Genomics‐informed design of loop‐mediated isothermal amplification for detection of phytopathogenic Xanthomonas arboricola pv. pruni at the intraspecific level

The objective of this study was to develop a rapid, sensitive detection assay for the quarantine pathogen Xanthomonas arboricola pv. pruni, causal agent of stone fruit bacterial spot, an economically important disease of Prunus spp. Unique targets were identified from X. arboricola pv. pruni genomes using a comparative genomics pipeline of other Xanthomonas species, subspecies and pathovars, and used to identify specific diagnostic markers. Loop‐mediated isothermal amplification (LAMP) was then applied to these markers to provide rapid, sensitive and specific detection. The method developed showed unrivalled specificity with the 79 tested strains and, in contrast to previously established techniques, distinguished between phylogenetically close subspecies such as X. arboricola pv. corylina. The sensitivity of this test is comparable to that of a previously reported TaqMan^? assay at 10³ CFU mL^?1, while the unrivalled speed of LAMP technology enables a positive result to be obtained in <15 min. The developed assay can be used with real‐time fluorescent detectors for quantitative results as well as with DNA‐staining dyes to function as a simplified strategy for on‐site pathogen detection.     

A New Micromonospora Strain with Antibiotic Activity Isolated from the Microbiome of a Mid-Atlantic Deep-Sea Sponge

To tackle the growing problem of antibiotic resistance, it is essential to identify new bioactive compounds that are effective against resistant microbes and safe to use. Natural products and their derivatives are, and will continue to be, an important source of these molecules. Sea sponges harbour a diverse microbiome that co-exists with the sponge, and these bacterial communities produce a rich array of bioactive metabolites for protection and resource competition. For these reasons, the sponge microbiota constitutes a potential source of clinically relevant natural products. To date, efforts in bioprospecting for these compounds have focused predominantly on sponge specimens isolated from shallow water, with much still to be learned about samples from the deep sea. Here we report the isolation of a new Micromonospora strain, designated 28ISP2-46^T, recovered from the microbiome of a mid-Atlantic deep-sea sponge. Whole-genome sequencing reveals the capacity of this bacterium to produce a diverse array of natural products, including kosinostatin and isoquinocycline B, which exhibit both antibiotic and antitumour properties. Both compounds were isolated from 28ISP2-46^T fermentation broths and were found to be effective against a plethora of multidrug-resistant clinical isolates. This study suggests that the marine production of isoquinocyclines may be more widespread than previously supposed and demonstrates the value of targeting the deep-sea sponge microbiome as a source of novel microbial life with exploitable biosynthetic potential.     

Virus-specific cellular blastogenesis and interleukin-2 production in swine after recovery from African swine fever

Animals recovered from viral diseases represent an important model to study the host cellular and humoral immune responses to the etiologic agents. This is particularly important for African swine fever virus (ASFV) infections in which antibodies have little or no virus-neutralizing effect. Pigs surviving experimental infection with the naturally occurring low-virulent, nonhemadsorbing ASFV/NH/P68 (NHV) isolate did, however, exhibit virus-specific T-cell activities, as measured by a variety of assays. A strong virus-induced, antigen-specific blastogenic response was observed only with blood mononuclear cells (BMC) from ASF-recovered swine, whereas cells from recovered and naive swine responded similarly to the mitogens concanavalin A and phytohemagglutinin. The ASFV-induced blastogenesis was dependent on virus dose and on the presence of adherent cells. Blood mononuclear cells cultured with antigenically related hemadsorbing ASFV isolates of different virulence characteristics, the highly virulent L60 isolate and moderately virulent DRII isolate, exhibited a similar magnitude of blastogenesis to cells infected with the low-virulent NHV isolate. Virus-infected cells proved to be an efficient inducer of interleukin-2 (IL-2) activity to cells from recovered swine, but not from naive swine, whereas T-cell-specific lectins induced production of similar amounts of IL-2 activity from cells of naive and recovered swine. Correlated with the appearance of virus-induced IL-2 activity in the culture supernatant was the induction of promiscuous killing in cells exposed to prolonged (7 days) virus stimulation.(ABSTRACT TRUNCATED AT 250 WORDS)     

The variation in hectolitre weight of wheat grain fed with or without enzyme supplementation influences nutrient digestibility and subsequently affects performance in pigs

A 2 × 2 factorial experiment was conducted to investigate the effect of feeding a wheat‐based diet of two different hectolitre weights (66 vs. 74 kg/hl), achieved through different agronomical conditions, with or without the supplementation of a β‐glucanase and β‐xylanase enzyme mix on young pigs. The parameter categories which were assessed included growth performance, coefficient of apparent total tract digestibility (CATTD), faecal consistency, faecal microbial populations and faecal volatile fatty acid (VFA) concentrations. Sixty‐four pigs (11.6 kg SD 0.97) were assigned to one of four dietary treatments: (T1) low hectolitre weight wheat diet, (T2) low hectolitre weight wheat diet containing 0.1 g/kg β‐glucanase and β‐xylanase enzyme supplement, (T3) high hectolitre weight wheat diet and (T4) high hectolitre weight wheat diet containing 0.1 g/kg β‐glucanase and β‐xylanase enzyme supplement. The inclusion of wheat was 500 g/kg in the diet. The low hectolitre weight grain had a higher level of zearalenone, aflatoxin and ochratoxin contamination compared to the high hectolitre weight grain. The high hectolitre weight wheat had a higher gross energy (GE), crude protein (CP) and lysine contents compared to the low hectolitre weight wheat. Pigs offered the low hectolitre weight diet had a lower average daily gain (ADG) (p < 0.001), a lower gain to feed (G:F) ratio (p < 0.001) and a higher faecal score (more diarrhoea) (p < 0.001) compared to pigs offered the high hectolitre weight. The low hectolitre weight diet had a reduced CATTD (p < 0.05) of nitrogen (N) and gross energy (GE) compared with pigs offered the high hectolitre weight diet. In conclusion, the higher level of mycotoxins and lower content of GE, CP and lysine in the low‐quality wheat reduced ADG and the CATTD of nutrients in pigs offered this diet. The inclusion of a β‐glucanase and β‐xylanase enzyme mix had no effect on growth performance or nutrient digestibility.