Sequences related to the major subunit gene fedA of F107 fimbriae in porcine Escherichia coli strains that express adhesive fimbriae

Abstract Porcine Escherichia coli strains isolated from cases fo postweaning diarrhea or edema disease were analysed for the presence of fedA , the major subunit gene of F107 fimbriae. The E. coli isolates were known to contain colonisation factor '8813', or to express F107, 2134P or other fimbriae, different from F4, F5, F6, and F41. PCR with fedA -specific primers, restriction enzyme digestion of the PCR product, and nucleotide sequence analysis demonstrated that 2134P pili, colonisation factor '8813' and fimbriae identified on Australian strains of the O141 serotype belong to one family of F107 fimbrial antigens.     

Prevalence of Colonization Factor Antigens (CFAs) and Adherence to HeLa Cells in Enterotoxigenic Escherichia coli Isolated from Feces of Children in São Paulo

Fifty-eight enterotoxigenic Escherichia coli (ETEC) strains, isolated from children with and without diarrhea in Sao Paulo, were examined for the presence of colonization factor antigens (CFAs) and their ability to adhere to HeLa cells. Antisera to CFA/I, the coli surface (CS) antigens CS1CS3, CS2CS3, and CS2 of CFA/II, CFA/III, and CS5CS6 and CS6 of CFA/IV were used. CFAs were identified in 43% of the ETEC strains: 40% of the strains with CFAs harbored CFA/I, 24% carried CFA/II (CS1CS3), 24% carried CFA/IV (CS6), and 12% carried CFA/IV (CS5CS6). CFAs occurred mainly among ETEC strains producing only heat-stable (ST-I) enterotoxin and in strains also producing heat-labile toxin (LT-I). No ETEC strains tested expressed CFA/III. A marked change in serotypes of ST-I-producing strains was found in Sao Paulo between 1979 and 1990. Adherence to HeLa cells was detected in 14% of the ETEC strains. All of them had a diffuse adherence pattern and produced only ST-I, and 88% carried CS6 antigen.     

Cloning and Sequencing of Two New Verotoxin 2 Variant Genes of Escherichia coli Isolated from Cases of Human and Bovine Diarrhea

We cloned and sequenced two new Verotoxin 2 (VT2) variant genes: one from an Escherichia coli strain from a case of bovine diarrhea and the other from an E. coli strain from a patient with diarrhea. The nucleotide and amino acid sequences of these two genes were highly homologous with, but distinct from those of the VT2, VT2vha, VT2vhb, SLT-IIv (VT2vp1) and SLT-IIva (VT2vp2) genes. Their nucleotide sequences were much more closely homologous to that of VT2vh than to that of VT2vp. Search for these two new genes in other Verocytotoxin-producing E. coli strains resulted in the isolation of 2 strains carrying one of the new VT2 variant genes, one strain from Tokyo and the other from Canada.     

Sodium currents in toad cardiac pacemaker cells

Cells in the pacemaker region of toad (Bufo marinus) sinus venosus had spontaneous rhythmic action potentials. The rate of firing of action potentials, the rate of diastolic depolarization and the maximum rate of rise of action potentials were reduced by TTX (10 nm to 1 m). Currents were recorded with the whole cell, tight seal technique from cells enzymatically dissociated from this region. Cells studied were identified as pacemaker cells by their characteristic morphology, spontaneous rhythmic action potential activity that could be blocked by cobalt but not by TTX and lack of inward rectification. When calcium, potassium and nonselective cation currents (I_f) activated by hyperpolarization were blocked, depolarization was seen to generate transient and persistent inward currents. Both were sodium currents: they were abolished by tetrodotoxin (10 to 100 nm), their reversal potential was close to the sodium equilibrium potential and their amplitude and reversal potential were influenced as expected for sodium currents when extracellular sodium ions were replaced with choline ions. The transient sodium current was activated at potentials more positive than –40 mV while the persistent sodium current was obvious at more negative potentials. It was concluded that, in toad pacemaker cells, TTX-sensitive sodium currents contributing both to the upstroke of action potentials and to diastolic depolarization may play an important role in setting heart rate.We thank the Australian National Heart Foundation for their support. D.A.S. is an NHMRC Senior Research Officer.     

Isolation of K88 Antigen Variants (ab,ac, ad) from Porcine Enterotoxigenic Escherichia coli Belonging to Different Serotypes

Fimbriae isolation by means of thermal shock was applied to fifteen K88-positive (three K88ab, nine K88ac and three K88ad) Escherichia coli reference strains belonging to serotypes O8:K87, O32, O45, O138:K81, O141:K85, O147:K89, O149:K91, and O157, as well as to ten K88-positive enterotoxigenic strains isolated from porcine diarrhea in Spain, all of them belonging to the O149 serogroup. Fimbriae were removed from the bacterial cells by thermal shock at 60 C and then precipitated using ammonium sulfate. The final amount of K88 antigen and the purification degree were not related to the serogroup of the bacteria or to the antigen variant but were related to the buffer used for isolation. Phosphate buffer containing urea was shown to be more effective than Tris-HCl for isolation of K88 antigen. The molecular weights by SDS-PAGE for K88ab, K88ac, and K88ad were 28.5, 29.2, and 31.0 kDa, respectively. All enterotoxigenic E. coli strains isolated in Spain showed the K88ac variant.     

