Diss. (sammanfattning) Uppsala : Sveriges lantbruksuniv., 2008. / Härtill 4 uppsatser.

Thesis (Ph. D.)--Washington State University, December 2008. / Title from PDF title page (viewed on Apr. 29, 2010). "College of Veterinary Medicine." Includes bibliographical references.

Thesis (M.S.)--Humboldt State University, 2006. / Includes bibliographical references (leaves 40-46). Also available via Humboldt Digital Scholar.

Background: Equine granulocytic anaplasmosis (EGA), caused by the organism Anaplasma phagocytophilum, is a tick-borne disease of clinical importance in Southwest Virginia. The disease is recognized worldwide and causes pyrexia, anorexia, limb edema, and lethargy. Diagnosis in endemic areas is often based on clinical signs, but confirmation of infection can be made via detection of morulae on a peripheral blood smear or polymerase chain reaction analysis (PCR) at the time of disease or by serologic detection of antibodies 2-4 weeks post infection. There is growing interest in stall-side methods for diagnosis of various equine diseases which has led to an increased use of the SNAP 4DX Plus Test® for vector-borne diseases. Objectives: Determine seroprevalence of antibodies to A. phagocytophilum in the equine population of Southwest Virginia and changes in seroprevalence compared to samples taken 6 years earlier. Determine the percentage of horses with clinical signs consistent with EGA that were positive for A. phagocytophilum infection and assess common presenting clinical signs, hematologic variables, and confirmatory diagnostic test results. Animals: Seroprevalence was evaluated in horses presented for routine annual Coggins testing in 2013 and 2019-2020. Clinical features of disease and diagnostic test results were evaluated in horses presenting with clinical signs compatible with A. phagocytophilum infection from September 2019-August 2020. Methods: Seroprevalence was determined using the IDEXX SNAP 4DX Plus Test® on serum collected from horses presenting for annual Coggins testing in 2013 and 2019-2020. Samples collected in 2013 had been stored at -7580 degrees F since collection. Age, sex, county of residence, and month of sampling were statistically analyzed in the seroprevalence population. Horses presenting with clinical disease consistent with EGA from September 2019-August 2020 had the following diagnostic tests performed: complete blood count (CBC), blood smear for morulae detection, polymerase chain reaction (PCR) analysis, immunofluorescence antibody testing (IFAT), and the IDEXX SNAP 4DX Plus Test®. Results: Seroprevalence of A. phagocytophilum in the equine population of Southwest Virginia increased from 8.5% in 2013 to 11.2% in 2019-2020, although this increase was not statistically significant. In the 2019-2020 population, month of sampling was significantly associated with presence of antibodies to A. phagocytophilum. Positive samples were more common from November-February than other times of the year. When the two sample time periods were combined, sex was significantly associated with presence of antibodies to A. phagocytophilum with geldings more likely to be seropositive. Within the clinical case population, 35% of horses with clinical signs compatible with equine granulocytic anaplasmosis had confirmed infection. The most common hematologic abnormality in affected horses was thrombocytopenia. PCR analysis was the most sensitive diagnostic test to diagnose infection followed by identification of morulae on blood smears. Conclusions: Seroprevalence of A. phagocytophilum is similar to other endemic areas in the United States and appears to be increasing over time. In active clinical cases, diagnosis is best made via PCR or detection of morulae on a blood smear. The SNAP 4DX Plus Test® was not appropriate for diagnosis of active EGA in acute cases. Seroprevalence of Anaplasma phagocytophilum in the equine population of Southwest Virginia / Master of Science / Equine granulocytic anaplasmosis (EGA) is a common tick-borne disease in the United States and worldwide. The causative bacteria, Anaplasma phagocytophilum, also infects humans, dogs, and various domestic animal species. In horses, signs of disease include fever, decreased appetite, leg swelling, and depression. Diagnostic testing that is both accurate and timely is still lacking. The point-of-care SNAP 4DX Plus Test® used to diagnose vector-borne infectious disease in dogs has been suggested for similar use in horses. The objectives of the current study were to determine seroprevalence of antibodies to A. phagocytophilum in the equine population of Southwest Virginia and to characterize the clinical signs and diagnostic test findings of horse with clinical signs of EGA. Seroprevalence was determined using the SNAP 4DX Plus Test®. Serum samples were obtained from horses presenting for annual Coggins testing in 2019-2020. Samples from 2013 were also tested to determine if seroprevalence had increased. Horses presenting with clinical signs consistent with A. phagocytophilum were examined by a veterinarian and had blood drawn for a complete blood count (CBC), blood smear evaluation, polymerase chain reaction analysis (PCR), immunofluorescent antibody testing (IFAT), and the SNAP 4DX Plus Test®. Seroprevalence in 2019-2020 was 11.2% and 8.5% in 2013. This is similar to other endemic areas in the United States and Europe. In horses sampled from 2019-2020, the month of sampling was significantly associated with presence of antibodies to A. phagocytophilum with most of the positive samples being identified in November through February. Geldings were more likely to be seropositive than mares. Thirty five percent of horses with signs consistent with EGA were confirmed to have the disease. Within this population, PCR analysis and/or detection of morulae on the blood smear were reliable indicators of disease while diagnostic techniques utilizing serology were unreliable. This is the first study to determine seroprevalence of A. phagocytophilum in Southwest Virginia. In the actively infected population, PCR and blood smear evaluation remain the most sensitive methods of diagnosis. While the SNAP 4DX Plus Test® is useful for serologic data collection, it was not appropriate for acute diagnosis of EGA.

