Trichinella Larvae Research Articles

Muscle distribution of sylvatic and domestic Trichinella larvae in production animals and wildlife

Only a few studies have compared the muscle distribution of the different Trichinella genotypes. In this study, data were obtained from a series of experimental infections in pigs, wild boars, foxes and horses, with the aim of evaluating the predilection sites of nine well-defined genotypes of Trichinella. Necropsy was performed at 5, 10, 20 and 40 weeks post inoculation. From all host species, corresponding muscles/muscle groups were examined by artificial digestion. In foxes where all Trichinella species established in high numbers, the encapsulating species were found primarily in the tongue, extremities and diaphragm, whereas the non-encapsulating species were found primarily in the diaphragm. In pigs and wild boars, only Trichinella spiralis, Trichinella pseudospiralis and Trichinella nelsoni showed extended persistency of muscle larvae (ML), but for all genotypes the tongue and the diaphragm were found to be predilection sites. This tendency was most obvious in light infections. In the horses, T. spiralis, Trichinella britovi, and T. pseudospiralis all established at high levels with predilection sites in the tongue, the masseter and the diaphragm. For all host species, high ML burdens appeared to be more evenly distributed with less obvious predilection than in light infections; predilection site muscles harbored a relatively higher percent of the larval burden in light infections than in heavy infections. This probably reflects increasing occupation of available muscle fibers as larger numbers of worms accumulate. Predilection sites appear to be influenced primarily by host species and secondarily by the age and level of infection.

Human trichinellosis due to Trichinella britovi in southern France after consumption of frozen wild boar meat

Six patients were infected with Trichinella britovi in southern France following consumption of frozen wild boar meat, which had been frozen at -35 degrees C for 7 days. Microscopic examination of a sample of frozen wild boar muscle revealed the presence of rare encapsulated Trichinella larvae, identified as T. britovi. People eating wild boar must follow individual prophylactic rules such as efficient cooking of meat (at least 65 degrees C at the core for 1 minute) as recommended by the International Commission on Trichinellosis, or freezing exceeding four weeks at -20 degrees C.

Studies on vertical transmission of Trichinella spp. in experimentally infected ferrets ( Mustela putorius furo), foxes ( Vulpes vulpes), pigs, guinea pigs and mice

Vertical transmission of Trichinella spiralis was evaluated in ferrets ( n = 21), foxes ( n = 11), pigs ( n = 12), guinea pigs ( n = 16), and mice ( n = 41). The placental barrier to be crossed by migratory Trichinella larvae varies structurally in different animal species. Ferrets and foxes have an endotheliochorial placenta structure, guinea pigs and mice a haemochorial, and pigs an epitheliochorial placenta. The non-encapsulating Trichinella pseudospiralis larvae have an extended muscle migration prior to entering a muscle cell. To evaluate if T. pseudospiralis was more likely to be transmitted to offspring, an additional group of foxes ( n = 11) infected with T. pseudospiralis was included. Two different dose levels were used for ferrets, pigs, guinea pigs, and mice. In pigs and guinea pigs, infection was given at different times of the gestation period. Vertical transmission, measured as recovery of muscle larvae in the offspring, was demonstrated in both ferrets groups, in all four guinea pig groups, and in the high dose mouse group, but not in any fox or pig groups.

Trichinella pseudospiralis foci in Sweden

In Sweden, the prevalence of Trichinella infection in domestic pigs has greatly decreased since the 1970s, with no reports in the past 4 years. However, infected wild animals continue to be found. The objective of the present study was to identify the species of Trichinella present in animals of Sweden, so as to contribute to the knowledge on the distribution area and hosts useful for the prevention and control of this zoonosis. In the period 1985–2003, Trichinella larvae were detected in the muscles of 81/1800 (4.5%) red foxes ( Vulpes vulpes), 1/6 (16.7%) arctic fox ( Alopex lagopus), 1/7 (14.3%) wolf ( Canis lupus), 10/200 (5.0%) lynxes ( Lynx lynx), 4/8000 (0.05%) wild boars ( Sus scrofa), and 27/66 × 10 6 (0.000041%) domestic pigs. All four Trichinella species previously found in Europe were detected ( Trichinella spiralis, T. nativa, T. britovi and T. pseudospiralis). The non-encapsulated species T. pseudospiralis was detected in three wild boars from Hölö (Stockholm area) and in one lynx from Frösö (Östersund area), suggesting that this species is widespread in Sweden. These findings are consistent with those of a study from Finland, both for the unexpected presence of T. pseudospiralis infection and the presence of the same four Trichinella species, suggesting that this epidemiological situation is present in the entire Scandinavian region. The widespread diffusion of T. pseudospiralis in the Scandinavian region is also important in terms of it potential impact on public health, given that human infection can occur and the difficulties to detect it by the trichinelloscopic examination.

