A freeze-fracture electron microscope study of Trichomonas vaginalis Donné and Tritrichomonas foetus (Riedmüller)

Two strains of Trichomonas vaginalis, JH162A , with low pathogenicity, and Balt 44, with high pathogenicity, as well as one highly pathogenic strain, KV-1, of Tritrichomonas foetus were studied by freeze-fracture electron microscopy. The protoplasmic faces ( PFs ) of the cell membranes of all three strains of both species had similar numbers of intramembranous particles (IMPs); however, the particles in the external faces (EFs) of these membranes were least abundant in Trichomonas vaginalis strain Balt 44 and most numerous in those of strain JH162A of this species. In Tritrichomonas foetus strain KV-1 the number of IMPs in the EF was close to but somewhat lower than that in the mild strain of the human urogenital trichomonad . In both species, the anterior, but not the recurrent, flagella had rosette-like formations, consisting of approximately 9 to 12 IMPs on both the PFs and EFs. The numbers and distribution of the rosettes appeared to vary among different flagella and in different areas of individual flagella of a single organism belonging to either species. The freeze-fracture electron micrographs provided a more complete understanding of the fine structure of undulating membranes of Trichomonadinae , as represented by Trichomonas vaginalis, and of Tritrichomonadinae (the Tritrichomonas augusta -type), as exemplified by Tritrichomonas foetus, than was gained from previous transmission and scanning electron microscope studies. Typically three longitudinal rows of IMPs on the PF of the recurrent flagellum of Trichomonas vaginalis were noted in the area of attachment of this flagellum to the undulating membrane. The functional aspects of the various structures and differences between certain organelles revealed in the two trichomonad species by the freeze-fracture method are discussed.     

Trichomonas vaginalis surface proteins: a view from the genome

Surface proteins of mucosal microbial pathogens play multiple and essential roles in initiating and sustaining the colonization of the heavily defended mucosa. The protist Trichomonas vaginalis is one of the most common human sexually transmitted pathogens that colonize the urogenital mucosa. However, little is known about its surface proteins. The recently completed draft genome sequence of T. vaginalis provides an invaluable resource to guide molecular and cellular characterization of surface proteins and to investigate their role in pathogenicity. Here, we review the existing data on T. vaginalis surface proteins and summarize some of the main findings from the recent in silico characterization of its candidate surface proteins.     

An ectonucleotide ATP-diphosphohydrolase activity in Trichomonas vaginalis stimulated by galactose and its possible role in virulence

This work describes the ability of living Trichomonas vaginalis to hydrolyze extracellular ATP (164.0 +/- 13.9 nmol Pi/h x 10(7) cells). This ecto-enzyme was stimulated by ZnCl2, CaCl2 and MgCl2, was insensitive to several ATPase and phosphatase inhibitors and was able to hydrolyze several nucleotides besides ATP. The activity was linear with cell density and with time for at least 60 min. The optimum pH for the T. vaginalis ecto-ATPase lies in the alkaline range. D-galactose, known to be involved in adhesion of T. vaginalis to host cells, stimulated this enzyme by more than 90%. A comparison between two strains of T. vaginalis showed that the ecto-ATPase activity of a fresh isolate was twice as much as that of a strain axenically maintained in culture, through daily passages, for several years. The results suggest a possible role for this ecto-ATPase in adhesion of T. vaginalis to host cells and in its pathogenicity.     

Growth of Trichomonas vaginalis in a serum-free McCoy cell culture system

Axenic cultures of Trichomonas vaginalis normally require serum for proliferation, yet serum-containing medium may interfere with the detection of T. vaginalis-secreted virulence factors. Trichomonas vaginalis can, however, grow in coculture with a McCoy cell monolayer in both the presence and absence of serum. For 6 T. vaginalis isolates examined, growth in this serum-free system shows lower peak concentrations of T. vaginalis and longer doubling times than those apparent in a serum-containing McCoy cell system. McCoy cells employed in the system did not appear to secrete soluble growth factors for T. vaginalis. The presence of McCoy cells was required for serum-free proliferation of T. vaginalis possibly indicating that eukaryotic cell membrane components may be important in supporting serum-free growth in this system.     

