We aimed to determine the effectiveness of glycaemic monitoring and control in the inpatient setting of a tertiary New Zealand hospital, and whether suboptimal control and monitoring may be associated with adverse outcomes in both Māori and Non-Māori diabetes patients.Clinical records including all glucose levels  from inpatients ≥ 15 years old with diabetes who were admitted to Waikato Hospital for > 24 hours between 1st July 2017 to 30th June 2018 were extracted electronically from the hospital database, and the data retrospectively examined .Overall 80.8% of diabetes inpatients had their blood glucose levels monitored. Patients experiencing ≥ 1 episode of hypoglycaemia were 1.90 times (CI: 1.37-2.64) and 1.94 times (CI: 1.51-2.49) more likely to die within 60 days and one year respectively, with an increased length of hospital stay by a mean of 3.13 weeks (CI: 2.55-3.85). Māori patients were more likely to experience ≥ 1 episode of hypoglycaemia (OR: 1.46), with a higher one-year mortality (p<0.001) as well as higher readmission rates at 30,60, 90 and 365 days than non-Māori. Blood glucose checks at least once every 24 hours were associated with shorter hospital stays (0.36 weeks) and a lower one-year mortality (Adjusted odds ratios (OR): 0.77, CI: 0.64-0.91).At least one episode of inpatient hypoglycaemia was associated with a statistically significant increase in 60-day and one-year mortality as well as notably longer hospital stays, with more frequent hypoglycaemia occurring in Māori patients. Significant hyperglycaemia was associated with an increased one-year mortality, higher readmission rates within one year and longer hospital stays. The estWhilst very tight glycaemic control may increasemortality in the critically ill , the ben2[1 We aimed to assess the frequency of glucose monitoring and control in such a setting as well as whether glycaemic abnormalities were associated with increased readmission rates and mortality.Currently there is a scarcity of published data relating to New Zealand inpatient glycaemic monitoring and control necessitating this retrospective review undertaken at a 673 bed New Zealand urban tertiary care hospital, with both an urban and rural catchment area of 25,598km2. In the 2. There wMāori have inequitable health outcomes in New Zealand, living seven years shorter than non-Māori, with reduced access to health services and higher rates of disability compared with non-Māori . We ther2st July 2017 to 30th June 2018 with an International Classification of Diseases (ICD) code of either type 1 or type 2 diabetes. Pregnant patients were excluded from the study due to different glycaemic recommendations [st March 2017 to 30th June 2018.Electronic data were obtained for all patients ≥ 15 years of age who were admitted to Waikato Hospital for > 24 hours between 1ndations . PatientData collected for analysis comprised: number of admissions in patients with diabetes irrespective of the cause of admission; number of patients; patient demographics including age, gender, ethnicity and socioeconomic status; frequency of capillary blood glucose and ketone monitoring; number and severity of hyperglycaemic and hypoglycaemic events; length of stay; critical care admissions. For the purposes of analysis, the following glycaemic ranges were used: Hypoglycaemia ≤ 3.9 mmol/L; target range blood glucose 4.0-10.0 mmol/L; mild hyperglycaemia 10.1-13.9 mmol/L; significant hyperglycaemia ≥ 14 mmol/L [Characteristics and outcomes of patient admissions were compared between those having at least one episode of hypoglycaemia (≤ 3.9 mmol/L), hyperglycaemia (> 10 mmol/L) and/or significant hyperglycaemia (≥ 14 mmol/L) to those without these events; by using Chi-squared tests for categorical variables and Student’s t-tests for continuous variables. The data was analysed to provide information regarding outcomes in relation to glycaemic parameters, length of stay, mortality, readmissions, critical care admissions, socioeconomic status and ethnicity. Ethnicity statistics were prioritised for Māori as this was an ethnicity of interest in this study. The New Zealand Deprivation 2013 (NZDep2013) index score was used as a marker of socioeconomic status, with higher scores indicating greater levels of social deprivation .Logistic regressions were used to examine the association between outcomes  and suboptimal monitoring of glucose levels and glycaemic control  adjusting for type of diabetes, ethnicity, and age. We also ran Ordinary Least Squares (OLS) regressions to investigate the impact of suboptimal monitoring of glucose levels and glycaemic control on the length of stay, adjusting for type of diabetes, ethnicity, and age. Logistic regressions were used to assess the ethnic differences in the monitoring of glucose levels and glycaemic control adjusting for age and type of diabetes. Data analyses in 3Over the selected time period a total cohort of 4,901 admissions in 3,380 diabetes patients was assessed, of which 80.8% (n=4901) had their blood glucose levels monitored . Individual patients were admitted a mean of 1.6 (+/-1.1) times during the 16-month study period, and the demographic and clinical characteristics of this group are shown in The mean age of participants was 67 years and admitted patients with at least one episode of hypoglycaemia, hyperglycaemia or significant hyperglycaemia tended to be younger . The mean length of stay was 1.3 weeks. Approximately 9.5% of admitted patients died within 60 days of admission and 22.6% of admitted patients died within one year. Overall, the mean number of glucose measurements taken over a 24-hour period was 0.7 (+/-0.8). One quarter  of admissions had one or more glucose level recorded for each 24-hour period of their hospital stay, though only 0.8% of admissions had blood glucose levels recorded at least four times .Overall, 61% of blood glucose measurements were in target range . A totalThere was a statistically significant association noted between hypoglycaemia and mortality as well as length of stay . ApproxiMāori patients accounted for 33.6% of total admissions. Māori were statistically significantly more likely than non-Māori to experience at least one episode of hypoglycaemia  and less likely to experience hyperglycaemia  or significant hyperglycaemia  after adjusting for age and type of diabetes. Māori patients with diabetes also had a statistically significant higher one-year mortality as well as higher readmission rates at 30, 60, 90 and 365 days than nonMāori patients .4The frequency of inpatient glycaemic monitoring at our tertiary centre is clearly suboptimal and far from meeting the recommended standards. Furthermore, a substantial number of the glucose levels recorded fall outside the recommended target range for in patients with diabetes. At the time of the study, Waikato hospital did not have a standardised tool to explicitly order point of care glucose and ketone testing leaving some ambiguity to the monitoring and frequency of testing. This reason along with clinical inertia may go some way to explaining why 19.2% of the admission cohort did not have any glucose monitoring, which is concerningly high and puts patients at risk of late detection of hypoglycaemia, hyperosmolar hyperglycaemic state and diabetic ketoacidosis. Other explanations postulated for this include lack of awareness of medical and nursing staff of the importance of frequent glycaemic monitoring, heavy nursing workloads as well as patients and nursing staff relying on the patient’s own glucometers while in hospital. A tool has subsequently been implemented at Waikato hospital allowing standardisation of inpatient diabetes care which should ameliorate the problem of poor glycaemic monitoring and control.Hyperglycaemia intensifies the body’s pro-inflammatory responses, potentiates oxidative injury and is pro-thrombotic thereby contributing to poorer outcomes in life threatening conditions including acute myocardial infarction, stroke and congestive cardiac failure . There iWe speculate reasons that Māori patients had more hypoglycaemia than non-Māori patients may include overly aggressive treatment of abnormal glucose measurements in hospital as well as recurrent covert non-adherence to diabetes treatment in the community leading to poorer glycaemic control resulting in repeated up-titration of insulin and diabetes medications, with subsequent hypoglycaemia when all these medications are given at once in hospital. Māori have substandard health outcomes and an increased risk of complications from diabetes in New Zealand . This stFurther, approximately two fifths of patients with diabetes did not have a HbA1c measurement from the preceding three months. The reasons for this are likely multifactorial including clinical inertia, measurement out of area, social deprivation as well as poor patient access to general practice and laboratory sites. Nevertheless, a hospital admission should have provided the opportunity for HbA1c testing with remediable action taken if suboptimal glycaemic control was identified.It is recommended that type 1 diabetes patients have a blood ketone level checked if the blood glucose is persistently over 15 mmol/L or if unwell . CheckinThere were several limitations identified in the study. Insulin and diabetes medication use in individual patients was not explored. Venous blood gas (VBG) analyses were not included in the study. VBG testing is an essential component of a diabetic ketoacidosis diagnosis and monitoring plan as well as being frequently used by intensive care units. However, there were only a small number of ICU and HDU patients involved in the study suggesting that inclusion of VBG glucose levels would not have significantly altered our results. Only electronic data was used but the laboratory ensured all point of care testing data was regularly uploaded from every ward during the study period. Those admitted for less than 24 hours were excluded from the study, but we hypothesise that rates of glucose and ketone monitoring may have been lower in this group due to milder illness.In the era of rapidly advancing diabetes technologies, it is more commonplace for diabetes patients to be utilising continuous glucose monitoring and flash glucose monitoring. Whilst these systems have been shown to improve glycaemic control , they haThis study demonstrates the need to prioritise glycaemic monitoring and control of inpatients with diabetes in order to enable more rapid recovery from acute illness, reduce morbidity, and decrease the length of hospital stays and costs. Previous studies have demonstrated that healthcare workers including nursing staff, pharmacists and junior doctors have low confidence when managing diabetes . With thStrategies focusing on the system of diabetes management as well as patient-mediated quality improvement factors are vital . CreatinIn conclusion, both significant hyperglycaemia and hypoglycaemia were associated with increased morbidity as well as longer hospital stays emphasising the importance of maintaining good glycaemic monitoring and control in hospitalised diabetes patients. Inpatient hypoglycaemia was associated with increased mortality rates and was more common in Māori patients with diabetes. At the time of the study, both monitoring and glycaemic control were suboptimal, with strategies required to improve this. However, given the ever-growing use of diabetes technologies and resources available to aid in diabetes management, a significant improvement in these results is expected in the near future.


