Persistence of Escherichia coli in the microbiomes of red Romaine lettuce (Lactuca sativa cv. 'Outredgeous') and mizuna mustard (Brassica rapa var. japonica) - does seed sanitization matter?

BACKGROUND: Seed sanitization via chemical processes removes/reduces microbes from the external surfaces of the seed and thereby could have an impact on the plants' health or productivity. To determine the impact of seed sanitization on the plants' microbiome and pathogen persistence, sanitized and unsanitized seeds from two leafy green crops, red Romaine lettuce (Lactuca sativa cv. 'Outredgeous') and mizuna mustard (Brassica rapa var. japonica) were exposed to Escherichia coli and grown in controlled environment growth chambers simulating environmental conditions aboard the International Space Station. Plants were harvested at four intervals from 7 days post-germination to maturity. The bacterial communities of leaf and root were investigated using the 16S rRNA sequencing while quantitative polymerase chain reaction (qPCR) and heterotrophic plate counts were used to reveal the persistence of E. coli. RESULT: E. coli was detectable for longer periods of time in plants from sanitized versus unsanitized seeds and was identified in root tissue more frequently than in leaf tissue. 16S rRNA sequencing showed dynamic changes in the abundance of members of the phylum Proteobacteria, Firmicutes, and Bacteroidetes in leaf and root samples of both leafy crops. We observed minimal changes in the microbial diversity of lettuce or mizuna leaf tissue with time or between sanitized and unsanitized seeds. Beta-diversity showed that time had more of an influence on all samples versus the E. coli treatment. CONCLUSION: Our results indicated that the seed surface sanitization, a current requirement for sending seeds to space, could influence the microbiome. Insight into the changes in the crop microbiomes could lead to healthier plants and safer food supplementation.

Early detection of metastatic disease in asymptomatic breast cancer patients with whole-body imaging and defined tumour marker increase.

BACKGROUND: Follow-up care in breast cancer is still an issue of debate. Diagnostic methods are more sensitive, and more effective therapeutic options are now available. The risk of recurrence is not only influenced by tumour stage but also by the different molecular subtypes. This study was performed to evaluate the use of whole-body imaging combined with tumour marker monitoring for the early detection of asymptomatic metastatic breast cancer (MBC). METHODS: This analysis was performed as part of a follow-up study evaluating 813 patients with a median follow-up of 63 months. After primary therapy, all patients underwent tumour marker monitoring for CEA, CA 15-3 and CA 125 at 6-week intervals within an intensified diagnostic aftercare algorithm. A reproducible previously defined increase was considered as a strong indicator of MBC. From 2007 to 2010, 44 patients with tumour marker increase underwent whole-body magnetic resonance imaging and/or an FDG-PET/CT scan. Histological clarification and/or imaging follow-up were done. RESULTS: Metastases were detected in 65.9% (29/44) of patients, 13.6% (6/44) had secondary malignancies besides breast cancer and 20.5% (9/44) had no detectable malignancy. Limited disease was found in 24.1% (7/29) of patients. Median progression-free survival of MBC was 9.2 months and median overall survival was 41.1 months. The 3- and 5-year survival rates were 64.2% and 40.0%, respectively. CONCLUSIONS: A reproducible tumour marker increase followed by whole-body imaging is highly effective for early detection. By consequence, patients might benefit from earlier detection and improved therapeutic options with a prolonged survival.

Leaf yield and mineral content of mizuna in response to cut-and-come-again harvest, substrate particle size, and fertilizer formulation in a simulated spaceflight environment.

