Does the provision of live black soldier fly and yellow mealworm larvae improve Muscovy duck welfare?

Background The provision of environmental enrichments to Muscovy ducks could reduce the expression of the aggressive behaviors.The aim of the present study was to evaluate the effects of black soldier fly(BSF)and yellow mealworm(YM)live larva provision on Muscovy duck performance,excreta corticosterone metabolites(ECM),behavior,and blood parameters.Methods A total of 1263-day-old female Muscovy ducklings were allotted to 18 pens(6 replicates/treatment,7 birds/pen)and assigned to 3 experimental treatments:a control group fed commercial feed,and two experimental treatments fed commercial feed plus the 5%(based on the expected daily feed intake,as fed basis)of BSF and YM live larvae(BSF and YM groups,respectively).A two-phase feeding program was applied:starter(from 3 to 31 days of age)and grower-finisher(from 32 to 55 days of age).The live weight,average daily gain,average daily feed intake,and feed conversion ratio were calculated.Larva consumption times were collected,and video recordings were performed during 3 periods(P)each day:the hour before(P1),during(P2),and after(P3)the larva administration.ECM were evaluated at 3,31,and 55-day-old.Finally,the total red and white blood cell counts,serum proteins,lipids,and liver and renal function serum enzymes were evaluated on 12 birds/treatment.Results The experimental treatment did not affect the growth performance of the birds(P>0.05).Larva consumption times were always similar between the two insect species,except at 14–18 days of age,were BSF larvae were consumed faster than YM larvae(P<0.001).The birds showed less walking activity during P2,and preening behavior increased in YM birds during P3.The C birds increased the attack behavior over the weeks(P<0.05).During weeks 1–3 the YM group reduced the attack frequency(P1>P3;P<0.05).Finally,the provision of live BSF and YM larvae significantly reduced the ECM at 55 days of age and the heterophil to lymphocyte ratio(P<0.05).Conclusions Live BSF and YM larva supplementation in Muscovy duck improves duck welfare,without impairing birds'growth performance.

Nutritional value of a partially defatted and a highly defatted black soldier fly larvae (Hermetia illucens L.) meal for broiler chickens: apparent nutrient digestibility,apparent metabolizable energy and apparent ileal amino acid digestibility

Background: The study aimed to determine the apparent total tract digestibility coefficients(ATTDC) of nutrients, the apparent metabolizable energy(AME and AMEn) and the amino acid(AA) apparent ileal digestibility coefficients(AIDC)of a partially defatted(BSFp) and a highly defatted(BSFh) black soldier fly larvae meal. The experimental diets were: a basal diet and two diets prepared by substituting 250 g/kg(w/w) of the basal diet with BSFp or BSFh, respectively.Results: Significant differences were found between BSFp and BSFh meals for ATTDC of the nutrients: BSFp resulted more digestible than BSFh, except for ATTDC of CP which did not differed between meals, while a statistical trend was observed for ATTDC of DM and EE. The AME and AMEn values were significantly(P < 0.05) different between the two BSF meals, with higher levels for BSFp(16.25 and 14.87 MJ/kg DM, respectively). The AIDC of the AA in BSFp ranged from 0.44 to 0.92, while in BSFh they ranged from 0.45 to 0.99. No significant differences were observed for the AA digestibility(0.77 and 0.80 for BSFp and BSFh, respectively), except for glutamic acid, proline and serine that were more digestible in the BSFh meal(P < 0.05).Conclusions: Defatted BSF meals can be considered as an excellent source of AME and digestible AA for broilers with a better efficient nutrient digestion. These considerations suggested the effective utilization of defatted BSF larvae meal in poultry feed formulation.

Black soldier fly defatted meal as a dietary protein source for broiler chickens:Effects on growth performance,blood traits,gut morphology and histological features

