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1.
Vet Res ; 44: 1, 2013 Jan 04.
Artículo en Inglés | MEDLINE | ID: mdl-23289625

RESUMEN

The present study was aimed at investigating the effect of experimental infection by Trypanosoma vivax in different stages of pregnancy, determining the pathogenesis of reproductive failure, and confirming transplacental transmission. We used 12 pregnant ewes distributed into four experimental groups: G1, was formed by three ewes infected with T. vivax in the first third of pregnancy (30 days); G2 comprised three infected ewes in the final third of pregnancy (100 days); G3 and G4 were composed of three non-infected ewes with the same gestational period, respectively. Each ewe of G1 and G2 was inoculated with 1.25×10(5) tripomastigotes. Clinical examination, determination of parasitemia, serum biochemistry (albumin, total protein, glucose, cholesterol, and urea), packed cell volume (PCV), serum progesterone, and pathological examination were performed. Placenta, amniotic fluid, blood and tissues from the fetuses and stillbirths were submitted to PCR. Two ewes of G1 (Ewe 1 and 3) presented severe infection and died in the 34th and 35th days post-infection (dpi), respectively; but both fetuses were recovered during necropsy. In G2, Ewe 5 aborted two fetuses on the 130th day (30 dpi) of pregnancy; and Ewe 6 aborted one fetus in the 140th day (40 dpi) of gestation. Ewes 2 and 4 delivered two weak lambs that died five days after birth. Factors possibly involved with the reproductive failure included high parasitemia, fever, low PCV, body score, serum glucose, total protein, cholesterol, and progesterone. Hepatitis, pericarditis, and encephalitis were observed in the aborted fetuses. The presence of T. vivax DNA in the placenta, amniotic fluid, blood, and tissues from the fetuses confirms the transplacental transmission of the parasite. Histological lesion in the fetuses and placenta also suggest the involvement of the parasite in the etiopathogenesis of reproductive failure in ewes.


Asunto(s)
Aborto Veterinario/parasitología , Transmisión Vertical de Enfermedad Infecciosa/veterinaria , Enfermedades de las Ovejas/transmisión , Trypanosoma vivax/fisiología , Tripanosomiasis/veterinaria , Animales , Brasil , Femenino , Parasitemia/parasitología , Parasitemia/transmisión , Parasitemia/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Embarazo , Distribución Aleatoria , Ovinos , Enfermedades de las Ovejas/parasitología , Tripanosomiasis/parasitología , Tripanosomiasis/transmisión
2.
Exp Parasitol ; 128(4): 324-7, 2011 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-21501609

RESUMEN

Tick-borne diseases in horses are caused by the intraerythrocytic protozoan parasites Theileria equi and Babesia caballi. Although T. equi is highly endemic in Latin America, the New World vector of this important parasite is controversial. The aim of this study was to test the ability of nymph Amblyomma cajennense ticks acquire infection by T. equi following feeding on infected horses. Three experiments were performed: tick acquisition of T. equi from an experimentally infected horse, tick acquisition of T. equi from naturally infected foals and tick acquisition of T. equi from a chronically infected horse. A. cajennense adults were dissected and salivary glands were collected in aliquots. Methyl green pyronin staining of the salivary glands did not show the presence of hypertrophy of acini or cell nuclei normally suggestive of Theileria spp. infection. The pools of salivary glands were negative for Theileria DNA in nested PCR assays. Histopathological analysis failed to detect sporoblast and sporozoites of T. equi in salivary gland acini. This study was not able to observe infection of the A. cajennense by T. equi.


Asunto(s)
Vectores Arácnidos/parasitología , Enfermedades de los Caballos/transmisión , Ixodidae/parasitología , Theileria/fisiología , Theileriosis/transmisión , Enfermedad Aguda , Animales , Bovinos , Enfermedad Crónica , ADN Protozoario/aislamiento & purificación , Electroforesis en Gel de Agar/veterinaria , Femenino , Enfermedades de los Caballos/parasitología , Caballos , Masculino , Ninfa/parasitología , Parasitemia/parasitología , Parasitemia/transmisión , Parasitemia/veterinaria , Reacción en Cadena de la Polimerasa/veterinaria , Glándulas Salivales/parasitología , Theileria/aislamiento & purificación
3.
Exp Parasitol ; 122(3): 192-5, 2009 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-19324040

RESUMEN

This paper describes the in vitro multiplication process of Babesia bigemina sporokinetes in a cell line (IDE8) from Ixodes scapularis ticks. The inoculum was obtained from hemolymph of engorged females of Rhipicephalus (Boophilus) microplus ticks naturally infected with B. bigemina. These ticks had been fed on calves living in a tick endemic farm in Brazil. Microscopic morphological details are shown to describe the development of the parasite in the tick cells; the identity of the parasite was confirmed by a duplex PCR method.


