The purpose of the present study was to examine the effectiveness of the injury awareness and prevention programme P.A.R.T.Y. (Prevent Alcohol and Risk-Related Trauma in Youth) in Germany. On a designated P.A R.T.Y. day, school classes spend a day in a trauma hospital experiencing the various wards through which a seriously injured person goes. A further goal of the study was to reveal indications of the programme’s mechanism of action by testing theory-based impact models of fear appeals and cognitive beliefs.n = 330), as well as pupils who did not attend the programme (n = 244), were interviewed with a standardised questionnaire. They reported risk behaviour, feelings of threat and cognitive beliefs about road traffic. The data were analysed using a meta-analytical approach to estimate an average effect size across the different P.A.R.T.Y. days. Path models were used to identify possible mechanisms of action.In a quasi-experimental longitudinal study with three measurement times the participants of 19 P.A.R.T.Y. days  could be identified. But for these predictors no or only short-term effects were observed in the meta-analysis.Fear appeals as used primarily in the P.A.R.T.Y. programme appear to cause behavioural changes only to a limited extent and only in the short-term, especially if the strengthening of psychosocial resources is not given sufficient consideration. The participants must also cognitively process the experiences in the hospital. Accordingly, consideration should be given to how the P.A.R.T.Y. program could be adapted to complement the fear appeal with cognitive components.The online version contains supplementary material available at 10.1186/s12889-022-12918-2. Injuries are the most common cause of death among teenagers and young adults worldwide. Road traffic accidents are a significant factor for the high incidence of injuries , 2. PrevThe prevention programme P.A.R.T.Y. (Prevent Alcohol and Risk Related Trauma in Youth) is another example of such an intervention. It is a one-day injury awareness and prevention program for youth. A school class spends a day in a trauma centre and experiences the various wards of a seriously injured patient. The P.A.R.T.Y. programme was developed in Canada and has been carried out there for more than 30 years. The programme development and implementation was motivated by the fact that injuries are the leading cause of death in young people and that most of these injuries are predictable as well as preventable .Hence, it was also introduced in many other countries, such as Australia, New Zealand and the United States. The German Society for Traumatology and the Academy of Traumatology GmbH have adapted the concept to German standards. Since 2012, it has been implemented in over 40 trauma centres in Germany. The objective of the programme is to change the behaviour of young people by illustrating the negative consequences of traffic accidents. It is intended to increase awareness for serious consequences of risk behaviour and aims to reduce accidental injuries and deaths in this age group. It is based on learning by means of real people’s experiences through vividness and emotions .Initially, the participants receive two half-hour lectures, each on the topics of trauma and prevention, usually held by a trauma surgeon (trauma) and a police officer (prevention). Afterwards, the pupils pass through the individual wards that a severely injured patient is likely to go through. These are the ambulance, the emergency ambulance/trauma room, the intensive care unit and the normal ward. A so-called P.A.R.T.Y. guide – a hospital staff from the medical or nursing service – accompanies the pupils. The participants get an idea of how a seriously injured patient is cared for in each ward. If possible, they can talk to the patients there. Afterwards, the participants visit physiotherapy and see how tedious and difficult rehabilitation can be after a serious injury. Finally, there is a 20-min talk with a former seriously injured patient. The P.A.R.T.Y. day ends with a joint reflection on the day.Previous evaluation studies find short-term programme effects (1–2 weeks after implementation) on knowledge and attitudes –21 as weThe contents of P.A.R.T.Y. indicate that the programme attempts to change the behaviour of young people, particularly through fear appeals. Whereas most practitioners seem to judge fear appeals as very effective for changing behaviour, researchers are more sceptical. This scepticism seems appropriate, because after all, some meta-analyses support this attitude , 26. HowThe debate presented thus shows that the effectiveness of fear appeals may depend on certain framework conditions and circumstances . FinallyTherefore, the first objective of this study was to determine the effectiveness of the fear appeals-based P.A.R.T.Y. programme in Germany for changing risk behaviour in traffic. The second objective was to investigate the impact mechanism by which the behaviour change is supposed to be achieved. Neither the original programme description nor the previous evaluation studies provide a theory-based impact model that describes the process of behaviour change. Especially the effect of fear appeal is of interest, as participants of the programme are supposed to be convinced by negative consequences of traffic accidents.So far, there is no programme theory that explains the intended process of behaviour change underlying the P.A.R.T.Y. programme. However, a well-formulated theory is necessary to understand the effectiveness of a prevention programme and to further improve it . From thFear appeals aim at motivating people to omit disapproved behaviour or to show other approved behaviour by conveying negative or painful consequences of this disapproved behaviour . In roadDespite intensive research on the effectiveness of fear appeals over the last few decades, the presented debate shows that messages intended to generate emotions such as feelings of fear and threat are not easy to handle. Findings suggest that fear appeals can activate opposing mechanisms, resulting in an inconsistent picture of the effectiveness of fear appeals –41. WittWithin the P.A.R.T.Y. programme information about types of injury, their consequences or care processes are conveyed to the participants. This information may also change the participant’s behaviour. The Theory of Planned Behaviour (TBP) developed by Ajzen describes how cognitive beliefs and the change thereof can influence behaviour . This thP.A.R.T.Y. is a setting-based prevention approach to reach young people in a part of their living environment, namely the school. The program itself is implemented in clinics. Thus, the evaluation study had to be conducted in collaboration with schools and clinics. Furthermore, this is a group-based intervention, since the pupils participate in P.A.R.T.Y. together with their classmates. Therefore, they are considered as members of a cluster (the class) rather than individuals. In the school year 2016/17, a total of 19 P.A.R.T.Y. days were included in cooperation with seven trauma centres and twelve schools in five German federal states .A quasi-experimental study was conducted with three measurement times. Nineteen P.A.R.T.Y. days were examined in seven different trauma centres. Twelve schools took part with 19 intervention classes. Eleven parallel classes served as a control group. A random allocation of classes to an intervention and a control group was intended, but only possible in one school. Likewise, not every school was able to provide a control class. Therefore, there was a simple one-group pre-post-test design without control group for eight of the 19 P.A.R.T.Y. days. These differences in the design were accounted for in the analysis.The surveys were carried out consistently in the classrooms of the participating classes. The questionnaires were usually handed out personally by project staff and then collected again after completing. In this way a high response rate was ensured and the loss of completed questionnaires (e.g. by post) was prevented. In some cases, the survey was carried out by teachers if requested by the school for organizational reasons. The baseline survey (T0) took place immediately before the P.A.R.T.Y. intervention, usually the day before the intervention. The post survey (T1) was usually conducted 1 day after each P.A.R.T.Y. intervention. For the follow-up (T2), participants completed the questionnaire four to 5 months after the P.A.R.T.Y intervention. All methods were approved by an Ethics Committee and performed in accordance with relevant guidelines and regulations. An informed consent was obtained from a parent and/or legal guardian.The questionnaire contained scales measuring the effects of the P.A.R.T.Y. programme, namely self-reported behaviour in traffic. Following the Generic Error Model System –53, itemThe questionnaire was extensively pretested and revised. Table In total 799 pupils took part, 574 of them at all three measurement points. The sample characteristics are shown in Table To determine the effectiveness of the intervention the data analysis was supposed to calculate the average effect size for the intervention groups compared to the control groups. By doing this, two characteristics of the study needed to be taken into account.First, the data obtained from the participants are not independent because they participated as school classes (clusters). The same applies to the participants of the control group. Consequently, the statistical dependency caused by analysing clustered data has to be taken into account. Otherwise, the standard error of the calculated effect sizes is biased. Second, the P.A.R.T.Y. days took place in different clinics and were therefore not a standardized intervention. The implementation of the intervention in the daily hospital procedures, different persons involved in the programme as well as the characteristics of the participating classes are components, which produce an unavoidable heterogeneity of the P.A.R.T.Y. days between the clinics, but also partly within the same clinics. Therefore, based on the data, there is in fact a 19-fold study replication of the P.A.R.T.Y. intervention, which is always similar in structure, although locally differing. Thus, an average effect size includes different implementation contexts, which in turn generate further variance.Therefore, a meta-analytical approach was chosen, where each P.A.R.T.Y. day was regarded as an intervention in its own right. That is, we estimated for each included P.A.R.T.Y. day its own effect size and calculated the average effect size across all 19 individual P.A.R.T.Y. days by means of a meta-analysis. This overall mean effect size represents the average of the true effects within the sample. This approach allows for handling the expected variability of effect sizes and the uncertainty of estimates  appropriately. It also allows for taking the different sample sizes and study designs of each P.A.R.T.Y. day into account.In