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1.
Parasitol Res ; 119(9): 3023-3031, 2020 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-32725320

RESUMO

Hepatozoon canis is a hemoprotozoan organism that infects domestic and wild carnivores throughout much of Europe. The parasite is mainly transmitted through the ingestion of infected ticks containing mature oocysts. The aims of the present survey were to determine the prevalence of H. canis in hunting dogs living in Southern Italy and to assess potential infection risk factors. DNA extracted from whole blood samples, collected from 1433 apparently healthy dogs living in the Napoli, Avellino, and Salerno provinces of Campania region (Southern Italy), was tested by a quantitative real-time polymerase chain reaction (qPCR) assay to amplify H. canis. Furthermore, the investigated dog population was also screened by qPCR for the presence of Ehrlichia canis, a major tick-borne pathogen in Southern Italy, in order to assess possible co-infections. Two hundred dogs were H. canis PCR-positive, resulting in an overall prevalence of 14.0% (CI 12.2-15.9). Breed category (P < 0.0001), hair coat length (P = 0.015), and province of residence (P < 0.0001) represented significant risk factors for H. canis infection. The presence of H. canis DNA was also significantly associated with E. canis PCR positivity (P < 0.0001). Hunting dogs in Campania region (Southern Italy) are frequently exposed to H. canis, and the infection is potentially associated with close contact with wildlife. Further studies are needed to assess the pathogenic potential of H. canis, as well as the epidemiological relationships between hunting dogs and wild animal populations sharing the same habitats in Southern Italy.


Assuntos
Coccidiose/veterinária , Doenças do Cão/parasitologia , Eucoccidiida/isolamento & purificação , Animais , Coccidiose/parasitologia , Coccidiose/transmissão , Doenças do Cão/sangue , Doenças do Cão/epidemiologia , Doenças do Cão/transmissão , Cães , Eucoccidiida/genética , Eucoccidiida/fisiologia , Feminino , Itália/epidemiologia , Masculino , Reação em Cadeia da Polimerase , Fatores de Risco , Carrapatos/parasitologia , Carrapatos/fisiologia
2.
Parasitol Res ; 119(10): 3315-3326, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32699938

RESUMO

Haemogregarina species are apicomplexan blood parasites infecting vertebrates such as fish, lizards, and turtles. Due to the high morphological similarity of the erythrocytic stages infecting host species, it has always been a challenge to identify the true diversity of these parasites. Therefore, taxonomic studies are presently based on the combination of morphological and molecular data. In Tunisia, two species of Haemogregarina have been reported within the freshwater turtle Mauremys leprosa (Geoemydidae) for more than 40 years. Since M. leprosa occurs in the same aquatic environments as Emys orbicularis (Emydidae) in Tunisia, our objectives were to assess parasite diversity and specificity on the basis of both morphological and molecular approaches. The turtles were surveyed and sampled across six aquatic areas of Tunisia. Among the 39 specimens of M. leprosa and seven of E. orbicularis that were trapped and investigated, the presence of haemogregarines was detected in the blood of turtles only at sites where leeches were observed. Three 18S variants were identified, which corresponded to three distinct Haemogregarina species, among which one was identified as Haemogregarina stepanowi. The two other species that were detected are likely new to science. Because we show the occurrence of more than one blood parasite species within a single host specimen, our study provides the first report of coinfection with molecularly distinct Haemogregarina spp.


Assuntos
Coccidiose/veterinária , Coinfecção/veterinária , Eucoccidiida/genética , Água Doce/parasitologia , Tartarugas/parasitologia , Animais , Coccidiose/parasitologia , Coinfecção/parasitologia , Eucoccidiida/classificação , Eucoccidiida/isolamento & purificação , Variação Genética , Sanguessugas/parasitologia , RNA Ribossômico 18S/genética , Tunísia/epidemiologia , Tartarugas/classificação
3.
Parasitol Res ; 119(6): 1891-1901, 2020 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-32382990

RESUMO

Amphibian populations are declining around the world, and the main reasons are the environmental changes and pathogens. However, there are few studies addressing the interaction and impact of the different pathogens that affect amphibians, such as hemoparasites. These parasites had been described as common in some amphibian species, but unfortunately, their description and characterization are unclear and scarcely spread. The objective of the present study was to evaluate the morphological and molecular characterization of hemoparasites present in vaillant's frogs. Seven frogs of Lithobates vaillanti were captured at the biological station La Florida in Tabasco, Mexico. Blood smears were performed, and results show that 100% of the animals have hemoparasites. Three types of hemoparasites were found. Eighty-five percent of the frogs were positive to Hepatozoon sp., 57% to Lankesterella sp., and 28% to Trypanosoma sp. According to the molecular analysis of the obtained sequences of Trypanosoma sp. and Hepatozoon sp., both protozoans were positioned in between the clusters of parasites of different geographical regions. Nevertheless, no species names were assigned to any of these parasites because more sequences and analysis are needed.


