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1.
Vet Res ; 52(1): 129, 2021 Oct 07.
Article in English | MEDLINE | ID: mdl-34620238

ABSTRACT

Bovine trichomonosis is caused by Tritrichomonas foetus. Thirty-three US states have state rules on this disease and render it reportable due to potential huge economic losses to cattle industry. The various rules of different states generally mandate testing and culling T. foetus-positive bulls as well as prohibiting import of T. foetus-positive animals. Wyoming has enforced these rules for over 20 year beginning in 2000. From 2017 to 2019, 3 years in a row, not even one T. foetus-positive bull has been detected throughout the entire state among over ten thousand bulls tested annually. Wyoming is the first US state to achieve total control and eradication of bovine trichomonosis by testing and culling T. foetus-positive bulls.


Subject(s)
Abortion, Veterinary/prevention & control , Cattle Diseases/prevention & control , Protozoan Infections, Animal/parasitology , Tritrichomonas foetus/physiology , Abortion, Veterinary/parasitology , Animals , Cattle , Cattle Diseases/parasitology , Female , Male , Protozoan Infections, Animal/prevention & control , Wyoming
2.
PLoS One ; 16(2): e0246957, 2021.
Article in English | MEDLINE | ID: mdl-33606740

ABSTRACT

The protozoal pathogen Tritrichomonas foetus infects the colon of domestic cats and is a major cause of chronic colitis and diarrhea. Treatment failure is common, but antibiotics may improve clinical signs in a subset of cats, leading researchers to question involvement of the colonic microbiota in disease pathogenesis. Studies performed in women with venereal Trichomonas vaginalis infections have revealed that dysbiosis of host microbiota contributes to pathogenicity with similar findings also found in mice with intestinal Tritrichomonas musculis The aim of this study was to characterize differences in the fecal microbiota of cats with and without naturally occurring T. foetus infection and in a group of kittens prior to and after experimentally induced infection. Archived fecal DNA from cats undergoing testing for T. foetus infection (n = 89) and experimentally infected kittens (n = 4; at pre-, 2 weeks, and 9 weeks post-infection) were analyzed by sequencing of 16S rRNA genes. Amongst the naturally infected population, the genera Megamonas and Helicobacter were significantly increased in prevalence and abundance in cats testing positive for T. foetus infection. In the group of four experimentally infected kittens, fecal samples post-infection had significantly lower abundance of genus Dialister and Megamonas and greater abundance of the class Betaproteobacteria and family Succinivibrionaceae. We hypothesize that T. foetus promotes dysbiosis by competition for fermentable substrates used by these bacteria and that metabolic byproducts may contribute to the pathogenesis of colonic inflammation and diarrhea. Future studies are warranted for the measurement of fecal concentrations of microbial and protozoal metabolites in cats with T. foetus infection for the identification of potential therapeutic targets.


Subject(s)
Dysbiosis/complications , Feces/microbiology , Microbiota , Protozoan Infections/complications , Tritrichomonas foetus/physiology , Animals , Cats , Disease Models, Animal , Mice , Protozoan Infections/microbiology
3.
Cell Microbiol ; 22(11): e13257, 2020 11.
Article in English | MEDLINE | ID: mdl-32858768

ABSTRACT

Extracellular vesicles (EVs) have emerged as a ubiquitous mechanism for transferring information between cells and organisms across all three kingdoms of life. Parasitic unicellular eukaryotes use EVs as vehicles for intercellular communication and host manipulation. Pathogenic protozoans are able to modulate the immune system of the host and establish infection by transferring a wide range of molecules contained in different types of EVs. In addition to effects on the host, EVs are able to transfer virulence factors, drug-resistance genes and differentiation factors between parasites. In this review we cover the current knowledge on EVs from anaerobic or microaerophilic extracellular protozoan parasites, including Trichomonas vaginalis, Tritrichomonas foetus, Giardia intestinalis and Entamoeba histolytica, with a focus on their potential role in the process of infection. The role of EVs in host: parasite communication adds a new level of complexity to our understanding of parasite biology, and may be a key to understand the complexity behind their mechanism of pathogenesis.


