STUDY OF BACTERIAL AGENTS CAUSING DIARRHEA IN CHILDREN BELOW 5 YEARS OLD AT HOSPITAL TUNKU AMPUAN JEMAAH,

SABAK BERNAM

PUNITHA A/P TANAGOBAL

DIPLOMA IN MEDICAL LABORATORY TECHNOLOGY

Faculty of Allied Health Sciences

SEGi College Subang Jaya

2016

STUDY OF BACTERIAL AGENTS CAUSING DIARRHEA IN CHILDREN BELOW 5 YEARS OLD AT HOSPITAL TUNKU AMPUAN JEMAAH,

SABAK BERNAM

Mini thesis submitted in partial fulfillment for the Diploma in

Medical Laboratory Technology

PUNITHA A/P TANAGOBAL

Faculty of Allied Health SciencesSEGI College Subang Jaya

2016

ACKNOWLEDGEMENTS

I would like to express deepest gratitude to my supervisor Miss.Nursyuhada Ismail for her full support expert guidance, understanding and encouragement throughout my study research. Without her incredible patience & timely wisdom & council. My thesis work would have been a frustrating & overwhelming pursuit. In addition, I express my appreciation to Miss.Valarmathy Mathivannan & Miss.Balvinder Kaur for their thoughtful questions and comments were valued greatly. I would like to thank my colleagues, who have supported me throughout entire process, both by keeping me harmonious and helping me putting pieces together.

TABLE OF CONTENTS

Content Page

TITLE PAGE i

CERTIFICATE ii

ACKNOWLEDGEMENTS iii

ABSTRACT iv

TABLE OF CONTENTS v

LIST OF FIGURES vi

LIST OF TABLES vii

LIST OF SYMBOLS AND ABBREVIATIONS viii

LIST OF APPENDICES ix

CHAPTER 1 INTRODUCTION

1.1 Background Study1.2 Objective1.2.1 General objective1.2.2 Specific objective1.3 Hypothesis1.3.1 Null hypothesis1.3.2 Alternative hypothesis1.4 Problem statement

CHAPTER 2 LITERATURE REVIEW

2.1 Diarrhoea in children2.2 Common bacteria agent2.3 Made of transmission2.4 Treatment

CHAPTER 3 METHODOLOGY

3.1 Sample collection3.2 Stool FEME3.3 Identification of bacteria

GANTT CHART

REFERENCES

CHAPTER 1

INTRODUCTION

1.1 Background Study

Common bacterial causes of diarrhea include campylobacter, salmonella, shigella and Escherichia coli. Diarrhea caused by bacteria and parasites can be common when traveling in developing countries and is often called traveler's diarrhea.Diarrhoel disease is the second leading cause of death in children under five years old. Diarrhoea can last several days, and can leave the body without the water and salts that are necessary for survival. Most people who die from diarrhoea actually die from severe dehydration and fluid loss. Diarrhoea is usually a symptoms of an infection in the intestinal tract, which can be caused by a variety of bacterial, viral and parastic organisms. Infection is spread through contaminated food or drinking-water, or from person -to-person as a result of poor hygiene. Diarrhoea is a major cause of malnutrition, and malnourished children are more likely to fail ill from diarrhoea. The list of possibilities is long. It could be caused by a viral or bacterial infection. It might also be the result of a parasite, a course of antibiotics, or something he ate. Viral infection. Any number of viruses like the rotovirus, adenovirus, calicivirus, astrovirus, and influenza can cause diarrhea, as well as vomiting, abdominal pain,fever, chills, and achiness. Bacterial infection.Bacteria such as salmonella, shigella, staphylococcus, campylobacter, or E. coli can also be responsible for diarrhea. If your toddler has a bacterial infection, he may

have severe diarrhea, accompanied by cramps, blood in the stool, and a fever. (He may or may not be vomiting.) Toddler’s diarrhea the persistent diarrhea that lasts a few weeks is much less common than diarrhea caused by a stomach virus, which virtually all kids experience at least a few times before they reach school age. Toddler’s diarrhea is usually the result of an out-of-whack diet. It’s normally caused by a diet low in fats and proteins and high in sugar and fluids. Drinking too much juice is associated with toddler’s diarrhea. The specific germs that cause diarrhea can vary among geographic regions depending on their level of sanitation, economic development, and hygiene contaminate crops or drinking water.

1.2 Objective

1.2.1 General objective To identify bacteria agent causing diarrhoea in children below 5 years

old.

1.2.2 Specific objective

To study the rate of diarrhoea infection in children below 5 years old at Hospital Tunku Ampuan Jemaah, Sabak Bernam.

