Efficacy of cognitive stimulation therapy virtual program for older adults with dementia in COVID-19 isolation.

BACKGROUND: Cognitive stimulation virtual therapy (CSVT) is an evidence-based psychosocial intervention for people with mild-to-moderate dementia due to various etiological factors. OBJECTIVE: The aim of the present study was to assess the efficacy of a cognitive stimulation virtual therapy CSVT program, in individuals who have vascular or Alzheimer dementia in COVID-19 isolation. METHODS: Older adults with mild vascular or Alzheimer dementia (N = 20) were assigned to one of two programs: one group (N = 10) attended during six months, two sessions per week program of the cognitive stimulation virtual therapy CSVT program, while the other, active control group (N = 10) took part in alternative activities. The following tests were applied to their primary caregivers. A short form of the Informant Questionnaire on Cognitive Decline in the Elderly (IQCODE) and quality of life (Quality of Life (QoL) RESULTS: Compared with the active controls, the cognitive stimulation virtual therapy CSVT program showed a greater improvement in general cognitive functioning after the intervention (i.e. score increase on the IQCODE test). A trend towards improvement was also identified in short term/working memory and perceived quality of life (Quality of life (QoL) of elderly with dementia). The primary caregivers also perceived an improvement in mood, stress, anxiety and quality of sleep after the start of the virtual therapy during COVID-19 isolation. CONCLUSION: The present results support the efficacy of cognitive stimulation virtual therapy CSVT program in people with dementia during COVID-19 isolation.

Anthropogenic particles determination in northern krill (Meganyctiphanes norvegica) from the Azores region: A keystone species for marine food webs.

Plastic pollution poses a significant threat to marine ecosystems. Microfibres from fabrics have become the most prevalent shape of microplastic found in the marine environment. The northern krill (Meganyctiphanes norvegica) is the most abundant euphausiid species in the northern hemisphere, playing a crucial role in various pelagic ecosystems. Anthropogenic microparticles in northern krill was assessed for the first time in samples collected in the Azores on two occasions - April 2019 (n = 480) and April 2023 (n = 480). Analysis of all individuals revealed 533 anthropogenic particles, with an average abundance of 0.56 ± 0.14 items per individual and, no significant differences between years. Microfibres were the most common shape (94.8 %), with the remaining items being fragments (5.2 %), and blue and black were the predominant colours. MicroFourier transform infrared spectroscopy analysis (µFTIR) of 22.1 % of the total number of particles, showed that they were mainly cellulosic (65.3 %) - either natural or semisynthetic - followed by polyester (7.6 %). Our finding of microplastics in the northern krill raises important questions due to its crucial role in marine food webs. The intake of anthropogenic particles, particularly those that are 100 % synthetic, suggests that the northern krill may act as a transfer vector of these pollutants to higher trophic levels.

Beneath the water column: Uncovering microplastic pollution in the sublittoral coastal sediments of the Canary Islands, Spain.

Marine ecosystems pollution by microplastics (MPs) is a global problem of special concern. The present study examines the prevalence and distribution of MPs and cellulosic particles in sublittoral coastal sediments of the Canary Islands archipelago (Spain). At twenty-six different locations alongside seven islands, three samples were taken parallel to the shoreline between 1 and 10 m depth (n = 78). Sediment samples were primarily digested with a H2O2 solution followed by four flotations in a saturated NaCl solution. The mean concentration obtained was 3.9 ± 1.6 items/g of dry weight. A similar distribution pattern was observed across all islands concerning particles morphology, color, size and composition: mainly colorless/translucent and blue fibers (60.0%). Additionally, fragments were also found, and to a much lesser extent microbeads, films and tangled messes. MicroFourier Transform Infrared spectroscopy analysis of 12.5% of the fibers, showed that they were mainly cellulosic (54.5%) -either natural or semisynthetic- followed by polyester (22.7%) and acrylic (4.5%). The potential correlation between particle distribution in nearshore sediments and wave intensity was also explored. This work provides the first comprehensive report on the current MPs content of the seabed of the region.

Anthropogenic debris pollution in yellow-legged gull (Larus michahellis atlantis) nests in biosphere reserves of the Canary Islands.