Trichinella papuaein Saltwater Crocodiles (Crocodylus porosus) of Papua New Guinea

<i>Trichinella papuae</i>in Saltwater Crocodiles (<i>Crocodylus porosus</i>) of Papua New Guinea

A preliminary investigation on the infectivity of Trichinella larvae in traditional preparations of walrus meat.

This study evaluated the infectivity of Trichinella nativa in freshly frozen walrus meat and traditionally aged walrus meat (igunaq) associated with two human outbreaks of trichinellosis in the Canadian Arctic. Trichinella larvae recovered from walrus meat stored at -20 degrees C for up to 20 months remained infective for guinea pigs inoculated with 135 or 716 larval doses. However, none of the 4-5 and 10-month-old igunaq preparations contained infective T. nativa larvae as measured by bioassays using mice and guinea pigs at inoculation doses ranging from 6 to 500 larvae. This indicates that the degradation process that occurred in the field can be sufficient to either kill Trichinella larvae or render them non-infective for mice and guinea pigs. Further research is needed to evaluate the food safety risk of traditional walrus igunaq aged under different field conditions and storage times.

Rediscovery of Trichinella spiralis in red foxes ( Vulpes vulpes) in Ireland after 30 years of oblivion

Objective. To determine whether or not Ireland can be considered as Trichinella-free, after more than 30 years of no reported infections in domestic and sylvatic animals and in humans. Methods. Samples of muscle tissue from the tongue, masseter, and foreleg of red foxes ( Vulpes vulpes) were subjected to artificial digestion and to multiplex-PCR analysis for identifying Trichinella larvae at the species level. Results. Four of 454 examined foxes were positive for larvae (overall prevalence 0.9%). The positive foxes had been collected in Donegal County (one fox; prevalence of 7.7%), Cork County (two foxes; 3.1%), and Waterford County (one fox; 4.2%). All larvae were identified as Trichinella spiralis. Conclusions. The detection of Trichinella-positive foxes in Ireland is an example of how the lack of infections among domestic animals and humans does not suffice for establishing Trichinella-free status. The results also confirm that the sylvatic cycle can last for tens of years, independently of the existence of a domestic cycle.

Comparison of a Modified Digestion Assay with Trichinoscopy for the Detection of Trichinella Larvae in Pork

A pepsin-HCl digestion assay and two compressorium techniques (trichinoscopy) for the identification of swine muscle tissue containing low levels of Trichinella larvae were compared as part of the test validation process for quality assurance purposes. Compressoria read with a stereomicroscope detected more larvae (P < 0.0001, n = 57) and more tissues (P =0.0047, n = 57) than did compressoria read with a projection microscope (trichinoscope). The digestion assay evaluated was 3.2 times as likely as the best compressorium technique to identify a positive tissue when these procedures were used to test 1 g of infected muscle (P < 0.001; 95% confidence interval for the odds ratio, 2.0 to 5.4; n = 161 and n = 189, respectively). Detection by trichinoscopy improved as the number of larvae in tissues increased to >2 larvae per g, but trichinoscopy was less sensitive than the digestion assay regardless of the tissue larval load. These data indicate that the quality controlled digestion assay used in this study is more sensitive than trichinoscopic techniques in the detection of tissues containing low levels of Trichinella larvae.

Trichinellazimbabwensis n.sp. (Nematoda), a new non-encapsulated species from crocodiles ( Crocodylus niloticus) in Zimbabwe also infecting mammals

