Erwinia amylovora: the molecular basis of fireblight disease

Taxonomy: Bacteria; Proteobacteria; γ subdivision; order Enterobacteriales; family Enterobacteriaceae; genus Erwinia .
Microbiological properties: Gram-negative, motile rods.
Related species: E. carotovora (soft-rot diseases) , E. chrysanthemi (soft-rot diseases) , E. (Pantoea) stewartii (Stewart's wilt of corn) , E. (Pantoea) herbicola (epiphyte).
Host range: Affects rosaceous plants, primarily members of the Pomoideae . Economically important hosts are apple and pear. The commercial implications of fireblight outbreaks are aggravated by the limited effectiveness of current control measures.
Disease symptoms: E. amylovora infection is characterized by water soaking of infected tissue, followed by wilting and tissue necrosis. Necrosis gives tissue a scorched, blackened appearance, giving rise to the name fireblight. Symptoms are often localized to blossom bracts or young shoots but, in highly susceptible hosts, can spread systemically resulting in death of the entire tree. Infections can vary in severity depending on climatic conditions and host susceptibility.
Useful web site: http://www.agric.gov.ab.ca     

Weather and fireblight in England

A method for assessing potential fireblight activity in the field in southeast England is described. It is based on potential doublings (PD), derived from in vitro growth rates of Erwinia amylovora at different temperatures, combined with a rain score (R) derived from daily precipitation values. Where the incubation period (I) was known, the coefficient of correlation between I/R and PD was 085 (P < 0–001). When I was regressed on PD and R, the multiple correlation coefficient was 0–97. The low level of fireblight in England precluded direct testing of the predictive value of the relationship between PD, I and R but its application to growing season weather of 1955-74 highlighted those years when fireblight activity was high in a particular host. Whilst within the range 65–86 ^oF (18–30 ^oC) there is good agreement between accumulated potential doubling values and accumulated degree days, the rain score would probably require modification in other climates.     

The Amphibian Chytrid Batrachochytrium dendrobatidis Occurs on Freshwater Shrimp in Rain Forest Streams in Northern Queensland, Australia

Chytridiomycosis is a disease of amphibians caused by the chytrid fungus Batrachochytrium dendrobatidis. It can be highly virulent and is unusual in that it appears to drive many host species to local extinction during outbreaks. One mechanism that could facilitate this is the ability to grow saprophytically or on alternative hosts. This is common in other chytrids but has not been demonstrated for B. dendrobatidis in the field. B. dendrobatidis can grow on arthropod exoskeletons in the laboratory, and freshwater shrimp can be the most abundant animals in tropical rain forest streams. We therefore used diagnostic quantitative polymerase chain reaction to determine the infection status of freshwater shrimp from areas in which they are sympatric with frog species that have suffered declines in association with outbreaks of chytridiomycosis. We detected B. dendrobatidis on three individual shrimp belonging to two genera and collected from two widely separated streams. Two of the individuals had high levels of infection. This indicates that the presence of alternative hosts is likely to contribute to the extreme virulence of chytridiomycosis outbreaks in some systems. The presence of alternative hosts may allow B. dendrobatidis to remain in the environment after local extinctions of amphibian hosts, preventing the recovery of amphibian populations.     

Fireblight Erwinia amylovora and weather: a comparison of warning systems

Warning systems for fireblight Erwinia amylovora developed in New York, Illinois and California, USA, and in south-east England are compared. General principles which might be applicable in the different climates were sought. The consequences of applying threshold temperature values chosen for one area in a different climatic area were examined using Sacramento, California; Rochester, New York; Vlissingen, The Netherlands; Kent, England as examples. A graded system for assessing fireblight risks, derived from all the systems, is suggested. It takes into account both risks of infection and risks of high insect activity and it is best used in conjunction with Billing's incubation period assessment system.     

