Rapid Global Expansion of the Fungal Disease Chytridiomycosis into Declining and Healthy Amphibian Populations.

The fungal disease chytridiomycosis, caused by Batrachochytrium dendrobatidis, is enigmatic because it occurs globally in both declining and apparently healthy (non-declining) amphibian populations. This distribution has fueled debate concerning whether, in sites where it has recently been found, the pathogen was introduced or is endemic. In this study, we addressed the molecular population genetics of a global collection of fungal strains from both declining and healthy amphibian populations using DNA sequence variation from 17 nuclear loci and a large fragment from the mitochondrial genome. We found a low rate of DNA polymorphism, with only two sequence alleles detected at each locus, but a high diversity of diploid genotypes. Half of the loci displayed an excess of heterozygous genotypes, consistent with a primarily clonal mode of reproduction. Despite the absence of obvious sex, genotypic diversity was high (44 unique genotypes out of 59 strains). We provide evidence that the observed genotypic variation can be generated by loss of heterozygosity through mitotic recombination. One strain isolated from a bullfrog possessed as much allelic diversity as the entire global sample, suggesting the current epidemic can be traced back to the outbreak of a single clonal lineage. These data are consistent with the current chytridiomycosis epidemic resulting from a novel pathogen undergoing a rapid and recent range expansion. The widespread occurrence of the same lineage in both healthy and declining populations suggests that the outcome of the disease is contingent on environmental factors and host resistance.

Burden of fungal disease in Ireland

Our objective was to estimate the burden of fungal disease on the island of Ireland, as part of a coordinated project estimating the global burden. Published epidemiology data describing fungal infection in Ireland were identified. Population and underlying disease data were collected for 2010 and a structured set of assumptions were applied to estimate burden of fungal disease based on immunosuppression, chronic disease, and other demographic information indicating predisposition to fungal infection. From Ireland’s population of 6.4 million, we estimate 117 000 patients develop significant fungal disease each year. By far the most common fungal disease is recurrent Candida vaginitis, with an estimated 95 000 episodes annually (3000 per 100 000 women). Other fungal diseases which may be less well recognized are severe asthma with fungal sensitization and allergic bronchopulmonary aspergillosis, with estimated episodes per year of 11 700 and 9000, respectively (182 and 140 per 100 000 population, respectively). The model also estimates 450 episodes of invasive aspergillosis, 200 of chronic pulmonary aspergillosis, 600 of oesophageal candidiasis and 450 of candidaemia per year (7, 3, 9 and 6 episodes per 100 000 population, respectively). This is, we believe, the first attempt to estimate the burden of fungal disease in our population and provides a basis for estimating its impact on human health and resource use.

Revised definitions of invasive fungal disease from the European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group.

BACKGROUND: Invasive fungal diseases are important causes of morbidity and mortality. Clarity and uniformity in defining these infections are important factors in improving the quality of clinical studies. A standard set of definitions strengthens the consistency and reproducibility of such studies. METHODS: After the introduction of the original European Organization for Research and Treatment of Cancer/Invasive Fungal Infections Cooperative Group and the National Institute of Allergy and Infectious Diseases Mycoses Study Group (EORTC/MSG) Consensus Group definitions, advances in diagnostic technology and the recognition of areas in need of improvement led to a revision of this document. The revision process started with a meeting of participants in 2003, to decide on the process and to draft the proposal. This was followed by several rounds of consultation until a final draft was approved in 2005. This was made available for 6 months to allow public comment, and then the manuscript was prepared and approved. RESULTS: The revised definitions retain the original classifications of "proven," "probable," and "possible" invasive fungal disease, but the definition of "probable" has been expanded, whereas the scope of the category "possible" has been diminished. The category of proven invasive fungal disease can apply to any patient, regardless of whether the patient is immunocompromised, whereas the probable and possible categories are proposed for immunocompromised patients only. CONCLUSIONS: These revised definitions of invasive fungal disease are intended to advance clinical and epidemiological research and may serve as a useful model for defining other infections in high-risk patients.

Consensus guidelines for implementation of quality processes to prevent invasive fungal disease and enhanced surveillance measures during hospital building works, 2014

Healthcare-associated fungal outbreaks impose a substantial economic burden on the health system and typically result in high patient morbidity and mortality, particularly in the immunocompromised host. As the population at risk of invasive fungal infection continues to grow due to the increased burden of cancer and related factors, the need for hospitals to employ preventative measures has become increasingly important. These guidelines outline the standard quality processes hospitals need to accommodate into everyday practice and at times of healthcare-associated outbreak, including the role of antifungal stewardship programmes and best practice environmental sampling. Specific recommendations are also provided to help guide the planning and implementation of quality processes and enhanced surveillance before, during and after high-risk activities, such as hospital building works. Areas in which information is still lacking and further research is required are also highlighted.