a first step, we began by carrying out descriptive analyses: For each of the 19 P.A.R.T.Y. days, mean value differences and their standard deviations for intervention and control group were calculated between the baseline and post-intervention survey as well as between baseline and the follow-up survey. From the descriptive data, the effect size Hedges’ g was calculated for each measuring instrument and each P.A.R.T.Y. day according to Borenstein et al. . StatistIn further analyses, the Mplus computer programme was used for estimating path models. These models describe possible relations between the scales explaining the impact of P.A.R.T.Y. on pupils’ self-reported behaviour. It follows the potential impact mechanisms introduced in chapter 2. Therefore, four models were estimated and compared. To reduce the complexity of the models we reduced the four dependent behavioural measurements to two constructs: approved behaviour  and disapproved behaviour (violations and dangerous play). Independent path models for both study waves were estimated instead of conducting cross-legged panel analysis to reduce complexity. The models include dependent mean differences of many variables at once, including the influence of participation in P.A.R.T.Y. as a dummy variable.By transferring the effect sizes into forest plots the variation of the single effect sizes became apparent, confirming the choice of our analysis strategy. For example, in Fig. mean = −.61). Thus, the P.A.R.T.Y. intervention most strongly affects pupils’ perception of the severity of accident injuries. It is noticeable that P.A.R.T.Y. obviously has no effect on cognitive processes like behavioural attitude and self-efficacy respective behaviour control. Likewise, P.A.R.T.Y. seemed to affect the perception of which behaviours one’s peers are typically performing (descriptive norm), but not of which behaviours are approved or disapproved by these peers (injunctive norm).Table mean = −.37).However, comparing the T0 baseline data with the T2 follow-up data – collected four to 5 months after the P.A.R.T.Y. days – there is only one statistical mean difference remaining. Only the perceived severity of accident injuries still shows a small positive effect in the long term  and behavioural intentions. In the medium term, however, only one of the effects remains significant. Hence, the aim of the following path models was to identify possible explanations for the effects that occurred in the short term but did not last.The correlation matrices of the mean scale values at all three survey points are presented in the Supplementary Tables Based on the mean differences between the T0 and T1, respective T2 path models were estimated. Figure As Fig. It is noticeable that not in all cases the statistical intervention effects of P.A.R.T.Y. in the path models correspond to the meta-analytical results presented above. For example, the path model Fig.  shows anThe model for the T0-T2 period showed a similar picture Fig. , even wiIn summary, the results show that the intervention does not directly increase feelings of threat. However, P.A.R.T.Y. obviously affected a theoretical determinant of the threat, namely the perceived severity whose change is directly related to a change in approved behaviour. Nevertheless, in turn it does not increase a person’s confidence in his or her ability to counter this threat actively (self-efficacy). This already provides one possible explanation for why the P.A.R.T.Y. does not have the intended effects on behaviour.p < .001). The intention in turn affected approved behaviour, but not disapproved behaviour. Conversely, the direct effect of self-efficacy on behaviour assumed in theory can only be proven for disapproved behaviour. The theoretically assumed model does not fit the data well. In order to achieve a better model fit, the direct relation between subjective norm and disapproved behaviour was added to the model even though it is not included in the theory.Figure The data of the second survey period T0-T2 reflect the Theory of Planned Behaviour somewhat better Fig. . This moIn both models, the measured changes in the predictors from the Theory of Planned Behaviour can explain more than 20% of the variance of the change in intention.Immediately after the intervention, small positive effects could be shown for most of the parameters. These effects could not be observed in the medium term and especially not for self-reported behaviour.Both in the short and medium term, the results showed a significant effect only on the threat-related characteristic perceived severity of accidental injuries.In the path analyses, the predictive influence of cognitive beliefs in particularly on a change in behavioural intention could be confirmed, such as self-efficacy or social norms. However, the P.A.R.T.Y. programme had no or only short-term effects on these factors.The aim of the present study was to evaluate the effectiveness of the P.A.R.T.Y. programme in Germany. The main results can be summarized as follows:Despite the worldwide dissemination of the P.A.R.T.Y. programme, there are only few methodologically sound studies about its effectiveness and no study investigating the impact mechanism by which the behaviour change is supposed to be achieved. Even though the findings of the international evaluation studies are difficult to compare due to different methodology, this P.A.R.T.Y. evaluation study showed similar results with regard to behaviour-related outcomes. We found small effects for self-reported behaviour immediate after the P.A.R.T.Y. intervention. This is in line with studies whose study period was rather short and lasted at most up to 1 month after the intervention , 21, 22.In contrast to the lack of efficacy with regard to attitude in the present study, some studies report at least short-term effects , 18, 57.Whether international P.A.R.T.Y. program versions achieve a change in other behavioural determinants like social norms or self-efficacy remains unclear, as these factors has not been included in previous effectiveness studies.Our results show that it seems unlikely that the P.A.R.T.Y. intervention programme in its current form will achieve its goal of reducing the risk behaviour of young people in traffic. An important reason for this seems to be that the assumptions on which the programme is based are insufficient, namely that young people can be motivated to change risky behaviour by strong emotions such as fear. Fear appeals may arouse the interest and attention of young people as a kind of “door opener” but without strengthening psychosocial beliefs namely self-efficacy, they seem to be more a “flash in the pan”. Thus, the PARTY program seems to be classified in a series of educational measures to change behaviour, which produce short-term effects, but for which no behaviour-related effects can be proven in the long term , 12, 13.Following the debate on fear appeals mentioned at the beginning of this paper, the models of the threat components nevertheless showed that perceived severity is able to influence self-efficacy, behavioural intention and even behaviour. Thus small, short-term effects can be achieved by communicating fear appeals. But the conditions for this should be further researched. Furthermore, according to Witte , in combThe results indicate that there needs to be elements in the programme to strengthen the cognitive beliefs in young people of avoiding risk behaviour in traffic. According to Bandura , a lastiThe social norms approach e.g. 59) could be used to influence descriptive norms regarding risk behaviour in the peer group. The approach is based on scientific evidence that suggests that students often overestimate the substance consumption of their peers. This overestimation can lead to an increase in their own consumption  could be. This deWe used a meta-analysis based on a random effects model to control the sources of variance within and between the classes or clinics that participated in the study. However, this method does not control bias at the beginning of data collection, namely in the selection of study units. The data are only based on a quasi-experiment, which means the school classes have been chosen by the schools for both intervention and control group. Preferably, the classes in all schools would have been assigned randomly to either the control or the intervention group . As this was only possible in one school, the equivalence of control and treatment conditions before the intervention could not be fully ensured as indicated by statistically significant age differences between school classes before the intervention.The reliability of some of the measurement scales is rather low, especially the reliability of the self-efficacy scale. Since the reliability of measuring instruments depends not least on the number of items used to measure a construct, there is often a conflict between reliability (i.e. number of item) and feasibility  of the questionnaire in intervention studies. The questionnaire could not exceed a certain length to be acceptable for students and schools. Therefore, we had to limit the number of items used to measure each construct. Because of the lack of an explicit programme theory, we needed to include a broad set of constructs from different theoretical models. That left only few items per construct. For self-efficacy, for example, there are various dimensions discussed in the literature . The lowSummarising the results from this study the following main conclusions can be drawn. Apparently, communicating threat alone is not sufficient to achieve a lasting change in self-reported traffic-related risk behaviour. In addition, it needs elements to change cognitive beliefs as well. In the short term, such elements should be developed in a theory-based manner. Most promising for behaviour change are elements enhancing self-efficacy or eliciting descriptive social norm. Cognitive processing could be facilitated with elements carried before and after the actual P.A.R.T.Y. day at schools, e.g. preparation material, refresher courses. An explicit programme theory should be developed and tested that could guide the revision of the programme. Meta-analytical procedures have proven to be appropriate for programme evaluations where differences across implementations are typical for the programme rather than an indication of inadequate implementation. There are randomized study designs and statistical procedures available not only on an individual level but for clusters (e.g. school classes). They allow for less biased effects, which gives a better picture of the effects and underlying impact mechanism of programme interventions.Additional file 1: Supplementary Table 1. Correlation matrix of the mean scale values to the baseline survey (T0).Additional file 2: Supplementary Table 2. Correlation matrix of the mean scale values to the post survey (T1).Additional file 3: Supplementary Table 3. Correlation matrix of the mean scale values to the follow-up (T2).


