[Assessment of transmammary transmission of Trichinella patagoniensis in BALB/c mice]. / Evaluación de la transmisión galactógena de Trichinella patagoniensis en ratones BALB/c.

Since 1916 to date, it has been suspected that vertical transmission of parasites from the genus Trichinella could occur in pregnant or lactating women during the parenteral phase of infection. The aim of the present study was to evaluate the transmammary transmission of T. patagoniensis in BALB/c mice. Twenty 7-week-old BALB/c mice were distributed into two groups of 10 individuals each, depending on the time of gestation when they were infected, 15 or 18 days after detection of the vaginal plug. Each group was subdivided into two subgroups of 5 mice each, which were given an oral dose of 100 or 500 infective larvae respectively. Euthanasia and subsequent artificial digestion was performed in the pups and the dams. No T. patagoniensis L1 larvae were found in any of the offsprings analyzed. The observed results suggest that vertical transmission of T. patagoniensis would not be possible in BALB/c mice.

First report of Toxocara cati in the domestic land snail Rumina decollata.

The prospective role of the land snail Rumina decollata as a potential paratenic host of Toxocara cati for domestic cats was studied. R. decollata specimens and cats' feces were collected from the open spaces of a Buenos Aires city hospital. Cats' feces were analyzed and snails were digested to identify T. cati stages, by morphological and molecular analyses. T. cati larval eggs were recovered from 23.5% (4/17) of the sampled feces. Twenty percent of snail pools (5/25) were confirmed to be positive for Toxocara spp. third larval stage (L3) by PCR. The mean value of total larvae recovered per gram of snail in all positive pools was 5.1, with a maximum 33 L3/pool. This is the first report of T. cati infective larvae in R. decollata domestic snail as a paratenic host, since the relationship between infection in snails and in cats' feces could be demonstrated in a common environment.

Reprint of: 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.

Reprint of: Gamma radiation effect on Trichinella pseudospiralis and Trichinella spiralis infected wild boar meat.

Trichinellosis is a foodborne parasitic disease caused by Trichinella spp. Different methods, such as cooking, freezing and irradiation, have been suggested to inactivate the parasite in meat infected with Trichinella spp. The International Commission on Trichinellosis (ICT) recommends an irradiation dose of 0.3 kGy to inactivate Trichinella spp. larvae, but its effectiveness in all Trichinella species has not yet been demonstrated. Therefore, the objective of the present study was to determine the effectiveness of gamma radiation in treating wild boar meat to inactivate Trichinella pseudospiralis and Trichinella spiralis. Two wild boars (Sus scrofa) were each inoculated per os (PO) with 20,000 muscle larvae (ML) of Trichinella. One wild boar was inoculated with T. pseudospiralis and the other one with T. spiralis. Both wild boars were euthanized 20 weeks post infection (pi). A 250░g sample from each selected muscle type (side ribs, shoulder and upper forelegs) was obtained in duplicate from each animal. One group of samples was not irradiated (Gc; n░=░6) while the other group of samples was irradiated (Gt; n░=░6) at the Atomic Centre of the Argentinean National Commission of Atomic Energy (CNEA) with a minimum and maximum dose of 0.32-0.41 kGy. Twenty gram of each muscle from Gt and Gc were taken at 24░h, 7 days, 14 days and 21 days post-irradiation, and all samples were individually processed by means of artificial digestion. The ML were then inoculated into mice to evaluate the intestinal phase and the muscular phase. All recovered larvae from Gt and Gc samples showed integrity of the cuticle and active motility. Adult worms and ML were recovered from all mice inoculated with ML from Gc. However, no adult worms or ML were obtained in mice inoculated with ML from Gt. These results show the efficacy of irradiation to inhibit the development of T. pseudospiralis and T. spiralis in the host, which could possibly be extended to other non-encapsulated species, but further studies are needed to demonstrate this hypothesis.

Reprint of: A review on Trichinella infection in South America.

