Skin swabs with FTA® cards as a dry storage source for amphibian DNA

Amphibians are the most endangered group of vertebrates, and conservation measures increasingly rely on information drawn from genetic markers. The present study explores skin swabs with Whatman FTA® cards as a method to retrieve PCR-amplifiable amphibian DNA. Swabs from ten adult great crested newts (Triturus cristatus) were used to compare FTA® card-based protocols with tissue sampling based on toe clips. PCR success rates were measured for seven microsatellite markers and one mtDNA marker (ND4) after 6 months of sample storage. We demonstrate that the merging of eight FTA® card punches from Qiagen-based DNA extraction always led to successful amplifications in at least one replicate, at an overall PCR success rate of 78%. The newly established protocol has the potential for wide application to future DNA-based amphibian studies

Biologische Kontrolle von Eulenraupen im Kohl mittels Baculoviren

Baculoviren sind als hoch selektive biologische Kontrollagenzien für Kohleulenraupen (Mamestra brassicae) seit längerem bekannt und ihre gute Wirksamkeit wurde schon in verschiedenen Gewächshaus- und Freilandversuchen nachgewiesen. Um die Kontrollsituation von Kohleulenraupen im ökologischen Gemüseanbau insbesondere bei niedrigen Temperaturen zu verbessern, wurden die eulenpathogenen Mamestra brassicae Nukelopolyhedrovirus (MbMNPV) und Mamestra configurata Nukleopolyhedrovirus (MacoNPV-A) als mögliche Kontrollagentien geprüft. Es konnte gezeigt werden, dass auch MacoNPV-A für M.brassicae pathogen ist. In vergleichenden Bioassays bei 24°C zeigten MbMNPV und MacoNPV-A keinen signifikanten Unterschied hinsichtlich ihrer biologischen Aktivität gegenüber M.brassicae. In weiteren Bioassays wurden die konzentrationsabhängige Mortalität von M.brassicae im 2. Larvenstadium sowie die gefressene Blattfläche bei 24, 20, 16 und 12°C bestimmt. Im Vergleich zu 24 und 20°C waren die mittleren Letalkonzentrationen bei 16 und 12°C 100 bzw. 1000fach höher. Für eine mittlere Reduktion der konsumierten Blattfläche waren hingegen bei Temperaturen unter 20°C 10 bis 100fach höhere Konzentrationen notwendig. Durch die Neuformulierung von MbMNPV (Probis GmbH, Wiernsheim) konnte eine Wirkungssteigerung im Vergleich zu der nicht formulierten Virussuspension im Bioassay erzielt werden. In einem Freilandtest mit künstlicher Infestation in Kohlrabi konnte eine weitere Steigerung des Wirkungsgrades durch die Kombination mit dem Bacillus thuringiensis Präparat XenTari® im Vergleich zur Einzelapplikation der beiden Präparate erreicht werden. Eine breite Freilandtestung der Baculoviren-Präparaten gegen Kohlraupen bei kühler Witterung als Einzelpräparat und in Kombination mit Bacillus thuringiensis Präparaten sowie neuester Applikationstechnik für Ober- und Unterblattbehandlung wird empfohlen

Effekten av olika koncentrationer av Rotstop R och Rotstop S och ofullständig täckning av Rotstop S på sporinfektioner av rotticka på granstubbar

