scholarly journals Serodiagnosis of Babesia equi in horses submitted to exercise stress

2007 ◽  
Vol 27 (4) ◽  
pp. 179-183 ◽  
Author(s):  
Cristiane D. Baldani ◽  
Rosangela Z. Machado ◽  
Tânia F. Raso ◽  
Aramis A. Pinto
Keyword(s):  
High Speed ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Exercise Stress ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Babesia Equi ◽  
Combined Use ◽  
Before And After ◽  
Disease Free

A complement fixation test (CFT), performed in microtitre plates, based upon the use of crude antigenic preparation of Babesia equi was adapted for the detection of antibodies in serum of infected horses. The indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were also used for the immunodiagnosis of B. equi. Serum samples from 15 apparently healthy horses, previously conditioned to a high-speed equine treadmill, were taken before and after exercise. All the samples analyzed were positive for B. equi infection. There were no significant differences (P<0.01) between these 3 tests, or the condition of rest or stress. The combined use of CFT and IFAT or ELISA should be recommended in order to enable veterinary services to more efficiently prevent introduction of infected horses into disease-free areas.

scholarly journals Novel Indirect Fluorescent Antibody Test for Lyme Disease

1996 ◽  
Vol 8 (2) ◽  
pp. 196-201 ◽  
Author(s):  
Margaret A. Chambers ◽  
Larry J. Swango ◽  
James C. Wright
Keyword(s):  
Human Error ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Cross Reactivity ◽  
Enzyme Linked Immunosorbent Assay ◽  
Indirect Fluorescent Antibody ◽  
Serum Samples ◽  
Specific Pathogen ◽  
Membrane Bound ◽  
Animal Use

An indirect fluorescent antibody (IFA) test was developed using a novel format of Borrelia burgdorferi organisms adhered to a monolayer of cultured endothelial cells derived from an equine tumor. Sensitivity and specificity of the new IFA test for detecting anti- B. burgdorferi antibodies were evaluated using sera from dogs inoculated with live B. burgdorferi or vaccinated with B. burgdorferi bacterin or leptobacterins and from unvaccinated specific-pathogen-free (SPF) dogs. To compare the new IFA test with existing tests, serum samples were submitted to independent laboratories to be tested by enzyme-linked immunosorbent assay (ELISA) and a traditional IFA test. Samples were also tested with 2 commercially available membrane-bound ELISA kits. Both Borrelia-inoculated dogs and dogs vaccinated with B. burgdorferi bacterin developed levels of antibody detectable by the new IFA test. Dogs vaccinated with a combination canine vaccine or leptobacterin for food animal use developed detectable levels of antibody against Leptospira but remained seronegative for Borrelia by the new IFA test, as did the unvaccinated SPF dogs. The new IFA test was sensitive, detecting antibodies against B. burgdorferi as early as 7 days postinoculation. It was also specific, showing no cross-reactivity with anti- Leptospira antibodies induced by vaccination with leptobacterins. The new IFA test compared favorably with both the standardized traditional IFA test and ELISA. Results from both membrane-bound ELISA kits were not consistent when compared with each other or with the new IFA test. The new IFA test had low nonspecific fluorescence, which made it easier to evaluate and reduced the human error and variability of test results.

scholarly journals Detection of Immunoglobulin G Antibodies to Neospora caninum in Humans: High Seropositivity Rates in Patients Who Are Infected by Human Immunodeficiency Virus or Have Neurological Disorders

2006 ◽  
Vol 13 (1) ◽  
pp. 84-89 ◽  
Author(s):  
Janaína Lobato ◽  
Deise A. O. Silva ◽  
Tiago W. P. Mineo ◽  
Jodi D. H. F. Amaral ◽  
Gesmar R. Silva Segundo ◽  
...  
Keyword(s):  
Human Immunodeficiency Virus ◽  
Immunoglobulin G ◽  
Neurological Disorders ◽  
Healthy Subjects ◽  
Neospora Caninum ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Immunodeficiency Virus

