Submitted by Ana Maria Fiscina Sampaio (fiscina@bahia.fiocruz.br) on 2014-11-17T19:30:12Z No. of bitstreams: 1 Alene Vanessa Azevedo dos Santos. Reacoes de oxido reducao como alvo de quimioterapia triconomicida.pdf: 13103565 bytes, checksum: ad5cc355a7ef3d05b87bd59e85b6f9e0 (MD5) / Made available in DSpace on 2014-11-17T19:30:12Z (GMT). No. of bitstreams: 1 Alene Vanessa Azevedo dos Santos. Reacoes de oxido reducao como alvo de quimioterapia triconomicida.pdf: 13103565 bytes, checksum: ad5cc355a7ef3d05b87bd59e85b6f9e0 (MD5) Previous issue date: 2008 / Fundação Oswaldo Cruz. Centro de Pesquisa Gonçalo Moniz. Salvador, BA, Brasil / Trichomonas vaginalis e Tritrichomonas foetus são os agentes etiológicos da tricomoníase humana e bovina, respectivamente. A primeira é uma parasitose de grande importância em saúde pública, sendo a principal doença sexualmente transmissível não viral e a segunda ocasiona extensos prejuízos econômicos na pecuária. A tricomoníase humana pode promover a incidência de câncer de colo de útero e a transmissão do HIV. Na infestação humana a droga de escolha é o metronidazol, mas não há um composto eficiente para o tratamento de bovinos. Como este fármaco é usado para outras parasitoses e infecções bacterianas desde a década de 60, tem sido documentado o surgimento de casos refratários e cepas resistentes. Além disso, o fármaco pode ter efeitos mutagênicos e carcinogênicos. Assim sendo a busca por novos compostos tricomonicidas constitui uma demanda premente. No presente estudo foram avaliados os efeitos do dietilditiocarbamato de sódio (DETC) sobre os protozoários parasitas T. vaginalis e T. foetus. Observamos que a adição de DETC inibiu significativamente a proliferação de T. vaginalis, produzindo uma IC50 de 269,7nM, enquanto o metronidazol teve uma IC50 de 523,1nM. Em T. foetus os valores de IC50 foram semelhantes, sendo 497,8 e 459,7nM para o DETC e para o metronidazol, respectivamente. Estes compostos não afetaram significativamente a incorporação de [3H]timidina por esplenócitos murinos em concentrações de até 1000M. A atividade mitocondrial de macrófagos peritoneais murinos também não foi afetada por DETC, mesmo em 200μM, sugerindo seletividade no modo de ação. Objetivando determinar a existência de sinergismo entre DETC e metronidazol, foram determinados os valores de FIC (concentração inibitória fracionada) para os dois protozoários. Os valores de FIC foram 0,3 e 0,7 para T. vaginalis e T. foetus, respectivamente. Esta observação indica que ocorre a interação sinergística no protozoário parasita de humanos. A fim de determinar se a incubação com DETC poderia afetar a expressão de grupos sulfidrila dos parasitos empregamos a reação de Ellman para dosar os grupamentos tiol totais de T. vaginalis antes e após a adição do composto. Observamos que o tratamento com DETC por 24h significativamente (p<0,01) reduziu a concentração de grupos tióis do parasito. A detecção de tióis livres pela sonda fluorescente orto-phthalaldeído (OPA) em T. foetus sugere a participação de sulfidrilas no mecanismo de ação do DETC, uma vez que este reduz marcadamente a marcação dos parasitos pelo OPA, mas este efeito foi revertido pela pré-incubação com cisteína. Vale salientar que o aminoácido reverte, ainda, os efeitos do composto na proliferação parasitária. A mensuração de radicais livres por quimioluminescência amplificada pelo luminol indicou que a geração de espécies reativas de oxigênio foi significativamente (p <0,05) aumentada por DETC em T. vaginalis, mas não em T. foetus. O estresse oxidativo sobre os parasitas foi avaliado pela medida de substâncias reativas ao ácido tiobarbitúrico (TBARs). Observou-se que a combinação metronidazol-DETC significativamente aumenta a peroxidação lipídica em T. vaginalis (p <0,01) e T. foetus (p <0,05). A análise ultraestrutural por microscopia eletrônica de transmissão revelou que tanto DETC quanto o metronidazol produziram danos hidrogenossomais e desencadearam autofagia e esses efeitos foram mais acentuadas no parasitas incubados com a combinação de drogas. Tomados em conjunto, estes dados sugerem que a combinação metronidazol-DETC pode fornecer novas ferramentas para a efetiva quimioterapia da tricomoníase / Trichomonas vaginalis e Tritrichomonas foetus are the etiologic agents of the human and bovine trichomoniasis, respectively. The former is a parasitic disease of great relevance in public health – the major non-viral sexually-transmitted disease, whereas the latter causes remarkable losses in livestock productivity. Human trichomoniasis can promote the incidence of cervical cancer and HIV transmission. In human infestation the drug of choice is metronidazole, but there is no efficient compound for treating the cattle. Since the medication is also used for other parasitic and bacterial diseases, since the 60s, refractory cases and resistant strains have been documented. Besides, the drug may be mutagenic and carcinogenic. Therefore the search for new trichomonicidal compounds