The document discusses the detection of Cryptosporidium, a spore-producing parasite found in contaminated food and water, and its transmission routes. It outlines the history, symptoms, and risk groups associated with Cryptosporidiosis, as well as methods for sample collection and parasite detection. Future research aims to identify different species of Cryptosporidium across various products.

2. Detection of Cryptosporidium in
Freshwater, Fruits, Vegetables and
Dairy products.
Muhammad Iqbal BS Microbiology
Govt College University Fsd.
Internship at CVAS

4. Contents
• History
• Introduction
• Aims and objectives
• Materials and methods
• Future perspectives

5. Introduction
• Cryptosporidium is a spore producing parasite
found in the intestine of infected people and
animals.
• Cryptosporidium spp is the most common
parasite in verterian students and farm
workers.
• By ingesting food or drinks contaminated with
fecal sample.

6. • Swallowing recreational water contaminated
with cryptosporidium.
• Not washing hands.
• Person to person transmission.
• Food borne transmission.
• Water borne transmission.
• Zoonotic transmission.
• Anthroponotic transmission.

8. Signs and Symptoms
• Symptoms (2 to 10 days after infection and
can last up to 30 days.)
• Diarrhea
• Stomach cramps
• Dehydration
• Nausea
• Vomiting
• Weight loss
• Sometimes no symptoms are seen.

10. Who is most at risk
• Children who attend day care centers, including diaper-
aged children
• Child care workers
• Parents of infected children
• People who take care of other people with
cryptosporidiosis
• International travelers
• Backpackers, hikers, and campers who drink unfiltered,
untreated water
• People who drink from untreated shallow, unprotected
wells.
• People, including swimmers, who swallow water from
contaminated sources
• People who handle infected cattle

11. History
• Recognized in mice in 1907.
• Reported in Human in 1976.
 Immunnocompetant child
 Immunosuppressed child
• Recognize globally in 1980s and 1990s.
 AIDS Patients
 Out break in verterian students.

12. Aims and objectives
• To determine the cryptosporidium infection in
 Fresh water
 Fruits
 Vegetables
 Dairy products.

13. Materials and Methods
• Collection of samples.
For fresh water sample from different taps
For fruits from fruit shops and rotten fruits
For vegetables from vegetable shops and field
For dairy from dairy shops and farms
• Make dilution of the samples.

14. • Direct smear preparation from fresh sample.
• formalin-ether method:
It increase chances of detection of parasitic
organism
Formalin fix the eggs, larvae,
Oocysts, and preserve the
morphology.

15. zehil-nelson method
• Prepare smear 1-2 drops of specimen and dry.
• Fix with methanol.
• Stain with carbol fuschin for 1 min.
• Destain with acid alcohol for 2 min.
• Counter stain with malachite green for 2 min.
• Dry slides.
• Examine 200-300 field using 40x objective.

18. Loop-Mediated Isothermal Amplification
• Four different primer used.
• Design to recognize six distinct regions.
• Reaction processed at constant temperature.
• Amplification and detection completed in
single steps.
• By incubating the of mixture samples, primers,
DNA polymerase with strand displacement
activity and substrates at constant
temperature.

27. Future perspectives
• Identification of types of cryptosporidium
species in different products.