Background: The present study has evaluated the effects of different inclusion levels of a partially defatted black soldier fly(Hermetia illucens L.; HI) larva meal on the growth performance, blood parameters and gut morphology of broiler chickens. A total of 256 male broiler chickens(Ross 308) were reared from d 1 to d 35 and assigned to4 dietary treatments(8 replicates/treatment and 8 birds/replicate). HI larva meal was included at increasing levels(0, 5%, 10% and 15%; HI0, HI5, HI10 and HI15, respectively) in isonitrogenous and isoenergetic diets formulated for 3 feeding phases: starter(1–10 d), growing(10–24 d) and finisher(24–35 d). Two birds per pen were slaughtered at d 35 and morphometric investigations and histopathological alterations were performed.Results: The live weight(LW) showed linear and quadratic responses to increasing HI larva meal(maximum for HI10 group). Average daily gain(ADG) showed a linear and quadratic responses to HI meal(maximum for HI10 group) during starter and growing periods. A linear decrease was observed for ADG during the finisher period. The daily feed intake(DFI) showed a linear and quadratic effect during the starter period(maximum for HI10 group).Linear and quadratic responses were observed for the feed conversion ratio(FCR) in the growing period and for the whole period of the experiment. The FCR showed a linear response in the finisher period(maximum for HI15).No significant effects were observed for the blood and serum parameters, except for the phosphorus concentration,which showed linear and quadratic responses as well as glutathione peroxidase(GPx) activity, the latter of which showed a linear response. The HI15 birds showed a lower villus height, a higher crypt depth and a lower villus height-to-crypt depth ratio than the other groups.Conclusions: Increasing levels of dietary HI meal inclusion in male broiler chickens may improve the LW and DFI during the starter period, but may also negatively affect the FCR and gut morphology, thus suggesting that low levels may be more suitable. However, no significant effects on the haematochemical parameters or histological findings were observed in relation to HI meal utilization.

Nutritional effects of the dietary inclusion of partially defatted Hermetia illucens larva meal in Muscovy duck

Background: The present work is aimed at evaluating the effect of different inclusion levels of a partially defatted black soldier fly(Hermetia illucens,L.;HI) larva meal for ducks.A total of 192 female 3-day-old Muscovy ducklings(Cairina moschata domestica,Canedins R71 L White,Grimaud Freres Selection,France) were divided into 4 groups,assigned 4 different dietary treatments(6 replicates/treatment and 8 birds/replicate) and reared from 3 to 50 days of age.HI larva meal was included at increasing levels(0,3%,6% and 9%,HI0,HI3,HI6 and HI9,respectively) in isonitrogenous and isoenergetic diets formulated for 3 feeding phases: starter(3–17 days of age),grower(18–38 days of age) and finisher(39–50 days of age).The growth performance and apparent total tract digestibility were evaluated during the trial using titanium dioxide as an inert marker(0.3% of inclusion).At 51 days of age,two birds per pen were slaughtered and histomorphological investigations were performed.Results: The live weight and average daily gain showed a quadratic response to increasing HI meal in the grower period(minimum corresponding to the HI6 group).No effects of dietary inclusion levels were observed for the daily feed intake or feed conversion ratio.The apparent dry matter and organic matter digestibility were not affected by the dietary treatment.A linear decrease was observed for the crude protein apparent digestibility in the starter period(minimum for the HI9 groups).The ether extract apparent digestibility increased linearly during the grower and finisher periods(minimum for the HI0 group).The morphometric indices were not influenced by the dietary treatments.Conclusions: The inclusion of up to 9% of HI partially defatted larva meal in the diet of ducks did not cause any effect on growth performance,as well as the apparent digestibility.Moreover,dietary HI inclusion preserved the physiological intestinal development.

Black soldier fly and gut health in broiler chickens:insights into the relationship between cecal microbiota and intestinal mucin composition

Background:The relationship between diet and intestinal microbiota and mucin composition appears to be fundamental for poultry gut health.The effects of insect meal(whose role as alternative feed ingredient is now well recognized)on gut microbiota and mucin composition have recently been reported in Tenebrio molitor-fed free-range and broiler chickens,but no data are currently available for Hermetia illucens(HI)-fed broilers.The present study evaluated the effects of dietary HI meal inclusion on cecal microbiota and intestinal mucin composition of broiler chickens.Results:A total of 256 male broiler chickens were allotted to 4 dietary treatments(control diet[C]and 5%,10%and 15%HI meal inclusion,with 8 replicate pens/treatment and 8 birds/pen)and slaughtered at 35 d of age(2 animals/pen,16 birds/diet).The cecal microbiota assessment by 16S rRNA amplicon based sequencing showed lower alpha diversity in HI15 chickens(Shannon,P<0.05)and higher beta diversity(Adonis and ANOSIM,P<0.001)in birds fed HI diets than C.Furthermore,HI15 birds displayed significant increase of the relative abundance of Proteobacteria phylum(False Discovery Rate[FDR]<0.05)when compared to HI10.L-Ruminococcus(Ruminococcus from Lachnospiraceae family),Faecalibacterium,Blautia and Clostridium genera were found to be characteristic of HI5 cecal microbiota(FDR<0.05),while broiler chickens fed HI10 and HI15 diets were characterized(FDR<0.05)by Lactobacillus and Ruminococcus(HI10)and Bacteroides,Roseburia and Helicobacter genera(HI15).Periodic-acid Schiff,Alcian Blue pH 2.5 and high iron diamine staining on small and large intestine also demonstrated lower mucin staining intensity in the intestinal villi of HI10 and HI15 birds than C(P<0.05).Conclusions:Dietary HI meal utilization at low inclusion levels(i.e.,5%)positively influenced either the cecal microbiota or the gut mucin dynamics in terms of selection of potentially beneficial bacteria and increase in villi mucins.However,high inclusion levels(in particular the 15%)may have a negative influence in terms of partial reduction of microbial complexity,reduction of potentially beneficial bacteria,selection of bacteria with mucolytic activity and decrease in villi mucins.

