Pest risk assessment of African Leucinodes species for the European Union.

Following a request from the European Commission, the EFSA Panel on Plant Health performed a quantitative risk assessment for the EU of African Leucinodes species (Lepidoptera: Crambidae), which are fruit and shoot borers, especially of eggplant type fruit. The assessment focused on (i) potential pathways for entry, (ii) distribution of infested imports within EU, (iii) climatic conditions favouring establishment, (iv) spread and (v) impact. Options for risk reduction are discussed, but their effectiveness was not quantified. Leucinodes spp. are widely distributed across sub-Saharan Africa but are little studied and they could be much more widespread in Africa than reported. Much African literature erroneously reports them as Leucinodes orbonalis which is restricted to Asia. The import of eggplant type fruit from sub-Saharan Africa consists of special fruit types and caters mostly to niche markets in the EU. The main pathway for entry is fruit of Solanum aethiopicum and exotic varieties of eggplant (S. melongena). CLIMEX modelling was used with two possible thresholds of ecoclimatic index (EI) to assess establishment potential. Climates favouring establishment occur mostly in southern Europe, where, based on human population, 14% of the imported produce is distributed across NUTS2 regions where EI ≥ 30; or where 23% of the produce is distributed where EI ≥ 15. Over the next 5 years, an annual median estimate of ~ 8600 fruits, originating from Africa, and infested with African Leucinodes spp. are expected to enter EU NUTS2 regions where EI ≥ 15 (90% CR ~ 570-52,700); this drops to ~ 5200 (90% CR ~ 350-32,100) in NUTS2 regions where EI ≥ 30. Escape of adult moths occurs mostly from consumer waste; considering uncertainties in pathway transfer, such as adult emergence, mate finding and survival of progeny, the annual median probability of a mated female establishing a founder population in NUTS regions where EI ≥ 15 was estimated to be 0.0078 (90% CR 0.00023-0.12125). This equates to a median estimate of one founder population ~ every 128 years (90% CR approximately one every 8-4280 years). Using an EI ≥ 30, the median number of founder populations establishing in the EU annually is 0.0048 (90% CR 0.0001-0.0739), equating to a median estimate of one founder population approximately every 210 years (90% CR approximately one every 14-7020 years). Under climate change for the period 2040-2059, the percent of infested produce going to suitable areas would be increased to 33% for EI ≥ 15 and to 21% for EI ≥ 30. Accordingly, the waiting time until the next founder population would be reduced to median estimates of 89 years for EI ≥ 15 (90% CR ~ 6-2980 years) and 139 years for EI ≥ 30 (90% CR 9-4655 years). If a founder population were to establish, it is estimated to spread at a rate of 0.65-7.0 km per year after a lag phase of 5-92 years. Leucinodes spp. are estimated to reduce eggplant yield by a median value of 4.5% (90% CR 0.67%-13%) if growers take no specific action, or 0.54% (90% CR between 0.13% and 1.9%) if they do take targeted action, matching previous estimates made during a risk assessment of L. orbonalis from Asia.

Risk assessment of Retithrips syriacus for the EU.

Following a request from the European Commission, the EFSA Panel on Plant Health performed a quantitative risk assessment for the EU of Retithrips syriacus (Mayet) (Thysanoptera: Thripidae), a polyphagous thrips, regarded as a tropical/subtropical pest occurring in several countries of Africa, South America, Asia and in the EU in Cyprus. The current risk assessment focused on potential pathways for entry, the climatic conditions allowing establishment, the expected spread capacity and the impact considering a time horizon of 10 years (2023-2032). The Panel identified the import of cut roses, persimmons, table grapes, as well as plants for planting of the genera Acalypha and Terminalia from third countries and those of Persea americana (avocado) from Israel as the most relevant entry pathways to consider. Over the next 10 years, an annual median estimate of 95 (90% Certainty Range, CR, ranging from 13 to 1832) potential R. syriacus founder populations per year are expected to successfully transfer to a suitable host in the EU NUTS2 regions where the climatic conditions are predicted as suitable for establishment; this value drops to a median of 4.6 founder populations per year (90% CR: 1 every 1.9 years - 85.6 per year) after considering the actual probability of establishment of a potential founder population. The estimated number of founder population per year is mostly driven by the import of cut roses and plants for planting. If such founder populations were to establish, R. syriacus is estimated to spread at a median rate of 0.05 km/year (90% CR 0.02-2.30 km/year) after a median lag phase of 1.1 years (90% CR 0.3-3.3 years). The overall impact on yield (expressed as % of the total agricultural production) directly attributable to R. syriacus when considering: (i) the main R. syriacus hosts in the EU, (ii) the areas of the EU where establishment is possible, (iii) the current agricultural practices and (iv) the evidence of impact from the countries where the pest is established for a long time, was estimated at 0.065% as the median value of the uncertainty distribution (90% CR 0.001%-0.571%). Options for risk reduction are discussed, but the effectiveness was not quantified.

