Survey and Molecular Characterization of Trichomonads in Pigs in Anhui Province, East China, 2014.
ABSTRACT: Background:In pigs, several different trichomonad species such as Tritrichomonas foetus, Tetratrichomonas buttreyi, and Pentatrichomonas hominis have been described as inhabiting the digestive tract. However, little information is available on the epidemiology of these neglected parasites in the Chinese pig population. Methods:The prevalence of T. suis, T. buttreyi and P. hominis among 500 fecal specimens from pigs at seven pigs farms in Anhui Province in China between Oct and Dec 2014, was determined by PCR and DNA sequence analysis of the small subunit ribosomal RNA (SSU rRNA) genes. Results:The prevalence rates for T. suis, T. buttreyi, and P. hominis were 2.8% (14/500), 42.0% (210/500) and 7.8% (39/500), respectively. Mixed infections of two or three trichomonads were detected in 24 samples. The prevalence of the three trichomonads differed significantly between some age groups, with higher infection rates of T. suis and T. buttreyi in nursery pigs and P. hominis in preweaned pigs. The SSU rRNA sequences from T. suis and P. hominis showed 100% homology with their respective homologous database sequences. However, we observed minor allelic variations in the SSU rRNA sequences from T. buttreyi, and the five representative sequences identified were named firstly as types 1, 2, 3, 4 and 5. Moreover, type 1 was found to be dominant in the present study. Conclusion:These findings highlight the potential risk posed by pigs in the transmission of trichomonad infections to humans and other animals.
Project description:Trichomonad species inhabit a variety of vertebrate hosts; however, their potential zoonotic transmission has not been clearly addressed, especially with regard to human infection. Twenty-one strains of trichomonads isolated from humans (5 isolates), pigs (6 isolates), rodents (6 isolates), a water buffalo (1 isolate), a cow (1 isolate), a goat (1 isolate), and a dog (1 isolate) were collected in Indonesia and molecularly characterized. The DNA sequences of the partial 18S small subunit ribosomal RNA (rRNA) gene or 5.8S rRNA gene locus with its flanking regions (internal transcribed spacer region, ITS1 and ITS2) were identified in various trichomonads; Simplicimonas sp., Hexamastix mitis, and Hypotrichomonas sp. from rodents, and Tetratrichomonas sp. and Trichomonas sp. from pigs. All of these species were not detected in humans, whereas Pentatrichomonas hominis was identified in humans, pigs, the dog, the water buffalo, the cow, and the goat. Even when using the high-resolution gene locus of the ITS regions, all P. hominis strains were genetically identical; thus zoonotic transmission between humans and these closely related mammals may be occurring in the area investigated. The detection of Simplicimonas sp. in rodents (Rattus exulans) and P. hominis in water buffalo in this study revealed newly recognized host adaptations and suggested the existence of remaining unrevealed ranges of hosts in the trichomonad species.
Project description: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.
Project description:Three different parasites of the phylum Parabasala (Tritrichomonas foetus, Trichomitus rotunda and Tetratrichomonas buttreyi) have been described in pigs. In a previous study (Mostegl et al., 2011) approximately 47% of 91 paraffin wax-embedded intestinal samples of pigs which were Trichomonas-positive by in situ hybridization using a probe with a broad reactivity spectrum contained other species than T. foetus. Out of these, intestinal trichomonads from three pigs (pigs 1-3) were further analyzed by gene sequencing of a part of the 18S ribosomal RNA (rRNA) gene using primer walking. Subsequently, the partial sequences achieved by the different primer pairs were combined to a sequence of about 1000 bp for each trichomonad. In all three pigs unique sequences were acquired which showed only moderate similarities to sequences available in the GenBank. Alignments and the BLAST analysis showed a high degree of homology between sequences of trichomonads from pig 1 and pig 3 with only 1% difference. These sequences were found to be 92% similar to Hypotrichomonas acosta, a trichomonad isolated from squamate reptiles. The trichomonad sequence detected in the intestine of pig 2 showed about 10% nucleotide differences compared to pigs 1 and 3. This sequence was 97% similar to two Trichomitus batrachorum (a frog symbiont) sequences. A phylogenetic analysis using the neighbor-joining and maximum likelihood methods supported the data of the BLAST analysis. These results suggest the presence of at least two as yet undescribed trichomonad species in the intestinal contents of pigs.