Assuntos
Ranidae/parasitologia , Animais , Eucoccidiida/classificação , Eucoccidiida/isolamento & purificação , Florida , México , Parasitos/classificação , Parasitos/isolamento & purificação , Trypanosoma/classificação , Trypanosoma/isolamento & purificação
4.
Rev Bras Parasitol Vet ; 29(1): e018019, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32049147

RESUMO

The aim of the present study was to detect Cercopithifilaria bainae and other tick-borne pathogens and to perform molecular characterization of the tick Rhipicephalus sanguineus s.l. collected from dogs. Ticks (n = 432, including 8 larvae, 59 nymphs, and 365 adults) were sampled from domiciled dogs (n = 73) living in Campo Grande, Mato Grosso do Sul (Midwest Brazil). All ticks were morphologically identified as R. sanguineus. Genomic DNA was extracted in pools (three to five ticks per animal) and was used for definition of R. sanguineus haplotypes (based on 16S rRNA analysis) and pathogen identification (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli and Rickettsia spp.). Rhipicephal us sanguineus specimens were identified as haplotypes A and B. DNA of Cercopithifilaria bainae (43.83%; 32/73), Ehrlichia canis (24.65%; 18/73), Anaplasma platys (19.17%; 14/73), and Hepatozoon canis (5.47%; 4/73) was detected. The identity of pathogens was confirmed by DNA sequence analysis. The present study confirms the presence of haplotypes A and B of R. sanguineus in the state of Mato Grosso do Sul and its importance as a vector of several pathogens of veterinary concern. Finally, this is the first report to identify C. bainae in ticks in the Midwestern region of Brazil.


Assuntos
Vetores Aracnídeos/parasitologia , Cães/parasitologia , Rhipicephalus sanguineus/parasitologia , Anaplasma/genética , Anaplasma/isolamento & purificação , Animais , Babesia/genética , Babesia/isolamento & purificação , Brasil , Ehrlichia canis/genética , Ehrlichia canis/isolamento & purificação , Eucoccidiida/genética , Eucoccidiida/isolamento & purificação , RNA Ribossômico 16S/genética , Rickettsia/genética , Rickettsia/isolamento & purificação
5.
Parasitol Res ; 119(2): 713-724, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31912276

RESUMO

Intra-leukocytic gamonts consistent with the description of Hepatozoon griseisciuri Clark, 1958 are reported for the first time in Canadian eastern gray squirrels (Sciurus carolinensis Gmelin, 1788). Polymerase chain reaction (PCR) amplification and direct Sanger sequencing identified a pair of distinct genotypes at both a nuclear and mitochondrial locus; two 18S ribosomal RNA gene sequences (rDNA; genotype A and genotype B: 1816 base pairs (bp); 98.8% pairwise identity) and 2 distinct complete mitochondrial genome sequences (genotype A: 6311 bp; genotype B: 6114 bp; 89.1% pairwise identity) were obtained from 3 H. griseisciuri-infected squirrels sampled in Guelph, Ontario. The genetic content of both circular-mapping mitochondrial genomes was conventional for apicomplexan protists; each encoded for 3 protein-coding genes (cytochrome c oxidase subunit I (COI); cytochrome c oxidase subunit III (COIII); and cytochrome B (CytB)), 14 fragmented large subunit rDNA, 10 fragmented small subunit rDNA, and 8 unassigned rDNA. These genotypes, based on sequences obtained from a pair of loci from two parasite genomes, confirm the presence of at least two Hepatozoon species infecting Ontario eastern gray squirrels, one of which is likely to be conspecific with H. griseisciuri.


Assuntos
Eucoccidiida/classificação , Eucoccidiida/genética , Sciuridae/parasitologia , Animais , Citocromos b/genética , DNA Ribossômico/genética , Complexo IV da Cadeia de Transporte de Elétrons/genética , Eucoccidiida/isolamento & purificação , Genoma Mitocondrial/genética , Genótipo , Tipagem de Sequências Multilocus , Ontário , Filogenia , RNA Ribossômico 18S/genética
6.
Parasitol Res ; 119(2): 687-694, 2020 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-31897793

RESUMO

Wild rodents, as natural reservoir hosts carrying various species of pathogens, play an important role in the evolution and emergence of zoonotic diseases. In this study, protist parasites, namely Babesia sp., Trypanosoma sp. and Hepatozoon sp. were studied in rodent populations in Lithuania. Two hundred forty rodent specimens of seven species were analysed by a combined approach using polymerase chain reaction (PCR)-based techniques and traditional microscopic examination. The total prevalence of blood parasites reached 35% in rodent communities. The prevalence of Hepatozoon sp. reached the highest value (32%), followed by Trypanosoma sp. (5%) and Babesia sp. (3%). Myodes glareolus and Microtus agrestis were the most heavily infected rodent species. Comparison of microscopy and PCR-based methods showed that the two approaches might give different results and thus can lead to an underestimation of the actual prevalence and abundance of parasites. In our study, PCR-based assays were more sensitive and robust than traditional microscopy. However, precise molecular results for the estimation of the prevalence of Babesia sp. and Hepatozoon sp. were achieved only by using several sets of primers. To avoid inaccurate results, the improvement and detailed description of molecular and microscopy protocols are required.