Anaplasma phagocytophilum and A. marginale are the etiologic agents of human granulocytic anaplasmosis and bovine anaplasmosis, respectively. Both diseases can be severe, even fatal, and protective vaccines for each are lacking. We recently identified A. phagocytophilum outer membrane protein A (ApOmpA) as being critical for cellular invasion and is expressed during infection of mammalian but not tick cells. Disrupting ApOmpA-host cell interactions significantly inhibits A. phagocytophilum entry into host cells. ApOmpA and its A. marginale ortholog, AM854 (A. marginale OmpA; AmOmpA) exhibit 44% amino acid identity. The ApOmpA invasin domain is highly conserved between both proteins. In this study, we investigated the differential expression of AmOmpA in mammalian versus tick cell lines; the serological cross-reactivity between AmOmpA and ApOmpA; the potential role of AmOmpA in mediating interactions with mammalian host cells; and if inhibiting the AmOmpA-host cell interaction impairs A. marginale cellular invasion. AmOmpA is expressed throughout infection of mammalian, but not tick cells. Sera from A. marginale infected cows recognized both AmOmpA and ApOmpA. Sera from cows immunized with an A. marginale OM complex that conferred protection also recognized both proteins. Thus, ApOmpA and AmOmpA share cross-reactive B-cell epitopes. To determine if AmOmpA plays a role in promoting A. marginale infection, we assessed the abilities of recombinant AmOmpA to competitively inhibit infection of mammalian host cells. To examine the cross-reactive properties of OmpA, we showed that preincubation of host cells with GST-ApOmpA and pretreatment of A. marginale with anti-GST-ApOmpA significantly inhibit A. marginale infection of host cells; and that pretreatment of A. phagocytophilum with serum from cows immunized with an A. marginale OM complex reduces its infection of host cells. These studies advance understanding of conservation of OmpA-mediated cellular invasion between Anaplasma species and highlight the potential of OmpA as a vaccinogen that could offer protection against human and veterinary anaplasmoses.

