Bacterial contamination of imported poultry feed in Iraq

Transcription

1 PHARMACEUTICAL AND BIOLOGICAL EVALUATIONS October 2016; vol. 3 (issue 5): ISSN Research Article Bacterial contamination of imported poultry feed in Iraq Muhsin L. Al-Musawi, Siham B. Hussein*, Marwa J. Gatoof, Noor Alhuda S. Hanash, Baraa Z. Abdulkareem Iraqi Agriculture Minisitry - Driectorate of Animal Resources - Department of Quality Control on Feed, Baghdad, Iraq *For correspondence Siham B. Hussein, Iraqi Agriculture Minisitry - Driectorate of Animal Resources - Department of Quality Control on Feed, Baghdad, Iraq. sehama106@gmail. com Received: 25 August 2016 Revised: 05 September 2016 Accepted: 15 September 2016 ABSTRACT Objective: This study aimed to evaluation of feed safety because of bacterial contamination of imported poultry feed in Iraq, which causes poultry diseases that often causing death and great economic losses. Methods: Imported poultry feed Samples were collected, cultured in Selenite Cysteine Broth for enrichment of Salmonella (an emphasized organism), used Eosin Methylene Blue agar (EMB) for isolation of Escherichia coli and Isolates were identified according to morphological, biochemical tests and Agglutination test (O&H antiserum for salmonella). Results: The results indicated bacterial contamination of imported poultry feed, among all the samples, we note Gram-negative bacteria in 48 samples of the total samples at (38.4%) included Salmonella in 17 samples at (13.6%), E. coli in 20 samples at (16%), Klebsiella in 3 samples at (2.4%), Shigella in 4 samples at (3.2%) and Proteus in 4 samples at (3.2%). Conclusion: This study concluded that must be assessed the microbial quality of imported poultry feed by manufacturers and health authorities to ensure feed safety because of poultry feed may harbor potential poultry and human pathogens. Keywords: Bacterial contamination, Poultry Feed, Escherichia coli, Salmonella, Proteus, Shigella, Klebsiella spp Introduction The safety and quality of poultry feeds are great subject in developed countries, that feed safety is an essential requirement for all animals. Unsafe feed may also causes great economic losses because of destroying an infected flock of birds. 1 There is a proof that poultry feed is often infected with food-borne pathogenic bacteria. 2 Type of feed processing and storage conditions are the factors that have an effect on the population levels and types of micro-organisms in the feed. Since long time has been known that infections agents can be transmitted to animals through feed contamination, as example, that non-typhi serotype of S. enterica could be 495

2 transmitted to chicken through feed contamination by feces of infected rodents. 3 Poultry feed which is exposed to feces of animals became a reason of human infection through chicken consumption. 4 Poultry feed may contain numerous micro-flora that are obtained from more than one environmental sources such as dust, soil and insects. Material of poultry feed may be inoculated with pathogens during growing, harvesting, processing and storage of feed. 5 Poultry feeds infection with a pathogenic microorganism occur during processing, by handling, mixing of ingredients and exposing the raw materials and finished products. Therefore, poultry diseases and death occur as a result of consumption of contaminated feed. 6 Materials and Methods Preparation of media used The media were prepared according to the manufacturer s instructions on the media labels and autoclaved at 121 C for 15 min. Isolation A total of 125 samples were collected. For Salmonella isolation, 25 g from feed sample was pre-enriched in 225 ml peptone water, mix and incubated, 1ml was transferred to 9ml of Selenite Cysteine Broth and incubated at 37 C for 24 hours. After the incubation period, three differential media, Xylose lysine Deoxycholate (XLD) agar, Hektoen enteric agar and salmonella- shigella agar, were streaked and incubated at 37 C for 24 hours. Salmonella colonies were subcultured on Nutrient Agar and incubated at 37 C for 24 hours. Purification was achieved by sub-culturing and Salmonella suspect colonies were picked up for biochemical tests and agglutination test. For isolation of Escherichia coli used Eosin Methylene Blue (EMB) agar. Identification of the isolates Isolates were recognized, according to morphological and biochemical tests as compared with an identification scheme defined by. 7 Microscopic characteristics Microscopic test appeared as gram negative bacteria when it stained with gram stain. 7 Biochemical characteristics The following tests were used: urease and triple sugar iron (TSI) agar were done to characterize bacteria. 7 Agglutination test: (O and H antiserum for Salmonella): 8 To auto-agglutination, follow the manufacturer s instructions, which is: Add one drop of saline on a glass slide then transfer a small amount of bacterial culture (1 μl loop) onto the glass slide, mix with the drop of saline after it add one drop of antiserum and mixed with a bacterial suspension from a plate or broth on a slide, gently tilt the slide back and forth, this should take 5 to 60 seconds. The presence of granules in the suspension indicates auto agglutination (positive result). Results and Discussion The feed safety is a requirement of Animal Health Organization (OIE) to ensure animal health and welfare as well safety of poultry products for human consumption is a requirement of the World Health Organization (WHO) because of bacterial contamination of poultry feed is an entry pathway of pathogens into the human food supply. 9 A total of 125 feed samples was collected, the isolates were identified according to their microscopic, cultural and biochemical properties to the following: Gram-negative bacteria (48) samples of the total samples (38.4%) included Salmonella in 17 samples at (13.6%), Escherichia coli in 20 samples at (16%), Klebsiella in 3 samples at (2.4%), Shigella in 4 samples at (3.2%) and Proteus in 4 samples at (3.2%), as it is shown in Table

