A note on sampling chironomids for RNA-based studies of natural populations that retains critical morphological vouchers

The rapid uptake of transcriptomic approaches in freshwater ecology has seen a wealth of data produced concerning the ways in which organisms interact with their environment on a molecular level. Typically, such studies focus either at the community level and so don’t require species identifications, or on laboratory strains of known species identity or natural populations of large, easily identifiable taxa. For chironomids, impediments still exist for applying these technologies to natural populations because they are small-bodied and often require time-consuming secondary sorting of stream material and morphological voucher preparation to confirm species diagnosis. These procedures limit the ability to maintain RNA quantity and quality in such organisms because RNA degrades rapidly and gene expression can be altered rapidly in organisms; thereby limiting the inclusion of such taxa in transcriptomic studies. Here, we demonstrate that these limitations can be overcome and outline an optimised protocol for collecting, sorting and preserving chironomid larvae that enables retention of both morphological vouchers and RNA for subsequent transcriptomics purposes. By ensuring that sorting and voucher preparation are completed within <4 hours after collection and that samples are kept cold at all times, we successfully retained both RNA and morphological vouchers from all specimens. Although not prescriptive in specific methodology, we anticipate that this paper will assist in promoting transcriptomic investigations of the sublethal impact on chironomid gene expression of changes to aquatic environments.

Learning Nonlinear Visual Processing from Natural Images

The paradigm of computational vision hypothesizes that any visual function -- such as the recognition of your grandparent -- can be replicated by computational processing of the visual input. What are these computations that the brain performs? What should or could they be? Working on the latter question, this dissertation takes the statistical approach, where the suitable computations are attempted to be learned from the natural visual data itself. In particular, we empirically study the computational processing that emerges from the statistical properties of the visual world and the constraints and objectives specified for the learning process. This thesis consists of an introduction and 7 peer-reviewed publications, where the purpose of the introduction is to illustrate the area of study to a reader who is not familiar with computational vision research. In the scope of the introduction, we will briefly overview the primary challenges to visual processing, as well as recall some of the current opinions on visual processing in the early visual systems of animals. Next, we describe the methodology we have used in our research, and discuss the presented results. We have included some additional remarks, speculations and conclusions to this discussion that were not featured in the original publications. We present the following results in the publications of this thesis. First, we empirically demonstrate that luminance and contrast are strongly dependent in natural images, contradicting previous theories suggesting that luminance and contrast were processed separately in natural systems due to their independence in the visual data. Second, we show that simple cell -like receptive fields of the primary visual cortex can be learned in the nonlinear contrast domain by maximization of independence. Further, we provide first-time reports of the emergence of conjunctive (corner-detecting) and subtractive (opponent orientation) processing due to nonlinear projection pursuit with simple objective functions related to sparseness and response energy optimization. Then, we show that attempting to extract independent components of nonlinear histogram statistics of a biologically plausible representation leads to projection directions that appear to differentiate between visual contexts. Such processing might be applicable for priming, \ie the selection and tuning of later visual processing. We continue by showing that a different kind of thresholded low-frequency priming can be learned and used to make object detection faster with little loss in accuracy. Finally, we show that in a computational object detection setting, nonlinearly gain-controlled visual features of medium complexity can be acquired sequentially as images are encountered and discarded. We present two online algorithms to perform this feature selection, and propose the idea that for artificial systems, some processing mechanisms could be selectable from the environment without optimizing the mechanisms themselves. In summary, this thesis explores learning visual processing on several levels. The learning can be understood as interplay of input data, model structures, learning objectives, and estimation algorithms. The presented work adds to the growing body of evidence showing that statistical methods can be used to acquire intuitively meaningful visual processing mechanisms. The work also presents some predictions and ideas regarding biological visual processing.

Impact of the invasive rust Puccinia psidii (myrtle rust) on native Myrtaceae in natural ecosystems in Australia

The invasive rust Puccinia psidii (myrtle rust) was detected in Australia in 2010 and is now established along the east coast from southern New South Wales to far north Queensland. Prior to reaching Australia, severe damage from P. psidii was mainly restricted to exotic eucalypt plantations in South America, guava plantations in Brazil, allspice plantations in Jamaica, and exotic Myrtaceous tree species in the USA; the only previous record of widespread damage in native environments is of endangered Eugenia koolauensis in Hawai’i. Using two rainforest tree species as indicators of the impact of P. psidii, we report for the first time severe damage to endemic Myrtaceae in native forests in Australia, after only 4 years’ exposure to P. psidii. A 3-year disease exclusion trial in a natural stand of Rhodamnia rubescens unequivocally showed that repeated, severe infection leads to gradual crown loss and ultimately tree mortality; trees were killed in less than 4 years. Significant (p < 0.001) correlations were found between both incidence (r = 0.36) and severity (r = 0.38) of P. psidii and subsequent crown loss (crown transparency). This provided supporting evidence to conclude a causal association between P. psidii and crown loss and tree mortality in our field assessments of R. rubescens and Rhodomyrtus psidioides across their native range. Assessments revealed high levels of damage by P. psidii to immature leaves, shoots and tree crowns—averaging 76 % (R. rubescens) and 95 % (R. psidioides) crown transparency—as well as tree mortality. For R. psidioides, we saw exceptionally high levels of tree mortality, with over half the trees surveyed dead and 40 % of stands with greater than 50 % tree mortality, including two stands where all trees were dead. Tree mortality was less prevalent for R. rubescens, with only 12 % of trees surveyed dead and two sites with greater than 50 % mortality. Any alternative causal agents for this tree mortality have been discounted. The ecological implications of this are unclear, but our work clearly illustrates the potential for P. psidii to negatively affect Australia’s biodiversity.

