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Survival of Selected Bacteria Causing Traveler’s Diarrhea in Common Water and Beverages Presented By: Majed Al-Mashjary Ammar Basabrain Mohammad Raml Majed Al-Shaer Supervised By: Dr. Abdulaziz Bamarouf Outlines • Introduction about Traveler’s Diarrhea • Aims of this Project • Materials & Methods • Results • Discussion • Conclusions What is Traveler’s Diarrhea? • Traveler’s diarrhea is a syndrome characterized by a two time or greater increase in the frequency of unformed bowel movements • Commonly associated symptoms include abdominal cramps, nausea, urgency, fever, and malaise • Cases of Traveler diarrhea usually begin by surprise (sudden onset ) and occur during travel or soon after returning home and are mainly selflimited condition Traveler’s Diarrhea Can Cause More than a Stomachache! Common Signs and Symptoms Upset stomach Vomiting Fever Diarrhea Dehydration Don’t Count on these to Test for Food Safety! Sight Smell Taste Who is at risk? • Our immune system helps in fighting Infections. • But the immune systems of very young children, pregnant women, the elderly, and chronically ill people are at greatest risk to develop diarrheal infections Infants and young children, in particular, produce less acid in their stomachs, making it easier for them to get sick For pregnant women, the fetus is at risk because it does not have a fully developed immune system For elderly individuals, poor nutrition, lack of protein in the diet, and poor blood circulation may result in a weakened immune system Those who are immunocompromised, such as diabetics, cancer patients, AIDS patients, and people on antibiotics are at greater risk Etiological Agents of Traveler’s Diarrhea • Bacterial causes • Parasitic causes • Viral causes Bacterial Causes • The majority of cases of traveler’s diarrhea are caused by enteric bacterial pathogens, which are most likely acquired by ingesting contaminated food or water Salmonella • S. typhi which cause Typhoid fever. • Typhoid fever is an important cause of morbidity in many regions of the world, with an estimated 12 to 33 million cases occurring yearly Shigella • The etiological agent of Shigellosis is a highly contagious and severe inflammatory diarrhea disease • Sh. flexneri is the predominant species responsible for endemic disease • Each year there are about 160 million cases of shigellosis resulting in more than one million deaths Other Bacterial Etiological Agents Pathogenic Escherichia coli Campylobacter spp. Vibrio spp. Aeromonas spp Bacterial Food Poisoning • Bacterial food poisoning may in some circumstances be an important cause of diarrhea that typically occur in outbreaks • Staphylococcus aureus • Bacillus cereus • Clostridium perfringens Distribution of Traveler’s Diarrhea in the World • In general, countries with higher standard of hygiene have lower risk for traveler’s diarrhea WWW.CDC.GOV Water and Beverages Precautions • Water • Many pathogens causing traveler’s diarrhea have proved their ability to cause disease through tap water. • Tap water in many developing countries is often hazardous, particularly in rural areas or after rainfall • Ingestion of pathogens may occur by swallowing water while showering or by using tap water for brushing teeth • Ice cubes is one way of ingestion of untreated tap water specially with a pathogen of a low infectious dose such as Shigella spp • Finally, swimming in water that is contaminated by sewage should be avoided • Non-carbonated bottled water • Although non-carbonated bottled water is often used as an alternative, it is only as safe as the source from which it was taken • The viability of pathogens can be maintained in non-carbonated bottled water; in some cases, this vehicle has been a contributor to epidemics of cholera and typhoid fever • Tea and coffee • Tea and coffee are safe if consumed hot, whereas ice tea or coffee is not safe except if prepared commercially and bottled • Carbonated beverages • Carbonated beverages have the advantage that they are bactericidal • Carbonation result in an acid pH that will effectively kill bacteria over a period of time • Commercially bottled carbonated beverages are highly recommended as a fluid source for travelers • Unpasteurized milk • Unpasteurized milk and milk products are to be avoided entirely, including ice cream and small quantities of milk in tea or coffee • Finally , travelers have to be note that some beverage consumed in great quantity can contribute to diarrhea either through the action of caffeine or the effect of high sugar like fructose • Diet drinks containing aspartame (artificial sugar) may also produce