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ABSTRACT: We describe an outbreak of leishmaniasis in seven guinea pigs (Cavia porcellus) in which nodular ulcerated skin lesions of varying sizes were observed in the nasal cavity, upper lip, pinnae, vulva, and periarticular region of the limbs. Cytologic exam of collected samples of the lesions in the auricle of one of the animals revealed macrophages containing parasitophorous vacuoles of approximately 4.0μm in diameter in their cytoplasm with morphology suggestive of Leishmania sp. Although skin lesions spontaneously regressed in two of the Guinea pigs, only one survived. All six animals that died were necropsied. Grossly, all animals showed bloody nodular cutaneous lesions with crusts. One of the guinea pigs had distended dark red and firm lungs. Histopathology of the skin lesions revealed histiocytic interstitial acanthotic dermatitis associated with a myriad of Leishmania organisms within macrophages cytoplasm. In the lung, the lesions were characteristic of broncho-interstitial pneumonia with focal infiltrates of neutrophils, epithelioid macrophages, and multinucleated giant cells containing 2µm basophilic amastigotes with morphology compatible with Leishmania spp. A focal granulomatous lesion ,associated with the causal agent in the lung is a novel description of leishmaniasis in guinea pigs caused by L. enriettii. The polymerase chain reaction (PCR) technique with mini-exon primer performed in samples of lesions from two affected guinea pigs was positive and equal to the reference strain, identifying Leishmania enriettii. The cytological, macroscopic, and histological lesions associated with the PCR technique allowed the diagnosis of leishmaniasis and the identification of the specie L. enriettii.

RESUMO: Descrevemos um surto de leishmaniose em sete cobaias (Cavia porcellus), com lesões cutâneas nodulares ulceradas de tamanhos variados observadas na cavidade nasal, lábio superior, pavilhões auriculares, vulva e região periarticular dos membros. No exame citológico foram encontrados macrófagos contendo vacúolos parasitóforos no citoplasma de aproximadamente 4.0μm em diâmetro com morfologia sugestiva de Leishmania sp. Apesar de regressão espontânea das lesões cutâneas terem ocorrido em duas das sete cobaias, apenas um sobreviveu. Seis dos sete animais afetados morreram e foram necropsiados. Macroscopicamente, todos os animais apresentaram lesões cutâneas nodulares, crostosas e sanguinolentas. Uma das cobaias tinha pulmões vermelho-escuros, distendidos e firmes. A histopatologia das lesões cutâneas revelou dermatite acantótica intersticial histiocítica associada a miríades de organismos de Leishmania no citoplasma de macrófagos. Nos pulmões as lesões eram características de pneumonia bronco-intersticial com infiltrado focal de neutrófilos, eosinófilos, macrófagos epitelioides e células gigantes multinucleadas contendo amastigotas basofílicos de 2µm com morfologia compatível com Leishmania spp. Lesões granulomatosas focais associadas ao agente no pulmão são um achado inédito na leishmaniose causada por L. enriettii em cobaias. A técnica de reação em cadeia da polimerase (PCR) com primer mini-exon realizada em amostras de lesões de duas cobaias afetadas foi positiva, identificando Leishmania enriettii. Os aspectos macroscópicos, citológicos, e histológicos associados à técnica da (PCR), permitiram o diagnóstico da leishmaniose e a identificação da espécie L. enriettii.

https://doi.org/10.1590/1678-5150-pvb-7241

First report of putative Leishmania RNA virus 2 (LRV2) in Leishmania infantum strains from canine and human visceral leishmaniasis cases in the southeast of Brazil
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Rêgo, Felipe Dutra

; da Silva, Eduardo Sérgio

; Lopes, Valeriana Valadares

; Teixeira-Neto, Rafael Gonçalves

; Belo, Vinícius Silva

; Fonseca Júnior, Antônio Augusto

; Pereira, Diego Andrade

; Pena, Heber Paulino

; Laurenti, Márcia Dalastra

; Araújo, Gabriela V

; da Matta, Vânia Lúcia Ribeiro

; Chouman, Islam Hussein

; Burrin, Thainá Bergantin

; Sandoval, Carmen M

; Barrouin-Melo, Stella Maria

; de Pinho, Flaviane Alves

; de Andrade, Hélida Monteiro

; Nunes, Ramon Vieira

; Gontijo, Célia Maria Ferreira

; Soccol, Vanete Thomaz

; Klocek, Donnamae

; Grybchuk, Danyil

; Macedo, Diego Henrique

; do Monte-Neto, Rubens Lima

; Yurchenko, Vyacheslav

; Soares, Rodrigo Pedro

Memórias do Instituto Oswaldo Cruz

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2023, Volume 118 elocation e230071