Polymerase Chain Reaction Screening for Fungemia and/or Invasive Fungal Infections in Patients with Hematologic Malignancies

INTRODUCTION: Invasive fungal infections (IFIs) are a life-threatening complication in patients with hematologic malignancies, mainly in acute leukemia patients, following chemotherapy. IFI incidence is increasing, and associated mortality remains high due to unreliable diagnosis. Antifungal drugs are often limited by inadequate antimicrobial spectrum and side effects. Thus, the detection of circulating fungal DNA has been advocated as a rapid, more sensitive diagnostic tool. PATIENTS AND METHODS: Between June 01 and January 03, weekly blood samples (1,311) were screened from 193 patients undergoing intensive myelosuppressive or immunosuppressive therapy. IFI cases were classified according to European Organization for Research and Treatment of Cancer/Mycoses Study Group criteria. Fungal DNA was extracted from whole blood and amplified using polymerase chain reaction (PCR) published primers that bind to the conserved regions of the fungal 18S rRNA gene sequence. In our study, two or more consecutive positive samples were always associated with fungal disease. RESULTS: PCR screening predicted the development of IFI to be 17 days (median). This test had a specificity of 91.1% and a sensitivity of 75%. IFI incidence was 7.8%. DISCUSSION: Therefore, our results confirm the potential usefulness of PCR serial screening and the clinical applicability in everyday routine. PCR screening offers a noninvasive repeatable aid to the diagnosis of IFI.

Population genetic evidence that basidiospores play an important role in the disease cycle of rice-infecting populations of Rhizoctonia solani AG-1 IA in Iran

The fungus Rhizoctonia solani AG-1 IA causes sheath blight, one of the most important rice diseases worldwide. The first objective of this study was to analyse the genetic structure of R. solani AG-1 IA populations from three locations in the Iranian Caspian Sea rice agroecosystem. Three population samples of R. solani AG-1 IA isolates were obtained in 2006 from infected rice fields separated by 126-263km. Each field was sampled twice during the season: at the early booting stage and 45days later at the early mature grain stage. The genetic structure of these three populations was analysed using nine microsatellite loci. While the population genetic structure from Tonekabon and Amol indicated high gene flow, they were both differentiated from Rasht. The high gene flow between Tonekabon and Amol was probably due mainly to human-mediated movement of infested seeds. The second objective was to determine the importance of recombination. All three populations exhibited a mixed reproductive mode, including both sexual and asexual reproduction. No inbreeding was detected, suggesting that the pathogen is random mating. The third objective was to determine if genetic structure within a field changes over the course of a growing season. A decrease in the proportion of admixed genotypes from the early to the late season was detected. There was also a significant (P=0·002) increase in the proportion of loci under Hardy-Weinberg equilibrium. These two lines of evidence support the hypothesis that basidiospores can be a source of secondary inoculum. © 2012 BSPP.

The plasticity of NBS resistance genes in sorghum is driven by multiple evolutionary processes

Background Increased disease resistance is a key target of cereal breeding programs, with disease outbreaks continuing to threaten global food production, particularly in Africa. Of the disease resistance gene families, the nucleotide-binding site plus leucine-rich repeat (NBS-LRR) family is the most prevalent and ancient and is also one of the largest gene families known in plants. The sequence diversity in NBS-encoding genes was explored in sorghum, a critical food staple in Africa, with comparisons to rice and maize and with comparisons to fungal pathogen resistance QTL. Results In sorghum, NBS-encoding genes had significantly higher diversity in comparison to non NBS-encoding genes and were significantly enriched in regions of the genome under purifying and balancing selection, both through domestication and improvement. Ancestral genes, pre-dating species divergence, were more abundant in regions with signatures of selection than in regions not under selection. Sorghum NBS-encoding genes were also significantly enriched in the regions of the genome containing fungal pathogen disease resistance QTL; with the diversity of the NBS-encoding genes influenced by the type of co-locating biotic stress resistance QTL. Conclusions NBS-encoding genes are under strong selection pressure in sorghum, through the contrasting evolutionary processes of purifying and balancing selection. Such contrasting evolutionary processes have impacted ancestral genes more than species-specific genes. Fungal disease resistance hot-spots in the genome, with resistance against multiple pathogens, provides further insight into the mechanisms that cereals use in the “arms race” with rapidly evolving pathogens in addition to providing plant breeders with selection targets for fast-tracking the development of high performing varieties with more durable pathogen resistance.