ABSTRACT Considering that little is known about the epidemiology of Neospora caninum infection in humans, particularly in populations with high Toxoplasma gondii infection rates, the present study aimed to investigate the presence of antibodies to N. caninum in T. gondii-seropositive and -seronegative individuals. A total of 256 serum samples divided into four groups (61 samples from human immunodeficiency virus [HIV]-positive patients, 50 samples from patients with neurological disorders, 91 samples from newborns, and 54 samples from healthy subjects) were assessed for N. caninum and T. gondii serologies by indirect fluorescent-antibody test, enzyme-linked immunosorbent assay, and immunoblotting (IB). Immunoglobulin G antibodies to N. caninum were predominantly detected in HIV-infected patients (38%) and patients with neurological disorders (18%), while newborns and healthy subjects showed lower seropositivity rates (5% and 6%, respectively). Seropositivity to N. caninum was significantly associated with seropositivity to T. gondii in both HIV-infected patients and patients with neurological disorders. Seroreactivity to N. caninum was confirmed by IB, with positive sera predominantly recognizing the 29-kDa antigen of N. caninum. The results of this study indicate the presence of N. caninum infection or exposure in humans, particularly in HIV-infected patients or patients with neurological disorders, who could have opportunistic and concurrent infections with T. gondii. These findings may bring a new concern for the unstable clinical health of HIV-infected patients and the actual role of N. caninum infection in immunocompromised patients.

scholarly journals Seroprevalence of Toxoplasma gondii infection in domestic pigs reared under different management systems in Zimbabwe

2005 ◽  
Vol 72 (3) ◽  
Author(s):  
T. Hove ◽  
P. Lind ◽  
S. Mukaratirwa
Keyword(s):  
Toxoplasma Gondii ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Small Scale ◽  
Serum Samples ◽  
Serum Dilution ◽  
Domestic Pigs ◽  
Commercial Farms ◽  
Fattening Pigs

Serum samples from 474 domestic pigs (Sus scrofa) from Zimbabwe were tested for anti-Toxoplasma gondii IgG antibodies using the indirect fluorescent antibody test. The results showed that T. gondii infection is widespread in Zimbabwean pigs. Seroprevalence was lowest in fattening pigs from large and small-scale commercial farms that practise good hygiene (19.75 % of 238) and highest in backyard scavenging pigs (35.71 % of 70). Only 11.7 % (11) of the 127 positive samples had titres of > 1:400 and nine (81.82 %) of these 11 originated from pigs reared under poor hygienic conditions. A prevalence of 3.51 % was found in the same group of fattening pigs using an indirect IgG enzyme-linked immunosorbent assay at the single serum dilution of 1:400. The serosurvey shows the importance of modern intensive husbandry systems in reducing the prevalences of T. gondii infection in domestic pigs.

scholarly journals Diagnostic Performance of Assays for the Detection of Anti-Porcine Reproductive and Respiratory Syndrome Virus Antibodies in Serum and Muscle Transudate (“Meat Juice”) Based on Samples Collected under Experimental Conditions

2008 ◽  
Vol 20 (6) ◽  
pp. 735-743 ◽  
Author(s):  
Ramon M. Molina ◽  
Wayne Chittick ◽  
Eric A. Nelson ◽  
Jane Christopher-Hennings ◽  
Raymond R. R. Rowland ◽  
...  
Keyword(s):  
Neutralizing Antibodies ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Neutralization Assay ◽  
Respiratory Syndrome Virus ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Testing ◽  
Serum Samples ◽  
Experimental Conditions ◽  
Meat Juice

Three assays were evaluated for their ability to detect antibodies against Porcine reproductive and respiratory syndrome virus (PRRSV) in porcine muscle transudate (“meat juice”) samples. Samples were derived from 91 pigs inoculated with PRRSV isolate VR-2332 and 46 age-matched controls. Serum and muscle ( Musculus longissimus dorsi) samples were collected from randomly selected animals euthanized at ∼14-day intervals from 28 to 202 days postinoculation. Serum samples were assayed at a dilution of 1:40, and muscle transudate samples were assayed at 5 dilutions (1:2, 1:5, 1:10, 1:20, 1:40) using a commercial PRRSV antibody enzyme-linked immunosorbent assay (ELISA). In addition, muscle transudate samples were tested using an indirect fluorescent antibody test (IFAT) at 5 dilutions (1:2, 1:5, 1:10, 1:20, 1:40). Attempts to assay muscle transudate samples for neutralizing antibodies using a modified fluorescent focus neutralization assay were unsuccessful. Receiver operator characteristic (ROC) curve analyses were used to estimate cutoff thresholds and the associated diagnostic sensitivities and specificities for ELISA and IFAT at each dilution. For ELISA, muscle transudate samples at the ROC-optimized cutoffs were >95% sensitive and 100% specific at each dilution. At a cutoff dilution of ≥1:5, the IFAT diagnostic sensitivity and specificity of muscle transudate was estimated at 63.3% and 100%, respectively. These findings validated the use of muscle transudate samples in PRRSV surveillance programs based on ELISA antibody testing.

scholarly journals Serologic evidence of equine granulocytic anaplasmosis in horses from central West Brazil