Abstract: En | Text: En | PDF: En

BACKGROUND Leishmania RNA virus 1 (LRV1) is commonly found in South American Leishmania parasites belonging to the subgenus Viannia, whereas Leishmania RNA virus 2 (LRV2) was previously thought to be restricted to the Old-World pathogens of the subgenus Leishmania. OBJECTIVES In this study, we investigated the presence of LRV2 in strains of Leishmania (L.) infantum, the causative agent of visceral leishmaniasis (VL), originating from different hosts, clinical forms, and geographical regions. METHODS A total of seventy-one isolates were screened for LRV2 using semi-nested reverse transcription-polymerase chain reaction (RT-PCR) targeting the RNA-dependent RNA polymerase (RdRp) gene. FINDINGS We detected LRV2 in two L. infantum isolates (CUR268 and HP-EMO) from canine and human cases, respectively. MAIN CONCLUSIONS To the best of our knowledge, this is the first detection of LRV2 in the New World.

https://doi.org/10.1590/0074-02760230071

GENETIC DIVERSITY OF Lutzomyia (Nyssomyia) intermedia IN AN ENDEMIC AREA OF AMERICAN CUTANEOUS LEISHMANIASIS, STATE OF PARANÁ, BRAZIL
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GONÇALVES, André Luiz

; CASTRO, Edilene Alcântara de

; LUZ, Ennio

; Cancio FENDRICH, Ricardo

; Araújo de SOUZA, Nataly

; THOMAZ-SOCCOL, Vanete

Acta Biológica Colombiana

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dec 2021, Volume 26 N. 3 Pages 365 - 373

Abstract: Es En | Text: Es En | PDF: Es | PDF: En

RESUMEN Lutzomyia intermedia (Diptera: Psychodidae) es uno de los principales vectores que participan en la transmisión de leishmaniasis cutánea americana (LCA) en la región Neotropical. A pesar de que aún los estudios genéticos que involucran a este taxón son incipientes, tienen una gran importancia para comprender el nivel de variabilidad de las diferentes poblaciones y sus implicaciones en su papel vectorial. El objetivo de este estudio fue determinar el nivel de diversidad genética de L. intermedia presente en el Valle del Río Ribeira, área de transmisión de LCA en el estado de Paraná, Brasil, mediante RAPD (ADN polimórfico amplificado aleatoriamente). Los flebótomos fueron recolectados en los municipios Cerro Azul (nueva área de transmisión de LCA) y Adrianópolis (área endémica de LCA), donde fueron capturados en ambientes residenciales, en el peridomicilio y en el bosque. Dos de los marcadores utilizados permitieron estimar el polimorfismo en las poblaciones estudiadas con la obtención de 40 genotipos, la mayoría de ellos en el peridomicilio. El dendrograma generado por el análisis con el cebador A10 mostró diferentes grados de similitud, lo que sugiere que puede haber flujo gènico en las poblaciones. El Análisis de Coordenadas Principales (PCO) con el cebador A2 fue útil para agrupar L. intermedia según su origen ecológico y geográfico. No hubo distinción entre los linajes que componen el complejo L. intermedia. Los resultados de este estudio, con el registro de gran diversidad genotipica en L. intermedia, pueden contribuir a explicar el mantenimiento del ciclo biológico de Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) en la región.

ABSTRACT Lutzomyia intermedia (Diptera: Psychodidae) features as one of the main vectors that are involved in the transmission of American cutaneous leishmaniasis (ACL) in the Neotropical region. However, genetic studies involving this taxon are still incipient and important for understanding the level of variability of different populations, their role, and implications as vectors. The aim of this study was to determine the level of genetic diversity of L. intermedia present in the Ribeira River Valley, an area of ACL transmission in the state of Paraná, Brazil, through the Random amplified polymorphic DNA (RAPD). Two municipalities were chosen to collect sand flies: Cerro Azul (new transmission area of the ACL) and Adrianópolis (endemic area of the ACL). The insects were captured in the house, in the peridomicile and in the wild (forest). Two of the used markers made it possible to estimate the polymorphism of the studied populations, resulting in 40 genotypes, most of them from peridomicile. The dendrogram generated by the analysis with the primer A10 showed different degrees of similarity, suggesting that there may be gene flow in the studied populations. The Principal Coordinate Analysis (PCO) with the A2 primer, was useful in grouping L. intermedia according to its ecological and geographical origin. There was no distinction between the lineages composing the L. intermedia complex. The results of this study, with the record of great genotypic diversity in L. intermedia, may contribute to explain the maintenance of the life cycle of Leishmania braziliensis (Kinetoplastida: Trypanosomatidae) in the region.

https://doi.org/10.15446/abc.v26n3.84619 9 downloads

Correlation of maternal concentrations of plasma testosterone with fetal sex in horses
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Busato, Eduarda Maciel

; Weiss, Romildo Romualdo

; Abreu, Ana Claudia Machinski Rangel de

; Bergstein-Galan, Tácia Gomes

; Marcondes, Felipe Augusto Beckert

; Kozicki, Luiz Ernandes

; Soccol, Vanete Thomaz

; Dornbusch, Peterson Triches

Ciência Rural

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2021, Volume 51 N. 2 elocation e20200237