Since 1995, Trichinella larvae have been detected in 39.5% of farmed crocodiles ( Crocodylus niloticus) in Zimbabwe. Morphological, biological, biochemical and molecular studies carried out on one isolate from a farmed crocodile in 2001 support the conclusion that this parasite belongs to a new species, which has been named Trichinella zimbabwensis n.sp. This species, whose larvae are non-encapsulated in host muscles, infects both reptiles and mammals. The morphology of adults and larvae is similar to that of Trichinella papuae. Adults of T. zimbabwensis cross in both directions with adults of T. papuae (i.e. male of T. zimbabwensis per female of T. papuae and male of T. papuae per female of T. zimbabwensis), producing F1 offspring which produce very few and less viable F2 larvae. Muscle larvae of T. zimbabwensis, like those of T. papuae, do not infect birds. Three allozymes (of a total of 10) are diagnostic between T. zimbabwensis and T. papuae, and five are diagnostic between T. zimbabwensis and Trichinella pseudospiralis, the third non-encapsulated species. The percentage of the pairwise alignment identity between T. zimbabwensis and the other Trichinella species for the cytochrome oxidase subunit I gene, the large subunit ribosomal-DNA (mt-lsrDNA) gene and the expansion segment five, shows that T. zimbabwensis is more similar to the two non-encapsulated species T. papuae (91% for cytochrome oxidase I; 96% for mt-lsrDNA; and 88% for expansion segment five) and T. pseudospiralis (88% for cytochrome oxidase I; 90% for mt-lsrDNA; and 66–73% for expansion segment five) than to any of the encapsulated species (85–86% for cytochrome oxidase I; 88–89% for mt-lsrDNA; and 71–79% for expansion segment five). This is the first non-encapsulated species discovered in Africa. The finding of a new Trichinella species that infects both reptiles and mammals suggests that the origin of Trichinella parasites dates back further than previously believed and can contribute to understanding the phylogeny and the epidemiology of the genus Trichinella.

Hunting Practices Increase the Prevalence ofTrichinellaInfection in Wolves From European Russia

From 1998 to 2000, 184 animals (82 wolves, 29 red foxes, 55 mustelids, 5 raccoon dogs, and 13 domestic dogs), mainly shot by hunters in the Tvier and Smoliensk regions of northwest European Russia, were tested for Trichinella larvae; 98 animals (53.3%) were found to be positive. The highest prevalence was detected in wolf (97.5%). Trichinella nativa was the most common species detected (98%). The diet of wolves was investigated by examining the stomach contents of 62 animals (75.6% of the total number of wolves examined for Trichinella). It consisted mainly of dog (36.4% of the total number of occurrences of all food items, PFO) and moose (31.2 PFO); however, during the hunting seasons of 1998-1999 and 1999-2000, skinned wolf carcasses were left in the forest as bait (567 carcasses, about 18,000 kg). This very high prevalence of Trichinella infection, the highest ever detected in a natural population of carnivores, could be explained by carnivore-carnivore transmission, influenced by the hunting practices adopted in the study area.

Outbreaks of human trichinellosis caused by consumption of dog meat in China.

Dog meat has become an important source of Trichinella infection for humans in China. The first documented outbreak of human trichinellosis resulting from the consumption of dog meat occurred in China in 1974. Until 1999, the outbreaks with this source of infection occurred mostly in Northeast of China (81 cases in five outbreaks in Jilin and two in Liaoning), Beijing (six cases) and Henan provinces (two cases). The epidemiological surveys were performed in nine Provinces or Autonomous Regions of China among 19,662 dogs samples. Dogs trichinellosis prevalence ranged from 7% in Henan to 39.5% in Heilongjiang, with an overall prevalence of 21.1%. Based on random amplified polymorphic DNA fingerprint (RAPD), some dog isolates of Heilonjiang and Jilin provinces were recently identified as Trichinella nativa, suggesting that this parasite is widely distributed among dogs in Northeast of China, while Trichinella spiralis in swine appears to be a common parasite throughout China. Since the Trichinella larvae in dog meat is resistant to freezing, caution should be paid to the consumption of dog meat even if it had been frozen.

Diagnosis and epidemiology of Trichinella infections in wildlife in The Netherlands.

Trichinella infections in foxes and wild boars were studied to determine the prevalence of infection in wildlife in the Netherlands. Muscles of 429 forelegs of foxes (Vulpes vulpes) and diaphragms of 11 wild boars (Sus scrofa) were artificially digested. Single larvae of Trichinella were identified at species level using random amplified polymorphic DNA (RAPD-PCR). In addition, an ELISA using ES antigen was used to test serum samples of 458 wild boars. The prevalence of Trichinella in foxes ranged from 3.9% in the the eastern part of the country, 13.1% in the central part of the country and 1.3% in the most western part of the country. Trichinella larvae of foxes were identified as T. britovi. In most samples, identification of larvae did not show reproducible results. The serological prevalence of Trichinella infections in wild boars was 6.8%. Wild boar populations are located in the central and in the southern part of the country. Trichinella larvae of wild boar were identified as T. spiralis. These results show that two Trichinella species are involved in the epidemiology of trichinellosis among wildlife. On the basis of previous reports, the present results suggest that the prevalence of Trichinella infection in wildlife is increasing in the last 20 years.

High prevalence of Trichinella nativa infection in wolf (Canis lupus) populations of Tvier and Smoliensk regions of European Russia.

