American elm cultivars: Variation in compartmentalization of infection by Ophiostoma novo‐ulmi and its effects on hydraulic conductivity

Five cultivars and two populations of wild‐type seedlings of American elm (Ulmus americana), 3 and 4 years old, were examined for differences in their abilities to compartmentalize and resist infection by artificially inoculating with Ophiostoma novo‐ulmi. Morphological characteristics of tree defence, often referred to as the compartmentalization of decay in trees model, were used as a conceptual framework, with particular emphasis on the limiting of tangential spread of infection within the xylem and barriers that limit spread outwards to cells formed after infection. To investigate the change in functional xylem over time, 3‐year‐old trees were assessed at multiple time points following inoculation for hydraulic conductivity. Three and four‐year‐old cut trees were placed in 0.1% w/v safranin O for 18 to 24 hr to indicate functional xylem. Transverse sections of the stained stems were used to calculate per cent of sap‐conducting xylem area and the per cent of circumference conducting of first formed cells and later formed cells. At each collection time, trees were assessed for disease severity on a 1–12 scale, based on the percentage of permanent wilt in the crown. There was considerable variation between cultivars in disease severity and their capacity to localize and resist infection. “Prairie Expedition,” which had the lowest disease severity rating in 2015 and the second lowest in 2016, consistently limited the spread of infection into newly formed xylem and had functional xylem around the entire circumference of the stem at 90 days post‐inoculation. “Valley Forge” in 2016 had the lowest overall disease severity rating and was the only cultivar to consistently limit the tangential spread of infection within extant xylem. This research identifies key characteristics that some cultivars have to resist and limit infection and provides new information that can be used in disease screening programmes to evaluate other cultivars and older plant material.     

Epidemic of blindness in kangaroos - evidence of a viral aetiology

OBJECTIVE: To determine the cause of an epidemic of blindness in kangaroos. DESIGN AND PROCEDURES: Laboratory examinations were made of eyes and brains of a large number of kangaroos using serological, virological, histopathological, electron microscopical, immunohistochemical methods, and PCR with cDNA sequencing. In addition, potential insect viral vectors identified during the disease outbreak were examined for specific viral genomic sequences. SAMPLE POPULATION: For histopathological analysis, 55 apparently blind and 18 apparently normal wild kangaroos and wallabies were obtained from New South Wales, Victoria, South Australia, and Western Australia. A total of 437 wild kangaroos and wallabies (including 23 animals with apparent blindness) were examined serologically. RESULTS: Orbiviruses of the Wallal and Warrego serogroups were isolated from kangaroos affected with blindness in a major epidemic in south-eastern Australia in 1994 and 1995 and extending to Western Australia in 1995/96. Histopathological examinations showed severe degeneration and inflammation in the eyes, and mild inflammation in the brains. In affected retinas, Wallal virus antigen was detected by immunohistochemical analysis and orbiviruses were seen in electron microscopy. There was serological variation in the newly isolated Wallal virus from archival Wallal virus that had been isolated in northern Australia. There were also variations of up to 20% in genotype sequence from the reference archival virus. Polymerase chain reactions showed that Wallal virus was present during the epidemic in three species of midges, Culicoides austropalpalis, C dycei and C marksi. Wallal virus nucleic acid was also detected by PCR in a paraffin-embedded retina taken from a blind kangaroo in 1975. CONCLUSION: Wallal virus and perhaps also Warrego virus are the cause of the outbreak of blindness in kangaroos. Other viruses may also be involved, but the evidence in this paper indicates a variant of Wallal virus, an orbivirus transmitted by midges, has the strongest aetiological association, and immunohistochemical analysis implicates it as the most damaging factor in the affected eyes.     

