In the present study, 248 cloacal swabs samples were collected from poultry farms located in different districts across 2 zones of West Bengal, India viz. New alluvial and old alluvial zones. The birds were reared in intensive deep litter system of management. Samples were collected from the age group of 1 week to 5 weeks of broilers. All the collected samples were enriched with peptone water (HiMedia, India) and incubated at 37°C for overnight. The peptone water enriched samples were taken and a loop full of culture was streaked on MacConkey's agar (HiMedia, India) and incubated at 37°C for overnight. Lactose-fermenting, pink single colonies were selected and subcultured on Eosin Methylene Blue agar (HiMedia, India) plates for selective isolation. The characteristic greenish metallic sheen showing colonies were streaked on nutrient agar (HiMedia, India) slants for further morphological and biochemical characterization as per method described by Quinn et al., (1999) . E.coli isolates were serogrouped at National Salmonella and Escherichia Centre, Central Research Institute, Kasauli, Himachal Pradesh, India.
Peptone water
Manufactured by HiMedia
Sourced in India
Peptone water is a microbiological culture medium used for the cultivation and identification of various bacteria. It provides a source of nitrogen and other nutrients essential for bacterial growth. Peptone water is commonly used in diagnostic and research laboratories for activities such as bacterial enumeration, pathogen detection, and biochemical tests.
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Lab products found in correlation
Nessler s reagent, by Thermo Fisher Scientific (2 mentions)
Nutrient agar, by HiMedia (2 mentions)
Macconkey s agar, by HiMedia (2 mentions)
Emb agar, by HiMedia (1 mentions)
Plate count agar, by HiMedia (1 mentions)
Methanol, by Merck Group (1 mentions)
1 1 diphenyl 2 picrylhydrazyl dpph, by Tokyo Chemical Industry (1 mentions)
2 2 azino bis 3 ethylbenzothiazoline 6 sulfonic acid, by Tokyo Chemical Industry (1 mentions)
Potato dextrose agar (pda), by HiMedia (1 mentions)
Nystatin, by Merck Group (1 mentions)
Peptone water, by Merck Group (1 mentions)
Chloramphenicol, by Merck Group (1 mentions)
Eosin methylene blue agar, by HiMedia (1 mentions)
Stuart colony counter, by Cole-Parmer (1 mentions)
Tween 80, by HiMedia (1 mentions)
Ammonium sulfate, by HiMedia (1 mentions)
14 protocols using peptone water
1
Isolation and Serotyping of E. coli from Poultry Farms
2
Microbial Ammonia Production Protocol
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Ammonia production was studied using a standard protocol90 . Bacterial isolate was inoculated in peptone water (Himedia, Mumbai, India) and incubated for 10 days (28 °C, 120 rpm). Test aliquots were withdrawn at different time intervals (48, 120, 160, and 216th h) and centrifuged to remove the pellet (10,000 rpm, 15 min), while 1 ml of the supernatant was mixed with 50 µL Nessler’s reagent (Thermofisher Scientific, New Delhi, India). Yellow to brown coloration in test aliquots depicts positive ammonium production which was quantified spectrophotometrically (OD450) using a standard curve for commercially purchased ammonium sulfate (Himedia, Mumbai, India). All tests were repeated thrice with four replicates per test.
3
Enumeration and Confirmation of E. coli
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Peptone water (Himedia, M028) and Brilliant green lactose bile (BGB) broth (Merck, 736) were used for the enumeration of E. coli using the most probable number technique. Eosin methylene blue (EMB) agar (Millipore, 70186) was used for the confirmation of the presence of E. coli. Then, the conventional biochemical tests were used for further characterization of the isolates [13] .
4
Isolation and Identification of Enterobacteriaceae
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The peptone water (HiMedia, India) containing faecal samples were incubated at 37 0 C for overnight. After that, it was streaked onto MacConkey's agar (HiMedia, India) and again incubated at 37 0 C for overnight. Next day 2-3 rose pink colonies were randomly picked and transferred to EMB agar (HiMedia, India). The plates were incubated overnight at 37 0 C. Next day colonies were observed and a single colony was streaked to nutrient agar (HiMedia, India) slant for further biochemical confirmation.
5
Microbial Profiling of Turmeric and Moin-moin
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The microbiological analyses of the dried turmeric spice and Moin-moin samples were performed on production day and 48 h after production and storage. The dried turmeric powder was assessed for total viable, Enterobacteriaceae and Staphylococci counts. While the moinmoin samples were assessed for total viable, Enterobacteriaceae, coliform and Staphylococci counts on production day and 48 h after production. Briefly, for the microbial load of the spice, 10g of spice was mixed with 90 mL of peptone water (Himedia Lab., India) and further serially diluted up to 10 -4 . Using the pour plate method, 1 mL of last dilution was plated in plate count agar, MacConkey and Baird Parker agar (Lab. M, United Kingdom) for total viable, Enterobacteriaceae and Staphylococci counts respectively (AMPH, 1992). Similar process was followed for the moin-moin samples except Eosin methylene blue agar for Coliform count was also inoculated. Plates were incubated at 37 0 C overnight and enumerated.
