The reagents and culture media used in this study consist of ingredients of uniform grade and chemicals of analytical grade or the highest purity available. The various analyzes were carried out at Mohammed TAHRI University of Bechar (Algeria) for two months from April 1
st, 2021.
Sampling
The camel meat samples to be analyzed were purchased from the Bechar El Djadid market. The latter is experiencing strong marketing of this product (Table 1).
After the purchase, the sample was put in a refrigeration system (an isothermal cooler) and sent as quickly as possible to the laboratory for its analysis.
Microbiological analysis
The microbiological analyzes were carried out according to the standards established by the American Public Health Association (APHA) (
Salfinger and Tortorello, 2015).
Stock solution and decimal dilution
The prepared stock solution was in the order of 10
-1 with physiological water (0.9%) as the diluent (JORA n.38, 2014). Homogenization of the sample was held in a stomacher sterile blending bag, using a homogenizer (Seward stomacher 400 circulator, England) for 8-10 min (Fig 1).
National regulations (JORA n.39, 2017) require that the sample must be composed of five units to have a representative one (sampling plan n=5). From the stock solution, a series of decimal dilutions were prepared (up to the 10
-5 dilution) with the same diluent.
Microbial parameters analyzed
According to national regulations (JORA n.39, 2017), the microorganisms to be searched belong to the following bacterial groups:
Escherichia coli,
Pseudomonas sp and
Salmonella sp. In addition to these three bacterial parameters, we also analyzed the following parameters: Total Aerobic Mesophilic Flora (TAMF), Coagulase Positive Staphylococci (CoPS),
Listeria monocytogenes, sulfite reducing Clostridia, yeasts and molds.
Search and enumeration of the microbial parameters mentioned above were carried out as follows:
- Search and enumeration of the total aerobic mesophilic flora were carried out by the pour plate technique on PCA (Plate Count Agar). The Petri dishes were incubated at 22 and 30°C for 72 hours.
- The total coliforms (TC) and thermotolerant or Fecal Coliforms FC (
Escherichia coli) were isolated by the pour plate technique on MacConkey agar medium. The Petri dishes were incubated at 30 and 44oC for 24 to 48 hours.
- Search and enumeration of spores of sulfite reducing Clostridia (SRC) by the pour plate technique on Meat-Liver Sulfite Iron Agar in a tube after having exposed the stock solution to heat-shock at 80°C for 10 minutes. The tubes were incubated under anaerobic conditions at 46°C for 24 hours.
- Search and enumeration of
Pseudomonas spp by spread plate technique on cetrimide agar. The Petri dishes were incubated at 37°C for 24 hours.
- Staphylococci were isolated using the spread plate technique on Baird-Parker agar medium. The petri dishes were incubated at 37°C for 24 hours.
- Search for
Listeria monocytogenes was carried out in three stages:
Primary enrichment on half Frase broth, then secondary enrichment on Fraser broth and thirdly, isolation on PALCAM Listeria agar. The Petri dishes were incubated at 37°C for 24 hours.
- Search for
Salmonella sp was carried out in three stages (NF V08-052, 1993):
A pre-enrichment on Buffered Peptone Water (BPW), then, a selective enrichment on Vassiliadis broth (RV) and Selenite Cystine broth (SC) and finally,isolation on Hektoen agar and Salmonella Shigella agar.
The Petri dishes were incubated at 37°C for 24 hours.
- Search and enumeration of yeasts and molds (Fungal Flora‘FF’) were carried out by spread plate technique on Potato Dextrose Agar supplemented with lactic acid. The Petri dishes were incubated at 25°C for 3 to 5 days.
- Search and enumeration of lactic acid bacteria (LAB) were carried out by the pour plate technique (double layer) on MRS (De Man, Rogosa and Sharpe) agar medium incubated at 30°C for 72 hours.
Identification of isolates
The suspected pathogenic isolates namely
Staphylococcus aureus,
Salmonella spp,
Listeria monocytogenes underwent biochemical tests, mainly: Coagulase test, catalase test, urease test, oxidase test and esculin hydrolysis test (
Tille, 2018,
Benyagoub et al., 2018a).
Physicochemical analysis
The physicochemical analysis was limited by four parameters namely: pH measurement, temperature, determination of total solid content and moisture rate, carried out as follows:
- The pH was measured using a multi-parameter pH meter (Adwa AD 1040, Romania) (JORA n.23, 2006). The device was calibrated first.
- The temperature (T°C) of the samples was measured using an electronic thermometer.
- The total solids content (TSC) is the group of all substances which do not volatilize under specific physical conditions.
Moisture content (M%) was measured using the evaporation method of a mass of meat of 10 g at 105°C for 2 to 3 hours. The process was repeated until the weight of the meat sample stabilized. The moisture content was expressed in (%) and calculated according to the following formula:
Where,
m1 and m2 are the masses in (g) of the sample before and after drying, respectively.
Thus, the total solids content (TSC) was calculated by the following formula (JORA n.01, 2006):
TSC (%) = (100 - M)
Where,
M (%): Moisture content in (%).
Interpretation of microbiological analysis
The hygienic quality of the analyzed samples was judged based on the m and M contamination limits (m and M) given by the official journal (JORA n.39, 2017). Microbial parameter results were expressed as Log
10 CFU/g and graphical presentations were plotted as a curve using Origin 2018 software.