Effect of temperature on the survival of Alaria alata mesocercariae.

Recent findings of Alaria alata mesocercariae in wild boars and other animals in Europe reinforced the concern about the public health risk posed by this parasite especially if the game meat is insufficiently heated during preparation. Cooking and freezing are effective methods for the inactivation of parasites in meat whereas refrigeration is considered as an essential part of the Good Hygiene Practice. Additionally, microwave dielectric heating may represent an equally effective tool for parasite inactivation. Therefore, isolated vital mesocercariae were examined with respect to their resilience against heating, refrigeration, freezing, and microwave heating. A. alata mesocercariae stored in Ringer's solution do not survive heating temperatures that exceed 60.0 °C. Similarly, exposure to microwave heating ensured an inactivation of all parasite developmental stages after 90 s of treatment. In contrast, the parasites' tolerance towards cold is far higher as the mesocercariae survived refrigeration temperatures (4.0 ± 2 °C) in Ringer's solution for up to 13 days. An effective inactivation by cold is therefore only guaranteed if the infested game meat is frozen to a core temperature of -13.7 °C for a minimum of 2 h at least. Game meat should be handled with the same or even higher caution than meat of husbandry animals since wild animals may be infected with parasites or other zoonotic agents that are not common in livestock. It is therefore of crucial importance that appropriate temperature time protocols are used for the reliable inactivation of these zoonotic agents.

Effects of in vitro conditions on the survival of Alaria alata mesocercariae.

The aim of this study was to determine the effect of different concentrations of table salt (NaCl) and ethanol (v/v) solutions on the viability of Alaria alata mesocercariae. Furthermore, the survival of A. alata mesocercariae during simulated human gastric digestion was evaluated. For this purpose, A. alata mesocercariae migration technique (AMT) was used for the isolation of the parasite from high-positive A. alata mesocercariae meat from wild boar, raccoon, raccoon dog, and badger meat. In total, we have studied the behavior of 582 larvae under different conditions (NaCl, ethanol, and artificial gastric juice) in three independent in vitro experiments. The larvae survived at a NaCl concentration of up to 2.0% until day 21 with a median survival time of 11 days. At 3.0% NaCl concentration, the larvae lost their vitality after less than 24 h. In addition, it was found that ethanol concentrations from 8.0 to 70.0% were effective at reducing survival of A. alata mesocercariae within a short period of time (<1 min). Finally, our studies have revealed that it required 120 min to reliably inactivate all A. alata mesocercariae within HCl-pepsin digestion solution with a pH of 1.5-2.0 at 37°C. Consequently, the results showed that 3.0% is the minimum concentration of NaCl in meat products recommended for human consumption because at lower NaCl concentration the parasite survived for a substantial period of time. Finally, the common concentrations of ethanol used for the disinfection of surfaces in household and/or laboratory, are sufficient for the inactivation of A. alata mesocercariae.

Tenacity of Alaria alata mesocercariae in homemade German meat products.

A renewed interest in the pathogenic potential of Alaria alata mesocercariae emerged over the last 10years as a result of increased findings of this parasite in feral pigs during official examination for Trichinella spp. Cases of food associated human alariosis in North America suggest that a risk associated with the consumption of traditional raw cured products from infected wild boar meat cannot be neglected because the commonly applied preservation techniques may not necessarily kill the mesocercariae. In addition, changes in consumer behavior and new preparation methods for game meat (e.g. pink roasting and grilling) may increase the risk for food-associated parasitic infections. Thus, there is a strong need for the evaluation of the tenacity of A. alata mesocercariae against different physical and chemical influences as pertaining to common preservation and preparation techniques. Against this backdrop the aim of our work was a sound analysis of the survivability of A. alata mesocercariae during curing, fermentation, cold smoking and drying in raw cured meat products. Eighty three samples of traditional German meat products were prepared from naturally infected game meat and partly spiked with additional vital mesocercariae to achieve an adequate dose of infection. The resultant products were examined chronologically for dead and viable A. alata mesocercariae with the Alaria mesocercariae migration technique. After 24h of production, vital A. alata mesocercariae were still found in raw type sausages but no vital parasites were detected in the final products. Based on these results a possible risk for the consumer for an infection with A. alata mesocercariae through the consumption of contaminated raw cured products can be largely ruled out if the respective food technological procedures are carried out properly. However, a risk for the consumer cannot be excluded in cases of very early consumption of these products.

In-house validation of an improved sample extraction and clean-up method for GC determination of isomers of nervonic acid in meat products.

An improved extraction and clean-up method for determination of brain-specific fatty acids, in particular lignoceric acid (C24:0) and the cis/ trans isomers of nervonic acid (15 c-t C24:1), in meat products has been developed. The method is based on isolation of the polar lipids of interest from the bulk lipids by solid-phase extraction. The fatty acids, derivatised to their fatty acid methyl esters, are quantified by GC in a DB5 column. Fresh meat samples were extracted by using a mixture of n-butanol:hexane (1:9) as solvent. The extract was loaded in a silica gel cartridge column previously equilibrated with hexane. The first fraction containing the major part of the fat was eluted with hexane while acetone and methanol allowed the elution of fatty acids bound to polar moieties such as nervonic and lignoceric acids. This second fraction containing the analyte was methylated and injected into the GC for quantification after addition octacosane (C(28)) as internal standard.