Assuntos
Arvicolinae/parasitologia , Babesia/isolamento & purificação , Eucoccidiida/isolamento & purificação , Trypanosoma/isolamento & purificação , Animais , Lituânia , Microscopia , Reação em Cadeia da Polimerase
7.
Parasitol Res ; 119(3): 1093-1100, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31938888

RESUMO

The purpose of this study was to perform a phylogenetic analysis of Hepatozoon spp. infecting Philodryas patagoniensis in Uruguay. Twenty-five road-killed specimens of P. patagoniensis from ten departments were obtained. Samples of blood and/or heart tissue were taken. Polymerase chain reaction (PCR) assay was carried out amplifying a specific target region of the 18S rRNA gene of Hepatozoon spp. Eighteen out of twenty-five samples were positive to Hepatozoon spp., which gave an overall prevalence of 72%. Phylogenetic analyses with the obtained sequences were carried out to determine the relationship with closely related species found in the region. The results revealed that samples were split into two clades with a high bootstrap support. Clade I was formed with Hepatozoon spp. sequences obtained in this study from P. patagoniensis, Hepatozoon cuestensis from Crotalus durissus terrificus and Hepatozoon musa from Philodryas nattereri, and Hepatozoon spp. retrieved from Cerdocyon thous, Hemidactylus mabouia, and Phyllopezus pollicaris from Brazil, respectively. Clade II was grouped with Hepatozoon cevapii and Hepatozoon massardii, both species described for C. d. terrificus from Brazil. This is the first report of Hepatozoon spp. in snakes from Uruguay.


Assuntos
Coccidiose/veterinária , Colubridae/parasitologia , Eucoccidiida/classificação , Eucoccidiida/isolamento & purificação , Filogenia , Animais , Coccidiose/epidemiologia , Coccidiose/parasitologia , DNA de Protozoário/genética , Eucoccidiida/genética , Prevalência , RNA Ribossômico 18S/genética , Uruguai/epidemiologia
8.
Acta Parasitol ; 65(1): 151-155, 2020 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-31729613

RESUMO

BACKGROUND: Laboratory diagnosis of Hepatozoon canis infection is tedious, especially in chronic and/or latent infections. PURPOSE: The study was planned to develop a simple read out loop mediated isothermal amplification (LAMP) assay targeting a partial 18S rRNA gene of H. canis with naked eye visualisation of LAMP products. METHODS: A LAMP assay was employed to assess the DNA amplification by adding SYBR Green I dye for naked eye inspection of DNA accumulating in reaction tubes. Positive amplification was read through observation of change in colour of reaction mixture following addition of dye. The visual results were further verified with those of agarose gel electrophoresis. Genomic DNA of other haemoparasites of dog viz. Babesia vogeli, B. gibsoni, Ehrlichia canis and Trypanosoma evansi along with no-template control were used to determine the specificity of assay. RESULTS: Among the 109 blood samples presented at Small Animal Clinics, Teaching Veterinary Clinical Complex, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab (India) tested, 39 revealed colour change from orange to green indicating positive reaction while 70 were negative as revealed by no colour change. The results of visual inspection were comparable to those obtained by agarose gel electrophoresis. The LAMP primers specifically amplified H. canis DNA, whereas no amplification was detected in DNA samples of other haemoparasites and no-template control revealing specificity of the assay. The diagnostic sensitivity and specificity (95% CI) of visual LAMP assay with respect to microscopy in detection of H. canis varied from 100% (15.81-100.00%) and 65.42% (55.61-74.35%), respectively. CONCLUSION: The present investigation has developed a specific and rapid LAMP assay for the detection of H. canis, using SYBR Green I dye, which has practical applications for the screening of field samples.


Assuntos
Coccidiose/veterinária , Doenças do Cão/diagnóstico , Eucoccidiida/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/veterinária , Temperatura , Animais , Coccidiose/diagnóstico , Primers do DNA/genética , Doenças do Cão/parasitologia , Cães , Índia , Compostos Orgânicos , RNA Ribossômico 18S/genética , Sensibilidade e Especificidade
9.
Rev. bras. parasitol. vet ; 29(1): e018019, 2020. tab, graf
Artigo em Inglês | LILACS | ID: biblio-1058020

