Ancient DNA suggests dwarf and 'giant' emu are conspecific

Background

The King Island Emu (Dromaius ater) of Australia is one of several extinct emu taxa whose taxonomic relationship to the modern Emu (D. novaehollandiae) is unclear. King Island Emu were mainly distinguished by their much smaller size and a reported darker colour compared to modern Emu.

Methodology and Results

We investigated the evolutionary relationships between the King Island and modern Emu by the recovery of both nuclear and mitochondrial DNA sequences from sub-fossil remains. The complete mitochondrial control (1,094 bp) and cytochrome c oxidase subunit I (COI) region (1,544 bp), as well as a region of the melanocortin 1 receptor gene (57 bp) were sequenced using a multiplex PCR approach. The results show that haplotypes for King Island Emu fall within the diversity of modern Emu.

Conclusions

These data show the close relationship of these emu when compared to other congeneric bird species and indicate that the King Island and modern Emu share a recent common ancestor. King Island emu possibly underwent insular dwarfism as a result of phenotypic plasticity. The close relationship between the King Island and the modern Emu suggests it is most appropriate that the former should be considered a subspecies of the latter. Although both taxa show a close genetic relationship they differ drastically in size. This study also suggests that rates of morphological and neutral molecular evolution are decoupled.     

Pelvic limb musculature in the emu Dromaius novaehollandiae (Aves: Struthioniformes: Dromaiidae): Adaptations to high-speed running

Emus provide an excellent opportunity for studying sustained high-speed running by a bird. Their pelvic limb musculature is described in detail and morphological features characteristic of a cursorial lifestyle are identified. Several anatomical features of the pelvic limb reflect the emus' ability for sustained running at high speeds: (1) emus have a reduced number of toes and associated muscles, (2) emus are unique among birds in having a M. gastrocnemius, the most powerful muscle in the shank, that has four muscle bellies, not the usual three, and (3) contribution to total body mass of the pelvic limb muscles of emus is similar to that of the flight muscles of flying birds, whereas the pelvic limb muscles of flying birds constitute a much smaller proportion of total body mass. Generally, the pelvic limb musculature of emus resembles that of other ratites with the notable exception of M. gastrocnemius. The presence and arrangement of four muscle bellies may increase the effectiveness of M. gastrocnemius and other muscles during cursorial locomotion by moving the limb in a cranio-caudal rather than a latero-medial plane. J. Morphol. 238:23–37, 1998. © 1998 Wiley-Liss, Inc.     

Molecular characterization of goose- and chicken-type lysozymes in emu (Dromaius novaehollandiae): evidence for extremely low lysozyme levels in emu egg white

Lysozyme (LZ), a bacteriolytic enzyme, is found in the egg white of many avian eggs and plays an important role in host defense; however, LZ activity in emu (Dromaius novaehollandiae) egg white is exceptionally undetectable. We cloned and characterized emu goose-type LZ (LZG) and chicken-type LZ (LZC) genes. RT-PCR analysis revealed very low LZG gene expression levels and absence of LZC gene expression in the emu oviduct. Sequencing of full-length LZG and LZC cDNAs indicated that their amino acid sequences show high similarities to ostrich LZG and LZC, respectively, with conserved catalytic residues for enzymatic activities. Whereas recombinant emu LZG prepared using Pichia pastoris exhibited similar enzyme activity as ostrich LZG, recombinant emu LZC exhibited significantly higher lytic activity than chicken LZC. We concluded that emus have functional genes for both LZG and LZC like many other avians, and the LZG gene is expressed in oviduct probably as in other ratite, however, its expression levels in egg white were low to be detected.     

Diversity at the major histocompatibility complex Class II in the platypus, Ornithorhynchus anatinus

The platypus (Ornithorhynchus anatinus) is the sole survivor of a previously widely distributed and diverse lineage of ornithorhynchid monotremes. Its dependence on healthy water systems imposes an inherent sensitivity to habitat degradation and climate change. Here, we compare genetic diversity at the major histocompatibility complex (MHC) Class II-DZB gene and 3 MHC-associated microsatellite markers with diversity at 6 neutral microsatellite markers in 70 platypuses from across their range, including the mainland of Australia and the isolated populations of Tasmania, King Island, and Kangaroo Island. Overall, high DZB diversity was observed in the platypus, with 57 DZB β1 alleles characterized. Significant positive selection was detected within the DZB peptide-binding region, promoting variation in this domain. Low levels of genetic diversity were detected at all markers in the 2 island populations, King Island (endemic) and Kangaroo Island (introduced), with the King Island platypuses monomorphic at the DZB locus. Loss of MHC diversity on King Island is of concern, as the population may have compromised immunological fitness and reduced ability to resist changing environmental conditions.     