Domestic and sylvatic trichinellosis have frequently been documented in European regions of Russia, with the highest prevalence reported in wolves (Canis lupus). From 1998 to 2000, 75 carcasses of wolves shot by hunters were tested for Trichinella larvae, and 73 (97.3%) of them were found to be positive. This very high prevalence of infection, the highest ever detected in a natural population of carnivores, could be explained by the human impact on the natural ecosystem. In fact, the diet of wolves living in the region under study mainly consists of carcasses of dogs and wolves, which are left in the forest or used as bait by hunters.

Horse trichinellosis, an unresolved puzzle.

In spite of routine controls to detect Trichinella larvae in horse-meat, human infections due to horse-meat consumption continue to occur in France and Italy. The epidemiology of horse trichinellosis since its discovery in 1975 is outlined, addressing the possible modes of natural transmission to horses, the need to develop more sensitive methods for detecting Trichinella larvae in horses, and the economic impact of horse trichinellosis. Investigations of human outbreaks due to horse-meat consumption have implicated single cases of inadequate veterinary controls on horses imported from non-European Union countries. In particular, most cases of human infection have been attributed to horses imported from Eastern Europe, where pig trichinellosis is re-emerging and the main source of infection in horses.

Evaluation of two PCR-based techniques for molecular epidemiology in Finland, a high-endemic area with four sympatricTrichinellaspecies

Trichinella larvae collected from wildlife, domestic and synanthropic animals in Finland were identified to species by two molecular techniques: Random amplified polymorphic DNA (RAPD) polymerase chain reaction (PCR) and the recently described multiplex PCR. The RAPD-PCR was very sensitive to the sub-optimal preservation muscle larvae and resulting in weak and smeared bands on the gels for such material. However, the same samples yielded easily recognizable bands in the multiplex PCR; this latter technique is then recommended for epidemiological studies, especially when the preservation of the samples is sub-optimal. For larvae in good condition the unequivocal bands obtained by multiplex was the easiest identifiable. Four species of Trichinella were identified in the material: T. spiralis, T. nativa, T. britovi, and T. pseudospiralis. Trichinella britovi is a new record for Finland, and T. pseudospiralis is a new record for Northern Europe. Mixed infections between T. britovi and T. spiralis, T. nativa and T. spiralis, and between T. britovi and T. nativa were detected; this is the first record of a mixed infection between T. spiralis and T. nativa in a naturally infected host. Raccoon dogs were the only host species from which all of the four Trichinella species were detected. Trichinella spiralis was found in both domestic animals and wildlife, but none of the sylvatic Trichinella species were detected in domestic pig.

DNA-binding activity in the excretory-secretory products of Trichinella pseudospiralis (Nematoda: Trichinelloidea).

A novel DNA-binding peptide of Mr approximately 30 kDa was documented for the first time in the excretory-secretory (E-S) products of the infective-stage larvae of Trichinella pseudospiralis. Larvae recovered from muscles of infected mice were maintained for 48 h in DMEM medium. E-S products of worms extracted from the medium were analysed for DNA-binding activity by the electrophoretic mobility shift assay (EMSA). Multiple DNA-protein complexes were detected. A comparison of the Mr of proteins in the complexes indicated that they could bind to the target DNA as a dimer, tetramer or multiples of tetramers. Site selection and competition analysis showed that the binding has a low specificity. A (G/C-rich)-gap-(G/T-rich)-DNA sequence pattern was extracted from a pool of degenerate PCR fragments binding to the E-S products. Results of immunoprecipitation and electrophoretic mobility supershift assay confirmed the authenticity of the DNA-binding protein as an E-S product.

Prevalence of Trichinella in Muscles of Some Domestic and Wild Mammals in Lithuania and their Impact on the Organism

A survey of the research into trichinellosis among 13 species of sylvatic and domestic animals conducted in 1987–2000 in Lithuania is presented herein. Muscle samples of 66 wild predators representing seven species: wolf (Canis lupus), red fox (Vulpes vulpes), raccoon dog (Nyctereutes procyonoides), pine marten (Martes martes), stone marten (Martes foina), weasel (Mustela nivalis), European polecat (Putorius putorius) and 105 domestic carnivores were examined. The investigations also included 156 wild boars (Sus scrofa), 1,383 domestic pigs, 43 brown rats (Rattus norvegicus) and 21 black rats (R. rattus). The highest extensiveness of infection with encapsulated Trichinella larvae was established among red foxes (9/27), raccoon dogs (3/11) and pine martens (5/8). These parasites were also detected in the muscles of one wolf, one European polecat, two out of the 156 (1.3%) wild boars and four out of the 86 (4.6%) stray cats. Pathogenicity and virulence of Trichinella isolate extracted from the wild boar to ...

