Ethephon (ETF) has been used in agriculture as an ethylene releaser type of plant growth regulator. The aim of this work was to determine the ecotoxicological effects of ETF on the survival and the antioxidant metabolism of the insects using a model organism Galleria mellonella L. 1758. A toxicity test was performed to determine the lethal doses of ETF on larvae. According to probit assay, the LD50 and LD99 values for force fed larvae were 344 and 419 µg/5 µl, respectively, 30 d after treatment. Analyses performed with 10 doses ≤LD50 at 24 and 48 h upon feeding larvae revealed that the malondialdehyde level increased at 300 and 330 µg/5 µl doses, whereas glutathione-S-transferase activity increased only with a 360 µg/5µl dose of ETF at 24 h. However, an increase in glutathione-S-transferase activity was evident at all ETF doses at 48 h. An increase in glutathione peroxidases activity was determined at 250, 300 and 330 µg/5µl at 24 and 48 h. All ETF doses caused an important increase in catalase activity at 24 h but remained unchanged at 48 h. Superoxide dismutase activity also elevated at doses >250 µg/5µl at 24 h when compared to the control. Same changes in superoxide dismutase activity were also observed at all doses of ETF except for 360 µg/5µl at 48 h. These results showed that ETF induced oxidative stress resulted in toxic effects that affected on the survival of model organism G. mellonella. ÖzetEtilen salınımına neden olan Etefon (ETF), bir bitki büyüme düzenleyicisi olarak tarımda kullanılmaktadır. Bu çalışmada bir model organizma olan Galleria mellonella L. 1758 (Lepidoptera: Pyralidae) türü kullanılarak, ETF'nin böceklerin antioksidan metabolizması ve canlılığı üzerindeki ekotoksikolojik etkilerinin araştırılması amaçlanmıştır. Larval dönemde ETF'nin letal dozunun belirlemesi amacıyla toksitite testi yapılmıştır. Larvalara zorla besleme (ağızdan besleme) yöntemi ile uygulanan ETF dozlarına göre, 30 günlük süreç içinde belirlenen LD50 ve LD99 değerleri sırasıyla 344 ve 419 µg/5µl olarak belirlenmiştir. LD50 ve daha düşük ETF dozlarıyla yapılan toksikolojik analizlerde ise iki zaman dilimi (24. ve 48. saat) tercih edilmiştir. 24. saatte, larval hemolenfteki malondialdehit seviyesi, 300 ve 330 µg/5µl ETF dozlarında artarken, glutatyon-S-transferaz aktivitesi sadece 360 µg/5µ'lik dozda yükselmiştir. Ancak 48. saatte kontrol ve tüm dozlarda glutatyon-S-transferaz aktivitesi yükselmiştir. Glutatyon peroksidaz aktivitesi ise hem 24. hem de 48. saatte, 330 ve 360 µg/5µl ETF dozlarında artmıştır. Tüm ETF dozları 24. saatte katalaz aktivitesinde artışa neden olurken, bu artış 48. saatte de aynı kalmıştır. Süperoksit dismutaz aktivitesi ise 24. saatte, 250 µg/5µl ve daha yüksek dozlarda yükselmiştir. 48. saatteki süperoksit dismutaz aktivitesinde de benzer değişimler meydana gelirken, 360 µg/5µl dozunda azalma belirlenmiştir. Bu sonuçlar, ETF'nin oksidatif stresi teşvik etmesi sonucunda model organizma G. mellonella'nın canlılığı üzerinde toksik etkisi olduğunu göstermiştir.
SARS-CoV-2 in vitro transcribed RNA reference materials (RM), UME RM 2019 and UME RM 2020, were produced by Scientific and Technological Research Council of Turkey (TUBITAK), National Metrology Institute (UME), to be used as a quality control material for SARS-CoV-2 measurements, in liquid-frozen and lyophilized forms, respectively. These RNA RMs include ten internationally recommended SARS-CoV-2 target gene fragments (Pasteur-RdRp-IP2, Pasteur-RdRp-IP4, Charite-E, Charite-RdRp, CDC-N1, CDC-N2, China CDC-ORF1ab, China CDC-N, Hong Kong-ORF1b, and Hong Kong-N) for virus detection and one human gene fragment (RNase P) as an internal control. Two different platforms, RT-qPCR and RT-ddPCR, were used to characterize UME RM 2019 (UME RM 2020 was only characterized with RT-qPCR). The homogeneity studies were evaluated by RT-qPCR. According to these results, it has been shown that both reference materials are homogeneous for intended use. Short-term studies were also conducted similarly for mimicking transport conditions and UME RM 2020, which is produced in lyophilized form, unlike other reference materials available in the market, provides convenience for users by ensuring that the reference material remains stable for 17 days even at 45°C temperature. The lyophilized formulation of the reference material had greater stability which would allow it to be shipped without cooling items. The development of such RNA reference materials provides quality control for existing and newly designed RNA-based virus detection tests and it helps the prevention and control of epidemics.
Background Tabanus bromius (Diptera: Tabanidae) is one of the most notable Tabanidae species of veterinary and medical importance distributed throughout the Palearctic region. In this study, we investigate the genetic diversity and the phylogeographic structure of T. bromius sampled from Turkey, Croatia, and Iran. Methods For this purpose, a 686-base-pair (bp) fragment of mitochondrial DNA cytochrome oxidase I gene (COI) and 1339 bp of the nuclear DNA internal transcribed spacer (ITS) were sequenced from 247 individuals representing 15 populations. Results The sequences generated 169 COI haplotypes and 90 ITS alleles. A higher haplotype/allele diversity (h = 0.9909 for the COI gene and Ad = 0.8193 for the ITS region) compared to a low nucleotide diversity (π = 0.020605 for COI gene and π = 0.013667 for the ITS region), present for a high number of singleton and private haplotypes/alleles imply population expansion in the past. The results of phylogenetic analysis led to the uncovering of geographically significant groupings of lineages with regard to the entrance of the species into Anatolia and the location of major geographic barriers. According to current data, the species appears to have entered Turkey from Caucasia and Iran. A molecular clock applied to the COI data suggests that T. bromius diverged from the outgroup species nearly 8.83 million years ago, around the end of the Miocene era. Conclusions The results of this study indicate remarkable genetic diversity across the studied range of the species. High haplotype/allele versus low nucleotide diversity and demographic analyses implied that the T. bromius populations have undergone a series of expansions and retreats in the past. Our current findings suggest that T. bromius split from outgroups around the Late Miocene. Subsequent diversification events during the climatic and environmental fluctuation times of the Late Pliocene and Early Pleistocene periods also significantly influenced the species, resulting in the formation of some major genetic lineages. The phylogenetic analyses indicate that T. bromius most likely entered Turkey from the Caucasus region and Iran. Graphical Abstract
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