Healthy Host Research Articles

Endophytic Fungal Terpenoids: Natural Role and Bioactivities.

Endophytic fungi are a highly diverse group of fungi that intermittently colonize all plants without causing symptoms of the disease. They sense and respond to physiological and environmental changes of their host plant and microbiome. The inter-organism interactions are largely driven by chemical networks mediated by specialized metabolites. The balance of these complex interactions leads to healthy and strong host plants. Endophytic strains have particular machinery to produce a plethora of secondary metabolites with a variety of bioactivities and unknown functions in an ecological niche. Terpenoids play a key role in endophytism and represent an important source of bioactive molecules for human health and agriculture. In this review, we describe the role of endophytic fungi in plant health, fungal terpenoids in multiple interactions, and bioactive fungal terpenoids recently reported from endophytes, mainly from plants used in traditional medicine, as well as from algae and mangroves. Additionally, we highlight endophytic fungi as producers of important chemotherapeutic terpenoids, initially discovered in plants. Despite advances in understanding endophytism, we still have much to learn in this field. The study of the role, the evolution of interactions of endophytic fungi and their terpenoids provide an opportunity for better applications in human health and agriculture.

Gnotobiotic Plant Systems for Reconstitution and Functional Studies of the Root Microbiota.

Healthy plants host a multi-kingdom community of microbes, which is known as the plant microbiota. Amplicon sequencing technologies for microbial genomic markers were a milestone in revealing the taxonomic composition of the microbiota and its variation associated with a plant host in natural environments. However, this method alone does not allow conclusions to be drawn about functions of these microbial assemblages for the plant. The development of culture collections, which recapitulate natural microbial communities in their diversity, and multiple gnotobiotic plant systems therefore represent a breakthrough in plant-microbiota research such that plants can be inoculated with defined communities to study proposed microbiota functions. These systems provided, for the root microbiota, first insights into mechanisms underlying microbial community establishment and contributions of its microbial members to indirect pathogen protection and mineral nutrition of the host. We argue that the choice of a gnotobiotic system for microbiota reconstitution and subsequent functional analysis depends on the particular plant trait that is influenced by the microbiota. We start by discussing the advantages and limitations of using individual gnotobiotic systems and then describe the general procedures for preparing bacterial cultures from the Arabidopsis thaliana At-R-SPHERE culture collection for inoculation and cocultivation in two gnotobiotic plant growth systems using agar and perlite matrix. Additionally, a protocol for inoculation of plants with opportunistic Pseudomonas pathogens is provided. Lastly, we describe a high-throughput system for visual assessment of roots after inoculation with individual mutants of a transposon library generated from a root-derived bacterial commensal. © 2022 The Authors. Current Protocols published by Wiley Periodicals LLC. Basic Protocol 1: Preparation of bacterial cultures from At-R-SPHERE Support Protocol 1: Validation of strains by sequencing hypervariable regions of the 16S rRNA gene Basic Protocol 2: Coinoculation of plants grown on an agar matrix with microbial elicitor and a defined microbial community Alternate Protocol: Inoculation of plants cultivated in a perlite-based growth system Support Protocol 2: Surface sterilization of Arabidopsis thaliana seeds Basic Protocol 3: Inoculation using a Pseudomonas opportunistic pathogen Basic Protocol 4: Assessment of commensal-mediated root phenotypes using phytostrips.

Inhibitory Effects of Blue Light-Emitting Diode Irradiation on Podosphaera xanthii Conidial Release and Infection of Melon Seedlings