Phytotoxic material from associations betweenErwinia amylovora and pear tissue culture: Possible role in necrotic symptomatology of fireblight disease

Pear cell suspension cultures (PCSC) inoculated with virulent strains of the fireblight bacterium,Erwinia amylovora, exhibited massive necrosis within 7 days, whereas avirulentE. amylovora strains and other enterobacteria generally elicited very slight or no necrotic reactions. These results were generally repeated when pear seedlings (1–2 months old) were inoculated with these same bacterial strains. Fractions derived from massively necrotized PCSC were tested for biological activity against healthy PCSC and pear seedling cuttings. Of these fractions, the dialyzable, 70%-ethanol-soluble material—remaining after precipitation by ethanol of high-molecular-weight polysaccharides (“amylovorin”)—exclusively inhibited growth of pear callus, plasmolyzed and disintegrated pear cells in PCSC, and caused some browning of pear callus. Although cuttings of young pear seedlings placed in solutions containing this dialyzable, ethanol-soluble material generally became massively necrotized within 3 days, some residual methodological problems with this bioassay procedure must be solved. These observations suggest that some relatively small molecule(s) formed in PCSC inoculated with virulentE. amylovora can exert antagonistic biological activity against pear tissue and may play a role in the symptomatology characteristic of the fireblight disease.     

Bats and zoonotic viruses: can we confidently link bats with emerging deadly viruses?

An increasingly asked question is ''can we confidently link bats with emerging viruses?''. No, or not yet, is the qualified answer based on the evidence available. Although more than 200 viruses - some of them deadly zoonotic viruses - have been isolated from or otherwise detected in bats, the supposed connections between bats, bat viruses and human diseases have been raised more on speculation than on evidence supporting their direct or indirect roles in the epidemiology of diseases (except for rabies). However, we are convinced that the evidence points in that direction and that at some point it will be proved that bats are competent hosts for at least a few zoonotic viruses. In this review, we cover aspects of bat biology, ecology and evolution that might be relevant in medical investigations and we provide a historical synthesis of some disease outbreaks causally linked to bats. We provide evolutionary-based hypotheses to tentatively explain the viral transmission route through mammalian intermediate hosts and to explain the geographic concentration of most outbreaks, but both are no more than speculations that still require formal assessment.     

The prevalence and distribution of sheep scab in Wales: a farmer questionnaire survey

Outbreaks of ovine psoroptic mange in the U.K. have increased 100‐fold since its deregulation in 1992, with the highest prevalence in Wales, a region of high sheep density. A cross‐sectional, retrospective, questionnaire‐based survey of 7500 members of the association of Welsh lamb and beef farmers [Welsh Lamb and Beef Producers Ltd (WLBP)] was used to investigate the prevalence and distribution of sheep scab in this region in 2015. The survey was completed by 14.0% (n = 972) of potential respondents. Scab outbreaks were reported on 15.8% (n = 154) of farms in 2015. However, 29.0% (n = 282) of farms reported at least one scab outbreak and 2.4% (n = 23) of farms had experienced between six and 10 outbreaks in the previous 10 years. Most outbreaks occurred during September–January (83.0%, n = 150), and were clustered around Brecon (mid‐Wales) and Bangor (North Wales). Farmers who used common grazing were significantly more likely to report scab outbreaks in the previous 10 years than farmers who did not. No quarantine procedures for sheep bought in were used by 29.0% (n = 262) of farmers. Future research should be directed towards the development of localized management programmes, with a particular focus on areas of common grazing.     