The biology and natural history of Tripogon loliiformis (Poaceae, Chloridoideae), an Australian resurrection grass

Tripogon loliiformis is a desiccation-tolerant grass that occurs throughout mainland Australia. There has been recent interest in this species as a model system for understanding desiccation tolerance in a native grass at the structural, molecular and physiological levels. However, not much is known about the biology and natural history of this species, despite its widespread geographic distribution and remarkable capability of withstanding prolonged drying. We provide an overview of the genus by consolidating information from a wide variety of sources. We report a variety of new and interesting observations on the general biology, ecology and desiccation response of T. loliiformis and conclude by highlighting areas for future research.

The upper respiratory tract is a natural reservoir of haemolytic Mannheimia species associated with ovine mastitis

Lamb suckling has been suggested to be an important way of infecting a ewe's udder with different bacteria, including Mannheimia haemolytica. To test the potential role of lambs in transferring Mannheimia species to the ewe’s udder, the restriction endonuclease cleavage patterns of isolates obtained from nasopharyngeal swabs were compared with those obtained from cases of mastitis. Sterile cotton swabs were used to collect nasopharyngeal samples from 50 ewes and 36 lambs from three flocks. M. haemolytica and Mannheimia glucosida as well as haemolytic Mannheimia ruminalis-like organisms were detected in the upper respiratory tract of lambs and ewes. Comparison of the restriction endonuclease cleavage patterns of the isolates suggested that the M. haemolytica isolates obtained from different milk samples from ewes with mastitis were more clonal than those obtained from the nasal swabs. However, some nasal isolates within both Mannheimia species had restriction endonuclease cleavage patterns identical to those obtained from milk samples from ewes with mastitis, indicating that lambs may have a role in transferring these organisms to the udder. More clonality was observed between the M. glucosida isolates than between M. haemolytica isolates.

Natural Hendra Virus Infection in Flying-Foxes - Tissue Tropism and Risk Factors

Hendra virus (HeV) is a lethal zoonotic agent that emerged in 1994 in Australia. Pteropid bats (flying-foxes) are the natural reservoir. To date, HeV has spilled over from flying-foxes to horses on 51 known occasions, and from infected horses to close-contact humans on seven occasions. We undertook screening of archived bat tissues for HeV by reverse transcription quantitative polymerase chain reaction (RT-qPCR). Tissues were tested from 310 bats including 295 Pteropodiformes and 15 Vespertilioniformes. HeV was detected in 20 individual flying-foxes (6.4%) from various tissues including spleen, kidney, liver, lung, placenta and blood components. Detection was significantly higher in Pteropus Alecto and Pconspicillatus, identifying species as a risk factor for infection. Further, our findings indicate that HeV has a predilection for the spleen, suggesting this organ plays an important role in HeV infection. The lack of detections in the foetal tissues of HeV-positive females suggests that vertical transmission is not a regular mode of transmission in naturally infected flying-foxes, and that placental and foetal tissues are not a major source of infection for horses. A better understanding of HeV tissue tropism will strengthen management of the risk of spillover from flying-foxes to horses and ultimately humans.

Maximum likelihood estimation of natural mortality and quantification of temperature effects on catchability of brown tiger prawn (Penaeus esculentus) in Moreton Bay (Australia) using logbook data

It is common to model the dynamics of fisheries using natural and fishing mortality rates estimated independently using two separate analyses. Fishing mortality is routinely estimated from widely available logbook data, whereas natural mortality estimations have often required more specific, less frequently available, data. However, in the case of the fishery for brown tiger prawn (Penaeus esculentus) in Moreton Bay, both fishing and natural mortality rates have been estimated from logbook data. The present work extended the fishing mortality model to incorporate an eco-physiological response of tiger prawn to temperature, and allowed recruitment timing to vary from year to year. These ecological characteristics of the dynamics of this fishery were ignored in the separate model that estimated natural mortality. Therefore, we propose to estimate both natural and fishing mortality rates within a single model using a consistent set of hypotheses. This approach was applied to Moreton Bay brown tiger prawn data collected between 1990 and 2010. Natural mortality was estimated by maximum likelihood to be equal to 0.032 ± 0.002 week−1, approximately 30% lower than the fixed value used in previous models of this fishery (0.045 week−1).