Powdery mildew fungi infect plant leaves, reducing the yield of infected melon plants. Therefore, an eco-friendly method of controlling powdery mildew in melon plants needs to be developed. A previous study described how the morphological characteristics of the conidiophores of the melon powdery mildew fungus Podosphaera xanthii Pollacci (designated KMP-6N) grown under greenhouse (natural) conditions and red light-emitting diode (LED) irradiation differed from those grown under growth chamber conditions and blue LED irradiation. In the present study, conidiophores with unconstricted conidia under blue light were collected and inoculated onto host leaves through micromanipulation; the unconstricted conidia germinated and infected the leaves, producing vigorously elongated hyphae. The number of conidia collected, the initial times of conidial release from single colonies, and the number of conidia remaining in chains on conidiophores were examined with electrostatic techniques. Under red light, the number of collected conidia gradually increased with the light irradiation period. The initial conidial release occurred between 2 to 4 h; the number of conidia remaining on the conidiophores gradually decreased and, eventually, the conidiophore lengths became shorter. In contrast, under blue light, few conidia were collected at any given time; the number of conidia on the conidiophores gradually increased and, eventually, the conidiophore lengths became longer. Next, the effects of red and blue light on the spread of powdery mildew infection by placing a KMP-6N-infected melon seedling at the centre of a tray containing healthy melon seedlings were examined. Almost all healthy seedlings caused powdery mildew symptoms at ca. 21 days after red light irradiation, whereas only healthy seedlings near the infected seedlings showed symptoms after blue light irradiation. Thus, the spread of melon powdery mildew infection clearly differed between red and blue light irradiation. This is the first report describing the effects of red and blue light on the spread of P. xanthii infection from a single infected seedling to healthy host seedlings; their results provide insight into the ecological mechanisms of powdery mildew conidial scatter from conidiophores.

Candida albicans empyema in COVID-19 infected patient: The first reported case in Qatar

Background: COVID-19 infection is caused by a novel coronavirus. It was identified initially in late 2019 as the cause of a cluster of pneumonia cases in Wuhan, China. It spread worldwide causing a pandemic. Since the start of the COVID-19 pandemic, multiple complications have been linked to COVID-19 infections, one of them being pleural effusion. To the best of our knowledge, only a few case reports have discussed the incidence of candida empyema in COVID-19 patients1,2. Case Presentation: This work is reporting on a case of COVID-19 pneumonia, in a 52-year-old previously healthy male patient, who developed ARDS 2 days after diagnosis of COVID-19 pneumonia and was attached to mechanical ventilation. Unfortunately, he continued to progress with refractory hypoxemia despite the maximum support. Ultimately, the patient required veno-venous ECMO with ultra-protective lung ventilation until the gas exchange in his lungs has normalized. The patient received multiple treatments based on our local protocol that have immunosuppressive effects. Due to worsening pneumonia, bronchoscopy was done. The Bronchoalveolar lavage (BAL) showed Candida Albicans, which was complicated with left-sided candida empyema. The later was treated by antifungal, chest tube insertion, and finally with decortication. However, despite the maximum interventions, the patient has unfortunately passed away 2 weeks after the decortication due to sepsis and multi-organ failure, while he was on ECMO during the whole period. Recommendation: Given the immunosuppressive effect of many of the medications used for the treatment of COVID-19 infection, atypical and opportunistic infections should be carefully considered within the clinical context. Conclusion: Candida pneumonia/empyema is not very common in a healthy host, however, it is a serious and rare condition, commonly seen in immunocompromised patients and associated with a high mortality rate3. The immunosuppressive therapies that are used in COVID-19 are not completely safe and more atypical infectious agents have been considered as not suitable in several typical locations causing adverse outcomes.

Ceftazidime-avibactam based combinations against carbapenemase producing Klebsiella pneumoniae harboring hypervirulence plasmids

