Timing of Infection and Development of Alternaria Diseases in the Canopy of Apple Trees

Alternaria leaf blotch and fruit spot of apple caused by Alternaria spp. cause annual losses to the Australian apple industry. Erratic control using protectant fungicides is often experienced and may be due to the lack of understanding of the timing of infection and epidemiology of the diseases. We found that Alternaria leaf blotch infection began about 20 days after bloom (DAB) and the highest disease incidence occurred from 70 to 110 DAB. Alternaria fruit spot infection occurred about 100 DAB in the orchard. Fruit inoculations in planta showed that there was no specific susceptible stage of fruit. Leaves and fruit in the lower canopy of trees showed higher levels of leaf blotch and fruit spot incidence than those in the upper canopy and the incidence of leaf blotch in shoot leaves was higher than in spur leaves. Temperature, relative humidity, and rainfall affected leaf blotch and fruit spot incidence. The gained knowledge on the timing of infection and development of disease may aid in the development of more effective disease management strategies.

The use of plant activators in mango postharvest diseases management

Anthracnose and stem end rots are the main postharvest diseases affecting mangoes in Australia and limiting the shelf life of fruits whenever they are not controlled. The management of these diseases has often relied on the use of fungicide applications either as field spray treatments, postharvest dips or both. Because of concerns with continuous fungicide use, other options for the sustainable management of these diseases are needed. Field trials were conducted to assess the efficacy of three plant activators for the control of these diseases over a 2-year period on 20-year old ‘R2E2’ mango trees in north Queensland. The activators evaluated were: Bion, Kasil and Mangocote. The efficacy of these activators was compared with that of a standard industry field spray program using a combination of fungicides, as well as to un¬treated controls. Conditions favoured good development of the target diseases in both years to be able to differentiate treatment effects. Kasil as a drench was as effective as the standard fungicide program on the management of anthracnose and stem end rots. Bion as foliar sprays showed similar efficacy with its effectiveness comparable with the standard spray program. Both activators had significantly less disease incidences when compared with the untreated control. The third activator, Mangocote was not very effective in controlling the target diseases. Its effect was not significantly better than the untreated controls. The results from this 2-year study suggest that plant activators can play an effective role in mango postharvest disease management. Proper timing could reduce the number of fungicide sprays in an integrated disease management program enabling sustainable yields of quality fruits without the continuous concerns of health and environmental risks from continuous reliance on fungicide use.

Comparison of fungicide dips efficacies for the control of mango postharvest diseases

The shelf-life of mangoes is limited by two main postharvest diseases when not consistently managed. These are anthracnose ( Colletotrichum gloeosporioides) and stem end rots ( Neofusicoccum parvum). The management of these diseases has often relied mainly on the use of fungicide applications either as field spray treatments and/or postharvest dips. Current postharvest dips are under continuous threats because of health concerns and the maximum residue levels allowed on treated fruit continuous to be reviewed and re-assessed. Research needs to keep up with the rate at which changes are occurring following some of these reviews. The recent withdrawal of carbendazin (Spinflo), as a postharvest dip being used to manage stem end rots necessitated the urgent search for a replacement fungicide to manage this disease. A study was therefore undertaken to compare the efficacy of current and potential products that could be used to fill the gap. The following products were evaluated: Carbendazin (Spinflo), Prochloraz (Sportak), Thiobendazole (TBZ) and Fludioxonil (Scholar). These products were tested both under ambient temperatures and as hot dips to identify one that was most effective. Scholar as a hot dip was the most effective product among the ones compared. It effectively controlled both anthracnose and stem end rots at highly significant levels when compared to the untreated control and even Spinflo which is being replaced. As a cold dip, it had some limited effect on anthracnose but had virtually no effect on stem end rots. Based on its performance in these experiments, the product has been recommended for rates and residue studies so that it can be registered as a hot dip for use in controlling postharvest diseases of mangoes.

