SHEIKH TECHNICAL VETERINARY SCHOOL (STVS)

Assessment of E. coli Contamination of Meat

(Case study: Galkaio Meat Market)

A mini thesis submitted to sheikh technical veterinary school as a partial requirement for the award of Diploma in livestock product development and entrepreneurship (DLDE)

Prepared by

Ali Mohamed Ali Iye

Supervisor/Advisor:

Dr. Abdullahi Ali Herzi

Jul 2012 1

DECLARATION

The work presented here is my own and has not been submitted to any other university for the award of a Diploma.

Signed

Ali Mohamed Ali Iye

Date

This thesis is submitted with the approval of my supervisors

Signed

Dr. Abdullahi Ali Herzi

Date

ACKNOWLEDGEMENTS

I wish to thank the following people for their kind assistance in both the Research and preparation of this work: Dr.Abdullahi Ali Herzi: My sincere appreciation for the supervision and assistance you have given me; for the trust and confidence you showed in my abilities and for your truly caring nature which inspired me to do my very best. Mr. Abdinasir Ali Mohamed: Your support has given me a good head start towards a challenging but exciting career. Your constructive criticism helped me challenge myself and to keep learning and growing. Mr. Asad Mohamed Hirsi: For assistance with the laboratory analysis. Without your input my work would have remained untested and therefore had incomplete meaning All the staff of Galkaio veterinary laboratory: Thank you to all of you. Staff from sheikh technical veterinary school and all people whose names I have not been able to mention here, but who have made invaluable contributions to my work directly and indirectly; my sincere appreciation to all of you.

DEDICATION

I would like to dedicate this work to my late parents Idiil Mohamed Farah And Mohamed Ali Iye, for their unconditional love, for Teaching and showing us that in life someone has to work hard in order to earn a living (May their souls rest in peace). To my brothers and sisters all my nieces for their moral support. Last but not least, I would like to thank my colleagues with whom we have Travelled this journey through thick and thin, the Diploma in livestock product development and entrepreneurship (DLDE) graduate group.

List of Abbreviations and symbols

BPW BGBB CDC FAO GMP HACCP LTB MPN STVS

Buffered Peptone Water Brillian Green lactose Bile Broth Center for Disease Control and Prevention Food Agricultural Organization Good Manufacturing Practice Hazard Analysis Critical Control Point Lauryl-sulfate Tryptose Broth Most Probable Number Sheikh Technical Veterinary School

Table of Contents DECLARATION............................................................................................................................2 ACKNOWLEDGEMENTS...........................................................................................................3 DEDICATION...........................................................................................................................4 List of Abbreviations and symbols.............................................................................................5 Table of Contents...........................................................................................................................6 Introduction................................................................................................................................7 1.2.1 Escherichia coli ................................................................................................................8 1.2.2 Epidemiology....................................................................................................................8 1.2.3 Susceptible populations:..................................................................................................10 1.2.4. PATHOGEN-FOOD COMMODITY COMBINATION(S) OF CONCERN................10 Escherichia coli........................................................................................................................10 1.2.5 Pathogen of concern........................................................................................................10 1.2.6 Sources and Modes of Transmission...............................................................................11 1.2.7 Symptoms of E. coli infection:........................................................................................12 1.2.8 Diagnosis: Food quality/safety indicator tests ................................................................13 Aerobic Plate Count ................................................................................................................13 1.2.9 Public health significance:...............................................................................................13 1.2.9.1 Sources of E. COLI O157:H7 contamination..............................................................14 1.2.9.2 Person-to-Person transmission of E. coli.....................................................................14 1.2.9.3 Prevention of E. COLI O157:H7 contamination through use of colicinogenic E. COLI strains:......................................................................................................................................14 1.2.9.4 Prevention of E.COLI O157:H7 contamination through application of HACCP........14 1.3.1 Aims................................................................................................................................16 1.3.2 Objectives........................................................................................................................16 Chapter two: Material and methods.............................................................................................17 2.1 Study site................................................................................................................................17 2.2 Material and Methods.........................................................................................................18 ..............................................................................................................................................19 BGBB (confirmation media) LTB (selective enrichment media)..........19 2.3 E. coli confirmation ...........................................................................................................19 Chapter three: Result and Discussion...........................................................................................20 3.1 Result ................................................................................................................................20 4.1 Conclusion:.........................................................................................................................24 4.2 Recommendations:.............................................................................................................24 List of References.........................................................................................................................25 Annexes .......................................................................................................................................27 Questioner format:....................................................................................................................27

