RESUMO
Leishmaniosis is a great public health problem affecting both humans and animals. The disease is caused by the protozoan Leishmania spp., which has a complex cycle involving a phlebotomine vector. The ELISA test (Enzyme-Linked Immunosorbent Assay) along with a chromatographic immunoassay was defined by the Brazil Health Ministry as the confirmatory screening protocol in 2011. Uruguaiana city is 630 km away from Porto Alegre, which makes it difficult to send samples and diagnose leishmaniasis, as well as receive quick results. In view of this, the present study evaluated an in-house indirect ELISA method compared to indirect immunofluorescence assay (IFA) and dual-path platform chromatographic immunoassay (DPP-BioManguinhos®) for the detection of an immune response to Leishmania spp. in canine species. The serological evaluation included 48 canines from the western border of Brazil (Uruguaiana and Barra do Quaraí city). Among the 48 canine samples tested, 18 were positive when using the ELISA technique, 19 were positive with IFA, and 17 were positive with rapid test DPP®. The ELISA technique showed a sensitivity/specificity of 83.3%/86.7% when compared to IFA and 100%/96.8% compared to DPP®. The present study showed a prevalence of 37.5%, demonstrating that the infection circulates in the studied population. It can be concluded that the ELISA technique was valuable for use in field conditions when performing screening tests in endemic areas.
A leishmaniose é um grande problema de saúde pública que afeta tanto humanos quanto animais. A doença é causada pelo protozoário Leishmania spp., que possui um ciclo complexo envolvendo um vetor flebotomíneo. O teste ELISA (Enzyme-Linked Immunosorbent Assay), juntamente com um imunoensaio cromatográfico, foi definido pelo Ministério da Saúde do Brasil como protocolo de triagem confirmatória em 2011. A cidade de Uruguaiana fica a 630 km de Porto Alegre, o que dificulta o envio de amostras e o diagnóstico da leishmaniose, além da demora para obter os resultados. Diante disso, o presente estudo teve como objetivo avaliar um método de ELISA indireto in-house comparado ao ensaio de imunofluorescência indireta (IFA) e imunoensaio cromatográfico de plataforma de dupla via (DPP-BioManguinhos®) para a detecção de uma resposta imune contra Leishmania spp. na espécie canina. A avaliação sorológica incluiu 48 caninos da fronteira oeste do Brasil (cidade de Uruguaiana e Barra do Quaraí). Das 48 amostras caninas testadas, 18 foram positivas na técnica de ELISA, 19 foram positivas com IFA e 17 foram positivas com o teste rápido DPP®. A técnica de ELISA apresentou sensibilidade/especificidade de 83,3%/86,7% quando comparada ao IFA e 100%/96,8% em relação ao DPP®. O presente estudo apresentou prevalência de 37,5%, demonstrando que a infecção circula na população estudada. Pode-se concluir que a técnica de ELISA foi valiosa para uso em condições de campo na realização de testes de triagem em áreas endêmicas.
Assuntos
Animais , Cães , Ensaio de Imunoadsorção Enzimática/veterinária , Leishmaniose/veterinária , Saúde Pública , Doenças do Cão , Leishmania/patogenicidadeRESUMO
ABSTRACT: Leishmaniosis is a great public health problem affecting both humans and animals. The disease is caused by the protozoan Leishmania spp., which has a complex cycle involving a phlebotomine vector. The ELISA test (Enzyme-Linked Immunosorbent Assay) along with a chromatographic immunoassay was defined by the Brazil Health Ministry as the confirmatory screening protocol in 2011. Uruguaiana city is 630 km away from Porto Alegre, which makes it difficult to send samples and diagnose leishmaniasis, as well as receive quick results. In view of this, the present study evaluated an in-house indirect ELISA method compared to indirect immunofluorescence assay (IFA) and dual-path platform chromatographic immunoassay (DPP-BioManguinhos®) for the detection of an immune response to Leishmania spp. in canine species. The serological evaluation included 48 canines from the western border of Brazil (Uruguaiana and Barra do Quaraí city). Among the 48 canine samples tested, 18 were positive when using the ELISA technique, 19 were positive with IFA, and 17 were positive with rapid test DPP®. The ELISA technique showed a sensitivity/specificity of 83.3%/86.7% when compared to IFA and 100%/96.8% compared to DPP®. The present study showed a prevalence of 37.5%, demonstrating that the infection circulates in the studied population. It can be concluded that the ELISA technique was valuable for use in field conditions when performing screening tests in endemic areas.
