Conflict over the molecular clock.

Figure 3 (p. 1310) in the report "A 115-kD polypeptide immunologically related to erythrocyte band 3 is present in Golgi membranes" by S. Kellokumpu et al. (2 Dec., p. 1308) was incorrectly printed. The correct figure is reproduced below.

Convergence and divergence in the evolution of aquatic birds.

Aquatic birds exceed other terrestrial vertebrates in the diversity of their adaptations to aquatic niches. For many species this has created difficulty in understanding their evolutionary origin and, in particular, for the flamingos, hamerkop, shoebill and pelecaniforms. Here, new evidence from nuclear and mitochondrial DNA sequences and DNA-DNA hybridization data indicates extensive morphological convergence and divergence in aquatic birds. Among the unexpected findings is a grouping of flamingos and grebes, species which otherwise show no resemblance. These results suggest that the traditional characters used to unite certain aquatic groups, such as totipalmate feet, foot-propelled diving and long legs, evolved more than once and that organismal change in aquatic birds has proceeded at a faster pace than previously recognized.

Post-albumin, a foetal-specific rat plasma protein: Purification, physicochemical and immunological studies.

1. The foetal-specific post-albumin of rat plasma was purified by electrophoresis on a Pevicon block. 2. The sedimentation coefficient, diffusion coefficient and molecular weight were determined for this protein and found to be similar to those of adult rat plasma albumin. 3. Foetal post-albumin and adult albumin were compared immunologically and shown, with rabbit antisera, to share no antigenic determinants, suggesting different genetic control of the production of each. 4. It is suggested that the disappearance of post-albumin in the newborn animal may result from the disappearance of haemopoietic tissue from the rat liver with advancing age.

Rates of single-copy DNA evolution in phalangeriform marsupials.

DNA/DNA hybridization was used to investigate the relationships of taxa representing the phalangeriform marsupial families Acrobatidae, Burramyidae, Macropodidae, Petauridae, Phalangeridae, and Pseudocheiridae and (as an outgroup) the bandicoot family Peramelidae. In the course of this, a marked rate slowdown was noted in the burramyid lineage represented by Cercartetus caudatus; ANOVA (with Tukey's test) and F-ratio tests of both corrected and uncorrected data matrices confirmed this rate disparity. As burramyids are small, short-generation-time phalangeriforms, these data present a striking counterexample to the common view that rates of change in DNA sequences are inversely correlated with generation time.

Phylogeny through brain traits: trees generated by neural characters.

Phylogenetic trees were computed by the Wagner algorithm from data on up to 15 brain characters scored on 154 specimens of 134 mammalian species. Because the data were not complete on all specimens, only one tree, of 18 taxa, was generated on all 15 features; a tree of 99 species was computed from 10 characters, and trees of 38 species from 10 and 12. The 38-taxon trees were considered best because they preserved most completely the integrity of mammalian orders. All trees consistently separated the subclasses of mammals and suggested that rodents, insectivores, and the tree shrew were most derived on the basis of brain characters. The trees' shapes are sensitive to small alterations in character scorings, largely because of the relatively few characters available and small differences in the number of states among them.

The platypus is not a rodent: DNA hybridization, amniote phylogeny and the palimpsest theory.

We present DNA-hybridization data on 21 amniotes and two anurans showing that discrimination is obtained among most of these at the class and lower levels. Trees generated from these data largely agree with conventional views, for example in not associating birds and mammals. However, the sister relationships found here of the monotremes to marsupials, and of turtles to the alligator, are surprising results which are nonetheless consistent with the results of some other studies. The Marsupionta hypothesis of Gregory is reviewed, as are opinions about the placement of chelonians. Anatomical and reproductive data considered by Gregory do not unequivocally preclude a marsupial-monotreme special relationship, and there is other recent evidence for placing turtles within the Diapsida. We conclude that the evidential meaning of the molecular data is as shown in the trees, but that the topologies may be influenced by a base-compositional bias producing a seemingly slow evolutionary rate in monotremes, or by algorithmic artefacts (in the case of turtles as well).

Base-compositional biases and the bat problem. II. DNA-hybridization trees based on AT- and GC-enriched tracers.

