迟缓爱德华氏菌检验程序的研究

本试验研究了迟缓德华氏菌（Edwardsiellatrada,Et）的细菌学及致病因子检测,确定了相应检测指标,制定了检验程序。对不同来源的Et作分离及生化鉴定,确定10项生化鉴定指标。用三种方法即平板法（PlateAssay,PA）、接触法（ContactHemolysis,CH）和上清法（SunernatantAs－say,SA）检测Et溶血素,阳性率依次为75％、75％、57．14％。28株Et中,18／26的胞外产物（Extra－cellularProducts,ECP）对HEP－2细胞有细胞毒性,17／28菌株有侵袭力。用ATCC15947株ECP的抗血清进行Det－ELISA,检测阳性率为19／26（73．08％）。     

Essential role of neutrophils but not mammary alveolar macrophages in a murine model of acute Escherichia coli mastitis

Mastitis, the inflammation of the mammary gland, is an important disease affecting dairy animals worldwide. The disease is caused by mammary pathogenic bacteria and Escherichia coli are frequently implicated. Virulence factors of mammary pathogenic E. coli are only partially known and intramammary challenge with LPS elicits neutrophil recruitment in experimental bovine and murine mastitis models. We have previously shown that neutrophil recruitment in LPS-induced murine mastitis is strictly dependent on mammary alveolar macrophages. However, the relative role of alveolar macrophages and blood neutrophils in E. coli mastitis is not well defined. To this end, we selectively depleted mammary alveolar macrophages or blood neutrophils before intramammary challenge with E. coli strain P4 (ECP4). Mice depleted of alveolar macrophages prior to intramammary challenge recruited neutrophils normally and restricted bacterial growth and interstitial invasion. Importantly however, upon depletion of alveolar macrophages, ECP4 invaded the mammary alveolar epithelial cells and formed intracellular bacterial communities. In contrast, neutrophil depletion prior to intramammary infection with ECP4 was associated with unrestricted bacterial growth, tissue damage, severe sepsis and mortality. This study suggests that neutrophils but not alveolar macrophages provide essential antimicrobial defense against mammary pathogenic E. coli. Furthermore, we show here similar invasion after depletion of alveolar macrophages as in our previous studies showing that LPS/TLR4 signaling on alveolar macrophages abrogates ECP4 invasion of the mammary epithelium. Interestingly, similar ECP4 invasion and formation of intracellular communities were also observed following intramammary infection of either iNOS gene-deficient or IL-1 receptor type 1 gene-deficient mice.     

An Escherichia coli phytase expressed in yeast effectively replaces inorganic phosphorus for finishing pigs and laying hens

Two experiments determined the efficacy of an Escherichia coli phytase (ECP) added to P-deficient, corn-soybean meal diets fed to finishing pigs and second-cycle laying hens. Sixty finishing pigs (49 +/- 0.9 kg) were formed into blocks within sex based on weight and ancestry and allotted to a P-deficient diet unsupplemented or supplemented with 0.10% inorganic P (iP) from KH2PO4 or ECP at 250, 500, 1,000, or 10,000 phytase units (FTU)/kg. Individually fed pigs were allowed ad libitum access to the experimental diets until a BW of 120 +/- 3 kg was achieved, at which time the pigs were euthanized and the left fibula and fourth metatarsal were excised for determination of bone ash. Pigs were fed a 2-phase diet program for early- and late-finishing pigs; available P in the basal diets was set 0.10% below the requirement. Dietary supplementation of iP or ECP increased weight gain (P < 0.10) and G:F (P < 0.01); performance was not different (P > 0.13) among the phytase-supplemented groups. Fibula ash was greatest (P < 0.01) for pigs fed diets containing 10,000 FTU of ECP/kg. Two hundred forty second-cycle hens were allotted to a P-deficient diet or a P-deficient diet supplemented with 0.10% iP or ECP at 150, 300, or 10,000 FTU/kg for a 12-wk experiment. The basal diet was a corn-soybean meal diet with no added iP (17% CP, 3.8% Ca, 0.10% available P). Hens fed the P-deficient diet were removed from the experiment after 4 wk due to poor egg production. Supplementation of iP or ECP resulted in increased (P < 0.01) feed intake, egg weight, and egg production during the first 4 wk. During the entire 12-wk period, there were no differences (P > 0.28) between the iP- and ECP-supplemented groups in feed intake, egg weight, or egg production. These experiments reveal that ECP was as efficacious as supplemental iP and that supplementation of an excess dose of ECP was efficacious and without negative effects in finishing pigs and laying hens.     