Modern aquaculture is moving towards sustainable aquafeed formulations and farming techniques that allow the application of the circular economy concept and the reduction of water consumption. In this regard, the present study is focused on the use of black soldier fly full-fat prepupae meal (HPM) enriched with spirulina as a more sustainable aquafeed ingredient in diets intended for European seabass reared in aquaponic systems. The results obtained from the present study allowed us to demonstrate the suitability of aquaponic systems for the rearing of euryhaline finfish species and the beneficial properties of spirulina-enriched HPM on fish growth, welfare, and quality features.Hermetia illucens) prepupae meal  were tested on European seabass (Dicentrarchus labrax) juveniles during a 90-day feeding trial performed in aquaponic systems. The experimental diets ensured 100% survival and proper zootechnical performance. No behavioral alterations were evidenced in fish. Histological and molecular analyses did not reveal structural alterations and signs of inflammation at the intestinal level, highlighting the beneficial role on gut health of bioactive molecules typical of HPM or derived from the enriching procedure of insects’ growth substrate with spirulina. Considering the quality traits, the tested experimental diets did not negatively alter the fillet’s fatty acid profile and did not compromise the fillet’s physical features. In addition, the results highlighted a possible role of spirulina-enriched HPM in preserving the fillet from lipid oxidation. Taken together, these results corroborate the use of sustainable ingredients (spirulina-enriched HPM) in aquaponic systems for euryhaline fish rearing.Within the modern aquaculture goals, the present study aimed to couple sustainable aquafeed formulation and culturing systems. Two experimental diets characterized by 3 and 20% of fish meal replacement with full-fat spirulina-enriched black soldier fly ( The human population is expected to reach 8.5 billion by 2030 and 9.7 billion by 2050 , raisingRegarding aquafeed production, several ingredients have been investigated as promising alternatives to replace conventional and less sustainable marine-derived ingredients, such as fish meal (FM) and oil , includiHermetia illucens), have been approved when reared using feed-grade substrates [Among the alternatives, insects have recently gained increasing attention due to their many beneficial properties . Insectsbstrates .From a nutritional standpoint, the high protein content (around 60% or up to 65.5% in defatted larvae), well-balanced essential amino acid profile similar to FM, and the vitamin and mineral content makes the black soldier fly prepupae meal (HPM) an apparent suitable option for inclusion as ingredient in aquafeed formulations ,24. FurtIn this regard, black soldier fly larvae can modulate their fatty acid profile according to the composition of their feeding substrate ,32. In tArthrospira platensis), the most commercially produced species of cyanobacteria among phytoplankton, contains PUFA and numerous bioactive compounds comprising phycobiliproteins, carotenoids, phycocyanin, polysaccharides, and a number of vitamins, which have the ability to positively affect fish health and quality [Oncorhynchus mykiss) and giant freshwater prawn (Macrobrachium rosenbergii) culture, leading to positive physiological responses in both these species [Spirulina , an economically relevant marine species in the European Union aquaculture [In an ever-growing aquaculture, aquafeed formulations, together with water consumption, represent the key factors challenging the long-term sustainability of this sector. For this reason, this highly productive sector has to move towards more sustainable practices, placing emphasis on the circular economy concept ,5. Accoraculture . The triaculture . At the All protocols and procedures for fish manipulation were performed according to the guidelines established by the Council of the European Communities Directive (86/609/EEC and 2010/63/EU) for animal welfare and approved by the Animal Ethics Committee of Marche Polytechnic University (n. 01/2022-UT of 17 January 2022). Optimal rearing conditions were ensured, and animal suffering was minimized using the MS222 anesthetic .w/w) of the dried biomass from Arthrospira platensis was included, reaching a final moisture of ~70% by adding distilled water. The black soldier fly larvae  were reared on the feeding substrate placed in a climatic chamber set at 27 ± 1 °C and 65 ± 5% of relative humidity. Larvae were reared at a 0.3/cm2 density with a feeding rate of 100 mg/day per larva [Insects’ rearing was conducted following the procedures reported in Ratti et al. . Brieflyer larva ,43, ensuThree experimental diets were formulated to cover the nutritional requirements of European seabass . SpecifiThe proximate composition of the experimental diets was analyzed by following AOAC (2016) procedures , while tFor the fatty acid profile, a base-catalyzed transesterification of 4 mg total lipid in methyl esters (FAME)  was used2) of 0.999 were obtained in all cases.For the α-tocopherol and carotenoid determination, the protocols described by Ratti et al.  and Nart2, one day before the introduction of fish in the aquaponic systems. Each hydroponic unit was equipped with expanded clay to ensure physical support for plant growth and both mechanical and biological filtration for fish tanks.The feeding trial was conducted at Mj Energy srl Società Agricola . Two hundred and seventy European seabass juveniles, provided by Panittica Pugliese , were acclimated to the experimental conditions for two weeks in a single 1000 L tank equipped with mechanical, biological, and UV filtration . Water parameters were as follows: temperature, 17.0 ± 0.5 °C; salinity 30 ± 0.5‰; ammonia and nitrite < 0.05 mg/L; nitrate < 10 mg/L. During the acclimation period, salinity was gradually decreased from 30 to 5‰ (2‰ daily by adding freshwater) to match the salinity of aquaponic systems. At the end of the acclimation period, fish  were randomly divided into nine Media-Based Aquaponic Systems (600 L fish tank and 120 L hydroponic unit), with three systems per experimental group (30 specimens per tank). Fish were subjected to a natural photoperiod (13L/11D) and water in the aquaponic systems was maintained at a constant temperature (17.0 ± 0.2 °C) by TK500 chillers  and at 5 ± 0.5‰ of salinity. A 1900 L/h pump  regulated the water flow, completing 3 water renewals each hour. From each fish tank, water was pumped to the corresponding hydroponic unit, going back through a siphon in which synthetic foam was added to provide extra mechanical filtration. In each hydroponic unit, 100 saffron (Crocus sativus) seedlings were planted, at a density of 65 plants/mWater samples were collected once a week to monitor ammonia, nitrite, nitrate, and phosphate concentrations . The duration of the feeding trial was 90 days, during which fish almost triplicated their weight. Fish were hand-fed the experimental diets at 2% body weight, divided into one ration in the morning and one in the afternoon. The daily feed was adjusted every two weeks by weighing a representative number of specimens for each tank. Fish from the HPM0 group were fed the control diet, while fish from the HPM3 and HPM20 groups were fed diets characterized by 3 or 20% of FM replacement with HPM, respectively. The feed particle size was 3 mm. At the end of the trial, all the fish from each tank were euthanized with a lethal dose of anesthetic , then counted and individually weighted. The whole fish and their fillets, as well as samples from the liver and distal intestine, were collected and properly stored for further analyses.For each tank, survival rate (SR), relative growth rate (RGR), specific growth rate (SGR), and feed conversion ratio (FCR) were calculated as follows:The behavioral test was performed one week before the end of the trial. Fish exploration tendency in a novel environment was individually assayed using an open-field test ,49,50. AThe behavior of subjects was recorded for 20 min with a camera  placed 150 cm above the arena. Behavioural performance from the video recordings was analyzed with EthoVision XT software  which allowed us to track the fishes’ swimming behavior in any virtual zones of the arena. Different individual movement variables of interest were chosen to analyze the fishes’ explorative behavior ,52,53:  which were then identified as A and B. The fish were taken out of the −80 °C freezer where they were stored and kept overnight at a temperature of 1 °C to allow for slow thawing. After measuring the total and muscle lengths (standard length), each fish was weighed and the color of the skin on the fish’s left side was determined. Each fish was then dissected and the weight and color of both fillets (right and left) were recorded. The color of skin and fillets was measured in three positions , with a CHROMA METER CR-400/410  recording the color parameters L* (lightness), a* (redness index), and b* (yellowness index). After that, the following parameters were also calculated:Finally, the fillets were skinned and homogenized for chemical analyses.For the chemical analyses, because of the fish size, the minced fillets of each specimen A were combined with the minced fillets of the corresponding specimen B in order to obtain a single sample. The total lipids of fillets were determined. Lipids were extracted according to Folch et al.’s  method, To determine the oxidative state of the fillets, primary and secondary oxidative products were quantified as conjugated dienes (CD) and thiobarbituric acid reactive substances (TBARS), respectively. First, CDs were quantified by a spectrophotometric method by dissolving 0.1 μL of lipid extract in 3 mL of pure hexane. Then, using a Lambda spectrophotometer EZ 150 UV/VIS , the absorbance of the sample was detected at 232 nm. For the calculation of the conjugated dienes, a molar extinction coefficient of 25,200 mL/mmol was used and the results were expressed as the mmol Hp/100 g of sample. Subsequently, 2 g of homogenized fillets were utilized to determine lipid oxidation products (TBARS). TBARS were determined according to the spectrophotometric method previously proposed by Vyncke , which qFor each tank, liver and distal intestine samples obtained from 10 fish  were fixed in Bouin’s solution for 24 h at 4 °C and subsequently washed in 70% ethanol. Samples were dehydrated using 80, 95, and 100% ethanol solution, washed with xylene, and finally