Chemical composition of various plant extracts and their in vitro efficacy in control of Fasciola hepatica eggs / Composição química de vários extratos vegetais e sua eficácia in vitro no controle de ovos de Fasciola hepática

Fasciolosis has been diagnosed in cattle, goats, sheep and horses in southern and southeastern Brazil. Effective alternative treatments are the targets of study. One promising alternative is the use of plant extracts. The aim of this study was to perform phytochemical analysis of extracts of Eugenia uniflora L., Harpagophytum procumbens, Psidium guajava L. and Stryphnodendron adstringens, and to evaluate the in vitro efficacy of these extracts on ovicidal activity in Fasciola hepatica. Plant extracts were analyzed for phytochemical properties. F. hepatica eggs were collected directly from the gallbladders of animals diagnosed as positive for fasciolosis on post mortem examination. One hundred eggs were incubated with 3 ml of each extract at concentrations of 0.10%, 0.25% and 0.50%, albendazole 0.50% (positive control) or tap water (negative control). To determine anti larval efficacy of each plant extract, hatched eggs were counted and the averages were used. Phytochemical analysis revealed the presence of phenolic compounds, tannins and terpenes in most extracts. E. uniflora L. extract was 100% effective at 0.10%, H. procumbens was effective at 0.25% and P. guajava L. and S. adstringens extracts were 100% effective at all concentrations tested. Taken together, the data suggested that ovicidal activity in F. hepatica is due to the presence of these bioactive compounds.(AU)

A fasciolose tem sido diagnosticada em bovinos, caprinos, ovinos e equinos no sul e sudeste do Brasil, sendo que tratamentos alternativos mais eficazes são alvos de estudo. Umas das alternativas promissoras é o uso de extratos vegetais no controle dessa e outras enfermidades. O objetivo deste estudo foi realizar a análise fitoquímica dos extratos de Eugenia uniflora L., Harpagophytum procumbens, Psidium guajava L. e Stryphnodendron adstringens, além de avaliar a eficácia in vitro desses extratos na atividade ovicida em Fasciola hepatica. Os extratos vegetais foram obtidos e analisados para determinação fitoquímica. Ovos de F. hepatica foram coletados diretamente das vesículas biliares de animais diagnosticados como positivos para fasciolose no exame post mortem. Cem ovos foram incubados com três mililitros de cada extrato nas concentrações de 0,10%, 0,25% e 0,50%, de albendazol a 0,50% (controle positivo) e água de torneira (controle negativo). Para determinar a eficácia de cada extrato vegetal os ovos eclodidos foram contados, e a média utilizada para os cálculos de eficácia. A análise fitoquímica revelou a presença de compostos fenólicos, taninos e terpenos na maioria dos extratos. O extrato de E. uniflora L. apresentou eficácia de 100% na concentração de 0,10%, o de H. procumbens a 0,25% e os extratos de P. guajava L. e S. adstringens apresentaram 100% de eficácia em todas as concentrações testadas. Assim, sugere-se que a atividade ovicida em F. hepatica seja devido à presença desses compostos bioativos.(AU)

Morphometric and vascular analyses and MMP-2 expression in bladders of animals with bovine enzootic haematuria

Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridiumaquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovinepapillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognosticmarkers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a varietyof cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantifythe expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigert’s resorcin-fuchsin staining methods. Themorphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesionswere differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation,ectasia, dilation and thickening and the material stained using the Weigert’s resorcin-fuchsin method was used to evaluatesclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia,haemorrhage, cystitis cystica and Brunn’s nests and the neoplastic lesions were haemangioma, myxoma, transitional cellcarcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differencesbetween the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed...