Resynchronization of estrus in cattle of unknown pregnancy status using estrogen,progesterone, or both

Our objective was to develop treatments applied to cattle of unknown pregnancy status that would resynchronize the repeat estrus of nonpregnant females. In Exp. 1, previously inseminated dairy and beef heifers were assigned randomly to each of three treatments 13 d after AI: 1) no treatment (controls; n = 44); 2) 0.5 mg of estradiol cypionate (ECP) i.m. on d 13 and 20 at the time of insertion and removal of a used intravaginal progesterone (P4)-releasing insert (CIDR; P4 + ECP; n = 44); and 3) same as P4 + ECP without injections of ECP (P4; n = 42). The P4 + ECP (>90%) and P4 (>75%) protocols effectively synchronized repeat periods of estrus to 2 d and did not harm established pregnancies. In Exp. 2, treatments similar to those in Exp. 1 were applied to previously inseminated beef heifers (n = 439). Feeding 0.5 mg of melengestrol acetate (MGA) from d 13 to 19 after AI replaced the CIDR as a source of progestin. Of those heifers not pregnant (n = 65) after the initial AI, more than 86% were reinseminated, but conception was decreased (P < 0.05) by 28 to 39% compared with controls. In Exp. 3, previously inseminated lactating beef cows at four locations were assigned within herd to each of three treatments: 1) no treatment (control; n = 307); 2) same as in Exp. 1, but with P4 + 1 mg of estradiol benzoate on d 13 and 20 (P4 + EB; n = 153); and 3) same as in Exp. 1, P4 + ECP (n = 149). Treatments with P4 plus estrogen did not decrease conception rates in pregnant cows at any location, but increased (P < 0.05) the percentage of nonpregnant cows returning to estrus between 19 and 23 d after timed AI from 29% in controls to 86% in P4 + EB and 65% in P4 + ECP cows. Conception rates at the return estrus were not decreased when treatments occurred between d 13 and 20. In Exp. 4, lactating beef cows were assigned as in Exp. 3 to each of three treatments: 1) no treatment (controls; n = 51); 2) P4 + ECP (n = 47), as in Exp. 1; and 3) a single injection of ECP on d 13 (n = 48). Previously established pregnancies were not harmed (P = 0.70), and return rates of nonpregnant cows did not differ (P = 0.78) among treatments. In summary, in both heifers and lactating beef cows, the P4-based resynchronization treatments increased synchronized return rates when estrus detection rates were low, had no negative effects on established pregnancies, and decreased or tended to decrease conception rates at the resynchronized estrus.     

Effect of estradiol cypionate and GnRH treatment on plasma estradiol‐17β concentrations,synchronization of ovulation and on pregnancy rates in suckled beef cows treated with FTAI‐based protocols