Subject(s)
Entamoeba histolytica/physiology , Extracellular Vesicles/metabolism , Giardia lamblia/physiology , Host-Parasite Interactions , Trichomonas/physiology , Anaerobiosis , Animals , Entamoeba histolytica/pathogenicity , Entamoebiasis , Giardia lamblia/pathogenicity , Giardiasis/parasitology , Humans , Protozoan Proteins/metabolism , Trichomonas/pathogenicity , Trichomonas Infections/parasitology , Trichomonas vaginalis/pathogenicity , Trichomonas vaginalis/physiology , Tritrichomonas foetus/pathogenicity , Tritrichomonas foetus/physiology
4.
J Eukaryot Microbiol ; 65(1): 28-37, 2018 01.
Article in English | MEDLINE | ID: mdl-28477402

ABSTRACT

The flagellated protist Tritrichomonas foetus is a parasite that causes bovine trichomonosis, a major sexually transmitted disease in cattle. Cell division has been described as a key player in controlling cell survival in other cells, including parasites but there is no information on the regulation of this process in T. foetus. The regulation of cytokinetic abscission, the final stage of cell division, is mediated by members of the ESCRT (endosomal sorting complex required for transport) machinery. VPS32 is a subunit within the ESCRTIII complex and here, we report that TfVPS32 is localized on cytoplasmic vesicles and a redistribution of the protein to the midbody is observed during the cellular division. In concordance with its localization, deletion of TfVPS32 C-terminal alpha helices (α5 helix and/or α4-5 helix) leads to abnormal T. foetus growth, an increase in the percentage of multinucleated parasites and cell cycle arrest at G2/M phase. Together, these results indicate a role of this protein in controlling normal cell division.


Subject(s)
Cell Division/genetics , Protozoan Proteins/genetics , Tritrichomonas foetus/physiology , Cytokinesis/genetics , Protozoan Proteins/metabolism , Tritrichomonas foetus/genetics
5.
Exp Parasitol ; 173: 18-28, 2017 Feb.
Article in English | MEDLINE | ID: mdl-27956086

ABSTRACT

Tritrichomonas foetus infects the bovine urogenital tract, causing bovine trichomoniasis. Significant economic losses may occur due to infertility and abortion among cattle. Trichomonas vaginalis is the causative agent of trichomoniasis; the most common but overlooked non-viral sexually transmitted disease worldwide. Human and bovine trichomoniasis present treatment restrictions and efforts to identify new alternatives are essential. The present study evaluated the anti-trichomonads activities of seven fractions from northwest endemic plant Manilkara rufula. Flavonoids and condensed tannins were identified from these fractions by LC-DAD-MS/MS and MALDI-MS/MS. Altogether, the results demonstrated for the first time the structural description of tannins from leaves of M. rufula and the relation of these compounds with anti-T. vaginalis and anti-T. foetus activities. Overall, this report reveals the potential of M. rufula fractions against both parasites and shows new alternatives to treat the infection caused by trichomonads.


Subject(s)
Antitrichomonal Agents/pharmacology , Flavonoids/pharmacology , Manilkara/chemistry , Plant Extracts/pharmacology , Tannins/pharmacology , Trichomonas vaginalis/drug effects , Tritrichomonas foetus/drug effects , Antitrichomonal Agents/chemistry , Antitrichomonal Agents/isolation & purification , Brazil , Cell Line , Cell Survival , Chromatography, Liquid , Epithelial Cells/drug effects , Flavonoids/chemistry , Flavonoids/isolation & purification , HeLa Cells , Humans , Molecular Structure , Plant Extracts/chemistry , Plant Extracts/isolation & purification , Plant Leaves/chemistry , Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization , Tannins/chemistry , Tannins/isolation & purification , Trichomonas vaginalis/physiology , Tritrichomonas foetus/physiology
6.
Vet Parasitol ; 221: 118-22, 2016 May 15.
Article in English | MEDLINE | ID: mdl-27084482

ABSTRACT

Tritrichomonas foetus is a protozoan parasite that has been recently identified as a causative agent of chronic diarrhea in domestic cats. Transmission of infection occurs by the fecal-oral route through direct contact among animals. Consequently, feline trichomonosis (FT) is more likely to be present in multi-cat environments. The objective of this work was to study the presence of T. foetus and some associated risk factors in cats from densely housed origins and with a reported history of chronic diarrhea. Animals enrolled in this study were family cats (n=15) acquired from pet shops, shelters or breeding centers and cattery cats belonging to one breeding center (n=28) and two cat shelters (A and B, n=25 each). In the catteries, a follow-up analysis for a period of up to 2 months was also performed to determine the parasite shedding pattern in feces and the incidence of infection. Fecal samples were analyzed using in vitro culture and a PCR technique. T. foetus was detected in a total of 38.7% (36/93) of the cats with chronic diarrhea. Parasite infection was similarly detected in family cats and cattery animals (40% versus 38.4%). In the catteries, the parasite was detected in 50%, 44% and 20% of the animals from the breeding center and shelters A and B, respectively. The follow-up analysis showed that 58.3% of infected cats intermittently shed trophozoites in their feces, with an incidence of 23.1%. Investigation of potential risk factors showed that cats ≤1 year old were more likely to be infected than older cats (57.1% versus 27.3%; P<0.05). No significant differences were found when sex and breed factors were studied. These results confirm the importance of FT as a cause of chronic diarrhea in cats and highlight the relevance of close contact conditions for T. foetus transmission.


