A practical guide to unbiased quantitative morphological analyses of the gills of rainbow trout (Oncorhynchus mykiss) in ecotoxicological studies.
    
    
        
    
    
        
        PLoS ONE 15:e0243462 (2020)
    
    
    
      
      
	
	    Rainbow trout (Oncorhynchus mykiss) are frequently used as experimental animals in ecotoxicological studies, in which they are experimentally exposed to defined concentrations of test substances, such as heavy metals, pesticides, or pharmaceuticals. Following exposure to a broad variety of aquatic pollutants, early morphologically detectable toxic effects often manifest in alterations of the gills. Suitable methods for an accurate and unbiased quantitative characterization of the type and the extent of morphological gill alterations are therefore essential prerequisites for recognition, objective evaluation and comparison of the severity of gill lesions. The aim of the present guidelines is to provide practicable, standardized and detailed protocols for the application of unbiased quantitative stereological analyses of relevant morphological parameters of the gills of rainbow trout. These gill parameters inter alia include the total volume of the primary and secondary gill lamellae, the surface area of the secondary gill lamellae epithelium (i.e., the respiratory surface) and the thickness of the diffusion barrier. The featured protocols are adapted to fish of frequently used body size classes (300–2000 g). They include well-established, conventional sampling methods, probes and test systems for unbiased quantitative stereological analyses of light- and electron microscopic 2-D gill sections, as well as the application of modern 3-D light sheet fluorescence microscopy (LSFM) of optically cleared gill samples as an innovative, fast and efficient quantitative morphological analysis approach. The methods shown here provide a basis for standardized and representative state-of-the-art quantitative morphological analyses of trout gills, ensuring the unbiasedness and reproducibility, as well as the intra- and inter-study comparability of analyses results. Their broad implementation will therefore significantly contribute to the reliable identification of no observed effect concentration (NOEC) limits in ecotoxicological studies and, moreover, to limit the number of experimental animals by reduction of unnecessary repetition of experiments.
	
	
	    
	
       
      
	
	    
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        Publication type
        Article: Journal article
    
 
    
        Document type
        Scientific Article
    
 
    
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        Keywords
        Design-based Stereology; Surface-area; Fish Gill; Histopathological Alterations; Arbitrary Particles; Salmo-gairdneri; Pathological Research; Membrane Thickness; Prolonged Exposure; Brachydanio-rerio
    
 
    
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        Language
        english
    
 
    
        Publication Year
        2020
    
 
    
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        2020
    
 
    
    
        ISSN (print) / ISBN
        1932-6203
    
 
    
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	    Volume: 15,  
	    Issue: 12,  
	    Pages: ,  
	    Article Number: e0243462 
	    Supplement: ,  
	
    
 
    
        
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            Publisher
            Public Library of Science (PLoS)
        
 
        
            Publishing Place
            Lawrence, Kan.
        
 
	
        
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        Reviewing status
        Peer reviewed
    
 
     
    
        POF-Topic(s)
        30205 - Bioengineering and Digital Health
30505 - New Technologies for Biomedical Discoveries
    
 
    
        Research field(s)
        Enabling and Novel Technologies
    
 
    
        PSP Element(s)
        G-500390-001
A-630600-001
    
 
    
        Grants
        Hanns-Seidel-Stiftung e.V., German Federal Ministry of Education and Research (BMBF)
    
 
    
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        Erfassungsdatum
        2021-02-04