The time-zone of pollinating activity relative to numbers released of Apis mellifera in the strawberry(Janghui var.) houses was together from 9A.M. to 4P. M., and the peak time of pollinating activity was 1P.M.. The effects on pollinating activity relative to the comb numbers in the honeybee hive released in the strawberry houses were ordered 5bee combs(11,000heads), 3bee combs(6,600heads) and 4bee combs(8,800heads). The rate of workers lost in A. mellifera hives with 5bee combs during the strawberry cultivating period were lower than those of 3bee combs and 4bee combs. The rates of fruit set by pollinating activity relative to the comb numbers in the honeybee hive released in the strawberry houses were same level with over 98%. The fruit qualities; number of seeds, sugar content and rate of normal fruit set were same level, but fruit weights was recorded 5bee combs with 40.8g, and 4bee combs and 3bee combs were showed with 37.8g. The marketing income of 5bee combs was 8% higher than that of 4bee combs and of 3bee combs, respectively.

The time-zone of pollinating activity relative to numbers released of Apis mellifera in the strawberry(Seolhyang var.) houses was together from 9A.M. to 4P. M., and the peak time of pollinating activity was 11A.M.. The effects on pollinating activity relative to the comb number in the honeybee hive released at the strawberry houses were ordered 5bee combs(11,000heads), 4bee combs (8,800heads) and 3bee combs (6,600heads). The rate of workers lost in A. mellifera hives with 5bee combs and 4bee combs during the strawberry cultivating period were lower than that of 3bee combs. The rates of fruit set by pollinating activity relative to the comb number in the honeybee hive released at the strawberry houses were same level with over 98%. The fruit qualities; No. of seeds, sugar content and rate of normal fruit set were same level, but fruit weights were ordered 5bee combs in 37.2g, 4bee combs in 35.6g and 3bee combs in 32.6g. The marketing incomes of 4bee combs and 5bee combs were 9% to 13% higher than that of 3bee combs, respectively.

The black soldier fly, Hermetia illucens (BSF), has a worldwide distribution in the tropics and warm temperate regions and is active in the Korea from May through October. This species colonize a wide variety of decomposing vegetable and animal matter and oviposit in a variety of decomposing materials. In this study, how the BSF molting, adult emergence and mating rate changed by seasonal condition at the artificial rearing system was investigated. The black soldier fly larvae and pupae were reared under laboratory condition (27℃, 60% R.H.). Under the laboratory condition, molting and adult emergence were not influenced by seasonal factors such as climate, radiation intensity. But it is known that the sunlight is the most important factor of the mating. In the previous study the time of day, temperature, and humidity is significantly correlated with oviposition and mating. The rearing of BSF throughout the year is restricted by sunlight. In this study, the data shows definitely different mating numbers throughout whole year. The time of the day and sunlight density are changed with season and it influence on artificial rearing. To culture the black soldier fly throughout the year in Korea needs a more deep study under the artificial rearing system.

A multiplex polymerase chain reaction (PCR) was developed for the simultaneous detection and differentiation among Nosema apis and Nosema ceranae in honeybee. Three sets of primers were selected from different genomic sequences to specifically amplify a 831 bp amplicon within the SSU rRNA gene, specific for both N. apis and N. ceranae (MSSR primer); a 375 bp amplicon within the SSU rRNA gene, specific for N. apis (NA primer); and a 1,131 bp amplicon within SSU rRNA gene, specific for N. ceranae (NC primer). Using the primers in conjunction (multiplex PCR) we were able to N. apis and N. ceranae and to differentiate between them. The sensitivity of this PCR assay was approximately 102 spores per milliliter. We proposed that the multiplex PCR was sensitive, specific and rapid tool that can serve as a useful differential diagnostic tool for detecting N. apis and N. ceranae in honeybee.

