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1.
Rev Argent Microbiol ; 52(2): 107-114, 2020.
Article in Spanish | MEDLINE | ID: mdl-31787448

ABSTRACT

Bovine abortion causes considerable economic losses to the livestock industry worldwide and is of concern for public health and food safety, given that many abortigenic infectious agents of cattle are zoonotic. Despite its importance, the etiological diagnosis of abortion in cattle is challenging both for veterinary practitioners and laboratory technicians, partly due to the difficulty in recovering aborted fetuses under extensive field conditions for pathological and microbiological diagnostic investigation, and in the early identification of aborted dams. Neospora caninum is a cosmopolitan protozoon identified as one of the main abortigenic agents in cattle worldwide. In this study we propose a comparative seroepidemiological approach for the diagnosis of abortion by N. caninum in dairy cattle. Samples from 12 to 93 cows/heifers with and without recent history of abortion (cases and controls) in four commercial dairy farms were tested. The ratio of controls to cases tested varied from 1:1 to 4.6:1. All samples (n=230) were analyzed by three commercial ELISA kits for the detection of anti-N. caninum antibodies. In all four dairy farms, the proportion of seropositive cows and/or heifers per kit was significantly higher in the cases than in the controls (Odds Ratios=5.13 to 36, p=0.0002 to 0.0485). The agreement among the three kits varied from weak to strong (Cohens kappa coefficients=0.58 to 0.83). We conclude that, despite the imperfect agreement between these kits, all of them allowed to arrive at similar conclusions regarding the statistical association between N. caninum seropositivity and abortion, thus representing a useful tool for the diagnostic approach at the population level under field conditions.


Subject(s)
Abortion, Veterinary/blood , Abortion, Veterinary/parasitology , Antibodies, Protozoan/blood , Cattle Diseases/blood , Cattle Diseases/parasitology , Enzyme-Linked Immunosorbent Assay , Neospora/immunology , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/diagnosis , Animals , Cattle , Cattle Diseases/diagnosis , Dairying , Female , Uruguay
2.
Ticks Tick Borne Dis ; 10(2): 321-327, 2019 02.
Article in English | MEDLINE | ID: mdl-30473318

ABSTRACT

Canine piroplasmoses, caused by Babesia spp., Theileria spp. and Rangelia vitalii, are emerging vector-borne diseases with a worldwide distribution, transmitted by ticks. The aim of this study was to determine the prevalence and perform molecular characterization of piroplasmids in domestic dogs from Asunción city, Paraguay. Blood samples were taken from 384 domestic dogs from Asunción city, Paraguay. DNA was purified from dog blood samples and submitted to nested PCR assays for piroplasmids (18S rRNA) and sequenced for identification and phylogenetic analysis. Overall piroplasmid prevalence in dogs from Paraguay was 6% (23/384 [CI 95% = 3.6-8.4%]). Phylogenetic studies showed that Babesia vogeli was the most prevalent species (91% [21/23]), followed by Theileria equi (4% [1/23]) and Rangelia sp. closely related to R. vitalii (4% [1/23]). Babesia vogeli, T. equi and Rangelia sp. circulate among domestic dogs from Asunción city, and are described for the first time in Paraguay.


Subject(s)
Dogs/parasitology , Piroplasmida/genetics , Protozoan Infections, Animal/epidemiology , Ticks/parasitology , Animals , Animals, Domestic/parasitology , Babesia/genetics , Babesiosis/blood , Babesiosis/epidemiology , DNA, Protozoan/genetics , Paraguay/epidemiology , Phylogeny , Piroplasmida/isolation & purification , Polymerase Chain Reaction , Prevalence , Protozoan Infections, Animal/blood , RNA, Ribosomal, 18S/genetics , Theileria/genetics , Theileriasis/blood , Theileriasis/epidemiology
3.
R. bras. Parasitol. Vet. ; 27(3): 377-383, jul.-set. 2018. ilus
Article in English | VETINDEX | ID: vti-735133

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.(AU)


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.(AU)


Subject(s)
Animals , Foxes/parasitology , Piroplasmia/pathogenicity , Eucoccidiida/pathogenicity , Protozoan Infections, Animal/blood , Coinfection/veterinary
4.
Parasit Vectors ; 11(1): 276, 2018 05 02.
Article in English | MEDLINE | ID: mdl-29716645

