TITLE: Salmonella Typhimurium isolation from specimens
AUTHORS: Angel  Justiz-Vaillant, Suzette E. Curtello

[DESCRIPTION]
<div class = "text-blocks"><div class = "text-block"><span style = "font-style:italic;">Salmonella </span><span>are motile, flagellated rod-shaped Zoonotic pathogens which may survive with or without oxygen and do not absorb crystal violet stain.They are decolourised by alcohol due to their outer lipopolysaccharide membrane and thin peptidoglycan layer.They belong to the family Enterobacteriaceae and is implicated with typhoid fever and food-borne illnesses.This pathogen is associated with intestinal disease which may become fatal and has negatively impact the health of individuals and various economies globally.The poultry industry is most impacted and vulnerable to the onslaught of this pernicious microbe.The Lipopolysaccharide Somatic O antigen, flagellar H and virulent V antigenic structure determines the serotype designate of </span><span style = "font-style:italic;">Salmonella</span><span> species [1]. A wide range of antisera is used in the location of the O antigen associated with the carbohydrate constituent of Lipopolysaccharide and H protein antigen linked with the flagella of the bacteria.</span></div><div class = "text-block"><span>Salmonellosis is an inflammation of the intestinal mucosal lining by </span><span style = "font-style:italic;">Salmonella </span><span>bacteria infiltration resulting in painful symptoms.Both man and animal are vulnerable.Salmonellosis is caused by ingestion of </span><span style = "font-style:italic;">Salmonella </span><span>with food (Wall et al., 1994).The main route of transmission of </span><span style = "font-style:italic;">Salmonella </span><span>microbe is fecal-oral. Ingestion of small concentration of </span><span style = "font-style:italic;">Salmonella </span><span>such as 6 cells with contaminated food or water will lead to salmonellosis infection [2].</span></div><div class = "text-block">References</div><div class = "text-block"><span>1. Varma JK, Marcus R, Stenzel SA, et al. Highly resistant </span><span style = "font-style:italic;">Salmonella</span><span> Newport-MDRAmpC transmitted through the domestic US food supply: a FoodNet case-control study of sporadic </span><span style = "font-style:italic;">Salmonella </span><span>Newport infections,</span><span style = "font-style:italic;"> </span><span>2002-2003.</span><span style = "font-style:italic;">J Infect Dis</span><span>. 2006;194(2):222-230. doi:10.1086/505084.</span></div><div class = "text-block"><span>2. Chen W, Martinez G, Mulchandani A. Molecular beacons: a real-time polymerase chain reaction assay for detecting Salmonella.</span><span style = "font-style:italic;">Anal Biochem</span><span>. 2000;280(1):166-172. doi:10.1006/abio.2000.4518</span></div></div>

[STEPS]
?. The Salmonella Typhimurium isolation was carried out as followed: the exterior of the hen cloaca was first cleaned with a sterilized and moistened cotton balls before application of the cotton tips of each swab applicator.
?. The swabs, caeca, and stomach tissues were immediately placed in a sterile screw-cap test tube containing 9 ml of pre-enrichment broth (buffered peptone water 1%).
?. At least 2.5 g of each type of specimen was dissolved in 250 ml pre-enrichment broth.
?. The inoculated pre-enrichment broth was incubated at 37 ºC for 24 hrs following this incubation it was thoroughly mixed using a vortex mixer.
?. A 1.1 ml aliquot of buffered peptone water 1% was added to 9 ml of enrichment broth (selenite broth, selenite cystine broth, and tetrathionate broth) and further incubated at 37 ºC for 24 hrs.
?. After vortexing 0.16 ml and a 3 mm loopful of inoculum was used to inoculate the differential plating media such as Salmonella Shigella agar selective media, MacConkey agar, bismuth sulphite agar and brilliant green agar that were incubated at 37 ºC for 24-48 hrs. .
?. Following the incubation, as typically, the cultures were examined, and non-lactose fermenting colonies were selected and used to inoculate Kleiger iron agar and urea agar slants.
?. After a further 24 hours incubation period at 37ºC colonies that gave the typical Salmonella Shigella reaction, were inoculated to the routine line of sugars and again incubated.
?. Confirmation was followed by slide agglutination with somatic “O” and flagella “H” antigens of Salmonella.
?. Serological typing of Salmonella Typhimurium was performed.