Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae
SUMMARY Antiparasitic pharmaceuticals are important groups of pharmaceuticals in veterinary medicine and are used in large quantities. Antiparasitic pharmaceuticals and their metabolites enter the environment via excretion in urine and/or faeces from treated animals and by run off to surface water...
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| Formato: | Second cycle, A2E |
| Lenguaje: | sueco Inglés |
| Publicado: |
2017
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| Materias: | |
| Acceso en línea: | https://stud.epsilon.slu.se/10148/ |
| _version_ | 1855571645596434432 |
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| author | Blomberg, Maria |
| author_browse | Blomberg, Maria |
| author_facet | Blomberg, Maria |
| author_sort | Blomberg, Maria |
| collection | Epsilon Archive for Student Projects |
| description | SUMMARY
Antiparasitic pharmaceuticals are important groups of pharmaceuticals in veterinary medicine and are used in large quantities. Antiparasitic pharmaceuticals and their metabolites enter the environment via excretion in urine and/or faeces from treated animals and by run off to surface water these compounds reach aquatic environments and thereby present a risk for aquatic organisms. The goal of this master thesis is to investigate if veterinary antiparasitic pharmaceuticals have any effects on the behaviour of zebrafish larvae. In addition, adding of a behavioural assay could increase sensitivity in the fish embryo toxicity test. There are some studies of toxicity of veterinary pharmaceuticals in aquatic animals, but they focus mainly on general toxic effects. Several of the antiparasitic pharmaceuticals has a neurotoxic mechanism of action, and may therefore affect behaviour of zebrafish larvae, which in turn can affect its responses to stimuli and the survival of the larvae in its natural environment.
In this study zebrafish larvae were exposed, from day 0 until day 6, to three different antiparasitic pharmaceuticals: doramectin, flumethrin and toltrazuril. An additional study was performed with doramectin, in order to study the effects of short time exposure. In the short term study the embryos were exposed to doramectin at day 6 for 1 h before behaviour recording. In the long time exposure experiments a behavioural assay was performed at day 6 using a video recording device (Viewpoint Zebrabox®) that recorded the swim activity of the larvae. Directly after the behavioural assay the larvae were assessed in stereo microscope for abnormalities and abnormal body posture. Only data from individuals that were considered as normal according to the macroscopic examination were included in the data set and used in the statistical analyses. Because of the large amount of data that were received from the recording device, four responses were chosen for statistical analysis: mean activity during the first period of darkness, mean activity during the first period of darkness, increase in activity during the first 10 seconds at the first period of darkness and difference between mean activity during the first and the last period of darkness.
Significant results were found for doramectin and flumethrin. The activity in the first five minutes dark period and the first five minutes light period were both significantly reduced in larvae exposed to the high concentration (0.58 mg/L) of doramectin compared with the control. Larvae exposed to the high concentration (0.11 mg/L) of flumethrin had a significant higher reduction in activity from the first to the last dark period as compared with the reduction in control larvae. The conclusion is that it is possible to detect altered behavioural response, in individuals that could not be classified as affected based on visual features. This finding supports that behavioural assays are potential tools to increase the sensitivity in toxicological studies in zebrafish. |
| format | Second cycle, A2E |
| id | RepoSLU10148 |
| institution | Swedish University of Agricultural Sciences |
| language | Swedish Inglés |
| publishDate | 2017 |
| publishDateSort | 2017 |
| record_format | eprints |
| spelling | RepoSLU101482017-04-24T12:04:31Z https://stud.epsilon.slu.se/10148/ Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae Blomberg, Maria Veterinary science and hygiene - General aspects SUMMARY Antiparasitic pharmaceuticals are important groups of pharmaceuticals in veterinary medicine and are used in large quantities. Antiparasitic pharmaceuticals and their metabolites enter the environment via excretion in urine and/or faeces from treated animals and by run off to surface water these compounds reach aquatic environments and thereby present a risk for aquatic organisms. The goal of this master thesis is to investigate if veterinary antiparasitic pharmaceuticals have any effects on the behaviour of zebrafish larvae. In addition, adding of a behavioural assay could increase sensitivity in the fish embryo toxicity test. There are some studies of toxicity of veterinary pharmaceuticals in aquatic animals, but they focus mainly on general toxic effects. Several of the antiparasitic pharmaceuticals has a neurotoxic mechanism of action, and may therefore affect behaviour of zebrafish larvae, which in turn can affect its responses to stimuli and the survival of the larvae in its natural environment. In this study zebrafish larvae were exposed, from day 0 until day 6, to three different antiparasitic pharmaceuticals: doramectin, flumethrin and toltrazuril. An additional study was performed with doramectin, in order to study the effects