Subject(s)
Cat Diseases/epidemiology , Diarrhea/veterinary , Housing, Animal/standards , Protozoan Infections, Animal/epidemiology , Tritrichomonas foetus/physiology , Age Factors , Animals , Cats , Diarrhea/etiology , Feces/parasitology , Incidence , Population Density , Protozoan Infections, Animal/complications , Risk Factors , Spain
7.
Theriogenology ; 85(5): 781-791, 2016 Mar 15.
Article in English | MEDLINE | ID: mdl-26679515

ABSTRACT

The objective is to discuss sexually transmitted diseases caused by Tritrichomonas foetus (T foetus) and Campylobacter fetus (C fetus) subsp. venerealis, with a focus on prevalence, pathogenesis, and diagnosis in cows and bulls. Diagnosis and control are problematic because these diseases cause severe reproductive losses in cows, but in bulls are clinically asymptomatic, which allows the disease to flourish, especially in the absence of legislated control programs. We review research regarding prophylactic systemic immunization of bulls and cows with antigens of T foetus and C fetus venerealis and their efficacy in preventing or clearing preexisting infections in the genital tract. Current diagnostic methods of C fetus venerealis and T foetus (microbial culture and PCR) should be improved. Review of the latest advances in bovine trichomoniasis and campylobacteriosis should promote knowledge and provide an impetus to pursue further efforts to control bovine sexually transmitted diseases.


Subject(s)
Campylobacter Infections/physiopathology , Campylobacter fetus/physiology , Cattle Diseases/physiopathology , Protozoan Infections, Animal/physiopathology , Reproduction/physiology , Sexually Transmitted Diseases/physiopathology , Tritrichomonas foetus/physiology , Animals , Campylobacter Infections/epidemiology , Campylobacter Infections/therapy , Campylobacter Infections/veterinary , Campylobacter fetus/pathogenicity , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/epidemiology , Cattle Diseases/therapy , Female , Male , Mass Vaccination/veterinary , Protozoan Infections, Animal/diagnosis , Protozoan Infections, Animal/epidemiology , Protozoan Infections, Animal/therapy , Sexually Transmitted Diseases/epidemiology , Sexually Transmitted Diseases/therapy , Sexually Transmitted Diseases/veterinary , Tritrichomonas foetus/pathogenicity
8.
J Vet Med Sci ; 78(4): 633-40, 2016 May 03.
Article in English | MEDLINE | ID: mdl-26685985

ABSTRACT

Tritrichomonas species flagellates (IMC strain) were isolated from the biliary tract of an individual who had developed cholecystitis as a complication of acquired agammaglobulinemia. Sequence analysis of Tritrichomonas sp. (IMC clone 2 (cl2)) was performed for several genetic regions including the ITS1-5.8S rDNA-ITS2 region, the cysteine protease (CP)-1, CP-2 and CP-4 to CP-9 genes, and the cytosolic malate dehydrogenase 1 gene. In addition to comparison of the variable-length DNA repeats in the isolate clone with those in T. foetus (Inui cl2) and the T. mobilensis (U.S.A.: M776 cl2) reference strains, this analysis showed that the Tritrichomonas sp. (IMC cl2) was T. foetus (cattle/swine genotype). Injection of T. foetus (IMC cl2) directly into the livers of CBA mice resulted in liver abscess formation on Day 7. Moreover, inoculation via orogastric intubation caused infection in the cecum on Day 5 in CBA mice co-infected with Entamoeba histolytica (HM-1: IMSS cl6). T. foetus (IMC cl2) was able to grow in YI-S medium for over 20 days, even at 5°C. These results indicate that the T. foetus isolate is able to survive in the feces and edible organ meat of the definitive host for a prolonged period of time, and it is possible that the parasite could infect humans.