is required. In the present study we investigated the effects of sodium N,N-diethylthiolcarbamate (DETC) alone or combined with metronidazole upon the parasitic protozoa T. vaginalis and T. foetus. We notice that DETC significantly inhibited the T. vaginalis proliferation, producing an IC50 of 269.7nM, whereas metronidazole produced an IC50 of 523.1. In T. foetus the IC50 values were similar, being 497.8 and 459.7nM for DETC and metronidazole, respectively. These compounds did not significantly affect the incorporação de [3H]timidine incorporation by murine splenocytes in concentrations up to 1000μM. The mitochondrial activity of murine peritoneal macrophages was not affected by 200μM DETC, suggesting a selective mode of action. In order to determine whether there is synergism between DETC and metronidazole, we determined the fractional inhibitory concentrations (FIC) of these compounds upon both protozoa. The FIC values for T.vaginalis and T. foetus were 0.3 and 0.7, respectively. This observation indicates that synergistic interaction takes place only on the human pathogen. To determine whether the incubation with DETC could affect the expression of sulphydril groups of parasites we employed the Ellman’s reaction to measure the total thiol groups of T. vaginalis before and after the addition of the compound. We observed that treatment with DETC for 24h significantly (p <0.01) reduced the concentration of thiol groups of the parasite. The detection of free thiols by the fluorescent probe ortho- phthaldialdehyde (OPA) in T. foetus suggests the involvement of sulphydrils in the DETC mechanism of action, since it markedly reduces the OPA labeling of parasites, but this effect was reversed by cysteine preincubation with. It is noteworthy that the amino acid addition also reverts, the effects of the compound on the parasite proliferation. The measurement of free radicals by luminol-enhanced chemiluminescence indicated that reactive oxygen species generation was significantly (p <0.05) enhanced by DETC in T. vaginalis, but not T. foetus. The oxidative stress on the parasites was evaluated by measurement of the thiobarbituric acid-reactive substances (TBARs). We observed that the metronidazole-DETC combination significantly enhanced lipid peroxidation in T. vaginalis (p <0.01) and T. foetus (p <0.05). The ultrastructural analysis by transmission electron microscopy revealed that both DETC and metronidazole produced hydrogenosomal damage and triggered autophagy and these effects were more pronounced on the parasites incubated with the combined drugs. Taken together these data suggest that the metronidazole-DETC combination may furnish new tools in the effective chemotherapy of trichomoniasis.

Infectious disease continues to cause significant problems on reproductive efficiency in the cattle industry. The purpose of this project is to evaluate new testing strategies for Tritrichomonas foetus and Campylobacter fetus subsp. venerealis. This thesis describes the result of three studies that evaluated the use of real-time PCR for the identification of Tritrichomonas foetus and Campylobacter fetus subsp. venerealis in carrier bulls. The first study evaluated the specificity of a real-time PCR test for T. foetus in individual culture enriched samples, and the sensitivity of the assay for use in pooled samples of up to 25 bulls. Specificity estimates were 98.8% (95% CI 97-99.4) and 100% (95% CI 98.9-100) for culture and real-time PCR, respectively. The sensitivity of the real-time PCR assay for pooled preputial samples was: 96.8% (83.8-99.4) for pool ratios 1/3 and 1/5; 93.5% (79.3-98.2) for pool ratios 1/2, 1/15, 1/20 and 1/25; and 90.3% (75.1-96.6), and were not significantly different. However, 13 of the 217 pools tested were negative and 9 of these negative testing pools contained the same positive sample. The media in this positive sample showed evidence of contamination and could potentially explain the failure to detect T. foetus. The second study evaluated the sensitivity of a real-time PCR for the detection of T. foetus in individual and pooled direct preputial samples. Sensitivity of individual samples tested by culture, real-time PCR in direct and culture enriched samples were determined from 121 samples obtained from 9 infected bulls. Sensitivity estimates were: 95.0% (95% CI: 89.6% to 97.7%) for culture, 95.9% (95% CI: 90.7 to 98.2) for real-time PCR in cultured enriched samples, and 90.1% (95% CI: 83.5 to 94.2) for direct preputial samples and did not differ (P=0.12). Sensitivity estimates for direct pooled samples in groups of 5 or 10 were: 83.6% (95% CI: 75.6 to 89.4) and 77.3% (95% CI: 68.6-84.1), respectively and were not significantly different (P=0.08). The use of repeat sampling tested in pools by real-time PCR increased the sensitivity to 100% and 96% for 3 consecutive samples (pools of 5 or 10, respectively). The use of pooled direct preputial samples although sensitive, still requires the use of repeated