Trichinella spp. causes human trichinellosis by means of the consumption of raw or inadequately treated meat from domestic or game animals. In the Americas, as well as in other continents, Trichinella infection is a health issue for humans and has a negative impact on the pork meat market, generated by people's fear of becoming infected with the parasite. The distribution of human cases and the sources of this disease in humans and animals were analysed in this report, which summarizes the information available regarding Trichinella infection in animals and humans in South America. Within South America, human infection with Trichinella was documented in Argentina and Chile during the period 2005-2019. Trichinellosis is endemic in these countries for, with human cases and foci in domestic and wild animals. In Argentina, human cases occur throughout the country, with foci found in pigs and wild animals. In Argentina, during the period 2012-2018, the number of suspected human cases reached 6,662. T. spiralis was identified in one South American sea lion (Otaria flavescens) from Patagonia, Argentina, for the first time in the region in 2018. In Chile, 258 human cases of trichinellosis were confirmed during the period 2005-2015; out of those 258 cases, most samples which tested positive for Trichinella spp. (29.5 %) were detected in the Metropolitan district (Santiago de Chile and outskirts), and 17.4 % in The Lake district. Regarding age brackets, people between 30-49 years of age showed the most cases (40.1 %). In Brazil, the infection is absent in domestic species but it has been found in wild boars (Sus scrofa) but limited to one or more region of the country. Within the animal species destined for food in South America, those that showed higher parasitical loads were pigs and wild boars, while armadillos (Chaetophractus villosus) and peccaries (Tayassu tajacu) showed very low Trichinella spp. larvae loads (0.04-0.1 larvae/g). Antibodies against Trichinella spp. have been detected in pigs from Ecuador and Bolivia. In Bolivia, antibodies were also found in humans. Peru, Colombia and Uruguay have no documented presence of Trichinella spp. in animals and humans. There is insufficient information regarding the presence of Trichinella spp. in domestic and wild animals, as well as in humans, since only a very limited number of surveys have been carried out. No papers with information on Trichinella spp. circulating in animals or humans have been published regarding the situation in Guyana, Surinam, French Guiana, Venezuela and Paraguay. Considering the growth of the guinea pig meat market in the Andean region, and the high prevalence of the disease reported in free range pigs and wild boars, as well as other game animal species, it is important to focus on the role of biosecurity and risk management, while improving meat market regulations, and detection of infection prior to consumption, in order to reduce the risk of transmission of this zoonotic disease to humans.

Trichinella infection in wild animals from endemic regions of Argentina.

Natural infection with Trichinella has been described in more than 150 mammalian species. However, few reports of Trichinella infection in wild animals have come from Argentina. In this study, muscle tissue was obtained from wild animals in Argentina with the aim of evaluating the presence of Trichinella. A total of 169 muscle samples were collected to determine the presence of Trichinella larvae by artificial digestion. The 169 muscle samples originated from 12 species including 36 opossums (Didelphis albiventris), 19 armadillos (Chaetophractus villosus), 9 capybaras (Hydrocaeris hydrocaeris), 1 puma (Puma concolor), 3 grey fox (Lycalopex gymnocercus), 6 coypus (Myocastor coypus), 6 skunks (Conepatus chinga), 2 ferrets (Galictis cuja), 66 rats (Rattus norvegicus), 6 mice (Mus musculus), 12 wild boars (Sus scrofa), and 3 wild cats (Felis geoffroyi). Trichinella infection was detected in 1 puma [2 larvae per gram (LPG)], 3 wild boars (8-420 LPG), 3 armadillos (0.04-0.08 LPG), and 9 rats (0.1 to 150 LPG). Only 3 Trichinella isolates, of 1 rat and 2 wild boars from Neuquén, were identified as Trichinella spiralis by nested PCR. The presence of Trichinella infection among wild animal populations suggests a sylvatic cycle of transmission in Argentina, which can serve as a reservoir for humans and domestic animals. Further, evidence of high prevalence in rats emphasizes the need to improve pig management, mainly in small individual farms without adequate technology, to enhance the quality of feeds, and to improve veterinary services to avoid exposure of pigs to Trichinella.

