Isolation of Nematodes and Identification of C.elegans, from soil
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CUBE Mumbai, TIFR CUBE Winter Meet 2016 Isolation procedure of nematodes and identification nematodes as C.elegans using microscopy.
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Isolation of Nematodes and Identification of C.elegans, from soil
- 1. Isolation of Nematodes and Identification of C.elegans Prepared and Presented by: - Shivam kumar Sriwas - Ashwini Singh - Manasi Pal - Vaidehi Joishy (Smt.C.H.M College,Ulhasnagar and CUBE Mumbai, TIFR) B.Sc. Biotechnology
- 2. Soil organisms…. Two major types :- A)flora and B)Fauna Soil Fauna:- It includes Nematodes , Protozoa , Earthworms and other organisms.
- 3. What is C.elegans? Caenorhabditis elegans is a free-living, non-parasitic, small soil worm or nematode that can be found throughout the world. It is transparent and ~1mm in size.
- 4. What are soil nematodes ? • The nematodes are also known as roundworms or threadworms which belong to phylum Nematoda. • They have cylindrical body with tapering ends. • There are no segmentations as such seen in earthworms. • They possess a strong but flexible outer layer cuticle.
- 5. What makes C.elegans a Model organism ? • Cell count of an adult C.elegans is exactly known i.e. it has 959 somatic cells and function of each group of cell is also known, and variable number of germ cells. • It has short life cycle of 3 days with large brood size. • It has simple and transparent anatomy.
- 7. Isolation of nematodes We prepared plain agar plates (2gms of agar in 100 ml of water). Then placed 2 drops of E.coli saline suspension and spread it at the center. Then placed the collected soil sample around the peripheral region(Half of the petri plate). Within one day we can observe the nematodes crawling out of the soil sample towards E.coli region.
- 8. STEP 2 :- SINGLE LINE CULTURE
- 9. Creating single line culture plate plain agar plates with single nematode & & E.coli plain agar plates with single nematode for seperation of eggs. {Pick up a single nematode & place it on another plain agar plate swabbed with E.coli.} plain agar plate with soil sample & E.coli
- 11. Classification of C. elegans Classification:- Phylum : Nematoda {Greek - nemat = thread ; –odes = resembling} Class : Chromadorea {Oesophagus divided into bulbs} Order : Rhabditida {Greek – rhabdos= a stick, for shape of buccal cavity} Family : Rhabditidae {bacteria feeders} Genus : Caenorhabditis {Greek - caeno- = new and rhabditis = rod-like } Species : elegans
- 12. ~Phylum : Nematoda • Thread like , cylindrical , Coelomate body. • Exhibits parasitism. • Tubular digestive system with opening at both the ends.
- 13. ~Class : Chromedorea • Oesophagus is divided into bulb with 3-5 oesophagal glands. • Closely related class : Enoplea – Oesophagus is cylindrical.
- 14. ~Order : Rhabditida • Stick like buccal cavity. • Delicate rays in caudal papillae.
- 15. ~Family : Rhabditidae • Males have 5 bilateral pairs in papillae.
- 18. Comparing what we get with C. elegans for identification • Presence of two bulb at pharynx region of C.elegans is first important & main feature to differentiate…..
- 19. Thank you…!!!