Furthermore, in the HP+BAs group plasma, CDCA, CA, LCA, and GCDCA increased significantly; they could activate TGR5, and the activation of hepatopancreas TGR5 might regulate glucose metabolism to relieve hepatopancreas glycogen accumulation. This study proved that BAs supplemented to plant protein diet could relieve the common carp hepatopancreas glycogen accumulation by changing the BAs’ profile, thereby promoting its healthy growth, which has important guiding significance for the promotion of aquaculture development and makes an important contribution to expanding the strategic space of food security.Bile acids (BAs) have considerable importance in the metabolism of glycolipid and cholesterol. The purpose of the present study is to clarify the effects of bile acids supplementary in a high plant protein diet for the common carp BA profiles and hepatopancreas and intestine health. An 11-week feeding trial was conducted with high plant protein diet  (HP) and HP added 600 mg/kg BAs (HP+BAs) for common carp, and then, the UHPLC-MS/MS technology was used to analyze the BAs in the bile and plasma of two groups. HP could induce vacuolation of hepatocytes and accumulation of glycogen in the common carp, while these phenotypes were significantly improved in the HP+BAs group. In addition, the BA profile of the HP group and HP+BAs group are described in detail, for the common carp bile with treatment by exogenous BAs, TCA, CA, T Most oftabolism ,14. ThesIn order to meet the needs of environmental protection and reduce the cost of feed, plant protein is increasingly widely used in aquatic feed . HoweverCyprinus carpio) [Common carp ( carpio)  is a kinDuring the feeding period, the experimental fishes were maintained in compliance with the Laboratory Animal Welfare Guidelines of China .β-muricholic acid-d4 sodium salt (TβMCA-d4), and tauroursodeoxycholic acid-d5 (TUDCA-d5) were obtained from Toronto Research Chemicals . β-muricholic acid-d5 (βMCA-d5) was bought from IsoSciences . Seven deuterium-labeled BAs containing deoxycholic acid-d4 (DCA-d4), glycolithocholic acid-d4 (GLCA-d4), glycoursodeoxycholic acid-d4 (GUDCA-d4), taurodeoxycholic acid-d4 sodium salt (TDCA-d4), glycochenodeoxycholic acid-d4 (GCDCA-d4), glycodeoxycholic acid-d4 (GDCA-d4), and ursodeoxycholic acid-d4 (UDCA-d4) were the products of Cambridge Isotope Laboratories Inc . LC-MS grade methanol, acetonitrile and formic acid were the products of Fisher Scientific. Other materials were obtained from Shanghai Anpel Laboratory Technologies . Bile acids supplementary products were supplied by Shandong Longchang Animal Health Care Co., Ltd., Dezhou, China (8.0% hyocholic acid (HCA), 70.9% hyodeoxycholic acid (HDCA), and 20.2% CDCA).Reference standards of unconjugated and conjugated BAs list in  includin18 column   C18-Aq GracePure TM (500 mg/3 mL), was the product of Grace Davison Discovery Sciences TM . The refrigerated centrifuge, Type 5430R, was bought from Eppendorf Inc, Germany. The Tissue Gnostics Fluorescence Imaging System was purchased from TissueGnostics .The ultra-high performance liquid chromatography-tandem mass spectrometry (UHPLC-MS/MS) utilized in the project was an Agilent 1290 Infinity II UHPLC coupled to an Agilent 6470A TripleQQQ (TQQQ) and AB SCIEX TripleTOF6600 (QTOF). The UPLC BEH CAs described in Yao et al.’s study , high plPlasma ALT , AST (aspartate aminotransferase), glucose, and total cholesterol (TC) were measured by Reagent kits  following the given protocols.The hepatopancreas tissue fixation, dehydration, embedding, hematoxylin and eosin (H&E) and periodic acid Schiff (PAS) staining procedures were conducted as described by Yu et al. . Then, t−). Both temperatures of ESI− source drying gas and sheath gas were 300 °C. The flow rate of ESI- source drying gas and sheath gas were 5 and 11 L/min, respectively. The pressure of the nebulizer was 45 psi, and capillary voltage was 4000 V. The dynamic multiple reaction monitoring (dMRM) was used to acquire data in optimized MRM transition (precursor -> product), fragment, and collision energy (CE) as 18 column . The column temperature was 40 °C, and the flow rate was 0.45 mL/min. The mobile phase consisted of water in 0.1% formic acid (A) and acetonitrile in 0.1% formic acid (B). The chromatographic separation was conducted by a gradient elution program as follows: 0.5 min, 15% B; 1 min, 25% B; 3min, 25% B; 5 min, 34% B; 8 min, 40% B; 9 min, 52% B; 10.2 min, 58% B; 10.21 min, 100% B; 11.2 min, 100% B; 11.21 min, 15% B; 12.5 min, 15% B. The gradient elution was applied and MS detection proceeded in negative mode. Standards for all BAs were used to identify the different BA metabolites detected by UHPLC-MS/MS. The Agilent Mass Hunter software (version B.08.00) was used to control instruments and acquire data. The raw data were processed by Agilent Mass Hunter Workstation Software (version B.08.00) by using the default parameters and assisting manual inspection to ensure the qualitative and quantitative accuracies of each compound. The peak areas of target compounds were integrated and output for quantitative calculation.Plasma and bile samples were prepared following the previous report . The eluβMCA and TωMCA were qualified by UHPLC (Agilent 1290)-Q-TOF (AB SCIEX| 6600)-MS/MS with an ESI source. The main parameters of ESI-MS/MS were as follows: declustering potential (DP): −100 v, collision energy (CE): −60 v, ion source gas1 (GS1): 50 arb, ion source gas2 (GS2): 60 arb, curtain gas (CUR):30 arb, temperature: 600 °C.Tm/z 50 to 1000 was acquired. PeakView 2.1 Software of AB SCIEX was used to analyze the ion fragment information of TβMCA and TωMCA standards and samples.Chromatographic separation was operated as 2.4. A mass range of t-test of variance by the software SPSS Statistics 20 was used to analyze all data. Homogeneity test of variance (F-test) was also performed for the data between the two groups; log transformation analysis was executed on the data when variance was irregular. Data are presented as mean ± SEM. Statistically significant results are indicated by asterisks . Graphics were drawn using GraphPad Prism 8.0 (GraphPad Software Inc. USA). Hepatosomatic index (HSI) was calculated by the formula of weight of the hepatopancreas (g)/body weight (g)∗100%. All BAs’ unit conversion was calculated from ng/mL to mM, and we summed individual BA concentration as total BA concentration (TBA). The average concentration of individual BAs and the total BAs’ concentration were normalized to calculate the ratio of individual BAs to the total BAs.Independent-samples p < 0.01) (p < 0.01), and HSI significantly decreased. The mean of HSI in HP and HP+BAs groups were 7% and 3%, respectively.Compared with the HP group, the growth performance of the HP+BAs group improved significantly ( < 0.01) , the finp < 0.001), while the HP+BAs group has a more active nucleus . Supplement BAs to a high plant protein diet not affected the content of TC in plasma. Plasma glucose and liver glycogen in the HP+BAs group were significantly lower (p < 0.05) than those in the HP group  and the abundance ratio of MS/MS. The RT and abundance ratio of MS/MS of Tnsistent .βMCA, GLCA, and HDCA increased significantly in the HP+BAs group, while TCA and CDCA decreased ) bile [−7 umol/mg. However, the fish meal in the experimental diets of this study only accounted for 10%, which made the bile acids level in the basal diet lower than the detection limit of LC-MS/MS. In the study of Staessen et al. (2021) [−5 umol/mg GBA were detected in the basal diet. In the present study, the dietary bile acid profile was composed with HCA (8.0%), HDCA (70.9%), and CDCA (20.2%), and the two experimental diets were designed with the same level of fishmeal. Hence, we can conclude that the increased GBA in the HP+Bas group should be majorly endogenous for the common carp. This is a report, for the first time, that bile acids can be conjugated with glycine in common carp, although it has been found that bile acids could hardly be combined with glycine in some fish species, such as Sea Lamprey [TCA was the main BA in the common carp bile, which was consistent with the results of previously reported studies about the BA profile of fish (angelfish (i)) bile . In thisi)) bile , mice [3i)) bile , and raii)) bile , we alsoi)) bile ,38, whic. (2021) , 27% fis Lamprey  and lake Lamprey . In addi Lamprey ,41.βMCA, GLCA, GCDCA, and HDCA in the common carp bile; we suppose the following could account for this with the help of intestinal microorganisms. HCA transformed into βMCA by 6b-epimerization and further 7b-epimerization [βMCA was reabsorbed back to hepatopancreas and combined with taurine into TβMCA, while HDCA was directly reabsorbed to the hepatopancreas. CDCA is dehydroxylated into LCA , which is partly excreted from the body, and part is reabsorbed to the liver and combined with glycine to form GLCA. The other part of CDCA is reabsorbed to the hepatopancreas to combine with glycine.Supplementary BAs  increased the contents of Trization , then βMβMCA and TωMCA in Common Carp was a breakthough since they were thought to be a rodent specific bile acid [The discovery of Tile acid ,44,45,46ile acid ,48, but ile acid ,49. RodeSupplement BAs reduced liver glycogen accumulation and alleviated liver damage in common carp with a high plant protein diet. High plant protein could induce fish intestinal and liver damage that has been confirmed in the previous research through our laboratory ,30,50,51Firstly, the soysaponins of soybean meal may be the main cause of common carp intestinal damage . BAs canβMCA inhibits intestinal FXR. TGR5 could be activated by some BAs, in which LCA is the most potent agonist for TGR5, DCA and the conjugations, CDCA and the conjugations, and CA and the conjugations activate TGR5 effectively simultaneously [Secondly, common carp hepatopancreatic inflammation and glucose metabolism may be regulated by three purposes that were LCA, CDCA, and CA to activate liver TGR, increased liver glycine concentration and Taneously . TGR5 planeously . TGR5 reaneously ,61,62. Aaneously . That suaneously , they wianeously . The expaneously .βMCA varied quite distinctly in bile between the HP group and HP+BAs group. TβMCA is a farnesoid X receptor (FXR) nuclear receptor antagonist [βMCA suppressed the enterohepatic FXR-FGF15 signaling and could affect glucose metabolism, reduce blood glucose, and treat diabetes [βMCA in bile after the addition of BAs also plays a certain role in keeping the hepatopancreas from avoiding histological damage and the reduction of plasma glucose.In addition, in the results of IDE et al. (1994), it can be found that the content of G-BAs in bile increases with the increase of liver glycine concentration . Kupffertagonist . FXR is tagonist . In the tagonist . TβMCA sdiabetes . TherefoβMCA and the proportion of G-BAs in bile increased significantly, which might play a leading role in it that reduced the accumulation of hepatopancreas glycogen and maintained hepatopancreas health. This study proceeded with an integrated bile acid profile determination by UHPLC-MS/MS to identify the effect of exogenous BAs supplementary on the endogenous BA profile and hepatopancreas health of common carp and discussed how the BAs supplementary is transformed in the body, providing a theoretical basis for the application of BAs products in fish and a data basis for revealing the mystery of fish BAs. In addition, this study has important significance for the development of aquaculture and has a potential contribution to expanding the strategic space of food security.In summary, HP could induce glycogen accumulation in common carp hepatopancreas while supplemented BAs to HP could mitigate this symptom. BAs supplements in a high plant protein diet could change the BA profile of common carp, among them, plasma LCA, CDCA, and CA increased significantly, T


