Undercooked Wild Boar Research Articles

Detection of Hepatitis E Virus in Game Meat (Wild Boar) Supply Chain in Umbria Region, Central Italy.

Consumption of raw or undercooked wild boar (WB) meat is considered an important risk factor for hepatitis E virus (HEV) infection in humans. The possibility of HEV contamination during the slaughtering practices may pose an additional risk. Based on these assumptions, we evaluated HEV contamination of WB meat hunted in Umbria (central Italy) during the 2022-2023 hunting season by real-time RT-PCR. Herein, we show that 10.8% of livers from slaughtered WB were positive for HEV RNA, thus providing an estimate of HEV infection in WB in the Umbria region. Then, by evaluating paired liver-muscle samples from both HEV-positive and HEV-negative animals, we found evidence of muscle HEV contamination in 33% and 14% of cases, respectively. This is the first report on the detection of HEV in WB meat in Umbria, an Italian region with diffuse WB hunting and consumption. The evidence of contamination provided by our study underscores the importance of adopting good hygienic practices in the processing stages of hunted WB carcasses to significantly reduce meat contamination and the risk posed for the final consumer.

High Seroprevalence of Toxoplasma gondii in Slovenian Wild Boars (Sus scrofa).

Simple SummaryToxoplasma gondii is a parasite that can cause serious disease in humans, especially in pregnant women. This parasite is found in many animals and people can be infected by eating raw or undercooked meat. Wild boar are known to harbor this parasite; populations and habitats have increased in the past decade, as it also increased the consumption of venison. The European population of wild boar has a mean prevalence of 26%. In our study, we found that the prevalence in the Slovenian wild boar population is 62%, the highest in the world to date. The prevalence was influenced by age and weight, but not by gender. In conclusion, the hunting community should be made aware of the high risk of parasite exposure when dealing with wild boar meat.Toxoplasma gondii is a zoonotic parasite of great public health concern. Wild boars could be considered an emerging source of toxoplasmosis in humans due to the popularity of venison and their increasing population. The aim of this study was to determine the seroprevalence of T. gondii in the Slovenian wild boar population and evaluate risk factors for human infection. Of 353 samples, 62% were positive for T. gondii using ELISA tests. This is the highest T. gondii seroprevalence reported to date in wild boar worldwide. The increase in prevalence with increasing age (p = 0.003) and weight (p = 0.002) were statistically significant, whereas gender was not (p = 0.781). Odds for being T. gondii-positive increased with age with the largest difference being between 2–3-year-old and 1–2-year-old animals (OR = 2.66, 95%CI: 1.03–6.85). Animals weighing 20–40 kg had a higher risk than animals weighing 0–20 kg (OR = 2.74, 95%CI: 1.21–6.20), whereas a further increase in the weight was not associated with increasing the odds. Due to the high Toxoplasma prevalence, the study concluded that the risk of exposure to T. gondii from handling raw or undercooked wild boar meat is high. Surveillance protocols should be established at the national level together with increased awareness within the hunting community.

Investigation of the Food-Transmitted Parasites Trichinella spp. and Alaria spp. in Wild Boars in Greece by Classical and Molecular Methods and Development of a Novel Real-Time PCR for Alaria spp. Detection.

Simple SummaryThere are many parasites that may be transmitted to humans via food, and meat is a major source of such infections. Trichinella spp. is one of the most important meat-transmitted parasites, while Alaria spp. may be considered an emerging pathogen, albeit to date rarely reported in humans. Raw and undercooked wild boar meat has been proven as a major source of human infection by both parasites. In the present study, an investigation of the presence of these parasites in wild boar meat was conducted for the first time in Greece. Classical parasitological methods and molecular techniques were implemented for the examination of samples collected from 128 hunted wild boars, and none of them were found positive for Trichinella spp. or Alaria spp. For the detection of Alaria spp., a novel molecular method was developed, offering a powerful complementary diagnostic tool that may be useful for the epizootiological surveillance of the parasite. The epizootiology/epidemiology, clinical implications, and importance of monitoring of these parasitic infections are briefly discussed.Foodborne parasitic diseases represent a major threat to public health. Trichinellosis, caused by the nematode parasite Trichinella spp., is one of the most important foodborne diseases, while alariosis, caused by the trematode parasite Alaria spp., is less common in humans, and rare cases have been reported only in the USA and Canada. Both parasites can infect humans via the consumption of raw or undercooked wild boar meat. In order to investigate the prevalence of these parasites in wild boar meat in Greece, samples from the diaphragm pillars and the region of the mandibular angle from 128 wild boars, hunted in Greece, were collected. The samples were examined by classical parasitological (compression, artificial digestion, and Alaria spp. migration) and by molecular (real-time PCR) methods. For Trichinella spp. an existent real-time PCR detecting all species likely to be present in Greece was applied, while for Alaria spp. a real-time PCR was developed, employing an LNA TaqMan probe targeting the large subunit ribosomal RNA gene. All examined wild boar samples from Greece resulted negative for Trichinella and Alaria species, indicating a low prevalence of infection in the examined population. The novel real-time PCR for Alaria spp. has 81.5% amplification efficiency and is able to detect 0.12 larvae per 50 g of tissue and could be utilized as a complementary to AMT diagnostic tool in surveillance.

