Natural Occurrence of Triploidy in a Wild Brown Bullhead

Cormier, Susan M.; Neiheisel, Timothy W.; Williams, Daniel E.; Tiersch, Terrence R.

doi:10.1577/1548-8659(1993)122<0390:nootia>2.3.co;2

Cited by 15 publications

(8 citation statements)

References 13 publications

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“…Triploid/diploid ratio of nuclear area by K r a s z n a i et aI. dimensions were also closer to findings of Cormier et al (1993) in image analysis of brown bullhead. It can be stated in accordance with Krasznai et al (1984) and Krasznai and Mar ian (1986) that erythrocyte nuclei of triploid wels differ significantly in mean size of nuclear major axis from those of diploids, even if maxima in distribution of this dimension in diploids can slightly overlap with minima in triploids, and they can be used for .-... .... identification oftriploidy.…”

Section: Discussionsupporting

confidence: 78%

A Model Approach to Distinguish Diploid and Triploid Fish by Means of Computer-assisted Image Analysis

Flajšhans¹

1997

Acta Vet. Brno

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F I aj s han sM.: A Model Approach to Distinguish Diploid and Triploid Fish by Means ofComputer-assisted Image Analysis. Acta vet. Brno 1997,66: 101-110. A model approach to distinguish diploid and triploid fish rapidly upon computer-assisted image analysis of erythrocyte nuclear dimensions in blood smear samples was exemplified on measurements of 13 diploid and 13 artificially-induced triploid wels (Silurus glanis). Analysis of variance test confirmed that nuclear area, perimeter, nuclear major axis and average nuclear size were the decisive dimensions for such approach (P < 0.000 I). The respecti ve values (mean ± SD) for diploids and triploids were registered as follows: nuclear area 11.09 ± 1.26 11m2 and 17.34 ±2.09 11m 2 ; nuclear perimeter 12.67 ± 0.80 11m and 16.24 ± 1.07 11m; nuclear major axis 4.62 ± 0.35 11m and 6.17 ± 0.47 11m and average nuclear size 4.0 I ± 0.22 11m and 5.12 ± 0.31 11m. This approach was found a rapid and precise alternative to other currently used methods of ploidy level identification in fish. Triploidy, image analysis, erythrocytes, nuclear dimensions, Silurus glanisThe differences in erythrocyte cellular and nuclear dimensions between diploid and triploid fish have been firstly registered by S w ar u p (1959). S e z a k i and K 0 bay a s i (1978) and L e m 0 i n e and S mit h (1980) have proposed a computation of cellular and nuclear area and volume from major and minor cellular and nuclear axes of diploid and polyploid fish. Later on, W 0 I t e r s et al. (1982) and Ben fey et al. (1984) proposed that identification of triploids could be based solely on measuring the erythrocyte nuclear major axis. To the best of my knowledge, computer -assisted image analysis of erythrocyte nuclei was used for identification of polyploid fish yet by Cor m i e r et al. (1993) only, for comparison of this technique as an alternative to flow cytometry and Coulter Counter for triploidy identification in brown bullhead, Ameiurus nebulosus.The goal of this paper was to use, describe and evaluate this innovative approach, the computer-assisted image analysis for discrimination of ploidy level based on erythrocyte nuclear dimensions. Diploid and triploid wels, Silurus glanis, was used as a model species. Materials and l\IethodsTriploid wels were produced at Anjou Fish Culture at Morannes. France. by heat shock as described by Lin h ar t and F I aj s han s (1995). Ploidy level of the fish was firstly assessed by flow cytometry at Angers University.Angers. France. according to Vi n del 0 v and C h r is ten sen (1990) and 13 diploid and 13 triploid yearlings were kept separately for this study. Blood was sampled from the caudal vessel into heparinized syringe and blood smears were prepared on microscopic slides according to Svobodova et al. (1986). then fixed with methanol. Slides were stained with 20% Giemsa stain in Sbrrensen's phosphate buffer (pH 6.8).Computer-assisted image analyses were carried out at the

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Section: Discussionsupporting

confidence: 78%

A Model Approach to Distinguish Diploid and Triploid Fish by Means of Computer-assisted Image Analysis

Flajšhans¹

1997

Acta Vet. Brno

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“…On the contrary, the NA was found to be the best ploidy level predictor distinguishing accurately among diploid, triploid and tetraploid erythrocytes without any further differences between live and fixed cells within a ploidy level. This is in agreement with the results of diploid and triploid erythrocytes by Cormier et al (1993) in brown bullhead, Ameiurus nebulosus ; Flajšhans (1997) and Linhart et al (2001) in European catfish, Silurus glanis ; Svobodová et al (1998) in tench; Abramenko et al (1998), Tóth et al (2005) and Vetešník et al (2006) in silver crucian carp; and with tetra‐, penta‐, hexa‐, hepta‐, octa‐ and dodeca‐ploid erythrocytes by Flajšhans and Vajcová (2000) andRáb et al (2004) in sturgeons. Similarly, the NP could be used to distinguish among diploid, triploid and tetraploid erythrocytes, as already proposed for triploids by Flajšhans (1997) and Svobodová et al (1998).…”

