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1.
J Invertebr Pathol ; 154: 37-41, 2018 05.
Artigo em Inglês | MEDLINE | ID: mdl-29608918

RESUMO

Trypanosomatids are highly prevalent pathogens of Hymenoptera; however, most molecular methods used to detect them in Apis and Bombus spp. do not allow the identification of the infecting species, which then becomes expensive and time consuming. To overcome this drawback, we developed a multiplex PCR protocol to readily identify in a single reaction the main trypanosomatids present in these hymenopterans (Lotmaria passim, Crithidia mellificae and Crithidia bombi), which will facilitate the study of their epidemiology and transmission dynamics. A battery of primers, designed to simultaneously amplify fragments of the RNA polymerase II large subunit (RPB1) of L. passim, the glyceraldehyde-3-phosphate dehydrogenase (GAPDH) of C. mellificae and the DNA topoisomerase II (TOPII) of C. bombi, was tested for target specificity under single and mixed template conditions using DNA extracted from cell cultures (L. passim ATCC PRA403; C. mellificae ATCC 30254) and from a bumblebee specimen infected with C. bombi only (14_349). Once validated, the performance of the method was assessed using DNA extractions from seven Apis mellifera (Linnaeus, 1758) and five Bombus terrestris (Linnaeus, 1758) field samples infected with trypanosomatids whose identity had been previously determined by PCR-cloning and sequencing (P-C-S). The new method confirmed the results obtained by P-C-S: two of the honeybee samples were parasitized by L. passim, C. mellificae and C. bombi at the same time, whereas the other five were infected with L. passim only. The method confirmed the simultaneous presence of L. passim and C. mellificae in two B. terrestris, where these parasites had not previously been reported.


Assuntos
Abelhas/parasitologia , Reação em Cadeia da Polimerase Multiplex/métodos , Trypanosomatina/genética , Animais , Infecções por Euglenozoa/diagnóstico , Trypanosomatina/isolamento & purificação
2.
Mem Inst Oswaldo Cruz ; 113(1): 45-55, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29211107

RESUMO

BACKGROUND: Didelphis spp. are a South American marsupial species that are among the most ancient hosts for the Trypanosoma spp. OBJECTIVES: We characterise a new species (Trypanosoma janseni n. sp.) isolated from the spleen and liver tissues of Didelphis aurita in the Atlantic Rainforest of Rio de Janeiro, Brazil. METHODS: The parasites were isolated and a growth curve was performed in NNN and Schneider's media containing 10% foetal bovine serum. Parasite morphology was evaluated via light microscopy on Giemsa-stained culture smears, as well as scanning and transmission electron microscopy. Molecular taxonomy was based on a partial region (737-bp) of the small subunit (18S) ribosomal RNA gene and 708 bp of the nuclear marker, glycosomal glyceraldehyde-3-phosphate dehydrogenase (gGAPDH) genes. Maximum likelihood and Bayesian inference methods were used to perform a species coalescent analysis and to generate individual and concatenated gene trees. Divergence times among species that belong to the T. cruzi clade were also inferred. FINDINGS: In vitro growth curves demonstrated a very short log phase, achieving a maximum growth rate at day 3 followed by a sharp decline. Only epimastigote forms were observed under light and scanning microscopy. Transmission electron microscopy analysis showed structures typical to Trypanosoma spp., except one structure that presented as single-membraned, usually grouped in stacks of three or four. Phylogeography analyses confirmed the distinct species status of T. janseni n. sp. within the T. cruzi clade. Trypanosoma janseni n. sp. clusters with T. wauwau in a well-supported clade, which is exclusive and monophyletic. The separation of the South American T. wauwau + T. janseni coincides with the separation of the Southern Super Continent. CONCLUSIONS: This clade is a sister group of the trypanosomes found in Australian marsupials and its discovery sheds light on the initial diversification process based on what we currently know about the T. cruzi clade.


