bluuebunny/crossref_metadata_2025 huggingface dataset:
Stigma, Shame, and Fear: Navigating Obstacles to Peace in Mindanao
Authors: Ingram Haroro
Published: March 2021

After decades of cyclical peace agreement failures and war in Mindanao, the establishment of the Bangsamoro Autonomous Region of Muslim Mindanao (BARMM) in early 2019 brought the best hope for sustainable peace and stability in living memory. But the problems within the BARMM and Mindanao more broadly are immense. A trifecta of stigma, shame, and fear is regularly identified as levers exploited by peace spoilers to not only recruit and mobilize from local communities but obstruct disengagement and reintegration efforts. The widespread and intergenerational experiences of trauma across Mindanao hang like an invisible pall over almost every aspect of life. The dynamics of stigma, shame, and fear in Mindanao tend to be multidimensional in that they may emerge from a range of sources and multidirectional in their effect, as different sources of stigma, shame, and fear can push and pull individuals and groups in different ways. Stigma, shame, and fear may act as obstacles but also opportunities that need to be understood and appropriately harnessed in disengagement and reintegration initiatives. This policy note offers a framework of recommendations that are largely grounded in peacebuilding approaches.
Choose Wisely: Metered Dose Inhalers Versus Nebulized Inhaled Solutions in The Inpatient Setting
Authors: Alshamali Meera , Lee Robin , Change Alice , Wong Johnny
Published: March 2021

Background
Healthcare expenditures continue to rise, and hospitalization costs remain the driving factor for nationally increasing health expenditures.(1,3) Affordability initiatives targeted at minimizing inpatient drug spending and medication waste are vital. Metered dose-inhalers (MDIs) impose high potential for medication waste as their bulk nature provides a day supply that far exceeds average hospitalization length of stay.(1,7) Kaiser Permanente Baldwin Park Medical Center inpatient pharmacy adopted a cost saving initiative aimed at converting the commonly prescribed ipratropium MDI to its equivalent nebulized solution.
Methods
This was a retrospective medication use evaluation (MUE), assessing the overall drug cost impact of converting patients from ipratropium MDI to ipratropium nebulized solution. The study period consisted of two phases: the pre-implementation phase from 11/1/2017 to 09/30/2018, and the post-implementation phase from 11/01/2018 to 09/30/2019.The study population consisted of patients admitted to the Critical Care Unit (CCU) and Step-Down Unit (SDU), who were prescribed an ipratropium MDI or ipratropium nebulized solution. Patients who were under the age of 18 years old were excluded. The primary outcome was to analyze the drug cost savings of prescribing nebulized solutions over MDIs in the inpatient setting. The secondary outcome was to assess overall compliance with the conversion protocol.
Results
Ipratropium MDI administrations declined by 91% (from 3,751 to 336) during the post-implementation phase. An estimated drug cost savings of $68,700 occurred by converting the ipratropium MDIs to ipratropium nebulized solution.
Conclusion
This conversion protocol was successful at reducing MDI waste and attaining substantial drug cost savings at our medical center over a period of 1 year since implementation.
The influence of functional ingredients on the technological properties of water-swelling rubber sealing elements
Authors:   Chuvash State University  of I.N. Ulyanov, Egorov Evgeny N. , Ushmarin Nikolay F. ,   Chuvash State University  of I.N. Ulyanov, Efimov Konstantin V. ,   Chuvash State University  of I.N. Ulyanov, Sandalov Sergey I. ,   Chuvash State University  of I.N. Ulyanov, Spiridonov Ivan S. ,   Chuvash State University  of I.N. Ulyanov, Koltsov Nikolay I. ,   Chuvash State University  of I.N. Ulyanov
Published: March 2021

The article investigated the influence of the nature and content of coutchoucs, sevilens, vulcanizing groups, fillers, plasticizers, ingredients of directional action on the technological properties (plasticity, ring modulus, density, start time of vulcanization, stickiness) of two rubber mixtures. These indicators characterize the processability of rubber compounds in the manufacture of products from them. The study was conducted in order to select the basis of rubber compounds for the manufacture of the outer and inner layers of water-swellable sealing elements (WSSE). It has been established that rubber compounds for the outer layer based on butadiene-nitrile BNKS-18AMN, isoprene SKI-3 and ethylene-propylene SKEPT-40 coutchoucs, as well as the inner layer of WSSE based on butadiene-nitrile BNKS-18AMN, butadiene methylstyrene SKMS-30ARK and ethylene-propylene SKEPT-40 rubbers possess satisfactory technological properties. It was shown that these rubber mixtures which contain sevilen 11808-340, a vulcanizing group (sulfur + thiazole 2 MBS + guanid F), a combination of fillers (talc + grew 175 + Karelite MK), petroleum resin Sibplast, directional ingredients (vermiculite + needle punched cloth “Oksipan”), hydrosorption additives (polyacrylamide AK 639 + sodium polyacrylate), have improved technological properties. These rubber mixtures can be recommended as the basis for the manufacture of the outer and inner layers of water-swelling sealing elements for the oil and gas industry.
Disposal and disinfection of organic waste of poultry farms
Authors:   Chelyabinsk State University, Sukharev Yury I. , Apalikova Inna Yu. ,   FGKVOU VPO Federal State  Official Military Educational Institution of Higher Professional Education Military Educational-Research Centre of Air Force «Air Force Academy named After Professor N.E. Zhukovsky and Y.A. Gagarin» (Voronezh) of the Ministry of Defence of the Russian Federation (Branch, Chelybinsk), Apalikov Vitaly O. ,   Federal state Autonomous educational institution of higher education "South Ural State  University (national research University)"
Published: March 2021

