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      Prevalence, risk factors of infection and molecular characterization of trichomonads in puppies from French breeding kennels

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          Abstract

          The trichomonad species Tritrichomonas fetus and Pentatrichomonas hominis were recently identified in the feces of dogs with diarrhea. However the prevalence and pathogenicity of these parasites in the canine population still remained poorly resolved. Therefore the aim of the present study was (1) to determine the prevalence of trichomonads infecting puppies living in French breeding kennels, (2) to confirm the predominance of P. hominis in dogs, (3) to investigate the genetic diversity of P. hominis isolates identified in the French canine population and (4) to evaluate the risk factors for infection by P. hominis and the influence of the parasite on feces consistency. A total of 215 both diarrheic and non-diarrheic puppies from 25 French breeding kennels were included in this epidemiological survey. Fecal samples from each puppy were examined for 6 gastrointestinal pathogens: parvovirus type 2 (CPV2), coronavirus, Toxocara canis, Cystoisospora ohioensis-complex, Cystoisospora canis, and Giardia intestinalis. A part of each collected stool was also tested for the presence of motile trichomonads by microscopy after culturing. The prevalence of trichomonad infection was 15.8% (34/215) among puppies and 20% (5/25) among breeding kennels. DNA from 26 of the 34 positive samples was successfully amplified using a trichomonad-specific primer pair. Analysis of the sequences of PCR products indicated that P. hominis was the only trichomonad infecting the canine population. All the puppies infected with P. hominis belonged to large breed dogs. Moreover, puppies from large breeding kennels, excreting a high level of G. intestinalis and/or excreting a high level of C. canis oocysts showed a higher probability of being positive for P. hominis infection. Univariate analysis also revealed an increased risk for P. hominis infection in puppies with abnormal feces. However, in a multivariate analysis, CPV2 was the only gastrointestinal pathogen associated with abnormal feces. Since enteropathogens were commonly found in dogs infected by P. hominis, the pathogenic potential of this trichomonad species remained uncertain and has to be further evaluated by experimental infection studies.

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          Critical taxonomic revision of Parabasalids with description of one new genus and three new species.

          We propose a new classification of Parabasalia which is congruent with both ultrastructural and molecular-phylogenetic studies. We identify six main parabasalid lineages and give them the rank of class: Hypotrichomonadea, Trichomonadea, Tritrichomonadea, Cristamonadea, Trichonymphea, and Spirotrichonymphea. Trichomonadea is characterized by a single mastigont and by the absence of both a comb-like structure and an infrakinetosomal body. Most representatives also possess a lamelliform undulating membrane. Trichomonadea is divided into two monophyletic orders, Trichomonadida (family Trichomonadidae; with a B-type costa) and Honigbergiellida (families Honigbergiellidae, Hexamastigidae and Tricercomitidae; without a costa). The class Tritrichomonadea, with a single order Tritrichomonadida, is ancestrally characterized by a single mastigont with four flagella, and both a comb-like structure and an infrakinetosomal body. The morphologically most complex representatives (family Tritrichomonadidae) possess in addition a rail-type undulating membrane, an A-type costa, and a suprakinetosomal body. These last three characters are absent in families Monocercomonadidae and Simplicimonadidae. The remaining tritrichomonadids, Dientamoebidae, have undergone reductive evolution. Cristamonads (Cristamonadea) are morphologically derived from tritrichomonads. Because we are unable to determine morphologically homogenous monophyletic lineages within cristamonads, we classify all cristamonads into a single family, Lophomonadidae. Hypotrichomonadea, comprising the genera Trichomitus and Hypotrichomonas, resembles Tritrichomonadea by an A-type costa, and by the presence of a comb-like structure in the mastigont. However, they do not possess an infrakinetosomal body, and are not specifically related to Tritrichomonadea in molecular-phylogenetic analyses. Moreover, unlike Tritrichomonadea, Hypotrichomonadea possesses a lamelliform undulating membrane. The remaining parabasalids are of complex morphology and belong to the classes Trichonymphea and Spirotrichonymphea. A new parabasalid genus, Simplicimonas (Tritrichomonadea), and three new species, Tetratrichomonas undula, Hexamastix coercens and Simplicimonas similis, are described. 2009 Elsevier GmbH. All rights reserved.
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            Prevalence of and risk factors for feline Tritrichomonas foetus and giardia infection.

