Morphometrical study of physical growth in infant cynomolgus monkeys using multivariate analysis

We describe the relative growth in infant cynomolgus monkeys (Macaca fascicularis). The multivariate analyses using the principal component analysis and multivariate allometric analysis were applied to biometrical data of laboratory-bred cynomolgus monkeys aged from birth to 12 weeks. All the animals had been bred and reared under uniform environmental conditions at Tsukuba Primate Center for Medical Science, N. I. H., Japan. As the result of the principal component analysis, the cumulative contribution ratio of the 1st and 2nd principal components (PC1 and PC2) accounted for 88.2% and 85.7% in males and females, respectively. This indicates that the morphologic traits (15 traits) grow normally in size, but the analysis did not detect any differences in growth rate among the traits. So, the multivariate allometric analysis was applied to the same data. The traits concerning the face and truck showed relatively high growth rates compared to head breadth, whereas the limbs and arms grew slowly.     

Chimeric heterosis in mandibular size in mice

Morphometrical observations were carried out on the mandibles of chimeras made from the embryos of C57BL/6 and BALB/c mice to compare with the two strains and their reciprocal F1 crosses. The results of the principal component analysis indicate that the first principal component (PC1) and the second principal component (PC2) extracted might be acceptable as size and shape factors, respectively. Variations of both PC1 and PC2 were generally larger in the chimeras than in the two component strains and their F1 crosses. The mean PC1 value of the chimeras was larger than that of the two component inbred strains, and it was similar to that of F1 crosses, or slightly larger. The overall size of the mandible represented by PC1 tended to be larger in the chimeras consisting of two component cells that were approximately equivalent than in those that shifted to either cell population. The above trend was observed in both sexes. These results indicate that chimeric heterosis due to the interaction between genetically different cells (C57BL/6 and BALB/c) has some relation to mandible size. The mean PC2 value, which was accepted as shape factor, was intermediate between the two inbred strains. The mandible size (PC1) and shape (PC2) were bilaterally symmetrical, except for the shape in the female chimeras and in (C57BL/6 x BALB/c)F1.     

The ontogeny of sexual dimorphism in the cranium of Bornean orang-utans (Pongo pygmaeus pygmaeus) as detected by principal-components analysis

The ontogeny of cranial sexual dimorphism in the Bornean orang-utan (Pongo pygmaeus pygmaeus) is examined by means of principal-components analysis (PCA). Normalized first components are called allometry vectors or vectors of relative growth and show that sexual dimorphism is present at all stages of growth. Two patterns of sexual dimorphism are present: (1) sexual differences at age groups 2 and 3 are the result primarily of differences in principal component II scores, reflecting mainly shape-related differences, and (2) age groups 5, 6, and 7 show a trend of stronger size-related shape differences with increasing age in the allometry vector along with decreasing differences in principal component II scores, reflecting an increase in size-related shape differences between the sexes. Age group 4 shows a combination of both patterns. Our results support Shea's hypothesis (1985a) that when using multigroup PCAs in closely related taxa, the allometry vector will generally estimate the shape variation resulting from the extension of common growth allometry patterns (ontogenetic scaling). The second and subsequent components summarize shape variation from slope and intercept differences between the groups, provided that ontogenetic scaling is not solely responsible for all the shape differences present. Subanalyses of those dimensions previously found to show ontogenetic scaling and acceleration follow this pattern well. The total sample provides a pattern whereby ontogenetically scaled dimensions possess a stronger influence over accelerated dimensions but still generally follow Shea's hypothesis. Finally, variously derived coefficients provided several interesting findings: (1) strong evidence was found against multivariate isometry for both the pooled and the separate samples, (2) multivariate allometric coefficients for both sexes follow the general growth pattern of negative scaling in neurocranial dimensions and positive scaling in the viscerocranium, and (3) multivariate slopes have a very high correlation with bivariate slopes relative to the same independent X variable, thereby lending further support to Jolicoeur's (1963a, b) allometry generalization.     