RESUMO: A leishmaniose é um grande problema de saúde pública que afeta tanto humanos quanto animais. A doença é causada pelo protozoário Leishmania spp., que possui um ciclo complexo envolvendo um vetor flebotomíneo. O teste ELISA (Enzyme-Linked Immunosorbent Assay), juntamente com um imunoensaio cromatográfico, foi definido pelo Ministério da Saúde do Brasil como protocolo de triagem confirmatória em 2011. A cidade de Uruguaiana fica a 630 km de Porto Alegre, o que dificulta o envio de amostras e o diagnóstico da leishmaniose, além da demora para obter os resultados. Diante disso, o presente estudo teve como objetivo avaliar um método de ELISA indireto in-house comparado ao ensaio de imunofluorescência indireta (IFA) e imunoensaio cromatográfico de plataforma de dupla via (DPP-BioManguinhos®) para a detecção de uma resposta imune contra Leishmania spp. na espécie canina. A avaliação sorológica incluiu 48 caninos da fronteira oeste do Brasil (cidade de Uruguaiana e Barra do Quaraí). Das 48 amostras caninas testadas, 18 foram positivas na técnica de ELISA, 19 foram positivas com IFA e 17 foram positivas com o teste rápido DPP®. A técnica de ELISA apresentou sensibilidade/especificidade de 83,3%/86,7% quando comparada ao IFA e 100%/96,8% em relação ao DPP®. O presente estudo apresentou prevalência de 37,5%, demonstrando que a infecção circula na população estudada. Pode-se concluir que a técnica de ELISA foi valiosa para uso em condições de campo na realização de testes de triagem em áreas endêmicas.
RESUMO
The aim of this study was to characterize Leishmania spp. from canine and feline samples using Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP). It was conducted in the southern region of Brazil, located at border crossings to Argentina and Uruguay. Samples were collected from 116 dogs (Canis lupus familiaris) and 89 cats (Felis catus). The PCR was performed to screen for an LT1 fragment from kinetoplast DNA (kDNA) target gene, and positive samples were subjected to a second PCR for an internal transcribed spacers (ITS1) region from ribosomal DNA (rDNA) target. RFLP was performed using the Haemophilus aegyptius (HAE III) restriction endonuclease (Fermentas ®). Positive samples by PCR ITS1 were sequenced and deposited in NCBI GenBank, and a phylogenetic analysis was developed. We found that 12.9% (15/116) of the samples from dogs were positive. All the 89 cat samples were negative. Positive samples were tested against Leishmania reference strains presenting different patterns in PCR-RFLP, and these samples showed bands denoting similarity to the standard species of Leishmania infantum, proven through sequencing and phylogenetic analysis. The RFLP technique, alone, was shown to be feasible for practical application and confirmation of the involved Leishmania spp.
Assuntos
Doenças do Gato , Doenças do Cão , Leishmania infantum , Leishmaniose Visceral , Animais , Animais Domésticos , Brasil , Doenças do Gato/diagnóstico , Doenças do Gato/epidemiologia , Doenças do Gato/parasitologia , Gatos , DNA de Cinetoplasto/genética , Doenças do Cão/diagnóstico , Doenças do Cão/parasitologia , Cães , Leishmania infantum/genética , Leishmaniose Visceral/veterinária , Filogenia , Reação em Cadeia da Polimerase/veterináriaRESUMO
The aim of this study was to characterize Leishmania spp. from canine and feline samples using Polymerase Chain Reaction (PCR)- Restriction Fragment Length Polymorphism (RFLP). It was conducted in the southern region of Brazil, located at border crossings to Argentina and Uruguay. Samples were collected from 116 dogs (Canis lupus familiaris) and 89 cats (Felis catus). The PCR was performed to screen for an LT1 fragment from kinetoplast DNA (kDNA) target gene, and positive samples were subjected to a second PCR for an internal transcribed spacers (ITS1) region from ribosomal DNA (rDNA) target. RFLP was performed using the Haemophilus aegyptius (HAE III) restriction endonuclease (Fermentas ®). Positive samples by PCR ITS1 were sequenced and deposited in NCBI GenBank, and a phylogenetic analysis was developed. We found that 12.9% (15/116) of the samples from dogs were positive. All the 89 cat samples were negative. Positive samples were tested against Leishmania reference strains presenting different patterns in PCR-RFLP, and these samples showed bands denoting similarity to the standard species of Leishmania infantum, proven through sequencing and phylogenetic analysis. The RFLP technique, alone, was shown to be feasible for practical application and confirmation of the involved Leishmania spp.(AU)
O objetivo deste trabalho foi caracterizar espécies de Leishmania em amostras de caninos e felinos, utilizando-se a reação em cadeia da polimerase (PCR)- polimorfismo de comprimento de fragmento de restrição (RFLP). O estudo foi realizado na região de fronteira no sul do Brasil, divisa com Argentina e Uruguai. Amostras foram coletadas de 116 cães (Canis lupus familiaris) e 89 gatos (Felis catus). A PCR foi realizada com o gene alvo do fragmento LT1 do DNA do cinetoplasto (kDNA) para triagem e, as amostras positivas foram submetidas a uma segunda PCR com alvo ITS1 no DNA ribossomal (rDNA). O RFLP foi realizado com a endonuclease de restrição Haemophilus aegyptius (HAE III) (Fermentas ®). As sequências positivas no PCR-ITS1 foram depositadas no NCBI GenBank, além disso, a análise filogenética foi realizada. Foram detectadas 12,9% (15/116) amostras positivas em cães. Das 89 amostras de gatos todas foram negativas. Cepas de referência de Leishmania, com padrões diferentes na PCR-RFLP, e amostras positivas apresentaram similaridade das bandas com as espécies padrão de Leishmania infantum, o que foi comprovado no sequenciamento e análise filogenética. A técnica de RFLP, sozinha, demonstrou viabilidade para a aplicação prática e a confirmação das espécies de Leishmania spp. envolvidas.(AU)
Assuntos
Animais , Leishmaniose/diagnóstico , Análise do Polimorfismo de Comprimento de Fragmentos Amplificados/instrumentação , Leishmania/classificação , Argentina , Uruguai , Áreas de Fronteira , BrasilRESUMO
Abstract Dexchlorpheniramine is a first-generation classical antihistamine, clinically used to treat allergies. The main objective of our study was to evaluate the effects of the dexchlorpheniramine reference standard (DCPA Ref. St) and a pharmaceutical formula on DNA in human peripheral blood mononuclear cells (PBMCs). We exposed PBMCs to five different concentrations (0.5, 2.5, 5, 10, and 50 ng/mL) of DCPA Ref. St DCPA Ref. St and pharmaceutical formula in order to evaluate their cytotoxic, genotoxic, and mutagenic potential. The results showed that both dexchlorpheniramine formulations did not affect PBMC viability and CD3+, CD4+, or CD8+ lymphocyte subpopulations. The DCPA Ref. St and pharmaceutical formula neither induced genotoxic or mutagenic effects nor numerical or structural chromosomal alterations in PBMCs after 24 hours of exposure.