sampling. The third study determined the sensitivity and specificity of a recently developed real-time PCR (qPCR) tests for Cfv. A total of 300 virgin bulls were tested by both culture and qPCR. Specificity estimates were 85% (95% CI: 80.5 to 88.6) for qPCR and 100% (95% CI: 98.7 to 100) for culture, and were significantly different (P<0.01). A total of 4 naturally infected bulls and 9 artificially infected bulls were sampled serially to obtain positive samples for a sensitivity analysis. Sensitivity estimates and 95% confidence intervals are as follows: qPCR (85.4%, 95% CI: 80.6-89.2); direct culture on blood agar (82.3%, 95% CI: 77.2-86.5), DFAT (72.1%, 95% CI: 66.2-77.4), direct culture on Skirrow agar (32.7%, 95% CI: 27.2-38.7), TEM and blood agar (30%, 95% CI: 23.4-37.5), and TEM and Skirrow agar (38.1%, 95% CI: 31-45.9). The sensitivity of the different tests evaluated varied significantly with different ambient temperatures (P<0.01). The sensitivity of the qPCR was significantly higher than any other test when temperatures exceeded 5°C. The use of repeated sampling at weekly intervals significantly improved the sensitivity of the qPCR. The real-time PCR assay for the detection of T. foetus in both individual and pooled samples appears to be highly sensitive and specific. Moreover, the possibility of using direct preputial samples provides a cost-effective diagnostic strategy. Real-time PCR in direct preputial samples for BGC diagnosis in bulls has good sensitivity and specificity. However, the use of repeated sampling maybe needed in order to maximize the ability to detect carrier bulls.

Trichomonosis is currently the most important venereal disease of cattle in South Africa with adverse economic implications to the beef production industry due to cow abortions, infertility and culling of carrier bulls. Once diagnosed in a herd, eradication is difficult due to financial and biological implications. Bulls are asymptomatic carriers and susceptibility increases with age. In infected females, clinical signs include embryonal death, abortion, pyometra, foetal maceration and uterine discharge. Diagnostic accuracy is one of the major clinical problems preventing easy eradication of trichomonosis from a herd and can be influenced by biological variance in the occurrence of the organism, sampling errors, sample degradation during sample transport and diagnostic laboratory inaccuracies. This study aimed to validate the accuracy of voluntarily enrolled private (n = 8) and state-owned (n = 5) laboratories that perform trichomonosis diagnostic tests by estimating the sensitivity (Se) and specificity (Sp) per laboratory. It was hypothesized that diagnostic laboratories in South Africa play an insignificant role in the inaccuracy of the diagnosis of trichomonosis. Laboratories performed either the culture method (n = 5), polymerase chain reaction (PCR) (n = 6) or a combination of culture and PCR (n= 2). Fresh preputial scrapings from four bulls with known negative status for trichomonosis were pooled in 200ml of phosphate buffered saline (PBS) to form the sample base for 12 subsamples of 13ml each. Duplicate subsamples were then contaminated with 2ml originating from four different laboratory cultures of Tritrichomonas foetus or 2ml of culture medium for four negative samples. Aliquots of the subsamples were transferred to an anaerobic transport medium, and the final concentration reached in these samples submitted to the laboratories, were categorised as follows: weak (<10 organisms/μl), moderate (10 – 30 organisms/μl) or strong (>30 organisms/μl). A total of 312 samples were sent by courier in two separate rounds: eight (4 duplicates) positive and four negative samples per round. Multiple logistic regression was performed on sensitivity, using sampling round, laboratory sector, diagnostic test type and sample concentration as independent variables, and removing variables in a stepwise manner based on the highest P-value. Two public laboratories only reported on one round of sampling, and one batch of 12 samples was severely delayed in reaching another public laboratory. The sample identifications of a further two batches were not recorded by the respective private laboratories. The results from these 60 unreported samples were not included in the analysis. Laboratories that performed the PCR assay (solely, or in addition to culture) were grouped for data analysis. The overall specificity (Sp) was 100% and the sensitivity was 88.7% (95% CI 83.9% - 93.5%). Laboratories using PCR recorded higher sensitivity than those using the culture method (95.5%; 95% CI 91.0% – 99.9% and 81.3%; 95% CI 72.5% - 90.0% respectively, P < 0.01), and private laboratories recorded higher Se than public laboratories (96.4%; 95% CI 92.9% - 99.9% and 73.2%; 95% CI 61.2% - 85.2%, P < 0.01). For laboratories using PCR, weak positive samples recorded a lower sensitivity than strong positive samples (86.4%; 95% CI 70.8% - 101.9% and 100%; 95% CI 100% - 