Salmonella Enteritidis in Layer Farms of Different Sizes Located in Northern China: On-Farm Sampling and Detection by the PCR Method

Abstract

ABSTRACT Salmonella enterica subspecies enterica serotype Enteritidis (SE) has caused foodborne infections over decades. It is transmitted mainly from contaminated eggs to humans. SE is commonly present in layer houses, and closely interacts with environmental factors. The objective of the present study was to develop a viable PCR method to identify SE in environmental samples collected in layer farms of different sizes, and to evaluate SE contamination status in four main egg-production provinces of northern China. After specificity retrieval using Primer-BLAST against the NCBI database, three SE specific oligonucleotide primers were selected as candidate primers. The primers targeting Prot6e gene were adopted and primers targeting Sdf I were also selected to validate the results, after testing eight different types of pooled poultry environmental samples (overshoe, air, drinking nipple, feed, egg collection belt, eggshell, air inlet, and air outlet) by PCR. A PCR detection limit of 1 CFU/mL was determined using cell lysates from pure cultures. Testing time was less than 48 h. On-farm samples were collected from two layer farm sizes (one housing more than 50,000 layers, and the other, less than 50,000 layers) in each province. The applied PCR method was shown to be simple, inexpensive and effective for screening SE in a large amount of farm samples. The study identified only one SE-positive farm, which a large farm and where nine samples were found to be contaminated with SE: drinking nipples (3), egg collection belt (1), air inlet (1), air (1), overshoe (1) and eggshell (2).