Whole genome scanning of a Mediterranean basin hotspot collection provides new insights into olive tree biodiversity and biology.

Olive tree (Olea europaea L. subsp. europaea var. europaea) is one of the most important species of the Mediterranean region and one of the most ancient species domesticated. The availability of whole genome assemblies and annotations of olive tree cultivars and oleaster (O. europaea subsp. europaea var. sylvestris) has contributed to a better understanding of genetic and genomic differences between olive tree cultivars. However, compared to other plant species there is still a lack of genomic resources for olive tree populations that span the entire Mediterranean region. In the present study we developed the most complete genomic variation map and the most comprehensive catalog/resource of molecular variation to date for 89 olive tree genotypes originating from the entire Mediterranean basin, revealing the genetic diversity of this commercially significant crop tree and explaining the divergence/similarity among different variants. Additionally, the monumental ancient tree 'Throuba Naxos' was studied to characterize the potential origin or routes of olive tree domestication. Several candidate genes known to be associated with key agronomic traits, including olive oil quality and fruit yield, were uncovered by a selective sweep scan to be under selection pressure on all olive tree chromosomes. To further exploit the genomic and phenotypic resources obtained from the current work, genome-wide association analyses were performed for 23 morphological and two agronomic traits. Significant associations were detected for eight traits that provide valuable candidates for fruit tree breeding and for deeper understanding of olive tree biology.

Characterization of the olive endophytic community in genotypes displaying a contrasting response to Xylella fastidiosa.

BACKGROUND: Endophytes mediate the interactions between plants and other microorganisms, and the functional aspects of interactions between endophytes and their host that support plant-growth promotion and tolerance to stresses signify the ecological relevance of the endosphere microbiome. In this work, we studied the bacterial and fungal endophytic communities of olive tree (Olea europaea L.) asymptomatic or low symptomatic genotypes sampled in groves heavily compromised by Xylella fastidiosa subsp. pauca, aiming to characterize microbiota in genotypes displaying differential response to the pathogen. RESULTS: The relationships between bacterial and fungal genera were analyzed both separately and together, in order to investigate the intricate correlations between the identified Operational Taxonomic Units (OTUs). Results suggested a dominant role of the fungal endophytic community compared to the bacterial one, and highlighted specific microbial taxa only associated with asymptomatic or low symptomatic genotypes. In addition, they indicated the occurrence of well-adapted genetic resources surviving after years of pathogen pressure in association with microorganisms such as Burkholderia, Quambalaria, Phaffia and Rhodotorula. CONCLUSIONS: This is the first study to overview endophytic communities associated with several putatively resistant olive genotypes in areas under high X. fastidiosa inoculum pressure. Identifying these negatively correlated genera can offer valuable insights into the potential antagonistic microbial resources and their possible development as biocontrol agents.

Improving plant adaptation to soil antimony contamination: the synergistic contribution of arbuscular mycorrhizal fungus and olive mill waste.

BACKGROUND: This study aimed to investigate the alterations in biochemical and physiological responses of oat plants exposed to antimony (Sb) contamination in soil. Specifically, we evaluated the effectiveness of an arbuscular mycorrhizal fungus (AMF) and olive mill waste (OMW) in mitigating the effects of Sb contamination. The soil was treated with a commercial strain of AMF (Rhizophagus irregularis) and OMW (4% w/w) under two different levels of Sb (0 and 1500 mg kg-1 soil). RESULTS: The combined treatment (OMW + AMF) enhanced the photosynthetic rate (+ 40%) and chlorophyll a (+ 91%) and chlorophyll b (+ 50%) content under Sb condition, which in turn induced more biomass production (+ 67-78%) compared to the contaminated control plants. More photosynthesis in OMW + AMF-treated plants gives a route for phenylalanine amino acid synthesis (+ 69%), which is used as a precursor for the biosynthesis of secondary metabolites, including flavonoids (+ 110%), polyphenols (+ 26%), and anthocyanins (+ 63%) compared to control plants. More activation of phenylalanine ammonia-lyase (+ 38%) and chalcone synthase (+ 26%) enzymes in OMW + AMF-treated plants under Sb stress indicated the activation of phenylpropanoid pathways in antioxidant metabolites biosynthesis. There was also improved shifting of antioxidant enzyme activities in the ASC/GSH and catalytic pathways in plants in response to OMW + AMF and Sb contamination, remarkably reducing oxidative damage markers. CONCLUSIONS: While individual applications of OMW and AMF also demonstrated some degree of plant tolerance induction, the combined presence of AMF with OMW supplementation significantly enhanced plant biomass production and adaptability to oxidative stress induced by soil Sb contamination.