Networks of stored grain diseases and pests: Strategies for sampling and mitigation

Pathogens and pests of stored grains move through complex dynamic networks linking fields, farms, and bulk storage facilities. Human transport and other forms of dispersal link the components of this network. A network model for pathogen and pest movement through stored grain systems is a first step toward new sampling and mitigation strategies that utilize information about the network structure. An understanding of network structure can be applied to identifying the key network components for pathogen or pest movement through the system. For example, it may be useful to identify a network node, such as a local grain storage facility, through which grain from a large number of fields will be accumulated and move through the network. This node may be particularly important for sampling and mitigation. In some cases more detailed information about network structure can identify key nodes that link two large sections of the network, such that management at the key nodes will greatly reduce the risk of spread between the two sections. In addition to the spread of particular species of pathogens and pests, we also evaluate the spread of problematic subpopulations, such as subpopulations with pesticide resistance. We present an analysis of stored grain pathogen and pest networks for Australia and the United States.

Foliar, fruit and soilborne diseases.

This chapter provides an in-depth review of important diseases affecting avocado production throughout the world. The importance of understanding the interaction of plant pathogens with their avocado host in order for the development of disease management options is also discussed.

Pre-emptive breeding to combine superior eating quality in tropical super sweet corn with resistance to major diseases

This report presents the process and outcomes of a five year project, which employed genetics and breeding approach for integrating disease resistance,agronomy and quality traits that enhances sustainable productivity improvement in sweet corn production. The report outlines a molecular markers based approach to introgress quantitative traits loci that are believed to contribute to resistance to downy mildew, a potentially devastating disease that threatens sweet corn and other similar crops. It also details the approach followed to integrate resistances for other major diseases such as southern rust (caused by Puccinia polysora Underw), Northern Corn Leaf Blight (Exserohilum turcicum) with improved agronomy and eating quality. The report explains the importance of heterosis (hybrid vigour) and combining ability in the development of useful sweet corn hybrids. It also explains the relevance of parental performance to predict its breeding value and the performance of its hybrids.

Novel vaccine acceptance in emerging infectious diseases: a case study of Hendra virus vaccine uptake in Australia

Background: The development of a horse vaccine against Hendra virus has been hailed as a good example of a One Health approach to the control of human disease. Although there is little doubt that this is true, it is clear from the underwhelming uptake of the vaccine by horse owners to date (approximately 10%) that realisation of a One Health approach requires more than just a scientific solution. As emerging infectious diseases may often be linked to the development and implementation of novel vaccines this presentation will discuss factors influencing their uptake; using Hendra virus in Australia as a case study. Methods: This presentation will draw on data collected from the Horse owners and Hendra virus: A Longitudinal cohort study To Evaluate Risk (HHALTER) study. The HHALTER study is a mixed methods research study comprising a two-year survey-based longitudinal cohort study and qualitative interview study with horse owners in Australia. The HHALTER study has investigated and tracked changes in a broad range of issues around early uptake of vaccination, horse owner uptake of other recommended disease risk mitigation strategies, and attitudes to government policy and disease response. Interviews provide further insights into attitudes towards risk and decision-making in relation to vaccine uptake. A combination of quantitative and qualitative data analysis will be reported. Results: Data collected from more than 1100 horse owners shortly after vaccine introduction indicated that vaccine uptake and intention to vaccinate was associated with a number of risk perception factors and financial cost factors. In addition, concerns about side effects and veterinarians refusing to treat unvaccinated horses were linked to uptake. Across the study period vaccine uptake in the study cohort increased to more than 50%, however, concerns around side effects, equine performance and breeding impacts, delays to full vaccine approvals, and attempts to mandate vaccination by horse associations and event organisers have all impacted acceptance. Conclusion: Despite being provided with a safe and effective vaccine for Hendra virus that can protect horses and break the transmission cycle of the virus to humans, Australian horse owners have been reluctant to commit to it. General issues pertinent to novel vaccines, combined with challenges in the implementation of the vaccine have led to issues of mistrust and misconception with some horse owners. Moreover, factors such as cost, booster dose schedules, complexities around perceived risk, and ulterior motives attributed to veterinarians have only served to polarise attitudes to vaccine acceptance.