The combination of carbapenem resistance and hypervirulence in Klebsiella pneumoniae is an emerging and urgent threat due to its potential to resist common antibiotics and cause life-threatening infections in healthy hosts. This study aimed to evaluate the activity of clinically relevant antibiotic regimens against carbapenem-resistant K. pneumoniae with hypervirulence plasmids and to identify pathways associated with antibiotic tolerance using transcriptomics. We studied two carbapenem-resistant K. pneumoniae isolates, CDI694 and CDI231, both harboring hypervirulence plasmids. Time-kill and dynamic one-compartment pharmacokinetic/pharmacodynamic assays were used to assess ceftazidime/avibactam-based therapies. RNAseq was performed following 48 h of antibiotic exposure. Closed genomes of CDI694 and CDI231 were obtained; each isolate harbored carbapenem-resistance and hypervirulence (containing rmpA/rmpA2 and iut genes) plasmids. Ceftazidime/avibactam-based regimens were bactericidal, though both isolates continued to grow in the presence of antibiotics despite no shifts in MIC. Transcriptomic analyses suggested that perturbations to cell respiration, carbohydrate transport, and stress-response pathways contributed to the antibiotic tolerance in CDI231. Genes associated with hypervirulence and antibiotic resistance were not strongly impacted by drug exposure except for ompW, which was significantly downregulated. Treatment of carbapenem-resistant K. pneumoniae harboring hypervirulence plasmids with ceftazidime/avibactam-based regimens may yield a tolerant population due to altered transcription of multiple key pathways.

Progression of COVID-19 in six South American countries with different vaccination coverage

COVID-19 seems to have progressed under two general assumptions: a) every new variant of SARS-Co V-2 should bear the worst consequences, thus justifying extension of government-mandated lock-downs, school and border closures, face mask wearing and other restrictive measures and b) that hastily developed experimental vaccines employing novel technologies would quickly and efficiently stop the pandemic by protecting from infection or, at the very least, by preventing complications and death. We tested these two hypotheses by analyzing the epidemiological data officially released by the governments of six countries in South America. We found that vaccination failed to prevent contagion by Omicron or by previous variants of SARS-Co V-2. Mortality has decreased with time since the start of global COVID-19 in early 2020 with Omicron showing the lowest mortality to date. This finding cursorily agreed with a beneficial effect of vaccination in each individual country but failed when countries with similar mortalities and different vaccination rates were analyzed. Thus, the slight but continued reduction in mortality through the duration of the pandemic and sharp decrease with Omicron do not correlate with the level of national vaccination among the countries studied. A more plausible explanation for the decrease in mortality through time is the natural attenuation of SARS-Co V-2 from successive passage through millions of susceptible and healthy hosts, as dictated by well-established principles of virology, immunology, and vaccinology dating at least from the mid 1950’s. The failure to prevent infection or to substantially reduce mortality as reported here should raise questions about the value of the massive vaccination campaigns in South America, particularly, considering the potential risks of adverse reactions (still to be fully determined) to experimental vaccines and the considerable cost to the stressed national economies of the region.

Review on Immunity to Fungal Infections in Animals

The occurrence of mycosis is relatively uncommon in healthy and immunocompetent hosts. But now a days, the incidence of fungal infections are increasing and there is no effective vaccine for fungal infections in contrast to bacterial and viral diseases. And also, available antifungal drugs are not effective to treat infected animals. Understanding the immunity against fungal infections is of interest which can contribute more for therapeutic and vaccine development. Therefore, this review focuses on the immune components involved in clearing fungal pathogens. Disease outcome is a result of host-pathogen interactions. Immunity is the body’s resistance to infection. Innate and acquired immune systems are involved to eliminate animal fungal infections. Innate immunity is not specific. It is the first line of defense, with genetically encoded receptors that identify greatly conserved pathogen-associated molecular patterns. Physical barriers, phagocytic cells, chemotactic factors and natural killer cells are some of the innate defense mechanisms. Adaptive immunity is specific. Lymphocytes have a unique and specific antigen receptor. It can be a humoral or cellular type of immune system. In adaptive immunity, there is a development of immunological memory in the host after exposure to a pathogen. However, there are no effective vaccines and antifungal drugs. So it causes high morbidity and mortality in animals and fungal pathogens have become a significant clinical challenge, leading to a global threat to controlling fungal infections. Therefore, good management of animals and treating concurrent infections strengthens their immunity. Besides, promoting research into fungal infections to develop new diagnostics, anti-fungal drugs and vaccines are recommended.

Immunomodulation and Anticancer Immunity: Reviewing the Potential of Probiotics and Their Delivery with Macromolecular Carriers.

