Indirect Fluorescent Antibody Test Research Articles

Profiling of Mycobacterium avium subspecies paratuberculosis in the milk of lactating goats using antigen-antibody based assays

Johne's disease, caused by Mycobacterium avium subspecies paratuberculosis is endemic in the domestic livestock population, still it is not priority for control in the country. First time we used 'multiple assays’ for screening raw milk of 465 goats (farm/farmer's herds) to estimate bio-load and bio-type profile of bacilli. Each sample was screened by six tests and compared their sensitivity and specificity. Of 465 raw milk samples screened, bio-load of bacilli was 65.3% by six assays. Assay-wise bio-load was 49.4 and 62.7% in antigen and antibody detection tests, respectively. Bio-load was 48.8, 46.6, and 13.9% in Indirect Fluorescent Antibody Test (i_FAT), microscopy and IS900 PCR and 39.1, 57.4 and 55.6% in Indirect Enzyme Linked Immuno Sorbant Assay (i_ELISA), Dot Enzyme Linked Immuno Sorbant Assay (d_ELISA) and Latex Agglutination Test (LAT), respectively. Dot-ELISA was most sensitive followed by LAT, i_FAT, microscopy and i_ELISA. Milk DNA samples positive in IS900 PCR on bio-typing using IS1311 PCR_ Restriction Enzyme Analysis (IS1311 PCR_REA) revealed, 72.3% (47/65) were 'Indian Bison Type'. Milk was easy to collect sample and first time we used 'whole milk' as 'test sample' without centrifugation. High bio-load of MAP in milk underlined need for urgent control of disease in lactating goatherds. Bacilli was important 'Milk born' infection and on the basis of sensitivity, specificity, resources and requirements, of the ‘six assays’ most appropriate assay/s (single or in combination) can be chosen for the screening and diagnosis of Johne’s disease in lactating goatherds using whole milk as sample.

Socioeconomic vulnerability associated to Toxoplasma gondii exposure in southern Brazil.

Human toxoplasmosis, a protozoonosis caused by Toxoplasma gondii, has been described as a worldwide foodborne disease with important public health impact. Despite infection has reportedly varied due to differences in alimentary, cultural and hygienic habits and geographic region, social vulnerability influence on toxoplasmosis distribution remains to be fully established. Accordingly, the present study has aimed to assess T. gondii seroprevalence and factors associated to social vulnerability for infection in households of Ivaiporã, southern Brazil, with 33.6% population making half minimum wage or less, ranked 1,055th in population (31,816 habitants), 1,406th in per capita income (U$ 211.80 per month) and 1,021st in HDI (0.764) out of 5,570 Brazilian cities. Serum samples and epidemiological questionnaires were obtained from citizen volunteers with official City Secretary of Health assistance in 2015 and 2016. In overall, serosurvey has revealed 526/715 (73.57%) positive samples for anti-T. gondii antibodies by Indirect Fluorescent Antibody Test. Logistic regression has shown a significant increase associated to adults (p = 0.021) and elderly (p = 0.014) people, illiterates (p = 0.025), unemployment (p <0.001) and lack of household water tank (p = 0.039). On the other hand, sex (male or female), living area (urban or rural), yard hygiene, meat ingestion, sand or land contact, owning pets (dog, cat or both) were not significant variables of positivity for anti-T. gondii antibodies in the surveyed population. Although no significant spatial cluster was found, high intensity areas of seropositive individuals were located in the Kernel map where the suburban neighborhoods are located. In conclusion, socioeconomic vulnerability determinants may be associated to Toxoplasma gondii exposure. The increased risk due to illiteracy, adult or elderly age, unemployment and lack of household water tank were confirmed by multivariate analysis and the influence of low family income for seropositivity by the spatial analysis.

Development and application of an indirect enzyme-linked immunosorbent assay using recombinant S1 for serological testing of porcine epidemic diarrhea virus.

