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electrophoretic analysis of soluble proteins and esterase, superoxide dismutase and acid phosphatase isoenzymes of members of the protozoan family trichomonadidae.polyacrylamide gel electrophoresis was used to compare the proteins and isoenzymes of esterase, superoxide dismutase, and acid phosphatase in soluble, whole-cell extracts of four strains of trichomonas vaginalis, two strains of trichomonas gallinae, and one strain each of tritrichomonas foetus, tritrichomonas augusta, tetratrichomonas gallinarum, and pentatrichomonas hominis. intraspecific, interspecific, and intergeneric differences were found in protein and isoenzyme profiles. at least four to ...19854017542
tetratrichomonas gallinarum in the blood of chickens infected with spirochetes. 19665953851
tetratrichomonas gallinarum encephalitis in a mockingbird (mimus polyglottos). 19969026073
molecular phylogeny of parabasalids based on small subunit rrna sequences, with emphasis on the trichomonadinae subfamily.we determined small subunit ribosomal dna sequences from three parabasalid species, trichomitus batrachorum strain r105, tetratrichomonas gallinarum, and pentatrichomonas hominis belonging to the trichomonadinae subfamily. unrooted molecular phylogenetic trees inferred by distance, parsimony, and likelihood methods reveal four discrete clades among the parabasalids. the trichomonadinae form a robust monophyletic group. within this subfamily t. gallinarum is closely related to trichomonas species ...200010651299
axenic culture of tetratrichomonas gallinarum: growth and anaerobic utilization of carbohydrates and related compounds. 196414213220
development of a blocking-elisa for the detection of antibodies against histomonas meleagridis in chickens and turkeys.histomonosis is an infectious disease of mainly galliform birds that can cause high mortality, especially in commercial turkey flocks. several diagnostic tools were available to detect its causative agent, the flagellated protozoan histomonas meleagridis. however, serological tools were largely lacking. recently, an indirect elisa has been described, but this test was not validated for specificity and might suffer from cross-reactivity with related protozoa. therefore, a specific blocking-elisa ...201020400229
design and validation of an oligonucleotide probe for the detection of protozoa from the order trichomonadida using chromogenic in situ hybridization.infections with protozoal parasites of the order trichomonadida are often observed in veterinary medicine. based on the trichomonad species involved these infections are either asymptomatic or can lead to sometimes serious disease. to further study protozoal agents of the order trichomonadida the establishment of a method to detect trichomonads directly in the tissue, allowing parasite-lesion correlation, is necessary. here we describe the design and evaluation of an oligonucleotide probe for ch ...201020395049
effect of peganum harmala or its beta-carboline alkaloids on certain antibiotic resistant strains of bacteria and protozoa from poultry.in the present study the antimicrobial potential of various extracts from 12 medicinal plants has been investigated in vitro on multiple antibiotic resistant pathogens and some selected protozoa isolated from poultry. the initial examination was performed on e. coli (n = 10) using disc and agar well diffusion assays. only two plants, peganum harmala (seeds) and eucalyptus globulus (leaves) showed positive responses. the active extracts were also investigated against an additional 19 bacteria and ...200818814210
first report of typhlitis/typhlohepatitis caused by tetratrichomonas gallinarum in three duck species.two red-breasted mergansers (mergus serrator), one hooded merganser (lophodytes cucullatus), and one common eider (somateria mollissima) from a german zoological collection died of necrotizing typhlitis/typhlohepatitis within 2 years. using a newly established chromogenic in situ hybridization assay, numerous intralesional trophozoites of tetratrichomonas gallinarum could be detected in formalin-fixed and paraffin-embedded tissues from the caeca and livers of the affected birds. partial sequenci ...201021154060
in-situ hybridization for the detection and identification of histomonas meleagridis in tissues.for use in an in-situ hybridization method, three probes (hm, tr and bl) were designed to hybridize, respectively, with (1) histomonas meleagridis, (2) tetratrichomonas gallinarum, and (3) a broad range of micro-organisms, including blastocystis spp. mono-eukaryotic cultures were used to test the specificity of the three oligonucleotides and to optimize the hybridization procedure before applying the probes to archived samples of various tissues and to a culture of trichomonas gallinae. specific ...200617069832