Priority list of endemic diseases for the red meat industries

This report provides a systematic review of the most economically damaging endemic diseases and conditions for the Australian red meat industry (cattle, sheep and goats). A number of diseases for cattle, sheep and goats have been identified and were prioritised according to their prevalence, distribution, risk factors and mitigation. The economic cost of each disease as a result of production losses, preventive costs and treatment costs is estimated at the herd and flock level, then extrapolated to a national basis using herd/flock demographics from the 2010-11 Agricultural Census by the Australian Bureau of Statistics. Information shortfalls and recommendations for further research are also specified. A total of 17 cattle, 23 sheep and nine goat diseases were prioritised based on feedback received from producer, government and industry surveys, followed by discussions between the consultants and MLA. Assumptions of disease distribution, in-herd/flock prevalence, impacts on mortality/production and costs for prevention and treatment were obtained from the literature where available. Where these data were not available, the consultants used their own expertise to estimate the relevant measures for each disease. Levels of confidence in the assumptions for each disease were estimated, and gaps in knowledge identified. The assumptions were analysed using a specialised Excel model that estimated the per animal, herd/flock and national costs of each important disease. The report was peer reviewed and workshopped by the consultants and experts selected by MLA before being finalised. Consequently, this report is an important resource that will guide and prioritise future research, development and extension activities by a variety of stakeholders in the red meat industry. This report completes Phase I and Phase II of an overall four-Phase project initiative by MLA, with identified data gaps in this report potentially being addressed within the later phases. Modelling the economic costs using a consistent approach for each disease ensures that the derived estimates are transparent and can be refined if improved data on prevalence becomes available. This means that the report will be an enduring resource for developing policies and strategies for the management of endemic diseases within the Australian red meat industry.

Risk assessment for airborne infectious diseases between natural ventilation and split-system air conditioner in a university classroom

Indoor air quality is a critical factor in the classroom due to high people concentration in a unique space. Indoor air pollutant might increase the chance of both long and short-term health problems among students and staff, reduce the productivity of teachers and degrade the student’s learning environment and comfort. Adequate air distribution strategies may reduce risk of infection in classroom. So, the purpose of air distribution systems in a classroom is not only to maximize conditions for thermal comfort, but also to remove indoor contaminants. Natural ventilation has the potential to play a significant role in achieving improvements in IAQ. The present study compares the risk of airborne infection between Natural Ventilation (opening windows and doors) and a Split-System Air Conditioner in a university classroom. The Wells-Riley model was used to predict the risk of indoor airborne transmission of infectious diseases such as influenza, measles and tuberculosis. For each case, the air exchange rate was measured using a CO2 tracer gas technique. It was found that opening windows and doors provided an air exchange rate of 2.3 air changes/hour (ACH), while with the Split System it was 0.6 ACH. The risk of airborne infection ranged between 4.24 to 30.86 % when using the Natural Ventilation and between 8.99 to 43.19% when using the Split System. The difference of airborne infection risk between the Split System and the Natural Ventilation ranged from 47 to 56%. Opening windows and doors maximize Natural Ventilation so that the risk of airborne contagion is much lower than with Split System.

Innate immunity and its control in the pathogenesis of inflammatory rheumatic diseases