Two experiments were conducted to evaluate the effect of different ovulation inducers on E‐17β plasma concentrations, synchronized ovulations and pregnancy rates. In Experiment 1, cows received a progesterone intravaginal device (PID) with 1 g of progesterone (P4) plus 2 mg of estradiol benzoate (EB) (day 0). At PID removal (day 8), cows received 0.150 mg of D‐cloprostenol and were randomly assigned to four treatment groups (n = 10/treatment): Group ECP: 1 mg of estradiol cypionate at PID removal, Group EB: 1 mg of EB 24 hr after PID removal, Group GnRH: 10 μg of GnRH 48 hr after PID removal, Group ECP‐GnRH: 1 mg of ECP at PID removal plus 10 μg of GnRH 48 hr later. Ultrasonographic examinations were performed to detect the dominant follicle and ovulation. GnRH‐treated cows ovulated later (p < .05) compared to ECP‐ and ECP+GnRH‐treated cows. There were effects of treatment, time and their interaction on E‐17β concentrations (p < .05). ECP treatment affected plasma E‐17β concentration, which increased earlier and decreased later compared to treatments without ECP. In Experiment 2, cows received (i) ECP: n = 126; (ii) EB: n = 126; (iii) GnRH: n = 136; (iv) ECP+GnRH: n = 139; FTAI was performed 48–50 hr after PID removal. Pregnancy rates did not differ among ovulation inducers (p > .05; ECP: 54.0%, 68/126; EB: 49.2%, 62/126; GnRH: 40.4%, 55/136; ECP+GnRH: 43.9%, 61/139). In conclusion, ECP administration (ECP and ECP+GnRH treatments) affected E‐17β concentrations, determining its earlier increase and later decrease compared to treatments without ECP (EB and GnRH treatments). ECP+GnRH‐treated cows achieved the best distribution of ovulations without affecting pregnancy rates.     

Efficacy and equivalency of an Escherichia coli-derived phytase for replacing inorganic phosphorus in the diets of broiler chickens and young pigs

Two studies were conducted to determine the efficacy of an Escherichia coli-derived phytase (ECP) and its equivalency relative to inorganic phosphorus (iP) from monosodium phosphate (MSP). In Exp. 1, one thousand two hundred 1-d-old male broilers were used in a 42-d trial to assess the effect of ECP and iP supplementation on growth performance and nutrient digestibility. Dietary treatments were based on corn-soybean meal basal diets (BD) containing 239 and 221 g of CP, 8.2 and 6.6 g of Ca, and 2.4 and 1.5 g of nonphytate P (nPP) per kg for the starter and grower phases, respectively. Treatments consisted of the BD; the BD + 0.6, 1.2, or 1.8 g of iP from MSP per kg; and the BD + 250, 500, 750, or 1,000 phytase units (FTU) of ECP per kg. Increasing levels of MSP improved gain, gain:feed, and tibia ash (linear, P < 0.01). Increasing levels of ECP improved gain, gain:feed, tibia ash (linear, P < 0.01), apparent ileal digestibility of P, N, Arg, His, Phe, and Trp at d 21 (linear, P < 0.05), and apparent retention of P at d 21 (linear, P < 0.05). Increasing levels of ECP decreased apparent retention of energy (linear, P < 0.01). Five hundred FTU of ECP per kg was determined to be equivalent to the addition of 0.72, 0.78, and 1.19 g of iP from MSP per kg in broiler diets based on gain, feed intake, and bone ash, respectively. In Exp. 2, forty-eight 10-kg pigs were used in a 28-d trial to assess the effect of ECP and iP supplementation on growth performance and nutrient digestibility. Dietary treatments consisted of a positive control containing 6.1 and 3.5 g of Ca and nPP, respectively, per kg; a negative control (NC) containing 4.8 and 1.7 g of Ca and nPP, respectively, per kg; the NC diet plus 0.4, 0.8, or 1.2 g of iP from MSP per kg; and the NC diet plus 500, 750, or 1,000 FTU of ECP per kg. Daily gain improved (linear, P < 0.05) with ECP addition, as did apparent digestibility of Ca and P (linear, P < 0.01). Five hundred FTU of ECP per kg was determined to be equivalent to the addition of 0.49 and 1.00 g of iP from MSP per kg in starter pigs diets, based on ADG and bone ash, respectively.     