Pest categorisation of Dendrolimus punctatus.

The EFSA Panel on Plant Health performed a pest categorisation of Dendrolimus punctatus (Lepidoptera: Lasiocampidae), following a commodity risk assessment of bonsai Pinus parviflora grafted onto P. thunbergii from China, in which D. punctatus was identified as a pest of possible concern to the European Union (EU). D. punctatus, also known as the Masson pine caterpillar, is present in China, Taiwan, Vietnam, India and has recently spread to Japanese islands close to Taiwan. Larval feeding on the needles of Pinus elliottii, P. luchuensis, P. massoniana, P. merkusii and P. tabulaeformis causes important damage. D. punctatus larvae can also feed on P. armandii, P. echinata, P. latteri, P. parviflora, P. sylvestris var. mongolica, P. taeda, P. taiwanensis and P. thunbergii, but full development on these hosts is uncertain. The pest has three to five generations per year; winter is spent as larvae on branch tips, on tree trunks and in the soil. The females lay egg clusters on pine needles. Pupation occurs in cocoons attached to branches or needles. D. punctatus could enter the EU either as eggs, larvae or pupae in the foliage of plants for planting or cut branches, as larvae on wood with bark or as overwintering larvae in branches, crevices in the bark or in the litter of potted plants. However, Annex VI of 2019/2072 prohibits the introduction of D. punctatus hosts (Pinus spp.) from countries and areas where the pest occurs. There are climate zones where the pest occurs in Asia that also occur in the EU, though they are limited, which constitutes an uncertainty regarding establishment. The pest's main hosts are not grown in the EU. However, the fact that it attacks the North American Pinus echinata, P. elliottii and P. taeda in its Asian native area suggests a potential capacity to shift to pine species occurring in the EU territory. D. punctatus satisfies all the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest. Whether the Pinus commonly found in Europe could act as hosts is unknown but is fundamental, affecting the criteria of establishment and magnitude of impact.

Pest categorisation of Pyrrhoderma noxium.

Following the commodity risk assessment of bonsai plants (Pinus parviflora grafted on Pinus thunbergii) from China performed by EFSA, the EFSA Plant Health Panel performed a pest categorisation of Pyrrhoderma noxium, a clearly defined plant pathogenic basidiomycete fungus of the order Hymenochaetales and the family Hymenochaetaceae. The pathogen is considered as opportunistic and has been reported on a wide range of hosts, mainly broad-leaved and coniferous woody plants, causing root rots. In addition, the fungus was reported to live saprophytically on woody substrates and was isolated as an endophyte from a few plant species. This pest categorisation focuses on the hosts that are relevant for the EU (e.g. Citrus, Ficus, Pinus, Prunus, Pyrus, Quercus and Vitis vinifera). Pyrrhoderma noxium is present in Africa, Central and South America, Asia and Oceania. It has not been reported in the EU. Pyrrhoderma noxium is not included in Commission Implementing Regulation (EU) 2019/2072. Plants for planting (excluding seeds), bark and wood of host plants as well as soil and other growing media associated with plant debris are the main pathways for the entry of the pathogen into the EU. Host availability and climate suitability factors occurring in parts of the EU are favourable for the establishment and spread of the pathogen. The introduction and spread of the pathogen into the EU are expected to have an economic and environmental impact in parts of the territory where hosts are present. Phytosanitary measures are available to prevent the introduction and spread of the pathogen into the EU. Pyrrhoderma noxium satisfies all the criteria that are within the remit of EFSA to assess for this species to be regarded as potential Union quarantine pest.

Pest risk assessment of Leucinodes orbonalis for the European Union.

