Diving behaviour, dive cycles and aerobic dive limit in the platypus Ornithorhynchus anatinus

We investigated the diving behaviour, the time allocation of the dive cycle and the behavioural aerobic dive limit (ADL) of platypuses (Ornithorhynchus anatinus) living at a sub-alpine Tasmanian lake. Individual platypuses were equipped with combined data logger-transmitter packages measuring dive depth. Mean dive duration was 31.3 s with 72% of all dives lasting between 18 and 40 s. Mean surface duration was 10.1 s. Mean dive depth was 1.28 m with a maximum of 8.77 m. Platypuses performed up to 1600 dives per foraging trip with a mean of 75 dives per hour. ADL was estimated by consideration of post-dive surface intervals vs. dive durations. Only 15% of all dives were found to exceed the estimated ADL of 40 s, indicating mainly aerobic diving in the species. Foraging platypuses followed a model of optimised recovery time, the optimal breathing theory. Total bottom duration or total foraging duration per day is proposed as a useful indicator of foraging efficiency and hence habitat quality in the species.     

Hematologic, plasma biochemical, and other indicators of the health of Tasmanian platypuses (Ornithorhynchus anatinus): predictors of mucormycosis

The fungal disease mucormycosis has affected Tasmanian platypuses for nearly three decades. We investigated the influences of mucormycosis on the hematologic, plasma biochemical, and other indicators of health in free-living platypuses across 18 Tasmanian river catchments. Live trapping enabled sampling of 161 (apparently) healthy and six ulcerated, mucormycosis-affected platypuses in 75 rivers and streams between January 2008 and June 2009. There were no obvious differences in any hematologic or biochemical measures between healthy and mucormycosis-affected platypuses. However, multivariate analysis revealed that ulceration was associated with living at higher altitudes, low tail fat content (high tail fat index), and low trypanosome load. There was evidence of overall lymphocytosis and monocytosis in animals from areas currently affected by mucormycosis, which suggests that some level of immune response to the introduced fungus is now widespread in disease-affected catchments. Animals from currently, historically, and possibly disease-affected catchments had lower neutrophil counts, mean cell volumes, plasma alkaline phosphatase, alanine aminotransferase, and aspartate aminotransferase levels, and higher plasma gamma-glutamyl transpeptidase and platelet counts compared to animals from catchments with no evidence of infection. Reference intervals were generated for all hematologic and biochemical measurements. Since this is the most comprehensive, systematic, and large-scale assessment of the health of the Tasmanian platypus to date, these references intervals should act as the standard against which future studies of platypuses in Tasmania should be compared.     

Structural analysis of viral replicative intermediates isolated from adenovirus type 2-infected HeLa cell nuclei.

Deoxyribonucleoprotein complexes released 17 h postinfection from adenovirus type 1 (Ad2)-infected HeLa cell nuclei were shown by electron microscopy to contain filaments much thicker (about 200 A [20 nm]) than double-stranded DNA (about 20 A [2 nm]). The complexes were partially purified through a linear sucrose gradient, concentrated, and further purified in a metrizamide gradient. The major protein present in the complexes was identified as the 72,000-dalton (72K), adenovirus-coded single-stranded DNA-binding protein (72K DBP). Three types of complexes have been visualized by electron microscopy. Some linear complexes were uniformly thick, and their length corresponded roughly to that of the adenovirus genome. Other linear genome-length complexes appeared to consist of a thick filament connected to a thinner filament with the diameter of double-stranded DNA. Forked complexes consisting of one thick filament connected to a genome-length, thinner double-stranded DNA filament were also visualized. Both thick and thin filaments were sensitive to DNase and not to RNase, but only the thick filaments were digested by the single-strand-specific Neurospora crassa nuclease, indicating that they correspond to a complex of 72K DBP and Ad2 single-stranded DNA. Experiments with anti-72K DBP immunoglobulins indicated that these nucleoprotein complexes, containing the 72K DBP, correspond to replicative intermediates. Both strands of the Ad2 genome were found associated to the 72K DBP. Altogether, our results establish the in vivo association of the 72K DBP with adenovirus single-stranded DNA, as previously suggested from in vitro studies, and support a strand displacement mechanism for Ad2 DNA replication, in which both strands can be displaced. In addition, our results indicate that, late in infection, histones are not bound to adenovirus DNA in the form of a nucleosomal chromatine-like structure.     