Hepatitis E Virus Occurrence in Pigs Slaughtered in Italy.

Simple SummaryHepatitis E is now recognized as an emerging zoonotic disease in Europe caused by an RNA virus (HEV) and foodborne is the main route of transmission. Human cases have been linked to the consumption of contaminated pig liver sausages, raw venison, or undercooked wild boar meat. The zoonotic genotype HEV-3 is widespread in pigs at farm level but little information is available on the occurrence of HEV-positive pigs at the slaughterhouse. The aim of this study was to investigate the prevalence of HEV-positive pigs during slaughtering, to understand which biological samples (feces and organs) were more frequently HEV positive. Our results showed that pigs positive for HEV can be slaughtered and that the percentage of positive animals depends on the age of animals. The other main result is the presence of the virus in the plasma of animals, which may contribute to the contamination of meat (muscle). Nevertheless, muscles are rarely contaminated by HEV-RNA compared to liver, which is the organ of replication.In Europe, foodborne transmission has been clearly associated to sporadic cases and small clusters of hepatitis E in humans linked to the consumption of contaminated pig liver sausages, raw venison, or undercooked wild boar meat. In Europe, zoonotic HEV-genotype 3 strains are widespread in pig farms but little information is available on the prevalence of HEV positive pigs at slaughterhouse. In the present study, the prevalence of HEV-RNA positive pigs was assessed on 585 animals from 4 abattoirs located across Italy. Twenty-one pigs (3.6%) tested positive for HEV in either feces or liver by real-time RT-PCR. In these 21 pigs, eight diaphragm muscles resulted positive for HEV-RNA. Among animals collected in one abattoir, 4 out of 91 plasma tested positive for HEV-RNA. ELISA tests for the detection of total antibodies against HEV showed a high seroprevalence (76.8%), confirming the frequent exposure of pigs to the virus. The phylogenetic analyses conducted on sequences of both ORF1 and ORF2 fragments, shows the circulation of HEV-3c and of a novel unclassified subtype. This study provides information on HEV occurrence in pigs at the slaughterhouse, confirming that muscles are rarely contaminated by HEV-RNA compared to liver, which is the most frequently positive for HEV.

Geographical restriction of Hepatitis E virus circulation in wild boars (Sus scrofa) in Emilia-Romagna region, Northern Italy.

Hepatitis E virus (HEV) is a singlestrand RNA virus that causes an acute viral hepatitis in humans. Among its eight recognized genotypes, HEV-3 and HEV-4 are zoonotic, infecting humans, pigs and wild boars. Recently, HEV-3 has been also detected in red deer, which represents another reservoir of HEV. Consumption of raw pork products (mainly liver sausages), undercooked wild boar meat, raw wild boar liver and deer meat has been responsible for foodborne HEV human worldwide. From November 2018 to March 2019, liver samples collected from 97 wild boars hunted in Emilia-Romagna region (Northern Italy) were tested for HEV RNA. The hunting area included two territories for an extension of 33 km2, named A (about 13 km2,natural park, deciduous wood) and B (about 20 km2, cultivated fields in proximity of a river) areas. Distance between the two areas ranged between 8 to 10 km. A total of 73 wild boars were hunted in area A, and 24 in area B. HEV RNA was detected by Real-time RT– PCR in 23/73 liver samples of wild boars living in area A only (31.5% - 95% CI: 22.0-42.8%). The HEV sequences (n=13) clustered within genotype 3. The majority of positives belonged to animals < 12 months (12/25; 48%), followed by subadults (13-24 months) (7/16; 43.8%) and adults (4/32; 12.5%). This difference was found to be statistically significant (p=0.0024). In absence of pig farms, the restriction of HEV-positive animals to a well-defined territory of 13 km2 (Boschi di Carrega Regional Park) could hypothetically be related to the presence of red deer (Cervus elaphus), which lived in area A at the beginning of the hunting season. Further studies are needed to confirm or deny our hypothesis.

