Validation of a real-time PCR assay for the detection of African swine fever virus in fresh pork meat juice.

African swine fever virus (ASFV) is the etiological agent of African swine fever (ASF), a disease with detrimental effects on the health, welfare, and production of domestic and wild pigs. The ASF laboratory confirmation is based on the analysis of blood, serum and organ samples. However, testing these samples could not be always convenient, economically feasible or possible. This study describes the validation process of a PCR-based assay targeting a portion of p72 gene, used for the molecular detection of ASFV, from meat juice samples obtained from pigs succumbed to ASFV. More specifically, we investigated the capability of a real-time PCR assay to detect ASFV DNA in meat juices obtained from the diaphragmatic muscle along with the correspondent spleens of 55 ASFV-positive pigs and wild boars sampled from confirmed outbreaks in Romania and from 73 ASFV-negative and regularly slaughtered healthy pigs collected in the Abruzzo region (Italy). The test was able to detect viral DNA in both types of samples, with lower Ct values in spleens (mean=21.11, median=20.61) than meat juices (mean=23.08, median=22.40). However, distributions of Ct values were strongly correlated each other (R2= 0.83, P<0.001). Considering the distribution of the observed Ct values in the 55 positive meat juice samples, a 1:10 dilution would be able to detect 90 % of positive samples, whereas a 1:100 dilution would reduce the detectability to 78 % of more contaminated samples. As meat juice could be obtained easily from muscles and considering the potential use of this test on pooled samples, it could represent a tool to aid the investigation of ASFV spread.

Revealing the Prevalence of Toxoplasma in Sierra Morena's Wild Boar: An ELISA-Based Study Using Meat Juice.

This research work focused on the prevalence of Toxoplasma gondii in wild boar from the Sierra Morena region. We conducted an ELISA analysis using meat juice samples. A total of 892 samples from six hunting seasons (2013-2019) were collected from the provinces that constitute the Sierra Morena Mountain range. These samples were analyzed using the Pigtype® ELISA kit, specifically developed for detecting T. gondii in meat juice. The overall prevalence of T. gondii in Sierra Morena was 23.2%. The highest prevalences were observed in Córdoba (31.6%) and Jaén (25.9%). These provinces exhibit the highest density of wild boar as well as the greatest presence of the Iberian lynx (Lynx pardinus). Further in-depth studies are necessary, but it appears that the presence of wild felids and scavenger behavior may be associated with this observation.

Meat Juice and Oral Fluid as Alternatives to Serum for Aujeszky Disease Monitoring in Pigs.

Aujeszky Disease Virus (ADV) is a double-stranded DNA virus with a lipoprotein envelope. The natural hosts of the infection are Suidae, but the virus can infect many other mammals. The gold-standard method identified by the WOAH for the diagnosis of Aujeszky disease is the ELISA method. The objective of this study was to compare the performance of meat juice and oral fluid matrices using a commercial ELISA kit designed for serum. A total of 80 blood and oral fluid samples were collected from four pig farms selected for this study. Diaphragm muscle samples of about 100 g and blood samples were collected from 213 animals at the abattoir. These biological matrices were collected from the same animals and tested using a competitive ELISA kit to detect antibodies against ADV. The relative accuracy of the meat juice compared to that of the serum was 96.7% (95% CI: 93.3-98.7%), with 206 correct results out of 213. The relative accuracy of the oral fluid compared to that of the serum was 61.3% (95% CI: 49.7-71.9%), with 58 correct results out of 80. Meat juice has a better combination of sensitivity and specificity than oral fluid. The usage of meat juice in routine diagnostic examinations could be achieved after further investigations to optimize the procedure.

Comparison of diaphragm meat juice and muscle swab samples to spleen and spleen swab samples for the detection of African swine fever viral nucleic acid.