Asunto(s)
Babesia/crecimiento & desarrollo , Ixodes/parasitología , Animales , Babesia/genética , Babesia/aislamiento & purificación , Bovinos , Enfermedades de los Bovinos/parasitología , Línea Celular , ADN Protozoario/análisis , Femenino , Hemolinfa/parasitología , Ixodes/citología , Ixodes/embriología , Reacción en Cadena de la Polimerasa , Rhipicephalus/parasitología
4.
Vet Parasitol ; 161(3-4): 270-5, 2009 May 12.
Artículo en Inglés | MEDLINE | ID: mdl-19339119

RESUMEN

Rhipicephalus sanguineus ticks are distributed throughout the world, especially in those areas in which dogs are in close contact with humans. R. sanguineus and fleas are regarded as the main ectoparasites infesting dogs in Brazil. Besides causing direct damage during the blood feeding process, this tick species can also transmit pathogens to dogs and humans. Despite its importance in Brazil, data regarding the seasonality of R. sanguineus are limited, especially with regard to natural infestations of dogs. The present study aimed to evaluate the seasonality of R. sanguineus on dogs living in Belo Horizonte, state of Minas Gerais. From August 2006 to July 2007, ticks were collected monthly from 12 adult dogs in nine houses, which were located in two districts in the north region of the city. In parallel, canine clients of a pet care department of the small animal veterinary clinic were examined for the presence of ticks before bathing and/or clipping. The climatic data recorded for Belo Horizonte during the experimental period were: mean temperature 18.6 degrees C; relative air humidity 56.5%; rainfall 37mm. The only species of ticks identified from all infested dogs was R. sanguineus, which was found in all its development stages. Among dogs living in houses, three tick population peaks were observed (August, February, and June), suggesting the occurrence of three generations per year in Belo Horizonte. A total of 7318 ticks were collected, of which 5422 were adult ticks and 1896 represented immature stages (744 larvae and 1152 nymphs). The monthly inspection of dogs living in houses demonstrated significantly higher parasitism during the dry season (p<0.05). A total of 2848 dogs from the pet care department of the small animal veterinary clinic were examined, of which 222 (7.8%) were infested with ticks and the percentage of infested dogs in the dry season was higher (p<0.05) than in the hot wet. The percentage of male dogs infested with ticks was significantly higher (58.29%) than the percentage of infested female dogs (41.70%). This study of the dynamics of R. sanguineus infestations in Belo Horizonte will contribute to establishing appropriate measures to control tick infestations in dogs.


Asunto(s)
Enfermedades de los Perros/parasitología , Rhipicephalus sanguineus/fisiología , Infestaciones por Garrapatas/veterinaria , Animales , Brasil/epidemiología , Enfermedades de los Perros/epidemiología , Perros , Larva , Ninfa , Dinámica Poblacional , Estaciones del Año , Infestaciones por Garrapatas/epidemiología , Infestaciones por Garrapatas/parasitología
5.
Braz J Microbiol ; 40(2): 399-403, 2009 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-24031379

RESUMEN

A Brazilian isolate of Anaplasma marginale with appendage was successfully established and maintained in vitro in a tick cell line (IDE8). Infection was confirmed by optical and transmission electron microscopy. In addition, primers MSP1aNF2 and MSP1aNR2 amplified products from DNA extracted from infected IDE8 cells. Comparisons with partial sequences of the msp1α gene and the complete genome of A. marginale confirmed that the sequences of amplified fragments were from the A. marginale genome. This is the first establishment of a Brazilian A. marginale isolate in tick cells, representing a new system for biological and molecular studies and also a new source of material for diagnosis and development of vaccines.

6.
Prev Vet Med ; 79(2-4): 155-62, 2007 May 16.
Artículo en Inglés | MEDLINE | ID: mdl-17210194

RESUMEN

Epidemiological aspects of Babesia vogeli infection were studied in the canine population of a rural town located in the Brazilian "Drought Polygon" of the state of Minas Gerais, Brazil. The survey was carried out in March 2003, when 505 dogs were identified and their characteristics registered on appropriate forms. Blood samples were collected at this time and again in June, September and December 2003. Serum samples were tested by the indirect fluorescent antibody test (IFAT) to detect antibodies against B. vogeli. The prevalence of anti-B. vogeli antibodies was 18.8%; however, no correlations were found between prevalence of infection and the age or gender of the animals. Cross-bred dogs presented a higher chance of acquiring infection in comparison to pure-bred dogs. Significant differences concerning the incidence of the disease were found during the period April-June in comparison to other months, demonstrating that transmission of B. vogeli is related to seasonal variations of tick infestations. The results indicate that climatic factors within the semiarid area interfere directly in the epidemiology of canine babesiosis.


Asunto(s)
Anticuerpos Antiprotozoarios/sangre , Babesiosis/veterinaria , Enfermedades de los Perros/epidemiología , Infestaciones por Garrapatas/veterinaria , Factores de Edad , Animales , Vectores Arácnidos/parasitología , Babesia/inmunología , Babesiosis/epidemiología , Brasil/epidemiología , Perros , Femenino , Técnica del Anticuerpo Fluorescente Indirecta/métodos , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Masculino , Estaciones del Año , Estudios Seroepidemiológicos , Factores Sexuales , Infestaciones por Garrapatas/epidemiología , Garrapatas/parasitología
7.
Parasitol Res ; 102(1): 63-8, 2007 Dec.
Artículo en Inglés | MEDLINE | ID: mdl-17828553

RESUMEN

Blood samples were collected from 487 adult horses, including 83 pregnant mares, at a slaughterhouse located in Araguari, Minas Gerais State, Brazil. For each blood sample, the packed cell volume (PCV) was determined, and Giemsa-stained smears were microscopically examined for the presence of hemoparasites. The plasma was examined by the indirect fluorescent antibody test for detection of antibodies against Babesia caballi and Theileria equi. In addition, DNA was extracted and analyzed by a multiplex real-time polymerase chain reaction (PCR), specific for B. caballi and T. equi. Products of PCR were sequenced and compared with each other and with known sequences. The serological results showed a total prevalence of 91.0% for T. equi and 83.0% for B. caballi, while by PCR, prevalences of 59.7% for T. equi and 12.5% for B. caballi were observed. However, no correlations were seen between positivity (neither by serology nor by PCR) and PCV values. As expected, the microscopic examination of blood smears showed low sensitivity in detecting the infections when compared to the PCR. Only 35 out of 570 blood smears were positive, with parasitemias below 0.1%. No congenital transmission was detectable. As far as sequencing is concerned, no differences were seen among the isolates of each species nor among them and known sequences available. These results confirm, by molecular methods, the high prevalence rates of T. equi and B. caballi infections in carrier horses in Brazil. However, no diversity was observed among the isolates within the studied regions.


