READY-TO-EAT MEAT AND POULTRY PIES AS A SOURCE OF POTENTIAL PATHOGENS IN ASSUIT CITY

M. EBRAHEEM, LUBNA; M. MOHAMED, GHADA

doi:10.21608/avmj.2012.172809

READY-TO-EAT MEAT AND POULTRY PIES AS A SOURCE OF POTENTIAL PATHOGENS IN ASSUIT CITY

Document Type : Research article

Authors

¹ Animal Health Research Institute, Assiut Regional Laboratory.

² Animal Health Research Institute, Assiut Regional Laboratory

10.21608/avmj.2012.172809

Abstract

Atotal of 80 samples of pies, 40 each of meat and poultry pies were collected from different restaurants of Assiut city and analyzed bacteriologically (count of aerobic bacteria, coliform, faecal coliform Staph aureus, E.coli and isolation of Staph aureus, Clostridium perfringens, Escherichia coli and Listeria monocytogenes). The mean values of aerobic plate count and Staph. aureus counts were 8×10⁴ and 5X10cfu/g of examined meat pies while that of poultry pies were 8.3 x 10⁵and 6 x 10² cfu/g respectively. A signifhcant difference in such counts was observed between the two types of pies examined. Regarding the MPN of coliform and faecal coliform, the mean values were 1.1X10 and 0.5X10 for meat pies whereas the corresponding values for poultry pies were 0.7 X10 and 0.5 X10 respectively. However, Staph. aureus, Clostridium perfringens, Escherichia coli and Listeria monocytogenes were isolated from 82.5,17.5,7.5 and 15% of the examined meat pies while poultry pies contained such organisms in 77.5,12.5,12.5 and 22.5% of the examined samples. The public health importance of recovered microorganisms as well as some recommended measures for improving the quality of such products were discussed.

Keywords

Full Text

Ready-To-Eat Meat and Poultry Pies as a source of potential pathogens in Assuit city

Lubna M. Ebraheem and Ghada M. Mohamed

Animal Health Research Institute, Assiut Regional Laboratory.

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Abstract

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Received at: 1/3/2012

Accepted: 9/4/2012

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Key words: Meat pies, poultry pies, potential pathogens.

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فطائر اللحوم والدجاج المعدة للأکل کمصدر محتمل لمسببات الأمراض في مدينة أسيوط

لبنى محمد ابراهيم , غادة محمد محمد

لقد تم جمع 80 عينة من الفطائر بواقع 40عينة من فطائر اللحم و40 اخرى من فطائر الدجاج من مطاعم مختلفة بمحافظة أسيوط واجرى الفحص البکتريولوجى لها حيث تم تحديد العدد الکلى لکل من: الميکروبات الهوائية, ميکروبات القولون, القولون البرازية, ميکروب المکور العنقود الذهبى والايشريشية القولونية. وقد تم عزل کل من الميکروب المکورالعنقودي الذهبي والکلوستريديم بيرفرنجنز والايشريشية القولونية والليستيريا مونوسيتوجين حيث وجدت ان المتوسطات للميکروبات الهوائية والميکروب المکورالعنقود الذهبى8 ×⁴10, 5X10/جرام وذلک فى فطائر اللحوم بينما کانت هذه المتوسطات فى فطائر الدجاج 8.3× ⁵10 و6×²10/جرام بالترتيب. ولقد وجد اختلاف معنوى بين هذه المتوسطات فى نوعى الفطائر التى فحصت. اما بالنسبة للعدد الکلى المحتمل للميکروب القولونى والميکروب القولونى البرازى کانت المتوسطات لها1.1×10و0.5×10 بالنسبة لفطائر اللحوم بينما کانت هذه المتوسطات فى فطائر الدجاج 0.7×10و0.5×10بالترتيب. وتم عزل الميکروب المکورالعنقود الذهبى والکلوستريديم بيرفرنجنز والايشريشية القولونية والليستيريا مونوسيتوجين من فطائر اللحوم بالنسب الاتية: 82.5, 17.5 ، 7.5، 15% بالترتيب بينما عزلت هذه الميکروبات من فطائر الدجاج بالنسب الاتية: 77.5، 12.5 ، 12.5 , 22.5%بالترتيب. وقد تمت مناقشة الأهمية الصحية لهذه الميکروبات ومدى خطورتها على الصحة العامة کذلک الطرق المقترحة للحد من هذه الخطورة.

