RESUMO
PURPOSE: Brazilian anurans are considered the most diverse and species rich around the world. Although in recent years there has been a strong focus on research related to this group of animals, their parasites have not received the same attention. Thus, this study aimed to provide morphological and molecular data on haemogregarines biodiversity infecting Brazilian anurans. METHODS: During 2020, 116 anurans were collected from four Brazilian States and their blood and fragment of organs were screened for haemogregarine parasites. RESULTS: From the total, seven (6.03%) animals were found infected with species of Hepatozoon and Dactylosoma. Based on the morphological and molecular analysis, four anurans were found infected with Hepatozoon latrensis. The phylogenetic analysis has shown the isolates from this study grouping with the Brazilian anuran Hepatozoon clade, also with gene similarity ranging from 99.70 to 100% to H. latrensis isolates available on GenBank. Furthermore, three specimens (Trachycephalus typhonius, Leptodactylus latrans, and Rhinella diptycha) were infected with the same species of Dactylosoma (100% genetic similarity), with a genetic similarity of 98.56% to Dactylosoma piperis the only other species described in Brazil. In support of the molecular data, different morphological characters were observed in the blood smears as compared to D. piperis, suggesting that the species of Dactylosoma from the present study infecting three different species of Brazilian anurans is an undescribed species. CONCLUSION: Thus, this study increases the knowledge of Brazilian anuran blood parasites and demonstrates the importance of using integrative approaches for the diagnosis of haemoparasites.
Assuntos
Apicomplexa , Eucoccidiida , Parasitos , Animais , Brasil , Filogenia , Apicomplexa/genética , Anuros/parasitologiaRESUMO
Although Brazil is a hotspot for snake species, there is a lack of information on the biodiversity of haemoparasites infecting these hosts. Thus, the present study aimed to bring new insights on the diversity of species of Hepatozoon (Apicomplexa: Adeleorina: Hepatozoidae) infecting Brazilian snakes from the Midwest and Southeast regions. The snakes were captured from 2018 to 2020 from the states of Mato Grosso, Mato Grosso do Sul, Goiás, and São Paulo. Three to five blood smears were made and the remaining blood sample was stored for further molecular analysis. Moreover, histopathological slides of the organs were stained with haematoxylin-eosin. Regarding molecular diagnosis, PCR was performed targeting different regions of the 18S rRNA gene of apicomplexan parasites. From the 13 free-living snakes screened, ten (76.92%) were found infected with Hepatozoon spp. Based on morphological and morphometric tools, five different morphotypes of species of Hepatozoon gamonts were detected. Molecular data and phylogenetic analysis support the morphological data, identifying five species of Hepatozoon from snakes, of which three species belong to previously described species, Hepatozoon cevapii, Hepatozoon cuestensis, and Hepatozoon quagliattus, with a genetic similarity of 100% (based on the 18S rRNA genetic marker). The present study identifies and describes two new species of Hepatozoon, Hepatozoon annulatum sp. nov. infecting the snake Leptodeira annulata and Hepatozoon trigeminum sp. nov. infecting the snake Oxyrhopus trigeminus. Thus, based on morphological and molecular data the present study provides new insights on haemogregarine diversity infecting Brazilian snakes from the Midwest and Southeast regions.
Assuntos
Apicomplexa , Coccidiose , Eucoccidiida , Parasitos , Animais , Apicomplexa/genética , Brasil/epidemiologia , Coccidiose/parasitologia , Coccidiose/veterinária , Eucoccidiida/genética , Parasitos/genética , Filogenia , RNA Ribossômico 18S/genética , Serpentes/parasitologiaRESUMO
The Amphibia are considered the most threatened vertebrate class globally, yet in Brazil they are also one of the more diverse and species rich groups. Although, in recent years there has been strong focus on amphibian related research, their parasites have not received the same attention. In Brazil, only a single species of Hepatozoon, namely H. leptodactyli (Lesage, 1908) Pessoa, 1970, has been described from anuran hosts. The present study aimed to describe three new species of Hepatozoon parasitising Leptodactylus labyrinthicus and Leptodactylus latrans from Mato Grosso State, Brazil. From 66 anurans screened for haemogregarines, four belonging to the Leptodactylidae were found positive for species of Hepatozoon. Based on the morphological analysis of peripheral blood gamonts and spleen and liver tissue meronts, three different morphotypes of Hepatozoon spp. were identified. Morphotype 1 (M1) and morphotype 2 (M2) in L. labyrinthicus and morphotype 3 (M3) in L. latrans. Molecular data based on partial 18S rDNA sequences revealed an interspecific divergence, between the species ranging from 0.43% to 1.16%. Phylogenetic analysis recovered isolates from the present study monophyletic with other isolates from Brazilian reptile and anuran hosts, sister to a clade comprising species isolated from African, North American and European reptile and anuran host species. Thus, using morphological and molecular analysis three new species infecting Brazilian Leptodactylidae anurans were identified and described. This study increases the knowledge of Brazilian anurans blood parasites and demonstrates the importance of using integrative approaches for diagnosis of hemoparasites.