Heterochrony and geometric morphometrics: a comparison of cranial growth in Pan paniscus versus Pan troglodytes

Heterochrony, the classic framework in which to study ontogeny and phylogeny, in essence relies on a univariate concept of shape. Though principal component (PC) plots of multivariate shape data seem to resemble classical bivariate allometric plots, the language of heterochrony cannot be translated directly into general multivariate methodology. We simulate idealized multivariate ontogenetic trajectories and explore their appearance in PC plots of shape space and size-shape space. Only if the trajectories of two related species lie along exactly the same path in shape space can the classic terminology of heterochrony apply and pure dissociation of size change against shape change be detected. Regional heterochrony--the variation of apparent heterochrony by region--implies a dissociation of local growth fields and cannot be identified in an overall PC analysis. We exemplify a geometric morphometric approach to these issues using adult and subadult crania of 48 Pan paniscus and 47 Pan troglodytes specimens. On each specimen, we digitized 47 landmarks and 144 semilandmarks on facial curves and the external neurocranial surface. We reject the hypothesis of global heterochrony in the cranium of Pan as well as regional heterochrony for the lower face, the upper face, and the neurocranium.     

Principles and methods of geometric morphometrics

The basic concepts, notions and methods of geometric morphometrics (GM) are considered. This approach implies multivariate analysis of landmark coordinates located following certain rules on the surface of a morphological object. The aim of GM is to reveal differences between morphological objects by their shapes as such, the "size factor" being excluded. The GM is based on the concept of Kendall's space (KS) defined as a hypersphere with points distributed on its surface. These points are the shapes defined as aligned landmark configurations. KS is a non-Euclidian space, its metrics called Procrustes is defined by landmark configuration of a reference shape relative to which other shapes are aligned and compared. The differences among shapes are measured as Procrustes distances between respective points. For the linear methods of multivariate statistics to be applied to comparison of shapes, the respective points are projected onto the tangent plane (tangent space), the tangent point being defined by the reference. There are two principal methods of shape comparisons in GM: the Procrustes superimposition (a version of the least squares analysis) and thin-plate spline analysis. In the first case, Procrustes residuals are the outcome shape variables which remain after isometric alignment of the shapes being compared. Their summation over all landmarks yields Procrustes distances among these shapes. The Procrustes distances can be used in multivariate analyses just as the Euclidian distances. In the second case, the shapes are fitted to the references by stretching/compressing and shearing until complete identity of their landmark configurations. Eigenvectors of resulting bending energy matrix are defined as new shape variables, principal warps which yield another shape space with the origin defined by the reference. Projections of the shapes being compared onto principal warps yield partial warps, and their covariance matrix decomposition into eigenvectors yields relative warps which are similar to principal components (in particular, they are mutually orthogonal). Both partial and relative warps can be used in many multivariate statistic analyses as quantitative shape variables. Results of thin-plate spline analysis can be represented graphically by transformation grid which displays type, amount and localization of the shape differences. Basis rules of sample composition and landmark positioning to be used in GM are considered. At present, rigid (with minimal degrees of freedom) 2D morphological objects are most suitable for GM applications. It is important to recognize three type of real landmarks, and additionally semi-landmarks and "virtual" landmarks. Some procedures of thin-plate spline analysis are considered exemplified by some study cases, as well as applications of some standard multivariate methods to GM results. They make it possible to evaluate correlation between different shapes, as well as between a shape and some non-shape variables (linear measurements etc); to evaluate the differences among organisms by shape of a morphological structure; to identify landmarks which most accounted for both correlation and differences between the shapes. An annotated list of most popular softwares for GM is provided.     