痛泻要方对“肝气乘脾”泄泻小鼠肠黏膜及肠内容物消化酶活性的影响

目的探讨痛泻要方对"肝气乘脾"泄泻小鼠肠道酶活性的影响。方法采用"番泻叶-离心管束缚夹尾法"进行"肝气乘脾"泄泻造模,造模成功后以痛泻要方治疗,造模和治疗后分别分析小鼠肠道酶活性。结果造模后,模型组小鼠肠道内容物的淀粉酶活性显著降低(t=4.007,P=0.015),纤维素酶、蛋白酶、蔗糖酶活性下降不显著。模型组小鼠肠黏膜蛋白酶、淀粉酶、蔗糖酶活性显著下降(t_蛋=5.652,P=0.005;Z_淀=-1.964,P=0.050;t_蔗=4.737,P=0.009)。痛泻要方治疗后,中药干预组小鼠肠道内容物蛋白酶、纤维素酶、乳糖酶和蔗糖酶活性变化不显著;自然恢复组的淀粉酶活性显著高于正常组(t=-7.497,P=0.002)。中药干预组小鼠肠道前段黏膜蛋白酶、乳糖酶、淀粉酶、蔗糖酶活性恢复不显著;中药干预组小鼠肠道黏膜中段乳糖酶、蔗糖酶活性显著低于自然恢复组(t_乳=4.074,P=0.013;t_蔗=8.072,P0.001),而蛋白酶、淀粉酶及纤维素酶活性均高于自然恢复组;中药干预组小鼠肠道后段黏膜乳糖酶、蔗糖酶及淀粉酶活性显著高于正常组(t_乳=-7.962,P0.001;t_蔗=-15.921,P0.001;Z_淀=6.489,P=0.034),蛋白酶与纤维素酶活性均有所升高。结论 "肝气乘脾"泄泻肠道内容物及黏膜淀粉酶活性显著降低,痛泻要方对"肝气乘脾"泄泻小鼠肠黏膜乳糖酶、蔗糖酶及淀粉酶活性作用显著。     

Interaction of CP levels in maternal and nursery diets,and its effect on performance,protein digestibility,and serum urea levels in piglets

Reduced protein levels in nursery diets have been associated with a lower risk of postweaning diarrhea, but the interaction with CP levels in maternal diet on the performance of the offspring remains unclear. The objective of this study was to determine the effect of protein content in sow gestation and piglet nursery diets on the performance of the piglets until slaughter. This was studied in a 2 × 2 factorial trial (35 sows, 209 piglets), with higher or lower (H or L) dietary CP in sow diets (168 vs 122 g CP/kg) during late gestation. A standard lactation feed was provided for all sows (160 g CP/kg). For both sow treatments, half of the litters received a higher or lower CP in the piglet nursery diet (210 vs 166 g CP/kg). This resulted in four possible treatment combinations: HH, HL, LH and LL, with sow treatment as first and piglet treatment as second letter. For each phase, all diets were iso-energetic and had a similar level of essential amino acids. P_s*p is the p-value for the interaction effect between sow and piglet treatment. In the nursery phase (3.5–9 weeks of age), a tendency toward interaction between piglet and sow treatments with feed efficiency (P_s*p = 0.08) was observed with HH having the highest gain:feed ratio (G:F) (0.74 ± 0.01), LH the lowest (0.70 ± 0.01) and the other two groups intermediate. In the growing-finishing phase, an interaction was observed between the piglet and sow diets with decreased G:F for LH (P_s*p = 0.04) and a tendency toward interaction with increased daily feed intake for LH (P_s*p = 0.07). The sow diet showed a tendency toward a long-lasting effect on the dressing percentage and meat thickness of the offspring, which was higher for the progeny of H sows (P_s < 0.01 and P_s = 0.02, respectively). At 23 weeks, serum urea concentrations tended to be lower for the HH and LL groups (P_s*p = 0.07). Fecal consistency scores were higher at day 10–day 14 after weaning for piglets from L sows (P_s = 0.03 and P_s < 0.01, respectively). At day 7 after weaning, fecal consistency score was higher for piglets fed the higher protein diet (P_p < 0.01). At 8 weeks of age, the apparent total tract digestibility of CP (ATTD_CP) interacted between piglet and sow diet (P_s*p = 0.02), with HH showing the highest digestibility values. In conclusion, the protein levels in sow late-gestation and piglet nursery diets interacted with feed efficiency, ATTD_CP and serum urea concentrations in the nursery phase.     

葛根芩连汤对泄泻肠道湿热证小鼠肠道双糖酶活性的影响

目的探究葛根芩连汤对泄泻肠道湿热证小鼠肠道双糖酶活性的影响。方法采用“高糖高脂+高温高湿+白酒复合灌胃冰水”的方法制备泄泻肠道湿热证小鼠模型,并运用葛根芩连汤进行干预治疗。分别在干预的第0、2、4、6天无菌采集各组小鼠肠黏膜,运用DNS法测定蔗糖酶活性,ONPG法测定乳糖酶活性,观察葛根芩连汤对泄泻肠道湿热证小鼠肠道双糖酶活性的影响。结果经肠道湿热泄泻造模后,小鼠肠道前段和后段黏膜的乳糖酶和蔗糖酶活性均显著下降,与正常组小鼠相比差异有统计学意义(均P<0.01)。随着治疗时间的延长,治疗组小鼠肠道乳糖酶和蔗糖酶活性与正常组相比有显著的提高(均P<0.05),治疗6 d后,肠道前段黏膜蔗糖酶和乳糖酶活性恢复至正常组水平(P>0.05)。肠道后段乳糖酶活性在第4天时最高,后段黏膜蔗糖酶活性在第6天时最高,与正常组和自愈组相比差异有统计学意义(均P<0.05)。结论泄泻肠道湿热证造模使小鼠肠道黏膜蔗糖酶、乳糖酶等双糖酶的活性显著下降,葛根芩连汤能调节泄泻肠道湿热证小鼠肠道黏膜乳糖酶和蔗糖酶的活性,从而发挥治疗泄泻肠道湿热证的疗效。