Disclosing the molecular basis of salinity priming in olive trees using proteogenomic model discovery.

Plant responses to salinity are becoming increasingly understood, however, salt priming mechanisms remain unclear, especially in perennial fruit trees. Herein, we showed that low-salt pre-exposure primes olive (Olea europaea) plants against high salinity stress. We then performed a proteogenomic study to characterize priming responses in olive roots and leaves. Integration of transcriptomic and proteomic data along with metabolic data revealed robust salinity changes that exhibit distinct or overlapping patterns in olive tissues, among which we focused on sugar regulation. Using the multi-crossed -omics data set, we showed that major differences between primed and nonprimed tissues are mainly associated with hormone signaling and defense-related interactions. We identified multiple genes and proteins, including known and putative regulators, that reported significant proteomic and transcriptomic changes between primed and nonprimed plants. Evidence also supported the notion that protein post-translational modifications, notably phosphorylations, carbonylations and S-nitrosylations, promote salt priming. The proteome and transcriptome abundance atlas uncovered alterations between mRNA and protein quantities within tissues and salinity conditions. Proteogenomic-driven causal model discovery also unveiled key interaction networks involved in salt priming. Data generated in this study are important resources for understanding salt priming in olive tree and facilitating proteogenomic research in plant physiology.

Physiological and biochemical study of the drought tolerance of 14 main olive cultivars in the Mediterranean basin.

A complete study of 14 olive cultivars of great economic importance was carried out. These cultivars are Arbequina, Arbosana, Chemlali, Cornicabra, Cornezuelo de Jaén, Empeltre, Frantoio, Hojiblanca, Koroneiki, Manzanilla de Sevilla, Martina, Picual, Sikitita1 and Sikitita 2. All of them are certified by the World Olive Germplasm Bank of Córdoba (Spain). They are predominant cultivars in the olive groves of different locations throughout the Mediterranean basin, and they were subjected to total water deficit for a minimum of 14 days and a maximum of 42 days in the present study. Data such as chlorophyll content, soil moisture and specific leaf area were gathered. Photosynthetic parameters measured at the respective saturation irradiance of each cultivar were also analysed: assimilation rate, transpiration, stomatal conductance, photosynthetic efficiency, photochemical and non-photochemical quenching, photonic flux density, electron transference ratio, efficient use of water and amount of proline and malondialdehyde as indicators of oxidative stress. In addition to the control, two different experimental conditions were analysed: moderate drought, after 14 days of lack of irrigation, and severe drought, after 28-42 days of total absence of irrigation, depending on the tolerance of each cultivar. Based on the results, the cultivars were characterised and divided into four groups according to their drought tolerance: tolerant, moderately tolerant, moderately sensitive and sensitive to drought. This work represents the first contribution of drought tolerance of a considerable number of olive cultivars, with all of them being subjected to the same criteria and experimental conditions for their classification.

Olive reproductive biology: implications for yield, compatibility conundrum, and environmental constraints.

Olive (Olea europaea L.) is an important Mediterranean tree species with a longstanding history of cultivation, boasting a diverse array of local cultivars. While traditional olive orchards are valued for their cultural and aesthetic significance, they often face economic sustainability challenges in the modern context. The success of both traditional and newly introduced cultivars (e.g. those obtained by cross-breeding) is hindered by self-incompatibility, a prevalent issue for this species that results in low fruit set when limited genetic diversity is present. Further, biological, environmental, and agronomic factors have been shown to interlink in shaping fertilization patterns, hence impacting on the final yield. Climatic conditions during pollination, such as excessive rainfall or high temperatures, can further exacerbate the problem. In this work, we provide an overview of the various factors that trigger the phenomenon of suboptimal fruit set in olive trees. This work provides a comprehensive understanding of the interplay among these factors, shedding light on potential mechanisms and pathways that contribute to the observed outcomes in the context of self-incompatibility in olive.

Investigating the in vivo biocontrol and growth-promoting efficacy of Bacillus sp. and Pseudomonas fluorescens against olive knot disease.