phylogenetic relationships of class ii fumarase genes from trichomonad species.class ii fumarase sequences were obtained by polymerase chain reaction from five trichomonad species. all residues known to be highly conserved in this enzyme were present. nuclear run-on assays showed that one of the two genes identified in tritrichomonas foetus was expressed, whereas no fumarase transcripts were detected in the related species trichomonas vaginalis. these findings corroborate previous biochemical data. fumarase genes were also expressed in monocercomonas sp. and tetratrichomon ...200111470849
comparative analysis of trichomonad genome sizes and karyotypes.in parasitic protists, the genome sizes range from 2.9mb in encephalitozoon cuniculi to about 160mb in trichomonas vaginalis. the suprisingly large genome size of the former human parasite resulted from the expansion of various repetitive elements, specific gene families, and possibly from large-scale genome duplication. the reason for this phenomenon, as well as whether other trichomonad species have undergone a similar genome expansion, is not known. in this work we studied the genomes of nine ...200818606195
pcr for the identification and differentiation of histomonas meleagridis, tetratrichomonas gallinarum and blastocystis spp.in the present investigation pcr assays were developed for the rapid detection and differentiation of two poultry flagellates: histomonas meleagridis and tetratrichomonas gallinarum as well as the protozoan microorganism: blastocystis spp. the nucleotide sequences of the small subunit ribosomal rnas were used for primer construction obtaining fragments which vary in size for each microorganism. the established pcrs were able to detect dna obtained from one microorganism of t. gallinarum and blas ...200616920265
clonal cultures of histomonas meleagridis, tetratrichomonas gallinarum and a blastocystis sp. established through micromanipulation.clonal cultures of histomonas meleagridis, tetratrichomonas gallinarum and a blastocystis sp. were established for the first time. single microbes were successfully isolated from a mixture of micro-organisms obtained from caecal contents of turkeys, using a micromanipulation approach. the cloned parasites were propagated in vitro and maintained through continuous passages multiplying to high numbers. identification of the protists was done by morphological investigation identifying various forms ...200616854251
tetratrichomonads from the oral cavity and respiratory tract of humans.to clarify the taxonomy of trichomonads associated with human respiratory diseases, we examined a collection of axenic trichomonad strains isolated from the oral cavity and bronchi of patients from pulmonary diseases clinics in tallin, estonia. the oral and bronchial strains were compared mutually as well as with a reference strain of trichomonas tenax, a common inhabitant of the human oral cavity, and other trichomonad species from humans and animals. unexpectedly, the morphological studies, as ...200516178352
phylogenetic relationships of the glycolytic enzyme, glyceraldehyde-3-phosphate dehydrogenase, from parabasalid flagellates.over 90% of the open reading frame of gap genes for glycolytic glyceraldehyde-3-phosphate dehydrogenase (gapdh; ec 1.2.1.12) was obtained with pcr from five species of parabasala. with gap1 from trichomonas vaginalis obtained earlier, the data include two sequences each for three species. all sequences were colinear with t. vaginalis gap1 and shared with it as a synapomorphy a 10- to 11-residue insertion not found in any other gap and an s-loop with characteristic features of eubacterial gapdh. ...19989694668
fine structure of the bird parasites trichomonas gallinae and tetratrichomonas gallinarum from cultures.the trophozoites of trichomonas gallinae and tetratrichomonas gallinarum were studied by means of light and electron microscopy after cloning and cultivating them axenically. t. gallinae trophozoites varied in shape reaching from ovoidal to pyriform and had a size of about 7-11 microm. they were provided with four free flagella and a fifth recurrent one, which did not become free at the posterior pole. the nucleus was ovoid, had a size of about 2.5-3 microm, and was situated closely below the ba ...200919421777
antiprotozoal activities determined in vitro and in vivo of certain plant extracts against histomonas meleagridis, tetratrichomonas gallinarum and blastocystis sp.a total of 43 plant substances provided as raw material and different kinds of extracts (aqueous, ethanol, and heptane) from 18 different organic wastes obtained from the food/feed industry were investigated for their in vitro activities against clonal cultures of histomonas meleagridis, tetratrichomonas gallinarum, and blastocystis sp. ethanolic extracts of thyme, saw palmetto, grape seed, and pumpkin fruit proved to be most efficacious. thus, these extracts were further tested in vivo in turke ...200818751730