Project description:Trichomonads are obligate anaerobes generally found in the digestive and genitourinary tract of domestic animals. In this study, four trichomonad isolates were obtained from carabao, dog, and pig hosts using rectal swab. Genomic DNA was extracted using Chelex method and the 18S rRNA gene was successfully amplified through novel sets of primers and undergone DNA sequencing. Aligned isolate sequences together with retrieved 18S rRNA gene sequences of known trichomonads were utilized to generate phylogenetic trees using maximum likelihood and neighbor-joining analyses. Two isolates from carabao were identified as Simplicimonas similis while each isolate from dog and pig was identified as Pentatrichomonas hominis and Trichomitus batrachorum, respectively. This is the first report of S. similis in carabao and the identification of T. batrachorum in pig using 18S rRNA gene sequence analysis. The generated phylogenetic tree yielded three distinct groups mostly with relatively moderate to high bootstrap support and in agreement with the most recent classification. Pathogenic potential of the trichomonads in these hosts still needs further investigation.
Project description:The trichomonad species Tritrichomonas foetus and Pentatrichomonas hominis were recently detected in the feces of dogs with diarrhea. However, little information is available on the prevalence and pathogenicity of these parasites in the canine population. Therefore, the aim of this study was to determine the prevalence and molecular characterization of trichomonads infecting pet dogs in Anhui and Zhejiang provinces, east China. In total, 315 pet dogs, with or without diarrhea, from 7 pet hospitals were included in this epidemiological survey. Microscopy and PCR detected P. hominis in 19.7% (62/315) and 31.4% (99/315) of fecal samples, respectively. T. foetus infection was detected in 0% (0/315) of samples with microscopy and in 0.6% (2/315) with PCR. The prevalence of P. hominis was significantly higher in young dogs (?12 months) than in adult dogs (>12 months), and was significantly higher in diarrheic dogs (50.6%) than in non-diarrheic dogs (24.3%; P<0.05). Infection with T. foetus did not correlate with any risk factors evaluated in this study. A sequence analysis of the P. hominis PCR products showed minor allelic variations between our sequences and those of P. hominis strains from other hosts in different parts of the world. Type CC1 was the most common strain in dogs in east China. The internal transcribed spacer 1 (ITS1)-5.8S rRNA gene sequences from the 2 T. foetus isolates detected in this study displayed 100% identity and were homologous to the sequences of other strains isolated from domestic cats in other countries.
Project description:Cryptosporidium spp., ubiquitous enteric parasitic protozoa of vertebrates, recently emerged as an important cause of economic loss and zoonosis. The present study aimed to determine the distribution and species of Cryptosporidium in post-weaned and adult pigs in Shaanxi province, northwestern China. A total of 1,337 fresh fecal samples of post-weaned and adult pigs were collected by sterile disposable gloves from 8 areas of Shaanxi province. The samples were examined by Sheather's sugar flotation technique and microscopy at × 400 magnification for Cryptosporidium infection, and the species in positive samples was further identified by PCR amplification of the small subunit (SSU) rRNA gene. A total of 44 fecal samples were successfully amplified by the nested PCR of the partial SSU rRNA, with overall prevalence of 3.3%. The average prevalence of Cryptosporidium infection in each pig farms ranged from 0 to 14.4%. Species identification by sequencing of SSU rRNA gene revealed that 42 (3.1%) samples were Cryptosporidium suis and 2 (0.15%) were Cryptosporidium scrofarum. C. suis had the highest prevalence (7.5%) in growers and the lowest in breeding pigs (0.97%). C. suis was the predominant species in pre-weaned and adult pigs, while C. scrofarum infected pigs older than 3 months only. A season-related difference of C. suis was observed in this study, with the highest prevalence in autumn (5.5%) and the lowest (1.7%) in winter. The present study provided basic information for control of Cryptosporidium infection in pigs and assessment of zoonotic transmission of pigs in Shaanxi province, China.
Project description:Trichomonadid protozoa have been found in the intestinal tracts of common marmosets (Callithrix jacchus). However, there is little information available on species identification and the pathogenicity of these trichomonads. In this study, we conducted a fecal survey of a common marmoset colony maintained as laboratory animals in Japan and identified the trichomonad species. Screening using a fecal smear examination revealed that 66% (58/88) of the marmosets had trichomonadid trophozoites in their feces. The trichomonads were found in both normal feces (31/49, 63%) and diarrhea (27/39, 69%), with no significant difference in frequency. The protozoa were identified as Pentatrichomonas hominis using morphological characters and the 100% identity of the nucleotide sequence of the partial 18S rRNA gene (297 bp). The intraspecific genetic variability between P. hominis from the marmosets in this study and P. hominis from other reported mammal hosts was ?1% in the nucleotide sequence, including the internal transcribed spacer (ITS)-1, 5.8S rRNA gene, and ITS-2 (293 bp). P. hominis inhabits the large intestine of various mammalian hosts, including primates, and is considered nonpathogenic. These results suggest that P. hominis is transmitted among marmosets and other mammals but is not a primary cause of bowel disease in marmosets.
