RESUMO

There are no recommended drugs to treat cattle infected with the protozoan Tritrichomonas foetus (TF). Ivermectin, widely used in the treatment of intestinal parasites, was found effective against some protozoa growing in vitro. Here, its effectiveness against a TF line was investigated. Trophozoites were incubated in media with increasing concentrations of ivermectin and mortality was determined after 24 h. Ivermectin killed cells with a mean maximum effective concentration (EC50) of 2.47 µg mL-1. The effective concentration of ivermectin was rather high for a formulation suitable for systemic treatment. However, topical treatment of animals against TF could still be considered and tested.

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Bovine genital campylobacteriosis (BGC) and bovine trichomonosis (BT) are sexually transmitted diseases (STDs) that affect bovine breeding herds, decreasing their reproductive efficiency. The objective of this work was to estimate the prevalence of these diseases and their temporal-spatial distribution in the province of Formosa, Argentina. The cross-sectional study conducted between 2018 and 2021 included a total of 15,571 bulls, inter-herd prevalence being 29.62% and 17.23% for BGC and BT, respectively. The prevalence of positive animals was 2.05% for BGC and 0.43% for BT. The temporal-spatial analysis of BGC showed two distinct spatial groupings, one group had a low risk of contracting the disease (RR = 0.13; p < 0.001; 2018-2021) while the other group had a high risk (RR = 2.84; p < 0.001; 2020-2021). BT had a high-risk group for the disease (RR = 35.24; p < 0.001; 2019). This study shows that STDs are endemic in the region, providing updated and valuable information as a tool for the health management of these diseases.

RESUMO

Taguchi method was used to optimize loop mediated isothermal amplification tests aimed to amplify segments of the elongation factor 1a1 (tf-ef1a1), the 5,8 ribosomal gene (tf-5,8 r) or the beta tubulin 2 (tf-btub2) from the protozoan parasite Tritrichomonas foetus. L9 orthogonal array and quadratic loss functions that penalize deviations from prediction values revealed the effect of amplification reaction components. Analysis of variance (ANOVA) decomposed the contribution of individual factors to a small Ct. Confirmation experiments established that optimum conditions were predictable, verifiable and reproducible. Primers concentration conditioned the non-specific amplification of tf-ef1a1 while betaine and magnesium concentration contributed to accelerate the time to reach a positive threshold in tf-ef1a1, tf-5,8 r and tf-btub2. The general strategy of simple and robust experimental design holds potential as a general optimization protocol for LAMP tests in every diagnostic laboratory.

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This work describes a specimen of Chimango Caracara (Milvago chimango - Vieillot, 1816), from southern Brazil, as a new natural host for Trichomonas gallinae (Rivolta, 1878). Caseous oral lesions were observed in a young bird, and the parasite was isolated in modified Diamond's media. Morphology of the parasite was evaluated through microscopy and subsequently, sequencing of the internal transcribed spacer 1 (ITS1) of ribosomal DNA was performed to confirm T. gallinae identification. As far as authors are concerned, this is the first report of Milvago chimango as a natural host for T. gallinae.

RESUMO

Abstract Tritrichomonas foetus is a parasite that has been definitively identified as an agent of trichomonosis, a disease characterized by chronic diarrhea. T. foetus colonizes portions of the feline large intestine, and manifests as chronic and recurrent diarrhea with mucus and fresh blood, which is often unresponsive to common drugs. Diagnosis of a trichomonad infection is made by either the demonstration of the trophozoite on a direct fecal smear, fecal culture and subsequent microscopic examination of the parasite, or extraction of DNA in feces and amplification by the use of molecular tools. T. foetus is commonly misidentified as other flagellate protozoa such as Giardia duodenalis and Pentatrichomonas hominis. Without proper treatment, the diarrhea may resolve spontaneously in months to years, but cats can remain carriers of the parasite. This paper intends to serve as a source of information for investigators and veterinarians, reviewing the most important aspects of feline trichomonosis, such as trichomonad history, biology, clinical manifestations, pathogenesis, world distribution, risk factors, diagnosis, and treatment.

