PCR对猪全血、血清、尿液和粪便中PCV2的检测

根据已发表的基因序列 ,选取 PCV2保守的基因片段 ,利用计算机软件设计的一对引物对某猪场的 7只患病仔猪的全血、血清、尿液、和粪便进行 PCR扩增 ,再对扩增产物酶切鉴定 ,并将 PCR产物连接到 p MD1 8-T载体上 ,然后转化到大肠杆菌 DH5α感受态细胞中 ,抽提重组质粒 ,对重组质粒 PCVp MD1 8进行 PCR、酶切鉴定及测序。结果表明 :全血、血清、尿、粪便阳性率分别为 85.7% (6/7)、71 .4% (5/7)、42 .9%(3 /7)、2 8.6% (2 /7)。通过比较 ,全血的阳性率最高 ,其次为血清     

Total observed error,total allowable error,and QC rules for canine serum and urine cortisol achievable with the Immulite 2000 Xpi cortisol immunoassay

Determining a simple quality control (QC) rule for daily performance monitoring depends on the desired total allowable error (TEa) for the measurand. When no consensus TEa exists, the classical approach of QC rule validation cannot be used. Using the results of previous canine serum and urine cortisol validation studies on the Immulite 2000 Xpi, we applied a reverse engineering approach to QC rule determination, arbitrarily imposing sigma = 5, and determining the resulting TEa for the QC material (QCM; TEa_QCM) and the resulting probability of error detection (P_ed) for each QC rule. For the simple QC rule 1_2.5S with P_ed = 0.96 and probability of false rejection (P_fr) = 0.03, the associated TEa_QCM were 20% and 35% for serum and 28% and 24% for urine QCM1 and QCM2. If these levels of TEa_QCM are acceptable for interpretation of patient sample results, then users can internally validate the 1_2.5S QC rule, provided that their QCM CVs and biases are similar to ours. Otherwise, more stringent QC rules can be validated by using a lower sigma to lower the TEa_QCM. With spiked samples (relevant cortisol concentrations in the veterinary patient matrix) at 38.6 and 552 nmol/L of cortisol, TEa_QCM at sigma = 5 were much higher (54% and 40% for serum; 90.3% and 42.8% for urine). Spiked samples generate TEa that is probably too high to be suitable for daily QC monitoring; however, it is crucial to verify spiked sample observed total error (TEo; 26% and 18% for serum, 60% and 30% for urine) < TEa_QCM, and to use spiked sample TEo for patient result interpretation. In the absence of consensus TEa for cortisol in dogs, we suggest the use of a 1_2.5S rule, provided that users accept the associated level of TEa_QCM also as clinical TEa for results interpretation.     

Evaluation of urine and serum metabolites in miniature schnauzers with calcium oxalate urolithiasis.

To evaluate underlying causes of calcium oxalate urolithiasis, 24-hour excretion of urine metabolites was measured in 6 Miniature Schnauzers that formed calcium oxalate (CaOx) uroliths during periods when they were fed a standard diet and during periods when food was withheld. Serum concentrations of parathyroid hormone and 1,25-dihydroxyvitamin D also were evaluated. Serum calcium concentrations were normal in all 6 affected Miniature Schnauzers; however, during diet consumption, mean 24-hour urinary excretion of calcium was significantly (P = 0.025) higher than calcium excretion when food was withheld. In 1 dog, urinary calcium excretion was lower during the period of food consumption, compared with the period when food was withheld. Compared with clinically normal Beagles, Miniature Schnauzers that formed CaOx uroliths excreted significantly greater quantities of calcium when food was consumed (P = 0.0004) and when food was withheld (P = 0.001). Miniature Schnauzers that formed CaOx uroliths excreted significantly less oxalate than clinically normal Beagles during fed (P = 0.028) and nonfed (P = 0.004) conditions. Affected Miniature Schnauzers also excreted abnormally high quantities of uric acid. Excretion of citrate was not different between Miniature Schnauzers with CaOx urolithiasis and clinically normal Beagles. In 5 of 6 Miniature Schnauzers with CaOx urolithiasis, concentrations of serum parathyroid hormone were similar to values from age- and gender-matched Miniature Schnauzers without uroliths. The concentration of serum parathyroid hormone in 1 dog was greater than 4 times the mean concentration of clinically normal Miniature Schnauzers. Mean serum concentrations of 1,25-dihydroxyvitamin D in Miniature Schnauzers with calcium oxalate urolithiasis were similar to concentrations of clinically normal Miniature Schnauzers.     

