Development of an ELISA for the detection of the residues of the fungicide iprovalicarb

A competitive indirect enzyme-linked immunosorbent assay was developed for the fungicide iprovalicarb, using a polyclonal antibody produced against a hapten conjugated through the carboxyl group on the benzene ring to keyhole limpet hemocyanin. Under an optimized condition using a heterologous format, an IC(50) of 3.51 ng/mL and the lowest detection limit of 0.065 ng/mL were obtained. When the isopropoxy group was removed from the iprovalicarb structure for the synthesis of a hapten, the resulting hapten was not successful as an immunogen, indicating that the isopropyl moiety was an important epitope, as evidenced by the cross-reactivities of some structurally related compounds. When applied to the real crop and water samples, the recoveries were in the range of 80.52-144.70% (n = 4) and 72.11-100.43% (n = 4), respectively. Accordingly, this ELISA can be used as a useful method for monitoring iprovalicarb residues in crop and water samples.     

Development of an enzyme-linked immunosorbent assay for the detection of the fungicide fenarimol

To develop an enzyme-linked immunosorbent assay for the fungicide fenarimol, two synthesized haptens, haptens-1 and -2, and the purchased 4,4'-DDA were conjugated to carrier proteins (BSA, KLH, and OVA). Polyclonal antibodies raised against hapten-1,2-KLH conjugates in rabbits and the coating antigens of hapten-1,2-BSA conjugates, hapten-2-OVA conjugate, and 4,4'-DDA-BSA conjugate were screened and selected for the homologous and/or heterologous ELISA formats. Two competitive indirect ELISAs were selected: assays I and II. The optimized ciELISAs of assays I and II showed average IC(50) values of fenarimol of 5.4 and 9.4 ng/mL, detection ranges of 1.1-25.9 and 1.1-82.7 ng/mL, and lowest detection limits of 0.3 and 0.3 ng/mL, respectively. The cross-reactivities with several structurally related compounds indicated the importance of the steric fitness in the antigen-antibody interaction. Recoveries of fenarimol from apple and pear samples spiked with the analyte by assay I were in the range of 93-113% by simple extraction, concentration, and dilution. This assay could be a convenient and supplemental analytical tool for monitoring fenarimol residues in environmental and agricultural samples.     

Preparation, characteristics, and stability of glutathione-loaded nanoparticles

The aim of this study was to investigate the characteristics and oxidative stability of chitosan-glutathione conjugate (CS-GSH) and CS-GSH nanoparticles (CS-GSH NPs) to explore the potentials of these nanoparticle systems for GSH delivery. CS-GSH was synthesized using a radical polymerization method, and CS-GSH NP was prepared by ionic gelation of CS-GSH with sodium tripolyphosphate (TPP). The sizes of CS-GSH NPs significantly increased with increasing CS-GSH concentration and CS-GSH/TPP ratio. The entrapment efficiency (EE) significantly increased with increasing CS-GSH concentration and significantly decreased with increasing CS-GSH/TPP ratio. The immobilized GSH could be protected against oxidation compared to free GSH. The thiol content in the nanoencapsulated GSH was more effectively maintained than those in free GSH and CS-GSH, regardless of the presence of oxidative stress-inducing agents. These results suggest that CS-GSH NP can be used to enhance the oxidative stability of GSH.     

Ribosomal protein s3 immunoreactivity in the young, adult and aged gerbil hippocampus

Ribosomal protein S3 (rpS3) is well known to participate in DNA repair mechanisms. In the present study, we compared rpS3 immunoreactivity and its protein levels in the hippocampus among young, adult and aged gerbils. In the postnatal month (PM) 3 group as the young, rpS3 immunoreaction was observed in pyramidal and non-pyramidal cells of the hippocampus proper and in granule and polymorphic cells of the dentate gyrus. In the PM 12 group as adult and 24 group as the aged, rpS immunoreactivity in the hippocampus was decreased compared to the PM 3 group. Western blot analysis showed that rpS3 levels were decreased with time; lowest at PM 24. These results indicate that rpS3 immunoreactivity and protein levels were markedly decreased in the aged gerbil hippocampus.     

Determination of staphylococcal exotoxins, SCCmec types, and genetic relatedness of Staphylococcus intermedius group isolates from veterinary staff, companion animals, and hospital environments in Korea

The Staphylococcus (S.) intermedius group (SIG) has been a main research subject in recent years. S. pseudintermedius causes pyoderma and otitis in companion animals as well as foodborne diseases. To prevent SIG-associated infection and disease outbreaks, identification of both staphylococcal exotoxins and staphylococcal cassette chromosome mec (SCCmec) types among SIG isolates may be helpful. In this study, it was found that a single isolate (one out of 178 SIG isolates examined) harbored the canine enterotoxin SEC gene. However, the S. intermedius exfoliative toxin gene was found in 166 SIG isolates although the S. aureus-derived exfoliative toxin genes, such as eta, etb and etd, were not detected. SCCmec typing resulted in classifying one isolate as SCCmec type IV, 41 isolates as type V (including three S. intermedius isolates), and 10 isolates as non-classifiable. Genetic relatedness of all S. pseudintermedius isolates recovered from veterinary staff, companion animals, and hospital environments was determined by pulsed-field gel electrophoresis. Strains having the same band patterns were detected in S. pseudintermedius isolates collected at 13 and 18 months, suggesting possible colonization and/or expansion of a specific S. pseudintermedius strain in a veterinary hospital.     