Huge amounts of litter for a number of reasons, accumulated near poultry farms, have become the object of close attention of environmental and Supervisory authorities. Almost all poultry farms of the Russian Federation were in a difficult environmental situation, as the accumulated bird droppings has become a serious source of environmental pollution, because for the disposal of such volumes of bird droppings poultry farms today do not have even the simplest sets of equipment. The emerging negative trend can lead in the very near future to an ecological disaster of farms with unpredictable negative consequences for the inhabitants of settlements, to the death of flora and fauna not only of poultry, but also of neighboring territories, it is quite possible the emergence of infectious and invasive diseases in humans, animals and birds. Chelyabinsk region is famous all over Russia for its livestock enterprises, one of the main directions of which is poultry farming. The region is one of the main suppliers of poultry products to the nearby regions of the country. With large-scale breeding of chickens, a large amount of food waste is formed, the leading of which is chicken manure. The paper presents nanotechnology reception processing of a chicken dung, based on experimental data, the destruction of the eggs of Giardia, opistorhoz, ascarids in the Indus-Tserovani stochastic resonance nanocluster current splashes, with the passage of the organic colloidal system between carbon and graphite electrodes. The paper presents a project of a semi-industrial installation for the disinfection of chicken manure, followed by its use as an effective collodic agricultural fertilizer without the use of heat treatment.
Study of dielectric properties of 4-methylstyrene and α-methylstyrene copolymer films with different molar ratio of initial monomers
Authors:   Samara National Research University, Dolgin Ignat S. , Purygin Pyotr P. ,   Samara National Research University, Zarubin Yury P. ,   Samara National Research University
Published: March 2021

Two new copolymers of 4-methylstyrene and α-methylstyrene were obtained by the emulsion polymerization method according to the previously described and tested method in the course of the experiment in laboratory conditions. The molar ratio of the starting monomers of 4-methylstyrene and α-methylstyrene was 9: 2 and 10: 1, respectively, in each material sample. The structure of the copolymers obtained was confirmed by IR and 1H NMR spectroscopy. Under laboratory conditions, three prototypes of each copolymer film were obtained from the solution. A sample of the copolymer was dissolved in methylene chloride was applied on a smooth glass substrate. After complete evaporation of the solvent, the film was separated from the substrate. The thickness of the films for each copolymer sample was 20, 30, and 50 microns. In the course of further experimentation, these samples were examined for the values of dielectric constant and tangent of dielectric loss angle. The basis of this experiment used a resonant method of measurement. The presented method is based on the variation of conductivity due to a change in the electrical capacitance of the oscillating circuit. In this experiment, the magnitude of the electrical capacitance and the tangent of the dielectric loss angle were measured. Based on the value of the capacitance and the parameters of the film sample to be measured, it is possible to calculate the dielectric constant. From the data obtained it can be seen that with increasing film thickness, dielectric constant values decrease. However, the indicators of the tangent of dielectric loss angle increase. With an increase in the measurement frequency, a slight decrease in the dielectric constant is observed, and the value of the tangent of the dielectric loss angle decreases nonlinearly. In addition, it should be noted that the samples of 4-methylstyrene-α-methylstyrene copolymer films with a molar ratio of starting monomers of 10: 1 have a higher dielectric constant, along with lower values of the dielectric loss tangent. From this it follows that samples with a lower proportion of α-methylstyrene in their structure have better dielectric characteristics compared with their counterparts with a high content of α-methylstyrene.
Measurement of volume resistivity in a series of copolymers of 4-methylstyrene-α-methylstyrene and 4-methoxystyrene-α-methylstyrene
Authors:   Samara National Research University, Dolgin Ignat S. , Purygin Pyotr P. ,   Samara National Research University, Zarubin Yury P. ,   Samara National Research University
Published: March 2021

In the course of previous experiments, a technique was developed for the preparation of a copolymer of styrene and α-methylstyrene. This technique is an emulsion polymerization in the initiator of ammonium persulfate in an inert zone of direct current argon. It was found that the best emulsifier is potassium stearate. This substance can be explained by those copolymers that showed the best dielectric performance. In the course of the new experiment, a series of new copolymers of 4-methylstyrene-α-methylstyrene and 4-methoxystyrene-α-methyl-styrene was obtained in the molar ratios of the initial monomers of 8 : 3, 9 : 2 and 10 : 1 for both rows of copolymers. The outputs of the pure product for all compounds accounted for more than 60% in terms of the initial monomers. To further measure the value of the specific volume resistance of the material, films of each newly synthesized copolymer were obtained. Samples of the copolymers were dissolved in methylene chloride and applied to a smooth glass substrate. The choice of this solvent is due to its low toxicity and low boiling point. For the experiment were selected samples with a film thickness of 50 microns. Measurements of the specific volume resistance were carried out at the Research Institute "Girikond" (St. Petersburg) using an Agilent 4339B instrument. The operating voltage was 100 V. The measurement results showed that samples of 4-methoxystyrene-α-methylstyrene copolymer were several orders of magnitude superior to samples of 4-methylstyrene-α-methylstyrene copolymer. Most likely in this case such a difference in the indices of the specific volume resistance of the copolymer films is due to the nature of the functional groups of substituents in the copolymer. From the obtained measurement results it can be seen that the samples containing the methoxy group have a high resistance value compared to the samples containing a methyl group in their structure. In a series of samples of copolymer films, an increase in the specific volume resistivity index is observed with a decrease in the α-methylstyrene content in the copolymer structure. This dependence is observed for copolymers of 4-methylstyrene-α-methylstyrene and for 4-methoxystyrene-α-methylstyrene. The sample of 4-methoxystyrene-α-methylstyrene with a molar ratio of initial monomers of 10 : 1 has the highest value of specific volume resistance among all the obtained samples of copolymer films.
On the issue of standardization of dry extract "Glyzyrrhiza 7"
Authors:   Institute of General and Experimental Biology, Kornopoltseva Tatiana V. , Botoeva Elena A. ,   Buryat State University
Published: March 2021