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

RESUMO: Os objetivos do estudo foram avaliar a correlação do sexo fetal com as concentrações plasmáticas de testosterona entre o 5° e o 8º mês de gestação na égua e verificar a aplicabilidade deste exame para a predição do sexo fetal. Amostras de sangue foram coletadas de 21 éguas, com intervalos de 30 dias, entre 150 e 240 dias de gestação. A testosterona plasmática foi determinada por radioimunoensaio e o sexo dos potros foi confirmado ao nascimento. Os valores de testosterona materna foram superiores nas éguas gestando fetos fêmeas aos cinco e oito meses (P< 0.05). Através da análise da curva ROC (receiver operating characteristic) foram determinados valores limites de 35,5 pg/mL e 40 pg/mL para o 5º e o 8° mês, respectivamente. Éguas com testosterona plasmática igual ou acima dos valores limites foram preditas como gestando fêmeas e éguas com testosterona plasmática abaixo dos valores limites foram preditas como gestando machos. Aos cinco meses, os valores preditivos para fetos machos e fêmeas foram 70% e 88,9%, respectivamente; as taxas de detecção foram 87,5% e 72,7% e a acurácia total do exame foi de 78,9%. Aos oito meses, os valores preditivos para fetos machos e fêmeas foram 80% e 90%, respectivamente; as taxas de detecção foram 88,9% e 81,8% e a acurácia total do exame foi de 85%. Conclui-se que houve correlação entre o sexo fetal e as concentrações de testosterona plasmática em éguas prenhes. A predição do sexo fetal baseada nas concentrações plasmáticas de testosterona materna pode ser realizada aos cinco e oito meses de gestação com 78,9% e 85% de acurácia, respectivamente.

ABSTRACT: The objectives of this study were to evaluate the correlation of fetal sex and plasma testosterone concentrations between the 5th and 8th months of pregnancy in mares and to verify the applicability of this test to predict fetal sex. Blood samples were collected from 21 mares at 30-day intervals of between 150 and 240 days of pregnancy. Plasma testosterone was determined by radioimmunoassay and the sex of the foals confirmed at birth. The levels of maternal testosterone were higher in mares carrying female fetuses at months 5 and 8 (P < 0.05). Limit values were determined by analyzing the receiver operating characteristic (ROC) estimates: 35.5 pg/mL and 40 pg/mL for the 5th and 8th month, respectively. For the mares with plasma testosterone values equal to or above the threshold, gestation of female foals was predicted, and for those with plasma testosterone below the threshold values pregnancy of male foals was predicted. In the 5th month, the predictive values for male and female fetuses were 70% and 88.9%, respectively; the detection rates were 87.5% and 72.7%, and the total accuracy of the examination was 78.9%. In the 8th month, the predictive values for male and female fetuses were 80% and 90%, respectively; the detection rates were 88.9% and 81.8%, and the total accuracy of the examination was 85%. It was concluded that there was a correlation between fetal sex and plasma testosterone concentrations in pregnant mares. Prediction of fetal sex based on plasma concentrations of maternal testosterone can be performed in months 5 and 8 with 78.9% and 85% accuracy, respectively.

https://doi.org/10.1590/0103-8478cr20200237 818 downloads

Formulation and Validation of Recombinant Antigens CFP10 and ESAT6 for Tuberculosis Diagnosis
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Araujo, Ludmilla Dela Coletta Troiano

; Rodriguez-Fernández, Daniel Ernesto

; Karp, Susan Grace

; Marquez, Ellen de Souza

; Santos, Aline Cristina dos

; Hospinal-Santiani, Manuel

; Boschero, Raphael Aparecido

; Ramos, Eliezer Lucas Pires

; Thomaz-Soccol, Vanete

Brazilian Archives of Biology and Technology

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2021, Volume 64 N. spe elocation e21210127

Abstract: En | Text: En | PDF: En

Abstract The rapid and accurate diagnosis of tuberculosis (TB), especially considering limited resources, is still a challenge. Development of new methodologies and tests are needed to overcome several disadvantages of the available standard tests. We evaluated the diagnostic potential of two antigens specific for Mycobacterium tuberculosis, the CFP10 and ESAT6 recombinant proteins, and developed stable formulations thereof. Sensitivity and specificity of the delayed-type hypersensitivity (DTH) skin testing and the induction of gamma interferon production (IFN-γ) by lymphocytes, as a non-invasive test, were evaluated using the CFP10 and ESAT6 protein formulations. The recombinant proteins produced by our group presented a high DTH response and the ability to differentiate between tuberculosis infection, BCG vaccination, and the contact with non-tuberculous mycobacteria (NTM). The production of IFN-γ by stimulation with individual and combined proteins was detected in a panel of 40 individuals and showed a specificity of 100% and a sensitivity of 90% when the two proteins were used together. Lyophilized formulations were stable under all conditions, while soluble formulations were stable under freezing at -20 ºC and -80 ºC. The proposed formulations containing the ESAT6 and CFP10 recombinant antigens constitute satisfactory tools for TB testing, suitable to be developed and implemented in a large-scale trial.

https://doi.org/10.1590/1678-4324-75years-2021210127 122 downloads

A Review on COVID-19 Diagnosis Tests Approved for Use in Brazil and the Impact on Pandemic Control
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Vedova-Costa, Jean Michel Dela