RESUMO

Abstract The aim of the present study was to detect Cercopithifilaria bainae and other tick-borne pathogens and to perform molecular characterization of the tick Rhipicephalus sanguineus s.l. collected from dogs. Ticks (n = 432, including 8 larvae, 59 nymphs, and 365 adults) were sampled from domiciled dogs (n = 73) living in Campo Grande, Mato Grosso do Sul (Midwest Brazil). All ticks were morphologically identified as R. sanguineus. Genomic DNA was extracted in pools (three to five ticks per animal) and was used for definition of R. sanguineus haplotypes (based on 16S rRNA analysis) and pathogen identification (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli and Rickettsia spp.). Rhipicephal us sanguineus specimens were identified as haplotypes A and B. DNA of Cercopithifilaria bainae (43.83%; 32/73), Ehrlichia canis (24.65%; 18/73), Anaplasma platys (19.17%; 14/73), and Hepatozoon canis (5.47%; 4/73) was detected. The identity of pathogens was confirmed by DNA sequence analysis. The present study confirms the presence of haplotypes A and B of R. sanguineus in the state of Mato Grosso do Sul and its importance as a vector of several pathogens of veterinary concern. Finally, this is the first report to identify C. bainae in ticks in the Midwestern region of Brazil.


Resumo O objetivo do presente estudo foi detectar Cercopithifilaria bainae e outros patógenos transmitidos por carrapatos e realizar a caracterização molecular do carrapato Rhipicephalus sanguineus s.l. coletado em cães. Carrapatos (n = 432, incluindo 8 larvas, 59 ninfas e 365 adultos) foram amostrados de cães domiciliados (n = 73) residentes no município de Campo Grande, Mato Grosso do Sul (centro-oeste do Brasil). Todos os carrapatos foram identificados morfologicamente como R. sanguineus. O DNA genômico foi extraído em pools (três a cinco carrapatos por animal), seguido pela definição de haplótipos (com base no gene 16S rRNA) e pela investigação de patógenos (Cercopithifilaria sp., Ehrlichia canis, Anaplasma platys, Hepatozoon canis, Babesia vogeli e Rickettsia spp.). Os espécimes coletados foram identificados como haplótipos A e B de R. sanguineus. Foram detectados DNA de Cercopithifilaria bainae (43,83%; 32/73), Ehrlichia canis (24,65%; 18/73), Anaplasma platys (19,17%; 14/73) e Hepatozoon canis (5,47%; 4/73). A identidade dos patógenos foi confirmada por análise de sequência de DNA. O presente estudo confirma a circulação dos haplótipos A e B de R. sanguineus no estado de Mato Grosso do Sul e sua importância como vetor de vários patógenos de interesse veterinário. Finalmente, este é o primeiro relato de C. bainae em carrapatos na região centro-oeste do Brasil.


Assuntos
Animais , Vetores Aracnídeos/parasitologia , Rhipicephalus sanguineus/parasitologia , Cães/parasitologia , Rickettsia/isolamento & purificação , Rickettsia/genética , Babesia/isolamento & purificação , Babesia/genética , Brasil , RNA Ribossômico 16S/genética , Eucoccidiida/isolamento & purificação , Eucoccidiida/genética , Ehrlichia canis/isolamento & purificação , Ehrlichia canis/genética , Anaplasma/isolamento & purificação , Anaplasma/genética
10.
Exp Appl Acarol ; 79(1): 137-155, 2019 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-31489558

RESUMO

The objective of the present study was to detect the chosen nucleotide DNA or RNA sequences of the pathogens in ticks of domestic and wild animals of Kerala, South India based on molecular techniques. Among 602 ticks collected, 413 were from bovines (cattle and buffalo), 26 from goats, 101 from dogs and 62 from wild animals. Amblyomma integrum, Am. gervaisi, Dermacentor auratus, Haemaphysalis bispinosa, Ha. intermedia, Ha. shimoga, Ha. spinigera, Rhipicephalus annulatus, Rh. microplus, Rh. haemaphysaloides and Rh. sanguineus s.l. were identified from various domestic and wild animals of Kerala. The cDNA synthesized from the RNA isolated from fully or partially engorged adult female/nymphal ticks was used as template for the specific polymerase chain reactions (PCR). Out of 602 ticks examined, nucleotide sequences of pathogens were detected in 28 ticks (4.65%). The nucleotide sequences of tick-borne pathogens like Theileria orientalis, Babesia vogeli, Hepatozoon canis, Anaplasma marginale, An. bovis, Rickettsia sp. closely related to Ri. raoultii, Ri. massiliae, Ri. africae and Ri. slovaca were detected. The identification of the previously unreported nucleotide sequences of rickettsial pathogens from India is of particular interest due to their zoonotic significance. The phylogenetic analysis of the major piroplasm surface protein (MPSP) gene of T. orientalis amplified from Rh. annulatus ticks revealed that they were genetically close to type 7, which belong to the highly pathogenic Ikeda group.