A promising direction for the search and development of new adaptogenic drugs of natural origin is the study of nonspecific drugs from the arsenal of Tibetan medicine, recommended for the weakened, as well as for elderly people, as a general tonic, "giving longevity and health." Based on the prescription recipe "Glyzyrhiza-7", in the indications for use: "with pain in the kidneys, lower back, limb stiffness, sudden flushes of heat in different parts of the body" developed a dry plant extract from the roots Glyzyrrhiza uralensis Fisch, wood Caragana jubata (Pall.) Poir, roots Polygonatum odoratum (Mill) Druse, roots Polygonatum humile Fisch ex Maxim, bulbs Orchis mascula L., roots Rheum rhabarbarum L., roots Rubia tinctorum L. HPLC in the dry extract revealed the presence of ten marker components whose raw material source is Rheum (deoxy-raponticin, raponticin, rapontigenin, caffeic acid), Glyzyrrhiza (likuritin, glycyrrhizinic acid), Rubia (alizarin, purpurine, ruberitrinic acid, lucidin-primverozid) The dominant compounds of the licorice 7 extract are anthraquinones, the content of which was 7.65 mg/g; stelben content 6.13 mg/g; terpenes (glycyrrhizic acid) 1.67 mg/g, flavonoids and phenol carboxylic acids account for 2.07 and 1.12 mg/g, respectively. A technique has been developed for the quantitative determination of the content of glycyrrhizic acid in the dry extract (16.5%), which can be used to standardize this object.
Die Open-Access-Transformation als gemeinschaftliches Ziel
Authors: Helmkamp Kerstin
Published: March 2021

In ihrem Beitrag führt Kerstin Helmkamp in die Themenfelder der der ersten Open-Access-Roadshow Schleswig-Holsteins, die vom 11. bis 14. November 2019 in Kiel, Flensburg und Lübeck
stattgefunden hat, ein. Sie benennt die Herausforderungen der Open-Access-Transformation, stellt das Vorgehen in Schleswig-Holstein der vergangenen Jahre dar und benennt zukünftige Perspektiven.
Grußwort des Ministeriums für Bildung, Wissenschaft und Kultur des Landes Schleswig-Holstein
Authors: Grundei Oliver
Published: March 2021

In seinem Grußwort für das Ministerium für Bildung, Wissenschaft und Kultur des Landes Schleswig-Holstein beschreibt Oliver Grundei die politisch gesetzten Rahmenbedingungen der Open-Access-Transformation.
Translation studies in German as a foreign language (GFL) in Africa (Nigera): A tool against "waiting room dangers"
Authors:   University of the Free State, Fakayode Omotayo
Published: March 2021

The importance of translation in Additional Language Learning (ALL) cannot be overemphasized, as there exist of late various studies in support of this fact. Extant studies on this issue have dealt with various aspects of translation in language learning in Europe, Canada, Australia and in the United States, although studies in this regard are not limited to these geographical boundaries. Moreover, these studies do not only focus on the impact of translation on ALL but also on various other issues like plurilingualism as it relates to translation in ALL. However, research in this regard are relatively scarce in the African context. As such, many studies done in Africa, particularly in Nigeria, are limited in scope when it comes to the relevance of translation in learning German as a Foreign Language (GFL). This study therefore seeks to shed more light on how translation studies in GFL lessons in Nigeria could be useful as a tool against “waiting room dangers”. In this sense, I mean the (in)security issues encountered in places of temporary localisations.
First Observed Interaction of the Circumstellar Envelope of an S-star with the Environment of Sgr A*
Authors: Peißker Florian , Ali Basel , Zajaček Michal , Eckart Andreas , Hosseini S. Elaheh , Karas Vladimír , Clénet Yann , Sabha Nadeen B. , Labadie Lucas , Subroweit Matthias
Published: March 2021

Several publications highlight the importance of the observations of bow shocks to learn more about the surrounding interstellar medium and radiation field. We revisit the most prominent dusty and gaseous bow shock source, X7, close to the supermassive black hole, Sgr A*, using multiwavelength analysis. For the purpose of this study, we use Spectrograph for Integral Field Observations in the Near Infrared (SINFONI) (H+K-band) and NACO L′- and M′-band) data sets between 2002 and 2018 with additional COMIC/ADONIS+RASOIR (L′-band)
7