Alternative fungicides for controlling husk spot caused by Pseudocercospora macadamiae in macadamia

Husk spot, caused by Pseudocercospora macadamiae is a major fungal disease of macadamia in Australia. Chemicals to control the disease are limited and frequent failure to control the disease is a major concern to growers. The overall goal of this research was to improve the chemical control strategy of P. macadamiae through the provision of fungicides with different modes of action to carbendazim, which is the current industry standard. Husk spot incidence, premature fruit abscission, kernel quality and yield were evaluated following application of different fungicide products in replicated field experiments at three different sites. Results showed significant differences in disease incidence and premature fruit abscission between fungicide treatments, field sites and years. Generally, disease incidence and premature fruit abscission on trees treated with fungicide were significantly (P < 0.05) lower than the untreated control. Pyraclostrobin conferred significantly better protection than trifloxystrobin, reducing disease severity by 70% compared with a 50% reduction by trifloxystrobin. The pyraclostrobin treatment had a similar efficacy to the current industry standard (70% reduction cf. 73% reduction by tank-mixed carbendazim and copper). Higher amounts of immature kernels occurred in the untreated control, followed by difenoconazole and trifloxystrobin. Diseased fruit accounted for 78% of premature fruit abscission, which indicates that husk spot enhances fruit abscission in macadamia. Our results suggest that pyraclostrobin provided similar efficacy to the industry standard and could, therefore, play a key role in the management of husk spot.

Evaluation of transgenic bananas expressing anti-apoptotic genes for resistance against Fusarium wilt

The banana industry worldwide is under threat from a fungal disease known as Fusarium wilt, a disease for which there is no chemical control. Conventional breeding approaches to generate resistant banana varieties are lengthy and very difficult. As such, genetic engineering for disease resistance is considered the most viable control option. In this PhD thesis, genetically modified banana plants were generated using several different stress tolerance genes. When challenged with Fusarium wilt in glasshouse trials, some lines showed increased resistance to the disease. The promising elite lines generated in this study will now require testing in field trials.

Controlling Pythium in Ginger: Phase 2

The fungal disease Pythium Soft Rot is regarded by the ginger industry as their most serious disease threat. This project was developed to further investigate the factors contributing to the persistence and spread of Pythium Soft Rot on ginger farms and to identify measures for their control. The study demonstrated that the pathogen capable of causing Pythium Soft Rot in ginger was spread in contaminated ‘seed’, soil and water and that it can be managed through a combination of strategies that rely on early detection, reducing pathogen levels in soils, preventing water logging and restricting movement of contaminated ‘seed’, soil and water. However, in order to have an effective level of control, all strategies need to be integrated in an effective manner.

The plasticity of NBS resistance genes in sorghum is driven by multiple evolutionary processes

Background Increased disease resistance is a key target of cereal breeding programs, with disease outbreaks continuing to threaten global food production, particularly in Africa. Of the disease resistance gene families, the nucleotide-binding site plus leucine-rich repeat (NBS-LRR) family is the most prevalent and ancient and is also one of the largest gene families known in plants. The sequence diversity in NBS-encoding genes was explored in sorghum, a critical food staple in Africa, with comparisons to rice and maize and with comparisons to fungal pathogen resistance QTL. Results In sorghum, NBS-encoding genes had significantly higher diversity in comparison to non NBS-encoding genes and were significantly enriched in regions of the genome under purifying and balancing selection, both through domestication and improvement. Ancestral genes, pre-dating species divergence, were more abundant in regions with signatures of selection than in regions not under selection. Sorghum NBS-encoding genes were also significantly enriched in the regions of the genome containing fungal pathogen disease resistance QTL; with the diversity of the NBS-encoding genes influenced by the type of co-locating biotic stress resistance QTL. Conclusions NBS-encoding genes are under strong selection pressure in sorghum, through the contrasting evolutionary processes of purifying and balancing selection. Such contrasting evolutionary processes have impacted ancestral genes more than species-specific genes. Fungal disease resistance hot-spots in the genome, with resistance against multiple pathogens, provides further insight into the mechanisms that cereals use in the “arms race” with rapidly evolving pathogens in addition to providing plant breeders with selection targets for fast-tracking the development of high performing varieties with more durable pathogen resistance.

