Local connected fractal dimension analysis in gill of fish experimentally exposed to toxicants.

An operator-neutral method was implemented to objectively assess European seabass, Dicentrarchus labrax (Linnaeus, 1758) gill pathology after experimental exposure to cadmium (Cd) and terbuthylazine (TBA) for 24 and 48h. An algorithm-derived local connected fractal dimension (LCFD) frequency measure was used in this comparative analysis. Canonical variates (CVA) and linear discriminant analysis (LDA) were used to evaluate the discrimination power of the method among exposure classes (unexposed, Cd exposed, TBA exposed). Misclassification, sensitivity and specificity, both with original and cross-validated cases, were determined. LCFDs frequencies enhanced the differences among classes which were visually selected after their means, respective variances and the differences between Cd and TBA exposed means, with respect to unexposed mean, were analyzed by scatter plots. Selected frequencies were then scanned by means of LDA, stepwise analysis, and Mahalanobis distance to detect the most discriminative frequencies out of ten originally selected. Discrimination resulted in 91.7% of cross-validated cases correctly classified (22 out of 24 total cases), with sensitivity and specificity, respectively, of 95.5% (1 false negative with respect to 21 really positive cases) and 75% (1 false positive with respect to 3 really negative cases). CVA with convex hull polygons ensured prompt, visually intuitive discrimination among exposure classes and graphically supported the false positive case. The combined use of semithin sections, which enhanced the visual evaluation of the overall lamellar structure; of LCFD analysis, which objectively detected local variation in complexity, without the possible bias connected to human personnel; and of CVA/LDA, could be an objective, sensitive and specific approach to study fish gill lamellar pathology. Furthermore this approach enabled discrimination with sufficient confidence between exposure classes or pathological states and avoided misdiagnosis.

[1]  E. Wolin,et al.  Texture analysis of medical images in radiotherapy , 2016 .

[2]  E Le Rumeur,et al.  MRI texture analysis on texture test objects, normal brain and intracranial tumors. , 2003, Magnetic resonance imaging.

[3]  K. Choe,et al.  The multifunctional fish gill: dominant site of gas exchange, osmoregulation, acid-base regulation, and excretion of nitrogenous waste. , 2005, Physiological reviews.

[4]  Benoit B. Mandelbrot,et al.  Fractal Geometry of Nature , 1984 .

[5]  W. B. Marks,et al.  Fractal methods and results in cellular morphology — dimensions, lacunarity and multifractals , 1996, Journal of Neuroscience Methods.

[6]  N. Kenkel,et al.  Fractals in the Biological Sciences , 1996 .

[7]  Doaa Mahmoud-Ghoneim,et al.  Texture analysis of magnetic resonance images of rat muscles during atrophy and regeneration. , 2006, Magnetic resonance imaging.

[8]  J. Mallatt.  Fish Gill Structural Changes Induced by Toxicants and Other Irritants: A Statistical Review , 1985 .

[9]  K. Willett,et al.  Gill Histopathologies Following Exposure to Nanosilver or Silver Nitrate , 2015, Journal of toxicology and environmental health. Part A.

[10]  Arvydas Laurinavicius,et al.  Digital Image Analysis in Pathology: Benefits and Obligation , 2011, Analytical cellular pathology.

[11]  A. Abdel-Moneim,et al.  Gill oxidative stress and histopathological biomarkers of pollution impacts in Nile tilapia from Lake Mariut and Lake Edku, Egypt. , 2012, Journal of aquatic animal health.

[12]  T. Wahli,et al.  Histopathology in fish: proposal for a protocol to assess aquatic pollution , 1999 .

[13]  G. Landini,et al.  Local connected fractal dimensions and lacunarity analyses of 60 degrees fluorescein angiograms. , 1995, Investigative ophthalmology & visual science.

[14]  Michal Strzelecki,et al.  A software tool for automatic classification and segmentation of 2D/3D medical images , 2013 .

[15]  G Landini,et al.  Fractals in microscopy , 2011, Journal of microscopy.

[16]  Joseph S. Meyer,et al.  Structural changes in gills of Lost River suckers exposed to elevated pH and ammonia concentrations. , 2003, Comparative biochemistry and physiology. Toxicology & pharmacology : CBP.

[17]  E. Agamy.  Sub chronic exposure to crude oil, dispersed oil and dispersant induces histopathological alterations in the gills of the juvenile rabbit fish (Siganus canaliculatus). , 2013, Ecotoxicology and environmental safety.

[18]  C. Brauner,et al.  Ontogeny and paleophysiology of the gill: New insights from larval and air-breathing fish , 2012, Respiratory Physiology & Neurobiology.

