Diagnosis Of Anaplasmosis Research Articles

Immunosensor for bovine anaplasmosis diagnosis on graphite platform functionalized with poly (3-hydroxybenzoic acid)

This study describe the development of an electrochemical immunosensor with a transducer platform functionalized with the poly-(3- hydroxybenzoic acid), to detection of antibodies against the surface protein (Am1) of Anaplasma marginale. This biological probe was immobilized on the polymer under graphite and characterized electrochemically. The detections of antigen-antibody interactions were conducted using the signal obtained from the oxidation of 4-aminoantipyrine (4 -AAP), interleaved with Am1, to enable the use of voltammetry technique of differential pulse. The detections of antibodies of A. marginale, using the graphite electrode functionalized with the probe Am1 was 66% higher than the graphite electrode not functionalized. The time of storage of the immunosensor was satisfactory, with reduced peak current by 35%, after 90 days. The results demonstrate the excellent applicability of this new functionalized platform for probe immobilization and diagnosis of anaplasmosis without the interference of the most similar bovine diseases

Evaluation of an Indirect Immunofluorescence Assay for the Detection of Anaplasma phagocytophilum Antigen in Ovine Buffy Coat Smears.

Diagnosis of anaplasmosis is challenging considering the great variation in clinical signs and the limitations of the available diagnostic assays, while the detection of carrier animals that play a significant role in disease epidemiology as reservoirs is of great significance. In this study, we evaluated the diagnostic accuracy of a newly developed indirect immunofluorescent assay (Ag-IFAT) for the detection of A. phagocytophilum antigens in buffy coat specimens, alone and in combination with cytology, using PCR as a reference. Blood samples were collected from 138 sheep of the Chios breed from six farms in Greece. A buffy coat was extruded from the centrifuged blood. Buffy coat smears were used for cytological examination and the Ag-IFAT assay. The Ag-IFAT assay presented excellent specificity (100%) and high sensitivity (85.4%) for the detection of A. phagocytophilum antigens in buffy coats, and it has an almost perfect agreement with PCR and cytology (κ value = 0.88 and 0.85, respectively). A. phagocytophilum antigens are likely to be detected using Ag-IFAT in a PCR-positive animal, as indicated by the good performance of the assay. Overall, this assay presents high diagnostic accuracy, and it could be used for the detection of animals during the early stage of infection.

Pathomorphological changes in the testicles of goats in acute anaplasmosis

The aim of the research was to study pathomorphological changes in the testicles of goats in acute anaplasmosis. Diagnosis of anaplasmosis in sick goats was carried out taking into account the characteristic clinical signs and studies of peripheral blood smears. Blood smears after fixation in alcohol ether were stained according to Romanovsky-Giemsa by hematoxilin and eosin. For histological examination, pieces of testicles and appendages of testicles of 0.5 cm thick were selected, which were fixed in an 8% neutral aqueous solution of formalin. The resulting material was poured into paraffin according to the generally accepted method. Histological sections of 5-8 microns thick were made from the obtained paraffin blocks and stained with hematoxilin and eosin using the method of Van Gieson and Mallory. Micro photos were made with a series of rendered images on the basis of Olimpus 2000. The study revealed the following changes: the spermatogenic epithelium has desquamation, in the lumen of the convoluted seminiferous, there were tubules protein detritus, exfoliated cells of seminiferous epithelium and macrophages. Sertolli cells were in a state of vacuole dystrophy and necrosis. In the interstitial tissue around arterial vessels and convoluted seminiferous tubules, focal accumulation of lymphoid-macrophage infiltration was found. Endocrine cells located in the interstitial were in a state of vacuole dystrophy and necrosis. Thus, in goats older than 6 months with acute anaplasmosis in the testicles, pathomorphological changes are characteristic of acute parenchymal orchitis.

Determination of Diagnostic Value of cELISA for the Diagnosis of Anaplasmosis in Clinically Suspected Ruminants

The aim of this study was to determine diagnostic value of cELISA in anaplasmosis in clinically suspected animals and to compare the cELISA results with the clinical examination results. For this purpose a total of 720 ruminants (457 cattle, 146 sheep, 117 goat) were examined in terms of clinical signs. Eighty-eight ruminants consisting of 61 cattle, 11 sheep and 16 goat which had the symptoms of anemia, fever, icterus, weakness, depression and lack of appetite were selected for the study. Blood samples were collected from the jugular vein of all clinically suspected animals and serum samples were separated. A commercially available competitive enzyme-linked immunosorbent assay (C-ELISA) kit was used for determine antibodies to Anaplasma species. cELISA based diagnosis revealed that 47 of 88 serum samples (53.4%) were positive for anaplasmosis. In serological examination Anaplasma specific antibodies were determined in 45.9% of cattle, 63.6% of sheep and 56.2% of goats. Seropositivity rate was statistically differ among the age groups of cattle and the highest seropositvity rate was found in <12 month age (P<0.005). However no difference was found in the seropositivity rate of Anaplasma in sheep and goat in relation to age group. From the data obtained in this study it can be concluded that clinical findings are not sufficient criteria for the diagnosis of anaplasmosis and must be supported by serological examination.