Morphometric and vascular analyses and MMP-2 expression in bladders of animals with bovine enzootic haematuria

Background: Bovine enzootic haematuria is a chronic, non-infectious disease caused by the ingestion of bracken Pteridiumaquilinum that contain ptaquiloside, substance responsible for the development of neoplasms in combination with bovinepapillomavirus infection. Studies examining metalloproteinases have shown that these enzymes may be useful as prognosticmarkers and many studies have correlated the intratumoural microvessel density and incidence of metastasis in a varietyof cancers, including bladder cancer. This study aimed to evaluate the urothelial morphology and vasculature and quantifythe expression of the enzyme metalloproteinase-2 in the bladder lesions of bovine with enzootic haematuria.Materials, Methods & Results: Forty bladders with macroscopic lesions were subjected to routine histological processing. The material was stained using the Hematoxylin-Eosin (HE) and Weigerts resorcin-fuchsin staining methods. Themorphometric analysis included the measurement of the total thickness of the urothelium. Microscopically, the lesionswere differentiated as neoplastic or non-neoplastic, and the vascular changes were characterised as vascular proliferation,ectasia, dilation and thickening and the material stained using the Weigerts resorcin-fuchsin method was used to evaluatesclerosis and the vascular fibroelastosis. The metalloproteinase expression was evaluated using an anti-metalloproteinase-2antibody. The main non-neoplastic lesions found in the urothelium included clear cell metaplasia, dysplasia, hyperplasia,haemorrhage, cystitis cystica and Brunns nests and the neoplastic lesions were haemangioma, myxoma, transitional cellcarcinoma, adenocarcinoma, in situ carcinoma and haemangiosarcoma. The urothelium measurements revealed differencesbetween the epithelial thicknesses of bladders with and without neoplasms. The bladder vasculature evaluation revealed...(AU)

Microscopic alterations in Fasciola hepatica from sheep treated with albendazole / Alterações microscópicas em Fasciola hepatica de ovelhas tratadas com albendazol

Currently, albendazole is one of the most commonly used drugs because of its affordability. The objective was to evaluate the histopathology of Fasciola hepatica specimens. For this, the efficacy test was performed on sheep treated with albendazole at the dose recommended for F. hepatica, in which the helminths recovered at necropsy were counted and separated for histology. Spermatogenic cells from parasites recovered from treated and control sheep were examined by microscopy. The fecal egg-count reduction test revealed 97.06% efficacy of albendazole in the treatment of F. hepatica. Changes in testicular tubule cells started 48 hours after treatment and became evident within 72 hours, at which point it became difficult to identify cell types. Primary and secondary spermatogonia became increasingly rare and intercellular vacuolization was more evident. Signs of apoptosis, with pycnotic nuclei and evidence of keriorrexia were observed at all times. Cell debris was identified 96 hours after treatment. The results indicated that parasitic spermatogenesis was severely affected by albendazole and demonstrated the importance of the use of histopathology for the diagnosis of therapeutic efficacy in field strains.(AU)

Na atualidade, o albendazol é uma das drogas mais usadas devido à sua acessibilidade econômica. O objetivo foi avaliar a histopatologia dos espécimes de Fasciola hepatica. Para isso, foi realizado o teste de eficácia em ovinos tratados com albendazol na dose recomendada para Fasciola hepatica, no qual os helmintos recuperados em necropsia foram contabilizados e separados para histologia. As células espermatogênicas de parasitas recuperados de ovinos tratados e controle foram examinadas por microscopia. O teste de redução de ovos por grama de fezes revelou 97,06% de eficácia do albendazol no tratamento de F. hepatica. As alterações nas células dos túbulos testiculares iniciaram-se 48 horas após o tratamento e tornaram-se evidentes em 72 horas, altura em que tornou-se difícil identificar os tipos de células. As espermatogônias primárias e secundárias tornaram-se cada vez mais raras e a vacuolização intercelular foi mais evidente. Sinais de apoptose, com núcleos picnóticos e evidência de cariorrexia foram observados em todos os momentos. Os detritos celulares foram identificados 96 horas após o tratamento. Os resultados indicaram que a espermatogênese parasitária foi severamente afetada pelo albendazol e demonstrou a importância do uso da histopatologia para o diagnóstico de eficácia terapêutica em cepas de campo.(AU)

Microscopic alterations in Fasciola hepatica from sheep treated with albendazole / Alterações microscópicas em Fasciola hepatica de ovelhas tratadas com albendazol