Pareuchiloglanis sichuanensis is an endemic fish species in the upper reaches of the Yangtze River. In the present study, the complete mitochondrial genome of P. sichuanensis was analyzed. The mitochondrial genome, consisting of 16,774 base pairs (bp), included 13 protein-coding genes, 2 ribosomal RNAs, 22 transfer RNAs, and a non-coding control region. The phylogenetic tree showed that P. sichuanensis was closely related to P. anteanalis. These results provid the useful information for further studies on taxonomic status, molecular systematics, and stock evaluation. Pareuchiloglanis sichuanensis Ding, Fu, Ye P. sichuanensis. In this study, we sequenced, assembled, and annotated the complete mitochondrial genome of P. sichuanensis, which could provide the useful genomic resources for the future studies.P. sichuanensis specimens were collected from the upstream of Tianquan River, a tributary of the Qingyi River  and chosen for mitochondrial genome analysis in 2020. The voucher samples were deposited at Aquaculture Department of Sichuan Agricultural University . Then, the total genomic DNA was extracted from the muscle by a traditional phenol-chloroform method. Whole genome sequencing was conducted with Illumina Novaseq platform. The mitochondrial genome was assembled de novo using A5-miseq(v20150522)  were encoded on the light-strand. On the contrary, all the other genes were encoded on the heavy-strand. This was a typical gene arrangement conforming to the other Pareuchiloglanis species and vertebrate consensus  under the accession no. MW697900.The complete mitogenome of Pareuchiloglanis, we selected seven Pareuchiloglanis species (Pareuchiloglanis species have the close relationship, and P. sichuanensis and P. anteanalis are monophyletic in the tree. Thus, the mitochondrial genome data and phylogenetic analysis of the P. sichuanensis enrich the evolution research of Pareuchiloglanis.To better understand the phylogenetic relationships of mitochondrial sequences in  species . Based o species  (Figure  species . The res


Seed shape in species of the Cactaceae is described by comparison with geometric models. Three new groups of models are presented, two for symmetric seeds, and a third group for asymmetric seeds. The first two groups correspond, respectively, to superellipses and the combined equations of two semi-ellipses. The third group contains models derived from the representation of polar equations of Archimedean spirals that define the shape of asymmetric seeds in genera of different subfamilies. Some of the new models are geometric curves, while others are composed with a part resulting from the average silhouettes of seeds. The application of models to seed shape quantification permits the analysis of variation in seed populations, as well as the comparison of shape between species. The embryos of the Cactaceae are of the peripheral type, strongly curved and in contact with the inner surface of the seed coat. A relationship is found between seed elongation and the models, in which the genera with elongated seeds are represented by models with longer trajectories of the spiral. The analysis of seed shape opens new opportunities for taxonomy and allows quantification of seed shape in species of the Cactaceae. The Cactaceae Juss., with about 2000 species distributed in ca. 120 genera, are characterized by succulent photosynthetic stems. The family has been included in the order Caryophyllales Juss. ex Bercht. & J.Presl ,4,5. Rhipsalis baccifera (J.S. Muell.) Stearn, found also in local populations in tropical Africa and Sri Lanka.Cacti are common in the arid ecosystems of America, in the southern States of USA and Mexico, north-eastern Brazil, and along the slopes of the Andes from Ecuador to Argentina and Chile. Many species also live in humid forests as epiphytes. All known species are originally American with the only exception of Maihuenia (Phil.) Britton & Rose, with two cushion-forming, mucilaginous species, is the only genus in the Maihuenioideae. Pereskia Mill, together with Leuenbergeria Lodé in the Pereskioideae, are woody with large, glossy leaves and long thorns on the stem, and not all are succulents. The Cactoideae, with ten tribes, is the most diverse subfamily of globular and columnar cacti including all the iconic species in American dessert biomes [Pterocactus, which possess flattened winged seeds [The Cactaceae have been divided into four subfamilies: Cactoideae Eaton, Maihuenioideae P. Fearn, Opuntioideae Burnett, and Pereskioideae Engelm. The Cactoideae and Opuntioideae contain, respectively, 100 and 9 genera ,5. Maihued seeds .The flowers of the Cactaceae have interesting peculiarities, such as the presence of the perianth and the pericarpel, the stem-tissue enclosing the inferior ovary ,7.Cereus [Blossfeldia and the Opuntioideae [Difficulties for taxonomy in this family are partly due to three reasons: (1) A large amount of homoplasy in their characters ; (2) TheCereus ,11,12, atioideae ,14.Ferocactus Britton & Rose [Pachycereus (A. Berger) Britton & Rose [Neobuxbaumia Backeb. [Stenocereus (A. Berger) Riccob. [Melocactus Link & Otto [The analysis of seed surface structure by electron microscopy has given useful information in the taxonomy of n & Rose , Pachycen & Rose , Neobuxb Backeb. , Stenoce Riccob. , and mork & Otto . NeverthBlossfeldia liliputana); 2: lenticular (Pereskia bleo); 3: pyriform (Escobaria cubensis); 4: mussel-shaped (Selenicereus grandiflorus); 5: reniform (Neobuxbaumia multiareolata); 6: hat-shaped (Frailea phaeodisca). This classification is similar to others, see for example [According to seed shape, Barrios et al.  classifi example ,21, shar example ,23. Morp example ,35,36,37Arabidopsis thaliana (L.) Heynh., the model legumes Lotus japonicus L. and Medicago truncatula Gaertn., as well as Capparis spinosa L., in the Capparaceae and Rhus tripartita (Ucria) Grande in the Anacardiaceae [Triticum sp. in the Poaceae and Ricinus communis L. and Jatropha curcas L. in the Euphorbiaceae were described by the similarity of their images to ellipses of varied x/y ratio [Models based on the cardioid or modified cardioids were first applied to the description of seeds of the model plant rdiaceae ,26,27,28/y ratio ,30,31. A/y ratio . The Cuc/y ratio .A series of geometric models have been described also for the Arecaceae and the Vitaceae ,35. In tOur objective was to obtain geometric models for the description of seed shape in the Cactaceae. With this basis, seed shape description and quantification could be applied in taxonomy.Rebutia heliosa Rausch, R. pseudodeminuta Backeb., and R. fiebrigii (Gürke) Britton & Rose are shown in p = 2.3, a = 1.5, b = 2 for R. heliosa, p = 2.6, a = 1, b = 2.1 for R. pseudodeminuta, and p = 2.2, a = 1.5, b = 3 for R. fiebrigii.Seeds of Rebutia are symmetric. The seed images of many other species are also symmetric, but in many cases, they present a certain degree of differentiation with a section straight corresponding to the hilum cup, and a rounded part in the opposite pole. This type of seeds is associated with a short funicle and were observed in species of Echinopsis, Melocactus, and Thelocactus.The models for the seeds of three species of Echinopsis, three models were identified for E. calochlora, E. leucantha, and E. klingeriana, obtained, respectively, from curves described by the following equations:For the seeds of The three models are represented in Figures resulting from the representations of the Archimedean (in wide sense) spiral ∈0.1, 20 A, t∈0.1,t∈0.1, 5 B, t∈0.3,t∈0.3, 5 C, t∈0.5,t∈0.5, 5 D, and t∈ , perimeter (P), length (L), width (W), aspect ratio (L/W), circularity, and roundness in the seeds are shown in F. herrerae, E. platyacanthus, and P. pringley formed a group with smaller area values. The seeds of Pereskia bleo stand out as having higher area values, according to their reduced number of seeds per fruit (see later). The seeds of H. undatus and P. bleo had lower circularity. In respect to roundness, there were no statistical differences between O. ficus-indica and E. platyacanthus nor between P. pringley and F. herrerae. The differences between the values of circularity and roundness were due to irregularities in the seed surface, which increased the perimeter values, decreasing roundness. From the obtained data, it can be concluded that different models should be applied to each species. There were differences in size and shape between seeds of different species. The seeds of F. herrerae and 12.84 for the same magnitude in P. bleo  was 91.2 . The seeds of P. pringlei adjusted well to the model. The coefficient of variation for the J index was 2.19.F. herrerae. Mean value of J index in an observed group of 20 seeds (with the corresponding model) was 91.2 . The seeds of F. herrerae adjusted well to the model. The coefficient of variation for the J index was 3.73.E. platyacanthus than in the other species analyzed so far . This is due to the disposition of seeds in the fruit, being densely packaged with different orientations .Visual inspection of the seeds revealed more heterogeneity for shape in the seeds of ntations A. FigureO. ficus-indica. Mean value of J index in 20 seeds  was 91.2 . The seeds of O. ficus-indica adjusted well to the model. The coefficient of variation for the J index was 2.52.Geometric models derived from the Archimedean spiral defined well the silhouette of the seeds of Maihuenia and Pereskia  shares a similarity with the general model described for Opuntia species in the Opuntioideae, based on fragments of an Archimedean spiral. Seed shape may share features between these three subfamilies with many plesiomorphic characters, such as the presence of leaves and shrub habit [To explore seed shape in the Cactaceae, we have considered plant material belonging to the main subfamilies, with emphasis on the Cactoideae because it comprises about 80% of the total diversity . A preliub habit . This geRebutia (Trichocereae), Hylocereus undatus (Hylocereae), Pachycereus pringlei (Pachycereae), Ferocactus herrerae, and Echinocactus platyacanthus (Cacteae). The observation of seeds belonging to other species in these tribes suggests that there may be some correlation between seed shape of different species from the same tribe. For example, the seeds of Disocactus nelsonii (Britton & Rose) Linding, Selenicereus grandiflorus (L.) Britton & Rose, and S. inermis Britton & Rose (Hylocereae) have an overall shape similar to Hylocerus undatus. In addition, the species of the two genera belonging to the same tribe (Ferocactus herrerae and Echinocactus platyacanthus) had the same geometric figure as the basis for the respective models, and other and genera such as Mammillaria Haw. in the Cacteae share a similar form. The equations presented may be the basis for the design of models specific for tribes or genera, but this task requires the analysis of a number of seeds.The Cactoideae is the subfamily with the highest number of life forms, morphological types, and species richness ,39, and Rebutia (Aylostera). A second group is represented by seeds whose shapes are also symmetric, but non-convex. These types of seeds are characteristic of some species in the Cactaceae and present a straight part in the hilum cup, and a rounded dorsal part, such as in species of Echinopsis, Melocactus, and Thelocactus, corresponding to the space occupied by the peripheral embryo [The seeds of the Cactaceae present an interesting diversity in shape. Forms that are frequent in other plant families, such as ovals and ellipses ,40, are l embryo ,42.Coriaria L., such as C. arborea Linds. and C. japonica A. Gray , as well as in Sagittaria longiloba Engelm. ex J.G. Sm. and Echinodorus tenellus (Mart. ex Schult.f.) Buchenau  [Opuntia species, a turn is visible in O. ficus-indica seeds, where the funicle surrounds the ovule completely being like a thick third integument [Seeds in other species of the Cactaceae present characteristic shapes resembling the generalized Archimedean spirals with equation =tα, see . Seeds wmatales) . We are tegument . It is itegument . Chaptertegument  is dedicThe reniform shape, better defined as the cardioid curve, is also discussed by D’Arcy Thompson in chapter 9 of his book . This chArabidopsis. The formation of phyllotactic spirals in Arabidopsis, similar to the development of the inflorescences in the cauliflower, have revealed a complex set of gene interactions [Pterocactus [The molecular basis of the formation of developmental patterns acquired a great impulse with the application of genomics and the analysis of mutants in ractions . The sharocactus  are key Echinocactus platyacanthus Link & Otto and Ferocactus herrerae J.G.Ortega were obtained from plants grown in the campus of University of Alicante ; the seeds of Pachycereus pringlei (S.Watson) Britton & Rose were from a fruit grown in El Huerto del Cura ; and the seeds of Hylocereus undatus (Haworth) D.R. Hunt were obtained