First report of seroprevalence of Toxoplasma gondii and Neospora caninum in dairy sheep from Humid Pampa, Argentina

The aim of this study was to describe the occurrence of antibodies to Toxoplasma gondii and Neospora caninum in dairy sheep from the Humid Pampa region, Argentina. Blood samples from 704 dairy sheep belonging to six flocks were collected. Using a cut off titer of 1:50, an indirect fluorescence antibody test was used. Antibodies to T. gondii or N. caninum were detected in 17.3 % (n?=?122) and 3 % (n?=?21), respectively. All the flocks had at least one seropositive animal to T. gondii but two of them had no seropositive sheep to N. caninum. Fifty-two of 122 (42.6 %) positive samples to T. gondii had antibody titers higher than 1:400. There was a significantly higher proportion of T. gondii seropositive animals in females and older sheep (p?<?0.05). Ten of 21 (52.3 %) positive samples to N. caninum had antibody titers higher than 1:400. This is the first report of seroprevalence of T. gondii and N. caninum in dairy sheep from Humid Pampa, Argentina. Further research is required for a better understanding of the role of toxoplasmosis and neosporosis in dairy sheep in Argentina.     

Prevalence of anti-Toxoplasma gondii and anti-Neospora caninum antibodies in sheep from Mossoró, Rio Grande do Norte, Brazil

Toxoplasma gondii is an obligate intracellular parasite with a variety of hosts, responsible for reproductive problems and economic losses in sheep flocks. Neospora caninum was recently identified and its clinical presentation in sheep is similar to that of toxoplasmosis, which can cause repeated abortions, though less frequently in this species. In order to confirm the prevalence of these agents in the city of Mossoró, Rio Grande do Norte, Brazil, 409 serum samples from adult sheep (364 females and 45 males) were tested by the indirect immunofluorescence antibody test, using cut-off point at a dilution of 1:64 and 1:50 for T. gondii and N. caninum, respectively. From the 35 properties examined, 23 (65.7%) had at least one seropositive animal for T. gondii and six (17.1%) for N. caninum. The prevalence of seropositive animals for T. gondii was 20.7% and for N. caninum 1.8%. There was no association between the presence of the agent’s antibody and gender, reports of reproductive problems and presence of dogs and/or cats in the properties. T. gondii is well distributed and N. caninum has low prevalence in sheep and in the properties of the studied region.     

Prevalence of anti-<Emphasis Type="Italic">Toxoplasma gondii</Emphasis> antibodies in sport horses from Qazvin,Iran

In the present study, the prevalence of antibodies to Toxoplasma gondii in sport horses of Qazvin was examined using modified agglutination test (MAT). On 52 horse sera totally examined for anti-Toxoplasma antibodies, 37 horses (71.2%) were seropositive by MAT. Results of the present study showed a high rate of Toxoplasma infection in horses in Qazvin area. More comprehensive study on equine toxoplasmosis is recommended.     

Serological Survey of Toxoplasma gondii Infection in the Domestic Goose (Anser domestica) in Southern China

In the present study, the antibodies to Toxoplasma gondii in 191 farm‐bred and 83 house‐bred geese (Anser domestica) were assessed for the prevalence of T. gondii infection in southern China with the modified agglutination test. Antibodies to T. gondii (MAT ≥ 1 : 5) were found in 27 (14.14%) of farm‐bred geese and 14 (16.87%) of house‐bred geese. Geese infected with T. gondii may be a source of T. gondii infection for humans and cats.     

Comparison of Different Commercial Serological Tests for the Detection of Toxoplasma gondii Antibodies in Serum of Naturally Exposed Pigs