[19]  David H. Evans,et al.  The fish gill: site of action and model for toxic effects of environmental pollutants. , 1987, Environmental Health Perspectives.

[20]  B. Dezfuli,et al.  The use of fractal dimension and lacunarity in the characterization of mast cell degranulation in rainbow trout (Onchorhynchus mykiss) , 2014, Journal of microscopy.

[21]  M. Álvarez-Dolado,et al.  Cell fusion and tissue regeneration. , 2011, Advances in experimental medicine and biology.

[22]  L. Seuront.  Fractals and Multifractals in Ecology and Aquatic Science , 2009 .

[23]  Ketil Hylland,et al.  Quantitative changes in metallothionein expression in target cell-types in the gills of turbot (Scophthalmus maximus) exposed to Cd, Cu, Zn and after a depuration treatment. , 2006, Aquatic Toxicology.

[24]  J. Mancera,et al.  A stereological study of copper toxicity in gills of Oreochromis niloticus. , 2009, Ecotoxicology and environmental safety.

[25]  G. M. Hughes,et al.  A morphometric study of the effects of tributyltin compounds on the gills of the mummichog, Fundulus heteroclitus , 1989 .

[26]  A. Sales,et al.  Can fish gill anomalies be used to assess water quality in freshwater Neotropical systems? , 2012, Environmental Monitoring and Assessment.

[27]  Shabanam S. Tamboli,et al.  Histopathological Image Analysis for Breast Cancer Diagnosis : A Review , .

[28]  A. A. Nascimento,et al.  Fish Gills Alterations as Potential Biomarkers of Environmental Quality in a Eutrophized Tropical River in South‐Eastern Brazil , 2012, Anatomia, histologia, embryologia.

[29]  Jonathan M. Wilson,et al.  Fish gill morphology: inside out. , 2002, The Journal of experimental zoology.

[30]  James Diamond,et al.  The use of morphological characteristics and texture analysis in the identification of tissue composition in prostatic neoplasia. , 2004, Human pathology.

[32]  B. Dezfuli,et al.  European sea bass gill pathology after exposure to cadmium and terbuthylazine: expert versus fractal analysis , 2016, Journal of microscopy.

[33]  Constantinos G Loukas,et al.  A survey on histological image analysis-based assessment of three major biological factors influencing radiotherapy: proliferation, hypoxia and vasculature , 2004, Comput. Methods Programs Biomed..

[34]  R. Triebskorn,et al.  Ultrastructural changes in fish gills as biomarker to assess small stream pollution. , 1998, Tissue & cell.

[35]  M. Manera.  The Use of Texture Analysis in the Morpho-Functional Characterization of Mast Cell Degranulation in Rainbow Trout (Onchorhynchus mykiss) , 2013, Microscopy and Microanalysis.

[36]  Markus J. Tamás,et al.  Heavy Metals and Metalloids As a Cause for Protein Misfolding and Aggregation , 2014, Biomolecules.

[37]  Klaus Weber,et al.  Nonlesions, Misdiagnoses, Missed Diagnoses, and Other Interpretive Challenges in Fish Histopathology Studies , 2015, Toxicologic pathology.

[38]  B. Dezfuli,et al.  Effects of experimental terbuthylazine exposure on the cells of Dicentrarchus labrax (L.). , 2006, Chemosphere.

[39]  B. Dezfuli,et al.  Cellular alterations in different organs of European sea bass Dicentrarchus labrax (L.) exposed to cadmium. , 2007, Chemosphere.

[40]  C. Wood,et al.  The physiological consequences of exposure to chronic, sublethal waterborne nickel in rainbow trout (Oncorhynchus mykiss): exercise vs resting physiology , 2004, Journal of Experimental Biology.

[41]  E. Weibel,et al.  Fractals in Biology and Medicine , 1994 .

[42]  Artur Klepaczko,et al.  Texture and color based image segmentation and pathology detection in capsule endoscopy videos , 2014, Comput. Methods Programs Biomed..

[43]  Joel H. Saltz,et al.  Histopathological Image Analysis Using Model-Based Intermediate Representations and Color Texture: Follicular Lymphoma Grading , 2009, J. Signal Process. Syst..

[44]  Shaimaa Al-Janabi,et al.  Digital pathology: current status and future perspectives , 2012, Histopathology.

[45]  A. Madabhushi.  Digital pathology image analysis: opportunities and challenges. , 2009, Imaging in medicine.

[46]  S. Raisuddin,et al.  Effects of exposure to multiple trace metals on biochemical, histological and ultrastructural features of gills of a freshwater fish, Channa punctata Bloch. , 2008, Chemico-biological interactions.