Objective: Cognitive impairment in schizophrenia forms the key cause of the disease's disability, leading to serious functional, and socioeconomic implications. Dopaminergic-cholinergic balance is considered essential to cognitive performance in schizophrenia and patients are often treated with many drugs with anticholinergic properties. This study aims to examine the cognitive impact of anticholinergic burden in patients with schizophrenia.Methods: A systematic literature review was performed on English-language studies published on PubMed, Embase, and Web of Science, from inception to June 2021, to identify research studies that examined the effect of anticholinergic load on cognition in clinically stable patients with schizophrenia. No restrictions on study design, age of participants, or geographical distribution were applied. Two researchers performed independently the screening and shortlisting of the eligible articles. A narrative synthesis of the main characteristics and findings of studies included was reported.Results: In total, 17 articles of varying methodological design met the inclusion criteria. Three of them found statistically significant improvement in cognition after anticholinergic tapering without adverse effects. Thirteen studies found a statistically significant association between high anticholinergic burden and cognitive impairment , apart from a study, related to the specific characteristics of clozapine.Conclusions: Medication with increased anticholinergic load has been found in most of the studies to negatively affect neurocognitive performance of patients with schizophrenia. However, the clinical and methodological heterogeneity of studies included limit our interpretation and conclusions. Cholinergic neurotransmission plays a crucial role in both psychotic symptoms and cognitive disorders as well as treatment of schizophrenia . SpecifiIn addition to findings that correlate the pathogenetic mechanism of the disease with abnormal cholinergic neurotransmission, patients with schizophrenia are often treated in clinical practice with anticholinergic drugs to control the extrapyramidal side effects that cause most antipsychotics, especially first generation –19. FurtGiven that impaired cognitive performance is a strong determinant of the outcome of the disease as well one of the major causes of the disease's disability affecting the daily social, occupational functionality, autonomy, and independence , 24, theFor the conduction of this systematic review, we followed the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines (PRISMA) .A systematic literature search in PubMed, Embase, and Web of Science was conducted by two independent authors (RG and KG) to identify articles that examined the effect of anticholinergic load on cognitive domains in patients with schizophrenia. The following terms were used: (anticholinergic OR antimuscarinic OR parasympatholytic) AND (cognitive OR brain OR memory OR dementia OR confusion) AND schizophrenia. All databases were searched from their inception through June 2021, restricted to English language publications. First, the title and abstract of each article was examined, and then the full texts of potentially eligible articles to be included in the systematic review were evaluated. A reference list of relevant studies was screened to identify additional studies.We used the Population, Intervention, Comparison, Outcomes and Study design (PICOS) approach  for the Assessment of exposure's degree to drugs with anticholinergic properties is a combination of the daily dose, the affinity to muscarinic receptors, the permeability to the blood barrier, and drugs' general pharmacokinetic and pharmacodynamic characteristics. Therefore, many different scales and methods of classifying cumulative anticholinergic burden are available. Eligible intervention criteria were (a) the tapering of anticholinergic therapy or (b) the cumulative anticholinergic load ranked either by anticholinergic classification scales or calculated based on serum anticholinergic activity (SAA).in vitro pharmacological properties. Medicines with anticholinergic properties are ranked from 0 (none) to 3 (high) (in vitro binding affinity with muscarinic receptors. Drugs are graded from 0 (no anticholinergicity) to 3 (high anticholinergicity with significant adverse cognitive effects) : A classification system for rating anticholinergic potency of medicines based on the clinical experience, expert opinion, and 3 (high) , 28. Anteffects) . Antichoeffects) . SAA: Seeffects) .Since cognitive impairment in schizophrenia is observed in a variety of cognitive domains  different diagnostic tools can be used for cognitive performance evaluation. The MATRICS Consensus Cognitive Battery (MCCB), although an FDA-approved neurocognition test battery, is usually only used to extract an overall score, whereas the assessment of complex domains such as executive function requires additional diagnostic tools  . ComprehTwo authors (RG & KG) extracted data independently and any discrepancy was resolved by discussion with the third author (DL). A standardized data extraction form was developed for our systematic review and it was used to minimize data entry errors and improve validity and reliability. The extracted data included first author, publication year, country of origin, design of the study, sample size, characteristics of the study population, study groups, considered confounding variables, and summary of findings.The methodological quality of cohort and case-control studies was performed by the Newcastle-Ottawa Scale (NOS) . The NOSFrom the initial electronic search, we found a total of 734 articles, 679 of which were excluded based on title and abstract review. The full text of the remaining 55 was checked for eligibility and 38 of them were excluded. Overall, a total of 17 articles were included in this systematic review . No furtThe publication years were from 1982 to 2021. The countries covered are the United States, China, South Korea, Germany, Canada, Japan, and Israel. The number of participants included in the studies ranged from 15 to 1,120. Most of the studies included, recruited clinically stable adults in inpatient setting, with no history of pre-existing organic impairment, unstable medical or neurological conditions, or prohibited drug abuse. As regards to the study design, the majority were cross-sectional studies –47. Addin = 27, n = 34, and n = 20), after successful discontinuation of anticholinergic treatment by most participants, no adverse effects to psychopathology or extrapyramidal symptoms were found, while a significant improvement in the composite/overall scores of the neurocognitive batteries used  during the follow-up weeks was observed. Anticholinergic tapering showed improvement over baseline in cognitive tasks that mainly assess the domains of processing speed, attention, ideational Praxis, and Orientation.The characteristics of studies included that examine the association of anticholinergic tapering with neurocognitive performance are presented in p = 0.029, p = 0.001, and p = 0.002, respectively), with average corresponding differences −0.287 (95% CI: −0.023 to −0.552), −0.416 (95% CI: −0.146 to −0.686), and −0.517 (95% CI: −0.163 to −0.871), whereas in the Ogino et al.  = 6.06, p = 0.02], 4 weeks after discontinuation. In both studies improvement was observed in tasks that incorporate the domains of attention and processing speed. Particularly in the Desmarais et al. (p = 0.043) with an average improvement of −0.279 and 95% CI from −0.006 to −0.552. As regards the Ogino et al.  = 4.75, p = 0.04] and regarding the analysis among the participants of the biperiden discontinuation group, significant time changes in verbal fluency  as well as in symbol coding task  were observed. In the overall interaction (time × groups), also a statistical significance appeared in the symbol coding test . Drimer et al. , orientation , and overall score  10 days after anticholinergic discontinuation.In the Desmarais et al.  patientso et al.  in the os et al.  significo et al. , in the r et al.  used a dOut of the six studies that evaluated the estimated overall cognitive performance with the combination of representative cognitive tests from multiple domains, five provided evidence that increased medication anticholinergic burden was significantly associated with decreased global cognition or composite scores comprised of several different cognitive tests.r = −0.512, p < 0.001). The correlation was also confirmed through regression analysis . Although the explanatory power decreased after adjusted analysis, anticholinergic load continued to have a statistically negative impact on the MCCB composite score (p = 0.013). The recent study of Joshi et al. with a large sample size (p = 0.004) evaluated with the BACS, by approximately a mean of 0.58 lower SD, than patients with an ADS score <4. In addition, an also large study in terms of the number of participants (n = 705) found that the anticholinergic burden had a significant negative correlation with cognitive performance in global cognition however the effect size was small for anticholinergic burden scale (ABC), (Cohen f2 = 0.008) and for anticholinergic drug scale (ADS), (Cohen f2 = 0.017), and therefore the clinical significance of the result is not apparent  represented 20% of the variation in global cognition change, regardless of age, IQ, or severity of symptoms. On the other hand, the study of Tracy et al. (p < 0.001) treated with antipsychotic clozapine compared to those treated with antipsychotic risperidone (lower SAA).Specifically, in the study of Kim et al. , in whicple size  was alsople size  detectedapparent . The difapparent  also proIn total, six studies with different methodological design and a variety of medication anticholinergicity measures reported significant association between increased anticholinergic burden and decline in several types of memory  as well as the domains of learning and language/verbal skills.in vitro indicators reported a significant association of high anticholinergic load with worse declarative memory performance. Specifically, the study found that an increase in anticholinergic load from 0.5 to 4 mg/day equivalent pharmacological benztropine can lead to a reduction of up to 1.7 SD of the California verbal learning test performance. In addition, Sweeney et al. , with which after regression analysis, SAA showed a common variation of 7%, after controlling for the impact of age, IQ, and severity of symptoms. In addition, the cognitive effects of anticholinergic load, evaluated according to a different anticholinergic scale , in older patients with schizophrenia (≥50 years), bear a similarity to cognitive dysfunction that occur in the early stages of Alzheimer's disease. High anticholinergic load was correlated with short-term memory decline (p = 0.004) evaluated with Alzheimer's Dementia Battery and the Repeatable Battery for the Assessment of Neuropsychological Status. It is also worth noting the emergence of a correlation tendency between high anticholinergic load and poorer learning performance , which refers to the mental process of retrieval of the information/events that were previously encoded and stored in the brain, in total four studies found a significant negative association with high SAA.r = 0.51, p < 0.01) demonstrating a negative effect of the high anticholinergic load on recent or working memory. On the other hand, a cross sectional study (n = 17) of chronic patients, similar age range but serum anticholinergicity