Porcine epidemic diarrhea virus (PEDV) causes severe infectious diseases in all ages of swine and leads to serious economic losses. Serologic tests are widely accepted and used to detect anti-PEDV antibodies that could indicate PEDV infection or vaccination. In this study, PEDV recombinant S1 protein (rS1) was expressed with the Bac-to-Bac system and purified by nickel-affinity chromatography. An indirect enzyme-linked immunosorbent assay based on rS1 (rS1-ELISA) was then developed and optimized by checkerboard assays with serial dilutions of antigen and serum. Serum samples from 453 domestic pigs and 42 vaccinated pigs were analyzed by the indirect fluorescent antibody (IFA) test and rS1-ELISA. Taking IFA as a gold standard, rS1-ELISA produced a high sensitivity (90.7%) and specificity (94.6%) by a receiver operating characteristic (ROC) curve. In addition, ROC analysis also revealed that rS1-ELISA was consistent with IFA (area under the curve 0.9583 ± 0.0082). This rS1-ELISA was then applied to antibody detection in inactivated PEDV vaccinated pigs. The antibody could be detected 2-4 weeks after the first inoculation. These results indicated that the rS1-ELISA established in this study provides a promising and reliable tool for serologic detection of anti-PEDV IgG antibodies in infected or vaccinated pigs.

Vertical transmission of Neospora caninum in bovine fetuses from a slaughterhouse in Brazil.

Neospora caninum is considered as one of the main causes of reproductive failure in cattle. Vertical transmission is the main route of infection in the bovine host and plays an important role in maintaining the parasite in the herd. Molecular detection of N. caninum is important to determine the occurrence of the disease and to evaluate the genetic diversity of the parasite. The present study aimed at assessing the vertical transmission of N. caninum using molecular techniques to detect the parasite in tissue samples from bovine fetuses collected in a slaughterhouse in the state of São Paulo, Brazil. Seventy fetuses and 70 blood samples from pregnant cows were collected in a slaughtering line. Fresh samples of heart and brain tissue from fetuses were analyzed using molecular assays. Serum samples from fetuses and cows were subjected to an indirect fluorescent antibody test (IFAT) to detect antibodies against N. caninum. Nested PCR targeting the internal transcriber 1 (ITS1) region of the protozoan organism was used in the molecular testing. From the total of fetuses examined, 71.42% were positive for N. caninum by PCR. A higher number of heart samples (47.1%) were positive for the parasite using this technique. Antibodies against the protozoa were detected in 12.9% of serum samples of cows; 2.8% of fetuses were seropositive for this pathogen. Our results show that vertical transmission of N. caninum occurs in cattle from this region of Brazil, and that the use of different diagnostic techniques contributes to successful diagnosis of congenital transmission of the parasite in cattle.

Toxoplasma gondii in Captive Wild Felids of Mexico: Its Frequency and Capability to Eliminate Oocysts.

There is little information about Toxoplasma gondii in wild felids, even when these species have been associated with cases of toxoplasmosis in humans. In this study, samples of serum and whole blood were collected from 42 felids from 10 different species, in 4 Mexican zoos. Stool samples from 36 animals were also collected, corresponding to 82% of the felids included in the study. Stool samples were used for the search of oocysts by light field microscopy and PCR. Serum samples were analyzed by indirect immunoglobulin G (IgG) enzyme-linked immunosorbent assay (ELISA) and indirect fluorescent antibody test (IFAT). DNA samples were purified from whole blood and stool for the amplification of a fragment of the SAG1 gene of T. gondii by a nested PCR (nPCR). The seroprevalence of IgG anti-T. gondii-specific antibodies by means of the ELISA was 100% (42/42) and 52.4% (22/42) by IFAT. The titers obtained varied from 1:80 to 1:2560. DNA of T. gondii was detected in 9.5% (4/42) of the blood samples by using nPCR. No oocysts were observed in the stool samples analyzed by light field microscopy. However, the DNA of the parasite was identified in 14.3% (5/35) of the stool samples evaluated. These results indicate a high prevalence of T. gondii in the studied populations of wild felids in captivity, with evidence of parasitemia and elimination of few oocysts even in adult hosts.

