Protozoan Toxicity Tests

Protozoan toxicity test

The PROTOXKIT F contains all the materials necessary to perform six chronic (multigeneration) toxicity tests with the freshwater protozoan ciliate* Tetrahymena thermophila.

Ciliates are a very important component of benthic micro fauna communities and play a key role in the recycling of organic matter. They are also next to bacteria, the second most important group of biota in activated sludge of wastewater treatment plants.

Easy to follow instructions and detailed illustrations are provided in the kit for the conduct of range-finding and definitive acute tests, with calculation of the 24h EC50. The test organisms are included in the kits in a steady state condition in a very specific medium, which allows for their survival for several months at room temperature without prejudice to their good physiological condition.

Product No. TK11 PROTOXKIT F



Please review EBPI's step-by-step presentation on the Protoxkit F Procedure: 


protoxkit FullProtocol Instructions

Click here for Instructions for use.

Comparison Data of MicroBioTests

comparison Data MicroBiotests

Click here for comparison data.

Protozoan Toxicity Test Short Protocol


Click here to download the latest version of the PROTOXKIT F short protocol.

Test principle and test criterion

  • The assay is based on the measurement of the turnover of food substrate into ciliate biomass by optical density (OD) readings.
  • Inhibited culture growth under toxic stress is reflected by remaining turbidity of the food suspension (and hence higher OD) in comparison to the control.
  • Mean growth inhibition after 24h exposure at 30°C is determined and expressed as 24hEC50. NOEC and LOEC values can be calculated with specific data treatment program.


  • Uniform axenic stock cultures of high quality produced in strictly controlled conditions preclude variability associated with maintenance of live stocks in different laboratories.
  • The PROTOXKIT F uses standard disposable spectrophotometric cells as test containers, with direct measurement of the OD of the substrate in the cells.
  • A Quality Control Test with a reference chemical is described in detail, for accuracy and reproducibility check.


  • Tests can be started from the stock culture vial, eliminating the need and the costs of continuous culturing and maintenance of test organisms.
  • Minimal equipment needed for test performance: colorimeter or spectrophotometer,small incubator, conventional laboratory glassware.
  • Shelf-life of stock culture guaranteed for several months when stored properly, reducing test scheduling constraints.


  • Stock culture vial, sterile syringes for the transfer of the ciliates, tubes with substrate and reconstitution medium, stock dilution and test spectrophotometric cells and cell holders.
  • Detailed Standard Operational Procedure brochure, abbreviated Bench Protocol, data scoring sheets and graphical EC50 calculation sheets.
  • Specification sheet with batch number of the ciliate stock and the media.


  • Bioassays can be started anytime immediately from the stock culture.
  • Test performance only implies simple photometric OD readings at the start and at the end of the test.
  • Total performance time of the assay is approximately 1 hour.
  • A floppy disc for easy Toxkit data treatment can be obtained on demand.


  • Comparable to that of other ciliates in activated sludge and hence good indicator for toxic hazard in influents of wastewater treatment plants.
  • Compares favourably with that of human and animal cell lines pointing to the potential use of the PROTOXKIT F as a screening test in human toxicology.


Growth inhibition NOEC’s determined with PROTOXKIT F tests are ecologically meaningful thresholds for the protection of protozoan micro fauna in benthic communities and the well-functioning of the ciliate component in activated sludge.

Optional Components for the Chronic PROTOXKIT F

  • TK11  Chronic PROTOXKIT F with Tetrahymena thermophila freshwater tests (6 tests per kit)

Test Biota

  • TB11     5 vials with Tetrahymena thermophila (freshwater tests)

Test Containers

  • TC11     1 box with 100 spectrophotometric test cells + caps
  • TC12     10 stock culture calibration cells with caps
  • TC13     10 test tubes for ciliate inoculum

Test Media

  • TM11     10 Sets of concentrated media for fresh water toxicity tests ( 10 X 1 EPA medium)

Test Foods

  • TF13     10 tubes with food substrate & reconsitution medium for protozoan tests

Test Items

  • TI15     10 syringes for protozoan tests
  • TI17     10 cell holders for protozoan tests

Equipment Needed

  • Incubator with lateral illumination
  • Light table with traspartent stage
  • Luxmeter
  • Dissection microscope with transparent stage
  • Spectrophotometer