The Veggie plant-growth unit deployed onboard the International Space Station (ISS) grows leafy vegetables to supplement crew diets. "Cut-and-come-again" harvests are tested to maximize vegetative yield while minimizing crew time. Water, oxygen, and fertilizer delivery to roots of leafy greens growing in microgravity have become the center of attention for Veggie. Current Veggie technology wicks water into particulate root substrates incorporating controlled-release fertilizer (CRF). Mizuna mustard (Brassica rapa) was grown under ISS-like environments in ground-based Veggie-analogue units comparing crop response to combinations of two different substrate particle sizes, two different fertilizer formulations, and three leaf-harvest times from each plant. Fine-particle porous ceramic substrate (Profile©) was compared with a 40:60 mix of fine-particle porous ceramic Profile© + coarse porous ceramic Turface© substrate. Identical 18-6-8 (NPK) CRF formulations consisting of [50% fast-release (T70) + 50% intermediate-release (T100) prills] vs. [50% fast-release (T70) + 50% slow-release (T180) prills] were incorporated into each substrate, and leaf tissues were harvested from each treatment combination at 28, 48, and 56 days after sowing. The combination of T100 CRF in 100% Profile© substrate gave the highest fresh mass (FM) and leaf area (LA) across harvests, whereas T180 CRF in 40% Profile© gave the lowest. Dry-mass (DM) yields varied with effects on leaf area. Tissue nitrogen (N) and potassium (K) specific contents declined across harvests for all treatment combinations but tended to be highest for T100 CRF/100% Profile©, and lowest for T180 CRF/40% Profile©. These major macronutrients were taken up faster by roots growing in small particle-size substrate incorporating intermediate-rate CRF, but also were depleted faster from the same treatment combination, suggesting it may not continue to be the best combination for additional harvests. Micronutrients did not decline in tissue specific content across treatment combinations, but manganese (Mn) accumulated in leaf tissue across treatments and apparently comes mainly from the ceramic substrate, regardless of particle size. Substrate leachate analysis following final harvest indicated that pH remained in the range for nominal availability of mineral nutrients for root uptake, but electro-conductivity measurements suggested depletion of fertilizer salts from root zones, especially from the treatment combination supporting the highest yields and major macronutrient contents. Although 100% Profile© was the better growth substrate for mizuna in combination with intermediate-rate CRF and three cut-and-come-again harvests in ground-based studies, mixed-particle-size substrates may be a better choice for plant growth under microgravity conditions, where capillary forces predominant and tend to cause saturation of a fine medium with water. Since there were no statistically significant interactions between substrate and fertilizer in this study, our ground-based findings for CRF choice should translate to the best substrate choice for microgravity, but if NASA wants to consider additional cut-and-come-again harvests from the same mizuna plants, more complex CRF formulations likely will have to be investigated.

Synbiotic yogurt consumption by healthy adults and the elderly: the fate of bifidobacteria and LGG probiotic strain.

Synbiotic dietary supplements are used to modulate the intestinal microbiota in adults; however, studies regarding elderly people are limited. The aim of this work is to determine the effect of regular consumption of yogurt containing Lactobacillus rhamnosus GG (LGG) and the fructooligosaccharide (FOS) Actilight(®) both in healthy adults and the elderly suffering from constipation. Yogurt was administered daily for 4 weeks; faecal samples were collected before starting, after 4-week administration and a 15-day wash out period, and analyzed for LGG and bifidobacteria counts. Constipation status in the elderly was recorded by a standardized questionnaire. Bifidobacteria did not increase in either groups; at the end of the intervention trial, LGG was detected in significantly different amount in the faeces of the elderly and adults, being higher in adults (95% vs. 30%). An increased evacuation number in the elderly was evidenced, probably due to the presence of FOS in yogurt.

Comparison of two controlled-release fertilizer formulations for cut-and-come-again harvest yield and mineral content of Lactuca sativa L. cv. Outredgeous grown under International Space Station environmental conditions.