Ion etching of human adenovirus 2: structure of the core

The surface of human adenovirus 2 was etched by irradiating intact virions with low-energy (1-keV) Ar+ ions in a Technics Hummer V sputter coater . Viral structures exposed by the etching process were shadowed and then examined in the electron microscope. Periods of etching that were sufficient to reduce the viral diameter by 20 to 30 nm revealed distinct substructural elements in the virion core. Cores were found to consist of a cluster of 12 large, uniformly size spheres which abutted one another in the intact virion. The spheres, for which we suggest the name " adenosomes ," had a diameter of 23.0 +/- 2.3 nm, and they were related to each other by two-, three-, and fivefold axes of rotational symmetry. The results support the view, originally suggested by Brown et al. (J. Virol. 16:366-387, 1975) that the adenovirus 2 core is composed of 12 large spheres packed tightly together in such a way that each is directed toward the vertex of an icosahedron . Such a structure, constructed of 23.0-nm-diameter spheres, would have an outside diameter (vertex-to-vertex distance) of 67.0 nm and a face-to-face distance of 58.2 nm. It could be accommodated inside the icosahedral adenovirus capsid if each large sphere were located beneath a capsid vertex.     

Adenovirus binds to rat brain microtubules in vitro.

We have found by negative staining electron microscopy that when similar concentrations of adenovirus and reovirus (viruses of about the same diameter, 75 to 80 nm, and density, 1.34 to 1.36 g/cm3) were incubated with a carbon support film containing microtubules, 72% of adenovirus on the grid, but only 32% (equivalent to random association) of reovirus, were associated with microtubules. Similar concentrations of both larger and smaller particles, such as polystyrene latex spheres and coliphage f2, also exhibited a low degree of interaction, viz., 17 to 37%, with microtubules. Moreover, 90% of microtubule-associated adenovirus binds to within +/- 4 nm of the edge of microtubules, but lower fractions (again equivalent to a random association) of the other particles bind to the edge of the microtubules. The mechanism behind this phenomenon, which we denote as "edge binding," is presently obscure; however, it provides us with a second, albeit empirical, method to distinguish between the microtubular association of adenovirus and other particles. We found that edge binding of adenovirus also occurred when adenovirus was initially placed on the carbon support film and then incubated with microtubules and when adenovirus and microtubules were mixed prior to placement on the support. In contrast, reovirus or the other particles prepared by similar techniques exhibited a random amount of edge binding. The binding of adenovirus appears to involve the hexon capsomers of the virion since (i) high resolution electron micrographs showed that the edge of the virus was in contact with the edge of the microtubules, and (ii) adenovirions briefly treated with formamide to remove pentons and fibers bind as efficiently as intact virions. Core structures, which were obtained by further formamide degradation of the virion, do not associate with microtubules. These observations support the hypothesis of Dales and Chardonnet (1973) that the transport of adenovirions within infected cells is mediated by interaction with microtubules.     