Abundance and age of viable resting eggs of the calanoid copepod Boeckella poppei Mrázek in sediments: evidence of egg banks in two Antarctic maritime lakes

The existence of egg banks not only ensures the survival of zooplankton through harsh periods, but could also affect microevolutionary dynamics. Whether zooplankton at high latitudes can build up an egg bank in sediments, as occurs at lower latitudes, is still unknown. The distribution and age of viable resting eggs of the calanoid copepod Boeckella poppei Mrázek in sediments of two small freshwater lakes on King George Island were determined by slicing sediment cores at 1-cm intervals. Most viable resting eggs were found near the sediment surface, with abundance sharply declining to very low values at the depth of 5?cm, although eggs were present as deep as 9?cm in the sediments of Yanouhu Lake. The egg abundances in Xihu Lake and Yanouhu Lake were estimated to be 9.2?×?10⁴ and 7.2?×?10⁴?eggs?m^?2, respectively. ²¹⁰Pb dating indicated a relatively constant sedimentation rate (0.023?cm?year^?1) in Xihu Lake, which was used to estimate the mean age (46.8?year), the maximum age (195.7?year), and the mortality rate (1.64?% year^?1) of resting eggs of B. poppei in Xihu Lake. The accumulation of resting eggs with long-term viability in sediments provided the first evidence for the existence of egg banks in two Antarctic maritime lakes. An egg bank may serve as an overwintering strategy for B. poppei in Antarctica, enhance their ability to cope with random extreme changes, and contribute to their broad distribution.     

The reproductive output of sea lice Caligus rogercresseyi under controlled conditions

Gravid females of Caligus rogercresseyi were collected from Atlantic salmon (Salmo salar) from a farm located at Chiloe Island (42°40′S73°15′W), Chile, to obtain information about the reproductive output of this parasite in vitro. The egg strings removed from the females were incubated under controlled conditions to obtain virgin adult females. One female which had mated only once produced eleven generations of eggs strings in a period of 74 days. The first egg strings of the females obtained in vitro were produced at 389 degree days (°D) after egg incubation, while the next generations of eggs strings were produced with a periodicity between 4 and 6 days dependent on the water temperature. The average length of the egg string was 3.1 mm and the mean number of eggs per string was 31. The values recorded in captivity for the egg string length and the number of eggs per string, were lower than the values recorded in gravid females from the field. One female survived for 79 days and males, maintained separately from the females, survived for 60 days.     

The origin of a new focus of infection with Echinococcus granulosus in tasmania

Hydatid cysts were discovered in cattle on King Island, north of Tasmania, where Echinococcus granulosus was thought to have been eradicated. Using enzyme electrophoresis, isolates from King Island were compared genetically with isolates from Tasmania and the mainland of Australia. The genetic distinctness of the King Island isolates make it unlikely that they originated from a recent introduction from either Tasmania or mainland Australia. Alternative possibilities, that the infection resulted from a recent introduction from another source or from previously undetected persistence of E. granulosus on King Island, could not be distinguished from available data.     

Characterization of new microsatellite markers derived from sequence databases for the emu (Dromaius novaehollandiae)

The emu (Dromaius novaehollandiae), a member of ratite family, is native to Australia and has been introduced to other countries worldwide. In this work, 10 polymorphic microsatellite loci were isolated and characterized for emu from public sequences. Polymorphism was surveyed in 22 individuals from two different populations kept in captivity. Between two and 11 alleles were found per locus, and the observed heterozygosity ranged from 0.05 to 0.85, in accordance with expectations. These markers will be useful as tools for detecting levels of genetic variation, reconstructing pedigrees (for quantitative genetic analysis) and identifying markers associated to fitness traits in emu populations.     