Red Romaine leaf lettuce (Lactuca sativa L. cv. Outredgeous) was grown in ground-based analogues of the Veggie plant-growth units used to grow salad vegetables for astronauts on the International Space Station (ISS). Plants were grown for 56 days with three "cut-and-come again" leaf harvests from the same plants. Six Biomass-Production-Systems-for-Education (BPSe) units were used to grow 'Outredgeous' ('OR') lettuce in a walk-in growth chamber under temperature, humidity, and LED-lighting conditions similar to those occurring in Veggie on ISS. Because of the ISS micro-gravity environment, both Veggie and ground-based BPSe units utilize one-way capillary wicking of water into an arcillite clay root substrate. In the present study, two different controlled-release fertilizer (CRF) formulations incorporated into the arcillite were compared for effects on 'OR' growth rate, overall yield, and mineral content of leaves harvested from the same plants 28, 48, and 56 days after planting. Both CRF treatments had a rapid-releasing T70 component that kept growth rate equivalent over the first two harvests. Growth rate for both CRF treatments increased from the first to the second harvest, but then declined from the second to the third harvest, more so for the slower-releasing T180 CRF than for the moderately-releasing T100 CRF. Tissue content of the macro-nutrients N, P, and K declined at each harvest for both CRFs, while content of the micro-nutrients B, Zn, and Mn increased. Although pH did not go out of the nominal range for availability of mineral nutrients to roots over the cropping cycle, and electrical-conductivity of rootzone salts was neither excessive nor depleted, tissue macronutrient depletion and micro-nutrient accumulation may have contributed to yield declines. Although either CRF formulation can support adequate yield of 'OR' lettuce over a 56-day period, the moderately-releasing T100 formulation tends to give slightly higher yield, especially during the last growth increment, and with non-deficient mineral content.

Response of Arabidopsis thaliana and Mizuna Mustard Seeds to Simulated Space Radiation Exposures.

One of the major concerns for long-term exploration missions beyond the Earth's magnetosphere is consequences from exposures to solar particle event (SPE) protons and galactic cosmic rays (GCR). For long-term crewed Lunar and Mars explorations, the production of fresh food in space will provide both nutritional supplements and psychological benefits to the astronauts. However, the effects of space radiation on plants and plant propagules have not been sufficiently investigated and characterized. In this study, we evaluated the effect of two different compositions of charged particles-simulated GCR, and simulated SPE protons on dry and hydrated seeds of the model plant Arabidopsis thaliana and the crop plant Mizuna mustard [Brassica rapa var. japonica]. Exposures to charged particles, simulated GCRs (up to 80 cGy) or SPEs (up to 200 cGy), were performed either acutely or at a low dose rate using the NASA Space Radiation Laboratory (NSRL) facility at Brookhaven National Lab (BNL). Control and irradiated seeds were planted in a solid phytogel and grown in a controlled environment. Five to seven days after planting, morphological parameters were measured to evaluate radiation-induced damage in the seedlings. After exposure to single types of charged particles, as well as to simulated GCR, the hydrated Arabidopsis seeds showed dose- and quality-dependent responses, with heavier ions causing more severe defects. Seeds exposed to simulated GCR (dry seeds) and SPE (hydrated seeds) had significant, although much less damage than seeds exposed to heavier and higher linear energy transfer (LET) particles. In general, the extent of damage depends on the seed type.

A prospective multicenter phase II study of oral and i.v. vinorelbine plus trastuzumab as first-line therapy in HER2-overexpressing metastatic breast cancer.

BACKGROUND: To evaluate the efficacy and safety of oral and i.v. vinorelbine plus trastuzumab as first-line regimen in a patient-convenient application for human epidermal growth factor receptor 2 (HER2)-overexpressing patients with metastatic breast cancer. PATIENTS AND METHODS: Forty-two women were enrolled in a multicenter study. The patients received i.v. vinorelbine at a dose of 25 mg/m(2) on day 1 followed by oral vinorelbine at a dose of 60 mg/m(2) on days 8 and 15 in a 3-week cycle. Standard dose trastuzumab was given at 3-week intervals. RESULTS: Complete response was observed in 7 patients (18.9%) and partial response in 19 patients (51.4%), for an overall response rate of 70.3% [95% confidence interval (CI) 53.0-84.1]. The disease control rate reached 91.9% (95% CI 78.1-98.3). The median time to progression was 9.3 months, while median overall survival reached 35.6 months. Hematological and non-hematological toxic effects were acceptable with grade 3-4 leukopenia of 14% and neutropenia of 38%; cardiac toxicity did not reach the level of clinical relevance. CONCLUSION: The combination of i.v. and oral vinorelbine plus trastuzumab demonstrates high activity and good tolerability in first-line treatment of HER2-overexpressing metastatic breast cancer. In addition, it offers convenience for the patients with only one i.v. treatment every 3 weeks.