from a commercial pitahaya bought on a local market in Albir . All the fruits mentioned above were collected between 6 August and 12 August 2021, and their seeds photographed in September, 2021. Opuntia ficus-indica (L.) Mill seeds are from plants grown in Ambato, Tungurahua (Ecuador) and Villar de Peralonso, Salamanca (Spain) collected in 2020. Pereskia bleo (Kunth) DC seeds were collected in a public garden close to the Malecón Boayaku at Puyo  on 12 October 2021 and photographed in the next days. After being photographed, the seeds were returned to their origin. A sample of this plant will be deposited in the Amazon State University Herbarium (ECUAMZ).To explore seed shape, plant species belonging to the main subfamilies, with emphasis on the most diverse Cactoideae, have been considered. The seeds of Rebutia K. Schum., Echinopsis Zucc., Maihuenia (Phil. ex F.A.C.Weber) K. Schum., and Thelocactus Britton & Rose were obtained from https://cactus-aventures.com/TaxonomySeedGallery/Seeds%20album/ (accessed on 27 September 2021). On this webpage, Rebutia species are shown with the name Aylostera, which is a synonym. The images of seeds of O. ficus indica and P. bleo, which were collected in Ecuador, the photographs were taken with a camera Cannon PowerShot SX520 HS.Photographs were taken with a camera Z6 with an objective AF-S Micro NIKKOR 60 mm f/2.8G ED. Composed images containing 10–30 seeds per accession were prepared with Corel Photo Paint. For Values of the area (A), perimeter (P), length (L), width (W), aspect ratio , circularity (C), and roundness (R) of the seed images were obtained in Image J .https://zenodo.org/record/4478344#.YBPOguhKiM8, accessed on 9 November 2021). The layers containing the seeds were superimposed and the opacity is given a value of 30 in all layers. All the layers were combined, and the brightness is adjusted to a minimum value. From this image, we are interested in the inner region representing the area where most of the seeds coincide, which is the darkest area. To select it, we use the magic wand tool, and with a tolerance equal to 10, this selection is copied and pasted as a new layer. Twenty seeds were used to obtain the average silhouettes for Echinocactus platyacanthus, Ferocactus herrerae, Hylocereus undatus, Opuntia ficus-indica, and Pachycereus pringley. For Echinocactus platyacanthus, the average silhouettes were obtained separately from 20 seeds of long and short seeds. For Pereskia, average silhouettes were obtained from 9 seeds.The average silhouette is a representative image of seed shape for each group of seeds. It was obtained in Corel Photo Paint, by the protocol described in   and R. software v. 4.0.5 , perimeter (P), length (L), width (W), aspect ratio , circularity (C), and roundness (R) as well as the eam 2020 . Accordiit × 100 .Echinocactus, Ferocactus, Hylocereus, Pachycereus, and Pereskia. The seeds of Opuntia ficus-indica adjust well to a model derived from equation Rebutia species were based on the equations for superellipses, while for Echinopsis, the models derived from the representation of two semi-ellipses. The comparison with geometric models is proposed as a basis for seed description and quantification in the Cactaceae.New models based on geometric figures are proposed for seed shape description and quantification in species of the Cactaceae. Models derived from equation


Anopheles species: An. atroparvus, An. hyrcanus sensu lato (s.l.), An. labranchiae, An. maculipennis sensu stricto (s.s.), An. messeae/daciae, An. sacharovi, An. superpictus and An. plumbeus. The distribution of these potential vector species across Europe is critically reviewed in relation to areas of major presence and principal bionomic features, including vector competence to Plasmodium. Additional information, such as geographical details, sampling approaches and species identification methods, are also reported. We compare the information on each species extracted from the most recent studies to comparable information reported from studies published in the early 2000s, with particular reference to the role of each species in malaria transmission before eradication. The picture that emerges from this review is that potential vector species are still widespread in Europe, with the largest diversity in the Mediterranean area, Italy in particular. Despite information on their vectorial capacity being fragmentary, the information retrieved suggests a re-definition of the relative importance of potential vector species, indicating An. hyrcanus s.l., An. labranchiae, An. plumbeus and An. sacharovi as potential vectors of higher importance, while An. messeae/daciae and An. maculipennis s.s. can be considered to be moderately important species. In contrast, An. atroparvus and An. superpictus should be considered as vectors of lower importance, particularly in relation to their low anthropophily. The presence of gaps in current knowledge of vectorial systems in Europe becomes evident in this review, not only in terms of vector competence but also in the definition of sampling approaches, highlighting the need for further research to adopt the appropriate surveillance system for each species.Despite the eradication of malaria across most European countries in the 1960s and 1970s, the anopheline vectors are still present. Most of the malaria cases that have been reported in Europe up to the present time have been infections acquired in endemic areas by travelers. However, the possibility of acquiring malaria by locally infected mosquitoes has been poorly investigated in Europe, despite autochthonous malaria cases having been occasionally reported in several European countries. Here we present an update on the occurrence of potential malaria vector species in Europe. Adopting a systematic review approach, we selected 288 papers published between 2000 and 2021 for inclusion in the review based on retrieval of accurate information on the following The online version contains supplementary material available at 10.1186/s13071-022-05204-y. Plasmodium parasites, remains a major healthcare challenge in tropical regions, particularly in sub-Saharan Africa, where Plasmodium falciparum and Plasmodium vivax are responsible for most of the morbidity.Despite major efforts of researchers and funding institutions to tackle malaria, the most deadly of known parasitic diseases, the global burden of this life-threatening disease remains dramatic. However, great strides in combatting malaria were made in the last century, with the disease eradicated from several regions of the world, in particular in temperate areas, thanks to joint efforts at scientific, social and political levels. This disease, caused by five different species of Despite the eradication of malaria across most European countries in the 1960s and 1970s, the anopheline vectors of this disease are still present in Europe .Anopheles species competent for malaria transmission, most contain detailed information on countries where malaria is endemic ) in this review based on the findings of recent studies indicating their potential role as malaria vectors in some geographic areas. Lastly, Anopheles maculipennis sensu stricto (s.s.) is also taken into consideration; although this species is considered to be a secondary malaria vector, its wide distribution and local abundance make it a potential malaria vector in some areas. The criteria of selection of species are based on their occurrence, abundance, biology, evidence or suspicion of involvement in malaria transmission  and vectorial competence. To retrieve a more complete set of data, the taxonomic definition An. maculipennis s.l. was also searched. Some authors formally state that An. daciae is a new species  this result is of great importance in explaining the presence of this species in the area even when rice paddies are dry (end of summer and autumn)”. In agreement with this evidence, An. hyrcanus s.l. larvae reported in countries of central and southeastern Europe were found mainly in large water collections, such as pools, fishponds and swamps [High densities of  species , 77, 78.d swamps , 79–82.Anopheles hyrcanus s.l. adults are exophilic, with outdoor resting sites during the day. Gadzhieva [An. hyrcanus s.l. was influenced by environmental humidity; consequently, it is a species for which the proximity of the breeding sites is linked to the hygrophilous vegetation.adzhieva  made detDiapausing females were observed to mainly exploit natural shelters, including clefts in the ground, grasses and reed bushes, while indoor-resting specimens were rarely observed .Anopheles hyrcanus s.l. has historically been described as strictly mammalophilic, with a high degree of anthropophily. This behavior is confirmed by several recent studies that performed human landing catches and used animal baited traps [Anopheles hyrcanus presented a huge anthropophily with spectacular aggressiveness on humans: during this study, scientists underwent massive attacks from females of this species”. Moreover, Aldemir et al. [ed traps , 70, 71.ed traps  reportedr et al. , 70  [P. vivax seems possible. In fact, An. labranchiae has been implicated in autochthonous cases of malaria by P. vivax that have occurred both in Corsica and Italy [In the past, in NF54) , 89. On nd Italy –97, indiAnopheles maculipennis s.s. is probably the most widespread Anopheles species in Europe .Anopheles maculipennis s.s. is known to be endophilic, resting in stables and dwellings. In recent literature, this species was reported to have been collected in large numbers in animal shelters, in particular those with pigs or sheep present [ present , 110. DiAnopheles maculipennis s.s. is considered to be strongly zoophilic and, therefore, to play a secondary role in malaria transmission. However, in specific contexts , the species has been found to show a certain degree of anthropophily [An. maculipennis s.s., although considered mainly zoophilic, also results as being very aggressive on humans during the night catches, both in presence or absence of animals” [opophily . Recent opophily , 111  assessed its occurrence in a number of countries, resulting in an increased number of studies focusing on An. messeae or An. daciae. However, the resulting literature was mainly based on the description of the molecular identification of An. daciae, with little additional information, and usually limited to reporting the location where the specimens were collected. Few articles reported ecological and biological differentiation between An. messeae and An. daciae, such as reproductive isolation, feeding behavior, vector competence, morphology, etc.Following the definition of Anopheles messeae/daciae is one of the most widespread species of the Anopheles maculipennis complex in Europe  waters, while An. daciae was found in atypical breeding sites, in waters with low oxygen content (0.8 mg/l).Few articles have reported distinct data for ng sites , 121. Bopastures , 121. Napastures  describeAnopheles messeae/daciae have been found resting outdoors and to be particularly abundant in animal stocks, which is in agreement with the strong relationship between the high abundance of this species in rural/natural habitats and the presence of animal shelters in these areas [Adults of se areas , 125. Fese areas , indicatAn. messeae/daciae diapause was not retrieved. Based on previous descriptions, during the winter this species enters into full hibernation at the adult stage without any feeding activity, resting in enclosed shelters or buildings [New information on uildings .An. messeae/daciae does not feed on humans and, therefore, considered it to be a malaria vector of negligible importance [Anopheles messeae/daciae was captured in animal housing that sheltered different animal hosts, such as cows, horses, sheep and goats, but the results of this study did not unequivocally demonstrate whether this was a consequence of sampling bias or host choice [An. messeae/daciae blood meals indicated cattle and dog as major hosts [This species was described in the past as a strongly zoophilic feeder. The authors of one article stated that portance . Severalt choice . Similart choice , 125. Inor hosts , 127. Itor hosts , and thior hosts .An. messeae and An. daciae to be separate species, limited and inconclusive evidence can be obtained in the retrieved literature on their respective feeding pattern. According to Danabalan et al. [, An. daciae may feed on animals and humans, whereas An. messeae s.s. appears to be strictly zoophilic. However, these observations were based on a single study that collected only a few samples and performed in a small area. Other authors speculated that An. daciae is more anthropophilic than An. messeae because it was found more frequently in larval habitats located close to human dwellings [An. daciae blood-fed only on wild mammals, with no human blood detected, while An. messeae blood-fed on both birds