embedded in paraffin  . Paraffi2 surface area. The score assignment criteria for the above-mentioned parameters are reported in The evaluation of distal Intestine morphology and the assignment of histopathological indexes were performed on three transversal sections per fish , taken at 200 μm intervals and stained with: (i) Mayer’s hematoxylin and eosin Y  for mucosal fold height, submucosa width, mucosal fold fusion, basal inflammatory influx, and supranuclear vacuolization of enterocytes; and (ii) Alcian blue  for the relative abundance of Alcian blue positive (Ab+) goblet cells calculated on a 500 µmThree liver sections per fish  were taken at 200 μm intervals and stained with: (i) Mayer’s hematoxylin and eosin Y  to assess hepatocyte morphology and the degree of fat accumulation; and (ii) Periodic Acid Schiff  to highlight glycogen deposition in hepatocytes. Regarding the degree of fat accumulation, a quantitative analysis was performed on three sections per liver sample . Blood vessels and bile ducts were excluded from quantitative analysis since they are considered non-assessable areas. Results were expressed as the percentage of area occupied by fat on the total hepatic parenchyma analyzed in the respective section.Image analysis was performed using the ZEN 2.3 software , while the percentage of fat fraction was determined with ImageJ.2 optical windows  and allowed to air-dry for 30 min. Using a 15× condenser/objective, specific areas were detected on each section, on which infrared maps of 164 × 164 µm size  were acquired in transmission mode in the mid-infrared range . The background spectrum was acquired on a clean portion of the optical window before each sample acquisition. The Atmospheric Compensation (for the correction of CO2 and water vapor atmospheric contributions) and the Vector Normalization (applied on the full frequency range to avoid thickness variations) routines  were used to pre-process all the raw infrared maps.For each tank, liver samples obtained from 5 fish  were collected and stored at −80 °C for Fourier Transform Infrared Imaging (FTIRI) analysis. A Bruker Invenio interferometer coupled with a Hyperion 3000 Vis-IR microscope and equipped with a Focal Plane Array (FPA) detector operating at liquid nitrogen temperature  was used for FTIRI measurements. Three sections of 10 µm, taken at 200 µm intervals using a cryotome , were deposited on CaF−1 , 1755–1720 cm−1 , and 1082–960 cm−1 .To evaluate the biochemical composition of hepatic tissue, false-color images were obtained from the integration process of infrared maps in the 2995–2820 cm−1 and 1800–900 cm−1 spectral ranges.For each IR map, the integrated areas of the spectral ranges mentioned above were also calculated and used to obtain the following band area ratios: LIP/TBM, FA/LIP, and GLY/TBM; TBM represents the sum of the integrated areas calculated in the 3050–2800 cmTotal RNA extraction from distal intestine samples obtained from 5 fish per tank  was performed using TRI Reagent . The final RNA concentration was obtained with a Nanophotometer P-Class , while the RNA integrity was checked by 28S and 18S ribosomal RNA band staining (GelRed™) on 1% agarose gel. The cDNA synthesis was performed with the iScript™ cDNA synthesis kit  using 1 μg of total RNA.http://www.ncbi.nlm.nih.gov, accessed on 9 February 2023) and are summarized in Real-time qPCR were performed in an iQ5 iCycler thermal cycler  using a 96-well plate according to Chemello et al. . For eacp < 0.05, and results were expressed as mean ± standard deviation (SD). Data on explorative behavior  did not meet the assumption of normality and/or homoscedasticity. The non-parametric Kruskal–Wallis test was used for analyzing each set of behavioral data and the results were expressed as first quantile (Q1)—median—and third quantile (Q3). The statistical software package Prism-8  was used for the data analysis.The aquaponics systems were used as the experimental unit for data related to survival rate and zootechnical performance, while fish were considered the experimental unit for all the remaining analyses. All data were checked for normality (Shapiro–Wilk test) and homoscedasticity (Levene’s test). Data on ∆breathing frequency, fillet quality, histological analysis, spectroscopic analyses, and relative quantification of gene expression were analyzed using a one-way analysis of variance (ANOVA) followed by a Tukey’s multiple comparison test. Significance was set at In all the experimental groups, ammonia and nitrite concentrations were lower than 0.05 mg/L at each measurement. No significant differences were observed in terms of nitrate  and phosphate  concentrations among the experimental groups.At the end of the feeding trial, the survival rate (SR) of the fish was 100% in all the experimental groups and all the feed provided was completely consumed by fish in all experimental groups. Zootechnical performance is reported in The analysis of each behavioral measurement during the open-field test, i.e., distance covered, immobility, time exploring the center of the arena, mean turn angle, and ∆breathing frequency, did not show significant differences among the experimental groups .p < 0.05). In addition, the weight (and percentage) of the intestine and spleen together was also significantly different between the HPM0 and HPM20 groups (p < 0.05). The skin lightness (L*) of the HPM0 group was higher than that of the other experimental groups. On the contrary, the highest fillet lightness value was evidenced by the HPM20 group. Finally, the HPM3 group showed the lowest (p < 0.05) lightness values of both skin and fillet in comparison with the other experimental groups.The physical characteristics of European seabass juveniles were not significantly affected by dietary treatments, except for the fillet yield, the intestine/spleen weight, and the lightness index (for both skin and fillet) . First, p < 0.05) lower SFA content compared to the other experimental group, while the HPM3 group was characterized by a significantly (p < 0.05) higher n6 PUFA content compared to the HPM0 and HPM20 groups. As regards the fatty acid profile of the fillets, the HPM20 group showed a significantly (p < 0.05) lower content of palmitic (C16) and stearic (C18:0) acid. Finally, the linoleic acid (C18:2n-6) content was significantly (p < 0.05) higher in the HPM3 group compared to both the HPM0 and HPM20 groups.The results of lipid oxidation  revealedDistal intestine samples from all the experimental groups did not show major signs of inflammation or alterations of histological architecture a–c. The p < 0.05) higher value compared to both the HPM0 and HPM3 groups . Finally, the HPM20 group showed a slight decrease in glycogen deposition compared to the other experimental groups , and a significantly lower amount of glycogen  compared to the other experimental groups, which did not show significant differences between them.Considering the expression of genes involved in the immune response, no significant differences were evident among the experimental groups .The physiological responses to different HPM dietary inclusion levels have been widely explored in several key fish species for the Mediterranean aquaculture, including rainbow trout ,61 and gA desirable goal in modern aquaculture is the use of full-fat HPM, due to: (i) the high cost associated with the HPM defatting procedures; and (ii) the possibility to improve the quality of insects’ biomass by modulating the composition of their growth substrate with potential sources of long-chain PUFA using macroalgae or dried microbial biomass ,34,66. AThe European seabass represents one of the most important commercial species in the Mediterranean area and, as an euryhaline fish, has the potentiality to be reared in very-low-salinity aquaponic systems coupled with several commune plant species . This apThe dietary full-fat HPM inclusions tested in the present study did not have negative effects on fish survival and zootechnical performance, indicating that the experimental diets were well formulated and accepted by the fish. In addition, no behavioral alterations were observed among the experimental groups. These positive outcomes were expected, since no cognitive decline has been recorded in fish fed HPM-based diets  , and mosIn this context, the histological analyses performed in the present study did not show major changes in histological architecture or signs of inflammation in distal intestine samples from all the experimental groups. These results are supported by the molecular analyses of immune-response-related gene expression, which did not reveal significant differences among the experimental groups. These findings are in accord with previous studies that showed how even higher HPM dietary inclusion levels did not alter fish gut health status ,73,74. ILiver is one of the major storage sites of lipids, and particularly fatty acids, in many marine fish species, including European seabass . The maiL*), a particular feature that could influence consumer acceptance of the fish [b* to be the most affected index by the addition of HPM in diets for rainbow trout [The fatty acid profile of full-fat HPM diets can also alter the fatty acid composition of the edible fish portion. Interestingly, a decrease in SFA content was observed from the HPM0 to HPM20 groups in the present study. Additionally, no significant changes in MUFA, n3, and n6 PUFA, as well as of important fatty acids, such as EPA and DHA, were evidenced in the fillets from fish fed an HPM20 diet compared to those of the HPM0 group. These results are consistent with a previous study on rainbow trout that used diets with similar inclusions of spirulina-enriched HPM . Furtherthe fish ,88, has ow trout , and witow trout , which dThe dietary inclusions of spirulina-enriched HPM tested in the present study ensured proper growth performance and did not cause behavioral alterations in European seabass juveniles. The bioactive molecules typical of HPM, such as chitin and lauric acid, as well as the ones derived from the spirulina dried biomass used to enrich the insects’ growth substrate (tocopherols and carotenoids) confirmed their key role in preserving the gut health status and the quality features of the fish edible fraction.As a general conclusion, the results obtained in the present study highlighted the suitability of enriched full-fat HPM as a proper sustainable aquafeed ingredient for European seabass rearing and the use of aquaponic culturing systems to meet the requested criteria for a more sustainable aquaculture.