Altered Activity of Lateral Orbitofrontal Cortex Neurons in Mice following Chronic Intermittent Ethanol Exposure.

The lateral orbitofrontal cortex (LOFC) is thought to encode information associated with consumption of rewarding substances and is essential for flexible decision-making. Indeed, firing patterns of LOFC neurons are modulated following changes in reward value associated with an action outcome relationship. Damage to the LOFC impairs behavioral flexibility in humans and is associated with suboptimal performance in reward devaluation protocols in rodents. As chronic intermittent ethanol (CIE) exposure also impairs OFC-dependent behaviors, we hypothesized that CIE exposure would alter LOFC neuronal activity during alcohol drinking, especially under conditions when the reward value of ethanol was modulated by aversive or appetitive tastants. To test this hypothesis, we monitored LOFC activity using GCaMP6f fiber photometry in mice receiving acute injections of ethanol and in those trained in operant ethanol self-administration. In naive mice, an acute injection of ethanol caused a dose-dependent decrease in the frequency but not amplitude of GCaMP6f transients. In operant studies, mice were trained on a fixed ratio one schedule of reinforcement and were then separated into CIE or Air groups. Following four cycles of CIE exposure, GCaMP6f activity was recorded during self-administration of alcohol, alcohol+quinine (aversive), or alcohol+sucrose (appetitive) solutions. LOFC neurons showed discrete patterns of activity surrounding lever presses and surrounding drinking bouts. Responding for and consumption of ethanol was greatly enhanced by CIE exposure, was aversion resistant, and was associated with signs of LOFC hyperexcitability. CIE-exposed mice also showed altered patterns of LOFC activity that varied with the ethanol solution consumed.

Neonicotinoids in the agroecosystem: In-field long-term assessment on honeybee colony strength and microbiome.

Bees can be severely affected by various plant protection products (PPP). Among these, neonicotinoid insecticides are of concern as they have been shown to be responsible for extensive honeybee colonies death when released into the environment. Also, sublethal neonicotinoid doses contaminating single honeybees and their colonies (e.g. through contaminated pollen) are responsible for honeybees physiological alterations with probable implication also on microbiome functionality. Honeybees show symbiotic interactions with specific gut bacteria that can enhance the adult host performances. Among the known mechanisms, the modulation of the immune system, the degradation of recalcitrant secondary plant metabolites, pollen digestion, and hormonal signaling, are the most important functional benefits for the host honeybee. To date, few research efforts have aimed at revealing the impact of PPP on the gut microbial community of managed and wild honeybees. The majority of the existing literature relays on cage or semifield tests of short duration for research investigating neonicotinoids-gut microbiome interactions. This research wanted to unravel the impact of two neonicotinoids (i.e. imidacloprid and thiacloprid) in natural field conditions up to 5 weeks of exposure. A long-term impact of neonicotinoids on gut microbial community of honeybees was observed. The alterations affected several microbial genera and species such as Frischella spp., lactobacilli and bifidobacteria, whose shifting is implicated in intestinal dysbiosis. Long-term impact leading to dysbiosis was detected in case of exposure to imidacloprid, whereas thiacloprid exposure stimulated temporary dysbiosis. Moreover, the microbial diversity was significantly reduced in neonicotinoid-treated groups. Overall, the reported results support a compromised functionality of the gut microbial community, that might reflect a lower efficiency in the ecosystemic functionality of honeybees.

Evaluating the lettuce metatranscriptome with MinION sequencing for future spaceflight food production applications.