Effects of dietary Hermetia illucens meal inclusion on cecal microbiota and small intestinal mucin dynamics and infiltration with immune cells of weaned piglets

Background: The constant interaction between diet and intestinal barrier has a crucial role in determining gut health in pigs. Hermetia illucens(HI) meal(that represents a promising, alternative feed ingredient for production animals) has recently been demonstrated to influence colonic microbiota, bacterial metabolite profile and mucosal immune status of pigs, but no data about modulation of gut mucin dynamics are currently available. The present study evaluated the effects of dietary HI meal inclusion on the small intestinal mucin composition of piglets, as well as providing insights into the cecal microbiota and the mucosal infiltration with immune cells.Results: A total of 48 weaned piglets were randomly allotted to 3 dietary treatments(control diet [C] and 5% or10% HI meal inclusion [HI5 and HI10], with 4 replicate boxes/treatment and 4 animals/box) and slaughtered after61 days of trial(3 animals/box, 12 piglets/diet). The cecal microbiota assessment by 16 S r RNA amplicon based sequencing showed higher beta diversity in the piglets fed the HI-based diets than the C(P < 0.001). Furthermore,the HI-fed animals showed increased abundance of Blautia, Chlamydia, Coprococcus, Eubacterium, Prevotella,Roseburia, unclassified members of Ruminococcaceae, Ruminococcus and Staphylococcus when compared to the C group(FDR < 0.05). The gut of the piglets fed the HI-based diets showed greater neutral mucin percentage than the C(P < 0.05), with the intestinal neutral mucins of the HI-fed animals being also higher than the sialomucins and the sulfomucins found in the gut of the C group(P < 0.05). Furthermore, the piglets fed the HI-based diets displayed lower histological scores in the jejunum than the other gut segments(ileum [HI5] or ileum and duodenum [HI10], P < 0.05).Conclusions: Dietary HI meal utilization positively influenced the cecal microbiota and the small intestinal mucin dynamics of the piglets in terms of selection of potentially beneficial bacteria and preservation of mature mucin secretory architecture, without determining the development of gut inflammation. These findings further confirm the suitability of including insect meal in swine diets.

Effect of dietary supplementation with insect fats on growth performance,digestive efficiency and health of rabbits

Background: The present work aimed at evaluating the effect of the dietary replacement of soybean oil(S) by two types of insect fats extracted from black soldier fly larvae(H, Hermetia il ucens L.) and yel ow mealworm larvae(T, Tenebrio molitor L.) on growth performance, nutrient digestibility, blood parameters, intestinal morphology and health of growing rabbits.Methods: At weaning, 200 crossbred rabbits(36 days old) were allotted to five dietary treatments(40 rabbits/group): a control diet(C) containing 1.5% of soybean oil and four experimental diets where soybean oil was partially(50%) or totally(100%) substituted by H(H50 and H100) or T(T50 and T100) fats. Total tract digestibility was evaluated on 12 rabbits per treatment. The growth trial lasted 41 d and, at slaughtering(78 days old), blood samples were collected from 15 rabbits per treatment, morphometric analyses were performed on duodenum, jejunum and ileum mucosa,and samples of liver, spleen and kidney were submitted to histological evaluation.Results: No difference was observed between the control and the experimental groups fed insect fats in terms of performance, morbidity, mortality and blood variables. The addition of H and T fats did not influence apparent digestibility coefficients of dry matter, protein, ether extract, fibre fractions and gross energy. Gut morphometric indices and organ histopathology were not affected by dietary inclusion of H and T fats.Conclusions: H and T fats are suitable sources of lipid in rabbit diets to replace soybean oil without any detrimental effect on growth performance, apparent digestibility, gut mucosa traits and health.

