Long-term non-surgical contraceptive methods for cats, especially community cats, are of global interest for cost-effective and humane reasons. This study aimed to investigate the effectiveness of a gonadotropin-releasing hormone (GnRH)-based vaccine for immunocontraception and to confirm its safety in intact female cats. Recombinant Salmonella typhimurium flagellin fljB (STF2)-GnRH protein was expressed in Escherichia coli. We divided female cats into vehicle control group (n = 4) and two experimental groups (100 μg injection group [n = 7] and 1000 μg injection group [n=7]), and immunized them twice intramuscularly (0.2 mL/cat at zero week and 4 weeks later into the other leg). Breeding trials started on day 120. All control cats (n = 4/4), 71% of the 100 μg injection group (n = 5/7), and 57% of the 1000 μg injection group (n = 4/7) became pregnant within 203 days after the introduction of male cats. The 1000 μg injection group had significantly a longer median time to conception following treatment (166 days) than the control (17 days, p < 0.05). Average litter size was significantly lower in the 1000 μg GnRH-vaccinated cats (2.8 ± 0.7) than in the control cats (4.5 ± 0.5, p < 0.05). Injection site reactions were not observed in any cat. The E. coli-expressed STF2-GnRH vaccine did not provide contraception in a sufficient proportion of the cats. However, it might be effective to suppress fertility through infertility vaccines before inducing permanent infertility through the trap-neuter-return.

World-class zoos have been recognized proponents of both animal welfare and wildlife conservation. The Korean zoos are in controversy over its existence, citing poor role performance. As one of the efforts to protect animals’ welfare and enhance wildlife conservation, leading zoos have been accredited through the inspections and evaluations. To improve the quality of Korean zoos, an accreditation standard that fits for Korean environment needs to be set. Out of various zoo accreditation standards around the world, we compared each category of all five publicly available accreditation standards; the Association of Zoos and Aquariums in the United States(AZA), Canada's Accredited Zoos and Aquarium(CAZA), European Association of Zoos and Aquaria(EAZA), Zoological Association of America(ZAA), and the Pan-African Association of Zoos and Aquariums(PAAZA). Based on the analysis result, we established 10 essential categories that were finally approved by the Focus group interview; animal welfare and care and management, veterinary care, conservation, education and interpretation, governing authority, staff, finance, physical facilities, safety and security, and master & strategy plan. This minimum zoo accreditation standards would enlighten the path to improve animal welfare and wildlife conservation of Korean local zoos.

Feral cats are widely considered to be leading the potential impacts on public health. This study aimed to provide estimates of vital data for feral cats relating Trap-Neuter-Return (TNR) to establish strategies effectively to manage feral cats in Pyeongtaek. Thus, this study focused on estimating feral cat population in Pyeongtaek and conducted a comparative analysis of the data for feral cats in Seoul (2013). The number of feral cats was estimated from 23,069 to 26,655 in Pyeongtaek, 2019. In relation to human population, when comparing the number of feral cats of Pyeongtaek and Seoul, it ranged from 4.57% to 5.28%, and from 1.97% to 2.55% respectively. This showed that Pyeongtaek was higher than Seoul. Fewer kittens were found in high-density areas, which the TNR project is believed to be generally effective in controlling the number of feral cats. In conclusion, in urban and rural complexes such as Pyeongtaek City, the number of feral cats compared to the population was higher than that of Seoul City, and the TNR program is believed to be somewhat effective in controlling the number of feral cats. When implementing TNR, it is necessary periodically to investigate the population and reflect them in policymaking.

Toll and IMD pathways play an important role in producing antimicrobial peptides (AMPs) through NF-κB in insects. The functions of IκB kinase (IKK) complex regulating the NF-κB signaling cascade have not yet been investigated in Tenebrio model. Here, we identified TmIKK-β (or TmIrd5) which contains 2,112 bp encoding 703 amino acid residues. Domain analysis shows that TmIKK-β contains one Serine/Threonine protein kinases catalytic domain. Developmental expression patterns indicate that TmIKK- β gene was highly expressed in early pupal (P1) and adult (A5) stages. Tissue specific profiles show that TmIKK-β was highly expressed in the integuments in last instar larvae, and fat body and hemocytes in 5 day-old adults. TmIKK-β1 transcripts were strongly induced at 3 and 12 h-post injection of E. coli, and 3 h-post injection of S. aureus or C. albicans in hemocytes. In gut, TmIKK-β transcripts were slightly induced by E. coli (at 6, 9 and 24 h) and C. albicans (at 24 h), while it was not induced by S. aureus challenge. Moreover, it was highly induced at 6 h-post injection of E. coli and then it was gradually decreased in the fat body. To understand the immunological role of TmIKK-β, gene specific RNAi and mortality assay was performed. Depletion of TmIKK-β mRNA leads to increase microbial susceptibility of larvae against E. coli, S. aureus and C. albicans. In addition, induction patterns of fourteen AMP genes in response to microbial challenge was tissue specifically investigated in TmIKK-β–silenced T. molitor larvae. The results suggest that expression of ten AMP genes out of fourteen genes were drastically decreased by TmIKK-β RNAi in fat body, suggesting that TmIKK-β plays an important role in antimicrobial innate immune responses.