Toxoplasma gondii is the aetiological agent of the zoonotic disease toxoplasmosis and transmitted among other ways by chemically and physically untreated, that is, raw pork to humans. The detection of Toxoplasma gondii is impossible by currently practiced meat inspection, but serological tests can be used to detect Toxoplasma gondii antibodies in pig herds and can consequently be helpful to identify potentially contaminated pork. Therefore, appropriate serological tests are required. In this study, serum samples of 1368 naturally exposed slaughter pigs from 73 Austrian farms were collected. Serum samples of at least 16 slaughter pigs per farm were tested. The prevalence of Toxoplasma gondii antibodies in serum was measured by a commercial available modified agglutination test (MAT) and compared to three different commercial available enzyme‐linked immunosorbent assays (ELISA). The MAT detected 6.5%, ELISA I 6.7%, ELISA II 4.8% and ELISA III 4.3% of the pigs as Toxoplasma gondii antibody positive. The agreement, according to the kappa coefficient (κ), was substantial between the MAT and ELISA I (κ = 0.62), II (κ = 0.64) and III (κ = 0.67). A better agreement was determined between ELISA I and II (κ = 0.715), ELISA I and III (κ = 0.747) and ELISA II and III (κ = 0.865). At least one pig per farm was detected Toxoplasma gondii antibody positive in 17 (23.3%) farms by the MAT, 26 (35.6%) farms by ELISA I, 16 (21.9%) farms by ELISA II and 11 (15.1%) farms by ELISA III. Pig farms with a high number of Toxoplasma gondii antibody‐positive pigs or high antibody titres were identified by all of the four used serological tests. Concerning the occurrence of Toxoplasma gondii antibodies in Austrian pig farms, a monitoring and surveillance programme would be reasonable to find high‐risk farms.     

Toxoplasma gondii infection in domestic ducks, free-range and caged chickens in southern China

Toxoplasma gondii is widely distributed in humans and other animals including domestic poultry throughout the world, but little is known of the prevalence of T. gondii in chickens and ducks in People's Republic of China. In the present study, antibodies to T. gondii were investigated in 349 domestic ducks (Anas spp.), 361 free-range, and 244 caged chickens (Gallus domesticus) raised in commercial flocks in Southern China's Guangdong Province using the modified agglutination test (MAT). Antibodies to T. gondii (MAT titer of 1:5 or higher) were found in 56 (16%) of 349 ducks, 41 (11.4%) of 361 free-range, and 10 (4.1%) of 244 caged chickens. The results indicate soil contamination due to T. gondii oocysts because free-range chickens feed from the ground, and suggest that the meat from the domestic poultry may be an important source for human infection by T. gondii in People's Republic of China.     

Seroprevalence of Toxoplasma gondii infection in sheep and alpacas (Llama pacos) in Chile

Serum samples from 408 sheep from different regions of Chile and 447 alpacas (Llama pacos) from the north of the country were tested for Toxoplasma gondii antibodies. The indirect haemagglutination test (IHAT) was used in both species and the indirect immunofluorescence test (IIFT) was also used on the sheep samples in order to compare the performance of the tests in that species. In both tests, titers ≥1:16 were considered diagnostically significant. Sera from 49 sheep (12%) were positive to T. gondii antibodies by the IHAT. When using the IIFT, 114 sheep sera (28%) were positive. The different results obtained in sheep sera between the tests were significant (p<0.0001). No differences were observed between geographical locations or sex of the sampled sheep regarding serological detection of T. gondii antibodies in sheep. As expected, adult sheep showed higher T. gondii reactivity than young sheep (p=0.0008). The corrected prevalence of toxoplasmosis in alpaca was 16.3% (32 positive out of 447). The rather low prevalence in alpacas may be associated with their extensive management as well as the extreme climatic conditions of The Andes which apparently would not be favorable for the transmission of the parasite.     

Prevalence of <Emphasis Type="Italic">Toxoplasma gondii</Emphasis> and <Emphasis Type="Italic">Neospora caninum</Emphasis> infections in sheep from Federal District,central region of Brazil

Serum samples from 1028 sheep were collected from 32 herds within Federal District, in the central region of Brazil. The samples were examined by indirect fluorescent antibody test (IFAT) using sera diluted 1:64 and 1:50 as cut-off values for the detection of antibodies against Toxoplasma gondii and Neospora caninum, respectively. The observed prevalence for T. gondii infection was 38.22% (26.81%<CI 0.95<49.62%), and the titers ranged from 64 to 65536. The observed prevalence for N. caninum infection was 8.81% (7.08%<CI 0.95<10.53%). The titers ranged from 50 to 51200. The reactant sera to both pathogens corresponded to 4.67% of the samples. The risk factors were not determined because of the absence of negative herds for T. gondii and the high proportion of positive herds for N. caninum (87.50%). The prevalence for T. gondii infection was significantly higher among males than in females. The present work is the first report on seroprevalence of T. gondii and N. caninum in sheep from Federal District and shows that infection by both parasites is widespread in the ovine population from this region.     

