A novel low-cost method for Mycobacterium avium subsp. paratuberculosis DNA extraction from an automated broth culture system for real-time PCR analysis

PCR is a highly accurate technique for confirming the presence of Mycobacterium avium subsp. paratuberculosis (Map) in broth culture. In this study, a simple, efficient, and low-cost method of harvesting DNA from Map cultured in liquid medium was developed. The proposed protocol (Universidad Austral de Chile [UACH]) was evaluated by comparing its performance to that of two traditional techniques (a QIAamp DNA Stool Mini Kit and cethyltrimethylammonium bromide [CTAB] method). The results were statistically assessed by agreement analysis for which differences in the number of cycles to positive (CP) were compared by Student''s t-test for paired samples and regression analysis. Twelve out of 104 fecal pools cultured were positive. The final PCR results for 11 samples analyzed with the QIAamp and UACH methods or ones examined with the QIAamp and CTAB methods were in agreement. Complete (100%) agreement was observed between data from the CTAB and UACH methods. CP values for the UACH and CTAB techniques were not significantly different, while the UACH method yielded significantly lower CP values compared to the QIAamp kit. The proposed extraction method combines reliability and efficiency with simplicity and lower cost.     

PCR-based detection of Toxoplasma gondii from cattle in southern Iran

ObjectiveToxoplasma gondii is a protozoan parasite that is widely prevalent in most warm-blooded vertebrates. Humans mainly become infected by eating raw or undercooked meat. This study was designed to investigate the infection of cattle with T. gondii in Jahrom, southern Iran.MethodsTissue samples consisting of heart, diaphragm, and tongue were collected from 125 slaughtered cattle. DNA samples were extracted from the homogenized tissues. T. gondii was detected and genotyped using nested-polymerase chain reaction (Nested-PCR) and polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) based on GRA6 and SAG2 (3', 5' terminal regions) genes, respectively.ResultsThe prevalence of T. gondii DNA was 56% in cattle. The most infected tissue was the diaphragm (54.4%) followed by the heart (48.8%) and tongue (43.2%). Type II was the most prevalent genotype (70%) among T. gondii isolates.ConclusionIn this study, the high prevalence of T. gondii infection in cattle meat indicates the important role of cattle in the transmission of infection to humans. Therefore, incorporating the correct method of consuming meat in health education programs is crucial to prevent human infection.     

Determination of an efficient and reliable method for DNA extraction from ticks

Molecular detection of pathogenic microorganisms in ticks is based on DNA amplification of the target pathogen; therefore, extraction of DNA from the tick is a major step. In this study, we compared three different tick DNA extraction protocols based on an enzymatic digestion by proteinase K followed by DNA extraction by a commercial kit (method 1), or on mortar crushing, proteinase K digestion and phenol/chloroform DNA extraction (method 2) and fine crushing with a beads beater, proteinase K digestion and DNA extraction using a commercial kit (method 3). The absence of PCR inhibitors and the DNA quality were evaluated by PCR amplification of the tick mitochondrial 16S rRNA gene using tick-specific primers. With method 1, 23/30 (77%) of the samples were extracted; with method 2, 30/31 (97%) of the samples were extracted and with method 3, 30/30 (100%) of the samples were extracted. DNA extraction efficiency using method 3 is significantly higher than DNA extraction efficiency using method 1 (100% versus 77%, P < 0.05). There was no significant difference between methods 2 and 3. Method 3 was however more adapted to cohort studies than method 2. This technique was validated for cohort tick DNA extraction and applicable to the treatment of small samples such as nymphs and soft ticks with 100% efficiency.     

Evidence for widespread Leishmania infantum infection among wild carnivores in L. infantum periendemic northern Spain

Leishmania spp. infection was investigated in tissue samples of wild carnivores from the Spanish Basque Country (BC), by PCR and DNA sequencing. The region is at the northern periphery of Leishmania infantum endemic Iberian Peninsula and infection in the dog (reservoir) or other species has not been previously reported. Leishmania kinetoplast DNA was detected by real-time PCR (rtPCR) in 28% (44/156) of animals. Specifically, in 26% of Eurasian badgers (n = 53), 29% of foxes (n = 48), 29% of stone martens (n = 21) and in 25–50% of less numerous species including genets, wild cats, pole cats, European mink and weasels. Infected animals particularly badgers, were most prevalent in the southernmost province of the BC (Araba) in areas dominated by arable land. Subsequent amplification and sequencing of a fragment of the rRNA internal transcribed spacer 2 (ITS2) from a subset of rtPCR positives samples confirmed the species as L. infantum, showing a high sequence homogeneity with ITS2 sequences of L. infantum from dogs and humans from southern Spain. In summary, this study reports for the first time L. infantum infection in wild carnivores from the BC including in stone martens, pole cats and minks in which infection has not been previously described. It supports the need to study infection in dogs and people in this region and is an example of the value of infection surveillance in wildlife to assess potential risks in the domestic environment and their role in spreading infections in non-endemic areas.     

