Evaluation of the PrioCHECK™ Trichinella AAD kit to detect Trichinella spiralis, T. britovi, and T. pseudospiralis larvae in pork using the automated digestion method Trichomatic-35.

Trichinellosis is a potentially deadly parasitic zoonosis that is contracted by consuming undercooked infected meat. Reliable detection of infectious Trichinella spp. larvae in meat is therefore pivotal to ensure consumer's safety. The recently authorised PrioCHECK™ Trichinella Alternative Artificial Digestion (AAD) test kit appears promising when used with the standard magnetic stirrer method, but evaluation with other apparatus types is lacking. In this study, the performance of the AAD kit in an adapted Trichomatic-35 (TM35) instrument was evaluated, first, at the Swiss National Reference Laboratory for trichinellosis (NRL); second, in a ring trial involving four Swiss official laboratories. Proficiency pork samples spiked with larvae of Trichinella spiralis, T. britovi, or T. pseudospiralis were tested with the AAD kit and with the reference pepsin-HCl digestion method in TM35 instruments. At the NRL, both methods yielded identical qualitative and similar quantitative results independently of the Trichinella species. In the ring trial, satisfactory results were obtained for 47/50 (94.0%) (AAD) and 62/67 (92.5%) (reference method) of the analysed samples. Technical problems impairing analysis were more frequently observed with the AAD kit (n = 22) than with the reference method (n = 5) and were mainly (16/22) reported by one of the external labs. When no technical issues were recorded, the performance of both methods was comparable, in agreement with the observations at the NRL; however, these results suggest a need for further training with the kit and standardisation of the adapted TM35 instruments.

Experimental infection of tigerfish (Hydrocynus vittatus) and African sharp tooth catfish (Clarias gariepinus) with Trichinella zimbabwensis.

Trichinella zimbabwensis naturally infects a variety of reptilian and wild mammalian hosts in South Africa. Attempts have been made to experimentally infect piranha fish with T. zimbabwensis and T. papuae without success. Tigerfish (Hydrocynus vittatus) and African sharp tooth catfish (Clarias gariepinus) are accomplished predators cohabiting with Nile crocodiles (Crocodylus niloticus) and Nile monitor lizards (Varanus niloticus) in southern Africa and are natural hosts of T. zimbabwensis. To assess the infectivity of T. zimbabwensis to these two hosts, 24 African sharp tooth catfish (mean live weight 581.75 ± 249.71 g) randomly divided into 5 groups were experimentally infected with 1.0 ± 0.34 T. zimbabwensis larvae per gram (lpg) of fish. Forty-one tigerfish (mean live weight 298.6 ± 99.3 g) were randomly divided for three separate trials. An additional 7 tigerfish were assessed for the presence of natural infection as controls. Results showed no adult worms or larvae of T. zimbabwensis in the gastrointestinal tract and body cavities of catfish sacrificed at day 1, 2 and 7 post-infection (p.i.). Two tigerfish from one experimental group yielded 0.1 lpg and 0.02 lpg of muscle tissue at day 26 p.i. and 28 p.i., respectively. No adult worms or larvae were detected in the fish from the remaining groups sacrificed at day 7, 21, 28, 33 and 35 p.i. and from the control group. Results from this study suggest that tigerfish could sustain T. zimbabwensis under specific yet unknown circumstances.

Acute phase protein pattern and antibody response in pigs experimentally infected with a moderate dose of Trichinella spiralis, T. britovi, and T. pseudospiralis.