There is a lack of high-quality evidence underpinning many contemporary clinical practice guidelines embedded in the healthcare systems, leading to treatment uncertainty and practice variation in most medical disciplines. Comparative effectiveness trials (CETs) represent a diverse range of research that focuses on optimising health outcomes by comparing currently approved interventions to generate high-quality evidence to inform decision makers. Yet, despite their ability to produce real-world evidence that addresses the key priorities of patients and health systems, many implementation challenges exist within the healthcare environment.This manuscript aims to highlight common barriers to conducting CETs and describes potential solutions to normalise their conduct as part of a learning healthcare system. Many recommendations contained in contemporary clinical practice guidelines (CPG) are not supported by high quality evidence . In AustComparative effectiveness trials (CETs) address this conundrum by randomising participants to usual care alternatives in order to generate unbiased, high-quality evidence of the relative effectiveness of existing treatments . Their pAs the treatment options investigated are widely used in practice, the additional risk to patients is low. Furthermore, public opinion polls repeatedly confirm that patients want their health systems to be ‘research active’ so that they are better informed about the benefits and risks of different treatments –6. It seMany factors have stymied attempts to integrate CETs into clinical care. Indeed, the characteristics that support their pragmatism can also hamper their conduct. For example, to preserve external validity, CETs necessarily avoid certain design features required for high internal validity,  . HoweverHealth system leaders too are reluctant to allocate resources to support trial activity, perhaps because the research agenda has historically been driven by academics and the trials conducted within their institutions have not always been in line with health system priorities.One of the most significant barriers to the adoption of CETs for contemporary evidence generation is the absence of proportionate approaches to trial governance.Currently, the policy framework and governance arrangements applied to CETs are largely identical to those for trials involving novel interventions. For example, some CETs compare interventions in which verbal consent to treatment would have been obtained outside the trial setting. Yet, when formally comparing two standard treatments, lengthy, legalistic consent forms are mandated, despite growing evidence that patients prefer simpler ways to be informed about such research that do not overinflate the perception of risk , 11. TheEfforts to develop, design and conduct CETs will continue to require creative approaches to overcome common challenges. Table The failure of health systems to ground everyday treatments in sound science impacts the quality of care provided to patients and contributes to health system waste. Conducting CETs that provide robust and generalisable evidence to guide practice is a public good. Importantly, patients recognise this and consistently confirm that they want their health systems to be research-active.Although some progress has been made, few healthcare organisations have successfully transitioned into LHSs that conduct trials as an integral part of the delivery of high-quality care. Despite progress in areas such as trial design, lingering challenges remain, not least, the need to develop risk-proportionate regulatory and governance frameworks that facilitate the conduct of large, pragmatic CET by them being less expensive and cumbersome to conduct.Implementing solutions that address the factors that currently prevent the embedding of CETs as a core activity of the health system will require disruptive change, but without such change, the widespread use of low-value or suboptimal treatments will remain unchecked.


Aenasius bambawalei is a predominant accompanying parasitoid of the important invasive pest Phenacoccus solenopsis. To understand the effect of temperature changes on the development of the reproductive systems of this parasitoid, we compared differences in its development, morphology, and fecundity under different temperatures. The results showed that high temperature could significantly shorten the pupal developmental duration, increase the length of the hind tibia, and accelerate gonad development. Our results indicated that moderately high temperature was conducive to the reproduction and development of the parasitoid and may be related to the more females offspring. This is the first report of the impact of high temperature on the pupal development, morphology, and fecundity of A. bambawalei.During biological invasions, insect pest outbreaks often occur because they escape the control of natural enemies from their place of origin. However, some natural enemies can migrate with pests to effectively inhibit their damage. Whether temperature changes can shorten or enhance the reproductive developmental period of accompanying natural enemies is an important determinant of whether they can effectively migrate with pests. Phenacoccus solenopsis (Homoptera: Pseudococcidae) is an aggressive invasive insect pest worldwide. Aenasius bambawalei (=Aenasius arizonensis Girault) (Hymenoptera: Encyrtidae) is a predominant accompanying parasitoid of this mealybug. Our previous studies showed that temperature change induced an increase in the female offspring ratio of A. bambawalei. However, whether this increase is the result of a shortened or enhanced development period of the reproductive systems of A. bambawalei remains unknown. Here, we compared the pupal development, hind tibia of female adults, and fecundity of A. bambawalei under different temperatures to clarify the development and morphological changes induced by high temperature and to better understand its potential as an accompanying natural enemy. Our results showed that, at a high temperature (36 °C), the pupal developmental duration of A. bambawalei was only 0.80 times that of the control, and the length of the hind tibia was 1.16 times that of the control. Moreover, high temperature accelerated the developmental rate of gonads and increased the numbers of eggs and sperm. These results indicated that experimental warming shortened the pupal developmental duration, altered the hind tibia length of female adults, and facilitated the fecundity of A. bambawalei. These findings will help to understand the adaptation mechanisms of accompanying natural enemies. Furthermore, these findings will help to make use of this behavior to effectively control invasive pests.The effects of high temperature on the developmental, morphological, and fecundity characteristics of insects, including biological invaders and their accompanying natural enemies, are clear.  High temperature may directly or indirectly influence the spread of alien species . Due to Phenacoccus solenopsis Tinsley (Hemiptera: Pseudococcidae) has been recognized as an aggressive invasive insect pest worldwide [P. solenopsis. These studies showed that P. solenopsis had strong temperature adaptability [Aenasius bambawalei Hayat (Hymenoptera: Encyrtidae) plays a significant role in controlling the mealybug population [P. solenopsis to many areas [P. solenopsis had recently invaded, the natural parasitism rate of A. bambawalei was significantly lower [A. bambawalei, and this result was even greater than that in its place of origin [A. bambawalei.For alien species, changes in development, morphology and fecundity at high temperatures are exhibited in various aspects ,12. The orldwide . In someorldwide . At presorldwide , developorldwide , and reporldwide  of P. sotability . Aenasiupulation ,20,21 anny areas . Howeverly lower . Our pref origin . Thus, wA. bambawalei at 27 °C and 36 °C. The results of this study will contribute to clarifying the adaptive mechanism to high temperature of this natural enemy and also further reveal the mechanism by which temperature changes affect the interaction between invasive insects and their exotic natural enemies.To verify this hypothesis, we compared the pupal development, hind tibia of female adults, and fecundity of P. solenopsis was field-collected from Hibiscus mutabilis grown in Hangzhou City, Zhejiang Province, China  in July 2012. In the laboratory, P. solenopsis was reared on cotton plants in a growth chamber  according to the procedures of our previous experiments [A. bambawalei were reared on their host P. solenopsis in the lab under the same controlled conditions as P. solenopsis. Third-instar nymphs and female adults of P. solenopsis were used in the experiments because of their relatively high survival rate and suitable duration of development. To maintain genetic stability and fitness of the population, A. bambawalei was collected from the wild and added to lab colonies irregularly.eriments . The colA. bambawalei, it was necessary to select a suitable temperature and instar that showed a significant change [A. bambawalei in this study. All wasp pupae  were divided into 2 groups: (1) pupae were collected, placed separately in 1.5-mL centrifuge tubes, and then maintained in the growth chamber at 27 °C until emergence  pupae were placed separately in a 1.5-mL centrifuge tube and heat-treated at 36 °C for 24 h in a climate-controlled incubator and then transferred to the climate room at 27 °C until emergence . Then, the emergence of A. bambawalei from these two groups was observed daily. The emergence time, number of parasitized P. solenopsis, and number of emerged A. bambawalei were recorded. Finally, as an important index to evaluate the size and fecundity of parasitoids [A. bambawalei was measured under a Nikon SMZ1500 zoom stereomicroscope . Each emergence time, emergence rate, and hind tibia length test included 60, 60, and 30 replicates, respectively.To evaluate the effect of temperature change on the development and reproductive traits of t change ,11. Accot change  and othet change , a 36 °CA. bambawalei were fed 10% honeydew solution, which was renewed every day until dissection. From 1 d after emergence, approximately 30 virgin females or virgin males per treatment group per day were dissected on average. The dissection was carried out according to the following steps: (1) the parasitoids were placed in the freezer  for approximately 2 min to freeze; and (2) the frozen parasitoids were immersed in PBS phosphate buffer solution and dissected under an anatomical microscope (Nikon SMZ-10) to detect intact or impaired female or male gonads. Following dissection, the ovaries per female or the testes per male were observed separately using a Canon D60 digital camera coupled to the microscope. The length of ovarioles was measured, and the ovariole length per female was calculated (mm). For ovariole length (mm), based on repeat measurements of a random subset of 30 individuals, the measurement error was very small. The numbers of oocyte stalk hook analogues and mature oocytes in the ovary were counted, and the mean number per ovariole was calculated. The diameter of all mature oocytes was measured, and each of the 3 largest mature oocytes was selected as the indicator (mm). These measurements were performed under a Nikon SMZ1500 zoom stereomicroscope  and used to determine the mean oocyte size per female.Newly emergent (within 12 h) A. bambawalei in incubators at 27 °C and 36 °C was up to 16 days; however, on the 12th day, the gonads began to degenerate. Thus, in this study, the data were analyzed at 12 days. Ovaries and testes in the control and high-temperature wasps were measured on the day of emergence, at 3 days old, at 5 days old, at 7 days old, and at 12 days old, and each treatment and control included 30 replicates. The criterion for male sterility was complete absence of sperm in the seminal vesicle or in the spermatheca of the mated female [As larger organs contain more sperm  and the d female .A. bambawalei in the number of parasitized P. solenopsis. The effect of high temperature on pupal development, hind tibia of female adults, and