Healthy plants are vital for successful, long-duration missions in space, as they provide the crew with life support, food production, and psychological benefits. The microorganisms that associate with plant tissues play a critical role in improving plant health and production. To that end, we developed a methodology to investigate the transcriptional activities of the microbiome of red romaine lettuce, a key salad crop that was grown under International Space Station (ISS)-like conditions. Microbial transcripts enriched from host-microbe total RNA were sequenced using the Oxford Nanopore MinION sequencing platform. Results show that this enrichment approach was highly reproducible and could be an effective approach for the on-site detection of microbial transcriptional activity. Our results demonstrate the feasibility of using metatranscriptomics of enriched microbial RNA as a potential method for on-site monitoring of the transcriptional activity of crop microbiomes, thereby helping to facilitate and maintain plant health for on-orbit space food production.

Fusarium oxysporum as an Opportunistic Fungal Pathogen on Zinnia hybrida Plants Grown on board the International Space Station.

A plant production system called Veggie was launched to the International Space Station (ISS) in 2014. In late 2015, during the growth of Zinnia hybrida cv. 'Profusion' in the Veggie hardware, plants developed chlorosis, leaf curling, fungal growth that damaged leaves and stems, and eventually necrosis. The development of symptoms was correlated to reduced air flow leading to a significant buildup of water enveloping the leaves and stems in microgravity. Symptomatic tissues were returned to Earth on 18 May 2016 and were immediately processed to determine the primary causal agent of the disease. The presumptive pathogen was identified as Fusarium oxysporum by morphological features of microconidia and conidiophores on symptomatic tissues; that is, by epifluorescent microscopy (EFM), scanning electron microscopy (SEM), metabolic microarrays, and ITS sequencing. Both EFM and SEM imaging of infected tissues showed that germinating conidia were capable of stomatal penetration and thus acted as the primary method for infecting host tissues. A series of ground-based pathogenicity assays were conducted with healthy Z. hybrida plants that were exposed to reduced-airflow and high-water stress (i.e., encased in sealed bags) or were kept in an unstressed configuration. Koch's postulates were successfully completed with Z. hybrida plants in the lab, but symptoms only matched ISS-flown symptomatic tissues when the plants were stressed with high-water exposure. Unstressed plants grown under similar lab conditions failed to develop the symptoms observed with plants on board the ISS. The overall results of the pathogenicity tests imply that F. oxysporum acted as an opportunistic pathogen on severely high-water stressed plants. The source of the opportunistic pathogen is not known, but virulent strains of F. oxysporum were not recovered from unused materials in the Veggie plant pillow growth units assayed after the flight.

Spatial Characterization of Microbial Communities on Multi-Species Leafy Greens Grown Simultaneously in the Vegetable Production Systems on the International Space Station.

The establishment of steady-state continuous crop production during long-term deep space missions is critical for providing consistent nutritional and psychological benefits for the crew, potentially improving their health and performance. Three technology demonstrations were completed achieving simultaneous multi-species plant growth and the concurrent use of two Veggie units on the International Space Station (ISS). Microbiological characterization using molecular and culture-based methods was performed on leaves and roots from two harvests of three leafy greens, red romaine lettuce (Lactuca sativa cv. 'Outredgeous'); mizuna mustard, (Brassica rapa var japonica); and green leaf lettuce, (Lactuca sativa cv. Waldmann's) and associated rooting pillow components and Veggie chamber surfaces. Culture based enumeration and pathogen screening indicated the leafy greens were safe for consumption. Surface samples of the Veggie facility and plant pillows revealed low counts of bacteria and fungi and are commonly isolated on ISS. Community analysis was completed with 16S rRNA amplicon sequencing. Comparisons between pillow components, and plant tissue types from VEG-03D, E, and F revealed higher diversity in roots and rooting substrate than the leaves and wick. This work provides valuable information for food production-related research on the ISS and the impact of the plant microbiome on this unique closed environment.

Growth and photosynthetic responses of Chinese cabbage (Brassica rapa L. cv. Tokyo Bekana) to continuously elevated carbon dioxide in a simulated Space Station "Veggie" crop-production environment.

