Correlation of ribonuclease zymograms and incompatibility genotypes in almond

Proteins were extracted from styles of 29 self-incompatible cultivars of almond and separated using non-equilibrium pH gradient electro-focusing, and the gels were stained for ribonuclease activity. Mutually incompatible cultivars had similar banding patterns and, for the 24 cultivars already genotyped in France or California, the bands correlated well with the reported alleles. The band corresponding to S₁ of the French labelling system was indistinguishable from that corresponding to S_b of the Californian labelling system, and a controlled cross confirmed that these alleles are identical. The band corresponding to the Californian S_a was distinct from the bands corresponding to French alleles and, to harmonise the allele labels, it was redesignated S₅. The genotypes of five uncharacterised self-incompatible cultivars were inferred from zymograms as follows: ‘Desmayo Largueta’ and ‘Glorieta’, S₁S₅, ‘Masbovera’, S₁S₉, ‘Tarragones’, S₂S₉, and ‘Tokyo’, S₆S₇. The alleles designated S₆ and S₉ have not previously been reported. Nine self-compatible cultivars or selections were analysed, and each showed a band corresponding to an incompatibility allele as well as a common band; however, the correspondence of this common band to S_f, the allele for self-compatibility, is unproven. This revised version was published online in July 2006 with corrections to the Cover Date.     

Inheritance of stylar ribonucleases in cherry progenies, and reassignment of incompatibility alleles to two incompatibility groups

Stylar proteins were extracted from parents and seedlings of six progenies of cherry (Prunus avium), separated using isoelectric focusing, and the gels stained for ribonuclease activity. The zymogram of each plant showed two main ribonuclease bands in the region pI 8.3 to 9.6. Progenies from crosses of parents with one band in common segregated into just two classes, whereas progenies from crosses of parents with no common bands segregated into four classes, the two types of segregation corresponding to those expected from semi-compatible and fully-compatible crosses respectively. This behaviour was consistent either with the ribonuclease locus being tightly linked with the self-incompatibility, S, locus, or else with the S locus coding for the ribonuclease variants. Evidence favouring the latter hypothesis is discussed. An apparently anomalous segregation led us to assign to ‘Bradbourne Black’ a genotype different from that previously reported, and analysis of some other cultivars in the same incompatibility group, Group VII, led us to conclude the genotype of this group is S₃S₅, and not S₄S₅ as previously reported. Correspondingly, we suggest the genotype of Group V is S₄S₅, and not S₃S₅. Five new S alleles, S₇, S₈, S₉, S₁₀ and S₁₁ were proposed in parental cultivars and selections that had not previously been assigned a genotype. This revised version was published online in July 2006 with corrections to the Cover Date.     

Comparison of isoenzymes in Prunus avium separated by two different electrophoretic techniques

Isoenzyme variation for seven systems revealed by two different electrophoretic procedures was compared in Prunus avium. Fourteen cultivars and 14 wild selections were analysed for acid phosphatase (ACP), isocitrate dehydrogenase (IDH), leucine aminopeptidase (LAP), malate dehydrogenase (MDH), phosphoglucomutase (PGM), shikimate dehydrogenase (SKD) and superoxide dismutase (SOD). Extracts were separated by isoelectric focusing (IEF) and by polyacrylamide gel electrophoresis (PAGE). For the eight loci that had been described previously in these enzyme systems on the basis of IEF analysis, we compared the variation revealed with IEF and PAGE. Similar variation was revealed for Acp‐1 and Pgm‐1, and the alleles revealed by PAGE could be identified directly with those reported for IEF. For Lap‐1, Mdh‐1 and Skd‐1, variation was seen with IEF but not with PAGE. For Mdh‐2, PAGE revealed additional variation not revealed by IEF. For Idh‐1, different patterns of variation were revealed by PAGE and IEF, and both procedures would be needed to genotype cherry accessions. We were unable to detect variation corresponding to that reported previously for Sod‐1 with either technique. The implications of these findings for allele labelling, for studies of genetic diversity and for linkage analysis are discussed.     

