Journal of Environmental Treatment Techniques 2015, Volume 3, Issue 4, Pages: 201-203

201

Effect of Residual Antibiotics in Snacks against Escherichia coli

and Staphylococcus aureus

Lovet T. Kigigha, Sylvester Chibueze Izah* and Phillip E. Prebo

Department of Biological Sciences, Faculty of Science, Niger Delta University, Wilberforce Island, Bayelsa state, Nigeria

Received: 15/12/2015 Accepted: 27/12/2015 Published: 30/12/2015

Abstract

This study assessed the effect of ethanolic, hot water and cold water extracts of processed ready-to-eat snacks containing

protenous substances against Escherichia coli and Staphylococcus aureus. The snacks (shawama, Scotch egg and meat pie)

were purchased from fast food hawkers in Yenagoa metropolis, Bayelsa state, Nigeria. Disk-diffusion method was used for the

sensitivity testing of the various extracts and 1% Ampiclox was used for the comparison of the antibacterial effects. Results

showed that ethanolic extracts have superior effect compared to cold and hot water. For hot water treatment, the E. coli zone

of inhibition for scotch egg, shawama and meat pie were 8.38, 12.57 and 10.48mm respectively. For cold water extracts, these

were 10.48, 13.62 and 8.38mm respectively. For the ethanolic extracts, the zone of inhibition for the snacks was 12.57mm

(Scotch egg), 14.67mm (shawama) and 13.62mm (meat pie). For S. aureus, the corresponding zones of inhibition for the

snacks i.e scotch egg, shawama and meat pie were 10.48, 14.67 and 8.38mm respectively (for hot water extract), 10.48, 15.71

and 12.57mm respectively (for ethanolic extract) and 10.48, 12.57 and 10.48mm respectively (for cold water extract). Ethanolic

extract had higher zones of inhibition on both isolates. Also the E. coli had superior zone of inhibition compared to 1%

Ampiclox than S. aureus. Analysis of variance showed that there was significant difference (P<0.05) among the test organisms

used in this study based on the treatments. This study showed that residual antibiotics used for protenous sources (e.g egg, meat

etc) were transferred to their final products widely consumed by several people as fast foods, which are becoming more popular

with students and the working class who spent long hours from home.

Keywords: Food, Protein, Snacks, Escherichia coli, Staphylococcus aureus

1 Introduction

Food is a composite matrix that contains nutrients to

enhance growth [1]. Food substances include

carbohydrate, protein, lipids and water etc. Ready-to-eat

foods or snacks are foods that do not require further

processing prior to consumption. Most of these ready to

eat foods are vended along highways, streets, markets and

some public places including schools, hospitals, Motor

Parks, etc. Some common ready to eat products include

tomato, cabbage, ginger drink, lettuce, carrot, kunu zaki

drink, akara, fried groundnut, rice and beans, fired fish,

bread, zobo drink [2], and snacks such as scotch egg,

shawama, meat pie etc. In developing nation like Nigeria,

the consumption of snacks is high. This could be due to

high rate of unemployment and failed family and

community values [3]. Like fruits most ready-to-eat food

are rich in nutrients, micronutrients, vitamins and fibre.

Like most ready to eat food products, Scotch egg,

shawama, meat pie are sold in public places including

markets, fast food, stores, restaurants etc. During their

production, several natural and protenous ingredients are

used. For instance, poultry egg is used for the production

of Scotch egg. The poultry industry in which poultry-meat

is considered healthier than red meat is a vital source of

animal proteins [4]. In the poultry industry antibiotics are

used for the rearing of the birds due to several microbial

diseases that infects birds. As such antibiotics and growth

promoters are commonly used by poultry farmers for the

control infections [4]. Also, during the processing of cotch

egg, shawama, meat pie, other natural products with

Corresponding author: Sylvester Chibueze Izah, Department of Biological Sciences, Faculty of Science, Niger Delta

University, Wilberforce Island, Bayelsa state, Nigeria, E-mail: chivestizah@gmail.com; Tel: +234 703 0192 466.

antimicrobial activities such as pepper are added at varying

concentrations.

E. coli is a typical example of the family

Enterobacteriaceae which are associated with

gastroenteritis. The Staphylococcus aureus on the other

hand represents the superficial agents of infection. The

study therefore meant to test antibacterial inhibition

caused by residual antibiotics in some common snacks and

comparing their inhibition with that of 1% broad spectrum

antibiotics (Ampiclox).

2 Materials and Methods

2.1 Source of samples

Triplicate samples of the snacks (Scotch egg, meat pie

and shawama) were bought from fast food hawker in

Yenagoa metroplolis, Bayelsa State, Nigeria and

transported to the laboratory. Analysis was carried out <4

hours after purchase.

2.2 Preparation of Sample Extracts

In the laboratory, the various samples (Scotch egg, meat

pie and shawama) were separately macerated in a sterile

mortar and pestle. Then 10g each of the macerated samples

were soaked in 100ml of solvents (ethanol, hot water and

cold water) separately, which was vigorously shaked

intermittently for 24 hours. Thereafter, the samples was

filtered with muslin cloth followed by Whiteman No. 1

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ISSN: 2309-1185

Journal of Environmental Treatment Techniques 2015, Volume 3, Issue 4, Pages: 201-203

202

filter paper. All extracts were persevered at 5ºC in the

refrigerator prior to the sensitivity testing.

2.3 Source of bacterial isolates

Test pathogenic bacteria isolates i.e S. aureus and E.

coli used for the study were obtained from Glory land

Clinic, Yenegoa, Bayelsa State of Nigeria. The purity of

the isolates was tested by subculturing and they were

preserved in nutrient agar slants at 5

C in the refrigerator

prior to use.