Probiotics colonize in the gastrointestinal tract and regulate the homeostasis in healthy human hosts. These protect the host against putrefactive organisms and secrete soluble factors exhibiting important transductive roles. However, constitutive processes in human host are deregulated following dysbiosis caused during prolonged exposure to cytotoxic agents and pollutants. Apart from restoring the homeostasis, probiotic administration has shown to minimize carcinogenesis and post-surgery complications in cancer patients. Moreover, ability of microbial cells to colonize at tumor foci can be harnessed to deliver genes, therapeutic proteins and antibodies in a selective manner. In this review, we have discussed immunomodulatory roles of probiotics in context to cancer prevention. The article further proposes the use of dietary interventions for boosting anticancer immunity and as an alternative to detrimental chemotherapeutic agents. After summarizing clinical evidences on probiotic efficacy, formulation approaches have been described for effective delivery of the microorganisms. The literature shows that polysaccharide matrices can be employed to achieve the survivability of probiotics. Formulation approaches have been reviewed together with the risks associated with the migration of live microorganisms to systemic circulation and their ability of transmitting antibiotic resistance factors into human pathogens.

Time-Fractal Modulation—Possible Modulation Effects in Human Therapy

The malignant processes deviate from the healthy homeostatic control, and various “tricks” enable malignant cells to avoid the healthy regulation. Consequently, the malignant structures miss the apoptosis and proliferate without restriction, and without the formation of communication networks in the newly formed cells. The modulation supports the homeostatic control to rearrange the health regulation processes in various ways. The modulation acts with stochastic processes, using stochastic resonances for molecular excitations, supporting the regulative enzymatic processes. The number of stochastic resonant frequencies is as many as the number of enzymatic reactions. The malignant cells differ structurally and dynamically in their connections and interactions from their healthy host tissues. The radiofrequency carrier is modulated with an appropriate time-fractal (1/f) noise to select the autonomic cancer-cells, destroy them, or force the precancerous, semi-individual cells to participate in the networking connections. The modulation in this way limits the cellular autonomy of malignant cells and boosts the healthy control. The resonant energy triggers apoptotic processes and helps immunogenic actions deliver extracellular genetic information for antigen-presentation. The modulation is applied in clinical practice. The therapy (modulated electro-hyperthermia, mEHT) is intensively used in oncology in complementary applications and for palliative stages, and occasionally even as a monotherapy.

Rothia Aeria Endocarditis Complicated with Multiple Systemic Embolisms.

Rothia aeria is part of the normal flora in the human oral cavity and rarely causes serious systemic infection in healthy hosts. We report a case of infective endocarditis of the mitral valve due to Rothia aeria. A 53-year-old man suffered a cut on his left thumb. At the time, the patient licked the wound as a conventional way to accelerate its cure. Thereafter, he developed a recurrent fever, which was temporarily lysed with treatment using an intravenous antibiotic, over a period of 2 months after the injury. On admission, the patient had no dental caries and denied any dental procedures before onset of the fever. Auscultation revealed a systolic cardiac murmur. Echocardiography showed torn chordae of the posterior mitral leaflet with a small vegetation and severe mitral regurgitation. Two sets of blood cultures were positive for Rothia aeria. Computed tomography revealed splenic and left renal infarctions but no cerebral infarction. After resolution of the inflammation by 6 weeks of penicillin treatment, mitral valve repair was successfully performed.

Dietary Supplementation throughout Life with Non-Digestible Oligosaccharides and/or n-3 Poly-Unsaturated Fatty Acids in Healthy Mice Modulates the Gut-Immune System-Brain Axis.