Energetics of foraging and locomotion in the platypus Ornithorhynchus anatinus

We measured the energy requirements of platypuses foraging, diving and resting in a swim tank using flow-through respirometry. Also, walking metabolic rates were obtained from platypuses walking on a conventional treadmill. Energy requirements while foraging were found to depend on water temperature, body weight and dive duration and averaged 8.48 W kg(-1). Rates for subsurface swimming averaged 6.71 W kg(-1). Minimal cost of transport for subsurface swimming platypuses was 1.85 J N(-1)m(-1) at a speed of 0.4 m s(-1). Aerobic dive limit of the platypus amounted to 59 s. Metabolic rate of platypuses resting on the water surface was minimal with 3.91 W kg(-1) while minimal RMR on land was 2.08 W kg(-1). The metabolic rate for walking was 8.80 W kg(-1) and 10.56 W kg(-1) at speeds of 0.2 m s(-1) and 0.3 m s(-1), respectively. A formula was derived, which allows prediction of power requirements of platypuses in the wild from measurements of body weight, dive duration and water temperature. Platypuses were found to expend energy at only half the rate of semiaquatic eutherians of comparable body sizes during both walking and diving. However, costs of transport at optimal speed were in line with findings for eutherians. These patterns suggest that underwater locomotion of semiaquatic mammals have converged on very similar efficiencies despite differences in phylogeny and locomotor mode.     

A Virus Infection in the Brown Alga Sorocarpus uvaeformis (Lyngbye) Pringsheim (Phaeophyta, Ectocarpales)

Virus-like particles were obsemed in zoospores and less frequentlyin vegetative cells of the filaments of mature plants of thebrown alga Sorocarpus uvaeformls. The particles, measuring approximately170 nm in diameter, are isometric in profile and show threedistinct zones. An electron dense rim (coat), 10 nm in thickness,is separated from a dense core, 110 nm in diameter, by an electronlight space 20 nm in width. When closely packed the particlesare usually separated from each other by a regular halo-likespace. Besides the isometric particles long flexuous structuresof variable length and measuring 75 nm in width were also found.The infection could be induced experimentally in healthy cellsby using either medium prevenient from infected cultures orcrude extracts obtained from infected plants.     

Three-year survey of the epidemiology of rotavirus, enteric adenovirus, and some small spherical viruses including "Osaka-agent" associated with infantile diarrhea

Studies were made by electron microscopy (EM) on the viruses associated with diarrhea of outpatients at a pediatric clinic in Osaka Prefecture during the three year period from 1980 through 1982. The viruses detected by EM by negative staining with phosphotungstic acid (PTA) were classified morphologically into 6 groups: rotavirus, adenovirus and four kinds of small spherical viruses, calicivirus, astrovirus, picornavirus/parvovirus (P/P)-like agent and Osaka-agent. Osaka-agent seems to be a newly identified small virus. It is 35-40 nm in diameter with a fringe of spike-like structures on its surface. Viruses were detected in 181 of the 395 cases of diarrhea (45.8%). Rotavirus was detected in 122 (30.9%) of the total cases and in 67.4% of the virus-positive cases, while other viruses were detected in 15% of the total cases; adenovirus in 23 (6%) and small agents in 36 (9%). Rotavirus infection showed a distinctive seasonal variation, being mainly restricted to cooler months, but infections with other viruses did not show any seasonal variation. The age distribution of patients suggested that infants of 0 to 2 years old are very susceptible to all viruses. Attempts to cultivate these viruses in vitro were successful with only two isolates of adenovirus type 5.     

Biochemical Consequences of Type 2 Adenovirus and Simian Virus 40 Double Infections of African Green Monkey Kidney Cells

African green monkey kidney (AGMK) cells were nonpermissive hosts for type 2 adenovirus although the restriction was not complete; when only 3 plaque-forming units/cell was employed as the inoculum, the viral yield was about 0.1% of the maximum virus produced when simian virus 40 (SV40) enhanced adenovirus multiplication. The viral yield of cells infected only with type 2 adenovirus increased as the multiplicity of infection was increased. Type 2 adenovirus could infect almost all AGMK cells in culture; adenovirus-specific early proteins and DNA were synthesized in most cells, but small amounts of late proteins were made in relatively few cells. Even when cells were infected with both SV40 and adenovirus, only about 50% were permissive for synthesis of adenovirus capsid proteins. Approximately the same quantity of adenovirus deoxyribonucleic acid (DNA) was synthesized in the restricted as in the SV40-enhanced infection. However, in cells infected with SV40 and type 2 adenovirus, replication of SV40 DNA was blocked, multiplication of SV40 was accordingly inhibited, and synthesis of host DNA was not stimulated. To enhance propagation of type 2 adenovirus, synthesis of an early SV40 protein was essential; 50 mug of cycloheximide per ml prevented the SV40-induced enhancement of adenovirus multiplication, whereas 5 x 10(-6)m 5-fluoro-2-deoxyuridine did not abrogate the enhancing phenomenon.     