100%, respectively, P < 0.01). One public and six private laboratories obtained 100% accuracy during the two sampling rounds. In the logistic regression model, private sector (compared to public), an increasing concentration of organisms in the sample and the second round of sampling (compared to the first round) were independent predictors of laboratory sensitivity for the detection of Tritrichomonas foetus. It is concluded that inaccuracies in the diagnostic laboratory contributes to the deficiencies in diagnostic sensitivity for trichomonosis in South Africa, but does not influence diagnostic specificity. It is further concluded that diagnostic sensitivity was independently influenced by the sector in which the laboratory operates (private vs public) and the concentration of Tritrichomonas foetus organisms in the sample. / Dissertation (MSc)--University of Pretoria, 2019. / Production Animal Studies / MSc (Veterinary Science) / Unrestricted

La tritrichomonosis bovina (TB) es una enfermedad de transmisión sexual causada por el protozoo Tritrichomonas foetus que produce muerte embrionaria, piómetra y abortos esporádicos. El objetivo de esta tesis fue estudiar los mecanismos involucrados en la pérdida del conceptus en un modelo murino de TB. Se infectaron ratonas BALB/c con T. foetus, que fueron apareadas con machos sanos. Se utilizaron técnicas histológicas para estudiar los cambios endometriales en las ratonas infectadas y analizar el número y distribución de las células natural killer uterinas y de los mastocitos. Mediante lectinahistoquímica e inmunohistoquímica se analizaron los carbohidratos y la muerte y proliferación del epitelio uterino, respectivamente. Con real time RT-PCR se estudió la expresión de citoquinas, hemooxigenasa-1 (HO-1) e indolamina 2,3 dioxigenasa (IDO) y a las células T reguladoras (Treg) y T helper 17 (Th17), en el útero. En el grupo de ratonas infectadas, la muerte embrionaria se produjo en las fases temprana y media de la gestación, asociada a endometritis y cambios en el patrón de carbohidratos y en los índices de proliferación y muerte celular del epitelio uterino. El ARNm de TNF-α, IL-10 e IL-4 aumentó en el grupo infectado. La expresión de Foxp3, un gen relacionado con las células Treg, y RORγt, marcador de células Th17, fue mayor en el útero de las ratonas infectadas. Los niveles de ARNm de HO-1 disminuyeron en las ratonas que sufrieron pérdida embrionaria. Los cambios en el patrón de carbohidratos y en los índices de muerte y proliferación celular, así como las respuestas T de tipo 1 y 17 podrían estar involucradas en la muerte embrionaria producida durante la infección con T. foetus. Estas respuestas alterarían los mecanismos protectores de la preñez como HO-1. El aumento de las células Treg promovería la persistencia de la infección por su acción inmunosupresora. De esta manera, en el modelo de ratonas preñadas, se caracterizaron mecanismos inmunológicos relacionados con la muerte embrionaria temprana en la TB.

The aim of this work was to examine the effects of sampling method on accuracy of culture for Tritrichomonas foetus; and the effects of sampling method, time delay, and addition of a DNA preservative on the accuracy of a PCR test. Samples from two different sources were used for Experiment 1. Preputial scrapings were collected from one group of three infected and one uninfected bull 10 times. Secondly, samples were collected from 5 infected bulls by both sheath washing and scraping on 6 occasions, while 8 uninfected animals were sampled three or more times. Twenty nine out of 30 samples from the first sample set were found to be positive, and 83 % of samples collected by both methods for the second sample set tested positive. No samples from the control bulls were found to be positive. Scraping was found to offer significant practical advantages over washing. It may be subject to greater operator variability than sheath washing. The second experiment utilised the same samples as were used for the second data set under Experiment 1. Guanidinium thiocyanate (GuSCN) was added to half of each sample. Each sample was cultured, while all samples were subjected to DNA extraction within 6 and 30 hours and after 5 days of storage at 4 °C. PCR and agarose gel electrophoresis was performed. No samples from the control animals tested positive on PCR. The sensitivity of the PCR on samples from infected bulls ranged from 0,9 in samples extracted within 6 hours to 0,31 in samples extracted after 5 days. Sampling method had no effect with the exception of samples held for 5 days with GuSCN, where sheath washing was superior to scraping. The addition of GuSCN had no effect. Holding time reduced sensitivity at 5 days, but the effect was not significant at 30 hours. It is concluded that preputial scraping is equal in sensitivity to washing for culture of Tritrichomonas foetus. Preputial samples for PCR testing should be submitted as soon as possible after collection, and the addition of GuSCN has no effect. Samples collected by sheath washing may be superior to those collected by scraping for PCR testing. The requirement for a test with sufficient sensitivity to allow reliable identification of infected bulls based on one sample has not been met with the described method. / Dissertation (MMedVet (Gyn))--University of Pretoria, 2002. / Production Animal Studies / unrestricted