Isolation of an intrinsic precursor to molecular chemisorption

Over the past 70 years, numerous gas-surface adsorption studies have indicated the existence of a weakly bound, mobile intermediate that is a precursor to chemical bond formation. The direct observation and characterization of such a species are presented. Precursor and chemisorbed benzene on a silicon surface were clearly distinguished with the use of a tunable-temperature scanning tunneling microscope. Precursor decay to chemisorption was observed, allowing the salient features of the potential energy surface to be determined.     

Influence of partial or total replacement of glycerol by alternative cryoprotectants in Ghent freezing extender on post‐thaw sperm quality in stallions

Although glycerol is the cryoprotectant most commonly used in stallions, it has also a considerable toxicity for equine sperm. It was the aim of this study to analyse the quality of frozen‐thawed stallion semen after complete or partial replacement of glycerol in the freezing extender by alternative cryoprotectants. We hypothesized that partial or total replacement of glycerol by cryoprotectants occurring in cold‐resistant frog, insect or plant species results in similar or better semen quality after freezing–thawing. As basic medium, the commercial Ghent basic extender was used and either supplemented with glucose and urea, trehalose and proline, or trehalose and betaine. Based on a series of preliminary experiments, semen was frozen in either commercial Ghent cryopreservation extender (Ghent control), Ghent glucose–urea extender or a Ghent combined extender (glucose–urea, trehalose‐betaine and trehalose‐proline; volume ratio of 2:1:2) in a computer‐controlled rate freezer. After freezing–thawing, semen was analysed for motility, membrane integrity, phosphatidylserine translocation, mitochondrial membrane potential and chromatin condensation. No differences between Ghent control and Ghent glucose–urea extender were seen, while all endpoints except DNA integrity were negatively affected in Ghent combined extender (e.g., progressive motility: Ghent 49.2 ± 3.7, Ghent glucose–urea 46.5 ± 4.6, Ghent combined 24.4 ± 2.8%; p < .001). In conclusion, glycerol concentration in a commercial freezing extender for equine spermatozoa can be successfully reduced when urea as an additive cryoprotectant is added and the glucose concentration is elevated. However, total glycerol replacement with urea, betaine, proline and trehalose was less successful.     

Mycosphaerellaceae and Teratosphaeriaceae associated with Eucalyptus leaf diseases and stem cankers in Uruguay

Mycosphaerella leaf diseases represent one of the most important impediments to Eucalyptus plantation forestry. Yet they have been afforded little attention in Uruguay where these trees are an important resource for a growing pulp industry. The objective of this study was to identify species of Mycosphaerellaceae and Teratosphaeriaceae resulting from surveys in all major Eucalyptus growing areas of the country. Species identification was based on morphological characteristics and DNA sequence comparisons for the Internal Transcribed Spacer (ITS) region of the rDNA operon. A total of ten Mycosphaerellaceae and Teratosphaeriaceae were found associated with leaf spots and stem cankers on Eucalyptus. Of these, Mycosphaerella aurantia, M. heimii, M. lateralis, M. scytalidii, Pseudocercospora norchiensis, Teratosphaeria ohnowa and T. pluritubularis are newly recorded in Uruguay. This is also the first report of M. aurantia occurring outside of Australia, and the first record of P. norchiensis and T. pluritubularis in South America. New hosts were identified for Kirramyces gauchensis, M. aurantia, M. marksii, M. lateralis, M. scytalidii, P. norchiensis, T. molleriana, T. ohnowa and T. pluritubularis. Interestingly K. gauchensis, which has been known only as a stem pathogen, was isolated from leaf spots on E. maidenii and E. tereticornis. The large number of Mycosphaerellaceae and Teratosphaeriaceae occurring in Uruguay is disturbing and raises concerns regarding the introduction of new pathogens that could threaten not only Eucalyptus plantations but also native forests.     