Subject(s)
Protozoan Infections/parasitology , Tritrichomonas foetus/isolation & purification , Tritrichomonas foetus/physiology , Zoonoses/parasitology , Adult , Agammaglobulinemia/complications , Animals , Cholecystitis/etiology , Colitis/parasitology , Humans , Male , Mice , Mice, Inbred CBA , Molecular Typing , Protozoan Infections/transmission , Tritrichomonas foetus/classification , Tritrichomonas foetus/genetics
9.
Exp Parasitol ; 157: 68-77, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26160677

ABSTRACT

The ability for protozoan parasites to tolerate pH fluctuations within their niche is critical for the establishment of infection and require the parasite to be capable of adapting to a distinct pH range. We used two host adapted Tritrichomonas foetus isolates, capable of infecting either the digestive tract (pH 5.3-6.6) of feline hosts or the reproductive tract (pH 7.4-7.8) of bovine hosts to address their adaptability to changing pH. Using flow cytometry, we investigated the pH tolerance of the bovine and feline T. foetus isolates over a range of physiologically relevant pH in vitro. Following exposure to mild acid stress (pH 6), the bovine T. foetus isolates showed a significant decrease in cell viability and increased cytoplasmic granularity (p-value < 0.003, p-value < 0.0002) compared to pH 7 and 8 (p-value > 0.7). In contrast, the feline genotype displayed an enhanced capacity to maintain cell morphology and viability (p-value > 0.05). Microscopic assessment revealed that following exposure to a weak acidic stress (pH 6), the bovine T. foetus transformed into rounded parasites with extended cell volumes and displays a decrease in viability. The higher tolerance for acidic extracellular environment of the feline isolate compared to the bovine isolate suggests that pH could be a critical factor in regulating T. foetus infections and host-specificity.


Subject(s)
Cat Diseases/parasitology , Cattle Diseases/parasitology , Gastrointestinal Tract/parasitology , Protozoan Infections, Animal/parasitology , Tritrichomonas foetus/physiology , Urogenital System/parasitology , Adaptation, Physiological , Animals , Cats , Cattle , Flow Cytometry/veterinary , Fluorescent Dyes , Gastrointestinal Tract/chemistry , Genotype , Host-Parasite Interactions , Hydrogen-Ion Concentration , Microscopy, Electron, Transmission/veterinary , Tritrichomonas foetus/isolation & purification , Tritrichomonas foetus/ultrastructure , Urogenital System/chemistry
10.
Vet Res ; 46: 35, 2015 Mar 19.
Article in English | MEDLINE | ID: mdl-25880025

ABSTRACT

Tritrichomonas foetus is a very intriguing trichomonad protozoan with respect to its varied choice of residence in the different host species. It is an obligate parasite of the reproductive and the gastrointestinal tract of bovine and feline host respectively, leading to trichomonosis. Bovine trichomonosis is a sexually transmitted disease whereas feline trichomonosis is a disease with a purported fecal-oral route of spread. Further, the trichomonad is a commensal in the nasal passages, stomach, cecum and colon of swine host. Advances have been exponential in understanding the trichomonad biology and specifically feline trichomonosis since late 1990s and early 2000s when T. foetus was soundly determined to be a causative agent of chronic diarrhea in the domestic cat. It is a challenging task, even for a skilled investigator not to mention the busy clinical veterinarian, to keep up with the vast volume of information. Here we comprehensively reviewed the trichomonad biology, clinical manifestations, pathogenesis, host immunity, world map of distribution, risk factors, diagnosis and treatment. Risk factors associated with T. foetus-positive status in the domestic cat include young age, purebred, history of diarrhea, co-infections with other enteral pathogens. In addition, molecular similarity of bovine and feline isolates of T. foetus in DNA sequence was concisely discussed. The data presented serve as an information source for veterinarians, and investigators who are interested in biology of T. foetus and feline trichomonosis.


Subject(s)
Cat Diseases/parasitology , Diarrhea/veterinary , Protozoan Infections, Animal/parasitology , Tritrichomonas foetus/physiology , Animals , Cat Diseases/diagnosis , Cat Diseases/epidemiology , Cat Diseases/prevention & control , Cats , Diarrhea/epidemiology , Diarrhea/parasitology , Diarrhea/prevention & control , Protozoan Infections, Animal/diagnosis , Protozoan Infections, Animal/epidemiology , Protozoan Infections, Animal/prevention & control , Risk Factors , Tritrichomonas foetus/genetics
11.
Protist ; 165(3): 293-304, 2014 May.
Article in English | MEDLINE | ID: mdl-24742927