Anthropogenic debris, particularly plastic pollution, has emerged as a significant environmental threat to biodiversity. Given that seabirds interact with artificial debris through ingestion, entanglement, and nest incorporation, it is particularly important to quantify the quantity, origins, and chemical composition of these debris items. In this work, it was evaluated for the first time the occurrence of anthropogenic debris in nests of yellow-legged gull (Larus michahellis atlantis) in biosphere reserves of the Canary Islands (Spain). A total of 48 abandoned nests were collected from five remote and hardly accessible sampling areas, revealing that 81.3 % contained anthropogenic waste, with plastic accounting for 34.7 % of the debris, followed by metal (33.6 %) and paper (19.6 %). On average, 32.8 ± 40.9 items were found per nest. Regarding the origin, food packagings (47.8 %), personal hygiene products (21.7 %), and textiles (15.8 %) were identified as the predominant sources. Furthermore, the polymer composition of the plastics was characterised by means of Fourier-transform infrared spectroscopy analysis, being polyester the most abundant (38.2 %), followed by polyethylene (25.6 %) and rayon (10.3 %). The incorporation of anthropogenic debris into nest construction may result from outdoor human activities carried out far from nesting areas.

Nanoplastics in the soil environment: Analytical methods, occurrence, fate and ecological implications.

Soils play a very important role in ecosystems sustainability, either natural or agricultural ones, serving as an essential support for living organisms of different kinds. However, in the current context of extremely high plastic pollution, soils are highly threatened. Plastics can change the chemical and physical properties of the soils and may also affect the biota. Of particular importance is the fact that plastics can be fragmented into microplastics and, to a final extent into nanoplastics. Due to their extremely low size and high surface area, nanoplastics may even have a higher impact in soil ecosystems. Their transport through the edaphic environment is regulated by the physicochemical properties of the soil and plastic particles themselves, anthropic activities and biota interactions. Their degradation in soils is associated with a series of mechanical, photo-, thermo-, and bio-mediated transformations eventually conducive to their mineralisation. Their tiny size is precisely the main setback when it comes to sampling soils and subsequent processes for their identification and quantification, albeit pyrolysis coupled with gas chromatography-mass spectrometry and other spectroscopic techniques have proven to be useful for their analysis. Another issue as a consequence of their minuscule size lies in their uptake by plants roots and their ingestion by soil dwelling fauna, producing morphological deformations, damage to organs and physiological malfunctions, as well as the risks associated to their entrance in the food chain, although current conclusions are not always consistent and show the same pattern of effects. Thus, given the omnipresence and seriousness of the plastic menace, this review article pretends to provide a general overview of the most recent data available regarding nanoplastics determination, occurrence, fate and effects in soils, with special emphasis on their ecological implications.

Microplastics in snow of a high mountain national park: El Teide, Tenerife (Canary Islands, Spain).

Human activities have introduced high amounts of microplastics (MPs) into the atmosphere that can be transported long distances and be later deposited in terrestrial and aquatic ecosystems with precipitation (rain or snow). In this work, it has been assessed the presence of MPs in the snow of El Teide National Park (Tenerife, Canary Islands, Spain, 2150-3200 m above sea level) after two storm episodes (January-February 2021). The data set (63 samples) was divided into three groups: i) samples from "accessible areas" (after the first storm episode and in places with a strong previous/recent anthropogenic activity); ii) "pristine areas" (after the second storm episode, in places with no previous anthropogenic activity), and iii) "climbing areas" (after the second storm episode, in places with a soft recent anthropogenic activity). Similar pattern profiles were observed among sampling sites in terms of morphology, colour and size (predominance of blue and black microfibers of 250-750 µm length), as well as in composition (predominance of cellulosic -either natural or semisynthetic-, with a 62.7 %, polyester, 20.9 %, and acrylic, 6.3 %, microfibers); however, significant differences in MPs concentrations were found between samples collected in pristine areas (average concentration of 51 ± 72 items/L) and those obtained in places with a previous anthropogenic activity (average concentration of 167 ± 104 and 188 ± 164 items/L in "accessible areas" and "climbing areas", respectively). This study shows, for the first time, the presence of MPs in snow samples from a high altitude protected area on an insular territory and suggests that the sources of these contaminants could be atmospheric transport and local human outdoor activities.

Bacterial Colonization of Microplastics at the Beaches of an Oceanic Island, Tenerife, Canary Islands.