The occurrence and ecology of Trichinella in marine mammals

Trichinella in marine mammals has a circumpolar arctic distribution and a narrow range of host species. It is commonly found in polar bears (Ursus maritimus), and increasingly in walruses (Odobenus rosmarus) where it presents a significant zoonotic hazard. This has resulted in the implementation of food safety programs in some arctic communities to test harvested walrus meat for Trichinella larvae prior to consumption. Trichinella has been reported infrequently in bearded seals (Erignathus barbatus) and ringed seals (Phoca hispida), and was once observed in a Beluga whale (Delphinapterus leucas). Cannibalism is probably the most important factor in maintaining a Trichinella cycle in polar bears. Arctic carnivores such as polar bears, arctic foxes (Alopex lagopus) and domestic dogs (Canis familiaris) have a high prevalence of Trichinella infection and the carcasses of at least some of these animals are deposited in the ocean. Scavenging of these carcasses by walruses probably occurs, but may not account for the high prevalence the parasite seen in this host species. Predation, carrion feeding and cannibalism have been documented for walruses and a sylvatic cycle similar to that of bears may exist in walrus populations. Seals and whales are likely infected through infrequent exposure to infected carcasses, either directly by scavenging or indirectly by consuming amphipods or fish that have fed on infected carcasses. The inefficiency of this mechanism may account for the low prevalence of Trichinella in seals and whales. It is known that isolates from marine mammals are cold tolerant, and infectious for man, and have been identified as Trichinella nativa (T2). Molecular and other phylogenetic studies would be useful to facilitate studies on the inter-relationship of Trichinella cycles involving marine and terrestrial mammals in the arctic and subarctic, and in the investigation of human outbreaks of trichinellosis in these areas.

EXPERIMENTAL TRICHINELLOSIS IN REINDEER

Six female reindeer calves were inoculated intraruminally with various doses of Trichinella muscle larvae. Four calves were inoculated with T. nativa, receiving 15,000 (n = 1), 5,000 (1), and 2,500 (2) larvae each. Two calves were inoculated with 5,000 T. spiralis larvae each. Blood samples were collected twice per week for total white blood cell (WBC) and differential counts and for serology using enzyme-linked immunosorbent assay (ELISA) based on T. spiralis excretory-secretory antigen. On day 56, the calves were slaughtered and muscle samples were examined according to the standard digestion method for Trichinella larvae. Blood samples were also collected twice a week from 4 uninoculated, but otherwise similar, reindeer calves corralled separately. Both the total WBC and eosinophil counts of the inoculated animals were, on average, higher during the experimental period. All the inoculated calves seroconverted, showing an increase in the optical density (OD) in the ELISA starting between day 23 and day 27 postinoculation. Very few muscle larvae (<0.08 larvae/g [lpg]) were to be found from the animals inoculated with T. nativa, but about 4 and 6 lpg were recovered from the masseter muscles of those inoculated with T. spiralis.

Trichinella papuae n.sp. (Nematoda), a new non-encapsulated species from domestic and sylvatic swine of Papua New Guinea

Encapsulated and non-encapsulated species of the genus Trichinella are widespread in sylvatic animals in almost all zoogeographical regions. In sylvatic animals from Tasmania (Australian region), only the non-encapsulated species Trichinella pseudospiralis has been reported. Between 1988 and 1998, non-encapsulated larvae of Trichinella were detected in five domestic pigs and six wild boars from a remote area of Papua New Guinea. Morphological, biological, and molecular studies carried out on one strain isolated from a wild boar in 1997 suggest that these parasites belong to a new species, which has been named Trichinella papuae n.sp. This species can be identified by the morphology of muscle larvae, which lack a nurse cell in host muscles, and whose total length is one-third greater than that of the other non-encapsulated species, T. pseudospiralis. Adults of T. papuae do not cross with adults of the other species and genotypes. Muscle larvae of T. papuae are unable to infect birds, whereas those of T. pseudospiralis do. The expansion segment V of the large subunit of the ribosomal DNA differs from that of the other species and genotypes. All of these features allow for the easy identification of T. papuae, even in poorly equipped laboratories. The discovery and identification of a second non-encapsulated species in the Australian region strongly supports the existence of two evolutionary lines in the genus Trichinella, which differ in terms of the capacity of larvae to induce a modification of the muscle cell into a nurse cell.