Evaluation of PCR inhibitory effect of enrichment broths and comparison of DNA extraction methods for detection of Salmonella Enteritidis using real-time PCR assay

The best enrichment broth and DNA extraction scheme was determined for rapid and sensitive detection of Salmonella Enteritidis in steamed pork using real-time PCR. The inhibitory effect of commonly used Salmonella enrichment broths, Rappaport-Vassiliadis (RV) and Muller-Kauffmann tetrathionate with novobiocin (MKTTn), on real-time PCR was confirmed. The inhibition of PCR was statistically significant (p < 0.05) in RV and MKTTn, as compared with buffered peptone water (BPW) or phosphate-buffered saline. The inhibitory effect of the selective enrichment media was successfully removed by using a modified DNA extraction, PrepMan Ultra Reagent with an additional washing step or the DNeasy Tissue Kit. In three experiments, when applied to detection of Salmonella Enteritidis in steamed pork, the real-time PCR coupled with single 24 h enrichment with BPW performed better than double 48 h enrichment with BPW plus RV or MKTTn. The simple real-time PCR assay using BPW proved to be a rapid and sensitive test for detection of low concentrations of Salmonella Enteritidis in steamed pork samples as compared with the conventional culture method.     

Detection of Toxoplasma gondii in Crassostrea spp. oysters cultured in an estuarine region in eastern Amazon

The aim of the present study was to detect DNA of Toxoplasma gondii in Crassostrea spp. oysters cultured in the state of Pará, Brazil. A total of 400 oysters were directly collected from a fixed rack system. Gills, gastrointestinal tract (GIT) and intervalvular liquid were separated and grouped into pool samples of 10 animals, resulting in 40 samples each of gills, GIT and intervalvular liquid. DNA extraction was performed using a commercial kit, and T. gondii DNA was detected by nested PCR using the primers Toxo3 and Toxo4, which produced an amplification product of 155 bp of the T. gondii gene B1. Nucleotide sequencing was performed for positive samples, and the obtained sequences were identified by comparison with sequences in GenBank. The DNA of T. gondii was detected in 5.8% (7/120) of the pool samples, of which 7.5% (3/40) was in the GIT, 5% (2/40) in the gills, and 5% (2/40) was in the intervalvular liquid. The obtained sequences presented 100% identity and overlap with T. gondii DNA sequences. This is the report of detection of T. gondii DNA in oysters from genus Crassostrea spp. originating from the state of Pará, eastern Amazon.     

Cytologic findings, and feline herpesvirus DNA and Chlamydophila felis antigen detection rates in normal cats and cats with conjunctival and corneal lesions

Samples were collected from 36 cats with feline herpesvirus (FHV-1)-related ocular disease (conjunctivitis, epithelial or stromal keratitis, or corneal sequestration), and 17 cats without ocular changes. Corneoconjunctival swabs, scrapings and biopsies were tested in various combinations for presence of FHV-1 DNA using single round (sr) polymerase chain reaction (PCR) and nested PCR (nPCR). Additional swabs from the inferior conjunctival fornix were tested by enzyme-linked immunosorbent assay for Chlamydophila felis antigen. Cytologic evaluation was carried out on conjunctival (cats with conjunctivitis) and corneal (cats with keratitis) cytobrush preparations. FHV-1 DNA was detected by PCR in 14 (39%) cats with ocular disease and 1 (6%) of the control group. Agreement between srPCR and nPCR results was significant (P < 0.01). FHV-1 DNA was detected in 3/7 cats with conjunctivitis, 5/6 cats with epithelial keratitis, 3/11 cats with stromal keratitis, and 3/12 cats with corneal sequestration. There was a significant association (P = 0.0027) between viral presence and epithelial keratitis. However, no significant association was found between viral presence and conjunctivitis (P = 0.059), stromal keratitis (P = 0.15), or corneal sequestration (P = 0.18). With respect to FHV-1 DNA detection, intersample agreement was significant (P < 0.03). No sampling technique seemed more likely than another to harvest detectable viral DNA, except for cats with corneal sequestrum in which viral DNA was not detected using corneoconjunctival swabs. FHV-1 DNA was detected in 6/9 samples with intranuclear inclusion bodies and in 6/7 cats with eosinophils on cytologic examination. All samples tested negative for C. felis antigen.     