Asunto(s)
Babesia/clasificación , Babesia/genética , Babesiosis/veterinaria , Enfermedades de los Caballos/parasitología , Theileria/clasificación , Theileria/genética , Theileriosis/parasitología , Animales , Babesiosis/sangre , Babesiosis/epidemiología , Babesiosis/parasitología , Brasil/epidemiología , Femenino , Enfermedades de los Caballos/sangre , Enfermedades de los Caballos/epidemiología , Caballos , Prevalencia , Theileriosis/sangre , Theileriosis/epidemiología
8.
Ticks Tick Borne Dis ; 7(6): 1102-1108, 2016 10.
Artículo en Inglés | MEDLINE | ID: mdl-27612925

RESUMEN

The aim of the study was to isolate and establish an Anaplasma marginale strain from Brazilian brown brocket deer, Mazama gouazoubira, in the Ixodes scapularis cell line IDE8. Blood from a free-living adult female M. gouazoubira naturally infected with A. marginale (MGI5) was inoculated intravenously into a splenectomized calf. When A. marginale rickettsemia was 2.5%, blood was collected and cryopreserved in liquid nitrogen with dimethylsulfoxide (DMSO). IDE8 cell cultures were infected with calf blood inoculated with the A. marginale (MG15) isolate. The cultures were monitored by examination of Giemsa-stained cytocentrifuge smears. Light microscopy of stained IDE8 samples revealed the first inclusions of A. marginale (MGI5) at 48days post-inoculation (d.p.i). The IDE8-infected cells contained parasitophorous vacuoles with amorphous material and a few cocci-like organisms. A sample from IDE8-infected cells from the 16th subculture (336 d.p.i.) was analyzed by nPCR, nucleotide sequencing, electron microscopy, and an indirect fluorescent antibody test (IFAT). The IFAT highlighted some IDE8-infected cells with intense fluorescence in the parasitophorous vacuole, while in other cells, fluorescence was observed only at the periphery. DNA from a culture of the MG15 isolate was amplified with A. marginale msp4 gene primers, and nucleotide sequencing of the PCR product and BLAST software analysis further confirmed 100% identity with the MGI5 blood isolate (GenBank no. JN022558.1). Electron microscopy revealed increased numbers of lysosomes in the cytoplasm of IDE8 cells. Several cells exhibited large vacuoles containing cellular debris and amorphous material. After the 29th subculture, it was not possible to detect compatible Anaplasma structures by light microscopy, and subculture samples tested negative in nPCR. Despite the failure of the attempt to establish A. marginale (MGI5) in IDE8 cells, the results demonstrated the isolate's ability to infect, survive and multiply, although in limited numbers, in IDE8 cells.


Asunto(s)
Anaplasma marginale/aislamiento & purificación , Anaplasmosis/microbiología , Técnicas Bacteriológicas , Ciervos/microbiología , Anaplasma marginale/fisiología , Anaplasmosis/epidemiología , Animales , Brasil/epidemiología , Bovinos , Línea Celular , Técnica del Anticuerpo Fluorescente Indirecta , Ixodes , Microscopía , Reacción en Cadena de la Polimerasa
9.
Ticks Tick Borne Dis ; 7(1): 80-84, 2016 Feb.
Artículo en Inglés | MEDLINE | ID: mdl-26381444

RESUMEN

Bovine anaplasmosis is a disease caused by the intraerythrocytic rickettsia species Anaplasma marginale and results in great economic losses in tropical and subtropical regions. Vertical transmission is an important phenomenon that contributes to the persistence of different strains of the agent within the same herd. The identification of new strains and genetic characterization studies are essential to understanding their epidemiology and virulence and for vaccine development. The aim of this study was to perform molecular and phylogenetic characterizations of a new vertically transmitted strain from A. marginale and to evaluate its virulence by experimental inoculation of rickettsia-free calves. Thirty newborn Holstein calves were subjected to molecular tests for the detection of A. marginale, Babesia bovis and Babesia bigemina. Calves positive for A. marginale (n=3) were splenectomized and monitored for the clinical manifestations of anaplasmosis. Blood samples from one of the calves that presented rickettsemia of 42.8% and spontaneous recovery of clinical parameters were used for molecular and phylogenetic characterization (msp1a gene), and inoculum production was used for the evaluation of virulence. This strain was identified as UFMG3. Three tandem repeat forms (13 and MGI19) were identified from the analysis of the msp1a gene, in which the form MGI19 appeared twice. Analysis of these repeats revealed the presence of the sequences QASTSS and SSASGQQQESS and of aspartic acid (D) at position 20 of both repeats. Phylogenetic analysis showed a close relationship among the UFMG3, MGI19 and UFMG2 strains. For virulence evaluation, six Holstein calves were inoculated intravenously with 2×10(7)A. marginale UFMG3-infected erythrocytes. The calves showed maximum rickettsemia of 5.1%, a moderate decrease in packed cell volume and spontaneous recovery of clinical parameters without the need for treatment. The results of experimental inoculation suggest that the strain A. marginale UFMG3 has low virulence and potential application for use as a live vaccine against A. marginale.