Assuntos
DNA de Protozoário/genética , Didelphis/parasitologia , RNA Ribossômico 18S/genética , Trypanosomatina/genética , Animais , Brasil , Filogeografia , Reação em Cadeia da Polimerase , Floresta Úmida , Trypanosoma cruzi , Trypanosomatina/classificação , Trypanosomatina/isolamento & purificação
3.
Parasitol Res ; 117(2): 623-628, 2018 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-29282526

RESUMO

Currently, light microscopic examination of cell morphology cannot discriminate Crithidia mellificae and Lotmaria passim with 100% certainty. Here, a minor groove-binding (MGB) probe-based multiplex real-time PCR assay was developed for the simultaneous and quantitative detection of C. mellificae and L. passim in honey bees. A conserved Hymenoptera 18S rRNA gene was built in as an internal control that allows accurate detection of PCR inhibition and failure of DNA extraction. The newly developed assay was also applied to field samples. Of 21 honey bee colonies (446 bees) sampled from six counties in both central and eastern Massachusetts, 3 colonies (14.29%) and 8 bees (1.79%) were infected with L. passim, and 1 colony (4.76%) and 1 bee (0.22%) with C. mellificae. Our data showed a low rate of trypanosomatid infection, and L. passim was more prevalent than C. mellificae in honey bee samples in Massachusetts.


Assuntos
Abelhas/parasitologia , Crithidia/isolamento & purificação , Reação em Cadeia da Polimerase Multiplex/métodos , Reação em Cadeia da Polimerase em Tempo Real/métodos , Trypanosomatina/isolamento & purificação , Animais , Crithidia/classificação , Crithidia/genética , Massachusetts , Trypanosomatina/classificação , Trypanosomatina/genética
4.
Am J Trop Med Hyg ; 97(5): 1482-1485, 2017 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-29140234

RESUMO

To better understand the ecology of Trypanosoma cruzi in the northeastern Peruvian Amazon, we evaluated the prevalence of T. cruzi and other trypanosomatids in four orders of wild mammals hunted and consumed by inhabitants of three remote indigenous communities in the Peruvian Amazon. Of 300 wild mammals sampled, 115 (38.3%) were infected with trypanosomatids and 15 (5.0%) with T. cruzi. The prevalence of T. cruzi within each species was as follows: large rodents (Cuniculus paca, 5.5%; Dasyprocta spp., 2.6%), edentates (Dasypus novemcinctus, 4.2%), and carnivores with higher prevalence (Nasua nasua, 18.8%). The high prevalence of T. cruzi and other trypanosomatids in frequently hunted wild mammals suggests a sizeable T. cruzi sylvatic reservoir in remote Amazonian locations.


Assuntos
Animais Selvagens/parasitologia , Doença de Chagas/veterinária , Mamíferos/parasitologia , Trypanosoma cruzi/isolamento & purificação , Trypanosomatina/isolamento & purificação , Animais , Tatus/parasitologia , Doença de Chagas/epidemiologia , Peru/epidemiologia , Prevalência , Procyonidae/parasitologia , Roedores/parasitologia , Trypanosoma cruzi/classificação , Trypanosomatina/classificação
5.
Arch Virol ; 162(12): 3827-3835, 2017 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-28939968

RESUMO

The great majority of kala-azar/visceral leishmaniasis (VL) cases, which are caused by Leishmania donovani (LD), are reported in Asia. We investigated whether leishmaniaviruses (LRVs) are present in LD isolates. These dsRNA viruses contribute to hyperpathogenicity, as observed in the case of other members of the genus Leishmania. However, LRVs could not be detected in 22 Indian LD isolates tested in the present study, while 70% of these original LD isolates harboured a virus that was not of LD but instead of Leptomonas seymouri (LS) origin. LS is another protozoon that parasitizes the sandfly vector of LD. Historically, LD clinical isolates from India often showed high incidence of LS coinfection. LS was detected in 20 out of the 22 (91%) above-mentioned LD isolates. Leptomonas seymouri narna-like virus 1 (Lepsey NLV1) was identified by whole-genome sequencing in an LD-LS coinfected sample, and its presence was confirmed by PCR and sequencing in 15 (75%) of the 20 LD-LS coinfected samples. The LS-negative LD samples were also virus negative by PCR. That the human host is exposed to an RNA virus in LS, another coinfecting parasite with LD, i.e., the "LD-LS-Lepsey NLV1 triple pathogen" phenomenon, unveils a new paradigm of research towards revisiting the mysteries of Indian leishmaniasis pathogenesis and management.