Introduction

Ready-to-eat foods could be raw or cooked, hot or chilled and can be consumed without further heat treatment (Tsang, 2002).

Examples of such foods included meat pies and poultry pies. This situation however has resulted in more ready to-eat-foods taken outside home, this food vendor services become on the increase and responsibility for good manufacturing practices of food such as good. The sanitary measures and proper food handling must be improved, because venders can transfer diseases between families through ready-to-eat foods (Musa and Akande, 2002).

Mishandling in food service establishment can contribute significant outbreaks of foodborne disease (Frazier and Westhoff, 2001), also these outbreaks are caused by foods that are contaminated intrinsically (Torok et al., 1997). Because of modern processing methods, handling and distribution, it takes longer for food to reach the table, and it is more likely to be contaminated with microorganisms. (Marriot, 1997).

Staph. aureus, Clost. perfingens and Escherichia coli are bacteria that cause food poisoning. Food infection is the second type of foodborne illness. It is caused by eating food that contain certain types of live bacteria which are present in the food. Once the food is consumed, the bacterial cells themselves continue to grow and illness can result, Listeria is a good example of foodborne infection. Each year, thousands of individuals suffer from the discomfort and pain resulting from foodborne illness. True food poisoning or food intoxication caused by eating food that contains a toxin or poison due to bacterial growth in food (Estes et al., 2003).

It is extremely difficult to eliminate contamination of food completely, but it is easier to control further multiplication of pathogens and stop the forms that produce toxins (Bryan et al., 1991). The bacteria which produced and excreted the toxic waste products into the food may be killed, but the toxin they produced causes the illness or digestive upset to occur (Estes et al., 2003).

Prevention of foodborne infections is based on: pathogen-free food production, hazard control in food processing, surveillance for foodborne illness, and safe food handling by consumers and food-service workers (Doyle, 1993). The practices developed by professionals to ensure the safety of food production and processing are based primarily on knowledge of microbiology (Altekruse et al., 1996).

Since the rules of how to avoid food poisoning are known, the problem in mass catering is enforcement which must include not only proper equipment and installations but also training of personnel (Teuber, 1992).

As there is scarce of information about the bacterial status of pies in AssiutCity therefore the aim of this study was focused to assess the bacteriological quality of meat and poultry pies in Assiut governorrate.

Materials and Methods

To ascertain the levels and types of bacteria present in meat and poultry pies, 40 samples of each ready-to-eat products were obtained from randomly selected restaurants of AssiutCity and examined on the day they were collected for:

1- Total colony count (APC): APHA (1992).

2- Coliform count, Faecal coliform count, E.coli count (MPN/g): AOAC (1990).

3- Staph. aureus count: APHA (1992).

4- Isolation of Staph aureus: Feingold and Martin (1982).

5- Isolation of Listeria monocytogen: Oxoid Manual (1990).

6- Isolation of Clostridium perfringens: Beernes et al. (1980).

7-Isolation of E.coli: AOAC (1990).

All isolates were identified morphologically using staining reaction (APHA, 1992) and motility test (Baron et al., 1994), as well as, biochemically using catalase, coagulase and triple sugar iron (TSI) agar test (Baron et al., 1994), citrate utilization, indole production, methyl red, urease and voges-Proskauer tests (Koneman et al., 1992), nitrate reduction test (Cowan and Steel, 1974), sugar fermentation reaction (APHA, 1992), Christine-Alkine- Munch- Peterson (CAMP) test (Herrera, 2001). For further confirmation of L.monocytogenes the isolates were inoculated into 10% aqueous stock solution of Mannitol, L.Rhammose and D.xylose (Collee and Miles, 1989).

RESUTS

Table1: Statistical values of aerobic plate count / gram of the examined meat and poultry pies.