Autophagy is an important self-eating process to eliminate damaged or unused organelles. We identified nine autophagy-related genes (Atg) including AaAtg-1, -3, -4b, -4d, -5, -6, -8, -12 and -13 from the Asian tiger mosquito, Aedes albopictus. Developmental expression patterns indicate that mRNA levels of AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 were highly expressed in egg, whereas expression of AaAtg8 was high in 1stand3rdinstarlarvalstages. TissuespecificexpressionofthesegenesindicatesthatAaAtg1 was highly expressed in thorax and midgut in blood-fed adult female mosquitoes (BF), and head and thorax in sugar fed adult female mosquitoes (SF). Transcript level of AaAtg3 was high in thorax in BF, but head, thorax and Malpighian tubules in SF. AaAtg4b, -4d mRNA levels were significantly high in Malpighian tubules in BF, and head in SF, respectively. AaAtg-5 and -6 transcripts were highly expressed in head in BF, and expression of AaAtg-8 was high in Malpighian tubules in BF. Levels of AaAtg-12 and -13 mRNAs were significantly high in head and midgut in BF. Induction patterns of AaAtg genes against pathogens showed that AaAtg-1, -3, -4b, -8, -12 and -13 were strongly induced at 6 h-post injection of S. aureus, and mRNA levels of AaAtg-1, -3 and -13 were significantly induced by E. coli challenge after 3 h-post injection in SF abdominal carcass. In SF midgut, AaAtg-1, -3, -4b, -4d, -5, -6, -12 and -13 transcripts were drastically induced at 9 h-injection of E. coli and S. aureus, while expression of AaAtg-8 was highly induced by S. aureus and C. albicans at 9 h-post injection. Each AaAtg gene was slightly induced by E. coli, S. aureus or C. albicans at different time points in abdominal carcass in BF. Interestingly, AaAtg-8 was not induced by microbial challenge. While eight other Atg genes except AaAtg-8 were highly influenced by S. aureus at 6 and 9 h-post injection, E. coli at 3 h-post-treatment, and 3, 6, and 9 h-post inoculation. In the future, we will characterize the functional roles of autophagy during mosquito-microbes interaction.

It has been well known that IKK-β, -ε and –γ play a pivotal role in IMD pathway. In this study, TmIKK-ε was identified and their functions in countering pathogenic infections were investigated. We identified TmIKK-ε gene which including 2,196 bp nucleotides (encoding 731 amino acid residues). Domain analysis of TmIKK-ε indicates that there is one Serine/Threonine protein kinases catalytic domain. TmIKK-ε gene was highly expressed in 2 day-old pupal stage and the expression was gradually decreased until 1 day-old adults. Then the expression was slightly increased until 4 day-old adult stage. Tissue specific expression of TmIKK-ε mRNA was high in the gut, integuments and hemocytes in last instar larvae, and fat body, Malpighian tubules and testis in 5-daysold adult. In hemocytes, TmIKK-ε was drastically induced by E. coli injection after 3 h and by S. aureus at 3 and 12 h-post injection. In gut, expression level of TmIKK-ε was high at 6 h-post injection of microbial injection. Expression of TmIKK-ε in fat body was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-ε, gene specific RNAi and mortality assay were performed. TmIKK-ε RNAi caused increased larval mortality against E. coli, not S. aureus and C. albicans. Finally, to investigate the induction patterns of Tenebrio fourteen AMP genes in response TmIKK-ε RNAi, three microorganisms were treated into TmIKK-ε-silenced T. molitor larvae. Nine out of fourteen AMP genes were not induced by microbial challenge in TmIKK-β dsRNA-injected group. Taken together, our results indicate that TmIKK-ε may regulates nine antimicrobial peptide genes in response to microbial challenge in T. molitor fat body.