due more to neuroleptic treatment than to antiparkinsonian drugs, was only partly in line with Tune's et al. . As regards the recognition memory, which was evaluated with the Mattis-Kovner inventory test, no association with the anticholinergic load  was found. Recall performance also declined  in the group with the high serum anticholinergic load in the study of Tracy et al. (r = 0.40). In addition, the Hitri et al. . Furthermore, high anticholinergic load assessed with ACB was also correlated with worse performance in spatial working memory (p = 0.04) in the study of Tsoutsoulas et al. , in which after statistical analysis, the task showed common variance with serum anticholinergicity of 19% (r = −0.45). Similarly, in the Ang et al.  but is likely to be symptomatic as no significant results were observed under the increasing CDD load neither in the whole sample nor in the subgroups.In addition, in the Tracy et al.  patientsg et al.  executivFurthermore, the domains of processing speed , divided attention, and visual-spatial ability have been reported to be negatively associated with high medication anticholinergic burden.b = 0.292, p < 0.05), as opposed to treatment with antipsychotics without anticholinergic properties. With the application of locally weighted scatterplot smoothing tool to verify the relationship of cognitive function with the increase in anticholinergic daily dose, although the tendency for reduced speed of information processing was confirmed by increasing CDD, a paradox was observed, since the group of the highest anticholinergic dose (BZT-Eq > 20) exhibited a higher rate of information processing than the group of 0 < BZT-Eq < 15. Nevertheless, this could be attributed either to insufficient data on anticholinergic equivalent doses for certain drugs or to the specific pharmacological properties of clozapine taken by all patients as the major antipsychotic drug, which could compensate for the adverse anticholinergic effects. Poorer processing speed/vigilance performance was also associated with the high anticholinergic burden in the study of Ang et al.  in Sweeney et al.  with clig et al. , howeverp = 0.02) evaluated with Alzheimer's Dementia Battery and the Repeatable Battery for the Assessment of Neuropsychological Status in Tsoutsoulas et al.  that will examine the longitudinal exposure in anticholinergic agents, with a more representative sample and longer monitoring duration to eliminate as many systemic errors and biases as possible and to draw more universal conclusions. More research is also certainly needed from a neurological point of view to explain and clarify the involvement of cholinergic neurotransmission and the general neurochemical mechanisms in the cognitive impairment of people with schizophrenia, since there are several hypotheses about various pathophysiological mechanisms that involve different neurochemical pathways and neurotransmitter systems including the role of muscarinic and nicotinic acetylcholine receptors , 76–78. As far as clinical practice is concerned, it is important for clinicians and other health professionals to assess the cumulative effects of anticholinergic drugs on cognition. Specifically, neurocognitive deficits can even explain about 20–60% of the variation in functional performance observed among patients with schizophrenia , 86–88. One of the main limitations of the studies included is the small sample size of the participants. Furthermore, amongst prospective cohort studies , 49, theIn terms of the present systematic review limitations, no meta-analysis could be performed due to the heterogeneity in the populations, methodology, and interventions of the included studies. Therefore, this affects the quality of the evidence presented because it is unclear whether the positive studies were favorable or whether there was a performance bias. Language bias is also possible as we could have missed non-English studies. Moreover, according to the hierarchy of evidence, most of the studies included on their methodological design are cross-sectional or retrospective studies with a limited number of participants. Hence, there is no time sequence between the exposure and the disease-outcome studied and cannot safely prove a causal relationship.in vivo assessment of the central effect of anticholinergic medication. Furthermore, as a tool it cannot be used to draw conclusions about which medicine in particular may need to be discontinued to reduce the anticholinergic load, while the cost and availability of the method lead to the use of anticholinergic drug scales, which also have certain limitations (It is also worth mentioning that the studies included were based on tools which vary in terms of the classification of the anticholinergic burden of active substances, and which also have several limitations. Although the method of measuring SAA is the gold standard method for assessing anticholinergicity, this may reflect only a transitional cholinergic condition outside the brain, which confers an intuitive clinical capability but lacks a direct itations , 41. In itations , 92.The present systematic review shows that medication with increased anticholinergic load is possible to affect the cognitive functions of people with schizophrenia. However, based on different research methodologies and the clinical heterogeneity among various studies, it is not reasonable to make a definitive conclusion. Well-designed large prospective studies and randomized clinical trials are required to examine the effect of anticholinergic drug treatment on cognition in patients with schizophrenia. Based on these findings, clinicians are required to reconsider the need for anticholinergic treatment, with caution in prescribing medicines with known anticholinergic activity, before recommending any treatment for cognitive rehabilitation in schizophrenia.The original contributions presented in the study are included in the article/supplementary material, further inquiries can be directed to the corresponding author/s.RG carried out literature searches, appraised the articles, summarized the results, prepared the tables and figures, wrote the manuscript, and interpreted the results. DL interpreted the results. KG supervised the process, carried out literature searches, appraised the articles, summarized the results, wrote the manuscript, and interpreted the results. All authors contributed to the article and approved the submitted version.The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Mycoplasma pneumoniae pneumonia (MPP).To investigate the correlation between serum immunoglobulin E (IgE) levels and the complications in children with A retrospective study of MPP patients hospitalized from May 2019 to July 2021 was performed. We analyzed the clinical manifestations, complications, laboratory findings, and treatments.p = 0.041. Patients with complications of necrotizing pneumonitis, pneumothorax, skin rash, or bronchiolitis obliterans had higher IgE levels. There was no statistically significant difference in IgE levels between pulmonary complications and extrapulmonary complications. The older the age, the greater the probability of complications (p = 0.001). The group with complications was more likely to have chest pain (p = 0.000), while the group without complications was more likely to have wheezing (p = 0.017). The use of bronchoscopy and glucocorticoids was higher in the complication group than in the non-complication group (p = 0.000).A total of 275 patients who met the inclusion criteria were enrolled in the study. We divided patients into two groups based on whether there were complications. Complications occurred in 147 patients, of which pulmonary complications were more common than extrapulmonary complications. The IgE level in the complication group was higher than that in the non-complication group with MP infection.MPP patients with higher IgE levels had more severe clinical symptoms and complications. We speculated that IgE might be a biomarker for complications after  Mycoplasma pneumoniae (MP) is a common pathogen of community-acquired pneumonia in children  is mostly self-limited, but it can produce a variety of pulmonary and extrapulmonary complications and can even be life-threatening in some individuals  increased in children with MP infection. Some studies also found that MPP patients had elevated total serum IgE levels  met the diagnostic criteria: with clinical manifestations , abnormal lung auscultation, radiologic findings (presence of a new infiltrate on chest radiography or consolidation not attributable to some other etiologies), and ry tests ; and 2) ry tests .MP antibody . It was defined as MP positive when the MP antibody titer was ≥1:160. Nasopharyngeal aspirates were collected on the day of admission, and MP-DNA was detected by PCR . Simultaneous detection of other pathogens was performed through the indirect immunofluorescence assay of respiratory viruses  using a D3 Ultra DFA Respiratory Virus Screening & ID Kit  and blood cultures for bacteria (BD FX200 blood culture system).On the day of admission, 3 ml of venous blood was collected from patients, and the passive agglutination assay was used to detect By consulting the electronic medical records of all patients, the demographic, clinical, and laboratory data were collected retrospectively. Clinical signs and symptoms of patients, including fever, cough, wheezing, chest pain, and complications, were obtained. All patients underwent chest X-ray or chest CT scan to confirm focal or segmental infiltration, with or without pleural effusion, atelectasis, pneumothorax, pulmonary embolism, and pulmonary necrosis.Serum IgE levels were obtained using the automatic biochemical immunoassay analyzer produced by Roche. The test kit was also provided by Roche. The reaction was carried out according to the manufacturer’s instructions.This study was approved by the Ethics and Research Council of Women and Children’s Hospital of Ganzhou (201905-A01) on May 15, 2019. The data from patients were collected anonymously.p-Values <0.05 were considered to be significant. The figures and graphs were generated by GraphPad Prism 5.The statistical analyses were carried out using SPSS 20.0 software package. Continuous variables were reported as the mean ± SD and were compared using Student’s t-test or the non-parametric Mann–Whitney U-test. The categorical variables were shown as number (%) and were compared using the chi-square test. From May 2019 to July 2021, 275 patients admitted to our hospital for MPP were enrolled in the study. The demographic and clinical characteristics are shown in p = 0.292, as shown in The pulmonary and extrapulmonary complications are listed in p = 0.001). No statistically significant difference in the appearance of fever and cough symptoms was observed between the two groups. Higher incidence of chest pain (p = 0.000) and lower incidence of wheezing (p = 0.017) were found in the complication group than in the non-complication group. Significant differences in fever duration, hospitalization duration, and incidences of SMPP between the two groups (p < 0.001) were also found. The probability of SMPP in patients with complications was higher than that in patients without complications (p = 0.000). As shown in p = 0.041). The risk factors for complications by multifactor binary logistic regression analysis were analyzed. After age factors were adjusted, a significant difference in the IgE level was noted between the complication and non-complication groups (p = 0.047), as shown in p < 0.01). However, no significant