Topics for training activities

Anisakis

The larval stage of nematodes of the Anisakidae family parasitize fishes, cephalopods and shrimps, whereas the adult worm parasitize fishes, marine birds and mammals. In humans, ingestion of the larvae of the genus Anisakis and Pseudoterranova cause the diseases known as anisakiasis and anisakidosis, respectively. Anisakis and Pseudoterranova can be distinguished morphologically only at the genus level. For species identification, the EURLP has developed and accredited the following molecular methods:

• identification at species level of parasites of the family Anisakidae by PCR/RFLP and multiplex PCR

Cryptosporidium

Parasites of the genus Cryptosporidium cause enteritis in humans and animals, and can be transmitted by multiple routes. Numerous outbreaks linked to contaminated food and water occur yearly in Europe. The EURLP has long-lasting expertise in the detection of the parasite in clinical and environmental samples, in molecular typing and epidemiology, and in comparative genomics. The accredited method developed by EURLP is:

• identification at species level of Cryptosporidium oocysts by PCR/RFLP

Echinococcus

Echinococcus granulosus sensu lato and Echinococcus multilocularis are zoonotic parasites causing cystic and alveolar echinococcosis, respectively. These cestode infections are of global health relevance and ranked as the most important food-borne parasitic diseases at European level. They infect a wide spectrum of definitive and intermediate animal hosts, causing relevant morbidity and mortality in humans. The EURLP has long-lasting expertise in the molecular and clinical epidemiology and detection of these parasites. The accredited method developed by the EURLP is:

• identification of Echinococcus granulosus sensu lato complex at species/genotypes level by PCR/RFLP and multiplex PCR

Giardia

The flagellate Giardia duodenalis causes enteritis in humans and animals, and possess multiple transmission routes. Outbreaks linked to contaminated food and water are often reported across Europe. The EURLP has long-lasting expertise in the detection of the parasite in clinical and environmental samples, in molecular typing and epidemiology, and in cellular biology. The EURLP has developed and accredited the following methods:

• identification of the assemblages A and B of Giardia duodenalis by PCR

• identification at assemblage level of Giardia duodenalis cysts by PCR/RFLP

Opisthorchis

Opisthorchis felineus and Opisthorchis viverrini are foodborne zoonotic parasites found in Europe and Asia, respectively. Humans can get the infection by eating raw or undercooked freshwater fish infected with the larval stage metacercariae. Diagnosis of opisthorchiasis is based on identification of parasite eggs in stool specimens and on serological tests. The EURLP has developed and accredited the following diagnostic methods:

• identification of Opisthorchis spp eggs by PCR

• detection of anti-Opisthorchis antibodies in human sera

Toxoplasma

Toxoplasma gondii is a protozoan parasite able to infect all warm-blooded animals (mammals and birds), including humans. Humans can acquire toxoplasmosis by the ingestion of fruits, vegetables or water contaminated by oocysts, or by the consumption of raw or undercooked meat containing tissue cysts. Sensitive detection of T. gondii in foodstuff is best achieved using molecular methods. For this purpose, the EURLP has developed and accredited the following method:

• identification of Toxoplasma gondii DNA in food matrices (fresh/processed meat and leafy green vegetable) by LAMP

Trichinella

The Trichinella genus comprises zoonotic parasites found worldwide in many carnivorous and omnivorous warm-blooded and cold-blooded animals. The disease in humans, i.e., trichinellosis, is caused by the ingestion of raw or undercooked meat containing larvae of Trichinella species. The EURLP has long-lasting expertise in the detection and epidemiology of the parasite, in molecular identification at species level, and in the organization of proficiency testing. As OIE Reference Laboratory and International Trichinella Reference Center, the EURLP has developed and accredited the following diagnostic methods:

• artificial digestion to detect Trichinella larvae in meat samples according to ISO 18743:2015

• identification of Trichinella muscle larvae at species level by Multiplex PCR

• detection of anti-Trichinella IgG in serum from infected host by indirect ELISA

• identification of Trichinella spp. proteins recognized by specific IgG in serum of infected hosts by western blotting

The EURLP can also offer support in the following topics:

• maintenance of Trichinella isolates in vivo

• production of ES antigen from Trichinella muscle larvae in culture

• organization of proficiency testing on the detection and/or species identification of Anisakis, Echinococcus, Toxoplasma and Trichinella