Salmonella detection with LAMP and qPCR and identification of serovars of interest by multiplex qPCR in poultry carcasses.

Salmonella is present in the poultry production chain and is a major challenge in terms of food safety and animal health. The early Salmonella detection is one of the main tools to control and prevent the transmission of this pathogen. Microbiological isolation and serotyping to identify and differentiate Salmonella serovars are laborious processes, time-consuming, and expensive. Therefore, molecular diagnostic methods can be rapid and efficient alternatives to the detection of this pathogen. Thus, the aim herein was to standardize and evaluate the use of loop-mediated isothermal amplification (LAMP) in comparison with real-time PCR (qPCR) for detection of Salmonella associated with a multiplex qPCR for simultaneous identification and differentiation of S. Enteritidis, S. Typhimurium, S. Pullorum, and S. Gallinarum. The LAMP, qPCR, and multiplex qPCR assays were comparable in specificity. The three techniques were evaluated for specificity for 16 different serovars of Salmonella and for 37 strains of the serovars of interest. The limit of detection and the efficiency of the LAMP, qPCR, and multiplex qPCR reactions were determined. The techniques were applied to 33 samples of chicken carcasses and compared to the results of conventional microbiology for validation. As results, LAMP was specific in the detection of different Salmonella serovars but presented lower limit of detection ranging from 101 to 104 CFU/reaction. In comparison, qPCR could detect less cells (100 to 102 CFU/reaction), reaching equal specificity and better repeatability in the assays. The qPCR multiplexing for identification of the different serovars also showed good specificity, with the detection threshold between entre 101 and 102 CFU/reaction. The results obtained in the analyses on poultry carcasses suggested a correspondence between the results obtained in molecular methods and in conventional microbiology. Thus, the proposed assays are promising for the diagnosis of Salmonella in poultry carcasses, already proved to be faster and more efficient than conventional diagnostics techniques, being of great interest for poultry production, animal, and public health.

Phytase Supplementation under Commercially Intensive Rearing Conditions: Impacts on Nile Tilapia Growth Performance and Nutrient Digestibility.

This study evaluated the effects of phytase supplementation on growth performance and apparent digestibility of Nile tilapia (Oreochromis niloticus) in a commercial fish farm setting. Nile tilapia (6300 male, 57.48 ± 1.04 g) were randomly stocked into 42 floating cages. The experimental design was completely randomized, comprising six treatments and seven replications. Fish were fed five phosphorus deficient plant-based diets with graded levels of phytase supplementation (0, 500, 1000, 1500, 2000 UF kg-1) and an additional diet containing phosphorus supplementation to meet the requirement of this fish species (positive control). After 97 days of feeding, growth performance data were collected and 900 fish (500 ± 10 g) were relocated to 6 floating cages for the digestibility assessment. Quadratic polynomial regression analysis indicated 1537.5 and 1593.2 UF kg-1 as the optimum dietary levels for daily weight gain and feed conversion rate, respectively. Including 2000 UF kg-1 resulted in the higher dry matter, crude protein, energy, and ash apparent digestibility coefficient values. Therefore, phytase supplementation from 1500 to 2000 UF kg-1 is recommended to enhance growth performance and nutrient bioavailability of Nile tilapia reared according to industry practices.