We conducted a series of parallel DNA-hybridization experiments on a small group of bats (species of Pteropus, Rhinolophus, Noctilio and Pteronotus) and outgroups (Lemur, Cynocephalus, Didelphis), using whole-genome labels and tracers made from extracts enriched with AT and two levels of GC content. FITCH (additive phylogenetic trees) topologies were constructed from the four sets of comparisons, indexed as both delta Tmode and delta NPHs (normalized percentage of hybridization). Based on our previous work showing that the shared AT bias of pteropodids and some microchiropterans may affect the rank-ordering of taxa based on either AT- or GC-rich labels, our expectation was that the resulting trees would show differing topologies when generated from tracers made with the variously enriched DNA extracts. Whereas there was some variation among the trees, most of them grouped the bats together, and almost all paired the representative megachiropteran and rhinolophoid microchiropteran as sister-taxa in contrast to the other microchiropterans. As the pteropodid-rhinolophoid relationship is an unexpected and unlikely one, we attribute this association to an AT bias that was not obviated even by our most GC-rich labels, and suggest that such a bias may compromise the truth of some molecular trees. Accordingly, we believe the broader issue of bat monophyly remains unresolved by DNA-hybridization and probably also by gene-sequencing studies.

DNA hybridization evidence for the principal lineages of hummingbirds (Aves:Trochilidae).

The spectacular evolutionary radiation of hummingbirds (Trochilidae) has served as a model system for many biological studies. To begin to provide a historical context for these investigations, we generated a complete matrix of DNA hybridization distances among 26 hummingbirds and an outgroup swift (Chaetura pelagica) to determine the principal hummingbird lineages. FITCH topologies estimated from symmetrized delta TmH-C values and subjected to various validation methods (bootstrapping, weighted jackknifing, branch length significance) indicated a fundamental split between hermit (Eutoxeres aquila, Threnetes ruckeri; Phaethornithinae) and nonhermit (Trochilinae) hummingbirds, and provided strong support for six principal nonhermit clades with the following branching order: (1) a predominantly lowland group comprising caribs (Eulampis holosericeus) and relatives (Androdon aequatorialis and Heliothryx barroti) with violet-ears (Colibri coruscans) and relatives (Doryfera ludovicae); (2) an Andean-associated clade of highly polytypic taxa (Eriocnemis, Heliodoxa, and Coeligena); (3) a second endemic Andean clade (Oreotrochilus chimborazo, Aglaiocercus coelestis, and Lesbia victoriae) paired with thorntails (Popelairia conversii); (4) emeralds and relatives (Chlorostilbon mellisugus, Amazilia tzacatl, Thalurania colombica, Orthorhyncus cristatus and Campylopterus villaviscensio); (5) mountain-gems (Lampornis clemenciae and Eugenes fulgens); and (6) tiny bee-like forms (Archilochus colubris, Myrtis fanny, Acestrura mulsant, and Philodice mitchellii). Corresponding analyses on a matrix of unsymmetrized delta values gave similar support for these relationships except that the branching order of the two Andean clades (2, 3 above) was unresolved. In general, subsidiary relationships were consistent and well supported by both matrices, sometimes revealing surprising associations between forms that differ dramatically in plumage and bill morphology. Our results also reveal some basic aspects of hummingbird ecologic and morphologic evolution. For example, most of the diverse endemic Andean assemblage apparently comprises two genetically divergent clades, whereas the majority of North American hummingbirds belong a single third clade. Genetic distances separating some morphologically distinct genera (Oreotrochilus, Aglaiocercus, Lesbia; Myrtis, Acestrura, Philodice) were no greater than among congeneric (Coeligena) species, indicating that, in hummingbirds, morphological divergence does not necessarily reflect level of genetic divergence.

Brain traits through phylogeny: evolution of neural characters.

We have previously derived a hypothetical tree of the lines of mammalian descent, based upon a comprehensive numerical taxonomic cross-analysis of primitive and derived states of 15 brain traits in 38 representative species. In this communication we use this tree to describe the probable sequence of changes that have taken place in phylogenetic history. 2 characters proved to be multiply convergent, occurring in parallel in several disparate lines of descent. The remaining 9 characters each appeared in ancestors of one or another of the lineages and characterize related progeny.