Use of meat juice and muscle swabs at slaughterhouses may provide an easy-to-collect sample for African swine fever (ASF) surveillance. Meat juice has been experimentally shown to be a reliable sample for the detection of ASF virus (ASFV). We compared the detection of ASFV nucleic acid from diaphragm meat juice, diaphragm muscle swab, spleen, and spleen swabs from pigs with signs of ASFV infection at slaughterhouses around Kampala, Uganda. Pigs with ≥2 clinical or pathology signs at the time of slaughter had a spleen sample, spleen swab, diaphragm muscle sample, and diaphragm muscle swab collected. Meat juice was collected from muscle samples through a freeze-thaw cycle. Each sample was tested individually, and 72 spleen, meat juice, and muscle swab sample pools of 4 negative and 1 positive sample were tested, as well. Standard operating procedures from the USDA-Foreign Animal Disease Diagnostic Laboratory for viral DNA extraction and real-time PCR (rtPCR) were used. Of the 493 pigs evaluated, we classified as positive 357 (72.4%) diaphragm meat juice samples, 218 (44.2%) diaphragm muscle swabs, 247 (50.1%) spleen samples, and 241 (48.9%) spleen swabs. All spleen sample pools were positive (72 of 72; 100%), as were 71 of 72 (98.6%) meat juice pools and 67 of 72 (93.1%) muscle swab pools. Meat juice samples provided a reliable sample type for the detection by rtPCR of ASFV in pigs with natural infections.

Immunoproteomic Analysis of Trichinella britovi Proteins Recognized by IgG Antibodies from Meat Juice of Carnivores Naturally Infected with T. britovi.

Infection with Trichinella nematodes elicits non-specific and specific immune responses; these depend on the dose of infection, the nematode, and the host species. Few studies have examined the presence of specific antibodies against Trichinella spp. in the meat juice of wild animals. The aims of the study were to determine the prevalence of antibodies against Trichinella spp. in meat juice and to identify the specific proteins reacting with the meat juice from free-living carnivores naturally infected with the parasite. Meat juice samples were taken from foxes, badgers, raccoon dogs, and martens and tested with indirect ELISA. Antibodies against Trichinella spp. were detected in 10% of foxes and 46% of raccoon dogs. The ELISA results were confirmed by immunoblot, which revealed different protein patterns in meat juice from red foxes, raccoon dogs, and badgers. The most frequently observed bands were sent for further analysis by liquid chromatography coupled with tandem mass spectrometry (LC-MS/MS) for the detection of Trichinella britovi immunogenic proteins. The results confirm the presence of proteins such as serine protease and heat shock proteins associated with Trichinella infection. These findings provide that meat juice is a useful matrix for proteomic analysis.

Detection of Porcine Cysticercosis in Meat Juice Samples from Infected Pigs.

BACKGROUND: Seroprevalence of porcine cysticercosis has been generally studied using enzyme-linked immunosorbent assays (ELISA) detecting either antigens or antibodies in sera. However, serum is not always readily available. The objective of this study was to assess the diagnostic potential of meat juice in detecting porcine cysticercosis using a cysticercosis antibody ELISA. METHODS: Sera and meat juice samples from 13 different organs/tissues were collected from nine pigs naturally infected with cysticercosis and from six uninfected pigs reared under hygienic conditions. The sensitivity of the cysticercosis antibody ELISA in detecting porcine cysticercosis in meat juice samples was compared to that in serum samples from the same pigs. RESULTS: Using sera, cysticercosis was detected in all nine pigs harbouring cysticerci, but not in those reared under hygienic conditions. The sensitivity of the ELISA was highest in meat juice extracted from the diaphragm (100%), heart (89%) and neck muscle (78%) of the nine infected pigs, whereas it varied between 0 and 44% in the other samples. CONCLUSION: To the best of our knowledge, this is the first study for T. solium cysticercosis serology to use meat juice. Our results show that meat juice from pig carcass organs or muscles is a promising diagnostic specimen for the detection of porcine cysticercosis. More studies including a large sample size of pigs with varying degrees of cysticercosis infection are needed to further prove this concept.

High incidence of cold-tolerant Clostridium frigoriphilum and C. algidicarnis in vacuum-packed beef on retail sale in Germany.