7

COME-ON-PLUS Infrared Camera/Adaptive Optics Near Infrared System + Renouveau de l’Analyseur de Surface d’Onde InfraRouge

data of 1999. By analyzing the line maps of SINFONI, we identify a velocity of ∼200 km s−1 from the tip to the tail. Furthermore, a combination of the multiwavelength data of NACO and SINFONI in the H-, K-, L′-, and M′-bands results in a two-component blackbody fit that implies that X7 is a dust-enshrouded stellar object. The observed ongoing elongation and orientation of X7 in the Brγ line maps and the NACO L′-band continuum indicate a wind arising at the position of Sgr A* or at the IRS16 complex. Observations after 2010 show that the dust and the gas shell seems to be decoupled in the projection from its stellar source S50. The data also implies that the tail of X7 thermally heats up due to the presence of S50. The gas emission at the tip is excited because of the related forward scattering (Mie scattering), which will continue to influence the shape of X7 in the near future. In addition, we find excited [Fe iii] lines, which together with the recently analyzed dusty sources and the Brγ-bar underline the uniqueness of this source.
Spectroscopic and Photometric Redshift Estimation by Neural Networks for the China Space Station Optical Survey (CSS-OS)
Authors: Zhou Xingchen , Gong Yan , Meng Xian-Min , Zhang Xin , Cao Ye , Chen Xuelei , Amaro Valeria , Fan Zuhui , Fu Liping
Published: March 2021

The estimation of spectroscopic and photometric redshifts (spec-z and photo-z) is crucial for future cosmological surveys. It can directly affect several powerful measurements of the universe, such as weak lensing and galaxy clustering. In this work, we explore the accuracies of spec-z and photo-z that can be obtained by the China Space Station Optical Surveys, which is a next-generation space survey, using a neural network. The one-dimensional Convolutional Neural Networks and Multi-Layer Perceptron (MLP, the simplest form of an artificial neural network) are employed to derive spec-z and photo-z, respectively. The mock spectral and photometric data used for training and testing the networks are generated based on the COSMOS catalog. The networks have been trained with noisy data by creating Gaussian random realizations to reduce the statistical effects, resulting in a similar redshift accuracy for data with both high and low signal-to-noise ratios. The probability distribution functions of the predicted redshifts are also derived via Gaussian random realizations of the testing data, and then the best-fit redshifts and 1σ errors also can be obtained. We find that our networks can provide excellent redshift estimates with accuracies of ∼0.001 and 0.01 on spec-z and photo-z, respectively. Compared to existing photo-z codes, our MLP has a similar accuracy but is more efficient in the training process. The fractions of catastrophic redshifts or outliers can be dramatically suppressed compared to the ordinary template-fitting method. This indicates that the neural network method is feasible and powerful for spec-z and photo-z estimations in future cosmological surveys.
CRAFTS for Fast Radio Bursts: Extending the Dispersion–Fluence Relation with New FRBs Detected by FAST
Authors: Niu Chen-Hui , Li Di , Luo Rui , Wang Wei-Yang , Yao Jumei , Zhang Bing , Zhu Wei-Wei , Wang Pei , Ye Haoyang , Zhang Yong-Kun , Niu Jia-rui , Tang Ning-yu , Duan Ran , Krco Marko , Dai Shi , Feng Yi , Miao Chenchen , Pan Zhichen , Qian Lei , Xue Mengyao , Yuan Mao , Yue Youling , Zhang Lei , Zhang Xinxin
Published: March 2021

We report three new FRBs discovered by the Five-hundred-meter Aperture Spherical radio Telescope (FAST), namely FRB 181017.J0036+11, FRB 181118, and FRB 181130, through the Commensal Radio Astronomy FAST Survey (CRAFTS). Together with FRB 181123, which was reported earlier, all four FAST-discovered FRBs share the same characteristics of low fluence (≤0.2 Jy ms) and high dispersion measure (DM, >1000 pc cm−3), consistent with the anticorrelation between DM and fluence of the entire FRB population. FRB 181118 and FRB 181130 exhibit band-limited features. FRB 181130 is prominently scattered (τ

s
≃ 8 ms) at 1.25 GHz. FRB 181017.J0036+11 has full-bandwidth emission with a fluence of 0.042 Jy ms, which is one of the faintest FRB sources detected so far. CRAFTS has started to build a new sample of FRBs that fills the region for more distant and fainter FRBs in the fluence–DME diagram, previously out of reach of other surveys. The implied all-sky event rate of FRBs is

1.24

−
0.90

+
1.94

×

10

5

sky−1 day−1 at the 95% confidence interval above 0.0146 Jy ms. We also demonstrate here that the probability density function of CRAFTS FRB detections is sensitive to the assumed intrinsic FRB luminosity function and cosmological evolution, which may be further constrained with more discoveries.
Coverage versus Confidence
Authors: Cook Peyton
Published: March 2021

This article is intended to help students understand the concept of a coverage probability involving confidence intervals. Mathematica is used as a language for describing an algorithm to compute the coverage probability for a simple confidence interval based on the binomial distribution. Then, higher-level functions are used to compute probabilities of expressions in order to obtain coverage probabilities. Several examples are presented: two confidence intervals for a population proportion based on the binomial distribution, an asymptotic confidence interval for the mean of the Poisson distribution, and an asymptotic confidence interval for a population proportion based on the negative binomial distribution.
The composition of fish species shows little seasonal variation in eDNA based survey at urban streams
Authors: Kang Yujin , Song Youngkeun
Published: March 2021