Morphological characterization of ejaculated cynomolgus monkey (Macaca fascicularis) sperm

The aim of this study was to give reference values for the frequency of morphological sperm abnormalities present in the semen from nonexperimental cynomolgus monkeys as well as for the dimensions of sperm heads. Spermatozoa from the liquid portion of electroejaculates from 14 cynomolgus monkeys were air‐dried as smears, fixed, and stained with Harris's Haematoxylin and subjected to visual analysis of morphology and computer‐aided analysis of ten morphometric variables. The majority (83%) of sperm were morphologically normal. Tail defects were the most common (11%), and showed the highest variation between individuals, the values ranging between 4 and 23%. Head abnormalities consisted of large, tapering, and amorphous forms but were not frequent (0.4%), the values ranging between 0 and 1.3%. Midpiece imperfections were found in all the individuals; the mean percentage was 5%, and the range varied between 3 and 9%. Tail plus midpiece was the only multiple abnormality observed, with a mean value of 1.5% and a range between 0 and 8%. The majority of these double defects consisted of a coiled tail together with a coiled midpiece. Mean values for the morphometric parameters characterizing sperm heads were as follows: area 17.2 μm², perimeter 15.2 μm, length 5.8 μm, width 4.0 μm, L/W ratio 1.5, gray‐level 98, ellipticity 0.2, first shape factor 0.9, second shape factor 1.4, and third shape factor 1.1. Overall coefficients of variation for the majority of parameters were below 7%, showing the great homogeneity in the dimensions of cynomolgus sperm heads. Most useful parameters for sperm characterization, according to their low variability, were perimeter, length, width, L/W ratio, and shape factors. Differences in these parameters were, however, observed between monkeys. Am. J. Primatol. 47:105–115, 1999. © 1999 Wiley‐Liss, Inc.     

Bivariate and multivariate growth allometry: statistical and biological considerations

Bivariate and multivariate analyses of 55 cranial dimensions were completed on ontogenetic series of Pygmy chimpanzees ( Pan paniscus ), Common chimpanzees ( Pan troglodytes ), and gorillas ( Gorilla gorilla ). Subanalyses described here were specifically designed to compare and crosscheck quantitative assessments of relative growth as determined using the bivariate and multivariate (principal components analysis—PCA) approaches. Results indicate a strong concordance between the bivariate and multivariate patterns, empirically supporting the claim that PCA provides an effective multivariate approach to analysing growth allometry. Comparison of bivariate and multivariate results also suggests that in multi-group PCA, the first component summarizes shape variation resulting from the sharing and extension of common patterns of growth allometry, while the second and subsequent components summarize shape variation resulting from divergent growth trajectories, reflected in bivariate comparisons as either vertical shifts and/or slope differences. Examination of non-normalized first component loadings, plus a comparison with estimates of logarithmic growth in the cranial dimensions, reveals that the non-normalized loadings are proportional to coefficients of specific growth. This finding further links the bivariate and multivariate approaches, grounding both in Huxley's theoretical notions of multiplicative and relative growth.     

基于几何形态测量学的磷酸三(1,3-二氯-2-丙基)酯对鲢仔鱼形态影响研究

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50μg/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0.05μg/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。     

Evolution of adaptation through allometric shifts in a marine snail

Variation in ontogenetic development among individuals may be a major contributor to morphological variation within species. Evolution of different growth trajectories might, for example, evolve as a response to varying ecological contexts of individuals living in different environments, or by life-stage or gender differences. The intertidal periwinkle Littorina saxatilis is strongly polymorphic in shell shape. We compared ontogenetic trajectories between life stages, local populations, and sexes to understand how different morphological end points are reached during ontogeny and what might cause these differences. Applying landmark-based geometric morphometrics, we captured shell shape variation for four Swedish populations of this species. We also derived a method to visualize ontogenetic trajectories described by the relationship of size to the multivariate shape space. We found that growth trajectories differed between individuals living in different habitats, as well as between sexes and maturity stages. Males living on rocky cliffs grew isometrically throughout life, whereas females from the same habitat switched from isometric growth as juveniles to allometric growth as adults. In contrast, males and females living on boulders grew allometrically as juveniles but changed to isometric growth at sexual maturity. Thus, in this species, ontogenetic growth seems influenced by habitat-associated selection as well as by gender and age-specific selection. These differing selection regimes result in ontogenetic shifts in allometry in three of the four groups examined.     

