Gentamicin is an aminoglycoside antibiotic effective against aerobic gram-negative bacteria and is also used in veterinary medicine, particularly in the swine and bovine industries. However, no gentamicin product is currently approved for treating equine diseases in Korea. The present study aims to examine the time-dependent residue of gentamicin in horses after intravenous injection (IV) via jugular vein. The test product was injected at 6.6 mg/kg BW via jugular vein in nine horses. Blood was collected from the horse's jugular vein at 15 minutes, 30 minutes, 1, 4, 8, 12, 24 and 48 hours after injection. To purify the gentamicin in serum, 100μL of 20 mM HFBA in DW, 100 μL of 30% trichloroacetic acid and 300 μL of 20 mM heptafluorobutyric acid (HFBA) in acetonitrile (ACN) were added to 500 μL of serum and supernatant was applied to LC-MS/MS after centrifugation. LC-MS/MS-8050 analyzed the level of gentamicin in serum with Electrospray ionization (ESI) and multiple reaction monitoring (MRM) positive mode. Gentamicin C1 was 478 m/z and product ions were 322, 157 m/z. Precursor ion of Gentamicin C1a was 450 m/z and product ions were 322, 160 m/z. Precursor ion of Gentamicin C2 and C2a was 464 m/z and product ions were 322, 160 m/z. The LC column was a C18 and mobile phase composed of 20 mM HFBA in 5% ACN and 20 mM HFBA in 50% ACN. The amount of gentamicin was calculated by adding four components of gentamicin (C1, C1a, C2 and C2a). The pharmacokinetic parameters of gentamicin were calculated by the WinNonlin program. The Cmax of gentamicin in horse serum was 93 ± 17 μg/kg and the Tmax was 0.25 ± 0 hours. The T1/2 was 6.41 ± 2.32 hours and the CLt was 0.05 ± 0.01L/hr/kg. The Vd was shown as 0.44 ± 0.13 L/kg and the MRT was 1.98 ± 0.55 hours. In conclusion, our data provides useful pharmacokinetic parameters for gentamicin in horses following IV injection.
In this study, an simultaneous LC-MS/MS multi-residue analytical method was developed and validated for the residues of six neonicotinoid insecticides (acetamiprid, clothianidin, dinotefuran, imidacloprid, thiacloprid, and thiamethoxam) in honey. Sample preparation included a combination of QuEChERS extraction kit and liquid-liquid extraction method to effectively extract pesticide components from the honey matrix and optimized analytical conditions to achieve high sensitivity and selectivity. The limits of detection (LOD) and the limits of quantitation (LOQ) were set in the range of 6-15 ng/mL and 19-44 ng/mL, respectively and the correlation coefficient (R²) was greater than 0.99, confirming good linearity. In addition, the intra-day recoveries for each pesticide were 75-104%, and the coefficient of variation (CV) was less than 20%, which met the guideline recommended by the Ministry of Food and Drug Safety. The LC-MS/MS method developed in this study is expected to be used as a multi-residue analysis method for 6 neonicotinoid pesticides in honey.
Nitrogen-doped carbon nanomaterials (N-CNMs) were prepared using Ni(NO3)2 as a catalyst in the laminar diffusion flame. Doping the structure of carbon nanomaterials (CNMs) with nitrogen can significantly change the characteristics of CNMs. The purpose of this research is to study the effect of adding ammonia ( NH3) on the evolution of CNMs structure in the laminar flame of ethylene. Raman analysis shows that the intensity ratio ( ID/IG) of the D-band and G-band of N-CNMs increases and then decreases after the addition of NH3. The intensity ratio is a maximum of 0.99, which has a good degree of disorder and defect density. The binding distribution of nitrogen was analyzed by X-ray photoelectron spectroscopy (XPS), and a correlation was found between the amount of nitrogen and the morphology of N-CNMs. Nitrogen atoms predominantly present in the forms of pyrrolic-N, pyridinic-N, graphitized-N and oxidized-N, with a doping ratio of nitrogen atoms reaching up to 2.44 at.%. This study found that smaller nickel (Ni) nanoparticles were the main catalysts for carbon nanotubes (CNTs), and their synthesis followed the ‘hollow growth mechanism’ and carbon nanofibers (CNFs) were synthesized from larger Ni nanoparticles according to the ‘solid growth mechanism’. Furthermore, a growth mechanism for the synthesis of bamboolike CNTs using a specific particle size of the Ni catalyst is proposed. It is noteworthy that the synthesis and modulation of high-performance N-CNMs by flame method represents a simple and efficient approach.
Pellino, a highly conserved E3 ubiquitin ligase, is known to mediate ubiquitination of phosphorylated Interleukin-1 receptor-related kinase (IRAK) homologs in Toll signaling pathway. To understand the immunological function of TmPellino, we screened the knockdown efficiency of TmPellino by injecting TmPellino-specific dsRNA into T. molitor larvae. Subsequently, we investigated the larval mortality and the tissue-specific expression patterns of antimicrobial peptide (AMP) genes against microbial challenges. Interestingly, the results indicate that the expression of many AMP genes was upregulated in the Malpighian tubules of TmPellino-silenced T. molitor larvae. This study may provide basic information to understand how Tmpellino regulates AMPs production in T. molitor.