Comparison of cortisol concentrations in saliva and plasma of dogs

It is increasingly important to have simple, non-invasive indicators of stress in animals. Studies in various species have shown that concentrations of cortisol in saliva relate closely to plasma levels of the free hormone; the aim of the present procedure was to show a similar correlation in the dog. Baseline blood and saliva samples were collected concurrently from six male beagles. Synthetic adrenocorticotrophic hormone was given and further samples were collected at 0.25, 0.5, one, two and 2.5 hours later. The results indicated a statistically significant correlation between the levels of cortisol in blood and saliva. Concentrations in saliva were between 5 and 10 per cent of those in plasma at each collection time. To demonstrate a response to a more natural stimulus, saliva samples were taken from a dog during exposure to a known stressor for that individual. The results showed a marked, delayed increase from baseline which was maintained for at least 0.5 hours after stressing.     

Variations of serum cortisol in Argentine horses subjected to ship transport and adaptation stress

The response to stress after transport by ship and adaptation to a new environment was evaluated by variations of serum cortisol (RIA method) in 40 horses imported in Italy from Argentina. The study started at arrival (April 2000) and lasted 6 months (October 2000). As a control group, 42 horses living in Italy have been evaluated. In all subjects, hematocrit, total protein, total bilirubin, urea, creatinine, and creatinine kinase were determined. In control horses, cortisol values ranged between 103 and 278 nM/L (mean 190.24 ± SE 13.50, SD 43.82, CV 23.03), and no significant differences among sex or age groups were observed (P > .05). In subjects coming from Argentina, cortisol values observed at landing were within the normal range (193.73 nM/L ± SE 7.95, SD 50.29, CV 25.95), increased substantially in May (366.06 nM/L ± SE 10.25, SD 64.81, CV 17.70) and in June (337.19 nM/L ± SE 12.16, SD 76.89, CV 22.80) and returned to normal in October (187.87 nM/L ± SE 12.54, SD 43.50, CV 23.15). The differences observed in May and in June were significant in comparison with the values observed in April (P < .01). The variations of CK in horses coming from Argentina were of great interest because they seemed at high levels in May and June. Data indicated that adaptation to the journey was very good. Stress conditions appeared in the first period of stay in Italy, maybe because of hierarchical conflicts resulting from an incorrect grouping of the subjects.     

Evaluation of serum cortisol, metabolic parameters, acute phase proteins and faecal corticosterone as indicators of stress in cows

To assess the validity of laboratory parameters in blood and faeces as indicators of stress in cows, concentrations of cortisol, non-esterified fatty acids (NEFAs), 3-hydroxybutyrate, glucose, triglycerides, cholesterol, serum amyloid A (SAA) and haptoglobin in serum, as well as corticosterone in faeces, were determined in two breeds of cattle (Alberes and Bruna dels Pirineus) under different systems of housing and feeding. Serum cortisol concentrations were markedly elevated in the Alberes group, probably because they were less habituated to human handling. Corticosterone concentrations in faeces were significantly increased in the Bruna dels Pirineus cattle on Alberes pastures. Concentrations of NEFAs and cholesterol were significantly elevated in the Alberes cows, indicating an adrenergic stimulus of lipolysis or the existence of nutritional stress. SAA concentrations were significantly higher in groups living in hardy conditions, whereas there were no significant differences in haptoglobin between the three groups.     