Chapter One: Introduction and Literature review Introduction Food security is a complex issue, where animal proteins such as meats, meat products, fish and fishery products are generally regarded as high risk commodity in respect of pathogen contents, natural toxins and other possible contaminants and adulterants (Yousuf et. al., 2008). Food borne infections and illnesses is a major international health problem with consequent economic reduction. It is a major cause of illness and death worldwide (Adak et. al., 2005). Recognizing this, the World Health Organization (WHO) developed its Global Strategy for Food Safety (Adak et al., 2005). There are four major pathogens that have frequently been associated with meat and meat products including Escherichia. coli, Salmonella spp., Campylobacter spp., Listeria monocytogenes (Adak et al., 2005 ). These organisms have been linked to a number of cases of human illness. One of the most significant food-borne pathogens that has gained increased attention in recent years is E. coli O157:H7. Typical illness as a result of an E. coli O157:H7 infection can be life threatening, and susceptible individuals show a range of symptoms including hemolytic colitis, hemolytic-uremic syndrome, and thrombotic thrombocytopenic purpura. Domestic and wild animals are the sources of E. coli O157, but ruminants are regarded as the main natural reservoirs. Sporadic cases and outbreaks of human diseases caused by E. coli O157 have been linked to ground beef, raw milk, meat and dairy products, vegetables, unpasteurized fruit juices and water. Infections can also be acquired by direct contact with animals and by person-to-person spread. Currently, there is limited information regarding on meat contamination of E. coli O157:H7 in Somalia. Therefore, this study was conducted to determine the contamination level of E. coli, from Galkaio Meat Market.

1.2 Literature Review 1.2.1 Escherichia coli In contrast to either coliforms or fecal coliforms, E. coli has a taxonomic basis (Hitchins, et al., 1992). E. coli is a member of the family Enterobacteriaceae (Ewing, 1986), which consists of many genera, including known pathogens such as Salmonella, Shigella, and Yersinia. Although most strains of E. coli are not regarded as pathogens, some can be opportunistic pathogens that cause infections in immuno-compromised hosts (Gassama, et al., 2001). In addition, there are pathogenic strains of E. coli that when ingested, cause gastrointestinal illness in healthy humans (e.g., E. coli O157:H7). Most pathogenic strains are grouped under the following virotypes: enteropathogenic, enterotoxigenic, enteroinvasive, enteroaggregative, and enterohemorrhagic (Ray, 2004). E. coli was first identified in the intestinal flora of infants by the German pediatrician Theodore Escherich (Escherich, 1885, Bettelheim, 1986), which originally called it as Bacterium coli commune. E. coli is a gram-negative, motile, nonsporulating, rod-shaped, facultative anaerobic bacterium, present in the lower intestinal tract of humans and warm-blooded animals and birds (Ray, 2004) and is the predominant facultative anaerobe in the bowel and part of the essential intestinal flora that maintains the physiology of the healthy host (Conway, 1995).

1.2.2 Epidemiology

In1982,E.coliO157was recognized as ahuman pathogen for the first time and, since then, has been increasingly reported as the cause of illness and outbreaks.E.coliO157:H7 belongs to the group of VTEC of which>200 different serotypes of E.coli have been reported so far, with many of these associated with human disease. The clinical feature of E.coliO157infections include diarrhea, which is often bloody, and may progress to severe hemorrhagic colitis. bout10% of these patients can go on to develop HUS, a potentially life threatening complication characterized by acute renal failure, thrombocytopenia, and hemolytic anemia that is particularly serious in young children and elderly people. 8