histomonas meleagridis: immunohistochemical localization of parasitic cells in formalin-fixed, paraffin-embedded tissue sections of experimentally infected turkeys demonstrates the wide spread of the parasite in its host.in the present investigation, a polyclonal antibody-based immunohistochemical technique was developed to localize histomonas meleagridis in formalin-fixed, paraffin-embedded tissues of experimentally infected turkeys. the developed technique was highly specific for histomonads as no immunohistochemical reaction was observed with cultures of tetratrichomonas gallinarum, trichomonas gallinae and blastocystis sp. in addition, tissues positive for various other protozoan parasites and fungi were als ...200818155698
differences in the in vitro susceptibility of mono-eukaryotic cultures of histomonas meleagridis, tetratrichomonas gallinarum and blastocystis sp. to natural organic compounds.currently, all pharmaceuticals for the treatment or prophylaxis of blackhead disease (histomonosis) caused by the flagellate histomonas meleagridis are banned from the market. consequently, great interest exists on the finding of alternative drugs for the abatement of histomonosis. in this study, carvacrol, cassia oil, an essential oil (eo) mixture containing thyme and rosemary eo and a quillaja saponaria saponin were examined using in vitro assays for antiprotozoal and antibacterial activity te ...200717285346
detection of histomonas meleagridis in turkeys cecal droppings by pcr amplification of the small subunit ribosomal dna sequence.histomonas meleagridis is a protozoan parasite that may cause histomoniasis, a disease of gallinaceous fowl characterized by necrotic typhlitis, hepatitis and high mortality. diagnosis of this disease is based on direct identification or on cultivation of the parasite. with the aim of developing more sensitive, rapid and useful tools for parasite detection, pcr that amplified a dna target of 209 pb of the 18s rrna gene was designed to detect the genome of h. meleagridis and to differentiate it f ...200515979800
molecular phylogeny of trichomonadidae family inferred from its-1, 5.8s rrna and its-2 sequences.the trichomonads have been the subject of several molecular studies that reported some discrepancies both at the lower and higher taxonomic levels. the purpose of this study was to make an extensive phylogenetic analysis of the trichomonadidae using its-1/5.8s/its-2 sequences, to better understand its phylogeny and the usefulness of this marker. its-1/5.8s/its-2 sequences of 36 strains from 14 species belonging to trichomonadidae and monocercomonadidae were analysed, in which 20 were newly deter ...200415217735
in vitro effect of essential oils from cinnamomum aromaticum, citrus limon and allium sativum on two intestinal flagellates of poultry, tetratrichomonas gallinarum and histomonas meleagridis.essential oils may be effective preventive or curative treatments against several flagelated poultry parasites and may become primordial either to organic farms, or as more drugs are bannished. the anti-flagellate activity of essential oils obtained from fresh leaves of cinnamomum aromaticum, citrus limon pericarps and allium sativum bulbs was investigated in vitro on tetratrichomonas gallinarum and histomonas meleagridis. on t. gallinarum, the minimal lethal concentration (mlc) at 24 hours was ...200312847923
parasites from farmed ostriches (struthio camelus) and rheas (rhea americana) in europe.during a 4-year-period, more than 500 ostriches and several rheas, all born in european countries and raised in spain and portugal, have been analyzed for the presence of ectoparasites and endoparasites. a total of 29 parasite species have been found, most of them of the gastrointestinal tract. some of the helminth species found may represent spureous parasitosis, as only the eggs (of an ascarid and a trematode) were found in some samples. from the organisms identified, the ectoparasites (lice-s ...200212072221
genetic differentiation and biochemical polymorphism among trichomonads.isoenzyme electrophoresis was used to study levels of genetic differentiation among strains and clones of trichomonas gallinae, trichomonas vaginalis, tritrichomonas foetus, tetratrichomonas gallinarum, and pentatrichomonas hominis. strain variation was found within t. gallinae, t. vaginalis, and t. foetus, however, levels of enzyme polymorphism were greater in t. gallinae than in t. vaginalis or t. foetus. isoenzyme genotypes were not a stable property of t. gallinae clones cultivated in vitro. ...19883418458
experimental infections in chickens with chilomastix gallinarum, tetratrichomonas gallinarum, and tritrichomonas eberthi.flagellates from the caeca of a diseased hen and a diseased goose were transmitted to 35 specific pathogen-free (spf) chickens. the flagellates of chicken origin were identified as chilomastix gallinarum, tritrichomonas eberthi, and tetratrichomonas gallinarum. t. eberthi was not detected in the material of goose origin. morphologic studies did not reveal any differences between chilomastix and tetratrichomonas specimens from chicken or goose origin. the species from the goose were identified as ...19911830966