Asunto(s)
Anaplasma marginale/genética , Anaplasmosis/parasitología , Enfermedades de los Bovinos/microbiología , Anaplasma marginale/patogenicidad , Animales , Bovinos , Enfermedades de los Bovinos/epidemiología , Femenino , Variación Genética , Transmisión Vertical de Enfermedad Infecciosa , Masculino , Filogenia , Embarazo , Esplenectomía , Virulencia
10.
Ticks Tick Borne Dis ; 7(5): 1017-1023, 2016 07.
Artículo en Inglés | MEDLINE | ID: mdl-27344507

RESUMEN

Approximately 50% of buffalo herds in Brazil are located in Pará state in northern Brazil. There are several properties where cattle and buffalo live and graze together, and thus, buffalo pathogens may threaten the health of cattle and vice versa. Therefore, knowledge of infectious agents of buffalo is essential for maintaining healthy livestock. Clinical disease caused by Theileria and Babesia parasites in the Asian water buffalo is not common, although these animals may act as reservoir hosts, and the detection of these hemoparasites in buffaloes is as important as it is in cattle. Studies of the infection of buffaloes by hemoparasites in Brazil are scarce. The objective of the present study was to investigate the occurrence of Piroplasmida parasites in Asian water buffaloes in the state of Pará in the Amazon region of Brazil using nested PCR assays and phylogenetic analysis. The 18S rRNA gene and ITS complete region were amplified from DNA extracted from blood samples collected from 308 apparently healthy buffaloes bred on six properties in the state of Pará, Brazil. The prevalence of positive buffalo samples was 4.2% (13/308) for Theileria spp., 3.6% (11/308) for Babesia bovis and 1% (3/308) for Babesia bigemina. Animals infected with Theileria were detected in 50% (3/6) of the assessed properties. Phylogenetic analyses indicated that the Theileria species detected in this study were closely related to Theileria buffeli, Theileria orientalis and Theileria sinensis. To our knowledge, this is the first report of Theileria in Asian water buffaloes in the Americas. The majority of Theileria-positive buffaloes (11/13) belong to a property that has a history of animals presenting lymphoproliferative disease of unknown etiology. Therefore, the present research suggests that this disorder can be associated with Theileria infection in this property. Our results provide new insights on the distribution and biological aspects of hemoparasites transmissible from buffaloes to cattle.


Asunto(s)
Babesia/aislamiento & purificación , Babesiosis/parasitología , Búfalos/parasitología , Enfermedades de los Bovinos/parasitología , Theileria/aislamiento & purificación , Theileriosis/parasitología , Animales , Babesia/clasificación , Babesia/genética , Babesiosis/epidemiología , Babesiosis/transmisión , Brasil/epidemiología , Bovinos , Enfermedades de los Bovinos/epidemiología , Enfermedades de los Bovinos/transmisión , ADN Protozoario/genética , Variación Genética , Trastornos Linfoproliferativos/etiología , Trastornos Linfoproliferativos/parasitología , Trastornos Linfoproliferativos/veterinaria , Filogenia , Reacción en Cadena de la Polimerasa , Theileria/clasificación , Theileria/genética , Theileriosis/epidemiología , Theileriosis/transmisión
11.
Ticks Tick Borne Dis ; 6(3): 242-5, 2015 Apr.
Artículo en Inglés | MEDLINE | ID: mdl-25661650

RESUMEN

Information on Anaplasma phagocytophilum in Brazil is very restricted. The aim of this study was to report clinical, parasitological, hematological and molecular evidence of a natural A. phagocytophilum infection of an urban Brazilian dog. The dog was an eight-month-old male French bulldog. Veterinary clinical examinations were performed three times: in April, June and December 2013. Biochemical and hematological analyses were performed during all examinations, and blood samples were collected for parasitological surveys in June and December. Morulae were present within neutrophils in blood smears from June. Both samples were PCR positive for A. phagocytophilum and Ehrlichia spp. Phylogenetic analysis revealed that the phylogenetic topology placed samples from this study in close proximity to other A. phagocytophilum isolates. Ehrlichia isolates from this dog were 100% identical to E. canis isolates, thus E. canis and A. phagocytophilum co-infection was diagnosed in this dog. Lethargy and skin lesions were the clinical signs observed in this dog. Abnormal hematological parameters, among those, severe thrombocytopenia, were observed in all three occasions. This finding highlights the growing importance of A. phagocytophilum in South America.


Asunto(s)
Anaplasma phagocytophilum/aislamiento & purificación , Coinfección/veterinaria , Enfermedades de los Perros/diagnóstico , Ehrlichia canis/aislamiento & purificación , Ehrlichiosis/veterinaria , Anaplasma phagocytophilum/genética , Animales , Secuencia de Bases , Brasil , ADN Bacteriano/química , ADN Bacteriano/genética , Perros , Ehrlichia canis/genética , Ehrlichiosis/diagnóstico , Femenino , Masculino , Datos de Secuencia Molecular , Filogenia , Análisis de Secuencia de ADN
12.
Ticks Tick Borne Dis ; 6(4): 522-9, 2015 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-25943785