The present study aimed to determine the association between sleep quality, light exposure at night, and psychological wellbeing in the 2n = 169). Sleep quality, light exposure at night, and psychological wellbeing were self-reported using the Pittsburgh Sleep Quality Index (PSQI), Harvard Light Exposure Assessment (H-LEA), and Depression, Anxiety, and Stress Scale (DASS-21) in the 2nd trimester and followed-up at the 3rd trimester.This prospective study was conducted in 9 randomly selected government maternity clinics in Kuala Lumpur, Malaysia. Healthy women aged 20–48 years old with single pregnancy were recruited using convenience sampling , anxiety (42 and 44.3%), and depression (9.6 and 16.6%) were observed among the participants. Adjusted multiple linear regression revealed that poor sleep quality and higher light exposure at night were attributed to greater stress and depression symptoms in the 3rd trimester. Higher lux level exposed from 10 pm to < 1 am was associated with increased stress  and depression . Only poor sleep quality was observed to adversely affect anxiety  and depression levels  in the 2nd trimester.During the 2Present study provided preliminary findings on the association between sleep quality, light at night, and psychological wellbeing of pregnant women. As a recommendation, future research could investigate whether public health interventions aimed at decreasing artificial light at night can benefit sleep quality and the psychological health of pregnant women. Psychological wellbeing is the concept of optimal mental functioning that improves the overall health of individuals. Compromised psychological wellbeing continues to be a prevalent issue faced by pregnant women in countries worldwide, with issues such as heightened stress, anxiety, and depression symptoms. Globally, the prevalence of stress, anxiety, and depression among pregnant women ranged between 23–56.8%, 37.5–63%, and 19.9–45.2`% respectively [Throughout the years, research has documented numerous factors that play critical roles in affecting psychological wellbeing during pregnancy. Particularly, sleep, a biological process for physiological wellness, was extensively studied. Past studies elucidated that poor sleep quality, short sleep duration, and sleep disturbances were linked with negative psychological outcomes among pregnant women , 7. MoviUndoubtedly, artificial light usage has increased tremendously with modernization and technological advancement, especially at night. As the main exogenous zeitgeber to the body’s internal clock, light influences the sleep–wake cycle and circadian rhythm. When light reaches the eye and the retina, it is detected by the retinal non-image forming photoreceptors and in turn, the light perceived is transmitted to the master clock suprachiasmatic nuclei (SCN) neurons . SubsequAlthough artificial light at night has brought convenience to our lives, improper/excessive exposure to artificial light out of sync from the natural light–dark cycle may bring about negative consequences including circadian misalignment that suppressed melatonin and other physiological processes, increased risk of sleep disorders, and compromised mental health . For exaWhile the relationship between light exposure and psychological wellbeing is not fully understood yet, a plausible mechanism was proposed including the effect of light on circadian system, which in turn, affects important biological rhythms in the body that may affect mood-related behaviours during pregnancy . The cloInterestingly, a recent study has done a comparison of sleep quality and perceived stress, accounting for light exposure before bedtime among pregnant and non-pregnant women . Howevernd and 3rd trimesters [nd and 3rd trimesters of pregnancy. Study findings would allow for the identification of trends and critical timing during pregnancy for interventions to be implemented.Although past studies had investigated sleep quality and its association with psychological wellbeing among pregnant women throughout 2imesters , the rolrd trimester. Data collection was conducted from July 2019 to April 2021. Pregnant women were recruited from 9 government maternity clinics in Kuala Lumpur, Malaysia, selected using simple random sampling. The participants were recruited using a convenience sampling method during their antenatal check-ups at the selected maternal clinics. Those who were Malaysians aged between 20–48 years old and literate in English or Bahasa Malaysia were recruited for the study. A minimum sample size required was 69 participants, calculated based on the probability of poor sleep quality among pregnant women with (p = 0.18) and without antenatal depression (p = 0.39), with confidence level at 95% and 80% power [nd trimester (53% success in the study recruitment). Reasons for declining participation include lack of time and lack of support from family attributing to the commitment required for the research, particularly during the critical period of pregnancy. A total of 115 follow-up data was collected during the 3rd trimester, giving rise to a 31.9% loss mainly due to miscarriage, loss of contact due to unknown reasons, and movement control order related to Covid-19 pandemic where residents were encouraged to stay at home for non-essential activities. While online options were provided during the pandemic, some declined to participate due to the other negative implications related to the pandemic, as well as a lack of digital literacy.This was a prospective cohort study conducted in the 2nd trimester and followed up during the 3All procedures performed were in accordance with the ethical standards of the Declaration of Helsinki. All experimental protocols were approved by the Medical Research and Ethics Committee (KKM/NIHSEC/P19-125) and National Medical Research Registrar (NMRR-18–3412-45225), to which ethical standards set by the committee were met. Approval letter from the Health Department of Kuala Lumpur and Putrajaya was obtained to conduct research in government maternity clinics. Besides, a written informed consent form was collected from the participants prior to data collection.In the maternity clinic, sociodemographic details  were collected via self-administered questionnaires. Medical history such as previous birth complications and physical impairments were cross-checked with antenatal health records.The sleep quality of pregnant women was measured using the Pittsburgh Sleep Quality Index (PSQI), which evaluated the sleep quality for the past month . PSQI isnd and 3rd trimesters of pregnancy.Harvard Light Exposure Assessment (H-LEA) assessed light exposure at each hour for 3 consecutive days with 2 weekdays and 1 weekend day . The queDepression, Anxiety, and Stress Scale – 21 (DASS-21) was used to assess the levels of depression, anxiety, and stress of pregnant women over the past week . The quend and 3rd trimesters. The association between lux levels and sleep quality with psychological wellbeing during pregnancy was then evaluated using multiple linear regression adjusted for maternal age and household income. The observed association was expressed as beta coefficient (β) corresponding to 95% confidence intervals (CI). The effect size for paired samples t-test (Cohen’s d) and multiple linear regression analysis  were assessed accordingly.Statistical analysis was performed using SPSS version 23 software . The normality of data distribution was determined using the Shapiro Wilk test. Continuous variables were presented in mean ± standard deviation, while categorical variables were presented as frequency (percentage). We performed a paired samples t-test to compare sleep quality, psychosocial wellbeing, and light exposure (lux levels) among pregnant women in their 2The characteristics of participants were presented in Table nd to 3rd trimester, their sleep quality deteriorated (Table p = 0.046), sleep disturbance , and sleep duration score  with small yet non-trivial effects.As pregnant women progressed from the 2nd and 3rd trimesters. However, it should be noted that the lux level generally increased from the 2nd to the 3rd trimester at all time blocks, despite insignificant increases from 7 pm to < 4 am. Moreover, the lux level exposed at the time block 4 am to < 7 am was significantly higher in the 3rd trimester , with a small non-trivial effect size.In terms of light exposure at night, the highest lux exposed was at 7 pm to < 10 pm, followed by 10 pm to < 1 am, and the lowest exposure at sleep time 1 am to < 4 am  and depression . On the other hand, there was a lack of association between lux level and the psychological wellbeing of pregnant women in the 2nd trimester (Table p = 0.002) and depression levels  in the 2nd trimester with small yet non-trivial effects.Adjusted multiple linear regression revealed that sleep quality measured by global PSQI score and lux level were consistently attributed to greater stress and depression symptoms in the 3nd and 3rd trimesters. As reported earlier [In the current study, we assessed the psychological wellbeing, sleep quality, and light exposure level at night in 2 earlier , stress, earlier . Indeed, earlier .rd trimester of pregnancy. Undoubtedly, the relationship between sleep, light, and psychological wellbeing is complex, and it is not possible to establish the causal effect based on the study design. However, several hypotheses could be raised based on the findings. Firstly, light exposure at night could adversely affect the circadian rhythm [Current study findings showed the associations of light exposure and sleep with psychological wellbeing during pregnancy in the 3n rhythm , which rn rhythm . Environn rhythm .Several studies conducted thus far could help shed light on the relation between light exposure, circadian misalignment, and subsequently, poor psychological wellbeing. First, it has been shown that approximately 6.5 h exposure to 100 lx level at night can produce one-half of the response for a stimulus almost 100 times brighter (9000 lx), suggesting that the circadian rhythm is highly sensitive to light intensity and duration exposed, thereby generating significant phase-delaying effect and melatonin suppression . A preliIn terms of wavelengths of light, a recent randomized-controlled trial among pregnant women elucidated that the use of blue-blocking glass 3 h before bedtime had favorable effect on the circadian rhythm with advanced melatonin onset and increased melatonin levels at night . PromisiOn the other hand, light exposure at night could act as a stressor to pregnant women which disrupted sleep and mood, contributing to the feelings of stress. For instance, when healthy participants were exposed to light (100 lx), subjective sleep quality and rapid eye movement (REM) sleep decreased, while light sleep and sleep awakening increased with lower nocturnal melatonin level and poorer psychological wellbeing , 37. In Recently, the American Medical Association issued a policy statement based on mounting research evidence on the effect of artificial light on human physiology with direct connections to health, including mental wellbeing, mood, and depression . True tond to 3rd trimester in the current study, often reported in longitudinal studies [In terms of sleep quality, the global PSQI score of participants in the present study was comparable with a recent meta-analysis comprising 24 studies and a total of 11,022 pregnant women. The meta-analysis reported an average score of 6.09 , corresp studies . Such fiOur findings add to the body of literature on the association between negative sleep quality and poor mental health in pregnant women . The relnd and 3rd trimesters of pregnancy, while more than half did not exhibit anxiety symptoms, thereby limiting potential associations. Furthermore, we found no significant association between light exposure in the 2nd trimester with psychological wellbeing in 3rd trimester . It seemed that the influence of light on psychological wellbeing is acute in the present preliminary findings and perhaps, longer exposure, follow-up periods, and objective assessments would provide better insights. We also did not assess possible comorbidity with other conditions such as substance use disorders and the link with light exposure as this was beyond the scope of current study. In addition, the study was conducted in the capital of Malaysia, Kuala Lumpur. While we did not recruit pregnant women from all states in Malaysia, Kuala Lumpur remained one of the most populous and fastest-growing cities in the country, reflecting the multicultural nation with citizens originating across all states in Malaysia [The present study has several limitations to note. Owing to our study design, it is possible that those who exhibit symptoms of stress and depression tend to sleep late with increased exposure to light at night. Further, we did not account for light exposure throughout the day, precluding a comprehensive characterization of 24-h light exposure, which could affect sleep outcomes and psychological wellbeing . AdditioMalaysia . NeverthMalaysia , 49, andIn conclusion, the study provided preliminary findings that light exposure at night, sleep quality, and psychological wellbeing were closely linked. Study findings highlighted the need to improve sleep quality and to boost lighting strategies for the promotion of emotional care during the critical period of pregnancy. In turn, this could have long-term implications on maternal and child outcomes. As a recommendation, future research could investigate whether public health interventions aimed at decreasing artificial light at night can benefit sleep quality and the psychological health of pregnant women.


