasis Visceral/veterinaria , Animales , Bovinos , Leishmaniasis Visceral/diagnóstico , Reacción en Cadena de la Polimerasa/veterinaria
11.
Rev Bras Parasitol Vet ; 28(2): 194-202, 2019 Jun 06.
Artículo en Inglés | MEDLINE | ID: mdl-31188942

RESUMEN

The aim of this study was to compare molecular tests used to diagnose Leishmania spp. in dogs with different stages of infection. Blood and conjunctival swab (CS) samples from dogs classified in four clinical stages were subjected to different PCR protocols (13A/13B, MC1/MC2, LITSR/L5.8S and LEISH-1/LEISH-2 primers). To the study, 22.3% (48/215) of dogs were classified as without clinical signs, 67.5% (145/215) stage I (mild disease), 7.0% (15/215) stage II (moderate disease) and 3.2% (7/215) stage III (severe disease). The results showed that in blood samples, 13A/13B detected a significant higher number of positive dogs in stage I (25/145) and in total (42/215) (p≤0.05). However, when CS samples were tested, no difference was observed (p>0.05). On the other hand, in blood samples, MC1/MC2 detected significantly fewer positive dogs classified as without clinical signs (0/48), in stage I (0/145) and in total (1/215) (p≤0.05). Likewise, in CS samples, this primers showed also lower detection (1/215) (p≤0.05). So than, we can conclude that PCR on blood samples with 13A/13B primers has greater capacity to detect positive dogs, mainly at the initial of clinical disease than do other primers and MC1/MC2 are not a good choice to detect Leishmania infantum infection in dogs.


Asunto(s)
Enfermedades de los Perros/diagnóstico , Leishmaniasis Cutánea/veterinaria , Leishmaniasis Visceral/veterinaria , Animales , Brasil/epidemiología , ADN Protozoario/genética , Enfermedades de los Perros/epidemiología , Perros , Leishmania infantum/genética , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Cutánea/epidemiología , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/epidemiología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Índice de Severidad de la Enfermedad
12.
Rev. bras. parasitol. vet ; 28(2): 303-305, Apr.-June 2019. graf
Artículo en Inglés | LILACS | ID: biblio-1042504

RESUMEN

Abstract Leishmania spp. are important agents of human and animal leishmaniases that have an important impact on public health. In this study, we aimed to detect the circulation of Leishmania spp. in cattle from a visceral leishmaniasis non-endemic area of the state of São Paulo, Brazil. DNA was extracted from blood samples from 100 heifers in the municipality of Pirassununga and was amplified using primers specific for the first internal transcriber spacer (ITS1), to assess the presence of trypanosomatids. The assays revealed that one sample presented bands of between 300 and 350 base pairs. In GenBank, this sample matched 100% with Leishmania infantum (314 base pairs). The results suggest that cattle can be infected by Leishmania infantum in Brazil.


Resumo Leishmania spp. são agentes causadores das leishmanioses em humanos e em animais, gerando grande impacto à saúde pública. Este estudo objetivou detectar a circulação de Leishmania spp. em área não endêmica para leishmaniose visceral de São Paulo, Brasil. Foram extraídas amostras de DNA de 100 novilhas da cidade de Pirassununga. Estas amostras foram amplificadas com os iniciadores específicos para tripanosomatídeos Internal Transcriber Spacer 1 (ITS1). Os ensaios revelaram uma amostra com bandas entre 300 e 350 pares de base (pb). A amostra demonstrou 100% de identidade com Leishmania infantum (314 pb). Os resultados sugerem que o gado pode ser infectado por L. infantum no Brasil.


Asunto(s)
Animales , Bovinos , Enfermedades de los Bovinos/diagnóstico , Leishmania infantum/genética , ADN Espaciador Ribosómico/genética , Leishmaniasis Visceral/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Leishmaniasis Visceral/diagnóstico
13.
Rev. bras. parasitol. vet ; 28(2): 194-202, Apr.-June 2019. tab
Artículo en Inglés | LILACS | ID: biblio-1013740