and mammals, humans included [Considering n et al. , An. dacwellings . On the included .An. messeae/daciae as a malaria vector remains controversial. It seems refractory to tropical P. falciparum strains , but itn Europe , 64. In n Europe  attempte messeae , 121. Coe Tables .Anopheles species recorded in Europe, An. sacharovi has the most southern distribution , with only one female having had a blood meal on a human. Similarly, Tavşanoğlu and Çağlar [An. sacharovi fed preferably on animals rather than humans even if mosquitoes were collected in houses. However, the results of these studies were affected by the proximity of the houses and animal sheds, both usually being open during the summer, thereby allowing free movement of blood-fed females searching for a suitable resting place  in those areas [Plasmodium never being detected in field mosquitoes. However, this species was demonstrated to be experimentally competent not only for P. vivax [P. falciparum [The role of acharovi . In suppse areas , 136, deP. vivax  but alsolciparum , indicatAn. superpictus be considered a species complex [An. superpictus described in older publications refers to studies conducted outside of Europe [An. superpictus findings in the recent literature came from southeastern Europe, except for one study in Italy [There are suggestions in the extracted literature that  complex . Its widf Europe . All An.in Italy  where a An. superpictus are temporary pools formed by fluctuations in the water level of rivers or streams caused by rain activity. Such sites are typically described as pools of clean water with gravelly riverbeds exposed to sunlight [An. superpictus in man-made habitats, such as irrigation channels, rice fields, ditches and artificial pools [An. superpictus in breeding sites with low salinity and a chloride concentration ranging between 30 and 96 mg/l [Typically, the breeding sites of sunlight . Severalal pools . Other lal pools , 140, weal pools  collecte 96 mg/l   where there was an important malaria focus” [The role of P. vivax  and the P. vivax . Only ona focus” ; howeverAnopheles plumbeus is one of the most diffused species among those targeted in this review. It is widespread throughout Europe up to the Caucasus and Asia, from colder to warmer climates  and are still present in European countries. In some areas of Europe they are also abundant in terms of species diversity, particularly in southern Europe .An. sacharovi). This reduction is mostly due to human interventions that modified the original larval habitats , and to the increasing usage of insecticides. Several Anopheles species  now exploit alternative larval habitats linked to some agricultural practices (e.g. rice cultivation and the use of extensive irrigation networks) that have increased in recent years, thereby allowing for the presence of mosquito populations also in anthropized areas. In particular, anthropic habitats are now exploited by new potential malaria vectors, as shown by the presence of An. plumbeus, which is now able to exploit urban environments using small water containers rich in organic matter as habitats. This indicates that some potential malaria vectors possess a non-negligible ecological plasticity.Despite this abundance, at the ecological level a reduction in their original habitats has been observed, as compared to the past , sampling technique(s) and geographical details of sampling sites. When the presence of Anopheles species was not assigned to a specific site, all the sites monitored during that study were considered to be negative and marked as NA. Abbreviation: DD, Decimal degree; NA, not available.


Pteris vittata in the As–Pb–Hg-polluted Bone River area, Gorontalo Province, Indonesia. The density distribution of P. vittata decreases from around the artisanal and small-scale gold mining (ASGM) site to the lower reaches of the Bone River, and it is rarely found near Gorontalo City. The maximum concentrations of As, Hg, and Pb recorded in the soil samples were 401, 36, and 159 mg kg−1, respectively, with their maximum concentrations in P. vittata recorded as 17,700, 5.2, and 39 mg kg−1, respectively. Around the ASGM sites, the concentrations of As, Pb, and Hg in P. vittata were highest in the study area. These data suggest that P. vittata, a hyperaccumulator of As, may be useful as a bioindicator for assessing environmental pollution by Pb and Hg.In this paper, we report ecological and environmental investigations on  Artisanal and small-scale gold mining (ASGM) is one of the largest emitters of mercury (Hg) into the environment in the world. In ASGM, miners use elemental Hg to extract gold (Au) from finely crushed rocks or Holocene sediments, forming an Hg–Au amalgam. This amalgam is then burned, releasing Hg as vapor and leaving impure Au residue. The adverse effects on the health of miners are severe, with exposure to Hg causing neurological damage and other health issues. The people residing near the ASGM sites are also affected by the Hg contamination of water, air, and soil and by the effects of Hg on ecosystems, such as fishes and plants. Approximately 15 million people in ASGM sites in over 70 countries use Hg to refine Au, causing Hg pollution [In Indonesia, over 300,000 ASGM miners work at approximately 1000 informal sites. These miners are either displaced farmers or fishermen who have very few options in terms of alternative livelihoods. These rural communities depend on the ASGM for income. The practice of using Hg to extract Au was banned by the Indonesian government in 2014 [According to the data of the Ministry of Forestry and Mining of Gorontalo Province in 2012, Gorontalo Province has a large number of ASGM sites, especially in the Bone Bolango Regency, where the largest number of ASGM workers work . AccordiIn recent years, although many researchers have focused on Hg in environmental assessment studies at ASGM sites, little attention has been given to the other toxic metals associated with Au deposit formation. In particular, Neogene epithermal gold deposits in the Circum-Pacific Rim  are knowPteris vittata is a fern species with a hyperaccumulation ability for As, and it can accumulate up to approximately 28 g/kg As in its fronds [P. vittata has been recently developed as an environmentally friendly and cost-effective technology to remediate heavy metal-contaminated environments [Generally, higher plants find it extremely difficult to survive in environments that are highly contaminated with As. However, s fronds ,12. It cronments .Pteris. vittata is known to be an accumulator of metals in laboratory experiments. For example, Fayiga et al. demonstrated the capability of P. vittata to hyperaccumulate arsenic from soils in the presence of Cd, Ni, Pb, and Zn in a greenhouse study [P. vittata acts as an “indicator plant” in severely contaminated soils in ASGM and examines the potential to remove Pb, As, and Hg.se study . The novP. vittata to remediate contaminated soils in China has also been empirically examined [P. vittata in As-contaminated soils through a 2-year field experiment [P. vittata as an accumulator of metals, limited studies have examined the role of P. vittata as an accumulator of metals in the case of ASGM locations. Soils of ASGM locations are usually polluted by historical mining operations, and the potential of bioaccumulators of P. vittata are influenced by the level of pollutants and climatic and geographical conditions. Further empirical studies are necessary in ASGM locations to examine the potential of accumulating metals from contaminated soils. Therefore, the present study yields important policy implications for achieving sustainable livelihoods through adopting environmentally friendly technologies.The potential of examined ,16,17. Fperiment . Since pP. vittata and analyzed the metal concentrations of the samples of soils and P. vittata in ASGM areas polluted by multiple metals, such as Hg, As, Zn, and Pb, around the Bone River, Gorontalo Province, Indonesia. We discussed the correlations between the distributions of P. vittata and the metal concentrations between the soils and plants.In this study, we conducted a field survey of the distribution of the population density of Over the past decade, mine tailings and wastewater from the two ASGM sites located upstream of the Bone River and smelted slag from the sites along the valley have been dumped along the river, and they are mixed into the river water.2, and altitudes can reach up to 1500 m. The two ASGM sites in this region are Motomboto and Mohutango. P. vittata and soil samples were collected along the Bone River from downstream to upstream close to the ASGM areas . The regency has an area of 1985 kmGM areas .P. vittata along the Bone River from the ASGM area to Gorontalo City. Plant population density was determined using the quadrat counts method [We mapped the distribution of the population density of s method  by countn = 20) were collected from depths of 10–20 cm from the P. vittata collection points. The soil samples were dried (120 °C for 48 h) with a ventilated oven ISUZU drying oven Model 2-2045 and stored in sterilized plastic bags prior to analysis. The dried soil samples were sieved using a 2 mm sieve for the removal of larger organic components, and the smaller components were removed using tweezers. The samples were ground to a powder in a planetary ball mill  and homogenized before analysis by particle-induced X-ray emission (PIXE) at the Cyclotron Research Center, Iwate Medical University, Japan. All the samples were imported into Japan under a permit issued by the Ministry of Agriculture, Forestry, and Fisheries, Japan, in accordance with the Plant Protection Law.In all, 37 plant samples were collected from the field from upstream to downstream along the Bone River at appropriate intervals, and one whole plant was taken at each point. The plant tissue we used for analysis was the fronds. Soil samples  performed using a Varian 820-MS instrument  at the Integrated Center for Sciences, Ehime University, Japan. The flowchart of the methodology is illustrated in The plant samples were washed with deionized water and dried in a ventilated oven at 80 °C for 48 h. The dried leaves were ground to a powder in a ball mill and prepared for analysis as follows: the sample powder (20 mg) was digested with 1 mL HNOThe US National Institute of Standard and Technology (NIST) CRM 1643f  and multThe significant differences among the sampling sites were determined using Stata 16.1  and OriginLab v. 9.60 program for Windows.P. vittata in the study area  (1993)  are listEPA 1993 . The oveP. vittata and soil samples are summarized in −1, which was relatively low but tended to increase toward the ASGM sites.The concentrations of As, Pb, and Hg in the samples of P. vittata were in the ranges of ND–17,700, ND–39 mg kg−1, and ND–5.2, respectively. The ND data correspond to a situation in which the concentration is lower than the detection limit of the analytical instrument.The concentrations of As, Pb, and Hg in P. vittata and the As concentration in soil. This correlation is also consistent with P. vittata being found in As-contaminated areas [P. vittata grows actively in a neutral or slightly alkaline environment [There is generally a positive correlation between the density distribution of ed areas . P. vittironment ,25. AlthP. vittata absorbed large amounts of As, reaching a maximum concentration of 17,700 mg kg−1 (We found that  mg kg−1 . The rocP. vittata and As concentration in the rhizosphere soil (P. vittata indicate that the removal efficiency of P. vittata depends on the ratio of soluble As and the pH range of the soil [The results of this study show dispersion and no significant correlation between ere soil . The resthe soil ,13,25. TP. vittata in the mining areas was higher than that in the downstream area. The source of the high concentration of Hg in the ASGM area was mainly Au extraction activities. Hg could be transferred in forms such as methylmercury in the ecosystem [P. vittata. Based on the ability to absorb Hg, P. vittata is considered to have the potential for use as a bioindicator for Hg [Generally, Hg accumulation in the soil is related to deposition caused by anthropogenic activities through the biosphere or atmosphere . The ASGcosystem  and was r for Hg ,32.P. vittata had a maximum Pb concentration of 39 mg kg−1 that tended to be considerably higher around the ASGM sites. These results indicate the possibility of using this plant as a bioindicator for Pb in contaminated areas.The maximum soil concentrations of Pb in this study area are higher than the limit prescribed by the US EPA for agricultural soil . P. vittP. vittata are shown in The correlations among As, Hg, and Pb concentrations in P. vittata and soil samples is shown in The correlation between As, Hg, and Pb in P. vittata can adopt and survive in metal-contaminated soils, there is a concern that it could be an invasive