2.4 Impregnation of filter-paper discs with the extracts

and Antibacterial assessment

The disk-diffusion technique previously described by

Kigigha and Atuzie [5], Kigigha and Charlie [6], Kigigha

and Onyema [7] were used to test the antibacterial activity

of the different extracts using 10 mm discs. Whatman No.

1 filter paper was used to prepare the antibiotic discs using

cock borer. The solution containing the extracts (0.1ml

each) were impregnated on the disc using sterile pipette

and air- dried. 1% Ampiclox solution (which served for

comparison) was also impregnated on filter-paper discs.

Flame-sterilized and cooled pair of forceps was used for

the transfer of the discs separately. The different

treatments were placed separately on nutrient agar plates

with lawn spread growth of the test isolates. The filter

paper impregnated with 1% Ampiclox solution was also

placed separately on the lawn spread test isolates. The

discs were well spaced out on the agar plates to avoid

overlapping of the zones of inhibition. The plates were

incubated at 37°C for 72 hours to observe and measure the

zones of inhibition.

2.5 Statistical Analysis

Data was analyzed using Zigma-Stat 32 Statistical

package. Where normality and homescedasticity were

satisfied, data from treatment were analyzed using one-

way analysis of variance (ANOVA). Comparison for

treatments was analyzed using Tukey tests at P= 0.05.

3 Results and Discussion

Figure 1 presents the antibacterial effects of the snacks

on some pathogenic microbes which was observed for hot

water scotch egg extracts were 8.38 and 10.48mm for

E.coli and S. aureus respectively; for hot water shawama

extracts were 12.57 and 14.67mm for E.coli and S. aureus

respectively and for hot water meat pie extracts were 10.48

and 8.38mm for E.coli and S. aureus respectively. The

various ethanolic extracts of the snacks were 12.57 and

10.48mm for E.coli and S. aureus respectively (scotch

egg), 14.67 and 15.71mm for E.coli and S. aureus

respectively (shawama) and 13.62 and 12.57mm for E.coli

and S. aureus respectively (meat pie). Also the cold water

extracts for the various snacks was 10.48 and 13.62mm for

E.coli and S. aureus respectively (scotch egg), 10.48 and

12.57mm for E.coli and S. aureus respectively (shawama)

and 8.38 and 10.48mm for E.coli and S. aureus

respectively (meat pie).

Fig 1: Zone of Inhibition for some fully processed snacks (shawama, meat pie and Scotch egg). (Ec= E.coli, Sa = S. aureus, SE = Scotch egg, SW= Shawama,

MP=Meat pie, Etol =Ethanol estracts, CW = Cool water extracts ; HW= Hot water extracts); n=3

The zone of inhibition for the 1% Ampiclox were

19.90 7.33mm for E.coli and S. aureus respectively.

Basically there was significance difference (p<0.05)

among the various treatment in both bacterial isolates. This

could be due to the various composition and level of

incorporation of spices/preservatives during their

processing. Again, the level of heating the products are

subjected to could have also contributed to the

antibacterial efficacy of the various snacks under study.

The results fluctuates for instance, S. aureus was higher in

scotch egg and shawama samples and lower for meat pie

(hot water extracts), E. coli have superior zone of

inhibition for scotch egg and meat pie but lower for

shawama) and cool water extract were consistent showing

Antimicrobial activity of Some Protenous Snacks

EcHW/SE

SaHW/SE

EcHW/SW

SaHW/SW

EcHW/MP

SaHW/MP

EcEtol/SE

SaEtol/SE

EcEtol/SW

SaEtol/SW

EcEtol/MP

SaEtol/MP

EcCW/SE

SaCW/SE

EcCW/SW

SaCW/SW

EcCW/MP

SaCW/MP

Ec AMPI

Sa AMPI

Treatment of Protenous Snack

Zones of Inhibition (mm2)

Journal of Environmental Treatment Techniques 2015, Volume 3, Issue 4, Pages: 201-203

203

that S. aureus has superior zone of inhibition than E. coli.

However, Ampiclox has lower and highest zone of

inhibition for S. aureus and E. coli respectively. The trend

of ampiclox having lesser effect in S. aureus when

compared to E.coli in agreement with the findings of

several authors on different substrates [5 – 7]. On the

overall, ethanol had higher zone of inhibition when

compared to other extracts (hot water and cold water). The

trend in this study was in conformity with findings of

Masih et al. [8], Ere et al. [9], Akintobi et al. [10] on

antibacterial effects of different plant species on some

microbes including the ones used in this study. Opoku and

Akoto [11] has attributed the presence of insoluble active

compound found in cold water and or denaturation of the

active constituents by the hot water extraction process as

potential reason why they have lower zone of inhibition.

The antibacterial potentials in the snacks (shawama,

scotch egg and meat pie) could stem from the fact that

livestock is an ingredient in the production of this snacks.

Poultry products including its meat (used for the

processing of the meat pie and shawama depending on

choice), egg (used for the processing of the scotch egg)

could be source of the antibacterial agent in the products.

Sometimes, the preservative or spice added to the food

products could also be potential antibiotics in the snacks.

4 Conclusion

This study evaluated the antimicrobial effects of some

fully processed snacks such as scotch egg, shawama and

meat pie prepared with poultry products on bacterial

isolates. The study found that the snacks have antibacterial

effects against common enteric and superficial etiologic

agents thus E.coli and S. aureus respectively. This is an

indication that drug-resistant in normal human flora could

be elated through eating of snacks. This also highlighted

the need to cure farm animals and their products of residual

antibiotics before they are sold to public. This could be

achieved through the use of prebiotics and probiotics and

botanicals in the rearing of livestocks. Also it appears to

be no legislation in Nigeria in this respect.

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