Assuntos
Animais Selvagens , Eucoccidiida/isolamento & purificação , Interações Hospedeiro-Parasita , Ixodidae , Piroplasmida/isolamento & purificação , Rickettsiales/isolamento & purificação , Infestações por Carrapato/veterinária , Animais , Índia , Ixodidae/microbiologia , Ixodidae/parasitologia , Ixodidae/fisiologia , Filogenia , Infestações por Carrapato/parasitologia
11.
Parasitol Res ; 118(10): 2773-2779, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31456045

RESUMO

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 , Filogenia
12.
Parasitol Int ; 73: 101975, 2019 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-31421267

RESUMO

This work reports the occurrence of coccidia of the genus Calyptospora in fishes from the eastern Amazon. Fish were collected on flood plains in the municipality of Macapá, State of Amapá, Brazil. Fresh squash preparations of liver, heart and gallbladder were examined under light microscope. Positive samples of Geophagus proximus and Hoplias malabaricus were used to detect parasites by PCR with Calyptospora-specific primers mRF and mrR, which amplify a region of the 18S rRNA gene. Oocysts were observed in 55% of 130 fishes examined. Parasite prevalence varied according to feeding habits, and was 100% in carnivores, 74% in omnivores (invertivores and detritivores) and 0% in herbivores. Variation in the frequency of parasitized organs showed 100% in the liver, 30% in the gallbladder, and 9% in the heart. The sequences obtained from G. proximus and H. malabaricus were identical and showed 99% similarity to Calyptospora serrasalmi. To our knowledge, this is the first report of the occurrence of Calyptospora in 10 new species of fish from the region of the eastern Brazilian Amazon. The results demonstrate the occurrence of C. serrasalmi in the region and the research provides new primers for the diagnosis of Calyptospora spp.


Assuntos
Coccidiose/veterinária , Eucoccidiida/isolamento & purificação , Doenças dos Peixes/epidemiologia , Peixes , Animais , Brasil , Caraciformes , Ciclídeos , Coccidiose/epidemiologia , Coccidiose/parasitologia , Dieta , Doenças dos Peixes/parasitologia , Reação em Cadeia da Polimerase/veterinária , Prevalência , RNA de Protozoário/análise , RNA Ribossômico 18S/análise
13.
Parasit Vectors ; 12(1): 294, 2019 Jun 11.
Artigo em Inglês | MEDLINE | ID: mdl-31186033

RESUMO

BACKGROUND: Vector-borne diseases are emerging worldwide and have an important zoonotic relevance. In the last few years, the interest in vector-borne pathogens in cats has increased. However, studies on feline vector-borne pathogens on tropical islands are lacking. Islands differ from continental countries because they have an enclosed population of animals, with all year presence of the vectors and, most often, without vector control measures. This study focused on the molecular identification and phylogenetic analysis of vector-borne pathogens in autochthonous cats with a mixed indoor-outdoor lifestyle from Maio Island, Cape Verde archipelago. METHODS: Blood samples were collected from 80 asymptomatic cats, representing almost a quarter of the total cat population of the island. The presence of DNA of protozoa of the genus Hepatozoon and bacteria belonging to family Anaplasmataceae and to genus Bartonella was assessed by PCR and phylogenetic analysis was conducted. Statistical analysis was performed to identify risk factors associated with infection. For feline hepatozoonosis, a worldwide dataset of Hepatozoon felis sequences retrieved from mammal species and vectors along with Hepatozoon spp. sequences retrieved from felids was generated, phylogenetically analyzed and the geographical and host distribution was assessed. RESULTS: Hepatozoon felis genotype I was identified in 12 (15%) cats from Maio Island whereas none of the cats were PCR positive for the other pathogens tested. No significant association of H. felis infection with age, sex, location or presence of vectors was observed by statistical analysis in Cape Verde's cats. Phylogenetic analysis on the worldwide dataset of feline Hepatozoon sequences showed two significant distinct clades for H. felis genotype I and II. Different geographical distributions were assessed: H. felis genotype I was the only genotype found in Africa and has been reported worldwide, with the exception of Japan and Brazil where only H. felis genotype II has been reported. CONCLUSIONS: The identification of H. felis genotype I in cats in Maio Island highlights the need to further investigate the significance of H. felis genotypes and to clarify the epidemiological aspects of this infection.


Assuntos
Doenças do Gato/epidemiologia , Coccidiose/veterinária , Eucoccidiida/genética , Genótipo , Anaplasma/genética , Animais , Infecções Assintomáticas , Bartonella/genética , Cabo Verde/epidemiologia , Doenças do Gato/parasitologia , Gatos/parasitologia , Coccidiose/epidemiologia , DNA de Protozoário/genética , Vetores de Doenças , Eucoccidiida/isolamento & purificação , Eucoccidiida/patogenicidade , Ilhas , Filogenia , Fatores de Risco , Carrapatos/parasitologia , Clima Tropical
14.
Parasitol Int ; 72: 101945, 2019 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-31228586