The aim of the present study was to evaluate the acute-phase protein (APP) response in three groups of pigs experimentally infected with a moderate infective dose, i.e. 1000 muscle larvae (ML) of Trichinella spiralis, 3000 ML of Trichinella britovi, and 2000 ML of Trichinella pseudospiralis. Over a 62-day period of infection, we examined the serum level and kinetics of the haptoglobin (Hp), C-reactive protein (CRP), serum amyloid A (SAA), and pig major acute-phase protein (pig-MAP). In addition, to better understand the immune response of pigs experimentally infected with three different species of Trichinella, the kinetics of IgG and IgM antibodies against excretory-secretory (ES) antigens of Trichinella ML were also investigated. In order to assess anti-Trichinella IgG dynamics, we used a commercial and an in-house ELISA based on both heterologous (T. spiralis) and homologous (T. spiralis, T. britovi, and T. pseudospiralis) Trichinella species ES antigens. Among the four APPs analyzed, the concentration of CRP and pig-MAP significantly increased only in T. britovi-infected swine when compared with control pigs. This took place as early as 6 days post-infection (dpi). Hp was the only APP whose concentration significantly increased in pigs infected with T. pseudospiralis, this occurring as late as on day 62 pi. Despite the statistical differences found, increases in pig-MAP, CRP, and Hp levels were rather mild and transitory; none of these proteins were found to be elevated in the serum of all experimental groups of pigs at the same time point after infection. Specific IgG antibodies against ES antigens of Trichinella ML were first detected by the commercial and in-house T. spiralis ML ES-antigen ELISAs on days 30, 36 and 36 pi in pigs experimentally infected with T. spiralis, T. britovi, and T. pseudospiralis, respectively. However, seroconversion in pigs experimentally infected with T. britovi was detected slightly earlier (30 dpi) when the ELISA based on homologous rather than heterologous ES antigens was applied. In serum samples from pigs infected with T. spiralis, statistically significant increases in the level of specific IgM antibodies against T. spiralis ML ES antigens were first detected on day 30 pi and after this time, their concentration began to decrease. No changes in the level of anti-Trichinella IgM were observed in T. britovi- or T. pseudospiralis-infected pigs throughout the entire period of the experiment.

The retention and concentration of glycogen in Trichinella nativа in the winter-spring period.

The metabolism of Trichinella spp. is primarily anoxybiotic in nature. Their main energy source is glycogen, which is stored in the stichocites at the muscular stage of the larval development. When subject to tow temperatures the Trichinella larvae consume glycogen and neutral fats to provide for basal metabolism until the energy supplies reach the critical level. The present study establishes the glycogen concentration as well the invasive activity of T. nativа when affected by low temperatures in natural conditions. The carcasses of infected laboratory rats were placed in containers beneath the snow cover, in the natural conditions of a game husbandry in Central Russia. The viability, invasive capacity and the glycogen level were monitored in the Trichinella larvae monthly. The invasive capacity of Trichinella larvae was established based on the presence of the larvae in the muscular tissue of laboratory mice after the peroral administration of the helminth larvae. On the 45 day of the experiment, the mice were euthanized by cervical dislocation, and if the Trichinella larvae could be discovered in the muscular tissue with the help of the trichinelloscopic compression method, the invasive capacity of the Trichinella larvae was viewed as positive. To establish the quantitative value of glycogen content in Trichinella larvae a modified method was used. In order to measure the glycogen level in the T. nativa larvae isolated by fermentation larvae were counted in one drop of the suspended sedimentation in the Migacheva-Kotelnikov chamber. To establish the quantitative value of glycogen content in Trichinella larvae a method based on the treatment of glycogen with iodine, optical density measurement with a refractometer MКMФ-02 was used. For the purpose of measuring the concentration of glycogen in Trichinella larvae in the suspended sedimentation a calibration curve was used. The studies showed that the viability indicator of the Trichinella larvae which had been preserved in natural conditions in the four months of the winter-spring period, in the muscular tissue of laboratory rats remained high (over 90 %). The glycogen concentration in one helminth larva was 0.041 µg in January, 0.033 µg in February, 0.015 µg in April. The invasive capability of the preserved Trichinella larvae was considerably reduced to 33.3 %. In the winter period, under temperatures below 0 °C, a decrease in the glycogen concentration in the Trichinella larvae was observed.

Comparison between Trichinella patagoniensis and Trichinella spiralis infection in BALB/c mice.