East–west genetic differentiation in Musk Ducks (<Emphasis Type="Italic">Biziura lobata</Emphasis>) of Australia suggests late Pleistocene divergence at the Nullarbor Plain

Musk Ducks (Biziura lobata) are endemic to Australia and occur as two geographically isolated populations separated by the Nullarbor Plain, a vast arid region in southern Australia. We studied genetic variation in Musk Duck populations at coarse (eastern versus western Australia) and fine scales (four sites within eastern Australia). We found significant genetic structure between eastern and western Australia in the mtDNA control region (Φ_ST = 0.747), one nuclear intron (Φ_ST = 0.193) and eight microsatellite loci (F_ST = 0.035). In contrast, there was little genetic structure between Kangaroo Island and adjacent mainland regions within eastern Australia. One small population of Musk Ducks in Victoria (Lake Wendouree) differed from both Kangaroo Island and the remainder of mainland eastern Australia, possibly due to genetic drift exacerbated by inbreeding and small population size. The observed low pairwise distance between the eastern and western mtDNA lineages (0.36%) suggests that they diverged near the end of the Pleistocene, a period characterised by frequent shifts between wet and arid conditions in central Australia. Our genetic results corroborate the display call divergence and Mathews’ (Austral Avian Record 2:83–107, 1914) subspecies classification, and confirm that eastern and western populations of Musk Duck are currently isolated from each other.     

Geographical variation in egg size dimorphism in rockhopper penguins

All crested penguins present a unique reversed hatching asynchrony: the larger second-laid egg (B-egg) hatches before the smaller first-laid egg (A-egg). Although both eggs often hatch, the A-chick generally dies of starvation within days after hatching. However, within rockhopper penguins, the population at the Falkland Islands is unique in that some birds manage to raise both chicks. Although it has been suggested that the egg size dimorphism between A- and B-eggs may explain how long both eggs and chicks survive, this hypothesis has never been explicitly tested. We expect that both eggs are retained longer in the less dimorphic clutches than in the more dimorphic ones. In this paper, we have compiled egg measurements for three rockhopper penguin species (Eudyptes chrysocome, E. filholi and E. moseleyi) in order to compare the intra-clutch egg size dimorphism among these species. Furthermore, we have collected new data to compare egg size dimorphism between two populations of E. chrysocome (Falkland Islands versus Staten Island). A-egg volumes are more variable between species and populations than B-egg volumes. E. chrysocome and especially the population from the Falkland Islands produces the largest A-eggs and the least dimorphic eggs. Nevertheless, as differences in A-egg volumes between species and between the populations of Falkland Islands and Staten Island are stronger and more significant than differences in egg dimorphism, we suggest that A-egg volume, more than egg dimorphism, could be one of the factors influencing the prevalence of twins. A large A-egg and/or reduced egg dimorphism is probably necessary to enable rockhopper penguins to raise two chicks, but other reasons may also be involved which enable them to keep both eggs and chicks.     

Propolis with high flavonoid content collected by honey bees from Acacia paradoxa

Honey bees, Apis mellifera var ligustica, on Kangaroo Island, Australia, were found to collect propolis from the sticky exudate on the stem shoots and seed pods of an Australian endemic plant, Acacia paradoxa. Extracts of the plant stem shoots and seed pods, the propolis carried on the legs of bees and freshly collected propolis in hives contained major flavonoid components consisting of 2′,3′,4′-trimethoxychalcone, 2′-hydroxy-3′,4′-dimethoxychalcone, 2′,4′-dihydroxy-3′-methoxychalcone, 5,7-dihydroxy-2,3-dihydroflavonol 3-acetate (pinobanksin 3-acetate) and 5,7-dihydroxy-6-methoxy-2,3-dihydroflavonol 3-acetate, a substance not previously characterized. HPLC and ¹H NMR analyses of the propolis and plant extracts indicated smaller amounts of other flavonoids. A survey of propolis samples from 47 apiary sites widely distributed on Kangaroo Island showed that 15 samples from 6 sites were largely sourced from A. paradoxa.     

Egg parasitoids (Hymenoptera: Mymaridae) of Amrasca biguttula (Ishida) (Hemiptera: Cicadellidae) on Okinawa Island,a pest of okra in Japan

The fairyfly Anagrus (Anagrus) japonicus Sahad (Hymenoptera: Mymaridae) is identified for the first time as an egg parasitoid of the okra leafhopper Amrasca (Sundapteryx) biguttula (Ishida) (Hemiptera: Cicadellidae) on Okinawa Island, Japan. Amrasca biguttula is a serious pest of okra, Abelmoschus esculentus (L.) Moench (Malvaceae), both in Okinawa and Bonin Islands. Female of A. japonicus is redescribed, and its previously unknown male is described, based on the reared specimens from Okinawa. Prior to this study, host associations of A. japonicus were unknown. Another species of Mymaridae, Arescon enocki (Subba Rao and Kaur), also emerged from eggs of A. biguttula on okra in Okinawa, albeit in much smaller numbers.     