Resumo:O presente estudo investigou a ocorrência de Tritrichomonas foetus em gatos no Brasil. T. foetus é um protozoário que em felinos coloniza o cólon causando colite. O sinal clínico é a diarreia crônica do intestino grosso. Gatos infectados podem apresentar resolução espontânea da diarreia, mas podem permanecer infectados pela vida toda. Existem três métodos diagnósticos mais difundidos que identificam a presença de T. foetus nas fezes do animal, sendo: o exame direto das fezes utilizando o microscópio óptico, a cultura e a PCR. No presente estudo, foram comparados dois métodos diagnósticos, o exame direto das fezes e a PCR. Também foi realizado o sequenciamento genético. Foram colhidas amostras fecais de 129 gatos por meio da técnica de lavado retal. Cada amostra foi examinada ao microscópio óptico. A presença de DNA do T. foetus foi verificada pela PCR através da amplificação de 347 pares de bases a partir dos primers TFR3 e TFR4. Amplicons dos casos positivos foram sequenciados. T. foetus foi observado em uma amostra através do exame diretos das fezes, enquanto que a PCR foi positiva para cinco gatos (6,45%). À análise estatística não houve correlação significativa entre positividade para T. foetus e sexo, idade, raça, presença de diarreia, histórico de diarreia, tratamento prévio, estilo de vida, ambiente, origem dos animais e presença de coinfecções. O isolado de T. foetus mostrou 100% de similaridade com outros isolados do parasito de felino no mundo e revelou polimorfismo num único nucleotídeo (T>C) quando comparado com o isolado de bovino. Devido ao crescente interesse sobre este parasito em gatos, o presente estudo visa contribuir com futuros registros da ocorrência do T. foetus em gatos visto que se trata do primeiro estudo sobre sua ocorrência no Brasil / Abstract:The present study investigated the occurrence of T. foetus in cats from Brazil. T. foetus is a protozoan that in cats it colonizes the colon resulting in colitis. The major clinical sign is chronic large bowel diarrhea. Infected cats may have spontaneous resolution of diarrhea, but they can remain infected for all life. In veterinary medical practice, there are three most widespread diagnostic methods that identify T. foetus in animal feces: direct examination of feces under the microscope, culture and PCR. In the present study, it was compared two diagnosis methods, direct examination of feces and PCR. And also it was done the genetic sequencing. Fecal samples from 129 cats were collected by rectal flush technique. Each sample was examined by optical microscopy (direct examination). The presence of T. foetus DNA was verified using PCR by amplification of 347 bases pairs from the primers TFR3 and TFR4. Amplicons of positive cases were sequenced. T. foetus was observed in one sample by direct microscopic examination of feces while PCR was positive in five cats (6.45%). Statistical analyses showed no significant associations between T. foetus infection and sex, age, breed, presence of diarrhea, history of diarrhea, previous treatment, lifestyle, environment, origin of the animals, and co-infections. The isolate of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world and revealed a single nucleotide polymorphism (T>C) when compared with T. foetus isolate from cattle. Due the increasing interest in this parasite in cats, the present study contributes in further reporting the worldwide occurrence of T. foetus in cats because it is the first study about its occurrence in Brazil. / Orientador:Marcelo Vasconcelos Meireles / Banca:Rosangela Zacarias Machado / Banca:Gosele Fabrino Machado / Mestre

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-05-03T19:48:33Z No. of bitstreams: 1 2016 - Lara Patricia Santos Carrasco.pdf: 1457762 bytes, checksum: 82c38851cf4f7b3e95d40cfbb6f7ddee (MD5) / Made available in DSpace on 2018-05-03T19:48:33Z (GMT). No. of bitstreams: 1 2016 - Lara Patricia Santos Carrasco.pdf: 1457762 bytes, checksum: 82c38851cf4f7b3e95d40cfbb6f7ddee (MD5) Previous issue date: 2016-02-16 / Tritrichomonas foetus has been recently recognized as an important cause of diarrhea in various countries. The aim of this study was to diagnose and treat the infection by T. foetus in cats treated at HVPA of UFRRJ, identify the clinical relevance of this parasite, the risk factors, presence of co-infections and clinical response to treatment. In this study, were admitted 100 felines, from different breeds, both sexes and from 16 weeks of age. As inclusion criteria, patients had to have diarrhea history for more than three weeks, with little success of clinical response to treatments