; Ramos, Eliézer Lucas Pires

; Boschero, Raphael Aparecido

; Ferreira, Gabriela Nascimento

; Soccol, Vanete Thomaz

; Santiani, Manuel Hospinal

; Pacce, Violetta Dias

; Lustosa, Bruno Paulo Rodrigues

; Vicente, Vânia Aparecida

; Soccol, Carlos Ricardo

Brazilian Archives of Biology and Technology

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2021, Volume 64 N. spe elocation e21200147

Abstract: En | Text: En | PDF: En

Abstract With the COVID-19 pandemic, many diagnostic tests (molecular or immunological) were rapidly standardised, given the urgency of the situation, many are still in the process of being validated. The main objective of this study was to review the aspects of the diagnostic kits approved in Brazil and their application in the different federative units to gather epidemiological information. In order to achieve these objectives, a survey was carried out on the data available at the regulatory agency (ANVISA) and in the literature. The main countries that have registered products in Brazil are China (51.4%), Brazil (16.6%), South Korea (9.2%), USA (8.8%) and Germany (3.6%). The methodologies of these products are based on the detection of nucleic-acid (15.8%), antigen (13%) and antibody (71.2%). In the immunological tests, it was verified that the sensitivity ranged from 55 to 100% and the specificity from 80 to 100%. The percentage of cases in the samples tested in Brazil is elevated in almost all federative units since eight states showed 40% of positive cases in tested samples, while 18 states displayed between 20 and 40%. In conclusion, this review showed that Brazil is dependent on external technology to respond to pandemics, epidemics and endemics disease and needs to improve its biotechnological scheme to solve further diseases outbreaks.

https://doi.org/10.1590/1678-4324-75years-2021200147 329 downloads

Guinea Pigs Naturally Infected by Leishmania enriettii: Clinical Analyses, Parasite Isolation and Identification
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Soccol, Vanete Thomaz

; Castro, Edilene Alcântara de

; Reifur, Larissa

; Teixeira, Valéria Natascha

; Lange, Rogério Ribas

; Luz, Ennio

Brazilian Archives of Biology and Technology

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2021, Volume 64 N. spe elocation e21210095

Abstract: En | Text: En | PDF: En

Abstract Leishmania enriettii has only been found in Curitiba metropolitan region, southern Brazil were it was first observed in Cavia porcellus from the vivarium of Instituto de Biologia e Pesquisas Tecnológicas (IBPT - today named TECPAR) by Medina, 1944. Despite more than a half century from its discovery and several research articles on this species, the natural clinical signs in guinea pigs and the parasite genetic variability is still unclear. The aims of this study were to describe the clinical features, investigate the potential wild reservoirs and, in addition, we intended to understand the polymorphism trait of the species. We analyzed 26 naturally infected guinea pigs from eight Paraná state cities. All animals showed lesions compatible with leishmaniosis, such as skin nodules or ulcers on body extremities. Direct examination of the lesion samples obtained by fine-needle aspiration or punch biopsy was conducted followed by isolation and identification of parasite DNA by random amplification of polymorphic DNA (RAPD)-PCR. Through the direct exam, a large number of intracellular amastigote forms were observed in the lesions. Different strains of the parasite, isolated from the 26 animals, were grouped in 5 clusters of approximately 65% similarity. We looked for L. enriettii in other potential reservoir hosts but the parasite was not observed. These results confirm that distinct strains of L. enriettii circulate in guinea pigs from Paraná state, more specifically in the Atlantic forest region, where we believe it serves as the center for dispersion of the species.

https://doi.org/10.1590/1678-4324-75years-2021210095 1 downloads

A comparative study of extraction techniques for maximum recovery of bioactive compounds from Ganoderma lucidum spores
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Saavedra Plazas, Diana Carolina

; Soccol, Carlos Ricardo

; Noseda, Miguel Daniel

; de Andrade Tanobe, Valcineide Oliveira

; Marin, Oranys

; Karp, Susan Grace

; de Melo Pereira, Gilberto Vinicius

; de Carvalho, Júlio Cesar

; Soccol, Vanete Thomaz

Revista Colombiana de Ciencias Químico - Farmacéuticas

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apr 2020, Volume 49 N. 1 Pages 70 - 88

Abstract: Es En | Text: Es En | PDF: Es | PDF: En

RESUMEN El objetivo de este estudio fue evaluar varios métodos de ruptura de las esporas de Ganoderma lucidum y extraer sus propiedades bioactivas. Para este propósito se evaluaron diferentes técnicas de rompimiento como: la maceración manual (RM), la maceración con esferas de diversos materiales (BR) y la exposición a microondas junto la maceración de las esporas con esferas de acero/cromo (MBR1). La ruptura de las esporas fue evaluada por espectroscopia UV-Vis, la cual mostró que las vibraciones 2955, 1642, 1240, 1080 y 1746 cm-1 correspondieron a cambios estructurales en las paredes de las esporas. El extracto MBR1 presento el mayor contenido de carbohidratos (19,80 mg.g-1) y polifenoles (2,21 mg.g-1), mientras que el extracto BR tuvo una mayor actividad antioxidante (57,22% Inb DPPH). Los extractos MBR1 y BR también presentaron en el análisis de monosacáridos un 62,2 y 73,5% de contenido glucosa. Como conclusión la mejor metodología para extraer biomoléculas de las paredes de las esporas de G. lucidum fueron el tratamiento térmico con microondas y la ruptura de las paredes con esferas de acero/cromo, porque este proceso permitió la extracción de una mayor cantidad de carbohidratos con posibles propiedades antioxidantes.