Following ingestion of E. coli O157:H7, the human response ranges from asymptomatic infection to death. To cause disease after ingestion, the E. coli O157:H7 must survive acidic conditions within the stomach prior to moving to distal portions of the gastrointestinal tract. Disease due to E. coli O157:H7 occurs primarily in the colon. The incubation period from the time of ingestion to the first symptoms ranges from one to eight days. Asymptomatic shedding of E. coli O157:H7 has been documented (Swerdlow 1997); however, the proportion of exposed individuals who shed E. coli O157:H7 but do not develop symptoms is unknown. Typically the illness begins with abdominal cramps and nonbloody diarrhea that can, but does not necessarily, progress to bloody diarrhea within two to three days (Griffin 1995, Mead et al. 1998). Usually 70% or more of symptomatic patients will develop bloody diarrhea; however, as many as 95% have been observed in other studies (Ostroff et al. 1989; Bell et al. 1994). More severe manifestations of E. coli O157:H7 infection include hemorrhagic colitis (grossly bloody diarrhea), hemolytic uremic syndrome (HUS)10 and occasionally thrombotic thrombocytopenic purpura (TTP). Symptoms of hemorrhagic colitis include severe abdominal cramps followed by grossly bloody diarrhea and edema (swelling), erosion, or hemorrhage of the mucosal lining of the colon (Su and Brandt 1995). Hemorrhagic colitis may be the only manifestation of E. coli O157:H7 infection or it may precede development of HUS. Complications from hemorrhagic colitis associated with E. coli O157:H7 include upper-gastrointestinal bleeding and stroke (Su and Brandt 1995). Roberts et al. (1998, citing Boyce et al. 1995a, Ryan et al. 1986) estimates the mortality rate of those suffering hemorrhagic colitis without progression to HUS to be 1%, although Griffin (personal communication) believes this rate is too high. Approximately 30% to 45% of patients are hospitalized (Ostroff et al. 1989, Le Saux et al. 1993, Bell et al. 1994, Slutsker et al. 1998). Of the 631 cases reported to FoodNet sites in 1999, 39% were hospitalized (CDC 2000b). Treatment for the more serious manifestations of E. coli O157:H7 infection is supportive and the use of antimicrobial agents has been debated (Mead 1998). The incidence of E. coli O157:H7 infection varies by age group, with the highest incidence of reported cases occurring in children. In addition to children, elderly are known to be susceptible to E. coli O157:H7 infection. A report detailing a Scottish outbreak resulting from contaminated beef involving at least 292 confirmed cases of E. coli O157:H7 infection resulted in 151 hospitalizations and 18 deaths; all fatalities were elderly patients (Ahiiied 1997).

1.2.3 Susceptible populations:

The incidence of E. coli O157:H7 infection varies by age group, with the highest incidence of reported cases occurring in children. In addition to children, elderly are known to be susceptible to E. coli O157:H7 infection. While these populations are more susceptible to illness, people of all ages can suffer infection from E. coli O157:H7.

1.2.4. PATHOGEN-FOOD COMMODITY COMBINATION(S) OF CONCERN Escherichia coli E. coli strains that are pathogenic for humans and cause diarrheal illness may be categorized into specific groups based on virulence properties, mechanisms of pathogenicity, and clinical syndromes. These categories include enteropathogenic E. coli (EPEC), enterotoxigenic E. coli (ETEC), enteroinvasive E. coli (EIEC), diffusely-adherent E. coli (DAEC), enteroaggregative E. coli (EaggEC), and enterohemorrhagic E. coli (EHEC). The EHEC group comprises a subset of Shiga toxin-producing E. coli (STEC),8 which include strains of E. coli that cause bloody diarrhea in many infected patients. Shiga toxin-producing E. coli strains produce either or both of two phage-encoded toxins, Shiga toxin 1 (Stx1) and Shiga toxin 2 (Stx2). However, Stx production alone may not be enough to cause illness. Some EHEC strains also contain genes that encode for the ability to attach to and damage intestinal tract cells, causing what is commonly referred to as attaching-and-effacing lesions. E. coli O157:H7 is the single most important EHEC serotype in relation to public health. For a detailed review of the pathogenesis of EHEC and other STEC, interested readers are referred to recent publications by Paton and Paton (1998) and Nataro and Kaper (1998). 1.2.5 Pathogen of concern Enterohemorrhagic Escherichia coli (EHEC) were first identified as human pathogens in 1982, when E. coli strains of a previously uncommon serotype, O157:H7, were implicated in two outbreaks of hemorrhagic colitis (bloody diarrhea) in the United States (U.S.). Since then, 10

