Various arthropods have directly and indirectly threatened human life by transmitting human pathogens such as malaria and Zika virus as well as by damaging crops such as direct feeding and transmission of plant pathogens. According to WHO, vector-borne diseases account for over 17% of all infectious diseases and there are over 1 million deaths from the diseases such as malaria, dengue, etc., globally. About 13% of total crop production is destroyed by agricultural pests during crop production and storage. To control medical or agricultural pests chemical pesticides have been used, but recently because of concerns about environmental pollution and human health the demands for eco-friendly control method are increasing. Insect pathogens are good alternative candidates instead of chemicals. Over 50 entomopathogens including viruses, bacteria, fungi and nematodes are now commercially used as microbial pesticides. Entomopathogenic fungi have also been evaluated as control agents for a diverse insect pests such as aphid, moth, thrip, whitefly, mosquitoes, tick, etc. We will discuss a development of microbial pesticides using, especially, entomopathogenic fungi to control noxious arthropods to agricultural crops and human.

Even though klotho deficiency in mice exhibits multiple aging-like phenotypes, studies using large animal models such as pigs, which have many similarities to humans, have been limited due to the absence of cell lines or animal models. The objective of this study was to generate homozygous klotho knockout porcine cell lines and cloned embryos. A CRISPR sgRNA specific for the klotho gene was designed and sgRNA (targeting exon 3 of klotho) and Cas9 RNPs were transfected into porcine fibroblasts. The transfected fibroblasts were then used for single cell colony formation and 9 single cell–derived colonies were established. In a T7 endonuclease I mutation assay, 5 colonies (#3, #4, #5, #7 and #9) were confirmed as mutated. These 5 colonies were subsequently analyzed by deep sequencing for determination of homozygous mutated colonies and 4 (#3, #4, #5 and #9) from 5 colonies contained homozygous modifications. Somatic cell nuclear transfer was performed to generate homozygous klotho knockout cloned embryos by using one homozygous mutation colony (#9); the cleavage and blastocyst formation rates were 72.0% and 8.3%, respectively. Two cloned embryos derived from a homozygous klotho knockout cell line (#9) were subjected to deep sequencing and they showed the same mutation pattern as the donor cell line. In conclusion, we produced homozygous klotho knockout porcine embryos cloned from genome-edited porcine fibroblasts.

Beet armyworm, Spodoptera exigua, diamondback moth, Plutella xylostella and tobacco cutworm, Spodoptera litura are the three most serious pests of many economically important crops such as cruciferous crops, various vegetables and ornamental plants. Because these pests are known to be resistant to lots of chemical insecticides, integrated control using both or either entomophathogen and/or natural enemy is thought to be an attractive alternative for effective control. One of the obstacles using and expanding mycopesticide is narrow host ranges. At commercial farms, cultivating crops are seriously damaged by various Lepidopteran pests. Farmers want to use a microbial control agent which can control various host insects to reduce cost and labor. In previous study, we selected two entomopathogenic fungi, Metarhizium anisopliae and Paecilomyces fumosoroseus, which shown high virulence against beet armyworm. For wide use of the isolates in farm, we tested its host ranges, especially to diamondback moth and tobacco cutworm, which also are serious pest in Korea. The two isolates were shown a good control effect in leaf disc bioassy.

Various insect pests and plant disease can outbreak in a field. For the effective control of pests and plant diseases during crop cultivation, farmers simultaneously or sequentially spray various eco-friendly agricultural materials (EFAM), chemical pesticides and microbial control agents on the same fields. It was reported that many agrochemicals are harmful to entomopathogenic fungi, especially some fungicides with broad spectrum activity that are routinely applied for the control of plant diseases. In addition, some pesticides may antagonize the potential insecticidal activity and efficiency of entomopathogenic fungi. Therefore, sometimes the utilization of fungal entomopathogen in forestry and agricultural production is limited because of the undesirable interference from some fungicides and pesticides. There is little research that examines the compatibility of these EFAMs with entomopathogenic fungi and the influence of EFAMs on the control efficacy of mycopesticides. We conducted a study of influence of pretreated eco-friendly agricultural materials on control efficacy of Isaria javanica isolate against sweet potato whitefly.