This study was conducted to investigate the distribution pattern, ecological characteristics and life cycle of the Black Soldier Fly (Hermetia illucens, BSF). The BSF was widely distributed throughout Korea. The insect was mainly found in the vicinity of and in cattle sheds, manure sheds, living waste dump grounds, and food waste dump grounds. Developmental characteristics of the BSF are as follows: the egg was long oval shaped of 887㎛ in the major axis and 190㎛ in the minor axis; it weighed 24㎍. Female oviposited ca. 1,000 eggs on average; eggs hatched in 81 hours under laboratory condition (27℃, 60% R.H.). The duration of the larval stage was approximately 15-20 days. The size of the last instar larvae was 21mm. The cuticle of the pupae gradually acquired red-brown color and the size of them was 19 mm. The pupal stage was shorter for females (16 days) than males (15 days). Adults were sized about 13-20mm long and black-colored. The life span of adult insects was 5-8 days for the first generation (June-July), 7-10 days for the second generation (Aug.-Sept.), and 13-18 days for the third generation (Sept.-Oct.). Mating started on the next day of emergence and actively occurred at the third day after emergence. Mating mostly occurred between 10:00 and 16:00 during which light intensity is highest. Egg-laying started on the third day and was most frequent from the fourth to the sixth day after emergence. Similar to mating time, females oviposited mostly between 10:00 and 16:00.

A novel beetle antimicrobial protein from stimulated Copris tripartitus and the corresponding gene were isolated in parallel through differential display-PCR and expression in Escherichia coli. To find cDNA clones responsible for bacteria resistance, the suppression subtractive hybridization and GeneFishing differentially expressed genes system were employed in the dung beetle, Copris tripartitus immunized with lipopolysaccaride. One cDNA clone from eight subtracted clones was selected through dot blot analysis and confirmed by northern blot analysis. The 516-bp, selected cDNA clone was determined by 5' and 3'rapid amplication of cDNA ends and cloned into the GST fusion expression vector pGEX-4T-1 for expression of the protein. The expressed protein was predicted 14.7 kDa and inhibited the growth of gram-negative bacteria such as Escherichia coli and Pseudomonas aeruginosa. These results implied that the expressed protein is related to immune defense mechanism against microorganism.

Protease from various sources have been studied biotecnologically. For biotechnological applications, one highly preferred enzyme is protease. There have been no reports of cloned genes encoding digestive proteases in the Laccotrephes japonenis, Ranatra unicolor, Muljarus japonicus. These insects are considered to be a predator of aquatic insects. RT-PCR was used to amplify cDNA fragments for digestive proteases from total RNA the hole body of the insects. The flanking sequences of the 5'- and 3'- end of the these genes were characterized by RACE-PCR. Sequence analysis showed that these genes contained complete ORF. The deduced amino acid sequences of these protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin-like serine proteinase, 54% to Triatonatoma infestans salivary trypsin. To generate Laccotrephes japonensis serine protease, the DNA fragment coding for serine protease is cloning into suttle vector pBACⅠ, named pBAC1-JG and infected to Spodoptera frugiperda (sf9) insect cell. The cDNA encoding JG was expressed as a 32-kDa polypeptide in baculovirus infected insect cells and the recombinant protein showed activity in the protease enzyme assay using gelatin as a substrate.

Attacin is an antibacterial protein that is secreted by fat body cells of insect larva in response to bacterial infection. A 949 bp cDNA encoding the antibacterial protein attacin was isolated by employing annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR) from immunized Papilio xuthus larvae. The attacin cDNAs encoded 250 amino acid residues open reading frame with 60 residues prepropeptide. The deduced amino acid sequence of P. xuthus attacin showed significant identities with other Lepidopteran attacins. The attacin transcript was detected at significant level after injection with bacterial lipopolysaccharide (LPS). The predicted mature attacin was expressed as soluble fusion protein efficiently in bacterial expression system. To increase productivity and solubility, attacin was translationally fused with thioredoxin (Trx) protein and expressed in E. coli cells that are highly sensitive to the mature attacin. The recombinant attacin exhibited antibacterial activity against Gram-negative bacteria.