outbreaks of this new pathogen have become a serious public health problem throughout many regions of the world (Schlundt 2001; Clarke et al. 2002). The continued occurrence of large outbreaks and an increase in the incidence of reported cases suggests E. coli O157:H7 is an emerging pathogen (Tauxe 1997; Altekruse et al. 1997). Also in the 1990s, EHEC strains of other serogroups such as O26, O103, O111, and O145 were increasingly linked to human illness. This is also suggested by the WHO, which reported that O26, O103, O111 and O145 are the most important nono-O157 serogroups (WHO 1998). Three outbreaks in the U.S. have been ascribed to non-O157 EHEC: a family outbreak of E. coli O111 with a case of HUS, a milkassociated episode of E. coli O104:H21 affecting 18 individuals and an outbreak of gastrointestinal illness, including bloody diarrhea, associated with E. coli O111:H8 in 56 persons (CDC 2000). Non-O157 serotypes of E. coli including O26:H11, O111:H8, O103:H2, O113:H21 and O104:H21 have been responsible for a small number of outbreaks in other parts of the world (CDC 1995b; Goldwater and Bettelheim 1995; Paton et al. 1996; Robins-Browne et al. 1998). In a cluster of three cases of HUS caused by O113:H21 in Australia, this organism was found not to have the attaching-and-effacing gene (Paton et al. 1999). 1.2.6 Sources and Modes of Transmission Fecal-oral transmission is the most common mode. For E. coli O157:H7, ingestion of contaminated food or direct contact with animals on farms or at petting zoos is common. Undercooked beef (especially hamburger), foods cross-contaminated from raw beef, and raw milk contaminated with cattle feces are the prototypical sources of common-source outbreaks. Venison is another potential source. Contaminated products, including leafy greens, alfalfa sprouts, and unpasteurized apple cider are other recognized exposure sources. Person-to-person transmission can occur directly (households, child care centers, institutions) or indirectly (contaminated drinking or recreational water). In all of these modes of transmission, the infectious dose is very low. Incubation Period uncommon) 18 days; usually 26 days (longer incubations are possible but

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1.2.7 Symptoms of E. coli infection: E. coli infection occurs when a person ingests Shiga toxin (Stx)-producing E. coli (e.g., E. coli O157:H7) after exposure to contaminated food, beverages, water, animals, or other persons. After ingestion, E. coli bacteria rapidly multiply in the large intestine and bind tightly to cells in the intestinal lining. This snug attachment facilitates absorption of the toxin into the small capillaries within the bowel wall, where it attaches to globotriaosylceramide (Gb3) receptors. Inflammation caused by the toxins is believed to be the cause of hemorrhagic colitis, the first symptom of E. coli infection, which is characterized by the sudden onset of abdominal pain and severe cramps, followed within 24 hours by diarrhea (Boyce, Swerdlow, & Griffin, 1995; Tarr, 1995). Hemorrhagic colitis typically occurs within 2 to 5 days of ingestion of E. coli, but the incubation period, or time between the ingestion of E. coli bacteria and the onset of illness, may be as broad as 1 to 10 days. As the infection progresses, diarrhea becomes watery and then may become grossly bloody, that is, bloody to the naked eye. E. coli symptoms also may include vomiting and fever, although fever is an uncommon symptom. On rare occasions, E. coli infection can cause bowel necrosis (tissue death) and perforation without progressing to hemolytic uremic syndrome (HUS)a complication of E. coli infection that is now recognized as the most common cause of acute kidney failure in infants and young children. In about 10 percent of E. coli cases, the Shiga toxin attachment to Gb3 receptors results in HUS. HUS had been recognized in the medical community since at least the mid-1950s; however, the syndrome first caught the publics attention in 1993 following a large E. coli outbreak in Washington State that was linked to the consumption of contaminated hamburgers served at a fast-food chain. A total of 501 E. coli cases were reported; 151 were hospitalized (31 percent), 45 persons (mostly children) developed HUS (9 percent), and three died (Bell, et al., 1994).During HUS, the majority of the toxin gains access to the systemic circulation where it becomes attached to weak receptors on white blood cells (WBC) thus allowing the toxin to ride piggyback to the kidneys where it is transferred to numerous strong Gb3 receptors that grasp and hold on to the toxin. Organ injury is primarily a function of Gb3 receptor location and density. These receptors are probably always in the gut wall and kidneys, but heterogeneously distributed in the other major body organs. This may be the reason that some patients develop injury in other vital organs (e.g., brain, etc). Once Stx attaches to receptors, it moves into the 12

