• Parasites, Hosts and Diseases
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• v.56 no.6
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• pp.559-565
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• 2018

The identification and characterization of pathogenic and zoonotic tick-borne diseases like granulocytic anaplasmosis are essential for developing effective control programs. The differential diagnosis of pathogenic Anaplasma phagocytophilum and non-pathogenic A. phagocytophilum-like Anaplasma spp. is important for implementing effective treatment from control programs. The objective of the present study was to investigate the prevalence of Anaplasma spp. in horses in Korea by nucleotide sequencing and restriction enzyme fragment length polymorphism assay. Of the 627 horses included in the study, only 1 (0.2%) was infected with A. phagocytophilum. Co-infection with A. phagocytophilumlike Anaplasma spp. was not detected in the study. The 16S rRNA sequence of A. phagocytophilum was similar (99.5-100%) to A. phagocytophilum 16S rRNA isolated from horses in other countries. PCR adapted to amplify A. phagocytophilum groEL and msp2 genes failed to generate amplicons, suggesting genetic diversity in these genes. This study is the first molecular detection of A. phagocytophilum in horses in Korea. Human granulocytic anaplasmosis and animal infection of A. phagocytophilum have been reported in Korea recently. Because of vector tick distribution, global warming, and the increase of the horse industry, horses should be considered as a potential reservoir for A. phagocytophilum, and cross infectivity should be evaluated even though a low prevalence of infection was detected in this study. Furthermore, continuous surveillance and effective control measures for A. phagocytophilum should be established to prevent disease distribution and possible transmission to humans.

• Korean Journal of Veterinary Service
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• v.45 no.1
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• pp.1-11
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• 2022

A novel porcine circovirus 4 (PCV4) was recently identified in Chinese and Korean pig herds. Although several conventional polymerase chain reaction (cPCR) and real-time PCR (qPCR) assays were used for PCV4 detection, more sensitive and reliable qPCR assay is needed that can simultaneously detect PCV4 and internal positive control (IPC) to avoid false-negative results. In the present study, a duplex qPCR (dqPCR) assay was developed using primers/probe sets targeting the PCV4 Cap gene and pig (glyceraldehyde-3-phosphate dehydrogenase) GAPDH gene as an IPC. The developed dqPCR assay was specifically detected PCV4 but not other PCVs and porcine pathogens, indicating that the newly designed primers/probe set is specific to the PCV4 Cap gene. Furthermore, GAPDH was stably amplified by the dqPCR in all tested viral and clinical samples containing pig cellular materials, indicating the high reliability of the dqPCR assay. The limit of detection of the assay 5 copies of the target PCV4 genes, but the sensitivity of the assay was higher than that of the previously described assays. The assay demonstrated high repeatability and reproducibility, with coefficients of intra-assay and inter-assay variation of less than 1.0%. Clinical evaluation using 102 diseased pig samples from 18 pig farms showed that PCV4 circulated in the Korean pig population. The detection rate of PCV4 obtained using the newly developed dqPCR was 26.5% (27/102), which was higher than that obtained using the previously described cPCR and TaqMan probe-based qPCR and similar to that obtained using the previously described SYBR Green-based qPCR. The dqPCR assay with IPC is highly specific, sensitive, and reliable for detecting PCV4 from clinical samples, and it will be useful for etiological diagnosis, epidemiological study, and control of the PCV4 infections.

