Expression of MAP kinases and connexins in the differentiation of rat mammary epithelial cells

Gap junctional intercellular communication (GJIC) is involved in the regulation of many cellular processes. MAP kinases are known to affect GJIC and phosphorylation of connexin (Cx). MAP kinases can also be a regulator of cell proliferation and growth. This study was undertaken to show the relevance between expression patterns of Cxs and MAP kinases in rat mammary epithelial cells (RMECs). In order to characterize the RMECs, they were stained with Peanut lectin, which indicates most alveolar epithelial cells, and Thy-1.1 was used as a marker of luminal epithelial cells or myoepithelial cells, respectively. We studied the expression patterns of major gap junction proteins, Cx26, 32, and 43 in RMECs. Western blot analysis demonstrated that Cx26 gradually decreased from day 2, while Cx32 was expressed constantly from day 1 to 14. Cx43 dramatically increased on day 5 and decreased thereafter. The expression patterns and phosphorylation of ERK1/2 and JNK were similar to Cx43, but expression of p38 was like that of Cx32. These results showed that the MAP kinases that comprise ERK1/2, p38, and JNK were involved in regulation of Cxs. Our data suggests that GJIC plays an important role during rat mammary differentiation and that MAP kinases may be closely related functionally to regulate the gap junction.     

Age-related changes in calretinin-immunoreactive periglomerular cells in the rat main olfactory bulb

The changes of calretinin (CR)-immunoreactive periglomerular cells in the glomerular layer of the main olfactory bulb (MOB) were investigated in rats differing ages from postnatal month 1 (PM 1) to PM 24. The number of cresyl violet-positive periglomerular cells was similar between PM 1 and PM 12, but they decreased slightly in the PM 24 group. The size of CR-immunoreactive periglomerular cells in the glomerular layer increased with age, while their numbers did not change significantly in the PM 6-PM 24 groups. In the PM 24 group, numbers of CR-positive periglomerular cell bodies and their processes decreased, while the size of CR-positive cell bodies in the glomeruli was larger than that of the previous groups. These results suggest that CR-immunoreactive periglomerular cells in the rat MOB are well-developed in the PM 6 group, and that periglomerular cells in the PM 24 group show poor CR-immunoreactivity compared to those in the PM 6 group.     

A multi-microbe probiotic formulation processed at low and high drying temperatures: effects on growth performance,nutrient retention and caecal microbiology of broilers

1. Two experiments were conducted to evaluate a multi-microbe probiotic formulation processed at low (LT) or high (HT) drying temperature.

2. In both the experiments, 640 d-old Ross male chicks were randomly allotted to 4 treatments on the basis of initial BW for 35 d experiments.

3. In experiment one, dietary treatments were a negative control (NC; basal diet without any antimicrobial); positive control (PC; basal diet +10?mg/kg avilamycin); basal diet with 0·3% probiotic LT; and basal diet with 0·3% probiotic HT.

4. Improved overall weight gain, FCR and retention of CP were observed in birds fed the PC and probiotic diets when compared with birds fed the NC diet. At d 21, birds fed the probiotic and NC diets had more caecal Bifidobacterium and total anaerobes than birds fed the PC diet; while birds fed the PC and probiotic diets had fewer caecal Clostridium than birds fed the NC diet at d 35.

5. In experiment two, a 2?×?2 factorial arrangement of treatments was employed to evaluate the effects of two concentrations of probiotic HT (0·30 or 0·60%) and avilamycin (0 or 10?mg/kg).

6. Birds fed the 0·60% probiotic HT diet showed improved overall weight gain and CP retention, higher Lactobacillus and Bifidobacterium in the caecum, and reduced Clostridium and coliforms in the caecum. Inclusion of avilamycin improved the overall weight gain and feed intake, and reduced the caecal Clostridium and Bifidobacterium population.

7. In conclusion, high drying temperature had no effect on the efficacy of the multi-microbe probiotic formulation; while the probiotic HT formulation was more effective at the 0·60% level. Moreover, inclusion of avilamycin improved performance of birds but did not have any interaction with probiotics.     