Alternative stable states in mountain forest ecosystems： the case of European larch(Larix decidua) forests in the western Alps

European larch(Larix decidua) forests of the western Alps form extensivecultural landscapes whose resilience to global changes is currently unknown. Resilience describes the capacity of ecological systems to maintain the same state, i.e., the same function, processes, structure, and composition despite disturbances, environmental changes and internal fluctuations. Our aim is to explore the resilience of larch forests to changes in climate and land use in the western Italian Alps.To do so, we examined whether larch forests can be described as an alternative stable state in mountain forest ecosystems. We used tree basal area data obtained from field forest inventories in combination with topography, forest structure, land use, and climate information. We applied three different probabilistic methods: frequency distributions, logistic regressions, and potential analyses to infer the resilience of larch forests relative to that of other forest types.We found patters indicative of alternative stable states: bimodality in the frequency distribution of the percent of larch basal area, and the presence of an unstable state, i.e., mixed larch forests, in the potential analyses. We also found:(1) high frequency ofpurelarchforestsathighelevation,(2)the probability of pure larch forests increased mostly with elevation, and(3) pure larch forests were a stable state in the upper montane and subalpine belts.Our study shows that the resilience of larch forests may increase with elevation, most likely due to the altitudinal effect on climate. Under the same climate conditions, land use seems to be the main factor governing the dominance of larch forests. In fact, subalpine larch forests may be more resilient, and natural succession after land abandonment, e.g., towards Pinuscembra forests, seems slower than in montane larch forests. In contrast, in the upper montane belt only intense land use regimes characterized by open canopies and forest grazing may maintain larch forests.We conclude that similar approaches could be applied in other forest ecosystems to infer the resilience of tree species.

Dried fruit pomace inclusion in poultry diet:growth performance, intestinal morphology and physiology

Background: Fruit pomaces are by-products rich in polyphenol compounds and dietary fiber. They seem to play an important role in regulating the gut microbiota, morphology and physiology. The aim of this study was to assess whether apple(A), blackurrant(B) or strawberry(S) pomaces could be suitable ingredients in broiler diets and their effect on gut health. A total of 480 male broilers were randomly allotted to 8 dietary treatments with lower(3%-L) or higher(6%-H) dietary fiber content: two control groups(CL/CH), two A diets(AL/AH), two B diets(BL/BH), two S diets(SL/SH). Diet and fruit pomaces were chemically analyzed to assess polyphenol concentration and fibre fraction content. After the evaluation of growth performance, 6 birds/group were slaughtered at 35 days of age. Morphometric and histopathological investigations were performed on duodenum, jejunum and ileum.Excreta were collected to perform microbiota evaluation by 16 S DNA sequencing. Weight, viscosity, enzymatic activity, short chain fatty acid(SCFAs) and ammonia concentration were determined in ileum and/or ceca content.Results: A pomace and A diets showed the lowest polyphenol content and the highest content of soluble fibre fraction. No significant differences were observed for growth performance, gut morphometry and histopathology(P > 0.05). Dietary fruit pomace inclusion increased the weight of ileum and ceca and the ileum digesta viscosity(P < 0.05). In the ileum, A and S groups showed lower bacterial α-glucosidase activity than C groups. Moreover,small intestine SCFAs concentration was higher in fruit pomaces diets(P < 0.05). In ceca, B and S groups showed lower ammonia concentration and higher SCFAs than C. Dietary treatments also influenced the activity of α-glucosidase, α-galactosidase, β-galactosidase β-glucuronidase and xylase. Regarding microbiota, at phylum level,Firmicutes were differentially abundant across treatment(maximum for C and minimum in S, FDR > 0.05). At genus level, an increase of Weissella in AH and Erwinia in S/B diets, as well as a decrease of Lactobacillus in all fruit pomace groups were recorded(P < 0.05).Conclusions: Fruit pomaces could be suitable ingredients in poultry nutrition even if further studies are needed to better understand which doses is more recommended to avoid negative effects on gut microbiota.