Microbe have been considered as potential control agents for pest, as alternative to chemical control methods. Among entomopathogens, fungi cause the mortality by penetrating the cuticle of pest and/or by metabolites such as toxin. Not only this direct control effect of fungi, but repellency of fungi also may be used to prevent the pest. Repellence effect of fungi is considered as inhibitory factor to control termite. A study was reported in Japan that termite was able to detect and remove the conidia of fungi on their surface. The termite can escape from fungal infection and protect their colony. There is few study that insect pest such as moth can detect and avoid the virulence fungi. Therefore, we has been conducting the detection and avoidance of beet armyworm to high pathogenic fungi, Paecilomyces fumosoroseus. Adult of the beet armyworm avoided oviposition at Chinese cabbage treated with P. fumosoroseus compare to control. This result may be used to prevent the infestation of moth in crop production.

Entomopathogenic fungus is a useful control agent to sucking type insect such as whitefly and aphid. The fungi are influenced by some environmental factors such as relative humidity, temperature and UV and cause slow and fluctuation in pest control efficacy. Especially, UV kills conidia or spores of entomopathogenic fungi and a mycopesticide using fungi has short control period in field. UV intensity changes from season to season. Survival rate of entomopathognic fungi treated may differ from seasons and will show different control efficacy. Therefore, we conducted a study to estimate the persistence of an Isaria javanica isolate, which was already reported as sweet potato whitefly control agent, in potted greenhouse soil planted different crops. The number of survival spore decreased gradually and differ from seasons.

As ubiquitous organisms entomopathogenic fungi may natually occur within environment including phylloplanes and soil or deposite and survive for a while after spraying a fungal suspension to control insect pest. The existence of pathogenic fungi is a threat to arthropods foraging for food and oviposition sites. The detection and avoidance of pathogens is important for host survival, longevity and ultimate fitness. Although entomopathogenic fungi are ubiquitous worldwide and act as common natural enemies of many invertebrate species, very few studies have conducted abiut the detection and avoidance of pathogenic fungi by insect pest. We have studied that an insect pest can recognize a high virulence fungi on crop surface, avoid oviposition and don’t enter the field treated their pathogen.

Entomopathogenic fungi are natural enemies of insect pests and contribute to the natural regulation of their host populations. These fungal group are often used as active ingredients for microbial insect pest control. In addition, the potential antimicrobial effect by entomopathogenic fungi including Beauveria bassiana, Lecanicillium spp., and Isaria fumosorosea have recently been reported against fungal plant pathogens. Dual microbial control effects with entomopathogenic fungi against both aphids and cucumber powdery mildew had reported in Canada. In our previous studies we conducted bioassay with entomopathogenic fungi to develop dual microbial control agent which can control both aphid and fungal plant disease. We selected an Beauveria bassiana isolate which has high dual control effects against both cotton aphid, Aphis gossypii and sclerotinia rot, Sclerotinia sclerotiorum. In this study, we have tested the dual control efficacy of the B. bassiana isolate against cotton aphid and sclerotinia rot on whole potted cucumber plants. We found that the B. bassiana isolate protected the plant from cotton aphid and sclerotinia rot under laboratory condition.

Insect pests have been a serious problem over many years and remain a major threat for food production. Although chemical pesticides are major pest control strategies, use of microorganisms such as entomopathogenic bacteria, fungi, viruses and nematodes have continuously increase last few decades to minimize the use of agrichemicals. According to BBC research, the global biocontrol market was about $2.1 billion in 2011, and this is expected to rise $3 ~ 4 billion by 2017. Over 50 entomopathogens are commercially produced and used augmentatively as microbial pesticides. About 175 biopesticide active ingredients and 700 products have been registered worldwide. Bacillus thuringiensis (Bt), Beauveria bassiana, Metarhizium spp., nuclear polyhedrosis virus and Steinernema spp. are the most popular control agents used in plant protection. Among the microbial control agents Bt products have more than 50% of market share. In Korea, only 13 environmentally-friendly crop protectants were registered for insect pest control in 2015. Market share is very low and has grown slowly. We will discuss how we can expand the market with our techniques.