Biodiversity in the city is becoming important as cities are expanded. Thus, the cost-efficient and accurate methodology for the investigation of biota has been consistently demanded. Environmental DNA (eDNA) is one of the emerging tools to supplement traditional survey methods and improve the efficiency of the survey. Summer (from July to August) is the monsoon period in South Korea, so it is excluded from the survey period because of the unstable aquatic environment, the safety of investigators, and accessibility to study sites. On the other hand, spring and autumn (from March to October), excluding the flooding season, are considered suitable times for the survey. This study compared fish composition and distribution by season in summer and autumn using eDNA to identify the change of fish composition and determine the suitability of the eDNA survey for seasonal fish investigation at urban streams. The number of eDNA reads, fish composition, and their community structures was changed according to the season. The total eDNA read was 2.3 times more in October (1,419,062) than in July (3,202,626). In addition, compared to the summer when 13 families and 33 species were detected, 13 families and 35 species were detected in fall. A total of 13 families and 32 species were detected in common. Three species (Acheilognathus rhombeus, Tachysurus fulvidraco, Pungtungia herzi) more were identified, and one (Hemiculter leucisculus) was not detected in October. The result described that the distribution rate of fish species has changed in the study area. Especially, the appearance of invasive species increased from 14 to 20 sites and the habitat range of invasive species was extended in October than July. However, there was no significant difference in richness and diversity except for abundance among fish community structures. This result revealed that eDNA survey methods can identify changes in the composition and distribution of fish by season. Furthermore, based on the little fluctuation in the species appearing according to the season, the study suggested that eDNA can be used to complement traditional survey methods by conducting fish surveys regardless of the survey period.
An eDNA-based method for monitoring a salmonid infectious disease: Development and application
Authors: Duval Eloïse , Blanchet Simon , Quéméré Erwan , Jacquin Lisa , Veyssière Charlotte , Lautraite Armand , Garmendia Laurent , Yotte Allan , Parthuisot Nathalie , Côte Jessica , Loot Géraldine
Published: March 2021

In the current context of global change, freshwater species are increasingly exposed to emerging infectious diseases (Okamura and Feist 2011). As an example, the Proliferative Kidney Disease (PKD) has emerged in salmonid fish during the last two decades, both in Europe and North America, causing important losses in aquaculture and worrying declines of several wild salmonid populations (Sudhagar et al. 2019). It is caused by Tetracapsuloides bryosalmonae, a myxozoan parasite with a complex life cycle involving two hosts: salmonids (intermediate host) and bryozoans (primary host). As PKD development strongly depends upon water temperature and quality, it is expected that global change could lead to more outbreaks (Okamura et al. 2011). Current monitoring of fish parasite load and infection status relies on histological observation or T. bryosalmonae DNA amplification out of kidney samples, involving fish euthanasia, and thus relatively small sample sizes when inferring infection prevalence. As large-scale screening of this parasite infections are required to better understand PKD dynamics, we have developed a non-lethal method for T. bryosalmonae detection in fish host based on the biological fact that T. bryosalmonae spores can be excreted from infected fish into the water through urine (Hedrick et al. 2004). This novel approach based on the detection of T. bryosalmonae DNA in fish urine was developed on wild brown trout (Salmo trutta), a species known to be an intermediate host of T. bryosalmonae and for releasing infective spores (only towards bryozoan host) through urine (Okamura et al. 2011). Applying this method, we have been able to map wild brown trout infection prevalence across 50 sites at the foothill of French Pyrenees and to identify the main environmental drivers of this disease.
Beyond fish eDNA metabarcoding: Field replicates disproportionately improve the detection of stream-associated vertebrate species
Authors: Macher Till-Hendrik , Schütz Robin , Arle Jens , Beermann Arne , Koschorreck Jan , Leese Florian
Published: March 2021

Fast, reliable, and comprehensive biodiversity monitoring data are needed for environmental decision making and management. Recent work on fish environmental DNA (eDNA) metabarcoding shows that aquatic diversity can be captured fast, reliably, and non-invasively at moderate costs. Because freshwater ecosystems act as sinks in the landscape, they also collect traces of terrestrial species via surface runoff or when specimens come into direct contact with water (e.g., for drinking purposes). Thus, fish eDNA metabarcoding data can provide information on fish but also on other, even terrestrial vertebrate species that live in riparian habitats. This data become available and may offer a much more comprehensive approach for assessing vertebrate diversity at no additional costs. Studies on how the sampling strategy affects species detection especially of stream-associated communities, however, are scarce. We therefore performed an analysis on the effects of biological replication on both fish as well as (semi-)terrestrial species detection. Along a 2-km stretch of the river Mulde (Germany), we collected 18 1L water samples and analyzed the relation of detected species richness and quantity of biological replicates taken. We detected 58 vertebrate species, of which 25 were fish and lamprey, 18 mammals, and 15 birds, which account for 50%, 24% and 7% of all native species to the German federal state of Saxony-Anhalt. However, while increasing the number of biological replicates resulted in only 25 % more detected fish and lamprey species, mammal and bird species richness increased disproportionately by 69 % and 84 %, respectively. Contrary, PCR replicates showed little stochasticity. We thus emphasize to increase the number of biological replicates when the aim is to improve general species detections. This holds especially true, when the focus is on rare aquatic taxa or on (semi-)terrestrial species, the so-called ‘bycatch’. As a clear advantage, this information can be obtained without any additional sampling or laboratory effort when the sampling strategy regarding biological replication is chosen carefully. With the consideration of frequent eDNA metabarcoding as part of national biomonitoring programs, the additional information provided by the bycatch can be used to further investigate the state of the environment and its biodiversity on a much broader scale.
Evaluating the ecological status of mountain streams from a megacity (Mexico City) with diatoms: development and implementation of morphological and eDNA-based methods
Authors: Salinas Camarillo Victor Hugo , Carmona-Jiménez Javier , Abarca Nelida , Zimmermann Jonas , Mora Demetrio
Published: March 2021