cells cytoplasm where it shuts down the cells protein machinery resulting in cellular injury or death, and subsequent damage to vital organs such as the kidney, pancreas, and brain. 1.2.8 Diagnosis: Food quality/safety indicator tests Aerobic Plate Count The aerobic plate count (APC) or standard method count (SPC) is important in food microbiology as an indicator of the microbiological quality as well as a measure of sanitation used during handling of a food (Ray, 2004). APC determines counts of the non-fastidious aerobic bacteria. In some foods, high APC may indicate poor quality. Higher bacterial numbers spoil the food faster and result in loss of quality. Food which appears normal may have high APC, indicating that the food is about to spoil. In fresh products, APC indicates the effectiveness of sanitary procedures used during processing and handling and before storage of the product. A high APC in food products that were given heat treatment such as pasteurized milk may indicate that both shelf life stability and safety is affected (Ray, 2004). Although APC is a fast and efficient method to test the microbiological quality of the food, the test has some limitations (Yousef and Carlstrom, 2003). Fermented foods (e.g. cheddar cheese) naturally contain a high microbial load, and in consequence APC cannot be used to evaluate their general microbiological quality. In addition, the plating medium does not support the growth of fastidious microorganisms, which result in an under representation of these microorganisms in the APC. Moreover, incubation conditions favor growth of mesophilic aerobic bacteria, and other categories such as strict anaerobes are ignored. Standards for quality indicator tests for the various foods available are very limited. In fact, milk is the only food included in the U.S. federal standards which state that the APC is not to exceed 2 x 104 CFU/ml (CDFA, 2006). However, there is one study conducted in United Kingdom which suggests guidelines for ready-to-eat foods at the point of sale. There are different acceptance levels of APC for different food products. For example, sliced cooked ham has a suggested level of < 106 CFU/g as a satisfactory level, 106 - <107 as an acceptable level and 107 as unsatisfactory level (Gilbert, et al., 2000). 1.2.9 Public health significance: 13

1.2.9.1 Sources of E. COLI O157:H7 contamination Human infection is associated with the consumption of a number of contaminated foods among them meat, especially undercooked ground beef, raw milk, yoghurt, salamis, cheese and unpasteurized apple cider and water. Human beings and cattle carry the pathogen in their intestines and faeces hence a source of contamination to foods, water and the environment. The faeces and the bacteria may contaminate the udders and milking equipment and get into the milk during milking if adequate hygiene practices are not observed (Arimi et al., 2000). 1.2.9.2 Person-to-Person transmission of E. coli Outbreaks of E. coli O157:H7 can also be caused by person-to-person transmission, which has occurred in daycare centers, hospitals, nursing homes, and private residences. Because the infectious dose is so small it is very easy for the bacteria to be transmitted among people with close physical contact.

1.2.9.3 Prevention of E. COLI O157:H7 contamination through use of colicinogenic E. COLI strains: Several beneficial E. coli strains and lactic acid bacteria have the potential to produce colicins which reduce the shedding of E. coli O157:H7 or prevalence in cattle. Other interventions to reduce the prevalence of E. coli O157:H7 in cattle include the use of vaccination, feed additives, diet shifts, other antagonistic bacteria, and bacteriophages (Schamberger et al., 2004). 1.2.9.4 Prevention of E.COLI O157:H7 contamination through application of HACCP Quality Assurance practices important in prevention of microbial contamination of milk and milk products include implementation of HACCP in the manufacturing industry, and adherence to good husbandry practices. These include maintaining pastures free of harmful substances, 14

and water of potable quality. In order to avoid spread of infections within the herd, good hygiene status of the animals should be maintained and any incoming stock quarantined as part of the HACCP (Sandrou et al., 2000).