3 Table 1: The number and percentage of each bacterial isolates. Bacteria l isolates No. of positive sample Percentag e of Total Isolation (1) Percentage of Specific Isolation (2) Salmone % 35.4% lla Escheric 20 16% 41.6% hia coli Klebsiell 3 2.4% 6.25% a Shigella 4 3.2% 8.3% Proteus 4 3.2% 8.3% Total % (1) Total Isolation (125) samples. (2) Total specific isolation (48) samples (Gram-negative bacteria) Morphological and microscopical characteristics of bacterial isolates shown as following in Table 2. Table 2: Morphological characteristics and gram stain of bacterial isolates. Bacterial isolates Salmonella Escherichia coli Klebsiella Shigella Proteus Morphological Characteristics Blue- green with black center on XLD agar Deep yellow, round coloured Mucoid, yellow coloured Green coloured Small blue coloured Gram stain single pair or single rod shape rod shape pair Biochemical tests such as (urease and TSI agar) were done to characterize bacteria as shown in Table 3. Table 3: Biochemical Tests for Bacterial isolates. Bacterial TSI agar Urease isolates Salmonella A/K with H2S - Escherichia A/A with gas - coli production Klebsiella A/A + Shigella K/A - Proteus A/A with H2S + Key: =Negative; + = Positive; A = Acid and K = Alkeline. Agglutination test: (O&H antiserum for salmonella): All Salmonella isolates were positive for this test. From the results in a Table 1, we note that the percentage of Bacterial isolates were (48) samples (38.4%) among the total samples, and most isolated Gram-negative bacteria were Escherichia coli (16%), followed by Salmonella (13.6%), Proteus (3.2%), Shigella (3.2%) and Klebsiella (2.4%) as it is shown in Figure 1. Figure 1: The percentage of bacterial isolates. As shown in Figure 1, the percentage of Salmonella isolates was (13.6%) as it is known this bacteria causes salmonellosis, therefore feed has been to be the source of infection to chicken 497

4 and human because of eating chickens fed Salmonella-contaminated feeds. 10,11 It is commonly accepted that Salmonella present in animal feed is a source of animal infection. 12 Among Gram-negative bacteria, the percentage of E. coli isolates was (16%) which is a higher percentage compared with the other bacteria and the percentage of Klebsiella isolates were (2.4%) which is a lower percentage compared with the other bacteria. Both bacteria (E. coli & Klebsiella ) are known pathogens in poultry and humans. E. coli can be contaminants of poultry feed by feces of infected rodents. 13 Poultry feed which is exposed to feces of animals (suspected to harbor E. coli) was determined a reason of human infection via consumption of chickens. 2 The percentage of Shigella isolates were (3.2%), in 2004, first mentioned shigellosis in chicken, characterized by bloody and purulent dysentery in chickens. 14 Shigella pathogenicity is attributed to the organism ' s ability of invading, replicating and spreading in the colonic epithelium to cause the dysentery and other intestinal clinical signs as a result of invading of Virulent Shigella to intestinal epithelial cells. 13 Shigella was isolated from chicken or human is highly similar without unsure evolutionary proof. 15 Shigella isolated from chicken or human have highly similar pathogenicity as well as the possibility of human poultry cross-infection, which is public health significance. 16 The percentage of Proteus isolates were (3.2%), Proteus species cause embryonic death, yolk sac infections, and mortality in young chickens, turkeys and ducks. 17 chickens, which infected with bumble foot, Proteus mirabilis were isolated from the infection position and it has the ability to re-induce the disease when inoculated into the food pad of chickens. 18 In this study, we found it is necessary to evaluate a feed safety by bacteriological examination of imported poultry feed to ensure safety and quality of feed to prevent poultry diseases and great economic losses. Conclusions In this study, we found it is necessary to evaluate a feed safety by bacteriological examination of imported poultry feed to ensure safety and quality of feed to prevent poultry diseases and great economic losses. Funding: No funding sources Conflict of interest: None declared References 1. Ahmed NTH. Bacterial Contamination in Poultry Feed in Khartoum State, master theses, Omdurman Islamic University, Bryan FL, Doyl MP. Health risk and consequences of Salmonella and Campylobacter jejuni in raw poultry. J Food Prot. 1995;58: Wilson JE. Avian Salmonellosis. Vet. Rec. 1948;60: Gangarosa EJ, Barker WH, Baine WB, Morris GK, Rice PA. Aniaml feed as the source of human Salmonellosis. Lancet. 1973;1: Watkins JR, Flower AI, Grumbles LC. Salmonella organism in animal products used in poultry feed. Avian Dis J. 2003;3: Onyeze R, Onah G, Eluke O. Bacterial contaminants associated with commercial poultry feeds in Enugu Nigeria. IJLBPR. 2013;2(3): Holt JG, Krieg NR, Sneath PHA, Staley JT, Williams ST. Bergy's manual of determinative bacteriology. 9th ed. Williams and Wilkins, Baltimore, USA, ISO/TR , Microbiology of food and animal feed Horizontal method for the detection, enumeration and serotyping of Salmonella Part 3 Guidelines for serotyping of Salmonella v , Page 1 of Maciorowski KG, Herrera P, Jones FT, Pillai SD, Ricke SC. Effects on poultry and livestock of feed contamination with 498

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