珠江卷口鱼不同地理种群的形态变异

运用传统生物形态学和框架分析方法,对珠江水系中栖息于柳江、浔江、西江的卷口鱼(Ptychidio jordani Myers)的形态变异进行了分析。根据高背、平背体型分析了体长/体高比例与地理、性别和年龄的相关性。结果显示:体长/体高比例与年龄呈显著负相关,随着年龄增长,卷口鱼逐渐趋向于高背型;三个江段卷口鱼可数性状数据卡方检验没有显著差异;三个江段卷口鱼可量性状数据和框架数据聚类分析、判别分析和主成分分析显示三个江段的卷口鱼形态差异明显,有一定程度的分化;聚类和主成分分析显示柳江群体变异最大,而西江卷口鱼则分化成两个群体,其中一个与浔江群体关系较近。判别分析构建的判别公式可以区分三个江段四个群体的卷口鱼,判别正确率达到99.2%。不同江段卷口鱼的差异表现在整个鱼体上,不能用单独的一或二个参数进行鉴别。     

Multivariate morphometries and sexual dimorphism in the orb-web spider Metellina segmentata (Clerck, 1757) (Araneae, Metidae)

Sexual dimorphism in body size and leg length was investigated in a common orb-weaving spider of Ireland and northern Europe, Metellina segmentata (Clerck, 1757) (Araneae, Metidae). Univariate and multivariate analyses of sexual dimorphism revealed that a greater proportion of between sex variation (sexual dimorphism) was attributable to variation in shape than in size. Significant differences were found in the scores for males and females for the first two principal components. PCI (shape) accounted for 44.25% of the variation and PC2 (size) 13.01% of the variation. Although M. segmentata has been attributed with minimal sexual size dimorphism, females were markedly heavier, possibly a reflection of differential reproductive investment between the sexes, but males had markedly longer legs and broader prosoma. The results are discussed with regard to existing theories of natural and sexual selection, particularly those concerning sexual cannibalism and differential life history traits in males and females. Models that attempt to explain the evolution of sexual size dimorphism in spiders and of the web builders in particular, fail to account for the multivariate nature of dimorphism, especially with respect to shape.     

Shape Variation in the Dermatocranium of the Greater Short-Horned Lizard <Emphasis Type="Italic">Phrynosoma hernandesi</Emphasis> (Reptilia: Squamata: Phrynosomatidae)

Dermatocranial shape and horn morphology display great disparity among the species of Phrynosoma. Ontogenetic change in dermatocranial shape in a series of 79 specimens of the short-horned Phrynosoma hernandesi (54F: 25M) was examined using geometric morphometric techniques. A multivariate ANCOVA of Procrustes residuals with sex as a factor and ln(centroid size) as the covariate indicated sexual shape dimorphism. Separate multivariate regressions of Procrustes residuals on ln(centroid size) for each sex indicated that allometry accounts for ~52–54% of the total sample shape variance. Comparisons of ontogenetic shape change between sexes indicate that sexual shape dimorphism is minimal and of uncertain biological significance. Groupings of multivariate regression coefficients by magnitude and sign suggest that allometric integration of the dermatocranium is not uniform over the dermatocranium. Principal component analysis of the landmark configurations corrected for sex and allometry yields a first principal component which describes shape variance concentrated in the posterolateral and posterior regions of the dermatocranium, and again is indicative of non-uniform shape variation over the dermatocranium. Our findings for P. hernandesi indicate that the adult shape of the dermatocranium may contribute to a passive defence against predation. We hypothesize that the complexity in dermatocranial shape demonstrated here for P. hernandesi indicates parcellation of shape variance, which may contribute to explanations of the pronounced dermatocranial disparity exhibited by the species of Phrynosoma.     

Multivariate variation in feral American mink (Mustela vison) from Southern Norway

Multiple group principal component analysis was used to investigate the morphological relationships between feral American mink from three areas of Southern Norway based on 15 skull dimensions. The first principal axis calculated from the variance-covariance matrix of log-transformed data is usually interpreted as a growth axis. In the present analysis, the scores of the first principal axis of males were found to be uncorrelated with estimated age for specimens younger than seven months old, and was interpreted as a growth-free size axis. However, the second component of males was found to reflect mainly growth in the postorbital area. The first component of females was thought to reflect a mixing of growth-out and growth-in size variation. One of the reasons for separating size and size-free variation in the present work was to test whether growth-dependent size influenced the assessment of geographic variation within each sex. The scores on the components reflecting size variation (growthout and growth-in) were, however, not found to deviate significantly between the areas. Discriminant analyses run on the component scores, both with size variation included and excluded, gave the same result. In mean, the mink from Western Norway deviated significantly from the mink from Trondelag and Eastern Norway, while the mink from the two latter areas did not deviate morphometrically. The deviation found was in shape rather than in size of both sexes.     