Resumo Tritrichomonas foetus é um parasito que foi identificado definitivamente como agente de tricomoníase, caracterizada por diarreia crônica. T. foetus coloniza porções do intestino grosso dos felinos e se manifesta como uma diarreia crônica e recorrente, com muco e sangue, geralmente irresponsiva às drogas comumente usadas no tratamento. O diagnóstico da infecção por tricomonadídeos é feito pela demonstração de trofozoítos no exame direto de fezes frescas, cultura fecal e subsequente exame microscópico ou extração do DNA do parasito na amostra fecal e amplificação, utilizando-se técnicas moleculares. T. foetus é comumente confundido com outros protozoários flagelados, como Giardia duodenalis e Pentatrichomonas hominis. Sem tratamento adequado, a diarreia pode cessar espontaneamente em meses ou anos, porém os gatos podem permanecer portadores do parasito. Esse artigo pretende servir como fonte de informação para pesquisadores e veterinários, revisando os mais importantes aspectos da tricomoníase felina, como histórico, biologia, manifestações clínicas, patogênese, distribuição mundial, fatores de risco, diagnóstico e tratamento.

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BACKGROUND: Tritrichomonas foetus is the etiologic agent of the sexually transmitted disease Bovine Trichomonosis (BT). In Argentina, BT is endemic and represents a relevant health problem that causes reproductive inefficiency in cattle and large economic losses. Metronidazole is the drug of choice in the treatment of BT. Treatment has been associated with a temporary resolution of the clinical signs but is not able to control the disease. In recent years, the apparition of in vivo and in vitro aerobic and anaerobic resistance leading to ineffective treatments has been reported. AIMS: Thus, the aim of the present study was to explore the susceptibility of six different isolates of T. foetus under aerobic (AC) and anaerobic (ANC) conditions. RESULTS AND DISCUSSION: Six isolates of T. foetus were obtained from samples of preputial smegma of bovine origin. Values of minimum lethal concentration and minimum inhibitory concentration were higher than those observed in other works and represent current data in Argentina and provide information to establish new treatment protocols.

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Tritrichomonas foetus is a parasite that has been definitively identified as an agent of trichomonosis, a disease characterized by chronic diarrhea. T. foetus colonizes portions of the feline large intestine, and manifests as chronic and recurrent diarrhea with mucus and fresh blood, which is often unresponsive to common drugs. Diagnosis of a trichomonad infection is made by either the demonstration of the trophozoite on a direct fecal smear, fecal culture and subsequent microscopic examination of the parasite, or extraction of DNA in feces and amplification by the use of molecular tools. T. foetus is commonly misidentified as other flagellate protozoa such as Giardia duodenalis and Pentatrichomonas hominis. Without proper treatment, the diarrhea may resolve spontaneously in months to years, but cats can remain carriers of the parasite. This paper intends to serve as a source of information for investigators and veterinarians, reviewing the most important aspects of feline trichomonosis, such as trichomonad history, biology, clinical manifestations, pathogenesis, world distribution, risk factors, diagnosis, and treatment.(AU)

Tritrichomonas foetus é um parasito que foi identificado definitivamente como agente de tricomoníase, caracterizada por diarreia crônica. T. foetus coloniza porções do intestino grosso dos felinos e se manifesta como uma diarreia crônica e recorrente, com muco e sangue, geralmente irresponsiva às drogas comumente usadas no tratamento. O diagnóstico da infecção por tricomonadídeos é feito pela demonstração de trofozoítos no exame direto de fezes frescas, cultura fecal e subsequente exame microscópico ou extração do DNA do parasito na amostra fecal e amplificação, utilizando-se técnicas moleculares. T. foetus é comumente confundido com outros protozoários flagelados, como Giardia duodenalis e Pentatrichomonas hominis. Sem tratamento adequado, a diarreia pode cessar espontaneamente em meses ou anos, porém os gatos podem permanecer portadores do parasito. Esse artigo pretende servir como fonte de informação para pesquisadores e veterinários, revisando os mais importantes aspectos da tricomoníase felina, como histórico, biologia, manifestações clínicas, patogênese, distribuição mundial, fatores de risco, diagnóstico e tratamento.(AU)

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Bovine genital campylobacteriosis (BGC) and bovine genital trichomonosis (BGT) are globally widespread, predominantly sexually transmitted diseases that mainly affect herds in which natural mating takes place. Bulls serve as good epidemiological indicators for studying the causative agents of these diseases in cattle herds. The objective of the present work was to determine the prevalence of BGC and BGT among bulls sent to slaughterhouses in southern Minas Gerais, Brazil. Samples of preputial smegma from 200 sexually mature, non-castrated bulls of several ancestries were collected in 2013 from four regional slaughterhouses. The polymerase chain reaction (PCR) was employed to detect Tritrichomonas foetus, Campylobacter fetus subsp. fetus, and C. fetus subsp. venerealis. Isolation also was used aiming to diagnosis of BGT. All smegma samples were negative in culture for T. foetus. Molecular tests revealed 8% (16/200) of animals to be positive for T. foetus, 17.5% (35/200) for Campylobacter. fetus subsp. fetus, and 13.5% (27/200) for C. fetus subsp. venerealis. The results of the present study indicate that T. foetus, C. fetus subsp. venerealis, and C. fetus subsp. fetus are present among bulls slaughtered in southern Minas Gerais, and that BGC and BGT occur in this region. These diseases therefore need to be considered during differential diagnosis of reproductive diseases affecting cattle herds in which natural mating is employed.(AU)