Currently, albendazole is one of the most commonly used drugs because of its affordability. The objective was to evaluate the histopathology of Fasciola hepatica specimens. For this, the efficacy test was performed on sheep treated with albendazole at the dose recommended for F. hepatica, in which the helminths recovered at necropsy were counted and separated for histology. Spermatogenic cells from parasites recovered from treated and control sheep were examined by microscopy. The fecal egg-count reduction test revealed 97.06% efficacy of albendazole in the treatment of F. hepatica. Changes in testicular tubule cells started 48 hours after treatment and became evident within 72 hours, at which point it became difficult to identify cell types. Primary and secondary spermatogonia became increasingly rare and intercellular vacuolization was more evident. Signs of apoptosis, with pycnotic nuclei and evidence of keriorrexia were observed at all times. Cell debris was identified 96 hours after treatment. The results indicated that parasitic spermatogenesis was severely affected by albendazole and demonstrated the importance of the use of histopathology for the diagnosis of therapeutic efficacy in field strains.(AU)

Na atualidade, o albendazol é uma das drogas mais usadas devido à sua acessibilidade econômica. O objetivo foi avaliar a histopatologia dos espécimes de Fasciola hepatica. Para isso, foi realizado o teste de eficácia em ovinos tratados com albendazol na dose recomendada para Fasciola hepatica, no qual os helmintos recuperados em necropsia foram contabilizados e separados para histologia. As células espermatogênicas de parasitas recuperados de ovinos tratados e controle foram examinadas por microscopia. O teste de redução de ovos por grama de fezes revelou 97,06% de eficácia do albendazol no tratamento de F. hepatica. As alterações nas células dos túbulos testiculares iniciaram-se 48 horas após o tratamento e tornaram-se evidentes em 72 horas, altura em que tornou-se difícil identificar os tipos de células. As espermatogônias primárias e secundárias tornaram-se cada vez mais raras e a vacuolização intercelular foi mais evidente. Sinais de apoptose, com núcleos picnóticos e evidência de cariorrexia foram observados em todos os momentos. Os detritos celulares foram identificados 96 horas após o tratamento. Os resultados indicaram que a espermatogênese parasitária foi severamente afetada pelo albendazol e demonstrou a importância do uso da histopatologia para o diagnóstico de eficácia terapêutica em cepas de campo.(AU)

Chenopodium ambrosioides L. essential oil and ethanol extract on control of canine Ancylostoma spp. / Extrato etanólico e óleo essencial de Chenopodium ambrosioides L. no controle de Ancylostoma spp. de cães

The use of Chenopodium ambrosioides L. has shown to be promising in the management of gastrointestinal nematodes. The objective of this study was to quantitate the yield and characterize the chemical composition of the essential oil of C. ambrosioides, as well as to evaluate the in vitro effect of the ethanolic extract and the essential oil in L3 of Ancylostoma spp. and the in vivo effect(s) of the essential oil in dogs. The effects of the ethanol extract and essential oil on Ancylostoma spp. were evaluated in vitro by exposing larvae to the extract at concentrations ranging from 0.005 g mL-1 to 0.2 g mL-1 and to essential oil at concentrations of 50, 100, and 150 ?L mL-1. For the in vivo test, 26 healthy dogs, naturally infected by Ancylostoma spp., were divided into three groups: F1 - cookies were administered without active principle; F2 - herbal cookies containing 37.5 ?L g-1 essential oil of C. ambrosioides L.; F3 - cookies plus a commercial formulation containing febantel, pyrantel, praziquantel, and ivermectin. Complete blood counts and serum biochemistry for AST, ALT, AF, urea, creatinine, total protein, and albumin were performed. The yield of the essential oil was 0.3% m v-1, and its major components included ?-terpinene (1.24%), p-cymene (4.83%), and ascaridol Z (87%) and E (5.04%) isomers. The concentrations of C. ambrosioides L. ethanol extract used were ineffectiveagainst Ancylostoma spp. larvae. The essential oil at a concentration of 150 μL mL-1 was effectiveagainst L3 larvae. In the in vivo study in dogs, the herbal cookies containing C. ambrosioides L essentialoil reduced the number of eggs per gram of feces.(AU)

