Automated identification of aquatic insects: A case study using deep learning and computer vision techniques.

Despite huge attention in other domains, deep learning is only very slowly beginning to be applied in biodiversity research. Mayflies (Ephemeroptera), stoneflies (Plecoptera) and caddisflies (Trichoptera), often abbreviated as EPT, are frequently used for freshwater biomonitoring due to their large numbers and sensitivity to environmental changes. However, the unambiguous morphological identification of EPT species is a challenging, but fundamental task. Morphological identification of these freshwater insects is therefore not only extremely time-consuming and costly, but also often leads to misjudgments or generates datasets with low taxonomic resolution. Here, we investigated the application of deep learning to increase the efficiency and taxonomic resolution of biomonitoring programs. Our database contains 90 EPT taxa (genus or species level), with the number of images per category ranging from 21 to 300 (16,650 in total). Upon completion of training, a CNN (Convolutional Neural Network) model was created, capable of automatically classifying these taxa into their appropriate taxonomic categories with an accuracy of 98.7 %. For the extensive set of 68 tested taxa, our model achieved a perfect classification rate of 100 %. We achieved noteworthy classification accuracy with morphologically closely related taxa within the training data (e.g., species of the genus Baetis, Hydropsyche, Perla). Gradient-weighted Class Activation Mapping (Grad-CAM) visualized the morphological features responsible for the classification of the treated species in the CNN models. In the Ephemeroptera, the head was the most important feature, while the thorax and abdomen were equally important for the classification of Plecoptera taxa. For the order Trichoptera, the head and thorax were almost equally important. Our database is recognized as the most extensive aquatic insect database, notably distinguished by its wealth of included categories (taxa). Our approach can help solve long-standing challenges in biodiversity research and address pressing issues in monitoring programs by saving time in sample and data processing.

Relating environmental pressures to littoral biological water quality indicators in Western Balkan lakes: Can we fill the largest gaps?

Along six transects in each of six lakes across the Western Balkans, we collected data for three groups of littoral biological water quality indicators: epilithic diatoms, macrophytes, and benthic invertebrates. We assessed the relationships between them and three environmental pressures: nutrient load (eutrophication), hydro-morphological alteration of the shoreline, and water level variation, separating the effect of individual lakes and continuous explanatory variables. Lake water total phosphorus concentration (TP) showed substantial variation but was not related to any of the tested biological indicators, nor to any of the tested pressures. We suggest that this may be due to feedback processes such as P removal in the lake littoral zone. Instead, we found that a gradient in surrounding land-use towards increasing urbanization, and a land-use-based estimate of P run-off, served as a better descriptor of eutrophication. Overall, eutrophication and water level fluctuation were most important for explaining variation in the assessed indicators, whereas shoreline hydro-morphological alteration was less important. Diatom indicators were most responsive to all three pressures, whereas macrophyte biomass and species number responded only to water level fluctuation. The Trophic Diatom Index for Lakes (TDIL) was negatively related to urbanization and wave exposure. This indicates that it is a suitable indicator for pressures related to urbanization, although a confounding effect of wave exposure is possible. Invertebrate abundance responded strongly to eutrophication, but the indicator based on taxonomic composition (Average Score Per Taxon) did not. Our results suggest that our metrics can be applied in Western Balkan lakes, despite the high number of endemic species present in some of these lakes. We argue that local water management should focus on abating the causes of eutrophication and water level fluctuation, whilst preserving sufficient lengths of undeveloped shoreline to ensure good water quality in the long run.

Burnout of Physicians, Pharmacists and Nurses in the Course of the COVID-19 Pandemic: A Serbian Cross-Sectional Questionnaire Study.

This research was a prospective, cross-sectional observational study of 128 health workers in the central part of the Republic of Serbia. The study surveyed health workers (physicians, pharmacists and nurses) who worked during peaks of the COVID-19 pandemic in the Republic of Serbia in June and November 2020. The Maslach Burnout Survey for Medical Personnel addresses three scales: (a) emotional exhaustion (EE) measures feelings of being emotionally overextended and exhausted by one's work; (b) depersonalization (DP) measures an unfeeling and impersonal response toward recipients of one's service, care treatment, or instruction; and (c) personal accomplishment (PA) measures feelings of competence and successful achievement in one's work. Linear regression and the chi-square test were used to test a relationship between the input variables (x) and the single output variable (y). We can conclude that most health workers had a high degree of emotional exhaustion, but also a low degree of depersonalization and a high degree of sense of personal achievement. Nurses and physicians had similar answers on the pandemic during their work, but pharmacists had different answers.