Factors associated with the seroprevalence of Neospora caninum (Apicomplexa: Toxoplasmatinae) in sheep from the State of Sergipe, Brazil

The aim of this study was to determine the prevalence and factors associated with the occurrence of antibodies against Neospora caninum in sheep from Sergipe, northeastern Brazil. A total of 932 sheep serum samples from 54 properties in 19 municipalities from the State of Sergipe, Brazil were collected and assayed using an indirect fluorescent antibody test (IFAT) to assess antibodies against N. caninum. A cut-off point of 1:50 was adopted and results showed that 12.45% (116/932) of sheep were serum-reactive. Based on an unconditional logistic regression, the presence of dogs on the property was associated with protection (OR= 0.323), whereas the use of exchanged or borrowed breeding males was associated with infection (OR= 22.287). These results indicate that the occurrence of antibodies against N. caninum is endemic in the State municipalities

Is the cat an important reservoir host for visceral leishmaniasis? A systematic review with meta-analysis

In recent years feline leishmanial infections (FLI) have been studied more than ever before in various parts of the world. However, evidence-based knowledge on FLI has remained unavailable. The main objectives of this study were to investigate the status of felines infected by Leishmania spp. worldwide. Data were extracted from 10 available databases over the period of 1982 to 2017. Overall, 78 articles fulfilled the inclusion criteria and were used for data extraction in this systematic review. The overall FLI prevalence by both serological and molecular methods was estimated at 10% (95% CI: 8%-14%). In Italy, both the seroprevalence (24 %) and PCR prevalence (21 %) were found to be higher than in other countries. The most common diagnostic test used was the indirect fluorescent antibody test (38.5%). Studies on mixed-breed felines were more common than those on other breeds, while the most common parasite species was L. infantum (63%). Our findings suggest that cats act as primary and/or secondary reservoir hosts in the transmission of the Leishmania spp. to humans and also to dogs, by sandflies, at least in endemic foci. Moreover, available data confirm the enzootic stability situation of FLI in several countries including some in Europe.

Anti-Neospora caninum antibodies in beef cattle from the northern region of Paraná state, Brazil

ABSTRACT: The presence of anti-Neospora caninum antibodies in beef cattle slaughtered in the northern region of the state of Paraná, Brazil, was evaluated. A total of 401 blood samples were collected; 281 samples from the municipality of Rolândia and 120 from the municipality of Borrazópolis, between April 2015 and November 2016. Of the total samples, 289 were from females and 112 from males, aged one and a half to eight years. Indirect fluorescent antibody test (IFAT) was performed, using a cut-off of 1:100. Variables were tabulated for statistical analyses (Fisher’s exact test and chi-square tests, p≤0.05). The analysis showed that of the 401 samples, 37 were positive for N. caninum, indicating a prevalence of 9.2 %, and observed titers were 1:100 (16), 1:200 (14), and 1:400 (7). The variables sex, age, and location did not differ statistically (p&gt;0.05). Our results showed a sero-occurrence of N. caninum in cattle slaughtered in the northern region of the state of Paraná.

Diagnosis Laboratorium Malaria

Malaria is a mosquito-borne disease caused by a parasite. Rapid and accurate diagnosis of malaria is integral to the appropriate treatment of affected individuals and in preventing the further spread of infection in the community. Left untreated, they may develop severe complications and die. Clinical diagnosis is based on the patient’s symptoms and on physical findings at examination. People with malaria often experience fever, chills, and flu-like illness. Clinical findings should always be confirmed by a laboratory test for malaria. Microscopic examination remains the “gold standard” for laboratory confirmation of malaria. Species determination is made based on morphological characteristics of the four species of human malaria parasites and the infected red blood cells. However, many simian species are largely indistinguishable from the four human species by microscopy. Molecular techniques such as polymerase chain reaction (PCR) and microsatellite testing can assist in definitive species determination. An alternate way of establishing the diagnosis of malaria infection are by rapid diagnostic test (RDT), indirect fluorescent antibody (IFA) test and enzyme immunoassays (EIA).

Where, when, and how the diagnosis of human visceral leishmaniasis is defined: answers from the Brazilian control program.

BACKGROUND Timely diagnosis is recommended by the Brazilian Visceral Leishmaniasis (VL) Surveillance and Control Program to reduce case fatality. Attempts at assessing this topic in Brazil are scarce. OBJECTIVE This study aimed to describe where, when, and how the diagnosis of VL has been performed in a Brazilian endemic setting.METHODS Data of all autochthonous cases confirmed between 2011 and 2016 (N = 81) were recorded. The care-seeking itinerary until the confirmation of VL diagnosis was assessed among 57 patients.FINDINGS The majority of VL cases (79.1%) were reported by referral hospitals. The patients mainly sought primary health care centres at the onset of symptoms. However, they had to visit seven health services on average to achieve a confirmed diagnosis. The time from the onset of symptoms to the diagnosis of VL (TD) ranged from 1-212 (median, 25) days. The TD was longer among adult patients. There was a direct correlation between the patient’s age and TD (r = 0.22; p = 0.047) and a higher occurrence of deaths due to the disease among older patients (p = 0.002). Almost all the patients (98.9%) underwent laboratory investigation, and the VL diagnosis was mainly confirmed based on clinical-laboratory criteria (92.6%). Positive results for the indirect fluorescence antibody test (22.7%) and parasitological examination plus rk39-based immunochromatographic tests (21.3%) were commonly employed.MAIN CONCLUSIONS VL diagnosis was predominantly conducted in hospitals with a long TD and wide application of serology. These findings may support measures focused on early diagnosis, including a greater involvement of the primary health care system.

