• Journal of Food Hygiene and Safety
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• v.5 no.3
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• pp.159-164
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• 1990

To detect and quantitation residual antibiotics and antibacterial agents in meats, we performed a biological assay employing the three microorganisms Bacillus subtilis ATCC 6633, Micrococcus luteus ATCC 9341, and Bacillus cereus var. mycoides ATCC 11778 for the screening purpose and developed a Gas Chromatography-mass Spectrometry(GC/MS) analysis for the confirmation and quantiation. In the biological assay (paper disk method), three test solution are used depending on the character of the residual antibiotics and antibacterial agents, follow by a simple clean up procedure which includes homogenization with Mcilvaine buffer, defatting with includes homogenization with Mcilvaine buffer, defatting with hexane, extraction with chloroform, clean-up by Sep-Pak $C_{18}$ and Bakerbond SPE carboxylic acid column. The chloroform layer is used for the analysis of sulfa agents. macrolides antibiotics and antibacterial agents, Adsorbed materials in the Sep-Pak $C_{18}$ were also employed for th analysis of penicillins and tetracyclines. Effluents from the Sep-Pak $C_{18}$ were cleaned-up one more by Bakerbond 10 SPE COOH column and employed for the analysis of aminoglycosides. In the instrumental analysis by using the GC/MSD, residual antibiotics and antibacterial agent were quantitated by selected ion monitoring (SIM) mode after derivatization. A simultaneous analysis of six residual antibiotic and antibacterial agent such as oxytetracycline, penicillin, ampicillin, choliraphenicol and thiamphenicol was developed with simple cleanup procedures revealing good recovery and reproducibility. Also, simultaneous detection of macrolides antibiotics such as erythromycin, spiramycin, and oleandomycin was developed after acid hydrolysis due to their large molecular structures. Because of the high reproducibility and selectivity of these two methods, it is very desirable that the combination of the two methods be used in the bioassay for the screening of residual antibiotics and antibacterial agent and that GC/MSD analysis be used for the confirmation and quantitation.

• Toxicological Research
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• v.34 no.1
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• pp.1-6
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• 2018

The purpose of this study was to detect cell death in the liver of mice treated with thioacetamide (TAA) using fluorescence bioimaging and compare this outcome with that using conventional histopathological examination. At 6 weeks of age, 24 mice were randomly divided into three groups: group 1 (G1), control group; group 2 (G2), fluorescence probe control group; group 3 (G3), TAA-treated group. G3 mice were treated with TAA. Twenty-two hours after TAA treatment, G2 and G3 mice were treated with Annexin-Vivo 750. Fluorescence in vivo bioimaging was performed by fluorescence molecular tomography at two hours after Annexin-Vivo 750 treatment, and fluorescence ex vivo bioimaging of the liver was performed. Liver damage was validated by histopathological examination. In vivo bioimaging showed that the fluorescence intensity was increased in the right upper part of G3 mice compared with that in G2 mice, whereas G1 mice showed no signal. Additionally ex vivo bioimaging showed that the fluorescence intensity was significantly increased in the livers of G3 mice compared with those in G1 or G2 mice (p < 0.05). Histopathological examination of the liver showed no cell death in G1 and G2 mice. However, in G3 mice, there was destruction of hepatocytes and increased cell death. Terminal deoxynucleotidyl transferase dUTP nick end labeling staining confirmed many cell death features in the liver of G3 mice, whereas no pathological findings were observed in the liver of G1 and G2 mice. Taken together, fluorescence bioimaging in this study showed the detection of cell death and made it possible to quantify the level of cell death in male mice. The outcome was correlated with conventional biomedical examination. As it was difficult to differentiate histological location by fluorescent bioimaging, it is necessary to develop specific fluorescent dyes for monitoring hepatic disease progression and to exploit new bioimaging techniques without dye-labeling.

• Parasites, Hosts and Diseases
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• v.48 no.1
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• pp.43-48
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• 2010