RESUMO
PURPOSE: Species of Hepatozoon Miller, 1908 (Hepatozoidae) are blood protozoans with a cosmopolitan distribution and are reported to parasitize a range of vertebrate hosts including mammals, birds, reptiles, and amphibians. The present study aimed to describe a new species of Hepatozoon (Apicomplexa: Adeleorina: Hepatozoidae) found infecting the sleep snake Dipsas mikanii (Schlegel, 1837) (Squamata: Colubridae: Dipsadinae). METHODS: The snake was collected in 2017 at the municipality of Britânia, Goiás State, Brazil. Blood smears were made in order to find blood gametocytes and PCR was performed targeting the 18S rRNA gene. RESULTS: Microscopy screening of blood smears revealed the presence of intraerythrocytic gamont stages of Hepatozoon sp. in the peripheral blood with a parasitemia of 0.25%. Furthermore, meronts and monozoic cysts were observed in histological sections of the liver from the infected individual. The interspecific divergence of 18S rRNA sequences fragments isolated from D. mikanii had differences (2.39-11.3%) as compared to other sequences of species of Hepatozoon from snakes. CONCLUSIONS: Based on morphological and molecular data, a new species of Hepatozoon infecting D. mikanii from Brazil is described.
Assuntos
Apicomplexa , Coccidiose , Colubridae , Eucoccidiida , Lagartos , Animais , Brasil , Coccidiose/epidemiologia , Coccidiose/veterinária , Eucoccidiida/genética , Filogenia , SonoRESUMO
The Dactylosomatidae Jakowska and Negrelli, 1955 are one of four families belonging to adeleorinid coccidia and comprise the genera Babesiosoma Jakowska and Nigrelli, 1956 and Dactylosoma Labbé, 1894. These blood protozoa occur in peripheral blood of lower vertebrates, and are commonly reported parasitising amphibians. The present study describes Dactylosoma piperis n. sp. from the pepper frog Leptodactylus labyrinthicus (Spix, 1824) (Anura: Leptodactylidae), collected in 2018 at the municipality of Araguaiana, Mato Grosso State, Brazil, based on morphology of intra-erythrocytic trophozoite, primary and secondary merogonic stages and a molecular analysis (partial 18S rDNA). Dactylosoma piperis n. sp. forms a well-supported clade with other Dactylosomatidae. This is the first molecular characterization of a species of Dactylosoma from a Brazilian anuran.
TITLE: Une nouvelle espèce, Dactylosoma piperis n. sp. (Apicomplexa, Dactylosomatidae), parasite de la grenouille Leptodactylus labyrinthicus (Anura, Leptodactylidae) de l'état du Mato Grosso, Brésil. ABSTRACT: Les Dactylosomatidae Jakowska et Negrelli, 1955 sont l'une des quatre familles appartenant aux coccidies Adeleorina et comprennent les genres Babesiosoma Jakowska et Nigrelli, 1956 et Dactylosoma Labbé, 1894. Ces protozoaires sanguins se trouvent dans le sang périphérique des vertébrés inférieurs et sont fréquemment signalés comme parasitant des amphibiens. Ce travail décrit Dactylosoma piperis n. sp. de la grenouille Leptodactylus labyrinthicus (Spix, 1824) (Anura : Leptodactylidae), collectée en 2018 dans la municipalité d'Araguaiana, État du Mato Grosso, Brésil, d'après la morphologie du trophozoïte intra-érythrocytaire, des stades mérogoniques primaires et secondaires et une analyse moléculaire (ADNr 18S partiel). Dactylosoma piperis n. sp. forme un clade bien soutenu avec d'autres Dactylosomatidae. Il s'agit de la première caractérisation moléculaire d'une espèce de Dactylosoma à partir d'un anoure brésilien.