Phylogeographic structure of the dwarf snakehead (Channa gachua) around Gulf of Tonkin: Historical biogeography and pronounced effects of sea‐level changes

Geological events, landscape features, and climate fluctuations have shaped the distribution of genetic diversity and evolutionary history in freshwater fish, but little attention has been paid to that around the Gulf of Tonkin; therefore, we investigated the phylogeographic structure of the dwarf snakehead (Channa gachua) on Hainan Island and mainland China, as well as two populations in Vietnam. We attempted to elucidate the origins of freshwater fish in South Hainan by incorporating genetic data from DNA markers on both the mitochondrial cytochrome b gene (cyt b) and the nuclear recombination‐activating gene 1 (RAG‐1). Mitochondrial phylogenetic analysis identified two major lineages (lineages A and B), which may represent separate species. Divergence data suggested that C. gachua populations diverged between 0.516 and 2.376 myr. The divergence of the two cryptic species is congruent with sea‐level rise, which subsequently isolated Hainan from the mainland. During the Pleistocene glaciations, the entire region of the Gulf of Tonkin and the Qiongzhou Strait became part of the coastal plain of the Asian continent, which might have resulted in the current distribution patterns and dispersal routes of C. gachua populations. The formation of three sublineages in lineage A indicated that the Gulf of Tonkin was a geographical barrier between Hainan Island and mainland China but not between Vietnam and Hainan Island. The results of this study may help to elucidate the origins of freshwater fish in South Hainan and the phylogeographic structure of C. gachua.     

The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island wallaby (Macropus eugenii): development and hatching of the egg and the freeliving stages

Smales L. R. The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): development and hatching of the egg and the free living stages. International Journal for Parasitology7: 449–456. Labiostrongylus eugenii (Trichonematidae) occurs in the stomach of the Kangaroo Island Wallaby. Egg morphology is similar to that of other strongyloids. When incubated at 25°C embryogenesis is completed in about 30 h. An incomplete moult occurs within the egg, and larvae hatch at a sheathed second-stage $\text{43}\text{1}\text{2}\text{–83}\text{1}\text{2}$ h later. Development occurred at all temperatures between 2° and 37°C with an optimum about 25°C and an upper limit near 37°C. The hatching process is very rapid, taking about 2 min. It is signalled by increased larval activity followed by a change in shell permeability. The larva hatches at that pole of the shell which has become plastic.The sheathed second-stage larva measures 659.50 ± 22.54 μm by 27.98 ± 1.22 μm. Its internal structures are concealed by a mass of opaque granules which were demonstrated as neutral lipid by oil red O staining. A second incomplete moult at 3–4 days results in a doubly sheathed infective larva from which the lipid gradually disappears. The mouth never appears patent and the larvae neither feed nor grow but rather decrease in size with age. Optimal temperatures for larvae range between 15°–25°C with 37°C about the upper limit. The significance of this developmental pattern is discussed.     

Ventilatory accommodation of oxygen demand and respiratory water loss in a large bird,the emu (Dromaius novaehollandiae), and a re-examination of ventilatory allometry for birds

Ventilation was studied in the emu, a large flightless bird of mass 40kg, within the range of ambient temperatures from-5 to 45°C. Data for the emu and 21 other species were used to calculate allometric relationships for resting ventilatory parameters in birds (breath frequency=13.5 mass^-0.314; tidal volume=20.7 mass^1.0). At low ambient temperatures the ventilatory system must accommodate the increased metabolic demand for oxygen. In the emu this was achieved by a combination of increased tidal volume and increased oxygen extraction. Data from emus sitting and standing at-5°C, when metabolism is 1.5x and 2.6x basal metabolic rate, respectively, indicate that at least in the emu an increase in oxygen extraction can be stimulated by low temperature independent of oxygen demand. At higher ambient temperatures ventilation was increased to facilitate respiratory water loss. The emu achieved this by increased respiratory frequency. At moderate heat loads (30–35°C) tidal volume fell. This is usually interpreted as a mechanism whereby respiratory water loss can be increased without increasing parabronchial ventilation. At 45°C tidal volume increased; however, past studies have shown that CO₂ washout is minimal under these conditions. The mechanism whereby this is possible is discussed.Abbreviations BMR basal metabolic rate - BTPS body temperature, ambient pressure, saturated - EO ₂ oxygen extraction - EWL evaporative water loss - f _R ventilation frequency - RH relative humidity - RHL respiratory heat loss - SEM standard error of the mean - SNK student-Newman-Keuls multiple range test - STPD standard temperature and pressure, dry - T _a ambient temperatures(s) - T _b body temperature(s) - T _ex expired air temperature(s) - T _rh chamber excurrent air temperature - V _J ventilation - VO₂ oxygen consumption - V _T tidal volume - V/Q air ventilation to blood perfusion ratio     