The morecular cloning, gene structure, expression and enzyme activity of a serine-like proteas frome Laccotrephes Japonensis were examined. In this study, RT-PCR was used to amplify cDNA fragments for serine-like proteases from total RNA the hole body of Laccotrephes japonensis. The flanking sequences of the 5'- and 3'- end of the this gene were characterized by RACE-PCR. Sequence analysis showed that this gene contained an 963bp ORF encoding 321 amino acids. The deduced amino acid sequence of this protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin precuror LlsgP4, 54% to Triatonatoma infestans salivary trypsin. To generate Laccotrephes japonensis serine-like protease, the DNA fragment coding for serine protease is cloning into suttle vector pBACⅠ, named pBAC1-JG and infected to Spodoptera frugiperda (sf9) insect cell. The cDNA encoding JG was expressed as a 32-kDa polypeptide in baculovirus infected insect cells and the recombinant JG showed activity in the protease enzyme assay using gelatin as a substrate.

To find some antibacterial peptides responsible for bacterial resistance, we performed differential hybridization with total cDNA probes which synthesized from normal and immunized larvae. Thirteen individual cDNA transcripts were expressed differentially in a total 1,862 random cDNA clones. One of upregulated genes is a novel member of the insect defensin-like peptide(Coprisin), a family of antibacterial peptide. Northern blot analysis showed that Coprisin was up-regulated at 4h and reached the highest point level at 16h after injection of E.coli. The deduced amino acid sequence of Coprisin was composed of 80 amino acids with predicted molecular weight of 8.6 kDa and PI of 8.72. Comparison of the deduced amino acid mature portion of Coprisin with defensin-like peptide of other insect indicated that it has 79.1% and 67.4% identity with Anomala cuprea and Allomyrina dichotoma, respectively. To find antibacterial active region of Coprisin, we synthesized four peptides corresponding to amino acid residues 1V-43N-NH2(CopN1), 5-16(CopN2), 19-30(CopN3) and 31-43(CopN4) of coprisin having amidated amino acid residues at their Cterminal. A 12-mer amidated at its C-terminus, ACALHCIALRKK-NH2 (Ala19-Lys30-NH2) was synthesized based on the deduced amino acid sequence, assumed to be an active site sequence. This peptides showed antibacterial activity against E.coli, Staphylococcus aureus, MRSA, Psedomonas syringae, and Pectobacterium carotovorium. Modified 9-mer peptide, LRCIALRKK-NH2, showed strong antibacterial activity than mellitin peptide used as a positive control against gram-negative and gram-positive bacteria. This peptide showed no haemolytic activity and quite stable at 100℃ for several hours of incubation and in a wide pH range(pH2-12). Therefore, this peptide may be a good candidate for the development of new drug with potent antibacterial activity without cytotoxicity.

We have previously shown that the larvae of swallowtail butterfly, Papilio xuthus, exhibit substantial antibacterial activity in the hemolymph, upon challenging with bacterial lipopolysaccharide (LPS). Here we report the isolation and molecular characterization of several immune inducible genes that are specifically expressed by employing annealing control primer (ACP)-based GeneFishing polymerase chain reaction (PCR) from P. xuthus the larva. Using 120 arbitrary ACPs, we identified 24 differentially expressed genes (DEGs) that are up-regulated in response to injected LPS. Sequence analysis showed that 18 DEGs revelaed a high sequence similarity to the previously characterized genes of other insects, although 6 DEGs showed no significant similarity to any known genes. Among these inducible transcripts we found 8 putative immune-related genes including cecropin and attacin. Finally, we analysed the expression profiles of potential immune-related genes by RT-PCR and found all of them were considerably increased in the mRNA levels by LPS injection.

