Characterization of proteins in sporulated and unsporulated Eimeria maxima oocysts

Proteins in sporulated and unsporulated oocysts of Eimeria maxima were characterized, using monoclonal antibodies (MAB), ELISA, sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), and protein (western) immunoblotting techniques. Three MAB (EM1, EM2, and EM4) were produced against proteins of sporulated oocysts. The ELISA results indicated that EM1 was reactive with sporulated oocyst proteins, EM2 was reactive with sporulated and unsporulated oocyst proteins, and EM4 was reactive with unsporulated oocysts and proteins. Separation of proteins in E maxima sporulated and unsporulated oocysts by SDS-PAGE indicated that sporulated oocysts had proteins of approximately 200 kilodaltons (kD) and distinct protein bands at 21.5 and 45 kD. Using SDS-PAGE, unsporulated oocysts had less-distinct high molecular weight protein bands (greater than 200 kD), compared with sporulated oocysts, and a distinct protein band at 31 kD. Use of all 3 MAB yielded negative results in western blot analysis of fractions obtained by SDS-PAGE.     

日粮中EM添加水平及方式对幼龄獭兔生长性能的影响

试验旨在研究EM添加水平及方式对幼龄獭兔生长性能的影响。将108只50日龄断奶仔獭兔随机分成9组(A-I),每组4个重复,每个重复3只兔子,并置于同笼内饲养。其中,A组为对照组,饲喂基础日粮;B组在基础日粮中添加抗生素;C组则向饮水中添加2‰EM;D、E、F组分别饲喂在基础饲料制粒前添加1‰、2‰、3‰EM的基础日粮;G、H、I组分别饲喂在基础饲料制粒后添加1‰、2‰、3‰EM的基础日粮。试验预饲期5 d,正饲期30 d,每天早晚各饲喂一次。结果表明:制粒前添加2‰EM组(E组)与A、B、C组比较,能极显著提高獭兔生长性能,制粒后添加1‰EM组与制粒前添加2‰EM组差异不显著。建议饲料中EM添加量为:制粒前添加2‰,或制粒后添加1‰。     

Type-I hypersensitivity as a component of eosinophilic myositis (muscular sarcocystosis) in cattle

Eight bovine hearts with lesions of eosinophilic myositis (EM) and 2 bovine hearts without EM lesions were collected at slaughter. Blood samples from these 10 hearts, and the heart of a newborn calf also were collected. Histologically, Sarcocystis cruzi was identified in the 8 hearts with EM lesions and the 2 hearts without EM lesions, but not in the heart of the newborn calf. Serum was harvested from the 10 blood samples and was used in homologous, modified, passive cutaneous anaphylaxis tests. Antigen was prepared from S cruzi bradyzoites isolated from the 2 hearts without EM lesions. Serum samples from the 8 cattle with EM lesions reacted positively to S cruzi antigen. When heat-inactivated IgE in serum (56 C for 4 hours) was used, all passive cutaneous anaphylaxis responses were considered negative. Using ELISA, serum IgE concentrations from the 10 cattle with and without EM lesions were 2.2 to 9 U/ml. As determined by radial immunodiffusion, IgM concentrations were 80 to 215 mg/dl. Immunoglobulin G concentrations were 420 to 2,050 mg/dl, but most were less than or equal to 1,700 mg/dl. Immunoglobulin A concentrations were 0 to 62 mg/dl; 1 steer with EM lesions had 0 mg/dl. Double-gel immunodiffusion confirmed the presence of Sarcocystis-specific precipitating antibodies. Sera from the 10 cattle with and without EM lesions formed at least 1 precipitin band.     

Latex test for rapid rotavirus diagnosis in calves

A latex agglutination test (LA) was compared with direct electron microscopy (EM) for detection of rotavirus infection in calves. A total of 375 samples from 62 calves were collected. Samples were taken when the calves were 1, 3, 5, 7, 10 and 20 days old and some scours samples were collected as well. Altogether 45/375 (12%) specimens were positive in LA and 10/375 (2.7%) were positive in EM. Samples positive in EM were also positive in LA. Out of the 62 calves studied 26/62 (42%) were positive in LA and 8/62 (13%) in EM. We found the LA very easy to perform, to be more sensitive than the EM method and probably a rather specific method for detection of rotavirus infection.     