ABSTRACT

Tritrichomonas foetus is a serious veterinary parasite that causes bovine trichomoniasis, a sexually transmitted disease that results in reproductive failure and considerable economic losses in areas that practice natural breeding. T. foetus is an extracellular parasite, which initially adheres to and infects the urogenital tract using a diverse array of surface glycoconjugates, including adhesins and extracellular matrix-binding molecules. However, the cellular mechanisms by which T. foetus colonizes mucosal surfaces and causes tissue damage are not well defined. Several studies have demonstrated the involvement of pathogen or host lipid rafts in cellular events that occur during pathogenesis, including adhesion, invasion and evasion of the immune response. In this study, we demonstrate that detergent-resistant membranes are present in the plasma membrane of T. foetus. We further demonstrate that microdomains are cholesterol-enriched and contain ganglioside GM1-like molecules. In addition, we demonstrate that lipid microdomains do not participate in T. foetus adhesion to host cells. However, the use of agents that disrupt and disorganize the plasma membrane indicated the involvement of the T. foetus lipid microdomains, in cell division and in the formation of endoflagellar forms. Our results suggest that trophozoites and endoflagellar forms present a different plasma membrane organization.


Subject(s)
Detergents/pharmacology , Membrane Microdomains/drug effects , Tritrichomonas foetus/drug effects , Animals , Cattle , Cattle Diseases/parasitology , Cell Adhesion/drug effects , Cell Membrane/chemistry , Cell Membrane/drug effects , Cell Membrane/metabolism , Gangliosides/metabolism , Membrane Microdomains/chemistry , Membrane Microdomains/metabolism , Mucous Membrane/parasitology , Protozoan Infections, Animal/parasitology , Tritrichomonas foetus/chemistry , Tritrichomonas foetus/physiology
12.
Vet Parasitol ; 200(3-4): 289-94, 2014 Mar 01.
Article in English | MEDLINE | ID: mdl-24447668

ABSTRACT

Switzerland is officially free from bovine Tritrichomonas foetus. While bulls used for artificial insemination (AI) are routinely examined for this pathogen, bulls engaged in natural mating, as well as aborted fetuses, are only very sporadically investigated, indicating that the disease awareness for bovine tritrichomoniasis is low. Natural mating in cattle is becoming increasingly popular in Switzerland. Accordingly, a re-introduction/re-occurrence of T. foetus in cattle seems possible either via resurgence from a yet unknown bovine reservoir, or via importation of infected cattle. The low disease awareness for bovine tritrichomoniasis might favor an unnoticed re-establishment of T. foetus in the Swiss cattle population. The aim of our study was thus to search for the parasite, and if found, to assess the prevalence of bovine T. foetus in Switzerland. We included (1) bulls over two years of age used in natural mating and sent to slaughter, (2) bulls used for natural service in herds with or without fertility problems and (3) aborted fetuses. Furthermore, the routinely examined bulls used for AI (4) were included in this study. In total, 1362 preputial samples from bulls and 60 abomasal fluid samples of aborted fetuses were analyzed for the presence of T. foetus by both in vitro cultivation and molecular analyses. The parasite could not be detected in any of the samples, indicating that the maximal prevalence possibly missed was about 0.3% (95% confidence). Interestingly, in preputial samples of three bulls of category 1, apathogenic Tetratrichomonas sp. was identified, documenting a proof-of-principle for the methodology used in this study.


Subject(s)
Cattle Diseases/epidemiology , Protozoan Infections, Animal/epidemiology , Tritrichomonas foetus/physiology , Abomasum/parasitology , Aborted Fetus/parasitology , Animals , Breeding , Cattle , Gastrointestinal Contents/parasitology , Male , Penis/parasitology , Prevalence , Switzerland/epidemiology , Trichomonadida/genetics , Trichomonadida/isolation & purification
13.
J R Soc Interface ; 11(93): 20131149, 2014 Apr 06.
Article in English | MEDLINE | ID: mdl-24478286

ABSTRACT

In this work, a high-speed imaging platform and a resistive force theory (RFT) based model were applied to investigate multi-flagellated propulsion, using Tritrichomonas foetus as an example. We discovered that T. foetus has distinct flagellar beating motions for linear swimming and turning, similar to the 'run and tumble' strategies observed in bacteria and Chlamydomonas. Quantitative analysis of the motion of each flagellum was achieved by determining the average flagella beat motion for both linear swimming and turning, and using the velocity of the flagella as inputs into the RFT model. The experimental approach was used to calculate the curvature along the length of the flagella throughout each stroke. It was found that the curvatures of the anterior flagella do not decrease monotonically along their lengths, confirming the ciliary waveform of these flagella. Further, the stiffness of the flagella was experimentally measured using nanoindentation, allowing for calculation of the flexural rigidity of T. foetus's flagella, 1.55×10(-21) N m(2). Finally, using the RFT model, it was discovered that the propulsive force of T. foetus was similar to that of sperm and Chlamydomonas, indicating that multi-flagellated propulsion does not necessarily contribute to greater thrust generation, and may have evolved for greater manoeuvrability or sensing. The results from this study have demonstrated the highly coordinated nature of multi-flagellated propulsion and have provided significant insights into the biology of T. foetus.