(1) Isolated systems, such as oceanic islands, are increasingly experiencing important problems related to microplastic debris on their beaches. The formation of microbial biofilm on the surface of microplastics present in marine environments provides potential facilities for microorganisms to survive under the biofilm. Moreover, microplastics act as a vehicle for the dispersion of pathogenic organisms, constituting a new route of exposure for humans. (2) In this study, the microbial content (FIO and Vibrio spp. and Staphylococcus aureus) of microplastics (fragments and pellets) collected from seven beaches of the oceanic island of Tenerife, in the Canary Islands (Spain), was determined. (3) Results showed that Escherichia coli was present in 57.1% of the fragments and 28.5% of the pellets studied. In the case of intestinal Enterococci, 85.7% of the fragments and 57.1% of the pellets tested positive for this parameter. Finally, 100% of the fragments and 42.8% of the pellets analyzed from the different beaches contained Vibrio spp. (4) This study shows that microplastics act as reservoirs of microorganisms that can increase the presence of bacteria indicating faecal and pathogenic contamination in bathing areas.

Three-dimensional evaluation of beaches of oceanic islands as reservoirs of plastic particles in the open ocean.

The quantification of plastic debris on beaches has been extensively used as an indicator of plastic pollution in the marine environment. However, most efforts have focused on surface layers, with few investigations looking deeper into the substrate, thus underestimating total standing stocks. Such information is crucial to improve our understanding of where plastic accumulates in the oceans. In this study, we investigated the three-dimensional distribution of plastic (>1 mm) in three sandy beaches located in oceanic islands of the North Atlantic (Azores and the Canary Islands) that are known to accumulate significant quantities of small plastic debris at the surface layer. On each beach, we collected a total of 16 sediment cores down to 1 m depth, from the high tide line up to the backshore following a stratified random sampling design spread across four different levels across the beach. Samples were taken every 10 cm down to 1 m into the sand. Our results revealed the presence of plastic items in the deepest layers with subsurface layers accounting for 84 % of the total plastic abundance and with a similar pattern in terms of size, shape, colour and composition. Furthermore, we found increasing plastic concentrations towards the upper levels of the beach, indicating longer term accumulation in the backshore. Collectively, this study suggests that the plastic items reaching sandy beaches of the Macaronesia are being incorporated into its deepest layers, acting as reservoirs of plastic in the open ocean.

Fluoride Levels in Supply Water from the Canary Islands Region.

The Canary Islands, located in the Atlantic Ocean, are an archipelago of volcanic origin which, for decades, has been affected by natural fluoride contamination in the water supply of some of its islands, mainly the island of Tenerife. In addition, recent volcanic eruptions in the archipelago and the increased demand for water supply have led to an increase in the fluoride content in other areas which, historically, were not affected. Fluoride content was determined in 274 water supply samples from the most populated islands of the Canary Islands (Tenerife and Gran Canaria) collected during the months of June 2021 to May 2022. The samples were analysed by fluoride ion selective potentiometry. The highest concentrations in Tenerife were found in the municipalities of Sauzal (7.00 mg/L) and Tegueste (5.39 mg/L), both water samples are over the parametric value of 1.5 mg/L set in the supply water legislation. In the Gran Canaria Island, the highest fluoride levels were found in Valsequillo and Mogán with 1.44 mg/L in both locations, but under the parametric fluoride value abovementioned. Consumption of just 1 L of water per day in the El Sauzal area would result in a contribution rate of 77% for adults and children over 15 years of age (Upper Level value of 7 mg/day) and 108% for children 9-14 years of age (UL value of 5 mg/day). The contribution rates increase considerably, reaching or exceeding 100% of the reference value (UL) with increasing consumption of 1 to 2 L of water per day. Therefore, it is considered that there is a health risk of overexposure to fluoride on the island of Tenerife. In the case of the island of Gran Canaria, it has been shown that even the consumption of 2 litres of water per day does not confer contribution rates that pose a health risk.

A nationwide monitoring of atmospheric microplastic deposition.