reproductive parameters was determined by one-way ANOVA with appropriate post hoc tests. Student’s t-test was used to compare the value of means from two samples, and the chi-square test was used to compare proportions.The emergence rate is the percentage of the number of emerged t-statistics with a Bonferroni correction. Mean values of female fecundity and egg viability were compared using a Mann-Whitney U test.Ovary and test dynamics were analyzed by a generalized linear model (GLM) using R statistical software . Mean values of oocyte number, egg, testis, and seminal vesicle size were compared using one-way ANOVA. Post hoc comparisons for all treatments were performed using a multiple comparison test based on Student’s F = 13.50, p < 0.05)  = 22.63, p < 0.05)  = 0.40, p = 0.54)  ( < 0.05) A and the < 0.05) C but had = 0.54) B. MoreovFemales had a pair of ovaries, and each ovary contained three ovarioles A–J. The In the early stages of emergence, there was a smaller ovarian volume. At the same time, the ends of the ovary were cemented into a group in an “S” shape and not easily separated A–C,K. WiThe mean ovary size increased with maternal age, and this increase was linear K. FemaleF = 29.89, p < 0.05). In any case, few or even no mature eggs were detected upon new emergence (n = 24.07) (n = 12.06)  D,Land on= 12.06) E,L. In a= 12.06) C–J. Then= 12.06) G–J. At t= 12.06) I,J,M.A. bambawalei is composed of a pair of testes (ts), vas deferens (vd), seminal vesicles (sv), accessory glands and a single ejaculatory duct (ed), and penis  was connected to the base of the testis, and the lower part of the vas deferens was enlarged into a yellow seminal vesicle (sv). On days 3 to 5 after emergence, the seminal vesicle was filled with mature sperm C–F.F = 194.74, p < 0.05)  = 1.1813, p = 0.278)  = 35.717, p < 0.05; width: F = 103.58, p < 0.05).High temperature had a strong impact on the development of testes I,J. Over < 0.05) I. Howeve= 0.278) J. OveralF = 69.239, p < 0.05; width: F = 29.735, p < 0.05). In heat-treated and control males, the seminal vesicle size increased with age and then decreased. On day 3, the seminal vesicle size under the 36 °C treatment reached the maximum value (0.126 × 0.071 mm), while the seminal vesicle size of the control peaked on day 4 (0.109 × 0.066 mm)  = 35.717, p < 0.05; width: F = 103.58, p < 0.05).At all ages, males at high temperature had larger seminal vesicles than control males K,L . P. solenopsis originated in North America and then invaded many countries, including those in South America, Africa, and Asia [P. solenopsis shows that there has been great differentiation between the two groups [A. bambawalei has changed in different regions to speculate whether this parasitoid is adapting to global warming.At present, compared with more reports focusing on the impact of high temperature on the distribution ,13,23,32o groups . At the o groups . TherefoA. bambawalei according to other studies. Unexpectedly, some oocyte stalk hook analogues were produced during egg maturation. After the egg disappeared, this structure persisted in the ovary. A previous study demonstrated that oocytes remain attached to the ovary by a cellular pedicel until the oocytes are released into the ovary lumen [A. bambawalei and renewed it every day until dissection. This nutrient source has been proven to provide parasitoids with the necessary nutrients [To determine the egg quantity more accurately, we deprived the host of ry lumen . Howeverry lumen . This isry lumen  or the rry lumen . In thisutrients . MoreoveA. bambawalei develop into females, while unfertilized eggs develop into males [A. bambawalei. In contrast, high temperature may cause delays in spermatogenesis [Nasonia vitripennis, the same heat treatment was proven to be harmful to sperm quantity but did not impair sperm quality [Finally, similar to other hymenopteran insects, the fertilized eggs of to males . For parto males . Thus, ito males . Parasitto males . Thus, sto males . In a feto males . In the ogenesis  or decreogenesis , and is  quality . To moreA. bambawalei.Since global warming does not produce an even impact on all ecological habitats, how global warming will affect parasitoids and their role as natural enemies of pests is difficult to evaluate in the short term. However, the results of this study are still helpful for understanding the adaptive mechanisms of A. bambawalei. Therefore, this may explain, to some extent, why A. bambawalei can migrate with the mealybug, and why high temperature can increase the sex ratio of the offspring of A. bambawalei. Future studies should be carried out on the reproductive physiology, biochemistry, and molecules involved to clarify the specific accompanying mechanism. A clear definition of this mechanism will be conducive to the better application of A. bambawalei to prevent and control P. solenopsis and will enable the prevention and control of other biological invasions under the current situation of high temperature.A study of the developmental, morphological, and fecundity changes in the parasitoid at high temperature therefore provides crucial insight into its response to temperature change ,46. In t


Quality of Evidence in Studies estimating Prevalence of Exposure to Occupational risk factors (QoE-SPEO) approach. We describe QoE-SPEO’s development to date, demonstrate its feasibility reporting results from pilot testing and case studies, note its strengths and limitations, and suggest how QoE-SPEO should be tested and developed further.The World Health Organization (WHO) and the International Labour Organization (ILO) have produced the WHO/ILO Joint Estimates of the Work-related Burden of Disease and Injury (WHO/ILO Joint Estimates). For these, systematic reviews of studies estimating the prevalence of exposure to selected occupational risk factors have been conducted to provide input data for estimations of the number of exposed workers. A critical part of systematic review methodology is to assess the quality of evidence across studies. In this article, we present the approach applied in these WHO/ILO systematic reviews for performing such assessments on studies of prevalence of exposure. It is called the iP) between individual raters and rater teams for the downgrade domains. iP ranged between 0.00 (no two pilot testers selected the same rating) and 1.00 . Case studies were conducted of experiences of QoE-SPEO’s use in two WHO/ILO systematic reviews.Following a comprehensive literature review, and using expert opinion, selected existing quality of evidence assessment approaches used in environmental and occupational health were reviewed and analysed for their relevance to prevalence studies. Relevant steps and components from the existing approaches were adopted or adapted for QoE-SPEO. New steps and components were developed. We elicited feedback from other systematic review methodologists and exposure scientists and reached consensus on the QoE-SPEO approach. Ten individual experts pilot-tested QoE-SPEO. To assess inter-rater agreement, we counted ratings of expected  heterogeneity and quality of evidence and calculated a raw measure of agreement (effect of exposures. Assessments using QoE-SPEO comprise three steps: (1) judge the level of expected heterogeneity , (2) assess downgrade domains, and (3) reach a final rating on the quality of evidence. Assessments are conducted using the same five downgrade domains as the Grading of Recommendations Assessment, Development and Evaluation (GRADE) approach: (a) risk of bias, (b) indirectness, (c) inconsistency, (d) imprecision, and (e) publication bias. For downgrade domains (c) and (d), the assessment varies depending on the level of expected heterogeneity. There are no upgrade domains. The QoE-SPEO’s ratings are “very low”, “low”, “moderate”, and “high”. To arrive at a final decision on the overall quality of evidence, the assessor starts at “high” quality of evidence and for each domain downgrades by one or two levels for serious concerns or very serious concerns, respectively. In pilot tests, there was reasonable agreement in ratings for expected heterogeneity; 70% of raters selected the same rating. Inter-rater agreement ranged considerably between downgrade domains, both for individual rater pairs (range iP: 0.36–1.00) and rater teams (0.20–1.00). Sparse data prevented rigorous assessment of inter-rater agreement in quality of evidence ratings.We found no existing quality of evidence assessment approach for occupational exposure prevalence studies. We identified three relevant, existing approaches for environmental and occupational health studies of the We present QoE-SPEO as an approach for assessing quality of evidence in prevalence studies of exposure to occupational risk factors. It has been developed to its current version (as presented here), has undergone pilot testing, and was applied in the systematic reviews for the WHO/ILO Joint Estimates. While the approach requires further testing and development, it makes steps towards filling an identified gap, and progress made so far can be used to inform future work in this area. These oTo estimate the burden of disease attributable to exposure to a risk factor, it is crucial to estimate how widespread  exposure to the risk factor is. Five of the WHO/ILO Joint Estimates systematic reviews aimed to synthesise studies estimating prevalence of exposure  to occupational risk factors: ergonomic risk factors, dusts and/or fibres, solar ultraviolet radiation, noise, and long working hours . An examFramework for the Use of Systematic Review in Chemical Risk Assessment (The need for systematic reviews of studies estimating exposure to risk factors is increasingly being recognized (beyond the WHO/ILO Joint Estimates). Through its sessment  WHO has sessment , includisessment .•Risk of Bias in Studies estimating Prevalence of Exposure to Occupational risk factors (RoB-SPEO) tool (the EO) tool ).•Quality of Evidence in Studies estimating Prevalence of Exposure to Occupational risk factors (QoE-SPEO) approach (presented in this article).the Despite this recognition of the need for systematic reviews of exposure prevalence studies, to our World Health Organization 2021b knowledge there are no standardized methods for such reviews. Thus, to address this identified gap, WHO and ILO, supported by individual experts, have developed:WHO and ILO have sought to ensure that RoB-SPEO and QoE-SPEO are compatible and complementary.1.1Occupational exposure prevalence studies determine the presence (and often the level or intensity) of an exposure of interest to an occupational risk factor of interest in each individual in the study population or in a representative sample at one particular time point . The preStudies estimating prevalence are distinct from studies estimating incidence  or studExposure is the “proximity and/or contact with a source of a disease agent (or hazard) in such a manner that effective transmission of the agent or harmful effects of the agent may occur” . There aExposures to occupational risk factors are biological, chemical, physical, ergonomic, mechanical and psychosocial exposures among workers at their workplace, posing a risk known to be harmful to human health . One exaOne key and new concept for systematic review of studies estimating prevalence of exposure to occupational risk factors is “expected heterogeneity”  . Quality1.2.1studies of the effect of exposure to an environmental or occupational risk factor on a health outcome . Consequently, prevalence studies investigate neither health outcomes, nor effects, and therefore, methods in existing approaches for assessing related quality of evidence are not applicable.1.2.1.2the effect of an exposure on a health outcome may come from evidence streams other than just human data . Conversely, evidence on the prevalence of (human) exposure to an occupational risk factor comes exclusively from data based on human studies. Therefore, while methods for assessing quality of evidence in studies of health effects of exposure to occupational risk factors need to be able to assess quality of evidence across evidence streams, methods for assessing quality of evidence in prevalence studies need to consider human-based data only.Evidence regarding 1.2.1.32I (quantifying the proportion of variation in relative risks due to among-study differences) is large , which includes improvements based on user feedback after it was applied (in its third version) in the WHO/ILO Joint Estimates, as the approach applied for assessing quality of evidence in prevalence studies of exposure to occupational risk factors in the series of WHO/ILO systematic reviews . Our des2WHO and ILO developed QoE-SPEO with the support of a WHO/ILO Working Group of individual experts on systematic review, environmental and occupational health, and exposure science. Currently QoE-SPEO has been developed up to v.4; the steps taken so far in its development process are summarized in 2.1In June 2018, we undertook a comprehensive, but non-systematic, purposive review of the literature in the electronic bibliographic databases Ovid Medline and EMBASE, to identify and select existing quality of evidence approaches that might be relevant to assessing prevalence studies of exposure to occupational risk factors.