The Rhyacophila fasciata Group in Europe: Rhyacophila fasciata Hagen 1859 and formerly synonymized species (Trichoptera: Rhyacophilidae), with new description of Rhyacophila fasciata and Rhyacophila septentrionis McLachlan 1865 (stat. prom.).

The presence and distribution of Rhyacophila fasciata Hagen 1859 in Europe were revised, based on bibliographic study, collection specimens, and new material collected in different countries. The status of formerly synonymized species, Rhyacophila ferruginea (Scopoli 1763) and Rhyacophila septentrionis McLachlan 1865 was also assessed. The type of R. ferruginea is missing, the taxon is still unidentified, and thus we propose Rhyacophila ferruginea as a nomen dubium. Morphological features and genetic evidence revealed that R. septentrionis differs from R. fasciata, so we propose to change its status to status resurrectus. We therefore include new descriptions of the different stages (larva, pupa, male, and female) of R. fasciata and of R. septentrionis, together with a molecular analysis based on mitochondrial cytochrome oxidase I (mtCOI) and ecological notes. The species R. coppai Oláh 2020 NEW SYNONYM and R. soreda Coppa Oláh 2020 NEW SYNONYM are synonyms of R. sociata Navás 1916; the species R. kopasa Oláh Coppa 2020 NEW SYNONYM and R. rova Oláh Coppa 2020 NEW SYNONYM are synonyms of R. denticulata McLachlan 1879; the species R. matrensis Oláh Szczesny 2020 is probably a synonym of R. fasciata, so more study of this species is needed in order to confirm or deny that it is a valid species.

Littoral eutrophication indicators are more closely related to nearshore land use than to water nutrient concentrations: A critical evaluation of stressor-response relationships.

Biological assessment metrics and water chemistry measurements are used to quantify the link between stressors and their effects on lake ecosystems, for the Water Framework Directive. However, correlations between metrics and water chemistry are often poor. This is seen as major weaknesses of Water Framework Directive-related monitoring and assessment. We analyzed macrophytes, benthic algae, benthic macroinvertebrates, water chemistry and sediment total phosphorus content in the littoral of six lakes in the Western Balkans and used CORINE land use data to estimate nutrient enrichment via runoff from the adjacent land. Lakes with a higher estimated phosphorus runoff from the adjacent land did not have higher littoral water nutrient concentrations, but littoral diatom assemblages indicated more eutrophic conditions. These lakes also had higher abundances of littoral benthic primary producers, which in turn were associated with low concentrations of dissolved nutrients, but only in autumn, not in spring. This is consistent with primary producers taking up nutrients during the summer growth season. In lakes with high abundances of benthic primary producers, it is likely that the littoral vegetation plays a large role in the transfer of nutrients from the water to the benthos. This process impairs correlations between biological metrics and water nutrient concentrations. Our results suggest that CORINE land cover may be more useful to characterize littoral nutrient enrichment than lake water chemistry. Increased benthic primary producer biomasses and "eutrophic" diatom indices may indicate littoral nutrient enrichment even if water nutrient concentrations are low.

The response of chironomid taxonomy- and functional trait-based metrics to fish farm effluent pollution in lotic systems.

The lotic habitats affected by trout farm waste are colonized with a particular invertebrate community of which chironomids are the most abundant group. However, there is little information available regarding how chironomid community structures respond to this type of pollution at the highest taxonomic resolution. Eight fish farms, together with their lotic systems as recipients, were used to test the variability of the chironomid community and its surrogates (taxonomic and functional metrics) across spatially arranged sampling sites to form a gradual decrease in the trout farm influence. The self organizing map (SOM) classified six different types of chironomid communities which were characteristic for both the control and affected habitats. The species indicator analyses listed 32 taxa as positive indicators of water pollution. The SOM and Kruskal-Wallis test revealed that the pattern of chironomid community structure obtained was mainly driven by six environmental parameters (Altitude, conductivity, distance from the outlet, hardness, HN4-N, NO3-N). Categorical principal components analysis (CATPCA) derived three models for each type of biotic metric, in which for diversity-, taxonomy- and functional feeding group-based metrics, the first two dimensions explained 55.2%, 58.3% and 55.4%, of the total variance respectively for 315 sampling sites. According to this analysis, the total number of taxa (S), abundance and the Shannon-Wiener index (H') (as a diversity metric), as well as the proportion of Tanypodinae (as taxonomic group) and grazers/scraper (GRA) and gatherer collector (GAT)(as FFG metrics), were related to the outlet distance gradient, thus showing great potential to be used in the multimetric approach in bioassessment.