SUMMARY This study aimed at evaluating effective methods for breaking the hard and insoluble spores of Ganoderma lucidum to recover functional biomolecules. Rupture techniques were evaluated such as manual maceration (RM), maceration with spheres of various materials (BR), and microwave exposure plus maceration with steel/ chrome spheres (MBR1). Spore rupture was evaluated using UV-Vis spectroscopy, which showed vibrations of 2955, 1642, 1240, 1080 and 1746 cm-1 corresponding to changes in spore walls. The MBR1 extract contained the largest amounts of carbohydrates (19.80 mg.g-1 spores) and polyphenols (2.21 mg.g-1 spores), whereas the BR extract had higher antioxidant activity (57.22%Inb DPPH). The MBR1 and BR extracts contained 62.2 and 73.5% glucose, respectively. Both methods also involved significant extraction of carbohydrates and proteins. The best way to extract biomolecules from spore walls is to perform a microwave heat treatment and break the walls with steel/chrome spheres; this produces large quantities of carbohydrates with antioxidant properties.

https://doi.org/10.15446/rcciquifa.v49n1.84456 390 downloads

Autochthonous canine visceral leishmaniasis cases occur in Paraná state since 2012: isolation and identification of Leishmania infantum
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Dias, Renata Cristina Ferreira

; Pasquali, Aline Kuhn Sbruzzi

; Thomaz-Soccol, Vanete

; Pozzolo, Eliane Maria

; Chiyo, Luciana

; Alban, Silvana Maria

; Fendrich, Ricardo Cancio

; Almeida, Rômulo Augusto Andrade de

; Ferreira, Fernanda Pinto

; Caldart, Eloiza Teles

; Freire, Roberta Lemos

; Mitsuka-Breganó, Regina

; Bisetto Júnior, Alceu

; Navarro, Italmar Teodorico

Revista Brasileira de Parasitologia Veterinária

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2020, Volume 29 N. 1 elocation e009819

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo O objetivo deste estudo foi confirmar a emergência da leishmaniose visceral canina em Foz do Iguaçu próximo à fronteira com a Argentina e ao Paraguai, por meio do isolamento e identificação molecular de Leishmania infantum. Em um primeiro estágio de coleta de animais (2012), três amostras de aspirados de linfonodos e 46 camadas leucocitárias foram obtidas de cães soropositivos para leishmaniose. Em um segundo estágio de coleta (2013), foram coletadas amostras de camada leucocitária de 376 cães de 20 localidades próximas à fronteira com o Paraguai, rio Paraná, centro e periferia da cidade. Seis isolados foram obtidos, quatro da primeira etapa (4/49) e dois da segunda etapa (2/376); estes isolados foram submetidos à amplificação com iniciadores K26F/R, e a análise de sua sequência confirmou a espécie como L. infantum. A autoctonia dos casos foi confirmada, pois 100% dos cães nunca haviam saído da cidade. O estudo confirma a emergência de leishmaniose visceral canina no Paraná com identificação de L. infantum em cães da cidade de Foz do Iguaçu. Assim, medidas de controle devem ser elaboradas e implementadas por órgãos públicos e instituições de pesquisa do Brasil, Argentina e Paraguai em parceria com o objetivo de controlar a disseminação de zoonoses e os casos humanos de LV.

Abstract The aim of this study was to confirm the emergence of canine visceral leishmaniasis among dogs in Foz do Iguaçu. The disease was diagnosed through the isolation and molecular identification of Leishmania infantum. In the first sample collection stage (2012), three lymph node aspirates and 46 buffy coat samples were obtained mostly from the dogs that were seroreagents for leishmaniasis. In the second sample collection stage (2013), the buffy coat samples were collected from 376 dogs located close to Paraguay, Paraná river, center and peripheral parts of the city. The DNA from the six isolates, four from the first sampling stage (4/49) and two from the second sampling stage (2/376), was subjected to polymerase chain reaction using the K26F/R primers. The isolate was confirmed as L. infantum by sequencing. As none of the dogs had ever left the city, the isolates were confirmed as autochthonous. Further, the study confirmed the emergence of canine visceral leishmaniasis in Paraná through the identification of L. infantum among dogs in Foz do Iguaçu city. Hence, collaborative control measures should be designed and implemented by the public agencies and research institutions of Brazil, Argentina, and Paraguay to control the spread of visceral leishmaniasis.

https://doi.org/10.1590/s1984-29612019083 759 downloads

10.