•focusing on prevalence studies, incidence studies, prognostic studies, studies on the effect of exposure to occupational risk factors on health outcomes, and/or studies of the effect of occupational health or safety interventions on health outcomes;•for assessing the quality of evidence at the level of the entire body of evidence;•for assessing quality of evidence in non-randomized studies of human data, because pertinent prevalence studies for our exposure systematic reviews are of this type; or•based on checklists and/or quantitative scoring (conscious that scoring is not recommended in GRADE or Cochrane due to inherent biases and judgements in scoring).We included approaches:•for assessing quality of single studies;•specifically designed for randomized study designs, and in vitro, in vivo and/or mechanistic evidence streams; or•designed for purposes other than assessing quality of the body of evidence (such as risk of bias tools and reporting guidelines).We excluded approaches:Thirty experts in systematic review methods, environmental health, occupational health, occupational safety, and exposure science were also asked to identify relevant approaches. Each expert was a contributor to one or more systematic reviews for the development of the WHO/ILO Joint Estimates .2.2•non-randomized studies;•studies estimating any type of prevalence  with expected heterogeneity judged as “high” or “moderate” and (ii) with a “low” level of expected heterogeneity.We also identified gaps in the current approaches and developed new components that were key for QoE-SPEO. We created all three components that formed the new step for assessing the level of expected heterogeneity in the prevalence. We also developed a reporting table for the new component of recording judgments, ratings and/or rationales for grading evidence down. At the end of this process, we had developed an initial prototype approach (QoE-SPEO version v.1).2.4We sought and integrated feedback on QoE-SPEO in three stages , from a 2.5After receiving initial feedback on QoE-SPEO v.1 from seven experts and implementing changes, we undertook pilot-testing QoE-SPEO v.2. One aim of this first round of testing was to receive more feedback to further develop the approach. Another aim was to assess inter-rater agreement . There is minimal scientific consensus on which of several existing methods is preferable for calculating inter-rater agreement. Inter-rater agreement ratings are also not necessarily indicators of an approach’s performance, since ratings are explicitly based on individual judgment; therefore, transparency and reaching a consensus are more important . We neve•all study records, unpublished analyses and/or re-analyses of the included studies;•table of characteristics  of the i•tables of risk of bias  for the •figure of summary of risk of bias ; and•pooled prevalence estimates produced using the input data collected in the systematic reviews ;3.record the outcomes  of the individual and consolidated assessments in a specific Excel recording sheet; and4.propose revisions to address issues identified and record issues that the team was unable to address.Second, ten co-authors of this article (none of whom had previously contributed to QoE-SPEO’s development) pilot tested QoE-SPEO v.2. We listed pilot testers alphabetically by surname and then paired consecutive pilot testers. They were asked to:1.Agreement on ratings of expected heterogeneity between individual raters in QoE-SPEO Step 1: We calculated the proportion of pilot testers who rated the level of expected heterogeneity as: (1) “no or only minor”; (2) “low”; (3) “medium”; and (4) “high”.2.j-th analytical category, using the following formula:Agreement on ratings of downgrade domains between individual raters in QoE-SPEO Step 2: We assessed agreement for ratings of QoE-SPEO’s five downgrade domains between the ten pilot testers, using the individual assessments. Ratings were coded into three analytical categories: (1) “no or only minor concerns”, (2) “serious concerns”, and (3) “very serious concerns”. Using established methods , we calcThird, we preliminarily tested agreement in ratings between pilot testers by step and/or component, using the following metrics:i = 1,…k is the number of domains ; j = 1,…k is the number of possible analytical categories ; and n = number of assessors for the component (here n = 10). iP ranges from 0.00 (no two pilot testers chose the same rating) to 1.00 .3.Agreement on domains between rater teams in QoE-SPEO Step 3: We replicated the methods used to calculate agreement on ratings of downgrade domains between individual raters (see 2. above) to calculate such agreement between rater teams, using the teams’ consolidated assessments.4.Agreement on ratings of quality of evidence in QoE-SPEO Step 3: We counted the number of pilot testers who rated the quality of evidence as (1) “very low”; (2) “low”; (3) “moderate”; and (4) “high”.where Finally, we integrated all feedback received from pilot testers, including proposals for revisions to QoE-SPEO v.2, and reports of issues, to develop QoE-SPEO v.3. An additional round of testing of QoE-SPEO v.3 was carried out opportunistically, as feedback on the tool was sought from 20 individual experts in occupational health, occupational safety, and/or exposure science after they applied it in a systematic review of prevalence studies for the WHO/ILO Joint Estimates. Three improvements were made following this round of testing: we added further explanation regarding the concept of expected heterogeneity; we refined considerations and criteria for some downgrade domains; and we added definitions of quality of evidence ratings to provide more information for raters as this was the downgrade domain with poor inter-rater agreement.2.6We conducted two case studies to collect and report reflections from users that applied QoE-SPEO v.4 in WHO/ILO systematic reviews for use in the further development and testing of the approach. All quality of evidence assessors (N = 5) from two of the systematic reviews of the WHO/ILO Joint Estimates  were sen33.1No existing approach was identified for prevalence studies (at least not for assessing a body of evidence). We excluded several approaches from our review e.g., . These wWe considered three existing approaches for assessing quality of evidence to be most relevant : the GRA3.2a priori the baseline level at which the quality of evidence assessment commences based on features of study design). We identified two of these four steps to be relevant for QoE-SPEO: (i) assess downgrade domains and (ii) reach a final decision (rating) on the quality of evidence. We, therefore, adopted these and adapted them as necessary  is applicable only to studies of effect estimates, and the underlying concepts of large effect size, dose–response relationship and confounding do not transfer to prevalence studies.We added one step in QoE-SPEO not used in the three existing approaches : assessm3.3Of the 12 components identified from the three existing approaches, we selected seven for inclusion in QoE-SPEO Steps 2 and 3 . As StepWe also developed four new components for QoE-SPEO. Three of these were for Step 1. The other was the introduction of reporting tables in Steps 2 and 3.3.4The QoE-SPEO approach, as developed up to v.4, is presented in full in Appendix A of the Supplementary data. This is the approach that WHO and ILO, supported by a large number of individual experts, applied in their series of systematic reviews to assess the quality of evidence in occupational exposure prevalence studies for the WHO/ILO Joint Estimates .3.4.1The QoE-SPEO approach provides general instructions for assessors. These introduce the assessor to the aims, overall structure (including steps and components) and standard format of the approach. Importantly, the assessor is instructed to always record their selected rating (or decision) and detailed justification for it for each step, using the relevant recording table provided at the end of each section.3.4.2•Step 1: Judge the level of expected heterogeneity.•Step 2: Assess downgrade domains.•Step 3: Reach a final decision (rating) on the quality of evidence.In QoE-SPEO, assessors consecutively undergo the following three steps:Each step comprises upfront definitions of key concepts related to that step , instructions specific to the step, considerations for assessments, standard ratings, criteria for ratings (if applicable), and a table for recording and documenting the selected rating and the rationale. Some instructions and considerations, especially regarding downgrading in some domains, differ depending on the level of expected heterogeneity.I2 statistic  heterogeneous, a body of evidence should not be downgraded if it is found to be  heterogeneous for prevalence estimates. When prevalence estimates have been pooled, one indicator of high statistical heterogeneity can be a high tatistic . Step 1 1.Risk of bias.2.Indirectness.3.Inconsistency.4.Imprecision.5.Publication bias.Step 2 involves assessors considering and rating the five downgrade domains:This step is also completed independently for each outcome . For each downgrade domain, the assessor can downgrade the quality of evidence for “serious” concern by one level (-1) or “very serious” concern by two levels (-2). The ratings and justification for each downgrade domain are recorded in the tables for this purpose.In Step 3, each assessor grades the quality of evidence as “high quality”, “moderate quality”, “low quality” or “very low quality”. This step consolidates any downgrading from Step 2, with consensus reached across a rater team. Grading always starts at “high quality” of evidence, with downgrading as appropriate. The final rating with its justification is recorded to ensure transparency.3.5We now present the results from the pilot testing of QoE-SPEO v.3.3.5.1The level of expected heterogeneity was rated by three of the ten individual raters (30%) as “medium” and by seven (70%) as “high”. None rated the level as “none or only minor” or “low”. We judged this to indicate substantial inter-rater agreement. However, this examination of agreement is simple and relies on a small number of raters. Additional testing is needed to establish inter-rater agreement for this step with more certainty.Agreement between individual raters in ratings of downgrade domains in Step 2 is shown in Low levels of agreement for the Risk of bias domain can be at least partially explained by the fact that this domain is particularly complex to analyse (e.g. risk of bias assessments using RoB-SPEO comprise consideration of eight different types of bias).3.5.2Agreement between rater teams in ratings of downgrade domains from Step 3 is also displayed in 3.5.3The quality