The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): The parasitic stages

Smales L. R. 1977. The life history of Labiostrongylus eugenii, a nematode parasite of the Kangaroo Island Wallaby (Macropus eugenii): the parasitic stages. International Journal for Parasitology7: 457–461. Labiostrongylus eugenii infective larvae ingested by the Kangaroo Island Wallaby exsheathe on the saccular stomach wall, then invade the mucosa causing one or two chronic irritative hyperpastic nodules, each containing a number of larvae. Six to eight weeks later the larvae leave the stomach wall and moult to fourth-stage. The fourth-stage larva is characterized by eight lips, while the oesophagus and anterior intestine assume the adult form. The posterior ends of late fourth-stage larvae become sexually differentiated. Moulting to adult occurs after 6–8 weeks with the first gravid females being observed 3 months later. The prepatent period appears to last about 6 months. Reasons for failure of experimental infection trials and the epidemiological significance of the life cycle were discussed.     

Small population size and extremely low levels of genetic diversity in island populations of the platypus, Ornithorhynchus anatinus

Genetic diversity generally underpins population resilience and persistence. Reductions in population size and absence of gene flow can lead to reductions in genetic diversity, reproductive fitness, and a limited ability to adapt to environmental change increasing the risk of extinction. Island populations are typically small and isolated, and as a result, inbreeding and reduced genetic diversity elevate their extinction risk. Two island populations of the platypus, Ornithorhynchus anatinus, exist; a naturally occurring population on King Island in Bass Strait and a recently introduced population on Kangaroo Island off the coast of South Australia. Here we assessed the genetic diversity within these two island populations and contrasted these patterns with genetic diversity estimates in areas from which the populations are likely to have been founded. On Kangaroo Island, we also modeled live capture data to determine estimates of population size. Levels of genetic diversity in King Island platypuses are perilously low, with eight of 13 microsatellite loci fixed, likely reflecting their small population size and prolonged isolation. Estimates of heterozygosity detected by microsatellites (H(E)= 0.032) are among the lowest level of genetic diversity recorded by this method in a naturally outbreeding vertebrate population. In contrast, estimates of genetic diversity on Kangaroo Island are somewhat higher. However, estimates of small population size and the limited founders combined with genetic isolation are likely to lead to further losses of genetic diversity through time for the Kangaroo Island platypus population. Implications for the future of these and similarly isolated or genetically depauperate populations are discussed.     

千岛湖社鼠种群遗传现状及与生境面积的关系

以千岛湖地区13个岛屿上的社鼠(Niviventer confucianus)种群为研究对象,利用13个微卫星位点分析了13个社鼠种群的遗传现状,并探讨了种群遗传现状与岛屿面积大小之间的关系。研究结果显示,13个社鼠种群的平均观测等位基因数(Na)与平均有效等位基因数(Ne)分别为7.385,5.952,平均期望杂合度(He)与平均观测杂合度(Ho)分别为0.819,0.930,平均多态信息含量(PIC)为0.760,说明13个社鼠种群均具有较高的遗传多样性。13个种群间的分化系数(Fst)为0.053,表明种群间的遗传分化较小。3个面积较大且相近岛屿上社鼠种群,即高仙阁种群 (Gaoxiange Island,G);乌石T岛种群 (Wushi T Island,WT);东门岛种群 (Dongmen Island,DM)间的Fst值均小于0.05,表明种群间无分化,而小岛种群与大岛种群间的分化却比较明显。此外,一元回归结果显示,多态信息含量(PIC)和Shannon多样性指数(I)与岛屿面积大小之间均存在显著地正相关关系。以上结果说明,在社鼠种群进化过程中,生境面积的缩小可能使种群的遗传结构发生改变,并有可能导致种群的快速进化。