In central Mexico, streams flow from mountain protected areas at high elevations, i.e. abive 2,400 m a.s.l., being exposed to different human activities along their course. The main human threats of these aquatic ecosystems include the presence of hydraulic infrastructure (e.g. canals and dams), the construction of bridges or roads, fish farming, agriculture, livestock, unregulated tourism or irregular human settlements, resulting in nutrient enrichment, organic pollution and low hydromorphological quality. Diatoms, widely recognized as valuable indicators of environmental conditions, offer a unique opportunity to assess the ecological quality of the streams in and around this megacity, as they respond directly and sensitively to specific physical, chemical and biological changes. To conduct ecological assessment, the composition of diatom communities has traditionally been characterized by morphological methods and most recently by environmental DNA (eDNA) metabarcoding. However, the diatom communities of only a few of these streams in the Basin of Mexico have been characterized morphologically at low resolution light microscopy (LM), and no eDNA-based studies have been conducted yet. In this context, our aims are to 1) implement morphological and eDNA metabarcoding methods for the identification and quantification of epilithic diatoms and to assess their seasonal and spatial response in three peri-urban mountain streams from the Basin of Mexico, and 2) develop a metric to assess and monitor the ecological quality in these streams. For this purpose, we characterized a) the physical, environmental and hydro-morphological conditions related to ecosystem quality, and b) the diatom diversity and community structure of nine sites per stream (27 sites). The sites include headwaters, as well as middle and lower reaches with different land-use and anthropologic pressures, and were investigated over three hydrological periods, i.e. post-rainy, dry-cold and dry-warm. Species identification was first conducted under LM and scanning electron microscopy (SEM), and is currently been done by eDNA metabarcoding of the V4 region of the 18S rRNA gene and a fragment of the chloroplast marker rbcL. Our study has resulted in 520 infrageneric taxa identified by LM and SEM, the largest diversity ever found for freshwater diatoms of Mexico. From this data, we found a clear environmental gradient from almost pristine conditions towards heavily polluted states as the streams enter the city. These changes are well reflected by variations in the diatom community structure, as revealed by the Diatom Ecological Quality Index (DEQI), newly developed to evaluate the ecological quality of the streams in and around this megacity.
Assessment of the macroinvertebrate community of the Vjosa river through non-destructive DNA metabarcoding of preservative ethanol
Authors: Brasseur Marie , Vitecek Simon , Zizka Vera , Martini Jan , Wüthrich Remo , Birnstiel Emil , Wilfling Olivia , Singer Gabriel , Leese Florian
Published: March 2021

Streams and rivers represent hotspots of biodiversity in their natural state. This biodiversity is declining worldwide due to pollution, exploitation and hydromorphological degradation of these systems. One of the last big, natural rivers in Europe is the Vjosa in the Balkan region. The catchment is characterized by natural flow dynamics, resulting in high habitat diversity and turnover, and hosts several sensitive and endemic species (e.g. Isoperla vjosae). DNA metabarcoding represents a promising approach to characterize this biodiversity but methodological drawbacks such as primer bias or incomplete reference databases limit the application, particularly in taxonomically underexplored regions.

Here, we assessed stream biodiversity with a focus on macrozoobenthic (MZB) taxa via a non-destructive, voucher-preserving DNA metabarcoding protocol. In this approach, ethanol used for preservation of multi-habitat samples in the field was used as DNA template, allowing to retain the integrity of the original sample and further comparison of molecular and morphological taxa lists. Samples were taken in spring and autumn 2018 at 48 sites allocated over the Vjosa catchment. The preservative ethanol was filtered through 0.43 um nitrocellulose membranes from which DNA was extracted. Subsequently, a 421 bp fragment of the COI gene was amplified with the primer pair BF2/BR2 and Illumina sequenced. After filtering for sequences with similarity to reference entries >85%, 4,123 OTUs were obtained, of which 921 were identified as MZB taxa. Dipterans and ephemeropterans were most abundant, followed by plecopterans. Some taxa (e.g. molluscs) were not identified due to a known primer bias and >7,000 OTUs could not be assigned above 85 % similarity. Using the here presented voucher-preserving approach allowed us to identify the pitfalls of DNA metabarcoding as tool for biodiversity assessment in taxonomically unexplored regions such as the Vjosa catchment. However, the comparison of specimen abundance data and molecular data showed the power of non-destructive fixative metabarcoding for detecting MZB communities with highly increased taxonomic resolution.
Dots on Earth with huge genetic diversity: relict flatworm Crenobia alpinain the alpine lakes of Tatra Mountains
Authors: Tuhrinová Kornélia , Bozáňová Jana , Čiampor Jr Fedor , Čiamporová-Zaťovičová Zuzana
Published: March 2021

Mountain lakes, typical signs of an alpine landscape all around the world, were formed in the western part of the Carpathian Arch about 10,000-15,000 years ago, as remnants of the last continental glaciation. In the Tatra Mountains (Slovak-Polish border), more than 110 permanent lakes of glacial origin and many other small ponds exist. These (sub)alpine lakes, with their specific conditions, are very vulnerable and any change in their environment can largely impact their biodiversity, which is, despite long-term faunistic and ecological research, still not fully recognized. This shortcoming could be strongly improved by molecular approaches, which, even within population-genetic studies, often reveal unsuspected cryptic lineages or potential new species.