of evidence was rated as “high” by one of the five rater teams (20%), “moderate” by one team (20%) and “low” by three teams (60%). None of the rater teams rated it as “very low”. These data are too sparse to rigorously analyse. However, the ratings of both “low” and “high” were each ascertained by at least one group. Additional tests with larger counts of ratings are needed.44.1The QoE-SPEO approach that is presented here and was applied in the systematic reviews for the WHO/ILO Joint Estimates differs considerably from other quality of evidence assessment approaches in environmental and occupational health. a)The QoE-SPEO approach assesses the quality of evidence across studies estimating the prevalence of exposure to occupational risk factors, whereas the other existing approaches assess quality of evidence across studies estimating intervention effectiveness, causal effects on (or association with) health outcomes of environmental and occupational risk factors.b)QoE-SPEO assesses the quality of evidence along domains relevant to non-randomized prevalence studies within the human evidence stream only. The other three tools, on the other hand, facilitate assessment along domains relevant to studies using various evidence streams . However, they can be tailored to assess the human evidence stream only, as in several of the systematic reviews for the WHO/ILO Joint Estimates, including .c)QoE-SPEO includes a step with relevant components that involve judging expected heterogeneity of the prevalence of interest. Expected heterogeneity is a novel concept in quality assessment that is likely to be particularly relevant for assessing exposure prevalence studies. The assessment of some downgrade domains, especially inconsistency and imprecision, need to be considered in light of the level of expected heterogeneity.d)QoE-SPEO always starts assessments with the rating of “high quality” of evidence, as all study designs have the potential to provide high quality evidence on exposure prevalence to occupational risk factors (at least in principle). The other three approaches determine the baseline level of quality of evidence based on study design.e)While all four approaches assess the same five downgrade domains, the specific criteria and considerations for downgrading are necessarily quite different between QoE-SPEO and the three existing approaches.f)While the existing three approaches assess upgrade domains, QoE-SPEO does not, because the upgrade domains of existing approaches are not applicable to prevalence studies.g)QoE-SPEO adopts the four ratings of quality (or confidence or certainty) of evidence used in GRADE and OHAT, rather than the three Navigation Guide ratings. For rating prevalence studies, we preferred a more granular set of ratings. Our approach minimally modifies the standard explanation for GRADE ratings to suit assessments of evidence on occupational exposure prevalence .The main differences between QoE-SPEO and the other approaches are:4.24.2.1Strengths of QoE-SPEO include its rigorous development through iterative rounds of feedback from diverse experts on systematic review methods, occupational health and safety, environmental health and, importantly, exposure science. The approach was developed jointly by systematic review methodologists and practitioners who apply evidence synthesis in policy, ensuring that the proposed approach is applicable not only in theory but also feasible in policy practice, including at the global level. QoE-SPEO builds on, and is aligned with, the GRADE framework and also draws on other relevant approaches. It is designed to be used in combination with the RoB-SPEO risk of bias tool and adhere to the GATHER reporting guidelines . The use4.2.2QoE-SPEO is the first, and currently the only, domain-based approach for assessing quality of a body of prevalence studies. It introduces methodological innovations, including the concept of “expected heterogeneity” of the prevalence. While expected heterogeneity may not be a consideration for other types of studies and therefore not a component in existing approaches, it is key in studies of prevalence of exposure, influencing the assessment of downgrade domains and quality of evidence. A large number of geographically diverse individual experts working within and across different disciplines  has pilot tested and applied QoE-SPEO in systematic reviews for the development of official, global health norms . The preliminary results from these tests and real-world applications suggest that QoE-SPEO performs well across most steps and components within steps, achieving an overall good to very good level of inter-rater agreement for almost all domains. Users of the approach have reported to the principal author of this article that QoE-SPEO was comparatively easy and time-efficient to use, with some exceptions; reflections from users are shown below in 4.2.3Our review of existing approaches for assessing the quality of a body of evidence was not comprehensive. However, we asked several experts to identify existing approaches for assessing quality of evidence in occupational exposure prevalence studies, and as none were identified, we are confident in our conclusion that no such approach currently exists. Assessors may not be familiar with the novel concept of “expected heterogeneity”, but we have tried to mitigate this by explicitly adding its definition in QoE-SPEO. Testing of QoE-SPEO has been limited so far. Feedback was received from 20 individual experts who trialled QoE-SPEO v.3 (not the improved QoE-SPEO v.4 presented in this article). This involved assessment of one specific body of prevalence studies only , and comprised a relatively small number of raters and rater teams. The data were thus too sparse for robust analysis and conclusions. Inter-rater agreement was poor for the downgrade domain of risk of bias.We recognise that this approach requires further reflection and development. It should be considered that the reflections from applying the approach shown in 4.3Box 1Box 2Assessors from two of the systematic reviews of the WHO/ILO Joint Estimates provided their reflections on applying the QoE-SPEO approach. 4.4Additional performance testing of QoE-SPEO is required to improve and develop the tool further. This should include a comprehensive assessment of inter-rater agreement for ratings of expected heterogeneity, downgrade domains and quality of evidence. Indicators of inter-rater agreement by themselves cannot establish epistemological reliability  . Moreove•Step 1  is novel and should, therefore, be reviewed and revised as needed. As it should be carried out as a first step, and to conduct this assessment before assessors have knowledge of the evidence base, it may be most appropriate to move this step to the protocol stage of a systematic review and report the assessment in the protocol before the systematic review commences. Assessors may benefit from further examples and guidance regarding the assessment of expected heterogeneity and how this might influence decisions for the downgrade domains. A set of criteria to evaluate this new concept should be developed. Also, this step may need substantial further refinement following more definitional and conceptual research, such as move to a matrix-type assessment of expected heterogeneity by geographic region, industrial sector, sex and/or other determinants of heterogeneity.•In Step 2, the downgrade domain of risk of bias may require changes to improve inter-rater agreement. For example, regarding the domain of imprecision, since occupational prevalence estimates from single studies and from meta-analyses commonly have highly skewed 95% confidence intervals , more detailed guidance could be developed to judge imprecision in prevalence estimates with skewed confidence intervals.•QoE-SPEO’s scope may include mathematical or statistical modelling studies  and studies that use biomarkers for measuring exposure. Dedicated steps and components may need to be developed to assess the quality of evidence in these studies.•point prevalence . While this was based on the judgement that theoretically studies of all designs could be used to measure a prevalent exposure, some designs may be less suited to a specific research question. For example, cross-sectional studies are appropriate for measuring evalence  of an ex•Evidence might emerge that will support the use of upgrade (or other downgrade) domains.We have identified some areas for consideration when the QoE-SPEO approach is developed further:QoE-SPEO will benefit from further development and refinement over time. We welcome comments from those with expertise on the topic and invite organizations and individual experts to test the approach and provide feedback. This will help further tailor and refine tool components. We recognise, however, that the gaps for systematic reviews of studies of prevalence of exposure to occupational risk factors are not limited to the need for an approach to assess quality of evidence. A wider framework is required to help systematic reviewers develop research questions, and assess risk of bias and quality of evidence.5QoE-SPEO was applied in its third version to assess the quality of bodies of evidence from occupational exposure prevalence studies in a series of harmonized systematic review and meta-analsyes conducted by WHO and ILO, supported by individual experts, for the development of the WHO/ILO Joint Estimates of the Work-related Burden of Disease and Injury (WHO/ILO Joint Estimates). After amending QoE-SPEO based on feedback from the users, we have developed it, up to its fourth iteration, as described in this paper. We propose this as an approach for assessing quality of evidence in prevalence studies of exposure to occupational risk factors and present the considerations made while developing this tool. QoE-SPEO will benefit from further testing and development, particularly also by assessors from other disciplines and backgrounds, but could be applied in its current form in systematic reviews for health risk assessment at the exposure assessment step, in guideline and policy development, and for the production of health estimates. The approach could also be considered for assessment of quality of evidence in prevalence studies of exposure to other human health risk factors . Applications in other fields need careful consideration and modifications may be needed.All authors are salaried staff members of their respective institutions. This publication was prepared with financial support to the WHO from the National Institute for Occupational Safety and Health of the Centres for Disease Control and Prevention of the United States of America ; the German Federal Ministry of Health (BMG Germany) under the BMG-WHO Collaboration Programme 2020–2023 (WHO specified award ref. 70672); the Spanish Agency for International Cooperation (AECID) (WHO specified award ref.71208). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.Environment International; Lancaster Environment Centre, Lancaster University, United Kingdom)Conceived the idea: FP, Paul Whaley (Associate Editor for Systematic Reviews, Searched literature and provided expert opinions on approach selection: FP, DG, RLMDeveloped first prototype of approach: FP, DGLed all aspects of approach development: FPContributed to approach development: DG, LB, DM, RLM, SLNDeveloped methodology and facilitated pilot testing: FPPilot tested the approach: NC, AD, FB, LG, TL, AM, DP, MSP, PS, TTAnalysed data collected in pilot tests: FPProvided reflections on user experience: HFvM, AMD, KCP, SN, LTWrote first draft and led development of article: FPContributed to manuscript writing and development: All authorsRevised the manuscript based on comments from co-authors: FP, NCM8The authors alone are responsible for the views expressed in this article and they do not necessarily represent the views, decisions or policies of the institutions with which they are affiliated.The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper.