Stimulation of Ovulation in Seasonally Anovulatory and Vernal Transitional Mares with Estradiol and Domperidone: Dose and Combination Studies

Three experiments were conducted to test the efficacy of different doses of estradiol cypionate (ECP) and domperidone for inducing ovulation in seasonally anovulatory (January; Experiments 1 and 2) and transitional period (March; Experiment 3) mares. In the first two experiments, mares in Kentucky and Louisiana were administered domperidone (3 g in biodegradable particles) alone or after pretreatment with 100 or 150 mg of ECP; another group received ECP, domperidone, and progesterone, and a fifth group received ECP and progesterone only (the latter two in Kentucky). Control mares in both states received no treatment. The proportion of mares ovulating within 35 days (for mares treated in January) was greater (P = .0002) for those receiving any combination of ECP plus domperidone relative to mares not receiving the combined treatment. Addition of progesterone to ECP plus domperidone did not enhance (P = .7) the response relative to the combination alone; domperidone by itself, or ECP plus progesterone, did not alter the response relative to controls (P > .24). Experiment 3 was conducted in Louisiana as a 2 × 3 factorial, with two doses of domperidone (1.5 or 3 g) and three doses of ECP (0, 75, or 150 mg). There was no main effect of domperidone or ECP dose; a greater proportion (P = .055) of mares receiving any combination of ECP plus domperidone ovulated in 21 days compared with those receiving no ECP. In conclusion, pretreatment with ECP before injection of domperidone 10 days later can be used to increase the proportion of mares ovulating early in the year; within the limits of the present experiments, there appears to be no difference in doses of ECP of 75, 100, or 150 mg, and no difference in doses of domperidone of 1.5 or 3 g.     

The humoral immune response of rainbow trout (Salmo gairdneri) immunised by various regimes and preparations of Aeromonas salmonicida antigens

Antibody production in rainbow trout to extracellular antigens was investigated. The following antigen preparations and immunisation regimes were used: native extracellular products (ECP) in Freund's complete adjuvant (FCA), intraperitoneally (i.p.) with and without booster; formalinized ECP in FCA, i.p. with and without booster; washed, formalinized A. salmonicida cells in FCA, i.p., with booster; native ECP in saline, i.m., four weekly injections at two different doses, 45 micrograms and 6 micrograms each injection (after the protocol of Shieh, 1985). Using crossed normal rainbow trout serum, i.p., single injection (after the protocol of Sakai, 1985). Using crossed immunoelectrophoresis all antisera contained precipitating antibodies to three to five ECP components except that from fish immunised i.m. with 6 micrograms protein where antibodies were undetectable. In no case were specific antibodies to ECP protease or haemolysin detected. In a rabbit immunised with formalinized ECP in FCA under a similar regime to the rainbow trout, antibodies to at least 15 ECP components, including protease and haemolysin, were detected. The assumption of a specific immune response to the protease, at least in respect of antibody production, in recent reports of protection afforded by vaccines composed of either crude ECP or partially purified protease (Shieh, 1985) or partially purified protease inactivated by normal serum (Sakai, 1985) is not supported by the present findings.     

The effect of estradiol cypionate (ECP) on ovarian follicular development and ovulation in dairy cattle.

The efficacy of estradiol cypionate (ECP) for synchronizing ovarian follicular development was determined in lactating Holstein-Friesian cattle. In Experiment 1, 13 cattle were given simultaneous intramuscular (i.m.) injections of 100 mg progesterone and 0 (control), 0.5 or 1.0 mg ECP on Day 3, after a synchronized ovulation (Day 0). Maximum diameter of the dominant follicle of Wave 1 was significantly larger in control cattle than in those given 0.5 or 1.0 mg ECP (means: 15.7, 13.2, and 12.9 mm, respectively). Mean day of emergence of Wave 2 was significantly later in controls than in those given 1.0 mg ECP, with the 0.5 mg group intermediate (Days 10.2, 8.8 and 9.5, respectively). In Experiment 2, 14 cattle were given a CIDR-B and IM injections of 1 mg ECP and 50 mg progesterone without regard to stage of cycle (treatment = Day 0). On Day 8, the CIDR-B was removed and 500 micrograms cloprostenol injected, IM. Mean days of wave emergence (Day 3.4; range: -2 to 7) and ovulation (Day 12.1; range: 10 to 14) indicated that ECP had limited efficacy for synchronizing follicular development and ovulation in dairy cattle when given at random stages of the estrous cycle.     