In Argentina, trichinellosis is an endemic disease acquired mainly through consumption of raw pork infected with nematodes larvae from the Trichinella genus. For years, the only species involved in outbreaks in humans and pig foci in Argentina was Trichinella spiralis. In 2008 the presence of a new Trichinella taxon from a cougar (Puma concolor) was detected and recorded in the province of Rio Negro, Argentina, and the finding was established as a new species in 2012: Trichinella patagoniensis. To the best of our knowledge, there is no information available on the intestinal phase and antibody response in a susceptible host during T. patagoniensis infection. Therefore, our research has been designed to study experimental infection with T. patagoniensis compared to infection with T. spiralis in BALB/c mice. One hundred and twenty eight BALB/c mice were divided into two groups and individuals in each group were infected per os with 500 larvae of T. patagoniensis or 500 larvae of T. spiralis, respectively. After that, they were euthanized on different days. Adult worm recovery from small intestines and artificial digestion of each carcass was performed. Histopathology of small intestines was performed using hematoxylin-eosin staining. Systemic cytokines and antibody kinetics were evaluated. Intestinal adult worm recovery of T. patagoniensis and T. spiralis took place until day 17 and 25, respectively. Systemic IFN-γ, IL-10, and TNF showed significant variations in T. patagoniensis infected mice. Seroconversion was detected in animals as from 15 days post-infection (pi) for both T. patagoniensis and T. spiralis, reaching the highest OD value at 42 days pi. Similar microscopic lesions were observed in the small intestine from mice infected with the same dose of T. spiralis and T. patagoniensis. Our findings contribute new information regarding the intestinal phase and the antibody kinetics of T. patagoniensis in BALB/c mice.

The Disease Ecology, Epidemiology, Clinical Manifestations, and Management of Trichinellosis Linked to Consumption of Wild Animal Meat.

Historically, human trichinellosis was caused by Trichinella spiralis and transmitted to humans by consumption of undercooked domestic pork. Today, most cases of trichinellosis are caused by other Trichinella species and transmitted by consumption of raw or undercooked wild game meats. Given the increasing global prevalence of wild animal meat-linked trichinellosis, the objectives of this review are: 1) to describe the life cycle and global distribution of Trichinella worms; 2) to describe the changing epidemiology of trichinellosis; 3) to describe the clinical phases of trichinellosis; 4) to recommend the latest diagnostic tests; and 5) to recommend treatment and prevention strategies. Internet search engines were queried with keywords as subject headings to meet the objectives of this review. Although trichinellosis surveillance systems and laws regulating commercial pork production have limited T spiralis-caused trichinellosis in Europe and the United States, trichinellosis due to consumption of raw and undercooked wild boar and feral hog meat continues to occur throughout Southeast Asia. Trichinellosis due to consumption of raw or undercooked meats of other infected game, such as bear, deer, moose, and walrus, continues to occur worldwide. Only adherence to hygienic practices when preparing wild game meats and cooking wild game meats to recommended internal temperatures can prevent transmission of trichinellosis to humans. Wilderness medicine clinicians should be prepared to advise hunters and the public on the risks of game meat-linked trichinellosis and on how to diagnose and treat trichinellosis to prevent fatal complications.

First detection of Trichinella pseudospiralis infection in raccoon (Procyon lotor) in Central Europe.

The raccoon (Procyon lotor) is a North American carnivore introduced to Europe in the 20th Century. Raccoons are believed to be the potential hosts of many parasites, or to be involved in their transmission to other animals. Nematodes of the genus Trichinella can infect many carnivorous and omnivorous animals worldwide. The aim of the present study was to determine the occurrence of Trichinella spp. infection in raccoons in Central Europe. Muscle samples were collected from various regions of Poland, the Czech Republic and Germany during the years 2012-2016. The larvae of Trichinella spp. were detected in 11 raccoons, and these were identified as T. spiralis and T. pseudospiralis by multiplex PCR (89.9% and 9.1%, respectively). No mixed infection was observed. This is the first report describing the occurrence of T. spiralis and T. pseudospiralis in P. lotor in Central Europe. Our findings also show that the raccoon population acts as a reservoir of Trichinella pseudospiralis.

Trichinella spp. biomass has increased in raccoon dogs (Nyctereutes procyonoides) and red foxes (Vulpes vulpes) in Estonia.