species, which thereby influences local biodiversity. Rathinasabapathi discusses the competitive ability and invasiveness of several ferns [P. vittata in the surveyed areas, further studies are required to understand the influences of expanding P. vittata for other local species.The findings raise concern for biodiversity in the local environment. Since al ferns . As therP. vittata. Examining the composition of mobile fractions extracted from environmental solid samples may provide a clearer explanation of the uptake. There are two reasons why the study did not examine the composition of mobile/bioavailable fractions. First, the current study provides a relationship between uptake and total metal contamination, even though mobile/bioavailable composition is not fully investigated. More total metal contamination is identified, and more plant uptake is empirically identified. Second, even mobile/bioavailable and solid fractions are mixed in the field, providing evidence for total metal contamination and plant uptake yields important implications for policy-makers, especially in developing countries, regarding whether contamination is prevalent. At this point, the research is considered a preliminary investigation, and further research is necessary. Examining the composition of mobile fractions from environmental solid samples to show clear characteristics between metal compositions and plant uptake is beneficial to elaborate on the findings of the research.The study did not examine the mobile/bioavailable fraction of metals that could potentially correlate with uptake by P. vittata in the As–Pb–Hg-polluted Bone River area, Gorontalo Province, Indonesia. The density distribution of P. vittata decreases from around the ASGM sites to the lower reaches of the Bone River, and the plant is rarely found near Gorontalo City. The maximum concentrations of As, Pb, and Hg in P. vittata were 17,700, 5.2, and 39 mg kg−1, respectively. Around the ASGM sites, the metal concentrations in P. vittata were the highest in the study area. These data suggest that P. vittata, which is a hyperaccumulator of As, may also be useful as a bioindicator for assessing the environmental pollution caused by Pb and Hg.We conducted ecological and environmental investigations on P. vittata acts as an indicator plant in other areas. Identifying the potential and limitations of P. vittata as an indicator plant by extensive investigations will be helpful for policy-makers to assess the environmental pollution of respective ASGM areas.Since this research was limited to case studies in the ASGM regions of Gorontalo Province, Indonesia, further research is pivotal for generalizing the results. For example, a high level of mercury contamination has been reported in ASGM communities in various provinces, such as in West Java , West Nu


Superficial mycoses are the fourth most common cause of disease worldwide. It is not surprising that zoonotic transmission occurs to humans due to close contact with different animals,be it companion or farm animals. Therefore, the objective of this study was to determine the presence of asymptomatic dermatophyte carriers in the owner-pet pairs, identify the most commonetiologic agents, and find the likely connection between the carrier status of an owner and the presence of dermatophytes in their pets.Trichophyton tonsurans and Trichophyton verrucosum, while in pets, a strain of Trichophyton sp was found. From May 2019 to January 2020, 21 cats and 115 dogs with their respective owners were selected for dermatophyte culture. All the dogs and cats included in the study were from thecommunities of southeastern Mexico. The samples were taken with a cotton swab, which was vigorously rubbed and twisted on the scalp or body of the pet four times and grown on Mycosel Agar.The isolates were identified based on macroscopic and microscopic characteristics. The prevalence of the binomial ranged from 0.73% in pet skin and human hands to 2.2% in human scalp.In humans, the agents were  Different species of dermatophytes were found in the owner/pet pairs, which denotes that coexistence is not related in asymptomatic cases. Dermatophytes are the most common cause of superficial fungal infections. They are a specialized group of filamentous fungi capable of infecting and degrading keratinized tissues,including skin, hair, and nail  . Every Microsporum canis from dogs and cats to their owners [ It is not surprising that zoonotic transmission occurs to humans due to close contact with either companion or farm animals. In this regard, Cafarchia et al.have described the transmission of  owners   and the owners  ; howeveTherefore, this study aimed to determine the presence of asymptomatic dermatophyte carriers in the owner-pet pairs, identify the most common etiologic agents, and find thelikely connection between the carrier status of an owner and the presence of dermatophytes in their pets, whether feline or canine.From May 2019 to January 2020, 21 cats and 115 dogs with their respective owners were selected for dermatophyte culture. All the dogs and cats included in the study were from thecommunities of Chilpancingo, Acalmani, and Chilapa  and were examined at their respective homes. The pairs lived in urban (Chilpancingo), suburban (Chilapa),and rural  areas. It should be mentioned that only pairs, meaning one animal one human per house, were included in this study. Furthermore, the inclusion criteria were lack of skinlesions and no history of dermatophytosis during the months before sampling.After the informed consent was obtained from the pet owners, the socio-demographic and clinical characteristics of the pairs were collected via a questionnaire.Regarding ethical considerations, the information of patients was handled according to the Helsinki declaration. The informed consent and the research were approved by theEthics Committee of the Universidad Autonoma de Guerrero (FCQB200619). Each pair was examined for detached hair and/or alopecia, peeling, and crusting. However, samples of the scalp or body of the pets were taken of all pairs, regardless of whether they had clinical symptoms or not. The samples were taken with a cotton swab, which was vigorously rubbed and twisted on the scalpor body of the pets four times. After the procedure was completed, the cotton swab was placed in 0.1% Tween. Since all the patients in the study were asymptomatic, we considered using this method as it is theleast invasive and has the same reliability as the toothbrush method  . FungalP<0.05 were considered statistically significant.A database was created with the information collected from the questionnaires and the laboratory results. The frequency analysis was carried out using the Xi-square statisticaltest and the values of In total, 136 pet-owner pairs were sampled, and dermatophytes were found in 3.67% of them which reflected a total of five strains, three of which were isolated in Chilpancingo,one in Chilapa, and one in Acalmani, without statistically significant differences. Moreover, four out of the five culture-positive cases were isolated from owners and one from a pet.Trichophyton tonsurans andone as T. verrucosum. In the micro-morphology of the two remained isolates from the skin of one owner and one pet, thin hyphae and microconidia along withhyphae were identified microscopically. However, other fungal characteristic structures were not identified and thereby two isolates were reported as Trichophyton sp. In this sense,several dermatophytes are known which do not or poorly sporulate in culture and thereby show very limited phenotypic characteristics [ According to the macroscopic and microscopic characteristics of the isolates as well as the growth on Mycosel Agar, two of the scalp isolates were identified as ristics  . P=0.010). Regarding their behaviors as pairs, in Chilpancingo, 18.31% of owners and pets always slept together, while in Chilapa, 92.5% of the pairs never did (P=0.001).Moreover, 38.03%, 12.5%, and 12% of the pet owners in Chilpancingo, Chilapa, and Acalmani always fed their pets, respectively (P=0.001).It was also observed that 78.8%, 95%, and 32% of the pets in Chilpancingo, Chilapa, Acalmani were not taken to public parks, respectively (P=0.001), compared to the other locations (P=0.001).The prevEven though the same dermatophytes were not found on the skin of pets and the scalp and hands of pet owners, it is important to describe the collected data.In a study performed by Cafarchia et al., the prevalence rates of dermatophytes in cats and dogs were found to be 28.2% and 20.5% respectively, while in our investigation thisprevalence rate was lower. The likely explanation for such a significant difference could be the low number of sampled cats (n=21) in the present study.It could also be due to the inclusion of two groups in the study conducted by Cafarchia et al.  , in oneIn addition, the prevalence is commonly high in owner-pet pairs that include symptomatic cases  . A highIn another study, Debnath et al. found that 20.93% of healthy dogs were positive for dermatophytes; however, they described that the age of the dogs and the seasonof the year can influence the frequency of dermatophytes  . RegardRegarding hand samples, there is little information available about the patients without apparent symptoms. However, such asymptomatic lesions could probably berelated to the low virulence of the strains existing in these lesions, for which enzymatic profiles of dermatophytes strains recovered from these infections could even be testedas previously described   . Likewise, the development of infection in the owner can be attributed to the characteristics of the pet-owner interaction and the duration of their contact  . Nevertwhich plays an important role in zoonotic transmission, is well known  , 15. TP= 0.001). Therefore, it could not be the factor favoring the appearance of asymptomatic dermatophyte carriers. In this study, it was found that the three studied populations had different hygiene habits and lifestyles based on their residence location, namely locality urban (Chilpancingo),peri-urban (Chilapa), and rural  areas. One example of this would be the sharing of pillows; while the pet always slept with the owner in 18.3% of cases in Chilpancingo,this activity was not common in other communities . Although the presence of dermatophytes on the scalp of apparently healthy individuals may, in some cases, be a temporary event, it is reasonable to assume thatasymptomatic carriers play a role in the spread and persistence of scalp ringworm in the community  . These T. tonsurans was determined in asymptomatic carriers. In this sense, the etiological agent has been found as specific to the asymptomatic carriers [ T. tonsurans has been reported to have displaced almost all the species of dermatophytes that cause tinea capitis in North America [ T. tonsurans plays a central role in symptomatic disease and can be recovered from hosts without symptoms (asymptomatic carriers) at high rates,comparable to the rates observed in patients with clinical infection [ Regarding etiology, the presence of arriers  , 14. IAmerica  . This mAmerica  - 24. Ffection  , 27 whT. verrucosum has also been found in asymptomatic carriers of Tinea capitis and is considered an important etiologic agent [ M. audouinii [ M. canis or T. mentagrophytes, are usually present with a symptomatic inflammatory response and are less frequently associated with a carrier status [ It should be noted that c agent  , 29 indouinii  . In con status  , 30.Trichophyton sp. It is noteworthy that two slow-growing filamentous fungi were isolated from the skin of pets and scalp of a patient on Mycosel agar with short hyphae and the presence of microconidia.However, we were unable to observe other structures, such as spiral or tendril hyphae, cigar-shaped macroconidia, or colony pigmentation; hence, they were only defined as Lifestyle, which is different in various localities (populations), can help the development of carrier state in people who have notable contact with pets.Isolation of different dermatophyte species from the skin of pet/owner pairs in this study denoted an asymptomatic carrier state in humans that cannot be related to thecoexistence of the same isolate in the pets. The authors would like to appreciate Lilia Lizette García Díaz (biotechnology) and Alberto Patricio Hernández (chemistry) for their support in the conservation of strains and all owners and their pets. Conceptualization: A.R.P. and R.A.G; methodology: M.G.R.R. and I.H.A; supervision: A.R.P.; statistical analysis: S.A.P.R and E.R.B; investigation: R.A.G and A.R.P;writing and original draft preparation: A.R.P., R.A.G, and J.T.J. All authors read and approved the final manuscript.The authors declare no conflicts of interest related to this study.This research was not funded by any program.No financial interests related to the material of this manuscript have been declared.


