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As a result of the increase of game meat intended for human consumption through Europe, a plethora of food-borne diseases, including trichinellosis, may occur in consumers, posing a relevant public health threat. Thus, this study aims to a citizen science approach to monitor the occurrence of Trichinella spp. in wild boar meat intended for human consumption, evaluating the risk of infection for consumers. Following the European Regulation 2015/1375 ( laying down specific rules on official controls for Trichinella in meat ), from 2015 to 2021, hunters ( n  = 478) were involved to collect diaphragm pillar samples of wild boars from mainland southern Italy, which were tested for Trichinella spp. L 1 larvae via HCl-pepsin digestion and Multiplex PCR. Overall, 139,160 animals were collected (average of 19,880 per year), being 14 (i.e., 0.01%) tested positive to Trichinella britovi by the combined biochemical and molecular approach. An average larval burden of 28.4 L 1 per gram of meat was found (minimum 3.2 - maximum 132.6). A statistically significant difference was found in the prevalence according to hunting seasons ( p  < 0.01, with higher values in 2016 and 2021) and regions of the study area ( p  < 0.01). No statistically significant decrease in the prevalence of T. britovi throughout the study period was found ( p  = 0.51), except in Apulia region ( p  < 0.01). These findings revealed a stable prevalence of T. britovi in wild boar meat intended for human consumption, suggesting a risk of infection for consumers, especially hunters and local markets users. Citizen science surveillance models could be promoted to improve trichinellosis control and prevention in a One Health perspective., Competing Interests: None., (© 2023 The Authors. Published by Elsevier B.V.)

Background: Domesticated pigs are the main source of Trichinella sp. infections for humans, particularly when reared in backyards or free-ranging. In temperate areas of southern Europe, most pigs are farmed under controlled housing conditions, but sows and sometimes fattening pigs have access to outdoors to improve animal welfare. The aim of the present study was to investigate whether outdoor access of breeding pigs farmed under controlled housing conditions can represent a risk for Trichinella sp. transmission when the farm is located in an agricultural area interspersed with wooded areas and badlands, where Trichinella spp. could be present in wildlife., Methods: Serum samples were collected from 63 breeding sows and one boar before and after their access to an open fenced area for 2 months and from 84 pigs that never had outdoor access. Samples were screened for anti-Trichinella antibodies by ELISA, and positive sera were confirmed using Western blot (Wb) excretory/secretory antigens. To detect Trichinella sp. larvae, muscle tissues from serologically positive and negative pigs were tested by artificial digestion., Results: Thirteen (20.6%) sows and one boar tested positive with both ELISA and Wb. No larvae were detected in muscle samples of serologically positive and serologically negative pigs. Positive serum samples were then tested by Wb using crude worm extract as antigens. The Wb banding pattern displayed was that characteristic of encapsulated species (Trichinella spiralis or Trichinella britovi)., Conclusions: The detection of anti-Trichinella antibodies without larvae in the pig muscles, supported by epidemiological data, suggests that pigs may have been exposed to T. britovi. This study stresses the importance of instigating monitoring systems at farm level to prevent Trichinella sp. transmission and to investigate, through a landscape parasitological study, the suitability of a site before the planting of a high containment level pig farm in which the sows can have outside access to improve their welfare during pregnancy., (© 2021. The Author(s).)

Within the genus Trichinella, Trichinella pseudospiralis is the only recognized non-encapsulated species known to infect mammals and birds. In October 2020, larvae recovered from muscle tissues of a wolf ( Canis lupus italicus ) originating from Molise Region, Central Italy, were molecularly confirmed as those of Trichinella britovi and T. pseudospiralis . This is the first detection of T. pseudospiralis from a wolf. In Italy, this zoonotic nematode was detected in a red fox ( Vulpes vulpes ), three birds ( Strix aluco , Athene noctua , Milvus milvus ) and five wild boars ( Sus scrofa ), and was also identified as the etiological agent of a human outbreak of trichinellosis in 2015. Since T. pseudospiralis is rarely reported from carnivore mammals in comparison to the encapsulated species frequently detected in these hosts, this finding opens the question of the role of carnivores as reservoirs for this parasite., Competing Interests: None.

Background: Domesticated and wild swine play an important role as reservoir hosts of Trichinella spp. and a source of infection for humans. Little is known about the survival of Trichinella larvae in muscles and the duration of anti-Trichinella antibodies in pigs with long-lasting infections., Methods: Sixty pigs were divided into three groups of 20 animals and infected with 10,000 larvae of Trichinella spiralis, Trichinella britovi or Trichinella pseudospiralis. Four pigs from each group were sacrificed at 2, 6, 12, 18 and 24 months post-infection (p.i.) and the number of larvae per gram (LPG) of muscles was calculated. Serum samples were tested by ELISA and western blot using excretory/secretory (ES) and crude antigens., Results: Trichinella spiralis showed the highest infectivity and immunogenicity in pigs and larvae survived in pig muscles for up to 2 years p.i. In these pigs, the IgG level significantly increased at 30 days p.i. and reached a peak at about 60 days p.i., remaining stable until the end of the experiment. In T. britovi-infected pigs, LPG was about 70 times lower than for T. spiralis at 2 months p.i. and only very few infecting larvae were detected at 6 months p.i., whereas no larvae were detected at 12, 18 and 24 months p.i. At 6 months p.i., degenerated/calcified larvae and cysts were detected in the muscles by trichinoscopy and histology. The IgG pattern showed by T. britovi-infected pigs was similar to that of T. spiralis-infected pigs, although seroconversion occurred some days later. The larval burden of T. pseudospiralis was slightly greater than for T. britovi at 2 months p.i., but no larvae were detected at 6 and 12 months p.i. In T. pseudospiralis-infected pigs, seroconversion occurred slowly, as in T. britovi-infected pigs. The IgG level showed a significant drop at 6 months p.i. and declining to the cut-off value at 12 months p.i., Conclusions: The longer survival of T. spiralis in pigs in comparison with the other two species highlights its exceptional dissemination potential. These results provide an explanation of the controversial data collected by parasitological and serological tools in the course of epidemiological investigations.