BACKGROUND: Raccoon dogs and red foxes are well-adapted hosts for Trichinella spp. The aims of this study were to estimate Trichinella infection prevalence and biomass and to investigate which Trichinella species circulated in these indicator hosts in Estonia. METHODS: From material collected for evaluating the effectiveness of oral vaccination program for rabies eradication in wildlife, samples from 113 raccoon dogs and 87 red foxes were included in this study. From each animal, 20 g of masseter muscle tissue was tested for the presence of Trichinella larvae using an artificial digestion method. The Trichinella larvae were identified to species level by multiplex polymerase chain reaction method. RESULTS: The majority of tested animals were infected with Trichinella spp. The parasite species identified were T. nativa and T. britovi. The apparent infection prevalence was 57.5% in raccoon dogs and 69.0% in red foxes, which were higher than previous estimates. In addition, the larval burden had also increased in both hosts. We estimated that in 2011-2012, the Trichinella spp. biomass was more than 15 times higher in raccoon dogs and almost two times higher in red foxes than in 1992-2000 (based on mean larval burden), and almost 20 times higher in raccoon dogs and almost five times higher in red foxes than in 2000-2002 (based on median larval burden). CONCLUSIONS: Raccoon dogs and red foxes are relevant reservoirs for Trichinella spp. in Estonia. The biomass of Trichinella circulating in sylvatic cycles was substantial and had increased: there is substantial infection pressure in the sylvatic cycle.

Quality Control of Trichinella Testing at the Slaughterhouse Laboratory: Evaluation of the Use of a 400-Micrometer-Mesh-Size Sieve in the Magnetic Stirrer Method.

The performance of a 400-µm-mesh-size sieve (sieve400) has not previously been compared with that of a 180-µm-mesh-size sieve (sieve180). Using pork samples spiked with 0 to 10 Trichinella muscle larvae and an artificial digestion method, sieve performance was evaluated for control of Trichinella in meat-producing animals. The use of a sieve400 resulted in 12% lower larval counts, 147% more debris, and 28% longer counting times compared with the use of a sieve180. Although no false-negative results were obtained, prolonged counting times with the sieve400 may have an impact on performance in a high-throughput environment such as a slaughterhouse laboratory. Based on our results, the sieve180 remains the sieve of choice for Trichinella control in meat in slaughterhouse laboratories, according to the European Union reference method (European Commission regulation 2075/2005). Furthermore, the results of the present study contribute to the discussion of harmonization of meat inspection requirements among countries.

Efficacy of maslinic acid and fenbendazole on muscle larvae of Trichinella zimbabwensis in laboratory rats.

Trichinellosis is a zoonotic disease caused by nematode species of the genus Trichinella. Anthelmintics targeting the intestinal adults and muscle-dwelling larvae of Trichinella spp. have been tested, with limited success. This study was aimed at determining the efficacy of maslinic acid and fenbendazole on muscle larvae of Trichinella zimbabwensis in laboratory rats. Forty-two Sprague-Dawley rats, with an average weight of 270 g and 180 g for males and females respectively, were infected with T. zimbabwensis larvae. Infected rats were randomly assigned to three groups which were subjected to single treatments with each of maslinic acid, fenbendazole and a combination of both on day 25 post-infection (pi), and three groups which were subjected to double treatments with each of these drugs and a combination on days 25 and 32 pi. The untreated control group received a placebo. In single-treatment groups, the efficacy of each treatment, measured by rate of reduction in muscle larvae, was significant (P0.05). We conclude that the efficacy of maslinic acid against larval stages of T. zimbabwensis in rats was comparable to that of fenbendazole, with no side-effects observed, making maslinic acid a promising anthelmintic against larval stages of Trichinella species.

A 38-year study on Trichinella spp. in wild boar (Sus scrofa) of Latvia shows a stable incidence with an increased parasite biomass in the last decade.

