生物素测定方法的应用现状

<正>生物素,又称维生素H,是一种水溶性B族维生素。生物素的测定方法很多,早在20世纪40年代,国外就报道了利用微生物法测定生物素的含量。随着科学技术的发展,国内外报道了许多新的测定方法,主要有同位素稀释法、     

Incidence of post-anaesthetic arrhythmias in dogs

OBJECTIVES: To assess the use of Holter monitoring for evaluating the incidence of post-anaesthetic cardiac arrhythmias and associated anaesthetic risk for two different anaesthetic protocols. METHODS: Patients undergoing orthopaedic surgery were randomly divided into two groups with different anaesthetic regimens (group A, isoflurane n = 30; group B, propofol n = 30). Two 24-hour Holter recordings were performed for each patient: the first directly following anaesthesia and the second, as a comparison, on the fifth postoperative day. RESULTS: Although all dogs were healthy on pre-anaesthetic cardiac evaluation, 56 dogs showed arrhythmias in the two 24-hour (Holter) electrocardiograms performed. However, the number of arrhythmias recorded was low in most cases (less than 10 supraventricular extrasystoles and less than 100 ventricular extrasystoles). One patient in group A showed 94 supraventricular extrasystoles during the second monitoring period. Three patients in each group developed more than 100 ventricular extrasystoles during both Holter recordings. There were no statistically significant differences between the two anaesthetic regimens or between the two recordings in both groups. CLINICAL SIGNIFICANCE: The two anaesthetic protocols investigated in this study did not induce an increased incidence of severe arrhythmias in healthy dogs in the post-anaesthetic phase.     

Retrospective study of bacterial infective arthritis in 31 dogs

OBJECTIVES: To characterise the presenting signs and pathological changes of canine bacterial infective arthritis in 31 dogs, and to document the response to different treatment regimens. Risk factors that may predispose joints to bacterial infective arthritis and influence the success of treatment were also investigated. METHODS: A retrospective review of cases of bacterial infective arthritis that were presented to three university veterinary referral hospitals over a five-year period (January 1997 to January 2002) was performed. RESULTS: The elbow joint (38 per cent) and stifle joint (44 per cent) were most commonly affected. Radiographic changes consistent with pre-existing osteoarthritis were identified in 14 joints, which had no history of previous surgery (articular or periarticular) or penetrating wound. No significant difference (P = 0.78) was identified between the outcome of combined surgical and medical management, and medical management alone. There were trends for poorer outcomes with increased bodyweight of the dog, longer duration of lameness and a higher nucleated cell count of the affected joint fluid at presentation. The overall infection rate for articular surgical procedures at one institution was 1-3 per cent. CLINICAL SIGNIFICANCE: Medical and/or surgical management were usually successful in resolving infection (94 per cent). However, they were frequently unsuccessful in restoring full joint function; this may in part have been due to the nature of the underlying joint     

A novel subgroup among genotypes of equine arteritis virus: genetic comparison of 40 strains

The authors determined partial nucleic sequences of the variable regions of open-reading frame (ORF5) from 151 nucleotide to 668 nucleotide and deduced amino acid sequences of 518 nucleotide respectively of 20 equine arteritis virus (EAV) isolates. About 19 Hungarian and one Austrian EAV strains were subjected to sequence analysis, the further data of 20 EAV strains: six North American and 14 European were obtained from the GenBank. Comparative sequence analysis of the Hungarian EAV strains indicated that among the three variable regions the first has been affected mostly by point mutations. Genetic comparison of the Hungarian strains with other EAV isolates from western Europe and North America (including the Bucyrus reference strain) has been performed on the aforementioned genome region. Besides the already known genetic subgroups of EAV; phylogenetic analysis revealed a novel subgroup comprising mainly Hungarian strains. Compared with the Bucyrus virus, the overall sequence divergencies of the examined Hungarian strains ranged from 81.47 to 90.73% at nucleotide and from 84.88 to 91.86% at amino acid level. Epizootiological studies have shown that the significant part of the EAV strains having been existed in Hungary before and in 2000 belong to this unique cluster (II.D) which was not indicated in former phylogenetic studies. After 2000 new EAV strains emerged in Hungary, one of them causing abortions or neonatal death. The previously dominant 'Hungarian' EAV genotypes were replaced by these new strains belonging to North American and European subgroups (I.A, I.B, II.A, II.B). The anamnesis of these cases revealed connections with persistent virus shedder stallions, those were imported to the country after 2000 or have been infected abroad. One of these Hungarian stallions became the source of abortion storms in Hungarian studs.     