ABSTRACT

BACKGROUND: Wetlands are ecosystems in which vectors of avian haemosporidians live and reproduce and where waterbirds join to breed in colonies. Brazil has wetlands at different latitudes, which enables testing the influence of the ecological factors on the prevalence and diversity of haemosporidians. We identified avian haemosporidians in waterbird species in three wetlands and investigated the effects of vector habitat suitability, landscape and host characteristics on the diversity and prevalence of these parasites. METHODS: We created a map with the probability of occurrence of avian haemosporidian vectors using maximum-entropy modelling based on references addressing species known to be vectors of haemosporidians in birds in Brazil. We determined the prevalence and diversity index of haemosporidians in the great egret (Ardea alba) (n = 129) and roseate spoonbill (Platalea ajaja) (n = 180) and compared the findings to data for the wood stork (Mycteria americana) (n = 199). RESULTS: We report the first record of Plasmodium in the family Threskiornithidae: four lineages in the roseate spoonbill, which also presented one lineage of Haemoproteus. In the family Ardeidae, we found three Plasmodium lineages in the great egret. The similar habitat suitability for vectors found in three wetlands explains the pattern of haemosporidian diversity determined for great egret and wood stork populations. Comparisons of haemosporidian diversity within each waterbird species and between regions showed a higher level in the central-western roseate spoonbill population than in the northern population (P = 0.021). Removing the host effect, we discussed the results obtained in terms of characteristics of the Pantanal region. Comparisons of Plasmodium spp. prevalence among waterbird species within the same wetland showed higher level in roseate spoonbill (74%) than those found in the great egret (21%) and wood stork (11%). Excluding the environmental effect, we interpreted result focusing host characteristics that favour infection: time required for nestlings to be covered by feathers and migratory behaviour. CONCLUSIONS: The map of habitat suitability showed that wetlands located in a 30° latitudinal range offer similar conditions for avian vectors species and diversity of haemosporidians. The lineages described in waterbirds were previously identified in birds of prey as Plasmodium paranucleophilum.


Subject(s)
Birds/parasitology , Environment , Haemosporida/physiology , Protozoan Infections, Animal/epidemiology , Wetlands , Animals , Bird Diseases/blood , Bird Diseases/parasitology , Brazil/epidemiology , Disease Vectors , Ecosystem , Genetic Variation , Haemosporida/genetics , Plasmodium/physiology , Prevalence , Protozoan Infections, Animal/blood , Sequence Analysis, DNA
5.
Parasit Vectors ; 10(1): 470, 2017 Oct 10.
Article in English | MEDLINE | ID: mdl-29017602

ABSTRACT

BACKGROUND: Species of Schellackia Reichenow, 1919 have been described from the blood of reptiles distributed worldwide. Recently, Schellackia spp. detected in European and Asian lizards have been molecularly characterised. However, parasites detected in American lizard hosts remain uncharacterised. Thus, phylogenetic affinities between the Old and New World parasite species are unknown. METHODS: In the present study, we characterised morphologically and molecularly the hemococcidian parasites (sporozoites) that infect three lizard hosts from North America and two from South America. RESULTS: In total, we generated 12 new 18S rRNA gene sequences of hemococcidian parasites infecting New World lizard hosts. By the microscopic examination of the smears we identified Schellackia golvani Rogier & Landau, 1975 (ex Anolis carolinensis Voigt) and Schellackia occidentalis Bonorris & Ball, 1955 (ex Uta stansburiana Baird & Girard and Sceloporus occidentalis Baird & Girard) in some samples, but the phylogenetic analysis indicated that all 18S rDNA sequences are distant from Schellackia species found in Old World lizards. In fact, the hemococcidian parasites detected in the New World lizards (including S. occidentalis and S. golvani) were closely related to the genus Lankesterella Labbé, 1899. Consequently, we suggest these two species to be included within the genus Lankesterella. CONCLUSIONS: Life history traits of hemococcidian parasites such as the type of host blood cells infected, host species or number of refractile bodies are not valid diagnostic characteristics to differentiate the parasites between the genera Schellackia and Lankesterella. Indeed, lankesterellid parasites with a different number of refractile bodies had a close phylogenetic origin. Based on the phylogenetic results we provide a systematic revision of the North American hemococcidians. Our recommendation is to include the species formerly described in the genus Schellackia that infect American lizards into Lankesterella (Lankesterellidae) as Lankesterella golvani (Rogier & Landau, 1975) n. comb and L. occidentalis (Bonorris & Ball, 1955) n. comb.


