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1.
Protist ; 173(4): 125883, 2022 08.
Artigo em Inglês | MEDLINE | ID: mdl-35660751

RESUMO

The vast majority of the more than 450 described species of Parabasalia are intestinal symbionts or parasites of animals. This endobiotic life-history is presumably ancestral although the root of Parabasalia still needs to be robustly established. The half-dozen putatively free-living species thus far described are likely independently derived from endobiotic ancestors and represent the most neglected ecological group of parabasalids. Thus, we isolated and cultivated 45 free-living strains of Parabasalia obtained from a wide variety of anoxic sediments to conduct detailed morphological and SSU rRNA gene phylogenetic analyses. Sixteen species of trichomonads were recovered. Among them, we described seven new species, three new genera, two new families, and one new order. Most of the newly described species were more or less closely related to members of already described genera. However, we uncovered a new deep-branching lineage without affinity to any currently known group of Parabasalia. The newly discovered free-living parabasalids will be key taxa in comparative analyses aimed at rooting the entire lineage and deciphering the evolutionary innovations involved in transitioning between endobiotic and free-living habitats.


Assuntos
Parabasalídeos , Parasitos , Animais , Evolução Biológica , Parabasalídeos/genética , Filogenia
3.
Acta Parasitol ; 67(1): 535-538, 2022 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-34677797

RESUMO

INTRODUCTION: Lophomoniasis is caused by Lophomonas spp., a new emerging protozoan, which commonly affects the human lower respiratory tract. The Lophomonas parasite mostly lives commensally in the hindgut of cockroaches. CASE PRESENTATION: We present the case of a 33-year-old woman, 30 weeks pregnant, who had severe COVID-19. She was intubated upon admission and began the routine COVID-19 treatment. To rule out possible super infection dual with COVID-19, microscopic examination of the patient's mini-bronchoalveolar lavage (mini-BAL) specimen, revealed L. blattarum, which was identified by the SSU rRNA-PCR and sequencing approaches (accession number: MZ093069). According to that, the patient was treated successfully with metronidazole. CONCLUSION: To prevent serious complications, lophomoniasis should be listed in co-morbidity cases of COVID-19 infection during the COVID-19 pandemic worldwide. To the best of our knowledge, this is the first co-infection of Lophomonas blattarum and COVID-19 in the world which has been confirmed using a molecular approach.


Assuntos
COVID-19 , Parabasalídeos , Adulto , COVID-19/epidemiologia , Feminino , Humanos , Morbidade , Pandemias
4.
Protist ; 172(5-6): 125836, 2021.
Artigo em Inglês | MEDLINE | ID: mdl-34757297

RESUMO

Microjoenia are obligate symbionts of termites. The genus was erected in 1892 for small cells with many flagella that insert near, but not directly from, the cell apex, and an axostyle that can protrude from the cell posterior. Although ultrastructural studies have been carried out on three Microjoenia species to date, no molecular data have been directly attributed to any species. Microjoenia are classified within the parabasalian class Spirotrichonymphea, which is characterized by flagellar bands that emerge near the cell apex and proceed posteriorly in a right-handed helix. In Microjoenia, however, the flagellar bands are very short and proceed longitudinally or with a weakly observable helix. In this study, we have amplified and sequenced the 18S ribosomal RNA gene from individually isolated Microjoenia cells from Reticulitermes and Hodotermopsis hosts as part of an ongoing effort to understand the phylogeny of Spirotrichonymphea and their coevolution with termites. In our 18S rRNA gene phylogeny, Microjoenia forms the sister lineage to Spirotrichonympha, though many other evolutionary relationships within Spirotrichonymphea remain unresolved.