Factors affecting seroprevalence of Toxoplasma gondii in the endangered Iberian lynx (Lynx pardinus)

Wild felids are considered important in maintaining the sylvatic cycle of Toxoplasma gondii. Although, T. gondii antibodies have been reported in several species of wild felids, little is known of the epidemiology and risk factors associated with T. gondii infection in wild cats. The Iberian lynx (Lynx pardinus) is the most endangered felid species in the world. In the present study, seroprevalence and associated risk factors for T. gondii infection in a large population of Iberian lynx in Spain were determined. Serum samples from 129 Iberian lynx collected from 2005 to 2009 and 85 wild rabbits (Oryctolagus cuniculus), sharing the habitat with the Iberian lynx, were tested for antibodies to T. gondii by the modified agglutination test (MAT) using a cut-off value of 1:25. Antibodies to T. gondii were found in 81 of 129 (62.8%) Iberian lynx. Seroprevalence to T. gondii in Iberian lynx significantly increased with age (P < 0.001). T. gondii seroprevalences were similar in free-ranging (66.7% of 93) and wild-caught captive lynx (69% of 84) but significantly lower in captive-born lynx (22.5% of 40). Seroprevalence was higher in lynx with concurrent Cytauxzoonfelis (88% of 25) but not with concurrent Feline Leukemia Virus (FeLV) infection (53.8% of 13). There were no significant differences in seroprevalence between sexes, geographic region and year of sample collection (2005–2009). Oocysts of T. gondii were not detected microscopically in fecal samples from 58 lynx. Wild rabbits are considered the most important food for the lynx. Antibodies to T. gondii were found in 14 (11.9%) of 85 rabbits tested. The present results indicate that T. gondii infection is widespread in the two areas where Iberian lynx survive in Spain. The fact that four captive-born lynx seroconverted was indication of contact with T. gondii also in the Captive Breeding Centers, hence, control measures to prevent T. gondii infection would be necessary in these centers.     

山羊绒毛干中线粒体DNA和核DNA的提取与PCR扩增

本研究以PCR缓冲液、MgCl2溶液和蛋白酶K为裂解提取液,建立了一种从山羊绒毛干中快速提取DNA的方法。结果表明,该方法不仅能从山羊绒毛干中提取出线粒体DNA和核DNA,而且能够有效地进行线粒体基因和核基因组基因的PCR扩增,为拓展羊绒样品在个体识别、遗传资源评价、分子进化和分子标记辅助育种等研究领域中的应用奠定基础。     

Feline Herpesvirus 1 and Mycoplasma spp. Conventional PCR Assay Results From Conjunctival Samples From Cats in Shelters With Suspected Acute Ocular Infections

Signs of ocular infections like discharge and conjunctivitis occur commonly in cats in shelters and feline herpesvirus 1 (FHV-1), Chlamydia felis, Mycoplasma spp, and feline calicivirus (FCV) are thought to be the most common causes. While molecular assays are available to amplify nucleic acids of each of these agents as single tests or in panels, additional information is needed concerning whether the assay results can be used to predict response to treatment. The objectives of this study were to report results for conventional polymerase chain reaction (PCR) assays that amplify nucleic acids of FHV-1, Mycoplasma spp., C. felis, and FCV from cats with signs of acute ocular and upper respiratory infections in an animal shelter and to determine whether the results are associated with treatment responses to topical administration of cidofovir (anti-FHV-1) or oxytetracycline (anti-Mycoplasma spp. and C. felis). Conjunctival samples were collected from both eyes of 60 cats with ocular signs of disease. Total deoxyribonucleic acid (DNA) and ribonucleic acid (RNA) were extracted from each sample and assayed for DNA of FHV-1, Mycoplasma spp., and C. felis and RNA of FCV by conventional PCR assays. Cats were randomized to be administered either oxytetracycline ointment or cidofovir drops in both eyes and a standardized ocular disease score system was used to determine a total ocular score for each cat prior to treatment on Day 0 and on Day 7. Nucleic acids of one or more agents were amplified from one or both eyes from 39 of 60 cats (65%). FHV-1 DNA (21 cats), Mycoplasma spp. DNA (25 cats) or FCV RNA (2 cats) were amplified most commonly. After treatment for 7 days, 32 of 60 cats (53.3%) were considered improved with 27 of 32 cats (84.4%) having ocular scores of 0 (21 cats) or 1 (6 cats). When the results of the FHV-1 PCR assay were compared to cidofovir treatment responses, the positive and negative predictive values of the assay were shown to be 29.4% and 60%, respectively. When the results of the Mycoplasma spp. PCR assay were compared to oxytetracycline treatment responses, the positive and negative predictive values of the assay were shown to be 40% and 38.5%, respectively. The predictive value of conventional PCR assay results for FHV-1 or Mycoplasma spp. DNA was low, suggesting that performing these tests to formulate a treatment protocol has minimal clinical utility in cats with suspected acute ocular infections.     