Assuntos
Humanos , Leucócitos Mononucleares/metabolismo , Citotoxicidade Imunológica , Composição de Medicamentos , Genotoxicidade , Testes de Mutagenicidade , DNA/análise , Antagonistas dos Receptores Histamínicos/efeitos adversos , Hipersensibilidade/complicaçõesRESUMO
BACKGROUND: The Leishmaniases are on the top of the global list of tropical neglected diseases. The number of infected dogs in South America is estimated in millions and correlated to disease cases in humans, especially in Brazil. Equines may get infected too and can play a role in the epidemiological chain. Thus, the aim of the present study was to evaluate risk and protective factors of leishmaniasis in rural areas of the western border region of Rio Grande do Sul state, Brazil by Leishmania spp. protozoa molecular detection and serological evaluation (ELISA) in equine and canine blood samples. This work included nine farms around the city of Uruguaiana. Epidemiologic information regarding farm characteristics and biologic material collection of canine (22) and equine (91), totalizing 113 samples was collected. The polymerase chain reaction (PCR) technique was used to detect Leishmania spp. in biological samples. Variables related to the farm were collected and evaluated through descriptive analysis followed by chi-square test and a logistic regression analysis. RESULTS: Nineteen positive samples (19/113 - 16,81%) were detected, being 18 equines and 1 canine, in six of the nine farms included in the study. No animal showed clinical signs of the disease. According to the variables analyzed, when compared each characteristic separately, the presence of abundant vegetation and poor hygiene demonstrated to be risk factors to Leishmania infection in rural areas. The logistic regression showed that excellent general hygiene, proximity to the weir and trimmed grass were protective factors (p=0.038, p=0.001 and p=0.014, respectively). Having excellent hygiene represents a 70% lower chance of getting infected, keeping the grass cut protects the animal by more than 90% and the proximity of the weir represents a protective factor of 96%. CONCLUSIONS: The presence of Leishmania infection in the western border region of Rio Grande do Sul was 16,81% and it was influenced by farm characteristics. The role of the excellent general hygiene as a protective factor is extremely relevant in the leishmaniases prevention.
Assuntos
Doenças do Cão/parasitologia , Doenças dos Cavalos/parasitologia , Leishmaniose/veterinária , Criação de Animais Domésticos , Animais , Brasil/epidemiologia , Doenças do Cão/epidemiologia , Cães , Doenças dos Cavalos/epidemiologia , Cavalos , Higiene , Leishmaniose/epidemiologia , Fatores de Risco , População RuralRESUMO
Yacon is an Andean plant that has been used in folk medicine for its medicinal properties. The beneficial effects of this plant are possibly due to the high content of phenolic compounds present in its leaves and roots. This study evaluated the in vitro toxicity of the hydroalcoholic extract of leaves and roots from yacon (1, 10, 50, and 100 µg/mL) through cell viability tests, genotoxic and mutagenic activity in leukocytes culture cells; and cytotoxicity and apoptosis cell death (1, 10, 50, 100, and 500 µg/mL) in cell line originally established from the primary mouse embryonic fibroblast cells that were cultured by the designated protocol, so-called 3T3 protocol "3-day transfer, inoculum 3 × 105 cells" (3T3 cell line). No mutagenic and cytotoxic activities were observed in leukocyte cultures. Cytotoxic activity was evidenced in the highest concentrations of yacon leaf extract (50 and 100 µg/mL), whereas all concentrations tested with yacon leaf extract there was induction for apoptosis in the 3T3 cells. Genotoxic potential was observed only at higher doses of leaf (50 and 100 µg/mL) and root (100 µg/mL) extract. These results suggest that yacon leaf at high concentrations may present toxic potential showing concentration-dependent behavior; however, in vivo studies should be performed to validate these results.