Assuntos
Isópteros , Parabasalídeos , Animais , Parabasalídeos/genética , Filogenia , RNA Ribossômico 18S/genética , Simbiose
5.
Acta Parasitol ; 66(4): 1510-1516, 2021 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-34115281

RESUMO

BACKGROUND: In the last decade, several cases of bronchopulmonary lophomoniasis (BPL) have been recorded. Little information is available about epidemiological aspects on Lophomonas infection among BPL patients. The present study was aimed to investigate the prevalence of Lophomonas spp. infection in patients who were referred to the Iranian National Registry Center for Lophomoniasis (INRCL), using morphological and molecular tests. SUBJECTS AND METHODS: We examined patients enrolled in the INRCL from 2017 to 2019 at the Mazandaran University of Medical Sciences, northern Iran. All bronchoalveolar lavage fluid (BALF) and two nasal discharges of the patients were examined by both microscopic and small-subunit ribosomal RNA (SSU rRNA) PCR methods. To confirm the species of Lophomonas, two positive samples were sequenced. RESULTS: In this study, 321 specimens (including 319 BALF and 2 nasal discharges) were microscopically examined. Lophomonas spp. was found in 45(14%) (n = 44 BAL; n = 1 nasal discharge). The mean age of infected patients was 54.9 ± 17.1 years. The following morphological characteristics were observed in both fresh and Papanicolaou-stained smears to identify Lophomonas spp. All microscopically positive specimens were confirmed with genus-specific PCR technique. The obtained sequences were deposited in Gen Bank under the accession numbers (MN243135-36). The BLAST analysis of our two sequences with the only available sequence in the Gen Bank of the Thailand strain of L. blattarum, showed identity of 99-100% and 98.51%, respectively. CONCLUSION: To the best of our knowledge, this is the first registry-based study regarding lophomoniasis worldwide. According to our study, the conventional PCR test is an available and reliable tool for confirming the Lophomonas parasite in clinical samples. Moreover, the results confirmed that L. blattarum is circulating at least in our region.


Assuntos
Parabasalídeos , Adulto , Idoso , Humanos , Irã (Geográfico)/epidemiologia , Pessoa de Meia-Idade , Técnicas de Amplificação de Ácido Nucleico , Prevalência , Sistema de Registros
6.
Sci Rep ; 11(1): 7270, 2021 03 31.
Artigo em Inglês | MEDLINE | ID: mdl-33790354

RESUMO

Cristamonadea is a large class of parabasalian protists that reside in the hindguts of wood-feeding insects, where they play an essential role in the digestion of lignocellulose. This group of symbionts boasts an impressive array of complex morphological characteristics, many of which have evolved multiple times independently. However, their diversity is understudied and molecular data remain scarce. Here we describe seven new species of cristamonad symbionts from Comatermes, Calcaritermes, and Rugitermes termites from Peru and Ecuador. To classify these new species, we examined cells by light and scanning electron microscopy, sequenced the symbiont small subunit ribosomal RNA (rRNA) genes, and carried out barcoding of the mitochondrial large subunit rRNA gene of the hosts to confirm host identification. Based on these data, five of the symbionts characterized here represent new species within described genera: Devescovina sapara n. sp., Devescovina aymara n. sp., Macrotrichomonas ashaninka n. sp., Macrotrichomonas secoya n. sp., and Macrotrichomonas yanesha n. sp. Additionally, two symbionts with overall morphological characteristics similar to the poorly-studied and probably polyphyletic 'joeniid' Parabasalia are classified in a new genus Runanympha n. gen.: Runanympha illapa n. sp., and Runanympha pacha n. sp.


Assuntos
Isópteros , Parabasalídeos , Simbiose , Animais , Parabasalídeos/classificação , Parabasalídeos/fisiologia
8.
9.
Eur J Protistol ; 76: 125742, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-33161208

RESUMO

The desert dampwood termite Paraneotermes simplicicornis harbors several species of obligately symbiotic protists that support its nutrition by fermenting lignocellulose. Among them are three morphotypes with the dexiotropic spiraling flagellar bands characteristic of Spirotrichonymphea (Parabasalia). The largest morphotype, characterized by an elongated cell apex with axial columella and internally positioned spiraling flagellar bands, was previously described as Spirotrichonympha polygyra. A smaller morphotype, with similarly internalized flagellar bands but a more rounded posterior without a protruding axostyle, was previously reported but not named. The smallest morphotype has surface flagellar bands and can attach to other protist cells by its apex. In this study, we combine light microscopy of live specimens and 18S rRNA gene sequencing of individually isolated cells to better understand the diversity of symbionts in P. simplicicornis. We found that S. polygyra branches distantly from true Spirotrichonympha, which are associated with Reticulitermes termites. Thus, we propose the new genus Cuppa to accommodate C. polygyra n. comb. (type species) and the similar but smaller morphotype Cuppa taenia n. sp. The undescribed smallest morphotype can be excluded from all previously described Spirotrichonymphea genera by molecular and behavioral evidence, so we propose Fraterculus simplicicornis n. gen., n. sp., to accommodate this organism.


