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Microorganisms May 2024Multiple microbial detections in stool samples of indigenous individuals suffering from chronic gastroenteric disorder of a likely infectious origin, characterized by...
Collider Bias Assessment in Colombian Indigenous Wiwa and Kogui Populations with Chronic Gastroenteric Disorder of Likely Infectious Etiology Suggests Complex Microbial Interactions Rather Than Clear Assignments of Etiological Relevance.
Multiple microbial detections in stool samples of indigenous individuals suffering from chronic gastroenteric disorder of a likely infectious origin, characterized by recurring diarrhea of variable intensity, in the rural north-east of Colombia are common findings, making the assignment of etiological relevance to individual pathogens challenging. In a population of 773 indigenous people from either the tribe Wiwa or Kogui, collider bias analysis was conducted comprising 32 assessed microorganisms including 10 bacteria ( spp., spp., enteroaggregative (EAEC), enteropathogenic (EPEC), enterotoxigenic (ETEC), spp., Shiga toxin-producing (STEC), spp./enteroinvasive (EIEC), and spp.), 11 protozoa ( spp., spp., spp., , , /// complex, , , , and ), 8 helminths ( spp., , spp., , spp., spp., spp. and spp.), microsporidia ( spp.) and fungal elements (microscopically observed conidia and pseudoconidia). The main results indicated that negative associations potentially pointing towards collider bias were infrequent events (n = 14), while positive associations indicating increased likelihood of co-occurrence of microorganisms quantitatively dominated (n = 88). Microorganisms showing the most frequent negative associations were EPEC (n = 6) and spp. (n = 3), while positive associations were most common for spp. (n = 16), (n = 15), spp./EIEC (n = 12), spp. (n = 11) and spp. (n = 10). Of note, positive associations quantitively dominated for spp. In conclusion, collider bias assessment did not allow clear-cut assignment of etiological relevance for detected enteric microorganisms within the assessed Colombian indigenous population. Instead, the results suggested complex microbial interactions with potential summative effects. Future studies applying alternative biostatistical approaches should be considered to further delineate respective interactions.
PubMed: 38792799
DOI: 10.3390/microorganisms12050970 -
Journal of Parasitology Research 2013Trichomonads are obligate anaerobes generally found in the digestive and genitourinary tract of domestic animals. In this study, four trichomonad isolates were obtained...
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.
PubMed: 23936631
DOI: 10.1155/2013/831947 -
Journal of Clinical Microbiology Apr 1991Trichomoniasis is one of the most widespread sexually transmitted diseases in the world. Diagnosis can be achieved by several methods, such as direct microscopic...
Trichomoniasis is one of the most widespread sexually transmitted diseases in the world. Diagnosis can be achieved by several methods, such as direct microscopic observation of vaginal discharge, cell culture, and immunological techniques. A 2.3-kb Trichomonas vaginalis DNA fragment present in strains from diverse geographic areas was cloned and used as a probe to detect T. vaginalis DNA in vaginal discharge by a dot blot hybridization technique. This probe was specific for T. vaginalis DNA. It recognized strains from two regions in Italy (Sardinia, Piemonte) and from Mozambique (Africa). In addition, our probe did not cross-react with bacterial (Escherichia coli, Enterococcus spp., group B streptococci, Gardnerella vaginalis, Neisseria gonorrhoeae, Chlamydia trachomatis, and Lactobacillus spp.), viral (herpes simplex virus type 2), fungal (Candida albicans), protozoan (Entamoeba histolytica, Giardia lamblia, Plasmodium falciparum, Leishmania major, and Leishmania infantum), or human nucleic acids. The probe reacted with Pentatrichomonas hominis and Trichomonas foetus. The limit signal recognized by our probe corresponded to the DNA of 200 T. vaginalis isolates. The 2.3-kb probe was used in a clinical analysis of 98 samples. Of these, 20 samples were found to be positive both with the probe and by cell culture, and only 14 of these were positive by a standard wet mount method.
Topics: Animals; Cloning, Molecular; DNA Probes; DNA, Protozoan; Female; Humans; Trichomonas Vaginitis; Trichomonas vaginalis; Vaginal Smears
PubMed: 1890171
DOI: 10.1128/jcm.29.4.702-706.1991 -
Journal of the American Veterinary... May 2003To evaluate the efficacy of and optimize a commercially available culture system for sensitive and specific in-clinic culture of Tritrichomonas foetus from cat feces.
OBJECTIVE
To evaluate the efficacy of and optimize a commercially available culture system for sensitive and specific in-clinic culture of Tritrichomonas foetus from cat feces.
