Genotypic prevalence of the adhesin involved in diffuse adherence in Escherichia coli isolates in pre-weaned pigs with diarrhoea in Korea

A total of 1002 Escherichia coli strains isolated from pre-weaned pigs with diarrhoea on 1114 swine farms were screened for the presence of the adhesin involved in diffuse adherence (AIDA) gene by polymerase chain reaction (PCR). Escherichia coli isolates that carried AIDA genes were also tested by PCR for the detection of five fimbriae (F4, F5, F6, F18 and F41), heat-stable (STa, STb) and heat-labile (LT) enterotoxin, enteroaggregative E. coli heat-stable enterotoxin 1 (EAST1), and Shiga toxin 2 oedema disease (Stx2e) genes. Twenty-three (2.3%) of the 1002 E. coli isolates carried the gene for AIDA. Among 23 isolates shown to carry genes for AIDA, three carried the AIDA gene as the only shown virulence factor. Other isolates carried other virulence factor genes in addition to AIDA. Four isolates carried genes for at least one of the fimbrial adhesins and enterotoxins. Sixteen isolates carried genes for enterotoxins only. The AIDA may represent an additional virulence determinant in pre-weaned pigs with diarrhoea.     

Geographical comparison of genetic diversity in Asian landrace wheat (<Emphasis Type="Italic">Triticum aestivum</Emphasis> L.) germplasm based on high-molecular-weight glutenin subunits

Glutenin largely determines wheat bread baking quality. As high-molecular-weight glutenin subunit (HMW-GS), related to Glu-1 loci, determines wheat flour elasticity, it correlates strongly with bread-making quality. This study was aimed at clarifying genetic variations in bread-making characteristics between East and West Asian wheat landrace germplasms, by investigating HMW-GS allelic composition of 1068 wheat accessions. Herein, the accession number having reported HMW-GS pattern in previous studies was 855. However, the accession number with newly detected HMW-GS patterns was 114. These new HMW-GS patterns were classified into 4 types based on similarity. Eight Korean accessions with these four types were identified. Concerning landrace germplasm nature, 99 accessions showed heterogeneous patterns caused by seed mixture. The Glu-1 loci allelic variation analysis, revealed that the percentages of Glu-A1c (73.6%), Glu-B1b (60.2%), and Glu-D1a (68.5%) were highest at Glu-A1, Glu-B1, and Glu-D1 loci, respectively. The incidence of preferable alleles for bread baking was high in Chinese accessions. In bread-making quality evaluation using Glu-1 score, 24 among 35 accessions with full score were from China. The polymorphic information content index of each origin based on HMW glutenin subunit combination showed that West Asian and neighboring-regional landraces, excluding Afghanistan ones, were more diverse than East Asian landraces excluding Chinese ones. Cluster analysis based on Glu-1 allelic combination showed that many Korean, Japanese, and Afghan accessions were in the same group. However, many Chinese and other West Asian accessions were in the other group despite geographical distance.     

Pharmacokinetic study of meropenem in healthy beagle dogs receiving intermittent hemodialysis

Meropenem, a second carbapenem antimicrobial agent with a broad spectrum of activity, is used to treat sepsis and resistant‐bacterial infections in veterinary medicine. The objective of this study was to identify the pharmacokinetics of meropenem in dogs receiving intermittent hemodialysis (IHD) and to determine the proper dosing in renal failure patients receiving IHD. Five healthy beagle dogs were given a single i.v. dose of 24 mg/kg of meropenem and received IHD. The blood flow rate, dialysate flow, and ultrafiltration rate were maintained at 40 mL/min, 300 mL/min, and 40 mL/h, respectively. Blood samples were collected for 24 h from the jugular vein and from the extracorporeal arterial and venous line. Urine samples and dialysate were also collected. The concentrations of meropenem were assayed using HPLC/MS/MS determination. The peak plasma concentration was 116 ± 37 μg/mL at 15 min. The systemic clearance was 347 ± 117 mL/h/kg, and the steady‐state volume of distribution was 223 ± 67 mL/kg. Dialysis clearance was 71.1 ± 34.3 mL/h/kg, and the extraction ratio by hemodialysis was 0.455 ± 0.150. The half‐life (T_1/2) in dogs with IHD decreased compared with those without IHD, and the reduction in T_1/2 was greater in renal failure patients than in normal patients. Sixty‐nine percent and 21% of the administered drug were recovered by urine and dialysate in the unchanged form, respectively. In conclusion, additional dosing of 24 mg/kg of meropenem after dialysis could be necessary according to the residual renal function of the patient based on the simulated data.     

The grafting of recycled polyol from waste polyurethane foam onto new polyurethane and its impact on shape recovery and water vapor permeation

Recycled polyols from waste polyurethane (PU) foams were grafted onto PU to improve the properties such as tensile strength, shape recovery, low-temperature flexibility, and water compatibility. The recycled polyol was either purified by column chromatography before grafting or was used directly for grafting. The soft segment melting temperature of PU did not notably increase with the addition of polyol, whereas the glass transition temperature increased with increased polyol content. The tensile strength sharply increased at low polyol content and decreased at high polyol content, while the strain at break did not significantly change with an increase in polyol content. The shape recovery at 10 oC notably improved compared with unmodified PU and remained high after four cyclic tests. Polyol-grafted PU demonstrated better lowtemperature flexibility and reduced the water vapor permeability of PU membranes. Overall, grafting recycled polyol onto PU significantly improved the tensile stress, shape recovery, and low-temperature flexibility of PU.     