A Campilobacteriose Genital Bovina (CGB) e a Tricomonose Genital Bovina (TGB) são doenças de transmissão predominantemente venérea, de ampla disseminação mundial, acometendo principalmente rebanhos submetidos à monta natural. Touros constituem bons indicadores epidemiológicos para a pesquisa destes agentes nos rebanhos. O objetivo deste estudo foi determinar a prevalência da CGB e da TGB em touros encaminhados a abatedouros da região sul de Minas Gerais, Brasil. Amostras de esmegma prepucial de 200 touros (machos não castrados em idade compatível com a monta) de diversas genealogias foram coletadas em quatro abatedouros da região, no ano de 2013. A reação em cadeia da polimerase (PCR) foi utilizada para detecção dos patógenos Tritrichomonas foetus, Campylobacter fetus subsp. fetus e Campylobacter fetus subsp. venerealis. O isolamento também foi utilizado para o diagnóstico da TGB. Todas as amostras analisadas apresentaram resultado negativo na cultura para isolamento de T. foetus. Por meio dos testes moleculares, verificaram-se prevalências de 8% (16/200) de animais positivos para T. foetus; 17,5% (35/200) para C. fetus subsp. fetus e de 13,5% (27/200) para C. fetus subsp. venerealis. Os dados do presente estudo apontaram a presença de T. foetus, C. fetus subsp. venerealis e C. fetus subsp. fetus em touros abatidos na região sul de Minas Gerais, evidenciando a ocorrência da TGB e da CGB nesta região e a necessidade de inclusão destas enfermidades no diagnóstico diferencial das doenças da reprodução em rebanhos bovinos submetidos à monta natural.(AU)

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OBJECTIVES: For over two decades, feline trichomonosis caused by Tritrichomonas foetus has been recognized as a large-bowel protozoan disease of the domestic cat. It has a wide distribution, but no reports exist in the Caribbean. The objectives of this study were to detect the presence of T foetus and its prevalence in the domestic cat on St Kitts, West Indies. METHODS: A cross-sectional study was performed between September 2014 and December 2015. This study recruited 115 feral cats from a trap-neuter-return program and 37 owned cats treated as outpatients at the university veterinary clinic. Fresh feces were inoculated in InPouch culture medium, as per the manufacturer's instructions. In addition, PCR was performed using primers for T foetus. DNA extraction with amplification using primers of a Feliscatus NADH dehydrogenase subunit 6 was used as a housekeeping gene for quality control. RESULTS: Only two owned cats had reported diarrhea in the preceding 6 months. None of the 152 samples were positive on InPouch culture microscopic examination. Only 35/69 feral cat fecal DNA samples were positive for the housekeeping gene, of which none tested PCR positive for T foetus. CONCLUSIONS AND RELEVANCE: T foetus was not detected by culture and PCR in feral cats and owned cats on St Kitts. A high proportion of PCR inhibitors in the DNA samples using a commercial fecal DNA kit can lead to underestimating the prevalence, which should be taken into consideration when a survey on gastrointestinal pathogens depends exclusively on molecular detection.

RESUMO

The objective of this study was to determine the prevalence of Tritrichomonas foetus infection and to evaluate risk factors associated with this infection among cattle in the state of Paraíba in northeastern Brazil. Samples of cervicovaginal mucus from 290 females and smegma from 59 males [beef, 31; mixed aptitude (beef and dairy), 10; and dairy, 18] from 31 farms were collected. Modified Diamond's medium and polymerase chain reaction (PCR) were used for the laboratory diagnosis of T. foetus infection. Univariate analysis and logistic regression were performed to test for potential risk factors in addition to prevalence mapping. No sample was positive for T. foetus in culture, and the prevalence of T. foetus infection using PCR was 3.7% (13/349) [confidence interval (CI) 95%, 2.1%-6.4%]. In total, 19.3% (6/31) of the farms had at least one animal positive for T. foetus. The contact of females with males from other farms [Odds ratio, 5.9; 95% CI, 1.5-22.4; p = 0.009] was identified as a risk factor for T. foetus infection. This study demonstrates that T. foetus infection is prevalent among dairy cows in the state of Paraíba, Brazil. Sexual resting, removal of positive females, and avoiding contact of females with males from other farms are recommended to reduce the risk of infection.

