Agricultural & EnvironmentalAgricultural & Environmental

Lab.

Water quality testing II:

PCR-based testing for water bacterial contaminants

The Islamic University

Faculty of Science

Biotechnology Dep.

غزة – االسالمية الجامعة

العلوم كلية

الحيوية التكنولوجيا قسم

EXPERIMENT OBJECTIVE:

The objective of this experiment is to use PCR to determine the quality of water and to detect and monitor the presence of bacterial contaminants in environmental bodies of water.

Water pollution is a major worldwide health problem and thus an essential subject area for incorporation into health education (specifically biology and ecology courses). According to the World Health Organization (WHO), over nine million deaths annually are attributed to waterborne diseases, occurring mostly in developing countries.

It is projected that 38% of those deaths occur in children afflicted with diarrhea. Furthermore, the WHO estimates that 80% of all worldwide infectious disease is due to unsafe water sanitation. Typical infections are cholera, giardiasis, hepatitis, shigellosis, typhoid, and acute gastrointestinal illness (AGI).

CholeraCholera

Cholera is an acute intestinal infection caused Cholera is an acute intestinal infection caused by ingestion of food or water faecally by ingestion of food or water faecally contaminated with the bacterium contaminated with the bacterium Vibrio Vibrio choleraecholerae.. It has a short incubation period, from It has a short incubation period, from less than one day to five days, and produces an less than one day to five days, and produces an enterotoxin that causes a copious, painless, enterotoxin that causes a copious, painless, watery diarrhoea that can quickly lead to severe watery diarrhoea that can quickly lead to severe dehydration and death if treatment is not dehydration and death if treatment is not promptly givenpromptly given. . Vomiting also occurs in most Vomiting also occurs in most patientspatients. .

giardiasis

The Life Cycle of Giardia Lamblia

Giardia lamblia

cyst

Entamoeba histolytica Life cycle

the cysts can survive days to weeks in the external environment and are responsible for transmission

Trophozoites passed in the stool are rapidly destroyed once outside the body

Entamoeba histolytica

CYST 10 – 15 µMtrophozoite15 to 20  µm.

cyst will have 1 to 4 nuclei. In addition, the peripheral chromatin is generally evenly distributed.

ShigellaShigella bacteria can contaminate food and bacteria can contaminate food and water supplies, especially in areas where the water supplies, especially in areas where the sanitation is not adequatesanitation is not adequate. . However, most of the However, most of the time the bacteria are spread when a person time the bacteria are spread when a person comes into contact with another person's stool comes into contact with another person's stool ((pooppoop). ). That's one of the reasons why it's That's one of the reasons why it's important toimportant to wash your handswash your hands after using the after using the restroomrestroom. . It takes as few as 10 of these bacteria It takes as few as 10 of these bacteria to cause an infectionto cause an infection. . That means thatThat means that ShigellaShigella bacteria can spread easily within families, bacteria can spread easily within families, schools, childschools, child--care centers, nursing homes, and care centers, nursing homes, and other institutionsother institutions

shigellosis

shigellosis

The bacteria produce toxins that can The bacteria produce toxins that can attack the lining of the large intestine, attack the lining of the large intestine, causing swelling, ulcers on the intestinal causing swelling, ulcers on the intestinal wall, and bloody diarrheawall, and bloody diarrhea. . Symptoms can Symptoms can range from just watery diarrhea to bloody range from just watery diarrhea to bloody diarrhea, fever, and abdominal paindiarrhea, fever, and abdominal pain

TyphoidTyphoid

Typhoid feverTyphoid fever is a bacterial disease, caused by is a bacterial disease, caused by Salmonella typhiSalmonella typhi. . It is transmitted through the It is transmitted through the ingestion of food or drink contaminated by the ingestion of food or drink contaminated by the faeces or urine of infected peoplefaeces or urine of infected people. .

Symptoms usually develop 1–3 weeks after Symptoms usually develop 1–3 weeks after exposure, and may be mild or severeexposure, and may be mild or severe. . They They include high fever, malaise, headache, include high fever, malaise, headache, constipation or diarrhoea, roseconstipation or diarrhoea, rose--coloured spots coloured spots on the chest, and enlarged spleen and liveron the chest, and enlarged spleen and liver. . Healthy carrier state may follow acute illnessHealthy carrier state may follow acute illness. .

Acute gastrointestinal illness AGIAcute gastrointestinal illness AGI

Acute gastrointestinal illness (AGI) is caused by a variety of different agents and conditions and comprises a constellation of symptoms that may include diarrhea, nausea, vomiting, abdominal pain, abdominal cramps, fever, and

other systemic symptoms. Because of its various causes and variable symptomatology, no standard definition of AGI has been presented in the medical literature, making comparisons

of studies difficult.

WHAT IS CRYPTOSPORIDIUM? Cryptosporidium parvum is a parasite that can

infect man and a wide range of domestic and wild animals. The organism can be acquired from contaminated food, milk, drinking water, poorly maintained swimming pools, farm animals and other infected people. It exists within the gut cells of a host and later in its life cycle is excreted in stools in the form of ‘oocysts’, the infectious form of the parasite.

oocysts of Cryptosporidium

parvum

PCRPCR