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1.3 Aims and objectives 1.3.1 Aims The aim is to study E. coli contamination of meat in the meat market. 1.3.2 Objectives To collect meat specimen from Galkaio meat market for bacteriological culture. Isolation and identification of E. coli colony. To make awareness of meat workers about the risks of meat contamination.

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Chapter two: Material and methods 2.1 Study site

Fgure1: map of Somalia indicating study area. The study area was concentrated around Galkaio meat market, between 17 May-15 June 2012. Galkaio, the capital of Mudug region, lies 750 km south of Bossaso. Galkaio district comprises of three food economy zones; 25% of the population falls within the Addun pastoral food economy zone and depends on mixed shoats and camel for their livelihood; 50% of the population falls within the Hawd pastoral food economy zone and depend on camel and shoats for a livelihood; 25% of the population falls within the urban food economy zone. The town has an estimated population of 70,000 inhabitants that is believed to be increasing with the influx of returnees and displaced people. The populated area covers about 5.9 km2 with the greatest density found adjacent to the main north-south road and airport road. Galkaio is divided into four main sections: Israac, Garsoor, Hormar in the northern part with an estimated population of 45,000; and Wadajir in the southern part. The demarcation line green line runs through the town and divides the town to two autonomous parts: North and South Galkaio.

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Located at an intersection crossroad area, Galkaio is a thriving trade town. Goods from the Bossaso port in the north, Berbera in the northwest, agricultural products from the south and livestock from Haud (zone 5) area of Ethiopia all find their way and exchange hands in Galkaio. Although there are two different local administrations in the north and south of the town, the collaboration of the two administrations enables local people, mainly women, cross the green line in pursuit of trade.

2.2 Material and Methods 50 out of 270 of butchers were selected from Galkaio Meat Market who is registered from the municipality of the district. 200 samples were collected, which is consisting of 100 meat samples and 100 samples from equipment (like: tables, knifes, axes and containers). The specimen was taken from the same 50 butchers for four different days. Specimen was taken by using swabs with transport media (BPW). Samples were properly labeled with the date of collection, source and type of material and species. Then, samples are placed in ice boxes and send to the Galkaio Central Veterinary Laboratory. Specimen is aseptically withdrawn from 1 ml with sterile pipettes. 1ml from each dilution, (10-1, 10-2 10-3 ), weigh a certain amount of Lauryl Sulfate Tryptose Broth (LSTB) base to distilled water mix well and distribute in to test tubes fitted with inverted Durhams tube and incubate at 30c or 37C for 24hrs. If neither gas formation nor opacity preventing the detection of gas formation is observed at this stage continue incubation for another 24 hrs, but, if there is gas formation or opacity is observed then go to the next step, because that is indicative of coliforms positive. A tube of brilliant green lactose bile broth is inoculated with 1ml from a tube of LSTB base showing opacity and or gas formation for confirmation and incubate at 30C or 37C for 24hours. The presence of coliforms is confirmed in case that gas formation has been noted after examination of the tube. (See the figure BGBB and LTB Media).

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BGBB (confirmation media)

LTB (selective enrichment media).

2.3 E. coli confirmation Inoculate 1ml from positive brilliant green lactose bile broth (BGBB) in to tryptone water. Incubate the tryptone water at 44c for 24 hr. test for indole production by adding 0.2-0.3ml of kovacs reagent to the tryptone water bottles, the development of a red ring color indicates the presence of E. coli.