Assuntos
Anuros , Coccídios , Animais , Anuros/parasitologia , Brasil , Coccídios/classificação , Coccídios/citologia , Coccídios/genética , Filogenia , RNA Ribossômico 18S/genética , Especificidade da EspécieRESUMO
Hepatozoon pyramidumi sp. n. is described from the blood of the Egyptian saw-scaled viper, Echis pyramidum, captured from Saudi Arabia. Five out of ten viper specimens examined (50%) were found infected with Hepatozoon pyramidumi sp. n. with parasitaemia level ranged from 20-30%. The infection was restricted only to the erythrocytes. Two morphologically different forms of intraerythrocytic stages were observed; small and mature gamonts. The small ganomt with average size of 10.7 × 3.5 μm. Mature gamont was sausage-shaped with recurved poles measuring 16.3 × 4.2 μm in average size. Infected erythrocytes were hypertrophied; their nuclei were deformed and sometimes displaced from their central position in the normal uninfected cell. Merogonic stages were observed in the lung endothelial cell and the liver parenchyma cells. Mature meront was 17.8 × 13.6 µm and contained banana-shaped merozoites with average size of ~15 × 2 µm. Phylogenetic analysis based on the SSU rDNA sequence clustered Hepatozoon pyramidumi sp. n with previously sequenced Hepatozoon spp., most of them infected reptilian hosts without geographic consideration. The morphological and molecular comparison with closely related species proved the taxonomic uniqueness and novelty of the present form.(AU)
Hepatozoon pyramidumi sp. n. é descrito a partir do sangue da víbora em escamas e quilhas serrilhadas, Echis pyramidum, capturada na Arábia Saudita. Cinco de dez espécimes de víbora examinadas (50%) foram encontradas infectadas com Hepatozoon pyramidumi sp. n. com nível de parasitemia de 20% a 30%. A infecção foi restrita apenas aos eritrócitos. Foram observadas duas formas morfologicamente diferentes de estágios intra-eritrocíticos: gamontes de tamanho pequeno e madura. As formas menores de gamontes apresentaram média de 10,7 × 3,5 μm. Os gamontes maduros apresentaram forma de salsicha, com pequenos polos recurvados, medindo 16,3 × 4,2 μm, em média. Os eritrócitos infectados estavam aumentados de tamanho; seus núcleos encontravam-se deformados e, algumas vezes, deslocados de sua posição central, quando comparados às células normais não-infectadas. Foram observados estágios merogônicos em células endoteliais pulmonares e nas células do parênquima hepático. Os merontes maduros apresentavam 17,8 × 13,6 µm e continham merozoítos em forma de banana com tamanho médio de ~ 15 × 2 µm. A análise filogenética baseada nas sequências SSU rDNA agrupou Hepatozoon pyramidumi sp. n com Hepatozoon spp. detectados em répteis de várias regiões geográficas. Por meio de análises morfológicas e moleculares com espécies intimamente relacionadas, demonstrou-se a singularidade dessa nova espécie de Hepatozoon.(AU)
Assuntos
Animais , Elapidae/classificação , Elapidae/genética , Filogenia , ApicomplexaRESUMO
Abstract Hepatozoon pyramidumi sp. n. is described from the blood of the Egyptian saw-scaled viper, Echis pyramidum, captured from Saudi Arabia. Five out of ten viper specimens examined (50%) were found infected with Hepatozoon pyramidumi sp. n. with parasitaemia level ranged from 20-30%. The infection was restricted only to the erythrocytes. Two morphologically different forms of intraerythrocytic stages were observed; small and mature gamonts. The small ganomt with average size of 10.7 × 3.5 μm. Mature gamont was sausage-shaped with recurved poles measuring 16.3 × 4.2 μm in average size. Infected erythrocytes were hypertrophied; their nuclei were deformed and sometimes displaced from their central position in the normal uninfected cell. Merogonic stages were observed in the lung endothelial cell and the liver parenchyma cells. Mature meront was 17.8 × 13.6 µm and contained banana-shaped merozoites with average size of ~15 × 2 µm. Phylogenetic analysis based on the SSU rDNA sequence clustered Hepatozoon pyramidumi sp. n with previously sequenced Hepatozoon spp., most of them infected reptilian hosts without geographic consideration. The morphological and molecular comparison with closely related species proved the taxonomic uniqueness and novelty of the present form.
