RESUMO
The tandemly arranged CPB genes of Leishmania mexicana are polycistronically transcribed and encode cysteine proteases that are differentially stage-specific; CPB1 and CPB2 are expressed predominantly in metacyclics, whereas CPB3-CPB18 are expressed mainly in amastigotes. The mechanisms responsible for this differential expression have been studied via gene analysis and re-integration of individual CPB genes, and variants thereof, into a CPB-deficient parasite mutant. Comparison of the nucleotide sequences of the repeat units of CPB1 and CPB2 with CPB2.8 (typical of CPB3-CPB18) revealed two major regions of divergence as follows: one of 258 base pairs (bp) corresponding to the C-terminal extension of CPB2.8; another, designated InS, of 120 bp, with insertions totaling 57 bp, localized to the intercistronic region downstream of CPB1 and CPB2. Cell lines expressing CPB2.8 or CPB2 with the 3'-untranslated region and intercistronic sequence of CPB2.8 showed up-regulation in amastigotes. Conversely, metacyclic-specific expression occurred with CPB2 or CPB2.8 with the 3'-untranslated region and intercistronic sequence of CPB2. Moreover, the InS down-regulated expression in amastigotes of a reporter gene integrated into the CPB locus. It is proposed that the InS mediates metacyclic-specific stage-regulated expression of CPB by affecting the maturation of polycistronic pre-mRNA. This is the first well defined cis-regulatory element implicated in post-transcriptional stage-specific gene expression in Leishmania.
Assuntos
Cisteína Endopeptidases/biossíntese , Cisteína Endopeptidases/genética , Leishmania mexicana/enzimologia , Regiões 3' não Traduzidas , Animais , Sequência de Bases , Northern Blotting , Southern Blotting , Cloranfenicol O-Acetiltransferase/metabolismo , Cisteína Endopeptidases/química , Regulação para Baixo , Gelatina/química , Regulação da Expressão Gênica , Genes , Genes Reporter , Modelos Genéticos , Dados de Sequência Molecular , Mutação , Plasmídeos/metabolismo , Poli A/metabolismo , Estrutura Terciária de Proteína , Processamento Pós-Transcricional do RNA , RNA Mensageiro/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Fatores de Tempo , TransfecçãoRESUMO
An undescribed species of Haematoloechus inhabits the lungs of Rana vaillanti in northwestern Costa Rica. The new species is most similar morphologically to H. medioplexus, having a very small, but well-developed, ventral sucker and lacking extracecal uterine loops, and apparently was mis-identified previously as H. medioplexus in Rana palmipes from Colombia. It differs from H. medioplexus, notably by (1) the shape of the oral sucker, which is elliptical in H. meridionalis and spherical in H. medioplexus; (2) the posterior extent of the vitelline fields, which terminate at the same level in H. medioplexus, but always at different levels in H. meridionalis; (3) the position of the testes, which are immediately posterior to the seminal receptacle and are close together in the new species and far from the seminal receptacle and separated from each other in H. medioplexus; and (4) the location of the genital pore, which is ventral to the cecal bifurcation in the new species and ventral to the pharynx in H. medioplexus. In addition, the new species is unique among members of Haematoloechus by having flaps of tissue on the interior margins of the ventral sucker. The new species shows almost 5% sequence divergence from H. medioplexus in the variable D1 region of the 28s rDNA, complementing the morphological differences.
Assuntos
Pulmão/parasitologia , Ranidae/parasitologia , Trematódeos/classificação , Infecções por Trematódeos/veterinária , Animais , Sequência de Bases , Costa Rica , Dados de Sequência Molecular , Homologia de Sequência do Ácido Nucleico , Trematódeos/anatomia & histologiaRESUMO
Halipegus eschi n. sp. is described from the esophagus of Rana vaillanti from Guanacaste Province, Costa Rica. The new species differs from other known species of Halipegus on the basis of relative testis size, lateral extent of the uterus, vitelline follicle arrangement, egg size, and polar filament length.
Assuntos
Ranidae/parasitologia , Trematódeos/classificação , Infecções por Trematódeos/veterinária , Animais , Costa Rica , Esôfago/parasitologia , Trematódeos/anatomia & histologia , Infecções por Trematódeos/parasitologiaRESUMO
Removal of the pro-domain of a cysteine protease is essential for activation of the enzyme. We have engineered a cysteine protease (CPB2.8) of the protozoan parasite Leishmania mexicana by site-directed mutagenesis to remove the active site cysteine (to produce CPB(C25G)). When CPB(C25G) was expressed in a L. mexicana mutant lacking all CPB genes, the inactive pro-enzyme was processed to the mature protein and trafficked to the lysosome. These results show that auto-activation is not required for correct processing of CPB in vivo. When CPB(C25G) was expressed in a L. mexicana mutant lacking both CPA and CPB genes, the majority of the pro-enzyme remained unprocessed and accumulated in the flagellar pocket. These data reveal that CPA can directly or indirectly process CPB(C25G) and suggest that cysteine proteases are targeted to lysosomes via the flagellar pocket. Moreover, they show that another protease can process CPB in the absence of either CPA or CPB, albeit less efficiently. Abolition of the glycosylation site in the mature domain of CPB did not affect enzyme processing, targeting or in vitro activity towards gelatin. This indicates that glycosylation is not required for trafficking. Together these findings provide evidence that the major route of trafficking of Leishmania cysteine proteases to lysosomes is via the flagellar pocket and therefore differs significantly from cysteine protease trafficking in mammalian cells.
