Skeletal muscle fiber size in untrained and endurance-trained horses.

The mean area and minimal diameter of 3 histochemically determined myofiber types (1, 2A, and 2B; myosin ATPase in acid buffer) were calculated in middle gluteal muscle biopsy specimens from 62 stallions, 47 Andalusians and 15 Arabians, ranging in age from 6 to 12 years. Fourteen Andalusians and 7 Arabians were untrained, and the remainder were actively endurance-trained. The 6-month training schedules involved walking, slow trotting, and cantering. Fourteen Andalusians were moderately endurance-trained, whereas the other 19 Andalusians and 8 Arabians were strongly endurance-trained. Significant differences were not recorded between untrained and endurance-trained Arabians with respect to the area (type 1, 3,194 +/- 869 microns 2 and 3,150 +/- 370 microns 2; type 2A, 3,819 +/- 890 microns 2 and 3,380 +/- 356 microns 2; and type 2B, 4,872 +/- 962 microns 2 and 4,417 +/- 646 microns 2) or minimal diameter (type 1, 52.2 +/- 7.4 microns and 52.8 +/- 3.1 microns; type 2A, 58.1 +/- 6.7 microns and 55.0 +/- 2.8 microns; and type 2B, 65.3 +/- 6.4 microns and 63.4 +/- 4.3 microns) of the 3 fiber types, nor between untrained and endurance-trained Andalusians with respect to the area (untrained, 3,990 +/- 690 microns 2; moderately endurance-trained, 3,882 +/- 347 microns 2; and strongly endurance-trained, 3,758 +/- 510 microns 2) and minimal diameter (untrained, 58.1 +/- 4.7 microns; moderately endurance-trained, 59.7 +/- 2.7 microns; and strongly endurance-trained, 58.7 +/- 4.5 microns) of 2A fibers.(ABSTRACT TRUNCATED AT 250 WORDS)     

Physical characteristics and chemical composition of Greater Rhea (Rhea americana) eggs from wild and captive populations.

1. Eggs from wild and captive populations of Greater Rhea (Rhea americana) were studied to determine their physical and chemical characteristics. 2. Significant differences were found among populations in almost all chemical parameters studied, whereas within physical parameters only shell weight (as a proportion of the entire egg) and density showed differences. 3. Eggs from wild populations had the highest protein and linolenic acid and the lowest total lipid contents, while cholesterol levels of these eggs and of those from the largest captive area were the lowest.     

Determination of sulfamethazine and trimethoprim in liquid feed premixes by HPLC and diode array detection, with an analysis of the uncertainty of the analytical results.

Sulfamethazine (SMZ) and trimethoprim (TMP) are antibacterials used in veterinary practice. This paper describes a method for their determination in veterinary liquid feed premixes that is based on liquid chromatography with diode array detection. Gradient elution with methanol and ammonium acetate achieved excellent separation of the two analytes within 15 min without any interference from the matrix. Absorbance of the column effluent was monitored at 264 nm for SMZ and at 230 nm for TMP. Detailed analyses of the uncertainties of determinations afford estimated expanded uncertainties of, respectively, 0.2 and 0.1 w/v % for typical SMZ and TMP concentrations of 10.7 and 2.1 w/v %, respectively. At the lower end of the calibrated range of the method, the dominant source of uncertainty is the preparation of standards and the construction of the calibration line.     

Biochemical polymorphisms in Spanish Avileña-Negra Iberica cattle

SUMMARY: Thirteen biochemical blood polymorphisms were analysed in a population of 149 Spanish Avile?a-Negra Ibérica cattle. The study revealed variation at the following nine loci: HBB, CA, NP, CP, AMY1, ALB, GC, TF and PTF2. The following systems were monomorphic: CAT, DIA1, MDH1 and ME1. Using polyacrylamide-gel electrophoresis, a slow, migrating pair of bands was found in the GC protein system. This pattern is probably controlled by the GC(C) allele, described in only a few cases in cattle. Furthermore, starch-gel electrophoresis allowed the detection of a variant with intermediate mobility between the ALB(A) and the ALB(B) alleles at the albumin locus. A variant with a similar electrophoretic pattern has occasionally been reported in cattle. However, utilizing IEF under denaturing conditions, such a variant could not be differentiated from the ALB(A) allele and thus its significance is not clear. ZUSAMMENFASSUNG: Biochemischer Polymorphismus in spanischen Avile?a-Negra Iberica Rindern Insgesamt 13 biochemische Systeme wurden in einer Population von 149 spanischen Avilena-Negra-Iberika-Rindern hinsichtlich Polymorphismus analysiert. Es zeigten sich Varianten bei folgenden Loci: HBB, CA, NP, CP, AMY1, ANB, GC, TF und BTF2, w?hrend CAT, DEA, MDH1 und ME1 monomorph sind. Bei st?rke Gel-Elektrophorese wurde im Albumin-Locus eine Variante mit intermedi?rer Mobilit?t zwischen ALB(A) und ?LB(B) Allel entdeckt. Eine solche Variante wurde bisher nur sehr selten bei Rindern beobachtet. Darüber hinaus wurde bei Polyacrylamid-Gel-Elektrophorese ein langsam wanderndes Paar von B?ndern im GC-Proteinsystem gefunden. Dieses Muster ist wahrscheinlich von dem selten vorkommenden GC(C) -Allel verursacht. RESUMEN: Se analizaron trece polimorfismos bioquímicos sanguíneos en una población de 149 animales de la raza Avile?a-Negra Ibérica de ganado vacuno. El estudio reveló la existencia de variación en los nueve loci siguientes: HBB, CA, NP, CP, AMY1, ALB, GC, TF y PTF2. Fueron monomórficos los sistemas siguientes: CAT, DIA1, MDH1 y ME1. Utilizando eletroforesis en gel de poliacrilamida se encontró un par de bandas de migración lenta en el sistema de la proteína GC. Este patrón probablemente está controlado por el infrecuente alelo GC(C) , descrito en unos pocos casos en el ganado vacuno. Además, la electroforesis en gel de almidón permitió detectar en el locus de la albúmina una banda con movilidad intermedia entre los alelos ALB(A) y ALB(B) . Una variante con un patrón electroforético similar ha sido descrita en muy pocas ocasiones en el ganado vacuno. Sin embargo, la utilización de IEF en condiciones desnaturalizantes no permitió diferenciar esta variante del alelo ALB(A) y, por lo tanto, el significado de la misma no está claro.     