difference was observed in white blood cell count, neutrophil percentage, hemoglobin, platelets, erythrocyte sedimentation rate, and the levels of immunoglobulin A, immunoglobulin G (IgG), immunoglobulin M, alanine transaminase (ALT), aspartate transaminase (AST), and creatine kinase isoenzymes (CK-MB). Patients in the complication group were more likely to receive oxygen therapy, glucocorticoids, and bronchoscopy as compared with those in the non-complication group (p < 0.01). Meanwhile, eight patients required ICU treatment, and one required mechanical ventilation. Except for one child who died, the rest of the children recovered and were discharged from the hospital.Patients were divided into two groups according to the presence or absence of complications. The demographics, clinical characteristics, laboratory findings, and treatments of children in the complication and non-complication groups are shown in There were some patients with abnormally increased IgE  in our study. The information of these 14 patients is listed in MP infection has increased in recent decades. Few studies reported the incidence of various complications caused by MP. Most studies were in the form of case reports or reviews. The incidence of pulmonary complications in this study was 34.5%, which was slightly higher than that reported in the literature. Xu reported that the incidence of pulmonary complications in their study was 20.65% (MP infection could reach 25%–50% (MP. We found that 7.64% (21 in 275) patients had hypokalemia. Han reported that 15.7% (77 in 489) patients had hypokalemia, which was associated with SMPP, fever duration, and hospitalization duration . Written informed consent to participate in this study was provided by the participants’ legal guardian/next of kin. Written informed consent was obtained from the individuals, and minors’ legal guardian/next of kin, for the publication of any potentially identifiable images or data included in this article.LZ contributed to the concept and assisted in the critical writing. YL contributed to design of the study. ZX and XP helped in analyzing and interpreting the data. XG and LY contributed to the collection of clinical information. All authors contributed to the article and approved the submitted version.The project was supported by grants from Bethune Foundation (SCE041BS).The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Phalacrocorax carbo). The samples were collected from Central Tisza-Jászság Nature Conservation Area in Hungary. The tissue samples were analysed by inductively coupled plasma optical emission spectroscopy (ICP-OES). The aim of the study was to examine the impacts of heavy metal pollution on the water birds, determine the concentrations of the abovementioned metals in the different muscle tissues of these wetland birds, and provide the basic materials for monitoring the environmental pollution. Among the investigated elements/metals, the detected concentrations of As, Ba, Cd, Co, Cr, Mo and Ni were below the detection limit. Higher concentration of Cu, Hg, Mn and Pb was measured in the pectoral muscle compared to the thigh muscle, but only in the case of Cu and Mn were found significant differences between the tissues. In the case of the Zn concentration, the higher value was detected in the thigh muscle. There were no statistical differences between males and females in either metal concentrations.Concentrations of 12 metals  were examined in the pectoral and thigh muscle of great cormorants ( Due to the rapid development of toxicology, more and more researches and studies have been published that have reported the pronounced toxicity of various compounds of metals. Based on the results, the use of many metal compounds as pesticides has been banned by the licensing authorities. However, due to anthropogenic activities  and urban pollution , the entire biosphere has become contaminated in different levels EFSA . At the Anas spp.), cormorants , gulls (Larus spp.), white storks (Ciconia ciconia), common buzzard (Buteo buteo), feral pigeon (Columba livia domestica) and eagle owl (Bubo bubo), which are located on higher trophic levels in the ecosystem, are widely used as bioindicators for monitoring metal contamination in the environment . Metals are especially excreted via urine and faeces (through biliary clearance), but other routes may be probable  this proportion can be as high as 80%  at area of the Central Tisza-Jászság Nature Conservation Area of the Hortobágy National Park Directorate, and thus the possible metal pollution in the area, and to obtain information, similarity or difference on metal accumulation in the two types of muscle tissues.In our study, we aimed to investigate the heavy metal pollution  of the pectoral and thigh muscle of great cormorants , under the supervision of the Nature Conservation Service, twenty cormorants were shot as a population management activity to reduce the overall cormorant numbers in the region. These birds were collected after being shot with official permission , based on Decree No. 13/2001 of the Minister of Environment  were taken during the necropsy as well. Twenty grams of both muscle tissue samples was placed into an individually labelled plastic bag. All samples were placed in a cooler bag and transported to the laboratory where they were stored at −20°C until assayed.3/min; cooling water flow rate, 1 dm3/min; sheath gas flow rate, 0.9 dm3/min; sample feeding flow rate, 0.9 cm3/min; and observation height, 15 mm.The heavy metal concentration of the samples was determined in the analytical laboratory of the Department of Animal Hygiene, University of Veterinary Medicine, using a Perkin Elmer Optima 3300 DV inductively coupled plasma optical emission spectrometry (ICP-OES) as described by Grúz et al. . During 3) and hydrogen peroxide (H2O2) and a microwave digester were used to extract the samples.Nitric acid . The measurements were performed with argon gas of 4.6 purity (Messer Hungarogáz Kft).For sample digestion, 0.5 g from each sample was weighted into a CEM MARS6 MARSXPreSS Teflon vessel. Then they were decomposed by 5 ml nitric acid (69m/m%) and 5 ml hydrogen peroxide (30m/m%) in a microwave digestion system . The sample was filled up to 25 ml and then diluted twice.The following metals were determined from the samples: arsenic (As), barium (Ba), mercury (Hg), cadmium (Cd), cobalt (Co), chromium (Cr), manganese (Mn), molybdenum (Mo), nickel (Ni), copper (Cu), lead (Pb), zinc (Zn). During the measurement, the limit of detection (LOD) of each metal was as follows: 0.05 μg/g for Cd, Co, Cr, Cu, Mn, Zn; 0.2 μg/g for Ni, Pb; 0.5 μg/g for As, Ba, Hg, Mo.Statistical analyses were performed using SPSS (version 25.0).p values. This regulates that no more than 5% of the pairs found to be significant are misclassified. After the correction, there was no significant difference between the sexes in either case.Given that in many cases we have censored data , sex comparisons were performed using the Mann-Whitney test. In the comparisons, due to the large number, a Benjamini-Hochberg correction was applied to the As there was no significant difference between the sexes in either case, the 10 female and 10 male birds were treated together, as a sample of 20 elements. Due to the censored data, the comparison of tissue samples was performed with a nonparametric Friedman test. This compares the data of an exact tissue sample with all the other tissue samples of the same bird. Pairwise comparisons were then performed using the Wilcoxon test using Bonferroni correction for each body part separately.Sander lucioperca) and Wels catfish (Silurus glanis), and two omnivorous species, Danube streber (Zingel streber), and Black bullhead (Ameiurus melas).In the stomach content, the following fish species were found in the highest quantity: two carnivorous species, Zander  and the mean concentrations were above the LOD, and were higher in the pectoral muscle, than in the thigh muscle (except the Zn concentration) Table .Table 2No gender differences were found for pectoral and thigh muscle. During the examination of the pectoral and thigh muscles, outstanding lead concentration was detected in 1–1 cases. Excluding them from the evaluation, there was no gender difference.Significant difference was obtained for each element, suggesting that each metal accumulates in different concentrations in different type of muscle tissues. Among the investigated metals, the detected concentrations of Cu and Mn were significantly higher in the pectoral muscle than in the thigh muscle Table .Table 3Among the investigated elements/metals, the detected concentrations of As, Ba, Cd, Co, Cr, Mo and Ni were below the LOD. The measured values of Cu, Hg, Mn, Pb and Zn were above the limit of detections; thus, they are discussed.P<0.001). Generally, there is lack of information in the literature about the comparison of Cu in both muscle tissues. However, Carpene et al.  than in its thigh muscle. Since the structure of these muscle types are similar, the size of them can be an explanation of the higher metal content, but more specific further examinations are needed to be carried out in this field.Cu is essential in low concentration for the normal growth, cell metabolism, haemoglobin formation, antioxidant defence system and structure and the function of many proteins . Similar comparison was not found in the literature, likewise in the case of Cu.Manganese is an essential element and one of the least toxic of the essential elements NRC . The accThe main source of lead for waterfowl and other wetland species are the various lead shots, and lead weights used in sport fishing  than in its pectoral muscle. Even if the concentration in thigh muscle was higher than that in the pectoral, opposite like in the case of the other metals, these results were not significantly different.Zinc is an essential element, and it is required for DNA replication, transcription and a cofactor for gene regulatory proteins  of waterfowl and terrestrial bird species of both sexes  were monitored by Grúz et al.  were significantly higher than female ones.Previously, toxic metals  in the feathers of 123 predatory birds  in Hungary , Neotropic cormorant  and, of omnivorous bird, Great kiskadee (Pitangus sulphuratus) were measured in Argentina, without differences in their levels between males and females  at Arctic region . The concentrations of Cu in the muscle and kidney were differed significantly only between the sexes  , Black-headed gull (Larus ridibundus) and Black coot (Fulica atra) in Italy. Zn concentrations were higher in the leg muscle than in the pectoral muscle, but higher levels of Cu and Fe were detected in the breast muscle compared with thigh muscle. However, differences were not stated between genders  in Iran  and Common Teal (Anas crecca) in Iran. Sex difference was noted only in the liver of males.Sinkakarimi et al.  studied The heavy and essential metal burden in the birds’ body is mostly determined by metal accessibility, food quality and environmental pollution presented in their nesting and feeding site.Based on our results, the detected concentrations of these metals accumulated in pectoral and thigh muscle of great cormorants do not indicate that the habitat of the birds is contaminated by them on a level, which would cause toxicosis in the animals, or in humans, since there is a possibility that they consume the same fish species as the birds.So, a routine monitoring of wildlife tissues in various areas is important to collect information about the metal burden of a region .