RESUMEN

Anaplasma marginale is an economically important tick-borne pathogen of cattle that causes bovine anaplasmosis. A wide range of geographic strains of A. marginale have been isolated from cattle, several of which have been characterized using genomics and proteomics. While many of these strains have been propagated in tick lines, comparative analyses after propagation in tick cells have not been reported. The overall purpose of this research therefore was to compare the degree of conservation of selected genes after propagation in tick cell culture among A. marginale strains from the U.S. (the Virginia strain) and Brazil (UFMG1 and UFMG2 strains). The genes studied herein included those which encode the proteins HSP70 and SODB involved in heat shock and stress responses, respectively, and two genes that encode major surface proteins MSP4 and MSP5. Strain identities were first confirmed by sequencing the tandem repeats of the msp1a gene which encodes for the adhesin, MSP1a. The results of these studies demonstrated that the genes encoding for both stress response and heat shock proteins were highly conserved among the three A. marginale strains. Antibodies specific for MSP4, MSP5, SODB and HSP70 proteins were used to further characterize the A. marginale strains, and they reacted with all of these strains propagated in tick cell culture, providing further evidence for antigenic conservation. Although antigenic differences were not found among the three A. marginale strains, multi-locus sequence analysis (MLSA) performed with nucleotide sequences of these genes demonstrated that the A. marginale Brazilian and U.S. strains fall in different clades. These results showed that phylogenetically distant strains of A. marginale are antigenically conserved, even after several in vitro passages, supporting the use of some of the above conserved proteins as candidates for universal vaccines.


Asunto(s)
Anaplasma marginale/aislamiento & purificación , Anaplasmosis/inmunología , Vectores Arácnidos/microbiología , Proteínas Bacterianas/genética , Proteínas Bacterianas/inmunología , Enfermedades de los Bovinos/inmunología , Garrapatas/microbiología , Anaplasma marginale/clasificación , Anaplasma marginale/genética , Anaplasma marginale/crecimiento & desarrollo , Anaplasmosis/microbiología , Animales , Variación Antigénica , Brasil , Bovinos , Enfermedades de los Bovinos/microbiología , Secuencia Conservada , Datos de Secuencia Molecular , Filogenia , Estados Unidos
13.
Vet Parasitol ; 107(1-2): 169-77, 2002 Jul 29.
Artículo en Inglés | MEDLINE | ID: mdl-12072223

RESUMEN

The present study, describes the antigenic characterization of a Brazilian isolate of Anaplasma marginale with appendage (tail). A panel of monoclonal antibodies (McAbs) was produced and tested by the indirect fluorescent antibody test (IFAT), ELISA and Western blotting, and used to characterize two isolates of A. marginale (one with appendage and another without appendage). Among the clones produced, eight recognized antigenic proteins, with molecular weights varying from 18.4 to 66kDa. In Western blotting, the McAb reacted against a 45kDa antigen, which was shown, by the IFAT, to be located in the tail. Immunocytochemistry confirmed the tail specificity of the monoclonal reacting against the 45kDa antigen. The panel of McAb produced has a potential use in discriminating morphologically distinct A. marginale isolates. The present study, demonstrates the occurrence of antigenic diversity among Brazilian isolates of A. marginale.


Asunto(s)
Anaplasma/inmunología , Variación Antigénica , Antígenos Bacterianos/análisis , Anaplasma/clasificación , Anaplasma/ultraestructura , Anaplasmosis/inmunología , Anaplasmosis/microbiología , Animales , Anticuerpos Monoclonales/inmunología , Western Blotting/veterinaria , Brasil , Bovinos , Enfermedades de los Bovinos/inmunología , Enfermedades de los Bovinos/microbiología , Ensayo de Inmunoadsorción Enzimática/veterinaria , Técnica del Anticuerpo Fluorescente Indirecta/veterinaria , Inmunohistoquímica/veterinaria , Microscopía Inmunoelectrónica/veterinaria , Peso Molecular
14.
Vet Parasitol ; 121(3-4): 307-16, 2004 May 26.
Artículo en Inglés | MEDLINE | ID: mdl-15135871

RESUMEN

Anaplasma marginale (Rickettsiales: Anaplasmataceae), a tick-borne pathogen of cattle, is endemic in tropical and subtropical regions of the world, and many isolates of A. marginale may occur in a given geographic area. Phylogenetic relationships have been reported for A. marginale isolates from the US using gene and protein sequences of MSP1a and msp4. These studies demonstrated that msp4 sequences, but not MSP1a DNA or protein sequences, provide phylogeographic information and also that MSP1a sequences are highly heterogeneous among A. marginale populations. However, little information is available on the genetic diversity of A. marginale isolates from other regions of the world. The present study was undertaken to examine genetic variation among 10 isolates of A. marginale obtained from infected cattle in the State of Minas Gerais, Brazil, where A. marginale is endemic. Neighbor-joining analysis of msp4 sequences of Brazilian and New World isolates of A. marginale from Argentina, Mexico and the US provided bootstrap support for a Latin American clade. The sequences of the MSP1a repeats of four Brazilian isolates of A. marginale were compared to sequences of Latin American and US isolates. The MSP1a repeated sequences of Latin American isolates of A. marginale had nine repeat forms, alpha-phi, which have not been reported previously in North American isolates of A. marginale. Furthermore, the repeated forms tau, sigma and mu were only present in the Brazilian isolates. The results demonstrated that the genetic heterogeneity observed among isolates of A. marginale is common in endemic areas, independent of the predominant tick vector and is consistent with previous studies in which msp4 provided phylogeographic information about A. marginale isolates, while MSP1a was found not to be a useful marker for phylogeographic characterization of A. marginale isolates.