A gene for susceptibility to the fungicide azoxystrobin in apple and a tightly linked microsatellite marker

Various apple cultivars, including 'McIntosh' and 'Cox', are reported to be susceptible to the strobilurin fungicide azoxystrobin whereas others, including 'Delicious' and 'Golden Delicious', are resistant. To investigate the genetic control, progenies from various crosses between these four cultivars were raised and the seedlings tested for response by painting three expanded leaves with 200 mg/l of azoxystrobin solution and noting symptoms about a week later. From the segregations, it was concluded that susceptibility to azoxystrobin is due to a dominant gene, Azs , for which 'McIntosh' and 'Cox' are heterozygous. By scoring the segregation in the mapping progeny 'Fiesta' × 'Totem' and comparing it with the segregation of microsatellite [simple sequence repeat (SSR)] markers, Azs was found to co-segregate with SSR-GD127 on linkage group 12 and this tight linkage was confirmed in the progeny from the cross of 'Golden Delicious' × 'Cox' and 'McIntosh'   ×   'Golden Delicious'. The susceptibility of 'McIntosh', attributed to allele Azs-m , was greater than that of 'Cox', Azs-c , and in the 'McIntosh'   ×   'Cox' progeny the GD127 marker showed that Azs-c is dominant to Azs-m .     

Ostertagiasis in calves. II. infection parameters and body weight gains following housing

The performance of six groups of six calves each from a previous field experiment was followed from the beginning of housing until turning out seven months later. When housed the groups harboured different levels of infection with Ostertagia ostertagi and had corresponding weight and gain reductions. The most heavily infected groups improved rapidly following housing. Elevated serum pepsinogen levels decreased markedly, and approached normal values at eight weeks. Differences in serum pepsinogen levels between groups diminished considerably, but remained significant over the entire stabling period. The serum albumin level was low in the most seriously affected animals at the time of housing, but normal levels were restored within the first twelve weeks. In general, the number of parasite eggs in the faeces of the animals decreased, but there was considerable fluctuation. Apart from a single sampling date, no significant difference in egg per gram (EPG) could be demonstrated between the experimental groups during the stabling period.The calves were fed according to their age. The live weight differences between most and least affected groups diminished from 64 kg at the time of housing to 37 kg at the end of the stabling period. The reduction took place particularly during the first four weeks of housing.Considering both the grazing and stabling periods, it appeared that anthelmintic treatment twice during the period had no effect on the final live weight, whereas remeated treatments at three-week intervals resulted in an increase of 24 kg. Similarly, moving the animals to a clean pasture in mid-July resulted in an increase of 39 kg at the end of the stabling period when compared to calves which were not moved. Treatment of moved animals did not result in further body weight gain after seven months of housing.No significant correlation was found between parasite EPG of faeces and growth rate during the stabling period. However, a positive correlation was found between serum pepsinogen during the first eight weeks of housing and the weight gain over the entire stabling period. This was in contrast to the correlation experienced during the grazing period.     

Comparison of opioid and alpha-2 adrenergic receptor location and density in the horse and dog using radioligand binding

Ketamine, a noncompetitive NMDA receptor antagonist, has been shown to provide analgesia in some species. To target the NMDA receptor specifically and to potentially minimize some untoward side-effects, ketamine had been used epidurally. The objective of this study was to determine the analgesic effect of epidurally administered ketamine in dogs with chemically induced synovitis.
Sixteen healthy dogs were used. Dogs were anesthetized with propofol (4 mg kg⁻¹ IV). Synovitis was induced by injecting 1 mL of sodium urate crystal solution (10 mg mL⁻¹) into the right stifle. Dogs were allowed to recover and the synovitis was allowed to develop for 12 hours. The dogs were then anesthetized again using propofol (4 mg kg⁻¹ IV). Lumbosacral epidural injections were performed with each dog receiving either 2 mg kg⁻¹ of ketamine (20 mg mL⁻¹) or an equal volume of placebo (sterile water containing not more than 0.1 mg mL⁻¹ benzethonium chloride). Analgesia was assessed at baseline and then at 12, 14, 16, 18, 20, and 24 hours after induction of synovitis. Ground reaction forces (peak vertical force and impulse area) and overall pain were measured using a force platform and a pain scoring system (numerical rating scale).
Analysis of the data by Repeated Measures anova showed that the dogs developed a significant lameness between the baseline and 12 hours. However, no significant difference in ground reaction forces or total pain score was demonstrated between the treatment and control groups at any other time.
In conclusion, ketamine administered epidurally at a dose of 2 mg kg⁻¹ did not provide significant analgesia in dogs with chemically induced synovitis.     