	No of exam samples	No.of + ve samples	Minimum	Maximum	Mean	S.E	P. value
Meat pies	40	40	9 x 10³	2.9 x 10⁵	8 x 104	± 1 x 10⁴	< 0.01*
Poultry pies	40	40	2.6 x 10⁴	6.9 x 10⁶	8.3 x 10⁵	± 4.3 x 10⁵	< 0.01*

Table 2: Statistical values of Sstaph aureus count / gram of the examined meat and poultry pies.

	No of exam samples	No.of+ ve samples	Minimum	Maximum	Mean	S.E	P. value
Meat pies	40	31	1 x 10	2 x 10²	5 x 10	1.4 x 10	< 0.01*
Poultry pies	40	30	4 x 10	3 x 10³	6 x 10²	5 x 10²	< 0.01*

Table 3: Statistical values of coliform count (MPN) / gram of the examined meat and poultry pies.

	No of exam samples	No.of+ ve samples	Minimum	Maximum	Mean	S.E	P. value
Meat pies	40	40	3.6	4.3 x 10	1.1 x 10	± 0.1 x 10	N.S
Poultry pies	40	38	3.6	23	0.7 x 10	± 0.1 x 10	N.S

Table 4: Statistical values of faecal coliform count (MPN)/gram of the examined meat and poultry pies.

	No of exam samples	No.of+ ve samples	Minimum	Maximum	Mean	S.E	P. value
Meat pies	40	36	3.6	1.1 x 10	0.5 x 10	± 0.04 x 10	N.S
Poultry pies	40	35	3.6	1.5 x 10	0.5 x 10	± 0.08 x 10	N.S

N.S: non significant

* : Significant

Table 5: Frequency distribution of meat and poultry pies based on their E.coli count (n=40 of each).

Interval	Meat pies		Poultry pies
Interval	No.	%	No.	%
>3	38	95	37	92.5
3->6	1	2.5	3	7.5
6->9	1	2.5	-	-

Table 6: Incidence of the isolated organisms from the examined meat and poultry pies.

Organisms	Meat pies			Poultry pies
Organisms	No. of exam. samples	No. of +ve samples	%	No. of exam. samples	No. of +ve samples	%
Staph aureus	40	33	82.5	40	31	77.5
C.perfrengenes	40	7	17.5	40	5	12.5
E.coli	40	3	7.5	40	5	12.5
L.monocytogenes	40	6	15	40	9	22.5

Discussion

Regulatory agencies and industrial quality assurance regularly examine foods or ingredients for microorganisms or their metabolic productsthat may indicate: (1) The possible presence of a pathogen or harmful toxins, (2) The possibility that faulty practices occurred during production, processing, storage and distribution, and for (3) The suitability of a food or ingredient for a desired purpose (NAS, 1985).

The standard plate count is one of the most common tests applied to indicate the microbiological quality of food. High count may indicate the product may have been prepared unhygienically or stored inappropriately (ICMSF, 2001).

The results recorded in Table 1 reveal that all the examined samples (100%) contained arobic bacteria in numbers varied from 9 x 10³ to 2.9 x 10⁵ with a mean value of 8 ± 1 x 10⁴ CFU/g, in meat pies while in poultry pies the numbers varied from 2.6 x 10⁴ to 6.9 x 10⁶ with a mean value of 8.3 ± 4.3 x 10⁵ CFU/g. There was a comparable difference (P < 0.01) between the mean total microbial counts between meat pies and poultry pies.

The high microbial counts may be due to meat offers a rich nutrient media for microbial growth (Phillips, 2003). Also this is an indication of recontamination in food handling hygiene techniques starting from the processing of the raw material to the finished product (Ikeme, 1990; Ojeibun, 1994). Higher counts of aerobic bacteria in meat pies were also enumerated by Alexander and Tittiger (1971), and a lower figure 3 x 10³ – 5 x 10³ CFU/g was reported by Yah clarence et al. (2009).