Detection of subtypes (3a, 4a and 4d) and high prevalence of hepatitis E virus in Korean wild boar

The hepatitis E virus (HEV) is known to have 4 genotypes but only one serotype. Genotype 1 and 2 infect humans only and genotype 3 and 4 infect humans, pigs and other animal species. Pig and wild boar are also known as reservoirs of HEV infection. Of the 2736 wild boars captured from 2011 to 2016 to investigate the HEV prevalence among Korean wild boars, 1041 serum samples were high seropositive (38.1%; 95% CI: 35.5–40.5) for HEV, which were detected using the anti-HEV antibody ELISA and the highest prevalence rate was 40.6% (684/1683) in 2016. Twenty four HEV strains were also identified from 1859 wild boar bloods captured between 2015 and 2016. The phylogenetic tree constructed based on the partial ORF2 gene revealed that the 23 Korean wild boar HEV strains belonged to genotype 4 (4a and 4d) showing the nucleotide sequences identities 83.4–100 %. The one Korean wild boar HEV strain belonged to genotype 3, segregated into subgenotype 3a. This suggested that major circulating in Korean wild boars is genotype 4a whereas genotype 3a and -4d is minor. It is important to the human public health that HEV with wild boar have potential high risk factor for transmission to human due to eating culture of Korean people with undercooked wild boar gallbladder.

Toxoplasma gondii and Trichinella infections in wild boars (Sus scrofa) from Northeastern Patagonia, Argentina

Wild boar (Sus scrofa) was introduced in many countries of the world and is recognized as carrier of many infectious diseases. Wild game meat consumption is recognized as a source of transmission of Toxoplasma gondii and Trichinella spp. The aim of the present study was to evaluate the prevalence of antibodies to T. gondii and Trichinella spp. in free-range wild boars in Northeastern Argentine Patagonia. Between 2014 and 2018, 144 blood samples and 423 muscle samples from 423 carcasses were collected. To detect T. gondii IgG, 144 sera were processed by an immunofluorescent antibody test, and to detect anti-Trichinella IgG, 125 sera and 304 muscle juice samples were processed by ELISA. Detection of first stage larvae in muscle was performed by artificial digestion. A total of 423 wild boars muscle samples were negative to Trichinella spp. by artificial digestion. Antibodies to Trichinella spp. were detected in 2.4% (3/125) of serum samples and in 1.64% (5/304) of meat juice samples. Antibodies to T. gondii infection were detected in 12.5% (18/144) of the serum samples. This is the first study to reveal the presence of antibodies to T. gondii in wild boars from Argentina. The present results suggest that consumption of raw or undercooked wild boar meat could represent a potential source risk for toxoplasmosis in humans and that Trichinella spp. is infrequent and/or that it circulates in low burdens among wild boars in Northeastern Patagonia.

Molecular survey of HEV infection in wild boar population in Italy.

Hepatitis E virus (HEV) is an RNA virus causing an acute generally self-limited disease in humans. An increasing number of autochthonous cases linked to zoonotic transmission of HEV genotype 3 have been reported over the last 10years in Europe. Pigs and wild boars are considered the main reservoirs. The principal route of transmission in Europe is food-borne, linked by direct or indirect evidence to the consumption of raw or undercooked pork products and wild boar meat. In this study, we sampled 92 wild boar (Sus scrofa) livers during active surveillance in five municipalities in Central Italy throughout the hunting season 2016-2017. HEV RNA was detected in 52.2% of liver sampled with prevalence ranging from 0.0% to 65.7%. HEV-positive wild boars were detected in all but one area of hunting. Phylogenetic analysis showed that strains clustered within the two subtypes HEV-3c and HEV-3f and displayed a wide range of phylogenetic diversity. Several strains were circulating in the areas investigated; animals possibly belonging to the same family group hunted by the same team were infected with a unique strain (100% nucleotide identity). As wild animals are a proven source of HEV transmission to humans and pigs, the high prevalence observed (mean 52.2%) poses a question on the risk of consuming raw or undercooked wild boar meat, and thus, this subject deserves further investigations.