Cryptosporidium and Giardia are 2 protozoan parasites responsible for waterborne diseases outbreaks worldwide. In order to assess the prevalence of these protozoans in drinking water samples in the northern part of Portugal and the risk of human infection, we have established a long term program aiming at pinpointing the sources of surface water, drinking water, and environmental contamination, working with the water-supply industry. Total 43 sources of drinking water samples were selected, and a total of 167 samples were analyzed using the Method 1623. Sensitivity assays regarding the genetic characterization by PCR and sequencing of the genes, 18S SSU rRNA, for Cryptosporidium spp. and $\beta$, -giardin for G. duodenalis were set in the laboratory. According to the defined criteria, molecular analysis was performed over 4 samples. Environmental stages of the protozoa were detected in 25.7% (43 out of 167) of the water samples, 8.4% (14 out of 167) with cysts of Giardia, 10.2% (17 out of 167) with oocysts of Cryptosporidium and 7.2% (12 out of 167) for both species. The mean concentrations were 0.1-12.7 oocysts of Cryptosporidium spp. per 10 L and 0.1-108.3 cysts of Giardia duodenalis per 10 L. Our results suggest that the efficiency in drinking water plants must be ameliorated in their efficiency in reducing the levels of contamination. We suggest the implementation of systematic monitoring programs for both protozoa. To authors' knowledge, this is the first report evaluating the concentration of environmental stages of Cryptosporidium and Giardia in drinking water samples in the northern part of Portugal.

• Parasites, Hosts and Diseases
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• v.51 no.4
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• pp.449-452
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• 2013

Taenia pisiformis is one of the most important parasites of canines and rabbits. T. pisiformis cysticercus (the larval stage) causes severe damage to rabbit breeding, which results in huge economic losses. In this study, the genetic variation of T. pisiformis was determined in Sichuan Province, China. Fragments of the mitochondrial cytochrome b (cytb) (922 bp) gene were amplified in 53 isolates from 8 regions of T. pisiformis. Overall, 12 haplotypes were found in these 53 cytb sequences. Molecular genetic variations showed 98.4% genetic variation derived from intra-region. $F_{ST}$ and Nm values suggested that 53 isolates were not genetically differentiated and had low levels of genetic diversity. Neutrality indices of the cytb sequences showed the evolution of T. pisiformis followed a neutral mode. Phylogenetic analysis revealed no correlation between phylogeny and geographic distribution. These findings indicate that 53 isolates of T. pisiformis keep a low genetic variation, which provide useful knowledge for monitoring changes in parasite populations for future control strategies.

• Korean Journal of Veterinary Research
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• v.52 no.4
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• pp.239-252
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• 2012

The circulation and infection of avian influenza virus (AIV) in zoos and backyard flocks has not been systematically investigated. In the present study, we surveyed the birds including those in live bird markets (LBMs) and evaluated co-circulation of AIVs among them. Overall, 26 H9N2 AIVs and one H6N2 AIV were isolated from backyard flocks and LBMs, but no AIVs were isolated from zoo birds. Genetic analysis of the HA and NA genes indicated that most of the H9N2 AIVs showed higher similarities to AIVs circulating in domestic poultry than to those in wild birds, while the H6N2 AIV isolate from an LBM did to AIVs circulating in migratory wild birds. In serological tests, 15% (391/2619) of the collected sera tested positive for AIVs by competitive-ELISA. Among them, 34% (131/391) of the sera tested positive for AIV H9 antigen by HI test, but only one zoo sample was H9 positive. Although AIVs were not isolated from zoo birds, the serological results indicated that infection of AIVs might occur in zoos. It was also confirmed that H9N2 AIVs continue to circulate and evolve between backyard flocks and LBMs. Therefore, continuous surveillance and monitoring of these flocks should be conducted to control further epidemics.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.63 no.1
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• pp.1-7
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• 2018

Information on the patterns of genetic diversity and population structure is essential for the rational use and efficient management of germplasms; accurate information aids in monitoring germplasms, and can also be used to predict potential genetic gains. In this study, we assessed genetic diversity, focusing on Korean rice accessions for theand their sustainable conserved diversity. Using DNA profiling with 12 simple sequence repeat (SSR) markers, we detected a total of 333 alleles among 2,016 accessions. The number of alleles ranged from 21 to 53, with an average of 27.8. Average polymorphism information content was 0.797, with the lowest being 0.667 and the highest 0.940. CA cluster analysis and the model-based population structure revealed two main groups that could be subdivided into five subgroups. Analysis of the molecular variance study based on the SSR profile data showed 5% variance among the profiles, whereas we recorded 93% variance among individuals and 2% variance within individuals. Specifically, the utilized diversity for of the breeding program is restricted in that cultivars were located in limited clades. These results revealed that preserving the diversity of Korean landraces could be useful sources for breeding new rice cultivars, and cwould be the basis for the sustainable conservation and utilization of a Korean rice germplasm.