Recently, it is demonstrate that the invertebrates have a immune memory, called Immune priming (IP). It was partially studied that the IP is mainly regulated by epigenetic modification. Here, to understand the IP on antimicrobial peptides (AMPs) production, we investigated larval mortality and time-dependent expression patterns of AMP genes in T. molitor larvae challenged with E. coli (two-times injection with a one-month interval). Interestingly, the results indicate that the higher and faster expression levels of most AMP genes were detected compared to the non-primed T. molitor larvae. Our results may used to improve the understanding of mechanisms of invertebrate immune memory.
Tumor necrosis factor receptor-associated factor (TRAF) is known to regulate antimicrobial peptides (AMPs) production in mammals. Here, to understand the immunological function of TmTRAF against microbial challenge, the induction patterns of TmTRAF against microbial infection was investigated by qRT-PCR in the whole-body and tissue of young larvae. In addition, the effects of TmTRAF RNAi on larval mortality and expression of 15 AMP genes in response to microbial infection were investigated. Our studies may help to understand the basic role of AMP production.
Tube, an intracellular protein of the Toll-pathway, forms a complex with Pelle and MyD88, and regulates a signal transduction to activate NF-κB in Drosophila. To understand the antimicrobial function of TmTube, the induction patterns of TmTube were investigated at 3, 6, 9, 12, and 24 h-post injection of pathogens into 10th to 12th instar larvae. In addition, we investigated the effects of TmTube RNAi on larval mortality and tissue specific AMP expression in response to microbial challenge. Our results will provide a basic information to elucidate the immunological function of TmTube
Pelle, a serine/threonine kinase, is an intracellular component of the Toll pathway and is involved in antimicrobial peptides (AMPs) production due to pathogenic infection. It is known that the Pelle phosphorylates Cactus and activates the NF-κB signaling pathway in Drosophila, but it is not studied in Tenebrio molitor. In this study we investigated the tissue-specific expression patterns of the Pelle following pathogenic infection at 3, 6, 9, 12, and 24 hours. Additionally, larval mortality and AMP expression against microbial injection were investigated in dsPelle-treated T. molitor larvae. Our results may help to understand the antimicrobial function of TmPelle.
In insects, the glutathione S-transferase is initiated in both the detoxification process and the protection of cellular membranes against oxidative damage. In this study, we identified the open reading frame (ORF) sequence of GST-iso1 and 2 from Tenebrio molitor (TmGST-iso1 and 2). To investigate the expression patterrns of TmGST-iso1 and 2 in response to herbicide, 0.06, 0.6, and 6 ㎍/㎕ of butachlor (FarmHannong, Seoul, South Korea) was challenged into T. molitor larvae, resulting that the TmGST-iso1 were highly induced at 3 and 24 h-post injection. Whereas, the highest expression of TmGST-iso2 was detected at 24 h after treatment. This study may contribute to basic information about the detoxifying activities of T. molitor.
It is well known that the JNK pathway regulates AMP production against pathogenic infection in both vertebrates and invertebrates. Tenebrio molitor hep (Tmhep) is an homolog of MAP kinase kinase in mammals. Here, we investigate the immunological function of Tmhep in responses in microbial infection using RNA interference technology. The results showed that silencing of Tmhep increased the larval mortality against microbial challenge, as well as reduced AMP production compared to the control group (dsEGFP-treated group). Conclusively, Tmhep plays an critical role in antimicrobial defense in T. molitor larvae.
This investigation was conducted three times (April, August, and October) from 2023 at five surveyed sites in Gonyangcheon estuarine wetland. A total of benthic macroinvertebrates belonging to 59 taxa 44 families 21 orders in 8 classes of 4 phyla were collected. The numbers of benthic macroinvertebrates species in each surveyed site were from 10 (St.1) to 41 (St.2). As a community indices analysis, the average DI is 0.78 (±0.12), H' is 1.6 (±0.68), R′ is 0.96 (±0.39), and J´ is 0.61 (±0.24). Community indices tended to decrease as the diversity and abundance decreased downstream. Total ecological score of benthic macroinvertebrates community (TESB), an average score of 35.47 (±15.86). The Habitat of Clithon retropictum and Ellobium chinense belonging to endangered species II, were identified at st.3~st.5.
In the Gucheon stream of Deogyusan National Park, a survey was conducted at three sites (six sites in total) in 2018 and 2021. A total of benthic macroinvertebrates of 107 taxa 46 families 13 orders in 6 classes of 5 phyla were found. According to the analysis of the appearance by year, a total of 87 species were surveyed in 2018, and a total of 89 species were surveyed in 2021. As a results to the functional feeding group analysis, Gathering-Collector (GC) accounted for a high species share (2018: 29.89%, 2021: 25.84%). In the functional habitat group analysis, Clinger (CL) accounted for a high species share (2018: 56.32%, 2021: 57.30%). Average annual community indices analysis, DI 0.38(±0.05), H′ 3.48(±0.19), R′ 3.23(±0.41), and J′ 0.85(±0.04) in 2018, and DI 0.55(±0.09), H′ 2.98(±0.31), R′ 2.91(±0.65), and J′ 0.70(±0.05) in 2021. As a results of the benthic macroinvertebrates index (BMI) analysis, indicating that all sites were classified as "very good," with average BMI values of 95.24(±2.84) in 2018 and 96.99(±1.13) in 2021.