English

Infection with Anaplasma phagocytophilum is of importance in dogs. It is called canine anaplasmosmosis and causes persistent clinical signs for long periods of time. Its main vector is a tick of the genus Ixodes. Diagnosis of anaplasmosis can be achieved either by serological or molecular methods. Presence of the disease has been reported in the U.S.A., Europe and Asia, but little is known about its prevalence in Mexico. In the present study, the enzyme-linked immunosorbent assay (ELISA) diagnostic method was used to determine the prevalence of antibodies against A. phagocytophilum at the city of Monterrey, Mexico. A total of 391 dogs were tested using a commercial kit. A 3% prevalence was found that included dogs of the breeds Doberman, French Puddle and Maltese, as well as mixed breed. &nbsp; Key words: Anaplasma phagocytophilum, dogs, Mexico.

PP-241 Pay attention to differential diagnosis of anaplasmosis with thrombocytopenic syndrome

PP-241 Pay attention to differential diagnosis of anaplasmosis with thrombocytopenic syndrome

OL-049 Evaluation on external quality control for diagnosis of Anaplasmosis in China

OL-049 Evaluation on external quality control for diagnosis of Anaplasmosis in China

Comparação entre diversos antígenos para o diagnóstico de Anaplasma marginale por ELISA

Anaplasmose bovina é uma doença com grande importância nas regiões tropicais e subtropicais do mundo por determinar perdas econômicas devido à mortalidade e redução da produtividade. É causada por Anaplasma marginale, uma riquétsia intraeritrocítica obrigatória cujo controle requer, além de uma vacina eficiente, uma acurada identificação de bovinos cronicamente infectados. Apesar de existirem atualmente diversos métodos de diagnóstico dessa riquétsia, os métodos sorológicos, em particular o ensaio de imunoadsorção enzimática-ELISAs, são os mais utilizados devido à sua versatilidade e praticidade. No entanto, devido ao grande número de antígenos disponíveis, atualmente torna-se necessária uma avaliação para definir quais antígenos apresentam um melhor desempenho no diagnóstico da anaplasmose. Soros de bovinos positivos e negativos para A. marginale por PCR, e soros de animais provenientes do Brasil e Costa Rica, foram testados em ELISAs baseados em MSP1a, MSP2 e MSP5 recombinantes, um pool das três proteínas recombinantes, e antígeno de lisado de corpúsculos iniciais da riquétsia (CI). Utilizando soro de bovinos positivos para A. marginale por PCR, uma maior sensibilidade foi observada no ELISA CI. No entanto, uma maior especificidade, com soro de bovinos negativos a PCR, foi observada com os ELISAs recombinantes. O porcentual de bovinos positivos do Brasil e Costa Rica foi maior com ELISA CI. Razões para essas diferenças são discutidas.

Diagnosis of Anaplasmosis in Formalin-Fixed Tissue Using the Wolbach's Giemsa Stain

Bovine anaplasmosis is an economically significant disease that is widespread throughout the world and enzootic in 40 of the 50 states of the USA. 2 The causative agent, Anaplasma marginale, can be transmitted from infected blood by arthropod and mechanical vectors. Cattle of all ages can be infected, but severe clinical signs and death are common only in mature cattle. Clinical disease is due to anemia resulting from the intracellular destruction of parasitized erythrocytes by phagocytes. In fatal cases severe anemia, icterus, and splenomegaly are typical. Histologically, centrilobular hypoxic hepatic necrosis and splenic hemosiderosis related to erythrocyte destruction are commonly present. Diagnosis is established by identification of anaplasma bodies in the margins of erythrocytes using Giemsa or Wright-Giemsa stains or fluorescent antibody examination of blood smears or tissue impressions.’ Field necropsy specimens submitted to the California Veterinary Diagnostic Laboratory occasionally include samples from cattle that have a clinical history and histologic lesions of hypoxic liver damage or splenic hemosiderosis suggestive

The use of spleen antigen in the tube agglutination test for diagnosis of anaplasmosis in cattle.

An antigen was prepared from the spleen of an infected bovine calf and was used in the tube agglutination test. Cases having rare and very low parasitaemia as well as the carrier cases were detected by this procedure and no non-specific reaction was observed with other haemoprotozoan infections. The maximum titre obtained with the Anplasma positive sera was 1:20, about 60 per cent showed a titre of 1:10 and 90 per cent showed a titre of 1:5 or more. The test provided satisfactory results in detecting the cases even at the early phase of the infection. The test also proved useful in judging the efficacy of a drug employed to destroy the carrier status.