This project describes the creation of a single searchable resource during the pandemic, called the COVID-19 Best Evidence Front Door, with a primary goal of providing direct access to high-quality meta-analyses, literature syntheses, and clinical guidelines from a variety of trusted sources. The Front Door makes relevant evidence findable and accessible with a single search to aggregated evidence-based resources, optimizing time, discovery, and improved access to quality scientific evidence while reducing the burden of frontline health care providers and other knowledge-seekers in needing to separately identify, locate, and explore multiple websites. Early on in the COVID-19 pandemic, a need for evidence-based information was recognized, and as a partnership endeavor, the Taubman Health Sciences Library and the Department of Learning Health Sciences at the University of Michigan worked together to create a searchable database of relevant resources for frontline health care professionals. A team of informationists, faculty, researchers, and administrators developed a rapid prototype resource, and then a group of volunteers collaborated with the team to identify and review documents for inclusion.This effort resulted in the creation of a single searchable resource, called the COVID-19 Best Evidence Front Door, providing direct access to high-quality meta-analyses, literature syntheses, and clinical guidelines from a variety of trusted sources. The Front Door makes relevant evidence findable and accessible with a single search, reducing the search time for frontline health care providers and other knowledge-seekers in locating high-quality, evidence-based COVID-19 information resources . The datTo determine which documents to include in the database, based on the best available evidence, a draft set of criteria was developed and then rapid pilot tested by five project team members who applied the criteria to guideline searches . FollowiTo organize documents for review and to manage the workflow, two online Google spreadsheets were created and maintained as the “document base.” The first spreadsheet is fluid and ever-changing and is used by project team members, supplemented by a group of interested volunteers, as a working list of potential articles for review with a core set of information provided about each guideline, including complete title, dates of publication, working hyperlink, and keywords. The second spreadsheet is used to capture and manage static, moment-in-time snapshots of the document base. These snapshots are accessible and searchable online.The Front Door database has been developed using Knack and Vee Technologies. Knack is a platform for creating a combined website and searchable database without requiring a lot of coding expertise. The interface is easy to use and intuitive, making data quickly and securely searchable and sharable online. Librarians and other information providers can use Knack as a customizable tool to expedite the creation of a website that is more than just descriptive text and hyperlinks to other resources because it also enables the organizing, structuring, and searching of data from existing spreadsheets.Updating the database becomes an efficient process as well. The Front Door prototype website and database were created and ready for user testing within an eight-day period, and once developed, the updating of the database is now a six-step process, taking approximately thirty minutes of time. The database accepts CSV file uploads, which enables easy integration into the Front Door project team's Google Sheets workflow and uploading of document base snapshots. A multipart search capability was also deployed, allowing users to search the document base by keyword, publication date, source organization, and document type. Knack provided their fee-based services and software free of charge for ninety days because of the primary focus on providing access to COVID-19 resources during the pandemic. Since the building and testing of the resource was a concentrated effort, requiring only slightly more than a week of time, the Front Door's availability was able to be announced in April 2020 within a month of COVID-19 surging and a concerted move to socially distanced and remote work.A similar partnership with Vee Technologies enhanced the ability to more rapidly keep up with fast-changing content about COVID-19. Vee Technologies is a company that provides bots that automatically search the web for information on specified resources. This automation is an efficiency tool, enabling quicker identification of resources of interest and improving data collection. For the Front Door project, this technology featured an automation bot to scrape COVID-19 documents from selected source organizations' websites. The company sent emails, twice per week, with updated lists of new COVID-19 document metadata from the selected websites for review for possible inclusion in the Front Door. This service was provided free of charge, with only a request for acknowledgment on the Front Door website, and it enabled quicker identification of potential COVID-19 documents for review and inclusion.http://covid19bestevidence.org/>. The site currently provides searchable access to over 350 documents from over 60 different sources . Google Analytics was added to the Front Door prior to launch to capture access and usage data. The database has been utilized by over 3,400 visitors to the site between April 21, 2020, and May 27, 2021, with a total of 4,383 sessions (“the period of time a user is actively engaged with [the] website”) [The Front Door is publicly available at <New features continue to be explored, including interactive visual document filtering to further increase ease-of-use and user engagement. The interactive pie charts present users with a method for grouping and filtering documents based on document criteria and search terms of interest . Once thContent continues to be updated as well, including documents on recent vaccine-related evidence as they become available. To increase ease of accessibility for this information, a “Vaccine” tab has been added, which includes links to vaccine articles generated by four PubMed queries. Two of the four queries search title and title/abstract of FDA-authorized COVID-19 vaccine information, and the remaining two queries search title and title/abstract of COVID-19 vaccine information in general.Visitors to the site can also sign up for weekly updates which include new evidence documents as well as preexisting documents that have been revised.