RESUMO

Three different Hepatozoon (Apicomplexa, Hepatozoidae) species have been described infecting domestic cats in Europe (i.e. H. felis, H. canis and H. silvestris), however, reports on clinical hepatozoonosis are uncommon and treatment protocols are not clearly defined. A six-year-old male European short-hair cat from Austria presented poor general condition, lethargy, anorexia, icterus, a painful abdomen, fever, ruffled hair and a tick infestation, and it had never left Austria. Laboratory tests revealed leukopenia, thrombocytopenia and increased serum levels of symmetric dimethylarginine (SDMA) and bilirubin. In May Grünwald-Giemsa-stained blood smears, structures resembling Hepatozoon gamonts were observed inside neutrophil granulocytes. A PCR targeting a fragment of the 18S rRNA gene of Hepatozoon spp. and DNA sequencing allowed the diagnosis of H. felis-DNA in blood samples. The cat was treated with imidocarb dipropionate (6 mg/kg body weight, repeated after 14 days) and doxycycline monohydrate (5 mg/kg body weight twice a day, p.o., for four weeks) and recovered completely. A broad haematological and biochemical laboratory control after six months showed all evaluated parameters under normal ranges. Coinfection with other feline pathogens (i.e. feline leukaemia virus, feline immunodeficiency virus, feline Coronavirus, Leishmania and Dirofilaria immitis) could not be detected. This study reveals the presence of H. felis in Austria and provides more evidence on the geographical distribution and pathogenicity of this parasite for domestic cats. To the authors' knowledge, this is the first autochthonous case of feline hepatozoonosis in Central Europe.


Assuntos
Doenças do Gato/diagnóstico , Doenças do Gato/parasitologia , Gatos/parasitologia , Coccidiose/veterinária , Eucoccidiida/isolamento & purificação , Animais , Antiprotozoários/uso terapêutico , Áustria , Doenças do Gato/tratamento farmacológico , Coccidiose/diagnóstico , Eucoccidiida/genética , Imidocarbo/uso terapêutico , Masculino , Reação em Cadeia da Polimerase , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Resultado do Tratamento
15.
Ticks Tick Borne Dis ; 10(4): 929-934, 2019 06.
Artigo em Inglês | MEDLINE | ID: mdl-31078466

RESUMO

We determined the prevalence of infection and genetic identity of Hepatozoon spp. harbored by Rhipicephalus sanguineus sensu lato ticks in Taiwan. A total of 1082 ticks were collected from dogs and DNA extraction was performed from individual tick specimens. Hepatozoon infection was detected by performing a nested-PCR assay based on the 18S small subunit ribosomal RNA (ssrRNA) gene. The genetic identity of detected Hepatozoon was identified by gene sequencing and phylogenetic analysis. Hepatozoon infection was detected in nymphs, males and females of R. sanguineus s. l. ticks with an infection rate of 20.8%, 22.8% and 29.4%, respectively. Sequence and phylogenetic analysis revealed that these Hepatozoon spp. of Taiwan were genetically affiliated to the same clade within the genospecies of H. canis and can be discriminated from other genospecies of H. americanum and H. felis. Intraspecies analysis based on the genetic distance (GD) values indicates a lower level (GD < 0.005) genetic divergence within the same genospecies of H. canis detected in Taiwan, Brazil and Spain. Interspecies analysis also reveals a higher heterogeneity of Taiwan strains distinguished from other genospecies of H. felis (GD > 0.040) and H. americanum (GD > 0.056). This study provides the first molecular evidence of H. canis detected and identified in various stages of R. sanguineus s. l. ticks in Taiwan. Detection of H. canis in unfed male ticks may imply the possible mechanism of transstadial survival in R. sanguineus s. l. ticks. Further investigations on Hepatozoon spp. harbored by various vector ticks in Taiwan may illustrate the epidemiological significance of this parasite.


Assuntos
Coccidiose/veterinária , Doenças do Cão/epidemiologia , Doenças do Cão/parasitologia , Cães/parasitologia , Eucoccidiida/genética , Rhipicephalus sanguineus/parasitologia , Animais , Vetores Aracnídeos/parasitologia , Brasil/epidemiologia , Coccidiose/epidemiologia , DNA de Protozoário/genética , Eucoccidiida/isolamento & purificação , Feminino , Masculino , Ninfa/parasitologia , Filogenia , Reação em Cadeia da Polimerase , Prevalência , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Espanha/epidemiologia , Taiwan/epidemiologia
16.
Acta Trop ; 193: 78-83, 2019 May.
Artigo em Inglês | MEDLINE | ID: mdl-30831114

RESUMO

Canine tick-borne diseases have been considered emerging and re-emerging threats, given their increasing global prevalence. In this molecular survey, we aimed to detect and identify common tick-borne pathogens in dogs from Riyadh city in Saudi Arabia. Initially, the study included 36 dogs visiting private veterinary clinics. PCRs targeting the 18S ribosomal RNA gene (rDNA) of haemoparasites (Babesia, Theileria and Hepatozoon) and the 16S rDNA of Anaplasmataceae were performed. The results showed that 26 (72.2%) dogs were infected by some of the haemoparasites under investigation. The sequencing analysis of the amplicons confirmed the infections due to two parasite species Theileria equi and Theileria velifera. Further examination of guard dogs kept in the horse stables of the Riyadh Municipality revealed that the majority of the tested dogs (65.2%: 30 out of 46) were infected with either of the parasites. In addition, the genotypes of all the parasites in these dogs were identical to those of the parasites in the dogs from the veterinary clinics. Thus, it can be concluded that dogs are infected with these haemoparasites and serve as a reservoir for both T. equi and T. velifera in the study area; however, the clinical implication of this finding is to be studied.