The pharmacokinetics of vedaprofen and its enantiomers in dogs after single and multiple dosing

Vedaprofen is a chiral nonsteroidal anti-inflammatory drug that has been developed as a gel formulation for oral administration to dogs and horses. The pharmacokinetics of vedaprofen and its enantiomers were studied in beagle dogs after single (intravenous solution and oral gel) and multiple (oral gel) dosing at a dosage of 0.5 mg/kg body weight. Plasma concentrations of vedaprofen and its enantiomers were analysed by HPLC. The plasma protein binding of vedaprofen was studied by ultrafiltration. The absorption of vedaprofen was rapid (tmax 0.63 +/- 0.14 h) and almost complete after oral administration (bioavailability 86 +/- 7%). The terminal half-lives after intravenous and oral administration, 16.8 +/- 2.2 and 12.7 +/- 1.7 h respectively, were of the same order of magnitude. Enantioselective analysis showed that the R(-) enantiomer predominated in plasma. The change in the plasma time course of the plasma R(-)/S(+) enantiomer concentration ratio over time was similar after single intravenous and oral dosing, with R(-)/S(+) ratios in the AUC of 1.7 +/- 0.5 and 1.9 +/- 0.2 respectively. Plasma protein binding of vedaprofen and its enantiomers was high (> 99.5%). Vedaprofen is absorbed rapidly from the gastrointestinal tract, has a high bioavailability and does not accumulate in plasma in dogs following repeated oral administration.     

A comparison of the effects of feeding treatments and lighting on age at first egg and subsequent laying performance and carcase composition of broiler breeder hens

The effects of the growth curve from 15 to 20 weeks, age at photostimulation and pattern of photostimulus on sexual maturity, egg production and egg weight were evaluated in two trials with broiler breeder females to 56 and 34 weeks of age (housed in litter pens and individually caged, respectively). Carcase composition and reproductive morphology of hens varying in laying efficiency were measured in the second trial. Trial 1: Four growth curves were applied from 15 to 20 weeks to pullets housed in litter pens. Birds were transferred from 8- to 10-h photoperiods at 20 or 24 weeks of age, followed by weekly increments of one hour to reach a 16-h final photoperiod. Lower-than-recommended body weights at 20 weeks significantly delayed sexual maturity, reduced peak rate of lay, total eggs and mean egg weight. However, double-yolked egg production was lower, resulting in non-significant differences in settable egg numbers between body weight treatments. Birds reared to the heaviest body weight exhibited a significantly advanced sexual maturity, but total egg numbers, peak rate of lay and mean egg weight were not significantly affected. However, the laying of more double-yolked eggs resulted in a decrease in the number of settable eggs. Delaying photostimulation to 24 weeks significantly retarded sexual maturity, reduced total and settable egg numbers, and increased mean egg weight. A tendency for fewer double-yolked eggs was observed. Trial 2: At 19 weeks, birds were selected from the 4 body weight categories in Trial 1 and moved to individual cages in 8 rooms. Five lighting programmes were applied. The pattern of photostimulation applied did not affect any of the production traits measured. At 34 weeks, 24 birds were selected for the analysis of reproductive morphology, presence of internal or multiple ovulations, and carcase composition. Eight hens showing an early age at first egg and regular egg production records, 8 birds showing erratic laying performance and 8 non-layers were killed for this purpose. There were no differences in carcase composition or reproductive morphology between these groups. No internal ovulations or double or multiple ovarian hierarchies were observed. The results presented confirm that broiler breeders do not require a lighting stimulus in order to initiate ovarian activity and that, where no lighting stimulus is given, body weight or feeding level plays a critical role in stimulating the birds to attain sexual maturity. However, when a lighting stimulus is given, factors such as body weight and body composition become relatively less important in regulating the age at sexual maturity.     

Lymphocytes of dogs immunised with purified excreted-secreted antigens of Leishmania infantum co-incubated with Leishmania infected macrophages produce IFN gamma resulting in nitric oxide-mediated amastigote apoptosis