Plastic production continues to increase every year, yet it is widely acknowledged that a significant portion of this material ends up in ecosystems as microplastics (MPs). Among all the environmental compartments affected by MPs, the atmosphere remains the least well-known. Here, we conducted a one-year simultaneous monitoring of atmospheric MPs deposition in ten urban areas, each with different population sizes, economic activities, and climates. The objective was to assess the role of the atmosphere in the fate of MPs by conducting a nationwide quantification of atmospheric MP deposition. To achieve this, we deployed collectors in ten different urban areas across continental Spain and the Canary Islands. We implemented a systematic sampling methodology with rigorous quality control/quality assurance, along with particle-oriented identification and quantification of anthropogenic particle deposition, which included MPs and industrially processed natural fibres. Among the sampled MPs, polyester fibres were the most abundant, followed by acrylic polymers, polypropylene, and alkyd resins. Their equivalent sizes ranged from 22 µm to 398 µm, with a median value of 71 µm. The particle size distribution of MPs showed fewer large particles than expected from a three-dimensional fractal fragmentation pattern, which was attributed to the higher mobility of small particles, especially fibres. The atmospheric deposition rate of MPs ranged from 5.6 to 78.6 MPs m-2 day-1, with the higher values observed in densely populated areas such as Barcelona and Madrid. Additionally, we detected natural polymers, mostly cellulosic fibres with evidence of industrial processing, with a deposition rate ranging from 6.4 to 58.6 particles m-2 day-1. There was a positive correlation was found between the population of the study area and the median of atmospheric MP deposition, supporting the hypothesis that urban areas act as sources of atmospheric MPs. Our study presents a systematic methodology for monitoring atmospheric MP deposition.

Recycled wastewater as a potential source of microplastics in irrigated soils from an arid-insular territory (Fuerteventura, Spain).

In this work, the occurrence of microplastics (MPs) in irrigation recycled wastewaters (RWWs) and a desalinated brackish water (DBW) from the arid territory of Fuerteventura (Canary Islands, Spain) was studied. Besides, the presence of MPs in two types of soils (sandy-loam and clay-loamy; with no mulch film or fertilization with sewage sludge applied) irrigated with both water qualities was addressed. Results showed the prevalence presence of cellulosic and polyester microfibers (between 84.4 and 100%) of blue and transparent colors (up to 55.6 and 33.3%, respectively), with an average length of 786.9 ± 812.1 µm in the water samples. DBW had the lowest MP concentration (2.0 ± 2.0 items·L-1) while RWW showed concentrations up to 40.0 ± 19.0 items·L-1. Similarities were also observed between the MPs types and sizes found in both soils top layer (0-5 cm), with an average concentration three times greater in soil irrigated with RWW than in soil under DBW irrigation (159 ± 338 vs. 46 ± 92 items·kg-1, respectively). In addition, no MPs were extracted from non-irrigated/non-cultivated soils, suggesting agricultural activities as the unique source of MPs in soils of this arid area. Results show that RWWs constitute a potential source of MPs in irrigated soils that should be considered among other pros and cons linked to the use of this water quality in agricultural arid lands.

Assessment of microplastic content in Diadema africanum sea urchin from Tenerife (Canary Islands, Spain).

Sea urchins are highly abundant in the marine ecosystem where they graze limiting algal biomass and also serving as food for other predators. In this work, the presence of microplastics in the digestive tracts and gonads of 33 Diadema africanum sea urchins collected at two sampling points in Tenerife (Canary Islands, Spain) was studied. After separation and digestion of the digestive tracts and the gonads, the visualization of the filtrates under the stereomicroscope revealed the presence of 320 items which were microfibers (97.5%), fragments (1.9%) and films (0.6%), mainly blue (43.3 and 47.0% in the two sampling points, Tajao and El Porís, respectively) and translucent white (32.5 and 39.5%, respectively). Statistical analysis revealed that there were no significative differences in the contents of gonads and digestive tracts between both sampling locations. Regarding microfibers lengths, significative differences were only observed between the two sampling points, not between tissues. µRaman analysis showed that they were mainly cellulosic (46.0%), polypropylene (24.3%) and polyethylene terephthalate (24.3%). This study confirms for the first time the presence of microplastics in sea urchins from the Macaronesian region and also from Spain.

Microplastic pollution in sublittoral coastal sediments of a North Atlantic island: The case of La Palma (Canary Islands, Spain).