Following a request from the European Commission, the EFSA Panel on Plant Health performed a quantitative risk assessment of Leucinodes orbonalis (Lepidoptera: Crambidae), the eggplant fruit and shoot borer, for the EU. The assessment focused on potential pathways for entry, climatic conditions favouring establishment, spread and impact. Options for risk reduction are discussed but effectiveness was not quantified. L. orbonalis is a key pest of eggplant (aubergine/brinjal) in the Indian subcontinent and occurs throughout most of southern Asia with records mostly from India and Bangladesh. The main pathway of entry is fruit of solanaceous plants, primarily exotic varieties of eggplant, Solanum melongena and turkey berry, S. torvum. The trade in both commodities from Asia is small but nevertheless dwarfs the trade in other Solanum fruits from Asia (S. aethiopicum, S. anguivi, S. virginianum, S. aculeatissimum, S. undatum). Other Solanum fruits were therefore not further assessed as potential pathways. The trade in eggplant from Asia consists of special fruit types and caters mostly to niche markets in the EU, while most eggplant consumed in Europe is produced in southern European and northern African countries, where L. orbonalis does not occur. Using expert knowledge elicitation (EKE) and pathway modelling, the Panel estimated that approximately 3-670 infested fruit (90% certainty range, CR) of S. melongena or fruit bunches of S. torvum enter into regions of the EU that are suitable for L. orbonalis establishment each year. Based on CLIMEX modelling, and using two possible thresholds of ecoclimatic index (EI) to indicate uncertainty in establishment potential, climates favouring establishment occur mostly in southern Europe, where, based on human population, approximately 14% of the imported produce is distributed across NUTS2 regions where EI ≥ 30; or 23% of the produce is distributed where EI ≥ 15. Escape of adult moths occurs mostly from consumer waste. By analysing results of different scenarios for the proportion of S. melongena and S. torvum in the trade, and considering uncertainties in the climatic suitability of southern Europe, adult moth emergence in areas suitable for establishment is expected to vary between 84 individuals per year and one individual per 40 years (based on 90% CR in different scenarios). In the baseline scenario, 25% of the solanaceous fruit from Asia is S. torvum, 75% is S. melongena and EI ≥ 30 is required for establishment. After accounting for the chances of mating, host finding and establishment, the probability of a mated female establishing a founder population in the EU is less than 1 in 100,000 to about 1 event per 622 years (90% CR in baseline scenario). The waiting time until the first establishment is then 622 to more than 100,000 years (CR). If such a founder population were established, the moth is estimated to spread at a rate of 0.65-7.0 km per year after a lag phase of 5-92 years. The impact of the insect on the production of eggplant is estimated to be 0.67%-13% (CR) if growers take no specific action against the insect and 0.13%-1.9% if they do take targeted actions. Tomato (S. lycopersicum) and potato (S. tuberosum) are hosts of L. orbonalis, but the insect does not develop to maturity in tomato fruit, and it does not feed on potato tubers under field conditions; hence, damage to potato can only occur due to feeding on shoots. Tomato and potato are not preferred hosts; nevertheless, impact can occur if populations of L. orbonalis are high and preferred hosts are not available. The Panel did not assess this damage due to insufficient information.

The effect of in vitro simulated colonic pH gradients on microbial activity and metabolite production using common prebiotics as substrates.

BACKGROUND: The interplay between gut microbiota (GM) and the metabolization of dietary components leading to the production of short-chain fatty acids (SCFAs) is affected by a range of factors including colonic pH and carbohydrate source. However, there is still only limited knowledge on how the GM activity and metabolite production in the gastrointestinal tract could be influenced by pH and the pH gradient increases along the colon. RESULTS: Here we investigate the effect of pH gradients corresponding to levels typically found in the colon on GM composition and metabolite production using substrates inulin, lactose, galactooligosaccharides (GOS), and fructooligosaccharide (FOS) in an in vitro colon setup. We investigated 3 different pH regimes (low, 5.2 increasing to 6.4; medium, 5.6 increasing to 6.8 and high, 6.0 increasing to 7.2) for each fecal inoculum and found that colonic pH gradients significantly influenced in vitro simulated GM structure, but the influence of fecal donor and substrate was more pronounced. Low pH regimes strongly influenced GM with the decreased relative abundance of Bacteroides spp. and increased Bifidobacterium spp. Higher in vitro simulated colonic pH promoted the production of SCFAs in a donor- and substrate-dependent manner. The butyrate producer Butyricimonas was enriched at higher pH conditions, where also butyrate production was increased for inulin. The relative abundance of Phascolarctobacterium, Bacteroides, and Rikenellaceae also increased at higher colonic pH, which was accompanied by increased production of propionate with GOS and FOS as substrates. CONCLUSIONS: Together, our results show that colonic substrates such as dietary fibres influence GM composition and metabolite production, not only by being selectively utilized by specific microbes, but also because of their SCFA production, which in turn also influences colonic pH and overall GM composition and activity. Our work provides details about the effect of the gradients of rising pH from the proximal to distal colon on fermenting dietary substrates in vitro and highlights the importance of considering pH in GM research.

Pest categorisation of Garella musculana.