Data on demographic and anthropometric measures were extracted from medical records preoperatively and 6 and 12 months post-surgery. The variables assessed were: probing pocket depth (PPD), clinical attachment loss (CAL), bleeding on probing (BOP), and stimulated whole-mouth saliva. In this study, saliva samples were analyzed by electrophoresis and immunoblotting. The ELISA kit was used to measure the MMP8 levels to determine potential markers for obesity. Adiponectin and albumin levels were also evaluated. Weight loss was associated with significant changes in patients’ periodontal clinical data. Although 7 out of 10 periodontal patients showed an increase in salivary adiponectin levels after root planning treatment, when analyzed by Western blotting, the increase was not statistically significant . There was no correlation between albumin levels and salivary adiponectin pre-surgery, nor 6 months or 12 months after surgery. Weight loss was not improved by low-grade inflammation in bariatric patients, since albumin levels were similar between periods. Periodontitis is an inflammatory disease that is modulated by several factors, among which adiponectin plays an important role for the treatment of periodontal disease.This study analyzed the salivary proteomics, adiponectin and albumin, related to weight loss and periodontitis in patients undergoing bariatric surgery. This study included fourteen patients with morbid obesity (body mass index, BMI > 40 kg/m Obesity is a chronic disease that has increased substantially in recent years and predictions suggest that 58% of adults worldwide will be overweight or obese by 2030 . Morbid In patients with type 2 diabetes mellitus, metabolic syndrome and coronary artery disease, reduced plasma levels of adiponectin may play a key role in the development of insulin resistance. Although the mechanisms underlying the anti-inflammatory properties of adiponectin are not well understood, the anti-atherogenic and anti-inflammatory properties of adiponectin may be related in part to its ability to stimulate nitric oxide production by the vascular endothelium . AdiponeIn this sense, saliva plays a key role in protecting and maintaining the integrity of the oral mucosa through lubrication, buffering action, antibacterial and antiviral activity, and food digestion . ProteomPeriodontal disease is a chronic disease characterized by pathogenic levels of periodontal pathogens, as a result of inflammatory and immune-mediated tissue changes . The bacCollagenase-2 (MMP8) is a tissue-degrading host proteinase secreted by neutrophils and fibroblasts. It has a role in maintaining a healthy periodontium  and is eTherefore, the relationship between periodontal disease and obesity has been widely discussed in the literature ,18. ObesTo date, little is known about the association between weight loss and adiponectin and albumin levels in periodontal patients. The hypothesis of this study was that weight loss would increase salivary adiponectin and albumin levels, which may impact the oral health of periodontal patients. Salivary levels of MMP8 were used as markers of gingival health.2 [This is an observational, longitudinal study with morbidly obese patients who were under multidisciplinary follow-up to undergo bariatric surgery treated at the Brazilian Unified Health System (SUS). Obesity was assessed using the body mass index (BMI). Values were obtained by accessing weight in kilograms and height in meters and centimeters. Patients were classified as morbidly obese if they had a BMI from 40 kg/m2 .In a previous study, it was demonstrated that anthropometry and DXA were equally useful in obese women, whereas anthropometry had no predictive power and DXA was the only acceptable predictor in obese men . Our samPatients were recruited from April 2010 to March 2012, treated at the School of Medicine Hospital, São José do Rio Preto, State of São Paulo, Brazil. Individuals with less than six teeth, smokers, pregnant women, carriers of infectious diseases and medicated with anti-inflammatories or antibiotics at least 3 months before this study were excluded. Therefore, from the total initial sample, 14 patients underwent Roux-en-Y gastric bypass (RYGB) bariatric surgery and were reassessed 6 and 12 months after surgery.This study was conducted in accordance with the Declaration of Helsinki, and was approved by the Ethics Committee for Research with Human Beings of the School of Medicine Hospital–FAMERP (process nº 0315/08). Written consent was obtained from all subjects prior to the start of this study, after being informed of the study objectives.Complete medical histories were obtained from the patient’s records and confirmed by interview. Health information, C-reactive protein (CRP) and glucose levels were obtained from the patient’s medical record.One dentist who had been trained at the beginning of this study performed all the examinations (Kappa > 0.80) for intra-examiner concordance.The measurements were performed with the use of a standard periodontal clinical probe 15 with a cylindrical shape, 15 mm scale, 1.75° cone taper, and 0.5 mm probe tip diameter  for the clinical examination of all present teeth. Periodontal condition measures including bleeding on probing (BOP), presence of calculus (PC), probing depth (PD), and clinical attachment loss (CAL) were assessed at baseline, month 6 and month 12 after bariatric surgery .The gingival and calculus indices were used to measure the presence of bleeding or calculus, respectively. Bleeding on probing indicates the presence of gingivitis. PPD was measured as the distance between the gingival margin and the bottom of the gingival crevice. CAL was determined by measuring the distance between the cement–enamel junction and the bottom of the gingival crevice. Sites with PPD equal to or greater than 4 mm and with CAL equal to or greater than 3 mm constituted the presence of periodontitis . All patOnce samples were taken, all individuals underwent full-mouth scaling and root planning (SRP) and participants were given standard oral hygiene instructions. For daily dental hygiene the brushing technique used was Bass modified, with a soft brush, twice a day. With supragingival scaling and polishing, this significantly improved the gingival bleeding.g at 4 °C for 15 min to remove all debris [Saliva sample collection was performed at three different times, using a standardized saliva collection protocol. All participants were instructed not to ingest food or drink for at least 1 h before sample collection. Initially, they rinsed their mouths with 5 mL of potable deionized water, and then were asked to swallow saliva for 5 min. After this period, patients were instructed to chew a small sheet of parafilm and spit out all the saliva accumulated in the mouth into a plastic tube immersed in ice for 10 min (stimulated flow). When all saliva samples were obtained, they were thawed at room temperature, divided into 1.5 mL tubes, and centrifuged at 14,000× l debris . The supSaliva samples were prepared for electrophoresis by dilution in sample buffer of 4× concentration ) with the addition of 0.5 M dithiothreitol  to the sample buffer solution. Electrophoresis was performed according to the manufacturer’s instructions using 4–12% Bis-Tris gels (Life Technologies Ltda.) and MES running buffer. Separated proteins were electrotransferred to nitrocellulose membranes, stained with FITC and photographed under UV light (350 nm). The blots were probed with an affinity isolated antibody fraction of rabbit antiserum to a synthetic human adiponectin peptide corresponding to amino acid residues 225–244 (Sigma-Aldrich), an immunogen affinity purified fraction of antiserum for a synthetic peptide from the human MMP8 hinge region  or monoclonal anti-human serum albumin derived from HSA-11 hybridoma (Sigma). Binding was detected using an affinity purified goat-ant-rabbit IgG labeled with peroxidase  and then the Clarity TM Western ECL substrate detection system . Chemiluminescence was detected by the ChemiDoc TM MP Imaging System  Molecular masses were determined by comparison with standard proteins.MMP8 was included in this study to investigate the health of periodontium to relate the clinical evaluation. In this way, MMP8 levels in patients’ periodontal samples were measured by ELISA. A volume of 100 µL mouse anti-human MMP8 (R&D Systems) at 2.0 µL/mL diluted in PBS per well was incubated in a 96-well plate and incubated overnight at room temperature. Three washes were performed in phosphate-buffered saline with 0.05% tween (PBS-T). A volume of 300 µL of phosphate-buffered saline with 1% BSA was added to each well and allowed to incubate at room temperature for one hour, followed by three washes with PBS-T. A serial dilution of 100 µL of samples and standards was performed in duplicate and the plates incubated for 2 h at room temperature, followed by 3 washes with PBS-T. A volume of 100 µL of goat anti-human MMP8 (R&D Systems) at 50 ng/mL diluted in phosphate-buffered saline with 1% BSA was added to each well and allowed to incubate for 2 h at room temperature, followed by three washes with PBS-T. A volume of 100 µL of 1:200 streptavidin-HRP in phosphate-buffered saline with 1% BSA was added to each well and allowed to incubate for 20 min at room temperature, followed by three washes with PBS-T. A volume of 100 µL 1:1 H2O2 to Tetramethylbenzadine Substrate Solution added to each well and left to incubate for 20 min at room temperature followed by addition of 50 µL 2MH2SO4 Stopping Solution. Absorbance was read at 450 nm using a plate reader  .p < 0.05. The analyses were performed using the JAMOVI 2.2.5 software. The protein will be analyzed by Scaffold software.Descriptive statistics  were calculated after verifying the normal distribution (Shapiro–Wilk). The analysis included ANOVA for repeated measures to verify the difference between the three periods (preoperative and 6 and 12 months post-operatively) followed by Tukey’s test for post hoc comparison. Pearson correlations were performed to assess the correlation between adiponectin and albumin levels preoperatively, 6 and 12 months after surgery. The significance level was set at A decrease in the bariatric patient’s BMI from preoperative levels at 6 months, and a further decrease at 12 months, indicates a successful weight loss intervention. This weight loss was associated with significant changes in the patient’s periodontal clinical data, with both clinical attachment loss (CAL) and pocket depth (PD) showing an increase after 6 months, indicating a worsening periodontal condition. At 12 months, there is a reduction in CAL and PD suggesting a recovery of the periodontal condition .Clinical evaluation indicated a reduction in periodontal disease. Although gingival recession increased, pocket depth and bleeding on probing decreased after surgery .Nine out of ten bariatric patients showed a decrease in salivary adiponectin levels 6 months after surgery compared to the preoperative sample . This cop = 0.084); however, when sample #37 was removed from the analysis, as it appeared to be an outlier, there was a significant decrease at 6 months after sampling (p < 0.05). There was an increase in levels from 6 months to 12 months, which were not different from preoperative levels .To determine whether salivary adiponectin levels were affected by gingival health, the periodontal disease of non-obese patients was analyzed. Analysis of their saliva samples revealed that, although seven out of ten periodontal patients had an increase in salivary adiponectin levels, when analyzed by Western blotting , the incSalivary levels of MMP 8 were used to assess patients’ gingival health. Five out of ten periodontal patients showed a decrease in MMP8 salivary levels during the period when analyzed by Western blotting (results not shown), which were not statistically different .p = 0.044). MMP8 analysis of saliva from bariatric patients was only possible using prior Western blots due to limited sample volumes. These results showed no difference in the preoperative period compared to the post-operative period (6 or 12 months).However, after analysis of the same samples by ELISA, eight of the ten patients showed a decrease in salivary MMP 8 levels  and a sip > 0.05)  .2 = 0.0456).Salivary adiponectin in samples from bariatric patients were plotted against pre-surgery salivary albumin levels, either 6 months or 12 months after surgery, there was no significant correlation  between pre-surgery salivary adiponectin levels and post-6 month levels and a non-significant increase (p = 0.2031) between the 6 month and 12 month levels. There were no clinical grounds for excluding this outlier as an anomaly, but its presence can be explained by the pre-surgical adiponectin level of the corresponding patient being almost too low. Weight loss was expected to lead to an increase in salivary adiponectin levels, as BMI levels show reduced obesity, which is correlated with higher adiponectin levels [The different modulation of adiponectin in saliva and serum of obese patients could be explained assuming that different sources of the adipokines detected in saliva may originate from both local production or may be transported by other biological fluids . Salivarn levels  and plasn levels  and fromn levels . Howevern levels . The retAnalysis of the clinical data shows a significant increase in pocket depth and clinical attachment level 6 months after surgery, followed by a decrease from 6 month to 12 months after surgery. The change in periodontal disease between baseline and 12 months was from 3.21 to 2.90 mm pocket depth, which is not clinical relevant. This suggests a worsening of the periodontal condition of patients undergoing Roux-en-Y gastric bypass surgery, followed by a return to a healthier periodontal condition, a negative correlation with salivary adiponectin levels.A previous study highlighted that MMP9 was associated with obesity and periodontitis  and MMP-Although hormones are known to be involved in albumin metabolism, no single hormone has been identified as a prominent factor in the physiological regulation of albumin metabolism. Albumin function is an indicator of nutritional status and actions of antioxidants. There is considerable knowledge about albumin synthesis, but the factors that regulate albumin degradation are not clearly established. The role of albumin itself in the synthesis of specific liver proteins is not fully understood, although in vitro data suggest that albumin may be a key factor. The apparent increase in the hepatic synthesis of specific proteins may be able to sustain not only colloid osmotic pressure, but also albumin transport and function .Periodontal microbes that trigger an inflammatory response, such as T. denticola, increase the levels of salivary albumin. Additionally, these microbes use albumin, as well as immunoglobulins, as potential energy sources . A recenElevated urinary albumin excretion independently and continuously predicts an increased risk of cardiovascular disease, even at levels below the threshold for the diagnosis of microalbuminuria. Albuminuria likely reflects systemic endothelial dysfunction rather than directly contributing to increased risk of cardiovascular disease . HoweverSalivary protein and albumin were positively correlated with periodontal patients, which demonstrated that the increase in salivary protein concentration in individuals with gingivitis or periodontitis was caused by plasma protein leakage . In our There were some limitations in the present study. First, the sample size was small; however, a causal relationship between weight loss and adiponectin levels of periodontal patients has been established. Second, the population of this study was the convenience sample (non-probabilistic sampling), which may have introduced some selection bias. Finally, there were limitations in the evaluation of saliva and plasma of albumin levels to compare between these biomarkers. However, there were several important strengths of our study that are also worth mentioning. This study examined the association of salivary albumin and adiponectin with the interaction between bariatric surgery and periodontal disease. In addition, important covariates were measured and controlled in the study analysis.Adiponectin is produced by fat cells and is considered an anti-inflammatory molecule. There was no correlation between salivary adiponectin levels and salivary albumin levels, which suggests that salivary adiponectin does not enter saliva through the same pathway as albumin, but may reflect local production.These findings provide insight that weight loss was not improved by low-grade inflammation in bariatric patients, since albumin levels were similar between study periods. Periodontitis is an inflammatory disease that is modulated by several factors, among which adiponectin plays an important role for the treatment of periodontal disease in candidates of obesity surgery, while the mechanisms and clinical application require further exploration.