[pathogenicity of tetratrichomonas gallinarum].eight axenic strains of tetratrichomonas gallinarum were isolated from the caeca or cloacae of ducks and geese. different growth curves were determined for each of the strains and the generation times were calculated, ranging from 2 h 36 min to 4 h 44 min. the axenic cultures of seven t. gallinarium strains were subjected to the honigberg test on inbred mice. the average volumes of the six-day-old subcutaneous abscesses ranged from 129.98 to 185.63 mm3. selected strains of trichomonads were used ...19911746100
axenization and optimization of in vitro growth of clonal cultures of tetratrichomonas gallinarum and trichomonas gallinae.a rapid and simple procedure was established to obtain clonal axenic cultures of tetratrichomonas gallinarum and trichomonas gallinae and to optimize their in vitro growth conditions. medium 199 was used for axenization of two genetically different clones of t. gallinarum and t. gallinae. six different media were used to optimize the growth behaviour of axenically grown parasites: medium 199, tym, tyi-s-33, hollander fluid (hf), trichomonas vaginalis (tv) and modified tv media. the highest cell ...201019766633
identification of a putatively novel trichomonad species in the intestine of a common quail (coturnix coturnix).a common quail (coturnix coturnix) from a private keeping died unexpectedly and showed a moderate lymphocytic infiltration of the colonic mucosa associated with numerous protozoa-like objects at the pathological examination. these organisms were further identified using chromogenic in situ hybridization (ish) and gene sequencing. ish was performed on paraffin embedded tissue sections and produced a positive signal using a probe specific for the 18s ribosomal rna (rrna) gene of the order trichomo ...201121839582
loop-mediated isothermal amplification assay for detection of histomonas meleagridis infection in chickens targeting the 18s rrna sequences.histomonas meleagridis is the causative agent of histomonosis, a disease of gallinaceous fowl characterized by necrotic typhlitis, hepatitis, and high mortality. to develop a rapid and sensitive method for specific detection of h. meleagridis, an assay based on loop-mediated isothermal amplification (lamp) targeting the 18s rrna gene was established. the detection limit of the lamp assay was 10 copies for standard plasmids containing an 18s rrna gene fragment, which was superior to that of a cla ...201424320623
aberrant clinical appearance and pathomorphology noticed during an outbreak of histomonosis indicates a different pathogenesis of histomonas meleagridis genotype 2.the present case report describes a remarkable feature of histomonas meleagridis characterized by aberrant clinical appearance and pathomorphologic lesions, which were mainly confined to the ceca, noticed during a field outbreak of histomonosis. in a flock of meat turkey toms, sudden death was noticed at the end of week 5 in the absence of specific clinical signs. instead of the well-known sulfur-colored feces, some caseous cores were found in the litter. mortality up to 17% per week was noticed ...201526478167
loop-mediated isothermal amplification assay for detection of histomonas meleagridis infection in chickens targeting the 18s rrna sequences.histomonas meleagridis is the causative agent of histomonosis, a disease of gallinaceous fowl characterized by necrotic typhlitis, hepatitis, and high mortality. to develop a rapid and sensitive method for specific detection of h. meleagridis, an assay based on loop-mediated isothermal amplification (lamp) targeting the 18s rrna gene was established. the detection limit of the lamp assay was 10 copies for standard plasmids containing an 18s rrna gene fragment, which was superior to that of a cla ...201324325302
actin cytoskeleton demonstration in trichomonas vaginalis and in other trichomonads.the flagellate form of trichomonas vaginalis (t v) transforms to amoeboid cells upon adherence to converslips. they grow and their nuclei divide without undergoing cytokinesis, yielding giant cells and a monolayer of t v f-actin was demonstrated in trichomonas vaginalis by fluorescence microscopy using phalloidin and an anti-actin mab which labelled the cytoplasm of both the flagellate and amoeboid forms. comparative electrophoresis and immunoblotting established that the actin band has the same ...19969175265