1.3 Hypothesis

1.3.1 Null hypothesis Salmonella sp. causing higher infection rate for diarrhoea in children

below 5 years old.

1.3.2 Alternative hypothesis Rotavirus causing higher infection rate for diarrhoea in children below

5 years old.

1.4 Problem Statement

In 2011, diarrhoea accounted for 700,000 deaths in children under five years of age in worldwide making it the second leading cause of child mortality. The highest rates of child mortality are in Sub-Saharan Africa and Southeast Asia. An estimated 1.7 billion episodes of diarrhoea, equaling approximately 2.9 episodes per child per year, created health

system costs of about 7 billion US dollars. The majority of diarrhoeal diseases can be prevented by implementing water, sanitation and hygiene programs, which all aim at interrupting faecal–oral transmission pathways. Even though diarrhoea morbidity and mortality has decreased since the 1990s, the overall disease burden remains unacceptably high, particularly in low and middle-income countries. Diarrhoea is a leading cause of malnutrition in children under five years old.

DR. Anil Gupta (2014)

CHAPTER 2

LITERATURE REVIEW

2.1 Diarrhoea in children

Diarrhoea disease is the second leading cause of death in children under five years old, and is responsible for killing around 760 000 children every year. Diarrhoea can last several days, and can leave the body without the water and salts that are necessary for survival. Diarrhoea is usually a symptom of an infection in the intestinal tract, which can be caused by a variety of bacterial, viral and parasitic organisms. Children who die from diarrhoea often suffer from underlying malnutrition, which makes them more vulnerable to diarrhoea. Each diarrhoeal episode, in turn, makes their malnutrition even worse. Diarrhoea is a leading cause of malnutrition in children under five years old. The majority of diarrhoeal diseases can be prevented by implementing water, sanitation and hygiene (WASH) programs, which all aim at interrupting faecal–oral transmission pathways, commonly referred to as the five “F” (fluids, fields, flies, fingers and food). Several studies have attempted to evaluate the effects of combined or single water, hygiene and sanitation interventions on diarrhoea as an outcome variable.

2.2 Common bacteria in children The most common cause is an infection of the intestine due to either a virus, bacteria or parasite; a condition known as gastroenteritis. These infections are often acquired from food or water that has been contaminated by stool, or directly from another person who is infected. It may be divided into three types which is short duration watery diarrhea, short duration bloody diarrhea and more than two weeks, persistent diarrhea. The short duration watery diarrhea may be due to an infection by cholera. If blood present it is also result in diarrhea, including hyperthyroidism, lactose intolerance, inflammatory bowel disease, a number of medications, and irritable bowel syndrome. In most cases, stool cultures are not required to confirm the exact cause. Causing the bacteria agent virus. Viruses that can cause diarrhea include Norwalk virus, cytomegalovirus and viral hepatitis. Rota virus is a common cause of acute childhood diarrhea. Common bacterial causes of diarrhea include campylobacter, salmonella shigella and Escherichia coli. Diarrhea caused by bacteria and parasites can be common when traveling in developing countries and is often called traveler’s diarrhea.

2.3 Mode of transmission

All of the more common childhood diarrhea caused by pathogenic bacteria & viruses are transmitted via the faecal oral route. The phategenes discharge in a faeces of an infected person may enter the body of another susceptible person through the mouth. This may occur among children ingesting food/ water contaminated in human excreta. Direct transmission among persons in close contact is also possible. Such transmission could occur via unclean hands or through contaminated objects such as bad linen kitchen utensils & tableware. Files & cockroaches play a role as vectors of the infections agent of faecal origin. Most of the pathogenic organisms that cause Diarrhea and all the pathogens that are known to be major causes of Diarrhea are transmitted primarily or exclusively by the faeco–oral route. Faeco oral transmission may be waterborne, food borne, or direct transmission which implies an array of other faeco oral routes such as via fingers, or fomites, or dirt which may be ingested by young children. Acute diarrhoeal diseases usually spread by contaminated hands or ingestion of contaminated food or drinks, and occasionally by aerosol. Outbreaks may occur in settings like institutions and child care centers.

2.4 Treatment

2.4.1 Probiotics

Probiotics contain healthy bacteria (lactobacilli) that can replace unhealthy bacteria in the gastro intestinal track.Yougurt is the easiest source of probiotics. Give children to two to six once of yogurt twice daily. Today, almost all yogurts are active culture which means that they contains life and active bacteria. Probiotic supplements in granules tablets or capsule are also can be given which available in health food stalls.