Multiple Merger Genealogies in Outbreaks of Mycobacterium tuberculosis

The Kingman coalescent and its developments are often considered among the most important advances in population genetics of the last decades. Demographic inference based on coalescent theory has been used to reconstruct the population dynamics and evolutionary history of several species, including Mycobacterium tuberculosis (MTB), an important human pathogen causing tuberculosis. One key assumption of the Kingman coalescent is that the number of descendants of different individuals does not vary strongly, and violating this assumption could lead to severe biases caused by model misspecification. Individual lineages of MTB are expected to vary strongly in reproductive success because 1) MTB is potentially under constant selection due to the pressure of the host immune system and of antibiotic treatment, 2) MTB undergoes repeated population bottlenecks when it transmits from one host to the next, and 3) some hosts show much higher transmission rates compared with the average (superspreaders).Here, we used an approximate Bayesian computation approach to test whether multiple-merger coalescents (MMC), a class of models that allow for large variation in reproductive success among lineages, are more appropriate models to study MTB populations. We considered 11 publicly available whole-genome sequence data sets sampled from local MTB populations and outbreaks and found that MMC had a better fit compared with the Kingman coalescent for 10 of the 11 data sets. These results indicate that the null model for analyzing MTB outbreaks should be reassessed and that past findings based on the Kingman coalescent need to be revisited.     

Biology and Evolution of Beneficial and Detrimental Viruses of Animals, Plants, and Fungi

Virtually every type of organism may serve as a host for viruses. In some hosts, virus presence may be considered beneficial to humans; in other hosts, viruses are considered detrimental. Examples of viruses that are considered beneficial to humans include those that are used for biological control of organisms that themselves are considered detrimental to humans, such as plant pathogenic fungi. Viruses are extremely variable in terms of morphology, structure, and genome organization. However, viruses that attack hosts from different kingdoms may be related, deriving from the same phylogeny. This paper summarizes some of the properties of three related families of viruses that attack hosts in different kingdoms: the animal-infecting Picornaviridae, the plant-infecting Potyviridae, and the fungus-infecting Hypoviridae. Properties of these viruses that set them apart from each other and factors that may affect their evolution are discussed.     

Role Of Seedbank Substrates in the Revegetation of Fly Ash and Gypsum in the United Kingdom

Experimental plots of pulverized fuel ash (PFA), alone or mixed with flue gas desulphurization (FGD) gypsum, were seeded with topsoil from areas where PFA had been revegetated naturally, or with estuarine soil. Plots containing fresh PFA became more saline during the first three years due to formation of a salt crust, and these plots experienced colonization by halophytic species. Plots initially containing 2-year-old PFA declined in salinity throughout the experiment and were colonized by a richer plant community dominated by legumes. A total of 57 plant species was recorded in the two experiments. Of these, Melilotus officinalis (ribbed melilot), Medicago lupulina (black medick), Vulpia myuros (rat's tail fescue), and Puccinellia maritima (common saltmarsh grass) showed commercial potential for stabilizing these wastes, and by implication other saline or high-boron materials.     

Polyphagy in an uncertain environment: Helicoverpa punctigera in inland Australia

We investigated potential primary hosts for Helicoverpa punctigera, a polyphagous lepidopteran pest which affects many crops, but also has many native hosts in non‐cropping regions of inland Australia. The most common hosts are Asteraceae and Fabaceae, but there is little information on the relative importance of different species in these families for the population dynamics of H. punctigera in the inland. We tested three common hosts, one legume (Cullen cinereum) and two daisies, Rhodanthe floribunda and Polycalymma stuartii, for their suitability for survival and growth of early instar larvae. Survival was high on the flowers of all three species, and on the leaves of C. cinereum and R. floribunda, but low on leaves of P. stuartii. There were significant differences in growth rates, measured by head widths, in the order of C. cinereum > R. floribunda > P. stuartii, and flowers > leaves. They were strongly correlated with nitrogen content, measured by fresh weight. Field surveys of larval populations in western Queensland indicated that two legume species, C. cinereum and Cullen pallidum, were widely utilized and supported many larvae. Daisies were generally less widely used and/or had lower numbers of larvae. When C. cinereum or C. pallidum was found near daisies, the legume had more larvae if both the daisy and the legume were flowering, but this was reversed if only the daisy was flowering. The seasonality and habitats of the daisies and legumes are different. C. cinereum and C. pallidum (a new host record for H. punctigera) may be primary hosts of H. punctigera in inland Australia and key hosts for maintaining populations in flood plains. However the areas occupied by daisies can be more extensive depending on rainfall, and some species may be critical in generating outbreaks that affect cropping areas in southeast Australia.     