Asunto(s)
Anaplasma marginale/genética , Anaplasmosis/parasitología , Enfermedades de los Bovinos/parasitología , Secuencia de Aminoácidos , Anaplasma marginale/aislamiento & purificación , Animales , Secuencia de Bases , Brasil , Bovinos , ADN Protozoario/química , ADN Protozoario/genética , Variación Genética , Datos de Secuencia Molecular , Filogenia , Reacción en Cadena de la Polimerasa/veterinaria , Alineación de Secuencia
15.
Vaccine ; 32(19): 2160-6, 2014 Apr 17.
Artículo en Inglés | MEDLINE | ID: mdl-24606864

RESUMEN

Bovine anaplasmosis is a disease caused by the intraerythrocytic rickettsia Anaplasma marginale. Surface proteins (MSPs) of A. marginale are important in the interaction of the pathogen with the host and constitute potential vaccine targets against this pathogen. Currently, there is no commercial inactivated vaccine against bovine anaplasmosis that can generate a protective immune response that effectively prevents the development of clinical disease. The objective of this study was to evaluate the humoral and cellular immune responses of BALB/c mice immunized with the recombinant fragment of rMSP1a from A. marginale using carbon nanotubes as a carrier molecule. The fragment of rMSP1a comprising the N-terminal region of the protein was expressed in Escherichia coli BL21, purified by nickel affinity chromatography and covalently linked to multiwalled carbon nanotubes (MWNTs). After this functionalization, thirty BALB/c mice were divided into five groups, G1 (rMSP1a), G2 (MWNT+rMSP1a), G3 (MWNT), G4 (adjuvant) and G5 (unimmunized). The mice were immunized subcutaneously at days 0, 21 and 42. Blood samples were collected on day 11 after immunization. The spleens were collected, and the splenocytes were cultured for cell proliferation assays and cell immunophenotyping. Mice immunized with rMSP1a (G1 and G2) produced high levels of anti-rMSP1a IgG, demonstrating that the functionalization to carbon nanotubes did not interfere with protein immunogenicity. Immunization with MWNT+rMSP1a significantly induced higher percentages of CD4(+)CD44(+) and CD4(+)CD62L(+) lymphocytes, high levels of TNF-α, and a higher proliferative rate of splenocytes compared to mice immunized with rMSP1a alone (G1 group). Therefore, additional experiments using cattle should be performed to determine the efficacy, safety, immunogenicity and protection induced by rMSP1a associated with MWNT.


Asunto(s)
Anaplasmosis/prevención & control , Proteínas de la Membrana Bacteriana Externa/inmunología , Vacunas Bacterianas/inmunología , Nanotubos de Carbono/química , Anaplasma marginale , Animales , Anticuerpos Antibacterianos/sangre , Vacunas Bacterianas/química , Células Cultivadas , Citocinas/inmunología , Portadores de Fármacos/química , Epítopos/inmunología , Femenino , Inmunidad Celular , Inmunidad Humoral , Ratones Endogámicos BALB C , Proteínas Recombinantes/inmunología , Bazo/citología , Bazo/inmunología , Linfocitos T/inmunología
16.
Ticks Tick Borne Dis ; 4(4): 341-5, 2013 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-23567028

RESUMEN

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.


Asunto(s)
ADN Protozoario/genética , Ciervos , Infecciones Protozoarias en Animales/sangre , Animales , Brasil/epidemiología , Femenino , Masculino , Infecciones Protozoarias en Animales/epidemiología , Infecciones Protozoarias en Animales/parasitología
17.
Pesqui. vet. bras ; 38(5): 896-901, May 2018. tab
Artículo en Portugués | LILACS, VETINDEX | ID: biblio-955418

RESUMEN

O objetivo desse estudo foi investigar a ocorrência de tripanossomose em uma propriedade leiteira no município de Timon no estado do Maranhão, Brasil. O proprietário relatava histórico de abortos, nascimentos de crias fracas e mortalidade de animais adultos com perda progressiva de peso. Foram realizadas visitas à propriedade para obtenção do histórico, exame dos animais e coleta de sangue para realização do teste de Woo, hemogramas, testes sorológicos para pesquisa de anticorpos contra tripanossomose, leptospirose, e neosporose e PCR para diagnóstico molecular de Trypanosoma vivax. A identificação de animais com baixos valores no hematócrito foi a principal alteração hematológica identificada no rebanho. Dois animais foram positivos no teste de Woo, sendo visualizados tripanossomas em esfregaços sanguíneos, confirmados por meio de diagnóstico molecular como sendo T. vivax. Identificou-se que 95,23% (40/42) dos animais com hematócrito baixo foram sorologicamente positivos para T. vivax. As condições identificadas na propriedade, como ambiente propício aos vetores mecânicos, a presença de animais silvestres e a introdução de animais de estados onde já haviam sido registrados surtos de tripanossomose provavelmente estiveram associadas à introdução e disseminação do agente no rebanho. O elevado número de animais sorologicamente positivos para tripanossomose 82,51% (151/183) demonstra que praticamente todo o rebanho teve contato com o agente. O rápido estabelecimento das medidas de controle, entre elas a utilização das drogas tripanocidas, contribuiu para o controle do surto. O estudo permitiu comprovar a ocorrência de mais um surto de tripanossomíase tripanossomose no Brasil. O diagnóstico clínico da enfermidade foi dificultado pela semelhança dos sinais clínicos com outras enfermidades e pela possibilidade da associação de duas ou mais doenças no mesmo paciente, o que ressalta a importância do estabelecimento de medidas diagnósticas adequadas como forma de evitar a disseminação da enfermidade e minimizar as perdas econômicas dos produtores.(AU)