‘Plantibodies’: a flexible approach to design resistance against pathogens

Engineering resistance against various diseases and pests is hampered by the lack of suitable genes. To overcome this problem we started a research program aimed at obtaining resistance by transfecting plants with genes encoding monoclonal antibodies against pathogen specific proteins. The idea is that monoclonal antibodies will inhibit the biological activity of molecules that are essential for the pathogenesis. Potato cyst nematodes are chosen as a model and it is thought that monoclonal antibodies are able to block the function of the saliva proteins of this parasite. These proteins are, among others, responsible for the induction of multinucleate transfer cells upon which the nematode feeds. It is well documented that the ability of antibodies to bind molecules is sufficient to inactivate the function of an antigen and in view of the potential of animals to synthesize antibodies to almost any molecular structure, this strategy should be feasible for a wide range of diseases and pests.Antibodies have several desirable features with regard to protein engineering. The antibody (IgG) is a Y-shaped molecule, in which the domains forming the tips of the arms bind to antigen and those forming the stem are responsible for triggering effector functions (Fc fragments) that eliminate the antigen from the animal. Domains carrying the antigen-binding loops (Fv and Fab fragments) can be used separately from the Fc fragments without loss of affinity. The antigen-binding domains can also be endowed with new properties by fusing them to toxins or enzymes. Antibody engineering is also facilitated by the Polymerase Chain Reaction (PCR). A systematic comparison of the nucleotide sequence of more than 100 antibodies revealed that not only the 3′-ends, but also the 5′-ends of the antibody genes are relatively conserved. We were able to design a small set of primers with restriction sites for forced cloning, which allowed the amplification of genes encoding antibodies specific for the saliva proteins ofGlobodera rostochiensis. Complete heavy and light chain genes as well as single chain Fv fragments (scFv), in which the variable parts of the light (V_L) and heavy chain (V_H) are linked by a peptide, will be transferred to potato plants. A major challenge will be to establish a correct expression of the antibody genes with regard to three dimensional folding, assembly and intracellular location.     

C-reactive protein measurement in canine saliva.

An established time-resolved immunofluorometric assay designed for measurement of C-reactive protein (CRP) in canine blood was evaluated and validated for use in canine saliva. C-reactive protein was measured in saliva specimens from 5 healthy dogs before and after the injection of casein and in 37 dogs with different disease conditions. The analytical and functional limits of detection were 0.000053 microg/ml and 0.0091 microg/ml, respectively, and intra- and interassay coefficients of variation ranged between 6.7-9.9% and 8.5-16.5%, respectively. A recovery experiment showed no significant disagreement between detected values and expected ones, and saliva CRP concentration was measured in a linear and proportional manner. A positive correlation was found between CRP levels obtained in saliva and serum samples in the experimental (R2 = 0.76) and clinical studies (R2 = 0.70). The assay was able to detect significant differences between salivary CRP levels in healthy dogs and dogs with inflammatory processes. These results suggest that saliva can be used for CRP measurement in dogs. The use of saliva presents the advantage of an easier and less stressful sampling method for the animals, which might be performed outside of hospital environments.     

Characteristics for Practical Use of Attenuated Isolate UA-Fukushima of <Emphasis Type="Italic">Tomato mosaic virus</Emphasis>

L₁₁A-Fukushima (L₁₁A-F) derived from attenuated isolate LuA of Tomato mosaic virus (ToMV) has the highest ability to cross protect against virulent ToMV among LuA and its derivatives and is stably inherited. Growth, yield, fruit quality and symptom attenuation of inoculated tomato plants did not differ significantly between L₁₁A-F and L₁₁A. The infectivity of progeny viruses in tomato infected with LuA-F was less than 4% of that with virulent ToMV. From these results, L₁₁A-F appears to possess the properties necessary for practical use. To manage L₁₁A-F strictly, a PCR-based assay to detect trace contamination of virulent ToMV in L₁₁A-F preparations was established. Received 10 June 2002/ Accepted in revised form 30 October 2002