Identification of a RAPD marker linked to a brown planthopper resistance gene in rice

We report the tagging of a brown planthopper (BPH) resistance gene (Bph–1) in rice using RAPD and RFLP markers. The Korean rice variety ‘Gayabyeo’ has dominant duplicate genes including Bph–1 conferring resistance to biotype 1 of BPH. Bulked segregant RAPD analysis was employed for rapid identification of DNA markers linked to resistance genes. For tagging these two genes, an F2F3 population from a ‘Gayabyeo’ × ‘Nagdongbyeo’ cross was developed and evaluated for BPH resistance. Three bulked DNAs from two groups of homozygous BPH resistant (each for Bph–1 and the other unknown gene) and homozygous susceptible F2 plants were analyzed by RAPD using 140 random oligomers. One primer, OPD–7 yielded a 700-bp fragment that was present in Gayabyeo and resistant F2 plants (homozygous for Bph-1 locus) but absent in Nagdongbyeo and susceptible F2 plants. Cosegregation of this marker with Bph-1 was verified using an F2 population segregating for Bph-1. Chromosomal regions surrounding the Bph-1 were examined with additional RFLP and microsatellite markers on chromosome 12 to define the location of the RAPD marker and Bph-1. Use of this RAPD marker could facilitate early selection of resistant lines for BPH. This revised version was published online in July 2006 with corrections to the Cover Date.     

Determination of Feeder Cell‐Based Cellular Niches Supporting the Colonization and Maintenance of Spermatogonial Stem Cells from Prepubertal Domestic Cat Testes

Recently, isolation and in vitro culture of putative spermatogonial stem cells (SSCs) in the domestic cat have been conducted. However, the cellular niche conditions that facilitate the establishment and long‐term maintenance of feline SSCs (FSSCs) have not been described. Therefore, we investigated the type of feeder cells used to stimulate colony formation and growth of FSSCs among the various factors in the FSSC niche. Spermatogonial stem cells isolated from feline testes were cultured on mitotically inactivated testicular stromal cells (TSCs) derived from cats, dogs and mice, and mouse embryonic fibroblasts (MEFs). The formation and growth of colonies derived from SSCs cultured on each type of feeder cell were identified at passage 0, and the morphology, alkaline phosphatase (AP) activity and expression of SSC‐specific genes in surviving colonies were investigated at passage 4. Among these diverse feeder cells, TSCs from cat showed the greatest colony formation, growth and maintenance of FSSCs, and SSC colonies cultured by passage 4 showed a typical dome‐shaped morphology, AP activity and expression of SSC‐specific genes (NANOG, OCT4, SOX2 and CD9). Accordingly, these results demonstrate that feline TSCs could be used as feeder cells to support the establishment and maintenance of SSCs from domestic cats.     

Interpreting the temperature-induced response of ammonia oxidizing microorganisms in soil using nitrogen isotope fractionation

Purpose  

Although nitrification plays a key role in the fate of soil nitrogen (N) under global warming, little information is available for the nitrifiers’ response to changing temperatures. Nitrogen isotope fractionation associated with nitrification can be a proxy of nitrifiers’ sensitivity to changing temperature. We hypothesized that the temperature-induced balance between the transport of substrate NH₄⁺ into the microbial cell (supply) and the intracellular NH₄⁺ oxidation (consumption) governs the intracellular NH₄⁺ concentration and then affects nitrification rates and associated isotope fractionations. This study was conducted to understand the microbial response of NH₄⁺ oxidation to changing temperatures by examining the effect of changing temperature on nitrification rate and apparent isotope fractionation.     

Factors affecting the consecutive and simultaneous coupling with bis-diazotized benzidine of protein antigen to sheep red blood cells

The dynamics of attaching antigen to sheep red blood cells using bis-diazotized benzidine (BDB) were studied using purified protein derivative tuberculin as antigen. Factors considered were the concentration of antigen and BDB, temperature, time and simultaneous or consecutive coupling. Simultaneous coupling at 42°C for 10 min gave the most specific haemagglutination reactions and was simple to carry out.     

Isolation and characterization of porcine circovirus type-2 from sera of stillborn fetuses

In order to examine an association between porcine circovirus type-2 (PCV2) infection and reproductive failure in pigs, sera (n = 171) from stillborn fetuses were collected from 3 different farms with prolonged histories of reproductive problems. These sera were tested for the presence of antibodies to PCV2 using an immunoperoxidase monolayer assay. Of the 171 sera tested, 28 had PCV2 antibody titers of ≥ 1:16. When these 28 samples were tested by a polymerase chain reaction assay, 13 were found to contain PCV2 viral DNA. Of these 13 samples containing both PCV2 antibodies and viral DNA, 9 yielded PCV2 on virus isolation. Amino acid sequences comprising open reading frame 2 of PCV2 from 2 of these isolates were compared to PCV2 isolates from cases of post-weaning multi-systemic wasting syndrome (PMWS). The amino acid sequences of the 2 isolates from stillborn pigs were shown to be nearly identical to each other, as well as to other PCV2 isolates associated with reproductive failure. When compared with PMWS isolates, the isolates from the stillborn fetuses showed differences of at least 2 amino acids. These results confirm previous findings that transplacental infection of PCV2 occurs in the field and that stillbirths in pigs may be associated with PCV2 infections. At present, the significance of minor differences in amino acid sequences is not known.