Assuntos
Asteraceae , Extratos Vegetais , Animais , Sobrevivência Celular , Fibroblastos , Camundongos , Fenóis/toxicidade , Extratos Vegetais/toxicidade , Folhas de PlantaRESUMO
Leishmaniasis is a neglected zoonotic disease caused by a variety of pathogenic Leishmania species. In the New World, especially in Brazil, visceral leishmaniasis (VL) is caused by Leishmania infantum. The pathogen can infect several animal species including dogs, foxes, rodents, primates, felines, equines and humans. Dogs act as the primary domestic reservoirs. This study aimed to use polymerase chain reaction (PCR) for detecting Leishmania infection in horses living in a canine visceral leishmaniasis (CVL) endemic region. DNA samples from horse peripheral blood were used to perform PCR. Templates were amplified using primers for the kinetoplast DNA (kDNA) minicircles, which were able to detect different species of Leishmania. In addition, primers for internal transcribed spacer (ITS) of ribosomal DNA were used for detection of Trypanosomatidae sp. Amongst the 75 (39%) positive PCR samples from total 192 samples, 21 samples were positive for kDNA and 63 samples were positive for either ITS, ITS1, or ITS2 gene markers. The kDNA PCR and sequencing allowed the detection of L. infantum in horse blood samples. To our knowledge, this is the first report of equine infection with L. infantum in Southern Brazil. These results proved that L. infantum could also infect horses in addition to humans and dogs, as well as in European countries. This conclusion emphasizes the urgent need to follow up investigation of the infection in these animals.(AU)
A leishmaniose é uma doença zoonótica negligenciada, causada por uma diversidade de espécies patogênicas de Leishmania. No Novo Mundo, especialmente no Brasil, a leishmaniose visceral é causada pelo protozoário Leishmania infantum. A infecção acomete várias espécies animais (cães, raposas, roedores, primatas, felinos, equinos) e humanos e o cão é o principal reservatório doméstico. Este estudo teve como objetivo utilizar a reação em cadeia da polimerase (PCR) para detectar a infecção por Leishmania sp. em cavalos que vivem em uma região endêmica para leishmaniose visceral canina (LVC). Amostras de DNA do sangue periférico de equinos foram utilizadas para extração de DNA e PCR. As amplificações foram realizadas utilizando-se marcadores para a região do DNA do cinetoplasto (kDNA), que são capazes de detectar diferentes espécies de Leishmania sp. e a região dos espaçadores internos transcritos (ITS) do DNA ribossomal para detecção da família Trypanosomatidae. Dentre os 192 animais testados, 75 (39%) amostras foram positivas no PCR, 21 amostras foram positivas para PCR kDNA e 63 para os genes ITS, ITS1 e ITS2. O sequenciamento dos amplicons permitiu detectar L. infantum em amostras de sangue periférico de equinos. Este foi o primeiro relato de infecção por L. infantum no sul do Brasil na espécie equina. Os resultados demonstram que, assim como nos países europeus, a infecção foi detectada em equinos após ter sido identificada em seres humanos e cães, o que indica a urgência em acompanhar a investigação da infecção nessa espécie animal.(AU)
Assuntos
Animais , Reação em Cadeia da Polimerase/veterinária , Leishmania infantum , Cavalos , Leishmaniose Visceral/diagnósticoRESUMO
Leishmaniasis is a neglected zoonotic disease caused by a variety of pathogenic Leishmania species. In the New World, especially in Brazil, visceral leishmaniasis (VL) is caused by Leishmania infantum. The pathogen can infect several animal species including dogs, foxes, rodents, primates, felines, equines and humans. Dogs act as the primary domestic reservoirs. This study aimed to use polymerase chain reaction (PCR) for detecting Leishmania infection in horses living in a canine visceral leishmaniasis (CVL) endemic region. DNA samples from horse peripheral blood were used to perform PCR. Templates were amplified using primers for the kinetoplast DNA (kDNA) minicircles, which were able to detect different species of Leishmania. In addition, primers for internal transcribed spacer (ITS) of ribosomal DNA were used for detection of Trypanosomatidae sp. Amongst the 75 (39%) positive PCR samples from total 192 samples, 21 samples were positive for kDNA and 63 samples were positive for either ITS, ITS1, or ITS2 gene markers. The kDNA PCR and sequencing allowed the detection of L. infantum in horse blood samples. To our knowledge, this is the first report of equine infection with L. infantum in Southern Brazil. These results proved that L. infantum could also infect horses in addition to humans and dogs, as well as in European countries. This conclusion emphasizes the urgent need to follow up investigation of the infection in these animals.
A leishmaniose é uma doença zoonótica negligenciada, causada por uma diversidade de espécies patogênicas de Leishmania. No Novo Mundo, especialmente no Brasil, a leishmaniose visceral é causada pelo protozoário Leishmania infantum. A infecção acomete várias espécies animais (cães, raposas, roedores, primatas, felinos, equinos) e humanos e o cão é o principal reservatório doméstico. Este estudo teve como objetivo utilizar a reação em cadeia da polimerase (PCR) para detectar a infecção por Leishmania sp. em cavalos que vivem em uma região endêmica para leishmaniose visceral canina (LVC). Amostras de DNA do sangue periférico de equinos foram utilizadas para extração de DNA e PCR. As amplificações foram realizadas utilizando-se marcadores para a região do DNA do cinetoplasto (kDNA), que são capazes de detectar diferentes espécies de Leishmania sp. e a região dos espaçadores internos transcritos (ITS) do DNA ribossomal para detecção da família Trypanosomatidae. Dentre os 192 animais testados, 75 (39%) amostras foram positivas no PCR, 21 amostras foram positivas para PCR kDNA e 63 para os genes ITS, ITS1 e ITS2. O sequenciamento dos amplicons permitiu detectar L. infantum em amostras de sangue periférico de equinos. Este foi o primeiro relato de infecção por L. infantum no sul do Brasil na espécie equina. Os resultados demonstram que, assim como nos países europeus, a infecção foi detectada em equinos após ter sido identificada em seres humanos e cães, o que indica a urgência em acompanhar a investigação da infecção nessa espécie animal.