Effects of oestradiol cypionate on spontaneous and oxytocin-stimulated postpartum myometrial activity in the cow

The effects of oestradiol cypionate (ECP) on spontaneous and oxytocin-induced postpartum myometrial activity were measured in four cows using strain gauge transducers (SGT). On the first day after parturition, prior to treatment with ECP, myometrial activity consisted mainly of single-peak contractions (mean frequency 9.6/h, mean duration 141.0s, the majority of the contractions being propagated in a tubocervical direction. Injection of ECP (5 mg i.m.) 18 h after parturition led to suppression of coordinated myometrial activity and the development of sustained low amplitude contractions of reduced frequency (mean 2.9/h, P less than 0.01) and increased duration (mean 422.2 s, P less than 0.05), with multiple superimposed small peaks. In addition, all parts of the uterus tended to contract simultaneously. These changes were apparent by 4 h after treatment and persisted until day 5 after parturition. Injection of oxytocin (25 USP units i.v.) at 24 h after parturition stimulated the reappearance of single-peak coordinated contractions. However, pretreatment with ECP did not enhance the myometrial response to oxytocin.     

Effect of Drinking-Water Administration of Experimental Chlorate Ion Preparations on Salmonella enterica serovar Typhimurium Colonization in Weaned and Finished Pigs

Foodborne disease caused by Salmonella is of public health and economic significance. In order to assess the practical effectiveness of a new intervention strategy, experimental chlorate preparations (ECP) were administered via the drinking water to weaned and finished pigs that had been orally challenged the previous day with 10(9)-10(10) colony-forming units of Salmonella serovar Typhimurium. After 24 or 36 h ad libitum access to 0X, 1X or 2X ECP treatment (where X is the concentration estimated to deliver a minimal daily effective dose), the pigs were euthanized and gut contents and lymph tissue collected at necropsy were cultured for the challenge Salmonella. Drinking water administration of ECP effectively reduced (p < 0.05) caecal Salmonella concentrations and, with the weaned pigs, tended (p < or = 0.10) to reduce rectal Salmonella concentrations. No negative effects of ECP treatment on water intake and animal wellbeing were observed and only marginal effects on gut fermentation characteristics occurred. The bactericidal effect of administering ECP in drinking water was relatively rapid, with reductions in caecal Salmonella concentrations occurring within 24 h. These results suggest that ECP administered to pigs just days before slaughter may reduce gut concentrations of Salmonella; however, the impacts of such reductions on slaughter hygiene have yet to be determined.     

围栏封育下华北半干旱草原植物生态位研究

为查明围栏封育条件下华北半干旱草原植物生态位格局及变化,于河北坝上地区红松洼国家级自然保护区设置100 m×100 m围栏监测固定大样地,采用样方法对封育区和放牧区植被生长状况进行调查,运用生态位理论对围栏内外主要植物的生态位宽度、生态位重叠值和生态位相似比例进行测算.结果表明:披针叶苔草(Carex lanceolata)在围栏内外的生态位宽度均为最大.披针叶苔草、裂叶蒿(Artemisia tanacetifolia)、地榆(Sanguisorba officinalis)、毛茛(Ranunculus japonicus)、珠芽蓼(Polygonum viviparum)等生态位宽度较大物种之间具有较高程度的生态位重叠.这些物种的生态位相似比例也较高;而生态位宽度较小的物种其生态位相似比例较小.表明生态位宽度与生态位重叠值、生态位宽度与生态位相似比例之间存在一定的线性关系.     