Normal Thoracic Radiographic Appearance of the Cynomolgus Monkey (Macaca fascicularis)

Background

The cynomolgus monkey (Macaca fascicularis) has been increasingly used as a non-human primate model in biomedical research. As establishing baseline thoracic radiography for the cynomolgus monkey is essential, we tested the hypothesis that age and sex may affect the thoracic radiography parameters of this species.

Methods

Here, 697 healthy cynomolgus monkeys were segregated by sex and age (three age groups: 25–36 months, 37–48 months, 49–60 months). The lung length (LL), maximal interior thoracic depth (TD), maximal interior thoracic breadth (TBr), cardiac silhouette breadth (CBr), cardiothoracic ratio (CR), right and left costophrenic angles (RCA and LCA), and right hilar height ratio (R-HHR) were assessed by chest film. Statistical analysis was applied to examine the effect of age, sex, and age × sex interactions.

Results

Significant effects by age were shown for LL, TD, TBr, CBr, and CR. Significant effects by sex were found for TD, TBr, CBr, CR, and R-HHR. Significant effects by age × sex were observed for TD, TBr, CBr, and CR. Both TD and TBr increased with age in both sexes, and both were significantly higher in males than in females in the group aged 49–60 months. CBr increased with age and was significantly higher in males than in females across all age groups. CR declined with age and was significantly higher in males than females across all age groups, and CR was similar or slightly higher relative to those previously found in other non-human primate species. As to the other parameters with no significant sex nor age-related differences, the R-HHR was greater than 1.00, and the angulation of bilateral costophrenic angles were sharp.

Conclusions

The thoracic radiographic parameters for the healthy cynomolgus monkey presented here should prove useful in veterinary practice, research involving non-human primate models of respiratory or cardiovascular disorders, and morphological studies on cynomolgus monkeys.     

Sexual dimorphism in the skull of minks Mustela vison, badgers Meles meles and otters Lutra lutra

Multiple-group principal component analysis and discriminant analysis were used to investigate the morphological differences between adult skulls of male and female minks, badgers and otters from Norway. The first principal component axis, calculated from the variance-covariance matrix of log-transformed data, was interpreted as a growth-free size axis in all three species, while the other components were interpreted as representing shape. Having largely separated size and shape variation, these two aspects of sexual dimorphism could be studied. The standardized component scores were subjected to an analysis of variance and discriminant analyses were performed on size-in and size-out data. Sexual dimorphism was disclosed on eight of the 12 components in minks and on seven of the 12 components in badgers and otters. In mink the multivariate differences were more due to size than to shape, whereas in badgers and otters most of the multivariate differences were due to shape, but the differences in size were also significant. The shape dimorphism was shown to be functionally related to jaw and neck muscles. The results were discussed in relation to recent theories to explain the evolutionary significance of sexual dimorphism in body size of mustelids. It was concluded that these theories do not fully explain the dimorphism found in the skulls of the moderately dimorphic badger and otter.     

Growth and allometry of external morphology in Darwin's finches (Geospiza) on Isla Daphne Major, Galapagos

Seven widely used avian external morphological measurements were examined, mainly in Geospiza fortis and G. scandens on Isla Daphne Major, Galapagos. Geospiza grow more slowly than other "finches", and smaller Geospiza species grow relatively more slowly than larger Geospiza . Weight, wing chord and tarsus grow quickly, while bill characters grow more slowly. This pattern of relative growth (dynamic allometry) is reversed in the static allometry of adults, where bill characters, particularly bill depth in G. fortis , display strong, positive allometry with respect to weight, wing and tarsus. Static allometric patterns help explain differences in the size, shape and relative variability of different characters in adults, as well as the results of multivariate analyses such as principal components or canonical variates. Several different multivariate techniques arrange five Geospiza populations in a consistent two-dimensional morphological space, with a major axis of overall "body-size", and a minor axi of "bill-pointedeness". An alternative analysis of dynamic and static allometry is also provided, based on multivariate techniques.     