Assuntos
Leishmaniose Visceral/patologia , Leishmaniose Visceral/parasitologia , Vírus de RNA/classificação , Vírus de RNA/isolamento & purificação , Trypanosomatina/isolamento & purificação , Trypanosomatina/virologia , Genoma Viral , Humanos , Índia , Leishmania donovani/isolamento & purificação , Leishmania donovani/virologia , Reação em Cadeia da Polimerase , Vírus de RNA/genética , Análise de Sequência de DNA
6.
Parasit Vectors ; 10(1): 287, 2017 Jun 08.
Artigo em Inglês | MEDLINE | ID: mdl-28595622

RESUMO

Trypanosomatids are protozoan parasites of the class Kinetoplastida predominately restricted to invertebrate hosts (i.e. possess a monoxenous life-cycle). However, several genera are pathogenic to humans, animals and plants, and have an invertebrate vector that facilitates their transmission (i.e. possess a dixenous life-cycle). Phytomonas is one dixenous genus that includes several plant pathogens transmitted by phytophagous insects. Trypanosoma and Leishmania are dixenous genera that infect vertebrates, including humans, and are transmitted by hematophagous invertebrates. Traditionally, monoxenous trypanosomatids such as Leptomonas were distinguished from morphologically similar dixenous species based on their restriction to an invertebrate host. Nonetheless, this criterion is somewhat flawed as exemplified by Leptomonas seymouri which reportedly infects vertebrates opportunistically. Similarly, Novymonas and Zelonia are presumably monoxenous genera yet sit comfortably in the dixenous clade occupied by Leishmania. The isolation of Leishmania macropodum from a biting midge (Forcipomyia spp.) rather than a phlebotomine sand fly calls into question the exclusivity of the Leishmania-sand fly relationship, and its suitability for defining the Leishmania genus. It is now accepted that classic genus-defining characteristics based on parasite morphology and host range are insufficient to form the sole basis of trypanosomatid taxonomy as this has led to several instances of paraphyly. While improvements have been made, resolution of evolutionary relationships within the Trypanosomatidae is confounded by our incomplete knowledge of its true diversity. The known trypanosomatids probably represent a fraction of those that exist and isolation of new species will help resolve relationships in this group with greater accuracy. This review incites a dialogue on how our understanding of the relationships between certain trypanosomatids has shifted, and discusses new knowledge that informs the present taxonomy of these important parasites.


Assuntos
Filogenia , Trypanosomatina/classificação , Animais , Plantas , Trypanosomatina/isolamento & purificação
7.
J Invertebr Pathol ; 134: 1-5, 2016 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-26721451

RESUMO

The recently described trypanosome Lotmaria passim is currently considered the most predominant trypanosomatid in honey bees worldwide and could be a factor in honey bee declines. For a specific and quick detection of this pathogen, we developed primers based on the SSU rRNA and gGAPDH genes for the detection of L. passim in Chilean honey beehives. PCR products amplified and sequenced for these primers shared 99-100% identity with other sequences of L. passim. The designed primers were specific and we were able to detect a high prevalence (40-90%) of L. passim in bee hives distributed throughout Chile. Our described PCR-based method offers a feasible and specific detection of L. passim in any honey bee samples.


Assuntos
Abelhas/parasitologia , Trypanosomatina/genética , Animais , Chile , Primers do DNA , DNA de Protozoário/química , Filogenia , Reação em Cadeia da Polimerase/métodos , Trypanosomatina/isolamento & purificação
8.
J Eukaryot Microbiol ; 63(2): 198-209, 2016.
Artigo em Inglês | MEDLINE | ID: mdl-26352484

RESUMO

In this study, we surveyed six species of cockroaches, two synanthropic (i.e. ecologically associated with humans) and four wild, for intestinal trypanosomatid infections. Only the wild cockroach species were found to be infected, with flagellates of the genus Herpetomonas. Two distinct genotypes were documented, one of which was described as a new species, Herpetomonas tarakana sp. n. We also propose a revision of the genus Herpetomonas and creation of a new subfamily, Phytomonadinae, to include Herpetomonas, Phytomonas, and a newly described genus Lafontella n. gen. (type species Lafontella mariadeanei comb. n.), which can be distinguished from others by morphological and molecular traits.


