Color variation in museum specimens of birds: effects of stress, pigmentation, and duration of storage

Animal coloration often serves as a signal to others that may communicate traits about the individual such as toxicity, status, or quality. Colorful ornaments in many animals are often honest signals of quality assessed by mates, and different colors may beproduced by different biochemical pigments. Investigations of the mechanisms responsible for variation in color expression among birds are best when including a geographically and temporally broad sample. In order to obtain such a sample, studies such as this often use museum specimens; however, in order for museum specimens toserve as an accurate replacement, they must accurately represent living birds, or we must understand the ways in which they differ. In this thesis, I investigated the link between feather corticosterone, a hormone secreted in response to stress, and carotenoid-basedcoloration in the Red-winged Blackbird (Agelaius phoeniceus) in order to explore a mechanistic link between physiological state and color expression. Male Red-winged Blackbirds with lower feather corticosterone had significantly brighter red epaulets than birds with higher feather corticosterone, while I found no significant changes in red chroma. I also performed a methodological comparison of color change in museum specimens among different pigment types (carotenoid and psittacofulvin) and pigments in different locations in the body (feather and bill carotenoids) in order to quantify colorchange over time. Carotenoids and psittacofulvins showed significant reductions in red brightness and chroma over time in the collection, and carotenoid color changed significantly faster than psittacofulvin color. Both bill and feather carotenoids showed significant reductions in red brightness and red chroma over time, but change of both red chroma and red brightness occurred at a similar rate in feathers and bills. In order to use museum specimens of ecological research on bird coloration specimen age must be accounted for before the data can be used; however, once this is accomplished, museum- based color data may be used to draw conclusions about wild populations.

Status of the extinct giant lacertid lizard Gallotia simonyi simonyi (Reptilia: Lacertidae) assessed using mtDNA sequences from museum specimens

[EN] Gallotia simonyi (Steindachner, 1889) is a member of an endemic Canary Island genus and is among the largest of the approximately 250 recent species of lacertid lizards. Although Steindachner gave the locality of the types as the Roques de Salmor, off El Hierro island, previous workers (Urusaustegui, 1983; Manrique & Saavedra, 1873; Viera & Clavijo, 1983) and subsequent ones (Machado, 1985) regard the more western of the two rocks in the group, the Roque Chico de Salmor, as the actual source of material. The Roque Chico is 37 m high with a surface area of less than 10,000 m2 and lies 830 m from the northern coast of El Hierro (Machado, 1985).

Suitability of DNA extracted from archival specimens of fruit-eating bats of the genus Artibeus (Chiroptera, Phyllostomidae) for polymerase chain reaction and sequencing analysis

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Converse Bergmann cline in a eucalyptus herbivore, paropsis atomaria olivier (coleoptera: chrysomelidae) : phenotypic plasticity or local adaptation?

Aim To measure latitude-related body size variation in field-collected Paropsis atomaria Olivier (Coleoptera: Chrysomelidae) individuals and to conduct common-garden experiments to determine whether such variation is due to phenotypic plasticity or local adaptation. Location Four collection sites from the east coast of Australia were selected for our present field collections: Canberra (latitude 35°19' S), Bangalow (latitude 28°43' S), Beerburrum (latitude 26°58' S) and Lowmead (latitude 24°29' S). Museum specimens collected over the past 100 years and covering the same geographical area as the present field collections came from one state, one national and one private collection. Methods Body size (pronotum width) was measured for 118 field-collected beetles and 302 specimens from collections. We then reared larvae from the latitudinal extremes (Canberra and Lowmead) to determine whether the size cline was the result of phenotypic plasticity or evolved differences (= local adaptation) between sites. Results Beetles decreased in size with increasing latitude, representing a converse Bergmann cline. A decrease in developmental temperature produced larger adults for both Lowmead (low latitude) and Canberra (high latitude) individuals, and those from Lowmead were larger than those from Canberra when reared under identical conditions. Main conclusions The converse Bergmann cline in P. atomaria is likely to be the result of local adaptation to season length.