Optimized protocol for multiplex nested polymerase chain reaction to detect and differentiate Haemophilus parasuis,Streptococcus suis,and Mycoplasma hyorhinis in formalin-fixed,paraffin-embedded tissues from pigs with polyserositis

An optimized protocol was developed for the simultaneous detection and differentiation of Haemophilus parasuis, Streptococcus suis, and Mycoplasma hyorhinis in formalin-fixed, paraffin-embedded (FFPE) tissues with multiplex nested polymerase chain reaction (PCR). This method also determines the prevalence of these bacteria in pigs with polyserositis. DNA extraction with a combination of a commercial reagent and proteinase K resulted in more frequent detection of the pathogens than DNA extraction with proteinase K alone. Among FFPE tissue samples from 312 cases of polyserositis in which at least 1 bacterial species was detected, multiplex nested PCR detected H. parasuis in 239 (77%), S. suis in 124 (40%), and M. hyorhinis in 40 (13%). The disease was caused by a single pathogen in 224 (72%) of the cases and multiple pathogens in 88 (28%). Among the pigs positive for H. parasuis, S. suis, and M. hyorhinis by multiplex nested PCR, the pathogen was isolated from only 11%, 35%, and 28%, respectively. Therefore, the PCR protocol developed in this study is a useful diagnostic method when samples are negative after isolation methods and even for samples in which only 1 pathogen was isolated.     

Seropositivity of Toxoplasma gondii in domestic donkeys (Equus asinus) and isolation of T. gondii from farm cats

Donkeys (Equus asinus) are used as both companion and working animals throughout the world and in some countries, their meat and milk are used for human consumption. Here we report the first serological survey of Toxoplasma gondii in donkeys in the United States. Serum samples from 373 donkeys from eight farms in five states were tested for T. gondii antibodies by the modified agglutination test (MAT). Twenty-four of 373 (6.4%) of donkeys were seropositive, with MAT titers ranging from 25 to ≥200. All seropositive donkeys were Miniature breed. Seropositivity prevalence was 7.0% in female donkeys (20/282) and 4.1% in male donkeys (4/91). No donkeys less than 24 months of age (129) were seropositive, suggesting postnatal transmission of infection. Domestic cats were present on six of the eight farms. Three cats from one farm had MAT titers of 200. Viable T. gondii was isolated from the hearts of two cats, but not from brain tissues. Genotyping of isolate DNA extracted from culture-derived tachyzoites using 10 PCR-restriction fragment length polymorphism (RFLP) markers (SAG1, SAG2, SAG3, BTUB, GRA6, c22-8, c29-2, PK1, L358 and Apico loci) revealed that both isolates were clonal Type II (ToxoDB PCR-RFLP genotype #1). This is the first serological survey for T. gondii in donkeys in the United States, and suggests that donkey milk and meat should be considered as a potential source for human infection. The role of barn cats in the transmission of T. gondii to donkeys on farms warrents further investigation.     