Assuntos
Baratas/parasitologia , Trypanosomatina/classificação , Animais , Biodiversidade , República Tcheca , DNA de Protozoário/genética , Genótipo , Microscopia Eletrônica de Transmissão , Filogenia , Reação em Cadeia da Polimerase/métodos , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Eslováquia , Trypanosomatina/genética , Trypanosomatina/isolamento & purificação , Trypanosomatina/ultraestrutura
9.
Rev Soc Bras Med Trop ; 48(6): 776-9, 2015.
Artigo em Inglês | MEDLINE | ID: mdl-26676507

RESUMO

INTRODUCTION: This study describes the occurrence of trypanosomatids in phlebotomines in Brasília, Brazil. METHODS: Two hundred and ten females of 13 sand fly species were analyzed by polymerase chain reaction (PCR) using different molecular markers (D7 24Sα rRNA, kDNA, and ITS1) and sequencing. RESULTS: PCR revealed trypanosomatid-positive samples from Nyssomyia whitmani and Evandromyia evandroi, which were negative by kDNA and ITS1 Leishmania-specific PCRs. DNA sequence analysis of D7 24Sα rRNA amplicons indicated the occurrence of Blastocrithidia sp. and Trypanosoma sp. in Nyssomyia whitmani and Evandromyia evandroi, respectively. CONCLUSIONS: Two trypanosomatid species other than Leishmania sp. were found to circulate in sand flies in Central Brazil.


Assuntos
DNA de Cinetoplasto/genética , DNA de Protozoário/genética , Insetos Vetores/parasitologia , Psychodidae/parasitologia , Trypanosomatina/genética , Animais , Brasil , Feminino , Insetos Vetores/classificação , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Psychodidae/classificação , Trypanosomatina/classificação , Trypanosomatina/isolamento & purificação
10.
Parasitology ; 142(14): 1657-62, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26492813

RESUMO

Protozoan parasites Leishmania donovani (family: Trypanosomatidae) cause fatal visceral leishmaniasis (VL) and the infection relapses in apparently cured population as post kala-azar dermal leishmaniasis (PKDL) in the Indian subcontinent. In recent years co-infection of another Trypanosomatid parasite Leptomonas with L. donovani during VL/PKDL in this region has become prominent. The observation of clinically lesser-known insect parasite, Leptomonas in leishmaniasis is intriguing to researchers. The presence of Leishmania look alike Leptomonas in the cultures of clinical isolates of Leishmania has been worrisome to those, who prefer to work with pure Leishmania cultures for drug and vaccine development or immune response studies. The exact implications of such a co-habitation, which might lead to a delay in the diagnostics of VL and elevate mortality, need a thorough investigation. Also whether Leptomonas is involved in leishmaniasis manifestation needs to be ascertained. Thus we are currently witnessing a new paradigm of a parasitic co-infection in VL/PKDL cases in India and this review outlines various opportunities for further research in understanding such emerging co-infection.


Assuntos
Doenças Transmissíveis Emergentes/complicações , Infecções por Euglenozoa/complicações , Leishmania donovani/isolamento & purificação , Leishmaniose Visceral/complicações , Trypanosomatina/isolamento & purificação , Coinfecção , Doenças Transmissíveis Emergentes/epidemiologia , Infecções por Euglenozoa/epidemiologia , Índia/epidemiologia , Leishmaniose Visceral/epidemiologia
11.
J Med Entomol ; 52(3): 461-8, 2015 May.
Artigo em Inglês | MEDLINE | ID: mdl-26334822

RESUMO

Multiple mosquito-borne parasites cocirculate in nature and potentially interact. To understand the community of parasites cocirculating with West Nile virus (WNV), we screened the bloodmeal content of Culex pipiens L. mosquitoes for three common types of hemoparasites. Blood-fed Cx. pipiens were collected from a WNV-epidemic area in suburban Chicago, IL, from May to September 2005 through 2010. DNA was extracted from dissected abdomens and subject to PCR and direct sequencing to identify the vertebrate host. RNA was extracted from the head or thorax and screened for WNV using quantitative reverse transcriptase PCR. Seventy-nine engorged females with avian host origin were screened using PCR and amplicon sequencing for filarioid nematodes, Haemosporida, and trypanosomatids. Filarioid nematodes were identified in 3.8% of the blooded abdomens, Plasmodium sp. in 8.9%, Haemoproteus in 31.6%, and Trypanosoma sp. in 6.3%. The sequences from these hemoparasite lineages were highly similar to sequences from birds in prior studies in suburban Chicago. Overall, 50.6% of blood-fed Culex pipiens contained hemoparasite DNA in their abdomen, presumably from current or prior bloodmeals. Additionally, we detected hemoparasite DNA in the blooded abdomen of three of 10 Cx. pipiens infected with WNV.