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Chapter three: Result and Discussion 3.1 Result This study found Escherichia coli contamination of Galkaio Meat market. The 100 samples of meat 12 were found positive for E. coli. Whereas 100 samples of equipment 15 were found positive (see the Table). Types of Samples Meat Tables Knifes Axes Containers TOTAL No. of Samples 100 25 25 25 25 200 No. of Positive 12 6 4 4 1 27 Percentage (%) 12.00 24.00 16.00 16.00 4.00 13.50

The level of contamination of E. coli was found higher in equipment than in meat. This may indicate that the level of contamination is high and relatively risky. None of the butchers in the market had appropriate facilities for sanitary measures. Furthermore butchers and meat vendors in the market was unaware of risk associated with poor hygiene. During this study in Galkaio meat market 100 respondence in the study area was interviewed. Most of the respondence should lack of Good hygiene practices concept (see the figure below)

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100 80 60 40 20 0 D A F O IN E VIE AT R M T R W No ideaa bout GH P yesa bout GH P

Figure2: figure of Good hygiene practice concept According our result 90% of respondences indicates that they no idea about Good hygiene practice, and 10% of the respondences was aware because they attended some training about hygiene practices. Most workers was females except for camel meat sellers. Based on visual assessment and my experience the hygiene status of the containers and workers themselves was questionable, and no cooling system were used at all. The picture below shows the positive result is indicated by the red layer at the top of the bottles after the addition of kovacs reagent.

Figure 3: Indicates the positive result of an indole test in tryptone water

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3.2 Discussion The overall contamination of E. coli in the study area is found to be 15% in Equipment and 12% in Meat. similar findings were reported by Darwish et al.(2008) in the Amhara region of the east Gojjan zone in Ethiopia with the contamination of 17% in Equipment and 14% in Meat. Direct comparison of results is difficult due to differences in the study methodologies, such as the type of slaughtering, improved enrichment and isolation procedures, differences in sample size, the type of sample and how and when it was collected .

As shown in the finding of the present study, E. coli was detected from all items in the market, this indicates that the level of contamination is high. The contamination of E. coli pathogen may affect the health status of the customers of Galkaio. Equipment was found highly contaminated than the meat; this may indicate that there are no proper hygienic measures applied in the market and its premises at the end of the day routinely. The Galkaio meat market was poor and not properly arrangement as a meat market. The meat of the domestic species (camel, sheep and goats) are in the same table or shelf. This, however may increase the cross contamination among species (see the figure). In addition, meat worker are not trained and their health status are not totally controlled. This may also increase disease transmission through contamination and or direct disease transmission. The market is acclimatized full of house fly and stray cats.

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Figure4: indicates general view of the Galkaio meat market Slaughterhouse and meat market workers in Somalia are not well aware of food safety issues; Good Manufacturing Practice (GMP), hazard analysis critical control point (HACCP) system, and quality control is often not fully practiced. Factors could also contribute to the presence of E. coli: Manual slaughtering of animals using contaminated equipment. In adequate hygienic practices in slaughterhouse as well as meat market environment Transportation facilities Poor hygiene equipment in market places and slaughterhouse Poor personal hygiene in the study area

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Chapter four: Conclusion and recommendations: 4.1 Conclusion: From the obtained results, we can conclude that contamination by Escherichia coli was found in equipments and meat samples collected from Galkio meat market.

4.2 Recommendations: 1. Awareness of meat workers about personnel hygiene. 2. Careful handling and thorough cooking of raw and frozen meat, regardless of market source by the consumers is required to prevent food borne illness. 3. Municipality should improve slaughterhouses and meat market places. 4. Government should implement continuous monitoring of bacterial level in the slaughterhouse and markets so as to limit prevalence of E. coli. 5. Finally, Hazard Analysis Critical Control Point (HACCP) procedures should adopted during all steps of manufacture, handling as well as storage of meat products to produce safe and high quality products as well as ensuring compliance with legislation.