The morphology and haemodynamics of the rabbit renal artery: evaluation by conventional and contrast-enhanced ultrasonography

The purpose of this study was to test the morphology and haemodynamics of the renal artery in the rabbit as evaluated by conventional and contrast-enhanced ultrasonography (CEUS). The morphology and haemodynamics of the rabbit renal artery, including the diameter, which were measured using B-mode ultrasonography (US), colour Doppler US and CEUS, and systolic velocity, diastolic velocity and resistive index (RI) were measured using pulsed wave Doppler US. CEUS was used to measure the renal artery diameter: 0.21 ± 0.04 cm (right) and 0.21 ± 0.03 cm (left). Values of the main renal artery diameter obtained from CEUS significantly correlated with those of digital subtraction angiography. The blood flow velocity of the right main renal artery was 44.20 ± 8.71/18.92 ± 6.26 cm/s (systolic/diastolic) and 36.30 ± 6.89/17.64 ± 5.58 cm/s (systolic/diastolic), at its origin from the aorta and at the renal hilus, respectively. The blood flow velocity of the left main renal artery was 45.10 ± 8.49/19.00 ± 6.80 cm/s (systolic/diastolic) and 41.70 ± 10.25/19.55 ± 7.90 cm/s (systolic/diastolic), at its origin from the aorta and at the renal hilus, respectively. Conventional US provides a more feasible modality for measuring the morphology and haemodynamics of the rabbit renal artery. CEUS is a more accurate method for measuring diameter. This information on the morphology and haemodynamics of the rabbit renal artery might be helpful for researchers.     

Circadian Activity Rhythms in the Australian Platypus,Ornithorhynchus anatinus (Monotremata)

The Australian platypus, Ornithorhynchus anatinus, is one of three extant genera of the order monotremata. Given the divergent evolutionary lineage of monotremes in relation to more commonly studied animals, it was of interest to determine first, whether platypuses possess endogenous biological pacemakers and, second, general parameters of aquatic activity rhythms under artificial and natural light–dark (LD) cycles. Using a novel recording device, aquatic activity rhythms were measured in three platypuses: a paired male and female studied together, and a single female studied in isolation from other platypuses. Under a constant photic environment, some evidence was found for persistent and free-running rhythmicity, indicating the presence of an endogenous circadian pacemaker in the platypus. Under artificial LD cycles the paired animals exhibited a nocturnal pattern of entrainment, although in the single female considerable variability in entrained phase-relations was found under natural LD cycles. Evidence for a circadian pacemaker in the hypothalamic region of platypuses is also discussed.     

Giant mitochondria in the A cells of the pancreas of a lizard: Varanus niloticus

Pancreatic A cells of the lizard Varanus niloticus are characterized by the presence of two types of mitochondria: (a) normal, small mitochondria (about 0.4 X 1 micron), and (b) giant mitochondria, measuring up to 9 micron in length and 1 micron in diameter. Giant mitochondria show various shapes. Their matrix is filled with tubules, filaments, and dense granules. Transverse sections of tubules are polygonal in shape and about 20 nm in diameter. They are grouped in bundles. The filaments, about 9-10 nm in diameter, are arranged in parallel layers crossing each other at a 57 degree angle. In a closely related species, Varanus exanthematicus, pancreatic A cells do not show these peculiar features.     