Assuntos
Culex/parasitologia , Filarioidea/isolamento & purificação , Haemosporida/isolamento & purificação , Trypanosomatina/isolamento & purificação , Animais , Columbidae/parasitologia , DNA/isolamento & purificação , DNA de Helmintos/isolamento & purificação , DNA de Protozoário/isolamento & purificação , Filarioidea/classificação , Filarioidea/genética , Haemosporida/classificação , Haemosporida/genética , Illinois , Dados de Sequência Molecular , Filogenia , Análise de Sequência de DNA/veterinária , Aves Canoras/parasitologia , Trypanosomatina/classificação , Trypanosomatina/genética , Febre do Nilo Ocidental/epidemiologia , Febre do Nilo Ocidental/etiologia , Febre do Nilo Ocidental/veterinária
12.
Med Vet Entomol ; 29(4): 444-7, 2015 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-26211924

RESUMO

Trypanosomes are known as widespread blood parasites of birds; however, knowledge of their prevalences in vectors and their overall biodiversity is rather limited. To assess the prevalences in potential vectors, we have microscopically examined ornithophilic bloodsucking Diptera (Culicidae, Simuliidae and Hippoboscidae) for the presence of trypanosomatids in their guts. In total, 3270 specimens were dissected, namely Culex pipiens Linnaeus, 1758 (n = 898), C. modestus Ficalbi, 1890 (136), Simulium vernum (Macquart, 1838) (1455), S. angustipes Edwards, 1915 (221) and Ornithomyia avicularia (Linnaeus, 1758) (560). All insect species were found to be infected with trypanosomatids, and the prevalence ranged from 4 to 8% but reached 60% in S. vernum. Blackflies and hippoboscids exclusively harboured trypanosomes (both T. cf. avium s.s. Danilewsky, 1885; T. corvi/culicavium group in hippoboscids). Mosquitoes were infected with T. culicavium Votypka, 2012 and T. avium s. l. but also with monoxenous parasites, namely Crithidia brevicula Frolov and Malysheva, 1989, and Paratrypanosoma confusum Votypka and Lukes, 2013. Only 4% of the isolated parasite strains were monoxenous whereas the majority were avian trypanosomes, confirming the vectorial status of the studied insects.


Assuntos
Dípteros/parasitologia , Insetos Vetores/parasitologia , Trypanosomatina/isolamento & purificação , Animais , Culicidae/parasitologia , República Tcheca , Aves Predatórias/parasitologia , Simuliidae/parasitologia
13.
Parasitol Int ; 64(4): 1-5, 2015 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-25582929

RESUMO

Leishmania and Leptomonas are protozoan parasites of the family Trypanosomatidae. Leishmania donovani causes the fatal visceral leishmaniasis (VL; kala-azar) in mammals and is transmitted by sand fly vector. Certain VL-cured human populations in India and Sudan develop post kala-azar dermal leishmaniasis (PKDL) due to the same parasite. Although Leptomonas is parasitic mainly in insects, several recent reports on the clinical isolates of L. donovani from VL and PKDL patients in India confirm co-infection of Leptomonas seymouri, probably due to immune suppression in those individuals. Detection of L. seymouri in the in vitro cultures of L. donovani from clinical origin is difficult due to many similarities between L. seymouri and L. donovani. We describe here ways to detect L. seymouri and L. donovani in co-culture. In addition, based on our observation regarding the growth of L. seymouri in different culture conditions, we report here a novel procedure, which can selectively eliminate L. seymouri from the in vitro co-culture with L. donovani. This would be beneficial to researchers who prefer to deal with pure populations of Leishmania parasites for various downstream immunological and genetic studies.


Assuntos
Leishmania donovani/crescimento & desenvolvimento , Leishmania donovani/isolamento & purificação , Parasitologia/métodos , Trypanosomatina/crescimento & desenvolvimento , Trypanosomatina/isolamento & purificação , Técnicas de Cocultura , Meios de Cultura , Humanos , Leishmania donovani/genética , Reação em Cadeia da Polimerase , Polimorfismo de Fragmento de Restrição , Temperatura , Trypanosomatina/genética
14.
Protist ; 165(6): 825-38, 2014 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-25460233