Among candidate leafy vegetable species initially considered for astronauts to pick and eat from the Veggie plant-growth unit on the International Space Station (ISS), Chinese cabbage (Brassica rapa L. cv. Tokyo Bekana) ranked high in ground-based screening studies. However, subsequent attempts to optimize growth within rigorous ISS-like growth environments on the ground were frustrated by development of leaf chlorosis, necrosis, and uneven growth. 'Tokyo Bekana' ('TB') grown on ISS during the VEG-03B and C flights developed similar stress symptoms. After lengthy troubleshooting efforts to identify causes of sub-par growth in highly controlled environments, the super-elevated CO2 concentrations that plants on ISS are exposed to continuously (average of 2,800 µmol/mol) emerged as a candidate environmental condition responsible for the observed plant-stress symptoms. Subsequent ground-based studies found continuous exposure to ISS levels of CO2 under Veggie environmental and cultural conditions to significantly inhibit growth of 'TB' compared to near-Earth-normal CO2 controls. The present study investigated growth and gas-exchange responses of 'TB' to sub-ISS but still elevated CO2 levels (900 or 1,350 µmol/mol) in combination with other potential stressors related to ISS/Veggie compared to 450 µmol/mol CO2 controls. Shoot dry mass of plants grown at 450 µmol•mol-1 CO2 for 28 days was 96% and 80% higher than that of plants grown at 900 µmol•mol-1 CO2 and 1,350 µmol•mol-1 CO2, respectively. Leaf number and leaf area of controls were significantly higher than those of plants grown at 1,350 µmol•mol-1 CO2. Photosynthetic rate measured using a leaf cuvette was significantly lower for plants grown at 900 µmol•mol-1 CO2 than for controls. The ratio of leaf internal CO2 concentration (Ci) to cuvette ambient CO2 concentration (Ca) was significantly lower for plants grown at 450 µmol•mol-1 CO2 than for plants grown at elevated CO2. Thus, continuously elevated CO2 in combination with a Veggie cultivation system decreased growth, leaf area, and photosynthetic efficiency of Chinese cabbage 'Tokyo Bekana'. The results of this study suggest that 'Tokyo Bekana' is very sensitive to continuously elevated CO2 in such a growth environment, and indicate the need for improved environmental control of CO2 and possibly root-zone factors for successful crop production in the ISS spaceflight environment. Differential sensitivity of other salad crops to an ISS/Veggie growth environment also is possible, so it is important to mimic controllable ISS-like environmental conditions as precisely as possible during ground-based screening.

Microbiological and Nutritional Analysis of Lettuce Crops Grown on the International Space Station.

The ability to grow safe, fresh food to supplement packaged foods of astronauts in space has been an important goal for NASA. Food crops grown in space experience different environmental conditions than plants grown on Earth (e.g., reduced gravity, elevated radiation levels). To study the effects of space conditions, red romaine lettuce, Lactuca sativa cv 'Outredgeous,' plants were grown in Veggie plant growth chambers on the International Space Station (ISS) and compared with ground-grown plants. Multiple plantings were grown on ISS and harvested using either a single, final harvest, or sequential harvests in which several mature leaves were removed from the plants at weekly intervals. Ground controls were grown simultaneously with a 24-72 h delay using ISS environmental data. Food safety of the plants was determined by heterotrophic plate counts for bacteria and fungi, as well as isolate identification using samples taken from the leaves and roots. Molecular characterization was conducted using Next Generation Sequencing (NGS) to provide taxonomic composition and phylogenetic structure of the community. Leaves were also analyzed for elemental composition, as well as levels of phenolics, anthocyanins, and Oxygen Radical Absorbance Capacity (ORAC). Comparison of flight and ground tissues showed some differences in total counts for bacteria and yeast/molds (2.14 - 4.86 log10 CFU/g), while screening for select human pathogens yielded negative results. Bacterial and fungal isolate identification and community characterization indicated variation in the diversity of genera between leaf and root tissue with diversity being higher in root tissue, and included differences in the dominant genera. The only difference between ground and flight experiments was seen in the third experiment, VEG-03A, with significant differences in the genera from leaf tissue. Flight and ground tissue showed differences in Fe, K, Na, P, S, and Zn content and total phenolic levels, but no differences in anthocyanin and ORAC levels. This study indicated that leafy vegetable crops can produce safe, edible, fresh food to supplement to the astronauts' diet, and provide baseline data for continual operation of the Veggie plant growth units on ISS.