of short time exposure. In the short term study the embryos were exposed to doramectin at day 6 for 1 h before behaviour recording. In the long time exposure experiments a behavioural assay was performed at day 6 using a video recording device (Viewpoint Zebrabox®) that recorded the swim activity of the larvae. Directly after the behavioural assay the larvae were assessed in stereo microscope for abnormalities and abnormal body posture. Only data from individuals that were considered as normal according to the macroscopic examination were included in the data set and used in the statistical analyses. Because of the large amount of data that were received from the recording device, four responses were chosen for statistical analysis: mean activity during the first period of darkness, mean activity during the first period of darkness, increase in activity during the first 10 seconds at the first period of darkness and difference between mean activity during the first and the last period of darkness. Significant results were found for doramectin and flumethrin. The activity in the first five minutes dark period and the first five minutes light period were both significantly reduced in larvae exposed to the high concentration (0.58 mg/L) of doramectin compared with the control. Larvae exposed to the high concentration (0.11 mg/L) of flumethrin had a significant higher reduction in activity from the first to the last dark period as compared with the reduction in control larvae. The conclusion is that it is possible to detect altered behavioural response, in individuals that could not be classified as affected based on visual features. This finding supports that behavioural assays are potential tools to increase the sensitivity in toxicological studies in zebrafish. SAMMANFATTNING Antiparasitära läkemedel är en läkemedelsklass som används i stor mängd inom veterinärmedicinen och som sannolikt kommer ut i miljön via urin och faeces från behandlade djur. Via bland annat avrinning från åkrar där gödsel och urin från djuren spridits så kan dessa substanser, och dess eventuella metaboliter, nå vattendrag och därmed utgöra en risk för vattenlevande organismer. Syftet med studien är att studera hur zebrafiskyngels beteende påverkas av tre olika läkemedels substanser ur denna läkemedelsklass. Ett annat syfte var även att undersöka om beteendestudier kan öka känsligheten i toxiska tester. Det finns en del data gällande generella toxiska effekter av veterinärmedicinska läkemedel på vattenlevande organismer, däremot få studier om hur de påverkar beteendet hos dessa djur. Flera antiparasitära läkemedel är neurotoxiska och antas därigenom kunna påverka beteende, vilket i sin tur antas kunna påverka dessa djurs respons på olika stimuli och därigenom deras överlevnad i naturen. I denna studie exponerades zebrafiskembryon från dag 0 till dag 6 för tre olika läkemedelssubstanser, doramektin, flumetrin och toltrazuril enligt en standardiserad metod. Det gjordes även ett försök med korttidsexponering där embryona exponerades för doramektin dag 6, under en timme innan avläsning. I långtidsexponeringarna utfördes en beteendeanalys dag 6 avseende påverkan på aktivitetsnivån (simaktiviteten) med hjälp av ett videoanalyssystem (Viewpoint Zebrabox®). Direkt efter videoanalysen gjordes en visuell bedömning av embryona avseende fysiologiska och morfologiska avvikelser samt avvikelser i kroppspositionering. Endast data från embryon som inte uppvisade några förändringar inkluderades i de data som analyserades statistiskt. Ur den stora mängd data som erhölls valdes fyra responser ut för statistisk analys: aktivitet under de 10 första sekunderna under första mörkerperioden, medelaktivitet under första mörkerperioden, medelaktivitet under första ljusperioden, samt skillnad i medelaktivitet mellan första och sista mörkerperioden. Signifikant resultat erhölls för högsta koncentrationen av flumetrin och doramektin. Flumetrin i koncentrationen 0,11 mg/L gav en signifikant sänkning i medelaktivitet mellan första och sista mörkerperioden hos de exponerade ynglen jämfört med kontrollen. Yngel som exponerats för doramektin i koncentrationen 0,58 mg/L hade lägre aktivitet både under första mörkerperioden och under första ljusperioden. Det var således möjligt att med metoden detektera beteendeförändringar hos yngel som vid en visuell bedömning klassificerades som normala och detta talar för att beteendeanalys kan vara användbart för att öka känsligheten i toxikologiska studier på zebrafisk. 2017-04-12 Second cycle, A2E NonPeerReviewed application/pdf sv https://stud.epsilon.slu.se/10148/1/blomberg_m_20170424.pdf Blomberg, Maria, 2017. Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae. Second cycle, A2E. Uppsala: (VH) > Dept. of Biomedical Sciences and Veterinary Public Health (until 231231) <https://stud.epsilon.slu.se/view/divisions/OID-713.html> urn:nbn:se:slu:epsilon-s-6360 eng |
| spellingShingle | Veterinary science and hygiene - General aspects Blomberg, Maria Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| title | Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| title_full | Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| title_fullStr | Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| title_full_unstemmed | Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| title_short | Veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| title_sort | veterinary antiparasitic pharmaceuticals – effects on behaviour in fish larvae |
| topic | Veterinary science and hygiene - General aspects |
| url | https://stud.epsilon.slu.se/10148/ https://stud.epsilon.slu.se/10148/ |