Thermally induced phase separation in supported bilayers of glycosphingolipid and phospholipid mixtures

The authors have studied microstructure evolution during thermally induced phase separation in a class of binary supported lipid bilayers using a quantitative application of imaging ellipsometry. The bilayers consist of binary mixtures consisting of a higher melting glycosphingolipid, galactosylceramide (GalCer), which resides primarily in the outer leaflet, and a lower melting, unsaturated phospholipid, 1,2-dilauroyl-sn-glycero-3-phosphocholine (DLPC). Three different bilayer compositions of GalCer/DLPC mixtures at 35:65, 20:80, and 10:90 molar ratios were cooled at controlled rates from their high-temperature homogeneous phase to temperatures corresponding to their phase coexistence regime and imaged in real time using imaging ellipsometry. During the thermotropic course of GalCer gelation, we find that two distinct types of morphological features modulate. First, the formation and growth of chain and fractal-like defects ascribed to the net change in molecular areas during the phase transition. The formation of these defects is consistent with the expected contraction in the molecular area during the liquid crystalline to gel-phase transition. Second, the nucleation and growth of irregularly shaped gel-phase domains, which exhibit either line-tension dominated compact shape or dendritic domains with extended interfaces. Quantifying domain morphology within the fractal framework reveals a close correspondence, and the quantization of the transition width confirms previous estimates of reduced phase transition cooperativity in supported bilayers. A comparison of domain properties indicates that thermal history, bilayer composition, and cooling rate all influence microstructure details including shapes, sizes, and distributions of domains and defects: At lower cooling rates and lower GalCer fractions compact domains form and at higher GalCer fractions (or at higher cooling rates) dendritic domains are evident. This transition of domain morphology from compact shapes to dendritic shapes at higher cooling rates and higher relative fractions of GalCer suggests kinetic control of shape equilibration in these phospho- and glycolipid mixtures.     

Bacterial septic arthritis in 19 dogs

OBJECTIVE: To provide information on the clinical features, diagnosis and treatment of bacterial septic arthritis in dogs. DESIGN: A retrospective study examining case records of all dogs diagnosed with bacterial septic arthritis at Murdoch University Veterinary Hospital between 1988 and 1997. RESULTS: Nineteen dogs were diagnosed with bacterial septic arthritis, which most commonly occurred after surgery involving the stifle joint. Haematogenous infection occurred in only five dogs. Diagnosis was based on clinical signs, joint fluid analysis, radiography, microbiology and/or response to treatment. Chronic lameness was the most common problem at presentation. Analysis of joint fluid invariably revealed large number of nucleated cells, which consisted primarily of neutrophils. In all but one case the neutrophils were nondegenerate. Culture of joint fluid was frequently successful. Staphylococcus spp were the most common bacteria isolated. Treatment involved antimicrobial drugs only in five dogs. Other dogs received antimicrobial drugs in combination with surgical procedures such as joint lavage and removal of nonabsorbable suture material (eight), arthrodesis (two) or amputation (one). Two dogs were euthanased. Most dogs responded well to treatment and were free of signs of septic arthritis at follow-up. CONCLUSION: Bacterial septic arthritis may often be mild and manifest as chronic lameness. Analysis of joint fluid will detect an inflammatory arthropathy but the presence of toxic neutrophils should not be relied on as an indicator of sepsis. Culture of infected joint fluid is likely to be successful if antimicrobials are not given prior to collection and if the sample is inoculated into enrichment broth. Treatment should involve antimicrobial drugs, open-joint lavage and removal of joint prostheses if the infection is associated with previous surgery.     

Nanoengineering of inorganic and hybrid hollow spheres by colloidal templating

Hollow silica and silica-polymer spheres with diameters between 720 and 1000 nanometers were fabricated by consecutively assembling silica nanoparticles and polymer onto colloids and subsequently removing the templated colloid either by calcination or decomposition upon exposure to solvents. Scanning and transmission electron microscopy images demonstrate that the wall thickness of the hollow spheres can be readily controlled by varying the number of nanoparticle-polymer deposition cycles, and the size and shape are determined by the morphology of the templating colloid. The hollow spheres produced are envisioned to have applications in areas ranging from medicine to pharmaceutics to materials science.