Phylogeny through brain traits. Relation of lateral olfactory tract fibers to the accessory olfactory formation as a palimpsest of mammalian descent.

In mammals the fibers of the dorsal lateral olfactory tract either pass under the accessory olfactory formation, or they penetrate through it separating the internal granule cells from the output cells. The use of this trait as a phylogenetic indicator in 181 specimens representing 131 species of 16 orders yielded evidence for common ancestry of Insectivora, Chiroptera, Dermoptera, Rodentia, and Primates (including Tupaia), since all share the derived trait, their dorsal lateral olfactory tract fibers passing through the accessory olfactory formation. Carnivora (including Pinnipedia), Hyracoidea, Perissodactyla, and most Artiodactyla share the primitive condition (fibers passing under) with the one order of monotremes and three marsupial orders. The Edentata and Lagomorpha may be separate from the two major placental groups and from each other, or they may represent successive stages in the evolution of the derived state through progressive alterations in the relative chronology of development of olfactory system components, or one or both orders may occupy an ancestral position with respect to the dichotomy within placental mammals.

Phylogeny through brain traits: fifteen characters which adumbrate mammalian genealogy.

Fifteen characters of brain organization are identified for which primitive and derived states can be distinguished, and which are thus useful in reconstructing mammalian phylogeny. The states of the characters are numerically coded in sequence from most primitive to most derived, to facilitate reciprocal comparisons in comprehensive genealogical analyses. The characters include certain features of cerebral circulation, cytoarchitecture, fiber pathways, and sensory projections. Seven characters are shared with nonmammals, a fact which makes then particularly useful for establishing their primitive vs. derived states; the other eight characters are features peculiar to mammals. Ten characters provide impressive reinforcement for traditional major grouping of the species, while five provide bases for new ideas about mammalian relationships.

Phylogeny through brain traits: the distribution of categorizing characters in contemporary mammals.

The varying states of 15 characters of the central neural organization are tabulated as they occur in each of 147 mammalian species. For each character and species, scores are entered designating the primitive or derived state of the character as it is observed in that species. This tabulation provides data in numerically ordered form for multiple analyses of possible phylogenetic relationships which take into account variations in several different characters simultaneously.

Base-compositional biases and the bat problem. III. The questions of microchiropteran monophyly.

Using single-copy DNA hybridization, we carried out a whole genome study of 16 bats (from ten families) and five outgroups (two primates and one each dermopteran, scandentian, and marsupial). Three of the bat species represented as many families of Rhinolophoidea, and these always associated with the two representatives of Pteropodidae. All other microchiropterans, however, formed a monophyletic unit displaying interrelationships largely in accord with current opinion. Thus noctilionoids comprised one clade, while vespertilionids, emballonurids, and molossids comprised three others, successively more closely related in that sequence. The unexpected position of rhinolophoids may be due either to the high AT bias they share with pteropodids, or it may be phylogenetically authentic. Reanalysis of the data with varying combinations of the five outgroups does not indicate a rooting problem, and the inclusion of many bat lineages divided at varying levels similarly discounts long branch attraction as an explanation for the pteropodid-rhinolophoid association. If rhinolophoids are indeed specially related to pteropodids, many synapomorphies of Microchiroptera are called into question, not least the unitary evolution of echolocation (although this feature may simply have been lost in pteropodids). Further, a rhinolophoid-pteropodid relationship--if true--has serious implications for the classification of bats. Finally, among the outgroups, an apparent sister-group relation of Dermoptera and Primates suggests that flying lemurs do not represent the ancestors of some or all bats; yet, insofar as gliding of the type implemented in dermopterans is an appropriate model for the evolution of powered mammalian flying, the position of Cynocephalus in our tree indirectly strengthens the argument that true flight could have evolved more than once among bats.

Phylogeny through brain traits: the mammalian family tree.

