Effect of co-encapsulation using a calcium alginate matrix and fructooligosaccharides with gelatin coating on the survival of Lactobacillus paracasei cells.

The demand for functional foods is increasing each year because consumers are gaining awareness about the importance of a healthy diet in the proper functioning of the body. Probiotics are among the most commonly known, commercialized, and studied foods. However, the loss of viability of probiotic products is observed during their formulation, processing, and storage. This study aimed to investigate the co-encapsulation of two Lactobacillus paracasei probiotic strains (LBC81 and ELBAL) with fructooligosaccharides (FOS) in a calcium alginate matrix using extrusion technology with gelatin as a coating material. The viability of the strains under gastrointestinal conditions and in storage at low temperature was also assessed. An immobilization yield of more than 59% was observed for both bacterial strains. Exposure to 2% biliary salts led to a decrease in the viability of free cells in the two L. paracasei strains, whereas the viability of microencapsulated cells increased up to 47%. After 35 days of storage at 4°C, the population of free cells was reduced, but microencapsulated cells remained stable after storage at low temperature. LBC81 bacteria microencapsulated with 1.5% FOS coated with gelatin were the most resistant to the stressful environments tested. Therefore, these results showed that co-encapsulation with FOS in a calcium alginate matrix coated with gelatin improved L. paracasei survival and may be useful for the development of more resistant probiotics and new functional foods.

Differential secretome profiling of a swine tracheal cell line infected with mycoplasmas of the swine respiratory tract.

Mycoplasma hyopneumoniae and Mycoplasma flocculare are genetically similar. However, M. hyopneumoniae causes porcine enzootic pneumonia, while M. flocculare is a commensal bacterium. M. hyopneumoniae and M. flocculare do not penetrate their host cells, and secreted proteins are important for bacterium-host interplay. Thus, the secretomes of a swine trachea cell line (NPTr) infected with M. hyopneumoniae 7448 (a pathogenic strain), M. hyopneumoniae J (a non-pathogenic strain) and M. flocculare were compared to shed light in bacterium-host interactions. Medium from the cultures was collected, and secreted proteins were identified by a LC-MS/MS. Overall numbers of identified host and bacterial proteins were, respectively, 488 and 58, for NPTr/M. hyopneumoniae 7448; 371 and 67, for NPTr/M. hyopneumoniae J; and 203 and 81, for NPTr/M. flocculare. The swine cells revealed different secretion profiles in response to the infection with each M. hyopneumoniae strain or with M. flocculare. DAMPs and extracellular proteasome proteins, secreted in response to cell injury and death, were secreted by NPTr cells infected with M. hyopneumoniae 7448. All three mycoplasmas secreted virulence factors during NPTr infection, but M. hyopneumoniae 7448 secreted higher number of adhesins and hypothetical proteins, that may be related with pathogenicity. SIGNIFICANCE: The enzootic pneumonia caused by mycoplasmas of swine respiratory tract has economic loss consequences in pig industry due to antibiotic costs and pig weight loss. However, some genetically similar mycoplasmas are pathogenic while others, such as Mycoplasma hyopneumoniae and Mycoplasma flocculare, are non-pathogenic. Here, we conducted an infection assay between swine cells and pathogenic and non-pathogenic mycoplasmas to decipher secreted proteins during host-pathogen interaction. Mycoplasma response to cell infection was also observed. Our study provided new insights on secretion profile of swine cells in response to the infection with pathogenic and non-pathogenic mycoplasmas. It was possible to observe that pathogenic M. hyopneumoniae 7448 secreted known virulence factors and swine cells responded by inducing cell death. Otherwise, M. hyopneumoniae J and M. flocculare, non-pathogenic mycoplasmas, secreted a different profile of virulence factors in response to swine cells. Consequently, swine cells altered their secretome profile, but the changes were not sufficient to cause disease.