O uso de Chenopodium ambrosioides L. tem se mostrado promissor no controle de nematoides gastrintestinais. Objetivou-se verificar o rendimento e a composição química do óleo essencial de C. ambrosioides, assim como avaliar o efeito in vitro do extrato etanólico e do óleo essencial em L3 de Ancylostoma spp., e o efeito in vivo do óleo essencial para cães. Para a atividade larvicida as larvas de Ancylostoma spp. foram colocadas na presença do extrato etanólico nas concentrações de 0,005 g mL-1, 0,01 g mL-1, 0,02 g mL-1, 0,03 g mL-1, 0,04 g mL-1; 0,05 g mL-1, 0,06 g mL-1, 0,08 g mL-1, 0,1 g mL-1, 0,12 g mL-1 e 0,2 g mL-1, do óleo essencial nas concentrações de 50 µL mL-1, 100 µL mL-1 e 150 µL mL-1. Para o teste in vivo utilizaram-se 26 cães, hígidos, infectados naturalmente por Ancylostoma spp., divididos em três grupos: F1 - foram administrados biscoitos sem princípio ativo; F2 - biscoitos fitoterápicos com óleo essencial de C. ambrosioides L na concentração de 37,5 µL g-1; F3 biscoitos acrescido de formulação comercial a base de febantel, pirantel, praziquantel e ivermectina. Foi realizado hemograma completo e bioquímica sérica para AST, ALT, FA, ureia, creatinina, proteína total e albumina. Verificou-se o rendimento de 0,3% m v-1 do óleo essencial e na sua composição química a presença de ?-terpineno (1,24 %), o p-cimeno (4,83 %) e ascaridol, nos seus isômeros Z (87%) e E (5,04 %), como componentemajoritário. O extrato etanólico de C. ambrosioides L., nas concentrações utilizadas, apresentouinsuficiente atividade para o efeito larvicida. O óleo essencial na concentração de 150 µL mL-1 foi eficazcontra L3. A formulação de biscoito fitoterápico com óleo essencial C. ambrosioides L reduziu o númerode ovos por grama de fezes dos cães.(AU)

PCR Amplification of DNA Sequence of Bovine Papillomavirus Type 2 in Urinary Bladder of Cattle with Enzootic Hematuria in Espirito Santo, Brazil

The goal of this study was to investigate the presence of bovine papillomavirus type 2 (BPV-2) genomic DNA using polymerase chain reaction (PCR) assays, in the urinary bladder of cattle with bovine enzootic hematuria (BEH), in the neoplastic and non-neoplastic lesions in Espírito Santo, Brazil. Thus, 46 bladders with gross BEH lesions were collected from adult cattle at slaughterhouses. The samples were divided into two portions: one for use in histopathological analysis and the other one for PCR. Microscopic analysis showed that 56.52% of the bladders presented neoplastic lesions, of which 50% were benign, 23.08% were malignant and 26.92% presented both types of neoplasms. The remaining 43.48% of the bladders did not show any neoplasms. Presence of BPV-2 was confirmed in 43.48% of the bladders: 65% of these were found in neoplastic lesions and 35% in non-neoplastic lesions. Among the neoplastic lesions, detection of BPV-2 was more frequent in benign lesions (53.84%), followed by malignant lesions (23.08%) and by those presenting both benign and malignant lesions (23.08%). These results confirm the presence of BPV-2 in both neoplastic and non-neoplastic conditions in urinary bladders of cattle with enzootic hematuria from the studied geographic area.