Sixty vacuum-packed beef samples retailed in Germany were investigated for the occurrence of cold-tolerant Clostridium spp. After a storage period at 4 °C for eight weeks, meat juice from all samples was processed for culturing, DNA extraction and SYBR green qPCR for Clostridium species. After that, a previously developed multiplex qPCR, sequence analysis of the 16S rRNA gene, and MALDI-TOF MS were applied in order to identify Clostridium spp. found in samples. Subsequently, 23 samples were found positive for C. frigoriphilum (n = 19), C. estertheticum (n = 2), C. tagluense (n = 1) and C. lacusfryxellense/C. frigoris (n = 1). By using a new multiplex qPCR and a new RFLP method developed in this study, a further 15 meat juice samples were revealed to be contaminated with C. algidicarnis. With some samples being co-contaminated with two different species, 53% (n = 32) of all investigated vacuum-packed beef samples were found to be positive for cold-tolerant clostridia. This is the first report of detection and identification of C. algidicarnis in meat samples in Germany and Central Europe.

Seroprevalence of Toxoplasma gondii infection in sows and finishers from conventional and organic herds in Denmark: Implications for potential future serological surveillance.

Pigs are one of several host species for Toxoplasma gondii parasites, and consumption of infected pork may lead to toxoplasmosis in humans. We estimated seroprevalence in sows and finishers from conventional and organic herds in Denmark and discussed the strategies for reducing the risk from pork. We collected 447 blood samples from 59 herds, and additional meat-juice samples from 212 of the same pigs. Using a T. gondii IgG commercial ELISA test, we found 2% (95% CI = 0.4%-5%) apparent seroprevalence of T. gondii in conventional finishers, 11% (95% CI = 6%-17%) in organic finishers, 19% (95% CI = 11%-30%) in conventional sows and 60% (95% CI = 47%-72%) in organic sows. The odds of an animal testing positive for T. gondii was 16 times higher (95% CI = 4.6-74.3) in organic compared to conventional herds. The odds were 22 times higher (95% CI = 6.5-88.3) if the animal was a sow compared to a finisher. Meat-juice ELISA values were significantly correlated with plasma results (P < 0.001), but on average 64% of the blood-plasma ELISA values. Lowering the recommended cut-off from 20 to 13 percent positive values of the positive control for meat-juice ELISA, resulted in the meat-juice ELISA identifying 93% of the plasma positives as positive and 99% of the plasma negatives as negative. The time taken to detect one or more infected pigs from a T. gondii positive herd at slaughter was estimated using abattoir data on pigs (17,195,996) and batches (165,569) delivered to Danish abattoirs in 2018. The time to detection was affected by the seroprevalence, frequency at which the pigs were delivered, the number of samples tested per batch delivery and the batch sizes. Time to detection was long in conventional finisher herds due to low prevalence, and in sow herds because of intermittent delivery of a low number of sows. In organic finisher herds, time to detection was short due to medium prevalence and frequent delivery of a high number of finishers. Conventional finisher herds may be classified as low-risk, organic finisher herds as medium-risk, and conventional and organic sow herds as high-risk herds. Risk-mitigation strategies at processing plants (freezing or curing) or at the consumer level (heat treatment) for meat originating from high-risk herds, surveillance of medium-risk herds, and auditing for controlled housing (high biosecurity) in low-risk herds may be cost-effective alternatives to serological surveillance of all Danish pig herds.

Seroprevalence and Risk Factors of Toxoplasma gondii in Ruminant Meats from Wet Markets in Klang Valley and Abattoirs in Selangor, Malaysia.