Antinociceptive effects of epidural buprenorphine or medetomidine, or the combination, in conscious cats

The aim of this study was to compare the antinociceptive effects of epidural buprenorphine (EB), epidural medetomidine (EM) or epidural buprenorphine–medetomidine (EBM). Eight cats were studied. Thermal thresholds (TT) were measured by increasing the temperature of a probe placed on the thorax. Mechanical thresholds (MT) were measured through inflation of a modified blood pressure bladder to the cat's forelimb. After baseline measurements, EB (0.02 mg/kg), EM (0.01 mg/kg) or half of the doses of each drug (EBM) were administered. Data were analysed using anova ( P  < 0.05) and 95% confidence interval (CI). TT increased from 30 min to 1 h after EB and at 45 min after EM. MT increased from 45 min to 2 h after EB, from 15 min to 1 h after EM and at 30, 45 min and at 2 h after EBM. MT were significantly lower after EB than EM at 30 min. TT were above the upper 95%CI from 15 min to 24 h after EB, from 15 min to 4 h after EM and from 15 min to 8 h after EBM. MT were above the upper 95%CI from 15 min to 5 h, and at 8, 12 and 24 h after EB, from 15 min to 6 h after EM and from 15 min to 6 h and at 12 and 24 h after EBM. All treatments had similar onset. Overall, EB presented longer period of action than EBM and EM. The same magnitude of analgesia was achieved, but with fewer side effects when EBM was compared with EM.     

Retrospective study of dystocia in mares at a referral hospital

REASONS FOR PERFORMING STUDY: The period between the onset of dystocia and its resolution has an important bearing on fetal outcome. There are few published data on which to base decisions regarding optimum management of cases in practice. OBJECTIVES: To evaluate and compare the effects of a coordinated dystocia management protocol (CDMP) with that of a previous protocol of random management on time to resolution and outcome in both an emergency dystocia referral population of mares (referred emergency cases: EM) and in a population of mares residing in hospital due to high risk pregnancy (HRP) concerns that then experience dystocia at parturition. METHODS: Retrospective study performed at a university hospital referral centre of cases presenting from 1991-2004 divided into Group 1 (pre-CDMP) and Group 2 (CDMP). RESULTS: Medical records of 71 cases with dystocia were retrieved and data recorded. For referred emergency cases (EM), time from hospital presentation to resolution decreased significantly by 32 min (P = 0.03) after institution of CDMP. Survival rate of mares at discharge was 86%. Survival of EM foals was low, with 10% in Group 1 and 13% in Group 2, surviving to discharge. For EM foals delivered alive, survival to discharge was 30% and 43% in Groups 1 and 2, respectively. Median Stage II was significantly (P < 0.001) different at 71 and 282 min for EM foals delivered alive vs. those not alive at delivery, respectively. Median duration of Stage II was also significantly (P < 0.001) different between EM foals surviving and not surviving to discharge, at 44 and 249 min, respectively. Survival of HRP dystocia foals to discharge was 79%. CONCLUSIONS: Although CDMP reduced the time from presentation at the hospital to resolution significantly for EM, total duration of Stage II for EM was unchanged, as was foal outcome. POTENTIAL RELEVANCE: Very early referral of mares with dystocia to referral centres with dystocia management protocols may improve fetal outcome as increased duration of Stage II in the horse affects fetal outcome negatively.     

Comparison of a commercial enzyme-linked immunosorbent assay with electron microscopy, fluorescent antibody, and virus isolation for the detection of bovine and porcine rotavirus

The purpose in this study was to compare the sensitivity of a commercial enzyme-linked immunosorbent assay (ELISA) with electron microscopy (EM), fluorescent antibody (FA), and virus isolation (VI) for the detection of bovine and porcine rotavirus (RV). Seventy-three bovine and 116 porcine accessions were evaluated by 1 or all 4 diagnostic tests, where suitable specimens were available. For the bovine samples, agreement was 33% between FA and EM, 33% between FA and ELISA, and 92% between EM and ELISA. For the porcine samples, agreement was 79% between EM and FA, 72% between EM and ELISA, and 82% between ELISA and FA. Virus was isolated from 68% and 41% of the bovine and porcine fecal samples, respectively. Commercial ELISA was as sensitive as EM, but was more sensitive than FA or VI for the detection of RV in bovine feces. Electron microscopy was more sensitive than FA, ELISA, or VI for detection of RV in porcine feces. The ELISA was an advantageous alternative to the conventional methods of EM, FA, and VI for the diagnosis of RV in calf feces, but not for porcine feces.     

