El –kewaiey and mima by ابراهيم القويعى

Alexandria Journal of Veterinary Sciences 2013, 38 (B): 1-7 ISSN 1110-2047, www.alexjvs.com

Incidence of Enterococci in some Heat Treated Poultry Meat Products El –Kewaiey I.A, Omima A.A.S Animal health research instituted Damanhour branch

Key words ProtexinEnterococcus faecium - sea bass fingerlingsgrowth immune status

ABSTRACT: A total of 80 random samples of poultry meat products 20 each of shieshtawook, chicken paneh, nuggets and chicken burger were collected from supermarkets of Al- Bohiera governorate. The samples were examined bacteriologically to estimate their sanitary condition. The mean count of Enterococci were 2.8×10 2 ±1.33102,0.76×10 ±0.28×10, 6.9×102 ±2.09×102_and 4.33 ×102 ±_1.07×102 in shiesh tawook, chicken paneh,nuggets and chicken burger,respectively. The public health hazard of the isolated and identified strains such as Enterococcus fecalis, Enterococcus facium Enterococcus durans, and Enterococcus avian, as well as suggestions to increase the shelf life of the products and protect consumers were discussed.

1-INTRODUCTION the presence of 40% bile salts. E .fecium strains have been able to survive heating to 65ºC for 20 min,71ºC for 10 minutes and 80ºC for 30 min (kearns,et al .1995).Thus,Enterococci may survive some type of food processing, and have been implicated in food spoilage of processed meat (Franz, et al.1999). Enterococci are a part of the natural flora in the intestinal tract of mammals and birds. E.fecalis is found more commonly than E.faecium in human feces (Nobel ,1978).The highest numbers of both species are found in the colon . In addition Enterococcal species can be found in soil, on plants ,and in water. In water they are generally considered as fecal contaminants and belong mainly to the species E.faecium and E.faecalis ,but other species also can be recovered(Niemi,et al.1993). In addition, some strain as well documented as opportunistic pathogens and have been implicated in endocarditis,infant diarrhea and other conditions. However strains are wide spreed in foods.(Eva, 2000). Various studies were carried out on sources of group D Streptococci in food by El-Kateib, (1982), and Kalafalla (1988), who indicated that Streptococcus fecials was predominantly associated human feces . The Enterococci have traditionally been used as indicators of fecal contamination because they are common inhabitants of the human and animal intestinal tract where Streptococcus

In the last decade, there has been a dramatic increase in the consumption of fresh /frozen, precooked/fully cooked chicken-based meat products due to their high nutritional value, lower cost, and convenience and variety aspects for the consumers. Processing of these values added products such as nuggets, chicken burger , chicken paneeh, and shieshta wook is also of benefit to the processor . These types of products require strict hygiene applications throughout The processing chain, since chicken meat/chickenbased products are very susceptible to microbial spoilage and could harbor pathogens (Seran,et al. 2011) Chicken meat and chicken meat products are considered as good sources of animal protein of high biological value which contain all essential amino acid besides they contain many vitamins and minerals which essential for nutrition (Khalifa,et al.2005). Enterococci are Gram-positive cocci that occur singly, in pairs,or as short chains.(Yifan Zhang. 2005). More than 20 Enterococcal species are now recognized. They are facultative anaerobes, tolerant to extremes in temperature, salinity, and pH and are among the most thermotolerant of the non sporulating bacteria (Franz,et al.1999). The optimum growth temperature is 35ºC with a growth range from 10 to 45ºC.They grow in broth containing 6.5% NaCl , and hydrolyze esculin in

Kewaiey and Omima /Alexandria Journal of Veterinary Sciences 2013, 38 (B): 1-7

fecium and durans were widely distributed in nature. With regard to public health significance, Enterococci have been reported to be associated with food poisoning due to consumption of contaminated food products (Batish et al 1980) The presence of pathogenic and spoilage microorganisms in poultry meat and its products remains a significant concern for suppliers, and consumers and public health officials worldwide. Bacterial level of the poultry carcasses used as the raw products, the hygienic practices during manipulation and on time and temperature storage EL-Bassuonys, (2008) Therefore, this work was planned to estimate the Enterococci as indicates of sanitary quality of products and of hazard to consumers and to evaluate the control measures adopted during production and handling of it , where the aim of food hygiene is the production of food which is both safe and clean.

Test tube containing 9ml of sterile from which ten fold serial dilution up to 108 were prepared . Enterococci count (MPN) A series of three tubes each containing 10 ml bromocresol purple azid broth (Hajna, 1951) were inoculated with one ml from the previously prepared serial dilution, the inoculated and control tubes were incubated at 37C for 48 hours. Tubes showing yellow colour were considered as positive and Enterococcal count (MPN/g) was calculated according to Thacher and Clark(1978) Detection of Enterococci A loopful from a positive bromocresol purple azid broth tubes was streaked on Kanamycin asculin azid agar (Mossel et al 1978) and incubated at 37ºC for 24-48 h. Black Colonies surrounded by black holes represented as group D Enterococci Identification of isolated Enterococci Suspected colonies were isolated, purified and identified according to Krieg and holt, (1984)