RESUMO

Compared to their relatives, the diversity of endosymbiont-containing Trypanosomatidae remains under-investigated, with only two new species described in the past 25 years, bringing the total to six. The possible reasons for such a poor representation of this group are either their overall scarcity or susceptibility of their symbionts to antibiotics that are traditionally used for cultivation of flagellates. In this work we describe the isolation, cultivation, as well as morphological and molecular characterization of a novel endosymbiont-harboring trypanosomatid species, Kentomonas sorsogonicus sp. n. The newly erected genus Kentomonas gen. n. shares many common features with the genera Angomonas and Strigomonas, such as the presence of an extensive system of peripheral mitochondrial branches distorting the corset of subpellicular microtubules, large and loosely packed kinetoplast, and a rudimentary paraflagellar rod. Here we also propose to unite all endosymbiont-bearing trypanosomatids into the new subfamily Strigomonadinae subfam. n.


Assuntos
Filogenia , Simbiose/genética , Trypanosomatina/classificação , Animais , Teorema de Bayes , Clonagem Molecular , DNA Bacteriano/isolamento & purificação , DNA Viral/isolamento & purificação , Funções Verossimilhança , Modelos Genéticos , RNA de Protozoário/genética , RNA Ribossômico 18S/genética , Sarcofagídeos/parasitologia , Análise de Sequência de DNA , Especificidade da Espécie , Trypanosomatina/isolamento & purificação , Trypanosomatina/microbiologia , Trypanosomatina/virologia
15.
BMC Res Notes ; 7: 649, 2014 Sep 15.
Artigo em Inglês | MEDLINE | ID: mdl-25223634

RESUMO

BACKGROUND: Here we present a holistic screening of collapsing colonies from three professional apiaries in Spain. Colonies with typical honey bee depopulation symptoms were selected for multiple possible factors to reveal the causes of collapse. RESULTS: Omnipresent were Nosema ceranae and Lake Sinai Virus. Moderate prevalences were found for Black Queen Cell Virus and trypanosomatids, whereas Deformed Wing Virus, Aphid Lethal Paralysis Virus strain Brookings and neogregarines were rarely detected. Other viruses, Nosema apis, Acarapis woodi and Varroa destructor were not detected. Palinologic study of pollen demonstrated that all colonies were foraging on wild vegetation. Consequently, the pesticide residue analysis was negative for neonicotinoids. The genetic analysis of trypanosomatids GAPDH gene, showed that there is a large genetic distance between Crithidia mellificae ATCC30254, an authenticated cell strain since 1974, and the rest of the presumed C. mellificae sequences obtained in our study or published. This means that the latter group corresponds to a highly differentiated taxon that should be renamed accordingly. CONCLUSION: The results of this study demonstrate that the drivers of colony collapse may differ between geographic regions with different environmental conditions, or with different beekeeping and agricultural practices. The role of other pathogens in colony collapse has to bee studied in future, especially trypanosomatids and neogregarines. Beside their pathological effect on honey bees, classification and taxonomy of these protozoan parasites should also be clarified.


Assuntos
Criação de Abelhas/métodos , Abelhas , Colapso da Colônia , Vírus de Insetos/patogenicidade , Nosema/patogenicidade , Trypanosomatina/patogenicidade , Animais , Abelhas/microbiologia , Abelhas/parasitologia , Abelhas/virologia , Colapso da Colônia/microbiologia , Colapso da Colônia/parasitologia , Colapso da Colônia/virologia , Ecossistema , Comportamento Alimentar , Interações Hospedeiro-Parasita , Interações Hospedeiro-Patógeno , Vírus de Insetos/genética , Vírus de Insetos/isolamento & purificação , Nosema/genética , Nosema/isolamento & purificação , Filogenia , Pólen , Dinâmica Populacional , Ribotipagem , Espanha , Trypanosomatina/genética , Trypanosomatina/isolamento & purificação
16.
Parasit Vectors ; 7: 203, 2014 Apr 28.
Artigo em Inglês | MEDLINE | ID: mdl-24774858