Field comparison of the interferon-gamma assay and the intradermal tuberculin test for the diagnosis of bovine tuberculosis

An extensive field comparison of the gamma interferon (IFN-gamma) assay and the single intradermal tuberculin test for the diagnosis of bovine tuberculosis was conducted in Australia. The specificity of the IFN-gamma assay was determined by testing more than 6000 cattle from tuberculosis-free herds and varied from 96.2% to 98.1%, depending on the cut-off point chosen to define a positive reactor. For the sensitivity trial, cattle from herds being de-populated because of bovine tuberculosis were examined with both assays. The sensitivity of the IFN-gamma assay was shown to be significantly higher than the single intradermal tuberculin test and varied from 76.8% to 93.6% depending on the method of interpretation. A maximum overall sensitivity of 95.2% was obtained by testing with the IFN-gamma and the tuberculin test in parallel. The superior sensitivity of the IFN-gamma assay and the ability to adjust the sensitivity of the system depending on the task involved, will provide the Australian Tuberculosis Eradication Campaign with a valuable additional test to enable it to accomplish its goals.     

Effects of Dietary Chlorella ellipsoidea Supplementation on Growth, Blood Characteristics, and Whole-Body Composition in Juvenile Japanese Flounder Paralichthys olivaceus

The study was conducted to investigate the effects of dietary Chlorella powder (C) supplementation on growth performance, blood characteristics, and whole-body composition in juvenile Japanese flounder Paralichthys olivaceus . Four experimental diets were supplemented with C at 0, 1, 2 and 4% (C₀, C₁, C₂, and C₄) on a dry-weight basis. Three replicate groups of fish averaging 1.13 ± 0.02 g (Mean ± SD) were randomly distributed in each aquarium and fed one of four experimental diets for 12 wk. After 12 wk of the feeding trial, fish fed C₂ diet had higher weight gain (WG), feed efficiency (FE), and protein efficiency ratio (PER) than did fish fed C₀ and C₁ diets ( P 0.05); however, there was no significant difference among fish fed C₁ and C₄ diets, and among fish fed C₂ and C₄ diets ( P > 0.05). Fish fed C₂ and C₄ diets had a lower serum cholesterol level than did fish fed C₀ and C₁ diets ( P 0.05). Fish fed C₂ and C₄ diets had a lower body fat than did fish fed C₀ ( P 0.05). These results indicate that dietary supplementation of 2% Chlorella powder in the commercial diets could improve growth, feed utilization, serum cholesterol level, and whole-body fat contents in juvenile Japanese flounder.     

Age-related changes in ionized calcium-binding adapter molecule 1 immunoreactivity and protein level in the gerbil hippocampal CA1 region

Microglia are evenly distributed throughout the brain parenchyma. They respond rapidly to a variety of alterations in the microenvironment of the brain and act as sensors for pathological events in the brain. In the present study, we investigated the age-dependent changes in the immunoreactivity and protein level of ionized calcium-binding adapter molecule 1 (Iba-1), a microglial marker, in the CA1 region of the gerbil hippocampus. Iba-1 immunoreactive microglia were detected in the hippocampal CA1 region of the postnatal month 1 (PM 1) group. Iba-1 positive microglia were morphologically inactive between the PM 1 and PM 12 stages. Some Iba-1 immunoreactive microglia were present in the active form in the hippocampal CA1 region of the PM 18 and PM 24 groups. The Iba-1 protein levels in hippocampal CA1 homogenates were decreased in the PM 1 through PM 6 groups and increased in an age-dependent manner thereafter. These results suggest that Iba-1 immunoreactive microglia in the active form were detected in the hippocampal CA1 region in the PM 18 and PM 24 groups. This result may be associated with an age-dependent susceptibility to neurodegenerative diseases associated with the hippocampus.     

Intrapancreatic ectopic splenic tissue found in a cloned miniature pig

Somatic cell nuclear transfer (SCNT) is a cost-effective technique for producing transgenic pigs. However, abnormalities in the cloned pigs might prevent use these animals for clinical applications or disease modeling. In the present study, we generated several cloned pigs. One of the pigs was found to have intrapancreatic ectopic splenic tissue during histopathology analysis although this animal was grossly normal and genetically identical to the other cloned pigs. Ectopic splenic tissue in the pancreas is very rare, especially in animals. To the best of our knowledge, this is the first such report for cloned pigs.