The European Commission requested the EFSA Panel on Plant Health to conduct a pest categorisation of Garella musculana (Erschov) (Lepidoptera: Nolidae), following a commodity risk assessment of Juglans regia plants for planting from Türkiye, in which G. musculana was identified as a pest of possible concern to the European Union (EU). Commonly known as the Asian walnut moth, this pest is native to Central Asia and develops on shoots, buds and fruits of Juglans species such as the English walnut, J. regia and the black walnut, J. nigra. Other reported host plants, such as Prunus dulcis and Populus spp., still require confirmation. The pest was first recorded in the EU (Bulgaria) in 2016 and was then reported in Romania in 2018 and Italy in 2021. This moth completes from one to four generations per year depending on environmental conditions (from valley to mountain forests and orchards up to an altitude of 2100 m). Eggs are laid in groups of 2-3 on young nuts or on buds of 1-year-old shoots. Neonate larvae usually enter the young nut through the peduncle. After fully exploiting one nut, the larva continues feeding in another one. Development takes 25-40 days. Larvae of the autumn generation do not enter the nuts, and so feed only in the pericarp. Larvae also often feed inside 1-year-old shoots or leaf axils. Larvae develop within the host but exit to pupate under loose bark or in deep cracks of bark. The pest overwinters at the larval or pupal stages. Plants for planting, cut branches and infested nuts provide pathways for entry. Climatic conditions and availability of host plants in southern and central EU MSs have allowed this species to establish and spread in Bulgaria, Romania and Italy. Adults can fly and the pest could spread naturally within the EU. Impact on Juglans spp. cultivated for fruit, timber and ornamental purposes is anticipated. Phytosanitary measures are available to reduce the likelihood of entry and further spread of G. musculana. This species meets the criteria that are within the remit of EFSA to assess for this species to be regarded as a potential Union quarantine pest.

Pest categorisation of Pratylenchus loosi.

Following the EFSA commodity risk assessment of Malus domestica plants imported from Türkiye into the EU, the EFSA Panel on Plant Health performed a pest categorisation of Pratylenchus loosi (Nematoda: Pratylenchidae) for the EU. Pratylenchus loosi belongs to the order Rhabditida, subfamily Pratylenchidae. This nematode is not known to be present in the EU. The species is not included in the EU Commission Implementing Regulation 2019/2072. The pest occurs primarily in tropical, subtropical and warm temperate areas. It is widely distributed in Asian countries, with tea plants (Camellia sinensis) as the main host. The pest was reported from more than 60 plant species, but reports from hosts other than C. sinensis, e.g. citrus (Citrus spp.) and banana (Musa spp.), are associated with high uncertainty due to doubtful pest identification. Morphological and molecular methods are available for the identification of the pest. Pathways of entry are host plants for planting except seeds, as well as soil attached to plants for planting, machinery or footwear. Soil import to the EU is prohibited from third countries. The climatic preferences of P. loosi are compatible with the microclimatic conditions occurring in the areas of the EU where tea is grown outside. The impact of the nematode is primarily known for Asian countries, where it is a devastating pathogen on tea plants, but there is a key uncertainty on impacts on hosts other than tea. Considering the strong pathogenicity of the pest, its establishment in tea producing areas would have negative consequences for tea producers. Therefore, the Panel concludes that P. loosi satisfies all the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest.

Pest categorisation of Malacosoma parallela.

The EFSA Panel on Plant Health performed a pest categorisation of Malacosoma parallela (Staudinger) (Lepidoptera: Lasiocampidae) for the territory of the European Union, following commodity risk assessments of Berberis thunbergii, Malus domestica, Prunus persica and P. dulcis plants for planting from Türkiye, in which M. parallela came to attention as of possible concern. M. parallela is commonly known as the mountain ring silk moth and is a polyphagous leaf-eating pest in west-central Asia, primarily feeding on deciduous trees and shrubs, and known to cause serious damage to Malus, Prunus, and Quercus species. It is found at a range of altitudes from 130 m to 3000 m although most common above 1000 m. It is a univoltine species. Eggs are laid in masses on twigs and branches in the summer and larvae hatch the following spring to feed on buds and fresh leaves. Host plants can be completely defoliated. Plants for planting and cut branches provide pathways for entry, especially if infested with egg masses. Host availability and climate suitability suggest that parts of the EU would be suitable for establishment. Adults can fly and the pest could spread naturally within the EU although adults only live for a few days. Faster and more extensive spread is therefore more likely via egg masses moved on plants for planting. The introduction of M. parallela into the EU could lead to outbreaks causing damage to deciduous trees and shrubs in forests and orchards. Phytosanitary measures are available to inhibit the entry and spread of this species. M. parallela satisfies all the criteria that are within the remit of EFSA to assess for it to be regarded as a potential Union quarantine pest.

Pest categorisation of Pestalotiopsis microspora.