previously used. From the stool samples obtained by colon washing or spontaneous defecation it was conducted direct examination of stool, centrifugal flotation technique and real-time PCR. Of the 100 cats selected for this study, 10% had T. foetus infection. Most positive animals were over one year old (70%) and only 30% had up to 1 year old. Cats were 80% purebred and 20% were animals mongrel. The clinical signs present were diarrhea (60%), weight loss (10%), hemoatochezia (30%) and presence of mucus in the stool (20%). Only two animals were positive for the antigen of FeLV virus, and theses showed more severe case of diarrhea. Three animals showed co-infections with other enteropathogens (30%), one with Giardia, one with Cryptosporidium and one with Cystoisospora. All treated animals showed clinical improvement with the administrations of ronidazole at a dose 30mg/kg every 24 hours during 14 days and showed no recurrence until the recent time. We can conclude that T. foetus infection is an important differential diagnosis in cats with chronic diarrhea, which cats of any age and regardless of whether purebred or not, may have the infection, other enteropathogens needs to be investigated in all cases and the treatment was effective in the resolution of clinical signs of all treated cats. / Tritrichomonas foetus foi recentemente reconhecido como um importante agente etiol?gico de diarreia em diversos pa?ses. O objetivo deste estudo foi diagnosticar e tratar a infec??o por T. foetus nos gatos atendidos no HVPA da UFRRJ, identificar a relev?ncia cl?nica deste parasita, os fatores de risco, a presen?a de co- infec??es e resposta cl?nica ao tratamento. Neste trabalho, foram admitidos 100 animais da esp?cie felina, de diferentes ra?as, de ambos os sexos e a partir de 16 semanas de idade. Como crit?rio de inclus?o, os pacientes deveriam apresentar hist?rico de diarreia a mais de tr?s semanas, com pouco sucesso de resposta cl?nica para terap?uticas previamente institu?das. A partir das amostras de fezes obtidas por coleta via lavagem do colon dos animais e pelo ato de defeca??o espont?nea, foram realizados os m?todos do exame direto das fezes, t?cnica de centrifugo flutua??o e PCR em tempo real. Dos 100 gatos que foram selecionados para esta pesquisa, 10% apresentaram infec??o por T. foetus. A maioria dos animais positivos estavam na faixa et?ria acima de um ano (70%) e apenas 30% tinha at? um ano de idade. Os gatos eram 80% de ra?a pura e 20% eram animais sem ra?a definida. Os sinais cl?nicos apresentados por esses gatos foram diarreia (60%), perda de peso (10%), hematoquezia (30%) e presen?a de muco nas fezes (20%). Apenas dois animais infectados eram positivos para o ant?geno do v?rus da FeLV, e estes apresentaram um quadro mais severo de diarreia. Tr?s animais apresentaram coinfec??es com outros enteropat?genos (30%), um com Giardia sp, um com Cryptosporidium e outro com Cystoisospora. Todos os gatos tratados apresentaram melhora cl?nica com a administra??o do f?rmaco ronidazol na dose de 30 mg/Kg a cada 24 horas por 14 dias, n?o apresentando recidiva da diarreia at? o recente momento. Podemos concluir que a infec??o por T. foetus ? um importante diagn?stico diferencial em felinos com diarreia cr?nica, que gatos de qualquer idade e independente de ser de ra?a pura ou n?o, podem apresentar a infec??o, que a co- infec??o com outros enteropat?genos necessita ser investigada em todos os casos e que o tratamento foi efetivo na resolu??o dos sinais cl?nicos de todos os gatos tratados.

Made available in DSpace on 2016-09-27T13:39:59Z (GMT). No. of bitstreams: 0 Previous issue date: 2015-12-01. Added 1 bitstream(s) on 2016-09-27T13:45:08Z : No. of bitstreams: 1 000870641.pdf: 755047 bytes, checksum: 904fea33f34db16ae9b0d792a4458f19 (MD5) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / The present study investigated the occurrence of T. foetus in cats from Brazil. T. foetus is a protozoan that in cats it colonizes the colon resulting in colitis. The major clinical sign is chronic large bowel diarrhea. Infected cats may have spontaneous resolution of diarrhea, but they can remain infected for all life. In veterinary medical practice, there are three most widespread diagnostic methods that identify T. foetus in animal feces: direct examination of feces under the microscope, culture and PCR. In the present study, it was compared two diagnosis methods, direct examination of feces and PCR. And also it was done the genetic sequencing. Fecal samples from 129 cats were collected by rectal flush technique. Each sample was examined by optical microscopy (direct examination). The presence of T. foetus DNA was verified using PCR by amplification of 347 bases pairs from the primers TFR3 and TFR4. Amplicons of positive cases were sequenced. T. foetus was observed in one sample by direct microscopic examination of feces while PCR was positive in five cats (6.45%). Statistical analyses showed no significant associations between T. foetus infection and sex, age, breed, presence of diarrhea, history of diarrhea, previous treatment, lifestyle, environment, origin of the animals, and co-infections. The isolate of T. foetus showed 100% identical sequences with other T. foetus isolates from cats around the world and revealed a single nucleotide polymorphism (T>C) when compared with T. foetus isolate from cattle. Due the increasing interest in this parasite in cats, the present study contributes in further reporting the worldwide occurrence of T. foetus in cats because it is the first study about its occurrence in Brazil. / FAPESP: 2011/112436