Resumo Hepatozoon pyramidumi sp. n. é descrito a partir do sangue da víbora em escamas e quilhas serrilhadas, Echis pyramidum, capturada na Arábia Saudita. Cinco de dez espécimes de víbora examinadas (50%) foram encontradas infectadas com Hepatozoon pyramidumi sp. n. com nível de parasitemia de 20% a 30%. A infecção foi restrita apenas aos eritrócitos. Foram observadas duas formas morfologicamente diferentes de estágios intra-eritrocíticos: gamontes de tamanho pequeno e madura. As formas menores de gamontes apresentaram média de 10,7 × 3,5 μm. Os gamontes maduros apresentaram forma de salsicha, com pequenos polos recurvados, medindo 16,3 × 4,2 μm, em média. Os eritrócitos infectados estavam aumentados de tamanho; seus núcleos encontravam-se deformados e, algumas vezes, deslocados de sua posição central, quando comparados às células normais não-infectadas. Foram observados estágios merogônicos em células endoteliais pulmonares e nas células do parênquima hepático. Os merontes maduros apresentavam 17,8 × 13,6 µm e continham merozoítos em forma de banana com tamanho médio de ~ 15 × 2 µm. A análise filogenética baseada nas sequências SSU rDNA agrupou Hepatozoon pyramidumi sp. n com Hepatozoon spp. detectados em répteis de várias regiões geográficas. Por meio de análises morfológicas e moleculares com espécies intimamente relacionadas, demonstrou-se a singularidade dessa nova espécie de Hepatozoon.
Assuntos
Animais , DNA de Protozoário/genética , Apicomplexa/fisiologia , Apicomplexa/genética , Viperidae/parasitologia , Filogenia , Arábia Saudita , DNA Ribossômico/genética , Apicomplexa/classificação , Análise de Sequência de DNA , Viperidae/sangue , Parasitemia/parasitologia , Parasitemia/veterinária , Eritrócitos , Eritrócitos/patologia , Fígado/parasitologia , Fígado/patologia , Pulmão/parasitologia , Pulmão/patologiaRESUMO
Based on morphology and morphometry of gametocytes in blood and molecular phylogenetic analysis, we described a new species of hemoparasite from the genus Haemogregarina isolated from Lepidosiren paradoxa in the eastern Amazon region. Haemogregarina daviesensis sp. nov. is characterized by monomorphic gametocytes of varying maturity stage and their dimensions were 16 ± 0.12 µm (range 13-18) in length and 6 ± 0.97 µm (range 5-8) in width. The morphological and morphometric data were not identical with other haemogregarine species from fish. All specimens of L. paradoxa analyzed were infected by H. daviesensis sp. nov. and the parasitemia level was moderate (1-28/2000 blood erythrocytes). Two sequences were obtained from L. paradoxa, and these constituted a monophyletic sister clade to the Haemogregarina species. In addition, H. daviesensis sp. nov. detected here grouped with Haemogregarina sp. sequences isolated from chelonian Macrochelys temminckii, with 99% bootstrap support. This study provides the first data on the molecular phylogeny of an intraerythrocytic haemogregarine of freshwater fish and highlights the importance of obtaining additional information on aspects of the general biology of these hemoparasites in fish populations, in order to achieve correct taxonomic classification.
Assuntos
Coccidiose/veterinária , Eucoccidiida/isolamento & purificação , Doenças dos Peixes/parasitologia , Animais , Coccidiose/parasitologia , Eucoccidiida/classificação , Eucoccidiida/genética , Eucoccidiida/fisiologia , Feminino , Peixes/parasitologia , Água Doce/parasitologia , Masculino , Parasitemia/parasitologia , Parasitemia/veterinária , FilogeniaRESUMO
Hepatozoon species are vector-borne pathogens that infect domestic and wild animals. Marsupials of the species Didelphis albiventris are adapted to urban and peri-urban areas and act as reservoir hosts for several parasites. The present study evaluated the occurrence of infection by Hepatozoon species in synantropic D. albiventris from Botucatu, São Paulo, Brazil. Blood samples and ectoparasites from 19 D. albiventris were collected from urban and peri-urban areas. Hepatozoon spp. detection was performed by microscopy and molecular analysis. One opossum was positive for Hepatozoon spp. in microscopy analysis and PCR, while another animal was positive only in PCR. The obtained sequences were 100% identical to Hepatozoon canis. Six species of ticks and two species of fleas were detected on D. albiventris. This is the first report of H. canis in synantropic D. albiventris. In Brazil, H. canis transmission among dog populations is not well established, which highlights the importance of investigating the role that opossums might play in the epidemiology of this protozoan.