The objective of this study was to investigate the occurrence of trypanosomiasis in a dairy farm in municipality of Timon, State of Maranhão, Brazil. The owner reported abortus, births of weak calves, and mortality of adult animals with progressive weight loss. Visits to the property were carried out to obtain the history, realize animal examination and blood collection for the Woo test, hemograms, serological tests for trypanosomiasis, leptospirosis, and neosporosis and PCR for molecular diagnosis of Trypanosoma vivax. The identification of animals with low values in the hematocrit was the main hematological alteration identified in the herd. Two animals were positive in the Woo test and trypanosomes were visualized in blood smears, confirmed by molecular diagnosis as T. vivax. It was identified that 95.23% (40/42) of the animals with low hematocrit were serologically positive for T. vivax. The conditions identified in the property as an environment propitious to mechanical vectors, the presence of wild animals and the introduction of animals from states where trypanosomiasis outbreaks had already been reported were probably associated with the introduction and dissemination of the agent in the herd. The high number of serologically positive animals for trypanosomiasis 82.51% (151/183) shows that almost all the herd had contact with the agent. The rapid establishment of control measures, including the use of trypanocidal drugs, contributed to the control of the outbreak. The study allowed confirming the occurrence of another outbreak of trypanosomiasis in Brazil. The clinical diagnosis of the disease was difficult by the similarity of the clinical signs of trypanosomiasis with other diseases and the possibility of association of two or more diseases in the same patient, which emphasizes the importance of establishing adequate diagnostic measures as a way to avoid the dissemination of the disease and to minimize the economic losses of the producers.(AU)


Asunto(s)
Animales , Bovinos , Tripanosomiasis Bovina/diagnóstico , Tripanosomiasis Bovina/epidemiología , Bovinos/parasitología , Trypanosoma vivax/patogenicidad
18.
PLoS One ; 8(6): e65243, 2013.
Artículo en Inglés | MEDLINE | ID: mdl-23776456

RESUMEN

Bovine anaplasmosis is caused by cattle infection with the tick-borne bacterium, Anaplasma marginale. The major surface protein 1a (MSP1a) has been used as a genetic marker for identifying A. marginale strains based on N-terminal tandem repeats and a 5'-UTR microsatellite located in the msp1a gene. The MSP1a tandem repeats contain immune relevant elements and functional domains that bind to bovine erythrocytes and tick cells, thus providing information about the evolution of host-pathogen and vector-pathogen interactions. Here we propose one nomenclature for A. marginale strain classification based on MSP1a. All tandem repeats among A. marginale strains were classified and the amino acid variability/frequency in each position was determined. The sequence variation at immunodominant B cell epitopes was determined and the secondary (2D) structure of the tandem repeats was modeled. A total of 224 different strains of A. marginale were classified, showing 11 genotypes based on the 5'-UTR microsatellite and 193 different tandem repeats with high amino acid variability per position. Our results showed phylogenetic correlation between MSP1a sequence, secondary structure, B-cell epitope composition and tick transmissibility of A. marginale strains. The analysis of MSP1a sequences provides relevant information about the biology of A. marginale to design vaccines with a cross-protective capacity based on MSP1a B-cell epitopes.


Asunto(s)
Proteínas de la Membrana Bacteriana Externa/química , Proteínas de la Membrana Bacteriana Externa/inmunología , Regiones no Traducidas 5'/genética , Animales , Proteínas de la Membrana Bacteriana Externa/genética , Bovinos , Biología Computacional , Epítopos de Linfocito B/genética , Genotipo , Repeticiones de Microsatélite/genética , Estructura Secundaria de Proteína , Secuencias Repetidas en Tándem/genética
19.
Pesqui. vet. bras ; 37(1): 1-7, jan. 2017. tab.
Artículo en Portugués | LILACS, VETINDEX | ID: biblio-837442

RESUMEN

Este estudo objetivou determinar a soroprevalência da Babesiose e Anaplasmose em bovinos dos municípios de Ouricuri e Petrolina, estado de Pernambuco, Brasil; e definir os possíveis fatores de risco para a ocorrência dessas doenças. Amostras de sangue foram coletadas para realização de teste sorológico por Imunofluorescência Indireta (RIFI). Questionários epidemiológicos sanitários foram aplicados aos produtores com o objetivo de identificar possíveis fatores de risco. Carrapatos foram coletados, identificados e testados por Reação em Cadeia da Polimerase (PCR) para o diagnóstico da infecção por Anaplasma marginale, Babesia bigemina e Babaesia bovis. O estudo foi conduzido com 861 bovinos, sendo 468 de Petrolina e 393 de Ouricuri. A soroprevalência de A. marginale, B. bigemina e B. bovis em Petrolina foi de 35,0% (164/468), 35,9% (168/468) e 32,3% (151/468), respectivamente; e em Ouricuri foi de 45,5% (179/393), 38,6% (152/393) e 54,9% (216/393), respectivamente. A co-infecção por Anaplasma spp. e Babesia spp. foi observada em 31,6% e 32,1% de amostras de Petrolina e Ouricuri, respectivamente. A detecção de DNA de Babesia spp. por PCR foi possível em 5,8% (8/137) carrapatos, dos quais em 62,5 % (5/8) foi detectada posteriormente infecção por B. bovis, e em 23,3% (32/137) por A. marginale. A presença de carrapatos, o uso de acaricidas, idade, raça, e o município de residência dos animais foram identificados como fatores de risco para TPB pela análise univariável e multivariável. Este estudo permitiu caracterizar os municípios estudados como de instabilidade enzoótica para esses hemoparasitos, e consequentemente, alertar para adoção de medidas adequadas de controle e realização de novos estudos.(AU)