Besnoitia besnoiti seroprevalence in beef, dairy and bullfighting cattle in Catalonia (north-eastern Spain): A cross-sectional study

Besnoitia besnoiti is the causative agent of bovine besnoitiosis, a chronic and debilitating disease of cattle that recently re-emerged and seems to be spreading in Europe. A cross-sectional serological study was carried out in different cattle herds in Catalonia, north-eastern Spain, to determine the seroprevalence of B. besnoiti in the region. A total of 791 serum samples (beef cattle n = 338, dairy cattle n = 291; bullfighting cattle n = 162) were tested. Sera were first screened for antibodies against Besnoitia using a commercial Enzyme-Linked Immunosorbent Assay (ELISA) applying a cut-off that was lower than that recommended by the manufacturer in order to reach highest sensitivity. Sera above the chosen cut-off of 15% positivity (PP) were further tested by the Indirect Fluorescent Antibody Test (IFAT) and respectively positive results were confirmed by a B. besnoiti tachyzoite-based immunoblot. A total of 504/791 (63.7%) sera showed ELISA values above the selected cut-off, and 91 of these samples also yielded positive results in IFAT (cut-off titre 1:200). By immunoblot, a positive result was obtained in 93.4% (85 out of the 91) of the IFAT-positive samples. Interestingly, all confirmed Besnoitia-seropositive cases corresponded exclusively to beef cattle from the Pyrenees area, resulting in a prevalence of 25.1% (85/338) at the animal level and of 46% (36/78) at the herd level in this cattle group. No specific antibodies against Besnoitia could be detected in dairy and bullfighting cattle. The obtained results suggested that Besnoitia infections are present in Catalonia, consequently, diagnosis of this parasitic infection should be included in the sanitary control and before trading and movement of animals.

A Descriptive Study of Lectin Histochemistry of the Placenta in Cattle following Inoculation of Neospora caninum

The aim of this study was to describe the lectin-binding pattern in the placentas of cows infected experimentally with Neospora caninum. Four cows were inoculated intravenously with 1×108 tachyzoites of the NC-1 strain of N. caninum at 150±7 days of pregnancy. Two control cows were administered a placebo. An indirect fluorescence antibody test (IFAT) was performed on serum samples obtained before and after the inoculation. The cows were killed at 30 and 37 days post inoculation. Samples of placenta were taken for histopathology and lectin histochemistry. Fetal tissues and fluids were collected for histopathology and IFAT, respectively. All infected cows had high antibody titres. All fetuses had characteristic histopathological lesions, including non-suppurative meningoencephalitis, myocarditis, hepatitis and myositis, suggesting N. caninum infection. Only two infected fetuses developed specific antibodies. Mild non-suppurative inflammatory infiltrates were recorded in the placentae. Differences in the lectin-binding pattern were observed between infected animals and controls in the glycocalyx (CON-A and WGA) and apical cytoplasm (RCA-I and CON-A) of the trophoblastic cells; giant trophoblastic cells (CON-A and DBA); glycocalyx (PNA, WGA) and apical cytoplasm (CON-A, WGA, PNA, DBA and RCA-I) of endometrial cells; trophoblast of the interplacentomal region (WGA); endothelium (CON-A, SBA, RCA-1 and WGA); and finally, mesenchyme (CON-A, RCA-1, SBA, PNA and DBA). These findings indicate that there is a distinctive pattern of lectin binding in the placenta of cattle infected with N. caninum. The direct effect of the presence of the protozoa as well as the altered expression of cytokines could explain these changes in the maternofetal interface.

Serological diagnosis and risk factors for Coxiella burnetii in goats and sheep in a semi-arid region of Northeastern Brazil.

