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Detection of Salmonella SPP

The document outlines a Standard Operating Procedure (SOP) for the detection of Salmonella spp. in food and water using various enrichment and confirmation methods. It details the objective, scope, materials, equipment, procedure, quality control, reporting, and safety measures involved in the testing process. This SOP is applicable to various food types and emphasizes the importance of proper laboratory practices.

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44 views3 pages

Detection of Salmonella SPP

The document outlines a Standard Operating Procedure (SOP) for the detection of Salmonella spp. in food and water using various enrichment and confirmation methods. It details the objective, scope, materials, equipment, procedure, quality control, reporting, and safety measures involved in the testing process. This SOP is applicable to various food types and emphasizes the importance of proper laboratory practices.

Uploaded by

akolawalafood
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
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ISO9001:2015

AKOLAWALA FOOD PROCESSING ASSOCIATION


Plot No. P-60, Akola Growth Center, Behind Ashish Hotel, MIDC Phase IV, Akola-4441004
MH.

LABORATORY MANAGEMENT SYSTEM


SOP For Determination of Total Ash
Document Number Revision no: Issue No:
Revision Date: Effective Date

Detection of Salmonella spp.

FSSAI Code: 15.016 / 15.017 | Purpose: Detection and confirmation of Salmonella in food
and water

1. Objective
To detect the presence of Salmonella spp. in food and water samples using selective
enrichment, differential plating, and confirmation methods.
2. Scope
Applicable to all food types, particularly meat, poultry, dairy, seafood, and ready-to-eat foods,
and potable water.

3. Materials and Media


• Pre-enrichment: Buffered Peptone Water (BPW)
• Selective enrichment:
o Rappaport–Vassiliadis Soya Peptone Broth (RVS)
o Tetrathionate broth (TT)
• Plating media:
o Xylose Lysine Deoxycholate (XLD) agar
o Hektoen Enteric agar
• Biochemical media: Triple Sugar Iron (TSI), Lysine Iron Agar (LIA)
• Serological reagents: Salmonella O and H antisera

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ISO9001:2015

4. Equipment
• Incubators at 37 °C and 42 °C
• Water bath (optional)
• Inoculation loops, sterile pipettes
• Colony counter
• Slide agglutination kit

5. Procedure
A. Pre-enrichment:
• Add 25 g of sample to 225 mL BPW in sterile bag.
• Homogenize and incubate at 37 °C for 18–24 hours.
B. Selective enrichment:
• Transfer:
o 0.1 mL to 10 mL RVS broth (incubate at 42 °C for 24 hours)
o 1 mL to 10 mL TT broth (incubate at 37 °C for 24 hours)
C. Plating:
• Streak loopful from each enrichment broth onto:
o XLD: Look for red colonies with black centers
o Hektoen: Green/blue colonies with black centers
• Incubate at 37 °C for 24 hours

6. Confirmation
A. Biochemical Tests:
• Pick 2–5 presumptive colonies
• Inoculate into:
o TSI: Alkaline (red) slant / acid (yellow) butt with H₂S
o LIA: Purple butt, H₂S production
B. Serological Test:
• Perform slide agglutination using Salmonella antisera (O and H antigens)

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ISO9001:2015

7. Quality Control
• Positive control: Salmonella Typhimurium
• Negative control: E. coli

8. Reporting
• Report as:
o “Salmonella spp.: Detected / Not Detected in 25 g or 25 mL sample”
• Record complete steps, media used, incubation, and confirmation.

9. Safety and Waste Disposal


• BSL-2 handling precautions
• Autoclave all plates, tubes, and enrichment cultures
• Disinfect surfaces with 1% sodium hypochlorite

Prepared By: Quality Analyst Approved By: Laboratory In charge


Verified By: Techinical Manager Page no: 1 of 2

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