Protistiary - Protocols: Traps

PROTOCOLS Traps
Incomplete - still require images Picture of polystyrene blocks with slides in them Picture of tube with coverslips sticking out the side Picture of Sponge blocks Picture of protective bag Weighted string Collecting with a bucket An array of different materials can be placed in sediments or in the water column to act as traps for protists, and especially protozoa. The ‘traps’ are placed in a natural location for a period of time (2-5 days usually), and then removed and returned to the laboratory. These traps select subsets of organisms from the habitat, but typically develop complex and rich communities. It is wise to place the ‘traps’ inside a protective mesh bag or perforated box to prevent larger animals gaining access and browsing on the microbial populations. Nylon tights serve this role very well. You will also need access to a quiet undisturbed sites. Materials Cover slips Slides 4 inch thick-walled rubber tubing Sharp blade Fishing weights or other weights Covered jar or Ziploc bag Bucket Polystyrene packing material Sponge (plastic bath sponge) Nylon string / fishing line Nylons (pantyhose) Buoy (plastic drinks bottle) Plastic ziploc sandwich boxes Sealable plastic bags Sealable plastic containers - such as sandwich boxes forceps tape - for containers indelible pens - to write on tape notebook Some folk like to take notes, GPS readings, temperature, pH etc. etc. The longer traps are allowed to sit, the more different organisms will be observed. Allow traps to sit 24 -120 hours before removing. It is most important that the slide and coverslip traps do not break through water because this could strip the biological community off the surfaces. To do this carefully place a bucket alongside the rope but under water, remove the bags under water and place them in the bucket. Remove bucket. The samples can then be moved using the same technique into a bag or sandwich boxes, and sealed for return to the lab. In the laboratory, slides and coverslips are removed by holding them horizontally as they are brought out of the water. Clean and polish one face of the slides / coverslips before observing. To observe organisms from the styrofoam peanut trap, simply squeeze the styrofoam or polyurethane foam pieces into a dish. Observe dishes with a dissecting scope or pipette a drop of water onto a microscope slide and cover with a coverslip to observe under the light microscope.
Trap 1. Slides in polystyrene Cut two blocks approximately 1” X 1” X 3” from polystyrene packing material, with a razor blade make half inch deep cuts along the long faces about 1 cm apart. Insert slides with one block on either end of the slide Place in a pantyhose bag Attach to string, a line of these can be placed along the string / rope. One end of the string rope must have a float, such as a plastic drinks bottle, and the other an anchor. When collecting these traps, submerge a bucket alongside the traps, and move the traps into the bucket without pulling them through the surface of the water Remove each slide from the polystyrene blocks while the traps are submerged, life each slide horizontally from the water so that the surface tension does not strip the biofilm from the surface of the slide. Clean off the lower surface, and add coverslips to the upper surface, remove excess fluid before observing.	Image D. J. Patterson
Trap 2 coverslips Take 6 “ length of rubber tubing and make cuts alternately back and front Insert coverlsips into the tubing Place in a pantyhose bag Attach to string, a line of these can be placed along a string / rope. One end of the string rope must have a float, such as a plastic soda bottle and the other would usually be anchored. when collecting these traps, submerge a bucket alongside the traps, and move the traps into the bucket without pulling them through the surface of the water Remove each slide from the polystyrene blocks while the traps are submerged, life each slide horizontally from the water so that the surface tension does not strip the biofilm from the surface of the slide. Clean off the lower surface, and add coverslips to the upper surface, remove excess fluid before observing.	Image D. J. Patterson
Trap 3. Sponge blocks Cut a plastic bathroom sponge into blocks about 1” X 1” Place 4-6 in a pantyhose bag Attach to string, a line of these can be placed along the string / rope. One end of the string rope must have a float, such as a plastic soda bottle After collection, squeeze the sponge to release the organisms contained inside it. This technique has been widely used to assess the communities in habitats. However, the conditionsn inside the sponge blocks is very different to that of the surrounding medium, so the samples are neither representative of the surrounding water nor reflect the characteristics of the community in the surrounding water.
Trap 4. Styrofoam peanuts The tiny spaces in styrofoam peanuts and blocks and polyurethane foam can also be colonized by microorganisms and can be used to fish bacteria and protists out of marine and freshwater environments. Gather several pieces of string or monofilament fishing line (approximately eight inches long) together in a bundle and tie in a single knot at one end. To the end of each piece of string except one, tie a styrofoam peanut or a small piece of soft foam. Add a fishing weight to last piece of string and add a long string or fishing line leader to the top of the tied bundle. This array of traps can be tied to a pole and suspended in the water, or tied to a stone or brick and allowed to sink to the bottom of the water being sampled. Be sure that the foam is not exposed at low tide. The length of string leading to each foam piece should be short so that the foam does not float above the water surface.
Protistiary home page Top of Protocols

Incomplete - still require images

Picture of polystyrene blocks with slides in them
Picture of tube with coverslips sticking out the side
Picture of Sponge blocks
Picture of protective bag
Weighted string
Collecting with a bucket

An array of different materials can be placed in sediments or in the water column to act as traps for protists, and especially protozoa. The ‘traps’ are placed in a natural location for a period of time (2-5 days usually), and then removed and returned to the laboratory. These traps select subsets of organisms from the habitat, but typically develop complex and rich communities.