Submitted by Mario BC (mario@bc.ufrpe.br) on 2017-05-05T15:00:33Z No. of bitstreams: 1 Junior Mario Baltazar de Oliveira.pdf: 1499654 bytes, checksum: 6880a06cd3c2f0f28c7d609803cb7e9e (MD5) / Made available in DSpace on 2017-05-05T15:00:33Z (GMT). No. of bitstreams: 1 Junior Mario Baltazar de Oliveira.pdf: 1499654 bytes, checksum: 6880a06cd3c2f0f28c7d609803cb7e9e (MD5) Previous issue date: 2014-08-08 / The objective of this study was to perform an epidemiological analysis of Campylobacter fetus subsp. venerealis and Tritrichomonas foetus infection in cattle from state of Pernambuco, Brazil. For this, 383 samples of cervical-vaginal mucus of cows from 21 properties distributed in the 19 counties that make up the microregion of Garanhuns and 105 samples of preputial smegma of males in the reproductive age, coming from 32 properties and four slaughterhouses were collected from state Pernambuco. The samples had their genomic materials extracted and subjected to polymerase chain reaction for diagnosis. To analyze the risk factors applied an investigative questionnaire on the hygienic, sanitary and reproductive management. The risk factors identified in this study was to bovine genital campylobacteriosis in female herds above 100 animals (OR=7.2, C.I. 1.3 to 38.4%, p=0.020) and bovine trichomoniasis was the use of natural mating (OR=2.4, C.I. 1.1 to 5.9%, p=0.041). In males, there was a frequency of 6.6% (2.7%-13.2%, 95% C.I.) for T. fetus and no sample was positive for C. fetus subsp. venerealis. Regarding the number of outbreaks, 21.8% of farms had animals positive for T. foetus. None analyzed variables was associated with infection. Regarding the samples from slaughterhouses, none was positive. It is concluded that infection by C. fetus subsp. venerealis and T. foetus are present in cattle in the state of Pernambuco and suggest that control measures such as diagnosis and prophylaxis, and sexual segregation and rest of infected females, using a program of artificial insemination with strict health care, elimination of infected bulls and replacement for young animals are implemented to prevent the spread of agents in herds. / Objetivou-se com este trabalho realizar uma análise epidemiológica da infecção por Campylobacter. fetus subsp. venerealis e Tritrichomonas foetus em bovinos do estado de Pernambuco, Brasil. Para isso, foram colhidas 383 amostras de muco cérvico-vaginal de vacas procedentes de 21 propriedades distribuídas nos 19 municípios que compõem a microrregião de Garanhuns e 105 amostras de esmegma prepucial de machos em idade reprodutiva, procedentes de 32 propriedades e quatro matadouros do estado de Pernambuco. As amostras tiveram seu material genômico extraído e submetido à reação em cadeia da polimerase para o diagnóstico. Para análise de fatores de risco aplicou-se um questionário investigativo com perguntas referentes ao manejo higiênico-sanitário e reprodutivo. Nas fêmeas, observou-se uma prevalência de 1,8% (0,8% - 3,9%; I.C. 95%) e 33,4% (28,7% - 38,4%; I.C. 95%;) para Campylobacter fetus subsp. venerealis e Tritrichomonas foetus, respectivamente. Em relação ao número de focos observou-se que 28,6% das propriedades apresentaram pelo menos um animal positivo para C. fetus subsp. venerealis e 90,5% para T. foetus. Os fatores de risco identificados neste estudo para campilobacteriose genital bovina em fêmeas foi rebanhos acima de 100 animais (OR=7,2; I.C. 1,3 - 38,4%; p=0,020) e para tricomonose bovina foi a utilização da monta natural (OR=2,4; I.C. 1,1 - 5,9%; p=0,041). Nos machos, observou-se uma frequência de 6,6% (2,7% - 13,2%; I.C. 95%) para T. foetus e nenhuma amostra foi positiva para C. fetus subsp. venerealis. Em relação ao número de focos, 21,8% das propriedades apresentaram animais positivos para T. foetus. Nenhuma das variáveis analisadas apresentou associação com a infecção. Quanto às amostras dos matadouros, nenhuma foi positiva. Conclui-se que as infecções por C. fetus subsp. venerealis e T. foetus estão presentes nos rebanhos bovinos do estado de Pernambuco e sugere-se que medidas de controle e profilaxia como o diagnóstico, separação e repouso sexual das fêmeas infectadas, utilização de um programa de inseminação artificial com cuidados sanitários rigorosos, descarte dos touros infectados e reposição por animais jovens devem ser implementadas a fim de evitar a disseminação do agente nos rebanhos.