2010 ◽  
Vol 19 (3) ◽  
pp. 135-140 ◽  
Author(s):  
Carlos Augusto Salvagni ◽  
Ana Sílvia Dagnone ◽  
Tiago Salles Gomes ◽  
Jozivaldo Silva Mota ◽  
Gisele Maria Andrade ◽  
...  
Keyword(s):  
Fluorescent Antibody ◽  
Antibody Test ◽  
Buffy Coat ◽  
Molecular Techniques ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Theileria Equi ◽  
Nested Polymerase Chain Reaction ◽  
Granulocytic Ehrlichiosis ◽  
Granulocytic Anaplasmosis

Ehrlichiosis is a zoonotic disease caused by gram-negative and intracellular obligatory bacterial organisms. Equine Granulocytic Anaplasmosis - EGA (formerly Equine Granulocytic Ehrlichiosis, EGE) is a seasonal disease, normally self-limited in horses. There are few reports in Brazil about this ehrlichial agent, as well as its natural vectors. Nowadays, veterinarians are considering the suspicion of EGA in horses with suggestive symptoms of ehrlichiosis and which do not respond to piroplasmosis treatment. The aim of the present study was to identify horses exposed to the agent A. phagocytophilum by serological and molecular techniques. Twenty equine blood and serum samples from the central West region of Brazil were evaluated by microscopic examination of buffy coat smear, enzyme-linked immunosorbent assay (ELISA), indirect fluorescent antibody test (IFAT) and nested polymerase chain reaction (nPCR). Additionally, the serodiagnosis of Theileria equi by IFA and ELISA were carried out, as well as molecular diagnosis by nPCR. Thirteen (65%) serum samples were positive for A. phagocytophilum by ELISA, but none of them were positive by buffy-coat smear examination or nPCR. Antibodies IgG anti-T. equi were detected in 18 (90%) and 17 (85%) horses by IFA and ELISA, respectively and the agent was detected in 9 (45%) animals by nPCR. Our data may be considered as important information to understanding the occurrence of EGA and equine piroplasmosis in central West Brazil.

scholarly journals Cloning of a Novel Babesia equi Gene Encoding a 158-Kilodalton Protein Useful for Serological Diagnosis

2005 ◽  
Vol 12 (2) ◽  
pp. 334-338 ◽  
Author(s):  
Haruyuki Hirata ◽  
Naoaki Yokoyama ◽  
Xuenan Xuan ◽  
Kozo Fujisaki ◽  
Naoyoshi Suzuki ◽  
...  
Keyword(s):  
Gene Product ◽  
Fluorescent Antibody ◽  
Horse Serum ◽  
Antibody Test ◽  
Immune Serum ◽  
Phage Library ◽  
Enzyme Linked Immunosorbent Assay ◽  
Infected Horse ◽  
Reading Frame ◽  
Babesia Equi

ABSTRACT In this study, we characterized a Babesia equi Be158 gene obtained by immunoscreening a B. equi cDNA expression phage library with B. equi-infected horse serum. The Be158 gene consists of an open reading frame of 3,510 nucleotides. The recombinant Be158 gene product was produced in Escherichia coli and used for the immunization of mice. In Western blot analysis, mouse immune serum against the Be158 gene product recognized 75- and 158-kDa proteins from the lysate of B. equi-infected erythrocytes. In an indirect fluorescent-antibody test with the mouse immune serum, the Be158 antigen appeared in the cytoplasm of Maltese cross-forming parasites (which consist of four merozoites) and was located mainly in the extraerythrocytic merozoite body. When the recombinant Be158 gene product was used in an enzyme-linked immunosorbent assay as a serological antigen, it was found to react to B. equi-infected horse sera, indicating that the Be158 gene product is useful as a serologically diagnostic antigen for B. equi infection.

scholarly journals Seroepidemiological aspects ofLeishmania spp. in dogs in the Itaguai micro-region, Rio de Janeiro, Brazil

2013 ◽  
Vol 22 (1) ◽  
pp. 39-45 ◽  
Author(s):  
Claudia Bezerra da Silva ◽  
Joice Aparecida Rezende Vilela ◽  
Marcus Sandes Pires ◽  
Huarrisson Azevedo Santos ◽  
Aline Falqueto ◽  
...  
Keyword(s):  
Rural Areas ◽  
Rio De Janeiro ◽  
Fluorescent Antibody ◽  
Antibody Test ◽  
Canine Leishmaniasis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Factors Associated ◽  
Definition Of ◽  
Forest Streams