The pathogenesis of inflammatory rheumatic diseases, including rheumatoid arthritis (RA) and spondyloarthropathies (SpAs) such as reactive arthritis (ReA), is incompletely understood. ReA is a sterile joint inflammation, which may follow a distal infection caused by Gram-negative bacteria that have lipopolysaccharide (LPS) in their outer membrane. The functions of innate immunity that may affect the pathogenesis, prognosis and treatment of these diseases were studied in this thesis. When compared with healthy controls, whole blood monocytes of healthy subjects with previous ReA showed enhanced capacity to produce TNF, an essential proinflammatory cytokine, in response to adherent conditions (mimicking vascular endothelium made adherent by inflammatory signals) and non-specific protein kinase C stimulation. Also, blood neutrophils of these subjects showed high levels of CD11b, an important adhesion molecule, in response to adherence or LPS. Thus, high responsiveness of monocytes and neutrophils when encountering inflammatory stimuli may play a role in the pathogenesis of ReA. The results also suggested that the known risk allele for SpAs, HLA-B27, may be an additive contributor to the observed differences. The promoter polymorphisms TNF 308A and CD14 (gene for an LPS receptor component) 159T were found not to increase the risk of acute arthritis. However, all female patients who developed chronic SpA had 159T and none of them had 308A, possibly reflecting an interplay between hormonal and inflammatory signals in the development of chronic SpA. Among subjects with early RA, those having the polymorphic TLR4 +896G allele (causing the Asp299Gly change in TLR4, another component of LPS receptor) required a combination of disease-modifying antirheumatic drugs to achieve remission. It is known that rapid treatment response is essential in order to maintain the patients work ability. Hence, +896G might be a candidate marker for identifying the patients who need combination treatment. The production of vascular endothelial growth factor (VEGF), which strongly promotes vascular permeability and angiogenesis that takes place e.g. early in rheumatic joints, was induced by LPS and inhibited by interferon (IFN)-alpha in peripheral blood mononuclear cells. These long-living cells might provide a source of VEGF when stimulated by LPS and migrating to inflamed joints, and the effect of IFN-alpha may contribute to the clinical efficacy of this cytokine in inhibiting joint inflammation.

Nephrin-associated molecules in human glomerular diseases

The glomerular epithelial cells and their intercellular junctions, termed slit diaphragms, are essential components of the filtration barrier in the kidney glomerulus. Nephrin is a transmembrane adhesion protein of the slit diaphragm and a signalling molecule regulating podocyte physiology. In congenital nephrotic syndrome of the Finnish type, mutation of nephrin leads to disruption of the permeability barrier and leakage of plasma proteins into the urine. This doctoral thesis hypothesises that novel nephrin-associated molecules are involved in the function of the filtration barrier in health and disease. Bioinformatics tools were utilized to identify novel nephrin-like molecules in genomic databases, and their distribution in the kidney and other tissues was investigated. Filtrin, a novel nephrin homologue, is expressed in the glomerular podocytes and, according to immunoelectron microscopy, localizes at the slit diaphragm. Interestingly, the nephrin and filtrin genes, NPHS1 and KIRREL2, locate in a head-to-head orientation on chromosome 19q13.12. Another nephrin-like molecule, Nphs1as was cloned in mouse, however, no expression was detected in the kidney but instead in the brain and lymphoid tissue. Notably, Nphs1as is transcribed from the nephrin locus in an antisense orientation. The glomerular mRNA and protein levels of filtrin were measured in kidney biopsies of patients with proteinuric diseases, and marked reduction of filtrin mRNA levels was detected in the proteinuric samples as compared to controls. In addition, altered distribution of filtrin in injured glomeruli was observed, with the most prominent decrease of the expression in focal segmental glomerulosclerosis. The role of the slit diaphragm-associated genes for the development of diabetic nephropathy was investigated by analysing single nucleotide polymorphisms. The genes encoding filtrin, densin-180, NEPH1, podocin, and alpha-actinin-4 were analysed, and polymorphisms at the alpha-actinin-4 gene were associated with diabetic nephropathy in a gender-dependent manner. Filtrin is a novel podocyte-expressed protein with localization at the slit diaphragm, and the downregulation of filtrin seems to be characteristic for human proteinuric diseases. In the context of the crucial role of nephrin for the glomerular filter, filtrin appears to be a potential candidate molecule for proteinuria. Although not expressed in the kidney, the nephrin antisense Nphs1as may regulate the expression of nephrin in extrarenal tissues. The genetic association analysis suggested that the alpha-actinin-4 gene, encoding an actin-filament cross-linking protein of the podocytes, may contribute to susceptibility for diabetic nephropathy.