6
Antioxidant Potential of Avocado Leaves
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For this experiment, low molecular weight CH (chitin deacetylated ≥ 75%) and Methanol from Sigma–Aldrich Co (St. Louis, MO, USA), Peptone water, Plate count agar (PCA), and Potato dextrose agar (PDA) were bought from HiMedia Laboratories, 507 School House Rd., Kennett Square, PA, USA, 1,1-Diphenyl-2-picrylhydrazyl (DPPH), and 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) were bought from TCI chemicals, Toshima, Tokyo, Japan and fresh AV leaves were harvested from the farm in the College of Agriculture and Veterinary Medicine, at United Arab Emirates University, Al Ain, UAE.
7
Ammonia Production in Fungal Cultures
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Ammonia production was detected in peptone water (Hi-Media). Fungal cultures were inoculated and incubated at 28 °C for 7 days. After incubation, 1.0 ml of Nessler's reagent was added to each tube, and the development of brown to the yellow color indicated the positive test (Cappuccino and Welsh 2018) .
8
Microbial Enumeration and Isolation from Cocoa Beans
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Twenty-five grams of cocoa beans and adhering pulp were added to 100 mL of peptone water (Himedia) and homogenized manually. Serial dilutions in peptone water were prepared and yeasts were enumerated by spreading on YPD agar containing 100 µg/mL chloramphenicol (Merck) in triplicate and AAB were determined by spreading on GYC agar (50 g/L glucose, 10 g/L yeast extract, 30 g/L calcium carbonate, 20 g/L agar, Merck) containing nystatin (0.4%, Merck). Plates were incubated at 30°C for 3 (yeasts) to 5 days (AAB).
After the incubation time, the colonies grown in each medium were counted (mean and standard deviation were calculated). The square root of the number of colonies of each type was re-streaked and purified. The purified isolates were stored at -20°C in YPD broth containing 25% of glycerol.
After the incubation time, the colonies grown in each medium were counted (mean and standard deviation were calculated). The square root of the number of colonies of each type was re-streaked and purified. The purified isolates were stored at -20°C in YPD broth containing 25% of glycerol.
9
Microbiological Analysis of Food Samples
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Upon the completion of storage, samples were aseptically transferred to sterile stomacher bags containing 90 mL of peptone water (Himedia, Mumbai, India). The samples were homogenized for 1 min in a stomacher (Interscience, Saint Nom la Brétèche, France). The mesophilic total plate count (TPC ), yeast and mould (YM ), mesophilic lactic acid bacteria (LAB ), total coliforms (TC ) and Enterobacteriaceae (EN ) were enumerated using Plate Count Agar (30°C for 3 d), Sabaroud Dextrose Agar (SDA , 25°C for 5 d), De Man Rogosa Sharpe Agar (MRS , with an overlay, anaerobically, 25°C for 5 d), Violet Red Bile Agar (VRBA , 32°C for 1–2 d) and Violet Red Bile Glucose Agar (VRBGA , with an overlay at 32°C for 1–2 d), respectively. Meanwhile, PS was enumerated using Pseudomonas Agar supplemented with Pseudomonas CFC (25°C for 48 h) using a spread plate method.
10
Microbial Load Quantification in Food Samples
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After storage, individual samples were aseptically transferred into sterile stomacher bags containing 90 mL peptone water (Himedia, Mumbai, India). Homogenization of the samples was conducted using a stomacher (Interscience, Saint Nom la Brétèche, France) for 1 min. After that, 0.1 or 1 mL of appropriate decimal dilutions was plated in duplicate for Mesophilic total plate count (TPC), yeast and molds (Y&M), mesophilic LAB, and Enterobacteriaceae (EN) via the pour plate method using Plate Count Agar (30 °C for 3 day), Sabaroud Dextrose Agar (SDA, 25 °C for 5 day), De Man Rogosa Sharpe Agar (MRS, with an overlay, anaerobically, 25 °C for 5 day), and Violet Red Bile Glucose Agar (VRBGA, with an overlay at 32 °C for 24–48 h), respectively. A spread plate method was used to enumerate pseudomonads using Pseudomonas Agar supplemented with Pseudomonas CFC, and incubated at 25 °C for 48 h. Subsequently, plates containing 25–250 colony-forming units (CFU) were counted manually using a Stuart® Colony Counter (Cole-Parmer, Eaton Socon, UK) and the resulting data were transformed into log CFU/g [17 (link)].
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