DESIGN
Prospective study.
SAMPLE POPULATION
Samples of freshly voided feces from 117 purebred cats and pure cultures of T. foetus obtained from a cat with chronic diarrhea.
PROCEDURE
Optimal conditions for use of the culture system, such as quantity of fecal inoculum (0.025 to 0.2 g) and cultivation temperature (25 or 37 degrees C [98.6 or 77.0 degrees F]), were determined. Specificity of the system was examined by attempted culture of Giardia lamblia and Pentatrichomonas hominis. Sensitivity of the system to detect T. foetus was determined by inoculation of culture system pouches with serially diluted T. foetus suspensions with and without feces.
RESULTS
Detection limit of the culture system was 1 and 1,000 T. foetus organisms without and with feces from cats, respectively. Optimal fecal inoculum was < 0.1 g of feces. At 37 degrees C, cultures yielded positive results in 24 hours; organisms remained viable for 1 to 6 days, and bacterial overgrowth was common. At 25 degrees C, cultures yielded positive results in 1 to 11 days; organisms were long-lived, and bacterial overgrowth was uncommon. Neither G. lamblia or P. hominis survived in the culture system.
CONCLUSIONS AND CLINICAL RELEVANCE
The culture system was sensitive and specific for culture of T. foetus in feces of cats. Performance was optimal when test kits were inoculated with < or = 0.1 g of freshly voided feces and cultured at 25 degrees C.
Topics: Animals; Cat Diseases; Cats; Feces; Female; Prospective Studies; Protozoan Infections; Protozoan Infections, Animal; Sensitivity and Specificity; Temperature; Time Factors; Tritrichomonas foetus
PubMed: 12762381
DOI: 10.2460/javma.2003.222.1376 -
PloS One 2018Intestinal parasitic infections are considered a serious public health problem and widely distributed worldwide, mainly in urban and rural environments of tropical and...
BACKGROUND
Intestinal parasitic infections are considered a serious public health problem and widely distributed worldwide, mainly in urban and rural environments of tropical and subtropical countries. Globally, soil-transmitted helminths and protozoa are the most common intestinal parasites. Blastocystis sp. is a highly prevalent suspected pathogenic protozoan, and considered an unusual protist due to its significant genetic diversity and host plasticity.
METHODOLOGY/MAIN FINDINGS
A total of 294 stool samples were collected from inhabitants of three rural valleys in Rio de Janeiro, Brazil. The stool samples were evaluated by parasitological methods, fecal culture, nested PCR and PCR/Sequencing. Overall prevalence by parasitological analyses was 64.3% (189 out of 294 cases). Blastocystis sp. (55.8%) was the most prevalent, followed by Endolimax nana (18.7%), Entamoeba histolytica complex (7.1%), hookworm infection (7.1%), Entomoeba coli (5.8%), Giardia intestinalis (4.1%), Iodamoeba butchilii (1.0%), Trichuris trichiura (1.0%), Pentatrichomonas hominis (0.7%), Enterobius vermicularis (0.7%), Ascaris lumbricoides (0.7%) and Strongyloides stercoralis (0.7%). Prevalence of IPIs was significantly different by gender. Phylogenetic analysis of Blastocystis sp. and BLAST search revealed five different subtypes: ST3 (34.0%), ST1 (27.0%), ST2 (27.0%), ST4 (3.5%), ST8 (7.0%) and a non-identified subtype.
CONCLUSIONS/SIGNIFICANCE
Our findings demonstrate that intestinal parasite infection rates in rural areas of the Sumidouro municipality of Rio de Janeiro, Brazil are still high and remain a challenge to public health. Moreover, our data reveals significant genetic heterogeneity of Blastocystis sp. subtypes and a possible novel subtype, whose confirmation will require additional data. Our study contributes to the understanding of potential routes of transmission, epidemiology, and genetic diversity of Blastocystis sp. in rural areas both at a regional and global scale.
Topics: Adolescent; Adult; Aged; Aged, 80 and over; Animals; Base Sequence; Blastocystis; Blastocystis Infections; Brazil; Child; Child, Preschool; Cross-Sectional Studies; DNA, Protozoan; Feces; Female; Genetic Variation; Helminthiasis; Humans; Intestinal Diseases, Parasitic; Male; Middle Aged; Phylogeny; Polymerase Chain Reaction; Prevalence; Protozoan Infections; Ribotyping; Rural Population; Sequence Alignment; Sequence Analysis, DNA; Sequence Homology, Nucleic Acid; Young Adult
PubMed: 29522552
DOI: 10.1371/journal.pone.0193860 -
Journal of Clinical Microbiology Aug 1988Tritrichomonas mobilensis is a recently described enteric protozoon of squirrel monkeys. An earlier report identified one of the metabolic products of this organism as a...