Table 2 showed the statistical values of Staph aureus (CUF/g) of the examined meat and poultry pies, the count ranged between 1 x 10¹and 2 x 10² with a mean value of 5 ± 1.4 x 10¹ CFU/ g meat pies and ranged between 4 x 10¹and 3 x 10³ with a mean value of 6 ± 5 x 10² CFU/g in poultry pie, that a significance difference was apparent between them (P < 0.01). Higher counts of Staph aureus in meat pies were recorded by El-Gohary (1994) and Yah Clarence et al. (2009).

The significance difference (P < 0.01) which showed in both of total microbial counts and Staph aureus counts between meat pies and poultry pies could be attributed to the ingredients used and the processing difference.

Results of coliform count (MPN/g) indicated in Table 3 verify that the minimum value of coliform in meat pies was 3.6, the maximum value was 4.3 x 10¹ and the mean value was 1.1 x 10¹ ± 0.1 x 10¹, while in poultry pies the minimum, maximum and mean values were 3.6,23 and 0.7 x 10¹ ± 0.1 x 10¹, respectively.

According to the data summarized in Table 4, the faecal coliform count (MPN/g) varied from 3.6 to 1.1 x 10¹with a mean value of 0.5 x 10¹ ± 0.04 x 10¹ in meat pies and this count varied from 3.6 to 1.5 x 10¹ with a mean value of 0.5 x 10¹ ± 0.08 x 10¹ in poultry pies.

Regarding meat pies, two samples had E.coli where the MPN for each lied between 3->6 and 6->9/g whereas poultry pies revealed the presence of E.coli in 3 samples only where the MPN lied between 3->6/g as recorded in Table 5.

There was no significant difference between meat pies and poultry pies in coliform and faecal coliform counts as recorded in Tables 3 and 4.

Results given in Table 6 revealed that 82.5% and 775% of Staph aureus were isolated from meat and poultry pies respectively, this percent was higher than that obtained by Alexander and Tittiger (1971) (22.7%) in meat pies. Staphylococcus is a true food intoxication organism. It produces a heat stable toxin when allowed to grow for several hours in foods such as chicken pies. This bacterial growth may not cause any off color, odor, or textural or flavor changes, but the toxin will be secreted into the food. This organism grows best at body temperature (36.6 ^oC), but it can grow over the much wider range of 10 ^oC to 46.1^oc. the best prevention of staph food poisoning is to properly store food and reduce the temperature below 40 degree F within four hours after preparation of serving. In order for Staph to grow and produce toxin, it must have sufficient time, approximately two to four hours. Therefore, it is important to cool or heat foods through the danger zone of 4.4^oC to 60^oC as rapidly as possible.

C.perfringens was found in 7 samples of meat pies (17.5%) and in 5 samples of poultry pies (12.5%) (Table 6). Alexander and Tittiger (1971) reported that clostridia were found in low numbers in some samples of meat pies.

C.perfriongens can grow over a wide range of temperature, but grows very slowly at low temperatures, these bacterial spores will germinate and grow best at temperatures between 37.7◦ C and 47.2◦C.

Many foods such as meat and poultry may carry the organism, but the mere presence of C.perfringens in food is not enough to cause illness. Millions of growing cells are needed. The prevention of growth of this organism is best accomplished by following the standard food service practices of rapidly chilling prepared foods in shallow containers and keeping cold food cold and hot food hot, also reduce the level of contamination by keeping all work areas clean and sanitary (Estes et al., 2003).

Results given in Table 6 revealed that 7.5% of E-coli were isolated from 3 out of 40 samples of meat pies and 12.5% were isolated from 5 out of 40 sampels of poultry pies. Alexander and Tittiger (1971) reported that the incidence of samples (Meat pies) containing coliform was high (81.6%), while Yah Clarene et al. (2009) cited the count of E.coli in meat pies as 2 x 10³ CFU/g.

According to Edema et al. (2001), Okonko et al. (2008 a, b) the presence of E.coli in food is an indication of faecal contamination of the water sources that were utilized in the processing of these food products. Also Edema et al. (2005) reported that biological contaminants of bacterial origin present as major cause of foodborne disease given rise to acute to chronic illnesses such as E.coli gastroenteritis, brucellosis and campylobacteriosis.

Listeria monocytogenes was detected in our study in 15% of meat pies and in 22.5% of poultry pies (Table 6).