Prediction of the potential global distribution of the Asian longhorned beetle Anoplophora glabripennis (Coleoptera: Cerambycidae) under climate change

1. The Asian longhorned beetle (ALB) Anoplophora glabripennis (Motschulsky) (Coleoptera: Cerambycidae) is an important wood-boring pest that has caused substantial damage to broadleaf trees in Asia, North America, and Europe.
2. We used the modelling software CLIMEX to project the potential global distribution of ALB based on both historical (1987–2016) and future (2021–2050) climate conditions. ALB has possible hosts in 37 genera, and their known distributions were incorporated into the model to assess their effect on pest distribution.
3. Suitable regions for ALB are predicted to be widely distributed under both historical and future climate conditions, and across all continents except Antarctica. With climate change, climate suitability would increase in the regions north of 30°N and decline in most regions south of 30°N.
4. The area of most climate-suitable regions would be covered by potential hosts, and optimum hosts would dominate. The possibility of ALB outbreaks in the Northern Hemisphere is much higher than in the Southern Hemisphere, owing to the richer abundance of hosts.
5. These results provide theoretical guidance for developing effective ALB monitoring and mitigation measures.

     

Lab on a chip genotyping for Brucella spp. based on 15-loci multi locus VNTR analysis

Background  

Brucellosis is an important zoonosis caused by the genus Brucella. In addition Brucella represents potential biological warfare agents due to the high contagious rates for humans and animals. Therefore, the strain typing epidemiological tool may be crucial for tracing back source of infection in outbreaks and discriminating naturally occurring outbreaks versus bioterroristic event. A Multiple Locus Variable-number tandem repeats (VNTR) Analysis (MLVA) assay based on 15 polymorphic markers was previously described. The obtained MLVA band profiles may be resolved by techniques ranging from low cost manual agarose gels to the more expensive capillary electrophoresis sequencing. In this paper a rapid, accurate and reproducible system, based on the Lab on a chip technology was set up for Brucella spp. genotyping.     

Host plant affects pollen beetle (Meligethes aeneus) egg size

Abstract. In some herbivorous insect species, egg size is larger on low‐quality hosts than on high‐quality hosts and may be related to the prospect that larger offspring are more likely to survive on a poor host. Sizes of eggs laid by pollen beetles [Meligethes aeneus Fab. (Coleoptera: Nitidulidae)] were examined with insects confined on one of two different host plants that had previously shown differences in adult preference and larval performance. Individual females were also exposed sequentially to both the low‐quality host (Sinapis alba L.) and the high‐quality host (Brassica napus L.) and the size of their eggs was determined. Pollen beetles laid shorter eggs on low‐quality hosts both for different females on different host plants and for the same individuals on different host plants, in contrast to the prediction that low‐quality hosts would receive larger eggs than high‐quality hosts. Previously, egg production rate was shown to be reduced when pollen beetles are exposed to low‐quality hosts and it is suggested that oogenesis is incomplete, resulting in shorter eggs. The possibility that this is related to antibiosis on S. alba is discussed.     