Low back pain is a leading cause of disability worldwide. Health literacy has been associated with pain intensity and pain control. However, there is a paucity of evidence regarding this association. In the field of low back pain research, inconsistent reporting of outcomes has been highlighted. To address this issue a Core Outcome Set has been developed.The objectives of this scoping review were: (1) The health literacy measures currently employed for low back pain and the aspects of health literacy they include. (2) The low back pain health outcomes included in such work. (3) The extent to which these health outcomes reflect the Core Outcome Set for Clinical Trials in Non-Specific Low Back Pain.The search included thirteen bibliographic databases, using medical subject heading terms for low back pain and health literacy, and followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews guidelines. The eligibility criteria were defined by the Joanna Briggs Institute PCC mnemonic. A thematic framework approach was used for analysis.The search yielded ten relevant studies for inclusion, amongst which a total of nine health literacy measures and 50 health outcome measures were used. Most health literacy measures focused on functional health literacy, with few assessing communicative and critical health literacy. The health outcomes assessed by the included studies could be broadly categorised into: Pain, Disability, Behaviour, Knowledge and Beliefs, and Resource Utilisation. Most of these outcome measures studied (36 out of 50) did not directly reflect the Core Outcome Set for Clinical Trials in Non-Specific Low Back Pain.To allow for comparison across findings and the development of a rigorous evidence base, future work should include the Core Outcome Set for Clinical Trials in Non-Specific Low Back Pain. There is an urgent need to broaden the evidence-base to include regions where low back pain morbidity is high, but data is lacking. Such work demands the incorporation of comprehensive measures of health literacy that have both generic and culturally sensitive components. Low back pain (LBP) is the single leading cause of disability globally and is rising , 2. In 2Effective self-management is crucial to improving LBP outcomes , 10. StuDespite the need for a stronger evidence base in LBP management, inconsistent reporting of outcomes in clinical trials of patients with LBP has been highlighted . This poThere also exists a paucity of research to underpin evidence-based practice of LBP treatment in low- and middle-income countries (LMIC) , 24. ThiTo develop more evidence-based interventions and guidelines we need to better understand the relationship between HL and LBP outcomes. An initial scoping search of the literature was conducted to assess whether reviews and guidelines on this topic have already been published and what was lacking. This only yielded a single systematic review by Edward et al. in 2018 on the impact of HL on LBP management. The study identified only three relevant studies, all of which were based in high income Western nations. However, the authors of the review acknowledged “possible incomplete retrieval of identified research and reporting bias”  as the sThe health literacy measures currently employed for low back pain and the aspects of health literacy they include.The low back pain health outcomes included in such work.The extent to which these health outcomes reflect the Core Outcome Set for Clinical Trials in Non-Specific Low Back Pain. Scoping reviews are used instead of systematic reviews where the purpose of the review is to identify knowledge gaps, scope a body of literature, clarify concepts or to investigate research conduct [This scoping review builds on Edward et al.’s work and had three objectives. These were to methodically map evidence on: conduct . This me conduct . To do s conduct , this st, Web of Science, Cochrane, Google Scholar, and ClinicalKey.The searches were conducted in: MEDLINE, Pubmed, Academic Search Complete, The Cumulative Index to Nursing and Allied Health Literature, Education Source, Education Resource Information Centre, PsycINFO, Global Health, Embase (Ovid)Back Pain, Back Ache, Back Pain with Radiation, Back Pain without Radiation, Backache, Vertebrogenic Pain Syndrome, Low Back Pain, Low Back Ache, Low Back Pain Mechanical, Low Back Pain Posterior Compartment, Low Back Pain Postural, Low Back Pain Recurrent, Low Backache, Lower Back Pain, Lumbago, Mechanical Low Back Pain, Postural Low Back Pain, Recurrent Low Back Pain.MeSH  terms used included: Health Literacy. No additional search filters were applied. See “Appendix The MeSH term used to search for HL was The search strategy was developed in consultation with the library team at the University, as well as expert opinion within the research team, which consisted of a range of expert researchers and clinicians . This inThe Joanna Briggs Institute (JBI) manual’s PCC mnemonic  was usedPopulation—Patients with LBP (≥ 10% of study population), of any age, gender, or raceConcept—Relationship of LBP health outcomes to HLContext—Any healthcare setting, in any geographical settingOnly research studies were included in this scoping review as the objectives of this study focused on measures used in LBP research. Hence other sources of evidence  were not included. In addition, studies for inclusion required the use of specific HL and health outcome measures. Studies were excluded if they only analysed generic literacy, numeracy, and education level not in the context of healthcare. Generic patient education interventions have the potential to influence non-HL related determinants of LBP, hence drawing conclusions about HL’s effects on LBP from these studies may be inaccurate [accurate , and forThe search strategy followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines . An indeData extraction included determinants of HL  , study dAs this was a scoping review, grading of evidence was not conducted. Instead, this study followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses extension for Scoping Reviews (PRISMA-ScR) guidelines , as detaThe initial search yielded 5509 articles. After removing duplicates and reviewing titles and abstracts, 18 articles remained for full-text review. Ten of these were included in the final analysis  and had a majority female population, with one study having entirely female participants. Racial break-down was only provided by two USA studies, both of which had predominantly white study populations. Nine studies collected data on education level, most reporting an even spread across participants.Nine different HL measures were used across the ten studies Table . Most HLIn their 2010 paper, Briggs et al.  and 0.97Subsequently in their 2011 paper, Briggs et al. used the Health Literacy Measurement Scale (HeLMS) , a psychThe Newest Vital Sign (NVS) was the most frequently used HL measure Table , utiliseFarin, Ullrich and Nagl developed the HELP questionnaire , an 18-item assessment that aims to summarise a patient’s reported communication and comprehension difficulties in health education and treatment . The queCamerini and Schulz  interpreMeasurement of functional HL alone is also seen in other studies. Burke, Nahin and Stussman used the response option “Never heard of it/Do not know much about it” from the National Health Interview Survey, arguing that this serves as an indicator of health knowledge which in turn is a correlate of functional HL . MacLeodKöppen et al. used HL questions taken from the Brief Questions to Identify Patients with Inadequate Health Literacy , a screeIn addition to the NVS mentioned above, Glassman et al. also used The Health Literacy Assessment, a 10-item self-administered questionnaire using items selected from the computerized Health LiTT measure . The HeaHL was associated with a wide range of outcomes Table . Five ovPainDisabilityBehaviourKnowledge and BeliefsResource UtilisationSeven studies involved data on pain , 48, 51,Five studies involved data on disability , 48, 51.Five studies collected data on patient behaviours , 51, 53,Four studies gathered data on patient knowledge and beliefs –17, 25, Four studies involved data on resource utilisation , 53, 54.A total of 50 health outcome measures were utilised across the ten studies reviewed. Of these, 14 (28%) were deemed to be directly related to those in the COS but were limited to two outcomes “pain intensity’ and “physical functioning”  Table 66. The PaAlthough not a primary aim of this scoping review, we briefly detail here findings on the association between HL and LBP health outcomes as a snapshot of existing literature Table . Out of We will now discuss our results in the context of the three objectives and the implications for evidence and future research i.e. (1) The health literacy measures currently employed for low back pain and the aspects of health literacy they include; (2) The low back pain health outcomes included in such work; (3) The extent to which these health outcomes reflect the Core Outcome Set for Clinical Trials in Non-Specific Low Back Pain.The scoping review yielded ten relevant studies. Among the nine different measures of HL used, all involved the study of functional HL. The 50 measures of LBP health outcomes could be grouped into five thematic categories, namely: Pain, Disability, Behaviour, Knowledge and Beliefs, and Resource Utilisation. However, most of these health outcomes did not seek to directly satisfy the COS for Clinical Trials in Non-Specific LBP.The studies included in this scoping review adopted a wide variety of measures to document HL (nine measures used) and health outcomes (50 measures used). This hampered the comparison of results across studies and the development of a comprehensive evidence-base despite the development of the COS .Despite expanding the search and using a more open search criteria, this study only included seven studies in addition to those in the systematic review by Edward et al. . The deaAlthough no restrictions were placed on the year of publication, all relevant studies were published in 2010 or later, suggesting that interest in the association between HL and LBP is relatively recent. This may reflect HL being a relatively new concept within healthcare , and theDespite no language or country restrictions being placed on the search, all studies were conducted in high income countries—as defined by the 2021 World Bank classification of economies . This maFew studies incorporated the four outcome domains of the COS ; only 14 of the 50 health outcome measures used did so. Moreover, these 14 measures were limited to the two core outcome domains of pain intensity and physical functioning , which may be biased by one’s physical and psychological states, along with one’s memory, willingness, and ability to answer the questions. This may influence one’s ability to give accurate self-assessments of health status . This liMany studies also had limitations in terms of the HL measure used. Communicative and critical HL measures were under-investigated. HeLMS, and the questionnaires used by Camerini and Schulz , MacLeodStudy population characteristics were also a source of limitation in the studied literature. Briggs et al. , 41 faceResponder bias through self-selection was another common limitation in terms of study population design. This is important in the context of HL studies, as low HL patients with lesser ability to communicate well with their healthcare provider may have a tendency to decline study involvement . This liThe strengths of this review include the wide search strategy, involving 13 bibliographic databases with no search limiters or filters. By including studies on all forms of LBP health outcomes associated with HL, this review was able to build on the work of Edward et al. . This reNevertheless, the search strategy was not without its flaws. The use of MeSH terms was done with the aim of improving reproducibility of results, especially with searches repeated periodically in this study. However, this ran the risk of missing out on recent articles not yet indexed. Furthermore, a more broadly defined strategy using additional synonyms for MeSH terms could have broaden the search even more. Grey literature was also excluded, which given the paucity of evidence in the field, could well have enriched this study’s findings .The studies by Devraj, Herndon and Griffin , MacLeodThe heterogeneity of measures employed, as well as the paucity of relevant studies, made it difficult to compare findings across studies and provide firm conclusions on the association between HL and each LBP health outcome. Thus, we were unable to draw strong evidence-based conclusions on this. We also note that classifying measurement tools into functional, communicative and critical HL as proposed by Nutbeam  is an imThe protocol was not registered a priori, leading to potential bias. However, as stated, no changes to the protocol were necessitated during the review process and data extraction remained per protocol. Piloting of the data extraction form was also not included. However, these are not requirements of a scoping review and were deemed unnecessary to fulfil the study objectives.In terms of stakeholder involvement, while experts (clinicians and researchers) in the field of HL and LBP are members of the research team, patients were not consulted. There is growing evidence of the value of patient and public involvement at all stages of the research process , and theThe ten relevant studies included in this review yielded a total of nine different measures of HL and 50 measures of LBP health outcomes. Most health outcomes evaluated by the included studies did not seek to directly satisfy the Core Outcome Set for Clinical Trials in Non-Specific LBP. The wide variety of measures used hampers efforts to form conclusive relationships between HL and LBP outcomes, and precludes the use of a meta-analysis approach. To allow for comparison across findings and the development of a rigorous evidence base, future work should seek to address the Core Outcome Set for Clinical Trials in Non-Specific LBP. Furthermore, research thus far has focused on a narrow range of populations and there is an urgent need to broaden the evidence-base to include those where LBP morbidity is high, but data is lacking. As noted above, this is especially so in LMICs. Such work demands the incorporation of comprehensive measures of health literacy that have both generic and culturally sensitive components.