用大肠杆菌同源重组获得克隆化重组腺病毒基因组

利用大肠杆菌细胞内质粒间同源重组获得克隆化重组腺病毒基因组 DNA,高效构建携带有外源基因的均一重组腺病毒 .将带有狂犬病毒糖蛋白 (GP)基因和加强型 GFP(enhanced GFP,EGFP)表达盒的重组穿梭质粒 p Ad- Track- CMV/ GP与腺病毒骨架载体质粒 p Ad Easy- 1一起同时电击共转化大肠杆菌 BJ51 83.在 BJ51 83细胞内 ,带有同源序列的重组穿梭质粒与骨架载体可进行同源重组 ,得到以质粒形式存在的克隆化重组腺病毒基因组 p Ad- GP’.以 p Ad- GP’为模板 ,经DNA测序确认 GP基因成功整合入此质粒中的腺病毒基因组 E1区外源基因表达盒中 .线形化的p Ad- GP’转染 2 93细胞后可得到基因组结构均一、在 E1区插入有 GP和 EGFP表达盒的重组腺病毒 ,病毒滴度可达 1× 1 0 8pfu/ ml.电镜下此重组病毒颗粒直径约为 70 nm,略呈球形 ,用荧光显微镜观察感染细胞有很强的 EGFP表达 .实验表明 :利用大肠杆菌同源重组获得克隆化的重组腺病毒基因组 DNA,可高效制备高滴度的均一重组腺病毒     

The fine structure of the parabronchi and the gas exchange area of the Adelie penguin lung

The parabronchi of the Adelie penguin are endowed with wide atria forming pockets between a loose meshwork of bundles of smooth muscle cells lining the parabronchial lumen. The atrial epithelium is of variable thickness and bears numerous microvilli, which are overlain by/or embedded in sheets or whorls of lamellar material ("trilaminar substance", diameter of one lamella 8 ..10 nm) forming layers of very variable thickness. The cells contain either stacks or whorls of this material or roundish lamellated bodies, and are interconnected by desmosomal contacts as well as what presumably represent tight junctions. Underneath the epithelium and within the bundles of muscle cells regularly nerve fibres have been found. The diameter of the morphological air/blood barrier is about 165...210 nm in thin areas, excluding a 12...20 nm thick layer covering the luminal plasma membrane of the air capillary epithelium. The blood capillary endothelium ordinarily is markedly thicker (40...250 nm) than the air capillary epithelium (17...25 nm). The basal lamina between endo- and epithelium is a uniform structure measuring about 95...105 nm. The endothelial cells are interconnected by desmosomal and probably tight junctions.     

Circadian Activity Rhythms in the Australian Platypus, Ornithorhynchus anatinus (Monotremata)

The Australian platypus, Ornithorhynchus anatinus, is one of three extant genera of the order monotremata. Given the divergent evolutionary lineage of monotremes in relation to more commonly studied animals, it was of interest to determine first, whether platypuses possess endogenous biological pacemakers and, second, general parameters of aquatic activity rhythms under artificial and natural light-dark (LD) cycles. Using a novel recording device, aquatic activity rhythms were measured in three platypuses: a paired male and female studied together, and a single female studied in isolation from other platypuses. Under a constant photic environment, some evidence was found for persistent and free-running rhythmicity, indicating the presence of an endogenous circadian pacemaker in the platypus. Under artificial LD cycles the paired animals exhibited a nocturnal pattern of entrainment, although in the single female considerable variability in entrained phase-relations was found under natural LD cycles. Evidence for a circadian pacemaker in the hypothalamic region of platypuses is also discussed.     

Electron Microscopic Observations on the Fine Structure of Cell Walls of Chlamydia psittaci