The role of an early Salmonella Typhimurium infection as a predisposing factor for necrotic enteritis in a laboratory challenge model

Necrotic enteritis (NE) caused by Clostridium perfringens (CP) in poultry is an important bacterial disease in terms of economic implications. The disease is multifactorial and is invariably associated with predisposing factors. In the present experiments, we investigated the potential predisposing role of neonatal Salmonella Typhimurium (ST) infection for NE-associated mortality in a laboratory challenge model. In two experiments, day-of-hatch chicks were randomly assigned to four groups: Group 1, nonchallenged control; Group 2, chickens received Eimeria maxima (EM) and CP; Group 3, chickens received EM and CP and were also challenged with ST at day 1 of age; Group 4, chickens received EM and CP and were also challenged with ST at day 17 of age. Challenged groups received an oral dose of EM at 18 days of age and CP (10(8) colony-forming units/chick) at 22-23 days of age. When compared to EM and CP, chicks challenged with ST (day 1) had increased NE-associated mortality and CP-associated lesion scores (P < 0.05) in both experiments. Furthermore, body weight and body weight gain were lower (P < 0.05) in chicks infected with ST (day 1) in the first experiment, even though no differences (P > 0.05) were observed in weight gain in the second experiment. Chicks challenged with ST (day 17) were similar to the EM and CP group in all of the above-mentioned parameters, indicating that a paratyphoid infection in younger chicks remarkably alters the susceptibility to secondary bacterial infections. Based on this work, the authors suggest that an ST infection early in the age of a chick may be important for altering susceptibility to NE, an observation that may be useful from the perspective of experimental reproduction of this disease and, perhaps, as an economically important reason to address the problem of paratyphoid Salmonella infections in young chicks.     

茶多酚和有效微生物对育成期水貂粪便污染物排放量的影响

为研究饲粮中添加茶多酚(TP)和有效微生物(EM)对育成期水貂粪便污染物排放量的影响,试验选取400只育成前期水貂共160笼,随机分成8组,每组20个重复。采用两因素试验设计,共8个处理组,基础饲粮中分别添加0 EM+0 TP、0 EM+0.05%TP、0 EM+0.1%TP、0 EM+0.2%TP、200 mg/kg EM+0 TP、200 mg/kg EM+0.05%TP、200 mg/kg EM+0.1%TP、200 mg/kg EM+0.2%TP。预试期1周,正试期8周(前4周为试验I期,后4周为试验Ⅱ期)。结果表明:饲粮添加TP不能减少水貂粪便NH3、H2S和CO2生成量;饲粮添加200 mg/kg EM对水貂粪便NH3(48 h)、H2S(72 h和96 h)生成量有显著影响(P0.05);饲粮添加0.1%及0.2%TP可显著降低试验Ⅱ期粪便中氨氮含量(P0.05),饲粮添加200 mg/kg EM可降低粪便中总氮(试验I期)、总磷(试验I和Ⅱ期)、氨氮和亚硝酸盐氮(试验Ⅱ期)含量(P0.05)。由此可见,饲粮添加EM可以减少水貂粪便恶臭气体生成以及粪便污染物的含量,建议水貂饲粮中添加200 mg/kg EM。     

Effects of altered plasma alpha-1-acid glycoprotein levels on pharmacokinetics of some basic antibiotics in pigs: simulation analysis

Effects of altered plasma alpha-1-acid glycoprotein (AGP) levels on pharmacokinetic parameters of basic antimicrobials, erythromycin (EM), lincomycin (LM) and clindamycin (CM) were evaluated in pigs by simulation analysis. Intravenous (i.v.) injections of EM, LM and CM were performed to obtain pharmacokinetic parameters in healthy conditions. Binding parameters were obtained from an in vitro study using ultrafiltration. Simulation studies indicated that an increase of plasma AGP levels resulted in a decrease of both volume of distribution at steady state (Vdss) and total body clearance (Cltot) for all the drugs. Elimination rate constant for LM was almost unchanged by an increase of plasma AGP levels, whereas those for EM and CM were increased. Plasma concentration-time profiles at a high AGP level (often observed in pathophysiological conditions) were also simulated. All of the total plasma concentration-time profiles were different from those at normal AGP level. The differences were characterized by a higher initial concentration with faster or similar elimination. Unbound plasma concentration-time profile of LM was unaffected by AGP levels, whereas EM and CM were eliminated from plasma more rapidly at high AGP level. These results suggested that adjustment of dosage regimen of EM and CM is required in pathophysiological conditions, but that of LM is not required.     