Childhood-onset mitochondrial diseases : with emphasis on molecular diagnostics of neurological disorders

Childhood-onset mitochondrial diseases comprise a heterogeneous group of disorders, which may manifest with almost any symptom and affect any tissue or organ. Due to challenging diagnostics, most children still lack a specific aetiological diagnosis. The aim of this thesis was to find molecular causes for childhood-onset mitochondrial disorders in Finland. We identified the underlying cause for 25 children, and found three new diseases, which had not been diagnosed in Finland before. These diseases caused severe progressive infantile-onset encephalomyopathies, and were due to defects in mitochondrial DNA (mtDNA) maintenance. Furthermore, the thesis provides the molecular background of Finnish patients with ‘leukoencephalopathy with brain stem and spinal cord involvement and elevated brain lactate’ (LBSL). A new phenotype was identified to be due to mutations in Twinkle, resembling ‘infantile onset spinocerebellar ataxia’ (IOSCA). These mutations caused mtDNA depletion in the liver, thus confirming the essential role of Twinkle in mtDNA maintenance, and expanding the molecular background of mtDNA depletion syndromes. The major aetiology for infantile mitochondrial myopathy in Finland was discovered to be due to mutations in thymidine kinase 2 (TK2). A novel mutation with Finnish ancestry was identified, and a genotype-phenotype correlation with mutation-specific distribution of tissue involvement was found, thus proving that deficient TK2 may cause multi-tissue depletion and impair neuronal function. This work established the molecular diagnosis and advanced the knowledge of phenotypes among paediatric patients with polymerase gamma (POLG) mutations. The patients showed severe early-onset encephalopathy with intractable epilepsy. POLG mutations are not a prevalent cause of children’s ataxias, although ataxia is a major presenting symptom among adults. Our findings indicate that POLG mutations should be investigated even if typical MRI, histochemical or biochemical abnormalities are lacking. LBSL patients showed considerable variation in phenotype despite identical mutations. A common, most likely European, ancestry, and a relative high carrier frequency of these mutations in Finland were discovered; suggesting that LBSL may be a quite common leukoencephalopathy in other populations as well. The results suggest that MRI findings are so unique that the diagnosis of LBSL is possible to make without genetic studies. This thesis work has resulted in identification of new mitochondrial disorders in Finland, enhancing the understanding of the clinical variability and the importance of tissue-specificity of these disorders. In addition to providing specific diagnosis to the patients, these findings give light to the underlying pathogenetic mechanisms of childhood-onset mitochondrial disorders.

The Use of Mesenchymal Stromal Cells in the Treatment of Diseases of the Cornea

As a key component of the ocular surface required for vision, the cornea has been extensively studied as a site for cell and tissue-based therapies. Historically, these treatments have consisted of donor corneal tissue transplants, but cultivated epithelial autografts have become established over the last 15 years as a routine treatment for ocular surface disease. Ultimately, these treatments are performed with the intention of restoring corneal transparency and a smooth ocular surface. The degree of success, however, is often dependent upon the inherent level of corneal inflammation at time of treatment. In this regard, the anti-inflammatory and immuno-modulatory properties of mesenchymal stromal cells (MSC) have drawn attention to these cells as potential therapeutic agents for corneal repair. The origins for MSC-based therapies are founded in part on observations of the recruitment of endogenous bone marrow-derived cells to injured corneas, however, an increasing quantity of data is emerging for MSC administered following their isolation and ex vivo expansion from a variety of tissues including bone marrow, adipose tissue, umbilical cord and dental pulp. In brief, evidence has emerged of cultured MSC, or their secreted products, having a positive impact on corneal wound healing and retention of corneal allografts in animal models. Optimal dosage, route of administration and timing of treatment, however, all remain active areas of investigation. Intriguingly, amidst these studies, have emerged reports of MSC transdifferentiation into corneal cells. Clearest evidence has been obtained with respect to expression of markers associated with the phenotype of corneal stromal cells. In contrast, the evidence for MSC conversion to corneal epithelial cell types remains inconclusive. In any case, the conversion of MSC into corneal cells seems unlikely to be an essential requirement for their clinical use. This field of research has recently become more complicated by reports of MSC-like properties for cultures established from the peripheral corneal stroma (limbal stroma). The relationship and relative value of corneal-MSC compared to traditional sources of MSC such as bone marrow are at present unclear. This chapter is divided into four main parts. After providing a concise overview of corneal structure and function, we will highlight the types of corneal diseases that are likely to benefit from the anti-inflammatory and immuno-modulatory properties of MSC. We will subsequently summarize the evidence supporting the case for MSC-based therapies in the treatment of corneal diseases. In the third section we will review the literature concerning the keratogenic potential of MSC. Finally, we will review the more recent literature indicating the presence of MSC-like cells derived from corneal tissue.