Olive knot disease, caused by Pseudomonas savastanoi, poses a significant threat to olive cultivation, necessitating sustainable alternatives to conventional chemical control. This study investigates the biocontrol effectiveness of Bacillus sp. (Og2) and Pseudomonas fluorescens (Oq5), alone and combined, against olive knot disease. Olive plants were sprayed with 5 ml of the bacteria until uniformly wet, with additional application to the soil surface. Pathogen injection occurred 24 h later. The results revealed that treating plants with a combination of both bacteria provided the highest reduction in disease severity (89.58 %), followed by P. fluorescens alone (69.38 %). Significant improvements were observed in shoot height, particularly with the combination of Bacillus sp. and P. fluorescens. The root length of olive seedlings treated with P. fluorescens and Bacillus sp., either alone or in combination, was significantly longer compared to the control and pathogen-treated seedlings. In terms of root dry weight, the most effective treatments were treated with P. fluorescens was the highest (82.94 g) among all treatments followed by the combination of both isolates with seedlings inoculated with P. savastanoi. These findings underscore the potential of Bacillus sp. and Pseudomonas fluorescens as effective biocontrol agents against olive knot disease and promoting olive seedlings growth, providing a sustainable and environmentally friendly approach to disease management.

The novel bacteriocin BacYB1 produced by Leuconostoc mesenteroides YB1: From recent analytical characterization to biocontrol Verticillium dahliae and Agrobacterium tumefaciens.

Biocontrol of phytopathogens involving the use of bioactive compounds produced by lactic acid bacteria (LAB), is a promising approach to manage many diseases in agriculture. In this study, a lactic acid bacterium designated YB1 was isolated from fermented olives and selected for its antagonistic activity against Verticillium dahliae (V. dahliae) and Agrobacterium tumefaciens (A. tumefaciens). Based on the 16S rRNA gene nucleotide sequence analysis (1565 pb, accession number: OR714267), the new isolate YB1 bacterium was assigned as Leuconostoc mesenteroides YB1 (OR714267) strain. This bacterium produces an active peptide "bacteriocin" called BacYB1, which was purified in four steps. Matrix-assisted lasers desorption/ionization (MALDI) time-of-flight (TOF) mass spectrometry (MS) based approach was performed to identify and characterize BacYB1. The exact mass was 5470.75 Da, and the analysis of the N-terminal sequence (VTRASGASTPPGTASPFKTL) of BacYB1 revealed no significant similarity to currently available antimicrobial peptides. The BacYB1 displayed a bactericidal mode of action against A. tumefaciens. The potentiel role of BacYB1 to supress the growth of A. tumefaciens was confirmed by live-dead cells viability assay. In pot experiments, the biocontrol efficacy of BacYB1 against V. dahliae wilt on young olive trees was studied. The percentage of dead plants (PDP) and the final mean symptomes severity (FMS) of plants articifialy infected by V. dahliae and treated with the pre-purified peptide BacYB1 (preventive and curative treatments) were significantly inferior to untreated plants. Biochemical analysis of leaves of the plants has shown that polyophenols contents were highly detected in plants infected by V. dahliae and the highest contents of chlorophyl a, b and total chlorophyll were recorded in plants treated with the combination of BacYB1 with the biofertilisant Humivital. BacYB1 presents a promising alternative for the control of Verticillium wilt and crown gall diseases.

Investigation on screening, identification, and fermentation characteristics of Yunnan olive in the fermented liquid utilizing five strains of Saccharomyces cerevisiae.

Refined indigenous Saccharomyces cerevisiae can enhance refinement, sophistication, and subtlety of fruit wines by showcasing exceptional regional characteristics. In order to identify exceptional indigenous S. cerevisiae strains from Yunnan olive, this study isolated 60 yeast strains from wild Yunnan olive fermentation mash. The five S. cerevisiae strains were subjected to morphological and molecular biological identification, followed by evaluation of their fermentation performance, ethanol production capacity, ester production capacity, H2S production capacity, killing capacity, and tolerance. Strains LJM-4, LJM-10, and LJM-26 exhibited robust tolerance to 6% ethanol volume fraction, pH 2.8, sucrose concentration of 400 g/L, SO2 concentration of 0.3 g/L, glucose concentration of 400 g/L at both 40 °C and 15 °C. Additionally, strain LJM-10 demonstrated a faster fermentation rate compared to the other strains. Among the tested S. cerevisiae strains evaluated in this study for olive wine fermentation process in Yunnan region; strain LJM-10 displayed superior abilities in terms of ester and ethanol production while exhibiting the lowest H2S production levels. These findings suggest that strain LJM-10 holds great potential as an excellent candidate for optimizing fruit wine S. cerevisiae fermentation processes in Yunnan olive fruit wine.

