It is known that physicochemical conditions (e.g., pH, temperature, and ionic strength) affect the size of trichomonads. In this study, the sizes of 4 isolates of Trichomonas vaginalis cultured for more than a year (called "old T") and 3 isolates freshly isolated from vaginitis cases (called "fresh T") were compared by scanning electron microscopy. Although the fresh T had shorter body length, body width, and flagellar length than old T, total length (about 26 microm), including body length, flagella length, and axostyle length was almost the same in the 2 groups. A striking difference was observed between the axostyles of the 2 groups; the axostyle length of the fresh T (8.2 microm) was more than twice as long as that of the old T (4.0 microm). However, in several parasitology textbooks, the length of T. vaginalis is said to vary widely from 7 to 32 microm, and its undulating membrane is said to extend about half way (53.5%) to the posterior end of the body. On the other hand, in our study, the undulating membrane was observed to extend more than 3/4 of the body length (72.1%) in old T, whereas in fresh T it could not be measured. Taken together, we suggest that T. vaginalis averages 26 (21-32) microm in total length, with 9.5 (7.4-11.4) microm of body length and 6.8 (5.3-7.7) microm of width, and its undulating membrane extending 3/4 of its body length. Therefore, these findings may provide useful information for morphological characteristics of T. vaginalis.
*Biometry ; Female ; Humans ; *Microscopy, Electron, Scanning ; Organelles/ultrastructure ; Trichomonas Infections/parasitology ; Trichomonas vaginalis/*cytology/isolation & purification/*ultrastructure

Vaginal trichomoniasis, caused by Trichomonas vaginalis, is the most common sexually transmitted disease. More than 170 million people worldwide are annually infected by this protozoan. In the Republic of Korea, 10.4% of women complaining of vaginal symptoms and signs were found to be infected with T. vaginalis. However, despite its high prevalence, the pathogenesis of T. vaginalis infection has not been clearly characterized although neutrophil infiltration is considered to be primarily responsible for the cytologic changes associated with this infection. We hypothesized that trichomonads in the vagina sometime after an acute infection secrete proteins like excretorysecretory product that have a chemotactic effect on neutrophils, and that these neutrophils are further stimulated by T. vaginalis to produce chemokines like IL-8 and GRO-alpha, which further promote neutrophil recruitment and chemotaxis. Thus, neutrophil accumulation is believed to maintain or aggravate inflammation. However, enhanced neutrophil apoptosis induced by live T. vaginalis could contribute to resolution of inflammation. Macrophages may constitute an important component of host defense against T. vaginalis infection. For example, mouse macrophages alone and those activated by lymphokines or nitric oxide are known to be involved in the extracellular killing of T. vaginalis. In the host, T. vaginalis uses a capping phenomenon to cleave host immunoglobulins with proteinases and thus escape from host immune responses. Recently, we developed a highly sensitive and specific diagnostic polymerase chain reaction (PCR) technique using primers based on a repetitive sequence cloned from T. vaginalis (TV-E650), and found that the method enables the detection of T. vaginalis at concentrations as low as 1 cell per PCR mixture.
Trichomonas vaginalis/cytology/*isolation & purification/pathogenicity ; Trichomonas Vaginitis/drug therapy/*epidemiology/parasitology ; Korea/epidemiology ; Humans ; Female ; Animals

Neutrophils are the predominant inflammatory cells found in vaginal discharges of patients infected with Trichomonas vaginalis. In this study, we examined superoxide anion (O2(.-)) production by neutrophils activated by T. vaginalis. Human neutrophils produced superoxide anions when stimulated with either a lysate of T. vaginalis, its membrane component (MC), or excretory-secretory product (ESP). To assess the role of trichomonad protease in production of superoxide anions by neutrophils, T. vaginalis lysate, ESP, and MC were each pretreated with a protease inhibitor cocktail before incubation with neutrophils. Superoxide anion production was significantly decreased by this treatment. Trichomonad growth was inhibited by preincubation with supernatants of neutrophils incubated for 3 hr with T. vaginalis lysate. Furthermore, myeloperoxidase (MPO) production by neutrophils was stimulated by live trichomonads. These results indicate that the production of superoxide anions and MPO by neutrophils stimulated with T. vaginalis may be a part of defense mechanisms of neutrophils in trichomoniasis.
Anions/*metabolism ; Female ; Humans ; Neutrophils/enzymology/*metabolism/parasitology ; Peroxidase/metabolism ; Superoxides/*metabolism ; Trichomonas Infections/enzymology/*metabolism/parasitology ; Trichomonas vaginalis/*isolation & purification/physiology