Activities of adenine nucleotide and nucleoside degradation enzymes in platelets of rats infected by Trypanosoma evansi

Pigeons (Columba livia) cohabit with humans in urban and rural areas, representing a public health problem since microorganisms are transmitted through the inhalation of dust from their dry feces (chlamydiosis) and through ingestion of their undercooked or poorly refrigerated meat (toxoplasmosis). This study aimed to evaluate the presence of Chlamydophila psittaci and Toxoplasma gondii in pigeons from four cities in São Paulo State, Brazil. C. psittaci was evaluated through hemi-nested polymerase chain reaction (hnPCR) using cloacal and tracheal swabs, whereas T. gondii specific antibodies were assessed by means of modified agglutination test (MAT), mouse brain and muscle bioassay, and polymerase chain reaction (PCR). To confirm the infection in mice, T. gondii antibodies were assayed by using indirect fluorescent antibody test (IFAT). Considering C. psittaci, 40/238 (16.8%; 95%CI 12.6–22.1%) samples were positive according to hnPCR, especially for the cities of São Paulo (42.5%) and Bauru (35%). As regards T. gondii, 12/238 (5%; 95%CI 2.9–8.6%) serum samples were positive according to MAT. Of these, five samples had titer equal to 1:8; six samples, 1:16; and one sample, 1:32. Bioassay, IFAT and PCR were negative for mouse toxoplasmosis. The absence of T. gondii antibodies suggests that pigeons may be infected with a low concentration of the agent, not detected by the antigen test. Thus, C. psittaci represents an actual problem concerning bird health.     

Toxoplasma gondii in Ireland: Seroprevalence and Novel Molecular Detection Method in Sheep,Pigs, Deer and Chickens

Toxoplasma gondii is among the most studied parasites worldwide but there is not much information about it published in Ireland. The objectives of this study were to determine the seroprevalence of T. gondii in sheep, pigs, deer and chickens and the molecular detection of T. gondii DNA in muscle tissue. Serum samples were collected from these species at the time of slaughter at Irish abattoirs during 2007 and tested for anti‐T. gondii antibodies using a commercial semi‐quantitative latex agglutination test. Antibodies (titre ≥1 : 64) were found in 36% (105/292) sheep, 4.7% (15/317) pigs and 6.6% (23/348) deer. In chickens, 18% (65/364) had antibody titres, ranging between 1 : 5 and 1 : 1024. Significant (P ≤ 0.05) age‐related differences in seroprevalence were found in adult sheep (58.1%) and pigs (23.1%). Significant gender differences in seroprevalence was also found in sheep with more females (43%) than males (22.4%) being positive. However, when adjusted for age through logistic regression gender was no longer significant. Seroprevalence was also evaluated on farm locations grouped to NUTS level 3, but the prevalence was too low to draw any statistical conclusions. Using a nested PCR, the presence of T. gondii DNA was detected in diaphragm samples from 3.6% (3/83) sheep, 13.0% (3/23) pig and 4.2% (3/71) deer. Meat digestion liquids from a Trichinella spp. survey in pigs were also used for the first time to detect T. gondii. Toxoplasma gondii DNA was detected in 50% (10/20) of pooled samples. This is the first in depth study of T. gondii seroprevalence in animals in Ireland and a novel method, using digestion liquid from pooled diaphragm samples, for PCR detection in pigs is described.     