Non-physiological expression of UhpT does not lead to uncontrolled leakage of sugar phosphates out of Escherichia coli cells

Abstract A Trichomonas vaginalis cDNA library was constructed and recombinant plaques were screened using rabbit immunoglobulins specific for P65, a protozoan protein involved in pathogenicity that we identified in a previous study. A 1.38 kilobases cDNA fragment coding for the P65 protein was cloned in E. coli and then sequenced. On the basis of of the sequence obtained, six primers were synthesised and used to set up a Polymerase Chain Reaction. The presence of a specific amplicon in all 30 clinical isolates tested shows that P65 is a conserved and stable gene. The reaction is highly sensitive (as few as 5 to 10 parasites can be detected) and specific for Trichomonas vaginalis ; the gene coding P65 adhesin can be therefore considered a very good molecular target for polymerase chain reaction-based diagnostic purposes.     

中老年阴道炎患者阴道毛滴虫感染状况调查分析

目的:分析中老年阴道炎患者阴道毛滴虫感染情况,为中老年阴道炎的防治提供依据。方法:对2016年9月～2019年9月来我院就诊的中老年阴道炎患者的阴道分泌物进行阴道毛滴虫检查,同时通过问卷调查方法收集患者的病历资料,回收有效问卷200份,分析不同年龄、季节、职业、生活习惯、性生活情况患者的阴道毛滴虫感染状况。结果:中老年阴道炎患者阴道毛滴虫感染率为10.50%,其中45～50岁年龄段阴道炎患者阴道毛滴虫感染率最高,占19.15%,71～80岁年龄段阴道炎患者阴道毛滴虫感染率最低,占2.78%,不同年龄段阴道炎患者阴道毛滴虫感染率比较有统计学差异(P0.05)。春季、夏季阴道毛滴虫感染率显著高于秋季、冬季(P0.05)。牧民、农民阴道炎患者阴道毛滴虫感染率显著高于工人和公职人员的感染率(P0.05),牧民、农民阴道炎患者阴道毛滴虫感染率无明显差异(P0.05)。经常性清洁外阴的阴道炎患者阴道毛滴虫感染率显著低于非经常性清洁外阴的患者(P0.05)。有性生活阴道炎患者阴道毛滴虫感染率显著高于无性生活患者(P0.05),有性生活应用避孕套的阴道炎患者阴道毛滴虫感染率显著低于不应用避孕套患者(P0.05)。结论:中老年阴道炎患者阴道毛滴虫感染率较高,春季、夏季是阴道毛滴虫感染的高峰期,中年患者感染率高于老年患者,农民、牧民、服务人员和非经常性清洁外阴的患者阴道毛滴虫感染率较高,且有性生活没有应用避孕套者感染率更高,应做好以上高危人群的防治工作,以降低阴道毛滴虫的感染率。     

Flagellar duplication and migration during the Trichomonas vaginalis cell cycle

Trichomonas vaginalis is a flagellated protozoan, a representative of 1 of the earliest known eukaryotic lineages. Trichomonas vaginalis lacks centrioles but possesses basal bodies. We report here the cell cycle-dependent flagellar dynamics of T. vaginalis. By immunofluorescence, we found that T. vaginalis flagella duplicated during S-phase, segregated toward the nuclear poles, and then emanated from the spindle poles at mitosis. This behavior strongly parallels that of centrioles and other spindle pole-associated structures variously termed centrosomes, spindle pole bodies, or microtubule organizing centers. These observations support the hypothesis that flagellar forces may have provided motile forces for spindle pole alignment in an ancestral eukaryote.     

Association between Trichomonas vaginalis and some types of lesions of the uterine neck based on data collected in the quinquennium of 1974--1978]

In our statistic the number of cases of vaginal trichomoniasi in which discariosi, displasie, metaplasie are found are represented in about 40%. While 37,5% of women affected with neoplastic lesions had Trichomonas vaginalis. It is certain that in the presence of flogosi from Trichomonas vaginalis we meet more frequently than normal, atipie cellulars and that often an exact valutation of this lesion is put back to an exam after the recovery of the Trichomonas vaginalis, because fortunately, more than often these disturbances disappear.     