Multivariate and geometric morphometrics in the analysis of sexual dimorphism variation in Podarcis lizards

Podarcis bocagei and P. carbonelli are two closely related lacertid species, very similar morphologically and ecologically. We investigated sexual dimorphism patterns presented by both species in allopatry and in sympatry. Sexual size and shape dimorphism patterns were analyzed using both multivariate and geometric morphometric techniques. Multivariate morphometrics revealed a marked sexual dimorphism in both species--males being larger with more robust habitus and females presenting a longer trunk. General patterns of sexual size dimorphism are not modified in sympatry, although there is evidence for some morphological change in male head size. The application of geometric morphometrics offered a more detailed image of head shape and revealed that males present a more developed tympanic area than do females, while females have a more rounded head. Differences in the degree of sexual shape dimorphism were detected in sympatry, but no consistent patterns were observed. From the results of the study, and based on previous knowledge on the populations studied, we conclude that the morphological differences observed are probably not caused by exploitative competition between the species, but rather appear attributable to the modification of the relative influence of sexual and natural selection on both sexes.     

A multivariate analysis of temporal change in Arikara craniometrics: A methodological approach

We have examined temporal changes in the cranial architecture of Arikara Amerindians from five archaeological sites in South Dakota which span a time period of approximately 230 years (ca. A.D. 1600–1830). We have utilized a multivariate statistical method based on a principal components analysis of the pooled within-groups correlation matrix rather than the more traditional methods of ascertaining morphological relationships, e.g., discriminant functions, Mahalanobis' D², or Penrose's Size and Shape. Our component structure, based on a regional sample and the mathematically simpler principal components analysis, is very similar to the factor structure obtained by Howells (1973) using a world-wide sample and factor analysis proper. This supports the notion of the “universality” of cranial structure. An axis of temporal variation was introduced into the component space by means of multiple regression. This analysis indicates that a substantial portion of the intergroup variation is temporal in nature and that systematic temporal changes occur along the facial height, transverse frontal flatness, and frontal profile flatness components. Earlier analyses of the same material by more conventional methods either did not detect the temporal trends at all or failed to isolate the specific nature of the temporal changes. The success of the present analysis attests to the value of examining morphological relationships by means of principal components.     

Usefulness of the monkey model to investigate the role soy in postmenopausal women's health

Some of the important health issues for postmenopausal women include cardiovascular disease, osteoporosis, breast cancer, and relief of menopausal symptoms. Ovariectomized cynomolgus monkeys (Macaca fascicularis) have many strengths as models for research in this area including a close phylogenetic relationship to humans, similarities in lipid/lipoprotein metabolism and coronary artery anatomy, similar skeletal anatomical and morphological characteristics, mammary glands with similar pathophysiological characteristics, and a 28-day menstrual cycle with similar hormonal fluctuations. Monkeys (macaques) also experience declining ovarian function and irregular menstrual cycles (natural menopause) when they approach 24 to 29 yr of age. However, because of their very short life span after natural menopause, ovariectomized macaques are used to model postmenopausal women. The cynomolgus monkey model has been useful in defining the potential cardiovascular benefits of soy foods and soy supplements; however, it remains unclear whether the observations are generalizable to all women or only to those who, like cynomolgus monkeys, convert the soy isoflavone daidzein to the metabolite equol. Particularly important has been the use of the cynomolgus monkey model to understand the effects of soy on breast health. There is evidence from a cynomolgus monkey trial to suggest that soy/soy phytoestrogens have no estrogen agonist effects for breast. Finally, soy/soy phytoestrogens do not appear to be an adequate alternative to postmenopausal hormone therapy. Nevertheless, important attributes of soy have been identified, and it may have potential as a complementary component to hormone therapy.