BACKGROUND: Trichinella spp. are zoonotic parasites transmitted to humans by the consumption of raw or insufficiently cooked meat of different animal species. The most common source of infection for humans is meat from pigs and wild boar (Sus scrofa). The aim of the present work was to evaluate the incidence of Trichinella spp. infections in wild boar hunted in Latvia over a 38 year interval (1976 to 2013). METHODS: A total 120,609 wild boars were individually tested for Trichinella spp. by trichinoscopy and, in case of negativity, by artificial digestion of 25 g muscles, in the 1976-2005 period, and by artificial digestion of 25-50 g muscles in the 2006-2013 period. Trichinella spp. larvae were identified at the species level by multiplex PCR. RESULTS: In the study period, the overall prevalence of infected wild boar was 2.5%. Trichinella britovi was the predominant (90%) species. The incidence of Trichinella spp. infection in wild boar exhibited two different trends. From 1976 to 1987, the incidence of infected/hunted wild boar increased from 0.23% to 2.56%, then it decreased to 0.19 in 1994. Thereafter, the incidence fluctuated between 0.05% and 0.37%. A statistically significant (P < 0.05) correlation (r = 0.54; p = 0.0199) was found between the trend of Trichinella spp. incidence in hunted wild boar and the number of snow cover days from 1976 to 1993. From 1997 to 2013, the estimated wild boar population of Latvia increased by 4.9 times and the hunting bag by 9.7 times, with a stable incidence of Trichinella spp. in the population. It follows that the biomass of Trichinella spp. larvae and of T. britovi, in particular, increased. CONCLUSIONS: The incidence trends of Trichinella spp. in wild boar could be related to the role played by the snow in reducing the thermal shock and muscle putrefaction which increases the survival of the larvae in muscle tissues of carrion in the 1976-1993 period; and, in the 1997-2013 period, to the increased biomass of Trichinella spp. due to the increased carnivore populations, which are the main reservoirs of these parasites.

High prevalence of Trichinella pseudospiralis in Florida panthers (Puma concolor coryi).

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.

Trend analysis of Trichinella in a red fox population from a low endemic area using a validated artificial digestion and sequential sieving technique.

Freezing of fox carcasses to minimize professional hazard of infection with Echinococcus multilocularis is recommended in endemic areas, but this could influence the detection of Trichinella larvae in the same host species. A method based on artificial digestion of frozen fox muscle, combined with larva isolation by a sequential sieving method (SSM), was validated using naturally infected foxes from Latvia. The validated SSM was used to detect dead Trichinella muscle larvae (ML) in frozen muscle samples of 369 red foxes from the Netherlands, of which one fox was positive (0.067 larvae per gram). This result was compared with historical Trichinella findings in Dutch red foxes. Molecular analysis using 5S PCR showed that both T. britovi and T. nativa were present in the Latvian foxes, without mixed infections. Of 96 non-frozen T. britovi ML, 94% was successfully sequenced, whereas this was the case for only 8.3% of 72 frozen T. britovi ML. The single Trichinella sp. larva that was recovered from the positive Dutch fox did not yield PCR product, probably due to severe freeze-damage. In conclusion, the SSM presented in this study is a fast and effective method to detect dead Trichinella larvae in frozen meat. We showed that the Trichinella prevalence in Dutch red fox was 0.27% (95% CI 0.065-1.5%), in contrast to 3.9% in the same study area fifteen years ago. Moreover, this study demonstrated that the efficacy of 5S PCR for identification of Trichinella britovi single larvae from frozen meat is not more than 8.3%.

The vasculature of nurse cells infected with non-encapsulated Trichinella species.

The vasculature surrounding the nurse cells of encapsulated Trichinella spiralis has been described previously. It has been postulated the function of these vessels is to support the growth of the parasite. We describe here for the first time the vasculature surrounding the nurse cells of non-encapsulated T. pseudospiralis and T. papuae. Similar to the vasculature of uninfected muscle cells, the vessels surrounding non-encapsulated Trichinella nurse cells are dense and branched longitudinally along the long axis of the muscle cells; they also appear to be similar in diameter. The netting pattern of enlarged vessels found around T. spiralis (encapsulated) nurse cells is not present in non-encapsulated Trichinella infections. The vessels surrounding non-encapsulated Trichinella nurse cells seem to exist prior to parasite invasion of the muscle cell.

Freeze-tolerance of Trichinella muscle larvae in experimentally infected wild boars.