Human personality influences the way people interact with dogs. This study investigated the associations between the personality of animal shelter volunteers and behavior during on-leash walks with shelter dogs. Video recording and a canine leash tension meter were used to monitor the on-leash walking. Personality was measured in five dimensions  with the NEO Five-Factor Inventory (NEO-FFI). Neurotic volunteers pulled the leash harder and tended to interact with dogs using more body language; dogs being walked by neurotic volunteers in turn displayed more lip-licking and body shaking and were more likely to be rated as well-behaved. Extroverted volunteers were associated with stronger maximal leash tension at both the human and dog ends of the leash, and they praised the dog more, often in a high pitched voice. These volunteers eliciting more tail-wagging and body shaking by the dog. Extroverted volunteers were also more tolerant of different dog behaviors. Volunteers with personalities characterized by “openness to experiences” were less likely to verbally attract the attention of dogs, praise dogs and talk to them in a high-pitched voice; however, dogs walked by these volunteers were more likely to pull on the leash, and engaged in more lip-licking but less sniffing. “Agreeable” volunteers liked to verbally attract the attention of the dogs and more commonly initiated hand gestures and physical contact, causing the dogs to pull less frequently; dogs in these dyads displayed more gazing and lip-licking behaviors. Conscientious volunteers were less likely to pull the leash and tended to have more physical contact with the dogs but did not favor verbal communication and did not use a high pitched voice. Canis familiaris) show high sociability toward humans due to domestication and artificial selection, which enables them to share close relationships with us , of The University of Queensland.The research was conducted at the Royal Society for the Prevention of Cruelty to Animals, Queensland  shelter. Dogs were housed individually in rows of adjacent kennels (1.8 m wide × 1.2 m long × 3 m high) indoors and were able to make visual but not physical contact with each other across the central passage. Every enclosure was furnished with a metal crate, a raised mattress, a water bowl and enrichment . Each dog was walked twice daily, once in the morning from 08:00 to 10:00 h, and once in the afternoon from 14:00 to 16:00 h. The walk started from the kennel and ended with return to the kennel, taking about 15 min to complete. Moreover, dogs had plenty of time interacting with shelter volunteers, staff and visitors. These interactions provided the dogs with abundant social stimuli and enrichment. In this study we only recorded the interaction when dogs were on the designated pathway, with dogs walked uni-directionally, so as not to be confounded by turning around. The time on the designated pathway was ~5 min. The pathway ground had several sections with different coverings to provide tactile and olfactory enrichment for the dogs. The first 40% was covered with gravel, followed by 20% on a concrete section, then 5% on wooden boards, and finally the last 35% was covered with earth. Equipment and infrastructure provided added stimulation and play including two bridges, two dog jumps, some tennis balls and some hanging plastic milk bottles.This study investigated 370 walks from July to early October 2019, involving 111 shelter dogs and 74 volunteers. All participating dogs had to have been resident at the RSPCA, QLD, for at least a week to enable them to become accustomed to the living and walking areas. Dogs' walking behavior was categorized into four levels by RSPCA animal attendants who had been closely working with those dogs. Levels were assigned according to the ease of walking the dogs and was based on their performance during the daily walk. Level 1 dogs walked on a loose leash most of the time. Level 2 dogs pulled on the leash during the walk occasionally and had more undesirable behaviors than level 1 dogs. Level 3 dogs tended to pull on the leash fiercely due to excitement or timidity. Level 3+ dogs had severe behavioral issues, such as overt excitement or fearfulness; however, they were not believed to necessarily pull on the leash harder than level 3 dogs. Dogs with severe behavioral or medical issues that might affect the experiment were excluded from the study because of safety and welfare concerns. All included dogs had undergone an RSPCA behavioral assessment  was commissioned for this project  . Prize draws for $20 RSPCA World for Pets vouchers were offered as incentives. Since there were very few level 1 dogs available during the research period, only volunteers with level 2 and above training levels were recruited. All participants had sufficient English proficiency to follow the research instructions and complete required documents. The research process was explained before the observational study and participants were informed on the research consent form that the overall study aim was to improve the interaction between volunteers and shelter dogs.Dogs classified at the different levels were matched to volunteers of the right experience and training level by RSPCA staff. Dogs were walked on a designated pathway away from distractions at the shelter. Before every walk, the researcher held the leash tension meter vertically downward for 10 s without connecting it to the dog. The signals generated were later used to calibrate the recorded data using MATLAB®. The volunteer connected the leash to the dog's collar and harness in front of its chest via the leash tension meter. A laptop  was carried by the volunteer in a backpack for data collection, and a camera  was mounted on the volunteer's head to record the interaction. Before walking, the volunteer was directed to pull the two ends of the device and hold the pull for 3 s by counting slowly “1, 2, 3.” This procedure was repeated three times, in order to synchronize the tension data with the video. During the walk, the volunteer was instructed not to touch the leash unless the dog became tangled. The researcher also recorded the walk using an i-Phone 7  from 10 m behind. At the end of each walk, participants completed an exit questionnaire  containin = 368/370) videos were coded in their entirety with behavior observation software  were unavailable due to technical problems. Canine behaviors  leash tension files were lost, which we believe to be because the metal handle and case used to contain the components blocked the signal in a certain orientation. This left 339 (n = 339/370) files for analysis . The start and end of each file were determined by matching the timestamps of video and the leash tension file and also by matching three signal peaks at the beginning of the walk with the three repeated “1, 2, 3” verbal cues counted by the volunteer. Data were interpolated in order to make the sample times evenly spaced. Tension was tarred by deducting the minimal value, which visually equaled the baseline value when the device was not connected to the dog, from all measured values. Peak and average tensions over the walk were calculated . Additionally, the directional signal of the accelerometer during the sample immediately prior to the start of a “pull event” was used to determine the pulling direction.max), maximal tension by dog (DTmax) and handler (HTmax) were defined as the maximal tension throughout the walk, recorded for the dog and handler, respectively. Mean tension was calculated by averaging all tension peaks above the threshold. Net mean tension (NTmean), mean tension by dog (DTmean) and mean tension by handler (HTmean) were defined as the mean tension throughout the walk, recorded for the dog and handler, respectively. Dog pulling frequency (DPF) and handler pulling frequency (HPF) were calculated by dividing the number of pulling events caused by the dog and the handler, respectively, by the total walking time.Net maximal tension  was used to assess whether the model improved by entering or removing variables.The 370 interactions were not independent, as dogs were assigned to participants according to the training levels due to safety and welfare considerations. Also, dogs that had been staying in the shelter longer during the research period tended to be walked more often. Generalized linear mixed models were used for analysis to address the repeated measurements. To reduce the numbers of predictors, a bivariate generalized linear model was used to analyze each combination of outcome variables  and predictors . In the analysis of the exit questionnaire, apart from the above predictors, canine and human behaviors, HTOutcome variables were manually transformed for statistical analyses to meet the following assumptions of generalized linear mixed models: (1) residual normality (assessed by observation of quantile–quantile plots), (2) normality of the random effects (assessed by observation of quantile–quantile plots), and (3) homogeneity of variance of residuals . The assumption of no collinearity between covariates was evaluated from variance inflation factors   females and 53 (47.7%) males, all gonadectomized , maximal tension created by the dog (p = 0.014) and maximal (p = 0.0293) and mean (p = 0.0425) tension created by the human. Extraversion was positively correlated with maximal net tension (p = 0.036), maximal tension created by the dog (p = 0.021) and maximal (p = 0.041) tension created by the human. Openness was positively correlated (p = 0.0031), and agreeableness negatively correlated (p = 0.044) with dog pulling frequency. Conscientiousness was negatively correlated with human's pulling frequency (p = 0.039)  .p = 0.027) and high-pitched voice (p < 0.001). The openness score was negatively correlated with the frequencies of total verbal cues (p = 0.0039), attention seeking (p < 0.001), praise (p < 0.001) and high-pitched voice (p = 0.016). The agreeableness score was positively correlated with the frequency of attention seeking (p = 0.04). Finally, the conscientiousness score was negatively correlated with frequencies of communication (p < 0.001) and high-pitched voice (p = 0.035) (The extraversion score was positively correlated with the frequencies of praise (= 0.035) .p = 0.0094), extraversion (p = 0.038) and agreeableness (p < 0.001) interacted with dogs using a higher frequency of total body language. Specifically, a high frequency of hand gestures was observed when the participant scored higher on neuroticism (p = 0.016) and agreeableness (p = 0.0083); a high frequency of physical contact was seen in people scoring high on neuroticism (p = 0.0078), agreeableness (p = 0.0025) and conscientiousness (p = 0.0091)  . Finallyp < 0.001). A high percentage of time spent tail wagging (p = 0.047) and a high frequency of shaking (p = 0.0017) were observed in dogs while interacting with more extravert volunteers. When interacting with volunteers scoring high on openness, dogs presented a high frequency of lip-licking (p = 0.012) but lower percentage of time spent sniffing (p = 0.017). A high score for volunteers' agreeableness was associated with dogs with a high frequency of gaze (p = 0.0077) and lip-licking (p = 0.023) (p = 0.039) and extraversion (p = 0.0079) were more likely to rate higher on factor D (The score of neuroticism was positively correlated with the frequency of dogs' shaking (= 0.023) . Finallyfactor D .p = 0.035) and commands (p < 0.001). However, net mean tension of the leash was negatively associated with the frequency of total verbal cues (p = 0.027), use of a high-pitched voice (p = 0.017) and commands (p = 0.0011)  .p = 0.046) while the frequency of physical contact was positively related to the Factor H score (p = 0.038). The mean leash tension created by dog was negatively associated with the factor H (p = 0.0066) and factor D (p = 0.011) score  score .Volunteers scoring high on neuroticism pulled the leash harder during the walk, which may be related to the frequent body language used by more neurotic people (Kis et al., When walking with people scoring high on neuroticism, dogs licked their lips and shook their body more frequently, which may be a potential indicator of stress (Kogan et al., Volunteers' extraversion was positively correlated with the leash tension from the human, in line with the more assertive and disciplinary parental style observed in more extraverted mothers (Clark et al., Dogs increased maximal leash tension and kept their tails high more often when walking with extravert volunteers, potentially because dogs were aroused by the excited human praise and high-pitched voice, and were more engaged in the interaction with volunteers (McGowan et al., Owners with increased openness were reported to use more gestural and verbal cues when asking their dogs to sit (Kis et al., More lip-licking was observed in dogs walked by more open volunteers, which may be considered as a similar outcome, an appeasement behavior, as interacting with more neurotic volunteers. However, since open volunteers verbally interacted with dogs less frequently, and dogs spent less time sniffing, an indicator of avoiding an uncomfortable conflict situation Cohen, , the higAgreeableness refers to the characteristic and tendency of compliance, cooperation, altruism (Costa et al., Conscientious mothers are more responsive to their children by being more sensitive to children needs, providing support and comfort and following their lead (Clark et al., Cimarelli et al. found that owners scoring high on conscientiousness praise and pet their dogs less frequently (Cimarelli et al., Although extravert owners are more likely to return dogs to