RESUMO

BACKGROUND: Flagellates of the family Trypanosomatidae are obligate endoparasites, which can be found in various hosts. Several genera infect insects and occur as monoxenous parasites especially in representatives of Diptera and Hemiptera. These trypanosomatid flagellates probably share the worldwide distribution of their hosts, which are often infested by large numbers of endoparasites. Traditionally, their taxonomy was based on morphology, host origin, and life cycle. Here we report the characterization of a trypanosomatid infection detected in a protozoan, a ciliate collected from a polluted freshwater pond in a suburb of New Delhi (India). METHODS: Live observations and morphological studies applying light, fluorescence and transmission electron microscopy were conducted. Molecular analyses of host and parasite were performed and used for phylogenetic reconstructions and species (host) or genus level (parasite) identification. RESULTS: Although the morphological characteristics were not revealing, a high similarity of the trypanosomatids 18S rRNA gene sequence to Herpetomonas ztiplika and Herpetomonas trimorpha (Kinetoplastida, Trypanosomatidae), both parasites of biting midges (Culicoides kibunensis and Culicoides truncorum, respectively) allowed the assignment to this genus. The majority of the host population displayed a heavy infection that significantly affected the shape of the host macronucleus, which was the main site of parasite localization. In addition, the growth rate of host cultures, identified as Euplotes encysticus according to cell morphology and 18S rRNA gene sequence, was severely impacted by the infection. CONCLUSIONS: The host-parasite system described here represents a recent example of free-living protists acting as environmental reservoirs for parasitic eukaryotic microorganisms.


Assuntos
Euplotes/parasitologia , Macronúcleo , Trypanosomatina/isolamento & purificação , Animais , Reservatórios de Doenças , Interações Hospedeiro-Parasita , Filogenia , RNA Ribossômico 18S/genética , Trypanosomatina/classificação , Trypanosomatina/genética
17.
Protist ; 164(6): 763-81, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24113136

RESUMO

To further investigate the diversity of Trypanosomatidae we have examined the species present within the flea (Siphonaptera) population in the Czech Republic. Out of 1549 fleas, 239 were found to be infected by trypanosomatids. Axenic cultures were established from 90 infected specimens and 29 of them were further characterized. Molecular phylogenetic analysis of the SL RNA, gGAPDH, and SSU rRNA genes revealed a striking diversity within this group and analyzed isolates were classified into 16 Typing units (TUs) of which 15 typified new species. In addition to one Trypanosoma species, two TUs grouped within the sub-family Leishmaniinae, two clustered together with Herpetomonas, wheras 11 TUs formed a novel clade branching off between Trypanosoma spp. and remaining trypanosomatids. We propose to recognize this clade as a new genus Blechomonas and a new subfamily Blechomonadinae, and provide molecular and morphological description of 11 TUs representing this genus. Our finding of such an ancient host-specific group sheds new light at the origin of Trypanosomatidae and the roots of dixenous parasitism. The strict host restriction of Blechomonas to Siphonaptera with adult fleas' dependence on blood meal may reflect passing of parasites from larvae through pupae to adults and implies potential transmission to the warm-blooded vertebrates.


Assuntos
Biodiversidade , Sifonápteros/parasitologia , Trypanosomatina/classificação , Trypanosomatina/isolamento & purificação , Animais , República Tcheca , Dados de Sequência Molecular , Filogenia , Roedores , Sifonápteros/classificação , Sifonápteros/genética , Trypanosomatina/genética , Trypanosomatina/fisiologia
18.
Curr Biol ; 23(18): 1787-93, 2013 Sep 23.
Artigo em Inglês | MEDLINE | ID: mdl-24012313

RESUMO

The kinetoplastids are a widespread and important group of single-celled eukaryotes, many of which are devastating parasites of animals, including humans. We have discovered a new insect trypanosomatid in the gut of Culex pipiens mosquitoes. Glyceraldehyde-3-phosphate dehydrogenase- and SSU rRNA-based phylogenetic analyses show this parasite to constitute a distinct branch between the free-living Bodo saltans and the obligatory parasitic clades represented by the genus Trypanosoma and other trypanosomatids. From draft genome sequence data, we identified 114 protein genes shared among the new flagellate, 15 trypanosomatid species, B. saltans, and the heterolobosean Naegleria gruberi, as well as 129 protein genes shared with the basal kinetoplastid Perkinsela sp. Individual protein phylogenies together with analyses of concatenated alignments show that the new species, here named Paratrypanosoma confusum n. gen., n. sp., branches with very high support at the base of the family Trypanosomatidae. P. confusum thus represents a long-sought-after missing link between the ancestral free-living bodonids and the derived parasitic trypanosomatids. Further analysis of the P. confusum genome should provide insight into the emergence of parasitism in the medically important trypanosomatids.