Selection and characterization of aerobic bacteria capable of degrading commercial mixtures of low-ethoxylated nonylphenols.

AIMS: Isolation and characterization of new bacterial strains capable of degrading nonylphenol ethoxylates (NPnEO) with a low ethoxylation degree, which are particularly recalcitrant to biodegradation. METHODS AND RESULTS: Seven aerobic bacterial strains were isolated from activated sludges derived from an Italian plant receiving NPnEO-contaminated wastewaters after enrichment with a low-ethoxylated NPnEO mixture. On the basis of 16S rDNA sequence, the strains were positioned into five genera: Ochrobactrum, Castellaniella, Variovorax, Pseudomonas and Psychrobacter. Their degradation capabilities have been evaluated on two commercial mixtures, i.e. Igepal CO-210 and Igepal CO-520, the former rich in low ethoxylated congeners and the latter containing a broader spectrum of NPnEO, and on 4-n-nonylphenol (NP). The strains degraded Igepal CO-210, Igepal CO-520 and 4-n-NP all applied at the initial concentration of 100 mg l(-1), by 35-75%, 35-90% and 15-25%, respectively, after 25 days of incubation. CONCLUSIONS: Some of the isolated strains, in particular the Pseudomonas strains BCb12/1 and BCb12/3, showed interesting degradation capabilities towards low ethoxylated NPnEO congeners maintaining high cell vitality. SIGNIFICANCE AND IMPACT OF THE STUDY: Increased knowledge of bacteria involved in NPnEO degradation and the possibility of using the isolated strains in tailored process for a tertiary biological treatment of effluents of wastewater treatment plants.

Impact of beneficial bacteria supplementation on the gut microbiota, colony development and productivity of Apis mellifera L.

Honey bees are important pollinators of several crops and ecosystems, having a great ecological and economic value. In Europe, the restricted use of chemicals and therapeutic agents in the beekeeping sector has stimulated the search for natural alternatives with a special focus on gut symbionts. The modulation of the gut microbiota has been recognised as a practical and successful approach in the entomological field for the management of insect-related problems. To date, only a few studies have investigated the effect of bacterial supplementation on the health status of colonies, colony productivity and gut symbionts. To this purpose, a preparation of sugar syrup containing bifidobacteria and lactobacilli isolated from bee gut was sprayed on the frames of an apiary located in open field once a week for four weeks. Treated and control hives were monitored for two months for brood extension, honey and pollen harvest. The presence of beneficial gut microorganisms within bee gut was investigated with denaturing gradient gel electrophoresis and next generation sequencing. The administered bacteria led to a significant increase of brood population (46.2%), pollen (53.4%) and harvestable honey in honey supers (59.21%). Analysis of the gut microbiota on the new generation of bees in treated hives showed an increase in relative abundance of Acetobacteraceae and Bifidobacterium spp., which are known to be involved in bee nutrition and protection.

A phase II, multicentre trial of decitabine in higher-risk chronic myelomonocytic leukemia.

Chronic myelomonocytic leukemia (CMML) is a complex clonal hematological disorder classified among myelodysplastic (MDS)/myeloproliferative neoplasms. Prognosis is poor and there is a lack of effective treatments. The hypomethylating agent decitabine has shown activity against MDS and elderly acute myeloid leukemia, but there is little data focusing specifically on its efficacy in CMML. In this prospective, phase 2 Italian study, CMML patients received intravenous decitabine 20 mg/m2 per day on Days 1-5 of a 28-day treatment cycle. Response was evaluated after four and six cycles; patients responding at the end of six cycles could continue treatment with decitabine. Forty-three patients were enrolled; >50% were high-risk according to four CMML-specific scoring systems. In the intent-to-treat population (n=42), the overall response rate after six cycles was 47.6%, with seven complete responses (16.6%), eight marrow responses (19%), one partial response (2.4%) and four hematological improvements (9.5%). After a median follow-up of 51.5 months (range: 44.4-57.2), median overall survival was 17 months, with responders having a significantly longer survival than non-responders (P=0.02). Grade 3/4 anemia, neutropenia and thrombocytopenia occurred in 28.6%, 50% and 38% of patients, respectively. Decitabine appears to be an effective and well-tolerated treatment for patients with high-risk CMML.