(1) Background: The objective of this study was to determine the prevalence of T. gondii in meats of cattle, goat and sheep from wet markets in Klang Valley, and abattoirs in Selangor, Malaysia; (2) Methods: A total of 192 meat samples were purchased from 51 wet markets in six districts in Klang Valley (Gombak, Klang, Kuala Lumpur, Hulu Langat, Petaling and Putrajaya). Meanwhile, a total of 200 diaphragm samples were collected from two government abattoirs located in Shah Alam and Banting, Selangor. All meat juices from samples were subjected to an indirect-ELISA kit for the presence of T. gondii IgG antibodies. Furthermore, all 184 meat samples of goat and sheep were subjected to conventional nested PCR (B1 genes) for the detection of T. gondii DNA; (3) Results: T. gondii antibodies were detected in 25% (n = 98/392) of the samples with seroprevalence of 9.1% (19/208, CI: 5.9%-13.8%) in cattle meat; 54.7% (41/75, 95% CI: 43.5%-65.4%) in goat meat and 34.9% (38/109, CI: 26.6%-44.2%) in sheep meat. No T. gondii DNA was detected in any of the meat samples of goat and sheep. T. gondii seropositivity in wet market samples was higher in goat (OR = 37.1 CI 12.4-110.3) and sheep meat (OR 9.03 CI: 3.28-24.8) compared to cattle meat (OR = 1.0) At univariate level, meat from non-licensed abattoirs (OR = 6.0 CI: 2.9-12.3) and female animals (OR = 6.7; CI 1.9-22.6) had higher risks of being seropositive for T. gondii antibodies than licensed abattoirs and male animals, respectively. (4) Conclusions: This is the first report of seroprevalence of T. gondii in ruminant meats for human consumption in Malaysia. The findings signified high exposure of meat samples from wet markets to T. gondii and the need for control measures to reduce the likelihood of infection when such raw or undercooked meats are consumed.

Noninvasive strategies for surveillance of swine viral diseases: a review.

In view of the intensive development of the swine industry, monitoring and surveillance of infectious diseases require low-cost, effective, and representative population sampling methods. We present herein the state of knowledge, to date, in the use of alternative strategies in the monitoring of swine health. Blood sampling, the most commonly used method in veterinary medicine to obtain samples for monitoring swine health, is labor-intensive and expensive, which has resulted in a search for alternative sampling strategies. Oral fluid (OF) is a good alternative to serum for pooled sample analysis, especially for low-prevalence pathogens. Detection of viral nucleic acids or antiviral antibodies in OF is used to detect numerous viruses in the swine population. Meat juice is used as an alternative to serum in serologic testing. Processing fluid obtained during processing of piglets (castration and tail-docking) may also be used to detect viruses. These matrices are simple, safe, cost-effective, and allow testing of many individuals at the same time. The latest methods, such as snout swabs and udder skin wipes, are also promising. These alternative samples are easy to acquire, and do not affect animal welfare negatively.

Detection of Foot-and-Mouth Disease Virus in Swine Meat Juice.

Foot-and-mouth disease virus (FMDV) is a highly contagious agent that impacts livestock industries worldwide, leading to significant financial loss. Its impact can be avoided or minimized if the virus is detected early. FMDV detection relies on vesicular fluid, epithelial tags, swabs, serum, and other sample types from live animals. These samples might not always be available, necessitating the use of alternative sample types. Meat juice (MJ), collected after freeze-thaw cycles of skeletal muscle, is a potential sample type for FMDV detection, especially when meat is illegally imported. We have performed experiments to evaluate the suitability of MJ for FMDV detection. MJ was collected from pigs that were experimentally infected with FMDV. Ribonucleic acid (RNA) was extracted from MJ, sera, oral swabs, and lymph nodes from the same animals and tested for FMDV by real-time reverse transcription polymerase chain reaction (rRT-PCR). MJ was also tested for FMDV antigen by Lateral Flow Immunoassay (LFI). FMDV RNA was detected in MJ by rRT-PCR starting at one day post infection (DPI) and as late as 21 DPI. In contrast, FMDV RNA was detected in sera at 1-7 DPI. Antigen was also detected in MJ at 1-9 DPI by LFI. Live virus was not isolated directly from MJ, but was recovered from the viral genome by transfection into susceptible cells. The data show that MJ is a good sample type for FMDV detection.

Detection of anti-viral antibodies from meat juice of wild boars.