Detectability of fentanyl and designer fentanyls in urine by 3 commercial fentanyl immunoassays.

In recent times, structural variants of fentanyl (designer fentanyls) have appeared on the recreational drug market for new psychoactive substances (NPS). These potent opioids have caused harmful intoxications and increased opioid-related mortality in many countries. This work evaluated 3 commercial immunoassays for fentanyl screening in urine and investigated whether they are useful also for screening of designer fentanyls. The assays examined were the Thermo DRI® Fentanyl Enzyme Immunoassay, the ARK™ Fentanyl Assay homogeneous enzyme immunoassay, and the Immunalysis® Fentanyl Urine SEFRIA™ Drug Screening Kit. A liquid chromatography-high-resolution mass spectrometry method was used as reference. The DRI fentanyl immunoassay generated somewhat higher assay imprecision values (%CV) compared with the ARK™ and SEFRIA™ assays, but all assays showed %CV values acceptable for routine use. The 3 assays showed overall good detectability (33%-95% cross-reactivity) for blank urine samples spiked with acetylfentanyl, acrylfentanyl, butyrfentanyl, 4-chloroisobutyrfentanyl, 4-fluorobutyrfentanyl, 4-fluorofentanyl, 4-fluoroisobutyrfentnyl, isobutyrfentanyl, methoxyacetylfentanyl, or tetrahydrofuranfentanyl, whereas 4-methoxybutyrfentanyl (all assays) and 2-fluorofentanyl (DRI assay) showed low cross-reactivity. A good detectability of designer fentanyls was confirmed in urine samples from authentic acute intoxications. In conclusion, the present results demonstrate that the urinary fentanyl immunoassays are generally useful also for preliminary screening of fentanyl analogs sold as NPS. When the SEFRIA™ assay was applied for testing of 980 urine samples from patients treated for drug dependence in Sweden, only 1 sample was confirmed positive for fentanyl.

Integrative taxonomy by molecular species delimitation: multi-locus data corroborate a new species of Balkan Drusinae micro-endemics.

BACKGROUND: Taxonomy offers precise species identification and delimitation and thus provides basic information for biological research, e.g. through assessment of species richness. The importance of molecular taxonomy, i.e., the identification and delimitation of taxa based on molecular markers, has increased in the past decade. Recently developed exploratory tools now allow estimating species-level diversity in multi-locus molecular datasets. RESULTS: Here we use molecular species delimitation tools that either quantify differences in intra- and interspecific variability of loci, or divergence times within and between species, or perform coalescent species tree inference to estimate species-level entities in molecular genetic datasets. We benchmark results from these methods against 14 morphologically readily differentiable species of a well-defined subgroup of the diverse Drusinae subfamily (Trichoptera, Limnephilidae). Using a 3798 bp (6 loci) molecular data set we aim to corroborate a geographically isolated new species by integrating comparative morphological studies and molecular taxonomy. CONCLUSIONS: Our results indicate that only multi-locus species delimitation provides taxonomically relevant information. The data further corroborate the new species Drusus zivici sp. nov. We provide differential diagnostic characters and describe the male, female and larva of this new species and discuss diversity patterns of Drusinae in the Balkans. We further discuss potential and significance of molecular species delimitation. Finally we argue that enhancing collaborative integrative taxonomy will accelerate assessment of global diversity and completion of reference libraries for applied fields, e.g., conservation and biomonitoring.

Morphological features of larvae of Drusus plicatus Radovanovic (Insecta, Trichoptera) from the Republic of Macedonia with molecular, ecological, ethological, and distributional notes.