A mathematical model of biofilm growth and spread within plant xylem: Case study of Xylella fastidiosa in olive trees.

Xylem-limited bacterial pathogens cause some of the most destructive plant diseases. Though imposed measures to control these pathogens are generally ineffective, even among susceptible taxa, some hosts can limit bacterial loads and symptom expression. Mechanisms by which this resistance is achieved are poorly understood. In particular, it is still unknown how differences in vascular structure may influence biofilm growth and spread within a host. To address this, we developed a novel theoretical framework to describe biofilm behaviour within xylem vessels, adopting a polymer-based modelling approach. We then parameterised the model to investigate the relevance of xylem vessel diameters on Xylella fastidiosa resistance among olive cultivars. The functionality of all vessels was severely reduced under infection, with hydraulic flow reductions of 2-3 orders of magnitude. However, results suggest wider vessels act as biofilm incubators; allowing biofilms to develop over a long time while still transporting them through the vasculature. By contrast, thinner vessels become blocked much earlier, limiting biofilm spread. Using experimental data on vessel diameter distributions, we were able to determine that a mechanism of resistance in the olive cultivar Leccino is a relatively low abundance of the widest vessels, limiting X. fastidiosa spread.

Using 'sentinel' plants to improve early detection of invasive plant pathogens.

Infectious diseases of plants present an ongoing and increasing threat to international biosecurity, with wide-ranging implications. An important challenge in plant disease management is achieving early detection of invading pathogens, which requires effective surveillance through the implementation of appropriate monitoring programmes. However, when monitoring relies on visual inspection as a means of detection, surveillance is often hindered by a long incubation period (delay from infection to symptom onset) during which plants may be infectious but not displaying visible symptoms. 'Sentinel' plants-alternative susceptible host species that display visible symptoms of infection more rapidly-could be introduced to at-risk populations and included in monitoring programmes to act as early warning beacons for infection. However, while sentinel hosts exhibit faster disease progression and so allow pathogens to be detected earlier, this often comes at a cost: faster disease progression typically promotes earlier onward transmission. Here, we construct a computational model of pathogen transmission to explore this trade-off and investigate how including sentinel plants in monitoring programmes could facilitate earlier detection of invasive plant pathogens. Using Xylella fastidiosa infection in Olea europaea (European olive) as a current high profile case study, for which Catharanthus roseus (Madagascan periwinkle) is a candidate sentinel host, we apply a Bayesian optimisation algorithm to determine the optimal number of sentinel hosts to introduce for a given sampling effort, as well as the optimal division of limited surveillance resources between crop and sentinel plants. Our results demonstrate that including sentinel plants in monitoring programmes can reduce the expected prevalence of infection upon outbreak detection substantially, increasing the feasibility of local outbreak containment.

Unveiling the silver lining: examining the effects of biogenic silver nanoparticles on the growth dynamics of in vitro olive shoots.

The current study aimed to evaluate the effects of biogenic silver nanoparticles (AgNPs) on growth behavior and leaf anatomy of in vitro growing shoots of 'Picual' and 'Dolce' olive cultivars. Biosynthesis of AgNPs was carried out using the cell-free filtrate of Fusarium oxysporum. The dimension and shape of the synthesized AgNPs have been analyzed using spectroscopy and topography analysis tools, confirming that the biosynthesis of AgNPs is a crystalline nanostructure with an average particle size of 37 nm. The shoots of the selected olive cultivars were cultured on Rugini olive medium-supplemented AgNPs at 0, 10, 20, and 30mg L- 1. The effect of genotypes on shoot multiplication was significant, 'Picual' recorded higher values of shoot growth parameters compared with 'Dolce' cultivar. Adding AgNPs to the culture medium significantly affected the growth of in vitro olive shoots. AgNPs at 20 and 30mg L- 1 produced higher values of the number of shoots, shoot length, and leaf number of Picual cv. compared with the control treatments, but the higher AgNPs concentration harmed the growth parameters of Dolce cv. and recorded lower growth values compared with the lower concentration (10mg L- 1). AgNPs had a significant effect on leaf morphology and their anatomical structure. The current results showed that the stimulatory effect of AgNPs on shoot growth of in vitro olive shoots is highly dependent on plant genotype and nanoparticle concentration.

