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Toxikologisk tillväxtstudie av sötvattenalgen Raphidocelis subcapitata: En jämförelse mellan flödescytometer NovoCyte och automatisk cellräknare TC20
Kristianstad University, Faculty of Natural Science.
2018 (Swedish)Independent thesis Basic level (degree of Bachelor), 10 credits / 15 HE creditsStudent thesisAlternative title
Study of toxic growth inhibition with unicellular fresh water green algae Raphidocelis subcapitata : Comparison between flow cytometer NovoCyte and automatic cell counter TC20 (English)
Abstract [sv]

Biologiska toxicitetstester utförs genom att exponera en testorganism för olika koncentrationer av kemikalier under en bestämd period. Det finns akuta toxicitetstester och kroniska. Resultat från akuta studier presenteras som EC50-värde (Effect Concentration, affecting 50 % of the population). Tester som används som underlag för riskbedömningar ska utföras på kvalitetsmässigt acceptabelt sätt som grundas på International Organization for Standardization (ISO) och Good Labaratory Practice (GLP). Syftet med studien var att studera toxisk inverkan av kaliumdikromat på encellig sötvattenalg, R. subcapitata, genom att räkna cellantal med två olika instrument: flödescytometer och automatisk cellräknare. Vidare att jämföra testernas EC50 medelvärde mot ISO 8692 angivet värde i precisions syfte och att utvärdera de båda räknemetoderna gällande precision och analystid. Kaliumdikromat användes för tillväxtinhibering vid toxicitetstest. EC50-resultaten visade ingen statistiskt signifikant skillnad mellan de båda instrumenten (p=0,47). Den akuta toxicitetsanalysens precision kunde bekräftas som giltig då båda mätmetoders EC50 medelvärden vid jämförelse med ISO 8692 värde befanns inom bestämda 95% konfidensintervallet. Vid jämförelse av studiens två mätningsmetoder, observerades större spridning kring medelvärdet i cellräknarens resultat, där tre EC50 värden hamnade utanför 95% CI. Däremot visade resultatet från flödescytometern mindre spridning och högre noggrannhet jämfört med cellräknarens. Studien visade att flödescytometer skulle kunna användas i framtida toxikologiska tester med encelliga alger, men det krävs flera upprepade försök för att bekräfta fördelar med analysen i flödescytometern.

Abstract [en]

Biological toxicity tests are performed by exposing a test organism to different concentrations of chemicals over a certain period of time. Results from acute studies are presented as EC50 (Effect Concentration, affecting 50% of the population). Tests used as a basis for risk assessments shall be performed in a quality acceptable way based on the International Organization for Standardization (ISO) and Good Laboratory Practice (GLP). The aim of this study was to study the toxic effect of potassium dichromate on unicellular green algae R. subcapitata, by counting cells with two different apparatus: flow cytometer and automatic cell counter. Additionally, to compare the EC50 mean values against the ISO 8692 value for control of test precision and to compare accuracy and analytical time of two methods. Potassium dichromate was used for growth inhibition in toxicity tests. The EC50 results showed no statistically significant difference between the two instruments (p = 0.47). The accuracy of acute toxicity analysis was confirmed as valid as both EC50 average measurement values compared to ISO 8692 value were found within the 95% confidence interval. When comparing the two methods of the study, greater spread was observed around the mean value in the cell count's results, where three EC50 values were outside 95% CI. The result of the flow cytometer had less spread and higher accuracy compared to the cell count. The study showed that flow cytometers could be used in future toxicological tests with algae, but several repeated tests are required to confirm the benefits of analysis with the flow cytometer.

Place, publisher, year, edition, pages
2018. , p. 30
Keywords [en]
Toxicity test, subcapitata, EC50, ISO 8692, potassium dichromate, flow cytometry, cell counter, growth inhibition
Keywords [sv]
Toxicitetstest, subcapitata, EC50, ISO 8692, kaliumdikromat, flödescytometer, cellräknare, tillväxtinhibering
National Category
Biological Sciences
Identifiers
URN: urn:nbn:se:hkr:diva-18257OAI: oai:DiVA.org:hkr-18257DiVA, id: diva2:1223003
Subject / course
Biomedical Laboratory Science
Educational program
Bachelor of Science in Biomedical Laboratory Science
Uppsok
Life Earth Science
Supervisors
Examiners
Available from: 2018-06-28 Created: 2018-06-24 Last updated: 2018-06-28Bibliographically approved

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