Assuntos

RESUMO

Bovine genital campylobacteriosis (BGC) and bovine genital trichomonosis (BGT) are globally widespread, predominantly sexually transmitted diseases that mainly affect herds in which natural mating takes place. Bulls serve as good epidemiological indicators for studying the causative agents of these diseases in cattle herds. The objective of the present work was to determine the prevalence of BGC and BGT among bulls sent to slaughterhouses in southern Minas Gerais, Brazil. Samples of preputial smegma from 200 sexually mature, non-castrated bulls of several ancestries were collected in 2013 from four regional slaughterhouses. The polymerase chain reaction (PCR) was employed to detect Tritrichomonas foetus, Campylobacter fetus subsp. fetus, and C. fetus subsp. venerealis. Isolation also was used aiming to diagnosis of BGT. All smegma samples were negative in culture for T. foetus. Molecular tests revealed 8% (16/200) of animals to be positive for T. foetus, 17.5% (35/200) for Campylobacter. fetus subsp. fetus, and 13.5% (27/200) for C. fetus subsp. venerealis. The results of the present study indicate that T. foetus, C. fetus subsp. venerealis, and C. fetus subsp. fetus are present among bulls slaughtered in southern Minas Gerais, and that BGC and BGT occur in this region. These diseases therefore need to be considered during differential diagnosis of reproductive diseases affecting cattle herds in which natural mating is employed.

A Campilobacteriose Genital Bovina (CGB) e a Tricomonose Genital Bovina (TGB) são doenças de transmissão predominantemente venérea, de ampla disseminação mundial, acometendo principalmente rebanhos submetidos à monta natural. Touros constituem bons indicadores epidemiológicos para a pesquisa destes agentes nos rebanhos. O objetivo deste estudo foi determinar a prevalência da CGB e da TGB em touros encaminhados a abatedouros da região sul de Minas Gerais, Brasil. Amostras de esmegma prepucial de 200 touros (machos não castrados em idade compatível com a monta) de diversas genealogias foram coletadas em quatro abatedouros da região, no ano de 2013. A reação em cadeia da polimerase (PCR) foi utilizada para detecção dos patógenos Tritrichomonas foetus, Campylobacter fetus subsp. fetus e Campylobacter fetus subsp. venerealis. O isolamento também foi utilizado para o diagnóstico da TGB. Todas as amostras analisadas apresentaram resultado negativo na cultura para isolamento de T. foetus. Por meio dos testes moleculares, verificaram-se prevalências de 8% (16/200) de animais positivos para T. foetus; 17,5% (35/200) para C. fetus subsp. fetus e de 13,5% (27/200) para C. fetus subsp. venerealis. Os dados do presente estudo apontaram a presença de T. foetus, C. fetus subsp. venerealis e C. fetus subsp. fetus em touros abatidos na região sul de Minas Gerais, evidenciando a ocorrência da TGB e da CGB nesta região e a necessidade de inclusão destas enfermidades no diagnóstico diferencial das doenças da reprodução em rebanhos bovinos submetidos à monta natural.

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Proteasomal proteolysis is required for a wide range of cellular processes, including protein quality control, cell cycle progression, cell death and metabolic adaptation to environment changes or stress responses. Proteasome inhibitors are useful compounds for determining the roles of proteasome in eukaryotic cells. Here, we investigated the effects of gliotoxin, a proteasome inhibitor, on the cell growth, replication, ultrastructure, DNA integrity and proteasomal proteolytic activity of the protist parasite Tritrichomonas foetus. The effect of gliotoxin on the transformation of T. foetus to endoflagellar form (EFF), also known as pseudocyst, was investigated. Gliotoxin inhibited the culture growth, arrested cell cycle, and provoked a trichomonacidal effect in a dose-dependent manner. Parasites treated with gliotoxin displayed features typical of cell death, such as membrane blebbing, concentric membrane whorls containing remnants of organelles, intense cytosolic and nuclear vacuolisation, chromatin condensation, DNA fragmentation, cytoplasmic disintegration and plasma membrane disruption. The proteasomal peptidase activity was inhibited by gliotoxin in a dose-dependent manner. Gliotoxin treatment also induced an irreversible EFF transformation in a dose/time-dependent manner. We compared morphological characteristics between gliotoxin- and cold-induced EFF parasites. Our results suggest that gliotoxin could induce EFF transformation by a mechanism distinct from that provoked by cold temperature. This study further contributes to a better understanding of the role of proteasome system in cell cycle, cell death and EFF transformation in T. foetus.

