Enterohemorrhagic Escherichia coli (EHEC) utilizes a type III secretion system (T3SS) to inject effector proteins into host cells. The EHEC NleH1 effector inhibits the nuclear factor kappa-light-chain-enhancer of activated B cells (NF-κB) pathway by reducing the nuclear translocation of the ribosomal protein S3 (RPS3). NleH1 prevents RPS3 phosphorylation by the IκB kinase-β (IKKβ). IKKβ is a central kinase in the NF-κB pathway, yet NleH1 only restricts the phosphorylation of a subset of the IKKβ substrates. We hypothesized that a protein cofactor might dictate this inhibitory specificity. We determined that heat shock protein 90 (Hsp90) interacts with both IKKβ and NleH1 and that inhibiting Hsp90 activity reduces RPS3 nuclear translocation. Full article

Many Gram-negative bacterial pathogens use type III secretion systems to deliver virulence proteins (effectors) into host cells to counteract innate immunity. The ribosomal protein S3 (RPS3) guides NF-κB subunits to specific κB sites and plays an important role in the innate response to bacterial infection. Two E. coli effectors inhibit RPS3 nuclear translocation. NleH1 inhibits RPS3 phosphorylation by IKK-β, an essential aspect of the RPS3 nuclear translocation process. NleC proteolysis of p65 generates an N-terminal p65 fragment that competes for full-length p65 binding to RPS3, thus also inhibiting RPS3 nuclear translocation. Thus, E. coli has multiple mechanisms by which to block RPS3-mediated transcriptional activation. With this in mind, we considered whether other enteric pathogens also encode T3SS effectors that impact this important host regulatory pathway. Here we report that the Salmonella Secreted Effector L (SseL), which was previously shown to function as a deubiquitinase and inhibit NF-κB signaling, also inhibits RPS3 nuclear translocation by deubiquitinating this important host transcriptional co-factor. RPS3 deubiquitination by SseL was restricted to K63-linkages and mutating the active-site cysteine of SseL abolished its ability to deubiquitinate and subsequently inhibit RPS3 nuclear translocation. Thus, Salmonella also encodes at least one T3SS effector that alters RPS3 activities in the host nucleus. Full article

Anthurium blight, caused by Xanthomonas axonopodis pv. dieffenbachiae (Xad), is one of the most serious diseases of Anthurium andraeanum. However, little is known about variations in virulence between Xad pathotypes. Here, we examined the virulence of 68 Xad strains isolated from 30 anthurium plants from five regions of China against five different anthurium cultivars. Seven bacterial pathotypes were identified based on disease index and incidence analyses following foliar spray or leaf-clip inoculation. The resulting disease susceptibility patterns for pathotypes I–VII were RRRSS, RRSRS, RSRSR, RRSSS, RSSRS, RSSSS, and SSSSS, respectively. Overall, 72% of tested strains belonged to pathotypes VI or VII and were highly virulent. A further 22.1% of strains showed medium-level virulence and were classed as pathotype III, IV, or V, while the remaining 5.9% of strains were pathotype I or II, showing low virulence. Further analysis revealed differences in the virulence of Xad strains from the same anthurium cultivar, with variation also observed in pathovars associated with the same cultivar from different areas. Our results reveal the diversity and complexity of the Xad population structure in China and suggest that investigation of Xad pathotypes provides useful information to guide the identification and use of resistant varieties of A. andraeanum. Full article

In 2017, the global Coalition for Epidemic Preparedness (CEPI) declared Lassa virus disease to be one of the world’s foremost biothreats. In January 2018, World Health Organization experts met to address the Lassa biothreat. It was commonly recognized that the diversity of Lassa virus (LASV) isolated from West African patient samples was far greater than that of the Ebola isolates from the West African epidemic of 2013–2016. Thus, vaccines produced against Lassa virus disease face the added challenge that they must be broadly-protective against a wide variety of LASV. In this review, we discuss what is known about the immune response to Lassa infection. We also discuss the approaches used to make broadly-protective influenza vaccines and how they could be applied to developing broad vaccine coverage against LASV disease. Recent advances in AIDS research are also potentially applicable to the design of broadly-protective medical countermeasures against LASV disease. Full article