Assuntos
Doenças do Cão/parasitologia , Piroplasmida/isolamento & purificação , Infecções por Protozoários/parasitologia , Espécies Sentinelas , Doenças Transmitidas por Carrapatos/parasitologia , Anaplasmataceae/isolamento & purificação , Animais , Babesia/isolamento & purificação , Babesiose/epidemiologia , Bovinos , Doenças dos Bovinos/epidemiologia , Coccidiose/epidemiologia , Reservatórios de Doenças , Doenças do Cão/epidemiologia , Cães , Eucoccidiida/isolamento & purificação , Feminino , Doenças dos Cavalos/epidemiologia , Cavalos , Masculino , Infecções por Protozoários/epidemiologia , Arábia Saudita/epidemiologia , Theileria/isolamento & purificação , Theileriose/epidemiologia , Doenças Transmitidas por Carrapatos/epidemiologia
17.
Parasitology ; 146(6): 728-739, 2019 05.
Artigo em Inglês | MEDLINE | ID: mdl-30871644

RESUMO

Oceanic islands are hotspots of biodiversity due to their high levels of endemism, with the Canary Islands being a notable example. A previous molecular study on the biogeography and host associations of haemogregarines (Apicomplexa: Adeleorina) infecting lizards from this archipelago detected seven parasite haplogroups. These haplogroups exhibited high host-specificity and geographical structure, suggesting that they might correspond to distinct biological identities. In this study, along with sequencing a longer fragment of the 18S rRNA, we further explore the distinctiveness of these parasites by analysing their morphology, effects on host erythrocytes and parasitaemia levels. These lines of evidence together with their genetics, host associations, frequency of occurrence and geographical distribution support them as different biological entities. As such, we describe seven new species: Karyolysus canariensis sp. nov., Karyolysus galloti sp. nov., Karyolysus stehlini sp. nov., Karyolysus gomerensis sp. nov., Karyolysus atlanticus sp. nov., Karyolysus tinerfensis sp. nov. and Karyolysus makariogeckonis sp. nov. These new taxa are further examples of endemic diversity in the Canarian archipelago. They also contribute to clarify the taxonomy within the Apicomplexa, a phylum estimated to have one of the lowest percentages of described species.


Assuntos
Coccidiose/veterinária , Eucoccidiida/classificação , Eucoccidiida/isolamento & purificação , Lagartos/parasitologia , Animais , Análise por Conglomerados , Coccidiose/parasitologia , DNA de Protozoário/química , DNA de Protozoário/genética , DNA Ribossômico/química , DNA Ribossômico/genética , Eucoccidiida/genética , Haplótipos , Especificidade de Hospedeiro , Filogenia , RNA Ribossômico 18S/genética , Análise de Sequência de DNA , Espanha
18.
Ticks Tick Borne Dis ; 10(2): 371-376, 2019 02.
Artigo em Inglês | MEDLINE | ID: mdl-30503892

RESUMO

The laboratory diagnosis of canine hepatozoonosis, caused by Hepatozoon canis is tedious, especially in chronic and latent infections. In the present investigation, a loop mediated isothermal amplification (LAMP) assay was developed and standardized targeting the partial 18S rRNA gene (GenBank accession no. KU096058). The LAMP primers specifically amplified H. canis DNA, whereas no amplification was detected in DNA samples from dogs infected with Babesia vogeli, B. gibsoni, Ehrlichia canis and Trypanosoma evansi, and no amplification was observed in DNA samples from H. canis-free dogs. The threshold sensitivity level of the assay was determined to be 15 fg of genomic DNA of H. canis. Furthermore, evaluation of blood samples collected from 250 dogs presented at Small Animal Clinics, Teaching Veterinary Clinical Complex, Guru Angad Dev Veterinary and Animal Sciences University, Ludhiana, Punjab (India) was carried out for the presence of H. canis by microscopy, 18S PCR assay and LAMP assay. Of the total samples subjected to these tests, LAMP detected H. canis in 75 samples, while 18S PCR and microscopy detected H. canis in 28 and 9 samples, respectively. The present investigation has developed, for the first time, a highly sensitive, specific and rapid LAMP assay for the detection of H. canis, which has practical applications for the screening of field samples.