Following an EFSA commodity risk assessment of bonsai plants (Pinus parviflora grafted on Pinus thunbergii) imported from China, the EFSA Plant Health Panel performed a pest categorisation of Pestalotiopsis microspora, a clearly defined plant pathogenic fungus of the family Pestalotiopsidaceae. The pathogen was reported on a wide range of monocotyledonous, dicotyledonous and gymnosperms, either cultivated or wild plant species, causing various symptoms such as leaf spot, leaf blight, scabby canker, fruit spot, pre- and post-harvest fruit rot and root rot. In addition, the fungus was reported as an endophyte on a wide range of asymptomatic plant species. This pest categorisation focuses on the hosts that are relevant for the EU and for which there is robust evidence that the pathogen was formally identified by a combination of morphology, pathogenicity and multilocus sequencing analyses. Pestalotiopsis microspora was reported in Africa, North, Central and South America, Asia and Oceania. In the EU, it was reported in the Netherlands. There is a key uncertainty on the geographical distribution of P. microspora worldwide and in the EU, because of the endophytic nature of the fungus, the lack of surveys, and because in the past, when molecular tools were not fully developed, the pathogen might have been misidentified as other Pestalotiopsis species or other members of the Pestalodiopsidaceae family based on morphology and pathogenicity tests. Pestalotiopsis microspora is not included in Commission Implementing Regulation (EU) 2019/2072. Plants for planting, fresh fruits, bark and wood of host plants as well as soil and other growing media associated with plant debris are the main pathways for the entry of the pathogen into the EU. Host availability and climate suitability in parts of the EU are favourable for the establishment and spread of the pathogen. The introduction and spread of the pathogen into the EU are expected to have an economic and environmental impact where susceptible hosts are grown. Phytosanitary measures are available to prevent the introduction and spread of the pathogen into the EU. Unless the restricted distribution in the EU is disproven, Pestalotiopsis microspora satisfies all the criteria that are within the remit of EFSA to assess for this species to be regarded as potential Union quarantine pest.

Pest categorisation of Pestalotiopsis disseminata.

Following the commodity risk assessments of bonsai plants from China consisting of Pinus parviflora grafted on P. thunbergii performed by EFSA, the EFSA Plant Health Panel performed a pest categorisation of Pestalotiopsis disseminata, a clearly defined plant pathogenic fungus of the family Pestalotiopsidaceae. The pathogen has been reported on herbaceous, woody and ornamental plants causing symptoms such as leaf blight, shoot blight, seedling blight, pod canker, pre- and post-harvest fruit rot, and gummosis. Moreover, the fungus was reported as an endophyte on a wide range of asymptomatic hosts. The pathogen is present in Africa, North and South America, Asia, Europe and Oceania. It has been reported from the EU, with a restricted distribution (Portugal). There is a key uncertainty on the geographical distribution of P. disseminata in the EU and worldwide, because of the endophytic nature of the fungus, the lack of surveys and since the pathogen might have been misidentified based only on morphology and pathogenicity tests. The pathogen is not included in Commission Implementing Regulation (EU) 2019/2072. This pest categorisation focuses on those hosts that are relevant for the EU and for which there is robust evidence that the pathogen was formally identified by a combination of morphology, pathogenicity and multilocus sequence analysis. Plants for planting, fresh fruits, bark and wood of host plants as well as soil and other plant growing media are the main pathways for the entry of the pathogen into the EU. Host availability and climate suitability factors occurring in parts of the EU are favourable for the establishment of the pathogen. Despite the low aggressiveness observed in most reported hosts, and the fact that P. disseminata may colonise plants as an endophyte, its introduction and spread in the EU may have an economic and environmental impact (with a key uncertainty) where susceptible hosts are grown. Phytosanitary measures are available to prevent the introduction and spread of the pathogen. The Panel cannot conclude on whether P. disseminata satisfies all the criteria that are within the remit of EFSA to assess for this species to be regarded as potential Union quarantine pest, because of the key uncertainties on the restricted distribution in the EU and the magnitude of the impact.

The impact of storage buffer and storage conditions on fecal samples for bacteriophage infectivity and metavirome analyses.

BACKGROUND: There is an increasing interest in investigating the human gut virome for its influence on the gut bacterial community and its putative influence on the trajectory towards health or disease. Most gut virome studies are based on sequencing of stored fecal samples. However, relatively little is known about how conventional storage buffers and storage conditions affect the infectivity of bacteriophages and influence the downstream metavirome sequencing. RESULTS: We demonstrate that the infectivity and genome recovery rate of different spiked bacteriophages (T4, c2 and Phi X174) are variable and highly dependent on storage buffers. Regardless of the storage temperature and timespan, all tested phages immediately lost 100% (DNA/RNA Shield) or more than 90% (StayRNA and RNAlater) of their infectivity. Generally, in SM buffer at 4 °C phage infectivity was preserved for up to 30 days and phage DNA integrity was maintained for up to 100 days. While in CANVAX, the most effective buffer, all spiked phage genomes were preserved for at least 100 days. Prolonged storage time (500 days) at - 80 °C impacted viral diversity differently in the different buffers. Samples stored in CANVAX or DNA/RNA Shield buffer had the least shifts in metavirome composition, after prolonged storage, but they yielded more contigs classified as "uncharacterised". Moreover, in contrast to the SM buffer, these storage buffers yielded a higher fraction of bacterial DNA in metavirome-sequencing libraries. We demonstrated that the latter was due to inactivation of the DNases employed to remove extra-cellular DNA during virome extraction. The latter could be partly avoided by employing additional washing steps prior to virome extraction. CONCLUSION: Fecal sample storage buffers and storage conditions (time and temperature) strongly influence bacteriophage infectivity and viral composition as determined by plaque assay and metavirome sequencing. The choice of buffer had a larger effect than storage temperature and storage time on the quality of the viral sequences and analyses. Based on these results, we recommend storage of fecal virome samples at in SM buffer at 4 °C for the isolation of viruses and at - 80 °C for metagenomic applications if practically feasible (i.e., access to cold storage). For fecal samples stored in other buffers, samples should be cleared of these buffers before viral extraction and sequencing. Video Abstract.