Wild boars are a reservoir for many zoonotic pathogens and a good sentinel for surveillance of zoonotic viral infections, but collection of serum samples from wild boars in the field is sometimes difficult and requires special equipment and techniques. In this study, ELISA using meat juices extracted from the heart and diaphragm of wild boars, instead of serum samples, was performed to detect antibodies against zoonotic pathogens, Japanese encephalitis virus and hepatitis E virus. The results of ELISA using meat juice samples were significantly correlated with those using serum samples and meat juice contained one-fifth the antibodies of serum samples. As meat juice is easily collected from wild animals in the field without special equipment and techniques, ELISA using meat juice is a simple and superior method for serological survey of zoonosis among wild animals.

Detection of porcine epidemic diarrhea virus (PEDV) IgG and IgA in muscle tissue exudate ("meat juice") specimens.

The diagnostic performance of porcine epidemic diarrhea virus (PEDV) IgG and IgA ELISAs was evaluated using paired serum and meat juice samples collected from PEDV-negative (n = 50) and PEDV-inoculated pigs (n = 87). Serum samples were tested by PEDV (IgG, IgA) ELISAs using a procedure performed routinely at the Iowa State University-Veterinary Diagnostic Laboratory (ISU-VDL). Serum samples were tested using PEDV serum IgG and IgA ELISA procedures as routinely performed at the Iowa State University-Veterinary Diagnostic Laboratory (ISU-VDL). Serum samples were diluted 1:50 and conjugate concentrations were 1/20,000 for IgG and 1/3000 for IgA. Meat juice samples were tested using the serum PEDV IgG and IgA ELISAs, with modifications, i.e., meat juice samples were diluted 1:25 and conjugate concentrations were 1/40,000 for IgG and 1/10,000 for IgA. Receiver operator characteristic (ROC) curve analyses were used to estimate diagnostic sensitivities and specificities over a range of sample-to-positive (S/P) cutoffs. Consistent with previous reports, this study showed that the PEDV IgG and IgA meat juice ELISAs provided excellent diagnostic performance and suggest that meat juice recovered from samples collected at slaughter could be used in routine PEDV surveillance.

Toxoplasma gondii infections in chickens - performance of various antibody detection techniques in serum and meat juice relative to bioassay and DNA detection methods.

Chickens, especially if free-range, are frequently exposed to Toxoplasma gondii, and may represent an important reservoir for T. gondii. Poultry products may pose a risk to humans, when consumed undercooked. In addition, chickens are regarded as sensitive indicators for environmental contamination with T. gondii oocysts and have been used as sentinels. The aim of the present study was to determine the suitability of commonly used antibody detection methods, i.e. the modified agglutination test (MAT), IFAT and ELISA to detect T. gondii-infected chickens. Samples of experimentally and naturally infected chickens were used. The infection state of all chickens was determined by Magnetic-Capture (MC-) real-time PCR (RT PCR). Naturally exposed chickens were additionally examined by mouse bioassay and conventional RT PCR on acidic pepsin digests (PD-RT PCR). Blood serum and meat juice of various sources were tested for antibodies to T. gondii. In naturally infected chickens, there was substantial agreement between the mouse bioassay and MC-RT PCR or the mouse bioassay and conventional PD-RT PCR. PD-RT PCR was slightly more sensitive than MC-RT PCR, as all (26/26) bioassay-positive chickens also tested positive in at least one of the tissues tested (heart, drumstick). By MC-RT PCR, 92.3% (24/26) of the naturally infected bioassay-positive chickens were positive. The diagnostic sensitivity of MC-RT PCR was clearly related to the organ examined. Based on a quantitative assessment of the MC-RT PCR results in experimentally infected chickens, brain and heart tissues harbored an at least 100 times higher parasite concentration than breast, thigh or drumstick musculature. In naturally infected chickens, only three out of 24 birds, which were MC-RT PCR-positive in heart samples, also tested positive in drumstick musculature. Under experimental conditions, the agreement between MC-RT PCR and the serological techniques revealed 100% diagnostic sensitivity and specificity. Under field conditions, examinations of sera by ELISA, IFAT and MAT showed good performance in identifying chickens that were positive in either a mouse bioassay, MC-RT PCR, or PD-RT PCR as illustrated by diagnostic sensitivities of 87.5%, 87.5% and 65.2%, respectively, and diagnostic specificities of 86.2%, 82.8% and 100%, respectively. The examination of meat juice samples from breast, drumstick or heart musculature revealed similar or even better results in the ELISA. The results in the MAT with meat juice from breast musculature were less consistent than those of ELISA and IFAT because a number of negative chickens tested false-positive in the MAT. The MAT performed similar to ELISA and IFAT when applied to test meat juice samples collected from heart, thigh or drumstick musculature.