Assuntos
Isópteros/parasitologia , Parabasalídeos/classificação , Parabasalídeos/fisiologia , Simbiose , Animais , DNA de Protozoário/genética , Parabasalídeos/genética , Filogenia , RNA Ribossômico 18S/genética , Especificidade da Espécie
10.
EMBO J ; 39(22): e106249, 2020 11 16.
Artigo em Inglês | MEDLINE | ID: mdl-32954505

RESUMO

Centrioles are polarized microtubule-based organelles that seed the formation of cilia, and which assemble from a cartwheel containing stacked ring oligomers of SAS-6 proteins. A cryo-tomography map of centrioles from the termite flagellate Trichonympha spp. was obtained previously, but higher resolution analysis is likely to reveal novel features. Using sub-tomogram averaging (STA) in T. spp. and Trichonympha agilis, we delineate the architecture of centriolar microtubules, pinhead, and A-C linker. Moreover, we report ~25 Å resolution maps of the central cartwheel, revealing notably polarized cartwheel inner densities (CID). Furthermore, STA of centrioles from the distant flagellate Teranympha mirabilis uncovers similar cartwheel architecture and a distinct filamentous CID. Fitting the CrSAS-6 crystal structure into the flagellate maps and analyzing cartwheels generated in vitro indicate that SAS-6 rings can directly stack onto one another in two alternating configurations: with a slight rotational offset and in register. Overall, improved STA maps in three flagellates enabled us to unravel novel architectural features, including of centriole polarity and cartwheel stacking, thus setting the stage for an accelerated elucidation of underlying assembly mechanisms.


Assuntos
Centríolos/ultraestrutura , Microscopia Crioeletrônica/métodos , Tomografia/métodos , Adesão Celular , Cílios/ultraestrutura , Microtúbulos/ultraestrutura , Parabasalídeos/citologia
11.
J Eukaryot Microbiol ; 67(6): 626-641, 2020 11.
Artigo em Inglês | MEDLINE | ID: mdl-32603489

RESUMO

Coptotermes formosanus Shiraki and Coptotermes gestroi (Wasmann) (Blattoidea: Rhinotermitidae) are invasive subterranean termite pest species with a major global economic impact. However, the descriptions of the mutualistic protist communities harbored in their respective hindguts remain fragmentary. The C. formosanus hindgut has long been considered to harbor three protist species, Pseudotrichonympha grassii (Trichonymphida), Holomastigotoides hartmanni, and Cononympha (Spirotrichonympha) leidyi (Spirotrichonymphida), but molecular data have suggested that the diversity may be higher. Meanwhile, the C. gestroi community remains undescribed except for Pseudotrichonympha leei. To complete the characterization of these communities, hindguts of workers from both termite species were investigated using single-cell PCR, microscopy, cell counts, and 18S rRNA amplicon sequencing. The two hosts were found to harbor intriguingly parallel protist communities, each consisting of one Pseudotrichonympha species, two Holomastigotoides species, and two Cononympha species. All protist species were unique to their respective hosts, which last shared a common ancestor ~18 MYA. The relative abundances of protist species in each hindgut differed remarkably between cell count data and 18S rRNA profiles, calling for caution in interpreting species abundances from amplicon data. This study will enable future research in C. formosanus and C. gestroi hybrids, which provide a unique opportunity to study protist community inheritance, compatibility, and potential contribution to hybrid vigor.


