International Food Safety Conference th 5 Damanhour University October 2018 th , 13 Saturday 337 Effect of Carica Papaya seeds, seeds and leaves extract on Staphylococcus aureus and Escherichia coli in minced meat and on meat quality Ebeed Abd Elatty Saleh 1 , kadry Mohamed sadek 2 , Heba Farouk Elaarag 3 1 Prof. of meat hygiene , technology and food safety Faculty of Veterinary Medicine, Damanhur university 2 Assistant prof. of bio-chemistry safety Faculty of Veterinary Medicine Damanhur University. 3 Bachelor of veterinary medicine Kafr El Sheikh University Abstract Meat is considered an excellent source of nutrient for human through its contents of protein, mineral and vitamins. In addition, it considered a suitable media for multiplication of spoilage microorganisms and food borne pathogens. Papaya contains powerful antioxidant as vitamin C, E and flavonoids; the B vitamins, folate and pantothenic acid. Papaya and its extracts have antibacterial activity against Staphylococcus aureus, Pseudomonas, E-coli, Proteus mirabilis, Klebsiella pneumonia and Salmonella typhi. The aim of this work was to replace the use of chemical preservatives by a natural preservative. We studied the possibility of the use of papaya seeds, seeds and leaves extract in preservation of raw minced meat through evaluation of antibacterial and antioxidant capacities. The work was done by dividing meat into 4 groups; one as a control group and the other three groups each divided into three subgroups to test the effect of papaya seeds, papaya seeds and leaves extracts with (1%, 2% and 3%). Each concentration was added to the meat groups and preserved at 3:5 o C. A sample from each group was taken every day from day zero till signs of spoilage were appeared. We also compared their effect on Staph aureus and E. coli and on meat quality through measuring total antioxidant capacity and lipid peroxidation. The results revealed that papaya leaves extract (3%) gave the best result and the longest period of preservation with lowest bacterial count followed by seeds extract at concentration 3%. Dried powdered papaya seeds without extraction nearly had no effect. In conclusion, preservation of meat with leaves extract 3% is preferred than other tested methods of preservation. 1- Introduction Meat is considered an excellent source of nutrients for human through its contents of protein, mineral and vitamins . also a suitable media for multiplication of spoilage microorganisms and food born pathogen through its content of moisture, nitrogenous compounds, mineral, growth factors, fermentable carbohydrates (glycogen) and favourable pH (Aymerich et al., 2008).
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International Food Safety Conference th5
Damanhour University
October 2018 th, 13Saturday
337
Effect of Carica Papaya seeds, seeds and leaves extract on Staphylococcus aureus and
Escherichia coli in minced meat and on meat quality
Ebeed Abd Elatty Saleh 1, kadry Mohamed sadek 2, Heba Farouk Elaarag 3
1 Prof. of meat hygiene , technology and food safety Faculty of Veterinary Medicine,
Damanhur university
2 Assistant prof. of bio-chemistry safety Faculty of Veterinary Medicine Damanhur
University.
3 Bachelor of veterinary medicine Kafr El Sheikh University
Abstract
Meat is considered an excellent source of nutrient for human through its contents of protein,
mineral and vitamins. In addition, it considered a suitable media for multiplication of
spoilage microorganisms and food borne pathogens. Papaya contains powerful antioxidant as
vitamin C, E and flavonoids; the B vitamins, folate and pantothenic acid. Papaya and its
extracts have antibacterial activity against Staphylococcus aureus, Pseudomonas, E-coli,
Proteus mirabilis, Klebsiella pneumonia and Salmonella typhi. The aim of this work was to
replace the use of chemical preservatives by a natural preservative. We studied the possibility
of the use of papaya seeds, seeds and leaves extract in preservation of raw minced meat
through evaluation of antibacterial and antioxidant capacities. The work was done by
dividing meat into 4 groups; one as a control group and the other three groups each divided
into three subgroups to test the effect of papaya seeds, papaya seeds and leaves extracts with
(1%, 2% and 3%). Each concentration was added to the meat groups and preserved at 3:5oC.
A sample from each group was taken every day from day zero till signs of spoilage were
appeared. We also compared their effect on Staph aureus and E. coli and on meat quality
through measuring total antioxidant capacity and lipid peroxidation. The results revealed that
papaya leaves extract (3%) gave the best result and the longest period of preservation with
lowest bacterial count followed by seeds extract at concentration 3%. Dried powdered papaya
seeds without extraction nearly had no effect. In conclusion, preservation of meat with leaves
extract 3% is preferred than other tested methods of preservation.