• Journal of Microbiology and Biotechnology
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• v.24 no.2
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• pp.270-279
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• 2014

Microbial community shifts, associated with performance data, were investigated in an anaerobic batch digester treating high-solid food waste under mesophilic conditions using, a combination of molecular techniques and chemical analysis methods. The batch process was successfully operated with an organic removal efficiency of 44.5% associated with a biogas yield of 0.82 L/g $VS_{removal}$. Microbial community structures were examined by denaturing gel gradient electrophoresis. Clostridium and Symbiobacterium organisms were suggested to be mainly responsible for the organic matter catabolism in hydrolysis and acidogenesis reactions. The dynamics of archaeal and methanogenic populations were monitored using real-time PCR targeting 16S rRNA genes. Methanosarcina was the predominant methanogen, suggesting that the methanogenesis took place mainly via an aceticlastic pathway. Hydrogenotrophic methanogens were also supported in high-solid anaerobic digestion of food waste through syntrophism with syntrophic bacterium. Microbial community shifts showed good agreement with the performance parameters in anaerobic digestion, implying the possibility of diagnosing a high-solid anaerobic digestion process by monitoring microbial community shifts. On the other hand, the batch results could be relevant to the start-up period of a continuous system and could also provide useful information to set up a continuous operation.

• Analytical Science and Technology
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• v.13 no.3
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• pp.385-393
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• 2000

A gas chromatography-mass spectrometric (GC/MS) procedure for the determination of amineptine (dihydro-10, 11-dibenzo[a, d] cycloheptenyl-5-amino-7-heptanoic acid) and its main metabolites in human urine was described. Amineptine has been known to be extensively metabolized by the ${\beta}$-oxidation of the heptanoic side chain with formation of pentanoic side chain metabolite ($C_5$-metabolite), and lactamizarion by internal dehydration of (${\beta}$-oxidized metabolite (${\delta}$-lactam). The detection of these compounds was based on acid hydrolysis, liquid-liquid extraction and trimethylsilylated derivatization of the carboxylic acid group. For the determination of amineptine and its metabolites in biological fluids, selected ions at the m/ 192, molecular ion and one of the characteristic ions were monitored by GC/MS. On the excretion study of amineptine in human urine, 70-90% of amineptine, ${\delta}$-lactam, and $C_5$-metabolite were found to be excreted within 4 hours and their excretion completed within 20 hours.

• Journal of Multimedia Information System
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• v.6 no.4
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• pp.293-302
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• 2019

In the past, there was a theory that influenza wasn't transmitted directly from birds but was infected to humans via swains. Recently, molecular level research has progressed, and it was confirmed that the avian influenza virus can directly infected to human lung and intestinal epithelial cells. Three pandemicsin the past 100 years were also infected to humans directly from birds. In view of such scientific background, we are developing a method for screening sick birds by monitoring the physiological characteristics of birds in a contactless manner with sensors. Here, the movement of respiratory muscles and abdominal muscles under autonomic innervation was monitored using a magnet and Hall sensor sewn on the thoracic wall, and other multimedia devices. This paper presents and discusses the results of experiments involving continuous periodic noise discovered during flight experiments with a data logger mounted on a Japanese pheasant from 2012 to 2015. A brief summary is given as the below: 1. Magnet and Hall sensor sewn to the left and right chest walls, bipolar electrocardiograms between the thoracic walls, posterior thoracic air sac pressure, angular velocity sensors sewn on the back and hips, and optical reflection of LEDs (blue and green) from the skin of the hips allow observation of periodic vibrations(fasciculations) in the waves. No such analysis has been reported before. 2. These fasciculations are presumed to be derived from muscle to maintain and control air sac pressure. 3. Since each muscle fiber is spatially Gaussian distributed from the sympathetic nerve, the envelope is assumed to plot a Gaussian curve. 4. Since avian trunk muscles contract periodically at all time, we assume that the sympathetic nerve dominates in their control. 5. The technique of sewing a magnet to the thoracic wall and measuring the strength of the magnetic field with a Hall sensor can be applied to screen for early stage of avian influenza, with a sensor attached to the chicken enclosure.

• The Korean Journal of Mycology
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• v.44 no.1
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• pp.8-15
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• 2016

Fungal contamination is a detrimental factor affecting sawdust media-based shiitake cultivation in greenhouses. During fungal monitoring of greenhouses used for shiitake cultivation, eight fungal species were isolated and identified from indoor air and mushroom flies collected in the greenhouses. The current study reported five species as new in Korea, viz. Ascochyta hordei, Discosia artocreas, Mucor nidicola, Perenniporia medulla-panis, and Pseudozyma prolifica, and confirmed two species, Penicillium charlesii and Penicillium brevicompactum, which were previously recorded in Korea without molecular taxonomic validation. The morphological characteristics and phylogenetic relationships based on nucleotide sequences of the internal transcribed spacer rDNA region or calmodulin gene were described for all identified species.