Acetylcholinesterase (AChE) is a hydrolase that hydrolyzes the neurotransmitter acetylcholine. Soluble form of AChE is generated via alternative splicing and functions as a bioscavenger in Dropsophila melanogaster. In this study, effects of acetic acid on the soluble AChE expression were investigated. Treatment of acetic acid resulted in over-expression of soluble AChE in the abdomen in a dose-dependent manner. The soluble AChE was determined to be expressed in the fat body. However, no apparent change in AChE expression was observed in the head. Our finding suggests that the soluble AChE is involved in chemical defense against high concentration of acetic acid, which is a common by-product in fermenting foods. The high level of acetic acid resistance in D. melanogaster, thus, appears to have been evolved via the induction mechanism of soluble AChE expression.

This study is the first report about the toxicity of pesticides to the mycophagous predator, I. koebelei, of powdery mildew of agricultural crops. Pesticides we tested are composed of synthetic and environmental-friendly products and being used conventionally for the control of insect or microbial pests on cucumber in Korea. our study was conducted to determine the relative toxicities of several pesticides used in Korea cucumber production to mycophagous natural enemy, I. koebelei and to provide a background for implementation of integrated powdery mildew management programs. Based on IOBC classification, three insecticides, bifenthrin + imidacloprid, acetamiprid + indoxacarb, acetamiprid + etopheprox are classified as having a Class 4 (harmful). Spiromesifen showed the low toxicity to the survival and the fecundity of I. koebelei when this chemical had been exposed to 3rd larva or newly emerged adult via feeding with cucumber powdery mildew. However, pyriproxyfen not only decreased the fecundity of female adult but also strongly prohibited from pupation. Many commercial biological or botanical pesticides can restrict the population of I. koebelei. However, Q pact (a.i. Ampelomyces quisqualis 94013), Top seed (a.i. Paenibacillus polymyxa AC-1), BT one (Bacillus thuringiensis) and Solbitchae (insecticidal microorganism) had no toxicity to I. koebelei when this chemical had been exposed to 3rd larva or newly emerged adult feeding with cucumber powdery mildew.

Extracts from Aloe vera leaves, Aloe arborescens leaves, Aloe vera callus, Portulaca oleracea and cacao (Theobroma cacao L.) bean husk (CBH) were prepared using acetone, chloroform, ethanol, hexane, and water. Solvent extracts of Aloe vera leaf had very high antioxidant activities showing IC50 values in the ranges of 0.02-0.17 mg/ mL, and had the highest total phenolic and flavonoid content among the tested samples. We hypothesized that Aloe vera leaf and CBH extracts might possess considerable in vitro anti-glycation activities. Indeed, these extracts strongly inhibited the formation of advanced glycation end-products from RNase in the presence of ribose. The chloroform extract of Aloe vera leaf showed the strongest inhibition of AGE formation (99.9%), followed by the 95% acetone extract (92.8%) at a concentration of 1 mg/mL, exhibiting higher anti-glycation activities than those of AG and rutin (73.4% and 96.1% at 1 mg/mL, respectively). The anti-glycation activity of all extracts was correlated positively with their total contents of phenolics and flavonoids. We conclude that Aloe vera leaf extracts and their constituents may be used as anti-glycation agents.

Recently, methods that usea carbon-based filler, a conductive nanomaterial, have been investigated to develop composite fillers containing dielectric materials. In this study, we added geometric changes to a carbon fiber, a typical carbon-based filler material, by differentiating the orientation angle and the number of plies of the fiber. We also studied the electrical and electromagnetic shield characteristics. Based on the orientation angle of 0˚, the orientation angle of the carbon fiber was changed between 0, 15, 30, 45, and 90˚, and 2, 4, and 6 plies were stacked for each orientation angle. The maximum effect was found when the orientation angle was 90˚, which was perpendicular to the electromagnetic wave flow, as compared to 0˚, in which case the electrical resistance was small. Therefore, it is verified that the orientation angle has more of an effect on the electromagnetic interference shield performance than the number of plies.