Our long-term research is aimed at revealing the genetic diversity of aquatic macroinvertebrate species in the alpine environment of the Western Carpathians. One of the currently studied species is the glacial relict flatworm taxon Crenobia alpina (Dana, 1766) (Tricladida: Planariidae), representing an element of the permanent aquatic fauna. Totally, 88 COI haplotypes of C. alpina forming at least five well-separated genetic lineages were identified within 348 individuals collected from 45 lakes and ponds in theTatra Mts. Based on tests of selective neutrality and population stability, C. alpina populations in the Tatra lakes did not overcome recent population expansions or contractions. It seems that great genetic differences between localities and their population stability could be a consequence of the presence of natural topographic barriers (ridges, mountain peaks) dividing the mountains into small and relatively isolated valleys. Moreover, other important factors that could play a significant role are the limited dispersal ability of the species as well as its ability to reproduce asexually. Our findings were also compared with all publicly available sequences (Barcoding of Life Datasystems - BOLD and GenBank) in order to shift our data from a local to a holistic view on C. alpina. Our resultssupport the assumption of Brändle et al. 2017 that C. alpina represents a complex of cryptic lineages or species in Europe. As part of the AquaBOL.sk initiative, the data obtained contribute to the knowledge of genetic variability and barcoding of aquatic organisms in Slovakia.
Gap analysis for DNA barcodes of aquatic macroinvertebrate species in the Southeast of Italy
Authors: Specchia Valeria , Zangaro Francesco , Tzafesta Eftychia , Pinna Maurizio
Published: March 2021

DNA metabarcoding for the identification of species and ecosystem biomonitoring is a promising innovative approach. The applicability of this tool is at first dependent on the coverage of the DNA sequence reference libraries. We performed a gap analysis of available DNA barcodes in the international databases using the aquatic macroinvertebrate species checklist of the Apulia region in southeast Italy. Our analyses show that 42% of the 1546 examined species do not have representative DNA barcodes in the reference libraries, indicating the importance of working toward their completeness and addressing this effort toward specific taxonomic groups in particular at local/regional level. The DNA-barcode coverage also varies among different taxonomic groups and aquatic ecosystem types in which a large number of species are rare. We also analyzed the DNA barcode reference libraries for the primer set used to barcode species. Only for 52% of the examined barcoded species were the primers reported, indicating the importance of uploading this information in the databases for a more extensive use of the DNA metabarcoding. We also highlighted the opportunity to develop combinations of primers useful at the regional level. We tested the application of the DNA barcoding single species to a lagoon ecosystem (the lagoon named “Aquatina di Frigole” in the Apulia region) which are richer in humic substances than other aquatic environments and in which DNA metabarcoding remains under explored.
Year-round monitoring and large-scale screening of native and invasive crayfishes in lotic systems
Authors: Chucholl Franziska , Epp Laura , Segelbacher Gernot
Published: March 2021

Freshwater crayfish are among the most threatened animal taxa in Central Europe. Effective conservation measures for endangered native and management of invasive alien crayfishes requires knowledge about distribution, monitoring of existing and early detection of newly established populations. eDNA has recently emerged as a promsing, highly sensitive, and non-invasive detection tool in this regard.

To evaluate eDNA as detection tool for freshwater crayfish, we developed a novel set of specific eDNA-assays for all native (Austropotamobius torrentium, Austropotamobius pallipes, Astacus astacus) and the most relevant invasive crayfish species (Pacifastacus leniusculus, Faxonius limosus, Faxonius immunis) in Central Europe. To ensure specificity each primer pair was tested in silico, in vitro, and in situ, including a total of 13 lotic and lentic waterbodies (Fig. 1). Moreover, we assessed the influence of spatio-temporal variables (distance to upstream population, season, and stream size) on eDNA detection in seven streams using two different detection methods (qualitative endpoint PCR and quantitative droplet digital PCR, ddPCR).

The newly developed eDNA assays successfully detected all crayfish species across different lotic and lentic habitats. Multiple linear mixed-effects analysis revealed a significant effect of distance and season on eDNA detection rate (endpoint PCR) and eDNA-concentration (ddPCR). Year-round detection was successful up to 7 km downstream of the source population, although detectability was lowest in winter. eDNA detection rate further decreased with increasing stream size. Finally, eDNA-concentration correlated positively with estimated upstream population size.