Assuntos
Cisteína Endopeptidases/genética , Cisteína Endopeptidases/metabolismo , Leishmania mexicana/enzimologia , Leishmania mexicana/genética , Animais , Sítios de Ligação , Cisteína , Cisteína Endopeptidases/química , Ativação Enzimática , Glicosilação , Lisossomos/enzimologia , Lisossomos/ultraestrutura , Microscopia Imunoeletrônica , Mutagênese Sítio-Dirigida , Proteínas Recombinantes/química , Proteínas Recombinantes/metabolismo , TransfecçãoRESUMO
Thanks to the phylogenetic systematics revolution, systematic parasitology is poised to make significant contributions in tropical medicine and public health, biodiversity science, and evolutionary biology. At the same time, the taxonomic impediment is acute within parasitology. Both systematists and non-systematists must be interested in working towards common goals and establishing collaborative efforts in order to re-vitalize and re-populate systematic parasitology.
Assuntos
Parasitos/classificação , Parasitologia/métodos , Filogenia , Animais , Ecossistema , Parasitos/genéticaRESUMO
Five species of Oochoristica, Oochoristica osheroffi, Oochoristica gracewileyae, Oochoristica whitentoni, and 2 new species described herein have strobilae longer than 200 mm, many secondary ovarian lobes, and testes extending anteriorly to midovarian level. A combination of 3 characters distinguishes the 5 species from each other. Oochoristica osheroffi has an average of 68 testes per proglottid, ovarian lobes wider than long, and osmoregulatory canals not forming an anastomosing plexus; O. gracewileyae has an average of 113 testes per proglottid, ovarian lobes longer than wide, and osmoregulatory canals not forming an anastomosing plexus; O. whitentoni has an average of 125 testes per proglottid, ovarian lobes wider than long, and osmoregulatory canals forming an anastomosing plexus; a new species in Ctenosaura similis from Costa Rica has an average of 62 testes per proglottid, ovarian lobes longer than wide, and osmoregulatory canals forming an anastomosing plexus; and a new species in Ctenosaura pectinata from Mexico has an average of 122 testes per progottid, ovarian lobes longer than wide, and osmoregulatory canals forming an anastomosing plexus. Oochoristica gracewileyae differs from the other 4 species by having genital pores 10-15% rather than 25-35% of proglottid length from the anterior end and by having convoluted rather than straight or sinuous transverse osmoregulatory canals. The new species from Mexico differs from the other 4 species and apparently from all described species of Oochoristica thus far by possessing darkly staining granules throughout the parenchyma of the scolex.
Assuntos
Cestoides/classificação , Infecções por Cestoides/veterinária , Lagartos/parasitologia , Animais , Cestoides/anatomia & histologia , Infecções por Cestoides/parasitologia , Costa Rica , MéxicoRESUMO
Molecular evidence is interpreted in the light of morphology to examine the validity of several species of Haematoloechus described as Mexican endemics. Internal transcribed spacers 1 and 2 and 28S ribosomal genes were sequenced for 11 isolates. Phylogenetic analysis of separate partitions and combined databases was conducted. Results were analyzed, in the light of morphological evidence. Haematoloechus macrorchis is proposed as a junior synonym of Haematoloechus longiplexus. Haematoloechus pulcher is a sibling species with Haematoloechus complexus in Lerma wetlands. In Mexico, Haematoloechus medioplexus is distributed along the east coast coinciding with the distribution of Rana berlandieri. The sister species of H. medioplexus is Haematoloechus coloradensis, sharing the distribution of the uterus as a synapomorphic character. Haematoloechus illimis is more closely related to H. medioplexus and H. coloradensis than to H. complexus. It can be distinguished by the distribution of the uterus, lobed ovary, and testes.