Nongonococcal urethritis(NGU) is one of the common sexually transmitted diseases. Chlamydia trachomatis is the commonest pathogen of NGU. Ureaplasma urealyticum, Mycoplasma genitalium, Trichomonas vaginalis and other pathogens also account for some cases of NGU. With the development of molecular biology and immunology, more and more new techniques, such as PCR, LCR, etc., have been used in the researches on the laboratory diagnosis of NGU. It is necessary to establish and standardize some reliable rapid diagnostic tests for NGU. This paper reviews the progress in researches on the concept, etiology, clinical features, laboratory diagnosis and treatment of NGU.
Animals ; Clinical Laboratory Techniques ; Humans ; Mycoplasma Infections ; diagnosis ; drug therapy ; Trichomonas Infections ; diagnosis ; drug therapy ; Trichomonas vaginalis ; isolation & purification ; Ureaplasma Infections ; diagnosis ; drug therapy ; Ureaplasma urealyticum ; isolation & purification ; Urethritis ; diagnosis ; drug therapy

The objectives of this study were to conduct a prevalence survey of trichomoniasis in pregnant women and to evaluate the utility of different methods for its diagnosis. A total of 597 vaginal exudates from pregnant women who were examined at the Hospital de Clinicas in Buenos Aires, Argentina from 1 August 2005 to 31 January 2007, were prospectively and consecutively evaluated. The investigation of Trichomonas vaginalis was made by different microscopic examinations, and culture on liquid medium. The sensitivity and specificity of the microscopic examinations were assessed considering culture on liquid medium as the "gold standard". The prevalence of T. vaginalis obtained by culture on liquid medium was 4.0% (24/597). The prevalence of T. vaginalis obtained by direct wet smear, prolonged May-Grunwald Giemsa staining, and sodium acetate-formalin (SAF)/methylene blue staining-fixing technique was 1.8%, 2.3% and 2.5%, respectively. The sensitivity of the direct wet smear was 45.8%, that of the prolonged May-Grunwald Giemsa staining was 58.3%, and that of the SAF/methylene blue method was 62.5%. Considering the 3 microscopic examinations altogether, the sensitivity rose to 66.7% and the specificity was 100% for all of them. This is the first time that the prevalence data of T. vaginalis by culture in pregnant women are published in Argentina. Due to the low sensitivity obtained by microscopy in asymptomatic pregnant women, the use of the liquid medium is recommended during pregnancy, in order to provide an early diagnosis and treatment.
Argentina/epidemiology ; Cell Culture Techniques ; Female ; Humans ; Microscopy/*methods ; Parasitology/*methods ; Pregnancy ; Pregnancy Complications, Infectious/*diagnosis/*epidemiology/parasitology ; Pregnant Women ; Prevalence ; Prospective Studies ; Sensitivity and Specificity ; Trichomonas Infections/*diagnosis/*epidemiology/parasitology ; Trichomonas vaginalis/growth & development/*isolation & purification

The aim of this study was to assess the usefulness of PCR for diagnosis of Trichomonas vaginalis infection among male patients with chronic recurrent prostatitis and urethritis. Between June 2001 and December 2003, a total of 33 patients visited the Department of Urology, Hanyang University Guri Hospital and were examined for T. vaginalis infection by PCR and culture in TYM medium. For the PCR, we used primers based on a repetitive sequence cloned from T. vaginalis (TV-E650). Voided bladder urine (VB1 and VB3) was sampled from 33 men with symptoms of lower urinary tract infection (urethral charge, residual urine sensation, and frequency). Culture failed to detect any T. vaginalis infection whereas PCR identified 7 cases of trichomoniasis (21.2%). Five of the 7 cases had been diagnosed with prostatitis and 2 with urethritis. PCR for the 5 prostatitis cases yielded a positive 330 bp band from bothVB1 and VB3, whereas positive results were only obtained from VB1 for the 2 urethritis patients. We showed that the PCR method could detect T. vaginalis when there was only 1 T. vaginalis cell per PCR mixture. Our results strongly support the usefulness of PCR on urine samples for detecting T. vaginalis in chronic prostatitis and urethritis patients.
Adult ; DNA Primers/genetics ; Humans ; Male ; Middle Aged ; Molecular Diagnostic Techniques/*methods ; Parasitology/*methods ; Polymerase Chain Reaction/*methods ; Prostatitis/diagnosis/parasitology ; Republic of Korea ; Trichomonas Infections/*diagnosis/parasitology ; Trichomonas vaginalis/genetics/*isolation & purification ; Urethritis/diagnosis/parasitology