2. MATERIAL AND METHOD Collection of samples A total of 80 samples of processed poultry meat (20 of each, nugetts,chicken burger, shiesh tawook , chicken panneh )were collected from Damanhour city. These were directly transported to the lab under complete aseptic conditions and were bacteriologically examined

3. RESULT AND DISCUSSION The bacteria of genus Enterococcus spp, also known as Enterococcal form a part of environmental, food and clinical microbiology. They are considered as a indicator of spoilage, or potential pathogenic organisms (Yifan, 2005). Microbiological criteria establish the acceptability of a product based on presence or absence of Microorganisms. Quantity of toxins per mass unit. Volume, area or batch .The microorganisms included in a criterion must be relevant as pathogens indicators or spoilers (Food and agriculture organization (FAO)1997, Commission Regulation (CE)n02073/2005).

Preparation of samples Eleven grams of each samples were taken aseptically and mixed with 99ml of sterile peptone water 0.1% or saline in a sterile homogenizer flask to get a dilution 1:10 . One ml was transferred to a separate sterile

Table 1. Statistical analytical results of Enterococci M.P.N/gm) In examined poultry meat products (n=20) Type of Positive samples products no % Sheish 16 80 tawook Chicken 12 60 paneeh Nugetts 20 100 Chicken burger

Min

Max

Mean x

S.E.M±

2.4×103

2.8×102

1.33×102

4×10

0.76×10

0.28×10

2.4×103

6.9×102

2.09×102

1.5×102

1.1×103

4×102

1.07×102

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From table (1) the nugetts and sheih tawook samples, revealed high percentage of enterococcal count by (M.P.N/gm), which were 100% and 80% , respectively. While, chicken burger and chicken paneeh show lower percentage to some extend as 60% for both respectively. Lilia et al (2007) found the prevalence of Enterococci was significantly higher in summer than in winter and isolated in percent of 64% and 24% of examined burger samples The minimum count of Enterococci in nugetts and shiesh tawook were 7 and 4 (MPN/gm) while the maximum count were 2.4×103 and For each with a mean value 6.92×102 ± 2.09×102 and 2.81×102±1.33×1o2, respectively. Bkheet et al (2006) detect Enterococci from nugetts as 36%, with a mean value of 33.3± 5.3 c.f .u /gm. Nugetts is manufacture from equal portions of breast thigh meat from chicken Table (1) shows also the Enterococcal count of burger was ranged from 1.5×102 to 1.1×103 with a mean value of 4.33×102, which may attributed to the ingredient which enter in the manufacture is minced poultry meat with other ingredients such as spices and salts, and may adulterated by adding of giblets Lower results was detected by Bkheet et al (2006) where they isolate Enterococci from Chicken burger as 48%, with a mean 145±45.5 Seran (2011) found that Enterococcal count in ground chicken meat was 2.92 log c.f.u/g with a mean of 2.77± o.54 and was < 200±o.o in package products which is indicating good personal hygiene and cleaning and disinfection application leading to zero post contamination to the products after

cooking stage. The minimum of Enterococci in chicken paneeh is lower recorded counts as 3 to 4×10 as a maximum count with a mean value 0.76×10±0.28×10 Liete et al (2009) stated 37% of the examined samples presented a Most Probable Number (M.P.N) of Enterococcal spp varying from < 3 to 1.1×105/gm with an average 6.4×103/g .Brag (1982) found that 98 out of 2000 examined samples of various foods contained Enterococci more than103/gm , 13 had counts exceeding 105/gm (the limit for acceptability) 7 were therefore considered unfit for human consumption .But A.P.H.A(1992), some investigators reported that no acceptable levels of Enterococci could be stated because Enterococcus count varied with the products, the holding condition ,time of storage and other factors .In general Enterococci served as a good index of sanitation proper holding condition Leite et al (2009) add although the current laws do not have established standard for Enteococcus species in poultry meat . Enterococci can serve as indicators of fecal contamination and in some cases determine diseases when present in the food. Contaminations with Enterococci in a cooked ready–to–eat chicken meat product and on processing environmental surfaces were determined. Contamination with Enterococci was higher than with E.coli on the surface of all zones, E.coli was not detected in any products; where as 11.1% to 16.7% of samples were adulterated by Enterococci. Contamination was mostly found in the niches of surfaces that were difficult to clean and sanitize, which led to contamination the product transfer conveyor (Suwimon et al 2010)

Table 2. Frequency distribution of Enterococci enumerated by M.P.N/g.of examined poultry meat products (n=20) ShieshTawook