Feline immunodeficiency virus, feline leukemia virus and Toxoplasma gondii in stray and household cats in Kerman-Iran: Seroprevalence and correlation with clinical and laboratory findings

This study was carried out to determine the seroprevalence of feline leukemia virus (FeLV), feline immunodeficiency virus (FIV) and Toxoplasma gondii (T. gondii) infection among stray and owned cats in southeastern Iran and to identify the influence of age, sex, lifestyle, health status, and laboratory findings on seropositivity. The overall infection rate for FIV, FeLV, and T. gondii was 19.2%, 14.2%, and 32.1% respectively. Results of the multivariate logistic regression analysis showed that old adults more likely to be seropositive than juveniles for FIV, FeLV, and T. gondii (adjusted odds ratios [ORs], 1.84, 1.56, and 2.57 respectively). Anemic and diseased cats ([ORs], 6.62 and 0.9) were at a greater risk of testing positive for FeLV. Male cats were 4.91 times as likely to have FIV as were female and hyperglobulinemia was significantly more prevalent in FIV-infected cats ([ORs], 3.4). In conclusion, FIV and FeLV seem to be endemic in Iran and retroviral-associated immunosuppression may be a risk factor for active toxoplasmosis in infected cats.     

Direct Detection of Mycobacterium bovis in Bovine Lymph Nodes by PCR

Thirty‐five lymph node samples were taken from animals with macroscopic lesions consistent with Mycobacterium bovis infection. The animals were identified by postmortem examination in an abattoir in the northwestern region of state of Paraná, Brazil. Twenty‐two of the animals had previously been found to be tuberculin skin test positive. Tissue samples were decontaminated by Petroff’s method and processed for acid‐fast bacilli staining, culture in Stonebrink and Lowenstein‐Jensen media and DNA extraction. Lymph node DNA samples were amplified by PCR in the absence and presence (inhibitor controls) of DNA extracted from M. bovis culture. Mycobacterium bovis was identified in 14 (42.4%) lymph node samples by both PCR and by culture. The frequency of PCR‐positive results (54.5%) was similar to that of culture‐positive results (51.5%, P > 0.05). The percentage of PCR‐positive lymph nodes increased from 39.4% (13/33) to 54.5% (18/33) when samples that were initially PCR‐negative were reanalysed using 2.5 μl DNA (two samples) and 1 : 2 diluted DNA (three samples). PCR sensitivity was affected by inhibitors and by the amount of DNA in the clinical samples. Our results indicate that direct detection of M. bovis in lymph nodes by PCR may be a fast and useful tool for bovine tuberculosis epidemic management in the region.     

Detection of Encephalitozoon cuniculi in the feline cataractous lens

Purpose Identification of Encephalitozoon cuniculi (E. cuniculi) as a possible causative agent for cataracts and uveitis in cats. Methods Within a 12‐month study period, cats that were presented with focal anterior cortical or mature cataract and secondary uveitis underwent a complete ophthalmic examination, complete blood count, serum biochemistry, serologic tests for E. cuniculi and tests for feline immunodeficiency virus (FIV), feline infectious peritonitis (FIP), feline leukemia virus (FeLV) and Toxoplasma gondii (T. gondii). PCR for DNA detection of E. cuniculi and T. gondii as well as cytologic examination of aqueous humor after paracentesis and phacoemulsified lens material were also performed. In addition histopathologic examination of the resected anterior lens capsule and attached lens epithelial cells was performed. Serologic testing for antibodies against E. cuniculi was also performed in 100 ophthalmologically healthy cats. Results Eleven (19 eyes) European shorthair cats with a median age of 3.5 years were included. Nine of 11 cats had bilateral cataracts, with 12/19 eyes having focal anterior cortical cataracts and 7/19 eyes having mature cataracts. In 14/19 eyes anterior uveitis was present. All cats had a positive antibody titer (1:80–1:10 000) for E. cuniculi. Encephalitozoon cuniculi DNA was detected by PCR and sequencing in 18/19 lenses and in 10/19 aqueous samples. Five tentative positive results were detected by cytologic examination. Spores were detected in 15/19 samples of lens material with histopathologic staining. Only 2/100 ophthalmologically healthy cats showed a positive antibody titer for E. cuniculi. Conclusion Encephalitozoon cuniculi is a cause of focal anterior cortical cataract and anterior uveitis in cats.     