The formation of biogenic amines in food and beverages is mainly due to the presence of proteins and/or free amino acids that represent the substrates for microbial or natural enzymes with decarboxylation or amination activity. Fermentation occurring in many alcoholic beverages, such as wine, beer, cider, liqueurs, as well as coffee and tea, is one of the main processes affecting their production. Some biogenic amines can also be naturally present in some fruit juices or fruit-based drinks. The dietary intake of such compounds should consider all their potential sources by both foods and drinks, taking in account the health impact on some consumers that represent categories at risk for a deficient metabolic activity or assuming inhibiting drugs. The most important tool to avoid their adverse effects is based on prevention through the selection of lactic acid bacteria with low decarboxylating activity or good manufacturing practices hurdling the favoring conditions on biogenic amines’ production. The biogenic amines (BAs) are nitrogenous compounds deriving from enzymatic reactions such as decarboxylation, transamination, reductive amination, and degradation of the corresponding precursor amino acids. Based on their chemical structure, they are distinguished as aliphatic , aromatic (β-phenylethylamine and tyramine), or heterocyclic (tryptamine and histamine). They are also classified into monoamine (tyramine and β-phenylethylamine), diamine , and polyamine  according to their number of amine groups . In TablPseudomonas, Clostridium, Bacillus, and Photobacterium, as well as to the Enterobacteriaceae family , are reported as BA sources. In fermented products, such as cheese, sausages, and fermented vegetables, but also beverages, lactic acid bacteria (LAB) produce BAs to survive on an acidic milieu during fermentation. The genera Enterococcus, Lactococcus, Lactobacillus, Pediococcus, Carnobacterium, and Leuconostoc can decarboxylate amino acids in foods and drinks [The availability of free amino acids, the presence of decarboxylase-positive microorganisms, and further conditions favoring the microbial load development, such as an improper temperature of storage and ripening or fermentation processes, can increase their production. Microorganisms belonging to specific genera, such as d drinks .Some conditions occurring during the fermentation process of foods and drinks can significantly affect and/or increase the synthesis of BAs in the final product. It has been demonstrated that the decarboxylation of amino acids is more active in acid environments where pH ranges from 4.0 to 5.5 and when the temperature is between 20 °C and 37 °C [2) in wines can also reduce both microbial and enzymatic activities [An opposite situation is observed with temperature, i.e., while lower pH correlates with higher BA content, lower temperatures reduce both bacterial growth and decarboxylase activity, and BA production significantly decreases below 5 °C . Additiotivities .The main source of exogenous BAs is the diet through the uptake of foods or beverages containing high concentrations of these compounds. The BAs profile in beverages varies according to the raw material, endogenous enzymes, and microorganisms naturally present or added during the production process. In this review, the main BAs occurring in fermented and non-fermented beverages, their concentrations reported in literature, and the prevention measures able to reduce their presence will be described.The alcoholic beverages are characterized by a fermentation process with production of ethanol and are generally classified as wines, beers, and spirits. The latter contain the highest alcohol content and derive from the distillation of fermented products or maceration of raw material in ethyl alcohol. As ethanol can inhibit the activity of monoamine oxidase (MAO) and diamine oxidase (DAO), a high BA content in such beverages can represent a health hazard delaying the decomposition of these compounds in the human body. Another metabolite of wine named acetaldehyde as well as the anti-depressive drugs based on MAO inhibition, can cause the same interferences in BA metabolism .Saccharomyces cerevisiae) and the malolactic fermentation of LAB are the main processes favoring BA formation [The formation of BAs in wine is associated with the amino acid content, the variety and degree of ripeness of grapes, and the vinification techniques. They can be naturally present in the raw material or formed during the different phases of production  and/or sormation .2), and other minor metabolites. The dominant S. cerevisiae strains, but also other species known as wild yeasts, i.e., Saccharomyces bayanus, Brettanomyces bruxellensis, and Kloeckera apiculata, have been shown to produce Bas, such as histamine, tyramine, spermidine, ethanolamine, agmatine, phenylethylamine, and cadaverine. The non-Saccharomyces yeasts rapidly decrease due to the strong selective pressure exerted by S. cerevisiae [S. cerevisiae to ensure a reproducible, predictable, and controlled fermentation. Wines produced under this practice show low variability, complexity and typicity, and similar sensory properties. By contrast, the spontaneous fermentation may cause some problems to predict their evolution but wines have greater complexity and present higher differentiating characteristics. Moreover, the variety of yeast species within the winemaking environment is associated with the cellar resident population on wall surfaces, equipment, and oak barrels that play an important role in the wine aroma complexity. Thus, the cellar cleaning and hygienic practices can influence the winery microbiota affecting their diversity, composition, and evolution [The alcoholic fermentation (AF) is mainly carried out by yeasts converting sugars into ethanol, carbon dioxide  is the second important step in winemaking that is carried out by LAB species. In this process, the malic acid is converted in lactic acid and COconostoc . They caroducers . Finallyroducers .2 concentration, clarification treatments, microorganism strains, and the aging period [2, or the use of malolactic bacteria strains that have been screened for the absence of BA-producing genes [Some other enological practices can also affect the final content of BAs in wine, such as SOg period ,11. A pHg period . Some wing genes .The most BAs in red and white wines reported in literature are shown in Pediococcus damnosus, Lactobacillus frigidus, Lactobacillus brevissimilis, and L. brevis) have been described as amine-producing microorganisms in beer. Finally, BAs can also be formed during storage [Beer is a carbonated alcoholic beverage consumed worldwide. The most common sources of BAs in beer are the raw material  and the contaminating microflora (mainly LAB). Histamine, cadaverine, putrescine, 2-phenylthylamine, and tyramine are the most detected BAs produced by decarboxylase positive microorganisms, while spermidine, 2-phenylethylamine, and spermine can be naturally found in malt, wort, and hops. Putrescine can show both origins. Moreover, the prolonged storage of the product can favor BA formation in beer . With re storage . Lorenco storage  reportedOenococcus, Lactobacillus, Pediococcus, and Enterococcus, are often able to produce BAs including tyramine, putrescine, and cadaverine [Other types of wine are the so-called fruit wines which are fermented alcoholic beverages made from a variety of ingredients other than grapes and/or additional flavors from fruits, flowers, and herbs. Płotka-Wasylka et al.  analyzeddaverine . Rice widaverine . It is odaverine .Apis mellifera honey as well as spermine and tyramine in those produced with honey from Melipona quadrifasciata anthidioides by using different finings and storage systems in oak barrel. In greater detail, when fruit peels  were used as finings, a BA increase was observed only in the first situation, i.e., the above-mentioned BAs were found in banana/Apis mead probably derived from the banana peel, while only spermidine was detected in the control samples. By contrast, the mead from passionfruit/Melipona did not show differences for BA content with the control group and therefore the use of this fining did not generate any new compounds. Liqueurs are spirit beverages containing alcohol and sugar at different percentages and obtained through different methods, i.e., through the infusion of herbs or roots in hot water and alcohol, cold maceration of fruits or flowers with distilled alcohol, or distillation of alcohol and flavoring agents. Various industrial and homemade liqueurs made from fruits, herbs, coffee, honey, and milk were analyzed by Cunha et al. (2017) [Mead is an alcoholic beverage produced by the fermentation of honey and water. Silva et al. (2020)  detected. (2017)  for BA pVinegar fermentation usually occurs after alcohol fermentation through the oxidation of ethanol by AAB. The traditional Bokbunja vinegar is produced from black raspberry through a two-step fermentation of alcohol and vinegar, and it is well known in Korea for its functional ingredients such as flavonoids, tannins, and phenolic compounds. During its production, many BAs can derive from both fermentation processes. Song et al.  isolatedCoffee is one of the most popular drinks worldwide, ranking second only after crude oil. It can be produced by three different extraction techniques, i.e., decoction, infusion, and pressure. The most represented BAs in coffee beans and coffee beverages are putrescine, spermidine, spermine, and serotonin, while cadaverine and tyramine can also be present but at smaller concentrations . The proCamellia sinensis) leaves undergo heating or steam treatment and fast drying, while during black tea production, they are subjected to weathering, various rolling, and crushing and/or tearing processes, followed by enzymatic maturation, and then final drying. By such a way, some enzymatic and oxidative reactions are responsible for BA formation, achieving levels of 20 and 14 mg/L for histamine and cadaverine, respectively [Another stimulating beverage that is very much consumed worldwide is tea. According to its production, it can be classified as unfermented (green tea), semi-fermented (oolong tea), fully fermented (black tea), or post-fermented (pu-er tea). To obtain green tea, the tea . The presence of BAs in such drinks can be linked to the food matrix of origin  or to inadequate hygienic conditions and/or microbial contamination . The vegAccording to the production process, fruit juices can be made from fresh fruits without flavorings, colors, or other added ingredients, while fruit nectars are obtained from the addition of water, with or without added sugars, to fruit juices . As polyBiogenic amines can be both essential and harmful to human health. According to their physiological effects, they are classified in vasoactive and psychoactive amines. The former, i.e., tyramine, tryptamine, and histamine, are involved in blood pressure control, whereas the psychoactive amines  act on the synaptic transmission. The consumption of foods and beverages with high BA concentrations can cause some adverse effects such as migraine, headache, bowel disorders, and allergic reactions. The latter are known as scombroid poisoning and cheese reaction due to histamine and tyramine, respectively. Putrescine and cadaverine cause hypotension and bradycardia, besides potentiating the toxicity of other amines. Moreover, high amounts of putrescine, spermidine, and spermine can accelerate the development of cancers, as they have been found in tissues with high growing rates . Such po2 or lysozyme to control undesirable LAB in the musts, and the use of clarification products such as bentonite and fining agents. During clarification, some nutrients and suspended particles that can favor the growth of bacteria are removed. Then, microbiological analyses able to check yeasts and LAB population in must and wine should also be performed. Finally, good hygiene practices, temperature control, and proper storage are the main prevention actions for all beverages, other than wine, in which BAs can be formed. As histamine is generally not found in the raw material intended for beer production, it can be considered a good indicator of brewing hygiene and the bacterial contamination during all the production phases should be regularly controlled to ensure that the final product meets all quality and safety standards. Moreover, the under-pasteurization of beer can also increase the BA content [The European Food Safety Authority proposed potential Acute Reference Doses for the following BAs, i.e., 50 mg for histamine in healthy individuals but below the detectable limits for those with histamine intolerance; 600 mg for healthy individuals; 50 mg for those taking third-generation MAO inhibitors drugs; or 6 mg for those taking classical MAO-inhibitor drugs. According to the Food and Drug Administration, the tolerable level of histamine corresponds to 50 mg/Kg . Neverth content . In non-