            Data were gathered for 117 cats from 89 catteries at an international cat show to examine prevalence and risk factors for feline Tritrichomonas foetus and Giardia infection. Prevalence of T. foetus was 31% among cats (36 out of 117) and catteries (28 out of 89) based on results of fecal smear examination (5 out of 36), fecal culture in modified Diamond's medium (9 out of 36), fecal culture in In Pouch TF medium (20 out of 36), or PCR amplification of the ribosomal RNA gene from feces with T. foetus-specific primers (34 out of 36). Catteries in which T. foetus was identified were more likely to have had a recent history of diarrhea, historical diagnosis of coccidia infection in adult cats, and a decreased number of square feet of facility per cat. Evidence did not exist for the ongoing transmission of T. foetus by water, food, or contact with other species.
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              Experimental infection of cats with Tritrichomonas foetus.

              To determine whether infection with Tritrichomonas foetus causes diarrhea in specific-pathogen-free or Cryptosporidium coinfected cats. 4 cats with subclinical cryptosporidiosis (group 1) and 4 specific-pathogen-free cats (group 2). Cats were infected orogastrically with an axenic culture of T. foetus isolated from a kitten with diarrhea. Direct microscopy and protozoal culture of feces, fecal character, serial colonic mucosal biopsy specimens, and response to treatment with nitazoxanide (NTZ; group 1) or prednisolone (groups 1 and 2) were assessed. Infection with T. foetus persisted in all cats for the entire 203-day study and resulted in diarrhea that resolved after 7 weeks. Group-1 cats had an earlier onset, more severe diarrhea, and increased number of trichomonads on direct fecal examination, compared with group-2 cats. Use of NTZ eliminated shedding of T. foetus and Cryptosporidium oocysts, but diarrhea consisting of trichomonad-containing feces recurred when treatment was discontinued. Prednisolone did not have an effect on infection with T. foetus but resulted in reappearance of Cryptosporidium oocysts in the feces of 2 of 4 cats. During necropsy, T. foetus was isolated from contents of the ileum, cecum, and colon. Tritrichomonas foetus organisms and antigen were detected on surface epithelia and within superficial detritus of the cecal and colonic mucosa. After experimental inoculation in cats, T. foetus organisms colonize the ileum, cecum, and colon, reside in close contact with the epithelium, and are associated with transient diarrhea that is exacerbated by coexisting cryptosporidiosis but not treatment with prednisolone.
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                Author and article information

                Contributors
                Journal
                Vet Parasitol
                Vet. Parasitol
                Veterinary Parasitology
                Elsevier B.V.
                0304-4017
                1873-2550
                29 July 2013
                8 November 2013
                29 July 2013
                : 197
                : 3
                : 418-426
                Affiliations
                [a ]Royal Canin Research Center, 650 Avenue de la Petite Camargue, 30470 Aimargues, France
                [b ]Université Paris-Est, Ecole Nationale Vétérinaire d’Alfort, 7 avenue du général de Gaulle, 94704 Maisons-Alfort Cedex, France
                [c ]Scanelis, 9 allée Charles Cros, 31771 Colomiers, France
                [d ]Institut Pasteur de Lille, Centre d’Infection et d’Immunité de Lille (CIIL), UMR CNRS 8204, Inserm U1019, Université Lille Nord deFrance, Biologie et Diversité des Pathogènes Eucaryotes Emergents, EA4547, 1 rue du Professeur Calmette, BP245, 59019 Lille cedex, France
                [e ]Microbe Division/Japan Collection of Microorganisms, RIKEN BioResource Center, 3-1-1 Koyadai, Tsukuba, Ibaraki 305-0074, Japan
                Author notes
                [* ]Corresponding author. Tel.: +33 466730632. aurelien.grellet@ 123456royalcanin.com
                Article
                S0304-4017(13)00435-4
                10.1016/j.vetpar.2013.07.030
                7116904
                23993636
                743d1afc-84b5-4de0-982b-f5f9b6c516d4
                Copyright © 2013 Elsevier B.V. All rights reserved.

                Since January 2020 Elsevier has created a COVID-19 resource centre with free information in English and Mandarin on the novel coronavirus COVID-19. The COVID-19 resource centre is hosted on Elsevier Connect, the company's public news and information website. Elsevier hereby grants permission to make all its COVID-19-related research that is available on the COVID-19 resource centre - including this research content - immediately available in PubMed Central and other publicly funded repositories, such as the WHO COVID database with rights for unrestricted research re-use and analyses in any form or by any means with acknowledgement of the original source. These permissions are granted for free by Elsevier for as long as the COVID-19 resource centre remains active.

                History
                : 21 February 2013
                : 17 July 2013
                : 20 July 2013
                Categories
                Article

                Parasitology
                trichomonads,pentatrichomonas hominis,dogs,france,epidemiology
                Parasitology
                trichomonads, pentatrichomonas hominis, dogs, france, epidemiology

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