The role of nitric oxide (NO) in the anti-leishmanial activity has been confirmed both in vitro and in vivo. Recently, we demonstrated that NO-mediated apoptosis-like amastigote death pathway is an important and highly regulated mechanism used for the clearance of Leishmania within infected murine macrophages stimulated to produce NO endogenously. To further characterize these important effector mechanisms in dog, a natural host-reservoir of L. infantum/L. chagasi, we have developed an ex vivo infection model of canine macrophages. Exposure of L. infantum-infected macrophages to autologous peripheral lymphocytes derived from dogs immunised with purified excreted-secreted antigens of L. infantum promastigotes (LiESAp) formulated with muramyl dipeptide (MDP) as adjuvant resulted in a significant leishmanicidal effect due to interferon (IFN)-gamma dependent macrophage activation. Concomitant accumulation of NO(3)(-)/NO(2)(-) in supernatants of co-cultured cells and in situ staining of parasites with terminal deoxynucleotidyltransferase-mediated dUTP nick-end labelling (TUNEL) and YOPRO-1 showed that NO-mediated apoptosis of intracellular L. infantum amastigotes is occurring in canine macrophages as previously observed in mouse models. Monitoring these parameters in dogs after immunisation and before experimental challenge can represent a useful and easy way to rapidly evaluate vaccine candidates against canine visceral leishmaniasis.     

Porcine adenovirus as a delivery system for swine vaccines and immunotherapeutics

Porcine adenovirus (PAdV) has many qualities which make it an ideal choice for use as a delivery vector in swine. It is a low grade pathogen, present almost world-wide in a number of serotypes varying in their virulence and tissue tropism, which may allow for serotype specific vaccine targeting. PAdV is species specific having only been isolated from swine, reducing the possibility of its spread to other animals or man following administration. When engineered to contain a foreign gene, recombinant PAdV (rPAdV) can be grown to high titres in tissue culture cells making it cheap to produce. Knowledge of the complete nucleotide sequence of the PAdV genome has enabled rationally directed insertions of foreign genes which remain stably inserted in the genome and can be expressed at high levels following delivery to the target host. Importantly, recombinant PAdV can be administered by injection or by the oral route in feed or drinking water. We have delivered a range of antigens and immunomodulatory molecules to commercially available pigs using rPAdV and found it to be a very effective delivery system. Significantly, recombinant PAdV serotype 3 is highly effective as a delivery vehicle even when administered in the face of high levels of artificially induced serotype specific neutralising antibody to the vector.     

Effects of dietary cation-anion difference on blood cortisol and ACTH levels in reproducing ewes

Formulation of rations to induce a compensated metabolic acidosis in the post-partum cow has proved a useful strategy for prevention of milk fever. Such acidification improves the ability of the animal to maintain calcium homeostasis by promoting the absorption of calcium from the intestine and mobilization of calcium from the bone. In humans, an acidogenic diet results in mild metabolic acidosis in association with a state of cortisol excess and this increase in plasma cortisol may increase bone catabolism. The objective of our experiment was to induce acidification by anionic salt supplementation and to study the effects of anionic supplementation on plasma cortisol and adrenocorticotropic hormone (ACTH) levels in sheep. Twenty-seven twin-bearing sheep were assigned to two experimental groups and a control group, depending on dietary cation-anion difference (DCAD) (+272.6, -88.9 and +164.5 mEq/kg DM respectively). Sheep assigned to each dietary treatment received their respective rations beginning 6 weeks pre-partum and continuing until 12 days post-partum. Anionic diet induced a non-respiratory systemic acidosis in association with a mild increase in plasma cortisol concentration without changes in plasma ACTH levels. Our data suggest that the mild hypercortisolism observed in sheep fed the anionic diet may not be an effector for bone resorption induced by anionic salts. A mild hypercortisolism of this magnitude may lead to osteoporosis but this might require many years of adrenal hypersecretion while anionic salts are only used during the last weeks of pregnancy.     

Hyperextension of fetlock joints in German Holstein and German Holstein-Limousin crossbred calves

In two different dairy farms six calves exhibiting bilateral flexion of the fetlock joints in front or rear legs were born in 2000 to 2002. Four of the affected calves from the same farm were crossbred between German Holstein cows with red and white coat colour and a bull of the breed Limousin. The other two affected calves born on another farm were purebred German Holsteins with a black and white coat colour. The tests for BVD virus antigen and antibodies were negative in all affected calves. Three of the calves showed a lower selen and a higher glutamate dehydrogenase concentration in the analyses of blood metabolites. Two crossbred calves showed a degeneration of the liver with a progressive periportal fibrosis in a histological examination. In one calf an edema of astrocytes of the central nervous system was seen. The analysis of the pedigrees revealed for the four crossbred calves the Limousin bull as common ancestor and the mothers of the calves as relatives. For the affected purebred German Holstein calves also a sire was identified as a common ancestor. The pedigrees support inheritance through a monogenic autosomal recessive locus or more recessive gene loci with variable expressivity. However, the analysis could not clarify whether different gene loci are responsible for the congenital anomalies observed in the calves from the two farms and thus, the observed anomalies may be different genetic entities. Obvious environmental reasons were not found.