Host defense against pathogen invasion highly relies on immune defense machinery that is controlled by the nuclear factor-κB (NF-κB) of transcription factors. The Toll pathway are well known as an insect innate immune mechanism to protect host itself from invaded pathogens. Basically, in the edible insect, Tenebrio molitor, the Toll pathway is primarily activated by polymeric Lys-type peptidoglycans (PGNs), and components of fungal cell walls, β-1,3-glucan. Based on the current studies, the tremendous study has been focused on recognition and subsequent activation of spätzle in haemolymph, hence, there is a grave gap for intracellular event. Herein, in order to understand intracellular event of Toll signaling pathway, the Dorsal gene were identified. Moreover, domain analyses of TmDorsal2 gene indicate that there are two major domains such as Rel homology domain (RHD), ig-like, plexins, and transcription factors (IPT) domains. Based on the achieved results, TmDorsal2 mRNA was highly expressed in 1-day old pupa. Furthermore, TmDorsal2 was highly expressed in Malpighian tubules and fat body in last instar larvae (LL), and likewise mainly expressed in Malpighian tubules during adult 5-day old period, also the lowest expression of TmDorsal2 was observed in gonads. Moreover, TmDorsal2 mRNA levels after infection with E. coli appreciably went up at 6 and 9h time points. To investigate the effects of TmDorsal2 RNAi on larval susceptibility against various pathogens namely E. coli, S. aureus or C.albicans, dsRNA of TmDorsal2 has been synthesized the larvae dissected after 24h. As a result, TmAttacin1a, 1b and 2, TmDefencine1 and 2, TmTenecin1, 2, 3 and 4, TmCecropin2, TmColeoptericin1 and 2, Thaumatin-like protein 1 and 2 markedly reduced in the gut after injecting all mentioned microbes. In contrast, TmTenecin 2, Thaumatin-like protein 1 and 2 strikingly increased after microbe injection in the fat body. Interestingly, the most AMPs gene expression in whole body experimental case were upregulated. On the horizon, we will investigate effects of TmDorsal1 RNAi on larval susceptibility against various pathogens. Taken together, our studies may aid to understand insect innate immunity.

IKK-γ is an essential protein to form IKK complex which regulate NF-κB. We identified TmIKK-γ (or TmKenny) gene which has 1,521 bp of nucleotides encoding 506 amino acid residues. Domain analysis of TmIKK-γ shows that there are one NF-κB essential modulator (NEMO) domain and a leucine zipper domain. Expression of TmIKK-γ gene was gradually increased from egg to 2-day-old pupal stage, dramatically decreased until 7 day-old pupal stage, and then it was gradually increased. TmIKK-γ transcripts were highly expressed in fat body and hemocytes in late instar larvae and integuments, fat body and Malpighian tubules in 5 day-old adult. TmIKK-γ was drastically induced by E. coli after 3 h challenges and by S. aureus at 3 and 12 h-post injection in hemocytes. TmIKK-γ was not induced by C. albicans although it was significantly induced by E. coli (at 3, 6 and 24 h) and S. aureus (at 9 h) in gut. In fat body, expression of TmIKK-γ was drastically induced by E. coli at 3 and 24 h-post injection while it was not significantly induced by S. aureus and C. albicans. To understand the immunological role of TmIKK-γ, gene specific RNAi and mortality assay was performed. larval mortality against microbial challenge was dramatically increased by TmIKK-γ RNAi. Furthermore, we investigate the tissue specific induction patterns of fourteen AMP genes in response TmIKK-γ dsRNA-treatment. In fat body, ten AMP genes out of fourteen was not significantly induced by microbial challenge in TmIKK-γ dsRNA-treated group. Based on these results, TmIKK-γ might play an important role in antimicrobial innate immune responses in Tenebrio molitor.

In this study, a relatively effective process is used to sterilize Escherichia coli on the surface of micro-sized calcium citrate powder using nitrogen and argon as process gases in a low-temperature vacuum plasma treatment. The purpose of this study is to confirm and to introduce the effectiveness of homogeneous surface treatment for the sterilization of fine inorganic powder by the rotatable low-temperature RF plasma system designed by ourselves. The results of the test using 3M petrifilm showed that there were no remarkable spots in the case of the surface of plasma treated powder, whereas the untreated powder showed many blue spots, which indicating that the E. coli was alive. After 5 days, in the same samples, the blue spots were seen to be larger and darker than before, while the plasma-treated powder showed no changes. The results from FE-SEM analysis showed that the E. coli was damaged and/or destroyed by reactive species generated in the plasma space, resulting in the E. coli being sterilized. Furthermore, the sterilization effects according to the selected parameters (N2 and Ar; flow rate 30 and 50 sccm) adapted in this study were mutually similar, regardless of such different process parameters, and this indicates that homogeneous treatment of powder surfaces could be more effective than conventional methods. Therefore, the plasma apparatus used in this study may be a practical method to use in a powerful sterilization process in powder-type food.