The composition and activity of the intestinal microbial community structures can be beneficially modulated by nutritional components such as non-digestible oligosaccharides and omega-3 poly-unsaturated fatty acids (n-3 PUFAs). These components affect immune function, brain development and behaviour. We investigated the additive effect of a dietary combination of scGOS:lcFOS and n-3 PUFAs on caecal content microbial community structures and development of the immune system, brain and behaviour from day of birth to early adulthood in healthy mice. Male BALB/cByJ mice received a control or enriched diet with a combination of scGOS:lcFOS (9:1) and 6% tuna oil (n-3 PUFAs) or individually scGOS:lcFOS (9:1) or 6% tuna oil (n-3 PUFAs). Behaviour, caecal content microbiota composition, short-chain fatty acid levels, brain monoamine levels, enterochromaffin cells and immune parameters in the mesenteric lymph nodes (MLN) and spleen were assessed. Caecal content microbial community structures displayed differences between the control and dietary groups, and between the dietary groups. Compared to control diet, the scGOS:lcFOS and combination diets increased caecal saccharolytic fermentation activity. The diets enhanced the number of enterochromaffin cells. The combination diet had no effects on the immune cells. Although the dietary effect on behaviour was limited, serotonin and serotonin metabolite levels in the amygdala were increased in the combination diet group. The combination and individual interventions affected caecal content microbial profiles, but had limited effects on behaviour and the immune system. No apparent additive effect was observed when scGOS:lcFOS and n-3 PUFAs were combined. The results suggest that scGOS:lcFOS and n-3 PUFAs together create a balance—the best of both in a healthy host.

The Darkest Place Is under the Candlestick-Healthy Urogenital Tract as a Source of Worldwide Disseminated Extraintestinal Pathogenic Escherichia coli Lineages.

Since the discovery of the urinary microbiome, including the identification of Escherichia coli in healthy hosts, its involvement in UTI development has been a subject of high interest. We explored the population diversity and antimicrobial resistance of E. coli (n = 22) in the urogenital microbiome of ten asymptomatic women (representing 50% of the sample tested). We evaluated their genomic relationship with extraintestinal pathogenic E. coli (ExPEC) strains from healthy and diseased hosts, including the ST131 lineage. E. coli prevalence was higher in vaginal samples than in urine samples, and occasionally different lineages were observed in the same individual. Furthermore, B2 was the most frequent phylogenetic group, with the most strains classified as ExPEC. Resistance to antibiotics of therapeutic relevance (e.g., amoxicillin-clavulanate conferred by blaTEM-30) was observed in ExPEC widespread lineages sequence types (ST) 127, ST131, and ST73 and ST95 clonal complexes. Phylogenomics of ST131 and other ExPEC lineages revealed close relatedness with strains from gastrointestinal tract and diseased host. These findings demonstrate that healthy urogenital microbiome is a source of potentially pathogenic and antibiotic resistant E. coli strains, including those causing UTI, e.g., ST131. Importantly, diverse E. coli lineages can be observed per individual and urogenital sample type which is relevant for future studies screening for this uropathogen.

First Look Into the Ambrosia Beetle-Fungus Symbiosis Present in Commercial Avocado Orchards in Michoacán, Mexico.

Most beetle-fungus symbioses do not represent a threat to agricultural and natural ecosystems; however, a few beetles are able to inoculate healthy hosts with disease-causing fungal symbionts. Here, we report the putative nutritional symbionts associated with five native species of ambrosia beetles colonizing commercial avocado trees in four locations in Michoacán. Knowing which beetles are present in the commercial orchards and the surrounding areas, as well as their fungal associates, is imperative for developing a realistic risk assessment and an effective monitoring system that allows for timely management actions. Phylogenetic analysis revealed five potentially new, previously undescribed species of Raffaelea, and three known species (R. arxi, R. brunnea, R. fusca). The genus Raffaelea was recovered from all the beetle species and across the different locations. Raffaelea lauricola (RL), which causes a deadly vascular fungal disease known as laurel wilt (LW) in Lauraceae species, including avocado, was not recovered. This study points to the imminent danger of native ambrosia beetles spreading RL if the pathogen is introduced to Mexico's avocado orchards or natural areas given that these beetles are associated with Raffaelea species and that lateral transfer of RL among ambrosia beetles in Florida suggests that the likelihood of this phenomenon increases when partners are phylogenetically close. Therefore, this study provides important information about the potential vectors of RL in Mexico and other avocado producing regions. Confirming beetle-fungal identities in these areas is especially important given the serious threat laurel wilt disease represents to the avocado industry in Mexico.