Freeze-tolerance of encapsulated Trichinella muscle larvae (ML) is mainly determined by Trichinella species, but is also influenced by host species, the age of the infection and the storage time and temperature of the infected meat. Moreover, the freeze-tolerance of the encapsulated species appears to be correlated to the development of thick capsule walls which increases with age. An extended infection period and the muscle composition in some hosts (e.g. herbivores) may provide freeze-avoiding matrices due to high carbohydrate contents. The present experiment compares freeze-tolerance of Trichinella spiralis and Trichinella britovi ML in wild boar meat 24 weeks post inoculation (wpi). Three groups of four wild boars were infected with 200, 2000 or 20,000 ML of T. britovi (ISS 1575), respectively. Additionally, three wild boars were inoculated with 20,000 ML of T. spiralis (ISS 004) and two animals served as negative controls. All wild boars were sacrificed 24 wpi. Muscle samples of 70 g were stored at -21°C for 19, 30 and 56 h, and for 1-8 weeks. Larvae were recovered by artificial digestion. Their mobilities were recorded using Saisam(®) image analysis software and their infectivities were evaluated using mouse bioassays. Samples frozen for 19, 30 and 56 h allowed recovery of mobile ML, but samples frozen for 1 or 2 weeks did not. Correspondingly, only T. spiralis and T. britovi larvae isolated from wild boar meat frozen for 19, 30 and 56 h established in mice. This study showed that freezing at -21°C for 1 week inactivated T. spiralis and T. britovi ML encapsulated in wild boar meat for 24 weeks.

Unique antigenic gene expression at different developmental stages of Trichinella pseudospiralis.

Parasite-induced and parasite-regulated larval capsule formation and host immunosuppression are two major characteristics that are unique in Trichinella spp. infections, but the molecule(s) and mechanism(s) that mediate these processes remain largely unknown. Trichinella pseudospiralis and Trichinella spiralis, are obviously different with respect to these two characteristics. A comparative study of these two species, in particular their antigen expression profiles at different developmental stages (the main molecules involved in the cross-talk or interaction between each parasite and its host), may help us better understand the parasite molecules and mechanisms involved. Here, we constructed cDNA libraries from T. pseudospiralis adults (Ad), newborn larvae (NBL) and muscle larvae (ML) mRNA and screened them with pig anti-T. pseudospiralis serum collected 26, 32 and 60 days post-infection (p.i.). The most abundant antigens were found to vary among life-cycle stages. Pyroglutamy peptidase 1-like and 6-phosphogluconolactonase-like genes predominated in the Ad stage and a serine protease (SS2-1-like gene) predominated in NBL similar to that observed in T. spiralis. Muscle larvae expressed proteasome activator complex subunit 3-like and 21 kDa excretory/secretory protein-like genes. This study indicated that parasites of two species may utilise different molecules and mechanisms for larvae capsule formation and host immunosuppression during their infections. Proteins of antigenic genes identified in this study may be also good candidates for diagnosis, treatment or vaccination for T. pseudospiralis infection, and also for the differential diagnosis of two species' infections.

Susceptibility of laboratory rodents to Trichinella papuae.

Members of the genus Trichinella are small nematodes that can infect a wide range of animal hosts. However, their infectivity varies depending on the parasite and host species combination. In this study, we examined the susceptibility of 4 species of laboratory rodents, i.e., mice, rats, hamsters, and gerbils to Trichinella papuae, an emerging non-encapsulated Trichinella species. Trichinella spiralis and Trichinella pseudospiralis were also included in this study for comparison. Fifteen animals of each rodent species were infected orally with 100 muscle larvae of each Trichinella species. Intestinal worm burden was determined at day 6 and 10 post-inoculation (PI). The numbers of muscle larvae were examined at day 45 PI. The reproductive capacity index (RCI) of the 3 Trichinella species in different rodent hosts was determined. By day 6 PI, 33.2-69.6% of the inoculated larvae of the 3 Trichinella species became adult worms in the small intestines of the host animals. However, in rats, more than 96% of adult worms of all 3 Trichinella species were expelled from the gut by day 10 PI. In gerbils, only 4.8-18.1% of adult worms were expelled by day 10 PI. In accordance with the intestinal worm burden and the persistence of adults, the RCI was the highest in gerbils with values of 241.5±41.0 for T. papuae, 432.6±48 for T. pseudospiralis, and 528.6±20.6 for T. spiralis. Hamsters ranked second and mice ranked third in susceptibility in terms of the RCI, Rats yielded the lowest parasite RCI for all 3 Trichinella species. Gerbils may be an alternative laboratory animal for isolation and maintenance of Trichinella spp.