Subject(s)
Apicomplexa/classification , Apicomplexa/genetics , Lizards/parasitology , Phylogeny , Protozoan Infections, Animal/epidemiology , Animals , DNA, Ribosomal/genetics , Eucoccidiida/parasitology , Host Specificity , North America/epidemiology , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/parasitology , RNA, Ribosomal, 18S/genetics , Sequence Analysis, DNA , South America/epidemiology , United States/epidemiology
6.
J Parasitol ; 103(4): 366-376, 2017 08.
Article in English | MEDLINE | ID: mdl-28395577

ABSTRACT

Apicomplexan parasites are obligate parasites of many species of vertebrates. To date, there is very limited understanding of these parasites in the most-diverse group of vertebrates, actinopterygian fishes. While DNA barcoding targeting the eukaryotic 18S small subunit rRNA gene sequence has been useful in identifying apicomplexans in tetrapods, identification of apicomplexans infecting fishes has relied solely on morphological identification by microscopy. In this study, a DNA barcoding method was developed that targets the 18S rRNA gene primers for identifying apicomplexans parasitizing certain actinopterygian fishes. A lead primer set was selected showing no cross-reactivity to the overwhelming abundant host DNA and successfully confirmed 37 of the 41 (90.2%) microscopically verified parasitized fish blood samples analyzed in this study. Furthermore, this DNA barcoding method identified 4 additional samples that screened negative for parasitemia, suggesting this molecular method may provide improved sensitivity over morphological characterization by microscopy. In addition, this PCR screening method for fish apicomplexans, using Whatman FTA preserved DNA, was tested in efforts leading to a more simplified field collection, transport, and sample storage method as well as a streamlining sample processing important for DNA barcoding of large sample sets.


Subject(s)
Apicomplexa/classification , DNA Barcoding, Taxonomic , Fish Diseases/parasitology , Parasitemia/veterinary , Protozoan Infections, Animal/parasitology , Animals , Apicomplexa/genetics , Bayes Theorem , Coral Reefs , DNA Barcoding, Taxonomic/veterinary , DNA, Protozoan/blood , DNA, Protozoan/chemistry , DNA, Ribosomal/chemistry , Fish Diseases/blood , Fish Diseases/epidemiology , Fishes , Likelihood Functions , Parasitemia/epidemiology , Parasitemia/parasitology , Phylogeny , Polymerase Chain Reaction/veterinary , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/epidemiology , RNA, Protozoan/genetics , RNA, Ribosomal, 18S/genetics , Sequence Alignment , United States Virgin Islands/epidemiology
7.
Vet Parasitol ; 217: 113-7, 2016 Feb 15.
Article in English | MEDLINE | ID: mdl-26827871

ABSTRACT

The aim of this study was to develop and validate a SYBR Green qPCR assay to detect and quantify a fragment of the 18S rRNA gene of Rangelia vitalii in canine blood. Repeatability of the qPCR was determined by the intra- and inter-assay variations. The qPCR showed efficiency of E=101.30 (r(2)=0.996), detecting as few as one copy of plasmid containing the target DNA. Specificity of the assay was performed using DNA samples of Babesia canis, B. gibsoni, Ehrlichia canis, E. ewingii and Leishmania sp. No cross-reactivity was observed. Field samples consisting of blood from 265 dogs from Porto Alegre, Brazil were also tested. A total of 24 (9.05%) samples were positive for R. vitalii. Amplicons of 50% of positive samples were confirmed to be R. vitalii by Sanger sequencing. The positive samples had an average of 3.5×10(5) organisms/mL of blood (range: 1.27×10(3)-1.88×10(6)) based on the plasmid-generated standard curve. In conclusion, the SYBR Green qPCR assay developed herein is sensitive and specific and can be used as a diagnostic tool for detection and quantification of R. vitalii in canine blood samples.


Subject(s)
Dog Diseases/diagnosis , Piroplasmida/genetics , Protozoan Infections, Animal/diagnosis , Real-Time Polymerase Chain Reaction/veterinary , Veterinary Medicine/methods , Animals , Brazil , Dog Diseases/blood , Dog Diseases/parasitology , Dogs , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/parasitology , RNA, Ribosomal, 18S/genetics , Real-Time Polymerase Chain Reaction/standards , Sensitivity and Specificity
8.
Int J Parasitol ; 46(1): 41-9, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26348660

ABSTRACT

Generalist pathogens are capable of infecting a wide range of host species, and may pose serious disease emergence threats if accidentally moved outside their native areas. To date little effort has been devoted to identifying geographic areas that may act as reservoirs of generalist pathogens. According to current theory, where host diversity is high, parasite specialisation in one host species may be penalised by reduced host availability, while generalist parasites may benefit from the exploitation of various host species. Therefore natural selection could favor generalist parasites where host diversity is high. Here we explored if, in a highly diverse bird community in Ecuador, a generalist strategy is promoted among local Haemoproteus and Plasmodium blood-borne parasites compared with similar parasite communities throughout the world. We reconstructed the phylogenetic relationships of every parasite lineage in order to understand the evolution of host specificity in this megadiverse area. We found high levels of host generalisation for both parasite genera, and the mean host range of the Haemoproteus community in Ecuador was significantly higher than other parasite communities in other areas outside the Neotropics. Generalist Haemoproteus parasites in this bird community had diverse phylogenetic ancestry, were closely related to specialist parasites and were apparently endemic to the Amazon, showing that different parasites have independently evolved into host generalists in this region. Finally we show that Haemoproteus communities in Ecuador and South America are more generalist than in temperate areas, making this continent a hotspot of generalist Haemoproteus parasites for wild birds.