A description of the larva of Drusus plicatus Radovanovic is given for the first time. The most important diagnostic characters enabling separation from larvae of the other Drusinae from the southeast Europe are listed. Molecular, ecological, and ethological features and distribution patterns of the species are given. Additionally, information on the sympatric caddisfly species of the three springs where larvae and adults of Drusus plicatus were found and presented.

The trout farm effect on Dinocras megacephala (Plecoptera: Perlidae) larvae: Antioxidative defense.

Trout production represents a major agricultural activity in Serbia. Organic compounds are drained into the environment, usually without previous sedimentation, thus affecting the stream biota. Considering that biological monitoring is commonly based on aquatic macroinvertebrates as target organisms, the authors used larvae of Dinocras megacephala to estimate trout farm effects on the mass of the larvae and their antioxidative defense in pollution stress conditions. Four locations were chosen along the channel of the Raska River, 2 upstream (L1, L2) and 2 downstream (L3, L4) from the trout farm outlet. Basic physical and chemical water parameters were measured. Superoxide dismutase (SOD) and catalase (CAT) activity changes were analyzed to determine the level of oxidative stress caused by the increase of organic compounds originating from the trout farm. Dissolved oxygen concentration decreased from the upstream to downstream locations. Furthermore, the concentration of ionized ammonia was almost 10 times higher at the downstream locations than at the upstream locations. Larval mass, as well as CAT activity, was significantly higher at L3 compared with the other 3 locations. Activity of SOD was significantly higher at L3 than at L1. The results indicate that higher concentrations of organic compounds from the trout farm induce clear changes in the status of the antioxidant defense of D. megacephala larvae. Environ Toxicol Chem 2016;35:1775-1782. © 2015 SETAC.

Nature and results of treatment of war wounds caused by cluster bombs.

The aim of this study is to describe the nature of war wounds with fracture caused by cluster bombs and to suggest treatment options for such injuries. The nature of wounds caused by cluster bombs differs from those caused by conventional arms (they are more severe). The sides of the wounds are represented by conquasated soft tissues (such as fat and muscle) with thick dead tissues, ordinarily with a thickness of 0.5-4.5 cm. Another main characteristic of such injuries is the high percentage of amputations needed due to the high rate of neurovascular damage. This paper investigates the cases of 81 patients who sustained a total of 99 war wounds with fractures. The average age of the patients was 32.7 years while the youngest was 20 and the oldest, 77. According to The International Committee of the Red Cross (ICRC) classification of war wounds, 14 patients had grade I injuries, 48 patients grade II, and 29 patients, grade III. Mitkovic external fixation system, known also as the "War Fixator" was used for all fractures fixation. One protocol, which was a modification of the ICRC's protocol adapted to our specific conditions, was used throughout the study. For solving soft tissue defects, a rotator fasciocutan flap was the most frequently used. For solving of bones defect Mitkovic reconstructive external fixation device was used. All fractures we treated healed. We concluded that shortening the procedural time and being a very simple, immediate using of Mitkovic versatile external fixator ("War Fixator") is, leads to desirable results.

[Knee angular deformity correction using unicorticotomy and callus distraction by external fixation method and presentation of new external fixation device for varus correction of proximal tibia].

The main goal of this paper is to present clinical results of surgical correction of angular (varus and valgus) deformities of knee area. Goal is also to present new external fixation device for surgical treatment of varus deformity of proximal tibia. As material it has been used series of 70 patients treated during the period of 6.5 years in University Orthopaedic and traumatology clinic. Biomechanical investigation of 80% cut bone model fixed by new external fixation device was performed in the measurement laboratory of Mechanical Faculty University of Nis. Clinical results of treatment of 58 high tibial surgeries (49 varus and 9 valgus) and 12 distal femur surgeries (2 varus and 10 valgus) showed that unicorticotomy and callus distraction by the use of Mitkovic external fixation device leaded to correction and bone union. Superficial pin tract infection happened in 9 patients and has been successfully treated without pins removal. Biomechanical investigation of new device showed that during axial loading of bone model by 690 N (70 kg) movement of medial distal end of bone model was 0.30 mm while posterior distal end of bone model was 0.26 mm. From results obtained in can be concluded that unicorticotomy and gradual callus distraction by the use of external fixation is one reliable method. Biomechanical testing of new external fixation device showed that it is more stable in control of varus and posterior position of proximal tibial fragment.