The basal to total insulin ratio in outpatients with diabetes on basal-bolus regimen.

OBJECTIVE: To evaluate the basal/total ratio of daily insulin dose (b/T) in outpatients with diabetes type 1 (DM1) and type 2 (DM2) on basal-bolus regimen, by investigating whether there is a relationship with HbA1c and episodes of hypoglycemia. METHODS: Multicentric, observational, cross-sectional study in Italy. Adult DM1 (n = 476) and DM2 (n = 541) outpatients, with eGFR >30 mL/min/1.73 m2, on a basal-bolus regimen for at least six months, were recruited from 31 Italian Diabetes services between March and September 2016. Clinicaltrials.govID: NCT03489031. RESULTS: Total daily insulin dose was significantly higher in DM2 patients (52.3 ± 22.5 vs. 46 ± 20.9 U/day), but this difference disappeared when insulin doses were normalized for body weight. The b/T ratio was lower than 0.50 in both groups: 0.46 ± 0.14 in DM1 and 0.43 ± 0.15 in DM2 patients (p = 0.0011). The b/T was significantly higher in the patients taking metformin in both groups, and significantly different according to the type of basal insulin (Degludec, 0.48 in DM1 and 0.44 in DM2; Glargine, 0.44 in DM1 and 0.43 in DM2; Detemir, 0.45 in DM1 and 0.39 in DM2). The b/T ratio was not correlated in either group to HbA1c or incidence of hypoglycemia (<40 mg/dL, or requiring caregiver intervention, in the last three months). In the multivariate analysis, metformin use and age were independent predictors of the b/T ratio in both DM1 and DM2 patients, while the type of basal insulin was an independent predictor only in DM1. CONCLUSION: The b/T ratio was independent of glycemic control and incidence of hypoglycemia.

Correction to: The basal to total insulin ratio in outpatients with diabetes on basal-bolus regimen.

[This corrects the article DOI: 10.1007/s40200-018-0358-2.].

Effect of dietary supplementation of Bifidobacterium and Lactobacillus strains in Apis mellifera L. against Nosema ceranae.

Nosema ceranae is a widespread microsporidium of European honeybee Apis mellifera L. affecting bee health. The ban of Fumagillin-B (dicyclohexylammonium salt) in the European Union has driven the search for sustainable strategies to prevent and control the infection. The gut microbial symbionts, associated to the intestinal system of vertebrates and invertebrates and its impact on host health, are receiving increasing attention. In particular, bifidobacteria and lactobacilli, which are normal inhabitants of the digestive system of bees, are known to protect their hosts via antimicrobial metabolites, immunomodulation and competition. In this work, the dietary supplementation of gut bacteria was evaluated under laboratory conditions in bees artificially infected with the parasite and bees not artificially infected but evidencing a low natural infection. Supplemented bacteria were selected among bifidobacteria, previously isolated, and lactobacilli, isolated in this work from healthy honeybee gut. Four treatments were compared: bees fed with sugar syrup (CTR); bees fed with sugar syrup containing bifidobacteria and lactobacilli (PRO); bees infected with N. ceranae spores and fed with sugar syrup (NOS); bees infected with N. ceranae and fed with sugar syrup containing bifidobacteria and lactobacilli (NP). The sugar syrup, with or without microorganisms, was administered to bees from the first day of life for 13 days. N. ceranae infection was carried out individually on anesthetised 5-day-old bees. Eight days after infection, a significant (P<0.05) lower level of N. ceranae was detected by real-time PCR in both NP and PRO group, showing a positive effect of supplemented microorganisms in controlling the infection. These results represent a first attempt of application of bifidobacteria and lactobacilli against N. ceranae in honeybees.