Wild boars meat as a potential source of human trichinellosis in Poland: current data.

Trichinellosis is an epidemiological problem with a global distribution. In Poland a substantial increase of the wild boar population has been observed since 2010, together with an increased incidence of trichinellosis after ingestion of raw or undercooked wild boar products containing Trichinella spp. larvae. However, the actual number of human cases remains particularly difficult to determine. The aim of the present study was to determine the current prevalence and spread of these parasites within wild boars. The diaphragm pillars and tongue from 833 wild boars were collected from 2010 to 2014, as well as one wild boar meat sausage known to be a source of infection. The samples were tested for Trichinella spp. using pepsin digestion. Recovered larvae were identified at species level by multiplex polymerase chain reaction (multiplex PCR). The overall prevalence in all examined samples was found to be 2.0% (17/833). Recovered larvae were identified as T. spiralis and T. britovi (9/18 and 5/18, respectively). T. spiralis larvae were isolated from the sausage. Mixed infection was confirmed only once. Three isolates were not identified. The results of our study confirm that the wild boar plays a key role in the maintenance of Trichinella nematodes through the sylvatic cycle.

Hepatitis E virus-related liver alterations and viral antigen localization in European wild boar (Sus scrofa)

Sixty-four sera and faecal samples from hunted wild boar were submitted to indirect ELISA and RT-PCR to detect hepatitis E virus (HEV) infection. Liver samples were used to characterize liver alterations associated with HEV infection. Thirty-six (56.2 %) sera scored positive for HEV antibodies while six (9.4 %) faecal samples were virus-positive. A higher seroprevalence was found in adults with no differences between genders. Histological lesions were subclinical and characterized by mild multifocal and periportal lymphoplasmacytic infiltration of mainly CD3+ lymphocytes. All six liver samples from HEV RT-PCR-positive subjects showed immunohistochemical cytoplasmic positivity for viral antigen in inflammatory foci. HEV infection had no apparent effect on wild boar body condition and biometric parameters. However, HEV infection in wild boar constitutes a threat for hunters, forest workers and consumers of undercooked wild boar meat or liver.

Gluteal Apsesi Olan Bir Hastada Rastlantısal Olarak Saptanan Trichinella: Bir Olgu Sunumu

Trichinosis is caused most commonly by the nematode Trichinella spiralis. In this paper we report a case of gluteal abscess that developed after a man ingested undercooked wild boar meat. The biopsy revealed a small calcified cyst encircling a coiled threadlike worm. The diagnosis was trichinosis superimposed by pyogenic bacterial infection. As gram-positive cocci were seen, empirical treatment with amoxicillin and clavulanic acid was given for two weeks. The man was given albendazole at 400 mg twice daily for 14 days to eliminate infection from the intestine. The gluteal abscess healed uneventfully after two weeks. To the best of our knowledge, trichinosis superimposed with pyogenic bacterial infection is rarely reported in the literature.

Seroprevalence of Toxoplasma gondii and Trichinella spiralis infections in wild boars (Sus scrofa) in Korea

Toxoplasma gondii and Trichinella spiralis are important zoonotic pathogens with worldwide distributions. In Korea, several outbreaks of human toxoplasmosis and trichinellosis due to the consumption of infected wild animals have been reported. The purpose of this study was to determine the seroprevalence of T. gondii and T. spiralis infections in wild boars killed in Korea from December 2009 to October 2011. A total of 521 wild boars hunted in eight provinces were examined for antibodies to T. gondii and T. spiralis by using commercial ELISA kits. Overall, 25.1% of serum samples from individual boars were seropositive for T. gondii and 1.7% were seropositive for T. spiralis. Seropositive for T. gondii was found in the boars in all the eight provinces investigated and for T. spiralis in four provinces. This is the first report on the seroprevalence of T. gondii and T. spiralis infections in wild boars in Korea. The consumption of undercooked wild boar meat may expose humans to a high risk of infection.