感染细胞与痘疹样本中山羊痘病毒PCR检测方法的建立

比较Trizol法和SDS-蛋白酶K法提取山羊痘病毒DNA后,根据山羊痘病毒P32基因设计引物,建立了能检测感染细胞培养物与组织病料的PCR方法,结果显示以SDS-蛋白酶K法提取的病毒DNA在含量与纯度上均明显高于Trizol法;该PCR方法对山羊痘标准毒株细胞感染物能扩增出特异性的DNA条带,最小检出量为40.625ng;且能检出山羊痘疫苗毒Y株、贵州现场分离毒LD株和QL株的细胞感染物以及山羊痘疹样本中病毒DNA,而对鸡痘疫苗毒株感染细胞、正常细胞及正常山羊皮肤均为阴性反应。这些结果表明,建立的PCR方法具有特异性强、可靠性好、灵敏度高等特点,可用于山羊痘的临床诊断。     

Prevalence of serum antibodies to Toxoplasma gondii in free-ranging cats on Tokunoshima Island,Japan

The prevalence of Toxoplasma gondii infection in free-ranging cats on Tokunoshima Island was assessed by testing 125 serum samples using anti-T. gondii IgG indirect enzyme-linked immunosorbent assay. The overall seropositivity rate was 47.2% (59/125). Seropositivity rates in cats with body weight >2.0 kg (57.4%) were significantly higher than in those with body weight ≤2.0 kg (12.5%, P<0.01). Analysis of the number of seropositive cats by settlement revealed the presence of possibly-infected cats in 17 of 23 settlements, indicating the widespread prevalence of T. gondii on the island. This is the first study to show the seroprevalence of T. gondii in free-ranging cats on Tokunoshima Island. The information revealed in this paper will help to prevent the transmission of T. gondii among cats and also in both wild and domestic animals and humans on the island.     

Comparison of two DNA extractions and nested PCR, real-time PCR, a new commercial PCR assay, and bacterial culture for detection of Mycobacterium avium subsp. paratuberculosis in bovine feces.

In this study, 5 combinations of 2 DNA extractions and 3 polymerase chain reaction (PCR) techniques were compared with culture for the detection of Mycobacterium paratuberculosis directly from bovine feces. These combinations included a new commercial extraction technique combined with a commercial PCR/Southern blot technique, nested PCR (nPCR), or real-time PCR, and a university-developed extraction combined with nPCR or real-time PCR. Four of the 5 combinations had statistically similar sensitivities between 93% and 100% and specificity between 95% and 100%, when compared with culture results from 63 bovine fecal samples. These results indicated that using a commercial extraction with a commercial PCR/Southern blot, nPCR, or real-time PCR, or a university-developed extraction with real-time PCR would result in similar sensitivities to culture for the identification of M. paratuberculosis from bovine feces and are valid alternatives to culture.     

High rate of feline immunodeficiency virus infection in cats in the Brazilian semiarid region: Occurrence,associated factors and coinfection with Toxoplasma gondii and feline leukemia virus

To evaluate the occurrence of feline immunodeficiency virus (FIV) and factors associated with this and to demonstrate occurrences of coinfection with Toxoplasma gondii and feline leukemia virus (FeLV) in cats, a total of 103 blood samples were collected from owned cats, during home visits. To diagnose FIV and FeLV, immunochromatographic kit was used and serological diagnoses of T. gondii, the indirect immunofluorescence test was performed. The occurrence of FIV-seropositive cats was 23.3% (24/103) and the factor associated with infection was male sex. T. gondii seropositivity of 53.4% (55/103) was observed and 75% of FIV cases (18/24) were positive for T. gondii coinfection. Only 0.9% (1/103) was positive for FeLV. It can be concluded that the seroprevalence of FIV in cats in the Brazilian semiarid region is high and that FIV positive cats were also likely to be T. gondii seropositive, while FeLV had very low occurrence in the study region.     

Seroprevalence and risk factors associated with Toxoplasma gondii infection in pig farms from Catalonia, north-eastern Spain

Seroprevalence and associated risk factors for Toxoplasma gondii infection in pigs were analyzed in 1202 sera samples, including sows and pigs of three, seven, 11, 15 and 20 weeks of age, from 23 farms in Catalonia, north-eastern Spain. Antibodies were tested by the modified agglutination test (MAT) at titers ?1:25. Antibodies to T. gondii were found in 228 samples (19.0%; 95% CI: 16.8-21.2). The individual prevalence in animals higher than 7 weeks of age was 22.8% (174/762; 95% CI: 16.6-29.0) and the within-farm prevalence ranged from 7.1% to 36.4%. Statistically significant differences were found among age classes. The risk factors significantly associated with T. gondii seroprevalence were the presence of cats, percentage of mortality at weaning and the presence of outdoor facilities in the farms. The seroprevalence observed in the present study indicates widespread exposure to T. gondii among domestic pigs in Catalonia, which may have important implications for public health.