Submitted by Mario BC (mario@bc.ufrpe.br) on 2018-06-15T12:48:17Z No. of bitstreams: 1 Ruy Brayner de Oliveira Filho.pdf: 2028711 bytes, checksum: c66c5e9d041a3c1b673438d6c412bde8 (MD5) / Made available in DSpace on 2018-06-15T12:48:17Z (GMT). No. of bitstreams: 1 Ruy Brayner de Oliveira Filho.pdf: 2028711 bytes, checksum: c66c5e9d041a3c1b673438d6c412bde8 (MD5) Previous issue date: 2017-10-16 / Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq / The objective of this study was to determine the epidemiological situation of infection by Tritrichomonas foetus and Campylobacter fetus subsp. venerealis in cattle at Brejo Paraibano microregion, Northeastern Brazil, identifying the possible risk factors associated with T. foetus infection. For the study of T. foetus, 349 samples of cervico-vaginal mucus and smegma were collected in beef and dairy cattle (290 females and 59 males) in 31 farms and for C. fetus subsp. venerealis, 273 samples of cervico-vaginal mucus from dairy cows from 19 farms were collected. Polymerase Chain Reaction was used to identify the DNA of the agents and culture with Modified Diamond medium for isolation of T. foetus. For analysis of risk factors associated with Tritrichomonas foetus infection in bovine species, univariate analysis of the variables of interest was performed by the Pearson chi-squared test, or Fisher's exact test, when necessary. Subsequently, logistic regression was performed considering PCR (positive or negative) as the dependent variable for trichomoniasis. Thematic maps were prepared with prevalence distributions in the studied area. The prevalence of C. fetus subsp. venerealis infection in cows was 7.7% (21/273) (CI 95% 4.8% – 11.5%), and 31.6% (6/19) of the farms showed at least one positive animal. The prevalence of T. foetus infection was 3.7% (13/349) (CI 95%, 2.1 - 6.4%). Regarding the gender, a prevalence of T. foetus infection of 4.5% (females) and 0.0% (males) was observed. The percentage of farms that had at least one positive animal for T. foetus was 19.3% (6/31). In culture, no samples were positive for T. foetus. The risk factor associated with T. foetus infection identified in this study was contact of females with bulls of other farms (OR 5.9; CI 1.5 - 22.4). This is the first report of C. fetus subsp. venerealis infection in dairy cows in this region of Brazil and the first in Brejo Paraibano microregion that analyzed T. foetus infection in cattle. To reduce risk of infection, it is recommended to adopt an artificial insemination program on the farms with negative bull semen, as well as a vaccination program against Bovine Genital Campylobacteriosis to stimulate immunity to reduce the occurrence of infection and possible reproductive problems. Contact of females with males of other farms should be avoided, especially when the health status of these bulls is not known regarding these two infections. / Objetivou-se com este estudo determinar a situação epidemiológica das infecções por Tritrichomonas foetus e Campylobacter fetus subsp. venerealis em bovinos na microrregião do Brejo Paraibano, região Nordeste do Brasil, identificando os possíveis fatores de risco associados à infecção por T. foetus. Para pesquisa de T. foetus foram coletadas 349 amostras de muco cérvico vaginal e esmegma em bovinos de corte e leite (290 fêmeas e 59 machos) em 31 propriedades e para pesquisa de C. fetus subsp. venerealis foram coletadas 273 amostras de muco cérvico vaginal de vacas leiteiras procedentes de 19 propriedades. Para a pesquisa do DNA dos agentes utilizou-se a Reação em Cadeia da Polimerase e cultivo com meio Diamond Modificado para isolamento de T. foetus. Para análise dos fatores de risco associados à infecção por Tritrichomonas foetus na espécie bovina, foi realizada uma análise univariada das variáveis de interesse pelo teste qui-quadrado de Pearson, ou Exato de Fisher, quando necessário. Posteriormente foi realizada uma regressão logística considerando como variável dependente para Tricomonose a PCR (positivo ou negativo). Foram elaborados mapas temáticos com as distribuições das prevalências na área estudada. A prevalência da infecção por C. fetus subsp. venerealis em vacas foi 7,7% (21/273) (I.C. 95%; 4,8 - 11,5%) e 31,6% (6/19) das propriedades apresentaram ao menos um animal positivo. A prevalência da infecção por T. foetus foi 3,7% (13/349) (I.C. 95%; 2,1 – 6,4%). Em relação ao sexo, observou-se uma prevalência da infecção por T. foetus de 4,5% (fêmeas) e 0,0% (machos). A porcentagem de propriedades que apresentaram ao menos um animal positivo para T. foetus foi de 19,3% (6/31). No cultivo, nenhuma amostra foi positiva para T. foetus. O fator de risco associado à infecção por T. foetus identificado neste estudo foi contato de fêmeas com touros de outras propriedades (OR 5,9; I.C. 1,5 – 22,4). Este é o primeiro relato da infecção por C. fetus subsp. venerealis em vacas leiteiras nesta região do Brasil e o primeiro na microrregião do Brejo Paraibano a analisar a infecção por T. foetus em bovinos. Para diminuir os riscos de infecção, recomenda-se a adoção de um programa de inseminação artificial nas propriedades com sêmen de touros negativos, bem como um programa de vacinação contra a Campilobacteriose Genital Bovina para estimular a imunidade, com o intuito de reduzir a ocorrência da infecção e possíveis problemas reprodutivos. O contato das fêmeas com machos de outras propriedades deve ser evitado, principalmente quando não se conhece o status sanitário desses touros em relação a estas duas infecções.