A new molecular approach to assess the occurrence of Sarcocystis spp. in cattle and products thereof: preliminary data

The genus Sarcocystis consists of more than 200 species. Those protozoa are characterised by a biological cycle composed by two obligatory hosts, definitive and intermediate. Apart from being possibly pathogenic for the intermediate host, a number of authors consider the intestinal sarcocystosis a minor zoonotic disease. Humans, in fact, can act as definitive host for two sarcosporidian species, S. suihominis e S. hominis, being infected through the consumption of raw or undercooked pig and bovine meat, respectively. Other two species could parasitise cattle: S. cruzi and S. hirsuta, having canids and felids as definitive hosts, respectively. The three species differentiate from each other in dimensions and cystic wall morphology, this latter being the basis for taxonomical studies. In 2010, the European Food Safety Authority (EFSA) highlighted the absence of reliable methods for epidemiological studies on the presence of Sarcocystis spp. in animals and products thereof. On this basis, the present study has been developed a new molecular method for the identification of Sarcocystis in bovine meat. For the development of the polymerase chain reaction (PCR) protocol, a set of samples of bovine meat from cattle (N=15), slaughtered at the didactic abattoir at the Veterinary Faculty of Turin University, has been collected, sequenced and used as reference samples during the study. A second set of samples (N=29), gathered from the same abattoir (N=12) and from abattoirs of Piedmont region (N=17), has been used for applicability tests. The overall positive rate for Sarcocystis spp. in our samples has been 91% (40/44), with S. cruzi representing the species with higher rates (68%), followed by S. hominis (43%) and S. hirsuta (2%). Based on the results of specificity and applicability tests performed in this study, the newly developed protocol proved to be reliable and suitable for epidemiologic purposes

Mycobiota composition of robiola di roccaverano cheese along the production chain

Robiola di Roccaverano is a Protected Designation of Origin (PDO) cheese from the Piedmont region of Italy. In this study, the mycobiota occurring during Robiola di Roccaverano production was elucidated. Samples of milk, Natural Milk Cultures (NMC), curd, 5- and 15-days ripened cheese were collected from one dairy plant and the mycobiota was analyzed by the metataxonomic approach. Milk samples showed a high diversity and Cladosporium, Kluyveromyces marxianus, Geotrichum candidum and Debaryomyces hansenii were found with higher relative abundance. This mycobiota remains quite stable in NMC and curd matrices although the relative abundance of K. marxianus and G. candidum yeasts increased significantly and shaped the fungal composition of 5- and 15-day ripened cheese

Distribution of Pseudomonas species in a dairy plant affected by occasional blue discoloration

During 2010 many cases of discoloration in mozzarella, popularly termed as blue mozzarella, have been reported to the attention of public opinion. Causes of the alteration were bacteria belonging to the genus Pseudomonas. The strong media impact of such cases has created confusion, not only among consumers, but also among experts. In order to help improving the knowledge on microbial ecology of this microorganism a study has been set up with the collaboration of a medium-sized dairy plant producing fresh mozzarella cheese, with occasional blue discoloration, conducting surveys and sampling in the pre-operational, operational and post-operational process phase, milk before and after pasteurization, water (n=12), environmental surfaces (n=22) and the air (n=27). A shelf life test was conducted on finished products stored at different temperatures (4-8°C). Among the isolates obtained from the microbiological analysis of the samples, 60 were subjected to biomolecular tests in order to confirm the belonging to Pseudomonas genus and to get an identification at species level by the amplification and sequencing of the gyrB gene. The results of microbiological tests demonstrated the presence of microorganisms belonging to the genus Pseudomonas along the entire production lane; molecular tests showed 7 different species among the 40 isolates identified. One particular species (Pseudomonas koreensis) was isolated from blue discolored mozzarella cheese and was indicated as the most relevant for the production plant, both for the distribution along the processing chain and for the consequences on the finished product