Transmission of the Invasive Pathogen Phytophthora ramorum from Symptomatic to Healthy Host Plants During a Five-Year Period in California

Reliable data on the transmission of airborne plant pathogens are crucial for the development of epidemiological models and implementation of management strategies. The short-distance vertical transmission of the forest pathogen Phytophthora ramorum from a symptomatic California bay laurel (Umbellularia californica) to healthy containerized rhododendrons (Rhododendron caucasicum × R. ponticum var. album) was monitored for five winters (2016/17 to 2020/21) in a field experiment in Northern California. Transmission events were observed during four winters at a frequency of 1 to 17 per season, but not during the extremely dry winter of 2020/21, and were positively correlated to total rainfall rates. The first leaf symptoms were detected around mid-December and reached the highest numbers in January of most years. Only limited symptom development was observed in the spring, with the last detections in May. The exposure time (the time between the first rainfall after placing a bait plant under the bay laurel and development of symptoms) varied between 3 and over 150 days, with an average between 14 and 21 days. P. ramorum was detected from water samples collected from the canopy of the symptomatic California bay laurel. No horizontal pathogen spread was detected from symptomatic to healthy rhododendrons placed at a distance of 2 to 6 m.

Isolation, Identification and Molecular Characterization of Endophytic Fungi from the leaves of Coelogyne species, and their role as an Antimicrobial agent

Endophytic fungi from Orchids believed to have an essential symbiotic relationship with the plant for both the germination of seeds and the development of young plantlets. Endophytes are microorganisms which live in the intercellular spaces of healthy host tissues without causing apparent symptoms. Endophytic fungi isolated from the medicinal plants are used for the development of drugs as they act as the source of bioactive compounds. This study has been designed to isolate the fungal endophytes from the leaves of three epiphytic orchid species (Coelogyne nervosa, A. Rich., Coelogyne mossiae, Rolf and Coelogyne cristata, Lindl) from the same genus Coelogyne, collected from Ooty flora, Coonoor. About five different endophytes were identified and their morphological characters were studied. Coelogyne cristata showed maximum colonization frequency. Among the five species, Fusarium species showed antibacterial activity against the gram-positive bacteria Bacillus Subtilis, a pathogen found in soil, water and food. Fungal genomic DNA isolated for molecular identification from the four fungal species.

The local and global dynamics model of a cancer tumor growth

We study here, spatially homogeneous mechanistic mathematical models describing the interactions between a malignant tumor and the immune system and phase-space analysis of a certain mathematical model of tumor growth with an immune responses. Mathematical modeling of this process is viewed as a potentially powerful tool in the development of improved treatment regimens. We study some features of local and global behavior the multipoint problem for the three-dimensional tumour growth model system obtained by de Pillis and Radunskaya in 2003, with dynamics described in terms of densities of three cells populations: tumor cells, healthy host cells and effector immune cells. We found sufficient conditions, under which trajectories from the positive domain of feasible multipoint initial conditions tend to one of equilibrium points. The addition of a drug term to the system can move the solution trajectory into a desirable basin of attraction. We show that the solutions of the model with a time-varying drug term approach can be evaluated in a more fruitful way and down to earth style from the point of practical importance than the solutions of the system without drug treatment, in the condition of stimulated immune processes, only.

Human iPS cell derived RPE strips for secure delivery of graft cells at a target place with minimal surgical invasion

Several clinical studies have been conducted into the practicality and safety of regenerative therapy using hESC/iPSC-retinal pigment epithelium (RPE) as a treatment for the diseases including age-related macular degeneration. These studies used either suspensions of RPE cells or an RPE cell sheet. The cells can be injected using a minimally invasive procedure but the delivery of an intended number of cells at an exact target location is difficult; cell sheets take a longer time to prepare, and the surgical procedure is invasive but can be placed at the target area. In the research reported here, we combined the advantages of the two approaches by producing a quickly formed hiPSC-RPE strip in as short as 2 days. The strip readily expanded into a monolayer sheet on the plate, and after transplantation in nude rats, it showed a potency to partly expand with the correct apical/basal polarity in vivo, although limited in expansion area in the presence of healthy host RPE. The strip could be injected into a target area in animal eyes using a 24G canula tip.