RESUMEN

Abstract The aim of this study was to compare molecular tests used to diagnose Leishmania spp. in dogs with different stages of infection. Blood and conjunctival swab (CS) samples from dogs classified in four clinical stages were subjected to different PCR protocols (13A/13B, MC1/MC2, LITSR/L5.8S and LEISH-1/LEISH-2 primers). To the study, 22.3% (48/215) of dogs were classified as without clinical signs, 67.5% (145/215) stage I (mild disease), 7.0% (15/215) stage II (moderate disease) and 3.2% (7/215) stage III (severe disease). The results showed that in blood samples, 13A/13B detected a significant higher number of positive dogs in stage I (25/145) and in total (42/215) (p≤0.05). However, when CS samples were tested, no difference was observed (p>0.05). On the other hand, in blood samples, MC1/MC2 detected significantly fewer positive dogs classified as without clinical signs (0/48), in stage I (0/145) and in total (1/215) (p≤0.05). Likewise, in CS samples, this primers showed also lower detection (1/215) (p≤0.05). So than, we can conclude that PCR on blood samples with 13A/13B primers has greater capacity to detect positive dogs, mainly at the initial of clinical disease than do other primers and MC1/MC2 are not a good choice to detect Leishmania infantum infection in dogs.


Resumo O objetivo deste estudo foi comparar testes moleculares usados para diagnosticar Leishmania spp., em cães apresentando diferentes estágios de infecção. Amostras de sangue e suabe conjuntival (SC) de cães classificados em quatro estágios clínicos foram submetidas a diferentes PCRs (primers 13A/13B, MC1/MC2, LITSR/L5.8S e LEISH-1/LEISH-2). Para o estudo, 22,3% (48/215) dos cães foram classificados como sem sinais clínicos, 67,5% (145/215) estágio I (doença leve), 7,0% (15/215) estágio II (doença moderada) e 3,2% (7/215) estágio III (doença grave). Os resultados mostraram que, em amostras de sangue, 13A/13B detectou número significativamente maior de cães positivos no estágio I (25/145) e no total (42/215) (p≤0,05). No entanto, quando as amostras de SC foram testadas, nenhuma diferença foi observada (p>0,05). Por outro lado, no sangue, MC1/MC2 detectou significativamente menos cães positivos sem sinais clínicos (0/48), em estágio I (0/145) e no total (1/215) (p≤0,05). Da mesma forma, em amostras de SC, MC1/MC2 também apresentou menor detecção (1/215) (p≤0,05). Assim, a PCR em amostras de sangue com 13A/13B tem maior capacidade de detectar cães positivos, principalmente no início da doença do que outros primers, e o par de primers MC1/MC2 não é uma boa escolha para detectar infecção por Leishmania infantum em cães.


Asunto(s)
Animales , Perros , Leishmaniasis Cutánea/veterinaria , Enfermedades de los Perros/diagnóstico , Leishmaniasis Visceral/veterinaria , Índice de Severidad de la Enfermedad , Brasil/epidemiología , ADN Protozoario/genética , Leishmaniasis Cutánea/diagnóstico , Leishmaniasis Cutánea/epidemiología , Leishmania infantum/genética , Enfermedades de los Perros/epidemiología , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/epidemiología
14.
Exp Parasitol ; 177: 93-97, 2017 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-28438522

RESUMEN

Although some studies have investigated the potential role of cats as a reservoir for Leishmania, their role in the epidemiology of visceral leishmaniasis (VL) is still poorly understood. Molecular diagnostic techniques are an important tool in VL diagnosis, and PCR shows high sensitivity and specificity for Leishmania spp. detection. Quantitative real-time PCR (qPCR) is a method that permits quantitative analysis of a large number of samples, resulting in more sensitive, accurate, and reproducible measurements of specific DNA present in the sample. This study compared real-time PCR (qPCR) and conventional PCR (cPCR) for detection of Leishmania spp. in blood and conjunctival swab (CS) samples of healthy cats from a non-endemic area in the state of São Paulo, Brazil. Of all CS samples, 1.85% (2/108) were positive for Leishmania spp. by both cPCR as qPCR (kappa index = 1), indicating excellent agreement between the two methods. The DNA from the two CS-cPCR- and CS-qPCR-positive samples was further tested with a PCR test amplifying the Leishmania spp. discriminative rRNA internal transcribed spacer 1 (ITS 1), of which one sample generated a 300-350-bp DNA fragment whose size varies according to the Leishmania species. Following sequencing, the fragment showed 100% similarity to a GenBank L. infantum sequence obtained from a cat in Italy. In conclusion, the association of qPCR and CS proved to be effective for detection of Leishmania in cats. Conjunctival swab samples were shown to be a practical and better alternative to blood samples and may be useful in the diagnosis and studies of feline leishmaniasis.