Leukocyte counts in bronchoalveolar lavage fluid obtained from normal and Maedi–Visna‐infected sheep

Background: Bronchoalveolar lavage has proven helpful for the diagnosis of certain ovine diseases of the lungs. There is insufficient data concerning the leukocyte profile of bronchoalveolar lavage fluid (BALF) from MaediVisna infected sheep. Objective: The objective of this study was to assess the differential leukocyte profile of BALF associated with Maedi virus infection in sheep and to determine whether cytologic examination of BALF is an effective way to diagnose Maedi or determine the severity of lung lesions. Methods: BALF and serum samples were analyzed from 400 sheep. Sediment smears of bronchoalveolar lavage were stained with Diff‐Quik and examined microscopically to obtain a 200‐cell differential cell count. Serum was tested using a commercial kit for Maedi–Visna virus antibodies. Lung samples obtained at the time of slaughter were weighed and examined histologically. Results: Maedi‐infected sheep (n=267; seropositive with lung lesions) had a significantly higher percentage of lymphocytes and lower percentage of macrophages in BALF than normal sheep (n=133; seronegative and no lung lesions). These differences were significantly more severe in animals with advanced vs moderate lung lesions. Using classification trees, a cut‐off of 13.5% lymphocytes was predictive of Maedi infection and a cut‐off of 24.5% lymphocytes was predictive of advanced lung lesions. Conclusions: Cytologic examination of BALF is useful for the clinical diagnosis of Maedi in sheep and provides important information about the severity of the lung lesions.     

Prevalence of Toxoplasma infection in sheep in Romania

Toxoplasma antibodies have been demonstrated in 37.08 and 31.07% among 502 sheep by means of indirect immunofluorescent antibody and indirect haemagglutination tests, respectively, from three localities of Romania. Ninety-six serum samples out of 100 found positive by indirect immunofluorescence also reacted positive when subjected to Sabin-Feldman dye test. Toxoplasma gondii was isolated from two of 13 serologically positive sheep and also from the organs of a sheep collected from the slaughter-house. It is suggested that ovine toxoplasmosis may be a substantial public health problem in Romania.     

An outbreak of Toxoplasmosis amongst squirrel monkeys in an Israeli monkey colony

An outbreak of Toxoplasmosis in a colony of squirrel monkeys (Saimiri sciureus) in Israel is described. Serological, pathological, and molecular findings of monkeys, as well as rodents and pigeons from the vicinity are summarized. Seventy-nine percent (19/24) of monkeys were T. gondii seropositive at titer 1:16 whilst 4% (1/24) were also seropositive at titer 1:64 using the Modified Agglutination Test (MAT). Eighty four percent (21/25) of rats were positive at titer 1:16 and 8% (2/25) of rats were positive at titer 1:32. DNA amplification of a 529 bp repeated sequence of T. gondii was detected in the liver and lungs of all monkeys tested, 6/7 in myocardial extractions and 5/6 in brain extractions.     

First identification of Sarcocystis tenella (Railliet, 1886) Moulé, 1886 (Protozoa: Apicomplexa) by PCR in naturally infected sheep from Brazil

Sarcocystis tenella is a dog–sheep protozoan parasite, causing a widespread enzootic muscle parasitosis and neurological disease mainly in lambs. This parasite is pathogenic to sheep and important to the economical production of sheep. The present study was initially aimed to determine Toxoplasma gondii infection and the occurrence of co-infection with other Apicomplexa parasites in 602 Brazilian sheep. Twenty of these sheep were positive with antibodies to T. gondii by MAT and IFAT-IgG tests, positive with PCR-RFLP genotyping at multiple loci, and parasites were isolated from mice infected with sheep tissue samples. Two additional sheep born in Brazil, a 2-year-old female Polwarth (Ideal) sheep, a breed originated from Australia (#1), and a 1-year-old male Corriedale sheep, a breed originated from New Zealand and Australia (#2) were positive to T. gondii antibodies by serum tests, and PCR, but negative for bioassay in mice. In genotyping at 12 loci, sheep #1 sample and #2 presented positive results only for some markers. PCR-RFLP of 18S ribosomal RNA (18S rRNA) was performed in all 22 animals to identify the possibility of co-infection of T. gondii with other Apicomplexa parasites, such as S. tenella, Neospora caninum and Hammondia hammondi, resulting in a T. gondii profile for the first 20 animals and a unique genotyping profile for sheep #1 and #2, identical to S. tenella. The 18S rRNA PCR products (310 bp) were sequenced and blasted to GenBank database at NCBI. Both samples were identical to S. tenella 18S rRNA gene (GenBank accession number L24383-1). These results suggest the existence of co-infection of S. tenella with T. gondii in ewes from Brazil.     