the shelter post-adoption Walker, , owners Volunteers talked to dogs more frequently, using more commands, when the net maximal tension was higher, probably because there was greater need for verbal commands when there were strong and sudden pulls. However, the opposite results were observed with fewer high-pitched voices and commands when the net mean tension was higher. In the scenario of higher mean tension, dogs were generally more determined and pulling, resulting in a lack of interaction between dogs and handlers.Volunteers perceived the dog as less obedient and less supportive when they more frequently corrected the dog with negative verbal cues. However, volunteers were more satisfied with the interaction when there was more frequent physical contact between the dog and the human (Protopopova and Wynne, A limitation of this study was that dogs were not randomly matched with participants. Due to concerns for animal welfare and human safety, dogs were assigned to participants based on canine behavior and participants' experiences. Also, the cultural backgrounds of participated volunteers were unclear, which might potentially influence their interaction with dogs Hood, . NeverthThis research provides a qualitative description of associations between human personality and the behavioral dyads that exist between volunteers and shelter dogs when walking on a leash. Neurotic volunteers pulled the leash harder during the walk and tended to interact with dogs using more body language; dogs in turn displayed more lip-licking and body shaking. Extroverted volunteers were associated with stronger maximal leash tension at both the human and dog ends, and they praised the dog more, often accompanied with a high-pitched voice. This elicited more tail-wagging and body shaking by the dog. Open volunteers were less likely to verbally attract the attention of dogs, praise the dogs and talk to them with a high-pitched voice; however, dogs were more likely to pull on the leash, accompanied by more lip-licking but less sniffing. Agreeable volunteers liked to verbally attract the attention of dogs and initiate more hand gestures and physical contact, causing the dogs to pull less frequently, often coupled with more gazing and lip-licking behaviors during the walk. Conscientious volunteers were less likely to pull the leash and tended to have more physical contact with dogs, but they did not prefer verbal communication and a high-pitched voice.In this study, dogs were more likely to be rated as well-behaved when walking with neurotic and extroverted volunteers. A more satisfactory human-dog relationship was detected if the owner and dog shared a similar activity level (Curb et al., The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by University of Queensland Science, Low & Negligible Risk Ethics Sub-Committee. Written informed consent to participate in this study was provided by the participants' legal guardian/next of kin. The animal study was reviewed and approved by Animal Ethics Unit, Office of Research Ethics, The University of Queensland.H-YS, CP, MP, NP, LM, and FG: conceptualization. H-YS, CP, FG, MP, NP, and LM: methodology. FG and H-YS: software and writing—original draft preparation. H-YS, FG, and CP: validation, formal analysis, and data curation. H-YS: investigation. MP and CP: resources. H-YS, CP, MP, NP, and LM: writing—review and editing. H-YS and CP: visualization and funding acquisition. CP, MP, NP, and LM: supervision. H-YS, CP, and MP: project administration. All authors contributed to the article and approved the submitted version.MP was employed as the principal scientist by RSPCA, QLD. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.


To allow better diagnosis and management of psychiatric illnesses, the use of easily accessible biomarkers are proposed. Therefore, recognition of some diseases by a set of related pathogenesis biomarkers is a promising approach. The study aims to assess the usefulness of examining oxidative stress (OS) in schizophrenia as a potential biomarker of illness using the commonly used data mining decision tree method. The study group was comprised of 147 participants: 98 patients with schizophrenia (SZ group), and the control group . The patients with schizophrenia were divided into two groups: first-episode schizophrenia  and chronic schizophrenia . The assessment included the following biomarkers in sera of patients: catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase-1 (SOD-1), glutathione reductase (GR), reduced glutathione (GSH), total antioxidant capacity (TAC), ferric reducing ability of plasma (FRAP), advanced glycation end products (AGEs), advanced oxidation protein products (AOPP), dityrosine (DITYR), kynurenine (KYN), N-formylkynurenine (NFK), tryptophan (TRY), total oxidant status (TOS), nitric oxide (NO) and total protein. Maximum accuracy (89.36%) for distinguishing SZ from HC was attained with TOS and GPx (cut-off points: 392.70 and 15.33). For differentiating between FS and CS, the most promising were KYN, AOPP, TAC and NO . To distinguish FS from HC, maximum accuracy was found for GSH and TOS , and in order to distinguish CS from HC, the most promising were GSH and TOS . Using redox biomarkers would be the most promising approach for discriminating patients with schizophrenia from healthy individuals and, in the future, could be used as an add-on marker to diagnose and/or respond to treatment. For many years, psychiatric research has been concerned with finding and implementing clinical practice biomarkers specific for diagnostic units ,2. The cBiomarkers have been seen as a supplement to diagnosis and cannot replace the current use of the interview-based method. One substance cannot reflect the complicated clinical picture of the disease. The complexity of the biological pathways leads to interactions between many processes, and one change entails another. Some abnormalities occur only due to the functionality formed in the previous step ,6. The pThis study aims to assess the usefulness of examining redox biomarkers (RB) disruption in schizophrenia as a potential biomarker of illness.We recruited 147 participants between 18 and 65 years old. The study population consisted of 98 patients who have schizophrenia (SZ group) diagnosed according to DSM 5 criteria  assessed the severity of psychopathological symptoms using the Positive and Negative Symptom Scale (PANSS) in Polish adaptation . The equWe analyzed the obtained results using Statistica 13 software . The Shapiro–Wilk test indicated the non-Gaussian distribution of most examined factors. As a result, we used non-parametric statistics to characterize participants enrolled in the study. To assess the differences in socio-demographic data between groups, the Man–Whitney U test was applied.To find a potential panel of biomarkers of SZ for both first-episode and chronic SZ, we applied the classification tree model method (C&RT). C&RT is the most often used method for exploring data. The data-mining analysis allows for determining the roles of examined factors and their hierarchy by categorizing patient subgroups. C&RT divides analyzed data into two subsets in the variable hierarchy order, creating a subset tree. Each of the following subsets is more homogenous than the previous. As we mentioned, the C&RT system is elastic and allows for assessing predictors by considering unequal classification costs, which is impossible using other data algorithms.Spearman’s rank correlation coefficients were calculated after they were obtained, and suggested biomarkers related to SZ that could determine the potential effect of treatment and psychopathological symptoms on redox state changes.2, HC 22.8 kg/m2). In the patients’ group, the median length of illness was 24 months. The median value of psychopathological symptoms in the SZ group was 87 (points of PANSS) and was different between first-episode patients (median: 104 points) and chronic patients (median: 72 points). The patients’ groups differed in doses of antipsychotic drugs . Twenty-six patients from the FS group were drug-naive (53.1%).Of the 98 individuals in the SZ group, 47 (46.9%) were female, and of the 49 individuals from the HC group, 26 (55.3%) were female See: . The medThe results of the C&RT analysis are depicted in Firstly, we performed an analysis to distinguish the schizophrenia patients sample from healthy individuals, independent of time since diagnosis. p > 0.05).Maximum accuracy for schizophrenia diagnosis was attained with two of sixteen candidate biomarkers: TOS and GPx. The cut-off point TOS was >392.70, and GPx was >15.33 for schizophrenia see: . When TOThe analysis singled out four factors for discriminating first-episode vs. chronic patients: KYN, AOPP, TAC and NO see: . The cutp < 0.05); KYN , TAC  and NO . There was also a connection between PANSS total score and KYN  and TAC .Due to some differences in PANSS scale scores and equivalents of antipsychotic medication between CS and FE, we decided to assess the effect of these variables on our results. We found a relationship between olanzapine equivalents and AOPP .For discriminating chronic patients from healthy individuals, GSH and TOS were also the most accurate. The cut-off point for GSH was below or equal to 0.26 with 87.5% accuracy, and adding TOS with the cut-off >343.28 raised the accuracy to 100% see: . The levThe aim of this study was to determine potential biomarkers from the set of redox factors and their cut-off points to distinguish individuals with SZ. Recent research has been focused on multiple possible biomarkers to diagnose different diseases for improving sensitivity and specificity ,19,20. IRapid diagnosis of schizophrenia is still a considerable challenge, and none of the blood factors are enough to be identified as an additional biomarker of SZ. There is an urgent need to find biologically accessible, peripheral biomarkers. Several potential candidates for them have been reported . An incrIn data analysis, the traditional statistical approaches have limited utility in classification . These mA large amount of evidence indicates an important role of pro/antioxidant disturbance in individuals with schizophrenia ,10,24. HOur analysis showed that the most promising factors in distinguishing schizophrenia patients were TOS and GPx, with 98.81% accuracy. There is a relationship between TOS and negative symptoms, but not all studies supported that TOS is higher in schizophrenia patients . It has 2O2 Equiv./L [Kulaksizoglu and Kulaksizoglu have determined the cut-off point for TOS as 5.9 . HoweverEquiv./L . AccordiEquiv./L .Some studies indicate lower Gpx is related to SZ pathophysiology and a worse course of disease . In our Gpx is crucial in the GSH system, in which changes were observed in chronic and FS patients in our study. The GSH is a non-enzymatic antioxidant that exists at high intracellular levels. The GSH system can counteract the excitotoxicity effect of glutamate and unfavourable oxidative changes in the CNS. The higher brain GSH was accompanied by the faster response to antipsychotics treatment in the drug-naïve first-episode group, and higher levels of GSH reversed the negative impact of hyperglutamatergic state in patients with schizophrenia ,35. The Blood GSH can cross the blood–brain barrier. Thus, plasma or serum GSH may reflect central GSH. Some authors have also suggested that there might be a relationship between NMDA and GSH. GSH deficits may induce NMDA receptor hypofunction. On the other hand, synaptic NMDA receptor activity boosts and manages GSH metabolism ,39. In tIn comparing the first episode to chronic schizophrenia, we found four candidates: KYN, AOPP, TAC and NO. We proved lower KYN was related to a chronic state, inconsistent with meta-analysis from 2021 . In the p < 0.05 for antipsychotic equivalents and R = 0.38; p < 0.05 for PANSS) and TAC . Inverse relationships  suggest KYN and TAC could be considered markers related to psychopathology and/or treatment response. However, the results need further investigation.In our analysis, PANSS scale scores and equivalents of antipsychotic medication were related to RB biomarkers, especially KYN . In one study, higher levels correlated with clinical severity scores in SZ .Similar to our results, in a study assessing TAC in chronic and FS patients, this variable did not differentiate them from healthy individuals. The authors also did not show a difference dependent on the duration of the illness, but the study group was relatively small (42 SZ and 42 HC) . InteresThe studies that assessed NO concentration in SZ are inconsistent ,48. The It is possible that only some redox balance factors are related to the pathophysiology of psychiatric disorders. In our study, many RB  were not related to schizophrenia. On the other hand, medication (as a tool to reduce disruption homeostasis linked with illness) could affect these factors’ levels in the blood. However, there are many fundamental issues with no answer in terms of biomarkers of schizophrenia, and which need clarification.The limitations of our study must be mentioned. Firstly, many changes in the oxidative system are not specific to psychiatric diseases and have been reported to be involved in the pathogenesis of various  chronic diseases ,53. The In conclusion, we find that using RB would be a promising approach to differentiating patients with schizophrenia from healthy controls and, in the future, could be used as an add-on marker to diagnose and/or respond to treatment. Confirmatory studies with a larger sample size in different stages of illness should be performed to establish utility redox balance-related factors as a biomarker of SZ.