Subject(s)
Bird Diseases/epidemiology , Bird Diseases/parasitology , Birds/parasitology , Disease Reservoirs/veterinary , Haemosporida/isolation & purification , Protozoan Infections, Animal/epidemiology , Animals , Biodiversity , Biological Evolution , Disease Reservoirs/parasitology , Ecuador , Endemic Diseases , Genetic Variation , Haemosporida/classification , Haemosporida/genetics , Host Specificity , Plasmodium/classification , Plasmodium/genetics , Plasmodium/isolation & purification , Polymerase Chain Reaction , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/parasitology
9.
Parasitol Res ; 115(1): 249-54, 2016 Jan.
Article in English | MEDLINE | ID: mdl-26358101

ABSTRACT

Captive terrestrial tortoises of the species Chelonoidis carbonaria (n = 17) and Chelonoidis denticulata (n = 37) in the state of Minas Gerais, southeastern Brazil, were examined for hematozoans by using a combination of microscopic and molecular methods. Microscopic examination revealed young intra-erythrocytic forms in blood smears from both species of tortoises. The results of PCR, sequencing, and phylogenetic analysis indicated that these parasites belonged to the Haemoproteus spp., whose observed prevalence was 17.6 % in C. carbonaria and 13.5 % in C. denticulata. Phylogenetic analysis indicated that these sequences formed a clade that was grouped with other sequences of Haemoproteus spp. parasites in birds, separate from the clade formed by Haemoproteus spp. of reptiles. This study expands the information regarding the occurrence and distribution of hemosporidia in turtles and is the first study of blood parasites in C. carbonaria.


Subject(s)
Haemosporida/classification , Phylogeny , Protozoan Infections, Animal/epidemiology , Turtles/parasitology , Animals , Base Sequence , Bayes Theorem , Brazil/epidemiology , DNA, Protozoan/blood , DNA, Protozoan/chemistry , DNA, Protozoan/isolation & purification , Female , Haemosporida/genetics , Male , Prevalence , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/parasitology , Sequence Analysis, DNA/veterinary
10.
Exp Parasitol ; 157: 163-9, 2015 Oct.
Article in English | MEDLINE | ID: mdl-26297677

ABSTRACT

The evolutionary origin of wildlife and human malaria parasites (Plasmodium spp.) has been discussed for several decades. The lack of genomic data about species of wildlife haemosporidian parasites related to Plasmodium limits the number of taxa available for phylogenetic analysis. Genomic data about avian parasites of the genus Haemoproteus parasites, the sister genus to Plasmodium are still not available, mainly due to difficulties in obtaining pure DNA of parasites inhabiting nucleated avian host cells. Recent studies show that microgametes of Haemoproteus (Parahaemoproteus) spp. develop in vitro and can be isolated by simple centrifugation, allowing the isolation of pure parasite DNA for genomic studies. However, in vitro development of Haemoproteus (Haemoproteus) spp. has not been investigated, and it is unclear if microgametes of these parasites also can be obtained under in vitro conditions. Here, we provide the first data about the in vitro development of Haemoproteus (Haemoproteus) columbae, a widespread avian haemosporidian parasite, which is specific to pigeons and doves (Columbiformes) and is transmitted by hippoboscid flies (Diptera, Hippoboscidae). In vitro gametogenesis and ookinete development of H. columbae were studied using a strain isolated from a feral Rock Pigeon (Columba livia) in Bogotá-Colombia. The morphological events leading to exflagellation, fertilization and ookinete formation, as well as the rate of development of these stages were followed in vitro at 40 °C, 19 °C and 15 °C for 48 h. Macrogametes, microgametes, zygotes and initial stages of ookinete development were observed in all temperatures, but mature ookinetes were seen only at 40 °C. The largest diversity of sporogonic stages of H. columbae were present at 40 °C however, exflagellation, fertilization of macrogametes and development of immature ookinetes were also observed at 15 °C and 19 °C. Morphological and morphometric features of these stages in vitro were described and illustrated. This study demonstrates a requirement of high temperature for the successful development of mature ookinetes of H. columbae, but not gametes. We show that 1) parasites of the H. (Haemoproteus) subgenus exflagellate in vitro at 15-19 °C, as is the case in H. (Parahaemoproteus) spp. and 2) in vitro exflagellation can be used to obtain pure DNA for genomic studies.