Assuntos
Filogenia , Trypanosomatina/classificação , Sequência de Aminoácidos , Animais , Culex/parasitologia , Feminino , Genoma , Dados de Sequência Molecular , RNA Ribossômico/química , Alinhamento de Sequência , Análise de Sequência de Proteína , Análise de Sequência de RNA , Trypanosomatina/genética , Trypanosomatina/isolamento & purificação , Trypanosomatina/ultraestrutura
19.
PLoS One ; 8(2): e55738, 2013.
Artigo em Inglês | MEDLINE | ID: mdl-23418454

RESUMO

BACKGROUND: Known as 'neglected disease' because relatively little effort has been applied to finding cures, leishmaniasis kills more than 150,000 people every year and debilitates millions more. Visceral leishmaniasis (VL), also called Kala Azar (KA) or black fever in India, claims around 20,000 lives every year. Whole genome analysis presents an excellent means to identify new targets for drugs, vaccine and diagnostics development, and also provide an avenue into the biological basis of parasite virulence in the L. donovani complex prevalent in India. METHODOLOGY/PRINCIPAL FINDINGS: In our presently described study, the next generation SOLiD™ platform was successfully utilized for the first time to carry out whole genome sequencing of L. donovani clinical isolates from India. We report the exceptional occurrence of insect trypanosomatids in clinical cases of visceral leishmaniasis (Kala Azar) patients in India. We confirm with whole genome sequencing analysis data that isolates which were sequenced from Kala Azar (visceral leishmaniasis) cases were genetically related to Leptomonas. The co-infection in splenic aspirate of these patients with a species of Leptomonas and how likely is it that the infection might be pathogenic, are key questions which need to be investigated. We discuss our results in the context of some important probable hypothesis in this article. CONCLUSIONS/SIGNIFICANCE: Our intriguing results of unusual cases of Kala Azar found to be most similar to Leptomonas species put forth important clinical implications for the treatment of Kala Azar in India. Leptomonas have been shown to be highly susceptible to several standard leishmaniacides in vitro. There is very little divergence among these two species viz. Leishmania sp. and L. seymouri, in terms of genomic sequence and organization. A more extensive perception of the phenomenon of co-infection needs to be addressed from molecular pathogenesis and eco-epidemiological standpoint.


Assuntos
Infecções por Euglenozoa/parasitologia , Leishmania donovani/genética , Leishmaniose Visceral/parasitologia , Trypanosomatina/genética , Coinfecção , Genoma de Protozoário , Humanos , Índia , Leishmania donovani/isolamento & purificação , Trypanosomatina/isolamento & purificação
20.
Protist ; 164(1): 129-52, 2013 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22938923

RESUMO

In order to review the taxonomy of the genus Herpetomonas through phylogenetic and morphological analyses we barcoded 527 insect trypanosomatids by sequencing the V7V8 region of the small subunit ribosomal RNA (SSU rRNA) gene. Fifty two flagellates, 90% of them from Diptera, revealed to be related to known species of Herpetomonas. Sequences of entire glycosomal glyceraldehyde phosphate dehydrogenase (gGAPDH) and SSU rRNA genes were employed for phylogenetic inferences including representatives of all genera of Trypanosomatidae. In the resulting phylogenetic trees, the selected flagellates clustered into a monophyletic assemblage that we are considering as the redefined genus Herpetomonas. Internal transcribed spacer 1 (ITS1) rDNA sequences and putative secondary structures of this region were compared for evaluation of inter- and intraspecific variability. The flagellates were classified in six already known species and five new species. In addition, two Leptomonas spp. were moved to Herpetomonas, now comprising 13 valid species, while four species were excluded from the genus. Light and electron microscopy revealed the extreme polymorphism of Herpetomonas, hindering genus and species identification by morphological characteristics. Our findings also showed that some species of Herpetomonas are generalist parasites of flies and appear to be as cosmopolitan as their hosts.


Assuntos
Dípteros/parasitologia , Trypanosomatina/classificação , Trypanosomatina/genética , Animais , Análise por Conglomerados , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , DNA Espaçador Ribossômico/química , DNA Espaçador Ribossômico/genética , Genes de RNAr , Gliceraldeído-3-Fosfato Desidrogenases/genética , Microscopia , Modelos Moleculares , Dados de Sequência Molecular , Conformação de Ácido Nucleico , Filogenia , RNA de Protozoário/genética , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Trypanosomatina/citologia , Trypanosomatina/isolamento & purificação
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