Coxiella burnetii is a zoonotic agent transmitted mainly by small ruminants. In Brazil the disease has been classified as a notifiable disease since 2013, when human cases were reported. This study aimed to identify risk factors associated with the presence of anti- Coxiella burnetii antibodies in goats and sheep in a semiarid region of Northeastern Brazil. Sera of 412 goats and 403 sheep from municipality of Petrolina, Pernambuco, were examined by the Indirect Fluorescent Antibody Test (IFAT) against antigens of C. burnetii. Information about management variables (independent variables) that could be associated with the presence of the microorganism (dependent variables) were obtained from the supervisor of each farm. It was determined that 2.2% (9/412) of the goats and 2.1% (9/403) of the sheep had antibodies reactive to C. burnetii. The presence of anti-C. burnetii antibodies was associated with the dry area of the Sequeiro (a region in the northern part of the municipality of Petrolina) (P = 0.025), male sheep (P = 0.020), and intensive goat breeding (P = 0.005). This study therefore showed the presence of anti-C. burnetii antibodies in goat and sheep, confirming for the first time that this agent is likely circulating among goat herds in the Caatinga Biome, semi-arid of Brazil.

TRANSMISSION DYNAMICS OF TOXOPLASMA GONDII IN ARCTIC FOXES (VULPES LAGOPUS): A LONG-TERM MARK-RECAPTURE SEROLOGIC STUDY AT KARRAK LAKE, NUNAVUT, CANADA.

Transmission dynamics of Toxoplasma gondii, a parasite of importance for wildlife and human health, are enigmatic in the Arctic tundra, where free-ranging wild and domestic felid definitive hosts are absent and rarely observed, respectively. Through a multiyear mark-recapture study (2011-17), serosurveillance was conducted to investigate transmission of T. gondii in Arctic foxes (Vulpes lagopus) in the Karrak Lake region, Nunavut, Canada. Sera from adult foxes and fox pups were tested for antibodies to T. gondii by using serologic methods, including the indirect fluorescent antibody test, direct agglutination test, and modified agglutination test. The overall seroprevalence was 39% in adults and 17% in pups. Mature foxes were more likely to be exposed (seroconvert) than young foxes (less than 1 yr old), with the highest level of seroprevalence in midaged foxes (2-4 yr old). Pups in two different litters were seropositive on emergence from the den, around 5 wk old, which could have been due to passive transfer of maternal antibody or vertical transmission of T. gondii from mother to offspring. The seropositive pups were born of seropositive mothers that were also seropositive the year before they gave birth, suggesting that vertical transmission might not be limited to litters from mothers exposed to T. gondii for the first time in pregnancy. All recaptured seropositive foxes remained seropositive on subsequent captures, suggesting that antibodies persist or foxes are constantly reexposed or a combination of both. The results of this study provided insights into how foxes were likely exposed to T. gondii, the dynamics of antibody persistence and immune response, and how the parasite was maintained in a terrestrial Arctic ecosystem in the absence of felid definitive hosts.

An ELISA-based method for detection of rabies virus nucleoprotein-specific antibodies in human antemortem samples.

Rabies is a fatal encephalitic disease in humans and animals caused by lyssaviruses, most commonly rabies virus (RABV). Human antemortem diagnosis of rabies is a complex process involving multiple sample types and tests for the detection of antibodies, antigen (protein), and nucleic acids (genomic RNA). Serological diagnosis of human rabies includes the detection of either neutralizing or binding antibodies in the cerebrospinal fluid (CSF) or serum samples from unimmunized individuals without prior rabies vaccination or passive immunization with purified immunoglobulins. While neutralizing antibodies are targeted against the surface-expressed glycoprotein (G protein), binding antibodies to viral antigens are predominantly against the nucleoprotein (N protein), although there can be antibodies against all RABV-expressed proteins. To determine N protein-specific antibody responses in the CSF and serum during RABV infection, we developed an enzyme-linked immunosorbent assay (ELISA) with purified recombinant N protein expressed in E. coli. N protein-specific immunoglobulin (Ig) subtypes IgG and IgM were detected in the CSF or serum of previously diagnosed human rabies cases. In addition, anti-N protein seroconversion was demonstrated over the course of illness in individual rabies cases. We compared the N protein ELISA results to those of an indirect fluorescent antibody (IFA) test, the current binding antibody assay used in diagnosis, and show that our ELISA is consistent with the IFA test. Sensitivity and specificity of the N protein ELISA ranged from 78.38–100% and 75.76–96.77% with respect to the IFA results. Our data provide evidence for the use of an N protein ELISA as an additional option for the detection of RABV-specific IgG or IgM antibodies in human CSF or serum specimens.

