GEOMETRIC MORPHOMETRIC ANALYSIS OF SILVER CARP LARVAE AFTER EXPOSURE TO TDCIPP
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摘要: 为了解磷酸三(1,3-二氯-2-丙基)酯(TDCIPP)对鲢仔鱼生长抑制性在外部形态上的表现性状, 研究基于几何形态测量学方法对0.05、0.5、5和50 μg/L共4个浓度组与对照组进行组间形态性状差异比较分析。利用PLYMPLUS系统获取鲢仔鱼样本图像信息并测量体长, 利用万分电子天平称量体重, 使用TPS系列软件提取坐标点数据, 并通过Morpho J软件完成主成分分析、典型变量分析及结果可视化。除0.05 μg/L浓度组外, 其他浓度组鲢仔鱼的体长、体重均显著低于对照组, 表明TDCIPP对鲢仔鱼的生长发育具有抑制效应。主成分分析和典型变量分析结果显示, 第一主成分(PC1)和第二主成分(PC2)共占总体变量的62.15%(分别为47.64%和14.51%); 第一典型变量(CV1)和第二典型变量(CV2)共占总体变量的79.48%(分别为54.55%和24.93%), 满足用于鲢仔鱼形态分析的要求。网格轮廓分析结果显示, 各浓度组鲢仔鱼平均形态均与对照组间存在显著差异(P＜0.05), 并且主要表现为头部、躯干纵轴和尾部发育迟缓。Abstract: Tris (1,3-dichloro-2-propyl) phosphate (TDCIPP), widely used as a kind of organophosphorus flame retardant, has been detected in the Yangtze River water environments. Many toxicological assessments have shown that TDCIPP could change morphology of fish. Silver carp (Hypophthalmichthys molitrix) lives in the Yangtze River for its entire life story, however, the effects of TDCIPP on silver carp is unclear. In order to clarify the main morphological characters of growth inhibition of silver carp larvae caused by TDCIPP, the present study analyzed the morphological traits between four environmentally relevant concentrations (0.05, 0.5, 5 and 50 μg/L) and the control group by geometric morphometric analysis. After the image information of larvae simple was obtained, the body length and body weight were measured. Then, digitization of landmarks was carried out with the TPS series software. Finally, principal component analysis (PCA), canonical variates analysis (CVA) and results visualization were carried out with Morpho J software. The body length and body weight of silver carp larvae decreased significantly under exposure to 0.5, 5 and 50 μg/L of TDCIPP compared with the control group, but no effects were observed in 0.05 μg/L. This indicated that environmentally relevant concentrations of TDCIPP induced growth inhibition in silver carp larvae. The results of PCA and CVA indicated that the first principal component (PC1) and the second principal component (PC2) together accounted for 62.15% of the overall variables (47.36% and 14.51%, respectively). The first canonical variates (CV1) and the second canonical variates (CV2) together accounted for 79.48% (54.55% and 24.93%, respectively), which satisfied the requirement of morphological analysis of silver carp larvae. The results of grid profile analysis indicated that the average morphology of silver carp larvae in different concentrations was significantly different with the control group (P＜0.05), which identified by the growth retardation of the head, longitudinal axis of body and tail. As a conclusion, TDCIPP could induce the growth retardation of head, longitudinal axis of body and tail in silver carp larvae. Therefore, attentions should be paid to the environmental concentrations of TDCIPP in the Yangtze River Basin, and the ecological risk of TDCIPP to the replenishment of silver carp population resources should be assessed.