Comparison of vancomycin-resistant enterococci isolates from human, poultry and pigs in Korea

Vancomycin-resistant enterococci (VRE) have emerged as an important nosocomial pathogen. Since 1989, a rapid increase in the incidence of enterococcal bacteremia and endocarditis by VRE has been reported. The use of avoparcin in animal husbandry is reportedly associated with the appearance of VRE. In this study, a multiplex polymerase chain reaction (PCR) method was established to detect and differentiate resistant types of enterococci, which specifically amplify the four van genes that encode vancomycin resistance elements. Using this method, we investigated the incidence rates and types of VRE from two types of farms: those that had used avoparcin and those that had not used avoparcin. A total of 1091 animal fecal samples were collected from 70 pig farms and 32 poultry farms. A total of 425 enterococci were isolated from the fecal samples. Among the 425 isolates, six showed a pattern of high-level vancomycin resistance (Minimal Inhibitory Concentration, MIC: 64-256 microg/ml). Out of six high-level VRE, three were isolated from poultry farms that had used avoparcin and three were not. The six high-level VRE harbored the vanA gene. Sixty-seven of 425 isolates that showed a pattern of low-level vancomycin resistance (MIC: 4-8 microg/ml) were associated with the presence of vanC-1 or vanC-2/3 gene. We also performed a repetitive extragenic palindromic PCR (rep-PCR) method to compare the genetic relatedness between the high-level VRE of six animal isolates and 31 human isolates. None of the animal isolates had a similar rep-PCR pattern as the human isolates but similarities between human VRE isolates were observed.     

Characterization and differentiation of <Emphasis Type="Italic">Erwinia carotovora</Emphasis> strains from mulberry trees

We recently reported that two diverse types (types 1 and 2) were identified among strains of Erwinia carotovora from mulberry trees. Type 1 strains were similar to E. carotovora subsp. carotovora (Ecc), whereas type 2 strains were distinct from Ecc and other E. carotovora strains. In this study, seven more mulberry strains of type 2 and reference strains were analyzed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and randomly amplified of polymorphic DNA (RAPD). On the basis of SDS-PAGE profiles of whole-cell proteins, type 2 strains had high similarity with one another. In addition, they had an unique peptide band with a molecular mass of approximately 28kDa. RAPD analysis showed that they were also effectively differentiated by a strong, specific RAPD fragment for type 2 strains. Based on these two approaches, we have confirmed that the present type 2 strains from mulberry can be discriminated clearly from other soft rot Erwinia species.     

Biofilm-forming associated genotypic and phenotypic characteristics of Staphylococcus spp. isolated from animals and air

Biological monitoring is performed to detect and analyze microorganisms that have continuously made an effort to survive in the environment. Of such microorganisms, Staphylococcus spp. is considered a common cause of nosocomial and environmental infections., Microbial Surface Components Recognizing Adhesive Matrix Molecules (MSCRAMMs) are required for the adhesion and biofilm formation of Staphylococci. Thirty-six and thirty-five Staphylococci isolated from animals and air, respectively, were analyzed. Biofilm formation and ten MSCRAMM genes were investigated using Congo red agar, tissue culture plate methods, and PCR. Airborne isolates were shown to have higher adherence and stronger biofilm formation than those from animals. The prevalence of MSCRAMM genes from air isolates was also higher than those from animals. Of the genes, eno was mainly associated with biofilm formation in both animals and airborne isolates (P < 0.05). Moreover, the rate of airborne isolates harboring the eno gene was higher than in animal isolates. These results indicated that analysis of MSCRAMM genes with a phenotypic assay might be a helpful bacterial control system for the environment.     