L-cell cultures were infected with elementary bodies (EB) of meningopneumonitis organisms. Cell walls were prepared from reticulate bodies (RB), which are the intracellular developmental forms into which EB are converted, and from EB at appropriate times after infection. When fragmented EB cell walls were shadowcast with platinum palladium alloy, about one-half of the fragments were seen to be composed of hexagonally arrayed structures on the inner side of the cell wall. When EB cell walls were negatively stained with phosphotungstic acid, they all showed this fine structural array. These macromolecular units were estimated to be about 18 nm in diameter. RB cell walls, harvested at various times after infection, were similarly stained; about 20% of RB walls at 15 hr after infection showed traces of these regular structures, but only 2% of them had the structures at 24 hr. When RB cell walls prepared from penicillin-containing culture were examined, they were observed to be similar to RB without penicillin. When EB cell walls were treated with formamide at 160 C, and then centrifuged in a 10 to 40% potassium tartrate density gradient, hexagonal particles about 20 nm in diameter were obtained as a middle band in the gradient column. These particles were not obtained from RB cell walls harvested from cultures with or without penicillin. It is concluded that the particles are macromolecular subunits located on the inner side of the EB cell walls, that the subunits probably provide the structural rigidity found in the EB, and that their synthesis is inhibited by penicillin.     

Tubular structures occurring in cells infected with herpesviruses

Certain viruses belonging to the Herpesviridae family generate tubular structures of three distinct size ranges late in the growth cycle. The smallest tubules are about 10 to 20 nm in diameter, and are restricted to the cell nucleus, tending to form palisades of lattice-like structures. The medium sized tubular structures are about the width of a core particle, measuring 55 to 65 nm in diameter, while the larger tubular structures are the same diameter as viral capsids, about 80 to 100 nm. Both are rigid capsomered structures, which are sometimes encountered outside the cell nucleus budding onto spherical particles. The available data on some properties of the various tubular structures, as well as speculations concerning their nature and significance are briefly reviewed. Their importance is emphasized for antigen and vaccine production, as well as in differentiating herpesviruses by electron microscopy.     

Topography and functional information of plasma membrane

By using atomic force microscope (AFM), the topography and function of the plasmalemma surface of the isolated protoplasts from winter wheat mesophyll cells were observed, and compared with dead protoplasts induced by dehydrating stress. The observational results revealed that the plasma membrane of living protoplasts was in a state of polarization. Lipid layers of different cells and membrane areas exhibited distinct active states. The surfaces of plasma membranes were unequal, and were characterized of regionalisation. In addition, lattice structures were visualized in some regions of the membrane surface. These typical structures were assumed to be lipid molecular complexes, which were measured to be 15.8±0.09 nm in diameter and 1.9±0.3 nm in height. Both two-dimensional and three-dimensional imaging showed that the plasmalemma surfaces of winter wheat protoplasts were covered with numerous protruding particles. In order to determine the chemical nature of the protruding particles, living protoplasts were treated by proteolytic enzyme. Under the effect of enzyme, large particles became relatively looser, resulting that their width was increased and their height decreased. The results demonstrated that these particles were likely to be of protein nature. These protein particles at plasmalemma surface were different in size and unequal in distribution. The diameter of large protein particles ranged from 200 to 440 nm, with a central micropore, and the apparent height of them was found to vary from 12 to 40 nm. The diameter of mid-sized protein particles was between 40―60 nm, and a range of 1.8―5 nm was given for the apparent height of them. As for small protein particles, obtained values were 12―40 nm for their diameter and 0.7―2.2 nm for height. Some invaginated pits were also observed at the plasma membrane. They were formed by the endocytosis of protoplast. Distribution density of them at plasmalemma was about 16 pits per 15 μm2. According to their size, we classified the invaginated pits into two types―larger pits measuring 139 nm in diameter and 7.2 nm in depth, and smaller pits measuring 96 nm in diameter and 2.3 nm in depth. On dehydration-induced dead pro-toplasts, the degree of polarization of plasma membranes decreased. Lipid molecular layers appeared relatively smooth, and the quantity of integral proteins reduced a lot. Invaginated pits were still de-tectable at the membrane surface, but due to dehydration-induced protoplast contraction, the orifice diameter of pits reduced, and their depth increased. Larger pits averagely measuring 47.4 nm in di-ameter and 31.9 nm in depth, and smaller pits measuring 26.5 nm in diameter and 43 nm in depth at average. The measured thickness of plasma membranes of mesophyll cells from winter wheat examined by AFM was 6.6―9.8 nm, thicker in regions covered with proteins.