Seroprevalence and risk factors of caprine toxoplasmosis in Minas Gerais, Brazil

The objective of this work was to carry out a study on caprine toxoplasmosis in the state of Minas Gerais, Brazil. To determine the prevalence of toxoplasmosis in goats in Minas Gerais, 767 sera from goats were tested by ELISA (enzyme-linked immunosorbent assay) and IFAT (indirect fluorescence antibody test). The prevalence of antibodies to Toxoplasma gondii was 43.0% and 46.0% by ELISA and IFAT, respectively. It was observed that 26.8% of the goats show low-avidity IgG to T. gondii. These results suggest the presence of animals in recent phase of toxoplasmosis in Minas Gerais. The risk factors for toxoplasmosis in goats were: age over 36 months (OR = 1.21; IC 95% 1.02–1.44), use of pen (OR = 1.83; IC 95%1.01–3.31) and pure breed animals (OR = 2.49; IC 95% 1.11–5.59).     

Seroprevalence of Toxoplasma gondii infection in domestic sheep and goats in Borno state,Nigeria

Serum samples were collected from 372 sheep and same number of goats from the three geopolitical zones of Borno state, Nigeria. The samples were tested for the presences of Toxoplasma gondii antibodies by enzyme-linked immunosorbent assay. Of these, 6.7% (25/372) and 4.6% (17/372) of sheep and goats, respectively, were found to be seropositive to T. gondii antibodies, both far less than the estimated global average of 31%. Results were statistically analyzed by chi-square (χ²) test. The results showed that age, environmental conditions, and farm location are the main determinants of prevalence of antibodies against T. gondii in the study area. Older animals (>3 years) are significantly more infected than younger animals (between 6 months and 1 year).The prevalence of anti T. gondii antibodies is significantly higher (P < 0.05) in both sheep and goats sampled from the southern zone than the northern zone. Animals from the southern zones are about four times more likely to be exposed to T. gondii infection than those in the northern zone, (sheep; odds ratio (OR) = 4.25, 95% confidence interval (CI) = 1.177–15.36, P = 0.018), (goats; OR = 4.38, 95% CI = 0.925–20.73, P = 0.04). Farm location in urban area was identified as a risk factor for sheep (OR = 6.06, 95% CI = 2.53–14.54, P = 0.000), and goats (OR = 4.99, 95% CI = 1.59–15.62, P = 0.004). Current data on prevalence of ovine and caprine T. gondii in Borno state are provided by the study as well as identifying the main risk factors associated with T. gondii infection in the area.     

Toxoplasma gondii Infections in Chickens (Gallus domesticus): Prevalence,Clinical Disease,Diagnosis and Public Health Significance

Chickens are considered one of the most important hosts in the epidemiology of Toxoplasma gondii infection because they are an efficient source of infection for cats that excrete the environmentally resistant oocysts and because humans may become infected with this parasite after eating undercooked infected chicken meat. The objective of this study is to review worldwide prevalence of T. gondii infection in chickens and to assess the role of infected chickens in the epidemiology of toxoplasmosis in humans. A very high prevalence of the parasite was found in chickens raised in backyards (up to 100%) and free‐range organic (30–50%) establishments.     

Immunohistochemical identification of Toxoplasma gondii in tissues from Modified Agglutination Test positive sheep

Toxoplasma gondii is a zoonotic agent of great importance in veterinary and public health. The aim of this study was to identify T. gondii by IHC (immunohistochemistry) in different sheep tissues and to determine if an association exists between the results obtained by this method and those obtained by the Modified Agglutination Test (MAT). Tissue specimens of twenty-six sheep seroreactive for T. gondii were selected for histopathological evaluation. The presence of T. gondii was investigated in brain, liver and heart samples by IHC and a possible anti-T. gondii antibody cross reactions with other parasites. McNemar's, Chi-square and Fisher's Exact Tests were applied for the statistical analysis of the results. The analysed tissues showed at least one of the following histopathological changes: mild-to-moderate congestion, focal polymorphonuclear inflammatory infiltrate and multifocal or focal mononuclear inflammatory infiltrate. Sarcocystis spp. were identified in the histological sections from both the heart and diaphragm tissues of 88.5% (23/26) of the animals. A total of 46.2% (12/26) of the T. gondii seroreactive sheep was also positive for T. gondii by IHC in at least one organ (brain, liver or heart). The liver IHC-positivity for T. gondii was statistically equivalent to the global individual IHC-positivity, according to McNemar's test. In addition, IHC allowed the detection of T. gondii in infected animals regardless of the titration observed in the MAT. The statistical difference observed between the three organs when comparing the low titration group, suggested that the heart might be the most suitable organ to detect T. gondii infection by IHC. The IHC results in this study revealed that almost half of MAT positive animals could serve as potential sources of infection for humans because bradyzoites were identified in different tissues, regardless of the MAT titration.     