Efficacy of an E. coli phytase expressed in yeast for releasing phytate-bound phosphorus in young chicks and pigs

Four chick trials and one pig trial were conducted to investigate the phosphorus-releasing efficacy oftwo commercial phytase enzymes (Natuphos and Ronozyme) and an experimental E. coli phytase enzyme (ECP) when added to corn-soybean meal diets containing no supplemental inorganic P (iP). In the 13- or 14-d chick trials, three or four graded levels of iP (0, 0.05,0.10,0.15%) from KH2PO4 were added to the basal diet to construct standard curves from which bioavailable P release could be calculated for the phytase treatments. In all cases, phytase supplementation levels were based on an assessment of phytase premix activity (i.e., P release from Na phytate at pH 5.5). Linear (P < 0.01) responses in tibia ash and weight gain resulted from iP supplementation in all assays. In the first chick trial, supplementation of 500 phytase units (FTU)/kg of ECP resulted in superior (P < 0.01) weight gain and tibia ash values compared with 500 FTU/kg of Natuphos. Results of the second chick trial revealed P-release values of 0.032 and 0.028% for 500 FTU/kg Natuphos and Ronozyme, respectively, and these were lower (P < 0.01) than the 0.125% P-release value for 500 FTU/kg of ECP. Tibia ash responded quadratically (P < 0.05) in response to graded levels of ECP up to 1,500 FTU/kg in the third chick trial. Combining Natuphos with either Ronozyme or ECP in Chick Trial 4 revealed no synergism between phytases with different initiation sites of P removal. The pig trial involved 10 individually fed weanling pigs per diet, and and phytase enzymes were supplemented to provide 400 FTU/kg in diets containing 0.60% Ca. Based on the linear regression of fibula ash on supplemental iP intake (r2 = 0.87), P-release values were 0.081% for Natuphos, 0.043% for Ronozyme, and 0.108% for ECP. These trials revealed an advantage of the E. coli phytase over the commercial phytases in young chicks.     

紫花苜蓿根际土壤细菌群落对腐熟牛粪响应

为了探讨腐熟牛粪施用量对作物根际土壤细菌丰度、多样性及群落结构的影响,本研究以盆栽紫花苜蓿(Medicago sativa L.)为试验材料,分别添加0％,3％,6％,9％的腐熟牛粪,在添加30 d,60 d,90 d后进行取样,利用Illumina-Miseq高通量测序平台对土壤细菌16S rRNA V3-V4区进行...     

Oestradiol-17β plasma concentrations after intramuscular injection of oestradiol benzoate or oestradiol cypionate in llamas (Lama glama)

Background

Llamas (Lama glama) are induced ovulators and the process of ovulation depends on dominant follicular size. In addition, a close relationship between behavioural estrus and ovulation is not registered in llamas. Therefore, the exogenous control of follicular development with hormones aims to predict the optimal time to mate. Oestradiol-17β (E₂) and its esters are currently used in domestic species, including camelids, in synchronization treatments. But, in llamas, there is no reports regarding the appropriate dosages to be used and most protocols have been designed by extrapolation from those recommended for other ruminants. The aim of the present study was to characterize plasma E₂ concentrations in intact female llamas following a single intramuscular (i.m.) injection of two oestradiol esters: oestradiol benzoate (EB) and oestradiol cypionate (ECP).

Methods

Twelve non pregnant and non lactating sexually mature llamas were i.m. injected on day 0 with 2.5 mg of EB (EB group, n = 6) or ECP (ECP group, n = 6). Blood samples were collected immediately before injection, at 1, 6, 12, 24 h after treatment and then daily until day 14 post injection. Changes in hormone concentrations with time were analyzed in each group by analysis of variance (ANOVA) using a repeated measures (within-SS) design. Plasma E₂ concentrations and area under the concentration-time curve (AUC) values were compared between groups by ANOVA. In all cases a Least-Significant Difference test (LSD) was used to determine differences between means. Hormonal and AUC data are expressed as mean ± S.E.M.