It is wise to place the ‘traps’ inside a protective mesh bag or perforated box to prevent larger animals gaining access and browsing on the microbial populations. Nylon tights serve this role very well.

You will also need access to a quiet undisturbed sites.

Materials

Cover slips
Slides
4 inch thick-walled rubber tubing
Sharp blade
Fishing weights or other weights
Covered jar or Ziploc bag
Bucket
Polystyrene packing material
Sponge (plastic bath sponge)
Nylon string / fishing line
Nylons (pantyhose)
Buoy (plastic drinks bottle)
Plastic ziploc sandwich boxes
Sealable plastic bags
Sealable plastic containers - such as sandwich boxes
forceps
tape - for containers
indelible pens - to write on tape
notebook

Some folk like to take notes, GPS readings, temperature, pH etc. etc.

The longer traps are allowed to sit, the more different organisms will be observed. Allow traps to sit 24 -120 hours before removing. It is most important that the slide and coverslip traps do not break through water because this could strip the biological community off the surfaces. To do this carefully place a bucket alongside the rope but under water, remove the bags under water and place them in the bucket. Remove bucket. The samples can then be moved using the same technique into a bag or sandwich boxes, and sealed for return to the lab. In the laboratory, slides and coverslips are removed by holding them horizontally as they are brought out of the water. Clean and polish one face of the slides / coverslips before observing. To observe organisms from the styrofoam peanut trap, simply squeeze the styrofoam or polyurethane foam pieces into a dish. Observe dishes with a dissecting scope or pipette a drop of water onto a microscope slide and cover with a coverslip to observe under the light microscope.

Trap 1. Slides in polystyrene

Cut two blocks approximately 1” X 1” X 3” from polystyrene packing material, with a razor blade make half inch deep cuts along the long faces about 1 cm apart.
Insert slides with one block on either end of the slide
Place in a pantyhose bag
Attach to string, a line of these can be placed along the string / rope. One end of the string rope must have a float, such as a plastic drinks bottle, and the other an anchor.
When collecting these traps, submerge a bucket alongside the traps, and move the traps into the bucket without pulling them through the surface of the water

Remove each slide from the polystyrene blocks while the traps are submerged, life each slide horizontally from the water so that the surface tension does not strip the biofilm from the surface of the slide. Clean off the lower surface, and add coverslips to the upper surface, remove excess fluid before observing.

Image D. J. Patterson

Trap 2 coverslips

Take 6 “ length of rubber tubing and make cuts alternately back and front
Insert coverlsips into the tubing
Place in a pantyhose bag
Attach to string, a line of these can be placed along a string / rope. One end of the string rope must have a float, such as a plastic soda bottle and the other would usually be anchored.
when collecting these traps, submerge a bucket alongside the traps, and move the traps into the bucket without pulling them through the surface of the water

Remove each slide from the polystyrene blocks while the traps are submerged, life each slide horizontally from the water so that the surface tension does not strip the biofilm from the surface of the slide. Clean off the lower surface, and add coverslips to the upper surface, remove excess fluid before observing.

Image D. J. Patterson

Trap 3. Sponge blocks

Cut a plastic bathroom sponge into blocks about 1” X 1”
Place 4-6 in a pantyhose bag
Attach to string, a line of these can be placed along the string / rope. One end of the string rope must have a float, such as a plastic soda bottle
After collection, squeeze the sponge to release the organisms contained inside it.

This technique has been widely used to assess the communities in habitats. However, the conditionsn inside the sponge blocks is very different to that of the surrounding medium, so the samples are neither representative of the surrounding water nor reflect the characteristics of the community in the surrounding water.

Trap 4. Styrofoam peanuts

The tiny spaces in styrofoam peanuts and blocks and polyurethane foam can also be colonized by microorganisms and can be used to fish bacteria and protists out of marine and freshwater environments.

Gather several pieces of string or monofilament fishing line (approximately eight inches long) together in a bundle and tie in a single knot at one end.
To the end of each piece of string except one, tie a styrofoam peanut or a small piece of soft foam.
Add a fishing weight to last piece of string and add a long string or fishing line leader to the top of the tied bundle.
This array of traps can be tied to a pole and suspended in the water, or tied to a stone or brick and allowed to sink to the bottom of the water being sampled. Be sure that the foam is not exposed at low tide. The length of string leading to each foam piece should be short so that the foam does not float above the water surface.