Adipose tissue is considered an endocrine organ, and its excess compromises the immune response and metabolism of hormones and nutrients. Furthermore, the accumulation of visceral fat helps to increase the synthesis of cortisol. The hypothalamus-pituitary-adrenal (HPA) axis is a neuroendocrine system involved in maintaining homeostasis in humans under physiological conditions and stress, and cortisol is the main hormone of the HPA axis. It is known that a stress-induced diet and cortisol reactivity to acute stress factors may be related to dietary behavior. In obesity, to reduce visceral adipose tissue, caloric restriction is a valid strategy. In light of this fact, the aim of this study was to assess the effects of a commercial dietary ketosis program for weight loss on the sympathetic nervous system and HPA axis, through evaluation of salivary cortisol and GSR levels. Thirty obese subjects were recruited and assessed before and after 8 weeks of Very Low Calorie Ketogenic Diet (VLCKD) intervention to evaluate body composition and biochemical parameters. Salivary cortisol levels and GSR significantly decreased after dietary treatment; in addition, body composition and biochemical features were ameliorated. The VLCKD had a short-term positive effect on the SNS and HPA axes regulating salivary cortisol levels. Finally, the effects of the VLCKD on the SNS and HPA axis may lead to more individualized treatment strategies that integrate obesity and stress and support the usefulness of such therapeutic interventions in promoting the reduction of the individual disease burden. The center of stress management consists of the sympathetic nervous system (SNS) and the hypothalamic-pituitary-adrenal axis (HPA). These systems coordinate the response of the various physiological systems, leading the body to equilibrium. There is robust evidence from animal and a human study that stress causes coincidental activation of nerve and system cells and also the unharnessing of many biologically active compounds, as well as catecholamines and glucocorticoids ,2. DysreCortisol receptors, present in most cells in the body, receive and use the hormone in different ways. For example, when the body is on alert, cortisol can alter or disrupt the functions that stand in the way . These cThirty male subjects, affected by obesity and aged between 50 and 60 years (±8 years), were enrolled by the Laboratory of Physiology, Department of Clinical and Experimental Medicine, University of Foggia. The study was approved by the local Ethics Committee 22 May 2018, n◦440/DS, and conducted according to the ethical principles of the Declaration of Helsinki. Written informed consent was obtained from all participants. The sample chosen was completely male because, given the age range of our population (50–60 years), we preferred to consider only a male population to minimize hormonal interference due to menopause, which could have affected a group of women of the same age. In our population, we evaluated body composition by measuring height, weight, body mass index (BMI), and visceral adipose tissue (VAT) using dual-energy X-ray absorptiometry (DEXA). Serum samples were collected after a 12-h overnight fasting period. Serum aliquots were stored at −80 °C. As previously reported, for all participants, glucose, total cholesterol, low-density lipoprotein (LDL), high-density lipoprotein (HDL), triglycerides, LDH, aspartate transaminase (AST), alanine transaminase (ALT), IL-10, TNF-a, adiponectin, and Orexin-A were measured ,22.All participants underwent a general medical examination . To veriObese subjects followed a VLCKD, as per an advertised weight loss program , consisting of <50 g/d sugar from vegetables, 43% fat, 43% protein, 14% carbohydrates, and 700–900 kcal. The amount of protein ranged between 1.0 and 1.2 g per kilo of ideal weight. Although, the dietary intervention profile consisted of three totally different stages, for the needs of this study, only active ketogenic phases of the primary stage were considered. This stage had an 8-week length, to permit participants to realize some of their weight loss target. In this period, vitamins, minerals, and polyunsaturated fatty acid were provided in accordance with international recommendations.g for 15 min at 4 °C. To gauge the absence of blood contamination, a secretion blood contamination kit was used . Corticoid concentrations were measured using commercial kits , as per the manufacturers’ directions. All samples were tested in triplicate and analyzed in duplicate.Salivary samples were collected between 09:00 and 11:00 before the VLCKD intervention and after eight weeks of VLCKD intervention using of cotton swabs . Participants were asked to position the cotton swab in their mouth for a minimum of two minutes and then insert it into a special plastic tube. Samples were sent as soon as possible to the laboratory and kept at −20 °C until the corticoid assay. The secretion samples were centrifuged at 1500× Within five minutes following spittle collection, electrodermal response information was recorded. The electrodermal response parameters were measured with a SenseWear professional Armband™ , which was worn on the appropriate arm over the striated muscle at the center between the appendage and olecranon process, as suggested by the manufacturer .p-value ≤ 0.05 was used for statistical significance. Secretion of corticoid, adiponectin humor concentrations, and total cholesterol were related to Pearson’s tests, as per the information distribution. A p-value < 0.05 was considered statistically significant.We performed applied mathematical analyses using StatView software 5.0.1.0 . All information is given as means ± SE; a two-tailed paired t test was used to check for applied mathematical significance of the outcome variables. A t-test analysis showed that glycemic, lipid profile, and inflammatory status were strongly ameliorated in these subjects after the diet. In p-value < 0.05. In the VLCKD patients after diet intervention, there was a large change in anthropometric and biochemical parameters. Many parameters such as weight and BMI were statistically reduced in VLCKD obese subjects after the diet. Furthermore, p < 0.05) and cortisol (1.31 μmol/L vs. 1.01 μmol/L p < 0.01). Salivary cortisol levels in the VLCKD obese patients were strongly decreased after the VLCKD intervention  . In addiOur data provide further support for a possible effect of cortisol reactivity on food intake in people with obesity, but not in healthy weight controls. Therefore, the physiological mechanism of stress-induced nutrition is a complex interaction between many different hormones, whose secretion and activity are affected by glucocorticoids ,39,40,41In addition, we also found a reduction in GSR, which confirms the role of stress and the reactivity of cortisol in food intake, confirming the association between the SNS and the HPA axis ,38,39. A VLCKD has a short-term positive effect on the SNS and HPA axes. In addition, there is a strong relation between obesity and stress, and cortisol has a key role. For these reasons, VLCKD has an important role reducing stress and regulating salivary cortisol levels. Finally, the effects of VLCKD on the SNS and HPA axis may lead to more individualized treatment strategies that integrate obesity and stress in support of the reduction of the individual disease burden.Limitation of the study: this is one of the first studies investigating VLCKD effects on the SNS and HPA axis; however, a limitation to this study is the small number of male patients considered. For this reason, it is our intention to expand the series to include the female population in further studies on this subject.