Subject(s)
Bird Diseases/parasitology , Columbidae/parasitology , Haemosporida/growth & development , Protozoan Infections, Animal/parasitology , Animals , Bird Diseases/blood , Bird Diseases/epidemiology , Colombia/epidemiology , Haemosporida/genetics , Molecular Sequence Data , Prevalence , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/epidemiology
11.
Rev Bras Parasitol Vet ; 24(2): 247-50, 2015.
Article in English | MEDLINE | ID: mdl-26154969

ABSTRACT

The aim of this study was to investigate the occurrence of Hepatozoon species infecting dogs in the municipality of Campo Grande, Mato Grosso do Sul (MS), Brazil, using blood samples (n = 165) drawn from dogs. The species Hepatozoon canis was identified in 3.63% of the tested animals using molecular tools. Further studies are needed to determine the clinical relevance of this infection and the main arthropod vectors involved in its transmission.


Subject(s)
Apicomplexa/isolation & purification , Dog Diseases/parasitology , Protozoan Infections, Animal/parasitology , Animals , Apicomplexa/genetics , Brazil , DNA, Protozoan/blood , Dog Diseases/blood , Dog Diseases/diagnosis , Dogs , Molecular Diagnostic Techniques , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/diagnosis
12.
Rev Bras Parasitol Vet ; 24(2): 148-54, 2015.
Article in English | MEDLINE | ID: mdl-26083692

ABSTRACT

Neosporosis is a disease caused by the protozoon Neospora caninum that leads to significant economic losses in many countries. In the present study, we report on use of the recombinant protein NcSRS2 of N. caninum expressed in Pichia pastoris in an indirect immunoenzymatic assay (ELISA) for diagnosing neosporosis infection in sheep and dogs. We observed that the ELISA test yielded specificity of 94.5% and sensitivity of 100% for sheep and specificity of 93.3% and sensitivity of 100% for dogs. We observed that the sensitivity was higher than shown by the indirect fluorescent antibody test, and this was confirmed by means of Western blot. The results from this study suggest that the recombinant protein expressed in P. pastoris is a suitable antigen for use in immunodiagnosis to detect N. caninum in two important species exposed to this parasitosis.


Subject(s)
Antigens, Protozoan/blood , Antigens, Surface/blood , Dog Diseases/blood , Dog Diseases/parasitology , Enzyme-Linked Immunosorbent Assay , Neospora , Protozoan Infections, Animal/blood , Protozoan Proteins/blood , Sheep Diseases/blood , Sheep Diseases/parasitology , Animals , Antigens, Protozoan/biosynthesis , Antigens, Protozoan/immunology , Antigens, Surface/biosynthesis , Antigens, Surface/immunology , Dog Diseases/diagnosis , Dogs , Neospora/immunology , Pichia/metabolism , Protozoan Infections, Animal/diagnosis , Protozoan Proteins/biosynthesis , Protozoan Proteins/immunology , Sheep , Sheep Diseases/diagnosis
13.
Rev. bras. parasitol. vet ; 24(2): 148-154, Apr-Jun/2015. graf
Article in English | LILACS | ID: lil-750757

ABSTRACT

Neosporosis is a disease caused by the protozoon Neospora caninum that leads to significant economic losses in many countries. In the present study, we report on use of the recombinant protein NcSRS2 of N. caninum expressed in Pichia pastoris in an indirect immunoenzymatic assay (ELISA) for diagnosing neosporosis infection in sheep and dogs. We observed that the ELISA test yielded specificity of 94.5% and sensitivity of 100% for sheep and specificity of 93.3% and sensitivity of 100% for dogs. We observed that the sensitivity was higher than shown by the indirect fluorescent antibody test, and this was confirmed by means of Western blot. The results from this study suggest that the recombinant protein expressed in P. pastoris is a suitable antigen for use in immunodiagnosis to detect N. caninum in two important species exposed to this parasitosis.


A neosporose é uma doença causada pelo protozoário Neospora caninum que leva a perdas econômicas importantes em muitos países. No presente estudo, é descrita a utilização da proteína recombinante NcSRS2 de N. caninum expressa em Pichia pastoris em um ensaio imunoenzimático indireto (ELISA) para o diagnóstico de infecção por Neospora em ovelhas e cães. Observou-se, que utilizando-se um ELISA, o teste produziu uma especificidade de 94,5% e uma sensibilidade de 100% para ovinos; e uma especificidade de 93,3% e sensibilidade de 100% para cães. Uma maior sensibilidade foi observada em relação à IFI que foi confirmada por Western blot. Os resultados deste estudo sugerem que a proteína recombinante expressa em P. pastoris é bom antígeno para ser utilizado no diagnóstico imunológico para detectar N. caninum em duas espécies importantes expostas a esta parasitose.