Assuntos
Sistema Digestório/parasitologia , Isópteros/parasitologia , Parabasalídeos/classificação , Parabasalídeos/genética , Animais , DNA de Protozoário/genética , Interações Hospedeiro-Parasita , Filogenia , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Análise de Célula Única , Simbiose
12.
Acta Parasitol ; 65(4): 831-836, 2020 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-32472401

RESUMO

PURPOSE: The aim of this study was to investigate protozoan cysts and oocyts in the faecal pellets of the German cockroach (Blattella germanica), with emphasis on the prevalence of Lophomonas blattarum. METHODS: Thirty adult Blattella germanica (12 males; 18 females) were trapped near Madrid, Spain. Expelled faecal pellets were collected in the laboratory over 5 days in two plastic containers. Protozoan cysts from one container were weighted and those in the other used for morphologically identification. RESULTS: Protozoan cysts/oocysts per gram of faecal pellet were: Nyctotherus sp. (0.0019/g), Entamoeba (0.0007/g), Balantidium coli (0.0001/g), Lophomonas blattarum (0.00038/g). Observation of 189 protozoan cysts/oocysts indicated the following prevalence: Nyctotherus sp. 65 (34.4%); Gregarina spp. 34 (18%); Entamoeba sp. 24 (12.7%); Cryptosporidium sp. 17 (9%); Coccidia 16 (8.4%); Lophomonas blattarum 13 (6.8%); Balantidium coli 4 (2.1%); and unclassified 16 (8.4%). CONCLUSION: Lophomonas blattarum has previously been demonstrated in the gut of cockroaches, but this is the first assessment of the prevalence in Blattella germanica in faecal pellets. The presence of protozoa in faecal pellets provides some evidence for one step in a hypothesised route of respiratory infection whereby protozoa from household insects enter the respiratory tract.


Assuntos
Blattellidae , Baratas , Criptosporidiose , Cryptosporidium , Cistos , Parabasalídeos , Adulto , Animais , Feminino , Humanos , Masculino
14.
J Eukaryot Microbiol ; 67(2): 268-272, 2020 03.
Artigo em Inglês | MEDLINE | ID: mdl-31560813

RESUMO

Hoplonympha natator is an obligate symbiont of Paraneotermes simplicicornis (Kalotermitidae), from southwestern North America. Another Hoplonympha species inhabits Hodotermopsis sjostedti (Archotermopsidae), from montane Southeast Asia. The large phylogenetic and geographical distance between the hosts makes the distribution of Hoplonympha puzzling. Here, we report the phylogenetic position of H. natator from P. simplicicornis through maximum likelihood and Bayesian analysis of 18S rRNA genes. The two Hoplonympha species form a clade with a deep node, making a recent symbiont transfer unlikely. The distribution of Hoplonympha may be due to an ancient transfer or strict vertical inheritance with differential loss from other hosts.


Assuntos
Isópteros/parasitologia , Parabasalídeos/classificação , Animais , Arizona , Teorema de Bayes , Parabasalídeos/genética , Filogenia , RNA de Protozoário/análise , RNA Ribossômico 18S/análise , Especificidade da Espécie , Simbiose
16.
BMC Evol Biol ; 19(1): 162, 2019 08 02.
Artigo em Inglês | MEDLINE | ID: mdl-31375061

RESUMO

BACKGROUND: Two spliceosomal intron types co-exist in eukaryotic precursor mRNAs and are excised by distinct U2-dependent and U12-dependent spliceosomes. In the diplomonad Giardia lamblia, small nuclear (sn) RNAs show hybrid characteristics of U2- and U12-dependent spliceosomal snRNAs and 5 of 11 identified remaining spliceosomal introns are trans-spliced. It is unknown whether unusual intron and spliceosome features are conserved in other diplomonads. RESULTS: We have identified spliceosomal introns, snRNAs and proteins from two additional diplomonads for which genome information is currently available, Spironucleus vortens and Spironucleus salmonicida, as well as relatives, including 6 verified cis-spliceosomal introns in S. vortens. Intron splicing signals are mostly conserved between the Spironucleus species and G. lamblia. Similar to 'long' G. lamblia introns, RNA secondary structural potential is evident for 'long' (> 50 nt) Spironucleus introns as well as introns identified in the parabasalid Trichomonas vaginalis. Base pairing within these introns is predicted to constrain spatial distances between splice junctions to similar distances seen in the shorter and uniformly-sized introns in these organisms. We find that several remaining Spironucleus spliceosomal introns are ancient. We identified a candidate U2 snRNA from S. vortens, and U2 and U5 snRNAs in S. salmonicida; cumulatively, illustrating significant snRNA differences within some diplomonads. Finally, we studied spliceosomal protein complements and find protein sets in Giardia, Spironucleus and Trepomonas sp. PC1 highly- reduced but well conserved across the clade, with between 44 and 62 out of 174 studied spliceosomal proteins detectable. Comparison with more distant relatives revealed a highly nested pattern, with the more intron-rich fornicate Kipferlia bialata retaining 87 total proteins including nearly all those observed in the diplomonad representatives, and the oxymonad Monocercomonoides retaining 115 total proteins including nearly all those observed in K. bialata. CONCLUSIONS: Comparisons in diplomonad representatives and species of other closely-related metamonad groups indicates similar patterns of intron structural conservation and spliceosomal protein composition but significant divergence of snRNA structure in genomically-reduced species. Relative to other eukaryotes, loss of evolutionarily-conserved snRNA domains and common sets of spliceosomal proteins point to a more streamlined splicing mechanism, where intron sequences and structures may be functionally compensating for the minimalization of spliceosome components.