强冷应激对阿勒泰羔羊血清IgG、皮质醇及小肠免疫相关细胞的影响

为了探讨较强寒冷刺激对阿勒泰羔羊血清IgG和皮质醇含量的影响及小肠黏膜免疫屏障的适应特征。把阿勒泰羔羊置于-30～-35℃分别进行0,6 h、1,4,12,24 d的冷刺激,利用酶联免疫吸附试验(ELISA)对阿勒泰羔羊血清中IgG和皮质醇的含量进行测定;通过石蜡切片和HE染色对阿勒泰羔羊小肠的显微结构进行观察;并通过改良甲苯胺蓝染色鉴定小肠中的甲苯胺蓝染色阳性细胞。发现冷应激后IgG质量浓度都比正常组明显降低,其中冷应激4 d与正常组质量浓度差异极显著(P0.01),冷应激后皮质醇质量浓度比正常组表达上升,其中冷应激6 h时与正常组差异显著(P0.05),而24 d与正常组差异极显著(P0.01);IgG与皮质醇浓度在4,12和24 d呈较强正相关,6 h和1 d呈负相关,说明在强冷应激时,绵羊机体免疫机能的调节及抗炎症反应等受到了抑制。冷应激后十二指肠、空肠和回肠的显微结构发生了相应变化,其中最明显的是肠绒毛变短且变粗,回肠内淋巴小结数量增多;对小肠免疫相关细胞而言,同一温度下各肠段中细胞数量呈趋势变化,上皮内淋巴细胞和杯状细胞的数量在十二指肠、空肠和回肠中呈上升趋势,而肥大细胞数量呈下降趋势;冷应激后淋巴细胞数量较常温有所减少,杯状细胞和肥大细胞数量较常温下都有所增加,说明在强冷应激下机体通过激素的释放努力适应环境变化的同时,也会使小肠免疫功能下降。     

Detecting and quantifying marijuana metabolites in serum and urine of 19 dogs affected by marijuana toxicity

Veterinarians diagnose marijuana toxicity based on clinical signs and history, or in conjunction with an over-the-counter (OTC) human urine drug screen. With the legalization of recreational marijuana use becoming more prevalent in the United States, a more accurate test to aid in the diagnosis of canine marijuana toxicity is needed. We collected urine and serum samples from 19 dogs with confirmed or suspected marijuana toxicosis from multiple veterinary hospitals and analyzed them with a novel UPLC-MS/MS method. Calibrations from 0.1 to 100 ng/mL and QC materials were prepared. Samples were extracted, purified, and eluted with solid-phase extraction. Urine samples were tested with an OTC human urine drug screen. The limit of detection (LOD) and lower limit of quantification (LLOQ) ranges for marijuana metabolites in serum were 0.05–0.25 ng/mL and 0.1–0.5 ng/mL, respectively. In urine, the LOD and LLOQ ranges for the metabolites were 0.05–0.1 ng/mL and 0.1–0.5 ng/mL, respectively. In serum, median and range of metabolite concentrations (ng/mL) detected included: THC, 65.0 (0.14–160); 11-OH-Δ⁹-THC, 4.78 (1.15–17.8); 11-nor-9-carboxy-Δ⁹-THC, 2.18 (0.71–7.79); CBD, 0.28 (0.11–82.5); and THC-glucuronide, 2.05 (0.72–18.3). In the 19 urine samples, metabolite: creatinine (ng: mg) values detected included: THC, 0.22 (0.05–0.74); 11-OH-Δ⁹-THC, 0; 11-nor-9-carboxy-Δ⁹-THC, 1.32 (0.16–11.2); CBD, 0.19 (0.12–0.26); THC-COOH-glucuronide, 0.08 (0.04–0.11); and THC-glucuronide, 0.98 (0.25–10.7). Twenty of 21 urine samples tested negative for THC on the urine drug screen. All 19 serum samples contained quantifiable concentrations of THC using our novel UPLC-MS/MS method. Utilizing a UPLC-MS/MS method can be a useful aid in the diagnosis of marijuana toxicosis in dogs, whereas using an OTC human urine drug test is not a useful test for confirming marijuana exposure in dogs because of the low concentration of THC-COOH in urine.     

