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 Serodiagnosis of Human Granulocytic Ehrlichiosis by a Recombinant HGE-44-Based Enzyme-Linked Immunosorbent Assay

1999 ◽  
Vol 37 (11) ◽  
pp. 3540-3544 ◽  
Author(s):  
Jacob W. Ijdo ◽  
Caiyun Wu ◽  
Louis A. Magnarelli ◽  
Erol Fikrig
Keyword(s):  
Immunosorbent Assay ◽  
Fluorescent Antibody ◽  
Immunoblot Analysis ◽  
Normal Serum ◽  
Enzyme Linked Immunosorbent Assay ◽  
Antibody Testing ◽  
Serum Samples ◽  
Granulocytic Ehrlichiosis ◽  
Healthy Humans ◽  
Human Granulocytic Ehrlichiosis

Current antibody testing for human granulocytic ehrlichiosis relies predominantly on indirect fluorescent-antibody assays and immunoblot analysis. Shortcomings of these techniques include high cost and variability of test results associated with the use of different strains of antigens derived from either horses or cultured HL-60 cells. We used recombinant protein HGE-44, expressed and purified as a maltose-binding protein (MBP) fusion peptide, as an antigen in a polyvalent enzyme-linked immunosorbent assay (ELISA). Fifty-five normal serum samples from healthy humans served as a reference to establish cutoff levels. Thirty-three of 38 HGE patient serum samples (87%), previously confirmed by positive whole-cell immunoblotting, reacted positively in the recombinant ELISA. In specificity analyses, serum samples from patients with Lyme disease, syphilis, rheumatoid arthritis, and human monocytic ehrlichiosis (HME) did not react with HGE-44–MBP antigen, except for one sample (specificity, 98%). We conclude that recombinant HGE-44 antigen is a suitable antigen in an ELISA for the laboratory diagnosis and epidemiological study of HGE.

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 Occurrence of Theileria equi in horses raised in the Jaboticabal microregion, São Paulo State, Brazil

2010 ◽  
Vol 19 (4) ◽  
pp. 228-232 ◽  
Author(s):  
Cristiane Divan Baldani ◽  
Andrea Cristina Higa Nakaghi ◽  
Rosangela Zacarias Machado
Keyword(s):  
Blood Smear ◽  
Fluorescent Antibody ◽  
Sao Paulo ◽  
São Paulo ◽  
Serum Samples ◽  
Theileria Equi ◽  
Nested Polymerase Chain Reaction ◽  
Paulo State ◽  
Blood Smears ◽  
São Paulo State

Blood and serum samples from 170 horses raised in the Jaboticabal microregion, São Paulo State, Brazil, were collected and tested by microscopic examination of blood smears, indirect fluorescent antibody test (IFAT) and nested polymerase chain reaction (nPCR) for Theileria equi infections. The association among the test results was verified by the McNemar test. During the examination of thin blood smears, parasites were detected in six (3.52%) horses. Anti-T. equi antibodies were detected in 100% sera samples, with titers ranging between 1:80 and 1:5120. The nPCR based on the T. equi merozoite antigen gene (EMA-1) allowed the visualization of specie-specific amplified product in 108 (63.53%) horses. All six samples judged positive microscopically were also positive for nPCR. Statistical analysis indicated general disagreement (p < 0.0001) between IFAT and nPCR; IFAT and blood smear; and nPCR and blood smear on the detection of parasite carriers. The results of the present study indicate that T. equi is widely spread among horses in the Jaboticabal microregion, Northeast region of São Paulo State, Brazil.

scholarly journals Factors associated to Theileria equi in equids of two microregions from Rio de Janeiro, Brazil

2011 ◽  
Vol 20 (3) ◽  
pp. 235-241 ◽  
Author(s):  
Tiago Marques dos Santos ◽  
Erica Cristina Rocha Roier ◽  
Huarrisson Azevedo Santos ◽  
Marcus Sandes Pires ◽  
Joice Aparecida Rezende Vilela ◽  
...  
Keyword(s):  
Rio De Janeiro ◽  
Fluorescent Antibody ◽  
Prevalence Ratio ◽  
Antibody Test ◽  
Geographic Area ◽  
Southeastern Brazil ◽  
Amblyomma Cajennense ◽  
Serum Samples ◽  
Theileria Equi ◽  
Factors Associated