Abundance and diversity of vectors (Diptera: Psychodidae) in an old transmission area of cutaneous leishmaniasis in the new world after Bolivia-Brazil gas pipeline construction
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Gonçalves, André Luiz

; Ennio, Luz

; Alcântara de Castro, Edilene

; Klisiowicz, Débora do Rocio

; Louise Gazda, Tatiana

; de Almeida Melo, André Luiz

; Thomaz-Soccol, Vanete

Memorias del Instituto de Investigaciones en Ciencias de la Salud

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aug 2019, Volume 17 N. 2 Pages 16 - 23

Abstract: En Es | Text: En Es | PDF: En | PDF: Es

ABSTRACT The objective of this study was to determine the abundance and diversity of the species and their presumed vectorial role in the transmission of Cutaneous Leishmaniasis (CL). Research on sand flies (Diptera: Psychodidae) was carried out in two municipalities in the Ribeira River Valley in the State of Paraná, southern Brazil. One, Adrianópolis, is an endemic area of CL and the other is a recent outbreak area (Cerro Azul). A total of 432 specimens were collected from residential, peri-domestic and wild environments. According to the data obtained, for each ecotope studied, the statistical analysis has shown that two lines of the Lutzomyia intermedia population coexist and that the quantity of specimens from L. intermedia s.l. is significantly different from L. intermedia s.s. in all environments (χ2=9.943; DF=2; p=0.07). The prevalence of L. intermedia (93.28%) in the ecotopes studied suggests that it is the main vector of leishmaniasis. The succession of vector species and their involvement in the epidemiological cycle of L. braziliensis is discussed in this article.

RESUMEN El objetivo de este trabajo fue determinar la abundancia y diversidad de las especies y su supuesto papel vectorial en la transmisión de Leishmaniasis Cutánea (LC). La investigación sobre flebótomos (Diptera: Psychodidae) se llevó a cabo en dos municipios en el Valle del Río Ribeira en el Estado de Paraná, sur de Brasil. Las regiones estudiadas fueron Adrianópolis, área endémica de LC, y Cerro Azul, que se reporta como zona de brote reciente. Se recolectaron un total de 432 especímenes de ambientes residenciales, peri-domésticos y silvestres. El análisis estadístico realizado, a partir de los datos obtenidos para cada ecotopo estudiado, mostró que coexistían dos linajes de la población de Lutzomyia intermedia y que la cantidad de especímenes de L. intermedia s.l. fue significativamente diferente de L. intermedia s.s. en todos los entornos (χ2=9,943; GL=2; p=0,07). La prevalencia de L. intermedia (93,28%) en los ecotopos estudiados sugiere que es el principal vector de la leishmaniasis. La sucesión de especies de vectores y su participación en el ciclo epidemiológico de L. braziliensis se discute en este artículo.

https://doi.org/10.18004/mem.iics/1812-9528/2019.017.02.16-023 520 downloads

11.

Epidemiological relevance of dogs for the prevention of Toxoplasma gondii, Neospora caninum and Leptospira spp.
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Pinto-Ferreira, Fernanda

; Pasquali, Aline Kuhn Sbruzzi

; Thomaz-Soccol, Vanete

; Mitsuka-Breganó, Regina

; Caldart, Eloiza Teles

; Leandro, André de Souza

; Chiyo, Luciana

; Pozzolo, Eliane Maria

; Cubas, Patrícia

; Giordano, Lucienne Garcia Pretto

; Petterle, Ricardo Rasmussen

; Navarro, Italmar Teodorico

Revista Brasileira de Parasitologia Veterinária

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jul 2019, Volume 28 N. 3 Pages 383 - 394

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo Este estudo avaliou a soroprevalência de Toxoplasma gondii, Neospora caninum e Leptospira spp. em cães de Foz do Iguaçu, Paraná, Brasil. O teste de imunofluorescência indireta foi utilizado para detectar anticorpos anti-T. gondii e anti-N. caninum. Ensaio imunoenzimático e soroaglutinação microscópica foram utilizados para pesquisa de anticorpos anti-T. gondii e anti-Leptospira spp., respectivamente. Os resultados obtidos foram: 67,02% (435/649) das amostras reativas para T. gondii e, 1,38% (9/649) para N. caninum, ambas com ausência de variáveis significativas estatisticamente. Para Leptospira spp. os resultados indicaram soroprevalência de 23,11% (153/649). A análise das variáveis sem distinção de sorovar mostraram associação para caraterísticas intrísecas como raça, idade, estado nutricional e categoria de cães. Para as variáveis extrínsecas, a região da cidade e ter acesso à rua mostraram associação estatística (p<0,05). Os sorovares mais prevalentes foram: Canicola com 59,47% (91/153); Bratislava 13,07% (20/153) e Butembo 15,68% (24/153). As variáveis que compõem o modelo de análise multivariada ajustada usando como desfecho Leptospira spp. foram: idade, raça, estado nutricional e área. Para o sorovar Canicola, as variáveis significantes (p<0,05) foram sexo, estado nutricional e área; para o sorovar Bratislava, as variáveis significantes (p<0,05) foram linfadenomegalia e presença de pulgas. Dos resultados obtidos, cães podem ser usados como sentinelas para infecção por T. gondii e Leptospira spp. na cidade de Foz do Iguaçu e em outras cidades com desfechos similares. Além disso, medidas preventivas devem ser tomadas pelas autoridades de saúde, pois são zoonoses e os seres humanos também estão em risco.