The effectiveness of treatment with two Phlebiopsis gigantea based preparations (Rotstop® and RotstopS) in different concentrations against the root and butt rot causing fungus Heterobasidion annosum s.l. on Norway spruce (Picea abies) thinning stumps in southern Sweden was compared. The trees were cut on three sites during the summer 2004 and 285 stumps were treated manually with 100% cover with two different amounts of spores in solution corresponding to approximately 5×106 spores/l and 10×106 spores/l. 31 stumps received mechanical part cover with the highest spore concentrations of Phlebiopsis gigantea, i.e. approximately 20×106 spores/l. Mechanical treatment was assessed for satisfactory treatment effect. Three months later, samples were collected and analyzed. There was a significant reduction in frequency and relative areas of Heterobasidion spp. infections on stumps with manual application of control agents compared to untreated stumps. On average 10 to 23% of stumps subjected to manual treatments were infected compared to 52% for the untreated stumps. However, none of the concentration solutions of Rotstop® and RotstopS differed from the others in reduction of Heterobasidion spp. infections. Mechanical treatment failed to control the Heterobasidion spp. infections, but there was a tendency for the 85.1-95.0% cover class to provide better result than the others. Thus, despite of the incomplete control of the pathogen, stump treatment with different concentrations of Phlebiopsis gigantea oidia in suspensions provided equal effects at the present spore loads of Heterobasidion spp.Effektiviteten av behandling med två pergamentsvampsbaserade (Phlebiopsis gigantea) lösningar (Rotstop® och RotstopS) i olika koncentrationer mot sporinfektioner av rotticka (Heterobasidion spp.) på gallringsstubbar av gran (Picea abies) i södra Sverige jämfördes. Träd avverkades på tre olika lokaler under sommaren 2004 och sammanlagt 285 stubbar behandlades manuellt med en hundraprocentig täckning i två olika koncentrationer motsvarande ungefär 5×106 sporer/l respektive 10×106 sporer/l lösning. 31 stubbar på en av lokalerna behandlades maskinellt med en ofullständig täckning men med den högsta koncentrationen av pergamentsvamp (RotstopS), ca 20×106 sporer/l lösning. Effekten av den mekaniska behandlingens partiella täckning jämfördes med den manuella fullständiga. Tre månader efter behandlingen samlades prover in för analys med avseende på förekomsten av rotticka. Alla manuella behandlingar gav en signifikant reducering av frekvensen av och relativa arean av rottickeinfektioner jämfört med obehandlade stubbar. I genomsnitt var 10 till 23 % av de manuellt behandlade stubbarna infekterade jämfört med 52 % av de obehandlade. Ingen av de manuella applikationerna skiljde sig dock från varandra. Den mekaniska behandlingen gav inte ett tillfredsställande resultat men det fanns en tendens till att täckningsgradsklass 85,1- 95,0 % fungerade bättre än de andra. Således, trots att ingen behandling gav en 100 % -ig minskning av antalet rottickeinfektioner, verkar olika koncentrationer av Rotstop® och RotstopS ge ett likvärdigt resultat under de förhållanden i sportryck som rådde vid behandlingen

Untersuchungen zur Populationsgenetik der Minderempfindlichkeit des Apfelwicklers gegenüber Cydia pomonella Granulovirus (CpGV)

The Codling moth granulovirus (Cydia pomonella granulovirus, CpGV, Baculoviridae) is one of the most important bio control agents of the codling moth in apple production. Since 2003, codling moth populations have been observed in Germany and France, which show an up to thousand fold decreased susceptibility to CpGV. A spread of this phenomenon is a severe threat to the efficient control of the codling moth, particularly in organic farming. In order to prevent this development, investigations on the popula-tion genetics of codling moth populations in Germany were initiated to assess the baseline susceptibilities of selected populations. Furthermore, the genetic and biologi-cal background of resistance of the codling moth to CpGV are being elucidated by crossing susceptible and low susceptible codling moth populations. These investiga-tions will help to develop new control strategies or to restore high susceptibility to-wards CpGV. Mapping of traits involved in resistance will be performed. Involved loci will be identi-fied with the help of amplified fragment length polymorphism (AFLP). Loci coupled with susceptibility can help to elucidate resistance mechanisms. Analysis of comple-mentary DNA amplified fragment length polymorphism (cDNA-AFLP) will be per-formed to display differences in expression rate of particular genes. If there are differ-ences between sensitive and non-sensitive strains, the genes will be isolated and sequenced. Putative sequence homologies give the direction of the functional sense of the mentioned gene and further conclusion of the mechanisms of the susceptibility of CpGV

Kinematics and hydrodynamics of spinning particles

In the first part (Sections 1 and 2) of this paper --starting from the Pauli current, in the ordinary tensorial language-- we obtain the decomposition of the non-relativistic field velocity into two orthogonal parts: (i) the "classical part, that is, the 3-velocity w = p/m OF the center-of-mass (CM), and (ii) the so-called "quantum" part, that is, the 3-velocity V of the motion IN the CM frame (namely, the internal "spin motion" or zitterbewegung). By inserting such a complete, composite expression of the velocity into the kinetic energy term of the non-relativistic classical (i.e., newtonian) lagrangian, we straightforwardly get the appearance of the so-called "quantum potential" associated, as it is known, with the Madelung fluid. This result carries further evidence that the quantum behaviour of micro-systems can be adirect consequence of the fundamental existence of spin. In the second part (Sections 3 and 4), we fix our attention on the total 3-velocity v = w + V, it being now necessary to pass to relativistic (classical) physics; and we show that the proper time entering the definition of the four-velocity v^mu for spinning particles has to be the proper time tau of the CM frame. Inserting the correct Lorentz factor into the definition of v^mu leads to completely new kinematical properties for v_mu v^mu. The important constraint p_mu v^mu = m, identically true for scalar particles, but just assumed a priori in all previous spinning particle theories, is herein derived in a self-consistent way.Comment: LaTeX file; needs kapproc.st