Results

Peak plasma E₂ concentrations were achieved earlier and were higher in EB group than in ECP group. Thereafter, E₂ returned to physiological concentrations earlier in EB group (day 5) than in ECP group (day 9). Although plasma E₂ profiles differed over time among groups there were no differences between them on AUC values.

Conclusions

The i.m. injection of a single dose of both oestradiol esters resulted in plasma E₂ concentrations exceeding physiological values for a variable period. Moreover, the plasma E₂ profiles observed depended on the derivative of oestradiol administered. This basic information becomes relevant at defining treatment protocols including oestrogens in llamas.     

Comparison of various amino acids for estimation of microbial nitrogen in digesta

Methods of estimating microbial N in digesta were compared, and two simplified methods were described for separation of diaminopimelic acid (DAP) from methionine in acid hydrolysates of biological samples using an automated amino acid analyzer. Recoveries of DAP when added to bacterial, abomasal and feed samples were 92 to 98%. All samples analyzed (bacteria, protozoa and several feedstuffs) contained detectable DAP (or another amino acid with similar elution time). Apparent DAP-N (percentage of total sample N) in feeds ranged from 12 to 78% of bacterial DAP-N concentration. These values are greater than can be accounted for by bacterial contamination. Correction for apparent dietary DAP was appropriate when using this microbial marker. Reasonable estimation of bacterial N was made in abomasal digesta of steers fed high-grain diets using DAP, lysine or combinations of DAP and leucine or lysine and leucine. There was excellent agreement among these marker methods in ranking dietary treatment regimens with respect to bacterial N present in the abomasum. The use of leucine alone, or the combination of DAP and lysine, gave nonrealistic values. Two-amino-ethylphosphonic acid (AEP) was not detected in acid hydrolysates of mixed ruminal protozoa or abomasal digesta. Two other ninhydrin-reactive compounds with elution times close to AEP were detected in all samples analyzed, which may have been identified as AEP in other studies. It was concluded that corrected DAP and lysine were the best estimates of bacterial N and AEP was precluded as a marker of protozoal N.     

迟缓爱德华氏菌胞外产物的细胞毒性和动物致病性

分析了２８株迟缓受德华氏菌（Ｅdwardwiella tarda,Ｅt）胞外产物（extracellular puodrcts,ＥＣＰ）的致病性。将Ｅt用覆有玻璃纸的ＴＳＡ培养,１８株（６９．２３％）的 ＥＣＰ使ＨＥp－２细胞产生病认,认圆,脱落。这种细胞毒作用能被ＡＴＣＣ１５９４７株的ＥＣＰ抗体作为探 针进行免疫转印分析,结果显示,不同来源的９株Ｅt的ＥＣＰ在 转印膜上出现３条共同蛋白条带,相对分子质量分别约１４４００、３２、３００、７９、     

Semen parameters and hormone concentrations in stallions subjected to long-term estrogen administration