Characterization of organic-rich mineral debris revealed by rapid glacier retreat, Indren Glacier, European Alps

In the summer of 2003 and 2004, characterized by a rapid glacier retreat, a stony surface covered by well-structured organic-rich mineral debris was observed very close to the Indren glacier terminus(Monte Rosa Massif, NW Italy, 3100 m ASL), on an area covered by the glacier tongue till the year before. The origin and type of this organicrich material were investigated, in order to detect their characteristics, potential sources and fate within the foreland system. The deposits were dated using Carbon-14 and analyzed for the chemical characteristics of the organic component, the elemental composition of the mineral fraction and presence of microbial markers. The material, granular and dark in color, had a total organic carbon(TOC) content ranging between 17.4 ± 0.39 and 28.1 ± 0.63 g kg^(-1) dry weight(dw), significantly higher than the surrounding glacial till(~ 1.4 g kg^(-1) dw), although only 0.33% of it was in water soluble form. Microbial carbon(C) and nitrogen(N) accounted for 10.6% and 3.13% of TOC and total N, respectively. Dissolved nitrogen(N), mainly present as ammonium, represented 2.40% of the total N. The low aromatic component and large presence of nitrogen(N)-derived compounds suggested that most of the organic carbon(OC) in these organic-rich mineral deposits was derived from microbial cells, although the high average radiocarbon age of about 2900 years may also point to the contribution of aeolian depositions of anthropogenic or natural origin. Elemental composition and the crustal enrichment factor of trace elements in the mineral fraction of the aggregates corroborated the hypothesis that most part of the accumulated material derived from ice meltwater. Some indicators of the colonization of these deposits by microbial communities were also reported, from the abundance of DNA and phylogenetic markers, to the presence of bacterial taxa commonly able to thrive in similar habitats. All these elements suggested that such kind of deposits may have a potential role as energy and nutrient sources in recently deglaciated areas, highlighting the necessity to better understand the processes underlying their formation and their evolution.

Narcea—an unknown,ancient cultivated rose variety from northern Spain

The present work reports the discovery and the complete characterisation of an ancient cultivated rose variety found growing in a private garden in the southwest of the Principality of Asturias(northern Spain).The variety is here given the name Narcea.The majority of roses currently cultivated belong to the so-called group of‘Modern Roses’,all of which were obtained after 1867 via artificial crosses and improvement programmes.All are destined for ornamental use.Until the 19th century,the great majority of the many ancient cultivated roses in Europe were used in perfumery and cosmetics,or had medicinal uses.Rosa damascena and Rosa centifollia are still grown and used by the French and Bulgarian perfume industries.The Asturian Massif of the Cantabrian Mountain Range provides a natural habitat for some 75%of the wild members of the genus Rosa,but until now there was no evidence that this area was home to ancient cultivated roses.A complete botanical description is here provided for a discovered ancient rose.It is also characterised according to a series of sequence tagged microsatellite sites,and its agronomic features are reported.In addition,a histological description(optical and scanning electronic microscope studies)of the petals is offered,along with an analysis of the volatile compounds present in these organs as determined by solid phase microextraction and gas chromatography-mass spectroscopy.The results reveal the uniqueness of this ancient type of rose and suggest it may be of interest to the perfume industry.

Alpine tundra species phenology is mostly driven by climate-related variables rather than by photoperiod

The study of plant phenology has frequently been used to link phenological events to various factors,such as temperature or photoperiod.In the high-alpine environment,proper timing of the phenological cycle has always been crucial to overcome harsh conditions and potential extreme events(i.e.spring frosts)but little is known about the response dynamics of the vegetation,which could shape the alpine landscape in a future of changing climate.Alpine tundra vegetation is composed by an array of species belonging to different phytosociological optima and with various survival strategies,and snowbed communities are a relevant expression of such an extreme-climate adapted flora.We set eight permanent plots with each one in a snowbed located on the Cimalegna plateau in Northwestern Italy and then we selected 10 most recurring species among our plots,all typical of the alpine tundra environment and classified in 3different pools:snowbed specialists,grassland species and rocky debris species.For 3 years we registered the phenophases of each species during the whole growing season using an adaptation of the BBCH scale.We later focused on the three most biologically relevant phenophases,i.e.,flower buds visible,full flowering,and beginning of seed dispersion.Three important season-related variables were chosen to investigate their relationship with the phenological cycle of the studied species:(i)the Day Of Year(DOY),the progressive number of days starting from the 1 st of January,used as a proxy of photoperiod,(ii)Days From Snow Melt(DFSM),selected to include the relevance of the snow dynamics,and(iii)Growing Degree Days(GDD),computed as a thermal sum.Our analysis highlighted that phenological development correlated better with DFSM and GDD than with DOY.Indeed,models showed that DOY was always a worse predictor since it failed to overcome interannual variations,while DFSM and marginally GDD were better suited to predict the phenological development of most of the species,despite differences intemperature and snowmelt date among the three years.Even if the response pattern to the three variables was mainly consistent for all the species,the timing of their phenological response was different.Indeed,species such as Salix herbacea and Ranunculus glacialis were always earlier in the achievement of the phenophases,while Agrostis rupestris and Euphrasia minima developed later and the remaining species showed an intermediate behavior.However,we did not detect significant differences among the three functional pools of species.