Assuntos
Coccidiose/veterinária , Doenças do Cão/diagnóstico , Eucoccidiida/isolamento & purificação , Técnicas de Amplificação de Ácido Nucleico/métodos , Animais , Babesia/genética , Coccidiose/diagnóstico , Primers do DNA , DNA de Protozoário/isolamento & purificação , Doenças do Cão/parasitologia , Cães , Ehrlichia canis/genética , Eucoccidiida/genética , Genoma de Protozoário , Técnicas de Amplificação de Ácido Nucleico/veterinária , RNA Ribossômico 18S/genética , Sensibilidade e Especificidade
19.
Parasitol Res ; 117(12): 3857-3865, 2018 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-30267181

RESUMO

Species of Hepatozoon are parasites frequently recorded in snakes. The species identification of this genus was based mostly on the gametocyte morphology and morphometric calculations. For more reliable results, molecular characterization, an initial step for the correct identification of Hepatozoon species, has been used. This study aimed to determine the prevalence and identification of Hepatozoon species in captive snakes from Brazil. To that end, morphological, morphometric, and molecular data were obtained. A total of 157 snakes; 128 venomous (Crotalus durissus) and 29 non-venomous (Epicrates crassus and Boa constrictor) were screened for Hepatozoon blood parasites. Using light microscopy, 20 (12.78%) snakes were found positive for Hepatozoon spp., of which 6/29 (20.7%) were non-venomous and 14/128 (10.9%) were venomous; all with low parasitemia. Polymerase chain reaction (PCR), performed with the primers HepF300/Hep900, confirmed all 20 (100%) samples positive for hemogregarines. Species of Hepatozoon were identified from eight sequenced samples. Two previously described species, Hepatozoon cuestensis and Hepatozoon musa, were identified. The present study is the first to report H. musa within the snake hosts E. crassus and C. durrisus. In addition, a potentially new Hepatozoon species from B. constrictor was identified.


Assuntos
Coccidiose/veterinária , Crotalus/parasitologia , Eucoccidiida/classificação , Eucoccidiida/genética , Animais , Sequência de Bases , Brasil , Coccidiose/parasitologia , Eucoccidiida/isolamento & purificação , Feminino , Masculino , Parasitemia/parasitologia , Parasitemia/veterinária , Filogenia , Reação em Cadeia da Polimerase
20.
Rev. bras. parasitol. vet ; 27(3): 377-383, July-Sept. 2018. graf
Artigo em Inglês | LILACS | ID: biblio-1042479

RESUMO

Abstract Rangelia vitalii is a haemoparasite that infects erythrocytes, white blood cells and the cytoplasm of endothelial cells of blood capillaries of canids in South America, and has been detected in both domestic dogs and sylvatic canids. Hepatozoon canis is a parasite that infects neutrophils and monocytes of many mammalian hosts. This study reports the infection of Lycalopex gymnocercus from Santa Catarina, Brazil, with R. vitalii and H. canis. The piroplasm was observed on both blood smears and molecular tests. Many large piroplasms were detected inside the erythrocytes, with round, oval, or teardrop-shaped organism, that occurred singly or in pairs. They had an abundant, pale blue cytoplasm and decentral dark red small nucleus. The animal was also infected with H. canis that was detected only by molecular tests. The majority of haematological and biochemistry parameters were within the reference values for domestic dog and wild canids.


Resumo Rangelia vitalii é um hemoparasita que infecta eritrócitos, macrófagos e células endoteliais de canídeos na América do Sul, e vem sendo detectado tanto em cães domésticos quanto em canídeos silvestres. Hepatozoon canis é um parasita que infecta monócitos e neutrófilos de mamíferos. No presente estudo, é descrita a infecção de Lycalopex gymnocercus, proveniente de Santa Catarina, Brasil, por R. vitalii e H. canis. O piroplasma foi diagnosticado nos esfregaços sanguíneos e por técnicas moleculares. Nos eritrócitos foram observados vários merozoítos grandes, ovais, arredondados ou em forma de gota, ocorrendo isoladamente ou em pares. Estes piroplasmas apresentavam citoplasma abundante, corado em azul claro, com núcleo pequeno, avermelhado e descentralizado. O animal apresentou coinfecção com H. canis, que foi diagnosticado somente pelos testes moleculares. A maior parte dos parâmetros hematológicos e bioquímicos do animal estava dentro dos valores de referência para cães domésticos e canídeos silvestres.


Assuntos
Animais , Infecções Protozoárias em Animais/parasitologia , Piroplasmida/isolamento & purificação , Eucoccidiida/isolamento & purificação , Coccidiose/veterinária , Raposas/parasitologia , Filogenia , Infecções Protozoárias em Animais/diagnóstico , Brasil , Piroplasmida/classificação , Piroplasmida/genética , Eucoccidiida/classificação , Eucoccidiida/genética , Coccidiose/diagnóstico , Coccidiose/parasitologia , Coinfecção
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