Assuntos
Animais , Cavalos , Leishmania infantum , Leishmaniose Visceral/diagnóstico , Reação em Cadeia da Polimerase/veterináriaRESUMO
Visceral leishmaniasis is an endemic zoonotic disease identified especially in developing territories. Brazil's northeast, southeast and midwest have been endemic for several years; currently, the infection is spreading to the south. Dogs are the main reservoirs; however, other mammal species have also been infected. Herein, we have identified the infecting Leishmania species in dogs and horses from the south of Brazil, a new outbreak of the infection. Blood samples were collected in the urban area of Uruguaiana city. DNA was extracted from peripheral blood, kinetoplast DNA (kDNA) and ribosomal DNA (rDNA) fragments were obtained by polymerase chain reaction (PCR) and sequenced. Out of 123 samples, 25 of them (14 dogs and 11 horses) were positive for Leishmania spp. Sequence alignment and phylogenetic analysis revealed that the kDNA in positive samples was similar to four species previously reported: L. infantum/L. chagasi, L. donovani, L. major. Despite kDNA minicircles regions are very useful due to high sensitivity to Leishmania spp. DNA detection, the sequence polymorphism among minicircles can be an obstacle to interspecific differentiation. Our results suggest that these strains are circulating in Brazil south region cross-border and indicate the susceptibility of new outbreak for visceral leishmaniasis infection in horses domiciled in endemic region for canine and human visceral leishmaniasis.
Assuntos
Reservatórios de Doenças/parasitologia , Doenças do Cão/epidemiologia , Doenças dos Cavalos/epidemiologia , Leishmania donovani/genética , Leishmaniose Visceral/veterinária , Zoonoses/epidemiologia , Animais , Brasil/epidemiologia , DNA de Cinetoplasto/genética , Doenças do Cão/parasitologia , Cães , Feminino , Doenças dos Cavalos/parasitologia , Cavalos , Humanos , Leishmania donovani/isolamento & purificação , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , Masculino , Mamíferos , Filogenia , Reação em Cadeia da Polimerase/veterinária , Alinhamento de Sequência/veterinária , Zoonoses/parasitologiaRESUMO
BACKGROUND: The γ-hexalactone is a flavoring agent for alcoholic beverages, teas, breads, dairy products, coffees, buttery products among others. It presents low molecular weight and exhibits sweet fruity aroma with nuances of nuts. As far as we know, both literature and government regulations have gaps regarding the safe use of the γ-hexalactone. In this context, the main objective of this work was to evaluate the effects of γ-hexalactone through in silico and in vitro approaches. METHODS: The in silico analysis was performed through four free online platforms (admetSAR, Osiris Property Explorer®, pkCSM platform and PreADMET) and consisted of comparative structural analysis with substances present in databases. The computational prediction was performed in the sense of complement and guide the in vitro tests. Regarding in vitro investigations, screening of cytotoxicity (assessed by cell proliferation and viability parameters) in lymphocytes exposed to γ-hexalactone for 72 h were carried out previously to determine non-cytotoxic concentrations. Following this screening, concentrations of 5.15, 0.515, and 0.0515 µM were selected for the study of the respective potentials: genotoxic (assessed by DNA comet assay), chromosomal mutation (analysis of micronucleus frequency) and immunomodulatory (cytokine quantification using ELISA immunoassay). The results of in vitro assays were compared by one-way analysis of variance (ANOVA), followed by Bonferroni's post hoc test, conducted by statistic software. RESULTS: The platform PreADMET pointed out that γ-hexalactone is potentially mutagenic and carcinogenic. The comet assay data corroborate with these results demonstrating that γ-hexalactone at 5.15 µM caused lymphocytes DNA damage. In relation to cytokine secretion, the results indicate that lymphocytes were activated by γ-hexalactone at non-cytotoxic concentrations, involving an increase in the IL-1 levels in all tested concentrations, ranging from approximately 56 to 93%. The γ-hexalactone only at 5.15 µM induced increase in the levels of IL-6 (~ 60%), TNF-α (~ 68%) and IFN-γ (~ 29%), but decreased IL-10 (~ 46%) in comparison with the negative control (p < 0.05). No change was observed in total lymphocytes or in cell viability at the concentrations tested. CONCLUSIONS: In summary, the γ-hexalactone demonstrated immunomodulatory and genotoxic effects at non-cytotoxic concentrations in healthy lymphocytes.
