William B. Stallcup

“ In analyzing the serological relationships of the species used in this study, it becomes obvious that two or more series of tests must be considered before the birds can be placed in relation to each other. For example, the data presented in Fig. 14 indicate that Spiza and Molothrus show approximately the same degree of serological correspondence to Richmondena. This does not imply necessarily [Pg 193] that Spiza and Molothrus are closely related. If Fig. 15 is examined, it can be determined that Richmondena shows much greater serological correspondence to Spiza than does Molothrus. ”

“ Patterns of leg-musculature were chosen for study because earlier work showed that muscle patterns in the legs of passerine birds are highly stable and vary but little. Variations, therefore, which are consistent in separating groups of species should be significant. Serological techniques were used because the biochemical systems involved seem to be relatively slow to change in response to environmental influences and because the data obtained may be used in a highly objective manner to measure resemblance among species. ”

“ Since the amount of any one antiserum is limited, there is, of necessity, a limit as to the number of birds used in a series of serological tests. Therefore, although the results reveal the actual serological relationships of the individual species, interpretation of the relationships of the taxonomic groups must be undertaken with the realization that such an interpretation is based on tests involving relatively few species of each group. ”

“ The latter muscle is smaller and weaker than in any of the other species and has only one (the posterior) head of origin. The m. flexor hallucis brevis, on the contrary, is larger than in the other birds, compensating, probably, for the small m. flexor hallucis longus. In those differences, however, which separate the carduelines and ploceids from the other birds studied, Vireo resembles, in every instance, the richmondenines, emberizines, tanagers, warblers, and blackbirds. ”

“ Few of the earlier studies were of a quantitative nature, but within the past decade, satisfactory quantitative serological techniques have been developed whereby taxonomic relationships may be estimated. The usefulness of comparative serology in taxonomy has been demonstrated in investigations of many groups wherein results obtained have, in most instances, been compatible with the results obtained by more conventional methods, such as comparative morphology. As Boyden (1942:141) stated, "comparative serology ... is no simple guide to animal relationship." ”

“ For this reason variations involving origin, insertion, or relative position of a muscle, were judged significant. In leg-musculature the Richmondeninae, the Emberizinae, and the Thraupidae resemble one another closely. Several differences in muscle pattern were found, however, which distinguish these groups from the Carduelinae. The leg-musculature of the carduelines closely resembles that of the Ploceidae.
Serological techniques involved the extraction of saline-soluble proteins from the tissues of the species to be studied. These extracts were carefully processed and were used as antigens. ”

“ He suggests, therefore, that the carduelines are not so closely related to the richmondenines and the emberizines as previously has been thought.
Since there are only two cardueline genera, Loximitris and Hesperiphona, endemic to the New World and at least 10 genera with many species endemic to the Old World, Tordoff (1954:15) suggests an Old World origin for the carduelines. He strengthens his argument for this hypothesis by pointing out that in features of the bony palate and in habits the carduelines resemble the estrildines of the Family Ploceidae. ”

“ Most ornithologists are aware that, within the Order Passeriformes, patterns of musculature in the leg have evolved at a slow rate and exhibit little variation within the Order. Differences which do occur, therefore, probably are significant, especially those that are consistent between groups of species. ”

“ Since studies of external morphology have failed in some cases to provide a clear understanding of the relationships of passerine birds, it seems appropriate that attention be given to other morphological features, to physiological features, and to life history studies in an attempt to find other clues to relationships at the family and subfamily levels. ”

“ Saline controls, antiserum controls, and antigen controls were maintained with each test to determine the turbidities inherent in these solutions. These control-turbidities were deducted from the total turbidity developed in each reaction-tube, the resultant turbidity then being considered as that which was caused by the interaction of antigens and antibodies. The turbidities were allowed to develop over a 24-hour period. In the early stages of this investigation the reactions were allowed to take place at 2°C. in order to inhibit bacterial growth. ”

“ The muscles with pinnate fibers seem to be more efficient, each muscle having a greater functional cross section for its bulk than does one with parallel fibers. He assumes that muscles with parallel fibers are more primitive, phylogenetically, than are those with fibers arranged pinnately. ”

“ The rabbit is a variable to be considered in serological tests. Two rabbits exposed to the same antigen, under the same conditions, may produce antisera which differ greatly in their capacities to distinguish different antigens. It is logical to assume, therefore, that two rabbits exposed to different antigens may produce antisera which also differ in this respect. ”

“ Their data indicate, however, that even though changes in the immunological characteristics of proteins are brought about by formolization, the proteins retain enough of their specific chemical characteristics to allow consistent differentiation of species by immunological methods. In the tests which I performed, the relative positions of the precipitin curves, whether native or formolized extracts were involved, remained unchanged (Figs. 10, 11) . All data used in interpretation of the serological relationships were obtained from tests in which formolized antigens of equivalent age were used. ”

“ Most of the birds used were obtained by shooting, but a few were trapped and the exotic species were purchased alive from a pet dealer. When a bird was killed, the entire digestive tract was carefully removed to prevent the escape of digestive enzymes into the tissues and to prevent putrefaction by action of intestinal bacteria. ”

“ Vireo is an example. Furthermore, at least one ploceid (Philetairus) has a small, vestigial tenth primary, whereas some fringillids (Emberizoides, for example) possess a tenth primary which is rather large and ventrally placed (Chapin, 1917:253-254) . ”

“ Fig. 14. Serological relationships of Richmondena, Spiza, and Molothrus. Fig. 15. Serological relationships of Richmondena, Spiza, and Molothrus. Fig. 16. Serological relationships of Carpodacus with the richmondenine-emberizine-thraupid assemblage. Fig. 17. Serological relationships of Carpodacus and Spinus with Richmondena and Junco.
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Figs. ”