Prevalence of visceral leishmaniasis in A population of free-roaming dogs as determined by multiple sampling efforts: A longitudinal study analyzing the effectiveness of euthanasia

Free-roaming dogs affected by visceral leishmaniasis (VL) contribute to the geographical expansion of the disease and require special attention from health authorities. The objectives of the present study were to determine the prevalences of VL in a population of free-roaming dogs in an endemic region of Brazil, to establish the spatial distribution of infected dogs, and to examine the effectiveness of euthanasia of infected dogs in controlling the disease in this particular population. Dogs were captured every two months during seven sampling efforts. Capture locations were georeferenced and captured dogs were assessed for the presence of anti-Leishmania antibodies using an enzyme-linked immunosorbent assay (ELISA) as a screening test and the indirect fluorescent antibody test (IFAT) as the confirmatory procedure. Dogs that were seropositive by both assays were considered infected and were submitted to immediate euthanasia. After the end of the collection period, stored sera were evaluated with the Dual-Path Platform test (DPP). Animals positive by this method and by ELISA were also considered infected as currently recommended by the Brazilian Ministry of Health. Spatial analysis was performed using the Kernel technique. A total of 328 dogs were captured at least once during the sampling period, 25 (7.6%) of them were seropositive by ELISA and IFAT and 27 (8.2%) by DPP and ELISA. The prevalence of VL showed an overall decreasing trend. However, even with periodical euthanasia, it was not possible to eliminate the infection and increased prevalences were observed in the fourth and seventh samplings. There was a high overall agreement between the two criteria for defining infection. None of the dogs that tested negative by IFAT at the first capture seroconverted in the subsequent captures but a number of dogs exhibited changes in serological status over time. From the three dogs initially tested negative by ELISA and IFAT, but tested positive by the protocol currently adopted in Brazil, two became negative in subsequent recaptures. Spatial analysis revealed that infected animals concentrated in areas with a high density of free-roaming dogs. The existence of VL among homeless dogs may contribute significantly in the persistence of the disease among the human population, despite the practice of periodical euthanasia. The operational and ethical implications associated with euthanasia of free-roaming dogs, and the failure to control the transmission of VL among this particular population, led us to conclude that interventions promoting responsible ownership of pets may be a more effective strategy.

Serologic Survey of Selected Viral Pathogens in Free-Ranging Eurasian Brown Bears ( Ursus arctos arctos) from Slovakia.

We tested sera of 24 free-ranging European brown bears ( Ursus arctos) from six regions of Slovakia for antibodies to 10 viral agents. We tested sera by an indirect fluorescence antibody test for antibodies to canine distemper virus (CDV), canine coronavirus (CCV), canine parvovirus type 2 (CPV-2), canine adenovirus, canine parainfluenza virus type 2 (CPIV-2), and canine herpesvirus type 1 (CHV-1). We used an enzyme-linked immunosorbent assay for detection of antibodies to hepatitis E virus, bluetongue virus, West Nile virus (WNV), and Aujeszky's disease virus (ADV). We detected antibodies to CDV, CHV-1, CPV-2, CPIV-2, CCV, WNV, and ADV in seven (29%), three (12%), two (8%), two (8%), one (4%), one (4%), and one (4%) bear, respectively. Evidence of exposure of free-ranging European brown bears to CCV and ADV has not been reported.

Prevalence of antibodies against Toxoplasma gondii and Neospora spp. in equids of Western Pará, Brazil