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Tritrichomonas foetus é um protozoário patogênico responsável por doença venérea em bovinos conhecida por tricomonose genital bovina. A tricomonose bovina é uma doença venérea causada pelo protozoário cujo habitat natural é o trato genital. Os protocolos já desenvolvidos para o diagnóstico deste parasito por PCR, apesar de serem eficazes na identificação do DNA genômico alvo, promovem algumas amplificações inespecíficas ou são incapazes de distinguir T. foetus das outras espécies do gênero. O presente trabalho foi desenvolvido com o objetivo de estabelecer e otimizar protocolos de ensaio de PCR e nested-PCR para o diagnóstico específico de T. foetus, empregando-se novos iniciadores, selecionados do alinhamento das seqüências dos genes 18S rRNA, 5,8S rRNA, 28S rRNA e dos espaços transcritos do rDNA (ITS1 e ITS2). Um par de iniciadores foi construído para amplificação gênero-específica de um fragmento de 648 pares de base e outros dois para a obtenção de produtos espécie- específicos de 343 e 429 pb. Nenhuma reação cruzada foi observada frente ao DNA genômico de Bos taurus ou de microrganismos responsáveis por infecções genitais. A sensibilidade dos ensaios de PCR e de nested-PCR apresentados neste estudo permitiu um limiar de detecção de até dois parasitos.

Tritrichomonas foetus is a pathogenic protozoan that causes a venereal disease in cattle known as bovine genital tricomonosis. In spite of the efficacy to recognize the target genomic DNA, the protocols so far developed for the diagnosis of this organism by PCR promote some inespecific amplifications or they are unable to discriminate T. foetus against other species within the genus. The objective of this study was to assess and optimize PCR and nested-PCR assays for the specific diagnosis of T. foetus, using novel primers selected from the alignment of sequences of the genes 18S rRNA, 5.8S rRNA, 28S rRNA and of the internal transcribed spacers of the rDNA (ITS1 and ITS2). A pair of primers was constructed for the genus-specific amplification of a 648 bp fragment and two others to amplify T. foetus species-specific fragments of 343 and 429 bp. No cross amplification was observed against Bos taurus genomic DNA neither against the DNA of usual bovine genital pathogens. Both, single and nested-PCR assays, presented analytical sensitivity to detect at least two T. foetus organisms.

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Tritrichomonas foetus is a pathogenic protozoan that causes a venereal disease in cattle known as bovine genital tricomonosis. In spite of the efficacy to recognize the target genomic DNA, the protocols so far developed for the diagnosis of this organism by PCR promote some inespecific amplifications or they are unable to discriminate T. foetus against other species within the genus. The objective of this study was to assess and optimize PCR and nested-PCR assays for the specific diagnosis of T. foetus, using novel primers selected from the alignment of sequences of the genes 18S rRNA, 5.8S rRNA, 28S rRNA and of the internal transcribed spacers of the rDNA (ITS1 and ITS2). A pair of primers was constructed for the genus-specific amplification of a 648 bp fragment and two others to amplify T. foetus species-specific fragments of 343 and 429 bp. No cross amplification was observed against Bos taurus genomic DNA neither against the DNA of usual bovine genital pathogens. Both, single and nested-PCR assays, presented analytical sensitivity to detect at least two T. foetus organisms.

Tritrichomonas foetus é um protozoário patogênico responsável por doença venérea em bovinos conhecida por tricomonose genital bovina. A tricomonose bovina é uma doença venérea causada pelo protozoário cujo habitat natural é o trato genital. Os protocolos já desenvolvidos para o diagnóstico deste parasito por PCR, apesar de serem eficazes na identificação do DNA genômico alvo, promovem algumas amplificações inespecíficas ou são incapazes de distinguir T. foetus das outras espécies do gênero. O presente trabalho foi desenvolvido com o objetivo de estabelecer e otimizar protocolos de ensaio de PCR e nested-PCR para o diagnóstico específico de T. foetus, empregando-se novos iniciadores, selecionados do alinhamento das seqüências dos genes 18S rRNA, 5,8S rRNA, 28S rRNA e dos espaços transcritos do rDNA (ITS1 e ITS2). Um par de iniciadores foi construído para amplificação gênero-específica de um fragmento de 648 pares de base e outros dois para a obtenção de produtos espécie- específicos de 343 e 429 pb. Nenhuma reação cruzada foi observada frente ao DNA genômico de Bos taurus ou de microrganismos responsáveis por infecções genitais. A sensibilidade dos ensaios de PCR e de nested-PCR apresentados neste estudo permitiu um limiar de detecção de até dois parasitos.