Em função da proximidade cada vez maior entre o cavalo e o homem, é de extrema importância ter conhecimentos das doenças que acometem os equinos, que por ventura, podem acometer seres humanos. Dentre muitas doenças, pode-se citar duas, que promovem grandes perdas econômicas aos rebanhos eqüinos, tanto no tratamento desses rebanhos, como com a morte dos mesmos, dificultando a importação e exportação de animais: a babesiose e a erliquiose (anaplasmose), que podem estar ou não associadas, acometendo um animal, concomitantemente. A presente pesquisa teve como objetivo investigar e diagnosticar doença concomitante babesiose (por Babesia equi ou Theileria equi) e Erliquiose (por Erliquia equi ou Anaplasma phagocytophilum), no estado de São Paulo, em rebanhos eqüinos, utilizando as técnicas de Nested PCR (Nested polymerase chain reaction reação em cadeia pela polimerase para diagnóstico de T. equi e A. phagocytophilum) e c-ELISA (competitive enzyme-linked immunosorbent assay para diagnóstico de T. equi) ou ELISA indireto (para diagnóstico de A. phagocytophilum) e comparar os resultados obtidos nas diferentes técnicas em 250 amostras de eqüino (sangue total e soro). Como resultado, obteve-se 38,4%, 46% e 36% de positividade, respectivamente, nos testes de pesquisa de hematozoário, c-ELISA e Nested PCR para Theileria equi e 0%, 3% e 0% de positividade, respectivamente, nos testes de pesquisa de hemoparasita, ELISA indireto e Nested PCR para Anaplasma phagocytophilum, não sendo observada a co-infecção de Babesiose e Anaplasmose no rebanho estudo / Due to the increasing proximity between horse and man, it is of extreme importance to understand the diseases that affect horses which by chance may affect humans. Among many diseases, babesiosis and ehrlichiosis (anaplasmosis) promote high economic losses to horses herds in consequence of costs of treatment and also death, making it difficult to import and export animals: They can or not be linked affecting an animal at the same time. This study aimed to investigate and diagnose concomitant babesiosis (Babesia equi and Theileria equi) and ehrlichiosis (for ehrlichia equipment or Anaplasma phagocytophilum) in equine herds of the state of Sao Paulo, using the techniques of Nested PCR (Nested polymerase chain reaction for the diagnosis of T. equi and A. phagocytophilum) and c-ELISA (competitive enzyme-linked immunosorbent assay for diagnosis of T. equi) or ELISA (for diagnosis of A. phagocytophilum). Also to compare results obtained in these different techniques in 250 samples of horse (whole blood and serum). Results showed 38.4%, 46% and 36% positivity, respectively, in tests for the detection of Theileria equi through hematozoan, c-ELISA and Nested PCR and 0%, 3% and 0% positivity, respectively, in tests for the detection of Anaplasma phagocytophilum through blood parasites, indirect ELISA and Nested PCR. It was not observed co-infection Babesiosis and anaplasmosis in the herd study

Em função da proximidade cada vez maior entre o cavalo e o homem, é de extrema importância ter conhecimentos das doenças que acometem os equinos, que por ventura, podem acometer seres humanos. Dentre muitas doenças, pode-se citar duas, que promovem grandes perdas econômicas aos rebanhos eqüinos, tanto no tratamento desses rebanhos, como com a morte dos mesmos, dificultando a importação e exportação de animais: a babesiose e a erliquiose (anaplasmose), que podem estar ou não associadas, acometendo um animal, concomitantemente. A presente pesquisa teve como objetivo investigar e diagnosticar doença concomitante babesiose (por Babesia equi ou Theileria equi) e Erliquiose (por Erliquia equi ou Anaplasma phagocytophilum), no estado de São Paulo, em rebanhos eqüinos, utilizando as técnicas de Nested PCR (Nested polymerase chain reaction reação em cadeia pela polimerase para diagnóstico de T. equi e A. phagocytophilum) e c-ELISA (competitive enzyme-linked immunosorbent assay para diagnóstico de T. equi) ou ELISA indireto (para diagnóstico de A. phagocytophilum) e comparar os resultados obtidos nas diferentes técnicas em 250 amostras de eqüino (sangue total e soro). Como resultado, obteve-se 38,4%, 46% e 36% de positividade, respectivamente, nos testes de pesquisa de hematozoário, c-ELISA e Nested PCR para Theileria equi e 0%, 3% e 0% de positividade, respectivamente, nos testes de pesquisa de hemoparasita, ELISA indireto e Nested PCR para Anaplasma phagocytophilum, não sendo observada a co-infecção de Babesiose e Anaplasmose no rebanho estudo / Due to the increasing proximity between horse and man, it is of extreme importance to understand the diseases that affect horses which by chance may affect humans. Among many diseases, babesiosis and ehrlichiosis (anaplasmosis) promote high economic losses to horses herds in consequence of costs of treatment and also death, making it difficult to import and export animals: They can or not be linked affecting an animal at the same time. This study aimed to investigate and diagnose concomitant babesiosis (Babesia equi and Theileria equi) and ehrlichiosis (for ehrlichia equipment or Anaplasma phagocytophilum) in equine herds of the state of Sao Paulo, using the techniques of Nested PCR (Nested polymerase chain reaction for the diagnosis of T. equi and A. phagocytophilum) and c-ELISA (competitive enzyme-linked immunosorbent assay for diagnosis of T. equi) or ELISA (for diagnosis of A. phagocytophilum). Also to compare results obtained in these different techniques in 250 samples of horse (whole blood and serum). Results showed 38.4%, 46% and 36% positivity, respectively, in tests for the detection of Theileria equi through hematozoan, c-ELISA and Nested PCR and 0%, 3% and 0% positivity, respectively, in tests for the detection of Anaplasma phagocytophilum through blood parasites, indirect ELISA and Nested PCR. It was not observed co-infection Babesiosis and anaplasmosis in the herd study