This assessment was accomplished by comparing the Technetium-99 m methylene diphosphonate (seq). For each patient, bone SPECT and facial CT scans were registered and reconstructed to create fused SPECT/CT images. The 99mTc-MDP uptake levels in the TMJs were statistically compared among the four groups.The study included 17 patients  with suspected osteoarthritis of the mandibular condyle, totaling 34 temporomandibular joints (TMJs). Based on clinical and radiographic examinations, the TMJs were categorized into four groups: normal (group N), internal derangement (group ID), osteoarthritis (group OA), and osteoarthritis sequelae , mainly driven by the disparity between group OA and both group N (p < 0.001) and group OAseq (p = 0.048).The Fused SPECT/CT image can be an effective tool for evaluating degenerative changes in the mandibular condylar head. The technique demonstrated the ability to differentiate between normal TMJs and those with internal derangement, osteoarthritis, or osteoarthritis sequelae. This approach holds promise as a valuable method in clinical assessments of TMJ degeneration.The online version contains supplementary material available at 10.1186/s40902-023-00399-1. Osteoarthritis with degenerative change of mandibular condylar head is a group of overlapping distinct disease which may have different etiologies but with similar biologic, morphologic, and clinical outcomes . Osteoar99mTc-MDP) have been applied to identify the status of the local metabolic alteration in bone and joint. This was considered a useful technique which can upgrade the sensitivity of bone scan and used for the assessment of the transplanted bone for degenerative disease, avascular necrosis after surgery [Diagnosis of the osteoarthritis on temporomandibular joint (TMJ) has mainly performed on clinical findings using the research diagnostic criteria for temporomandibular disorder (RDC/TMD) chart and plain radiography. Even though the plain radiography such as panoramic view and transcranial radiography are useful tool to diagnose osteoarthritis, it has limitation in detecting lesions of early stages, since lesions can be observed when there are 30 to 50% of changes in bone mineral mass , 5. The  surgery . However surgery . Therefo surgery . Through surgery –15.99mTc-MDP uptake ratio obtained in the fused image with the results of clinical and radiographic findings. The null hypothesis of this study was that the fused image of SPECT/CT would not be effective for the evaluation of degenerative change of mandibular condylar head.However, to the best of our knowledge, there were few studies on clinical usefulness of the fused image of bone SPECT and facial CT in the diagnosis of osteoarthritis of the TMJ. Therefore, the objective of this retrospective study was to evaluate the clinical effectiveness of fused image of SPECT/CT for the evaluation of degenerative change of mandibular condylar head by comparing the The fused SPECT/CT image of 34 mandibular condyles of the 17 patients  who visited the Department of Oral and Maxillofacial Surgery, Hanyang University Hospital, was retrospectively collected. All the 17 patients were suspected to have osteoarthritis of mandibular condyle on plain radiography and clinical examination; therefore, facial CT and bone SPECT were taken for further evaluation. The age distribution ranged from 14 to 66 years, with a mean age of 34.3 years. The protocol for this study was reviewed and approved by the Institutional Review Board (IRB) of the Hanyang University Hospital (HYUH 2012–11-009–012). The requirement for patient consent was waived by the IRB committee.Primary diagnosis was based on DC/TMD Axis I , the clinical findings , and bony change on plain radiography .And then facial CT and bone SPECT of the patients were acquired. Facial CT  was taken with exposure parameters at 120 kV, tube current 80 mA s/slice, and 1-mm slice thickness. Facial CT scan time was 10 s covering 17 cm. The estimated CT radiation dose was 6.5 mGy to each patient.99mTc-MDP (740 ~ 1110 MBq). The SPECT was taken on a dual head gamma camera  using a low-energy, ultrahigh resolution collimator in a 128 × 128 matrix in continuous mode for 64 views per detector over 180° for 20 s per view. The bone SPECT data were reconstructed using OncoFlash .The bone SPECT was acquired 4 h after the intravenous administration of Normal group (group N) included the TMJs defined as no symptom except occasional clicking sound and normal feature on radiographic exam.Internal derangement group (group ID) was defined as an abnormal relationship between the articular disc and the mandibular condyle, articular fossa, and the articular eminence. These group represented the symptom of joint noise during normal function, mouth opening limitations, temporary joint locking, and pain without any bony change on plain radiography. These TMJs belonged to the group II of DC/TMD Axis I.seq) were belong to the group III of DC/TMD Axis I. In these groups, bony change such as bone erosion, cortical thinning, sclerosis, osteophyte, and irregularity on condylar head was observed on radiographic exam. Group OA showed the symptom of arthralgia on TMJ, and group OAseq showed no clinical symptom except crepitus.Group osteoarthritis (group OA) and group osteoarthritis sequelae , and the right and left condyles were indicated as R and L, respectively .Normal gSingle rater, who was profession of the Department of Oral and Maxillofacial Surgery, assessed the TMJs and performed grouping, and another rater performed the grouping separately. The result of grouping coincided with each other.SPECT data and CT data for each patient were transferred and co-registered to yield fused SPECT/CT image in axial, coronal, and sagittal plan using 3D software  Fig. .Fig. 1Fa99mTc-MDP uptake level of the TMJ, a sphere shape region of interest (ROI) (3.0 × 3.0 × 3.0 pixel) was designated in the highest point for evaluation and the counts in both the condyles and clivus measured . Kruskal-Wallis test, and further multiple comparisons using Tukey test for the post hoc analysis, was conducted to evaluate differences between groups. Value of seq), respectively. Group OA showed the highest uptake ratio, and group ID was the second and followed by group OAseq and group N (99mTc-MDP uptake level among the 4 groups was observed (p = 0.000) and between group OA and group OAseq (p = 0.048)  Fig. . This reseq, and group N, which was consistent with the order of uptake ratio.Additionally, comprehensive condylar image with hot spot was observed through the fused SPECT/CT images Fig. . The scop = 0.000)  Fig. .Group ID was defined as an abnormal relationship between the articular disc and the mandibular condyle, articular fossa, and the articular eminence. This group represented the symptom of clicking sound during normal function, mouth opening limitation, temporary joint locking, and pain without any bony change. Six condyles belonged to this group. The mean uptake ratio in this group was 1.93, which was the second highest among the 4 groups (p = 0.000) and OAseq (p = 0.048) were observed in post hoc analysis   Fig. .Osteoarthritis, degenerative change of the mandibular condyle, is largely caused by the excessive loads on TMJ, for example, masticatory movement of hard and tough foods, injury, and bad habits. If the excessive loads last, it may cause dislocation of temporomandibular disc, which in turn may cause pain during mouth opening, joint sound, and mouth opening limitation. If this progresses further, bony change on condylar head can be developed –5, 7. At99mTc-MDP uptake ratio obtained in the fused SPECT/CT image of the 4 groups, classified according to the findings from the plain radiography and clinical symptoms. Several applications of fused SPECT/CT image were performed previously. One research suggested the usefulness of fused SPECT/CT image which improves the sensitivity of the unilateral condylar hyperactivity [99mTc-MDP SPECT has provided the quantitative analysis where the increased uptake ratio on condylar head reflects the activity of the mandibular growth [The objective of this retrospective study was to evaluate the clinical effectiveness of fused image of SPECT/CT for the evaluation of degenerative change of mandibular condylar head. This study performed a comparative analysis of activity . In the r growth . In oster growth .99mTc-MDP uptake ratio was demonstrated in patients with internal derangement, osteoarthritis, and osteoarthritis sequelae than normal group (p = 0.003) (p = 0.000) and between the group OA and group OAseq (p = 0.048) . Thi. Thi99mTThe result of this showed the possibility to distinguish whether it is in the stage of osteoarthritis (group OA) in the patients who show unclear bone resorption patterns on condylar head in a plain radiography or simply internal derangement (group ID). Appropriate clinically approach should be performed if detecting this before advancing to the severe degenerative change of condylar head. It is important to decide conservative approach or active joint movement therapy. If metabolic alteration on condylar head associated with acute inflammation was occurred, conservative approach should be preceded. Active joint movement therapy such as splint therapy or manipulation therapy would aggravate the degenerative change.Limitation of this clinical study was not using of larger sample size that could have yielded more generalized results. Nevertheless, if the sample size was expanded, it is anticipated that statistical significance between the groups could be achieved. This would not only allow for the determination of bony changes on the condylar head with or without inflammation but also aid in identifying whether the TMJ is in the initial stage of osteoarthritis or simply experiencing internal derangement. Diagnosing certain TMJ conditions solely based on a patient’s subjective expressions of symptoms can be challenging and sometimes unclear. In such situations, fused SPECT/CT images present a valuable diagnostic tool. The integration of SPECT and CT scans provides comprehensive insights, aiding in the assessment of the osteoarthritis status of the TMJ. By combining these imaging techniques, clinicians can obtain a more accurate and objective evaluation of the condition, leading to improved diagnostic accuracy and better-informed treatment decisions.Fused SPECT/CT image can be an effective tool for evaluating degenerative changes in the mandibular condylar head. The technique demonstrated the ability to differentiate between normal TMJs and those with internal derangement, osteoarthritis, or osteoarthritis sequelae. This approach holds promise as a valuable method in clinical assessments of TMJ degeneration.Additional file 1: Supplemental Table. Groups according to the clinical and radiographic findings, and values and comparison of 99mTc-MDP uptake ratio of the groups