Tritrichomonas mobilensis is a recently described enteric protozoon of squirrel monkeys. An earlier report identified one of the metabolic products of this organism as a lectinlike hemagglutinin. Its further properties were determined in this study. Culture supernatants of T. mobilensis FP4190 were concentrated by ultrafiltration through a membrane with 100,000-molecular-weight cutoff. High titers of agglutinin against human erythrocytes were obtained. Incubation at 70 degrees C for 15 min resulted in complete inactivation. Exposure to 56 degrees C for 30 min was without effect, and only partial loss of activity was obtained during incubation for up to 18 h. Maintenance at pH 4 to 9 for 4 h at room temperature had no deleterious effect. Apparent degradation of the hemagglutinin was achieved by 18 h of contact with proteinase K, but trypsin and collagenase were essentially ineffective. Papain increased the sensitivity of the test. In the presence of this enzyme hemagglutinin was demonstrated also in cultures of Tritrichomonas foetus and Tritrichomonas augusta but not in those of Pentatrichomonas hominis or Trichomonas vaginalis.
Topics: Animals; Hemagglutination Tests; Hemagglutinins; Hot Temperature; Humans; Hydrogen-Ion Concentration; Papain; Species Specificity; Tritrichomonas
PubMed: 3170709
DOI: 10.1128/jcm.26.8.1460-1463.1988 -
Annals of Parasitology 2016A loop-mediated isothermal amplification (LAMP) assay was developed to detect the sexually-transmitted parasite, Trichomonas vaginalis in vaginal swabs. The presence of...
A loop-mediated isothermal amplification (LAMP) assay was developed to detect the sexually-transmitted parasite, Trichomonas vaginalis in vaginal swabs. The presence of T. vaginalis was detected from 121 female sex workers attending a social hygiene clinic in Balibago, Angeles City, Pampanga, Philippines using culture, polymerase chain reaction (PCR), and the developed LAMP assay. The high analytical sensitivity of LAMP detected a higher prevalence of T. vaginalis (42.06%) compared to culture (8.26%) and PCR (7.44%). Additionally, this assay did not cross-react with DNAs of other trichomonads that can infect humans such as Trichomonas tenax and Pentatrichomonas hominis as well as the pathogens, Candida albicans and Staphylococcus aureus. The LAMP assay developed had a limit of detection (0.036 ng/μl) lower than that of PCR using the primers TvK3 and TvK7 (0.36 ng/μl). Prevalence of T. vaginalis in female sex workers in this area of the Philippines may be higher than previously estimated. Discordant results of PCR and LAMP may be due to different reactions to different kinds of inhibitors in the vaginal swabs.
Topics: DNA, Protozoan; Female; Humans; Nucleic Acid Amplification Techniques; Philippines; Sex Workers; Time Factors; Trichomonas Vaginitis; Trichomonas vaginalis
PubMed: 27262954
DOI: 10.17420/ap6201.28 -
Veterinary Parasitology Jun 2012Trichomonads have been infrequently reported in the feces of dogs where their pathogenicity remains uncertain. It is currently unknown whether Tritrichomonas foetus or...
Trichomonads have been infrequently reported in the feces of dogs where their pathogenicity remains uncertain. It is currently unknown whether Tritrichomonas foetus or Pentatrichomonas hominis is identified more commonly in dogs with trichomonosis or how often these infections are accompanied by concurrent enteric infectious agents. The objective of this study was to determine the identity of trichomonads present in a series of 38 unsolicited canine diarrheic fecal samples submitted for T. foetus diagnostic polymerase chain reaction (PCR) testing between 2007 and 2010. We also examined each fecal sample for an association of trichomonosis with concurrent infection using a convenient real-time PCR panel for nine gastrointestinal pathogens. P. hominis, T. foetus, or both were identified by PCR in feces of 17, 1, and 1 dogs respectively. Feces from the remaining 19 dogs were PCR negative for T. foetus, P. hominis and using broader-spectrum Trichomonadida primers. The total number and specific identities of concurrent enteropathogens identified did not differ between fecal samples from dogs that were or were not identified by PCR as infected with trichomonads. These results suggest that P. hominis infection is more frequently identified than T. foetus infection in diarrheic dogs with trichomonosis and that concurrent enteropathogen infection is common in this population.