This organism is a food infection bacteria gaining in public awareness as a safety problem in food products. The general growth conditions required are oxygen, temperatures ranging from 2.7 C° to 40 C° and a pH range of 5.6 to 9.8, since Listeria can grow at refrigerated temperatures. The organism is generally destroyed by heat treatment, 76.6 °C for 15 seconds. Proper personal hygiene, good sanitation, proper cooking and preventing cross contamination of raw and cooked food are the best control measures known to date (Estes et al., 2003).

Thus to safeguard against the risks of detected microorganisms, there is need to educate and advocate for good manufacturing practices among food processors and food vendors.

Betty and Richard 1994, said that food poisoning / illnesses are entirely preventable by practicing good sanitation and food handling techniques.

In conclusion, a mong the requirement of any food to be of good sanitary quality, it must be free from hazardous microorganisms, or these present should be at a safe level. Therefore, standards for composition and bacterial content are now adopted by nearly all countries. so that public may be assured of a safe healthful product.

The information given by the achieved results proved that most of the examined meat and poultry pies contained valiable numbers and types of microorganisms which may be responsible for inferior quality of the product and increase the risk of public health. Therefore, food safety standards should be applied and Hazard Analysis Critical Point (HACCP) should be applied throughout processing and distribution of the product.

References

Alexander, D.C. and Tittiger, F. (1971):Bacteriological studies on meat pies and frozen prepared dinners. Can. J. Comp. Med. Vo. 35: 5-11.

Altekruse, S.F; street, D.A; Fein, S.B and Levy, A.S. (1996): Consumer knowledge of foodborn microbial hazards and handling practices. J. Food Prot. 59, 3: 2287-294.

AOAC "Association of Official Analytical Chemists" (1990):Official Methods of Analysis of the Association of Officical Analytical Chemists. 15^th Ed. Inc. USA. AOAC.

APHA "American Public Health Association": (1992):Compendium of Methods for the Microbiological Examination of Foods 3^rd Ed. Washington, D.C.USA. APHA.

Baron, Ellen, J. O.; Perteson, L. R.; Finegold and Sydney, M. (1994):Bailey and Scott's Diagnostic Microbiology. 9^th Ed Shanahan J.F. (edit). Mosby year Book, Inc.

Beernes, H.; Romond, C.; Lepage, C. and Crquelion, J. (1980): A direct method for the enumeration of Clostridium perfringens in foods and faeces. World Congress foodborne infections. Berlin (West).

Betty, C.H. and Richard, J.B. (1994):Bacterial causes of food poisoning in food poisoning and hygiene, 8^th Ed. Edward Annod. Publishing Limited London PP. 250-257.

Bryan,F.L.; Fukunaga, I.; Tsutsumi, S.; Niyashiro, L.; Kagawa, D.; Sakai, B.; Mastuura, H. and Ormarura, M. (1991): Hazard analysis of Japanese mixed lunches (bento). J. Environ. Health 54: 29-32.

Collee, J.G. and Miles, R.S. (1989):Tests for identification of bacteria Mackie and McCartney Practical Medical Microbiology, J.G. Collee, J.P. Duguid, A.G. Fraser and B.P. Marmion (eds.) Vol. 11, 13 Ed, Churchill living stone Edinburgh, London, PP: 141-159.

Cowan, S.T. and Steel, K.J. (1974):Manual for Identification of Medical Bacteria, 2^nd Ed. Campridge, Campridge Unif. Press.

Doyle, M.P. (1993): Reducing foodborne disease-hat are the priorities? Nutr. Rev., 51: 345-34.

Edema, M.O.; Omemau, A.M. and fapetu (2001):Microbiology and physicochemical analysis of different sources of drinking water in AbeokutaNigeria. Nig. J. Microbiol. 15 (1): 57-61.

Edema, M.O.; Atayese, A.O. and Idowu, A.O. (2005):Microbiological quality of microwave processed foods. In: the Book of Abstract of the 29^th Annual conference and general meeting on microbes as agents of sustainable development. Organized by Nigerian Society for Microbiology (NSM), UNAAB, Abeokuta from 6-10^th Nov. P. 17.