Effects of purified zearalenone on growth performance, organ size, serum metabolites, and oxidative stress in postweaning gilts

Zearalenone (ZEA), an estrogenic mycotoxin, is produced mainly by Fusarium fungi. Previous studies indicated that acute ZEA exposure induced oxidative stress and damage in multiple organs. Therefore, the present study was designed to investigate the adverse effects of dietary ZEA (1.1 to 3.2 mg/kg of diet) on oxidative stress and organ damage in postweaning gilts. A total of 20 gilts (Landrace × Yorkshire × Duroc) weaned at d 21 with an average BW of 10.36 ± 1.21 kg was used in the study. Gilts were housed in a temperature-controlled room, divided into 4 treatments, and fed a basal diet only (control) or basal diet supplemented with purified ZEA at a dietary concentration of 1 (ZEA1), 2 (ZEA2), or 3 (ZEA3) mg/kg of diet for 18 d ad libitum. The actual ZEA contents (analyzed) were 0, 1.1 ± 0.02, 2.0 ± 0.01, and 3.2 ± 0.02 mg/kg for control, ZEA1, ZEA2, and ZEA3, respectively. Gilts fed different amounts of dietary ZEA grew similarly with no difference (P > 0.05) in feed intake. Vulva size increased linearly over the 18 d of feeding in gilts fed diets containing 1.1 mg of ZEA/kg or greater (P < 0.001). Relative weight of genital organs, liver, and kidney increased linearly (P < 0.05) in a ZEA-dose-dependent manner. Serum aspartate aminotransferase, alanine aminotransferase, alkaline phosphatase, γ-glutamate transferase, urea, and creatinine (P < 0.05), and malondialdehyde concentrations in both serum and liver (P < 0.001) were also increased linearly in a ZEA-dose-dependent manner. However, spleen relative weight (P = 0.002) and activities of total superoxide dismutase and glutathione peroxidase (in both serum and liver (P < 0.05) were decreased linearly as dietary ZEA increased. Results showed that besides genital organs, the liver, kidney, and spleen may also be target tissues in young gilts fed diets containing 1.1 to 3.2 mg of ZEA/kg for 18 d. Increased key liver enzymes in the serum suggest progressive liver damage caused by feeding ZEA, and an increase in oxidative stress in gilts is another potential impact of ZEA toxicity in pigs.     

Influence of transport stress on serum cortisol and thyroid hormones in pigs with Halothane gene

The influence of transport stress on rectal temperature, respiratory rate and serum concentrations of cortisol, thyroxine and triiodothyronine (T3) in normal pigs (NN) and in heterozygous (Nn) pigs with Halothane gene (n), a major gene that causes the pale, soft, exudative meat (PSE), were examined in a hot environment (32–34°C). Easily releasable myofilament content of Longissimus dorsi taken by needle biopsy was also measured as an index of myofibrillar disassembly. Rectal temperature and respiratory rate were significantly increased by the transport. However, the influence was higher (P < 0.01) in Nn pigs. Serum concentration of cortisol was increased (P < 0.01) by the transport despite the genotypes. Serum concentration of T3 was not changed by the transport in Nn pigs, whereas in NN pigs it was significantly decreased after the transport (P < 0.01). Easily releasable myofilament content after the transport was higher in Nn pigs than that of NN pigs (P < 0.05). These results suggest that response of thyroid function might relate to the stimulation of muscle proteolysis and PSE in Nn pigs under stressful conditions.     

Effect of temperature on the sodium sulphite precipitation test for assessment of goat serum immunoglobulin

Total serum immunoglobulin concentrations of apparently normal indigenous goats were estimated by a sodium sulphite precipitation test which utilized 3 concentrations of the salt (i.e. 14%, 16%, 18%). A total of 210 goat serum samples comprising five phenotypically different breeds were examined. Immunoglobulin precipitation was greatly influenced by the incubation temperature. Using different concentrations of the sodium sulphite salt solution, serum samples incubated at either 4°C or 38°C consistently gave clear and rapid precipitation reaction in all samples with immunoglobulin concentrations of over 15 mg/ml.

Tests carried out at normal tropical room temperature (28°–30°C) gave inconsistent results and only 52% gave clear precipitation. Neither breed nor sex had any statistically significant effect on either the precipitation rate or the immunoglobulin values (P < 0.05).