Health-Related Quality of Life, Costs and Treatment of Inflammatory Rheumatic Diseases in Routine Practice : With special emphasis on biological drugs

Rheumatoid arthritis (RA) and other chronic inflammatory joint diseases already begin to affect patients health-related quality of life (HRQoL) in the earliest phases of these diseases. In treatment of inflammatory joint diseases, the last two decades have seen new strategies and treatment options introduced. Treatment is started at an earlier phase; combinations of disease-modifying anti-rheumatic drugs (DMARDs) and corticosteroids are used; and in refractory cases new drugs such as tumour necrosis factor (TNF) inhibitors or other biologicals can be started. In patients with new referrals to the Department of Rheumatology of the Helsinki University Central Hospital, we evaluated the 15D and the Stanford Health Assessment Questionnaire (HAQ) results at baseline and approximately 8 months after their first visit. Altogether the analysis included 295 patients with various rheumatic diseases. The mean baseline 15D score (0.822, SD 0.114) was significantly lower than for the age-matched general population (0.903, SD 0.098). Patients with osteoarthritis (OA) and spondyloarthropathies (SPA) reported the poorest HRQoL. In patients with RA and reactive arthritis (ReA) the HRQoL improved in a statistically significant manner during the 8-month follow-up. In addition, a clinically important change appeared in patients with systemic rheumatic diseases. HAQ score improved significantly in patients with RA, arthralgia and fibromyalgia, and ReA. In a study of 97 RA patients treated either with etanercept or adalimumab, we assessed their HRQoL with the RAND 36-Item Health Survey 1.0 (RAND-36) questionnaire. We also analysed changes in clinical parameters and the HAQ. With etanercept and adalimumab, the values of all domains in the RAND-36 questionnaire increased during the first 3 months. The efficacy of each in improving HRQoL was statistically significant, and the drug effects were comparable. Compared to Finnish age- and sex-matched general population values, the HRQoL of the RA patients was significantly lower at baseline and, despite the improvement, remained lower also at follow-up. Our RA patients had long-standing and severe disease that can explain the low HRQoL also at follow-up. In a pharmacoeconomic study of patients treated with infliximab we evaluated medical and work disability costs for patients with chronic inflammatory joint disease during one year before and one year after institution of infliximab treatment. Clinical and economic data for 96 patients with different arthritis diagnoses showed, in all patients, significantly improved clinical and laboratory variables. However, the medical costs increased significantly during the second period by 12 015 (95% confidence interval, 6 496 to 18 076). Only a minimal decrease in work disability costs occurred mean decrease 130 (-1 268 to 1 072). In a study involving a switch from infliximab to etanercept, we investigated the clinical outcome in 49 patients with RA. Reasons for switching were in 42% failure to respond by American College of Rheumatology (ACR) 50% criteria; in 12% adverse event; and in 46% non-medical reasons although the patients had responded to infliximab. The Disease Activity Score with 28 joints examined (DAS28) allowed us to measure patients disease activity and compare outcome between groups based on the reason for switching. In the patients in whom infliximab was switched to etanercept for nonmedical reasons, etanercept continued to suppress disease activity effectively, and 1-year drug survival for etanercept was 77% (95% CI, 62 to 97). In patients in the infliximab failure and adverse event groups, DAS28 values improved significantly during etanercept therapy. However, the 1-year drug survival of etanercept was only 43% (95% CI, 26 to 70) and 50% (95% CI, 33 to 100), respectively. Although the HRQoL of patients with inflammatory joint diseases is significantly lower than that of the general population, use of early and aggressive treatment strategies including TNF-inhibitors can improve patients HRQoL effectively. Further research is needed in finding new treatment strategies for those patients who fail to respond or lose their response to TNF-inhibitors.