Topics: Animals; Coinfection; Diarrhea; Dog Diseases; Dogs; Feces; Female; Male; Trichomonas Infections
PubMed: 22264747
DOI: 10.1016/j.vetpar.2011.12.031 -
Journal of Clinical Microbiology Jan 1997A colorimetric one-tube nested PCR was developed for the detection of Trichomonas vaginalis in clinical vaginal discharge specimens. A family of 650-bp specific DNA...
A colorimetric one-tube nested PCR was developed for the detection of Trichomonas vaginalis in clinical vaginal discharge specimens. A family of 650-bp specific DNA repeats from the T. vaginalis genome was targeted. There was no cross-reaction with human DNA or other infectious agents, including Pentatrichomonas hominis and Giardia lamblia. The colorimetric assay was applied as an adjunct to nested PCR for semiquantitative determination of T. vaginalis DNA at levels corresponding to 1 to 1,000 parasites. PCR of samples prepared by a rapid boiling method was as sensitive and specific as PCR of samples prepared by the standard DNA extraction method: the equivalent of one T. vaginalis organism in 20 microliters of vaginal discharge could be detected. The colorimetric nested PCR was compared with wet mount and culture for the detection of T. vaginalis. A total of 378 clinical vaginal discharge specimens from symptomatic patients were examined; 31 patients were positive for T. vaginalis both by culture and by nested PCR. However, only 17 of these 31 patients were positive by wet mount examination. In addition, of 113 asymptomatic patients, 9 were positive for T. vaginalis by nested PCR. Of these nine PCR-positive patients, only two were also positive both by wet mount and by culture, four patients were positive by culture but negative by wet mount, and three patients were negative both by wet mount and by culture. No specimens negative by nested PCR were positive by wet mount or by culture. The three asymptomatic patients with PCR-positive but wet mount- and culture-negative samples were subsequently found to have T. vaginalis infection after repeated and prolonged culture was performed. This colorimetric nested PCR was very sensitive compared with culture for the diagnosis of vaginal trichomoniasis, especially asymptomatic T. vaginalis infection. It is also simple, specific, rapid, and semiquantitative.
Topics: Animals; Female; Humans; Polymerase Chain Reaction; Trichomonas Infections; Trichomonas vaginalis; Vagina; Vaginal Discharge
PubMed: 8968894
DOI: 10.1128/jcm.35.1.132-138.1997 -
Cellular Microbiology Jun 2005Trichomonas vaginalis, an ancient protist, colonizes the vaginal mucosa causing trichomonosis, a vaginitis that sometimes leads to severe health complications....
Trichomonas vaginalis, an ancient protist, colonizes the vaginal mucosa causing trichomonosis, a vaginitis that sometimes leads to severe health complications. Preparatory to colonization of the vagina is the adhesion to vaginal epithelial cells (VECs) by trichomonads. We hypothesized that VECs alter the gene expression to form a complex signalling cascade in response to trichomonal adherence. In order to identify the genes that are upregulated, we constructed a subtraction cDNA library after contact with parasites that is enriched for differentially expressed genes from the immortalized MS-74 VECs. Sixty cDNA clones were sequenced and to our knowledge for the first time, differentially regulated genes were identified in response to early trichomonal infection. The identified genes were found to encode functional proteins with specific functions associated with cell structure maintenance and extracellular matrix components, proinflammatory molecules and apoptosis. Semi-quantitative reverse transcription polymerase chain reaction (RT-PCR) confirmed expression of selected genes. Further, cyclooxygenase 2 (COX-2) protein expression was analysed using Western blot and immunofluorescence assays. Data suggest that p38 mitogen-activated protein (MAP) kinase and tyrosine kinases play a role in COX-2 induction. Finally, T. vaginalis and Tritrichomonas foetus but not Pentatrichomonas hominis induce expression of COX-2. This is a first attempt at elucidating the basis of interaction of trichomonads with host cells and the corresponding host responses triggered by the parasites.
Topics: Animals; Cell Adhesion; Cells, Cultured; Cyclooxygenase 2; Epithelial Cells; Female; Gene Expression Regulation; Humans; Membrane Proteins; Phosphotransferases; Prostaglandin-Endoperoxide Synthases; Reverse Transcriptase Polymerase Chain Reaction; Trichomonas vaginalis; Tritrichomonas foetus; Vagina
PubMed: 15888089
DOI: 10.1111/j.1462-5822.2005.00522.x