In this work, the microplastic content of sediments collected in July 2020 between 5 and 7 m depth was studied in four locations of La Palma island (Canary Islands, Spain). At each sampling location, three samples were taken parallel to the shoreline. The microplastic content in each sampling corer was studied every 2.5 cm depth after digestion with a H2O2 solution followed by flotation in a saturated NaCl solution. Visualization of the final filtrates under a stereomicroscope revealed that all the sediment samples evaluated contained mostly microfibers (98.3%) which were mainly white/colorless (86.0%) and blue (9.8%), with an average length of 2423 ± 2235 (SD) mm and an average concentration of 2682 ± 827 items per kg of dry weight, being the total number of items found 1,019. Fourier Transform Infrared microscopy analysis of 13.9% (n = 139) of the microfibers also showed that they were mainly cellulosic (81.3%). No significant differences were found between the depths of the sediment. However, significant differences were found between the number of fibers from the sampling sites at the east and west of the island. Such variability could be driven by the winds and ocean mesoscale dynamics in the area. This study confirms the wide distribution of microfibers in sediments from an oceanic island like La Palma, providing their first report in marine sediments of the Canary Islands.

Plastitar: A new threat for coastal environments.

Oil residues have been frequently found on the coasts all over the world as a result of different accidental releases. Their partial evaporation and solidification onto the coastal rocks can produce the formation of a new solid structure forming an agglomerate with other materials, mainly microplastics (though wood, glass, sand and rocks were also found), yielding to a new plastic formation, name herein for the first time as "plastitar". These new formations have been found in several of the islands of the Canary Islands archipelago (Spain). Their study has shown that these new formations can be permanently attached to the rock, occupying even a 56% of the sampled area with an heterogeneous distribution. It was also observed that the studied plastitar was composed mainly of tar and polyethylene (90.6% of the studied particles) and polypropylene (9.4% of the studied particles) microplastics, primarily fragments (82.5%), pellets (15.7%) and lines (1.8%). The ever more frequent presence of plastics and, in particular, microplastics in coastal environments can lead to the common occurrence of these new plastic formations (probably present in other parts of the world), which long-term effects on the coasts should be further investigated.

Microplastics Determination in Gastrointestinal Tracts of European Sea Bass (Dicentrarchus labrax) and Gilt-Head Sea Bream (Sparus aurata) from Tenerife (Canary Islands, Spain).

Microplastic pollution has an extremely widespread distribution, to the extent that microplastics could be ingested by aquatic organisms, including species of commercial importance for fisheries and aquaculture. In this work, the anthropogenic particles content of the gastrointestinal tracts of 86 individuals of cultivated European sea bass (Dicentrarchus labrax, n = 45) and gilt-head sea bream (Sparus aurata, n = 41) from Tenerife (Canary Islands, Spain) was determined. Samples were bought at local markets and directly transported to the laboratory. After the dissection of the fishes and digestion of the gastrointestinal tracts in 10% KOH (w/v) at 60 °C for 24 h, the digests were filtered (50 µm stainless-steel mesh) and visualized under a stereomicroscope, finding that most of the items were colourless (47.7% for Dicentrarchus labrax and 60.9% for Sparus aurata) and blue (35.3% vs. 24.8%) microfibers, with an average length of 1957 ± 1699 µm and 1988 ± 1853 µm, respectively. Moreover, 15.3% of the microfibres were analysed by Fourier transform infrared spectroscopy, showing the prevalence of cellulosic fibres together with polyester, polyacrylonitrile, and poly(ether-urethane). This pattern (microplastics shapes, colours, sizes, and composition) clearly agrees with previous studies carried out in the Canary Islands region regarding the determination of microplastics in the marine environment.

Arenas Blancas (El Hierro island), a new hotspot of plastic debris in the Canary Islands (Spain).

The present work has studied the incidence and type of micro (1-5 mm), meso (5-25 mm) and macroplastics (>25 mm) that have reached Arenas Blancas beach, located in the north coast of El Hierro island, in the Canary Islands (Spain), from October 2019 to May 2020 (13 sampling dates with 3 sampling points each). Taking into consideration the three studied plastic debris fractions (macro, meso and microplastics), a total of 9206 items were found, which had a total weight of 1169.7 g and a concentration of 891.3 ± 91.5 items/m2 (118.3 ± 17.8 g/m2 and 2.3 ± 0.4 g/L). Regarding their colour, most of them were transparent/white/clear, especially in the microplastic fraction in which they accounted for a 68% of the total. Attenuated Total Reflectance Fourier Transform Infrared spectroscopy of meso and microplastic fractions indicated that most of the particles were either polypropylene and polyethylene followed by polystyrene in a much lower amount. In general, the total amount of plastic debris that arrives to the beach by the persistent oceanic current pattern linked to the easternmost branch of the North Atlantic Subtropical Gyre is comparable to those of the most contaminated beaches of the Canary Islands archipelago, suggesting that a new hotspot of plastic debris arrival has been found.