This study evaluated factors associated with the frequency ofLeishmania spp. antibodies in dogs residing in the Itaguai micro-region, State of Rio de Janeiro, Brazil. Blood samples were collected from 524 dogs. The serum samples were submitted to indirect fluorescent antibody test (IFAT) and enzyme-linked immunosorbent assay (ELISA) forLeishmania spp. The frequency of seropositive dogs was 28.24% (n = 148) in the micro-region, and among the three municipalities within that region, the highest frequency (p < 0.05) was observed in Seropedica (59.46%), followed by Itaguai (29.05%) and Mangaratiba (11.49%). Regarding factors associated with the host, mongrel dogs and those over the age of two presented higher frequency of antibodies to Leishmaniaspp. (p < 0.05). Concerning factors related to the environment and habits of the animal, dogs residing in rural areas (FR = 1.67, p = 0.0002), living outside the residence (FR = 1.42, p = 0.0197), with access to forest, streams and pastures (FR = 2.81, p = 0.0007), remaining loose (FR = 1.66, p = 0.0073), and those that had no shelter (FR = 2.16, p < 0.0001) were more likely to be seropositive. Canine leishmaniasis is a disease with high occurrence in the Itaguai micro-region, and aspects such as the definition of breed, age, habits and care by owners showed significant association in this micro-region.

scholarly journals Antibodies anti-trypanosomatides in domestic cats in Paraná: who is at highest risk of infection?

2018 ◽  
Vol 27 (2) ◽  
pp. 232-236 ◽  
Author(s):  
Andressa Maria Rorato Nascimento de Matos ◽  
Eloiza Teles Caldart ◽  
Fernanda Pinto Ferreira ◽  
Keila Clarine Monteiro ◽  
Marielen de Souza ◽  
...  
Keyword(s):  
Antibody Test ◽  
Enzyme Linked Immunosorbent Assay ◽  
Domestic Cats ◽  
Serum Samples ◽  
Animal Group ◽  
Animal Protection ◽  
Leishmania Spp ◽  
In Series ◽  
Different Populations ◽  
Simple Logistic

Abstract The aim of this study were to detect antibodies anti-Leishmania spp. and anti-Trypanosoma cruzi in two different populations of domestic cats (Felis catus domesticus) from North Paraná referred for surgical castration and to determine which characteristics of the animals studied may be associated with seropositivity. Serum samples from 679 cats were analyzed using enzyme-linked immunosorbent assay (ELISA) and indirect immunofluorescence antibody test (IFAT) in series. Associations between age, sex, race, year of care and animal group were verified using the simple logistic regression. Percentage of 8.5% (58/679) of cats were positive for Leishmania spp. and 7.6% (51/673) for T. cruzi by the tests ELISA and IFAT. Animals collected by non-governmental animal protection organizations presented more seropositivity for Leishmania spp. (p<0.0001). Results shown that Leishmania spp. and T. cruzi are present in domestic cats in the northern part of the state of Paraná, as well as, owners of non-governmental animal protection organizations may be more exposed to leishmaniasis when compared to other animal owners evaluated in the present study.

scholarly journals Standardization of dot-ELISA for the serological diagnosis of toxocariasis and comparision of the assay with ELISA

1992 ◽  
Vol 34 (1) ◽  
pp. 55-60 ◽  
Author(s):  
Eide Dias Camargo ◽  
Paulo Mutuko Nakamura ◽  
Adelaide José Vaz ◽  
Marcos Vinícius da Silva ◽  
Pedro Paulo Chieffi ◽  
...  
Keyword(s):  
Low Cost ◽  
Clinical Signs ◽  
Toxocara Canis ◽  
Enzyme Linked Immunosorbent Assay ◽  
Serum Samples ◽  
Specific Antibodies ◽  
Normal Individuals ◽  
Before And After ◽  
Dot Elisa ◽  
Stability Time

The dot-enzyme-linked immunosorbent assay (dot-ELISA) was standardized using somatic (S) and excretory-secretory (ES) antigens of Toxocara-canis for the detection of specific antibodies in 22 serum samples from children aged 1 to 15 years, with clinical signs of toxocariasis. Fourteen serum samples from apparently normal individuals and 28 sera from patients with other pathologies were used as controls. All samples were used before and after absorption with Ascaris suum extract. When the results were evaluated in comparison with ELISA, the two tests were found to have similar sensitivity, but dot-ELISA was found to be more specific in the presence of the two antigens studied. Dot-ELISA proved to be effective for the diagnosis of human toxocariasis, presenting advantages in terms of yield, stability, time and ease of execution and low cost.

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