Submitted by Jorge Silva (jorgelmsilva@ufrrj.br) on 2018-10-19T18:19:28Z No. of bitstreams: 1 2013 - Andresa Guimar?es.pdf: 1221110 bytes, checksum: 80ca8363650edbb042caea761cb85083 (MD5) / Made available in DSpace on 2018-10-19T18:19:28Z (GMT). No. of bitstreams: 1 2013 - Andresa Guimar?es.pdf: 1221110 bytes, checksum: 80ca8363650edbb042caea761cb85083 (MD5) Previous issue date: 2013-08-21 / CAPES / CNPq / Studies of domestic cats are little discussed in Brazil, especially regarding hemoparasita research where cats can be carriers of the agent and contribute to infection of man. Anaplasmataceae agents are gram-negative bacteria found in leukocytes and platelets, in which morulae characteristic of the infection is observed. The transmission of these agents to humans and animals occurs by arthropod vectors. Sorologic diagnostic methods are emphasized, but recent studies using molecular techniques demonstrate greater sensitivity and specificity, with better characterization of the agent. The present study aims to diagnose domestic cats in Rio de Janeiro naturally infected by agents Anaplasmataceae (Ehrlichia sp., and Anaplasma sp.) by serological and molecular methods, as well as evaluate hematological changes associated with these infections to guide veterinarians in the diagnosis of these diseases. Samples of 216 animals from clinics located in cities in the metropolitan region of Rio de Janeiro were used. The research of antibodies IgG anti-E. canis was performed by Immunofluorescence Assay (IFAT) and molecular detection of Ehrlichia sp. by nested Polymerase Chain Reaction (PCR) based on 16S rRNA gene. The results demostrate that 18 (8.3%) animals were positive in the direct detection by blood smear, 57 (26.4%) seropositive and 37 (17.1%) positive by nested-PCR for Ehrlichia sp., being 91 (42.1%) positive in at least one technique. The diagnosis of A. platys was performed by real-time PCR (qPCR) and nested-PCR based on 16S rRNA and gltA genes, respectively. Seventeen animals (7.9%) showed inclusions in platelet, eight (3.7%) were positive in qPCR, two (0,9%) in nested-PCR and 23 (10.6%) cats were considered positive for A. platys in at least one technique. The detection of A. phagocytophilum was performed by qPCR based on gene msp2, however, positive animals were not observed. The most common hematologic findings in animals positive for agents Anaplasmataceae were thrombocytopenia, leukocytosis, neutrophilia, regenerative left shift, anemia and hyperproteinemia. Only one animal showed co-positivity for both agents. Regarding the associated factor, only age interfered with positivity for Ehrlichia sp., in wich younger IFAT positive animals were less prone to infection, unlike that observed in the nested-PCR in which the younger had higher positivity. Only two positive samples in the nested-PCR for E. canis were subjected to sequencing. One sequence demonstrated 100% similarity with isolates of Ehrlichia sp. and E. canis and another showed 99% similarity with isolates of Ehrlichia sp. ocelot and Ehrlichia sp. little-spotted-cat. The two positive samples in the nested-PCR for A. platys demonstrated 100% similarity with isolates of A. platys reported in advanced. The present work demonstrates the circulation of agents Anaplasmataceae in domestic cats in Rio de Janeiro. / Estudos com felinos dom?sticos s?o pouco