experimental infection of turkeys and chickens with a clonal strain of tetratrichomonas gallinarum induces a latent infection in the absence of clinical signs and lesions.the pathogenicity of a mono-eukaryotic culture of tetratrichomonas gallinarum in specific pathogen free (spf) chickens and turkeys was studied. two experiments of identical design were performed: the first with spf chickens and the second with commercial turkeys. each experiment included three groups. groups 1 and 2 each contained 12 infected and three in-contact birds. the birds in these groups were infected on the first day of life, either cloacally (group 1) or orally (group 2). group 3 consi ...201120708742
detection of dna of histomonas meleagridis and tetratrichomonas gallinarum in german poultry flocks between 2004 and 2008.between 2004 and 2008, 338 samples from 156 german turkey, chicken, and peacock flocks with suspected histomonosis (histomoniasis) were sent to the institute for poultry diseases of the free university berlin. most samples were from ceca or livers; the other samples were organ pools or were taken from other organs or the environment. in 108 samples from 65 flocks, histomonal dna was detected by polymerase chain reaction (pcr). tetratrichomonas gallinarum dna was found in 5.3% of investigated sam ...201020945783
the cytoskeleton of parabasalian parasites comprises proteins that share properties common to intermediate filament proteins.certain protist lineages bear cytoskeletal structures that are germane to them and define their individual group. trichomonadida are excavate parasites united by a unique cytoskeletal framework, which includes tubulin-based structures such as the pelta and axostyle, but also other filaments such as the striated costa whose protein composition remains unknown. we determined the proteome of the detergent-resistant cytoskeleton of tetratrichomonas gallinarum. 203 proteins with homology to trichomon ...201627744090
Trichomonas gallinae, in comparison to Tetratrichomonas gallinarum, induces distinctive cytopathogenic effects in tissue cultures.In the present study the interaction of three genetically different clonal cultures of Trichomonas gallinae and Tetratrichomonas gallinarum with a permanent chicken liver (LMH) and a permanent quail fibroblast (QT35) cell culture was studied. Proliferation of T. gallinae cells was associated with a disintegration of the cell monolayer. The initial lesions on the LMH monolayer consisted of a progressive accumulation of the flagellate, forming clumps attached to the monolayer. A prolonged incubati ...201122172581
coligranuloma (hjärre and wramby's disease) reconsidered.coligranulomatosis (hjärre and wramby's disease) is considered to be a disease of chickens, turkeys and partridges that occurs sporadically in individual, adult birds. therefore, the condition is not of economic importance, but is of interest due to the similarity of its lesions to those of tuberculosis. in a number of cases the disease could be reproduced by inoculation via artificial routes of granuloma homogenate or escherichia coli bacteria isolated from the lesions. oral inoculations always ...201728277781
a single strain of tetratrichomonas gallinarum causes fatal typhlohepatitis in red-legged partridges (alectoris rufa) to be distinguished from histomonosis.typhlohepatitis was observed in a flock of 2500 red-legged partridges in great britain, characterized by the sudden deaths of 15 birds within 2 days. necropsy of five dead birds revealed severe lesions in the caeca with thickened caecal walls, a reddened lining and bloody contents. the livers contained multiple miliary lesions and similar pathological changes were found in the spleens of some birds. microscopic examination of intestinal contents showed the occurrence of coccidial oocysts in two ...201425175532
cryptic species within the tetratrichomonas gallinarum species complex revealed by molecular polymorphism.tetratrichomonas gallinarum is a widespread intestinal parasite of galliform and anseriform birds. the pathogenicity of this species is controversial, presenting an unsettled problem as yet. we analysed the polymorphism and genetic relationship among 29 isolates of t. gallinarum obtained from eight bird species and five t. gallinarum-like isolates from the oral cavity and lower respiratory tract of human patients. two methods were used for the analyses: rapd and sequencing of 16s rrna, 5.8s rrna ...200415725528
trichomonads in birds--a review.members of the family trichomonadidae, mainly trichomonas gallinae and tetratrichomonas gallinarum, represent important parasites in birds with worldwide presence, since being reported in the 19th century. especially columbiformes, falconiformes and strigiformes can be severely affected by trichomonads, whereas the majority of infections in galliformes and anatiformes are subclinical although severe infections are occasionally reported. with the recent appearance of deadly infections in wild pas ...201424476968