Response of plasma fatty acid profiles to changes in dietary n‐3 fatty acids and its correlation with erythrocyte fatty acid profiles in dogs

An elevated level of long‐chain n‐3 fatty acids (FA) in tissue membranes has a positive influence on the progression and treatment of many diseases. Therefore, dietary supplementation of n‐3 FA is recommended in some diseases. Even though n‐3 FA are absorbed readily from the diet, their incorporation into tissues may be compromised in diseased animals. In a clinical setting, it is desirable to monitor the success of dietary intervention. Plasma FA as well as erythrocyte membrane (EM) FA can be used to monitor dietary FA intake. This study compares FA from EM and plasma with regard to their reaction time and reliability for monitoring dietary changes of tissue FA profiles in dogs. Thirty dogs were divided into three groups and fed for 12 weeks. The control group (CONT) was fed a commercial standard diet low in n‐3 FA. One group received the standard diet and 85 mg/kg body weight of a docosahexaenoic acid (DHA) concentrate (ADD). The third group was fed a commercial dog food containing fish oil (FO), which is rich in eicosapentaenoic acid (EPA). EM and plasma FA profiles were analysed by GC separately. Data on EM FA were published recently. n‐3 FA in plasma reached the new level after 2 weeks (8 weeks in EM). Dietary differences between DHA and EPA are obvious after 1 week already. The concomitant decrease in plasma n‐6 FA differed between ADD and FO. In general, the correlation of n‐6 FA between plasma and EM was low. We therefore conclude that analysis of plasma FA is sufficient for monitoring a diet‐induced increase in tissue n‐3 FA in dogs. However, EM FA should be analysed if the effect of dietary intervention on tissue n‐6 FA is important.     

不同乳酸菌组合对发酵液体饲料品质的影响

本试验用液态发酵法研究不同菌种来源对乳酸菌发酵液体饲料(fermentation liquid feed,FLF) 感官、pH和菌落计数的影响,分别以乳酵素(Lactobacillus enzymes,LE)、乳酵素+基础料(enzymes mixture,EM)、5 mg/kg 乳酸菌种+基础料(spawn mixture,SM) 和空白对照组(control,CON)作为研究对象,所有发酵罐保持发酵底物:水=1:3,(36±1)℃恒温发酵192 h,每隔24 h做1次感官评定和pH测定;取发酵72 h的样品进行培养,分别计数培养48和72 h的菌落。结果表明,EM和SM均具有良好的感官性能,保质期长,其活菌落计数均极显著高于LE与对照组(P<0.01),但EM与SM相比差异不显著(P>0.05);各组的pH均先降低后升高,LE处理组pH显著(P<0.05)或极显著(P<0.01)低于其他试验组。提示,LE发酵方式不能改善FLF品质,EM和SM发酵方式的饲料品质优于LE,EM与SM发酵效果类似。     

Rabbit hemorrhagic disease (RHD)--comparative diagnostic studies using the hemagglutination test and electron microscopy]

A total of 56 liver specimens from rabbits with symptoms of rabbit haemorrhagic disease were tested for virus by electron microscopy (EM) and haemagglutination (HA). Both methods simultaneously gave positive or negative results in 28 or 22 cases, respectively. Divergent results were obtained in only 6 samples. Five of them were positive by EM but negative by HA and in one specimen with a HA-titer of 1:32 virus could not be detected.     

Detection of rotavirus in horses with and without diarrhea by electron microscopy and Rotazyme test

A total of 142 equine fecal samples (93 field fecal and 49 experimental fecal specimens) were examined for rotavirus using direct electron microscopy (EM) and the Rotazyme test. Eighty-six stool specimens were diarrhea samples. The Rotazyme test sensitivity and accuracy as compared to EM was determined by the visual (color reaction) and spectrophotometric methods. The overall agreement was 94.8% and 92.3% between EM and Rotazyme visual and spectrophotometric methods, respectively when suspect reactions (1 + color reaction or net absorbance between 0.05 and 0.1) were not included. The Rotazyme test is a quick, simple, and accurate diagnostic test for detection of rotavirus in equine fecal samples. It could be used by the equine practitioner with a minimum of laboratory facilities and by diagnostic and research laboratories.     