Wagner trees based on the analysis of 15 brain characters scored on 154 specimens of 134 mammalian species show consistent patterns of relationship among the taxa, i.e. (1) monotremes are the primitive complement of the group uniting marsupials and placentals; (2) among marsupials, diprotodont Australian forms are more derived; (3) placental mammals divide into two groups of orders, roughly the ferungulates (carnivores, ungulates, and subungulates) and the unguiculates-plus-gliroids (chiropterans, dermopterans, lagomorphs, rodents, primates, and insectivores including elephant shrews); (4) edentates sit at the base of or just before the placental dichotomy; (5) the tree shrew and tarsier show the same pattern of distribution of brain traits as some rodents, as do prosimians and New World monkeys.

DNA-DNA hybridization-based phylogeny for "higher" nonpasserines: reevaluating a key portion of the avian family tree.

A matrix of delta T mode values for 10 birds, including 9 nonpasserines and a suboscine passerine flycatcher, was generated by DNA-DNA hybridization. Within the most derived lineages, all bootstrapped and jackknifed FITCH trees lend strong support to sister-groupings of the two swift families, of hummingbirds to swifts, and of these to a clade containing both owls and night-hawks. The outgroup duck roots the tree between the woodpecker (Piciformes) and the remaining taxa, indicating that Piciformes are among the earliest branches within nonpasserines. However, the succeeding branches to kingfisher, mousebird, and suboscine passerine flycatcher are based on short internodes that are poorly supported by bootstrapping and that give inconsistent results in jackknifing. Although these 3 orders may have arisen through rapid or near-simultaneous divergence, placement of the "advanced" Passeriformes deep within a more "primitive" radiation indicates that nonpasserines are paraphyletic, echoing the same distinction for reptiles with respect to their advanced descendants. Despite significant rate variation among different taxa, these results largely concur with those obtained with the same technique by Sibley and Ahlquist, who used the delta T50H measure and UPGMA analysis. This agreement lends credence to some of their more controversial claims.

Jackknifing of weighted trees: validation of phylogenies reconstructed from distance matrices.

The jackknife strict consensus has been used to detect topological incompatibilities in phylogenetic trees derived from distance data. We here extend this approach to account for branch lengths, as well as topological relationships, when comparing jackknife pseudoreplicates. The average consensus procedure is used to derive a tree reflecting the agreement among the jackknife phylogenies. Combining the average tree with a minimum and a maximum consensus provides information about jackknife tree variability; the range consensus between the minimum and the maximum represents the topological agreement among them. We also demonstrate the effect of the number of pseudoreplicates on the resulting consensus trees by considering single and multiple deletions. The jackknife extensions that we propose are applied to both hypothetical and DNA-DNA hybridization distance data. Consideration of branch lengths reveals greater stability of the jackknife phylogeny than would be demonstrated by a strict consensus based on topology alone.

Pericardial effusion associated with metastatic disease from an unknown primary tumor in a dog.

A 6.5-year-old, spayed female Siberian husky presented with signs of cardiac tamponade and weakness. Pleural, pericardial, and abdominal effusion were identified with radiographs and ultrasound. Pericardiocentesis relieved signs of tamponade, and the dog was clinically improved. Pericardial effusion recurred, and pericardiectomy was performed. Histopathological examination of excised tissues failed to reveal evidence of infectious or neoplastic disease. After pericardiectomy, clinically apparent thoracic effusion persisted. The dog was euthanized, and postmortem histopathological examination revealed emboli of metastatic carcinoma cells in the epicardium. The location of intrathoracic disease in this dog made antemortem diagnosis difficult, if not impossible.

A urethropexy technique for surgical treatment of urethral prolapse in the male dog.

Urethral prolapse is an uncommon condition affecting young male dogs, most commonly English bulldogs. Current described techniques for surgical treatment of urethral prolapse involve manual reduction of prolapsed mucosa and placement of a temporary purse-string suture at the penile tip, or resection of the prolapsed tissue and apposition of urethral and penile mucosa. The incidence of recurrence of urethral prolapse following resection of the prolapse is not known. This report describes a technique for surgical treatment of urethral prolapse in the male dog that minimizes surgical and anesthetic time, is simple to perform, requires minimal equipment, is effective, and is not associated with significant complications or recurrence. Three cases are described.