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List of References Adak GK, Meakins SM, Yip H, Lopman BA, O'Brien SJ. Disease risks from foods, England and Wales, 19962000. Emerging Infectious Diseases, 2005 March [cited2009August18].Availablefromhttp://www.cdc.gov/ncidod/EID/vol11no03/040191.htm. Clarence SY, Obinna CN, Shalom NC. Assessment of bacteriological quality of ready to eat food (Meat pie) in Benin City metropolis, Nigeria. Afr. J. Microb. Res., 2009; 3(6): 390395 CDC (2000). Escherichia coli O111:H8 outbreak among teenage campers - Texas, 1999. MMWR49(15):321-324. CDC (2000b). Food Net surveillance report for 1999 (final report) November. Gilbert, R.J., J. Louvois, T. Donovan, C. Little, K. Nye, C.D. Ribeiro, J. Richards, D. Roberts, and F.J. Bolton. 2000. Guidelines for the microbiological quality of some ready-to-eat foods sampled at the point of sale. Communicable Disease and Public Health, vol. 3, number 3, 163-167. ICMSF, 1996. International Commission on Microbiological Specifications for Foods Microorganisms in Foods 5. Characteristics of Microbial Pathogens. 1st Edn., Aspen Publishers. ISBN: 978-0-412-47350-0. Magnus P. Meat Composition. Food Science and Technology, 4th edition. Gohumunary Pub. London, 1981; pp. 108- 215. Paton JC, Paton AW (1998) Pathogenesis and diagnosis of shiga toxin-producing Escherichia coli infection. Clin Microbiol Rev 11:450-479. Ray, B. 2004. Fundamental Food Microbiology. 3rd. edition. CRC Press. 608p. Rao VA, Thulasi G, Ruban SW. Meat quality characteristics of non-descript buffalos as affected by age and sex. World Applied Science Journal, 2009; 1058-1065 Sandrou D, Arvanitoyannis. Implementation of HACCP to the dairy Industry: Current Status and Perspectives. Food Rev Int. 2000; 16:77-8. Schlundt J (2001) Emerging food-borne pathogens. Biomed. Enviro. Sci. 14(1-2):44-52. Tauxe R. V. (1997) Emerging Foodborne Diseases: An Evolving Public Health Challenge Emerging Infect. Dis. 3(4). WHO (1998) Zoonotic non-O157 Shiga toxin-producing Escherichia coli (STEC). Report of a WHO Scientific working group meeting. Berlin, Germany 23-26 June. Yousuf AHM, Ahmed MK, Yeasmin S, Ahsan N, Rahman MM, Islam MM. Prevalence of Microbial Load in Shrimp, Penaeus monodon and Prawn, Macrobrachium rosenbergii from Bangladesh. World Journal of Agricultural Sciences, 2008; 4 (S): 852-855 Yousef, A.E. and C. Carlstrom. 2003. Food Microbiology: A laboratory manual. John Wiley and Sons, Inc., Hoboken, New Jersey. 277p.

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Zaki-Eman, M.S., 1990. Escherichia coli in meat products with special reference to Enteropathogenic strains. M.V.Sc. Thesis, Faculty of Veterinary Medicine, Cairo University, Egypt.

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Annexes Questioner format:

1. 1 2 2. 1 2 3.

Do you get any training about meat hygiene practice? Yes No Do you know anything about hygiene practice during meat handling? Yes No If yes explain how

_____________________________________________________________________________ ___________________________________________________________________ 4. 5. 1 2 6. If Can you explain how you do it? Have you ever heard meat contamination? Yes No yes explain how

_____________________________________________________________________________ ___________________________________________________________________ 7. 1 2 8. If Do you know E. Coli meat contamination? Yes No yes explain

_____________________________________________________________________________ ___________________________________________________________________ 9. Do you know sanitation methods of meat dispensing and displaying?

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1 2 10. If

Yes No yes

explain_______________________________________________________________________ ___________________________________________________________________ 11. 1 2 Do you know E. Coli can transmit through meat handling? Yes No

Schedule of the study Date No.of samples No.of meat No.of table No.of hook No.of knife No.of axes samples 28

20/5/2012 23/5/2012 26/5/2012 29/5/2012 1/6/2012 3/6/2012 6/6/2012

15 25 30 25 35 30 40

samples 10 15 18 13 16 8 20

samples 5 3 7 2 4 10 6

samples 3 4 6 2 9 1 4

samples 2 5 1 6 3 2 3

3 1 2 1 1 3 1

E. coli thesis writing by Ali Mohamed Ali Iye

29

BGBB Media

sample register

Galkaio central laboratory

LTB Media

Positive Result

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