hepatitis and splenitis due to systemic tetratrichomoniasis in an american white pelican (pelecanus erythrorhynchos).a free-ranging, young adult, female american white pelican (pelecanus erythrorhynchos), found dead on the grounds of the san diego zoo safari park in southern california, had severe multifocal to coalescing necrotizing hepatitis and splenitis on postmortem examination. histologically, within the large areas of necrosis were myriad pleomorphic, 5-20 µm in diameter, protozoal organisms with 1 to multiple nuclei. ultrastructurally, the organisms were consistent with a trichomonad flagellate. polyme ...201323674462
fatal hepatic tetratrichomoniasis in a juvenile waldrapp ibis (geronticus eremita).waldrapp ibis (geronticus eremita) are a critically endangered species, and there are currently more birds in captivity than in the wild. a juvenile, male waldrapp ibis housed in a mixed-species exhibit was found dead with no premonitory signs. necropsy revealed extensive necrotizing hepatitis associated with numerous pleomorphic protozoa that were immunohistochemically reactive with antibodies raised against tritrichomonas foetus, a parasite of cattle. electron microscopy confirmed the organism ...201323404476
pathologic and molecular characterization of histomoniasis in peafowl ( pavo cristatus).histomonas meleagridis is a flagellate protozoan organism that can cause severe necrotizing typhlitis and hepatitis in gallinaceous birds. peafowl ( pavo spp.) have been shown to be susceptible to histomoniasis in experimental settings, but there are few reports of natural histomoniasis in this species. a retrospective study of the archived cases at 2 veterinary diagnostic laboratories in the united states yielded 5 cases of peafowl with gross and histologic findings characteristic of histomonia ...201728065124
detection and quantification of histomonas meleagridis by real-time pcr targeting single copy genes.histomonas meleagridis, a protozoan parasite that can infect gallinaceous birds, affects mainly the liver and caeca of infected birds. as a consequence of the recent ban of chemotherapeuticals in europe and the usa, histomonosis gained somewhat more attention due to its re-emergence and the fact that there is no effective treatment available. therefore, special attention is now also given towards the diagnosis and the control of the disease. in the actual study we report the development of highl ...201526319200
establishing mono-eukaryotic histomonas meleagridis cultures from in vivo infection contaminated with tetratrichomonas gallinarum and blastocystis spp.summary the necessity to easily establish histomonas meleagridis cultures has been underlined extensively by many researchers in order to gain more insights in the biology of h. meleagridis. in addition the occurrence of different protozoa in the caeca of birds impedes, however, the isolation and propagation of h. meleagridis from field outbreaks. therefore, in a kinetic study using transmission electron microscopy the deleterious effects of adventitious protozoa including tetratrichomonas galli ...201323790160
molecular characterization of trichomonads isolated from animal hosts in the philippines.trichomonads are amitochondrial anaerobic flagellated protists that are either parasites or commensals, generally living in the digestive or genitourinary tract of humans and animals. it has been reported that these protozoa can migrate to other sites in their target host, can adapt to new hosts, and are capable of zoonotic transmission. in this study, 59 trichomonad isolates from different animal hosts in the philippines were identified and characterized. primer sets were designed and were succ ...201323622817
granuloma disease in flocks of productive layers caused by tetratrichomonas gallinarum.in 2013, seven outbreaks of granuloma disease occurred in dutch flocks of productive layers housed on different farms. these outbreaks were characterized by increased mortality and high incidence of granulomas, mainly in caeca (340/408 hens = 83%) and livers (69/408 hens = 17%). mortality started to increase between 21 and 35 weeks of age and reached 3.7% to 11.0% exceeding the breeder's norm in periods ranging from 9 to 48 weeks. some flocks also showed decreased egg production and/or loss of m ...201626950827
phylogenetic position of parabasalid symbionts from the termite calotermes flavicollis based on small subunit rrna sequences.small subunit rdna genes were amplified by polymerase chain reaction using specific primers from mixed-population dna obtained from the whole hindgut of the termite calotermes flavicollis. comparative sequence analysis of the clones revealed two kinds of sequences that were both from parabasalid symbionts. in a molecular tree inferred by distance, parsimony and likelihood methods, and including 27 parabasalid sequences retrieved from the data bases, the sequences of the group ii (clones cf5 and ...200011032309
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