• Reproductive and Developmental Biology
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• v.35 no.4
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• pp.479-483
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• 2011

The objective of this study was to determine the effects of E. coli isolated from porcine semen on sperm viability, motility, and semen pH. Semen samples were prepared using commercial extender, $Seminark^{Pro}$ (Noahbio Tech, Korea) that did not contain antibiotics. And 4 different levels of E. coli were artificially innoculated to semen with following concentrations; 4,000 of sperms with 1 of E. coli (T1), 400 with 1 (T2), 40 with 1 (T3), and 4 with 1 (T4). Semen samples were preserved at $17^{\circ}C$ for 5 days in semen storage box until analyzed by flowcytometer. Aliquots were subjected to measure the sperm viability (Live/$Dead^{(R)}$ stain), motility (mitochondrial function), and semen acidity (pH) from day 0 (day of semen collection) to day 5. Sperm motility and viability were significantly decreased (p<0.05) on day 0 (4 hrs after preservation at $17^{\circ}C$) in T3 and T4 compared to control groups and were significantly decreased (p<0.05) in all groups from day 3. Sample pH was acidic in T3 (6.90~6.86) and T4 (6.86~6.65) from day 3 to day 5 (p<0.05). On the other hand, sample pH was maintained 7.0~7.1 in control, T1, and T2 during the experimental period. Sperm motility and viability were significantly decreased from day 0 to day 5 compared to control in samples contaminated with E. coli above a value of 40:1 ($20{\times}10^6$ sperm cells/ml : $5{\times}10^5$ cfu/ml). Even on day 1 in T4 and on day 3 in T3, semen pH was acidic probably due to the acidification of dead spermatozoa. These results suggest that E. coli contamination has a concentration-dependent detrimental effect on extended porcine semen quality.

• Korean Journal of Veterinary Service
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• v.39 no.4
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• pp.211-219
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• 2016

For determining the prevalence of major enteric pathogens, clinical examination and etiological diagnosis were carried out on 75 Korean pig farms. Enteric disease-suspected signs were observed in 90.7% of the farms and the incidence and severity were higher in younger age groups of the pigs. Five of seven pathogens were detected in 375 fecal samples collected from the 75 farms, and the farm-level prevalence of porcine rotavirus group A (PoRVA), pathogenic Escherichia (E.) coli, Lawsonia (L.) intracelluraris, Salmonella spp., and Brachyspira (B.) hyodysenteriae was 54.7%, 54.7%, 16.0%, 10.7% and 2.7%, respectively. PoRVA was extensively infected in suckling and weaning pig groups. The prevalence of pathogenic E. coli was highest in suckling period, and after the period, it exhibited a tendency to decrease. Salmonella spp. and L. intracelluraris were detected in all feeding groups of pigs in a ratio of 1.3~6.7%. B. hyodysenteriae was detected in 1.3~2.7% of growing and fattening pig groups but not detected in suckling and weaning pig groups. At least one or more pathogens were detected in 30.1% of 375 fecal samples. Among these, 25.0% or 5.1% of cases were single or mixed infection. Enteric disease signs of the pigs were significantly co-related with the detection of PoRVA, pathogenic E. coli or Salmonella spp. (P<0.01) but not with L. intracelluraris or B. hyodysenteriae (P>0.05). Conclusively, it will be expected that these data obtained in this study are very useful for subsequent studies and prevention strategies for swine enteric disease in Korean pig farms.

• Journal of Life Science
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• v.28 no.5
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• pp.555-562
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• 2018

Swine influenza virus (SIV) causes one of the most common diseases of the pig population, and its subtypes are determined by hemagglutinin (HA) and neuraminidase (NA). Recently, the SIV subtype diagnosis has been developed. The method using antigen-antibody reaction rather than PCR was mainly used because of the large change in the ribonucleotide sequences of SIV. Here, we have developed 10 diagnostic primer sets through multi-nucleotide sequences alignment of spreaded SIV since 2008 in Korea and then optimized the reaction of the one-step RT-PCR for rapid determination of SIV subtype. In addition, specific primers were designed to early determine the pandemic SIV by detecting unique M sequences proven in highly infectious and virulent subtypes of the influenza H1N1 (pH1N1). Here, some of the SIVs spread in Korea from 2008 to 2014 have been tested to determine the subtypes and pandemic potential of SIV. All diagnostic primer sets were found to be able to accurately determine the SIV subtype and to detect the pandemic SIV. In conclusion, it was confirmed that the optimized one-step RT-PCR analysis using these primer sets is useful for rapid diagnosis of SIV subtypes. These results can be used for development of SIV subtype diagnostic kit to early detect before virulent SIV spreads do.