Medicine such as bismuth subsalicylate (Pepto-ismol, Kaopectate) or lope amide (medium) which are available as liquids/tablets. Follow the instructions on the package. If you have itching, burning, soreness or pain in your rectal area because you have a lot of bowel movements, try these tips to feel better. Take a warm both. Afterwards, pat the area dry (do not rub) with a clean, soft towel. Use a hemorrhoid cream or try white petroleum jelly. Do your best to stay hydrated.You should drink at least six 8-ounce glasses of fluid each day? Choose fruit juice without pulp, broth or soda (without caffeine).Chicken broth (without the fat), tea with honey or sports drinks are also good choice.

2.4.2 Prebiotics

Taking prebiotics for diarrhea relief is still an underutilized tool, but it is quickly gaining popularity as we learn more about how the digestive system functions. The digestive system contains a certain amount of beneficial bacteria and if the level of these bacteria gets too low, it can leave you susceptible to a number of uncomfortable conditions, including diarrhea. Maintaining the proper bacteria levels in your digestive system is precisely where prebiotics can be a big help. Prebiotics are a form of carbohydrate that does not break down as it passes through the digestive tract. It is then used as a food source by probiotics, or beneficial bacteria. It is this beneficial bacteria that can help you to fight off diarrhea. Prebiotics have been shown to have multiple benefits including increased absorption of calcium and magnesium increased immune system effectiveness inhibition the growth the lesions in the gut, such as adenomas and carcinomas, therefore reducing risk factors associated with diseases in the rectum and colon

CHAPTER 3

METHADOLOGY

3.1 SAMPLE COLLECTION

100 stool samples will be collected below 5 years old at Hospital Sabak Bernam.

3.2 Stool FEME

Fecal samples (one from each subject) from children without diarrhea were collected in a clean container. From the children with diarrhea, one stool specimen was collected within 24 hours of admission. All feces were collected in special containers with Cary–Blair transport medium, kept at 4 °C and transported to the microbiological laboratory within 24 hours. (Look under microscope.)

3.2 Stool culture

Stools will be cultured on MacConkey agar for the selection of Escherichia coli, TCBS agar for the selection of Vibrio species; and XLD agar for the selection of Shigella spp and Salmonella spp. A stool (feces) sample can provide doctors with valuable information about what's going on when a child has a problem in the stomach, intestines, or other part of the gastrointestinal (GI) system and incubate at body tempreture for 24 hours.A stool culture helps the doctor determine if there's a bacterial infection in the intestines. A technician places small stool samples in

sterile plastic dishes containing nutrients that encourage the growth of certain bacteria. The targeted bacteria will only grow if they're already present in the stool sample. If bacterial colonies form, the technician evaluates them using a microscope and chemical tests to identify the organism.

3.3 Identification of bacteria

3.3.1 Escherichia coli on Mac Conkey agar

Streak plate isolation of E. coli on MacConkey Agar grown for 24 hrs at 37 degrees. E. coli demonstrates strong lactose fermentation indicated by the bright pink halo, bile precipitant around the colonies, and pink colony growthThiosulfate-citrate-bile salts-sucrose agar or TCBS agar is a type of selective agar culture plate that is used in microbiology laboratories to isolate Vibrio spp. TCBS Agar is highly selective for the isolation of V. cholerae and V. parahaemolyticus as well as other vibrios. Xylose Lysine Deoxycholate (XLD) Agar was developed by Taylor for the differentiation, isolation, and identification of enteric pathogens, and to support the growth of more fastidious enteric organisms. XLD Agar was especially designed to allow the growth of Shigella species, and is a proven medium for the isolation of this organism. It has also been found to be an excellent medium for isolating Salmonella species as well. The disease is characterised by vomiting and watery diarrhoea lasting for three to eight days. Fever and abdominal pain occur frequently. Treatment is symptomatic. Maintenance of hydration is the most important measure. Diagnosis may be made by rapid antigen detection of rotavirus in stool specimens. Strains may be further characterised by enzyme immunoassay or reverse transcriptase polymerase chain reaction. Stools for these tests should be collected in the acute phase of illness.

GANTT CHART

November January February March April

Title Selection

Proposal preparation/Update literature review

Proposal Presentation

Data collection

Thesis Completion

Preparation for Final presentation

Final Presentation

 

REFERENCE

DR. Anil Gupta (2014). Study of the prevalence of diarrhoea in children under the age of five years: its association with wasting. Indian J.Sci.Res. 7 (1): 1315-1318, 2014

Katharina Diouf (2014). Diarrhoea prevalence in children under five years of age in rural Burundi: an assessment of social and behavioral factors at the household level. Retrieved from http://www.globalhealthaction.net/index.php/gha/article/view/24895.