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.

Gamma radiation effect on Trichinella pseudospiralis and Trichinella spiralis infected wild boar meat.

Trichinellosis is a foodborne parasitic disease caused by Trichinella spp. Different methods, such as cooking, freezing and irradiation, have been suggested to inactivate the parasite in meat infected with Trichinella spp. The International Commission on Trichinellosis (ICT) recommends an irradiation dose of 0.3 kGy to inactivate Trichinella spp. larvae, but its effectiveness in all Trichinella species has not yet been demonstrated. Therefore, the objective of the present study was to determine the effectiveness of gamma radiation in treating wild boar meat to inactivate Trichinella pseudospiralis and Trichinella spiralis. Two wild boars (Sus scrofa) were each inoculated per os (PO) with 20,000 muscle larvae (ML) of Trichinella. One wild boar was inoculated with T. pseudospiralis and the other one with T. spiralis. Both wild boars were euthanized 20 weeks post infection (pi). A 250 g sample from each selected muscle type (side ribs, shoulder and upper forelegs) was obtained in duplicate from each animal. One group of samples was not irradiated (Gc; n = 6) while the other group of samples was irradiated (Gt; n = 6) at the Atomic Centre of the Argentinean National Commission of Atomic Energy (CNEA) with a minimum and maximum dose of 0.32 - 0.41 kGy. Twenty gram of each muscle from Gt and Gc were taken at 24 h, 7 days, 14 days and 21 days post-irradiation, and all samples were individually processed by means of artificial digestion. The ML were then inoculated into mice to evaluate the intestinal phase and the muscular phase. All recovered larvae from Gt and Gc samples showed integrity of the cuticle and active motility. Adult worms and ML were recovered from all mice inoculated with ML from Gc. However, no adult worms or ML were obtained in mice inoculated with ML from Gt. These results show the efficacy of irradiation to inhibit the development of T. pseudospiralis and T. spiralis in the host, which could possibly be extended to other non-encapsulated species, but further studies are needed to demonstrate this hypothesis.

A review on Trichinella infection in South America.

Trichinella spp. causes human trichinellosis by means of the consumption of raw or inadequately treated meat from domestic or game animals. In the Americas, as well as in other continents, Trichinella infection is a health issue for humans and has a negative impact on the pork meat market, generated by people's fear of becoming infected with the parasite. The distribution of human cases and the sources of this disease in humans and animals were analysed in this report, which summarizes the information available regarding Trichinella infection in animals and humans in South America. Within South America, human infection with Trichinella was documented in Argentina and Chile during the period 2005-2019. Trichinellosis is endemic in these countries for, with human cases and foci in domestic and wild animals. In Argentina, human cases occur throughout the country, with foci found in pigs and wild animals. In Argentina, during the period 2012-2018, the number of suspected human cases reached 6,662. T. spiralis was identified in one South American sea lion (Otaria flavescens) from Patagonia, Argentina, for the first time in the region in 2018. In Chile, 258 human cases of trichinellosis were confirmed during the period 2005-2015; out of those 258 cases, most samples which tested positive for Trichinella spp. (29.5%) were detected in the Metropolitan district (Santiago de Chile and outskirts), and 17.4% in The Lake district. Regarding age brackets, people between 30-49 years of age showed the most cases (40.1%). In Brazil, the infection is absent in domestic species but it has been found in wild boars (Sus scrofa) but limited to one or more region of the country. Within the animal species destined for food in South America, those that showed higher parasitical loads were pigs and wild boars, while armadillos (Chaetophractus villosus) and peccaries (Tayassu tajacu) showed very low Trichinella spp. larvae loads (0.04 - 0.1 larvae/g). Antibodies against Trichinella spp. have been detected in pigs from Ecuador and Bolivia. In Bolivia, antibodies were also found in humans. Peru, Colombia and Uruguay have no documented presence of Trichinella spp. in animals and humans. There is insufficient information regarding the presence of Trichinella spp. in domestic and wild animals, as well as in humans, since only a very limited number of surveys have been carried out. No papers with information on Trichinella spp. circulating in animals or humans have been published regarding the situation in Guyana, Surinam, French Guiana, Venezuela and Paraguay. Considering the growth of the guinea pig meat market in the Andean region, and the high prevalence of the disease reported in free range pigs and wild boars, as well as other game animal species, it is important to focus on the role of biosecurity and risk management, while improving meat market regulations, and detection of infection prior to consumption, in order to reduce the risk of transmission of this zoonotic disease to humans.