A new dasyurid marsupial from Kroombit Tops, south-east Queensland, Australia : the Silver-headed Antechinus, Antechinus argentus sp. nov. (Marsupialia: Dasyuridae)

Antechinus argentus sp. nov. is currently only known from the plateau at the eastern escarpment of Kroombit Tops National Park, about 400km NNW of Brisbane and 60km SSW of Gladstone, south-east Queensland, Australia. Antechinus flavipes (Waterhouse) is also known from Kroombit Tops NP, 4.5km W of the nearest known population of A. argentus; A. mysticus Baker, Mutton and Van Dyck has yet to be found within Kroombit Tops, but is known from museum specimens taken at Bulburin NP, just 40km ESE, as well as extant populations about 400km to both the south-east and north-west of Kroombit NP. A. argentus can be easily distinguished in the field, having an overall silvery/grey appearance with much paler silver feet and drabber deep greyish-olive rump than A. flavipes, which has distinctive yellow-orange toned feet, rump and tail-base; A. argentus fur is also less coarse than that of A. flavipes. A. argentus has a striking silver-grey head, neck and shoulders, with pale, slightly broken eye-rings, which distinguish it from A. mysticus which has a more subtle greyish-brown head, pale buff dabs of eyeliner and more colourful brownish-yellow rump. Features of the dentary can also be used for identification: A. argentus differs from A. flavipes in having smaller molar teeth, as well as a narrower and smaller skull and from A. mysticus in having on average a narrower snout, smaller skull and dentary lengths and smaller posterior palatal vacuities in the skull. A. argentus is strongly divergent genetically (at mtDNA) from both A. flavipes (9.0–11.2%) and A. mysticus (7.2–7.5%), and forms a very strongly supported clade to the exclusion of all other antechinus species, in both mtDNA and combined (mtDNA and nDNA) phylogenies inferred here. We are yet to make detailed surveys in search of A. argentus from forested areas to the immediate east and north of Kroombit Tops. However, A. mysticus has only been found at these sites in low densities in decades past and not at all in several recent trapping expeditions conducted by the authors. With similar habitat types in close geographic proximity, it is plausible that A. argentus may be found outside Kroombit. Nevertheless, it is striking that from a range of surveys conducted at Kroombit Tops in the last 15 years and intensive surveys by the authors in the last 3 years, totalling more than 5 080 trap nights, just 13 A. argentus have been captured from two sites less than 6 km apart. If this is even close to the true geographic extent of the species, it would possess one of the smallest distributions of an Australian mammal species. With several threats identified, we tentatively recommend that A. argentus be listed as Endangered, pending an exhaustive trapping survey of Kroombit and surrounds.

Concordant molecular and phenotypic data delineate new taxonomy and conservation priorities for the endangered mountain yellow-legged frog

The mountain yellow-legged frog Rana muscosa sensu lato, once abundant in the Sierra Nevada of California and Nevada, and the disjunct Transverse Ranges of southern California, has declined precipitously throughout its range, even though most of its habitat is protected. The species is now extinct in Nevada and reduced to tiny remnants in southern California, where as a distinct population segment, it is classified as Endangered. Introduced predators (trout), air pollution and an infectious disease (chytridiomycosis) threaten remaining populations. A Bayesian analysis of 1901 base pairs of mitochondrial DNA confirms the presence of two deeply divergent clades that come into near contact in the Sierra Nevada. Morphological studies of museum specimens and analysis of acoustic data show that the two major mtDNA clades are readily differentiated phenotypically. Accordingly, we recognize two species, Rana sierrae, in the northern and central Sierra Nevada, and R. muscosa, in the southern Sierra Nevada and southern California. Existing data indicate no range overlap. These results have important implications for the conservation of these two species as they illuminate a profound mismatch between the current delineation of the distinct population segments (southern California vs. Sierra Nevada) and actual species boundaries. For example, our study finds that remnant populations of R. muscosa exist in both the southern Sierra Nevada and the mountains of southern California, which may broaden options for management. In addition, despite the fact that only the southern California populations are listed as Endangered, surveys conducted since 1995 at 225 historic (1899-1994) localities from museum collections show that 93.3% (n=146) of R. sierrae populations and 95.2% (n=79) of R. muscosa populations are extinct. Evidence presented here underscores the need for revision of protected population status to include both species throughout their ranges.