Abstract This study evaluated the seroprevalence of Toxoplasma gondii, Neospora caninum and Leptospira spp. in dogs from Foz do Iguaçu, Paraná, Brazil. Indirect immunofluorescent antibody test was used to detect antibodies anti-T.gondii and anti-N. caninum. Immunoenzymatic assay and microscopic serum agglutination were used for screening antibodies anti-T.gondii and anti-Leptospira spp., respectively. The results were: 67.02% of the samples reactive for T.gondii and 1.38% for N. caninum, both without statistically significant variables. For Leptospira spp. the results indicated seroprevalence of 23.11%. The analysis of the variables without distinction of serovar showed association for intrinsic characteristics as breed, age, nutritional status and dog category. The extrinsic variables as city region and access to the street presented association (p<0.05). The most prevalent serovars were: Canicola 59.47%; Bratislava 13.07% and Butembo 15.68%. Variables that make up the adjusted multiple analysis model using Leptospira spp. were: age, breed and nutritional status; serovar Canicola, sex, nutritional status and area (p<0.05); serovar Bratislava, lymphadenomegaly and presence of fleas (p<0.05). Given the results obtained, dogs can be used as sentinels for toxoplasmosis and leptospirosis in Foz do Iguaçu and other cities with similar outcomes. In addition, preventive measures should be taken by health authorities because they are zoonoses and humans are also at risk.

https://doi.org/10.1590/s1984-29612019043 1225 downloads

12.

Variables associated with the prevalence of anti-Leishmania spp. antibodies in dogs on the tri-border of Foz do Iguaçu, Paraná, Brazil
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Dias, Renata Cristina Ferreira

; Thomaz-Soccol, Vanete

; Pasquali, Aline Kuhn Sbruzzi

; Alban, Silvana Maria

; Fendrich, Ricardo Cancio

; Pozzolo, Eliane Maria

; Chiyo, Luciana

; Bisetto Júnior, Alceu

; Ferreira, Fernanda Pinto

; Desto, Stela Silva

; Freire, Roberta Lemos

; Mitsuka-Breganó, Regina

; Navarro, Italmar Teodorico

Revista Brasileira de Parasitologia Veterinária

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sep 2018, Volume 27 N. 3 Pages 338 - 347

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo O objetivo deste estudo foi investigar a ocorrência de anticorpos anti- Leishmania spp. em cães da cidade de Foz do Iguaçu, estado do Paraná, Brasil, fronteira com a Argentina e o Paraguai. Amostras de sangue de cães foram coletadas para realização dos seguintes testes sorológicos: teste rápido imunocromatográfico DPP®, ensaio imunoenzimático indireto (ELISA) e ensaio de imunofluorescência indireta (IFI). Em 2012, 285 cães foram analisados na fronteira com Argentina e, em 2013, amostras de soro de 396 cães na fronteira com o Paraguai. Utilizando ELISA para triagem e IFA para o teste confirmatório, os resultados mostraram uma prevalência de anticorpos de 1,8% (5/285) na fronteira da Argentina e 3,0% (12/396) na fronteira com o Paraguai. Ao usar o DPP® para triagem e ELISA como uma análise confirmatória, observou-se uma prevalência de cães sororreagentes de 2,5% (7/285) na fronteira com a Argentina e 5,1% (20/396) na fronteira com o Paraguai. A não coleta pública de lixo doméstico (p = 0,0004) mostrou-se associada à leishmaniose. Este estudo demonstra a presença de leishmaniose e sugere a emergência da leishmaniose visceral canina no estado do Paraná devido à ocorrência de cães sororreagentes na fronteira Argentina e Paraguai, que possuem características ambientais e geográficas que favorecem a disseminação do parasito.

Abstract The aim of this study was to investigate the occurrence of anti-Leishmania spp. antibodies in dogs from localities in the city of Foz do Iguaçu, Paraná state, Brazil, on the border with Argentina and Paraguay. Blood samples dogs were collected to perform the following serologic tests: immunochromatographic DPP® rapid test, indirect immunoenzymatic assay (ELISA) and indirect immunofluorescence assay (IFA). In 2012, 285 dogs were analyzed on Argentina border, and in 2013, serum samples from 396 dogs on the border of Paraguay were collected. Using ELISA for screening and IFA for the confirmatory test, the results showed that the antibody prevalence was 1.8% (5/285) on the border of Argentina and 3.0% (12/396) on Paraguay border. When using the DPP® for screening and ELISA as a confirmatory analysis, we observed a seroreagent prevalence in dogs of 2.5% (7/285) on Argentina border and 5.1% (20/396) on Paraguay border. The non-public collection of domestic waste (p= 0.0004) was shown to be associated with leishmaniasis. This study shows the presence of leishmaniasis and suggest the emergence of canine visceral leishmaniasis in state of Paraná due to the confirmed occurrence of seroreactive dogs on Argentina and Paraguay border, which has environmental and geographical characteristics that favor the spread of the parasite.

https://doi.org/10.1590/s1984-296120180055 1194 downloads

13.