Comparison of gut viral communities in diarrhoea and healthy dairy calves.

Calf diarrhoea has been a major cause of economic losses in the global dairy industry. Many factors, including multiple pathogen infections, can directly or indirectly cause calf diarrhoea. This study compared the faecal virome between 15 healthy calves and 15 calves with diarrhoea. Significantly lower diversity of viruses was found in samples from animals with diarrhoea than those in the healthy ones, and this feature may also be related to the age of the calves. Viruses belonging to the families Astroviridae and Caliciviridae that may cause diarrhoea in dairy calves have been characterized, which revealed that reads of caliciviruses and astroviruses in diarrhoea calves were much higher than those in healthy calves. Five complete genomic sequences closely related to Smacoviridae have been identified, which may participate in the regulation of the gut virus community ecology of healthy hosts together with bacteriophages. This research provides a theoretical basis for further understanding of known or potential enteric pathogens related to calf diarrhoea.

Extended longevity of Erwinia amylovora vectored by honeybees under in vitro conditions and its capacity for dissemination

AbstractFire blight outbreaks in Korea were first reported in 2015. Regular outbreaks have occurred since, indicating a continuous cycle of the fire blight pathogen in Korea. We determined the role of Apis mellifera (honeybee) as a vector of Erwinia amylovora by verifying the following: (a) E. amylovora longevity in/on honeybees; (b) the most common body parts that carry the bacteria; (c) the rate of bacterial spread to healthy host organs; and (d) the relationship between dispersal of viable but nonculturable (VBNC) and virulent bacterial cells. E. amylovora survived for 15 days on the exterior of honeybee bodies and was most abundant on the abdomen in comparison to other areas such as the labellum, wings, and hind legs. In the digestive system of honeybees, E. amylovora survived for 7 days, and bacteria were found in faeces for 3 days after exposure. The bacteria are likely to be VBNC on honeybees. Honeybees that were contaminated with bacteria transferred E. amylovora to healthy immature apple fruit, shoots, and flowers for 10 days after exposure. E. amylovora was also transferred from inoculated plant parts to uncontaminated honeybees. In addition, bacteria moved from inoculated plant tissues to unexposed honeybees and then from these honeybees to healthy plant tissues. Therefore, E. amylovora can survive in/on honeybees for extended amounts of time, which contradicts previous reports. The bacteria moved to host tissues via honeybees, suggesting that honeybees are the vectors of E. amylovora and play a role in the development of new outbreaks of fire blight disease in the central regions of Korea.

PLK1-dependent phosphorylation restrains EBNA2 activity and lymphomagenesis in EBV-infected mice

While Epstein–Barr virus (EBV) establishes a life-long latent infection in apparently healthy human immunocompetent hosts, immunodeficient individuals are at particular risk to develop lymphoproliferative B-cell malignancies caused by EBV. A key EBV protein is the transcription factor EBV nuclear antigen 2 (EBNA2), which initiates B-cell proliferation. Here, we combine biochemical, cellular, and in vivo experiments demonstrating that the mitotic polo-like kinase 1 (PLK1) binds to EBNA2, phosphorylates its transactivation domain, and thereby inhibits its biological activity. EBNA2 mutants that impair PLK1 binding or prevent EBNA2 phosphorylation are gain-of-function mutants. They exhibit enhanced transactivation capacities, accelerate the proliferation of infected B cells, and promote the development of monoclonal B-cell lymphomas in infected mice. Thus, PLK1 coordinates the activity of EBNA2 to attenuate the risk of tumor incidences in favor of the establishment of latency in the infected but healthy host.