Estimating distemper virus dynamics among wolves and grizzly bears using serology and Bayesian state‐space models

Many parasites infect multiple hosts, but estimating the transmission across host species remains a key challenge in disease ecology. We investigated the within and across host species dynamics of canine distemper virus (CDV) in grizzly bears (Ursus arctos) and wolves (Canis lupus) of the Greater Yellowstone Ecosystem (GYE). We hypothesized that grizzly bears may be more likely to be exposed to CDV during outbreaks in the wolf population because grizzly bears often displace wolves while scavenging carcasses. We used serological data collected from 1984 to 2014 in conjunction with Bayesian state‐space models to infer the temporal dynamics of CDV. These models accounted for the unknown timing of pathogen exposure, and we assessed how different testing thresholds and the potential for testing errors affected our conclusions. We identified three main CDV outbreaks (1999, 2005, and 2008) in wolves, which were more obvious when we used higher diagnostic thresholds to qualify as seropositive. There was some evidence for increased exposure rates in grizzly bears in 2005, but the magnitude of the wolf effect on bear exposures was poorly estimated and depended upon our prior distributions. Grizzly bears were exposed to CDV prior to wolf reintroduction and during time periods outside of known wolf outbreaks, thus wolves are only one of several potential routes for grizzly bear exposures. Our modeling approach accounts for several of the shortcomings of serological data and is applicable to many wildlife disease systems, but is most informative when testing intervals are short. CDV circulates in a wide range of carnivore species, but it remains unclear whether the disease persists locally within the GYE carnivore community or is periodically reintroduced from distant regions with larger host populations.     

Testing a key assumption of host‐pathogen theory: density and disease transmission

Conventional disease theory suggests that extinction with density‐dependent transmission is unlikely as the threshold host density (KT) is greater than zero. Extinction may result if transmission is frequency dependent or the pathogen has an environmental reservoir. Given the importance of understanding how pathogens affect species richness and diversity there are few empirical tests of these conclusions. We used an Ambystoma tigrinum–Ambystoma tigrinum virus (ATV) model system in the laboratory to examine disease transmission dynamics. Susceptible A. tigrinum larvae were exposed to three different densities and proportions of infected larvae for 24 h. We then housed susceptible hosts individually for 28 days and monitored them for infection. The density of infected hosts to which susceptible hosts were exposed was the best predictor of infection (p=0.037). There was no effect of host clutch on the probability of becoming infected (p=0.67). Larvae in the highest density treatments died sooner than larvae in lower density treatments (p<0.001). Asymptomatic but infected hosts shed sufficient virus into the water in a 24‐h period to infect susceptible hosts without any direct contact between individuals. ATV transmission was best described by a power function, leading to the prediction that extinction of A. tigrinum as a result of this pathogen is unlikely. Indeed, field observations show that larval salamander populations that experience ATV‐driven epidemics may decrease, but not to extinction, and then recover. Disease is proposed as a possible explanation for the global decline of amphibians. Ranaviruses infect many amphibian populations, but based on our results may not be a general cause of declines to extinction. In contrast, frequency dependent transmission, environmental reservoirs and alternative hosts may be the most likely explanation for the enigmatic decline, at times to extinction, of some amphibian populations as a result of emerging infectious diseases, like the chytrid fungus Batrachochytrium dendrobatidis.     

Olfactory recognition and behavioural avoidance of angiosperm nonhost volatiles by conifer-inhabiting bark beetles