ElGohany, A.H. (1994):Susage and minced meat as a source of food poisoning microorganisms to man. Assiut. Vet. Med. J. 30: 146-215.

Estes, R.; George, S.; William, H. and Typor, P.T. (2003):Preventing food poisoning and food infection the University of Georgia, College of Agricultural and Environmental Sciences cooperative extension service.

Feingold, S.M. and Martin, W.J. (1982): Bailey and Scott Diagnostic. Microbiology. 6^th Ed. C.V. Mosby. Co. St. Louis, Toronto, London.

Frazier, W.C. and Westhoff, D.C. (2001):Food Microbiology, Tata McGrow Hill.

ICMSF (2001): "International Commission on Microbiological Specifications" for Foods. Guidelines for the microbiological examination of ready-to-eat food. December: 18-23.

Herrera, A.G. (2001): Listeria monocytogenes. In food Microbilogy Protocols. Spencer, J.F.T. and Ragout de Spencer (edit). Humana Press Inc. Totowa, New Jersy.

Ikeme, A.L. (1990):Fermental sausage-dry and semidry. Meat Science and Technology 1^st Ed. The African fep Publisher Llimited Nigeria P. 210.

Koneman, E.W.; Schrechenberger, P.C.; Allen, S.D.; Winn, W.C. and Janda, W.M. (1992): Color Altas and Textbook of Diagnostic Microbiology, 4^th Ed. Winters, R. (edit). J. B. Lippincott company, Philadelphia.

Marriott, N.G. (1997): Essentials of food sanitation. Chapman and Hall, 115 Fifth Avenue, New York, NY10003.

Musa, O.L. and Akande, T.M. (2002):Effect of health education intervention or food safety practice among food vendors in llorin. Sahel Med. J. 5: 120-124.

NAS"National Academy of Science"(1985): An Evaluation of the Role of Microbiological Criteria for Foods and Food Ingredients. NationalAcademy Press. Washington, D.C.

Ojeibun, F.I. (1994):Isolation of Staphylococcus aureus and other bacteria from some ready to eat foods sold in Benin City. B. Sc.HonThesisUniversity of Benin, Benin City P. 100.

Okonko, I.O.; Adejoye, O.D.; Ogunnusi, T.A.; Fajobi, E.A. and Shittu, O.B. (2008 a):Microbiological and physicochemical analysis of different water samples used for domestic purposes in Abeokuta and Ojota, Lagos State, Nigeria, Afr. J. Biotechnol. 7 (3): 617-621.

Okonko, J.O.; Ogunjobi, A.A.; Adejoye, O.D.; Ogunnusi, T.A. and Olasogba, M.C. (2008b):Comparative studies and microbial risk assessment of different water samples used for processing frozen sea-foods in Ijoraolopa, LagosState, Migeria. Afr. J. Biotechnol. 7 (16): 2902-2907.

Oxoid Manual (1990):Listeria species and listeriosis 6^th Ed. Unipath Limited, Waderoad, Basingstoke, Hampshire, England.

Phillips, M. (2003):Analysis of Microbial Hazards related to time/ temperature control of foods for safety comprehensive review in Food Science and Food Safety 2: 33-35.

Teuber, M. (1992): Special problems of mass catering 3rd World Congress Foodborne Infection and Intoxication, Vol.II, 16-19 June, Berlin.

Torok, T.J.; Tauze, R.V.; Wise, R.P.; Livengood, J.R.; Sokolow, R. and Manvans, S. (1997):A large community outbreak of salmonellosis caused by international contamination of a restaurant salads/ bars. J. Am. Med. Assoc. 278 (8): 389-395.

Tsang, D. (2002):Microbiological guidelines for ready to eat food. Road and Environmental Hygiene Department Hong. Kong PP. 115-116.

Yah Clarence, S.; Nwinyi Obinna, C. and Chinedu shalom, N. (2009):Assessment of bacteriological quality of ready to eat food (Meat pie) in Benin City metropolis, Nigeria, African Jounral of Microbiology Research. 3 (6): 390-395.

References