A Screening for the Occurrence of Autoreactive Antibodies in Sera of Pregnant and Non-pregnant Bitches

Sera of healthy pregnant (group I, n = 11) and non-pregnant (group II, n = 11) bitches were screened for autoantibodies (AAb). In both groups, blood samples were drawn every fifth day between days 5 and 55 after mating. Serum was analysed via indirect immunofluorescence (IIF) with the Canine ANA HEp-2 Screening Kit. In all animals, anticytoplasmic AAb were detected. Utilizing primate-heart substrate slides AAb against contractile proteins of the cytoplasm could be observed. The predominating fluorescence pattern in pregnant animals resembled above all desmin, which was proven via Western blot. The sera were then pre-incubated with tropomyosin, actin, vimentin, vinculin and keratin solutions, and assessed on HEp-2 slides and on human and canine fibroblasts as well. The latter substrate was used to verify whether the detected Ab were in fact AAb. Utilizing tropomyosin, revealed elimination of the cytoplasmic fluorescences on all three substrates. It is therefore assumed, that in sera of healthy dogs, AAb against contractile structure proteins of the cytoplasm are present regularly. The majority of pregnant bitches presented with higher end-point titres (EPT), than to be found in non-pregnant dogs. AAb against desmin played the key role in those patterns. In addition, sera were screened for thyroid specific AAb, namely thyroglobulin, thyroid peroxidase (TPO), T3 and T4, and for AAb against insulin by ELISA or Western blot (TPO). Only in two of the pregnant bitches a weak positive reaction (1:100) for T3-AAb was detected.     

Associations between hoof shape and the position of the frontal plane ground reaction force vector in walking horses

AIMS: To determine the frontal plane position of the ground reaction force vector at its centre of pressure under the hoof of walking horses, and its projection through the distal limb joints, and to relate this to hoof geometric measurements.

METHODS: Reflective markers were glued to the forelimb hooves and skin of 26 horses, over palpable landmarks representing centres of the coffin, fetlock and carpal joints, and the dorsal toe at its most distal point. A 4-camera kinematic system recorded the position of these markers as the horse walked in hand across a force platform, to generate a frontal plane representation of the ground reaction force vector passing between the markers at the joints. The position of the vector was calculated as the relative distance between the lateral (0%) and medial (100%) markers at each joint. Digital photos were taken of the hoof in frontal and sagittal views to determine hoof geometric measurements. Associations between these and the position of the force vector at each joint were examined using Pearson correlation coefficients.

RESULTS: Mean vector position for both forelimbs at the toe, coffin, fetlock and carpal joint was 50.1 (SD 8.9), 53.0 (SD 9.2), 54.6 (SD 11.4) and 50.5 (SD17.3)%, respectively, of the distance between the lateral and medial sides of the joint in the frontal plane. Across all four joints, the vector position was slightly more medial (2–4%) for the right than left limb (p>0.05). Medial hoof wall angle was correlated (p<0.05) with force vector position at the fetlock (r=?0.402) and carpal (r=?0.317) joints; lateral hoof wall angle with vector position at the toe (r=0.288) and carpal (r=?0.34) joint, and medial hoof wall height with vector position at the fetlock (r=?0.306) and carpal (r=?0.303) joints.

CONCLUSION: The position of the two-dimensional frontal plane ground reaction force vector at the toe, and at the fetlock and carpal joints was associated with hoof shape. Mediolateral hoof balance has been shown in vitro to affect articular forces, which may be a factor in development of joint disease. The effect of hoof shape needs to be evaluated at faster gaits to determine the potential for joint injury in the presence of larger forces.     

Sacral osteochondrosis in two German Shepherd Dogs

Two young adult male castrated German Shepherd Dogs were referred for evaluation of intermittent episodes of hindlimb pain. Physical examination suggested lumbosacral stenosis, and plain radiographs and computed tomography revealed lesions consistent with sacral osteochondrosis. One dog had osteochondral fragments removed surgically; the other was managed conservatively. The surgically treated dog had complete resolution of clinical signs whereas the dog managed conservatively had repeated episodes of mild pain and received one short course of non-steroidal anti-inflammatory medication in 18 months. Sacral osteochondrosis has not been previously reported in Australia.     

微量Bradford法测定提纯禽结核菌素蛋白含量

研究用96孔微量板Bradford法测定提纯禽结核菌素蛋白含量。通过不同浓度的待测禽结核菌素与考马斯亮蓝G-250溶液混合后，在595nm光波下测定各孔样品的OD值。微量Bradford法检测禽结核菌素的线性范围是50—1000μg／mL，相关系数为0．999，最低定量限为50μg／mL。试验用3支禽结核菌素国际参照品，分别配制成125、250、500μg／mL三种不同浓度，每支每种浓度测定4次，计算实际测得的蛋白质浓度。结果显示，批内相对标准偏差为1．6％～4．4％；批间相对标准偏差0．7％-2．2％。研究结果表明微量Bradford法可作为测定禽结核菌素蛋白含量的一种方法。