Subject(s)
Animals , Dogs , Protozoan Infections, Animal/blood , Sheep Diseases/blood , Enzyme-Linked Immunosorbent Assay , Protozoan Proteins/blood , Neospora/immunology , Dog Diseases/parasitology , Dog Diseases/blood , Antigens, Protozoan/blood , Antigens, Surface/blood , Pichia/metabolism , Protozoan Infections, Animal/diagnosis , Sheep Diseases/diagnosis , Sheep Diseases/parasitology , Sheep , Protozoan Proteins/biosynthesis , Protozoan Proteins/immunology , Dog Diseases/diagnosis , Antigens, Protozoan/biosynthesis , Antigens, Protozoan/immunology , Antigens, Surface/biosynthesis , Antigens, Surface/immunology
14.
Res Vet Sci ; 97(2): 325-8, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25039064

ABSTRACT

The objective of this survey was to investigate the prevalence of Hepatozoon infection in dogs in the rural and urban areas of Uberlândia, Brazil by PCR and molecular characterization. DNA was obtained from blood samples collected from 346 local dogs from both genders and various ages. Seventeen PCR products from positive blood samples of urban dogs and 13 from the rural dogs were sequenced. Partial sequences of the 18S rRNA gene indicated that all 30 dogs were infected with Hepatozoon canis similar in sequence to H. canis from southern Europe. Four local dog sequences were submitted to GenBank (accessions JN835188; KF692038; KF692039; KF692040). This study indicates that H. canis is the cause of canine hepatozoonosis in Uberlândia and that infection is similarly widespread in rural and urban dogs.


Subject(s)
Apicomplexa/genetics , DNA, Protozoan/genetics , Dog Diseases/epidemiology , Protozoan Infections, Animal/epidemiology , Rural Health , Urban Health , Age Factors , Animals , Brazil/epidemiology , DNA, Protozoan/blood , Databases, Nucleic Acid , Dog Diseases/blood , Dog Diseases/diagnosis , Dogs , Female , Male , Polymerase Chain Reaction/veterinary , Prevalence , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/diagnosis , Sex Factors
15.
Ticks Tick Borne Dis ; 4(4): 341-5, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23567028

ABSTRACT

Hemoparasites were surveyed in 60 free-living pampas deer Ozotoceros bezoarticus from the central area of the Pantanal, known as Nhecolândia, State of Mato Grosso do Sul, Brazil, through the analysis of nested PCR assays and nucleotide sequencing. Blood samples were tested for Babesia/Theileria, Anaplasma spp., and Trypanosoma spp. using nPCR assays and sequencing of the 18S rRNA, msp4, ITS, and cathepsin L genes. The identity of each sequence was confirmed by comparison with sequences from GenBank using BLAST software. Forty-six (77%) pampas deer were positive for at least one hemoparasite, according to PCR assays. Co-infection occurred in 13 (22%) animals. Based on the sequencing results, 29 (48%) tested positive for A. marginale. Babesia/Theileria were detected in 23 (38%) samples, and according to the sequencing results 52% (12/23) of the samples were similar to T. cervi, 13% (3/23) were similar to Babesia bovis, and 9% (2/23) were similar to B. bigemina. No samples were amplified with the primers for T. vivax, while 11 (18%) were amplified with the ITS primers for T. evansi. The results showed pampas deer to be co-infected with several hemoparasites, including species that may cause serious disease in cattle. Pampas deer is an endangered species in Brazil, and the consequences of these infections to their health are poorly understood.


Subject(s)
DNA, Protozoan/genetics , Deer , Protozoan Infections, Animal/blood , Animals , Brazil/epidemiology , Female , Male , Protozoan Infections, Animal/epidemiology , Protozoan Infections, Animal/parasitology
16.
J Parasitol ; 99(3): 522-7, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23259406