乳酸杆菌的代谢产物对体外培养的阴道毛滴虫的影响

目的研究乳酸杆菌代谢产物对体外培养的阴道毛滴虫的杀伤作用。方法检测不同浓度的乳酸杆菌代谢产物对体外培养的阴道毛滴虫在不同作用时间下的杀伤率。结果乳酸杆菌代谢产物浓度10%,作用时间2 h、4 h和6 h体外培养毛滴虫的杀伤率分别为26.43%、37.47%和46.35%;浓度25%时杀伤率分别为43.56%、74.65%和90.15%;浓度50%杀伤率分别为92.36%、95.23%和99.01%。结论乳酸杆菌代谢产物的浓度越高,对体外培养的毛滴虫的杀伤力越大,作用时间越长,效果越好。     

Analysis of surface saccharides in Trichomonas vaginalis strains with various pathogenicity levels by fluorescein-conjugated plant lectins

Certain surface saccharides of organisms from clone-derived cultures of five Trichomonas vaginalis strains, JH30A-cl. 1, JH31A-cl. 1, JH32A-cl. 1, JH34A-cl. 1, JH162A-cl. 1, and JH384A-cl. 2, which differed in their pathogenicity for women and experimental hosts, were compared with the aid of fluorescein-conjugated plant lectins using a quantitative fluorescence method. The lectins used were: concanavalin A (Con-A), wheat germ agglutinin (WGA), soybean agglutinin (SBA), castor bean agglutinin (CBA), and garden pea agglutinin (GPA). On the basis of experimental results and control experiments, the latter involving incorporation of specific inhibitory sugars in the reaction mixtures, it was concluded that: (1) All five strains had large numbers of Con-A- and WGA-binding saccharide residues. (2) Some also had smaller numbers of SBA- and CBA-binding sites. (3) No strain bound significant amounts of GPA. The differences in CBA binding were not related to pathogenicity of the parasites; however, those in SBA binding could be correlated with the pathogenicity levels of the five strains. The results obtained with SBA in the presence of N-acetyl-D-galactosamine and D-lactose and those recorded for GPA suggested that the differences between the pathogenic and mild T. vaginalis strains reflected the levels of D-lactosyl residues on the cell surfaces--these residues were more abundant on strains having higher pathogenicity levels. Possible explanation of the apparent relationships between the presence of the specific sugar residues and pathogenicity are suggested directly or by analogy with other pyranosyls (galactosyls).     

Characterization of Trichomonad Species and Strains by PCR Fingerprinting

ABSTRACT. The random amplified polymorphic DNA (RAPD) technique was used for phylogenetic analysis of trichomonads, for intraspecies genealogical study of Trichomonas vaginalis strains, and for assessment of intrastrain polymorphism in Trichomonas vaginalis . The phylogenetic tree for 12 trichomonad species showed certain discrepancies with current models of trichomonad evolution. However, it shows that RAPD traits retain phylogenetically relevant information. The results of intraspecies analyses of 18 Trichomonas vaginalis strains suggested some concordance between the genetic relationship of strains and their geographic origin. They also suggested a concordance between the strain genetic relationships and the resistance to metronidazole. A concordance was also found with respect to the severity of disease observed in donor patients but not with the results of laboratory virulence assays. No concordance was found between genetic relationship of strains and strain infection with a dsRNA Trichomonas vaginalis virus (TVV). The latter suggests that TVV might be transmitted horizontally among Trichomonas vaginalis populations. The identity of RAPD patterns of clones isolated from in vitro cultures and those of the cultures reisolated independently from the same patient within a period of six weeks suggests that individual Trichomonas vaginalis strains are not polymorphic and that the RAPD patterns are stable. Therefore, the RAPD technique seems useful for addressing various clinically relevant issues.     

蒙药金莲花体外抗阴道毛滴虫效果观察

目的观察金莲花对体外阴道毛滴虫的杀灭效果。方法采用不同浓度的金莲花水提物进行体外抗阴道毛滴虫试验,于药物作用后不同时间记录阴道毛滴虫的死亡率。结果金莲花具有抑制和杀灭阴道毛滴虫的作用,最低有效浓度为20.00 mg/mL。结论金莲花对阴道毛滴虫具有杀灭效果。     

探讨荆芥体外抗阴道毛滴虫的效果

目的探讨荆芥体外抗阴道毛滴虫的效果。方法将不同浓度的荆芥水提液作用于体外培养的阴道毛滴虫,于药物作用后不同时问记录毛滴虫的死亡率,并在光镜下观察药物作用前后毛滴虫的形态变化,同时与白头翁和青蒿的体外杀虫效果相比较。结果荆芥的杀虫效果与药物浓度和作用时间呈正相关。荆芥水提液触杀阴道毛滴虫的最低有效浓度为1：4。3种中药中,白头翁的杀虫效果最好,与另2种中药相比较差异有非常显著性（P〈0．01）;荆芥与青蒿的作用效果接近（P〉0．05）。荆芥作用后毛滴虫体内充满大量颗粒和空泡,部分虫体裂解、内容物外溢。结论荆芥具有较强的抗阴道毛滴虫作用。     