Background: The ginsenosides Rb1 (G-Rb1) and Rg1 (G-Rg1) are used as marker compounds, and are the principal bioactive compounds assessed in the quality control of white ginseng. This study was conducted to analyze white ginseng samples of different and to obtain useful data for the quality control of white ginseng. Methods and Results: The variation in the content of G-Rb1 and G-Rg1 was evaluated among 35 samples of 4-, 5-, and 6-year-old white ginseng. The content of both G-Rb1 and G-Rg1 did not significantly differ among ages, and the relative ratio of the maximum to the minimum content of these within ginseng of the same ages was more than two. However, the ratio of G-Rb1 to G-Rg1 content in the 5- and 6-year-old ginseng was significantly higher than that in the 4-year-old one. According to the ‘Ginseng industrial act’, the standard (w/w, %) minimum G-Rg1 and G-Rb1 content is 0.10% and 0.20% or more, respectively. Among the 35 samples examined, the content of G-Rg1 was found to be 0.124 - 0.399% with none being less than the standard level, while that of G-Rb1, was 0.147 - 0.595%, with 4 samples (11.4%) failing to meet the standard levels. The content of G-Rg1 and G-Rb1 did not show a constant relationship with the size of ginseng. Conclusions: In our study, the content of both G-Rg1 and G-Rb1 varied widely, and there was no significant difference among cultivation ages. The results of the present study might provide useful information for the quality control of raw ginseng and processed white ginseng using marker compound.

Background : Cordyceps militaris has been an wonder drug to anti-aging efficacy and called the three main drugs with ginseng and deer antler from the past. Cordycepin, cordycepic acid (d-mannitol) and adenosine are known as functional ingredients in Cordyceps militaris. Among them, cordycepin, the representative component, has been reported as antimicrobial substance containing immune enhancement, anti-cancer and anti-inflammatory effects. Methods and Results : After Cordyceps militaris produced from different types of medium mixed with 10-fold volume of purified water, the mixture were extracted at 70±5℃ for a hour and that extracts re-extracted using ultrasonics wave for 30 minutes. Qualitative analysis of the index component was determined by using the Q-TOF (A quadrupole time-of-flight mass spectrometer), and quantitative analysis was performed by using HPLC (high-performance liquid chromatography) with Xselect HSS T3 column (2.1 X 100 mm, 2.5㎛, Waters, USA) and ultrapure water and acetonitrile as mobile phase A and B. Detection column temperature, injection volume and the flow rate were 35℃, 2 μL and 0.3 mL / minute respectively. The cordycepin content of Cordyceps militaris produced from medium mixed with vegetable and animal ingredients higher than single ingredient. Moreover, through a variety of analyzes by varying the type and content of the medium additives, the cordycepin in Cordyceps militaris produced from medium mixed with animal ingredients highest. Furthermore, the cordycepin content of a fruit body was higher than those of the a mycelium. Conclusion : These results provide a method for producing an high cordycepin content of Cordyceps militaris as functional food ingredient.

This study was carried out to investigate change of ginsenoside contents in red and fresh ginseng according toroot part and age by hydrolysis. Neutral total ginsenoside contents by hydrolysis in 6-year main root and lateralroot were significantly increased than those by non-hydrolysis, as 41.6 and 32.8%, respectively. However, there wasno significant difference in red ginseng. In fresh ginseng, ginsenoside contents of the protopanaxatriol group such as Re, Rf,Rg₁, Rg₂, and Rh₁ were not significantly different, but Rb₁, Rb₂, Rb₃, Rc, and Rd showed significant difference. The increaserate of neutral total ginsenoside content by hydrolysis was higher in epidermis-cortex than stele. Also, the neutraltotal ginsenoside content was fine root>rhizome>lateral root>main root, respectively. While there was no tendencytowards the increase of ginsenoside by hydrolysis with the increase of root age in fine root and rhizome, there was significantdecrease in main root and lateral root.

This study was carried out to investigate the correlation between root diameter and ginsenoside compositionof Panax ginseng C. A. Meyer cultivar Yunpoong. Dry matter ratio of main root was a little higher than that of lateral rootand fine root, and that was higher by the increase of root diameter in the same root parts. Total ginsenosides composition ofmain and lateral roots increased by the decrease of root diameter, especially in lateral root. Similar resulted in fine root, butthere was no significant difference where root diameter was below 2.5㎜. Except for ginsenoside-Rg1, other ginsenosidescomponent, PDs, PTs and total ginsenosides had highly negative correlation with the root diameter within whole root, mainroot+lateral root and lateral root+fine root, while Rg1 had positive correlation with the root diameter.