Toxoplasma Gondii and Pre-treatment Protocols for Polymerase Chain Reaction Analysis of Milk Samples: A Field Trial in Sheep from Southern Italy.

Toxoplasmosis is a zoonotic disease caused by the protozoan Toxoplasma gondii. Ingestion of raw milk has been suggested as a risk for transmission to humans. Here the authors evaluated pre-treatment protocols for DNA extraction on T. gondii tachyzoite-spiked sheep milk with the aim of identifying the method that resulted in the most rapid and reliable polymerase chain reaction (PCR) positivity. This protocol was then used to analyse milk samples from sheep of three different farms in Southern Italy, including real time PCR for DNA quantification and PCR-restriction fragment length polymorphism for genotyping. The pre-treatment protocol using ethylenediaminetetraacetic acid and Tris-HCl to remove casein gave the best results in the least amount of time compared to the others on spiked milk samples. One sample of 21 collected from sheep farms was positive on one-step PCR, real time PCR and resulted in a Type I genotype at one locus (SAG3). Milk usually contains a low number of tachyzoites and this could be a limiting factor for molecular identification. Our preliminary data has evaluated a rapid, cost-effective and sensitive protocol to treat milk before DNA extraction. The results of the present study also confirm the possibility of T. gondii transmission through consumption of raw milk and its unpasteurised derivatives.

Comparison of commercial ELISA tests for the detection of Toxoplasma antibodies in the meat juice of naturally infected pigs.

Toxoplasmosis is a globally distributed protozoal zoonosis. Pigs are considered an important reservoir of Toxoplasma gondii and pork a major infection source of human toxoplasmosis. ELISA methods are commonly used diagnostic tools for detecting Toxoplasma infections. They are also used for slaughterhouse-based serological monitoring of toxoplasmosis in pigs to identify positive farms. The methods used are non-standardised with varying sensitivity and specificity. In our study, four commercial ELISA tests for the detection of Toxoplasma antibodies in the meat juice of slaughter pigs were compared with a modified agglutination test (MAT) as a reference. The cut-off values of the ELISA tests provided by the manufacturer varied between 0.20 and 0.50, and clearly influenced prevalence. The sensitivity of tests I, II and III varied between 96.4 and 78.6. Sensitivity was unacceptably low (3.6) for test IV (cut-off=0.30). Tests I, II and III had the highest accuracy and the best agreement with the reference test when a cut-off of 0.30 was used. Test II and III showed very good agreement (K=0.92 and 0.84, respectively) with the MAT. A very strong correlation (Pearson correlation >0.89) was observed between the S/P values of tests I, II and III. Our results demonstrate that the test and cut-off value used influence the results of the apparent seroprevalence studies.

Meat Juice Serology for Toxoplasma Gondii Infection in Chickens.

Toxoplasma gondii is an important foodborne zoonosis. Free-range chickens are at particularly high risk of infection and are also excellent indicators of soil contamination by oocysts. In the present study, hearts of 77 free-range chickens were collected at slaughter. T. gondii meat juice enzyme-linked immunosorbent assay was performed with a commercial kit, following validation with positive controls, from experimentally infected chickens, and negative ones. Out of 77 samples, only 66 gave sufficient meat juice for serology. Of these, 24 (36.4%) were positive for T. gondii considering the 5*standard deviation values (calculated on the optical density of negative controls), while all the samples were negative considering sample/positive% values. Parasite-specific polymerase chain reaction was carried out on all samples obtained from heart tissue and none were positive for the presence of T. gondii DNA. Results would suggest that further study on the use of meat juice with a validated serological test to detect T. gondii in chickens could lead to widespread epidemiological studies in this important intermediate host. However, sample collection and test specificity require further evaluation.