Early intervention programs positively affect key behaviors for children with autism spectrum disorder (ASD). However, most of these programs do not target children with severe autistic symptomatology associated with intellectual disability (ID). This study aimed to investigate the psychological and clinical outcomes of children with severe autism and ID enrolled in the Tailored and Inclusive Program for Autism—Tours (TIPA-T). The first step of the TIPA-T is the Exchange and Development Therapy (EDT): an individual neurofunctional intervention consisting of one-to-one exchanges between a child and a therapist taking place in a pared-down environment. It aims to rehabilitate psychophysiological abilities at the roots of social communication through structured sequences of “social play.” Cognitive and socio-emotional skills and general development were evaluated with the Social Cognitive Evaluation Battery scale and the Brunet–Lézine Scale—Revised, respectively, before and after 9 months of intervention in 32 children with ASD and ID. Autistic symptomatology was evaluated with the Behavior Summarized Evaluation—Revised scale at five time-points in a subset of 14 children, both in individual and group settings. Statistically significant post-intervention improvements were found in cognitive and socio-emotional skills. All but one child showed improvements in at least one social domain, and 78% of children gained one level in at least four social domains. Twenty-nine children improved in cognitive domains, with 66% of children improving in at least three cognitive domains. Autistic symptomatology evaluated in one-to-one settings significantly decreased with therapy; this reduction was observed in more than 85% of children. In group settings, autistic symptomatology also decreased in more than 60% of children. Global developmental age significantly increased by 3.8 months. The TIPA-T, including EDT in particular, improves socio-emotional skills of most children with ASD and reduces autistic symptomatology, yet with heterogeneous outcomes profiles, in line with the strong heterogeneity of profiles observed in ASD. At the group level, this study highlights the benefits of the TIPA-T for children with severe autism and associated ID. Assessment of autistic core symptoms showed an improvement of social interaction, both in one-to-one and group evaluations, demonstrating the generalizability of the skills learned during the EDT. As defined in the Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition , autism Early intervention for children with ASD has been recognized as a health and educational priority , 7. EarlExisting intervention programs mostly target children with no or mild intellectual deficiency. The TIPA-T program evaluated in the current study is dedicated to all children, from toddlers to young adults, including those with severe autism and associated intellectual disability.The TIPA-T is a tailored and global program based on functional, developmental, and multidisciplinary assessments of the children. The program is set up by a multidisciplinary team, including psychiatrists, psychologists, speech therapists, psychomotor therapists, social workers, teachers, and nurses located in the child psychiatry intervention units in the Center of Excellence for Autism in Tours (EXAC-T) (France). The program is tailored to each child's age and needs following an integrative approach to the treatment of ASD and includes both individual and collective sessions. The current paper focuses on the early phase of TIPA-T, dedicated to children between 2 and 6 years old. The weekly program duration is 20 to 25 h following the recommendations of the High Authority for Health in France , integrating individual and collective care times  and school sessions in mainstream kindergarten with individualized support.Collective interventions, spread on the whole day, comprise speech and psychomotor activities performed by trained professionals, an educative program proposed by nurses and specialized teachers within the child psychiatric unit, alongside group free play aiming at working on socialization, communication, and autonomy. Collective interventions also integrate inclusive school sessions with individual support.Individual intervention mainly consists of sessions of EDT, an individual neurofunctional therapy based on the experience and practice of a multidisciplinary team –24. The A growing body of evidence on early interventions for children with ASD highlights a large variability in children's response to treatment . HoweveThis study aimed to describe the evolution of key autistic symptoms and behaviors in some children with severe ASD and concurrent ID following a 9-month early intensive intervention program, the TIPA-T, centered on the EDT. We hypothesized that the TIPA-T would yield progress in socio-emotional skills assessed with the SCEB and a reduction of autistic behaviors with the BSE-R.The sample consisted of 32 children  with a diagnosis of ASD, according to International Classification of Diseases 10  and Diagnostic and Statistical Manual of Mental Disorders, Fifth Edition ; mean ± The implementation of the EDT is highly structured by visual cues, visually based schedules, and the implementation of routines. EDT sessions take place two or three times a week and last approximately 20 min; sessions are always adapted to the child's attention and concentration skills.The EDT is based on three general principles: serenity, availability, and reciprocity. The organization of EDT sessions aims at enforcing these general principles. To respect the principle of serenity, the EDT takes place in a bare room, thus creating a very sober space devoid of any distraction and precipitation. Before the session, the therapist prepares the room; that is, they choose furniture that will produce the best environment for inducing interactions with the child  and the toys for the child. The aim is to create an environment that is stable from one session to another. The choice of toys is based on previous experience with the child; toys that have produced high-quality interactions at previous sessions are preferred.To enforce the availability principle, the toys are offered one by one, in a predefined order, to keep the amount of stimulation to a minimum and focus the child's attention to the play at hand; nonetheless, the organization remains flexible to adapt to the child's envy. To keep the child involved in the sessions, both interactive and relaxing activities are proposed. This allows optimizing exchanges between the child and the therapist.Perceptual-motor  and socio-emotional  sequences established around free play aimed at progressively increasing the synchronization between the child and the adult to promote reciprocity by fostering sociability. The therapist is continuously attentive to the child's communicative manifestations, however discreet they may be, to adapt the therapy to the child's reaction, and to include the child's initiatives in the sessions. If a child shows no interest in the proposed activity, the therapist tries gently to bring their attention back to the current activity by soliciting them gently. Importantly, they do so without showing disapproval to not reinforce the child's behaviour by a mark of attention. By doing so, the therapist manages to reengage the child and avoid a situation of failure. This permits that the child does not keep in mind a failed interaction. Finally, the adult promotes role-playing games and gradually introduces variations in the proposed scenari, considering the child's progress, so that established routines are not ritualized.The Brunet–Lézine Scale—Revised  is an adCognitive and socio-emotional skills were assessed with the Social Cognitive Evaluation Battery –42. BaseAutistic symptomatology was assessed using the BSE-R scale , 43. TheScorings of the Brunet–Lezine scale and the SCEB were performed at the beginning of therapy and after 9 months by psychologists experienced with these tools. The initial evaluation necessary to work out a tailored therapy program constitutes the baseline. The BSE-R scale was evaluated both in a one-to-one therapy session, through recordings of EDT sessions, and in collective settings to study a generalization of acquired skills at months 1, 3, 5, 7, and 9 of therapy.Statistical analysis and figures were made with R  within * interquartile range (IQR) Q3 + 3 * IQR]; in cognitive domains after therapy} using the IQR method: his averaged SCEB scores were all outside the range (Q1 - 1.5 * IQR Q3 + 1.5 * IQR) ; before removing participant 28, data in the cognitive domain after therapy did not follow a normal distribution (p = 0.01).SCEB scores averaged across all social and all non-social domains  were anaT-tests for fixed effects parameters use the Satterthwaite's method for the degree of freedom calculation as in lmerTest  for the population and two random effects for each subject. Random effects for a particular subject were the deviations in intercept and slope of that subject's time trend from the population. The model allowed for the correlation of random effects for the same subject due to the relationship between intercept and slope . The modlmerTest .T-test after removing one extreme outlier ; effect size was computed as Cohen's d. DAs for each area were analyzed with Wilcoxon signed-ranked test on paired data. Participant 32 was identified as an extreme outlier in SDA, showing an improvement of 19 months, and was therefore removed for SDA analysis.Developmental assessments using Brunet–Lézine data were analyzed at the beginning and at the end of therapy. GDA was analyzed using a paired Student  = 125.89, p < 0.001, η2[g] = 0.32] and domains  = 0.26] on the average levels of the SCEB but no interaction  <0.001;  = 129.24, p < 0.001, η2[g] = 0.27], area  = 0.21], and no interaction.A repeated measure ANOVA with time and domain as within-subject factor revealed main effects of time  and cognitive domains . After 9 months of therapy, mean levels had increased by approximately 0.5 for both social  and cognitive  domains . To haveExploration of individual data  revealedAll but one child showed improvements in at least one social domain, and 78% of children gained one level in at least four social domains. Twenty-nine children improved in cognitive domains, with 66% of children improving in at least three cognitive domains.Exploration of the interaction and modulation deficits assessed with the BSE-R in individual EDT session revealed an average reduction of symptoms of 0.92 and 0.88, respectively. Attenuation of interaction deficits was observed in 93% of children, and modulation deficits decreased in 86% of children.(14) = 22.3; p < 0.0001} and slope , highlighting that interaction deficits before therapy were high but decreased by approximately 0.1 point  per month of therapy  for time, across the population, were significant for both intercepts {β [95% confidence interval (CI)]: 3.6 [3.3 3.9]; T therapy . The sta therapy  for fact(14) = 20.2; p < 0.0001] and for slope  and slope  for factor 1 ((14) = 12.8; p < 0.0001] but not for slope . Correlation coefficient for within-subjects' random effects of intercept and slope was high : 4.1 (3.8 4.3); Tfactor 1 , highligfactor 1 . Effect factor 1  evaluateh -0.98; .(30) = 10.15, p < 0.001, Cohen's d = 1.5; GDA assessed with the Brunet–Lézine scale significantly increased by an average of 3.8 months with therapy [GDA, Tr > 0.7): postural developmental age  = 8.49, p < 0.001, Cohen's d = 1.5, SDA: V = 465, p < 0.001, n = 32, r = 0.87.Keeping outliers in the statistical analysis does not change observed effects: GDA: TUsing clinical tools tailored for ASD and sensible to subtle changes occurring over a short amount of time (the SCEB and the BSE-R), positive outcomes were observed in children with severe ASD and concurrent ID following a 9-month early intensive intervention program, the TIPA-T centered on the EDT in a specialized medical center. The TIPA-T, including its pivotal first step, the EDT, allowed for a general improvement for both cognitive and socio-emotional domains and a reduction in the severity of autistic symptomatology. In addition, assessment of autistic core symptoms with the BSE-R showed a decrease of interaction deficits, in particular, both in one-to-one and in group evaluations. The latter demonstrates the generalizability of the skills learned during the EDT.Children's developmental trajectories were positive, but their evolution was atypical and characterized by uneven progress. Large individual variations in outcomes were observed in this study, consistent with the findings from previous research , 24, 60 Results of this study are consistent with other follow-up studies showing that early and intensive intervention program over a relatively short period yields positive outcomes for children with autism. Dawson and collaborators  showed tDifficulties in assessing the benefits of intervention programs arose from the lack of clinical tools dedicated to assessing subtle changes occurring over a small period . Here, wUsing these assessments complemented the findings obtained with a classical tool such as the Brunet–Lézine—Revised scale. Results from the Brunet–Lézine—Revised scale showed a general improvement in DA, as well as domain-specific improvement. On average, the 9-month therapy yielded a 3-month gain in DA, and importantly, this increase in DA was observed in a large majority of children. This highlights the benefit of the TIPA-T, including specifically the EDT for the cognitive development of children considering that children with low IQ tend to maintain their IQ over time , consistThere are several important clinical implications for these data. When diagnosing young children with ASD, it is important to assess cognitive skills and social-reciprocal interaction deficits and abilities using standardized tests. Both measures are strongly related to outcomes and are correlated with each other. These measures can help clinicians to assess responsiveness to intervention and treatment planning. In addition, early social-interaction abilities may be a pivotal skill that should be addressed in intervention programs. The study also emphasizes the effectiveness of the intervention at a very early age across the autistic symptomatology severity range. A therapy such as the EDT might also benefit from being developed in at-home settings by providing support and teaching parents the functional basis and the implementation of EDT at home.The TIPA-T, including specifically EDT, improves cognitive and social skills and core symptoms of autism of most children with severe ASD and associated ID. Assessment of autistic core symptoms with the BSE-R showed a decrease of interaction disorder in particular, both in one-to-one and group evaluations. Importantly, this demonstrates the generalizability of the skills learned during the EDT. However, as for other intervention programs in autism, large individual variations were seen in the outcomes, in line with the strong heterogeneity of profiles observed in ASD. Data acquired in the current study do not allow understanding the reasons for the therapy being extremely successful in some children and less in others. Future studies should aim at identifying predictive factors of success to potentiate positive outcomes.The raw data supporting the conclusions of this article will be made available by the authors, without undue reservation.The studies involving human participants were reviewed and approved by CHRU Tours. Written informed consent to participate in this study was provided by the participant's legal guardian/next of kin.CB, J-LA, and RB contributed to conception and design of the study. RB, MG, JM, PD, and AR contributed to data collection. SR organized the database. ML performed statistical analyses. RB, ML, JM, FB-B, and J-LA wrote the first draft of the manuscript. All authors contributed to manuscript revision, read, and approved the submitted version.J-LA is the author of the SCEB edited in the Pearson France-ECPA. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest.All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article, or claim that may be made by its manufacturer, is not guaranteed or endorsed by the publisher.