Assuntos
Sequência Conservada , Diplomonadida/genética , Íntrons/genética , Parabasalídeos/genética , Filogenia , Spliceossomos/genética , Regiões 5' não Traduzidas/genética , Pareamento de Bases/genética , Sequência de Bases , Genoma , Conformação de Ácido Nucleico , Splicing de RNA/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Nuclear Pequeno/química , RNA Nuclear Pequeno/genética , Proteínas Ribossômicas/genética
17.
Acta Parasitol ; 64(2): 390-393, 2019 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-31165985

RESUMO

INTRODUCTION: We report a case of lophomoniasis in an immunocompetent patient with acute paranasal sinusitis from the north of Iran whose disease was diagnosed by both microscopic and molecular methods. CASE PRESENTATION: The patient, a 40-year-old woman, suffered from upper respiratory infection, was referred to the Iranian National Registry Center for Lophomoniasis (INRCL) at the Mazandaran University of Medical Sciences, Sari, Iran, for diagnosis. A direct wet mount of nasal discharge revealed the flagellate protozoa morphologically identified Lophomonas blattarum. Moreover, through a specific polymerase chain reaction (PCR) of nasal discharge, a 214-bp band was observed, confirming the genus Lophomonas spp. The patient was treated successfully with metronidazole 500 mg t.i.d for 1 week. CONCLUSION: To the best of our knowledge, this is the first molecular detection of lophomoniasis in the literature. According to our preliminary study, a reliable PCR test is available now for detecting the Lophomonas parasite.


Assuntos
Pneumopatias Parasitárias/diagnóstico , Parabasalídeos/isolamento & purificação , Adulto , Antiprotozoários/uso terapêutico , Feminino , Humanos , Imunocompetência , Irã (Geográfico) , Pneumopatias Parasitárias/parasitologia , Metronidazol/uso terapêutico , Técnicas de Diagnóstico Molecular , Parabasalídeos/genética , Sinusite/parasitologia
18.
J Eukaryot Microbiol ; 66(6): 882-891, 2019 11.
Artigo em Inglês | MEDLINE | ID: mdl-31033101

RESUMO

Holomastigotes is a protist genus (Parabasalia: Spirotrichonymphea) that resides in the hindguts of "lower" termites. It can be distinguished from other parabasalids by spiral flagellar bands that run along the entire length of the cell, an anterior nucleus, a reduced or absent axostyle, the presence of spherical vesicles inside the cells, and the absence of ingested wood particles. Eight species have been described based on their morphology so far, although no molecular data were available prior to this study. We determined the 18S rRNA gene sequences of Holomastigotes from the hindguts of Hodotermopsis sjostedti, Reticulitermes flavipes, Reticulitermes lucifugus, and Reticulitermes tibialis. Phylogenetic analyses placed all sequences in an exclusive and well-supported clade with the type species, Holomastigotes elongatum from R. lucifugus. However, the phylogenetic position of Holomastigotes within the Spirotrichonymphea was not resolved. We describe two new species, Holomastigotes flavipes n. sp. and Holomastigotes tibialis n. sp., inhabiting the hindguts of R. flavipes and R. tibialis, respectively.