Dietary non-starch polysaccharides impair immunity to enteric nematode infection.

BACKGROUND: The influence of diet on immune function and resistance to enteric infection and disease is becoming ever more established. Highly processed, refined diets can lead to inflammation and gut microbiome dysbiosis, whilst health-promoting dietary components such as phytonutrients and fermentable fibres are thought to promote a healthy microbiome and balanced mucosal immunity. Chicory (Cichorium intybus) is a leafy green vegetable rich in fibres and bioactive compounds that may promote gut health. RESULTS: Unexpectedly, we here show that incorporation of chicory into semisynthetic AIN93G diets renders mice susceptible to infection with enteric helminths. Mice fed a high level of chicory leaves (10% dry matter) had a more diverse gut microbiota, but a diminished type-2 immune response to infection with the intestinal roundworm Heligmosomoides polygyrus. Furthermore, the chicory-supplemented diet significantly increased burdens of the caecum-dwelling whipworm Trichuris muris, concomitant with a highly skewed type-1 immune environment in caecal tissue. The chicory-supplemented diet was rich in non-starch polysaccharides, particularly uronic acids (the monomeric constituents of pectin). In accordance, mice fed pectin-supplemented AIN93G diets had higher T. muris burdens and reduced IgE production and expression of genes involved in type-2 immunity. Importantly, treatment of pectin-fed mice with exogenous IL-25 restored type-2 responses and was sufficient to allow T. muris expulsion. CONCLUSIONS: Collectively, our data suggest that increasing levels of fermentable, non-starch polysaccharides in refined diets compromises immunity to helminth infection in mice. This diet-infection interaction may inform new strategies for manipulating the gut environment to promote resistance to enteric parasites.

Pruning Wound Protection Products Induce Alterations in the Wood Mycobiome Profile of Grapevines.

Fungal pathogens involved in grapevine trunk diseases (GTDs) may infect grapevines throughout their lifetime, from nursery to vineyard, via open wounds in stems, canes or roots. In vineyards, pruning wound protection products (PWPPs) offer the best means to reduce the chance of infection by GTD fungi. However, PWPPs may affect non-target microorganisms that comprise the natural endophytic mycobiome residing in treated canes, disrupting microbial homeostasis and indirectly influencing grapevine health. Using DNA metabarcoding, we characterized the endophytic mycobiome of one-year-old canes of cultivars Cabernet Sauvignon and Syrah in two vineyards in Portugal and Italy and assessed the impact of established and novel PWPPs on the fungal communities of treated canes. Our results reveal a large fungal diversity (176 taxa), and we report multiple genera never detected before in grapevine wood (e.g., Symmetrospora and Akenomyces). We found differences in mycobiome beta diversity when comparing vineyards (p = 0.01) but not cultivars (p > 0.05). When examining PWPP-treated canes, we detected cultivar- and vineyard-dependent alterations in both alpha and beta diversity. In addition, numerous fungal taxa were over- or under-represented when compared to control canes. Among them, Epicoccum sp., a beneficial genus with biological control potential, was negatively affected by selected PWPPs. This study demonstrates that PWPPs induce alterations in the fungal communities of grapevines, requiring an urgent evaluation of their direct and indirect effects on plants health with consideration of factors such as climatic conditions and yearly variations, in order to better advise viticulturists and policy makers.

A global microbiome survey of vineyard soils highlights the microbial dimension of viticultural terroirs.