1- Introduction
Meat is considered an excellent source of nutrients for human through its contents of
protein, mineral and vitamins . also a suitable media for multiplication of spoilage
microorganisms and food born pathogen through its content of moisture, nitrogenous
compounds, mineral, growth factors, fermentable carbohydrates (glycogen) and favourable
pH (Aymerich et al., 2008).
International Food Safety Conference th5
Damanhour University
October 2018 th, 13Saturday
338
Contamination of fresh meat starts from live stock rearing environment,
transportation to slaughter house, improper processing practices, usage of dirty contaminated
tools, unhygienic slaughter house environment, ill workers and contamination during
transportation from slaughter house to shops which result to spread of microorganisms from
gastrointestinal track to muscle till improper handling in retail shops (Afshin et al., 2011).
Food born diseases and microbial spoilage result from failure to control
microorganisms at one or more stages from production to consumption (Normann et al.,
2005). These food born infections result in economic and health losses (Adeyemo, 2002).
Staphylococcus aureus is one of most important food born pathogen as it produces
thermostable enterotoxin (Tang and Stratton, 2010), this toxin cause vomiting and diarrhea
(Hennekinne et al., 2012) .
E. coli is the most dominant aerobic commensal flora in gastrointestinal track of
animal and human, it is considered an indicator as a source of fecal contamination if presents
in meat and some serotypes of E.coli lead to death of young children and cause cholera like
syndrome (Gyles, 2007).
Structure of meat is a complex physical structure and chemical composition that is
very susceptible to oxidation . Appearance, color, texture and flavor are the main sensory
quality affect consumer acceptance of meat, and lipid peroxidation is the primary cause of
these quality deteriorations in meat (wood et al., 2008). Lipid peroxidation primarily occurs
through a free radical chain reaction, and oxygen is the most important factor on the
development of lipid peroxidation in meat (Decker, 1992)
Carica papaya belonges to the family of Caricacae, and several species of this family
have active compounds which are valuable in the treatment and control of many diseases.
(Alabi et al., 2012). Papaya contains powerful antioxidant as vitamin C and flavonoids; the B
vitamins, folate and pantothenic acid; and minerals , potassium and magnesium; which
protect against colon cancer and keep function of cardiovascular system (Arvind et al.,
2013). Papaya also has anti-helminthic and anti- amoebic (Arvind et al.,2013) and anti-
inflammatory activity (Oladunmoye and Osho, 2007). Papaya has properties to act as
analgesic, cardiotonic, cholagogue, hypotensive, digestive, laxative and
stomachic(Anibijuwon and Udeze, 2009).
Papaya seeds protect from kidney failure through their effect on kidney toxins and also
have antimicrobial activity against Trichomonas vaginalis trophozoites so it can be used for
urogenital disorder with care to avoid toxicity and chewing of seedes used for treatment of
intestinal worm (Peter et al.,2008). Also Leaves of Papaya tea or its extract have renown to
be used as anti-tumer agent (Baskaran et al.,2012). Papaya seeds and its extract and papaw
leaf extract have antibacterial activity against Staphylococcus aureus , Pseudomonas
aeuginosa, E-coli, Proteus mirabilis, Klebsiella pneumonia and Salmonella typhi (Peter et
International Food Safety Conference th5
Damanhour University
October 2018 th, 13Saturday
339
al., 2014). Papaya seeds contain active ingredients which have antibacterial and antioxidant
effects (Peter et al.,2008). Also leaves contain phenolic compounds which have powerful
antioxidant effects and antimicrobial effects (Romasiet al.,2011). The digestive properties of
papaya is related to papain which treat dyspepsia. Papain also has clotting properties to milk
(rennet) and it is useful for tenderization of meat (Udohet al., 2005).
Therefore, the aim of this work was planned for studying the effect of carica papaya
seeds, leaves and seeds extract on count of staphylococcus aureus and E-coli and on total
antioxidant capacity and lipid peroxidation in minced meat.
2- Materials and Methods
2.1.Collection of samples :
Sample of 10 kg of minced meat collected from different butcher shops in kafr El Zayat city.
