Immunohistochemistry and polymerase chain reaction for the detection of Lawsonia intracellularis in porcine intestinal tissues with proliferative enteropathy

Detection method of Lawsonia intracellularis was studied in formalin-fixed paraffin-embedded intestinal tissues from 5 naturally infected pigs by immunohistochemistry with a monoclonal antibody against outer membrane protein of L. intracellularis. Warthin-Starry silver stain revealed clusters of argyrophilic, slightly curved rod-shaped organisms in the apical cytoplasm of enterocytes. Immunohistochemical staining with a L. intracellularis-specific monoclonal antibody confirmed the presence of the organism in the apical cytoplasm of hyperplastic enterocytes. The presence of L. intracellularis in the ileum of pig with proliferative enteropathy was confirmed by polymerase chain reaction (PCR) further on the basis of amplification of 319 base pair products specific for porcine L. intracellularis chromosomal DNA. Immunohistochemistry and PCR may be a complementary method to confirm the diagnosis of L. intracellularis infection in pigs.     

Effects of chilled drinking water on the performance of lactating sows and their litters during high ambient temperatures under farm conditions

Heat stress has detrimental effects on lactating sows and their litters, including decreased feed intake and milk production in sows and decreased weight gain in their offspring. It also increases respiration rate, rectal temperature, and weaning to estrus interval in sows. This study was conducted to investigate the effects of chilled water (CW) on the performance of lactating sows and their litters exposed to high ambient temperatures for 21 days. Ninety multiparous sows (Yorkshire × Landrace; parity range: 2 to 5) and their litters (Yorkshire × Landrace × Duroc) were divided into 3 blocks (30 each). Sows and their litters within each block were randomly assigned to one of the three treatment groups (n = 10 each). Each group was provided with drinking water at 10, 15, or 22 °C (control) under farm conditions where ambient temperature above 25 °C was consistent during the experimental period. Sows on CW (both 10 and 15 °C) consumed more feed and water than did control (P < 0.01) and higher estimated milk production compared with control (P < 0.01), but had lower rectal temperature and respiration rate compared with control (P < 0.01). Mean weaning weight (P < 0.01) and average daily gain (P < 0.01) of litters in CW groups were higher than those in control group. There were no differences in the measured variables, with the exception of the respiration rates, between the two CW treatment groups. These results suggest that CW may improve the performance of sows and their litters exposed to high ambient temperatures and the 15 °C water, from an energy viewpoint, may be more effective.     

Effect of soil water stress on the development of stone cells in pear (Pyrus pyrifolia cv. ‘Niitaka’) flesh

This study was carried out to investigate the cause of stone cell formation in pear (Pyrus pyrifolia cv. ‘Niitaka’) flesh. Potted plants grown in a glass house were subjected to water stress conditions without irrigation for 30 days from 30 days before full bloom (BFB treatment), full bloom (FB treatment) and 30 days after full bloom (AFB treatment). Control plants were drip-irrigated daily maintaining a soil matrix potential around −40 ± 5 kPa. The formation of stone cells in pear flesh increased in the FB treatment and AFB treatment plants and this tendency was sustained until the harvest season. Root activity was investigated 60 days after full bloom (DAFB) and the triphenyltetrazolium chloride (TTC) reduction potential, the formazan content and leaf water potential were investigated 30, 45, and 60 DAFB. Root activity decreased progressively due to the effect of water stress. Also, the Ca content in leaf and flesh was lower. The peroxidase activity was high in the flesh at the early stages of fruit growth and decreased at the late stages of fruit growth, and then a higher increase of peroxidase activity was observed in water-stressed fruit. The reduction in calcium content of leaf and fruit in plants under water stress may be related to the reduction of root activity and leaf water potential. The increase in peroxidase activity under water stress may be due to limited calcium absorption. Higher peroxidase activity may induce the accumulation of lignin in the cell wall and promote the formation of stone cells in pear flesh. We conclude that water stress condition during the early stages of fruit growth is one of several factors that determine the formation of stone cells in pear flesh.     

Anti‐microbial Susceptibility for east1+Escherichia coli Isolated from Diarrheic Pigs in Korea

The in vitro susceptibilities of 128 isolates of east1⁺Escherichia coli from pre‐weaned and post‐weaned pigs with diarrhoea were tested with nine commonly used anti‐microbial agents by an agar dilution minimal inhibitory concentration (MIC) procedure according to National Committee for Clinical Laboratory Standards guidelines. For the isolates from pre‐weaned and post‐weaned pigs, most of them were susceptible to low concentrations (MIC₉₀) of tetracycline (4 and 2 μg/ml), ceftiofur (2 and 2 μg/ml), and colistin (4 and 2 μg/ml). Marked resistance was found in others.     