Section: Discussionsupporting

confidence: 89%

Image cytometric measurements of diploid, triploid and tetraploid fish erythrocytes in blood smears reflect the true dimensions of live cells

et al. 2010

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Comparative image cytometry of erythrocytes of diploid and triploid tench Tinca tinca L. and evolutionary tetraploid sterlet Acipenser ruthenus L. was performed on whole live unstained cells, live cells with stained nuclei and on stained fixed whole cells and their nuclei to test if erythrocyte measurements made from blood smears reflect the true dimensions of live cells. Nuclear area and perimeter were the best ploidy level predictors distinguishing accurately among live and fixed diploid, triploid and tetraploid cells, without significant differences between live and fixed cells within a ploidy level. Redundancy analysis revealed insignificant marginal effect of fixation (explained 2.3% of variation, F = 0.804), whereas the effect of ploidy level was highly significant (explained 50.6% of variation, F = 34.874). The erythrocyte measurements of diploid, triploid and tetraploid fish erythrocytes and their nuclei made from blood smears reflect the true dimensions of live cells, and the fixation procedure did not substantially affect their predictive value for ploidy level determination.

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“…Diploid and induced triploid tench, Tinca tinca (DNA content 2.04 and 3.10 pgDNA nucleus )1 , respectively) produced following the protocol of Flajsˇhans et al (2004) provided internal standards. Standard curve (IOD vs known DNA content) was generated and used primarily as a check that the stain was accurate across the range of the standards included (Cormier et al, 1993;Flajsˇhans, 1997;Svobodova´et al, 1998;Flajsˇhans andVajcova´, 2000, Linhart et al, 2001;Hardie et al, 2002). Feulgen image analysis densitometry following Hardie et al (2002) was conducted using a 3CCD Sony DXC-9100P camera coupled to an Olympus BX50 microscope (objective 100·), and the OLYMPUS MICRO-OLYMPUS MICRO-IMAGE IMAGE v. 4.0 image analysis software package (Olympus Corp., Tokyo, Japan) to measure integrated optical density (IOD) in erythrocyte nuclei, as well as the erythrocyte nuclear area (NA).…”

Section: Feulgen Image Analysismentioning

confidence: 99%

“…The RGB colour model software was set to the green channel only, with a maximal 190 pixel intensity; the light intensity calibration was set to Standard Optical Density. Standard curve (IOD vs known DNA content) was generated and used primarily as a check that the stain was accurate across the range of the standards included (Cormier et al, 1993;Flajsˇhans, 1997;Svobodova´et al, 1998;Flajsˇhans andVajcova´, 2000, Linhart et al, 2001;Hardie et al, 2002). A single primary standard (preferably chicken, DNA content 2.50 pgDNA nucleus )1 or tench DNA content 2.04 and 3.10 pgDNA nucleus )1 ) was used with confidence to calculate genome size.…”

Section: Feulgen Image Analysismentioning

confidence: 99%

Use of Feulgen image analysis densitometry to study the effect of genome size on nuclear size in polyploid sturgeons

Bytyutskyy

Srp

Flajšhans

2012

Journal of Applied Ichthyology

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Summary Feulgen image analysis densitometry (FIA) and image cytometry were used to study the relationship between the DNA content (pgDNA nucleus−1) and nuclear area (μm2) in blood smears of evolutionary tetraploid (4n) sterlet (Acipenser ruthenus) and stellate sturgeon (A. stellatus); evolutionary octaploid (8n) Siberian sturgeon (A. baerii) and Russian sturgeon (A. gueldenstaedtii); hexaploid (6n) and decaploid (10n) fish found within A. baerii stock; and A. baerii and A. gueldenstaedtii exhibiting dodecaploidy (12n). Standards used for FIA were blood smears of chicken (Gallus gallus domesticus; 2.5 pgDNA nucleus−1) and diploid and induced triploid tench, Tinca tinca (2.04 and 3.1 pgDNA nucleus−1, respectively). All ploidy levels were first verified by means of flow cytometry. Species of the same ploidy level, however differing in their DNA content, exhibited a similar mean erythrocyte nuclear area, as could be demonstrated on A. ruthenus and A. stellatus (19.27 and 19.79 μm2, respectively) with a respective mean DNA content of 3.72 and 4.68 pgDNA nucleus−1 and the same relationship was found for evolutionary octaploid (8n) A. baerii and A. gueldenstaedtii (29.87 and 30.09 μm2, respectively) with respective mean DNA content 8.29 and 7.87 pgDNA nucleus−1. The 0.19–0.32 pgDNA increments in DNA content of erythrocytes thus had no effect on their nuclear area. With increasing ploidy level, the DNA concentration (pgDNA per μm2 of erythrocyte nuclear area) was found not to increase linearly. The DNA in erythrocyte nuclei appeared to be more and more densely packed with an increase of the ploidy level (r = 0.98; R2 = 0.95).

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Natural Occurrence of Triploidy in a Wild Brown Bullhead

Cited by 15 publications

References 13 publications

A Model Approach to Distinguish Diploid and Triploid Fish by Means of Computer-assisted Image Analysis

A Model Approach to Distinguish Diploid and Triploid Fish by Means of Computer-assisted Image Analysis

Image cytometric measurements of diploid, triploid and tetraploid fish erythrocytes in blood smears reflect the true dimensions of live cells

Use of Feulgen image analysis densitometry to study the effect of genome size on nuclear size in polyploid sturgeons

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