ABSTRACT
Essential oil produced from turmeric and Bacteriocin produced from Lactobacillus plantarum were used to control Salmonella species found in dairy products. Six dairy products inculding milk powdered,cheese,icecream,yoghourt, tin milk, fresh milk were screenced for Salmonella species, only three isolates were identified which inculde Salmonella typhi, Salmonella enteric, Salmonella chloeraceau. Results showed the presene of Salmonella species in three products yoghourt, fresh milk and milk powdered. Lactobacillus was isolated from “ogi”(fermented maize starch) and it’s yield of bacteriocin was 0.36% while essential oil yield from turmeric was 4.94%. The highest zone of inhibition was 24.33mm were recorded for bacteriocin activity aganist Salmonella species while the highest zone of inhibition for essential oil showed antibacteria activity with zone of inhibition in diameter of 14.33mm aganist the isolates .The test of biocontrol agents more also are found to be active at low concentraction of 15mg/ml and 21mg/ml for bacteriocin and essential respectively. Comparatively the bacteriocin extract was found to have higher activity than the essential oil extracts and should be incoperated in strater culture in production of dairy products.
TABLE OF CONTENTS
Title page i
Certification ii
Acknowledgments iii
Dedication iv
Table of Contents v
List of tables vi
Abstract vii
CHAPTER ONE
1.0 Introduction
1.1 Introduction 1
1.2 Aim and Objectives 2
CHAPTER TWO
2.0 Literatue Review 4
2.1 Latic acid bacteria 4
2.2 Latic acid bacteria and bacteriocin 6
2.3 Bacteriocin 7
2.4. Classification of bacteriocin 8
2.4.1 Class 1 (Lantibiotics) 8
2.4.2 Class 11 (Small heat stable peptides) 9
2.4.3 Class 111(Large heat labile bacteria) 9
2.4.4 Class iv 9
2.5.0 Essential oil from turmeric 10
2.5.1 Scientific classification of turmeric 10
2.5.2 origin and uses of turmeric 11
CHAPTER THREE
3.0 Materials And Methods 15
3.1 Materials 15
3.2 Methods 15
3.2.1 Sample Collection 15
3.2.2 Media Prearation 15
3.3.0 Isolation Of Salmonella From Dairy Products Determination of Micriobiral
Load (cfu/g) 15
3.3.1 Confirmation of Salmonella 16
3.3.2 Microscopic Examination/Gram Staining 17
3.4.0 Sources of Lactic Acid Bacteria 17
3.5.0 Biochemical Tests 18
3.5.1 Indole Test 18
3.5.2 Cirtrate Utilization Test 18
3.5.3 Nitrate Reduction Test 18
3.5.4 Catalase Test 19
3.5.5 Cogulase Test 19
3.5.6 Oxidase Test 19
3.5.7 Sugar Fermentaion Tests 19
3.7.0 occurance of Salmonella in Dairy Products 20
3.8.0 Extracton Of Biocontorl Extracts (Bacteriocin From Lactobacillus plantarum and Essental Oil From Turmeric/Determination Yield 20
3.8.1 Extracion of Essential Oil from Turmeric/Determination Yield 20
3.8.2 Extraction of Bacteriocin /Detremination Yield 21
3.9 Antibacterial Activity Test 22
3.10 Antibiotics Senstivity Test 23
3.11 Minimum Inhibtion Concentraction (MIC) 23
3.12 Statistical Method 23
CHAPTER FOUR
4.0 Results 24
CHAPTER FIVE
5.0 Discussion, Conclusion And Recommendation 29
5.1 Discussion 29
5.2. Conclusion 30
5.3 Recommendation 31
References 32
Appendix
List of Tables
Tables Title Pages
1. Occurance of Salmonella Species in Dairy Products 25
2. Precentage Yield Of Esseential oil And Baceteriocin 26
3. Zone of Inhibition of Bacteriocin and Essential Oil Aganist 27
Salomonella Species in Diameter (mm)
4. Minimum Inhibition Concentration of Essential Oil and 28
Bacteriocin of Salmonella Species.
CHAPTER ONE
1.0 INTRODUCTION
Inspite of morden improvement in food production techiques, food safety is still an improtant aspect of public health .It has been estimated that as many as 30% of people in industrised countries has most occuring bacterial food borne disease each year (WHO, 2002) defined food disease as disease infection or toxification caused by the consumption of containmated food or water. Salmonellosis was the most occuring bacterial food borne disease with more than 325,000 cases per year (Baeulmer et al, 2005) .Food borne outbreak of Salmonella assoicated with consumption of contaminated dairy products and other ready-to- eat products has been reported to occur with increased frequancy Doyle (1992), Baeulmer et al 2005, Bredhoit et al, 2005) also Staphylococus anerus, Eschericha coli cause food borne infection. Salmonella belongs to the family of enterobacterioaceae which inculdes the genera of Salmonella ,Proteus, Klesbsiella,Shigella, Enterococus, Eschericha coli, there up to 200 serotypes and 70 sub-species of Salmonella (Jawetz et al, 2005).