This study aimed to determine the seroprevalence of Babesiosis and Anaplasmosis in cattle from the municipalities of Ouricuri and Petrolina, state of Pernambuco, Brazil, and to define the risk factors for the occurrence of the diseases. Blood samples were collected for serologic testing by Indirect Immunofluorescence Assay (IFA). Sanitary epidemiological questionnaires were applied to the producers aiming to identify possible risk factors. Ticks were collected, identified and tested by Polymerase Chain Reaction (PCR) for the diagnosis of infection by Anaplasma marginale, Babesia bigemina and Babesia bovis. The study was conducted with 861 cattle, being 468 in Petrolina and 393 in Ouricuri. The seroprevalence of A. marginale, B. bigemina and B. bovis in Petrolina was of 35.0% (164/468), 35.9% (168/468) and 32.3% (151/468), respectively; and in Ouricuri was 45.5% (179/393), 38.6% (152/393), and 54.9% (216/393), respectively. Co-infection for Anaplasma spp. and Babesia spp. was observed in 31.6% and 32.1% samples of Petrolina and Ouricuri, respectively. The detection of DNA of Babesia spp. by PCR was possible in 5.8% (8/137) ticks; which 62.5% (5/8) was detected later infection with B. bovis; and 23.3% (32/137) with A. marginale. The presence of ticks, the use of acaricide, age, race, and county of residence of the animals were identified as risk factors for TBD by univariate analysis and multivariate. This study allowed the characterization of the municipalities studied as enzootic instability areas for these hemoparasitic, and consequently alert for adoption of adequate control measures and new studies.(AU)


Asunto(s)
Animales , Bovinos , Anaplasmosis/sangre , Anaplasmosis/epidemiología , Anticuerpos/análisis , Babesiosis/sangre , Babesiosis/epidemiología , Estudios Seroepidemiológicos , Encuestas Epidemiológicas
20.
Pesqui. vet. bras ; 37(1): 01-07, jan. 2017. tab
Artículo en Portugués | LILACS, VETINDEX | ID: biblio-1487602

RESUMEN

This study aimed to determine the seroprevalence of Babesiosis and Anaplasmosis in cattle from the municipalities of Ouricuri and Petrolina, state of Pernambuco, Brazil, and to define the risk factors for the occurrence of the diseases. Blood samples were collected for serologic testing by Indirect Immunofluorescence Assay (IFA). Sanitary epidemiological questionnaires were applied to the producers aiming to identify possible risk factors. Ticks were collected, identified and tested by Polymerase Chain Reaction (PCR) for the diagnosis of infection by Anaplasma marginale, Babesia bigemina and Babesia bovis. The study was conducted with 861 cattle, being 468 in Petrolina and 393 in Ouricuri. The seroprevalence of A. marginale, B. bigemina and B. bovis in Petrolina was of 35.0% (164/468), 35.9% (168/468) and 32.3% (151/468), respectively; and in Ouricuri was 45.5% (179/393), 38.6% (152/393), and 54.9% (216/393), respectively. Co-infection for Anaplasma spp. and Babesia spp. was observed in 31.6% and 32.1% samples of Petrolina and Ouricuri, respectively. The detection of DNA of Babesia spp. by PCR was possible in 5.8% (8/137) ticks; which 62.5% (5/8) was detected later infection with B. bovis; and 23.3% (32/137) with A. marginale. The presence of ticks, the use of acaricide, age, race, and county of residence of the animals were identified as risk factors for TBD by univariate analysis and multivariate. This study allowed the characterization of the municipalities studied as enzootic instability areas for these hemoparasitic, and consequently alert for adoption of adequate control measures and new studies.


Este estudo objetivou determinar a soroprevalência da Babesiose e Anaplasmose em bovinos dos municípios de Ouricuri e Petrolina, estado de Pernambuco, Brasil; e definir os possíveis fatores de risco para a ocorrência dessas doenças. Amostras de sangue foram coletadas para realização de teste sorológico por Imunofluorescência Indireta (RIFI). Questionários epidemiológicos sanitários foram aplicados aos produtores com o objetivo de identificar possíveis fatores de risco. Carrapatos foram coletados, identificados e testados por Reação em Cadeia da Polimerase (PCR) para o diagnóstico da infecção por Anaplasma marginale, Babesia bigemina e Babaesia bovis. O estudo foi conduzido com 861 bovinos, sendo 468 de Petrolina e 393 de Ouricuri. A soroprevalência de A. marginale, B. bigemina e B. bovis em Petrolina foi de 35,0% (164/468), 35,9% (168/468) e 32,3% (151/468), respectivamente; e em Ouricuri foi de 45,5% (179/393), 38,6% (152/393) e 54,9% (216/393), respectivamente. A co-infecção por Anaplasma spp. e Babesia spp. foi observada em 31,6% e 32,1% de amostras de Petrolina e Ouricuri, respectivamente. A detecção de DNA de Babesia spp. por PCR foi possível em 5,8% (8/137) carrapatos, dos quais em 62,5 % (5/8) foi detectada posteriormente infecção por B. bovis, e em 23,3% (32/137) por A. marginale. A presença de carrapatos, o uso de acaricidas, idade, raça, e o município de residência dos animais foram identificados como fatores de risco para TPB pela análise univariável e multivariável. Este estudo permitiu caracterizar os municípios estudados como de instabilidade enzoótica para esses hemoparasitos, e consequentemente, alertar para adoção de medidas adequadas de controle e realização de novos estudos.


Asunto(s)
Animales , Bovinos , Anaplasmosis/epidemiología , Anaplasmosis/sangre , Anticuerpos/análisis , Babesiosis/epidemiología , Babesiosis/sangre , Estudios Seroepidemiológicos , Encuestas Epidemiológicas
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