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Keywords:
- Geometric morphometric analysis /
- Morpho J /
- TDCIPP /
- Growth inhibition /
- Silver carp larvae
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图 2 鲢仔鱼5个实验组坐标点的普氏叠印图
实心点表示鲢仔鱼的平均形态
Figure 2. The Procrustes diagram of five experimental groups of silver carp larvae
Solid points indicate the average morphology of silver carp larvae
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图 3 不同实验组鲢仔鱼体长、体重的小提琴图
黑色实线代表平均值; 虚线代表四分位数; **表示P＜0.01; ****表示P＜0.001；ns表示P > 0.05
Figure 3. Violin plot of body length and body weight of silver carp larvae with different experimental groups
Solid black lines indicate the mean; dotted lines indicate the quartiles; ** indicate P＜0.01; **** indicate P＜0.001；ns indicate P > 0.05
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图 4 鲢仔鱼形态第一主成分和第二主成分的散点图及网格轮廓图
A和B分别表示PC1轴的值为0.1和–0.1时的鲢仔鱼平均形态; C和D分别表示PC2轴的值为–0.12和0.12时的鲢仔鱼平均形态; E. 圆圈表示不同实验组鲢仔鱼形态均值的95%置信椭圆
Figure 4. The scatter plots of PC1 and PC2 for morphology and outline variation of silver carp larvae
Mean shapes of silver carp larvae are shown for sores of 0.1 (A) and –0.1 (B) in PC1. Mean shapes of silver carp larvae are shown for sores of –0.12 (C) and 0.12 (D) in PC2; E. the circle represents 95% confidence ellipses of the mean shapes of silver carp in different experimental groups
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图 5 鲢仔鱼形态第一典型变量和第二典型变量的散点图及网格轮廓图
A和B分别表示CV1轴的值为5.0和–4.0时的鲢仔鱼平均形态; C和D分别表示CV2轴的值为–7.0和4.0时的鲢仔鱼平均形态; E. 圆圈表示不同实验组鲢仔鱼形态均值的95%置信椭圆
Figure 5. The scatter plots of CV1 andCV2 for morphology and outline variation of silver carp larvae
Mean shapes of silver carp larvae are shown for sores of 5.0 (A) and –4.0 (B) in CV1. Mean shapes of silver carp larvae are shown for sores of –7.0 (C) and 4.0 (D) in CV2; E. the circle represents 95% confidence ellipses of the mean shapes of silver carp in different experimental groups
表 1 坐标点类型与定义
Table 1 The landmark type and definition
坐标点类型
Landmark type定义
DefinitionⅠ型坐标点 3 背鳍起点 Origin of dorsal fin 4 背鳍基部末端 Posterior end of dorsal fin base 5 尾鳍基部上端 Upper insertion of caudal fin base 7 尾鳍基部下端 Lower insertion of caudal fin base 8 臀鳍基部末端 Posterior end of anal fin base 9 臀鳍起点 Origin of anal fin 10 腹鳍起点 Origin of pelvic fin Ⅱ型坐标点 2 额骨 Frontal bone 6 尾柄末端 Distal tip of caudal peduncle 11 前鳃盖腹侧起点 Origin of the preopercle on the ventral 12 眼前缘 The anterior margin of the eye 13 眼后缘 The posterior margin of the eye Ⅲ型坐标点 1 吻端 Tip of snout 
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表 2 鲢仔鱼形态主成分分析
Table 2 Principal component analysis of morphology in silver carp larvae
主成分
Principal
component特征值
Eigenvalue占总体变异
Variance (%)累计总体变异
Cumulative
variance (%)PC1 0.00138356 47.64 47.64 PC2 0.00042130 14.51 62.15 PC3 0.00031310 10.78 72.93 PC4 0.00013657 4.70 77.63 PC5 0.00011275 3.88 81.51 PC6 0.00010520 3.62 85.13
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表 3 鲢仔鱼形态典型变量分析
Table 3 Canonical variate analysis of morphology in silver carp larvae
典型变量
Canonical
variate特征值
Eigenvalue占总体变异
Variance (%)累计总体变异
Cumulative
variance (%)CV1 0.40673144 54.55 54.55 CV2 0.18592402 24.93 79.48 CV3 0.08441425 11.32 90.80 CV4 0.06859854 9.20 100.00
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表 4 不同实验组鲢仔鱼13个坐标点矩阵基于典型变量分析计算的马氏距离和普氏距离
Table 4 Mahalanobis and Procrustes distance computed from a canonical variates analysis of a matrix of 13 landmark of silver carp larvae
对照组
Control0.05 μg/L 0.5 μg/L 5 μg/L 50 μg/L 马氏距离Mahalanobis distance 对照组Control 0.0004 <0.0001 <0.0001 <0.0001 0.05 μg/L 1.0538 <0.0001 <0.0001 <0.0001 0.5 μg/L 1.6705 1.1827 <0.0001 <0.0001 5 μg/L 1.3086 1.1278 1.1546 50 μg/L 1.8497 1.5651 1.3084 1.1161 普氏距离Procrustes distance 对照组Control 0.0299 <0.0001 0.0008 <0.0001 0.05 μg/L 0.0138 0.0269 0.4196 0.0006 0.5 μg/L 0.0262 0.0137 0.0990 0.0385 5 μg/L 0.0187 0.0076 0.0107 0.0036 50 μg/L 0.0322 0.0205 0.0132 0.0169 注: 上三角表示P值, 下三角表示不同群体间的距离; 种群间距离显著性的P值是通过排列试验(10000个重复)计算出来的Note: Upper triangle are P-values, lower triangle are distances between populations; P-values for the significance of the interpopulation distances were computed using permutation tests (10000 replications)
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