Global Journal of Medical Research, G: Veterinary Science and Veterinary Medicine, Volume 21 Issue 2

II. M aterials and M ethods a) Study Area and Sample Collection The present research work was conducted at different retail markets in Khulna district from July 2016 to November 2018 in Quality Feed Lab, Khulna. A total no. of 48 samples collected from the retail markets (Nirala market at Khulna city corporation, Dumuria, and Fultola market) in Khulna district. SL. No. Name of the retail markets No. of collected samples Meat (Thigh + Breast) Total No. of collected samples 01. Nirala 16(8+8) 48 02. Dumuria 16(8+8) 03. Fultola 16(8+8) Year 2021 Global Journal of Medical Research Volume XXI Issue II Version I ( D ) G © 2021 Global Journals 8 Microbiological Evaluation of Poultry Meat Obtained from Different Retail Markets in Khulna District b) Preparation of sample for bacteriological studies Each of the raw meat samples was macerated in a mechanical blender using a sterile diluent as per the recommendation of the International Organisation for Standardisation (ISO, 1995). Ten grams of the thigh meat sample was taken aseptically with sterile forceps and transferred into sterile containers containing 90 ml of 0.1% peptone water. A homogenized suspension made in a sterile blender. Thus 1:10 dilution of the samples were obtained. Later on, using a whirly mixture machine, different serial dilutions ranging from 10-2 to 10-6 were prepared according to the standard method (ISO, 1995). c) Enumeration of TVC For total bacterial count, 0.1 ml of each ten-fold dilution transferred and spread on duplicate PCA using a fresh pipette for each dilution. Then the diluted samples spread as quickly as possible on the surface of the plate with a sterile glass spreader. One sterile spreader used for each plate. The plates then kept in an incubator at 37 0 C for 24-48 hours. Plates exhibiting 30- 300 colonies following incubation. The average number of colonies in a particular dilution multiplied by the dilution factor to obtain the total viable count. The TVC calculated according to ISO (1995). The results of the total bacterial count expressed as the number of organism or colony-forming units per gram (CFU/g) of meat sample. d) Enumeration of TCC For TCC, 0.1 ml of each ten-fold dilution transferred and spread on Mac Conkey agar using a sterile pipette for each dilution. Then the diluted samples spread as quickly as possible on the surface of the plate with a sterile glass spreader. One sterile spreader used for each plate. The plates then kept in an incubator at 37 0 C for 24-48 hours. The growth of the organism confirmed by the appearance of turbidity. Results calculated from MPN tables. e) Enumeration of TSC For total salmonella count, the procedures of sampling, dilution and streaking were similar to those followed in total viable bacterial count. Xylose lysine deoxycholate agar (XLDA) used only in the case of salmonella count. The calculation for TSC was similar to that of the total viable count. f) Enumeration of TCpC For TCpC, 0.1 ml of each ten-fold dilution transferred and spread on the selective blood base agar with 5% sheep or cattle blood. The diluted samples spread as quickly as possible on a 0.45 mm filter placed on blood agar base agar no 2 with a sterile glass spreader. The plates then kept in an incubator at 42 0 C for 24-48 hours. Plates exhibiting 30-300 colonies following incubation. The average number of colonies in a particular dilution multiplied by the dilution factor to obtain the total viable count. The total viable count calculated according to ISO (1995). The results of the total bacterial count expressed as the number of organism or colony-forming units per gram (CFU/g) of meat sample. In young culture, the organism is comma- shaped and S-shaped. In old culture, organisms cling together. Gram (–ve) colonies were round, smooth, and translucent with a dewdrop appearance. g) Cultural and biochemical examination of samples The cultural examination of chicken thigh meat samples for bacteriological analysis done according to the standard method (ICMSF, 1985). The examination followed a detail study of colony characteristics, including the morphological and biochemical properties. To find out different types of microorganisms in chicken thigh meat samples, different kinds of bacterial colonies isolated in pure culture from the plate count agar (PCA), Mac Conkey agar (MCA), blood agar (BA), and xylose lysine deoxycholate agar (XLDA) and subsequently identified according to the methods described by Krieg et al., 1994. The isolated organisms supporting growth characteristics on various media subjected to different biochemical tests such as sugar fermentation test, indole production test, catalase test, coagulase test, methyl-red, and Voges-Proskauer (VP) test. In all cases, standard methods as described by Cowan (1985) followed for conducting these tests. h) Statistical analysis The data on TVC TCC, TSC, and TCpC obtained from the bacteriological examination of meat samples of the poultry carcass collected from Nirala, Table 1: List of the retail market for sample collection