MTHFR) is an enzyme that plays a crucial role as a methyl-group donor in demethylation of homocysteine. The aim of this systematic review and meta-analysis was to study the relationship between MTHFR gene polymorphism and metabolic syndrome (MS). We used search engines and databases such as Science Direct, Google Scholar, Embase, Cochrane Library, and PubMed to identify eligible studies up to 2018. The articles were studied based on keywords including MTHFR, mutation, variant, and polymorphism in combination with MS. Data was analyzed using Comprehensive Meta-Analysis version 2.2.064 software. After extracting the data from seven articles, the total number of subjects was 1280 in the patient group and 1374 in the control group. The odds ratio was estimated to be 1.078 for the allele model of T vs. C , 1.157 for the allele model of CC vs. CT (95% CI: 0.829-1.615), 1.020 for the allele model of CT + TT vs. CC (95% CI: 1.611-0.646) and 0.799 for the allele model of TT vs. CC + CT (95% CI: 1.185- 0.539). As well, the results showed no statistically significant correlation between polymorphism genotypes of the MTHFR gene and MS (P<0.05). In general, this study showed that the presence of C677T polymorphism in the MTHFR gene has no effect on the incidence of MS.Methylenetetrahydrofolate reductase ( MTHFR) is an enzyme that plays an important role as a methyl-group donor in demethylation of homocysteine [MTHFR gene, thymine is replaced by cytosine, followed by the translation of valine instead of alanine in the structure of the produced enzyme, resulting in the formation of a temperature-sensitive enzyme and thus reducing its activity [MTHFR gene) with hypertension [MTHFR and metabolic syndrome (MS) in certain groups of people with type 2 DM [MTHFR and MS, and no comprehensive study has been done to summarize these outcomes in patients with MS. Regarding the high prevalence of MS and the role of genetic factors in the disease [MTHFR polymorphism and MS can be helpful. In this way, people who are genetically predisposed to the disease can be identified more quickly to fulfill preventive interventions. The aim of this systematic review and meta-analysis was to study the relationship between MTHFR gene polymorphism and MS.Methylenetetrahydrofolate reductase  to estimate odds ratio (OR), original research articles, randomized and controlled articles, and considering gene polymorphism as the main independent variable. Also, when several investigations were conducted on the same population, the latest study was selected to enter into the meta-analysis. The standard information form was used for data collection. Form information included the author name, publication year, study location, genotype type, total number of subjects in case and control groups, genotype distribution in the case and control groups, and the frequency of the dominant allele in both case and control groups. Two of the authors performed the process of extracting information from articles. Hardy-Weinberg equilibrium, genotyping methods qualification, controls source, sample size, and cases representativeness, were examined with a total score of 10 in this scoring. This checklist had been used in previous studies. Besides, scores zero to four were categorized as a weak study, five to seven as an average study, and eight to 10 as a strong article. The allele frequency for gene polymorphism was determined in each study using the allele counting method. OR with 95% CI was used to evaluate the power of correlation between MTHFR gene and MS, followed by the allele model (C vs. T), multiplicative model (CC vs. TT), the dominant model (CC + CT vs. TT), and recessive model (CC vs. CT + TT). Heterogeneity was calculated using measurement test and the random effects model. The I-square (I2) index of 25, 50, and 75 percent showed lower, moderate, and high levels of heterogeneity, respectively. The bias of published articles was examined using a funnel plot versus standard error (SE). Publication bias was evaluated using both Begg’s funnel plot and Egger’s linear regression test. Because of significant heterogeneity of the results (I2>30%), the random effects model, which takes the diversity of the studies into account, was used. All analyses were performed by Comprehensive Meta-Analysis version 2.2.064 (CMA) software. AP-value of less than 0.05 was considered as a significance level.Search engines and databases including PubMed, Embase, ScienceDirect, Cochrane Library, and Google Scholar were searched to find all English articles up to 2018. The related keywords were extracted using the Medical Subject Heading (MeSH), containing “I2 test and Q-test. Heterogeneity was observed in all the models, i.e., allele (T vs. C), homozygous (TT vs. CC), heterozygous (CT vs. CC), dominant (CT + TT vs. CC), and recessive (TT vs. CT + CC) genotype model, which were included for the meta-analysis. Therefore, the random effects model, which takes the diversity of the studies into account was used for data analysis. Meta-analysis of C677T polymorphism in the MTHFR gene and MS in total consisted of six case-control studies and one cohort study. Available studies in the meta-analysis were used to evaluation of the relationship between MTHFR polymorphism and MS. In general, these studies showed that the total number of subjects was 1280 in the MS patient group and 1374 in the healthy control group. According to the findings of this study , in the homozygous model , in the heterozygous model , in the dominant model  and in the recessive model .In total, 95 studies were systematically reviewed. Some studies were excluded because of the similarity of the studied samples (n=69), the inappropriateness of the samples and the failure to consider the MS as the main variable (n=22), and inadequate information on alleles (n=4). Finally, seven papers -23 were is study , there wMTHFR gene and MS. According to the C677T polymorphism in the MTHFR gene and MS, the result of the present study is in the same line with a study carried out by Russo et al. [MTHFR polymorphism and MS in patients with type 2 DM with mild hyperhomocysteinemia [et al. [MTHFR polymorphism and the prevalence of MS. However, Ellingrod et al. observed that the CT mutation in the MTHFR gene predisposes those with schizophrenia taking atypical antipsychotics to MS [MTHFR enzyme acts as a methyl-group donor for the remethylation of homocysteine and its conversion into methionine. Methionine consumes methyl group for DNA methylation, especially in CpG pairs. These pairs, which exist in certain regions, act as a promoter for related genes [MTHFR and MS through these epigenetic mechanisms [MTHFR mutation and subsequently affect DNA methylation and gene expression. We had some limitations in this study. These limitations included lack of access to some of the main articles in English and non-English languages.According to the findings of this study, there was no significant relationship between C677T polymorphism in the o et al. . They inteinemia . Yamada  [et al.  examinedcs to MS . This incs to MS . This geed genes . As a rechanisms . It is nMTHFR gene has no effect on the incidence of MS. It is suggested to evaluate the effect of folic acid fortification and supplementation on the expression of the MTHFR gene, in particular, those associated with chronic diseases such as hypertension, DM, and MS.In general, this study showed that the presence of C677T polymorphism in the We are sincerely thankful to our counsellors in Clinical Research Development Center of Shahid Mohammadi Hospital. The study was approved by the ethics committee of Hormozgan University of Medical Sciences (grant number: 970386 and ethics committee registration code: IR.HUMS.REC.1397.327). We would like to thank all the people who helped us with collecting the data and the searching works.None.


The field of Nanotheranostics, the namesake of this journal, lost two foundational pillars last year. It is difficult to accept that Dr. Sam Gambhir and Dr. Moritz Kircher both passed away much too soon. Two individuals who dedicated their careers to, what at the time they started, was radical new research fusing molecular imaging, therapy and nanoscale design. Both were founding editorial board members of Nanotheranostics. Dr. Kircher was the editor in chief of Nanotheranostics at the time of his passing. These physician scientists were motivated to translate this type of innovative basic research into their clinics in order to benefit humankind.Stanford Radiology, where both spent significant time, represents an intellectual headquarter and origin for much of the theranostics field. Indeed, it is difficult to imagine research on the topic not being strongly influenced by work carried out by scientists who spent time there. It should be sadly noted that in 2018, Stanford radiologist Dr. Jürgen Willmann was lost in a car accident at age 45. The string of untimely departures is a terrible coincidence that, besides personal loss for friends and family, significantly sets back progress to nanotheranostics. These physician scientists were pioneering a brand of human health research that was multidisciplinary, clinically-minded, and highly innovative. It is unclear when their roles can be filled in the future by other researchers as driven and talented as they were.It was therefore with mixed emotions that I assumed the role of Editor-in-Chief of Nanotheranostics last year. I will try to follow in the editorial footsteps of Dr. Kircher. It should also be said that despite the setbacks, there are tremendously exciting developments with respect to the current state of the field, that provide grounds to be optimistic about the future. Nanotheranostics, as a journal, will continue to publish cutting edge research in the field.This special memorial issue aims to honor the legacy of Dr. Gambhir and Dr. Kircher. It features articles contributed posthumously by both Dr. Gambhir, Dr. Kircher, and also the late Dr. Mike Detty, who made contributions to the field of Raman spectroscopy dyes. Enormous thanks go to all authors, and the corresponding contributors to this issue, including Dr. Stefan Harsen, Dr. Ramasamy Paulmurugan, Dr. Jesse V. Jokerst, Dr. Chrysafis Andreou, Dr. Jeesu Kim, Dr. Peng Huang, Dr. Xiaoyuan Chen, Dr. Carina Mari Aparici and Dr. Natesh Parusharama.