Integrated Transcriptome and Metabolome Analysis Revealed the Key Role of the Flavonoid Biosynthesis in Olive Defense Against Alternaria alternata.

The olive tree is an important oil woody plant with high economic value, yet it is vulnerable to the attack of numerous fungi. The successful control of olive fungal diseases requires a comprehensive understanding of the disease resistance mechanisms in plants. Here, we isolated Alternaria alternata from the diseased leaves of olive plants, and screened a resistant ("Leccino") and susceptible ("Manzanilla de Sevilla") cultivar from eight olive cultivars to explore their resistance mechanisms. Transcriptomic and metabolomic analyses identified the flavonoid biosynthesis as a key defense pathway against A. alternata. Five important transcription factors associated with flavonoid biosynthesis were also determined. The overexpression of OeWRKY40 significantly enhanced the disease resistance of the susceptible cultivar and upregulated the expression of genes involved in flavonoid biosynthesis and the accumulation of related metabolites. LUC assays further proved that OeWRKY40 can activate the expression of OeC4H. These results help to better clarify the molecular mechanisms of flavonoid biosynthesis against A. alternata. Our study provides key information for further exploration of the molecular pathways of olive plants and their resistance to fungi, an important factor for molecular breeding and utilization of resistant cultivars.

Anti-inflammatory effects of Olive (olea europaea L.) fruit extract in LPS-stimulated RAW264.7 cells via MAPK and NF-κB signal pathways.

BACKGROUND: Olive is an evergreen tree of Oleaceae Olea with numerous bioactive components. While the anti-inflammatory properties of olive oil and the derivatives are well-documented, there remains a dearth of in-depth researches on the immunosuppressive effects of olive fruit water extract. This study aimed to elucidate the dose-effect relationship and underlying molecular mechanisms of olive fruit extract in mediating anti-inflammatory responses. METHODS AND RESULTS: The impacts of olive fruit extract on the release of nitric oxide (NO), tumor necrosis factor (TNF-α), interleukins-6 (IL-6) and reactive oxygen species (ROS) were assessed in RAW264.7 cells induced by lipopolysaccharide (LPS). For deeper understanding, the expression of genes encoding inducible nitric oxide synthase (iNOS), cyclooxygenase-2 (COX-2), TNF-α and IL-6 was quantitatively tested. Additionally, the expression patterns of MAPK and NF-κB pathways were further observed to analyze the action mechanisms. Results suggested that olive fruit extract (200, 500, 1000 µg/mL) markedly exhibited a dose-dependent reduction in the generation of NO, TNF-α, IL-6 and ROS, as well as the expression of correlative genes studied. The activation of ERK, JNK, p38, IκB-α and p65 were all suppressed when p65 nuclear translocation was further restricted by olive fruit extract in NF-κB and MAPK signal pathways. CONCLUSIONS: Olive fruit extract targeted imposing restrictions on the signal transduction of key proteins in NF-κB and MAPK pathways, and thereby lowered the level of inflammatory mediators, which put an enormous hindrance to inflammatory development. Accordingly, it is reasonable to consider olive fruit as a potent ingredient in immunomodulatory products.

Rhizosphere-xylem sap connections in the olive tree microbiome: implications for biostimulation approaches.

AIMS: Climate change is endangering olive groves. Farmers are adapting by exploring new varieties of olive trees and examining the role of microbiomes in plant health.The main objectives of this work were to determine the primary factors that influence the microbiome of olive trees and to analyze the connection between the rhizosphere and endosphere compartments. METHODS AND RESULTS: The rhizosphere and xylem sap microbiomes of two olive tree varieties were characterized by next-generation 16S rRNA amplicon sequencing, and soil descriptors were analyzed. Bacterial communities in the rhizosphere of olive trees were more diverse than those found in the xylem sap. Pseudomonadota, Actinobacteriota, Acidobacteriota, and Bacillota were the dominant phyla in both compartments. At the genus level, only very few taxa were shared between soil and sap bacterial communities. CONCLUSIONS: The composition of the bacteriome was more affected by the plant compartment than by the olive cultivar or soil properties, and a direct route from the rhizosphere to the endosphere could not be confirmed. The large number of plant growth-promoting bacteria found in both compartments provides promising prospects for improving agricultural outcomes through microbiome engineering.