Taxonomic Implications of a Field Study of Morphotypes of Hanuman Langurs (Semnopithecus entellus) in Peninsular India

The Hanuman langur is one of the most widely distributed and morphologically variable non-human primates in South Asia. Even though it has been extensively studied, the taxonomic status of this species remains unresolved due to incongruence between various classification schemes. This incongruence, we believe, is largely due to the use of plastic morphological characters such as coat color in classification. Additionally these classification schemes were largely based on reanalysis of the same set of museum specimens. To bring greater resolution in Hanuman langur taxonomy we undertook a field survey to study variation in external morphological characters among Hanuman langurs. The primary objective of this study is to ascertain the number of morphologically recognizable units (morphotypes) of Hanuman langur in peninsular India and to compare our field observations with published classification schemes. We typed five color-independent characters for multiple adults from various populations in South India. We used the presence-absence matrix of these characters to derive the pair-wise distance between individuals and used this to construct a neighbor-joining (NJ) tree. The resulting NJ tree retrieved six distinct clusters, which we assigned to different morphotypes. These morphotypes can be identified in the field by using a combination of five diagnostic characters. We determined the approximate distributions of these morphotypes by plotting the sampling locations of each morphotype on a map using GIS software. Our field observations are largely concordant with some of the earliest classification schemes, but are incongruent with recent classification schemes. Based on these results we recommend Hill (Ceylon Journal of Science, Colombo 21:277-305, 1939) and Pocock (Primates and carnivora (in part) (pp. 97-163). London: Taylor and Francis, 1939) classification schemes for future studies on Hanuman langurs.

DNA extraction from formalin-fixed Franciscana tissues

The present paper reports the extraction of DNA from formalin-fixed Pontoporia blainvillei tissues. Following the Vachot and Monerot (1996) protocol, fragmented DNA (300-700bp) was extracted from more than 95% of liver and muscle samples. DNA yield in liver samples was significantly higher than in muscle samples (4.574 ± 1.169mg DNA/mg versus 0.808 ± 0.297mg DNA/mg). Similar results were obtained from nine other species of cetaceans and five species of pinnipeds. It is of special interest to have a method that allows the utilisation of museum specimens not originally preserved for genetic studies, which may include rarely available, declining or extinct species. SPANISH: El presente trabajo reporta la extracción de ADN a partir de tejidos formolizados de Pontoporia blainvillei. Siguiendo el protocolo de Vachot y Monerot (1996) se pudo extraer ADN degradado (300-700pb) en más del 95% de las muestras de hígado y músculo analizadas. El rendimiento en ADN fue significativamente mayor en muestras de hígado que en muestras de músculo (4.574 ± 1.169mg DNA/mg tejido húmedo versus 0.808 ± 0.297mg DNA/mg tejido húmedo). Resultados similares se obtuvieron en otras nueve especies de Cetáceos y cinco de Pinnípedos. Resulta de gran interés contar con un método que permita la utilización de especímenes depositados en museos y que no hayan sido originalmente colectados para estudios genéticos, incluyendo especies de difícil obtención, en franca declinación o extintas.