Subject(s)
Flagella/physiology , Models, Biological , Tritrichomonas foetus/physiology , Movement/physiology
14.
Vet Microbiol ; 164(1-2): 171-6, 2013 May 31.
Article in English | MEDLINE | ID: mdl-23434012

ABSTRACT

Research on microbiota in cattle tick and the evaluation of its activity against other microorganisms can contribute to identify new molecules potentially useful to control infections caused by bacteria and protozoa. Biofilms pose increasing problems worldwide, mainly due to their resistance to antimicrobial therapies and host immune response. In this study we investigate the ability Rhipicephalus (Boophilus) microplus-associated bacteria may exhibit to produce anti-biofilm and trichomonicidal compounds. Gut, ovary, salivary glands, and Gené organ were collected from engorged R. microplus female. Homogenates of each tissue were inoculated onto 15 distinct culture media. Anti-biofilm and trichomonicidal activities were analyzed by culturing each bacterium isolated in a liquid medium. Results showed that R. microplus cattle tick microflora varies for different tissues. Bacteria belonging to different genera (Aeromonas, Bacillus, Brevibacillus, Castelaniella, Comamonas, Kocuria, and Microbacterium) were identified. Interestingly, all bacterial species found displayed pronounced activity against Staphylococcus epidermidis and Pseudomonas aeruginosa biofilms, and also against the cattle pathogen Tritrichomonas foetus, confirming the hypothesis that cattle tick could be a source of bacteria active against pathogens. This is the first study showing that bacteria isolated from a tick exert anti-biofilm and trichomonicidal activities.


Subject(s)
Antibiosis , Bacteria/chemistry , Cattle/parasitology , Microbiota , Rhipicephalus/microbiology , Tritrichomonas foetus/physiology , Animals , Bacteria/classification , Bacteria/isolation & purification , Biofilms , Female , Pseudomonas aeruginosa/physiology , Staphylococcus aureus/physiology
15.
Vet Parasitol ; 192(1-3): 75-82, 2013 Feb 18.
Article in English | MEDLINE | ID: mdl-23182300

ABSTRACT

Tritrichomonas foetus (TF) is a protozoan that infects the feline ileum and colon resulting in chronic diarrhea. Up to 30% of young purebred cats are infected with TF and the infection is recognized as pandemic. Only a single drug, characterized by a narrow margin of safety and emerging development of resistance, is effective for treatment. While the venereal pathogenicity of bovine TF is attributed to adherence to uterovaginal epithelium, the pathogenesis of diarrhea in feline TF infection is unknown. The aim of this study was to establish an in vitro model of feline TF adhesion to intestinal epithelium. Confluent monolayers of porcine intestinal epithelial cells (IPEC-J2) were infected with axenic cultures of feline TF that were labeled with [(3)H] thymidine or CFSE and harvested at log-phase. The effect of multiplicity and duration of infection, viability of TF, binding competition, formalin fixation and cytoskeletal inhibitors on adherence of feline TF to IPEC-J2 monolayers was quantified by liquid scintillation counting and immunofluorescence. [(3)H] thymidine and CFSE-labeled TF reproducibly adhered to IPEC-J2 monolayers. Clinical isolates of feline TF adhered to the intestinal epithelium in significantly greater numbers than Pentatrichomonas hominis, the latter of which is a presumably nonpathogenic trichomonad. Adhesion of TF required viable trophozoites but was independent of cytoskeletal activity. Based on saturation and competition binding experiments, adherence of feline TF to the epithelium occurred via specific receptor-ligand interactions. The developed model provides a valuable resource for assessing pathogenic mechanisms of feline TF and developing novel pharmacologic therapies for blocking the adhesion of feline TF to the intestinal epithelium.


Subject(s)
Cat Diseases/parasitology , Intestinal Mucosa/parasitology , Protozoan Infections, Animal/parasitology , Tritrichomonas foetus/physiology , Animals , Cats , Cell Adhesion , Cell Line , Cell Proliferation , Diarrhea/parasitology , Diarrhea/veterinary , Epithelial Cells/parasitology , Ligands , Microscopy, Electron, Scanning , Models, Biological , Swine , Trichomonadida/metabolism , Trichomonadida/physiology , Tritium , Tritrichomonas foetus/metabolism , Trophozoites
16.
Mem Inst Oswaldo Cruz ; 107(6): 720-7, 2012 Sep.
Article in English | MEDLINE | ID: mdl-22990959