BACKGROUND: Next-generation sequencing (NGS) can detect many more microorganisms of a microbiome than traditional methods. This study aimed to analyze the vaginal microbiomes of Korean women by using NGS that included bacteria and other microorganisms. The NGS results were compared with the results of other assays, and NGS was evaluated for its feasibility for predicting vaginitis. METHODS: In total, 89 vaginal swab specimens were collected. Microscopic examinations of Gram staining and microbiological cultures were conducted on 67 specimens. NGS was performed with GS junior system on all of the vaginal specimens for the 16S rRNA, internal transcribed spacer (ITS), and Tvk genes to detect bacteria, fungi, and Trichomonas vaginalis. In addition, DNA probe assays of the Candida spp., Gardnerella vaginalis, and Trichomonas vaginalis were performed. Various predictors of diversity that were obtained from the NGS data were analyzed to predict vaginitis. RESULTS: ITS sequences were obtained in most of the specimens (56.2%). The compositions of the intermediate and vaginitis Nugent score groups were similar to each other but differed from the composition of the normal score group. The fraction of the Lactobacillus spp. showed the highest area under the curve value (0.8559) in ROC curve analysis. The NGS and DNA probe assay results showed good agreement (range, 86.2-89.7%). CONCLUSIONS: Fungi as well as bacteria should be considered for the investigation of vaginal microbiome. The intermediate and vaginitis Nugent score groups were indistinguishable in NGS. NGS is a promising diagnostic tool of the vaginal microbiome and vaginitis, although some problems need to be resolved.
Area Under Curve ; Bacteria/*genetics/isolation & purification ; Bacterial Proteins/genetics ; Candida/*genetics/isolation & purification ; Female ; Fungal Proteins/genetics ; Gardnerella vaginalis/genetics/isolation & purification ; High-Throughput Nucleotide Sequencing ; Humans ; *Microbiota ; RNA, Ribosomal, 16S/chemistry/genetics/metabolism ; ROC Curve ; Sequence Analysis, DNA ; Trichomonas vaginalis/genetics/isolation & purification ; Vagina/*microbiology ; Vaginitis/*diagnosis/microbiology

Trichomonas vaginalis, a causative agent of trichomoniasis, may trigger symptomatic or asymptomatic nongonococcal urethritis and chronic prostatitis in men. Despite the availability of highly sensitive diagnostic tests, such as nucleic acid amplification tests, including PCR, few prospective studies present data on male T. vaginalis infection in South Korea. In the present study, the prevalence of T. vaginalis and associated clinical conditions were evaluated in 201 male patients from a primary care urology clinic in South Korea. The prevalence of T. vaginalis infection in our cohort was 4% (8/201) by PCR. T. vaginalis infection was common in men older than 40 years (median age, 52 years). Among the 8 Trichomonas-positive patients, 87.5% (7/8) had prostatic diseases, such as prostatitis and benign prostatic hyperplasia, and 25.0% (2/8) and 12.5% (1/8) were coinfected with Chlamydia trachomatis and Mycoplasma genitalium, respectively. Our results suggest that T. vaginalis infection is not rare in men attending primary care urology clinics in South Korea, especially in those older than 40 years, in whom it may explain the presence of prostatic disease. The possibility of T. vaginalis infection should be routinely considered in older male patients with prostatic diseases in South Korea.
Adolescent ; Adult ; Aged ; Aged, 80 and over ; Child ; Chlamydia Infections/parasitology ; Chlamydia trachomatis/isolation & purification ; Coinfection ; Humans ; Male ; Middle Aged ; Mycoplasma Infections/parasitology ; Mycoplasma genitalium/isolation & purification ; *Polymerase Chain Reaction ; Prostatitis/epidemiology/*parasitology ; Republic of Korea/epidemiology ; Trichomonas Infections/*diagnosis ; Trichomonas vaginalis/*isolation & purification ; Young Adult