The influence of antimetabolites on Trichomonas vaginalis as an experimental model]

The present study was concerned with the relationship between antimetabolites and Trichomonas vaginalis. In all of the strains (with p-nitrobenzylether of N-acetyldiiodtryrosine) tested, the growth of Trichomonas vaginalis was identical to that of the control series at concentrations up to 2 mg/ml. At higher concentrations a decrease in multiplication was noted. In the experimental series with antitrypsin, from 1 mg/ml produced an inhibiting effect upon the Trichomonas population. Further investigations on the combined inhibition test are envisaged.     

Pathogenicity of Tritrichomonas mobilensis: subcutaneous inoculation in mice

The standard "subcutaneous mouse assay" was used to investigate the inherent pathogenicity of Tritrichomonas mobilensis, an intestinal parasite of squirrel monkeys. C57B1/6 mice given subcutaneous bilateral inocula of T. mobilensis died by day 4 postinoculation with lesions too small to be measured. Control mice similarly inoculated with pathogenic and nonpathogenic strains of Trichomonas vaginalis survived the challenge and produced lesions on day 6 with mean volumes in agreement with previous reports. CD1 mice similarly inoculated with standard and double doses of trichomonads (T. mobilensis) again produced small lesions. CD1 mice inoculated at double dosage were moribund or dead on days 5 and 6, respectively, postinoculation. Necropsies were performed on dead and sacrificed mice. Tissues were obtained from internal organs for histology and culture. Unexpectedly, trichomonads were cultured from liver and lung of C57B1/6 mice at the standard level of inoculation and liver, lung, and spleen of CD1 mice at the higher level of inoculation. Although trichomonads are normally considered surface-dwelling noninvasive organisms, the penetration of trichomonads to deep tissues is not without precedent. Tritrichomonas foetus and Trichomonas gallinae are known to invade tissues of their respective hosts. Trichomonas vaginalis has been demonstrated in subepithelial areas of both the prostate gland and cervix of humans. The ability of several species of trichomonads to invade tissues and/or migrate to other sites in their hosts suggests a need for revision of the concept of trichomonads as strictly lumen or surface-dwelling parasites.     

The effect of the growth medium composition on the fatty acids of Rhodothermus marinus and ''Thermus thermosphilus'' HB-8

Abstract We have investigated the mechanisms used by Trichomonas vaginalis to damage cellular membranes, using human erythrocytes as target cells. Haemolysis is a contact- and temperature-dependent phenomenon, and is inhibited in 4 mM EGTA. Osmotic protection experiments using carbohydrates with different molecular diameters as protectants demonstrated that the cytolytic activity of T. vaginalis is inhibited in 75 mM stachyose. On the basis of our data, we hypothesize a cytopathic mechanism mediated by the formation of functional pores into the target membrane. Some of the Trichomonas protein involved in haemolysis have been immunologically characterized.     

阴道毛滴虫铁氧还蛋白基因的原核表达

目的在原核细胞中表达阴道毛滴虫铁氧还蛋白(ferredoxin,Fd)基因。方法构建阴道毛滴虫Fd基因的原核表达重组质粒pUC19-Fd,转化大肠埃希菌JM109感受态细胞中,异丙基-β-D-硫代半乳糖苷(IPTG)诱导蛋白质表达。结果经十二烷基磺酸钠-聚丙烯酰胺凝胶电泳(SDS-PAGE)和免疫印迹(Western blot)分析,重组质粒在大肠埃希菌中表达出Fd。结论在大肠埃希菌中表达出了Fd。     

Interaction of Trichomonas vaginalis and Tritrichomonas foetus with keratin: an important role in parasite infection

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.     

荆芥体外抗阴道毛滴虫作用机制的实验研究

目的探讨荆芥体外杀灭阴道毛滴虫的作用机制。方法将浓度为1∶4的荆芥水提液作用于体外培养的阴道毛滴虫,并采用透射电镜观察经药物作用2 h和4 h后阴道毛滴虫的超微结构变化。结果荆芥作用后阴道毛滴虫多聚核糖体解聚,粗面内质网脱颗粒,胞质内可见大量空泡;随药物作用时间延长,核膜不完整,核质变疏松;最终,胞膜破损,内容物外溢,虫体死亡。结论荆芥可破坏阴道毛滴虫的内膜系统,具有较强的抗滴虫作用。