Resistance of Norway spruce (Picea abies) to root and butt rot (Heterobasidion parviporum) in peatland and mineral soil

Root and butt rot is the most harmful fungal disease affecting Norway spruce in southern Finland. In approximately 90 % of cases the causal agent is Heterobasidion parviporum. Root and butt rot infections have not been reported in Finnish peatlands. However, the increase in logging operations in peatlands means there is a risk that the fungus will eventually spread to these areas. The aim of this study was to find out the impact of growing site on the resistance of Norway spruce to Heterobasidion parviporum infections. This was investigated by artificially inoculating H. parviporum to spruce trees in pristine mire, drained peatland and mineral soil and comparing the defence reactions. Additionally, the effect of genotype on resistance was studied by comparing the responses of spruce clones representing different geographic origins. The roots and stems of the trees to be sampled were wounded and inoculated with wood dowels pre-colonised by H. parviporum hyphae. The resulting necrosis around the point of inoculation was observed. It was presumed that increased length of necrosis indicates high susceptibility of the tree to the disease. The relationship between growth rate and host resistance was also studied. The results indicated that growing site does not have a statistically significant effect on host resistance. The average length of necrosis around the point of inoculation was 35 mm in pristine mire, 37 mm in drained peatland and 40 mm in mineral soil. It was observed that growth rate does not affect resistance, but that the genotype of the tree does have an effect. The most resistant spruce clone was the one with Russian origin. The results suggest that the spruce stands in peatlands are not more resistant to root and butt rot infections than those in mineral soil. These findings should be taken into consideration when logging peatland forests.

The economic impact of diseases and parasitic problems in freshwater fish production

Diseases and parasitic problems could constitute significant economic losses in fish production if not controlled, thus the need to continue monitoring its prevalence. Based on field studies on feral and intensively raised fish at the Kainji Lake Research Institute Nigeria, some diseases and parasitic problems have been identified. These include; helminthiasis; fungal disease; protozoa which include Myxosoma sp., Myxobolus spp., Henneguya sp., Trichodina sp., Ichthopthrius sp. bacterial mainly Aeromonas sp., Pseudomonas sp., mechanical injuries; death due to unknown causes and economic assessment of myxosporidian infection. Suggestion for disease control in fish production are recommended

Aphanomycosis of crayfish: crayfish plague

This is the Aphanomycosis of crayfish: crayfish plague report produced by the Environment Agency in 2000. Crayfish plague is an extremely virulent fungal disease of European crayfish species, the white clawed or stone crayfish of Western Europe Austropotamobius pallipes, the Noble crayfish of northern Europe Astacus astacus and the narrow clawed crayfish of Eastern Europe, Astacus leptodactylus. The white claw crayfish A. pallipes is the indigenous native crayfish of the British Isles. Until the early 1980s there were extensive healthy populations of this crayfish in almost all suitable alkaline river and lake environments in England and Wales as far north as Northumberland. The conservation importance of this native crayfish is widely recognised. This report provides a general review of the literature of crayfish plague, including an overview of its spread through the British Isles from CEFAS records. Information on current diagnostic methods from the Office International des Epizooties (OIE) Aquatic Disease Manual is provided. Information on the taxonomy, morphology and physiology of the pathogen is reviewed, together with the pathogenicity and pathology of the disease and current means of prevention and control.

Notes on the induced maturation and spawning in four-month-old Penaeus monodon Fabricius by eyestalk ablation

The ablation technique consisted of making an incision across the eyeball to allow free flow of fluids while holding the prawn under water, squeezing the eyeball contents outwards, and pinching hard the eyestalk tissue. The cut area heals completely in about a week; no application of antibiotics is necessary. Spent spawners were tagged with thin brass rings (Rodriguez, 1976) around the unablated eyestalk for a separate experiment on rematuration. Two spawning yielding approximately 277,000 eggs were obtained three weeks after ablation, followed four days later by two more spawnings with 160,000 eggs; all four spawners weighed more than 100 g. With a hatching rate of 98% and 78% for the first and second batch, respectively, the spawnings produced viable nauplii. Water temperatures as low as 23 degree C due to a delayed cold spell in March depressed molting; weakened larvae had to be discharged at the mysis stage. Although ovarian development continued, no further spawnings were obtained due mainly to the onset of bacterial and fungal disease. Infection is initiated in injured portions of the exoskeleton, sometimes penetrating right through the muscles to the ovarian tissues. The non-flowthrough conditions and mussel meat feeding led to fouling of the culture water resulting in consecutive mortalities caused by disease. Female P.monodon held in maturation pens were ablated at the age of 15 months (Santiago, et al., 1976); they averaged only 16 g body weight after four months growth in ponds. In another experiment, pond-reared P.monodon females ranging from 50 to 80 g were ablated at approximately seven months (Aquacop, 1977). The present results show a minimum age of four months from postlarve that P.monodon is capable of ovarian development and spawning upon ablation. However, maturation is probably affected by size as well as age - the four-month old females weighed an average of 100 g in contrast to the smaller animals in the earlier experiments.