ABSTRACT

This study documents the prevalences and lineages of hemoparasites in wood stork nestlings from 3 regions of the American continent: southeastern United States (n = 90), northern Brazil (n = 74), and central-western Brazil (n = 125). Identification was based on PCR amplification of a mitochondrial small subunit ribosomal RNA gene. A fragment of the hemoparasite cytochrome B gene in infected individuals was utilized for Bayesian phylogenetic analysis. Four wood stork nestlings were infected by Haemoproteus, 1 from northern Brazil and 3 from the United States, and all shared the same haplotype. Morphological analysis confirmed the infection of the U.S. birds by Haemoproteus. Infection by Plasmodium was found in wood stork nestlings from northern (6) and central-western Brazil (14). Five Plasmodium lineages (MYCAMP1-2, and MYCAMP4-6) were found in the Brazilian central-western region and 3 Plasmodium lineages (MYCAMP2-3, and MYCAMP7) were found in the northern region. The most prevalent haplotype (MYCAMP2) differs from the others by 1 mutation, and the less prevalent haplotypes are derived from MYCAMP2. We did not find Plasmodium or Haemoproteus in nestlings younger than 15 and 30 days old, respectively. This is the first documentation of Plasmodium and Haemoproteus infection in wood storks in Brazilian breeding populations. Potential connectivity among wood stork populations was indirectly supported by the presence of identical Haemoproteus lineages in U.S. and northern Brazilian populations, and by the presence of identical Plasmodium haplotypes in the northern and central-western Brazilian populations.


Subject(s)
Bird Diseases/parasitology , Haemosporida/isolation & purification , Malaria, Avian/parasitology , Protozoan Infections, Animal/parasitology , Animals , Bayes Theorem , Bird Diseases/blood , Bird Diseases/epidemiology , Birds , Brazil/epidemiology , DNA, Protozoan/blood , DNA, Protozoan/isolation & purification , Florida/epidemiology , Georgia/epidemiology , Haemosporida/classification , Haemosporida/genetics , Malaria, Avian/blood , Malaria, Avian/epidemiology , Plasmodium/classification , Plasmodium/genetics , Plasmodium/isolation & purification , Polymerase Chain Reaction/veterinary , Prevalence , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/epidemiology , Wetlands
17.
J Parasitol ; 99(3): 513-21, 2013 Jun.
Article in English | MEDLINE | ID: mdl-23240808

ABSTRACT

Here we describe Haemoproteus (Haemoproteus) multivolutinus n. sp. from a tambourine dove (Turtur timpanistria) of Uganda and Haemoproteus (Haemoproteus) paramultipigmentatus n. sp. (Haemosporida, Haemoproteidae) from the Socorro common ground dove (Columbina passerina socorroensis) of Socorro Island, Mexico. These parasites are described based on the morphology of their blood stages and segments of the mitochondrial cytochrome b gene that can be used for molecular identification and diagnosis of these species. Gametocytes of H. multivolutinus possess rod-like pigment granules and are evenly packed with volutin, which masks pigment granules and darkly stains both macro- and microgametocytes in the early stages of their development. Based on these 2 characters, H. multivolutinus can be readily distinguished from other species of hemoproteids parasitizing columbiform (Columbiformes) birds. Haemoproteus paramultipigmentatus resembles Haemoproteus multipigmentatus; it can be distinguished from the latter parasite primarily due to the broadly ovoid shape of its young gametocytes and significantly fewer pigment granules in its fully developed gametocytes. We provide illustrations of blood stages of the new species, and phylogenetic analyses identify DNA lineages closely related to these parasites. Cytochrome b lineages of Haemoproteus multivolutinus and H. paramultipigmentatus cluster with hippoboscid-transmitted lineages of hemoproteids; thus these parasites likely belong to the subgenus Haemoproteus. We emphasize the importance of using cytochrome b sequences in conjunction with thorough microscopic descriptions to facilitate future identification of these and other avian hemosporidian species.


Subject(s)
Bird Diseases/parasitology , Columbidae/parasitology , Haemosporida/classification , Protozoan Infections, Animal/parasitology , Animals , Bird Diseases/blood , Bird Diseases/epidemiology , Cytochromes b/genetics , Erythrocytes/parasitology , Haemosporida/genetics , Haemosporida/ultrastructure , Mexico/epidemiology , Molecular Sequence Data , Phylogeny , Prevalence , Protozoan Infections, Animal/blood , Protozoan Infections, Animal/epidemiology , Uganda/epidemiology
18.
Rev Bras Parasitol Vet ; 21(3): 243-8, 2012.
Article in English | MEDLINE | ID: mdl-23070434

ABSTRACT

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.


Subject(s)
Alanine Transaminase/blood , Aspartate Aminotransferases/blood , Creatine Kinase/blood , Dog Diseases/blood , Dog Diseases/parasitology , Protozoan Infections, Animal/blood , Acute Disease , Animals , Dog Diseases/enzymology , Dogs , Female , Protozoan Infections, Animal/enzymology
19.
Rev. bras. parasitol. vet ; 21(3): 243-248, July-Sept. 2012. ilus
Article in English | LILACS | ID: lil-653712

ABSTRACT

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.