Abstract 1 When searching for suitable hosts in flight, especially in mixed forests, conifer‐inhabiting bark beetles will encounter not only suitable host trees and their odours, but also unsuitable hosts and nonhost trees. Rejection of these trees could be based on an imbalance of certain host characteristics and/or a negative response to some nonhost stimuli, such as nonhost volatiles (NHV). 2 Recent electrophysiological and behavioural studies clearly indicate that conifer‐inhabiting bark beetles are not only able to recognize, but also to avoid, nonhost habitats or trees by olfactory means. Green leaf volatiles (GLV), especially C₆‐alcohols, from the leaves (and partly from bark) of nonhost angiosperm trees, may represent nonhost odour signals at the habitat level. Specific bark volatiles such as trans‐conophthorin, C₈‐alcohols, and some aromatic compounds, may indicate nonhosts at the tree species level. Flying bark beetles are also capable of determining whether a possible host is unsuitable by reacting to signals from conspecifics or sympatric heterospecifics that indicate old or colonized host tree individuals. 3 Combined NHV signals in blends showed both redundancy and synergism in their inhibitory effects. The coexistence of redundancy and synergism in negative NHV signals may indicate different functional levels (nonhost habitats, species, and unsuitable hosts) in the host selection process. Combinations of NHV and verbenone significantly reduced the number of mass attacked host trees or logs on several economically important species (e.g. Dendroctonus ponderosae, Ips typographus, and I. sexdentatus). 4 We suggest a semiochemical‐diversity hypothesis, based on the inhibition by NHV of bark beetle host‐location, which might partly explain the lower outbreak rates of forest insects in mixed forests. This ‘semiochemical‐diversity hypothesis’ would provide new support to the general ‘stability‐diversity hypothesis’. 5 Natural selection appears to have caused conifer‐inhabiting bark beetles to evolve several olfactory mechanisms for finding their hosts and avoiding unsuitable hosts and nonhost species. NHV and unsuitable host signals have potential for use in protecting trees from attack. The use of these signals may be facilitated by the fact that their combination has an active inhibition radius of several metres in trap test, and by the observation of area effects for several trees near inhibitor soruces in tree protection experiments. Furthermore, incorporation of negative signals (such as NHV and verbenone) and pheromone‐based mass‐trapping in a ‘push–pull’ fashion may significantly increase the options for control against outbreaks of conifer‐inhabiting bark beetles, especially in high risk areas.     

Glyoxal as an alternative fixative to formaldehyde in immunostaining and super‐resolution microscopy

Paraformaldehyde (PFA) is the most commonly used fixative for immunostaining of cells, but has been associated with various problems, ranging from loss of antigenicity to changes in morphology during fixation. We show here that the small dialdehyde glyoxal can successfully replace PFA. Despite being less toxic than PFA, and, as most aldehydes, likely usable as a fixative, glyoxal has not yet been systematically tried in modern fluorescence microscopy. Here, we tested and optimized glyoxal fixation and surprisingly found it to be more efficient than PFA‐based protocols. Glyoxal acted faster than PFA, cross‐linked proteins more effectively, and improved the preservation of cellular morphology. We validated glyoxal fixation in multiple laboratories against different PFA‐based protocols and confirmed that it enabled better immunostainings for a majority of the targets. Our data therefore support that glyoxal can be a valuable alternative to PFA for immunostaining.     

Feeding preferences and their modification in early and late instar larvae of the bagworm,Thyridopteryx ephemeraeformis (Lepidoptera: Psychidae)

Feeding preferences in naive first instar and penultimate-final instar bagworm (Thyridopteryx ephemeraeformis)larvae were investigated using two choice tests. Naive first instar larvae showed a general (though not exclusive) preference for black willow (Salix nigra)over other hosts tested, regardless of the host of origin. Penultimate-final instar larvae showed strong preferences for the previous rearing host, suggesting induction of host preference; there was no evidence of host origin effects. The ability to discriminate among hosts is important if it is actually used to make host choices based on host quality. In bagworms, where host choices (if any) must be made by larvae in habitats where several hosts are often present, host discriminatory ability is likely important.     

6-Thioguanine fromErwinia amylovora

Erwinia amylovora, the etiological agent of fireblight disease, was found to produce under aerobic conditions in axenic culture 2-amino-1,7-dihydro-6H-purine-6-thion (6-thioguanine, 6-TG), easily recognized by a UV absorbance at 340 nm (pH7) and readily distinguished from NADH by both its acid stability and the pH dependence of its UV spectrum. 6-TG is known for a variety of physiological activities, but in this study no toxicity toward Bartlett pear suspension culture was detected. Although a synergistic role of 6-TG in the development of fireblight is not excluded, it may not be responsible for the black necrosis typical for the disease. This conclusion was corroborated through the study of severalE. amylovora mutants with respect to their capability to produce 6-TG.