Asunto(s)
Enfermedades de los Gatos/parasitología , Conjuntiva/parasitología , ADN Protozoario/aislamiento & purificación , Leishmania infantum/aislamiento & purificación , Leishmaniasis Visceral/veterinaria , Animales , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/transmisión , Gatos , ADN de Cinetoplasto/aislamiento & purificación , Leishmania infantum/genética , Leishmaniasis Visceral/diagnóstico , Leishmaniasis Visceral/parasitología , Leishmaniasis Visceral/transmisión , Reacción en Cadena de la Polimerasa/veterinaria , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria
15.
Rev Soc Bras Med Trop ; 49(1): 104-6, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-27163572

RESUMEN

INTRODUCTION: Conjunctival swab PCR was evaluated as a tool to diagnose visceral leishmaniasis in dogs. METHODS: Conjunctival swab PCR was compared to indirect immunofluorescence antibody test and blood PCR. RESULTS: Indirect immunofluorescence was significantly correlated with conjunctival swab PCR (p < 0.05), but not with blood PCR (p > 0.05). In addition, conjunctival swab PCR was significantly associated with presence of clinical symptoms (p < 0.05), whereas blood PCR was associated with absence of clinical symptoms (p < 0.05). CONCLUSIONS: Results indicate that conjunctival swab PCR is useful in epidemiological surveys of canine visceral leishmaniasis.


Asunto(s)
Conjuntiva/parasitología , ADN Protozoario/genética , Enfermedades de los Perros/diagnóstico , Leishmania infantum/genética , Leishmaniasis Visceral/veterinaria , Animales , Perros , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Leishmaniasis Visceral/diagnóstico , Valor Predictivo de las Pruebas , Reacción en Cadena en Tiempo Real de la Polimerasa/veterinaria , Sensibilidad y Especificidad
16.
Rev Bras Parasitol Vet ; 24(4): 454-8, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26689180

RESUMEN

We examined the presence of antibodies against the parasites Toxoplasma gondii, Neospora caninum, and Leishmania spp., as well the presence of DNA from Leishmania spp., in dogs from Pirassununga - SP. The seropositivity rate was compared with the animals' originating location. Three hundred seventy-three blood samples from the county's kennel and local veterinary clinics were collected and analyzed. A total of 300 samples were tested for T. gondii and N. caninum using an indirect immunofluorescence antibody test (IFAT); 45% (135/300) were positive for T. gondii and 24.3% (73/300) were positive for N. caninum. Three hundred seventy-three samples were tested for Leishmania spp. using the IFAT. Of these, 4.6% (17/373) were positive. Additionally, 145 samples were tested using a polymerase chain reaction (PCR); of these samples, 0.7% (1/145) was positive. Considering the results, we conclude that these parasites are present in the city of Pirassununga - SP and that the animals have contact with the protozoan. It is therefore necessary to create methods for disease prevention to maintain both animal and human health in regard to leishmaniasis and toxoplasmosis.


Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Perros/inmunología , Leishmania/inmunología , Neospora/inmunología , Toxoplasma/inmunología , Animales , Perros/parasitología , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Estudios Seroepidemiológicos
17.
Rev. bras. parasitol. vet ; 24(4): 454-458, Oct.-Dec. 2015. tab
Artículo en Inglés | LILACS | ID: lil-770317

RESUMEN

Abstract We examined the presence of antibodies against the parasites Toxoplasma gondii, Neospora caninum, and Leishmania spp., as well the presence of DNA from Leishmania spp., in dogs from Pirassununga - SP. The seropositivity rate was compared with the animals’ originating location. Three hundred seventy-three blood samples from the county’s kennel and local veterinary clinics were collected and analyzed. A total of 300 samples were tested for T. gondii and N. caninum using an indirect immunofluorescence antibody test (IFAT); 45% (135/300) were positive for T. gondii and 24.3% (73/300) were positive for N. caninum. Three hundred seventy-three samples were tested for Leishmania spp. using the IFAT. Of these, 4.6% (17/373) were positive. Additionally, 145 samples were tested using a polymerase chain reaction (PCR); of these samples, 0.7% (1/145) was positive. Considering the results, we conclude that these parasites are present in the city of Pirassununga - SP and that the animals have contact with the protozoan. It is therefore necessary to create methods for disease prevention to maintain both animal and human health in regard to leishmaniasis and toxoplasmosis.