Rangelia vitalii é um protozoário que causa doença em cães, sendo a anemia o achado laboratorial mais frequente. No entanto, existem poucos estudos sobre as alterações bioquímicas em cães infectados com o protozoário. Assim, este estudo tem como objetivo investigar as alterações bioquímicas de cães experimentalmente infectados com R. vitalii na fase aguda da infecção. Para o estudo, foram utilizados 12 cães fêmeas (com idade entre 6 a 12 meses e peso entre 4 a 7 kg), divididos em dois grupos. O grupo A (n = 5) foi composto de animais saudáveis e o grupo B (n = 7) de animais infectados. Amostras de sangue foram coletadas nos dias zero, dez, vinte e trinta PI, utilizando tubos sem anticoagulante para obtenção de soro e análise dos parâmetros bioquímicos. Foi observado um aumento na alanino aminotransferase (ALT) no dia 20 PI (P < 0,05) e aumento na creatinoquinase (CK) e aspartato aminotransferase (AST) em todo o período experimental (P < 0,05). Não foram observadas alterações séricas na gama-glutamiltransferase, uréia e creatinina. Portanto, é possível concluir que a infecção experimental por R. vitalii causa alterações no perfil bioquímico, com aumento na ALT, CK e AST.


Subject(s)
Animals , Dogs , Female , Alanine Transaminase/blood , Aspartate Aminotransferases/blood , Creatine Kinase/blood , Dog Diseases/blood , Dog Diseases/parasitology , Protozoan Infections, Animal/blood , Acute Disease , Dog Diseases/enzymology , Protozoan Infections, Animal/enzymology
20.
R. bras. Parasitol. Vet. ; 21(3): 243-248, July-Sept. 2012. ilus
Article in English | VETINDEX | ID: vti-12621

ABSTRACT

Rangelia vitalii is a protozoon that causes diseases in dogs, and anemia is the most common laboratory finding. However, few studies on the biochemical changes in dogs infected with this protozoon exist. Thus, this study aimed to investigate the biochemical changes in dogs experimentally infected with R. vitalii, during the acute phase of the infection. For this study, 12 female dogs (aged 6-12 months and weighing between 4 and 7 kg) were used, divided in two groups. Group A was composed of healthy dogs (n = 5); and group B consisted of infected animals (n = 7). Blood samples were collected on days 0, 10, 20 and 30 after infection, using tubes without anticoagulant to obtain serum and analyze the biochemical parameters. An increase in alanine aminotransferase (ALT) on day 20 (P < 0.05) was observed. Also, increased creatine kinase (CK) and aspartate aminotransferase (AST) levels were observed throughout the experimental period (P < 0.05). No changes in the serum gamma-glutamyltransferase, urea and creatinine levels were observed. Thus, is possible to conclude that experimental infection with R. vitalii in dogs causes changes to the biochemical profile, with increased ALT, AST and CK enzyme levels.(AU)


Rangelia vitalii é um protozoário que causa doença em cães, sendo a anemia o achado laboratorial mais frequente. No entanto, existem poucos estudos sobre as alterações bioquímicas em cães infectados com o protozoário. Assim, este estudo tem como objetivo investigar as alterações bioquímicas de cães experimentalmente infectados com R. vitalii na fase aguda da infecção. Para o estudo, foram utilizados 12 cães fêmeas (com idade entre 6 a 12 meses e peso entre 4 a 7 kg), divididos em dois grupos. O grupo A (n = 5) foi composto de animais saudáveis e o grupo B (n = 7) de animais infectados. Amostras de sangue foram coletadas nos dias zero, dez, vinte e trinta PI, utilizando tubos sem anticoagulante para obtenção de soro e análise dos parâmetros bioquímicos. Foi observado um aumento na alanino aminotransferase (ALT) no dia 20 PI (P < 0,05) e aumento na creatinoquinase (CK) e aspartato aminotransferase (AST) em todo o período experimental (P < 0,05). Não foram observadas alterações séricas na gama-glutamiltransferase, uréia e creatinina. Portanto, é possível concluir que a infecção experimental por R. vitalii causa alterações no perfil bioquímico, com aumento na ALT, CK e AST.(AU)


Subject(s)
Animals , Female , Dogs , Alanine Transaminase/blood , Aspartate Aminotransferases/blood , Creatine Kinase/blood , Dog Diseases/blood , Dog Diseases/parasitology , Protozoan Infections, Animal/blood , Acute Disease , Dog Diseases/enzymology , Protozoan Infections, Animal/enzymology
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