This review considers current knowledge surrounding species boundaries of the Armillaria root-rot pathogens and their distribution. In addition, a phylogenetic tree using translation elongation factor subunit 1-alpha (tef-1α) from isolates across the globe are used to present a global phylogenetic framework for the genus. Defining species boundaries based on DNA sequence-inferred phylogenies has been a central focus of contemporary mycology. The results of such studies have in many cases resolved the biogeographic history of species, mechanisms involved in dispersal, the taxonomy of species and how certain phenotypic characteristics have evolved throughout lineage diversification. Such advances have also occurred in the case of Armillaria spp. that include important causal agents of tree root rots. This commenced with the first phylogeny for Armillaria that was based on IGS-1 (intergenic spacer region one) DNA sequence data, published in 1992. Since then phylogenies were produced using alternative loci, either as single gene phylogenies or based on concatenated data. Collectively these phylogenies revealed species clusters in Armillaria linked to their geographic distributions and importantly species complexes that warrant further research. Full article

We characterized a number of clinical strains of Staphylococcus spp. and investigated their sensitivity against polyphenols-rich extracts from natural raw and roasted pistachios (NPRE and RPRE, respectively). Out of 31 clinical isolates of Staphylococcus spp., 23 were coagulase-positive and identified as S. aureus, of which 21 were MRSA. Polyphenols-rich extracts from natural pistachios and roasted pistachios were prepared: the total phenols content, expressed as gallic acid equivalent (GAE)/100 g fresh weight (FW), was higher in natural pistachios (359.04 ± 8.124 mg) than roasted pistachios (225.18 ± 5.055 mg). The higher total phenols content in natural pistachios also correlated to the higher free-radical scavenging activity found by DPPH assay: NPRE and RPRE showed IC₅₀ values of 0.85 (C.L. 0.725–0.976 mg mL⁻¹) and 1.15 (C.L. 0.920–1.275 mg mL⁻¹), respectively. Both NPRE and RPRE were active against S. aureus 6538P and Staph. spp. clinical isolates, with RPRE being the most active (MIC values ranging between 31.25 and 2000 μg mL⁻¹). The antimicrobial potential of pistachios could be used to identify novel treatments for S. aureus skin infections. Full article

Silks are the long threads at the tips of maize ears onto which pollen land and sperm nuclei travel long distances to fertilize egg cells, giving rise to embryos and seeds; however fungal pathogens also use this route to invade developing grain, causing damaging ear rots with dangerous mycotoxins. This review highlights the importance of silks as the direct highways by which globally important fungal pathogens enter maize kernels. First, the most important silk-entering fungal pathogens in maize are reviewed, including Fusarium graminearum, Fusarium verticillioides, and Aspergillus flavus, and their mycotoxins. Next, we compare the different modes used by each fungal pathogen to invade the silks, including susceptible time intervals and the effects of pollination. Innate silk defences and current strategies to protect silks from ear rot pathogens are reviewed, and future protective strategies and silk-based research are proposed. There is a particular gap in knowledge of how to improve silk health and defences around the time of pollination, and a need for protective silk sprays or other technologies. It is hoped that this review will stimulate innovations in breeding, inputs, and techniques to help growers protect silks, which are expected to become more vulnerable to pathogens due to climate change. Full article

Listeria monocytogenes is a foodborne pathogen that causes listeriosis, a relatively rare, but potentially fatal, disease, with a mortality rate of 20–30%. In general, European Regulations require the absence of L. monocytogenes in five samples of 25 g before the food has left the producer, but if the food has been demonstrated not to support the growth of L. monocytogenes, up to 100 cfu g⁻¹ are allowed in the food (except for foods for infants or medical purposes) during its shelf-life under reasonably foreseeable storage conditions. It is important for food producers to determine if their food supports the growth of L. monocytogenes. The European Union Reference Laboratory for L. monocytogenes published a Technical Guidance document for conducting shelf-life studies on L. monocytogenes in ready-to-eat foods in June 2014. Primarily based on the EURL guidance document for conducting challenge studies, the ability of cheese (feta and soft goat’s milk cheese), cold-smoked salmon, coleslaw, and pork pate to support the growth of L. monocytogenes was determined using a starting inoculum of approximately 100 cfu g⁻¹. The cheese and pork pate were incubated at 8 °C for 14 days; the smoked salmon was incubated at 6 °C for 5 days and 8 °C for 9 days; and the coleslaw was incubated at 8 °C for 7 days and 12 °C for 14 days. The results showed that the smoked salmon and pork pate supported growth, while coleslaw and cheese did not. From this study, it is evident that there are factors in food other than pH, water activity, and total bacterial count (TBC) that can inhibit the ability of L. monocytogenes to grow in food. Full article