New strategy to improve quality control of Montenegro skin test at the production level
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Guedes, Deborah Carbonera

; Minozzo, João Carlos

; Pasquali, Aline Kuhn Sbruzzi

; Faulds, Craig

; Soccol, Carlos Ricardo

; Thomaz-Soccol, Vanete

Revista da Sociedade Brasileira de Medicina Tropical

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dec 2017, Volume 50 N. 6 Pages 788 - 794

Abstract: En | Text: En | PDF: En

Abstract INTRODUCTION: The production of the Montenegro antigen for skin test poses difficulties regarding quality control. Here, we propose that certain animal models reproducing a similar immune response to humans may be used in the quality control of Montenegro antigen production. METHODS: Fifteen Cavia porcellus (guinea pigs) were immunized with Leishmania amazonensis or Leishmania braziliensis , and, after 30 days, they were skin tested with standard Montenegro antigen. To validate C. porcellus as an animal model for skin tests, eighteen Mesocricetus auratus (hamsters) were infected with L. amazonensis or L. braziliensis , and, after 45 days, they were skin tested with standard Montenegro antigen. RESULTS: Cavia porcellus immunized with L. amazonensis or L. braziliensis , and hamsters infected with the same species presented induration reactions when skin tested with standard Montenegro antigen 48-72h after the test. CONCLUSIONS: The comparison between immunization methods and immune response from the two animal species validated C. porcellus as a good model for Montenegro skin test, and the model showed strong potential as an in vivo model in the quality control of the production of Montenegro antigen.

https://doi.org/10.1590/0037-8682-0131-2017 1121 downloads

14.

Genetic variability of populations of Nyssomyia neivai in the Northern State of Paraná, Brazil
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Gasparotto, Jaqueline de Carvalho

; Costa-Ribeiro, Magda Clara Vieira da

; Thomaz-Soccol, Vanete

; Liebel, Sandra Mara Rodrigues da Silva

; Neitzke-Abreu, Herintha Coeto

; Reinhold-Castro, Kárin Rosi

; Cristovão, Edilson Colhera

; Teodoro, Ueslei

Revista do Instituto de Medicina Tropical de São Paulo

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2017, Volume 59 elocation e10

Abstract: En | Text: En | PDF: En

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.

https://doi.org/10.1590/s1678-9946201759010 1603 downloads

15.

Probability of occurrence of the Brazilian spotted fever in northeast of Paraná state, Brazil
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Otomura, Flávio Haragushiku

; Truppel, Jessé Henrique

; Moraes Filho, Jonas

; Labruna, Marcelo Bahia

; Rossoni, Diogo Francisco

; Massafera, Rubens

; Soccol, Vanete Thomaz

; Teodoro, Ueslei

Revista Brasileira de Parasitologia Veterinária

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dec 2016, Volume 25 N. 4 Pages 394 - 400

Abstract: Pt En | Text: Pt En | PDF: Pt | PDF: En

Resumo A febre maculosa brasileira (FMB) é uma zoonose fatal devido às dificuldades para diagnosticá-la e tratá-la precocemente. A ocorrência de casos de FMB no Estado do Paraná suscitou a investigação de áreas de risco desta rickettsiose nos municípios de Japira, Jaboti, Pinhalão e Tomazina, na mesorregião norte pioneiro do Paraná. Para determinar as áreas de risco foram analisadas amostras de soro de 592 cães e 230 equídeos submetidos à reação de imunofluorescência indireta para Rickettsia rickettsii e R. parkeri. Além disto, foram construídos mapas de probabilidade de risco pela técnica de krigagem indicatriz. Das amostras testadas 5,3% (43/822) continham anticorpos para pelo menos uma das duas rickettsias testadas. Os equídeos apresentaram uma positividade de 7,8% (18/230) e os cães de 4,2% (25/592). A análise geoestatística mostrou que a soropositividade média é de 5 a 6%. Embora as soropositividade médias de cães e equídeos constatadas tenham sido menores do que as relatadas em áreas endêmicas do território brasileiro, os componentes bióticos (agente etiológico, vetor e reservatórios) e ambientais da epidemiologia da FMB se fazem presentes nos municípios referidos.

Abstract Brazilian spotted fever (BSF) is a fatal zoonosis because of the difficulties in its early diagnosis and treatment. Occurrences of BSF in the northeast of the state of Paraná prompted investigation of areas at risk of this rickettsiosis in the municipalities of Japira, Jaboti, Pinhalão and Tomazina. To determine the areas at risk, 592 serum samples from dogs and 230 from equids were analyzed by means of the indirect immunofluorescence assay (IFA) for Rickettsia rickettsii and R. parkeri . In addition, risk probability maps were drawn up using the kriging indicator technique. Among the samples tested, 5.3% (43/822) indicated presence of antibodies reactive to at least one of the two Rickettsia species tested: 7.8% of the equids (18/230) and 4.2% of the dogs (25/592) were positive. Geostatistical analysis showed that the average seropositivity rate was 5 to 6%. Although the average seropositivity rates observed among these dogs and equids were lower than those reported from endemic areas of Brazil, the biotic components (etiological agent, vector and reservoirs) and environmental aspects of BSF epidemiology were present in these municipalities.

https://doi.org/10.1590/s1984-29612016060 1222 downloads

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