• Research in Plant Disease
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• v.20 no.4
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• pp.275-282
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• 2014

The occurrence of post-harvest diseases and their pathogens in 'Campbell Early' which is the most produced grape in Korea was investigated. The 'Campbell Early' grapes produced in 3 main grape-producing areas were stored in a cold room ($0-4^{\circ}C$) for 2 weeks then at room temperature for 4 weeks prior to investigation. The major post-harvest diseases occurred were gray mold, blue mold, ripe rot, new decay 1, and new decay 2. Pathogens isolated from the symptoms were identified as Botrytis cinerea for gray mold, Penicillium sclerotiorum for blue mold and Collectrichum acutatum for ripe rot. Pathogens for new decay 1 and new decay 2 were not identified yet. Incidences of new decay 1 and new decay 2 were much higher than the other 3 decays in all grapes produced from 3 areas. Gray mold and blue mold occurred at much lower frequencies than these two decays, and ripe rot occurred least.

• Parasites, Hosts and Diseases
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• v.56 no.5
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• pp.437-446
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• 2018

To investigate the prevalence of Toxoplasma gondii in pork on the market in Korea, an in-house enzyme-linked immunosorbent assay for tissue fluid (CAU-tf-ELISA) was developed using a soluble extract of T. gondii RH strain tachyzoites. As the standard positive controls, the piglets were experimentally infected with T. gondii: Group A (1,000 cysts-containing bradyzoites), Group B (500 cysts-containing bradyzoites) and Group C ($1.0{\times}10^3$ or $1.0{\times}10^4$ tachyzoites). The CAU-tf-ELISA demonstrated infection intensity-dependent positivity toward tissue fluids with average cut-off value 0.15: 100% for Group A, 93.8% for Group B and 40.6% for Group C. When tissue-specific cut-off values 0.066-0.199 were applied, CAU-tf-ELISA showed 96.7% sensitivity, 100% specificity, 100% positive and 90.0% negative predictive values. When compared with the same tissue fluids, performance of CAU-tf-ELISA was better than that of a commercial ELISA kit. Of the 583 Korea domestic pork samples tested, anti-T. gondii antibodies were detected from 9.1% of whole samples and 37.9% from skirt meat highest among pork parts. In the 386 imported frozen pork samples, 1.8% (skirt meat and shoulder blade) were positive for anti-T. gondii antibodies. In Korea, prevalence of anti-T. gondii antibodies in the pork on retail markets appeared high, suggesting that regulations on pig farming and facilities are necessary to supply safe pork on the tables.

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.7
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• pp.1044-1049
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• 2014

This study was conducted in order to investigate whether the presence of light or different colors of light would influence the energy expenditure and behavior of broiler chickens. Eight 8-week-old broiler chickens were adapted to a respiration chamber (Length, 28.5 cm; Height, 38.5 cm; Width, 44.0 cm) for one week prior to the initiation of the experiment. In experiment 1, energy expenditure and behavior of the chickens were analyzed in the presence or absence of light for four days. Chickens were exposed to 6 cycles of 2 h light/2 h dark period per day. In experiment 2, the broiler chickens that had been used in experiment 1 were used to evaluate the effect of 4 different wavelength light-emitting diodes (LEDs) on the energy expenditure and behavior of broiler chickens. The LEDs used in this study had the following wavelength bands; white (control), red (618 to 635 nm), green (515 to 530 nm) and blue (450 to 470 nm). The chickens were randomly exposed to a 2-h LED light in a random and sequential order per day for 3 days. Oxygen consumption and carbon dioxide production of the chickens were recorded using an open-circuit calorimeter system, and energy expenditure was calculated based on the collected data. The behavior of the chickens was analyzed based on following categories i.e., resting, standing, and pecking, and closed-circuit television was used to record these behavioral postures. The analysis of data from experiment 1 showed that the energy expenditure was higher (p<0.001) in chickens under light condition compared with those under dark condition. The chickens spent more time with pecking during a light period, but they frequently exhibited resting during a dark period. Experiment 2 showed that there was no significant difference in terms of energy expenditure and behavior based on the color of light (white, red, green, and blue) to which the chickens were exposed. In conclusion, the energy expenditure and behavior of broiler chickens were found to be strongly affected by the presence of light. On the other hand, there was no discernible difference in their energy expenditure and behavior of broiler chickens exposed to the different LED lights.