PCR Amplification of DNA Sequence of Bovine Papillomavirus Type 2 in Urinary Bladder of Cattle with Enzootic Hematuria in Espirito Santo, Brazil

The goal of this study was to investigate the presence of bovine papillomavirus type 2 (BPV-2) genomic DNA using polymerase chain reaction (PCR) assays, in the urinary bladder of cattle with bovine enzootic hematuria (BEH), in the neoplastic and non-neoplastic lesions in Espírito Santo, Brazil. Thus, 46 bladders with gross BEH lesions were collected from adult cattle at slaughterhouses. The samples were divided into two portions: one for use in histopathological analysis and the other one for PCR. Microscopic analysis showed that 56.52% of the bladders presented neoplastic lesions, of which 50% were benign, 23.08% were malignant and 26.92% presented both types of neoplasms. The remaining 43.48% of the bladders did not show any neoplasms. Presence of BPV-2 was confirmed in 43.48% of the bladders: 65% of these were found in neoplastic lesions and 35% in non-neoplastic lesions. Among the neoplastic lesions, detection of BPV-2 was more frequent in benign lesions (53.84%), followed by malignant lesions (23.08%) and by those presenting both benign and malignant lesions (23.08%). These results confirm the presence of BPV-2 in both neoplastic and non-neoplastic conditions in urinary bladders of cattle with enzootic hematuria from the studied geographic area.(AU)

Chenopodium ambrosioides L. essential oil and ethanol extract on control of canine Ancylostoma spp / Extrato etanólico e óleo essencial de Chenopodium ambrosioides L. no controle de Ancylostoma spp. de cães

The use of Chenopodium ambrosioides L. has shown to be promising in the management of gastrointestinal nematodes. The objective of this study was to quantitate the yield and characterize the chemical composition of the essential oil of C. ambrosioides, as well as to evaluate the in vitro effect of the ethanolic extract and the essential oil in L3 of Ancylostoma spp. and the in vivo effect(s) of the essential oil in dogs. The effects of the ethanol extract and essential oil on Ancylostoma spp. were evaluated in vitro by exposing larvae to the extract at concentrations ranging from 0.005 g mL-1 to 0.2 g mL-1 and to essential oil at concentrations of 50, 100, and 150 ?L mL-1. For the in vivo test, 26 healthy dogs, naturally infected by Ancylostoma spp., were divided into three groups: F1 - cookies were administered without active principle; F2 - herbal cookies containing 37.5 ?L g-1 essential oil of C. ambrosioides L.; F3 - cookies plus a commercial formulation containing febantel, pyrantel, praziquantel, and ivermectin. Complete blood counts and serum biochemistry for AST, ALT, AF, urea, creatinine, total protein, and albumin were performed. The yield of the essential oil was 0.3% m v-1, and its major components included ?-terpinene (1.24%), p-cymene (4.83%), and ascaridol Z (87%) and E (5.04%) isomers. The concentrations of C. ambrosioides L. ethanol extract used were ineffectiveagainst Ancylostoma spp. larvae. The essential oil at a concentration of 150 μL mL-1 was effectiveagainst L3 larvae. In the in vivo study in dogs, the herbal cookies containing C. ambrosioides L essentialoil reduced the number of eggs per gram of feces.

O uso de Chenopodium ambrosioides L. tem se mostrado promissor no controle de nematoides gastrintestinais. Objetivou-se verificar o rendimento e a composição química do óleo essencial de C. ambrosioides, assim como avaliar o efeito in vitro do extrato etanólico e do óleo essencial em L3 de Ancylostoma spp., e o efeito in vivo do óleo essencial para cães. Para a atividade larvicida as larvas de Ancylostoma spp. foram colocadas na presença do extrato etanólico nas concentrações de 0,005 g mL-1, 0,01 g mL-1, 0,02 g mL-1, 0,03 g mL-1, 0,04 g mL-1; 0,05 g mL-1, 0,06 g mL-1, 0,08 g mL-1, 0,1 g mL-1, 0,12 g mL-1 e 0,2 g mL-1, do óleo essencial nas concentrações de 50 µL mL-1, 100 µL mL-1 e 150 µL mL-1. Para o teste in vivo utilizaram-se 26 cães, hígidos, infectados naturalmente por Ancylostoma spp., divididos em três grupos: F1 - foram administrados biscoitos sem princípio ativo; F2 - biscoitos fitoterápicos com óleo essencial de C. ambrosioides L na concentração de 37,5 µL g-1; F3 biscoitos acrescido de formulação comercial a base de febantel, pirantel, praziquantel e ivermectina. Foi realizado hemograma completo e bioquímica sérica para AST, ALT, FA, ureia, creatinina, proteína total e albumina. Verificou-se o rendimento de 0,3% m v-1 do óleo essencial e na sua composição química a presença de ?-terpineno (1,24 %), o p-cimeno (4,83 %) e ascaridol, nos seus isômeros Z (87%) e E (5,04 %), como componentemajoritário. O extrato etanólico de C. ambrosioides L., nas concentrações utilizadas, apresentouinsuficiente atividade para o efeito larvicida. O óleo essencial na concentração de 150 µL mL-1 foi eficazcontra L3. A formulação de biscoito fitoterápico com óleo essencial C. ambrosioides L reduziu o númerode ovos por grama de fezes dos cães.