Eight pony stallions were paired by estimated daily sperm output (DSO) and randomly assigned to one of two treatments in a randomized block experiment. Stallions received 44 μg/kg BW estradiol cypionate (ECP) or an equivalent volume of physiological saline solution on alternate days during the breeding season. Blood samples collected immediately preceding each injection were assayed for luteinizing hormone (LH), estradiol-17β (E₂) and testosterone (T). Semen was collected twice weekly, 3.5 days apart, to evaluate sperm motility and total number of sperm per ejaculate. Prior to and after 4, 8 and 12 weeks on treatment, semen was collected once daily for 7 days to determine DSO. Data were separated into 9 periods (10 days each) for statistical analysis and subjected to analysis of variance for a randomized block design to determine treatment effects.There were no differences (p>.05) between groups for DSO or LH prior to initiation of treatment. Testosterone was higher (p<.05) in ECP stallions compared with C stallions prior to treatment and at all time points measured. As expected, E₂ was higher (p<.05) in the ECP stallions compared to C stallions after 20 days (period 2) of treatment and for the remainder of the experiment. However, E₂ was higher (p<.05) in the C group prior to treatment, but there was no difference between the groups after 10 d of treatment (period 1). ECP stallions had higher (p<.05) DSO than C stallions after 30 d on treatment. After 40 and 50 d (periods 4 and 5), ECP stallions demonstrated higher (p<.05) total sperm per period than C stallions. This was preceded by higher (p<.05) LH values for ECP stallions than for C stallions after 10 and 20 d (periods 1 and 2). No differences were found between the ECP and C groups for LH between 30 and 60 d. Although numerically higher, no significant differences (p>.05) were seen after 60 days for DSO or after 60, 70 or 80 days for total sperm per period. ECP stallions had higher (p<.05) DSO and total sperm per period after 90 d than C stallions. Additionally, LH remained significantly higher (p<.05) in the ECP group after 60 days (periods 7, 8 and 9). Elevated LH concentrations in ECP stallions demonstrated that estrogen treatment did not inhibit LH secretion in this study.     

蚯蚓体腔液及粗组分体外抗菌特性

利用钢圈法测定蚯蚓体腔液对大肠杆菌(Escherichia coli)、绿脓杆菌(pseudodomonas aeru ginosa )和枯草芽孢杆菌(Bacillus subtilis)的抑制作用，并利用微量平板稀释法测定蚯蚓体腔液对绿浓杆菌的最低抑菌浓度。结果如下：蚯蚓体腔液对枯草芽孢杆菌和大肠杆菌没有抑制作用、对绿脓杆菌有抑制作用，浓度为0．1g／ml时绿脓杆菌的抑菌圈直径为1．53cm；蚯蚓体腔液对绿脓杆菌的最低抑菌浓度(MIC)为6．25mg／mL。体腔液用85％的硫酸铵沉淀、截留分子量为10kDa和1kDa的超滤膜进行超滤，得到分子量分别为大于10kDa、1kDa～10kDa、小于1kDa的三种组分ECP1、ECP2、ECP3。用钢圈法测定了ECPl、ECP2、ECP3对绿脓杆菌的抑制作用，其中ECP3的抑菌作用最强，ECP1、ECP2和ECP3的抑菌圈直径分别为7．68mm、11．44mm和16．8mm。ECP3对绿脓杆菌的最低抑菌浓度为0．018mg／mL。     

Antigenic variability in bovine viral diarrhea virus (BVDV) isolates from alpaca (Vicugna pacos), llama (Lama glama) and bovines in Chile

Llamas and alpacas are domesticated South American camelids (SACs) important to ancestral population in the Altiplano region, and to different communities where they have been introduced worldwide. These ungulates have shown to be susceptible to several livestock viral pathogens such as members of the Pestivirus genus and mainly to bovine viral diarrhea virus (BVDV). Seventeen Chilean BVDV isolates were analyzed by serum cross neutralization with samples obtained from five llama, six alpacas, three bovines, plus three reference strains belonging to different subgroups and genotypes. The objective was to describe antigenic differences and similarities among them. Antigenic comparison showed significant differences between different subgroups. Consequently, antigenic similarities were observed among isolates belonging to the same subgroup and also between isolates from different animal species belonging the same subgroup. Among the analyzed samples, one pair of 1b subgroup isolates showed significant antigenic differences. On the other hand, one pair of isolates from different subgroups (1b and 1j) shared antigenic similarities indicating antigenic relatedness. This study shows for the first time the presence of antigenic differences within BVDV 1b subgroup and antigenic similarities within 1j subgroup isolates, demonstrating that genetic differences within BVDV subgroups do not necessary corresponds to differences on antigenicity.