Modelling Manure Production in Beef Calves: Development, Evaluation, and Application of a Complete vs. Simplified Prediction Model

There has been increased interest in quantifying the manure production of livestock, primarily driven by public authorities, who aim to evaluate the environmental impact of livestock production, but also at the farm level, to manage manure storage and availability of fertilizer for crop production. Moreover, current manure production estimates from intensively reared beef calves are higher than actual production due to changes in farming systems, advances in animal genetics and feed efficiency. This study aims to redefine and update manure production estimates in intensively reared beef calves to predict manure production as a policy and planning tool, as there are no current models available. A trial was conducted to collect data on manure production during the growing-finishing period (243 d) of 54 Limousine calves (from 346.7 to 674.0 kg live weight, LW). Such data were used to develop two models to predict manure excretion: (1) a complex mechanistic model (CompM), and (2) a simplified empirical model (SimpM). Both models were evaluated against an independent dataset including a total of 4,692 animals on 31 farms and 5 breeds. Results from CompM require interpretation because the model does not output a single value but a range of manure production (minimum, medium and maximum), and would therefore be more suitable for professional use. The SimpM could be considered simple, reliable, and versatile for predicting manure excretion at farm level. SimpM could be refined and improved by including data from other studies on beef cattle with distinct characteristics and management.

Biological and molecular interplay between two viruses and powdery and downy mildews in two grapevine cultivars

Grapevine may be affected simultaneously by several pathogens whose complex interplay is largely unknown.We studied the effects of infection by two grapevine viruses on powdery mildew and downy mildew development and the molecular modifications induced in grapevines by their multiple interactions.Grapevine fanleaf virus(GFLV)and grapevine rupestris stem pitting-associated virus(GRSPaV)were transmitted by in vitro-grafting to Vitis vinifera cv Nebbiolo and Chardonnay virus-free plantlets regenerated by somatic embryogenesis.Grapevines were then artificially inoculated in the greenhouse with either Plasmopara viticola or Erysiphe necator spores.GFLV-infected plants showed a reduction in severity of the diseases caused by powdery and downy mildews in comparison to virus-free plants.GFLV induced the overexpression of stilbene synthase genes,pathogenesis-related proteins,and influenced the genes involved in carbohydrate metabolism in grapevine.These transcriptional changes suggest improved innate plant immunity,which makes the GFLV-infected grapevines less susceptible to other biotic attacks.This,however,cannot be extrapolated to GRSPaV as it was unable to promote protection against the fungal/oomycete pathogens.In these multiple interactions,the grapevine genotype seemed to have a crucial role:in‘Nebbiolo’,the virus-induced molecular changes were different from those observed in‘Chardonnay’,suggesting that different metabolic pathways may be involved in protection against fungal/oomycete pathogens.These results indicate that complex interactions do exist between grapevine and its different pathogens and represent the first study on a topic that still is largely unexplored.