Stenotrophomonas maltophilia) was the highest, whereas that of strain ZR5 (Delftia acidovorans) was the lowest. Overall, the obtained results indicated that ∼91.3% of the isolates were capable of producing at least one of the four hydrolytic enzymes. In addition, most of the tested strains produced ICC and TPC, which play a significant role in reducing stress in plants. The results of this study suggest that the tested endophytic bacterial strains can potentially be used to mitigate climate change-associated stresses in plants and to inhibit plant pathogens.Agricultural crops are exposed to various abiotic and biotic stresses that can constrain crop productivity. Focusing on a limited subset of key groups of organisms has the potential to facilitate the monitoring of the functions of human-managed ecosystems. Endophytic bacteria can enhance plant stress resistance and can help plants to cope with the negative impacts of stress factors through the induction of different mechanisms, influencing plant biochemistry and physiology. In this study, we characterise endophytic bacteria isolated from different plants based on their metabolic activity and ability to synthesise 1-aminocyclopropane-1-carboxylic acid deaminase (ACCD), the activity of hydrolytic exoenzymes, the total phenolic compounds (TPC) and iron-complexing compounds (ICC). Test GEN III MicroPlate indicated that the evaluated endophytes are highly metabolically active, and the best used substrates were amino acids, which may be important in selecting potential carrier components for bacteria in biopreparations. The ACCD activity of strain ES2 ( Agriculture plays a key role within the world economy. Agricultural production is a result of complex interactions amongst plants, soil, water, biodiversity, fertilisers, and agrochemicals. For the proper functioning of this complex network, all components need to be managed adequately . The FooTwo types of environmental stresses are classified, namely 1) abiotic and (2) biotic stress. Abiotic stress is caused by salinity, drought, flooding, cold and heat, heavy metals, and nutrient deficiency, amongst others, whereas biotic stress is a result of various organisms such as viruses, bacteria, fungi, nematodes, insects, herbivores, and weeds  abiotic . Savary Aphanomyces euteiches), onion white rot (Sclerotium cepivorum), wheat take-all (Gaeumannomyces graminis var. tritici), and wheat crown rot (Fusarium spp.) [Global climate change is a problem of high priority as it increases the frequency of extreme weather events and influences crop production worldwide. In addition to the direct effects of abiotic factors associated with changing climatic conditions, climate change can also increase the number of pests and weeds and the incidence of disease . Climateum spp.) . Moreoveum spp.) .One of the possible alternatives to reducing the different abiotic and biotic stresses is beneficial microorganisms that interact with plants . MicroorThe main mechanism used by bacteria to limit stress in plants is the reduction of ethylene levels through the hydrolysis of 1-aminocyclopropane-1-carboxylic acid (ACC), the precursor of ethylene in all higher plants, catalysed by the enzyme ACC deaminase (ACCD) . EthylenAnother mechanism used by bacteria for the direct protection of plants is the production of extracellular hydrolytic enzymes that attack the structural components of the cell walls of most fungi and play an important role in fungal mycelium degradation , 17. Man3+ forms insoluble hydroxides and oxyhydroxides). Plants and microorganisms have therefore developed active strategies for iron uptake. Bacteria obtain iron by secreting iron-compensating compounds. These bacteria play a particularly important role in an environment where the plant is exposed to iron deficiency. In addition, iron-complexing compounds mediate biological control mechanisms against phytopathogens, limiting their iron availability due to nutritional competition and, thus, inhibiting their proliferation and plant colonisation [Iron (Fe) is one of the essential trace elements involved in plant metabolism, and Fe deficiency can lead to abnormal respiration and photosynthesis. In soils, the total Fe content often exceeds the requirements of plants, but its bioavailability is insufficient under various environmental conditions  that are involved in increasing plant tolerance to stress.Zea mays L. (maize), Vicia faba L. (broad bean), Secale cereale L. (rye), Triticum aestivum L. (wheat), Arctium lappa L. (burdock), and Equisetum arvense L. (horsetail). A description of the isolation process and the results of their identification based on 16S rRNA gene sequencing are presented by Woźniak et al. [The bacterial strains used in this study were obtained from the bacterial collection of the Department of Agricultural Microbiology, Institute of Soil Science and Plant Cultivation, State Research Institute, Pulawy, Poland. Overall, 23 strains of endophytic bacteria were selected from the tissues of four different crops and two wild plants Table . Bacterik et al. . The biok et al. .Table 1LPure cultures of the endophytic bacteria were characterised by the Biolog GEN III system , following the manufacturer’s instructions. This method allows the establishment of a metabolic profile for specific microorganisms, i.e., a “phenotypic fingerprint”.4)2SO4 (0.2% w/v); (B) negative control—DF minimal medium (without nitrogen source); (C) DF + 3mM ACC  (as a sole nitrogen source). The plates were incubated at 28 °C for 72 h. The growth of bacteria on plates with the addiction of ACC was compared with that of the positive and negative controls. Growth on ACC (C) medium indicates the ability of the tested strains to use ACC as the sole nitrogen source and synthesise the enzyme ACC deaminase [Qualitative estimation was performed via the method described by Govindasamy et al. . To deteeaminase , 30.Broth, Sigma-Aldrich, USA). After overnight growth, the bacterial cultures were centrifuged at 10,000 × g for 10 min, washed with 0.1 M Tris–HCl (pH 7.5), and suspended in 5 mL of DF medium containing 3 mM ACC as the sole nitrogen source. The cultures were incubated for 48 h at 28 °C, and the biomass was collected by centrifugation at 10,000 × g for 10 min at 4 °C. The supernatant containing the cells was removed, and the cells were washed twice with 5 mL of DF salt minimal medium. The cells were then resuspended in a fresh culture tube in 7.5 mL of DF salt minimal medium supplemented with 3 mM ACC and incubated at 28 °C with shaking for a further 24 h (140 rpm/min.). The induced cells were then harvested by centrifugation at 6000 × g for 10 min and washed with 0.1 M Tris-HCl (pH 7.6). The contents of the tubes were centrifuged at 12,000 × g for 5 min, and the pellets were resuspended in 200 μL of 0.1 mol L−1 Tris HCl (pH 8.5). After washing, the bacterial cells were labilised by adding 10 μL of toluene and then vortexed at the highest speed for 30 s. Subsequently, 50 μL of toluenised cell suspension was placed in a fresh tube, and 5 μL of 0.5 M ACC was added, followed by shaking and incubating at 30 °C for 15 min. A 250-μL aliquot of the supernatant was transferred to a glass tube and spiked with 200 μL of 0.56 N HCl and 75 μL of 2,4-DNF  solution, and the contents were mixed and incubated at 30 °C for 30 min. Next, 500 μL of NaOH (2N) was added, and the absorbance of the obtained mixture was measured at 540 nm by using a Varian Cary 1E UV-visible spectrophotometer. The cell suspension without ACC was used as a negative control and that with (NH4)2SO4 as a positive control. The values of the concentration of α-ketobutyrate produced were determined by constructing a standard curve ranging from 0.1 to 1 mM. The ACC deaminase activity was calculated as the amount of α-KB produced in μmol mg−1 protein h−1. The total protein content was estimated using the Bradford method  according to the manufacturer’s protocol [The quantitative analysis of ACC deaminase activity was performed according to a modified method of Penrose and Glick , which iprotocol .The selected bacteria were screened for extracellular hydrolytic activity of protease, cellulase, lipase, and esterase in agar plate assays. For the assessment of protease activity, one loop full of endophytic bacterial strains was streaked on a skimmed milk agar plate containing 5% (w/v) skim milk . Proteolytic activity was observed by the development of a halo zone around the colony of endophytic bacterial isolates . For detThe concentration of total phenolic compounds (TPC) was determined in the supernatants of bacterial strains cultured on TSB medium for 48 h at 28 °C. The concentration of TPC was determined by reaction with Folin-Ciocalteu reagent . Spectro3+)-complexing compounds (ICC) was determined in the supernatants of bacterial strains cultured on TSB medium for 48 h at 28°C. For this, 300 μL of 6% FeCl3 × 6H2O in 1 N HCl was added to 1 mL of the obtained supernatants, and the samples were thoroughly mixed and incubated for 1 h at room temperature. Subsequently, spectrophotometric measurements (Varian Cary 1E UV-visible) were performed, determining the absorbance value at λ = 520 nm [The concentration of the total content of iron . The data from Biolog GEN III experiments were combined in a single matrix, represented as a positive integer, OmniLog TM units (OL units) and subjected to statistical analysis using the STATISTICA.PL (13.1) software  for presentation as a heat map graph. A multivariate statistical method using PCA was performed to summarise the variability of the tested strains and determine the association amongst the measured activities. Mean values from three replicates were used for PCA analysis. Prior to this analysis, the data were standardised.To investigate the overall metabolic profiles of endophytic bacteria, the substrates on the GEN III MicroPlate were categorised into six classes: sugars, sugar alcohols, hexose-PO, amino acids, hexose acids, and carboxylic acids. The results of the relative use of the chemical groups of subtrees are presented in Fig. All analysed strains showed the ability to grow on solid medium with ACC, albeit at different intensities Table . Based o−1 h−1 in DF minimal salt broth medium supplemented with ACC as nitrogen source. Only strain ER1 showed no