Assuntos
Ambystoma/parasitologia , Ranidae/parasitologia , Trematódeos/classificação , Infecções por Trematódeos/veterinária , Animais , Sequência de Bases , DNA de Helmintos/química , DNA Ribossômico/química , México , Dados de Sequência Molecular , Filogenia , RNA Ribossômico 28S/genética , Trematódeos/anatomia & histologia , Trematódeos/genética , Infecções por Trematódeos/parasitologiaRESUMO
A new species of Scyphophyllidium inhabits Mustelus mento near La Paloma, Uruguay. It resembles Scyphophyllidium giganteum from the Atlantic Ocean and specimens identified as S. giganteum from California by having anapolytic strobilae 155-258 mm long, 250-300 craspedote proglottids, scoleces 1.2-1.4 mm wide, necks 34-41 mm long, immature and mature proglottids wider than long, gravid proglottids wider than long to longer than wide, genital pores averaging 28% of proglottid length from the anterior end, relatively flat ovaries with digitiform lobes reaching the lateralmost extent of the testicular field, vitellaria in 2 fields converging toward the proglottid midline, straight and short cirrus sacs, and postvaginal vas deferens. The bothridia of the new species have accessory bothridial suckers that are smaller than those of California specimens; European specimens reportedly lack accessory bothridial suckers. The new species possesses a uterine duct that joins the uterus at the level of the genital atrium and ventral osmoregulatory ducts medial rather than lateral to the dorsal ducts, an arrangement described for Californian but not European specimens. It differs from both European and Californian specimens by having longer cirri, more testes per proglottid, prominent scales covering the neck, and vaginae and uterine ducts coiled immediately preovarially. Pithophorus, Marsupiobothrium, and Scyphophyllidium may form a clade.
Assuntos
Cestoides/classificação , Infecções por Cestoides/veterinária , Doenças dos Peixes/parasitologia , Tubarões/parasitologia , Animais , Cestoides/anatomia & histologia , Cestoides/ultraestrutura , Infecções por Cestoides/parasitologia , Microscopia Eletrônica de Varredura , Filogenia , UruguaiRESUMO
A new species of tetraphyllidean eucestode inhabiting Urobatis tumbesensis from inshore waters of southeastern Ecuador shares 3 synapomorphies with Rhinebothroides spp.: apical bothridial suckers poorly differentiated from the marginal loculi, internal seminal vesicles, and insertion of the vas deferens dorsally closer to the poral than the aporal end of the cirrus sac. The new species differs from Rhinebothroides spp. by lacking medial bothridial septa and loculi and having symmetrical ovarian arms, and possesses an apparent autapomorphic trait by having the vas deferens tapering to a narrow tube before entering the cirrus sac, extending posteriorly to the posterior end of the cirrus sac where it expands into an external seminal vesicle running ventral to the cirrus sac anteriorly to anterior to the vagina. In Rhinebothroides spp., the vas deferens is expanded into an external seminal vesicle near the insertion into the cirrus sac As the sister group of Rhinebothroides, we propose a new genus to accommodate the new species. Phylogenetic evaluation of phyllobothriids recently assigned to Anthocephalum shows that they represent a paraphyletic assemblage of species of varying degrees of relatedness to Rhinebothroides spp. and the new species. Uncovering the relationships of the new species and the various species assigned to Anthocephalum permitted reevaluation of character transformations used in previous phylogenetic analysis of Rhinebothroides. Transformation series for 3 characters, previously based on functional outgroup comparisons, changed and a new character, length of cirrus sac, was added. The new phylogenetic analysis differs from the previous hypothesis only in placing R. scorzai as the sister species of R. circularisi + R. venezuelae + R. moralarai rather than of R. freitasi + R. glandularis + R. mclennanae. The occurrence of the sister species of Rhinebothroides in a Pacific Ocean stingray adds additional support to the hypothesis of Pacific origins of South American freshwater stingrays.
Assuntos
Cestoides/classificação , Infecções por Cestoides/veterinária , Doenças dos Peixes/parasitologia , Rajidae/parasitologia , Animais , Cestoides/anatomia & histologia , Infecções por Cestoides/parasitologia , Equador , Oceano Pacífico , FilogeniaRESUMO
Echinocephalus janzeni n. sp. in the stingray, Himantura pacifica, is described from the eastern Pacific Ocean off the coasts of Costa Rica and southern Mexico. On the basis of the presence of 6 postanal caudal papillae, and modified annules anterior to the caudal alae in males, E. janzeni is most similar to Echinocephalus daileyi and Echinocephalus diazi. Specimens of E. janzeni are distinguished from those of E. daileyi by bilobed caudal alae and long cervical sacs that extend up to 65% of the length of the esophagus; E. janzeni is differentiated from E. diazi by the number of rows of cephalic spines (30-38 vs. 26-27), arrangement of the postanal caudal papillae, 3 rather than 2 preanal papillae, relative position and distance between the anus and vulva (395-460 microm vs. 70 microm), the digitiform female tail with a terminal cuticular fold, and the length of the female tail (450-480 microm vs. 270 microm). Cladistic analysis of the 10 Echinocephalus spp. resulted in a single most parsimonious tree (consistency index=0.893) and placed E. janzeni in a highly derived subclade where E. daileyi is the sister species of E. diazi + E. janzeni. Historical biogeographic analysis of hosts and parasites provides support for origins in the Pacific rather than the Atlantic for the potamotrygonid stingrays.