Susceptibility of Laboratory Rodents to Trichinella papuae

Members of the genus Trichinella are small nematodes that can infect a wide range of animal hosts. However, their infectivity varies depending on the parasite and host species combination. In this study, we examined the susceptibility of 4 species of laboratory rodents, i.e., mice, rats, hamsters, and gerbils to Trichinella papuae, an emerging non-encapsulated Trichinella species. Trichinella spiralis and Trichinella pseudospiralis were also included in this study for comparison. Fifteen animals of each rodent species were infected orally with 100 muscle larvae of each Trichinella species. Intestinal worm burden was determined at day 6 and 10 post-inoculation (PI). The numbers of muscle larvae were examined at day 45 PI. The reproductive capacity index (RCI) of the 3 Trichinella species in different rodent hosts was determined. By day 6 PI, 33.2-69.6% of the inoculated larvae of the 3 Trichinella species became adult worms in the small intestines of the host animals. However, in rats, more than 96% of adult worms of all 3 Trichinella species were expelled from the gut by day 10 PI. In gerbils, only 4.8-18.1% of adult worms were expelled by day 10 PI. In accordance with the intestinal worm burden and the persistence of adults, the RCI was the highest in gerbils with values of 241.5+/-41.0 for T. papuae, 432.6+/-48 for T. pseudospiralis, and 528.6+/-20.6 for T. spiralis. Hamsters ranked second and mice ranked third in susceptibility in terms of the RCI, Rats yielded the lowest parasite RCI for all 3 Trichinella species. Gerbils may be an alternative laboratory animal for isolation and maintenance of Trichinella spp.

The different infectivity of Trichinella spiralis and Trichinella nativa in rat does not solely localize to enteral or parenteral phase.

Rats are selective hosts for Trichinella nativa; infection levels in their muscles remain low when compared to Trichinella spiralis. To identify in which phase of the life cycle the selective responses take place, rats were infected perorally (p.o.) and intravenously (i.v.) with T. spiralis and T. nativa. Six rats were placed in each group; three rats served as uninfected controls. The infection intensities were examined 5 or 6 weeks after the infection. T. spiralis muscle larva burden was 57 times higher compared to that of T. nativa after p.o. inoculation (mean larvae per gram ± SD, 1,243 ± 378.9 vs. 22 ± 21.5). In i.v. inoculation, 12 times more T. spiralis larvae reached the muscle than with T. nativa. Also, in vitro newborn larva (NBL) production at day 6 postinfection was analyzed for parasites originating from both rat and mouse. The mean number of in vitro-produced NBL of rat-origin T. spiralis during 24 h was 70 (95 % CI, 65-75) and of T. nativa, 23 (95 % CI, 21-24). Intriguingly, this difference was less obvious with mouse-origin female Trichinella. NBL production of mouse origin T. spiralis was 57 (95 % CI, 51-62) and T. nativa, 29 (95 % CI, 26-31). Intestine histology at day 40 post p.o. infection, serological response, and weight dynamics were indistinguishable between the Trichinella species but differed from controls. Our results show that the defense against T. nativa is not solely enteral in rats but may partly be explained by different reproduction processes for the two Trichinella species in rat.

Immunomodulation in trichinellosis: does Trichinella really escape the host immune system?

This review describes different aspects of the host immune response to Trichinella. The role of antibodies, T cells, mast cells, eosinophils and neutrophils in immune reaction to this nematode is considered, in the light of the recent data derived from experimental models, both in in vivo and in vitro. The knowledge of immune response mechanisms against Trichinella is fundamental to understand how the parasite can escape such mechanisms. The principal evasion mechanisms of host immune response occurring in trichinellosis are described, some of which are shared by other parasites, some others are peculiar of this parasite, but particular attention is focused on immunomodulation and the possibilities to exploit this parasite ability to verify the effects on immuno-mediated diseases. In conclusion, some considerations on the actual ability to escape the host immune response by the parasite are discussed, taking into account the recent data that shows that the parasite might rather drive immune system of the host towards a less dangerous response.