Cetylpyridinium chloride (CPC) is widely used to decontaminate water samples for the cultivation of nontuberculous mycobacteria (NTM). The rationale for using CPC is that it kills more non mycobacteria than NTM and thereby prevents the outgrowth and detection of mycobacterial colonies on solid media. The few CPC-susceptibility measurements that have been published, suggest that CPC-decontamination does kill significant numbers of NTM. We confirm that observation here and further demonstrate that CPC-susceptibility varied significantly by one log between representative NTM species and between strains of the same species. CPC-susceptibility was the same for cells collected from cultures or water-acclimated (P = 0.6485, T-test) and CPC-susceptibility was relatively similar over the range of commonly employed CPC dosages. We conclude that use of CPC as decontaminating agent may lead to failure to recover an NTM isolate and considerable underestimates of NTM numbers. Full article

Dengue virus (DENV) results in 100 million cases of infections and 22,000 deaths per year. Liver involvement, thrombocytopenia, haemorrhage and plasma leakage are characteristic manifestations of severe forms of DENV infection. However, the molecular pathways of DENV infection have not been comprehensively studied compared to the host immunological responses. We performed an in vivo study using the BALB/c mouse model with a modified mRNA differential display methodology (GeneFishing^TM) using the annealing control primer (ACP) system to capture differentially expressed genes (DEGs) in mice liver upon primary infection with DENV1 and sequential heterologous infection with DENV2. Secondary heterologous infection with DENV2 was carried out at Immunoglobulin IgM and IgG peaks following the primary DENV1 infection with the hope of determining any potential effect antibodies IgM and IgG may have on sequential heterologous infection. 30 DEGs were identified and sequenced across all three treatment groups and they belong to a variety of important pathways such as apoptosis, innate immune response, inflammatory response, metabolic processes and oxidative stress. Analysis of differentially expressed genes in response to viral infection offers valuable knowledge about the dynamic and complex association between host cell and the virus. Furthermore, some DEGs identified support DENV induced liver damage. Full article

Leishmaniasis is a neglected tropical disease with 1.6 million new cases reported each year. However, there are few safe, effective, and affordable treatments provided to those affected by this disease. Still under-appreciated as potential pharmaceutical targets, especially for cutaneous leishmaniasis infections, are the two isozymes of secreted acid phosphatase (SAP). These enzymes are involved in the survival of the parasite in the sand fly vector, and in infecting host macrophages. While the application of electric or electromagnetic fields as a medicinal therapeutic is not new, the utility of electric field application for the treatment of leishmaniasis is under studied. Studies involving the effects of electric fields on the cell secretion of SAP or the activity of SAP that has been secreted prior to electrical stimulation have not yet been reported. This work is the first report on the effect of specific electric fields on the activity of Leishmania tarentolae secreted acid phosphatases and the modulation of this secretion from the cells. In addition, the kinetic constants for the enzyme isoforms were determined as a function of days in culture and removal of carbohydrate from the glycosylated enzymes, while using a glycosidase, was shown to affect these kinetic constants. Full article

A major problem in fruit cultivation in Flanders is replant disease due to a lack of uncultivated soils available for new plantings. Replant disease can cause poor growth and affect time to full production, however Arbuscular Mycorrhizal Fungi (AMF) can prove their usefulness with regard to these problems. To further investigate the effect of AMF on nematodes, different AMF species were amended to potted apple seedlings in the presence of the nematode Pratylenchus penetrans. Generally, apple seedlings grew better in the presence of nematodes when mycorrhiza were inoculated into the soil. Moreover, a positive correlation (R² ≥ 0.88) was found between the percentage root length colonization of the roots of apple seedlings, by AMF species, and nematode reduction in the soil of the seedlings. Indigenous AMF could colonize the roots of apple seedlings the most efficiently, resulting in a higher biocontrol effect. Besides, a synergistic effect was observed when two AMF strains were applied together leading to a significant growth response of the seedlings. Full article