Project description:In pigs, three different trichomonad species (Tritrichomonas foetus, Tetratrichomonas buttreyi and Tritrichomonas rotunda) have been described as commensals in the large intestine. The aim of this study was to gain further knowledge on the prevalence and pathogenicity of trichomonads in pigs by using a morphology-based approach. Chromogenic in situ hybridization (ISH) is a technique which allows direct localization of the protozoa in the intestinal tissue and correlation of the infection with pathologic changes. In the present study paraffin-wax embedded colon and ileum samples of 192 pigs were analyzed with this method. Using a probe specific for all known members of the order Trichomonadida (OT) 100 of the 192 pigs were tested positive. Thereof, about 10% showed moderate to high-grade parasitic load with trichomonads invading the lamina propria. Partial 18S ribosomal RNA gene sequencing of six of those animals showed a 100% sequence identity with T. foetus sequences. The majority of these animals were also tested positive for other enteropathogenic agents, such as Brachyspira sp., Lawsonia intracellularis, Escherichia coli, and porcine circovirus type 2. All OT-positive samples were further examined with another probe complementary to all known Tritrichomonas species sequences including T. foetus, T. augusta, T. mobilensis and T. nonconforma resulting in only 48 positives. These results suggest that T. foetus may not only be considered as an intestinal commensal but rather a facultative pathogen of pigs with a tendency for tissue invasion in the presence of other agents. Furthermore, the existence of other - yet to be identified - trichomonad species in the colon of pigs was shown.
Project description:BACKGROUND: Cryptosporidium spp. are common intestinal protozoa of humans and animals. There have been few studies conducted on the molecular characterizations of pig-derived Cryptosporidium isolates worldwide, especially in China. Thus, the aim of the present study was to understand the prevalence, distribution and genotypes of Cryptosporidium in pigs in Heilongjiang Province, China. METHODOLOGY/PRINCIPAL FINDINGS: A total of 568 fecal samples from pre-weaned and post-weaned piglets were collected from eight pig farms from four areas of Heilongjiang Province. The average infection rate of Cryptosporidium was 1.6% (9/568) by microscopy. 113 samples were subjected to PCR amplification of the small subunit (SSU) rRNA gene of Cryptosporidium, with 55.8% (63/113) being positive for Cryptosporidium. Cryptosporidium suis (n = 31) and C. scrofarumn (n = 32) were identified by DNA sequencing of the SSU rRNA gene. Three types of C. scrofarumn were found at the SSU rRNA locus, with one novel type being detected. Using species/genotype-specific primers for pig-adapted Cryptosporidium spp., 22 and 23 respectively belonged to C. suis and C. scrofarum mono-infections, with 18 co-infections detected. The infection peaks for C. suis (60%, 24/40) and C. scrofarum (51.2%, 21/41) were respectively found in the piglets of 5 to 8 weeks and more than 8 weeks. CONCLUSION/SIGNIFICANCE: The detection of C. suis and C. scrofarum in pre-weaned and post-weaned piglets has public health implications, due to the fact that the two species are both zoonotic Cryptosporidium. The novel C. scrofarum type detected may be endemic to China.
Project description:Trichomonads are anaerobic flagellated protists that, based on analyses of ribosomal RNA sequences, represent one of the earliest branching lineages among the eukaryotes. The absence of mitochondria in these organisms coupled with their deep phylogenetic position has prompted several authors to suggest that trichomonads, along with other deeply-branching amitochondriate protist groups, diverged from the main eukaryotic lineage prior to the endosymbiotic origin of mitochondria. In this report we describe the presence of a gene in Trichomonas vaginalis specifically related to mitochondrial chaperonin 60 (cpn60). A recent study indicates that a protein immunologically related to cpn60 is located in trichomonad hydrogenosomes. Together, these data provide evidence that ancestors of trichomonads perhaps harbored the endosymbiotic progenitors of mitochondria, but that these evolved into hydrogenosomes early in trichomonad evolution.