A nationwide survey of hepatitis E virus (HEV) infection in wild boars in Japan: identification of boar HEV strains of genotypes 3 and 4 and unrecognized genotypes

To investigate the nationwide prevalence of hepatitis E virus (HEV) infection and to characterize HEV genomes among Japanese wild boars (Sus scrofa leucomystax), 578 boars captured in 25 prefectures from 2003 to 2010 were studied. Anti-HEV IgG was detected in 8.1%, and HEV RNA in 3.3% of boars. Among the 19 boar HEV isolates obtained from infected boars, 14 isolates (74%) were classified as genotype 3, 4 isolates (21%) as genotype 4, and the remaining isolate (wbJOY_06) was distantly related to all known HEV isolates of genotypes 1-4, differing by 18.4-25.0% and 18.0-24.3% within the 412-nucleotide sequence of ORF1 and ORF2, respectively. A genotype 4 boar HEV isolate (wbJGF_08-1) obtained herein shared 98.6% identity over the entire genome with a human HEV isolate obtained from a patient who developed acute hepatitis after consuming undercooked wild boar meat, suggesting that wild boars are also reservoirs for genotype 4 HEV in humans.

Age-related Toxoplasma gondii seroprevalence in Dutch wild boar inconsistent with lifelong persistence of antibodies.

Toxoplasma gondii is an important zoonotic pathogen that is best known as a cause of abortion or abnormalities in the newborn after primary infection during pregnancy. Our aim was to determine the prevalence of T. gondii in wild boar to investigate the possible role of their meat in human infection and to get an indication of the environmental contamination with T. gondii. The presence of anti-T. gondii antibodies was determined by in-house ELISA in 509 wild boar shot in 2002/2003 and 464 wild boar shot in 2007. Most of the boar originated from the “Roerstreek” (n = 673) or the “Veluwe” (n = 241). A binormal mixture model was fitted to the log-transformed optical density values for wild boar up to 20 months old to estimate the optimal cut-off value (−0.685) and accompanying sensitivity (90.6%) and specificity (93.6%). The overall seroprevalence was estimated at 24.4% (95% CI: 21.1–27.7%). The prevalence did not show variation between sampling years or regions, indicating a stable and homogeneous infection pressure from the environment. The relation between age and seroprevalence was studied in two stages. Firstly, seroprevalence by age group was determined by fitting the binary mixture model to 200 animals per age category. The prevalence showed a steep increase until approximately 10 months of age but stabilized at approximately 35% thereafter. Secondly, we fitted the age-dependent seroprevalence data to several SIR-type models, with seropositives as infected (I) and seronegatives as either susceptible (S) or resistant (R). A model with a recovery rate (SIS) was superior to a model without a recovery rate (SI). This finding is not consistent with the traditional view of lifelong persistence of T. gondii infections. The high seroprevalence suggests that eating undercooked wild boar meat may pose a risk of infection with T. gondii.

Seroprevalence and molecular detection of hepatitis E virus in wild boar and red deer in The Netherlands

To date, sources of hepatitis E virus (HEV) in the Netherlands, including swine and wild boar, have been identified, but no direct attribution to Dutch hepatitis E cases have been demonstrated. Other animal sources may exist. To identify these species, HEV RNA detection by RT-PCR is required, but complicated. A preselection based on serology may be useful. Therefore, wildlife species were studied by serology and molecular methods. Using a species-independent double-antigen sandwich ELISA, HEV-specific antibodies were detected in sera from 12% of 1029 wild boar ( Sus scrofa scrofa), in 5% of 38 red deer ( Cervus elaphus) and in none of 8 studied roe deer ( Capreolus capreolus). Differences in background signals were observed between species and accounted for by fitting finite mixture distributions. HEV RNA was detected in 8% of 106 wild boars, in 15% of 39 red deer and in none of 8 roe deer. In conclusion, HEV was shown to be present in European red deer for the first time. This preselection based on species-independent serological assays may be beneficial to identify new potential animal reservoirs of HEV. The consumption of Dutch undercooked wild boar and red deer meat may lead to human exposure to HEV.