Engineered ACC deaminase-expressing free-living cells of Mesorhizobium loti show increased nodulation efficiency and competitiveness on Lotus spp.

Ethylene inhibits the establishment of symbiosis between rhizobia and legumes. Several rhizobia species express the enzyme ACC deaminase, which degrades the ethylene precursor 1-cyclopropane-1-carboxilate (ACC), leading to reductions in the amount of ethylene evolved by the plant. M. loti has a gene encoding ACC deaminase, but this gene is under the activity of the NifA-RpoN-dependent promoter; thus, it is only expressed inside the nodule. The M. loti structural gene ACC deaminase (acdS) was integrated into the M. loti chromosome under a constitutive promoter activity. The resulting strain induced the formation of a higher number of nodules and was more competitive than the wild-type strain on Lotus japonicus and L. tenuis. These results suggest that the introduction of the ACC deaminase activity within M. loti in a constitutive way could be a novel strategy to increase nodulation competitiveness of the bacteria, which could be useful for the forage inoculants industry.

Foot-and-Mouth Disease: Optimization, Reproducibility, and Scalability of High-Yield Production of Virus-Like Particles for a Next-Generation Vaccine.

Inactivated Foot-and-Mouth Disease (FMD) vaccine has proven to be effective in the control of the disease. However, its production has some disadvantages, including the costly biosafety facilities required for the production of huge amounts of growing live virus, the need of an exhaustive purification process to eliminate non-structural proteins of the virus in the final formulations in order to differentiate infected from vaccinated animals and variable local regulatory restrictions to produce and commercialize the vaccine. Thus, a novel vaccine against FMD that overcome these restrictions is desirable. Although many developments have been made in this regard, most of them failed in terms of efficacy or when considering their transferability to the industry. We have previously reported the use of transient gene expression in mammalian cells to produce FMD virus-like particles (VLPs) as a novel vaccine for FMD and demonstrated the immunogenicity of the recombinant structures in animal models. Here, we report the optimization of the production system by assaying different DNA:polyethylenimine concentrations, cell densities, and direct and indirect protocols of transfection. Also, we evaluated the reproducibility and scalability of the technology to produce high yields of recombinant VLPs in a cost-effective and scalable system compatible with industrial tech-transfer of an effective and safe vaccine.

Monitoring of meso and microplastic debris in Playa Grande beach (Tenerife, Canary Islands, Spain) during a moon cycle.

The occurrence and composition of meso (5-25 mm) and microplastics (1-5 mm) in Playa Grande beach (Tenerife, Canary Islands, Spain) was monitored during a complete moon cycle on the different moon phases between 17th June and 16thJuly 2019. A total of 10 points were sampled each day finding an average content of mesoplastics of 18 g/m2 (0.36 g/L) and of microplastics of 13 g/m2 (1277 items/m2 or 1.6 g/L). Polypropylene and polyethylene accounted for 19% and 76% of the total, respectively. Tar was also found in the 1-5 mm fraction (2% of the total). Among the particles found, 83% were fragments, 11% pellets, 4% fibres and 2% films. The obtained results revealed that microplastic presence could not be related in this case with the tides but with the orientation and strength/speed of the wind.

Characterization of the Mesorhizobium loti MAFF303099 type-three protein secretion system.

Type III secretion systems (T3SS) have been found in several species of rhizobia. Proteins (termed effectors) secreted by this system are involved in host-range determination and influence nodulation efficiency. Mesorhizobium loti MAFF303099 possesses a functional T3SS in its symbiotic island whose expression is induced by flavonoids. As in other rhizobia, conserved cis-elements (tts box) were found in the promoter regions of genes or operons encoding T3SS components. Using a bioinformatics approach, we searched for other tts-box-controlled genes, and confirmed this transcriptional regulation for some of them using lacZ fusions to the predicted promoter regions. Translational fusions to a reporter peptide were created to demonstrate T3SS-mediated secretion of two new MAFF303099 effectors. Finally, we showed that mutation of the M. loti MAFF303099 T3SS affects its competitiveness on Lotus glaber and investigated, at the molecular level, responses of the model legume L. japonicus to the T3SS.