According to cheesbrough 2006,Salmonella are gram negtive bacteria with rods,areobes, facuitves they grow between the temperature of 15-450C. They are motile withe pertriochas flagella , they ferment maltose, manitole .They produce hydorgen suphides, non –sporing ,non-capsulated ,biochemical test of Salmonella is urease, indole negtive and cirtates postive. The natural habitat is intestinal tract of human’s and animals ,the main infective dose is 105-108(CDCP,1993). Some Enteric organism are part of normal microbiota and incidently cause disease, but Salmonella-Shigella are pathogenic to humans. salmonella grows in simple media and selective media such as Salmonella-Shigella ager (SSA). They survive freezing and can stay for longer period, they are isolated from food and other poducts such as dairy products there symptoms inculdes vomitting ,fever,diarrohea and abdomial pains after ingestion of the infected foods between 12-36hours (Baeulmer et al 2005). Colony morophololgy , identification of Salmonella on SSA are in cluster and has black pigment.
According to Reidtke et al (1993) biocontrol agents refers to the suppression, reduction or inactivation of food borne pathogen bacteria by other bacteria or micobiral agents. Progress in research and advanced in techonology has led to various practical ways for microbiral control of food borne . Biocontrol agents have anti microbiral activites aganist Staphylococus auerus, Escheriha coli, Pseudomonas and other pathogenic oraganism (Parish et al 2003, Liao et al 2010) .Biocontrol are also known as biological control or bioactive microbes may inhibit or suppress food borne pathogen. Latic acid bacteria (LAB) can be used as biocontrol agent or its extract like Bacteriocin. LAB are group bacteria gram negtive bacteria that can be used as biopreservative ,bicontrol .They are used to prolong life shelf of food by producing a number of organic compounds that that are antagonistic to other microorganism (Brink et al, 1994). Among these compounds proteinaeous bacteriocin has grained much attention especially regarding to much reserach focused on utilization of bacteriocin as Novel food biopreservatives ,also the interest in using them as biocontrol of bacteria in human because of the probiotics potenial of these bacteria is also vastly investiagated ( Redtke et al, 1993, Onyeagba et al 2004 , Diez-Gonzaales , 2007). Alot of Lactobacillus has been noted to have nutitional benfits, imprived lactose utilization have anti-cholesterol and carcinogenic activites and protection agenist other disease. Bacteriocin has received attention for use as natural or Biopreservatives in food in recent years (Savadogo et al, 2004).
Accoring to Dimick et al (1947) using solvent extraction method to extract curde bacteriocin from Lacto-bacillus Plantarum. In recent year use of essential oil (EOs) as biocontrol agent has recevied attention both in pharamcetical, comestic,agrcultural and food indusrty because of the biological activity, notably antibacterial, antifungal, activity and efficeney of EOs to be used as antimicrobiral agents in food system depends on the factors as temperaure, PH, apperance, load of microbiral flora and favourable evironment. In recent years, serveral research have shown that many spices and herbs who exert antimicrobial activity due to their essential oil fraction can be used in food system for inhibiting the growth of pathogenic microorganism this way ensuring the microbiological saftey of food products (Bankkali et al, 2008, Burt, 2004,Yardugul et al, 2002), and that antimicrobiral effects of EOs exhibit strctural or functional damges of membrane of pathogenic bacteria by various antimicrobiral mechanism (Cotter et al 2000, Burt 2007) the plant EOs is gotten from Turmeric (Curcuma longa) the EOs is extracted by solvent extraction method according to (Prasad et al 2001). Turmeric is a pereninal plant that is cultivated throughout tropices of Asia,india and chian,the plant grows at the height of 0.9-1.5m and bears large oblong leaves and funnel-shaped, dull yellow flowers. recently ,The EOs strated to used because of their antimicrobiral activity in food package,to edible films provide incoprating of compounds into edible films provides a noval way to improve the safety and shelf life of ready-to-eat food (Diez-Gonzale et al, 2007, cutter 2000).
1.2 Aim And Objectives
To evaluate inhibitory activities of bacteriocin and essential oil aganist salmonella species on dairy products.
i. To determine the antibacterial activites and Minimum Inhibitory Concentraction of bacteriocin and essential oil
ii. Isolation and identification of Salmonella from different dairy products .
iii. Extraction of biocontrol agents (bacteriocin from Lactobacillus planatrum and essential oil from turmeric).
iv. To determine the effects of these bicontorl agents aganist Salmonella in dairy products to reduce foodborne infection.
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