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.

Larval development of Aelurostrongylus abstrusus in experimentally infected Rumina decollata snails.

Aelurostrongylus abstrusus is a lungworm distributed worldwide that affects wild and domestic cats, causing bronchopneumonia of varying intensity. Snails serve as intermediate hosts. The aim of the present study was to assess the larval development of A. abstrusus in R. decollata snails and to investigate its potential as an intermediate host. For this purpose, first-stage larvae (L1) of A. abstrusus were obtained from the faeces of naturally infected cats. Doses of 500 L1/snail were given to 24 R. decollata snails, placed on the soil of the breeder chamber, and maintained under laboratory conditions. Three snails were killed at 8, 10, 12, 16, 22, 26, 45 and 55 days post-infection (dpi), and the muscular foot and visceral body were separately digested by an artificial digestion technique. The morphometric parameters of different larval stages were recorded. The mean number of larvae reaching the infective stage at the end of the study (L3) was 262 larvae/snail. The greatest development to L3 was recorded from days16 to 55 pi, during which the isolation was maximum. A. Abstrusus L3 were isolated from the viscera, but isolation from the snail foot was significantly higher. Our results showed for the first time the ability of A. Abstrusus larvae to develop in R. decollata, thus serving as a potential intermediate host.

Primer reporte de Toxocara cati en el caracol doméstico Rumina decollata / First report of Toxocara cati in the domestic land snail Rumina decollata

Se estudió el rol prospectivo de Rumina decollata como potencial hospedador paraténico de Toxocara cati para los gatos domésticos. Se recolectaron caracoles R. decollata y heces de gatos de un hospital de la Ciudad Autónoma de Buenos Aires. Se procesaron las heces y los caracoles fueron digeridos para identificar estadios de T. cati por análisis morfológico y molecular. El 23,5% (4/17) de las muestras de heces resultaron positivas a huevos larvados de T. cati. El 20% (5/25) de los pooles de caracoles fue positivo a larvas de tercer estadío (L3) de Toxocara spp. por PCR. El promedio de larvas totales recuperadas por gramo de caracol en todos los pooles positivos fue de 5.1, con un máximo de 33 L3/pool. Se trata del primer reporte de R. decollata como hospedador paraténico de T. cati, puesto que ha sido demostrada la infección en caracoles y gatos en un ambiente común

The prospective role of the land snail Rumina decollata as a potential paratenic host of Toxocara cati for domestic cats was studied. R. decollata specimens and cats' feces were collected from the open spaces of a Buenos Aires city hospital. Cats' feces were analyzed and snails were digested to identify T. cati stages, by morphological and molecular analyses. T. cati larval eggs were recovered from 23.5% (4/17) of the sampled feces. Twenty percent of snail pools (5/25) were confirmed to be positive for Toxocara spp. third larval stage (L3) by PCR. The mean value of total larvae recovered per gram of snail in all positive pools was 5.1, with a maximum 33 L3/pool. This is the first report of T. cati infective larvae in R. decollata domestic snail as a paratenic host, since the relationship between infection in snails and in cats' feces could be demonstrated in a common environment