In this study, we describe Korean isolates of Trichomonas vaginalis infected with double-stranded (ds) RNA virus (TVV). One T. vaginalis isolate infected with TVV IH-2 evidenced weak pathogenicity in the mouse assay coupled with the persistent presence of a dsRNA, thereby indicating a hypovirulence effect of dsRNA in T. vaginalis. Cloning and sequence analysis results revealed that the genomic dsRNA of TVV IH-2 was 4,647 bp in length and evidenced a sequence identity of 80% with the previously-described TVV 1-1 and 1-5, but only a 42% identity with TVV 2-1 and 3 isolates. It harbored 2 overlapping open reading frames of the putative capsid protein and dsRNA-dependent RNA polymerase (RdRp). As previously observed in the TVV isolates 1-1 and 1-5, a conserved ribosomal slippage heptamer (CCUUUUU) and its surrounding sequence context within the consensus 14-nt overlap implied the gene expression of a capsid protein-RdRp fusion protein, occurring as the result of a potential ribosomal frameshift event. The phylogenetic analysis of RdRp showed that the Korean TVV IH-2 isolate formed a compact group with TVV 1-1 and 1-5 isolates, which was divergent from TVV 2-1, 3 and other viral isolates classified as members of the Giardiavirus genus.
Abscess/parasitology/pathology ; Animals ; Capsid Proteins/genetics ; Cloning, Molecular ; Disease Models, Animal ; Female ; Frameshifting, Ribosomal ; Giardiavirus/classification/genetics/*isolation & purification ; Humans ; Korea ; Mice ; Molecular Sequence Data ; Open Reading Frames ; Phylogeny ; RNA Replicase/genetics ; RNA, Double-Stranded/*genetics ; RNA, Viral/*genetics ; Sequence Analysis, DNA ; Sequence Homology ; Trichomonas Infections/*virology ; Trichomonas vaginalis/genetics/isolation & purification/pathogenicity/*virology ; Virulence

OBJECTIVETo investigate the value of computer-assisted slide-screening system (ThinPrep imaging system, TIS) in the diagnosis of cervical Thinprep smears.

METHODSA total of 19 600 ThinPrep smears were collected, including 9800 slides by TIS-assisted screening from September 2011 to March 2012 and 9800 slides by manual screening from September 2010 to April 2011 as control. The detection rates of abnormal cells and common microbial infection by the different screening methods were compared. With histopathological diagnosis of colposcopic biopsy as the gold standard, the screening efficiency and correlation of cytologic diagnosis among different screening methods were analyzed.

RESULTSCompared with manual screening, the detection rate of abnormal cells in 9800 cases by TIS-assisted screen was increased from 5.4% (525/9800) to 6.8% (665/9800), mainly in the categories of ASCUS and LSIL (P < 0.05). TIS had a higher accordance rate between cytologic diagnosis and histopathological diagnosis in the NILM and ASCUS than that by manual screening. False-negative rate of finding abnormal cells by TIS decreased from 8.5% (17/200) to 0.7% (2/289, P < 0.01) with an increased sensitivity compared to manual screening, although the specificity was similar. Both TIS and manual screening had advantages and disadvantages respectively in the detection of microbial organisms. TIS improved screening efficiency by 50%.

CONCLUSIONTIS improves not only the screening efficiency but also the detection of abnormal cells with a reduced false negativity, and it therefore has a broad application prospect.

Adenocarcinoma ; diagnosis ; pathology ; Adolescent ; Adult ; Aged ; Aged, 80 and over ; Candida ; isolation & purification ; Carcinoma, Squamous Cell ; diagnosis ; pathology ; Cervical Intraepithelial Neoplasia ; diagnosis ; pathology ; Cytodiagnosis ; False Negative Reactions ; Female ; Humans ; Image Processing, Computer-Assisted ; instrumentation ; Mass Screening ; Middle Aged ; Sensitivity and Specificity ; Trichomonas vaginalis ; isolation & purification ; Uterine Cervical Dysplasia ; diagnosis ; pathology ; Uterine Cervical Neoplasms ; diagnosis ; pathology ; Vaginal Smears ; Young Adult