Phylogeny and biogeography of the Petaurista philippensis complex (Rodentia : Sciuridae), inter- and intraspecific relationships inferred from molecular and morphometric analysis

With modified DNA extraction and Purification protocols, the complete cytochrome b gene sequences (1140 bp) were determined from degraded museum specimens. Molecular analysis and morphological examination of cranial characteristics of the giant flying squirrels of Petaurista philippensis complex (P. grandis, P. hainana, and P. yunanensis) and other Petaurista species yielded new insights into long-standing controversies in the Petaurista systematics. Patterns of genetic variations and morphological differences observed in this study indicate that P. hainana, P. albiventer, and P. yunanensis can be recognized as distinct species, and P. grandis and P. petaurista are conspecific populations. Phylogenetic relationships reconstructed by using parsimony, likelihood, and Bayesian methods reveal that, with P. leucogenys as the basal branch, all Petaurista groups formed two distinct clades. Petaurista philippensis, P. hainana, P. yunanensis, and P. albiventer are clustered in the same clade, while P. grandis shows a close relationship to P. petaurista. Deduced divergence times based on Bayesian analysis and the transversional substitution at the third codon suggest that the retreating of glaciers and upheavals or movements of tectonic plates in the Pliocene-Pleistocene were the major factors responsible for the present geographical distributions of Petaurista groups. (c) 2005 Elsevier Inc. All rights reserved.

A new miniature species of the genus Triplophysa (Bahtoridae: Nemacheilinae) from Yunnan, China

During a re-examination of museum specimens of Triplophysa species, some specimens that had been collected from the Jialonghe River in Yunnan Province, China, in April 1975, were identified as a new species. Triplophysa parvus n. sp. can be distinguished

Molecular phylogeny and biogeography of woolly flying squirrel (Rodentia : Sciuridae), inferred from mitochondrial cytochrome b gene sequences

To investigate the genetic diversity between the populations of woolly flying squirrels (Eupetaurus) from the eastern and western extremes of the Himalayas, partial mitochondrial cytochrome b gene sequences (390-810bp) that were determined from the museum specimens were analyzed using maximum parsimony (MP) and maximum likelihood (ML) methods. The molecular data reveal that the two specimens that were collected in northwestern Yunnan (China) are members of the genus Eupetaurus. Reconstructed phylogenetic relationships show that the populations of Eupetaurus in the eastern and western extremes of the Himalayas are two distinct species with significant genetic differences (12%) and diverged about 10.8 million years ago. Eupetaurus is significantly different from Petaurista and Pteromys. The level of estimated pairwise-sequence divergence observed between Eupetaurus and Petaurista or Pteromys is greater than that observed between Eupetaurus and Trogopterus, Belomys, Glaucomys, or Hylopetes. Considering the divergence time of the two Eupetaurus groups, the glaciations and the uplift of the Himalayas and Qinghai-Tibet plateau during the Pliocene-Pleistocene period might be the major factors affecting the present distribution of Eupetaurus along the Himalayas. (C) 2004 Elsevier Inc. All rights reserved.

New data on the morphology and systematic status of Spinitectus petrowi and Spinitectus gigi (Nematoda : Cystidicolidae) parasitic in catfishes in central China

Two little-known nematode species of the genus Spinitectus Fourment, 1883, S. petrowi Belous, 1965 (prevalence 25%, intensity 1-8) and S. gigi Fujita, 1927 (prevalence 10%, intensity 2-3), were collected from the gastrointestinal tract of the yellow catfish, Pelteobagrus fulvidraco (Richardson), from Liangzihu Lake, Hubei Province, central China, in September of 2002. The light and scanning electron microscopical examination of this material, supplemented by a few museum specimens of S. gigi collected from the catfish Clarias fuscus (Lacepede) in southern China, made it possible to study in detail the morphology of these parasite species and to redescribe them. The first species, whose correct name is S. petrowi Belous, 1965, exhibits some morphological features (e.g., unusually short vestibule, shape of pseudolabia and of the left spicule) not found in most other congeners; a unique feature is the presence of peculiar pairs of transversely oriented peg-like cuticular spines with rounded ends on the ventral surface of the female tail. Spinitectus gigi was found to have 28-31 cuticular spines in the first ring, relatively long distances between the 2nd-7th rings of spines, and anterior rings divided into 2 sectors; the excretory pore is located at the level of the 4th ring of cuticular spines; males posses 4 pairs of preanal- and 6 pairs of postanal caudal papillae and a pair of small phasmids. Spinitectus bagri Wang, Wu et Yu, 1993 and S. wulingensis Yu et Wang, 1997 are considered junior synonyms of S. petrowi, whereas S. clariasi Ky, 1971, S. ophicephali Ky, 1971 and S. yuanjiangensis Wang, Wit et Yu, 1997 are regarded to be junior synonyms of S. gigi. Spinitectus petrowi was not previously reported from China.