Molecular memory of Flavescence dorée phytoplasma in recovering grapevines

Flavescence dorée(FD)is a destructive phytoplasma disease of European grapevines.Spontaneous and cultivardependent recovery(REC)may occur in the field in FD-infected vines starting the year following the first symptoms.However,the biological underpinnings of this process are still largely unexplored.In this study,transcriptome sequencing(RNAseq),whole-genome bisulphite sequencing(WGBS)and metabolite analysis were combined to dissect molecular and metabolic changes associated to FD and REC in leaf veins collected in the field from healthy(H),FD and REC plants of the highly susceptible Vitis vinifera‘Barbera’.Genes involved in flavonoid biosynthesis,carbohydrate metabolism and stress responses were overexpressed in FD conditions,whereas transcripts linked to hormone and stilbene metabolisms were upregulated in REC vines.Accumulation patterns of abscisic acid and stilbenoid compounds analysed in the same samples confirmed the RNAseq data.In recovery conditions,we also observed the persistence of some FD-induced expression changes concerning inhibition of photosynthetic processes and stress responses.Several differentially expressed genes tied to those pathways also underwent post-transcriptional regulation by microRNAs,as outlined by merging our transcriptomic data set with a previously conducted smallRNAseq analysis.Investigations by WGBS analysis also revealed different DNA methylation marks between REC and H leaves,occurring within the promoters of genes tied to photosynthesis and secondary metabolism.The results allowed us to advance the existence of a“molecular memory”of FDp infection,involving alterations in the DNA methylation status of REC plants potentially related to transcriptional reprogramming events,in turn triggering changes in hormonal and secondary metabolite profiles.

Quantifying the contribution of the root system of alpine vegetation in the soil aggregate stability of moraine

One fifth of the world's population is living in mountains or in their surrounding areas.This anthropogenic pressure continues to grow with the increasing number of settlements,especially in areas connected to touristic activities,such as the Italian Alps.The process of soil formation on high mountains is particularly slow and these soils are particularly vulnerable to soil degradation.In alpine regions,extreme meteorological events are increasingly frequent due to climate change,speeding up the process of soil degradation and increasing the number of severe erosion processes,shallow landslides and debris flows.Vegetation cover plays a crucial role in the stabilization of mountain soils thereby reducing the risk of natural hazards effecting downslope areas.Soil aggregate stability is one of the main soil properties that can be linked to soil loss processes.Soils developed on moraines in recently deglaciated areas typically have low levels of soil aggregation,and a limited or discontinuous vegetation cover making them more susceptible to degradation.However,soil structure can be influenced by the root system of the vegetation.Roots are actively involved in the formation of water-stable soil aggregation,increasing the stability of the soil and its nutrient content.In the present study,we aim to quantify the effect of the root system of alpine vegetation on the soil aggregate stability of the forefield of the Lys glacier,in the Aosta Valley(NW-Italy).This proglacial area provides the opportunity to study how the root system of ten pioneer alpine species from different successional stages can contribute to soil development and soil stabilization.To quantify the aggregate stability of root permeated soils,a modified wet sieving method was employed.The root length per soil volume of the different species was also determined and later correlated with the aggregate stability results.The results showed that soil aggregate stability was significantly increased by the presence of roots.The lowest soil aggregate stability was found with Epilobium fleischeri followed by Minuartia recurva and Leucanthemopsis alpina.The highest aggregate stability was found with the graminoid species.These results show a close relationship between the development of root systems of the studied species and soil aggregate stability,a factor which can be taken into consideration in order to improve the accuracy of existing susceptibility mapping for early warning and civilian protection.

Computer-assisted sperm morphometry fluorescence-based analysis has potential to determine progeny sex

This study was designed to determine the ability of computer-assisted sperm morphometry analysis （CASA-Morph） with fluorescence to discriminate between spermatozoa carrying different sex chromosomes from the nuclear morphometrics generated and different statistical procedures in the bovine species. The study was divided into two experiments. The first was to study the morphometric differences between X- and Y-chromosome-bearing spermatozoa （SX and SY, respectively）. Spermatozoa from eight bulls were processed to assess simultaneously the sex chromosome by FISH and sperm morphometry by fluorescence-based CASA-Morph. SX cells were larger than SY cells on average （P 〈 0.001） although with important differences between bulls. A simultaneous evaluation of all the measured features by discriminant analysis revealed that nuclear area and average fluorescence intensity were the variables selected by stepwise discriminant function analysis as the best discriminators between SX and SY. In the second experiment, the sperm nuclear morphometric results from CASA-Morph in nonsexed （mixed SX and SY） and sexed （SX） semen samples from four bulls were compared. FISH allowed a successful classification of spermatozoa according to their sex chromosome content. X-sexed spermatozoa displayed a larger size and fluorescence intensity than nonsexed spermatozoa （P 〈 0.05）. We conclude that the CASA-Morph fluorescence-based method has the potential to find differences between X- and Y-chromosome-bearing spermatozoa in bovine species although more studies are needed to increase the precision of sex determination by this technique.