The present study aimed to determine the prevalence of antibodies against Toxoplasma gondii and Neospora spp. in equids raised for distinct purposes in the western state of Pará, Brazil, and to identify potential risk factors associated with parasite infection. A cross-sectional study was conducted with cluster sampling in 101 farms from 18 municipalities (farm horses). In visited municipalities, samples from sport and cart horses were included. Serum was obtained for detecting antibodies against T. gondii and Neospora spp. using indirect fluorescent antibody test, with a cut-off point of 1:64 and 1:50, respectively. Association analysis, using the chi-square test, was performed to evaluate possible risk factors related to the prevalence of antibodies. A total of 1298 equids were sampled (947 farm, 240 sport, and 111 cart horses), including 1244 horses, 2 donkeys, and 52 mules. The number of equids sampled per property ranged from 1 to 49. Of the total examined, 134 (10.3%) had antibodies against T. gondii, and of the 18 municipalities visited, 11 (61%) had at least one positive animal. Seventy-three of the 1298 equids (5.62%) from 44% of the municipalities tested positive for Neospora spp. The type of the animal (farm, sport, or carthorse) was not associated with the prevalence of antibodies against T. gondii; however, antibodies against Neospora spp. was more prevalent in carthorses (P = 0.001) (13.5%) than in farm (5.0%) and sport (4.6%) horses. Breed of the horse and the presence of cats were risk factors for the prevalence of antibodies against T. gondii, and presence of dogs was a risk factor for the prevalence of antibodies against Neospora spp. This is the first report of seropositive equids for T. gondii and Neospora spp. in the state of Pará contributing to the knowledge on epidemiology of the two parasites.

Prevalence and risk factors associated with Leishmania spp. and Trypanosoma cruzi infections in dogs presented at veterinary clinics in João Pessoa, Paraíba state, northeastern Brazil

This study aimed to determine the prevalence and risk factors associated with Leishmania spp. and Trypanosoma cruzi infections in dogs presented at veterinary clinics in the city of João Pessoa, Paraíba state, northeastern Brazil. A total of 384 dogs from 34 veterinary clinics were sampled between April 2015 and May 2016. For serological diagnosis of canine visceral leishmaniasis and Chagas disease, the indirect fluorescent antibody test was used. Reactive sera were submitted to a confirmatory test using the S7 ELISA. The seroprevalence of Leishmania spp. was 2.8% (11/384), and that of T. cruzi was 1.5% (6/384). Two animals were positive for both parasites. The variable “access to the street” was identified as a risk factor for Leishmania spp. infection (OR = 4.81; 95% CI = 1.23–18.75). We concluded that a significant proportion of dogs presented at veterinary clinics in João Pessoa are seropositive for Leishmania spp. or T. cruzi infections, and we suggest that greater care be taken with dogs that have access to the street.

NEOSPORA CANINUM-SPECIFIC ANTIBODIES IN FREE-RANGING WHITE-LIPPED PECCARIES ( TAYASSU PECARI) FROM THE PERUVIAN AMAZON: DETECTION OF ANTIBODIES IN SERUM AND EVALUATION OF INDIRECT FLUORESCENT ANTIBODY TEST WITH HETEROLOGOUS SECONDARY ANTIBODY.

Neospora caninum is a coccidian parasite originally reported in dogs and widely prevalent in numerous species of wild and domestic animals and has as definitive hosts some species of canids. The white-lipped peccary (WLP) ( Tayassu pecari) is a Tayassuidae mammal, found from Mexico to south of Brazil and north of Argentina. It is a game species with great economic importance in the Peruvian Amazon. Blood samples from 101 WLPs were collected from near or within three different conservation reserves located in the southeastern region of the Peruvian Amazon. For the detection of antibodies against N. caninum, indirect fluorescent antibody tests (IFAT) were performed using collared peccary ( Pecari tajacu) and swine ( Sus scrofa domesticus) heterologous secondary antibodies. For both IFAT tests, the cutoff was 1:50. Positive samples were titrated by a two fold serial dilution. In addition to IFAT, samples were also analyzed using an immunoblotting test (IB) with anti-swine conjugate. To confirm the viability of the anti-swine conjugate, the results of these samples previously tested by a modified agglutination test (MAT) for Toxoplasma gondii were used as reference. From the total of 101 samples tested, 5 (4.9%) were N. caninum positive by the three tests and an extra sample was positive by both IFATs and negative in the IB. Comparing both IFATs and considering IB as the gold standard, the relative sensitivity of IFATs was 100%, the specificity was 98.9%, the positive predictive value was 83.3%, and the negative predictive value was 100%. The agreement between tests was characterized by a κ value of 0.904 (95% confidence interval, 0.717 to 1.0) and an SE of 0.095. This is the first report of N. caninum antibodies in free-ranging T. pecari, and swine and collared peccary conjugate can be used as a secondary antibody for detection of antibodies in Tayassu species.