Protease from various sources have been studied biotecnologically. For biotechnological applications, one highly preferred enzyme is protease. There have been no reports of cloned genes encoding digestive proteases in the Laccotrephes japonenis, Ranatra unicolor, Muljarus japonicus. These insects are considered to be a predator of aquatic insects. RT-PCR was used to amplify cDNA fragments for digestive proteases from total RNA the hole body of the insects. The flanking sequences of the 5'- and 3'- end of the these genes were characterized by RACE-PCR. Sequence analysis showed that these genes contained complete ORF. The deduced amino acid sequences of these protease showed 62% identity to the serine protease of Creontiades dilutus, 58% to Lygus loneolaris trypsin-like serine proteinase , 54% to Triatonatoma infestans salivary trypsin. In the further study, to generate digestive protease, the DNA fragment coding for serine protease, trypsin-like serine protease were cloning into suttle vector pBACⅠ, and infected to Spodoptera frugiperda (sf9) insect cell. After that, we expect to carry out the proteolytic activity of these recombinant proteases. This is intended as a basis for future studies on the digestive protease in the insects.

The oxygen and nitrogen enriched activated carbons were obtained from modification of commercial activated carbon by using nitric acid, sodium hydroxide and urea. Zeta-potentials of modified activated carbons were investigated in relation to copper ion adsorption. The structural properties of modified activated carbons were not so much changed, but the zeta-potentials and isoelectric points were considerably changed. The zeta-potential of nitric acid modified activated carbon was the most negative than other activated carbons in the entire pH region, and the pHIEP was shifted from pH 4.8 to 2.6, resulted in the largest copper ion adsorption capacities compare with other activated carbons in the range of pH 3~6.5. In case of urea modified activated carbon, copper ion adsorption was larger than that of the as-received activated carbon from pH 2 to pH 6.5 even though the pHIEP was shifted to pH 6.0, it was due to the coordination process operated between nitrogen functional groups and copper ion. The adsorption capacity of copper ion was much influenced by zeta-potential and pHIEP of carbon adsorbent.

The present study aimed to examine the effect of dietary Ptecticu tenebrifer powder mixtures as pet dogfeed ingredients on crude fat and ash digestibility. Three groups of feeds Feed A, Feed B, and Feed C supplied from three farms were fed to a total of 45 dogs. The dietary Ptecticu tenebrifer powder mixture were prepared by mixing 25 g of Ptecticu tenebrifer powder with 100 g of canned food. Feed A, Feed B, and Feed C containing dietary Ptecticu tenebrifer powder mixtures were fed to 15 dogs of each breed of bichon, poodle, and chihuahua that were divided into three groups following a completely randomized design. For measuring the crude fat and crude ash digestibility, manure of each dog breed from each group were collected. Crude fat digestibility was not statistically significant among the dog breeds fed with feed C (p>0.05), but overall there was a statistical difference between the feed and the group by dog breed (p<0.05). In terms of crude ash digestibility, the three types of feed showed differences with respect to dog breeds (p<0.05). However, the group with no significant difference was observed in Feed B by dog breed (p>0.05). In conclusion, feeding Ptecticu tenebrifer powder mixture to dog breeds had no positive effect on the crude fat and ash digestibility and can be used as pet dogfeed ingredients.

This study aimed to investigate the effects of dietary Ptecticus tenebrifer on the fecal microbiomes of bichon frise. A total of 16 bichon frise dogs (average weight, 2 kg) were randomly allotted to 4 dietary treatments (4 dogs/group): general pet food, two types of domestic pet food containing Ptecticus tenebrifer, and one imported pet food containing Ptecticus tenebrifer. In the controls, Firmicutes accounted for the highest proportion (82%) at the phylum level in the fecal microbiomes. The Tax4Fun2's functional prediction program indicated that the control groups showed a relatively high amount of obesity-related microorganisms; the pathways included three types of carbohydrate metabolism. Among the treatments, Firmicutes abundances was the least in the treatments with the two types of domestic pet food containing Ptecticus tenebrifer; this did not affect the functional prediction of Tax4Fun2. In conclusion, the two types of domestic feed with Ptecticus tenebrifer were healthy and suitable for bichon frise; they could be beneficial in terms of obesity.