Epizootiologic survey for Babesia microti among small wild mammals in northeastern Eurasia and a geographic diversity in the beta-tubulin gene sequences

We previously reported that small wild rodents in Japan harbor two types of novel Babesia microti-like parasites (designated as Hobetsu and Kobe types), but not the type commonly found in the northeastern United States (U.S. type) where human babesiosis is endemic. To determine whether these new types of parasites are distributed in places surrounding Japan, an epizootiologic survey was undertaken in three geographically distant areas in northeastern Eurasia; South Korea, Vladivostok in Russia, and Xinjiang in China. Blood samples were collected from a total of 387 animals comprising 24 species. DNAs extracted from the samples were tested by nested PCR targeting babesial nuclear small-subunit rRNA gene (rDNA), which revealed that small rodents harboring B. microti exist in all three survey areas. Sequence analysis showed that all PCR-positive samples had rDNA sequences virtually identical to that of U.S.-type B. microti. However, when beta-tubulin gene sequences were compared, evident geographic variations were seen. By use of primers specific for each of the beta-tubulin genes of Kobe-, Hobetsu-, and U.S.-type parasites, a type-specific PCR was developed. Parasite with Hobetsu- or Kobe-type sequence was not detected from any of the three survey areas. These findings suggest that U.S.-type B. microti is widely distributed among small wild mammals in temperate zones of not only North America, but also Eurasia, whereas that Hobetsu- and Kobe-type parasites may be uniquely distributed in Japan.     

Isolation and characterization of antimicrobial-resistant Escherichia coli from national horse racetracks and private horse-riding courses in Korea

Limited information is available regarding horse-associated antimicrobial resistant (AR) Escherichia (E.) coli. This study was designed to evaluate the frequency and characterize the pattern of AR E. coli from healthy horse-associated samples. A total of 143 E. coli (4.6%) were isolated from 3,078 samples collected from three national racetracks and 14 private horse-riding courses in Korea. Thirty of the E. coli isolates (21%) showed antimicrobial resistance to at least one antimicrobial agent, and four of the AR E. coli (13.3%) were defined as multi-drug resistance. Most of the AR E. coli harbored AR genes corresponding to their antimicrobial resistance phenotypes. Four of the AR E. coli carried class 1 integrase gene (intI1), a gene associated with multi-drug resistance. Pulsed-field gel electrophoretic analysis showed no genetic relatedness among AR E. coli isolated from different facilities; however, cross-transmissions between horses or horses and environments were detected in two facilities. Although cross-transmission of AR E. coli in horses and their environments was generally low, our study suggests a risk of transmission of AR bacteria between horses and humans. Further studies are needed to evaluate the risk of possible transmission of horse-associated AR bacteria to human communities through horse riders and horse-care workers.     

Immunohistochemical detection of scrapie prion proteins in clinically normal sheep in Pennsylvania.

Following diagnosis of scrapie in a clinically suspect Suffolk sheep, 7 clinically normal flockmates were purchased by the Pennsylvania Department of Agriculture to determine their scrapie status using an immunohistochemical procedure. Two of the 7 euthanized healthy sheep had positive immunohistochemical staining of the prion protein of scrapie (PrP-Sc) in their brains, nictitating membranes, and tonsils. The PrP-Sc was localized in the areas of the brain where, histopathologically, there was neurodegeneration and astrocytosis. The PrP-Sc occurred within germinal centers of the affected nictitating membranes and tonsils and was located in the cytoplasm of the dendrite-like cells, lymphoid cells, and macrophages. These results confirm that immunohistochemical examination of the nictitating membrane can be used as a screen for the presence of scrapie infection in clinically normal sheep at a capable veterinary diagnostic laboratory. In sheep with a PrP-Sc-positive nictitating membrane, the diagnosis of scrapie should be confirmed by histopathology and immunohistochemical examination of the brain following necropsy. Following full validation, immunohistochemistry assays for detection of PrP-Sc in nictitating membrane lymphoid tissues can improve the effectiveness of the scrapie control and eradication program by allowing diagnosis of the disease in sheep before the appearance of clinical signs.