Buprenorphine-Naloxone for Opioid Use Disorder: Reduction in Mortality and Increased Remission

Introduction: As fentanyl has become more readily available, opioid-related morbidity and mortality in the United States has increased dramatically. Preliminary studies suggest that high-affinity, partial mu-opioid receptor agonists such as the combination product buprenorphine-naloxone may reduce mortality from overdose and promote remission. With the escalating prevalence of opioid use disorder (OUD), it is essential to evaluate the effectiveness of opioid agonists like buprenorphine-naloxone. This study examines mortality and remission rates for OUD patients prescribed buprenorphine-naloxone to determine the efficacy of this treatment toward these outcomes. Methods: We carried out a retrospective analysis using the US Collaborative Network database in TriNetX, examining de-identified medical records from nearly 92 million patients across 56 healthcare organizations. The study spanned the years from January 1, 2017–May 13, 2022. Cohort 1 included OUD patients who began buprenorphine-naloxone treatment within one-year post-diagnosis, while Cohort 2, the control group, consisted of OUD patients who were not administered buprenorphine. The study measured mortality and remission rates within a year of the index event, incorporating propensity score matching for age, gender, and race/ethnicity. Results: Prior to propensity matching, we identified a total of 221,967 patients with OUD. Following exclusions, 61,656 patients treated with buprenorphine-naloxone showed 34% fewer deaths within one year of diagnosis compared to 159,061 patients who did not receive buprenorphine (2.6% vs 4.0%; relative risk [RR] 0.661; 95% confidence interval [CI] 0.627–0.698; P < 0.001). The remission rate was approximately 1.9 times higher in the buprenorphine-naloxone group compared to the control group (18.8% vs 10.1%; RR 1.862; 95% CI 1.812–1.914; P < 0.001). After propensity matching, the effect on mortality decreased but remained statistically significant (2.6% vs 3.0%; RR 0.868; 95% CI 0.813–0.927; P < 0.001) and the remission rate remained consistent (18.8% vs 10.4%; RR 1.812; 95% CI 1.750–1.876; P < 0.001). Number needed to treat for benefit was 249 for death and 12 for remission. Conclusion: Buprenorphine-naloxone was associated with significantly reduced mortality and increased remission rates for patients with opioid use disorder and should be used as a primary treatment. The recognition and implementation of treatment options like buprenorphine-naloxone is vital in alleviating the impact of OUD

Elucidating the genomic history of commercially used Bacillus thuringiensis subsp. tenebrionis strain NB176

Bacillus thuringiensis subsp. tenebrionis (Btt) produces a coleopteran-specific crystal protoxin protein (Cry3Aa δ-endotoxin). After its discovery in 1982, the strain NB125 (DSM 5526) was eventually registered in 1990 to control the Colorado potato beetle (Leptinotarsa decemlineata). Gamma-irradiation of NB125 resulted in strain NB176-1 (DSM 5480) that exhibited higher cry3Aa production and became the active ingredient of the plant protection product Novodor® FC. Here, we report a comparative genome analysis of the parental strain NB125, its derivative NB176-1 and the current commercial production strain NB176. The entire genome sequences of the parental and derivative strains were deciphered by a hybrid de novo approach using short (Illumina) and long (Nanopore) read sequencing techniques. Genome assembly revealed a chromosome of 5.4 to 5.6 Mbp and six plasmids with a size range from 14.9 to 250.5 kbp for each strain. The major differences among the original NB125 and the derivative strains NB176-1 and NB176 were an additional copy of the cry3Aa gene, which translocated to another plasmid as well as a chromosomal deletion (~ 178 kbp) in NB176. The assembled genome sequences were further analyzed in silico for the presence of virulence and antimicrobial resistance (AMR) genes