Resumo Avaliou-se a presença de anticorpos contra Toxoplasma gondii, Neospora caninum e Leishmania spp.; assim como a presença de DNA de Leishmania spp. em cães de Pirassununga-SP, e associou-se sua soropositividade ao local de origem dos animais. Foram coletadas 373 amostras de sangue do canil municipal e de clínicas veterinárias locais, que foram analisadas pelo teste de Imunofluorescência Indireta (RIFI). Do total, 300 amostras foram testadas para T. gondii e N. caninum, das quais 45% (135/300) foram positivas para T. gondii e 24,3% (73/300) para N. caninum. Para Leishmania spp. foram avaliadas 373 amostras pela RIFI, sendo 4,6% (17/373) positivas. Adicionalmente, 145 amostras foram testadas utilizando-se a PCR e, dessas amostras, 0,7% (1/145) foi positiva. Considerando-se os resultados, pode-se concluir que esses parasitos estão presentes na cidade de Pirassununga - SP e que os animais tiveram contato com os protozoários. Faz-se, dessa forma, necessária a divulgação de meios de prevenção às doenças, com o intuito de manter o controle sobre as mesmas, tanto na saúde animal quanto na saúde humana, em relação à leishmaniose e à toxoplasmose.


Asunto(s)
Animales , Toxoplasma/inmunología , Anticuerpos Antiprotozoarios/sangre , Neospora/inmunología , Perros/inmunología , Leishmania/inmunología , Estudios Seroepidemiológicos , Reacción en Cadena de la Polimerasa/veterinaria , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Perros/parasitología
18.
Rev Bras Parasitol Vet ; 24(2): 220-2, 2015.
Artículo en Inglés | MEDLINE | ID: mdl-26154963

RESUMEN

The relevance of the dog as a source of visceral leishmaniasis infection is known, but the role of cats as reservoir hosts for leishmaniasis is not yet fully clear. This study assessed the efficacy of conjunctival swab PCR (CS-PCR) in the detection of cats infected by Leishmania spp. The results were seven (13.5%) cats positive for Leishmania spp. in the PCR, in 52 cats tested from Pirassunuga-SP and Ilha Solteira-SP. From the city of Pirassununga - SP 28.6% (2/7) were positive and from the city of Ilha Solteira - SP 11.1% (5/45) were positive. The results showed that CS-PCR was capable of detecting cats infected by this protozoan. Conjunctival swab samples proved easier to perform in cats, which might facilitate studies on the frequency and distribution of feline leishmaniasis.


Asunto(s)
Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/parasitología , Conjuntiva/parasitología , ADN Protozoario/análisis , Leishmania/genética , Leishmania/aislamiento & purificación , Leishmaniasis/veterinaria , Animales , Gatos , Femenino , Leishmaniasis/diagnóstico , Masculino , Reacción en Cadena de la Polimerasa
19.
Rev. Bras. Parasitol. Vet. (Online) ; 24(2): 220-222, n/2015n/2015.
Artículo en Inglés | LILACS, VETINDEX | ID: biblio-1487851

RESUMEN

The relevance of the dog as a source of visceral leishmaniasis infection is known, but the role of cats as reservoir hosts for leishmaniasis is not yet fully clear. This study assessed the efficacy of conjunctival swab PCR (CS-PCR) in the detection of cats infected by Leishmania spp. The results were seven (13.5%) cats positive for Leishmania spp. in the PCR, in 52 cats tested from Pirassunuga-SP and Ilha Solteira-SP. From the city of Pirassununga SP 28.6% (2/7) were positive and from the city of Ilha Solteira SP 11.1% (5/45) were positive. The results showed that CS-PCR was capable of detecting cats infected by this protozoan. Conjunctival swab samples proved easier to perform in cats, which might facilitate studies on the frequency and distribution of feline leishmaniasis.


A importância do cão como fonte de infecção da leishmaniose visceral já é conhecida, mas o papel dos gatos como reservatórios das leishmanioses ainda não está totalmente esclarecido. O presente estudo avaliou a eficácia da PCR de suabe conjuntival (PCR-SC) na detecção de gatos infectados por Leishmania spp. Foram encontrados sete (13,5%) gatos positivos para Leishmania spp. na PCR de suabe conjuntival, dentre 52 animais de Pirassununga - SP e Ilha Solteira - SP testados. Sendo positivos 28,6% (02/07) dos gatos do município de Pirassununga e 11,1% (5/45) dos gatos do município de Ilha Solteira. Os resultados demonstraram que o suabe de conjuntiva ocular foi capaz de detectar gatos infectados por esse protozoário. A coleta de amostras da conjuntiva mostrou ser um método simples, menos invasivo e pouco estressante para os gatos e seus proprietários, o que pode facilitar estudos sobre a frequência e distribuição da leishmaniose felina.


Asunto(s)
Animales , Gatos , Enfermedades de los Gatos/diagnóstico , Enfermedades de los Gatos/parasitología , Leishmania/genética , Leishmania/aislamiento & purificación , Leishmaniasis/diagnóstico , Leishmaniasis/veterinaria , Reacción en Cadena de la Polimerasa
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