Rhizobacterial diversity of Portuguese olive cultivars in the Douro valley and their potential as plant growth promoters.

AIMS: This study investigated the bacterial communities in the rhizosphere of two traditional Portuguese olive cultivars, Cobrançosa and Negrinha de Freixo, in relation to soil properties. Additionally, we aimed to isolate and identify bacteria with potential for biocontrol and other plant growth-promoting traits from these rhizosphere communities. METHODS AND RESULTS: Bacterial communities in the olive rhizosphere were investigated using a metabarcoding approach and the soil physicochemical properties of the olive groves were also analyzed. Higher bacterial richness was associated with Negrinha de Freixo growing in soil with high organic matter content and water-holding capacity. In contrast, the soils of the Cobrançosa grove presented higher pH and electric conductivity. Negrinha de Freixo rhizosphere was enriched with ASVs (Amplicon Sequence Variants) belonging to Bacillus, Gaiella, Acidothermus, Bradyrhizobium, and uncultured Xanthobacteraceae. On the other hand, the Cobrançosa rhizosphere was characterized by higher relative abundance of Streptomyces and Sphingomonas. Bacterial isolation from the rhizosphere and screening for plant growth-promoting activities were also performed. Six bacteria strains, predominantly Bacillus isolated from Negrinha de Freixo, demonstrated antagonistic activities against the olive fungal pathogen Colletotrichum gloeosporoides and other plant growth promotion (PGP) traits. CONCLUSIONS: Our findings demonstrate that the structure of rhizosphere bacterial communities associated with olive trees is shaped by both plant cultivar and soil-related factors. The higher number of bacterial species in the rhizosphere of Negrinha de Freixo was related to a higher organic matter content and a greater abundance of isolates with plant growth promotion traits, particularly Bacillus strains.

Investigating the characteristics of biomass wastes via particle feeder in downdraft gasifier.

Particle feeding plays a crucial role in the gasifier due to its effects on the efficiency and performance metrics of the thermochemical process. Investigating particle size distribution's impact on downdraft gasification reactor performance, this study delves into the significance of feedstock characteristics (moisture, volatile matter, fixed carbon, and ash contents) during the particle feeding stage. Various biomass wastes (date palm waste, olive pomace and sewage sludge) at diverse compositions and sizes are subjected to empirical determination of mass flow rates (MFR), power ratings, and storage times for each feedstock. The preheating process in the gasifier is considered, employing both an approximation and analytical solution. In addition, the influence of the equivalence ratio (ER) on the syngas yield is analyzed. The collected data reveals that for average particle size of 200 µm, the highest MFR (in g/min) are 0.518 ± 0.033, 7.691 ± 0.415, and 16.111 ± 1.050, for palm wood biomass, olive pomace and sewage sludge, respectively. Smaller particles (80 µm) led to extended storage times. Moreover, the lumped capacitance approximation method consistently underestimates preheating time, with a percentage error of 6.26%-17.08%. Response surface methodology (RSM) optimization analysis provides optimal gasification conditions for palm wood biomass, olive pomace, and sewage sludge with maximum cold gas efficiencies (CGEs) of 58.01%, 63.29%, and 52.27%. The peak conversion was attained at gasification temperatures of 1089.83 °C, 1151.93 °C, and 1102.91 °C for palm wood biomass, olive pomace, and sewage sludge, respectively. In addition, gasification equilibrium model determined optimal gasification temperatures as 1150 °C for palm biomass, 1200 °C for olive pomace, and 1150 °C for sewage sludge with respective syngas efficiencies of 59.62%, 64.13%, and 53.66%. Consequently, the examination of the dosing procedure, preheating dynamics, particle dimensions, ER, storage time, and their combined impacts offer practical insights to effectively control downdraft gasifiers in handling a variety of feedstocks.

Acid insoluble lignin material production by chemical activation of olive endocarps for an efficient furfural adsorption-removal from aqueous solutions.