This study was conducted to investigate the effects of dietary supplementation with Ptecticus tenebrifer powder on the mortality and meat quality of Korean native chickens. A total of 40,000 Korean native chickens (1 day old, Hanhyup No. 3) were allocated to two dietary treatments (20,000 chickens in each treatment), which were fed the following: basal diet (control) and 1% of Ptecticus tenebrifer powder (T1). Feeding trials were conducted for 12 weeks, and mortality was measured weekly. At the end of the experimental period, 16 chickens (8 chickens in each treatment) were selected and slaughtered to obtain breast meat. The items used to analyze meat quality were pH, TBARS, and meat color. The weekly mortality rate was decreased by around 2 to 3 times in the T1 treatment group compared with the control group. The pH, TBARS, L*, and b* values of Korean native chicken breast were not affected by Ptecticus tenebrifer powder supplementation (p>0.05); however, a* values showed statistical significance (p<0.05). In conclusion, the addition of 1% Ptecticus tenebrifer powder reduced mortality rate and demonstrated its potential in livestock environmental management.

This study investigated the effects of dietary Ptecticu tenebrifer powder and canned mixtures on protein digestibility by different breeds of companion dogs (15 Bichons, 15 Malteses, 15 Chihuahuas and 15 Poodles). The mixtures were divided into Diet A, Diet B, Diet C, Diet D, and Diet E, which were supplied from five farms. Twenty-five grams each was mixed with 100 g of each canned food, and a total of 125 g was measured for each breed of dog. The result of component analysis of the mixtures showed the highest protein contents rather than dry matter, crude ash or crude fat. There were statistical significances (p<0.05) in all mixed feeds fed to bichon, maltese, chihuahua and poodle dog. Overall, protein digestibility by the breeds of dog ranged from 87.44% to 97.18%. Result of breed of dog comparison revealed that Diet E by poodle dog had the highest protein digestibility, and the lowest protein digestibility was observed in Diet C by Maltese. In conclusion, the use of dietary Ptecticu tenebrifer powder and canned mixtures did not only increased protein digestibility by different breeds of dog but also maintained normal manure properties.

We investigate the effects of the immune function (HI titer) in broilers fed diets containing Hermetia illucens (H. illucens) peptide extract over a 40-day period. Twenty-four broiler chicks (Arbor Acres, 1 d old) were divided into four groups and fed different diets (control, 0.1%, 0.5%, and 1% H. illucens peptide extract). To evaluate HI titer, all broilers were vaccinated with H9H2 vaccine subcutaneously on the lateral thorax, according to the manufacturer’s recommendations. Similar HI titer was observed with 1% H. illucens peptide extract treatment compared to the control after 40 days (p>0.01). Groups fed 0.5% H. illucens peptide extract demonstrated the most effective immune effects (p<0.01), followed by groups fed 0.1% H. illucens peptide extract. In conclusion, using 0.1% or 0.5% H. illucens peptide extract before or after vaccination improved HI titer immune function in broilers.

This study was conducted to evaluate the growth performance of broilers fed different levels of Hermetia illucens powder. A total of 400 broiler chicks (1-day old Arbor Acres) were fed commercial diets containing H. illucens powder at 0%, 0.1%, 0.5%, and 1% with four replicates (25 chicks per replicate), for 35 days. Weight gain in broilers fed diets containing different levels of H. illucens powder increased significantly at 28 and 35 days, compared with that of the control (p<0.05). However, feed intake and mortality showed no differences among the treatments as a function of time. At 21, 28, and 35 days, broilers fed different levels of H. illucens powder had lower feed conversion rates (p<0.05) than their counterparts fed the control diet. In conclusion, 0.5% H. illucens powder is the optimal level for improved weight gain and feed conversion.