The microbial biodiversity found in different vitivinicultural regions is an important determinant of wine terroir. It should be studied and preserved, although it may, in the future, be subjected to manipulation by precision agriculture and oenology. Here, we conducted a global survey of vineyards' soil microbial communities. We analysed soil samples from 200 vineyards on four continents to establish the basis for the development of a vineyard soil microbiome's map, representing microbial biogeographical patterns on a global scale. This study describes vineyard microbial communities worldwide and establishes links between vineyard locations and microbial biodiversity on different scales: between continents, countries, and between different regions within the same country. Climate data correlates with fungal alpha diversity but not with prokaryotes alpha diversity, while spatial distance, on a global and national scale, is the main variable explaining beta-diversity in fungal and prokaryotes communities. Proteobacteria, Actinobacteria and Acidobacteria phyla, and Archaea genus Nitrososphaera dominate prokaryotic communities in soil samples while the overall fungal community is dominated by the genera Solicoccozyma, Mortierella and Alternaria. Finally, we used microbiome data to develop a predictive model based on random forest analyses to discriminate between microbial patterns and to predict the geographical source of the samples with reasonable precision.

Combining reverse osmosis and microbial degradation for remediation of drinking water contaminated with recalcitrant pesticide residue.

Groundwater contamination by recalcitrant organic micropollutants such as pesticide residues poses a great threat to the quality of drinking water. One way to remediate drinking water containing micropollutants is to bioaugment with specific pollutant degrading bacteria. Previous attempts to augment sand filters with the 2,6-dichlorobenzamide (BAM) degrading bacterium Aminobacter niigataensis MSH1 to remediate BAM-polluted drinking water initially worked well, but the efficiency rapidly decreased due to loss of degrader bacteria. Here, we use pilot-scale augmented sand filters to treat retentate of reverse osmosis treatment, thus increasing residence time in the biofilters and potentially nutrient availability. In a first pilot-scale experiment, BAM and most of the measured nutrients were concentrated 5-10 times in the retentate. This did not adversely affect the abundances of inoculated bacteria and the general prokaryotic community of the sand filter presented only minor differences. On the other hand, the high degradation activity was not prolonged compared to the filter receiving non-concentrated water at the same residence time. Using laboratory columns, it was shown that efficient BAM degradation could be achieved for >100 days by increasing the residence time in the sand filter. A slower flow may have practical implications for the treatment of large volumes of water, however this can be circumvented when treating only the retentate water equalling 10-15% of the volume of inlet water. We therefore conducted a second pilot-scale experiment with two inoculated sand filters receiving membrane retentate operated with different residence times (22 versus 133 min) for 65 days. While the number of MSH1 in the biofilters was not affected, the effect on degradation was significant. In the filter with short residence time, BAM degradation decreased from 86% to a stable level of 10-30% degradation within the first two weeks. The filter with the long residence time initially showed >97% BAM degradation, which only slightly decreased with time (88% at day 65). Our study demonstrates the advantage of combining membrane filtration with bioaugmented filters in cases where flow rate is of high importance.

Gluten-free diet reduces autoimmune diabetes mellitus in mice across multiple generations in a microbiota-independent manner.

Experimental and clinical data suggest that a gluten-free diet attenuates the development of type 1 diabetes. A gluten-free diet changes the gut microbiota composition, and such microbial changes are expected to reduce the autoimmune responses. However, in experiments with laboratory mice, a gluten-free diet changes the gut microbiota differently under varying experimental settings, questioning the specific role of the gut microbes. Here we show that a maternal gluten-free diet until weaning of their pups, delayed type 1 diabetes in both dams (parent generation) and offspring (F1 generation) of untreated non-obese diabetic (NOD) mice and in mice treated with a full cocktail of antibiotics that eradicates most of the existing microbiota. Breeding a second (F2) generation of NOD mice, never exposed to the gluten-free diet or the associated microbial changes, also demonstrated a preventative effect on type 1 diabetes even though their parents (the F1 generation) had only been on a gluten-free diet very early in life. Collectively, the experimental data, thus, points towards microbiota-independent dietary protection. Furthermore, both the perinatal gluten-free diet and antibiotic treatment reduced inflammation in the salivary glands and improved glucose challenged beta cell function in the F1 offspring. However, in contrast to the autoimmune response in the pancreas, those changes appeared to be microbiota dependent, as they were missing in the antibiotic treated mice, and do, therefore, not seem to be related to the preventative effect on type 1 diabetes. Interestingly, adoptive transfer of splenocytes from gluten-free fed mice protected NOD.SCID mice from developing diabetes, demonstrating that the anti-diabetic effect of a gluten-free diet was based on early life changes in the evolving immune system. In particular, genes involved in regulation of lymphocyte activation, proliferation, and cell adhesion were highly expressed in the spleen in gluten-free fed mice at weaning compared to control fed mice of the F1 generation, which suggested that gluten promotes autoimmunity by inhibiting immune regulation, though the involvement of the specific genes needs further investigation. In conclusion, gluten-free diet reduces autoimmune inflammation in salivary glands and pancreas in NOD mice in a microbiota-dependent and -independent manner respectively, and has preventative effect on type 1 diabetes by modulating the systemic immune system.

Bacteriophages Roam the Wheat Phyllosphere.