Neoregelia cruenta during two years (eight seasons). Significant differences among areas were found in some seasons for total nematode abundance and/or the abundance of some trophic groups, but no pattern emerged relative to the level of AD. Anthropic disturbance did not impact nematode trophic structure possibly because the water PCPs remained fairly similar in all three areas. Our results do not corroborate previous reports that AD alters phytotelma water. On the other hand, our findings support previous studies suggesting that nematodes inhabiting bromeliad phytotelma are not good candidates for AD-bioindicators.Phytotelmata (sing. phytotelma) are plant-associated reservoirs of rainwater and organic debris. These freshwater ecosystems are found in tree and bamboo holes, pitcher plants, and tank-forming bromeliads. Some studies suggest that anthropic disturbance (AD) may change the physico-chemical properties (PCPs) of the water retained in the phytotelma, and indirectly impact its biota. Hence, new AD-bioindicators could be found in the phytotelma biota. To test this hypothesis, three areas of Atlantic Forest were selected, distinct only by the level of long-term AD. In these areas, we monitored the nematode trophic structure and the water PCPs in the bromeliad  Anthropic disturbance (AD) is a major threat to forest ecosystems, primarily through unmanaged wood harvesting, deforestation, and fragmentation. Microorganisms, micro- and mesofauna have been intensively studied as bioindicators of AD and post-AD restoration. Most studies have focused on benthic and soil communities and ecotoxicology assessments .Nematodes have been widely used as bioindicators . In terrNematodes have been known to inhabit phytotelmata since the 1920s, but only recently have ecologists investigated them in tree holes , plasticThere is no clear understanding of which environmental factors impact phytotelma nematodes. The amount of organic debris impounded in the phytotelma and the biomass of algae living in the phytotelma water correlate positively with nematode abundance and diversity. Fluctuations in rainfall do not impact the nematodes, while the mean air temperature may affect nematodes in temperate regions, but not in tropical ones  to impact the inhabiting biota. Indeed, these water PCPs have been found to impact communities of algae, archaea, bacteria, micro- and macroinvertebrates . NonetheAlthough natural changes in the water PCPs have no impact on the abundance of phytotelma nematodes, AD may change this pattern. Anthropic disturbance is known to reduce bromeliad species richness and diversity . Wood haHence, we hypothesized that AD might change the PCPs of the water retained in the bromeliad tanks; and that such changes might impact the nematode trophic structure. Corroboration of this hypothesis would mean that nematodes dwelling in tank-forming bromeliads could be useful bioindicators of AD or post-AD restoration in bromeliad-rich ecosystems.To test this hypothesis, we investigated bromeliad-rich areas of the Atlantic Forest biome that have been submitted to distinct degrees of AD. In these areas, we monitored the water PCPs and the nematode trophic structure during a two-year period, and sought correlations between those parameters and the level of AD.http://www.icmbio.gov.br/parnajurubatiba/), in the state of Rio de Janeiro, Brazil. The park protects a large area of restinga, an Atlantic Forest ecosystem that includes beaches, dunes, lagoons and areas subject to periodic flooding, in which herbaceous and shrubby species predominate. The region’s climate is AW  according to the Köppen classification, with an average annual temperature of 23°C and a yearly rainfall ranging from 1,000 to 1,350 mm.The study was conducted in Restinga de Jurubatiba National Park (RJNP)  L. B. Smith, which thrives abundantly in the RJNP, shaded by short trees and shrubs. Only mature plants with inflorescence were sampled because this phenological stage is known to support a more abundant nematode community ; pH; dissolved oxygen ; dissolved solids ; and electrical conductivity . These were measured in the field with an Icel Manaus® meter (model PH–1500) with appropriate sensors. A 10 mL aliquot was collected in a new, sterile penicillin flask and used for measuring dissolved organic carbon  and dissolved nitrogen , using a Shimadzu meter (model TOC–Vcph). Macroscopic organic debris (OD) fallen into the phytotelma was collected in a paper bag, dried for 24 hr in an oven at 80°C, and expressed in grams.In areas 1, 2, and 3, samplings were carried out from June 2014 through March 2016, once every season, for a total of eight sampling dates. On each sampling date, in each area, eight bromeliads were chosen along a random path. This process was carefully done not to repeat previous paths. The water retained in the bromeliads was suctioned and submitted to the following measurements: volume (mL); temperature . The resulting suspension was submitted to the method of ® microscope with Nomarski interference contrast in the 40 and 100X lenses.The 24 suspensions were reduced to a volume of 5 ml and observed entirely in Peter’s slides for nematode counting. The total abundance of nematodes and the abundance per trophic group were computed. Nematodes were assigned as plant, hyphal, or bacterial feeders, unicellular eukaryote feeders, or carnivores  by examihttps://doi.org/10.6084/m9.figshare.12925571. Unicellular eukaryote feeders were found only occasionally and plant feeders were not found. Hence, no statistics were applied to these groups. Nematode counts – total, hyphal, and bacterial feeders, and carnivores – were tested for homogeneity of variances (Cochran and Bartlett tests) and for normality of errors (Lilliefors test), at 5% probability. Since the assumptions were satisfied, nematode counts were evaluated through a two factor-ANOVA using the R language v.3.6.1 (x| > 3 SD) were considered outliers and removed from the dataset. The pairwise comparison among least square means was done by Tukey test at 5% probability using the R language v.3.6.1. These comparisons were made by pooling data from all eight sampling dates (seasons), and by decomposing the data per season.The assays’ raw data are publicly available at  v.3.6.1 . The linWhen data from the eight sampling dates were pooled and submitted to ANOVA, there was significance for the abundance of hyphal feeders and carnivores in relation to areas . The Tukrestinga. We did not measure canopy coverage over the bromeliads, but we expected that less canopy coverage would lead to less impoundment of organic debris  in the phytotelma. This would negatively affect the biomass of plant litter-decomposing fungi and the abundance of hyphal feeder nematodes.The greater abundance of hyphal feeder nematodes in the most disturbed areas is counterintuitive. Particularly in area 1, the bromeliads had less coverage by trees and shrubs due to wood harvesting and fragmentation of the N. cruenta.The ANOVA of data from the eight sampling dates also revealed significance for nematode abundance  in relation to seasons, and the area x season interaction . When weN. cruenta .On the other hand, at times other unknown factor(s) impacted nematode abundance differently in the areas 1, 2, and 3. Major climate variables such as air temperature, rainfall, sun, and wind exposure might affect bromeliads – and their phytotelma – differently depending on the level of AD. Nonetheless, air temperature and rainfall had no impact on the nematode trophic structure in the phytotelma of Anthropic disturbance did not impact the nematode trophic structure possibly because it did not affect the phytotelma considerably. Although areas 1, 2, and 3 are distinct in their long-lasting levels of AD, the two-year monitoring of the water PCPs revealed similar values in all parameters , with noRecently, restinga areas did not cause changes in the water PCPs. Also, AD did not affect the nematode trophic structure. Hence, it appears that the trophic structure of nematodes dwelling in bromeliad phytotelma is not a useful bioindicator of AD.Our study is the first to test the hypothesis that phytotelma fauna can be a useful bioindicator of AD. The long-lasting AD affecting the The possibility remains that AD impacts the abundance of particular nematode functional guilds, genera or species in bromeliad phytotelma. Also, higher levels of AD than those observed in areas 1 and 2 might affect the bromeliads to the extent of impacting the phytotelma nematodes. On the other hand, much needed studies on nematodes inhabiting tree and bamboo holes and pitcher plants might reveal a higher impact of AD on those nematodes, and their usefulness as bioindicators.