Shedding and Seroprevalence of Pathogenic Leptospira spp. in Sheep and Cattle at a New Zealand Abattoir

A cross‐sectional study was carried out on sheep and cattle slaughtered at a New Zealand abattoir from September to November 2010 to investigate the supplier‐specific shedding rate, renal carriage rate and seroprevalence of leptospires. In the 2008/2009 season, this abattoir experienced three human leptospirosis cases from 20 staff, of which two were hospitalized. Urine, kidney and blood samples were collected from carcasses of 399 sheep (six suppliers, 17 slaughter lines) and 146 cattle (three suppliers, 22 slaughter lines). The urine and kidney samples were tested by quantitative real‐time PCR (qPCR), while serum samples (from coagulated blood samples) were tested by microscopic agglutination test (MAT). In total, 27% (73/274; 95% CI: 18–37) of urine samples tested positive by qPCR. Species‐specific shedding rates (prevalence of positive urine qPCR) were 31% (95% CI: 17–48) for sheep and 21% (95% CI: 14–30) for cattle. For 545 kidney samples tested, 145 were qPCR positive (27%; 95% CI: 17–39). The average prevalence of kidney qPCR positivity was 29% (95% CI: 17–45) for sheep and 21% (95% CI: 15–28) for cattle. Three hundred and thirty of 542 sampled sheep and cattle had antibodies against Leptospira borgpetersenii serovar Hardjobovis (Hardjobovis) and/or Leptospira interrogans serovar Pomona (Pomona), based on reciprocal MAT titre ≥1 : 48 (overall seroprevalence of 61%; 95% CI: 48–73). Seroprevalence was 57% (95% CI: 40–72) for sheep and 73% (95% CI: 59–83) for cattle. Among the seropositive animals, 41% (70/170; 95% CI: 30–54) were shedding (tested positive by urine qPCR) and 42% (137/330; 95% CI: 30–54) had renal carriage (tested positive by kidney qPCR). Some risk management options for abattoirs or farms to prevent human leptospirosis infections include vaccination of maintenance hosts, the use of personal protective equipment, and the application of urine qPCR to detect shedding status of stock as surveillance and as an alert.     

The epidemiological interpretation of serological responses to leptospiral serovars in sheep.

Serum samples were examined for evidence of leptospiral agglutinins from 928 sheep from 45 lines and kidneys from 12 of these lines for evidence of leptospiral infection. All sheep had been submitted for slaughter at meat works in the Manawatu.

Serological results were analysed using the results at a minimum serum dilution in the microscopic agglutination test (MAT) of 1:24 and at a minimum dilution of 1:48. It was shown that a minimum dilution of 1:24 resulted in many non-specific or cross-reactions. A minimum dilution of 1:48 was more accurate for detecting the serological prevalence of specific agglutinins to leptospires in ovine sera. Twenty percent of the sheep had titres of 1:48 or greater to hardjo, 3.8% to pomona, 2.6% to tarassovi, 2.3% to copenhageni and 2.7% to ballum. No titres of 1:48 or greater to australis were detected. Serovar hardjo was isolated from the kidneys of three animals in one line.

Eighteen months later 291 serum samples and 95 urine samples were collected from live animals on the property from which the three hardjo infected animals originated. No titres to hardjo were detected in the sera of lambs, but a serological prevalence of 44% and 84% to this serovar was demonstrated in the hoggets and ewes respectively. No leptospires were demonstrated in any of the urine samples.

These results show that sporadic infection of sheep with hardjo can occur but they also indicate that infection with this serovar is not endemic and that sheep are unlikely to act as maintenance hosts for hardjo in New Zealand.