Assuntos
Isópteros/parasitologia , Parabasalídeos/classificação , Animais , Sistema Digestório/parasitologia , Parabasalídeos/citologia , Parabasalídeos/genética , Filogenia , RNA de Protozoário/análise , RNA Ribossômico 18S/análise , Simbiose
19.
Turkiye Parazitol Derg ; 43(1): 44-46, 2019 Mar 28.
Artigo em Inglês | MEDLINE | ID: mdl-30938137

RESUMO

The frequency of bronchopulmonary protozoan infections has raised due to increased number of immunosuppressed patients in recent years. One of them is Lophomonas blattarum which is a multi-flagellated protozoan parasite of termites and several cockroach species. The drug regimens commonly used in bronchopulmonary infections are not effective against L. blattarum. Therefore, rapid and accurate diagnosis of L. blattarum infection is of great importance in the treatment success. The laboratory diagnosis of L. blattarum infection is made on the basis of observation of the characteristic trophozoite in various samples. It is of a great importance to distinguish the protozoon from ciliated respiratory epithelium to avoid wrong positivity. The presented case developed an acute respiratory distress syndrome a short while after taking nivolumab immunotherapy. The morphological features of L. blattarum were demonstrated by examining the bronchoalveolar lavage fluid of the patient under light microscopy. Additionally, URL (https://youtu.be/EQIAsFl6AJY) of a smart-phone based video of trophozoite of this patient was added into this report.


Assuntos
Pneumopatias Parasitárias/diagnóstico , Parabasalídeos/isolamento & purificação , Infecções por Protozoários/diagnóstico , Trofozoítos/isolamento & purificação , Idoso , Líquido da Lavagem Broncoalveolar , Diagnóstico Diferencial , Humanos , Hospedeiro Imunocomprometido , Imunoterapia , Pneumopatias Parasitárias/parasitologia , Masculino , Infecções por Protozoários/parasitologia , Smartphone , Gravação em Vídeo
20.
PLoS One ; 14(2): e0212429, 2019.
Artigo em Inglês | MEDLINE | ID: mdl-30807611

RESUMO

The exoproteome of parasitic protists constitutes extracellular proteins that play a fundamental role in host-parasite interactions. Lytic factors, especially secreted proteases, are capable of modulating tissue invasion, thereby aggravating host susceptibility. Despite the important role of exoproteins during infection, the exoproteomic data on Histomonas meleagridis are non-existent. The present study employed traditional 1D-in-gel-zymography (1D-IGZ) and micro-LC-ESI-MS/MS (shotgun proteomics), to investigate H. meleagridis exoproteomes, obtained from a clonal virulent and an attenuated strain. Both strains were maintained as mono-eukaryotic monoxenic cultures with Escherichia coli. We demonstrated active in vitro secretion kinetics of proteases by both parasite strains, with a widespread proteolytic activity ranging from 17 kDa to 120 kDa. Based on protease inhibitor susceptibility assay, the majority of proteases present in both exoproteomes belonged to the family of cysteine proteases and showed stronger activity in the exoproteome of a virulent H. meleagridis. Shotgun proteomics, aided by customized database search, identified 176 proteins including actin, potential moonlighting glycolytic enzymes, lytic molecules such as pore-forming proteins (PFPs) and proteases like cathepsin-L like cysteine protease. To quantify the exoproteomic differences between the virulent and the attenuated H. meleagridis cultures, a sequential window acquisition of all theoretical spectra mass spectrometric (SWATH-MS) approach was applied. Surprisingly, results showed most of the exoproteomic differences to be of bacterial origin, especially targeting metabolism and locomotion. By deciphering such molecular signatures, novel insights into a complex in vitro protozoan- bacteria relationship were elucidated.


Assuntos
Parabasalídeos/genética , Parabasalídeos/microbiologia , Animais , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Cisteína Proteases/genética , Cisteína Proteases/metabolismo , Exopeptidases/genética , Exopeptidases/metabolismo , Interações Hospedeiro-Parasita/genética , Parabasalídeos/patogenicidade , Aves Domésticas , Doenças das Aves Domésticas/microbiologia , Doenças das Aves Domésticas/parasitologia , Mapas de Interação de Proteínas , Proteoma/genética , Proteômica , Infecções Protozoárias em Animais/microbiologia , Infecções Protozoárias em Animais/parasitologia , Proteínas de Protozoários/genética , Proteínas de Protozoários/metabolismo , Virulência/genética
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