The present work describes a protocol of chemical activation, with acid catalyst, of olive endocarps to obtain acid insoluble lignin-rich materials with high capacities for the adsorption of furfural present in aqueous media. During biomass activation, factors such as acid concentration, reaction time and temperature, solid/liquid ratio and the presence of water extractives strongly affected both the surface characteristics of the treated bioadsorbents and their capacities for furfural retention (percentage increase close to 600% with respect to the crude biomass). Once a treated solid with good adsorbent properties was obtained, the optimal conditions for adsorption were found: stirring speed 80 rpm, temperature 303 K and adsorbent load 7.5 g solid/50 cm3. Kinetic study indicated the pseudo-second order model provided the best fit of the experimental data. At 303 K, the equilibrium adsorption capacities values ranged from 2.27 mg g-1 to 29.29 mg g-1, for initial furfural concentrations between 0.49 g dm-3 and 12.88 g dm-3. Freundlich model presented the best isotherm (R2 = 0.996 and SE = 4.7%) providing KF and n values of 0.115 (mg g-1) (mg dm-3)-n and 0.610, respectively. Since physical interactions predominate in the adsorption of furfural on chemically activated olive endocarps, the furfural removal process could have occurred reversibly on the heterogeneous surface of the bioadsorbents.

2G-lactic acid from olive oil supply chain waste: olive leaves upcycling via Lactobacillus casei fermentation.

The transition towards a sustainable model, particularly the circular economy, emphasizes the importance of redefining waste as a valuable resource, paving the way for innovative upcycling strategies. The olive oil industry, with its significant output of agricultural waste, offers a promising avenue for high-value biomass conversion into useful products through microbial processes. This study focuses on exploring new, high-value applications for olive leaves waste, utilizing a biotechnological approach with Lactobacillus casei for the production of second-generation lactic acid. Contrary to initial expectations, the inherent high polyphenol content and low fermentable glucose levels in olive leaves posed challenges for fermentation. Addressing this, an enzymatic hydrolysis step, following a preliminary extraction process, was implemented to increase glucose availability. Subsequent small-scale fermentation tests were conducted with and without nutrient supplements, identifying the medium that yielded the highest lactic acid production for scale-up. The scaled-up batch fermentation process achieved an enhanced conversion rate (83.58%) and specific productivity (0.26 g/L·h). This research confirms the feasibility of repurposing olive waste leaves for the production of lactic acid, contributing to the advancement of a greener economy through the valorization of agricultural waste. KEY POINTS: • Olive leaves slurry as it did not allow L. casei to ferment. • High concentrations of polyphenols inhibit fermentation of L. casei. • Enzymatic hydrolysis combined to organosolv extraction is the best pretreatment for lactic acid production starting from leaves and olive pruning waste.

Efficacy of supplementing Aspergillus awamori in enhancing growth performance, gut microbiota, digestibility, immunity, and antioxidant activity of heat-stressed broiler chickens fed diets containing olive pulp.

BACKGROUND: Gut microbes play a significant role in digestion, developing immunity, and intestinal health. Therefore, direct-fed microbials are used to modify gut microbiota, maintain a healthy digestive system, enhance immunity, and promote the broilers' performance. In addition, it has a role in improving the utilization of unconventional feed ingredients (olive pulp, OP). This study provides the potential role of Aspergillus awamori in enhancing gut microbial content, nutrient utilization, growth performance, and antioxidative status in heat-stressed broiler chickens fed diets containing olive pulp. METHODS: Three hundred chicks (Ross 308; one day old) were divided into four treatment groups (75 chick/ group) randomly, as follows; CON: chicks fed a basal diet based on corn and soybean meal, OP10: chicks fed a diet containing 10% OP, OA1: chicks fed a diet containing OP with A. awamori at 100 mg per kg, OA2: chicks fed a diet containing OP with A. awamori at 200 mg per kg. RESULTS: Adding A. awamori to the broiler diet that contains OP had a positive effect on productive performance via enhancing nutrition digestibility, body weight gain, feed conversion ratio, and carcass characteristics. A. awamori supplementation had a positive impact on immune responses by increasing serum immunoglobulin G and the relative weight of bursa of Fabricius (P < 0.05) compared to the other groups. Chickens fed A. awamori showed a noticeable improvement in the oxidative status through the increase in the level of serum superoxide dismutase, and glutathione peroxidase, and the decrease in the level of malondialdehyde. Feeding A. awamori also modified the intestinal microbial content by increasing the population of Lactobacillus (P < 0.05). CONCLUSIONS: Our study indicated that adding 200 mg A. awamori reduced the negative effect of heat stress by modifying the microbial content of the intestine, immune response, and enhancing feed utilization, thus improving broiler performance, as well as, improving the nutritional value of the olive pulp. Therefore, adding A. awamori to the OP diet can be effectively used in heat-stressed broiler diets.