Overall, our study provides easily applicable eDNA assays for six crayfish species, enabling year-round detection, which represents a clear benefit over conventional methods. Due to its high sensitivity, eDNA detection is also suitable for the targeted search of as-yet unrecorded or newly emerging populations. Using quantitative ddPCR might further allow for a rough estimation of population size, provided that the identified spatio-temporal factors are accounted for. We therefore recommend implementing eDNA-detection as a complementary survey tool, particularly for a large-scale screening of data-deficient catchments or a year-round monitoring (Chucholl et al. 2021).
eDNA metabarcoding survey of fish communities in the Danube and its tributaries
Authors: Pont Didier , Meulenbroek Paul , Bammer Vinzenz , Dejean Tony , Eros Tibor , Hein Thomas , Jean Pauline , Lenhardt Mirjana , Nagel Christoffer , Pekárik Ladislav , Schabuss Michael , Stoeckle Bernhard , Weigand Alexander , Zornig Horst , Valentini Alice
Published: March 2021

In complement to the JDS4 traditional fish survey (TFS, mainly by electrofishing), a fish eDNA metabarcoding-based survey has been implemented within the framework of the monitoring program organized by DNAqua-Net and in collaboration with the INTERREG "MEASURES" program. Water samples were collected at 29 sites from the source to the mouth of the Danube River, and at 18 tributaries. Two water samples (mean volume 29 L per sample) were collected at each site and the water filtered in situ. Twelve PCR replicates were performed per sample using teleo primers.

In total, 80 taxa were detected, of which 19 corresponded mainly to farmed fish or food fish due to eDNA release in waste waters, most often downstream of large cities. Of the remaining 61 taxa, 50 taxa were identified at the species level, six taxa at the genus level, and five taxa at a higher taxonomic level. Concerning the Danube river itself, 69 and 57 species/markers were detected along the Danube River by TFS and eDNA surveys, respectively. 50 of these taxa were detected by both methods. Nine species were captured by TFS alone, mainly due to gaps in reference librairies and too colosely related barcodes. Eight species were only detected by eDNA. Except for the Salvelinus group, these were all benthic species, which are difficult to catch by electrofishing in large rivers (Acipenser ruthenus, Acipenser stellatus, Benthophilus sp., Romanogobio uranoscopus, Sabanejewia balcanica, Umbra krameri).

The species richness tended to increase from upstream to downstream and the relative number of DNA sequences per taxa showed a clear succession of species along the river. Barbatula barbatula, Cottus gobio, Hucho hucho, Lampletra planeri, Phoxinus phoxinus and Thymallus thymallus were restricted to the Upper Danube whereas A. ruthenus, Neogobius fluviatilis, S. balcanica and Scardinius erythrophtalmus were detected from Vienna to the Danube river mouth. Abramis brama, Alburnus alburnus, Cyprinus carpio, Silurus glanis, Sander spp, Zingel streber were detected all along the river course; Alosa spp. and Syngnathus abaster downstream from the Iron Gate; A. stellatus and U. krameri only on the most downstream site (Danube delta).

The calculation of a fish index, based on the common metrics used to intercalibrate national fish assessment methods on a European scale, classified most of the sites as being in moderate ecological status. Comparison of the indicative ecological status calculated using the same assessment method based on TSF and eDNA data at the common Danube sites showed a similar classification for six of the 13 sites and a difference of one class for the remaining seven sites.
Development of a set of oligonucleotides for the identification of the Sakhalin sturgeon Acipenser mikadoi Hilgendorf, 1892 by PCR
Authors: Turanov Sergei
Published: March 2021

Sturgeons (family Acipenseridae) are valuable commercial fish and aquaculture resources. The Acipenser genus includes about 20 species, 12 of which are recorded for the Russian territory. One of the rarest is the Sakhalin sturgeon A. mikadoi Hilgendorf, 1892. At present, its population size is significantly decreased and the species is close to extinction. Natural populations of the species have survived in the Tumnin River in Khabarovsk Region and the Viakhtu River in Sakhalin Region. Vihtu River in Sakhalin Region. Due to the small numbers of rare and endangered fish species, noninvasive approaches, including the use of DNA from the environment, appear to be the most suitable for monitoring their species and genetic diversity. The method is well established in this area and has been successfully tested for the monitoring of several species of rare and endangered sturgeons. This paper presents the results of the development of primers specific for the mitochondrial DNA of the species A. mikadoi.

A matrix of 15 sequences of complete mitochondrial genomes from 3 sturgeon species whose ranges may overlap - A. mikadoi (KX276658), A. dauricus (KJ402277) and A. schrenckii (MH973728- MH973734, KX276660, KX276659, KF150287, KC905169, KC820796) - was generated. A. medirostris (NC_028405), which is genetically closest to the Sakhalin sturgeon, was also included in the analysis. A sliding window algorithm was applied to the sequence matrix. A graph showing the distribution of divergence values along the mitochondrial genome of sturgeon was plotted based on the results of the analysis Fig. 1.

Based on matrices of individual genome fragments (16S rRNA, COI, ND1, and D-loop), a search for species-specific primers was performed in the DECIPHER program. A Primer-BLAST algorithm was run based on each pair of primers. Taxa Actinopterygii and Acipenseridae were used as references separately in different runs. In addition, primers were checked using local reference sequences of the complete mitochondrial genome of representatives of the genus Acipenser. The results of primer evaluation are summarized in the table Table 1. As a result, a combination of primer pairs developed on the basis of D-loop and COI fragments seems to be the most successful for error-free identification of Sakhalin sturgeon.