Chicken paneeh

Nuggets

Chicken Burger

Intervals

<3 3 10

10 _ <102

102 _ <103 103

35 6

15 6

8 _

40 _

1 9

6 6

30 30

Kewaiey and Omima /Alexandria Journal of Veterinary Sciences 2013, 38 (B): 1-7

Table 3. incidence of isolated Enterococci strains in the examined poultry meat products (n=20) Isolates Shieshtawook Chicken paneeh Nuggets

no Enterococcus Facalis

Enterococcus Facium Enterococcus avian Enterococcus durans

35 20

_ 4

Chicken Burger

The frequency distribution of Enterococci as in table (2) 3_ < 10 MPN/gm as 3 (15%)and 1(15%) among shiesh tawook and chicken nugetts , also the count 10 < 102 were enumerated in 7(35%), 6 (30), and 9 (45%) respectively,in the three examined products except chicken burger , but the level 102 <103 illustrated in the 4 examined products with a number 3(15%),6(3%),4(20%) and 6 (30%) ,respectively and the level 103 was the highest count in the table and among nugetts and chicken burger 6 (30%) of each and lower in shiesh tawook 3(15%). Keeratipibul (2009) stated that the efficiency of the heat treatment process was sufficient to eliminate Enterococci however the prevalence of Enterococci In cooked chicken after chilling was 2.7% and after slicing and dicing it was 1.3% and 9.3% respectively. These results indicate that contamination occurs after cooking but in the finished products Enterococci prevalence reduced to 1.3%. These results are attributed to the poultry meat products with heat treatment.

4 4

20 20

Suely et al,(2007) they isolated Enterococcus facalis (85%) but lower results were Enterococcus durans (3%)Enterococcus facium (5%), from poultry meat. On the other side Enterococcus facium was isolated in higher percent in chicken burger 6(30%) but lower percent in shiesh tawook 5(30%) and 4(20%) from chicken nugetts but was absent in chicken paneeh. Enterococcus facium could be isolated in a percent 36%,40% from from nugetts and chicken burger by (Bekheet, et al 2007). Enterococci in meat products are taken as indicator of neglected personal hygiene mishandling and bad sanitation .Enterococci survived freezing storage neither increase nor decreased in the number during cold storage at -25ยบC and 30ยบC (Gaafar ,2009) Enterococcus facium is inhibited by low temperature (Aymrich et al.,2000a) Enterococcus avian is absent in shiesh tawook but is higher in chicken nugetts 7(35%), lower in chicken burger 3(15%) and moderate in chicken paneeh 5(20%), respectively. The last identified strain was Entercoccus durans which was higher in chicken paneeh 7(35%), lower in shiesh tawook 4(20%), moderate in chicken nugetts 5(20%) and could not be isolated from chicken burger samples, respectively. The microbiological quality of chicken meat can be determined by its sanitary conditions, contamination during processing, storage condition,

Table (3) shows the identified Enterococcal strain as Enterococcus facalis was the highest isolated strain from shiesh tawook 7 (35%) and lowest in chicken burger 3 (15%) but could isolated from nugetts as 4(20%) and absent in chicken paneeh samples respectively . Nearly lower and higher results could be isolated by Bkheet, et al (2007) who isolate Enterococcus facalis from nugetts and chicken burger as 4% and 20%, respectively. Higher results were reported by

Kewaiey and Omima /Alexandria Journal of Veterinary Sciences 2013, 38 (B): 1-7

transportation and commercialization however, contamination occurs easily in the first stage of slaughtering Galhardo, et al 2006) Enterococci are also known to be better indicator of hygiene and fecal contamination than coliform bacteria particularly in heat- treated products (Seran,et al 2011). Factors such insufficient microbial quality of meat and non-meat ingredients improper hygienic applications prior /during and post processing , inefficient heat treatment could result products potentially hazardous to consumer . Also shorter shelf –life and recalls cause economic losses to the producer. The widespread use of antimicrobials in human and veterinary medicine, as well as in animal production has accelerated the development of drug resistance in a variety food safety and public health concern. But their mechanisms of antimicrobial resistance are relatively less clear than those in Gram –negative pathogens Conclusion The presence of Enterococci species (Enterococcus fecalis Enterococcus facium , Enterococcus durans Enterococcus avian) in heat treated poultry meat products could attributed to improper hygienic measure, or using of bad quality poultry meat or additives

the washing , chicken are submitted to evisceration , which can be automated or not. In this step viscera tearing and the direct contact with contaminated surface must be avoided. The automated stage involves cloacae extraction, abdomen cutting, and evisceration, after this stage, sanitary inspection occurs and viscera are manually removed, and the edible giblets(heart, liver and gizzard) are separated to be chilled .Windpipe and crop are removed mechanically, and carcasses are finally wash both external and internally, before passing through the pre-chilling systemAmal Fahmy A.M (1995): Incidence and significance of Enterococci in some meat products thesis for degree of master Meat Hygiene Faculty of vet med university of Alex.

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Recommendation and suggested measure - Implementation of HACCP (Hazard Analysis and critical control points) in poultry industry is extremely important, because it involves the constant monitoring of all steps of the process, aiming the food safety of final products. Indusries must implement this food safety program to serve both internal and external markets . Laboratory control of various step of production - The poultry meat must be free from any dirts . -Mincing machines must thoroughly washed before and after mincing -Maintaining clean water supply -Disinfecting the environment periodically and sanitary disposal of water -Air filtration system should applied in processing plants as air is an important source of contamination. -Educational program should be given for processor and handlers about the quality of the products and ensure maximum of safety to consumers. -Hygienic precautions during transportation and storage should be adopted Before evisceration process, carcasses must be washed in shower with pressured water, since the whole carcass must be washed including feet .After

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