Pasture is a risk factor for Toxoplasma gondii infection in fattening pigs.

As consumer awareness of animal welfare increases throughout Europe, housing of pigs in more animal-friendly systems is becoming more common. There is concern that these free-range and organic management systems increase the prevalence of zoonotic meat-borne pathogens, such as Toxoplasma gondii. In this study we compared the seroprevalence of T. gondii between commercial fattening pigs raised on conventional and on organic farms in Sweden. Furthermore, potential associations between presence of T. gondii antibodies and type of production, access to pasture, and geographical region were analysed. A significant difference in T. gondii seroprevalence was found between conventional (1%) and organic pigs (8%). The higher odds of seropositivity in organic production was attributed to pasture access specifically (OR=1.8 for a one-month increase in length of pasture exposure). This study shows that the prevalence of T. gondii in Swedish conventional pigs is low. However, as pigs with access to pasture are at higher risk of infection and because the demand for animal-friendly production systems is increasing, there is an obvious need to practically manage the higher T. gondii presence in products from pigs raised in organic systems with outdoor access.

Seroprevalence of Toxoplasma gondii and direct genotyping using minisequencing in free-range pigs in Burkina Faso.

BACKGROUND: Swine are a major source of meat for humans. As such, they can play an important role in the epidemiology of human toxoplasmosis. Therefore, we performed an epidemiological study to determine the prevalence and genotypes of Toxoplasma gondii in Burkina Fasan swine. METHODS: The prevalence of T. gondii infection was evaluated in a 3-month prospective study at the slaughterhouse of Bobo-Dioulasso, Burkina Faso. Anti-Toxoplasma IgG titers were determined on meat juices from pig diaphragms using a commercially available ELISA assay. The DNA was extracted from 25mg of heart biopsies of seropositive animals (IgG ≥50% of the control) and the presence of T. gondii DNA was detected using a quantitative PCR assay. Genotyping was performed directly on DNA from PCR-positive biopsies using high-resolution melting and minisequencing analyses of the repeated B1 gene. RESULTS: The prevalence of carcasses positive for anti-Toxoplasma IgG was 29% (87/300) with no difference according to sex and age in contrast to the village of origin (p=0.018). Of the 87 seropositive animals, two were PCR positive (parasitic load at 64 and 128 parasites/mg of heart biopsy). Two new genotypes belonging to Type II and Type III and different from the genotypes previously described using minisequencing were identified. CONCLUSION: Our study provides the first T. gondii seroprevalence data in Burkina Fasan swine. In addition, this direct typing method suggests diversity of the T. gondii genotypes circulating in domestic animals in Burkina Faso. This needs to be confirmed on a wider sampling of subjects.

Toxoplasma gondii Infection in Hunted Wild Boars (Sus scrofa): Heart Meat Juice as an Alternative Sample to Serum for the Detection of Antibodies.

Toxoplasmosis is a global zoonosis caused by the protozoan parasite Toxoplasma gondii. Detection of antibodies to T. gondii in serum samples from hunted animals may represent a key step for public health protection. It is also important to assess the circulation of this parasite in wild boar population. However, in hunted animals, collection of blood is not feasible and meat juice may represent an alternative sample. The purpose of the present study was to evaluate heart meat juice of hunted wild boars as an alternative sample for post-mortem detection of antibodies to T. gondii by modified agglutination test (MAT). The agreement beyond chance between results from meat juice assessed with Cohen's kappa coefficient revealed that the 1:20 meat juice dilution provided the highest agreement. McNemars's test further revealed 1:10 as the most suitable meat juice dilution, as the proportion of positive paired samples (serum and meat juice from the same animal) did not differ at this dilution. All together, these results suggest a reasonable accuracy of heart meat juice to detect antibodies to T. gondii by MAT and support it as an alternative sample in post-mortem analysis in hunted wild boars.