ABSTRACT

Trichomonas vaginalis and Tritrichomonas foetus are parasitic, flagellated protists that inhabit the urogenital tract of humans and bovines, respectively. T. vaginalis causes the most prevalent non-viral sexually transmitted disease worldwide and has been associated with an increased risk for human immunodeficiency virus-1 infection in humans. Infections by T. foetus cause significant losses to the beef industry worldwide due to infertility and spontaneous abortion in cows. Several studies have shown a close association between trichomonads and the epithelium of the urogenital tract. However, little is known concerning the interaction of trichomonads with cells from deeper tissues, such as fibroblasts and muscle cells. Published parasite-host cell interaction studies have reported contradictory results regarding the ability of T. foetus and T. vaginalis to interact with and damage cells of different tissues. In this study, parasite-host cell interactions were examined by culturing primary human fibroblasts obtained from abdominal biopsies performed during plastic surgeries with trichomonads. In addition, mouse 3T3 fibroblasts, primary chick embryo myogenic cells and L6 muscle cells were also used as models of target cells. The parasite-host cell cultures were processed for scanning and transmission electron microscopy and were tested for cell viability and cell death. JC-1 staining, which measures mitochondrial membrane potential, was used to determine whether the parasites induced target cell damage. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling staining was used as an indicator of chromatin damage. The colorimetric crystal violet assay was performed to ana-lyse the cytotoxicity induced by the parasite. The results showed that T. foetus and T. vaginalis adhered to and were cytotoxic to both fibroblasts and muscle cells, indicating that trichomonas infection of the connective and muscle tissues is likely to occur; such infections could cause serious risks to the infected host.


Subject(s)
Cell Adhesion/physiology , Fibroblasts/parasitology , Host-Parasite Interactions/physiology , Muscle Cells/parasitology , Trichomonas vaginalis/physiology , Tritrichomonas foetus/physiology , Animals , Chick Embryo , Humans , In Situ Nick-End Labeling , Mice
17.
Mem. Inst. Oswaldo Cruz ; 107(6): 720-727, set. 2012. ilus, graf
Article in English | LILACS | ID: lil-649485

ABSTRACT

Trichomonas vaginalis and Tritrichomonas foetus are parasitic, flagellated protists that inhabit the urogenital tract of humans and bovines, respectively. T. vaginalis causes the most prevalent non-viral sexually transmitted disease worldwide and has been associated with an increased risk for human immunodeficiency virus-1 infection in humans. Infections by T. foetus cause significant losses to the beef industry worldwide due to infertility and spontaneous abortion in cows. Several studies have shown a close association between trichomonads and the epithelium of the urogenital tract. However, little is known concerning the interaction of trichomonads with cells from deeper tissues, such as fibroblasts and muscle cells. Published parasite-host cell interaction studies have reported contradictory results regarding the ability of T. foetus and T. vaginalis to interact with and damage cells of different tissues. In this study, parasite-host cell interactions were examined by culturing primary human fibroblasts obtained from abdominal biopsies performed during plastic surgeries with trichomonads. In addition, mouse 3T3 fibroblasts, primary chick embryo myogenic cells and L6 muscle cells were also used as models of target cells. The parasite-host cell cultures were processed for scanning and transmission electron microscopy and were tested for cell viability and cell death. JC-1 staining, which measures mitochondrial membrane potential, was used to determine whether the parasites induced target cell damage. Terminal deoxynucleotidyltransferase-mediated dUTP nick end labelling staining was used as an indicator of chromatin damage. The colorimetric crystal violet assay was performed to ana-lyse the cytotoxicity induced by the parasite. The results showed that T. foetus and T. vaginalis adhered to and were cytotoxic to both fibroblasts and muscle cells, indicating that trichomonas infection of the connective and muscle tissues is likely to occur; such infections could cause serious risks to the infected host.


Subject(s)
Animals , Chick Embryo , Humans , Mice , Cell Adhesion/physiology , Fibroblasts/parasitology , Host-Parasite Interactions/physiology , Muscle Cells/parasitology , Trichomonas vaginalis/physiology , Tritrichomonas foetus/physiology , In Situ Nick-End Labeling
18.
Vet Parasitol ; 185(2-4): 279-81, 2012 Apr 30.
Article in English | MEDLINE | ID: mdl-22100399