abordados no Brasil, principalmente em rela??o ? pesquisa de hemoparasitoses, em que os gatos podem ser portadores do agente e contribuir para infec??o do homem. Os agentes Anaplasmataceae s?o bact?rias gram-negativas que s?o encontradas em leuc?citos, plaquetas ou eritr?citos, formando m?rulas, caracter?sticas de sua infec??o. A transmiss?o desses agentes para o homem e animais ocorre principalmente por vetores artr?podes. Como m?todo diagn?stico ressalta-se a sorologia, por?m estudos recentes utilizando diagn?stico molecular demonstram maior sensibilidade e especificidade, com melhor caracteriza??o do agente. O presente estudo tem como objetivo diagnosticar felinos dom?sticos do Rio de Janeiro infectados naturalmente por agentes Anaplasmataceae (Ehrlichia sp. e Anaplasma sp.) por meio de m?todos sorol?gico e molecular, bem como avaliar as altera??es hematol?gicas associadas a estas infec??es. Foram utilizadas amostras de 216 animais, provenientes de cl?nicas localizadas em cidades da Regi?o Metropolitana do Rio de Janeiro. A pesquisa de anticorpos IgG anti-E. canis foi realizada pela Rea??o de Imunofluoresc?ncia Indireta (RIFI) e a detec??o molecular de Ehrlichia sp., pela nested Rea??o em Cadeia da Polimerase (nested-PCR) baseada no gene 16s rRNA. Os resultados demonstraram que 18 (8,3%) animais foram positivos na detec??o direta pelo esfrega?o sangu?neo, 57 (26,4%) soropositivos e 37 (17,1%) positivos na nested-PCR para Ehrlichia sp., sendo 91 (42,1%) considerados positivos para Ehrlichia sp. em pelo menos uma t?cnica. O diagn?stico de A. platys foi efetuado pela PCR em tempo real (qPCR) e nested-PCR baseada nos genes gltA e 16S rRNA, respectivamente. Dezessete animais (7,9%) apresentaram inclus?es em plaquetas, oito (3,7%) foram positivos na qPCR, dois (0,9%) na nested-PCR e 23 (10,6%) animais foram considerados positivos para A. platys em pelo menos uma t?cnica. A pesquisa de A. phagocytophilum foi realizada por meio da qPCR baseada no gene msp2; no entanto, n?o foram observados animais positivos. Os achados hematol?gicos mais frequentes nos animais positivos para agentes Anaplasmataceae foram trombocitopenia, leucocitose, neutrofilia, desvio ? esquerda regenerativo, anemia e hiperproteinemia. Apenas um animal apresentou co-positividade para ambos os agentes. Em rela??o aos dados associados, somente a idade interferiu na positividade para Ehrlichia sp., sendo na RIFI os animais mais jovens menos predispostos ? infec??o, ao contr?rio do observado na nested-PCR em que os mais jovens apresentaram maior ?ndice de positividade. Apenas duas amostras positivas na nested-PCR para E. canis foram submetidas ao sequenciamento, sendo que uma sequ?ncia apresentou 100% de similaridade com isolados de Ehrlichia sp. e E. canis e a outra demonstrou 99% de similaridade com isolados de Ehrlichia sp. ocelot e Ehrlichia sp. little-spotted-cat. As duas amostras positivas na nested-PCR para A. platys apresentaram 100% de similaridade com isolados de A. platys previamente descritos. O presente trabalho demonstra, portanto, a ocorr?ncia de agentes Anaplasmataceae na popula??o de felinos dom?sticos do Rio de Janeiro.

Thesis (Ph.D.)--Murdoch University, 2006. / Thesis submitted to the Division of Health Sciences. Bibliography: leaves 225-252.

Thesis--Auburn University, 2005. / Abstract. Vita. Includes bibliographic references.