Serum samples from 714 equids of Itaguaí and Serrana microregions, Rio de Janeiro, southeastern Brazil, were examined by indirect fluorescent antibody test (titer 1:80) for Theileria equi. The prevalence in the microregions and factors associated with seropositivity were evaluated and the prevalence ratio (PR) calculated. The overall prevalence of T. equi infection was 81.09% (n = 579), with higher prevalence (p < 0.05) in the Itaguaí (85.43%) when compared to Serrana microregion (76.92%). The geographic area, altitude, farming condition and area of origin of equids were associated (p < 0.05) with seropositivity for T. equi. Equids reared in the Itaguaí microregion (PR = 1.11, p = 0.003) and at altitudes below 500 m (PR = 1.10; p = 0,014) were more likely to be seropositive for T. equi. Furthermore, when equids were born in the farm (PR = 1.10, p = 0.008) and reared with poor farming conditions (PR = 1.13, p = 0.018) they were more likely to be exposed to T. equi. The main ticks found on equids were Amblyomma cajennense and Dermacentor (Anocentor) nitens. The microregions studied are endemic areas for equine theileriosis and there exists enzootic stability for T. equi. Only factors related to the collection area of serum samples influenced the seropositivity of equids for T. equi in that region.

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 Infection with Agents of Human Granulocytic Ehrlichiosis, Lyme Disease, and Babesiosis in Wild White-Footed Mice (Peromyscus leucopus) in Connecticut

1999 ◽  
Vol 37 (9) ◽  
pp. 2887-2892 ◽  
Author(s):  
Kirby C. Stafford ◽  
Robert F. Massung ◽  
Louis A. Magnarelli ◽  
Jacob W. Ijdo ◽  
John F. Anderson
Keyword(s):  
Ixodes Scapularis ◽  
Peromyscus Leucopus ◽  
Fluorescent Antibody ◽  
Babesia Microti ◽  
Enzyme Linked Immunosorbent Assay ◽  
Rrna Gene ◽  
Serum Samples ◽  
Granulocytic Ehrlichiosis ◽  
Human Granulocytic Ehrlichiosis ◽  
Staining Methods

White-footed mice, Peromyscus leucopus, were captured in southern Connecticut during 1997 and 1998 to determine the prevalence of infections caused by granulocytic Ehrlichiasp., Borrelia burgdorferi, and Babesia microti. Of the 50 mice captured and recaptured, 25 of 47 (53.2%) and 23 of 48 (47.9%) contained antibodies to the BDS or NCH-1 Ehrlichiastrains, respectively, as determined by indirect fluorescent antibody (IFA) staining methods. The majority (83.3%) of 48 mice also contained antibodies to B. burgdorferi, as determined by enzyme-linked immunosorbent assay. Moreover, 20 of 26 (76.9%) contained antibodies to B. microti by IFA staining methods. In nested PCR tests using the 16S rRNA gene, the DNA of the human granulocytic ehrlichiosis (HGE) agent was detected in 17 of 47 mice (36.2%), but only 4 (23.5%) of these 17 mice were PCR positive at each capture. Antibody-positive reactions to granulocyticEhrlichia sp. organisms were detected in 17 of 23 (73.9%) of the PCR-positive mice. The sequences from PCR products from nine positive blood samples were identical to the HGE agent.Ehrlichia spp. were cultured from three of five mice captured in April 1998, including one that was PCR positive in April 1997. In addition, 2 of 14 larval Ixodes scapularis pools, which were attached to two PCR-positive mice, contained DNA of the HGE agent. A high percentage of white-footed mice are infected or have been infected naturally by the HGE agent with low-level persistent infection or frequent reinfection in some individual mice. However, the changes noted in the presence of DNA and antibodies in repeated blood and serum samples from individual mice over several months of field collection suggests that infection with granulocytic Ehrlichia is transient in most wild P. leucopus.

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 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.

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