Assuntos
Coccidiose/veterinária , Didelphis , Eucoccidiida/isolamento & purificação , Infestações por Pulgas/veterinária , Infestações por Carrapato/veterinária , Animais , Brasil/epidemiologia , Coccidiose/epidemiologia , Coccidiose/parasitologia , Ctenocephalides/classificação , Eucoccidiida/classificação , Eucoccidiida/genética , Infestações por Pulgas/epidemiologia , Infestações por Pulgas/parasitologia , Ixodidae/classificação , Ixodidae/crescimento & desenvolvimento , Larva/classificação , Larva/crescimento & desenvolvimento , Ninfa/classificação , Ninfa/crescimento & desenvolvimento , Ornithodoros/classificação , Ornithodoros/crescimento & desenvolvimento , Filogenia , Análise de Sequência de DNA , Sifonápteros/classificação , Infestações por Carrapato/epidemiologia , Infestações por Carrapato/parasitologiaRESUMO
To date, only two haemogregarine parasite species have been described from South African anurans: Hepatozoon ixoxo, infecting toads of the genus Sclerophrys (syn. Amietophrynus); and Hepatozoon theileri, parasitising the common river frog, Amietia quecketti. Both species have been characterised using limited morphology, and molecular data from PCR amplified fragments of the 18S rRNA gene. However, no ultrastructural work has been performed thus far. The aim of this study was to add descriptive information on the two species by studying their ultrastructural morphology. Mature gamont stages, common in the peripheral blood of infected frogs, were examined by transmission electron microscopy. Results indicate that H. ixoxo and H. theileri share typical apicomplexan characteristics, but differ markedly in their external cellular structure. Hepatozoon ixoxo is an encapsulated parasite presenting a prominent cap at the truncate pole, and shows no visible modifications to the host cell membrane. In comparison, H. theileri does not present a capsule or cap, and produces marked morphological changes to its host cell. Scanning electron microscopy was performed to further examine the cytopathological effects of H. theileri, and results revealed small, knob-like protrusions on the erythrocyte surface, as well as notable distortion of the overall shape of the host cell.
Assuntos
Anuros/parasitologia , Coccidiose/parasitologia , Coccidiose/veterinária , Eucoccidiida/ultraestrutura , Doenças dos Animais/parasitologia , Animais , Coccidiose/sangue , DNA de Protozoário/genética , Eritrócitos/parasitologia , Eucoccidiida/classificação , Eucoccidiida/genética , Microscopia Eletrônica de Varredura , Microscopia Eletrônica de Transmissão , Filogenia , RNA Ribossômico 18S/genética , Rios , América do Sul , Especificidade da EspécieRESUMO
A haemogregarine is described in 12 cururu stingray (Potamotrygon cf. histrix), from Mariuá Archipelago, Negro River, in the Brazilian Amazon Basin. All animals, both male and female, were parasitized by the haemogregarine and parasitaemia varied between 0.8% and 10% of erythrocytes. The stages observed included trophozoites or merozoites, suspected meronts, and gamonts presumed to be of two types, macrogamonts and microgamonts. Most stages were observed inside mature erythrocytes, while others were extracellular. The stages observed were most similar to those characteristics of the genus Cyrilia, than to any other fish haemogregarine and may represent a new Cyrilia species.
Assuntos
Coccidiose/veterinária , Eucoccidiida/classificação , Eucoccidiida/fisiologia , Doenças dos Peixes/parasitologia , Parasitemia/veterinária , Rajidae , Animais , Brasil/epidemiologia , Coccidiose/epidemiologia , Coccidiose/parasitologia , Eritrócitos/parasitologia , Eucoccidiida/citologia , Feminino , Doenças dos Peixes/epidemiologia , Masculino , Parasitemia/epidemiologia , Parasitemia/parasitologia , Prevalência , Rios , Especificidade da EspécieRESUMO
Twenty-five black river turtles (Rhinoclemmys funerea) and eight white-lipped mud turtles (Kinosternon leucostomum) from Selva Verde, Costa Rica were examined for haemoparasites. Leeches identified as Placobdella multilineata were detected on individuals from both species. All turtles sampled were positive for intraerythrocytic haemogregarines (Apicomplexa:Adeleorina) and the average parasitemia of black river turtles (0.34% ± 0.07) was significantly higher compared to white-lipped mud turtles (0.05% ± 0.006). No correlation was found between parasitemia and relative body mass of either species or between black river turtles from the two habitats. In addition, one scorpion mud turtle (Kinosternon scorpioides) examined from La Pacifica, Costa Rica, was positive for haemogregarines (0.01% parasitemia). Interestingly, parasites of the scorpion mud turtle were significantly smaller than those from the other two species and did not displace the erythrocyte nucleus, whereas parasites from the other two species consistently displaced host cell nuclei and often distorted size and shape of erythrocytes. This is the first report of haemogregarines in turtles from Central America and of haemogregarines in K. leucostomum, K. scorpioides, and any Rhinoclemmys species. Additional studies are needed to better characterise and understand the ecology of these parasites.