The phyllosphere microbiome plays an important role in plant fitness. Recently, bacteriophages have been shown to play a role in shaping the bacterial community composition of the phyllosphere. However, no studies on the diversity and abundance of phyllosphere bacteriophage communities have been carried out until now. In this study, we extracted, sequenced, and characterized the dsDNA and ssDNA viral community from a phyllosphere for the first time. We sampled leaves from winter wheat (Triticum aestivum), where we identified a total of 876 virus operational taxonomic units (vOTUs), mostly predicted to be bacteriophages with a lytic lifestyle. Remarkably, 848 of these vOTUs corresponded to new viral species, and we estimated a minimum of 2.0 × 106 viral particles per leaf. These results suggest that the wheat phyllosphere harbors a large and active community of novel bacterial viruses. Phylloviruses have potential applications as biocontrol agents against phytopathogenic bacteria or as microbiome modulators to increase plant growth-promoting bacteria.

Influence of vintage, geographic location and cultivar on the structure of microbial communities associated with the grapevine rhizosphere in vineyards of San Juan Province, Argentina.

Soil microbiomes, as a primary reservoir for plant colonizing fungi and bacteria, play a major role in determining plant productivity and preventing invasion by pathogenic microorganisms. The use of 16S rRNA and ITS high-throughput amplicon sequencing for analysis of complex microbial communities have increased dramatically in recent years, establishing links between wine specificity and, environmental and viticultural factors, which are framed into the elusive terroir concept. Given the diverse and complex role these factors play on microbial soil structuring of agricultural crops, the main aim of this study is to evaluate how external factors, such as vintage, vineyard location, cultivar and soil characteristics, may affect the diversity of the microbial communities present. Additionally, we aim to compare the influence these factors have on the structuring of bacterial and fungal populations associated with Malbec grapevine rhizosphere with that of the more widespread Cabernet Sauvignon grapevine cultivar. Samples were taken from Malbec and Cabernet Sauvignon cultivars from two different vineyards in the San Juan Province of Argentina. Total DNA extracts from the rhizosphere soil samples were sequenced using Illumina's Miseq technology, targeting the V3-V4 hypervariable 16S rRNA region in prokaryotes and the ITS1 region in yeasts. The major bacterial taxa identified were Proteobacteria, Bacteroidetes and Firmicutes, while the major fungal taxa were Ascomycetes, Basidiomycetes, Mortierellomycetes and a low percentage of Glomeromycetes. Significant differences in microbial community composition were found between vintages and vineyard locations, whose soils showed variances in pH, organic matter, and content of carbon, nitrogen, and absorbable phosphorus.

Under the Christmas Tree: Belowground Bacterial Associations With Abies nordmanniana Across Production Systems and Plant Development.

Abies nordmanniana is an economically important tree crop widely used for Christmas tree production. After initial growth in nurseries, seedlings are transplanted to the field. Rhizosphere bacterial communities generally impact the growth and health of the host plant. However, the dynamics of these communities during A. nordmanniana growth in nurseries, and during transplanting, has not previously been addressed. By a 16S rRNA gene amplicon sequencing approach, we characterized the composition and dynamics of bacterial communities in the rhizosphere during early plant growth in field and greenhouse nurseries and for plants transplanted from the greenhouse to the field. Moreover, the N-cycling potential of rhizosphere bacteria across plant age was addressed in both nurseries. Overall, a rhizosphere core microbiome of A. nordmanniana, comprising 19.9% of the taxa at genus level, was maintained across plant age, nursery production systems, and even during the transplantation of plants from the greenhouse to the field. The core microbiome included the bacterial genera Bradyrhizobium, Burkholderia, Flavobacterium, Pseudomonas, Rhizobium, Rhodanobacter, and Sphingomonas, which harbor several N-fixing and plant growth-promoting taxa. Nevertheless, both plant age and production system caused significant changes in the rhizosphere bacterial communities. Concerning community composition, the relative abundance of Rhizobiales (genera Rhizobium, Bradyrhizobium, and Devosia) was higher in the rhizosphere of field-grown A. nordmanniana, whereas the relative abundance of Enterobacteriales and Pseudomonadales (genus Pseudomonas) was higher in the greenhouse. Analysis of community dynamics across plant age showed that in the field nursery, the most abundant bacterial orders showed more dynamic changes in their relative abundance in the rhizosphere than in the bulk soil. In the greenhouse, age-dependent dynamics even occurred but affected different taxa than for the field-grown plants. The N-cycling potential of rhizosphere bacterial communities showed an increase of the relative abundance of genes involved in nitrogen fixation and denitrification by plant age. Similarly, the relative abundance of reported nitrogen-fixing or denitrifying bacteria increased by plant age. However, different community structures seemed to lead to an increased potential for nitrogen fixation and denitrification in the field versus greenhouse nurseries.