Chenopodium ambrosioides L. essential oil and ethanol extract on control of canine Ancylostoma spp. / Extrato etanólico e óleo essencial de Chenopodium ambrosioides L. no controle de Ancylostoma spp. de cães

The use of Chenopodium ambrosioides L. has shown to be promising in the management of gastrointestinal nematodes. The objective of this study was to quantitate the yield and characterize the chemical composition of the essential oil of C. ambrosioides, as well as to evaluate the in vitro effect of the ethanolic extract and the essential oil in L3 of Ancylostoma spp. and the in vivo effect(s) of the essential oil in dogs. The effects of the ethanol extract and essential oil on Ancylostoma spp. were evaluated in vitro by exposing larvae to the extract at concentrations ranging from 0.005 g mL-1 to 0.2 g mL-1 and to essential oil at concentrations of 50, 100, and 150 ?L mL-1. For the in vivo test, 26 healthy dogs, naturally infected by Ancylostoma spp., were divided into three groups: F1 - cookies were administered without active principle; F2 - herbal cookies containing 37.5 ?L g-1 essential oil of C. ambrosioides L.; F3 - cookies plus a commercial formulation containing febantel, pyrantel, praziquantel, and ivermectin. Complete blood counts and serum biochemistry for AST, ALT, AF, urea, creatinine, total protein, and albumin were performed. The yield of the essential oil was 0.3% m v-1, and its major components included ?-terpinene (1.24%), p-cymene (4.83%), and ascaridol Z (87%) and E (5.04%) isomers. The concentrations of C. ambrosioides L. ethanol extract used were ineffectiv

O uso de Chenopodium ambrosioides L. tem se mostrado promissor no controle de nematoides gastrintestinais. Objetivou-se verificar o rendimento e a composição química do óleo essencial de C. ambrosioides, assim como avaliar o efeito in vitro do extrato etanólico e do óleo essencial em L3 de Ancylostoma spp., e o efeito in vivo do óleo essencial para cães. Para a atividade larvicida as larvas de Ancylostoma spp. foram colocadas na presença do extrato etanólico nas concentrações de 0,005 g mL-1, 0,01 g mL-1, 0,02 g mL-1, 0,03 g mL-1, 0,04 g mL-1; 0,05 g mL-1, 0,06 g mL-1, 0,08 g mL-1, 0,1 g mL-1, 0,12 g mL-1 e 0,2 g mL-1, do óleo essencial nas concentrações de 50 µL mL-1, 100 µL mL-1 e 150 µL mL-1. Para o teste in vivo utilizaram-se 26 cães, hígidos, infectados naturalmente por Ancylostoma spp., divididos em três grupos: F1 - foram administrados biscoitos sem princípio ativo; F2 - biscoitos fitoterápicos com óleo essencial de C. ambrosioides L na concentração de 37,5 µL g-1; F3 biscoitos acrescido de formulação comercial a base de febantel, pirantel, praziquantel e ivermectina. Foi realizado hemograma completo e bioquímica sérica para AST, ALT, FA, ureia, creatinina, proteína total e albumina. Verificou-se o rendimento de 0,3% m v-1 do óleo essencial e na sua composição química a presença de ?-terpineno (1,24 %), o p-cimeno (4,83 %) e ascaridol, nos seus isômeros Z (87%) e E (5,04