Assuntos
Citocinas/metabolismo , Dano ao DNA , Aromatizantes/toxicidade , Lactonas/toxicidade , Micronúcleos com Defeito Cromossômico/induzido quimicamente , Mutagênicos/toxicidade , Adulto , Proliferação de Células/efeitos dos fármacos , Sobrevivência Celular/efeitos dos fármacos , Células Cultivadas , Ensaio Cometa , Simulação por Computador , Relação Dose-Resposta a Droga , Humanos , Concentração Inibidora 50 , Leucócitos Mononucleares/efeitos dos fármacos , Leucócitos Mononucleares/metabolismo , Adulto JovemRESUMO
BACKGROUND: Leishmaniosis, zoonosis that produces significant public health impacts, is caused by Leishmania infantum. Canines are the main domestic reservoir and, besides humans, other species of mammals could be infected when living in endemic areas. In this study, we detected equine Leishmania infantum infections in a canine visceral leishmaniosis transmission area and evaluated the clinical, haematological, biochemical and oxidative stress disorders. This study was conducted in Uruguaiana, Rio Grande do Sul, south of Brazil. Peripheral blood samples were collected from 124 animals (98 horses and 26 dogs) of both genders and several breeds after they underwent general and dermatologic examinations. RESULTS: Twenty five Leishmania infantum infected animals (20.16%), 14 horses and 11 dogs were detected by PCR (Polymerase Chain Reaction) amplification of kinetoplast DNA regions with 96% homology to Leishmania infantum (GenBank Accession No. L 19877.1). The clinical and haematological alterations of infected equines were skin lesions, nodules, lymphadenopathy, decreased levels in red blood cells and haematocrit (p < 0.05) and increase in urea serum concentration (p < 0.05), while CVL presented a decrease in red blood cells counts (p < 0.05), increase in lymphocytes (p < 0.05), and decrease in neutrophil-lymphocyte ratio (p < 0.05). Oxidative stress markers of plasma protein carbonyl and plasma lipid peroxidation were not statistically significant (p > 0.05) in both species. CONCLUSIONS: To our knowledge, this has been the first leishmaniosis equine survey performed in south of Brazil, caused by Leishmania infantum that were able to initially identify haematological and biochemical changes in the species, even in asymptomatic animals. We present evidence supporting those findings of haematological and biochemical changes could be related to infection. Surprisingly, the clinical manifestations of equine infection were similar to those found in canine visceral leishmaniosis. The equine population could be play an important role in the cycle of leishmaniosis in south Brazil and consequently indicates a great risk of public health. This evaluation of infected animals is important to establish the clinical and laboratory parameters involved in the disease progression.
Assuntos
Doenças do Cão/parasitologia , Doenças dos Cavalos/parasitologia , Leishmania infantum/isolamento & purificação , Leishmaniose Visceral/veterinária , Animais , Brasil/epidemiologia , DNA de Cinetoplasto/genética , Doenças do Cão/sangue , Doenças do Cão/epidemiologia , Cães , Doenças dos Cavalos/sangue , Doenças dos Cavalos/epidemiologia , Cavalos , Leishmania infantum/genética , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/parasitologia , ZoonosesRESUMO
Fluazuron is one of the newest veterinary antitick medicines. Belonging to the benzoylphenylureas group, its mechanism of action acts by the interference of the formation of the chitin of the tick, which is responsible for the hardening of its exoskeletons. In addition to taking care of the health of the animal so that it receives the medication in the doses and the correct form, it is important to analyze the safety of the operator. Reduced resistance to infectious disease was a well-documented consequence of primary and acquired immunodeficiencies, but a novel finding following xenobiotic exposure. The awareness of the consequences of altered immune function is the most likely outcome of inadvertent exposure. The human health implications of studies in which chemical exposure reduced resistance to infection drove an early focus on immunosuppression within the toxicology community. The main objective is to perform the evaluation by computational platforms and in cell culture, searching for data that can serve as a foundation for a better understanding of the toxic effects involved with the accidental contamination of Fluazuron and, thus, to assist the medical community and users to understand the risks inherent in its use. As far as we can determine in the literature, our work has unmistakably demonstrated that the Fluazuron can cause genotoxicity by probable chromatin rearrangement and immunodepleting by specific reduction of the CD8 T lymphocyte subpopulation, mediated by the decrease in gamma interferon production. Although the use of Fluazuron is a necessity for tick control and for cattle management, we must bear in mind that the imminent risks to its application exist. Careless use can damage the immune system which in turn carries a gigantic hazard by opening a door to diseases and pathogens and leaving us defenseless.
RESUMO
This study evaluated the Limnoperna fortunei (golden mussel) as a bioindicator of cytotoxicity and genotoxicity in aquatic environments contaminated by heavy metals. Five groups of 50 subjects each were exposed to different concentration of mercuric chloride (HgCl2) (0.001â¯mg/L, group I; 0.005â¯mg/L, group II; 0.01â¯mg/L, group II; 0.02â¯mg/L, group IV; and 0.1â¯mg/L, group V). The control group for both chronic and acute treatment did not receive HgCl2. For chronic exposure, the respective groups were placed in aquaria with water contaminated with the above concentrations of HgCl2. For acute exposure, the different concentrations of HgCl2 were injected into the posterior adductor muscle of the individuals belonging to the aforementioned groups. The biological matrix used in the tests was the whole body muscle. Tests (cell viability assay, alkaline comet test; enumeration of micronuclei and necrotic cells, quantification of Hg content in tissues and water, and histopathological analysis of tissues), were carried out on the 7th, 15th, and 30th treatment days or 2â¯h after injection. Our results demonstrated that L. fortunei showed cell damage in both chronic and acute exposure groups. Significant DNA damage was observed at both the 15th (0.1â¯mg/L) and 30th (0.01-0.1â¯mg/L) days of chronic exposure. However, in acute treatment all concentrations induced DNA breaks. The presence of necrosis increased at all concentrations tested for both acute and chronic exposure. Tissue mercury retention on the 15th day was higher than on the 30th day of exposure, while in the same period, there was a decrease in the mercury content of aquarium water. Taking the data together, it is concluded that L. fortunei as a possible bioindicator of the quality of aquatic environments.