diarrhea Aims of This Project 1) To determine the survival of common bacterial pathogens causing traveler’s diarrhea in: Water Carbonated drinks Fruit-based carbonated drinks Energy drinks Fresh juices 2) To measure the pH of all water and beverages used in this study Materials & Methods • Bacterial Source: Salmonella typhi Shigella flexneri clinical isolates from culture collection of microbiology laboratory in the Faculty of Medicine, King Abdulaziz University • Agar Media Nutrient Agar (Oxoid, UK) Drinks Ingredients 1) Carbonated Drinks: • Pepsi • Diet Pepsi Ingredients: • Carbonated water • phosphoric acid • Sugar (aspartame in diet) • natural cola flavored • caffeine 2)Carbonated Flavored Drinks: • 7up • Diet 7up Ingredient: • Carbonated water • citric acid • natural citrus flavored • Sugar (aspartame in diet) • Na citrate • Na benzoate 3) Energy Drinks: • Power horse • Diet Power horse • Bison • Red Bull Ingredients: • Carbonated water • citric acid • Sugar (aspartame in diet) • caffeine • Na citrate • Vitamins 4) Water: • Tap water • Bottle water • Evian water • Volvic Flavored water Ingredients: • Cations • Anions • * For volvic additional • Citric acid • Natural flavor 5) Fresh Juices: • Fresh lemon juice without sugar • Fresh lemon juice with sugar • Fresh orange juice without sugar • Fresh orange juice with sugar • pH measurement: • pH of all water and beverages were measured using standard pH meter • Procedure Determination of the survival of Salmonella and Shigella in different water and beverages drinks 1- For each bacterium, 14 separate sterile tubes were labeled with each water and beverages drinks 2- Five (5) ml of each water and beverages drinks were added to each tube 3- One McFarland standard of each bacterial suspension was prepared using sterile physiological saline 4- Then 100 µl of bacterial suspensions were added to each of the labeled tubes containing water and beverages drinks 5- All bacterial/drink mixtures were mixed well and left to stand for 30 minutes at room temperature 6- Then fixed amount from each mixtures were cultured in separate Nutrient agar medium by streaking on agar plates to grade bacterial survival 7- Then all agar plates were incubated for 24 hours at 37° C aerobically 8- The remaining bacterial/drink mixture (from step 5) were re-incubated for further 24 hours and then cultured again 9- All plates were examined for bacterial survival by interpretation of the results using streaking in agar plate’s method as follow: 0 = no growth +1= growth only in primary streak +2 = growth in the first and second streak. +3 = growth in the first, second and third streak +4 = growth in the first, second, third and fourth streak Pe ps i Pe ps i 7u p Die t7 po up we rh D.P ors ow e er ho rse Bis Bo on ttle wa ter Ta pw ate Ev r ian wa Fla ter vo ur wa Le ter mo nn oS Le mo nW Or /S an ge no Or S an ge W/ S Die t Survival of S. typhi in Water & Beverages 4 Growth in 30 min 3 Growth in 24 hr. 2 1 0 P Die ep s tP i ep si 7 Die up Po t7 up D. wer h Po we orse rh ors e Bis Re on d Bo ttle Bull w Ta ater p Ev wat e i Fa an w r lvo a ur ter Le wat er mo n Le mo noS nW Or an ge /S Or no an ge S W/ S Survival of Sh. flexneri in Water & Beverages 4 Growth in 30 min 3 2 1 0 Growth in 24 hr. Pe ps Die i tP ep si 7u p Die t7 Po up we rh D. ors Po e we rh ors e Bis on Re dB Bo ull ttle wa ter Ta pw ate Ev r ian wa Fa ter lvo ur wa ter Le mo nn oS Le mo nW Or /S an ge no Or S an ge W/ S pH Measurement of Water & Beverages 9 8.5 8 7.5 7 6.5 6 5.5 5 4.5 4 3.5 3 2.5 2 1.5 1 0.5 0 pH Discussion •The main aims of this project were to determine the survival of S. typhi and Sh. flexneri in different water and beverages and to measure pH of all drinks •As results indicate that some water and beverages were not bacteria friendly and bacteria die after either 30 min or 24 hours The bad side of soft drinks 1 - The high sugar content and low dietary value which is propriety of junk drink 2- Their affect on dental erosion 3- Their caffeine content and aspartame which affects the body The antibacterial killing of lemon juice as a natural drink is better than consuming soft drinks which have bad effect Conclusion Extensive consumption of one or other of these drinks may prevent illness Additional clinical and epidemiological studies are needed to further define the safety and suitability of drinks for consumption by foreign travelers. Acknowledgement • We thank allah for guiding us to the right path. • We thank our supervisor for supporting us & being beside us till the very last minute. Dr. Abdullaziz Bamarouf • Also we thank our family for their patience & supporting us & for being there for us.