Following publication of the original article , one of The email address of the corresponding author, Abbas Alemzadeh, is alemzadeh@shirazu.ac.ir.The original article  has been


Impatiens longiaristata , a new species from western Sichuan Province in China, is described and illustrated here based on morphological and molecular data. It is similar to I. longiloba and I. siculifer, but differs in its lower sepal with a long arista at the apex of the mouth, spur curved downward or circinate, and lower petal that is oblong-elliptic and two times longer than the upper petal. Molecular analysis confirmed its placement in sect. Racemosae. Simultaneously, during the inspection of the protologues and type specimens of allied species, it was found that the types of two names from this section were syntypes based on Article 9.6 of the International Code of Nomenclature for algae, fungi, and plants (Shenzhen Code). According to Articles 8.1, 9.3, and Recommendations 9A.1, 9A.2, and 9A.3, the lectotypes of these two names are here designated. Impatiens L.  is one of the most species-rich genera of angiosperms with more than 1000 species , , and . A critical examination of digital images from all collections reveals that the Ducloux, F. 532 digital image registered Henry, A. as the collector on the mounted herbarium sheets in Kew. In addition, this collection was cited at the Museum d’Histoire Naturelle herbarium but could not be traced among online digital images under Ducloux, F. 532 collections in P but was rather found in the Kew digital herbarium. It is not clear whether at one point herbarium specimens were transferred from P to K or the digital images were obtained by the Royal Botanical Gardens, Kew during databasing and digitization sessions of 185,937 specimens at P  from Yunnan, Mangtze ,  from Yunnanfu ,  from Yunnan, the divide between the Mekong and Shang valley on Jung-yuch-Talifu road  and  from Yunnan. Upper Burma, N.W. Yunnan Ming Kwang divide . All examined collections of digital images had hand drawings of the plant reproductive illustrations . Furthermore, 1004 and 1006 had accompanying comprehensive handwritten fieldnotes that matched the original description. However, after a critical examination of all these syntypes, we found the specimen displaying the best representative material and locality information (including elevation), numbered 9762, best fitted the original portal description and was preserved in the Kew herbarium. Moreover, this specimen retains the original pigmentation, making the collections more appealing and providing a precise representation of the living plant species. Hence, we designate it here as the lectotype  in I. siculifer and apiculate in I. longiloba. The spur is curved downward or circinate in I. longiaristata, while it is curved upward in I. siculifer and slightly curved at the tip in I. longiloba. The lateral united petals of I. longiaristata are ca. 3 cm long, the upper petal is triangular and semi-circular, and the lower petal is oblong-elliptic, tip rounded, ca. 1.8 cm long, and two times longer than the upper petal, whereas in I. siculifer the lateral united petals are ca. 1.8 cm long, the upper petal is subtriangular, and the lower petal is lorate with an acuminate tip, and in I. longiloba the lateral united petals are ca. 2.5 cm long and the lower petal is lorate, elongate, and four times longer than the upper petal. The lateral veins of leaves are 5–7 pairs in I. longiaristata, but they are 10–12 pairs in I. longiloba and 5–11 pairs in I. siculifer. The lateral sepals are two pairs in I. longiaristata and I. longiloba, but just a pair in I. siculifer. The detailed morphological comparisons between new species and allied species are shown in I. longiaristata and I. longiloba formed a highly supported clade (PP = 1.00 and BP = 100) with I. siculifer forming a sister clade. It was also noted that I. longiaristata was morphologically closer to I. longiloba than to I. siculifer (st China ,10 and Vst China . Morphold petals . The lowiculifer . Impatiens longiaristata is a new taxon in science. Detailed descriptions, color plates, phylogenetic analysis, and comparisons with species of similar taxa are provided. The lectotypes of I. aquatilis and I. clavicuspis are also designated.Based on detailed morphological studies and phylogenetic analysis,


In the original article, there were two errors.(1) Our information on Wnt modification and secretion was out of date. Mouse Wnts are not palmitoleated on cysteines—that was an error in mutational analysis by Karl Willert. All the cysteines in Wnt are engaged in disulfide bonds . However, a new reference describes WNT palmitoylation in zebrafish WNT3A .(2) The WLS protein binds to Wnt in the ER, not the Golgi. The Golgi localization of WLS was also an error due to the use of an epitope tag on the c-terminus .the introduction, paragraph number 2A correction has been made to The WNT family now contains 19 WNT genes, falling into 12 WNT subfamilies in mammalian genomes. All WNT genes encode proteins around 40 kDa in size and contain highly conserved cysteines Miller, . MammaliAccordingly, the following reference has been added to the original article:Front. Cell Dev. Biol. 9:671218. 10.3389/fcell.2021.671218Dhasmana, D., Veerapathiran, S., Azbazdar, Y., Nelanuthala, A. V. S., Teh, C., Ozhan, G., et al. (2021). Wnt3 is lipidated at conserved cysteine and serine residues in zebrafish neural tissue. And the following reference has been removed from the original article:Nature 423, 448–452.Willert, K., Brown, J. D., Danenberg, E., Duncan, A. W., Weissman, I. L., Reya, T., et al. (2003). Wnt proteins are lipid-modified and can act as stem cell growth factors. The authors apologize for these errors and state that this does not change the scientific conclusions of the article in any way. The original article has5 been updated.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.


Sargus bipunctatus , feeding on a human corpse. The morphology of collected imagines and larvae of S. bipunctatus was documented, and a standard COI barcode sequence was obtained. Morphology- and DNA-based methods were used to distinguish the larvae of S. bipunctatus and its relative, Hermetia illucens . The potential of S. bipunctatus for practical applications in forensic entomology is currently difficult to assess.In the current study, we present the first record of twin-spot centurion fly larvae, Hermetia illucens . Analysis of fauna found on a human corpse, discovered in central Poland, revealed the presence of feeding larvae of another species from this family: the twin-spot centurion fly Sargus bipunctatus . The investigated corpse was in a stage of advanced decomposition. The larvae were mainly observed in the adipocere formed on the back and lower limbs of the corpse, and in the mixture of litter and lumps of adipocere located under the corpse. Adult specimens and larvae were identified based on morphological characters, and final identification was confirmed using DNA barcoding. Implementing a combination of morphological and molecular methods provided a reliable way for distinguishing the larvae of S. bipunctatus and H. illucens. The potential of S. bipunctatus for practical applications in forensic entomology is currently difficult to assess. Wide and reliable use of S. bipunctatus in the practice of forensic entomology requires further studies of the bionomy of this fly.The only European Stratiomyidae species known for feeding on human corpses was the black soldier fly  The family Stratiomyidae (soldier flies), representing the suborder of Brachycera Orthorrhapha, includes more than 2600 described species . Large sHermetia illucens . This species, probably native to Central America, can currently be found in warmer regions around the world. In the northern part of North America, it reaches the province of Ontario , is widespread in Europe and the northwest of North America [Dociostaurus moroccanus , and Oldroyd [Cerioporus squamosus (Huds.) Quélet (1886). The variety of substrates from which larvae of S. bipunctatus has been collected indicates a broad feeding spectrum. Despite this, only Chick [S. bipunctatus and decaying animal remains. Numerous females of the species were observed on the carcass of a domestic pig on the 24th day of decomposition. Nineteen days later, larvae were found feeding in a mixture of soil, mulch, and the putrefactive liquid exuding from the carcass. Sukontason et al. [Sargus sp. larvae feeding on human bodies found in the forests of Thailand from 2000–2006.The twin-spot centurion fly, Scopoli, 763 [=Chr America ,17. Sing America ,20. Acco America , it is a America ,22,23,24 America , larvae  Oldroyd  recordedly Chick  has repon et al.  recordedS. bipunctatus are very distinctive and easy to distinguish from the specimens of other necrophagous flies by their dorsoventral flattening. The only species possible to misidentify it with is Hermetia illucens, which belongs to the same family Stratiomyidae. The geographic distributions of both species overlap [H. illucens will expand through both natural processes and the unintentional release of mature flies from black soldier fly breeding farms. Larvae of S. bipunctatus and H. illucens can be easily distinguished based on morphological features alone. Sargus bipunctatus has a distinct colouration pattern, with six longitudinal stripes on its abdominal segments, and very short dorsal, dorsolateral, lateral and ventral bristles , included in the red lists of threatened animals in Central European countries [N. littoralis development, conducted under laboratory conditions [S. bipunctatus. Further studies of this species should have a precise forensic profile and cover field studies of environmental preferences, preferred food sources and habitats, role in necrophagous insect community, and activity period during successional changes of carrion, as well as laboratory experiments concerning thermal requirements during the development of immature life stages.Imagines of  October . In the albiceps . More detomology . Based oountries ,39. Analountries ,40. Suchnditions ,42. Finanditions ,44. We h