Estimation of higher natural frequencies of polar orthotropic annular plates

A method based on an assumption that the radial bending moment is zero at a nodal circle is shown to yield accurate estimates of natural frequencies corresponding to higher modes of transversely vibrating polar orthotropic annular plates for various combinations of clamped, simply supported and free edge conditions. This method is found to be convenient for the determination of locations of nodal circles as well. Numerical investigations revealed that for small holes, nodal circles tend to move towards the outer edge with increasing number of nodal diameters. For large holes, it has been shown that the plate behaves like a long rectangular strip.

Natural disasters have unexpected impacts on mental health

Radical circumstances (bushfires and natural disasters) flush out the mental illness in society. Whenever there’s a disaster, there’s a rush on hospital admissions for psychiatric problems. But on the whole, the illness is already there. Emergencies naturally make fodder for delusions and the emergency efforts, for mania. Obviously, there are direct mental health consequences – a small rise in post-traumatic stress disorder inevitably follows disaster. This correlates with the severity of the consequences of the disaster (loss of family, friends, animals and property). And there’s usually a big rethink, with about a third of those affected leaving the area permanently. But, for the most part, this isn’t driven by mental health issues, it results from the very real fears about whether living in a fire (or other disaster) zone is worth it.

Natural convection in rectangular cavities at high Rayleigh number

Natural convection in rectangular two-dimensional cavities with differentially heated side walls is a standard problem in numerical heat transfer. Most of the existing studies has considered the low Ra laminar regime. The general thrust of the present research is to investigate higher Ra flows extending into the unsteady and turbulent regimes where the physics is not fully understood and appropriate models for turbulence are not yet established. In the present study the Boussinesq approximation is being used, but the theoretical background and some preliminary results have been obtained[1] for flows with variable properties.

A computer code for enclosed natural convection in the boundary layer regime

A computer code is developed for the numerical prediction of natural convection in rectangular two-dimensional cavities at high Rayleigh numbers. The governing equations are retained in the primitive variable form. The numerical method is based on finite differences and an ADI scheme. Convective terms may be approximated with either central or hybrid differencing for greater stability. A non-uniform grid distribution is possible for greater efficiency. The pressure is dealt with via a SIMPLE type algorithm and the use of a fast elliptic solver for the solenoidal velocity correction field significantly reduces computing times. Preliminary results indicate that the code is reasonably accurate, robust and fast compared with existing benchmarks and finite difference based codes, particularly at high Rayleigh numbers. Extension to three-dimensional problems and turbulence studies in similar geometries is readily possible and indicated.

The laminar boundary layer regime for natural convection of air in a square cavity

Numerical predictions are obtained for laminar natural convection of air in a square two dimensional cavity at high Rayleigh numbers. Proper resolution of the core reveals weak multi-cellular structure which varies in a complex manner as the effects of convection are increased. The end of the steady laminar regime is numerically estimated to occur at Ra=2.2x10^8.

Laminar and turbulent natural convection in rectangular cavities

A computer program has been developed for the prediction of buoyancy-driven laminar and turbulent flow in rectangular air-filled two-dimensional cavities with differentially heated side walls. Laminar flow predictions for a square cavity and Rayleigh numbers from Ra = 10^3 up to the onset of unsteady flow have been obtained. Accurate solutions for Ra = 5 x 10^6, 10^7, 5 x 10^7 and 10^8 are presented and an estimate for the critical Rayleigh number at which the steady laminar flow becomes unsteady is given for this geometry. Numerical predictions of turbulent flow have been obtained for RaH~0(10^9 -10^11 ) and compared with existing experimental data. A previously developed second moment closure model (Behnia et al. 1987) has been used to model the turbulence. Results indicate that a second moment closure model is capable of predicting the observed flow features.

Laminar and turbulent natural convection in rectangular cavities

"This chapter discusses laminar and turbulent natural convection in rectangular cavities. Natural convection in rectangular two-dimensional cavities has become a standard problem in numerical heat transfer because of its relevance in understanding a number of problems in engineering. Current research identified a number of difficulties with regard to the numerical methods and the turbulence modeling for this class of flows. Obtaining numerical predictions at high Rayleigh numbers proved computationally expensive such that results beyond Ra ∼ 1014 are rarely reported. The chapter discusses a study in which it was found that turbulent computations in square cavities can't be extended beyond Ra ∼ O (1012) despite having developed a code that proved very efficient for the high Ra laminar regime. As the Rayleigh number increased, thin boundary layers began to form next to the vertical walls, and the central region became progressively more stagnant and highly stratified. Results obtained for the high Ra laminar regime were in good agreement with existing studies. Turbulence computations, although of a preliminary nature, indicated that a second moment closure model was capable of predicting the experimentally observed flow features."--Publisher Summary

Natural frequencies of circumferentially truncated sector plates with simply supported straight edges

The classical Rayleigh-Ritz method in conjunction with suitable co-ordinate transformations is found to be effective for accurate estimation of natural frequencies of circumferentially truncated circular sector plates with simply supported straight edges. Numerical results are obtained for all the nine combinations of clamped, simply supported and free boundary conditions at the circular edges and presented in the form of graphs. The analysis confirms an earlier observation that the plate behaves like a long rectangular strip as the width of the plate in the radial direction becomes small.