Parasite nematodes of the genus Trichinella are transmitted from one host to another through the ingestion of larvae present in striated muscles. The longer the survival of muscle larvae in host carcasses, the higher the probability of being ingested by a scavenging host. Thereby, these nematodes developed an anaerobic metabolism favouring their survival in decaying tissues. In addition, muscle larvae of three taxa, namely Trichinella nativa , Trichinella britovi and Trichinella T6, can survive freezing for several months to several years depending on the taxon. The aim of the present work was to investigate the survival time of T. britovi larvae in naturally infected host carcasses preserved beneath or above the snow. Fox and raccoon dog carcasses naturally infected with T. britovi larvae were preserved beneath or above the snow in a cold mountainous area. Temperature and relative humidity were recorded. Every 14 days, muscle samples collected from each carcass, were digested and larvae were counted and given per os to laboratory mice to evaluate their reproductive capacity index (RCI). The RCI of larvae in carcasses preserved beneath the snow (the subnivium) ranged from 23 to 25 at day 0, to 12-18 after 112 days. In contrast, the RCI of larvae in carcasses preserved above the snow ranged from 22 to 27 at day 0, to 0.0 after 112 days. The difference between the RCIs of larvae beneath the snow and above the snow was statistically significant ( P  < 0.01). These data corroborate the hypothesis that the subnivium with its environmental stability favours the survival of Trichinella larvae in host muscles, increasing the probability of their transmission to other hosts. On the other hand, the environment above the snow, characterized by sudden temperature variations, causes strong environmental stress for larvae in host carrions causing their death.

Background: In Europe, the life-cycle of Echinococcus multilocularis is predominantly sylvatic, involving red foxes (Vulpes vulpes) as the main definitive hosts and rodents such as muskrats and arvicolids as intermediate hosts. The parasite is the etiological agent of human alveolar echinococcosis, a malignant zoonotic disease caused by the accidental ingestion of eggs shed by definitive hosts in their faeces. The aims of this study were to investigate the prevalence of E. multilocularis in red foxes and raccoon dogs (Nyctereutes procyonoides) and to study the environmental factors favouring the perpetuation of the parasite in Latvia., Methods: A total of 538 red foxes and 407 raccoon dogs were collected across Latvia from 2010 to 2015. The sedimentation and counting technique was used for collecting E. multilocularis adult worms from fox and raccoon dog intestines. The morphological identification of the parasite was confirmed by molecular analysis., Results: The prevalence of E. multilocularis was significantly higher in foxes (17.1%; intensity of infection 1-7,050 worms) (P < 0.001) than in raccoon dogs (8.1%; intensity of infection 5-815 worms). In foxes, a significant positive correlation (r (10)  = 0.7952, P = 0.001) was found between parasite prevalence and the intensity of infection. A positive relationship (R s  = 0.900, n = 5, P = 0.037) between parasite prevalence and precipitation was also observed. In raccoon dogs, a significant negative relationship (F (1,8)  = 9.412, P = 0.015) between animal density and parasite prevalence, and a significant positive relationship (F (1,8)  = 7.869, P = 0.023) between parasite prevalence and agricultural land cover, were detected., Conclusions: The results of this study confirm the red fox as the most important definitive host of E. multilocularis and, consequently, as the main target for control programmes in the Baltic countries. Raccoon dogs seem to play a secondary role in the life-cycle of E. multilocularis within the investigated European region.

Background: Parasites of the genus Trichinella are zoonotic nematodes common in carnivores throughout the world. We determined the prevalence and species of Trichinella infections in Florida panthers (Puma concolor coryi)., Methods: Tongues from Florida panthers were collected at necropsy and examined by pepsin-HCl artificial digestion for infection with Trichinella spp. DNA was extracted from larvae and multiplex PCR using Trichinella species-specific primers was used to genotype the worms., Results: Trichinella spp. larvae were detected in 24 of 112 (21.4%; 14.6%-30.3%) panthers. Sixteen of the panthers (14.3%) were infected with T. pseudospiralis, 1 (0.9%) was infected with T. spiralis, and 2 (1.8%) had mixed infections of T. pseudospiralis and T. spiralis. Trichinella spp. larvae from 5 panthers were not identified at the species level due to degraded DNA., Conclusions: This is the highest prevalence of T. pseudospiralis detected in North America up to now and suggests the Florida panther is a key mammalian reservoir of this parasite in southern Florida. Trichinella pseudospiralis can infect both mammals and birds indicating the source of infection for Florida panthers could be broader than believed; however, birds represent a small percentage (0.01%) of the cat's diet. Since wild pigs (Sus scrofa) can be parasitized by both T. pseudospiralis and T. spiralis and these swine can comprise a large portion (~40%) of a panther's diet in Florida, we believe that Florida panthers acquired these zoonotic parasites from feeding on wild pigs.