ability to synthesise ACC deaminase. Moreover, strain ER1 showed the ability to grow on DF medium supplemented with ACC as a sole source of nitrogen, but it showed no ACC deaminase activity (Table −1 h−1) followed by ES1 (8.511 μmol α-KB mg protein−1 h−1), AR2 (6.446 μmol α-KB mg protein−1 h−1), and AR4 (5.505 μmol α-KB mg protein−1 h−1). These strains of endophytic bacteria belonged to the genera Stenotrophomonas (ES1 and AR4), Rhizobium (ES1), and Collimonas (AR2). By contrast, the lowest level of ACC deaminase production was observed in isolates ZR5 and ZR1 . Four of the tested isolates demonstrated the ability to produce ACC deaminase at different concentrations, both in medium with and without ACC. The addition of ACC as a nitrogen source significantly increased the efficiency of ACC deaminase biosynthesis.Amongst the 23 endophytic bacterial isolates, 22 strains (96%) showed variation in ACC deaminase activity in the range of 0.264–16.138 μmol α-KB mg proteinDelftia acidovorans, whereas the highest cellulase activity was detected in Delftia acidovorans isolate SR3 (EI = 5.583). Strain TS4 showed the highest lipase synthesis activity, and strain TS1 showed the highest esterase activity; both strains were classified as Delftia acidovorans.All bacterial isolates were screened for protease, cellulase, lipase, and esterase activities Table . PrelimiVariovorax paradoxus strain VR2 (576.516 μg mL−1). However, the contents of these compounds in the cultures of the other tested strains ranged from 6.015 to 316.779 μg mL−1  showed that almost all tested strains produced these compounds. Only strain AR2 showed no ability to produce ICC, whereas strains ZR3 and VS3 had the lowest concentrations of these compounds. The highest ICC level was found in the culture of the ES2 Fig. .Fig. 3CoDelftia and one strain from the genus Stenotrophomonas, isolated from crop plants. In contrast, strains collected from weeds were positively related to the ability to produce deaminase ACC and phenolic compounds. The PCA showed a separation of the evaluated strains based on their genus, resolving strains based on taxonomic affiliation and plant type (crop plants and weeds).The relationships amongst all endophytic bacteria and their properties were assessed via principal component analysis (PCA) Fig. . The firIn the context of the ecological and climate crises we are currently facing, along with the growing demand for food and the limitation of natural resources, agriculture must undergo a transformation towards maintaining balance and natural stability, biodiversity, and soil health . IncreasWith an increasing awareness of sustainable farming practices, it is important to seek region-specific microbial strains (native strains) that have a broad spectrum of activity and are compatible with other microorganisms in soil and plant tissues for the use as components of inocula to enhance plant resistance . The finSolanum lycopersicum and Arabidopsis thaliana [The Biolog GEN III MicroPlate test is based on phenotypic tests and allowed the assessment of the metabolic profile of selected strains of endophytic bacteria. Analysis of the metabolic activity provides important information about individual environmental isolates, which is crucial in understanding their functioning in future environmental niches, carriers, and biopreparations. The wider the spectrum of the compounds used, the greater the ability of the strains to survive under different conditions and compete with other potentially pathogenic microorganisms . In addithaliana . On the thaliana .nifH gene in our previous study [The ability of bacteria to synthesise ACC deaminase is one of the key growth-stimulating features affecting stress tolerance in plants under both normal and stress conditions . Accordius study .Stenotrophomonas, Rhizobium, Collimonas, and Delftia. The indicated isolates showed high ACCD activity with > 5 μmol of α-KB mg protein−1 h−1; according to Singh and Jha [1 h1 is sufficient to trigger systemic tolerance under stress conditions. The genus Stenotrophomonas has frequently been described as a genus with great potential in agriculture [Stenotrophomonas maltophilia SBP-9 augments resistance against biotic and abiotic stress in wheat plants through, amongst others, synthetising ACCD at the level 362 nmol of α-KB mg protein−1·h−1 [In the present study, the most active producers of ACC deaminase were strains of the genera  and Jha , the proiculture . For insin−1·h−1 .Rhizobium leguminosarum contributes to an increased efficiency of symbiosis with Pisum sativum L., both under single and complex stress conditions related to cadmium and water deficiency. Other authors demonstrated ACC deaminase activity in members of the genera Collimonas [Delftia [Belimov et al.  demonstrllimonas  and Delf[Delftia .Arctium lappa and Equisetum arvense, which are ubiquitous under various environmental conditions. In parallel with the crops, they grow and reproduce aggressively [Interestingly, the most effective producers of ACC deaminase were isolated from tissues of the weeds essively . Like alessively . In addiessively .Delftia and Stenotrophomonas showed all tested hydrolytic enzyme activities. Wang et al. [Delftia acidovorans strains isolated from different crop plants. According to the literature, strains of the Stenotrophomonas genus have a high hydrolytic potential to produce proteases, chitinases, glucanases, lipases, and other enzymes [S. maltophilia contribute to protecting plants from fungal pathogens such as Magnaporthe grisea [In this study, bacterial endophytes were qualitatively analysed for their ability to produce different hydrolytic enzymes. Overall, microbial synthesis of the hydrolytic enzymes appears to be important for the colonisation of plant roots and biocontrol efficacy , 17. In g et al.  emphasis enzymes . In parte grisea . These eAccording to a previous study, amongst endophytic bacteria, the highest hydrolytic activity is recorded against protease and cellulase . In cont3+ in the areas around the roots, preventing its assimilation by pathogens and, thus, facilitating disease inhibition [Variovorax paradoxus (VR2) strain was the strongest producer of compounds capable of complexing iron. In a similar study, Liu et al. [Variovorax boronicumulans strain promoted plant growth by producing siderophores, which was also correlated with the production of other secondary metabolites  for biofertiliser and biopesticide applications. In addition to plant growth stimulation and biological control, iron-complexing compounds, including siderophores, are also used in bioremediation [Agrobacterium radiobacter can reduce the As content in the soil by approximately 54% [The iron-complexing compounds are implicated directly by incentive nutrient uptake and indirectly by insulating Fehibition . In our u et al.  proved tediation  as they ediation , 64. Thiediation . SideropBacillus thuringiensis, phenols are crucial in controlling root-knot nematodes [Bacillus, Pseudomonas, and Methylophilus indirectly increased the total phenol content in tomato [C. pratensis (AR2), C. koreensis (ER1), A. xylosoxidans (AR3), and S. maltophilia (ES2), are a rich source of phenolic compounds, which translates into their potential use in the direct and indirect protection of plants against biotic stresses.Phenolic compounds comprise numerous groups of secondary metabolites with antibacterial and antioxidant properties. Built into the plant cell wall, they create structural barriers that prevent the development of the pathogen at the site of infection , 66. Altematodes . It was n tomato . Interesn tomato , based oDelftia bacteria isolated from cultivated plants can be suitable components of biopreparations with anti-pathogenic effects due to the positive correlation with the activity of hydrolytic enzymes. Moreover, bacteria isolated from weeds show a promising potential in mitigation abiotic stresses due to the positive correlation and high activity of ACC deaminases.The results obtained by the analysis of various properties of endophytic bacteria indicate that the complex features responsible for enhancing plant stress resistance are mainly determined by the type of plants and, partially, by the endophytic bacterium. Our previous studies based on PCA analysis clearly showed that the plant growth-promoting features of these endophytic bacteria were dependent on the bacterial genus, as identified by 16S rRNA gene sequencing . Jacoby Stenotrophomonas maltophilia) and ZS2 (Delftia acidovorans) as potential effective components of multi-task biopreparations . These strains showed high metabolic activity in the Biolog GEN III test, a high efficiency of N2 fixation and the ability to synthase indole-3-acetic acid (IAA) and siderophores [Comparison of the results presented in this manuscript with previously published findings  confirmsrophores . Our prerophores  showed trophores .Delftia spp.). Therefore, these bacteria can be the core of endophytic bacteria in a given area (core of native bacteria) and may have so-called “endophytic competences” related to the effectiveness of establishing interactions with plants. Our current and previous studies provide valuable information for the selection of the most promising endophytic bacteria for plant growth and protection. Strains ES2  and ZS2 (Delftia acidovorans), characterised by direct mechanisms of plant growth promotion and some of the highest deaminase and protease activities, showed the ability to produce ICC and TPC and use sugars, amino acids, and carboxylic acids. The strains of endophytic bacteria characterised here show potential for further research on the practical use as components of a multi-task biopreparation , which is in line with the assumptions of sustainable agriculture and environmental protection. In our future studies, the selected strains will be further evaluated in greenhouse and field experiments, in natural soil and under ambient climate conditions.In conclusion, amongst the tested strains of endophytic bacteria, a diverse ability to produce compounds positively affecting the growth and development of plants was observed. The evaluated strains were characterised by a wide range of mechanisms increasing stress tolerance in plants: lowering ethylene levels by producing ACC deaminase, degrading the cell wall of pathogens through the synthesis of hydrolytic enzymes and competing with pathogens through the production of iron-complexing compounds. An important aspect of this study is the identification of a relatively high number of bacteria that have a wide range of plant hosts (e.g.,