Assuntos
Monitoramento Ambiental , Mercúrio/toxicidade , Mytilidae/fisiologia , Poluentes Químicos da Água/toxicidade , Animais , Dano ao DNA , Biomarcadores AmbientaisRESUMO
The present study aimed to evaluate factors associated with canine visceral leishmaniasis (CVL) in areas with higher seroprevalence of it in Uruguaiana city, Rio Grande do Sul state, Brazil, during the year of 2012. A semi-structured questionnaire with socioeconomic and environmental questions was applied in Leishmania endemic regions of the city. The survey data were analyzed by multivariate statistics and the associations between them were presented as odds ratio within a 95% confidence limit. The relationships between the studied variables showed no statistically significant difference between the dwellings with positive and negative dogs. However, when analyzing the odds ratio, the presence of green areas larger than 10m² turned the house into an area of potential risk for CVL (OR= 2.53). There was no difference among the socioeconomic variables education and income, though the groups with lower education and income showed a higher seroprevalence of CVL. The city of Uruguaiana is already located within the CVL broadcasting area and is taken as an endemic region of the disease, with seropositive dogs in all neighborhoods. Thus, the determinants for an increased Leishmania sp. infection could not be indicated, even so, the maintenance of green areas around the households represented a risk factor for being a potential vector shelter.
O objetivo da presente pesquisa foi avaliar potenciais fatores de risco associados à ocorrência de leishmaniose visceral canina (LVC) em áreas de maior prevalência no município de Uruguaiana/RS, no ano de 2012. Foram aplicados questionários semiestruturados com questões socioeconômicas e ambientais em bairros endêmicos para LVC no município de Uruguaiana. Os dados foram analisados por meio de análise multivariada e as associações entre as variáveis foram expressas em odds ratio com intervalos de confiança de 95%. As relações entre as variáveis estudadas não apresentaram diferença estatística significativa quando se comparou residências com cães positivos àquelas com cães negativos. Entretanto, a presença de área verde maior que 10m² apresentou-se como um potencial fator de risco (OR = 2,53) para LVC. Não houve diferença significativa entre as variáveis socioeconômicas, escolaridade e renda embora, nos grupos de menor escolaridade e de menor renda familiar, tenha havido maior soro prevalência para LVC. O município de Uruguaiana-RS já pertence à área de transmissão de LVC tornando-se uma região endêmica da doença, com presença de cães soropositivos em todos os bairros. Desta forma, não foi possível apontar os determinantes para o aumento da infecção por Leishmania sp, embora a manutenção de áreas verdes peridomiciliares tenha representado um fator de risco devido à conservação do vetor nesses locais.
Assuntos
Animais , Doenças Negligenciadas/veterinária , Fatores de Risco , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/etiologia , Leishmaniose Visceral/veterinária , Brasil , Fatores SocioeconômicosRESUMO
The present study aimed to evaluate factors associated with canine visceral leishmaniasis (CVL) in areas with higher seroprevalence of it in Uruguaiana city, Rio Grande do Sul state, Brazil, during the year of 2012. A semi-structured questionnaire with socioeconomic and environmental questions was applied in Leishmania endemic regions of the city. The survey data were analyzed by multivariate statistics and the associations between them were presented as odds ratio within a 95% confidence limit. The relationships between the studied variables showed no statistically significant difference between the dwellings with positive and negative dogs. However, when analyzing the odds ratio, the presence of green areas larger than 10m² turned the house into an area of potential risk for CVL (OR= 2.53). There was no difference among the socioeconomic variables education and income, though the groups with lower education and income showed a higher seroprevalence of CVL. The city of Uruguaiana is already located within the CVL broadcasting area and is taken as an endemic region of the disease, with seropositive dogs in all neighborhoods. Thus, the determinants for an increased Leishmania sp. infection could not be indicated, even so, the maintenance of green areas around the households represented a risk factor for being a potential vector shelter.(AU)
O objetivo da presente pesquisa foi avaliar potenciais fatores de risco associados à ocorrência de leishmaniose visceral canina (LVC) em áreas de maior prevalência no município de Uruguaiana/RS, no ano de 2012. Foram aplicados questionários semiestruturados com questões socioeconômicas e ambientais em bairros endêmicos para LVC no município de Uruguaiana. Os dados foram analisados por meio de análise multivariada e as associações entre as variáveis foram expressas em odds ratio com intervalos de confiança de 95%. As relações entre as variáveis estudadas não apresentaram diferença estatística significativa quando se comparou residências com cães positivos àquelas com cães negativos. Entretanto, a presença de área verde maior que 10m² apresentou-se como um potencial fator de risco (OR = 2,53) para LVC. Não houve diferença significativa entre as variáveis socioeconômicas, escolaridade e renda embora, nos grupos de menor escolaridade e de menor renda familiar, tenha havido maior soro prevalência para LVC. O município de Uruguaiana-RS já pertence à área de transmissão de LVC tornando-se uma região endêmica da doença, com presença de cães soropositivos em todos os bairros. Desta forma, não foi possível apontar os determinantes para o aumento da infecção por Leishmania sp, embora a manutenção de áreas verdes peridomiciliares tenha representado um fator de risco devido à conservação do vetor nesses locais.(AU)