New records of Paraprionospio (Annelida: Spionidae) from Chinese waters, with the description of a new species

Paraprionospio pinnata (Ehlers, 1901), a poorly known and potentially cosmopolitan polychaete, was examined from museum specimens and from collections in Jiaozhou Bay, the Yellow Sea. New observations indicate that previous Chinese records of P. pinnata are doubtful, and that Chinese waters contain at least three valid species of Paraprionospio, two are known and one is new. Paraprionospio inaequibranchia (Caullery, 1914) and Paraprionospio coora Wilson, 1990, previously misidentified as P. pinnata, are reported from Chinese waters for the first time. Paraprionospio crist'ata. new species, is characterized by having brown pigment patches on the prostomium, ventral crests on chaetigers 9 and 10, dorsal crests on the middle part of the body (from chaetigers 21-23, not beyond chaetiger 29). thin filaments on chaetiger 3, and bifoliate branchial lamellae.

Dental topography and molar wear in Alouatta palliata from Costa Rica.

Paleoprimatologists depend on relationships between form and function of teeth to reconstruct the diets of fossil species. Most of this work has been limited to studies of unworn teeth. A new approach, dental topographic analysis, allows the characterization and comparison of worn primate teeth. Variably worn museum specimens have been used to construct species-specific wear sequences so that measurements can be compared by wear stage among taxa with known differences in diet. This assumes that individuals in a species tend to wear their molar teeth in similar ways, a supposition that has yet to be tested. Here we evaluate this assumption with a longitudinal study of changes in tooth form over time in primates. Fourteen individual mantled howling monkeys (Alouatta palliata) were captured and then recaptured after 2, 4, and 7 years when possible at Hacienda La Pacifica in Costa Rica between 1989-1999. Dental impressions were taken each time, and molar casts were produced and analyzed using dental topographic analysis. Results showed consistent decreases in crown slope and occlusal relief. In contrast, crown angularity, a measure of surface jaggedness, remained fairly constant except with extreme wear. There were no evident differences between specimens collected in different microhabitats. These results suggest that different individual mantled howling monkeys wear their teeth down in similar ways, evidently following a species-specific wear sequence. Dental topographic analysis may therefore be used to compare morphology among similarly worn individuals from different species.

Re-evaluation of Graneledone setebos (Cephalopoda : Octopodidae) and allocation to the genus Megaleledone

The holotype of the Antarctic octopodid Graneledone setebos was re-examined and found to lack the epidermal warts characteristic of the genus Graneledone. It is similar in its large size to another Southern Ocean species, Megaleledone senoi. A comparative study of G. setebos and specimens attributed to M. senoi led us to conclude that M. senoi is a junior synonym of G. setebos. Although M. senoi is not valid, the genus Megaleledone can be separated from other genera by the structure of the radula (which lacks marginal plates) and we therefore consider the genus to be valid. We propose the new combination of Megaleledone setebos and have refigured the beaks and radula of the holotype herein and expanded the description. A search of museum specimens and the literature shows that Megaleledone setebos is more common in Antarctic waters than previously supposed.