• Reproductive and Developmental Biology
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• v.37 no.4
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• pp.263-267
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• 2013

Pig producers have been shown keen interest of the number of spermatozoa in a semen dose since pig artificial insemination introduce. However, determining the minimal number of spermatozoa need per AI without detrimental effect on overall reproductive performances is not an easy question to answer. To increase the efficiency of semen utilization in pig AI, optimum number of spermatozoa per dose needed to determine. The objective of this study was to determine the reproductive performance and factors that affect on-farm application of low-dose semen insemination in sows. Data were collected from Darby Genetics AI studs from 4th of June to 7th of July, 2012 (n=401). The numbers of parturition were 84, 234 and 83 in sows inseminated with doses of $1.5{\times}10^9$, $2.0{\times}10^9$ and $2.5{\times}10^9$ spermatozoa in 100ml extender, respectively. There were no significant differences on reproductive performances such as gestation period, total born, total born alive, stillbirth and mummy in sows inseminated with different semen doses. The average number of born alive was 10.5, 11.0 and 10.4 from sows inseminated with $1.5{\times}10^9$, $2.0{\times}10^9$ and $2.5{\times}10^9$ sperms, respectively. Also, number of spermatozoa per dose did not affect litter size (p>0.10). There were no significant differences of maternal genetic line difference on gestation period, total number born, number born alive, born dead and mummy. The estimated correlation coefficients of the different semen doses with total number born, number born alive, born dead and mummy were r=-0.00, -0.01, 0.02 and 0.02, respectively. Taken together, the result of this study suggested that when semen was appropriately inseminated after induced ovulation, insemination with low-dose ($1.5{\sim}2.0{\times}10^9$) semen dose not adversely affect sow's fertility.

• Research in Plant Disease
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• v.22 no.3
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• pp.158-167
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• 2016

A pathogen that causes a new disease on green pumpkin in the nursery and the field was characterized and identified. Symptoms of the disease on green pumpkin were water soaking lesions and spots with strong yellow halo on leaf, brown lesion on flower, and yellow spot on fruit. The bacterial isolates from the leaf spot were pathogenic on the 8 curcubitaceae crop plants, green pumpkin, figleaf gourd, wax gourd, young pumpkin, zucchini, cucumber, melon, and oriental melon, whereas they did not cause the disease on sweet pumpkin and watermelon. They were Gram-negative, rod shape with polar flagella, fluorescent on King's B agar and LOPAT group 1a by LOPAT test. Their Biolog substrate utilization patterns were similar to Pseudomonas syringae pv. syringae's in Biolog database. Phylogenetic trees with 16S rRNA gene sequences and multilocus sequence typing (MLST) with nucleotide sequences of 4 housekeeping genes, gapA, gltA, gyrB, rpoD and those of P. syringae complex strains in the Plant Associated and Environmental Microbes Database (PAMDB) showed that the green pumpkin isolates formed in the same clade with P. syringae pv. syringae strains. The clade in MLST tree was in the genomospecies 1 group. The phenotypic and genotypic characteristics suggested that the isolates from green pumpkin lesion were P. syringae pv. syringae.