Carica Papaya fruit and leaves collected from trees in kafr El Zayat and surrounding villages
2.2. Preparation of leaves and seeds extracts :
2.2.1. Preparation of hot aqueous seed extract : (Oyagade et al., 1999).
2.2.2. Preparation of aqueous leaves extract : (Oyagade et al., 1999)
2.3. Dividing of minced meat into parts and addition of of papaya extracts and seeds :
10 Kg of minced meat were divided into 100 part which divided into 4 groups :
Group 1 : 10 parts consist of 1 kg used as control sample
Group 2 : 30 parts consist of 3 Kg used for addition of leaves extract
This group divided into 3 sub groups with 3 concentrations of leaves extract each one
consists of 10 part of 1 kg weight
Sub-group 1 : add concentration 1% of leaves extract (10 gm extract / 1 kg meat)
Sub-group 2 : add concentration 2% of leaves extract ( 20 gm extract / 1 kg meat)
Sub group 3 : add concentration 3 % of leaves extract ( 30 gm extract / 1 kg meat )
Group 3 : 30 parts consists of 3 kg for addition of seeds extract and divided into 3 sub-
groups with different concentration of extract as described before in group 2
Group 4 : also consists of 3o part of 3 kg for addition of seeds and also divided into 3 sub
groups as in group 2 and 3.
preservation by refrigeration at temperature 3:5oC and take every day sample from each
group started from 1st day of addition of herbs until spoilage signs begin to appear on
samples. And make counting to staph. aureus and E.coli and compare count in all groups .
And also studying the antioxidant effect of papaya extract by :
2.4.determination of total antioxidant capacity and lipid peroxidation.
Preparation of samples (ISO,2007)
International Food Safety Conference th5
Damanhour University
October 2018 th, 13Saturday
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2.5.Microbiological examination :
A) Determination of Staphylococcus count (ICMSF, 1996)
B) Identification of suspected Staphylococcus aureus
Morphological identification
1- Gram’s stain (cruickshank et al.,1978)
2- Motility test (ICMSF,1996)
Biochemical identification of S.aureus (ISO,2007)
1- Coagulase test (tube technique)(APHA,2001)
2- Oxidation-fermentation of glucose(O-F test)
3- Catalase test (slide technique)(MACFADDIN, 1976)
4- Mannitol fermentation test (Baily and Scott,1978)
5- Urease test
6- Thermo-stable nuclease production
7- Detection of haemolysis (Baily and Scott,1978)
C)Isolation , identification and counting of E coli :
Morphological examination
1- Staining: (Cruickshank et al.,1975)
2- Motility test (ICMSF,1978)
biochemical identification (FDA,1998)
1- Indole production test
2- Methyl red test
3- Voges – praskauer test
4- Citrate utilization test
2.6.Determination of lipid peroxidation and total antioxidant capacity (Satoh, 1978)
1-Determination of lipid peroxidation
2-Determination of total antioxidant capacity
3- Results and Discusion:
Meat and meat products are considered the major vehicle of most food borne microorganism.
Contamination of meat and meat products starts from abattoir, utensils, environments,
transportation, food handlers and manufacturing. Unhygienic practices during meat handling
International Food Safety Conference th5
Damanhour University
October 2018 th, 13Saturday
341
result to growth and multiplication of spoilage and pathogenic microorganisms which lead to
public health hazards. ( Isis et al., 2009).
Staphylococcal count:
Staphylococci commonly present on skin and upper respiratory tract of human and
animals so, staphylococci can easily contaminate meat and meat products if the control of
hazards is not occur during any step from farm to fork and occur contamination by bacterial
or its toxins lead to gastroenteritis when occur consumption of food.
In our study we found that the mean value of count of Staphylococcus aureus in minced meat
was 8.8×103 CFU/g. as shown in table (1). This result similar to the range reported by (Reda,
2010) , (Marwa, 2017) and (Shimaa, 2016)
According Egyptian standard (E.O.S.Q.C, 2005) the permissible staphylococcal count is 102
CFU/g.
Table (1) The effect of different concentrations of herbs on count of Staph. aureus
(CFU/g) in minced meat during storage under refrigeration.
Day 1 Day 2 Day 3 Day 4 Day 5 Day 6 Day 7 Day 8
Control 8.8×103 12.3×103 4.8×104 6.0×104 Decomposed