ABSTRACT

Feline intestinal trichomoniasis caused by Tritrichomonas foetus is associated with large bowel diarrhea in cats from many parts of the world. It has long been recognized as an economically important sexually transmitted disease that causes early abortion in cattle. Isolates of T. foetus from cattle are infectious for the large intestine of cats and isolates of T. foetus from cats are infectious for the reproductive system of cattle. The parasite is maintained by fecal-oral transmission in cats. The present study was conducted to examine the survival of a feline isolate of T. foetus, AUTf-12, under various conditions that are relevant to fecal-oral transmission in cats. Trophozoites were grown in TYM medium and then exposed to water, cat urine, dry cat food, canned cat food, clumping cat litter, or filter paper for various lengths of time and then re-cultured in TYM medium. Trophozoites survived exposure to distilled or tap water for 30 but not 60 min, while they survived for at least 180 min in urine. Trophozoites survived for 30 min on dry cat food but survived for 120-180 min in canned cat food. No survival of trophozoites was observed on cat litter but trophozoites survived for 15 min when placed on filter paper. Our results indicate that T. foetus can survive and be potentially infectious in water, urine, dry cat food and canned cat food.


Subject(s)
Animal Feed/parasitology , Cat Diseases/parasitology , Protozoan Infections, Animal/parasitology , Tritrichomonas foetus/classification , Tritrichomonas foetus/physiology , Animals , Cat Diseases/urine , Cats , Male , Tritrichomonas foetus/isolation & purification , Trophozoites/physiology
19.
J Am Vet Med Assoc ; 239(12): 1589-93, 2011 Dec 15.
Article in English | MEDLINE | ID: mdl-22129124

ABSTRACT

OBJECTIVE: To evaluate effects of high incubation temperatures on results of protozoal culture and real-time PCR testing for Tritrichomonas foetus inoculated in a commercially available self-contained culture media system. DESIGN: In vitro experimental study. SAMPLE: 2 strains of T foetus (1 field isolate from the University of California-Davis and 1 field isolate from the Texas Veterinary Medical Diagnostic Laboratory). PROCEDURES: 2 sets of 36 dual-chamber media pouches were inoculated with T foetus (36 sample pouches/strain) and incubated at temperatures of 37.0°C (98.6°F), 46.1°C (115.0°F), or 54.4°C (130.0°F) for 1, 3, 6, or 24 hours. Six uninoculated media samples in pouches stored at 37.0°C for the entire treatment period were used as negative controls. Pouches were removed from incubators and stored at 22.2°C (72.0°F) until all treatments were complete. Samples were submitted to a diagnostic laboratory for protozoal culture and real-time PCR testing. RESULTS: T foetus was detectable microscopically in inoculated pouches incubated at 37.0°C regardless of exposure time, whereas those incubated at 46.1°C yielded T foetus after 1 and 3 hours only, and those incubated at 54.4°C yielded T foetus after 1 hour only. Testing via real-time PCR assay yielded positive results for all inoculated media samples and negative results for all uninoculated control samples. CONCLUSIONS AND CLINICAL RELEVANCE: Samples collected into the self-contained culture media system for T foetus testing via culture alone should be protected from high temperatures. Realtime PCR amplification may be a more reliable method for identification of the organism if storage and transport temperatures cannot be controlled.


Subject(s)
Culture Media , Hot Temperature , Real-Time Polymerase Chain Reaction/veterinary , Tritrichomonas foetus/physiology , Animals , Cattle , Cattle Diseases/diagnosis , Cattle Diseases/parasitology , Protozoan Infections, Animal/diagnosis
20.
Mem Inst Oswaldo Cruz ; 106(6): 701-4, 2011 Sep.
Article in English | MEDLINE | ID: mdl-22012224

ABSTRACT

Trichomonas vaginalis and Tritrichomonas foetus are human and bovine parasites, respectively, that provoke the sexually transmitted disease trichomoniasis. These extracellular parasites adhere to the host epithelial cell surface. Although mucinases and proteases have been described as important proteins for parasite adhesion to epithelial cells, no studies have examined the role of the keratin molecules that cornify the vaginal epithelium. Here, we investigated the interaction of T. vaginalis and T. foetus with human keratin in vitro; additionally, adherence assays were performed in cattle with T. foetus to elucidate whether trichomonads were able to interact with keratin in vivo. We demonstrated that both T. vaginalisand T. foetusinteracted directly with keratin. Additionally, the trichomonads ingested and digested keratin, shedding new light on the Trichomonas infection process.


Subject(s)
Epithelial Cells/parasitology , Keratins/physiology , Trichomonas vaginalis/physiology , Tritrichomonas foetus/physiology , Animals , Cattle , Female , Host-Pathogen Interactions , Humans , Microscopy, Electron , Microscopy, Fluorescence , Trichomonas vaginalis/ultrastructure , Tritrichomonas foetus/ultrastructure , Vagina/parasitology
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