Immobilization of African buffaloes (Syncerus caffer) using etorphine–midazolam compared with etorphine–azaperone

ObjectiveTo compare induction times and physiological effects of etorphine–azaperone with etorphine–midazolam immobilization in African buffaloes.Study designRandomized crossover study.AnimalsA group of 10 adult buffalo bulls (mean body weight 353 kg).MethodsEtorphine–azaperone (treatment EA; 0.015 and 0.15 mg kg^–1, respectively) and etorphine–midazolam (treatment EM; 0.015 and 0.15 mg kg^–1, respectively) were administered once to buffaloes, 1 week apart. Once in sternal recumbency, buffaloes were instrumented and physiological variables recorded at 5 minute intervals, from 5 minutes to 20 minutes. Naltrexone (20 mg mg^–1 etorphine dose) was administered intravenously at 40 minutes. Induction (dart placement to recumbency) and recovery (naltrexone administration to standing) times were recorded. Arterial blood samples were analysed at 5 and 20 minutes. Physiological data were compared between treatments using a general linear mixed model and reported as mean ± standard deviation. Time data were compared using Mann-Whitney U test and reported as median (interquartile range) with p ≤ 0.05.ResultsActual drug doses administered for etorphine, azaperone and midazolam were 0.015 ± 0.001, 0.15 ± 0.01 and 0.16 ± 0.02 mg kg^–1, respectively. Induction time for treatment EA was 3.3 (3.6) minutes and not different from 3.2 (3.2) minutes for treatment EM. The overall mean arterial blood pressure was significantly lower for treatment EA (102 ± 25 mmHg) than that for treatment EM (163 ± 18 mmHg) (p < 0.001). The PaO₂ for treatment EA (37 ± 12 mmHg; 5.0 ± 1.6 kPa) was not different from that for treatment EM (43 ± 8 mmHg; 5.8 ± 1.1 kPa). Recovery time was 0.8 (0.6) minutes for treatment EA and did not differ from 1.1 (0.6) minutes for treatment EM.Conclusions and clinical relevanceTreatment EA was as effective as treatment EM for immobilization in this study. However, systemic arterial hypertension was a concern with treatment EM, and both combinations produced clinically relevant hypoxaemia. Supplemental oxygen administration is recommended with both drug combinations.     

Employing a Monte Carlo algorithm in expectation maximization restricted maximum likelihood estimation of the linear mixed model

Multiple‐trait and random regression models have multiplied the number of equations needed for the estimation of variance components. To avoid inversion or decomposition of a large coefficient matrix, we propose estimation of variance components by Monte Carlo expectation maximization restricted maximum likelihood (MC EM REML) for multiple‐trait linear mixed models. Implementation is based on full‐model sampling for calculating the prediction error variances required for EM REML. Performance of the analytical and the MC EM REML algorithm was compared using a simulated and a field data set. For field data, results from both algorithms corresponded well even with one MC sample within an MC EM REML round. The magnitude of the standard errors of estimated prediction error variances depended on the formula used to calculate them and on the MC sample size within an MC EM REML round. Sampling variation in MC EM REML did not impair the convergence behaviour of the solutions compared with analytical EM REML analysis. A convergence criterion that takes into account the sampling variation was developed to monitor convergence for the MC EM REML algorithm. For the field data set, MC EM REML proved far superior to analytical EM REML both in computing time and in memory need.     

EM微生态制剂对仔猪的保健效果观察

试验开展EM微生态制剂对仔猪增重效果、仔猪腹泻的控制效果、环境除臭及蚊蝇孽生繁殖的消除效果等研究,结果表明:(1)EM制剂对仔猪具有良好的育肥增重效果。试验组平均增重比对照组多1.981 kg,差异显著(P<0.05);(2)EM的添加对仔猪腹泻具有良好的控制效果。试验组死亡率为5%,治愈率为75%;对照组死亡率为40%,自愈率为30%;(3)EM微生态制剂能抑制腐败菌的生长繁殖,有明显消除粪便臭味,减少蚊蝇孽生繁殖,从而净化环境,减少污染,改善仔猪生长环境。结论:EM微生态制剂对仔猪具有保健效果。