It was concluded that this test if performed at either 4°C or 38°C can be used under field conditions to evaluate immune status of neonatal goats in the tropics.     

Relationships among serum immunoglobulin concentration in foals, colostral specific gravity, and colostral immunoglobulin concentration

Postpartum, presuckle, colostrum samples were collected from 100 mares. Colostral specific gravities significantly correlated (r = 0.9) with colostral immunoglobulin (Ig)G concentrations. Foal serum IgG concentrations highly correlated (r = 0.82) with specific gravities of the colostrum each foal ingested. Eight of 48 foals (17%) had serum IgG concentrations less than 400 mg/dl. The dams of these 8 foals had colostral sp gr less than 1.06 and colostral IgG concentrations less than 3,000 mg/dl. Foals had serum IgG concentrations greater than 520 mg/dl 24 hours after parturition, when the colostral specific gravity of the dam was greater than or equal to 1.06. Effects of breed on colostral specific gravity, colostral IgG concentrations, foal serum IgG concentrations, and mare serum IgG concentrations were not significant.     

Elimination of sulfamethazine from edible tissues, blood, urine, and feces of turkey poults.

Sulfamethazine was administered to 8- to 10-week-old turkey poults intravenously (IV) at the dose level of 71.5 mg/kg of body weight, orally at the dose level of 143 mg/kg of body weight, or in the drinking water at the concentration of 0.1% over a 6-day period. The concentrations of free sulfamethazine in blood, muscle, skin, kidney, and liver were determined and semilogarithmic plots of concentration vs time for the various tissues indicated that the curve had a linear portion within the first 72-hour period of drug withdrawal. The rates of disappearance of sulfamethazine from the various tissues were proportional to the concentration in the tissues. After 72 hours of withdrawal and for as long as 14 days, sulfamethazine concentrations in kidney, liver, and skin of turkeys given the drug in the drinking water fluctuated between 0.1 and 0.4 ppm. Only 8.6% of the oral dose (143 mg/kg) and 16.5 to 17% of the IV dose (71.5 mg/kg) were recovered in urine and feces as the parent compound during the initial 72-hour period.     

Effects of transportations, a high lactose diet and ACTH injections on the white blood cell count, serum cortisol and immunoglobulin G in young calves

Twelve calves were subjected to 5 different, consecutive treatments considered to induce stress. These included a) transportations for 4–8 h at temperatures just above or well below 0°G at the start of the experiment and 6 and 11 weeks later, b) feeding a high lactose diet and c) ACTH injections.The transportations resulted in a transient suppression in the level of serum IgG, and increase of Cortisol, blood neutrophils and lymphocytes. One and 2 weeks after the first transportation, the lymphocyte count was suppressed.Intramuscular injections of ACTH also resulted in temporary increases in serum Cortisol, blood neutrophils and lymphocytes, but no obvious changes in IgG. Thus, the suppressed levels of IgG which were seen after the transportations did not appear to be directly related to the increased level of Cortisol.The high lactose diet was not accompanied by a significantly higher frequency of diarrhoea and lower mean weight gain. The IgG level did not appear to be influenced by the lactose level of the diet, and calves fed a high lactose diet responded similarly to transportation as calves fed a normal type of milk replacer. Apart from some diarrhoea, no health problem occurred.     

Curcumin attenuates the effects of transport stress on serum cortisol concentration, hippocampal NO production, and BDNF expression in the pig