Larval Development and Rearing of Longtooth Grouper Epinephelus bruneus in Jeju Island, Korea

Eggs and sperm were obtained from a female (6.3 kg/BW) and a male (8.4 kg/BW) longtooth grouper Epinephelus bruneus following HCG injection in July 2003. The eggs were fertilized artMcially with the sperm and incubated in one of two 50-m³ tanks after washing the fertilized eggs. The fertilized eggs were 830–950 pn (average 900 ± 2 μm) in diameter and the respective fertilization and hatching rates were 97.7 ± 0.6% and 96.8 ± 0.5% at a water temperature of 25.0 ± 0.5 C. With this regime, the survival rate by day 93 was 7.5% in the 50-m³ tank. The elapsed time from hatching to opening the mouth was 3 d at 25 C. The initial mouth size (z) of the larvae was 0.22–0.23 mm. The newly hatched larvae were 2.02 ± 0.02 mm TL; this increased to 4.12 ± 0.09 mm TL by day 11. By day 54, the larvae had metamorphosed into juveniles and reached 41.12 ± 1.20 mm TL, and by day 93 the juveniles reached 93.78 ± 1.98 mm TL. In all, 49.5% of the larvae were malformed and the type of malformation was diverse.     

Food consumption, utilization, and detoxification enzyme activity of the rice leaffolder larvae after treatment with Dysoxylum triterpenes

The toxicity and physiological (enzyme and nutritional indices) effect of Dysoxylum triterpenes 3β,24,25-trihydroxycycloartane and beddomei lactone were evaluated on the rice leaffolder Cnaphalocrocis medinalis (Guenée). The LC₅₀ [6.66 ppm (SD = 0.31), 5.79 ppm (SD = 0.33) for 3β,24,25-DHCL and BL, respectively] and LC ₉₀ [14.63 ppm (SD = 0.36), 13.49 ppm (SD = 0.27) for 3β,24,25-DHCL and BL, respectively] were identified by probit analysis. Fourth instars were exposed to various concentrations (1.5, 3, 6, and 12 ppm) of Dysoxylum triterpenes. Results showed that treated larvae exhibited reduced food consumption and enzyme activity. Food consumption, digestion, relative consumption rate, efficiency of conversion of ingested food, efficiency of conversion of digested food, and relative growth rate values declined significantly but the approximate digestibility of treated larvae was significantly higher as a result of treatment (in particular 6 and 12 ppm). Likewise, the gut enzymes acid phosphatases, alkaline phosphatases, and adenosine triphosphatases were significantly inhibited by the Dysoxylum triterpenes. The high biological activity of these triterpenes from Dysoxylum sp. could be used as an active principle during the preparation of botanical insecticides for insect pest like rice leaffolder.     

Diversity of swine Bordetella bronchiseptica isolates evaluated by RAPD analysis and PFGE

The degree of genetic diversity in 45 Bordetella (B.) bronchiseptica strains comprised of a vaccine strain (N = 1), reference strains (N = 3) and field isolates (N = 41) was evaluated using random amplified polymorphic DNA (RAPD) fingerprinting and pulsed-field gel electrophoresis (PFGE). Three candidate primers were selected for RAPD analysis after screening 20 random decamer oligonucleotides for their discriminatory abilities. The OPA-07, OPA-08 and OPA-18 primers yielded 10, 10, and 6 distinct fingerprint patterns, respectively. The most common identical RAPD pattern was produced by OPA-07 which was shared by 32 isolates (71.1%), the pattern produced by OPA-08 was shared by 26 isolates (57.8%), and the pattern produced by OPA-18 was shared by 40 isolates (88.9%). The RAPD patterns of the vaccine strain and the 3 reference strains did not match any of the patterns produced by the field isolates when primers OPA-07 and OPA-08 were used. PFGE using the restriction endonuclease XbaI produced a total of 15 patterns consisting of 4 PFGE types (A, B, B1 and C, differing by ≥ 4 bands) and 11 A subtypes (differing by ≤ 3 bands). Most of the field isolates exhibited identical type A and B patterns, suggesting that they were related. The vaccine strain and the three reference strains showed different PFGE patterns as compared to the identical type A strains.