Assuntos
Animais , Leishmaniose Visceral/veterinária , Leishmaniose Visceral/epidemiologia , Leishmaniose Visceral/etiologia , Doenças Negligenciadas/veterinária , Fatores de Risco , Fatores Socioeconômicos , BrasilRESUMO
ABSTRACT Ocimum is one of the most important genera of the Lamiaceae family. Several studies about basil and its popular use reveal many characteristics of the herb, including its use as antioxidant, anti-aging, anti-inflammatory, anti-carcinogenic, anti-microbial, and cardiovascular agents, among others. In this paper, we evaluated genotoxic, oxidative, and anti-inflammatory parameters from the extract of Ocimum basilicum in different concentrations, using human leukocytes cultures exposed to challenging agents. Our results confirm that the O. basilicum extract acts as an antioxidant and effectively reverts or subjugates the effects of high oxidizing agents such as hydrogen peroxide. These actions are attributed to its composition, which is rich in polyphenols and flavonoids as well as compounds such as rosmarinic acid, all of which have well-known antioxidant activity. We also show that our basil extract presents anti-inflammatory properties, the mechanism of which is a composed interaction between the inhibition of pro-inflammatory mediator and the stimulation of anti-inflammatory cytokines. Although pharmacodynamics studies are necessary to evaluate the activities in vivo, our results demonstrated that basil could act as an antioxidant and anti-inflammatory and a possible alternative for medicinal treatment.
Assuntos
Extratos Vegetais/análise , Ocimum basilicum/classificação , Anti-Inflamatórios/farmacologia , Plantas Medicinais/metabolismo , Ocimum basilicum/efeitos adversos , Leucócitos/classificação , Antioxidantes/uso terapêuticoRESUMO
Euphorbia tirucalli (L.), commonly known as aveloz, has been indiscriminately used in popular medicine to treat various illnesses. However, some components can have devastating consequences. Injury to a cell's genetic material can cause mutations, cancer, and cell death. Our main goal in this work was to evaluate the genotoxic and cytotoxic effects of E. tirucalli extract on human leukocytes. For this purpose, we performed a phytochemical analysis to evaluate the plant's components. In the second step, we treated cultured human leukocytes with different concentrations of the dry extract of the plant and then evaluated the oxidative and genotoxic profiles of these leukocytes. We found that at 1% and 10% concentrations, the aveloz extract acted as a genotoxic agent that could damage DNA and increase oxidative damage. We conclude that despite its popular use, aveloz can act as a genotoxic agent, especially when it contains phorbol ester. Aveloz's indiscriminate use might actually promote tumors and therefore carry a considerable genetic risk for its users.
Assuntos
Dano ao DNA/efeitos dos fármacos , Euphorbia/química , Leucócitos/efeitos dos fármacos , Extratos Vegetais/toxicidade , Células Cultivadas , Cromatografia Líquida de Alta Pressão , Relação Dose-Resposta a Droga , Humanos , Testes de MutagenicidadeRESUMO
AIM: To determine the plasmatic iron content and evaluate the oxidative stress (OS) markers in subjects receiving blood therapy. METHODS: Thirty-nine individuals with unspecified anemia receiving blood transfusions and 15 healthy subjects were included in the study. Anemic subjects were divided into three subgrouP: (1) those that received up to five blood transfusions (n = 14); (2) those that received from five to ten transfusions (n = 11); and (3) those that received more than ten transfusions (n = 14). Blood samples were collected by venous arm puncture and stored in tubes containing heparin. The plasma and cells were separated by centrifugation and subsequently used for analyses. Statistical analyses were performed using Kruskal-Wallis analysis of variance followed by Dunn's multiple comparison tests when appropriate. RESULTS: The eletrophoretic hemoglobin profiles of the subjects included in this study indicated that no patients presented with hemoglobinopathy. Labile plasmatic iron, ferritin, protein carbonyl, thiobarbituric acid-reactive substances (TBARS) and dichlorofluorescein diacetate oxidation were significantly higher (P < 0.05), whereas total thiol levels were significantly lower (P < 0.05) in transfused subjects compared to controls. Additionally, the activity of catalase, superoxide dismutase and glutathione peroxidase were significantly lower in the transfused subjects (P < 0.05). Antioxidant enzyme activities and total thiol levels were positively correlated (P < 0.05), and negatively correlated with the levels of protein carbonyl and TBARS (P < 0.05). In contrast, protein carbonyl and TBARS were positively correlated (P < 0.05). Altogether, these data confirm the involvement of OS in patients following therapy with repeated blood transfusions. CONCLUSION: Our data reveal that changes in OS markers are correlated with levels of labile plasmatic iron and ferritin and the number of transfusions.