Curcumin, the active component of curcuma longa, has been reported to be effective in alleviating chronic stress-induced disorders in rodents by modulating neuroprotection and neuroendocrine functions of the central nervous system, especially hippocampus. However, it is unclear whether curcumin can attenuate the subacute stress response induced by 2 h of road transport in the pig. Therefore, the present study was designed to identify the changes of serum cortisol concentration, hippocampal nitric oxide (NO) production, and related gene expression in response to 2 h of transport and to explore whether curcumin treatment (8 mg/kg, p.o.) for 21 d before transport may alleviate the stress-induced responses in the hippocampus of pigs. We found that 2 h of transport elevated serum cortisol concentration (P < 0.01), increased hippocampal NO content, and reduced brain-derived neurotrophic factor (BDNF) mRNA expression in pigs not treated with curcumin, whereas these stress responses were all reversed or attenuated in curcumin-treated pigs. In addition, the stress-induced increase in the expression of constitutive nitric oxide synthase (cNOS) and enzyme activities of total NOS, cNOS, and inducible NOS (iNOS) was also reversed or attenuated in curcumin-treated pigs. However, neither transport nor curcumin caused significant alterations in hippocampal expression of 11β-hydroxysteroid dehydrogenase type 1 and type 2 (11β-HSD1 and 2), glucocorticoid and mineralocorticoid receptors (GR and MR), or pro-/anti-apoptotic molecules (Bax-α and Bcl-xL). These results suggest that curcumin can alleviate subacute stress response in pigs through its neuroprotective effects on modulating hippocampal NO production and BDNF expression.     

Study on cortisol,cortisone and prednisolone presence in urine of Chianina cattle breed

The Chianina, one of the oldest and most important cattle breeds of Italy, is now reared all over the world. The Chianina has been known and appreciated since ancient times because, from a nutritional point of view, its meat has no proper rivals. To date, studies have been performed to evaluate the genetic profile of the breed, but knowledge about the chemical profile is generally lacking. Due to the increased interest from farmers regarding breeding of the Chianina, this study proposes a preliminary evaluation of main endogenous urinary corticosteroids (cortisol and cortisone) and most commonly used synthetic one (dexamethasone). Moreover, after recent findings regarding the presence of endogenous prednisolone in the urine of more popular breeds, particular attention was given to analysis of the presence of prednisolone and prednisone, as well. For this aim, the urine samples of 12 young cows and 30 young bulls was collected at the farms and analysed using a fit‐for‐purpose LC‐MS/MS method. The preliminary results of this study show that prednisolone was found only in Chianina females (3 out of 12). Cortisol and cortisone were found at concentrations that showed a high inter‐individual variability, and that were higher in female urine compared to that of males.     

姜黄色素缓解猪运输应激作用的研究

姜黄色素是姜黄的主要活性成分,它也是中国传统中药逍遥散和解郁散的主要成分。据报道,姜黄色素具有潜在的抗氧化（Nishinaka等,2007）、抗炎（Jurenka,2009）、抗癌的作用（Villegas等,2008）。     

Vitamin A deficiency: serum cortisol and humoral immunity in lambs

Serum cortisol and antigen-specific and polyclonal immunoglobulin G (IgG) concentrations were measured to investigate the relationship between vitamin A status and immune function in lambs. Twenty-four 3-mo-old crossbred ewe lambs weighing approximately 10 kg were each fed 900 g/d of a carotene-deficient diet. The 12 control lambs also received a 100,000 IU oral dose of vitamin A palmitate every 2 wk. All lambs were given primary and secondary antigenic challenges. Lambs were slaughtered at the end of the secondary challenge period. Liver vitamin A concentrations were greater (P less than .001) in the control animals (69.5 vs 1.3 micrograms/g wet tissue). Both groups of lambs exhibited a similar growth response until d 105, after which daily gain of the control lambs exceeded (P less than .03) that of the A-deficient lambs. Polyclonal serum IgG concentrations were greater (P less than .05) in the A-deficient lambs on d 49 to 124 and on d 151 (P less than .10). Ovalbumin-specific serum IgG concentrations tended to be greater in the control lambs throughout the primary and secondary challenge periods. Control lambs had greater titers on d 164 (P less than .07) and d 190 (P less than .03). Vitamin A status appeared to have no consistent effects on serum cortisol concentrations. Spleen weights were greater (P less than .002) in the A-deficient lambs. Lungs from 11 of 12 A-deficient lambs contained abscesses, as opposed to 1 of 12 for the control lambs. Both polyclonal and antigen-specific IgG concentrations were affected by vitamin A status. Serum cortisol concentrations did not appear to mediate this effect.