Hog Cholera: I. Investigation of the Agar Double-Diffusion Precipitation Test for The Detection of the Virus in Swine Tissue

The agar double-diffusion precipitation test was investigated to detect hog cholera virus in splenic and pancreatic tissues of swine. Special attention was given to the conditions influencing the sensitivity and specificity of the test. These studies emphasize the strict techniques and methods required in the test in order to detect specific antigen — antibody reaction. Absorption studies performed on serum fractions prepared by DEAE cellulose chromatography and studied electrophoretically, indicated the specific reaction was given by fractions, migrating in the gamma-globulin region and was absorbed by infected but not by normal spleens. The sensitivity of the test is dependent to a great extent on the successful liberation of the viral antigen from the tissue.     

Hog Cholera: IV. Detection of the Virus in Tissue Culture Preparations by the Fluorescent Antibody Technique

The fluorescent-antibody technique was employed for detection of hog cholera virus in tissue cultures inoculated with spleens of infected animals. As controls, cultures were also inoculated with material from normal swine and from those infected with other agents. In the first series 71 of 73 infected spleens, or 97 per cent, were detected. There were no false positive reactions among the controls. Results obtained with the second series of pigs showed that spleens collected during advanced stages of the disease were more satisfactory specimens than those collected earlier during the high temperature phase of infection. Findings with the third series of older swine indicated that their spleens were less satisfactory as a source of virus than those from young pigs. Tissues from freshly killed animals provided better specimen material than those from animals which had died.     

On-line gas chromatography combustion/pyrolysis isotope ratio mass spectrometry (HRGC-C/P-IRMS) of pineapple (Ananas comosus L. Merr.) volatiles

By use of extracts prepared by liquid-liquid separation of the volatiles from self-prepared juices of pineapple fruits (Ananas comosus) (n = 14) as well as commercial pineapple recovery aromas/water phases (n = 3), on-line capillary gas chromatography-isotope ratio mass spectrometry was employed in the combustion (C) and the pyrolysis (P) modes (HRGC-C/P-IRMS) to determine the delta(13)C(VPDB) and delta(2)H(VSMOW) values of selected pineapple flavor constituents. In addition to methyl 2-methylbutanoate 1, ethyl 2-methylbutanoate 2, methyl hexanoate 3, ethyl hexanoate 4, and 2,5-dimethyl-4-methoxy-3[2H]-furanone 5, each originating from the fruit, the delta(13)C(VPDB) and delta(2)H(VSMOW) data of commercial synthetic 1-5 and "natural" (biotechnologically derived) 1-4 were determined. With delta(13)C(VPDB) data of pineapple volatiles 1-4 varying from -12.8 to -24.4 per thousand, the range expected for CAM metabolism was observed. Compound 5 showed higher depletion from -20.9 to -28.6 per thousand. A similar situation was given for the delta(2)H(VSMOW) values of 3-5 from pineapple ranging from -118 to -191 per thousand, whereas 1 and 2 showed higher depleted values from -184 to -263 per thousand. In nearly all cases, analytical differentiation of 1-5 from pineapple and natural as well as synthetic origin was possible. In general, natural and synthetic 1-5 exhibited delta(13)C(VPDB) data ranging from -11.8 to -32.2 per thousand and -22.7 to -35.9 per thousand, respectively. Their delta(2)H(VSMOW) data were in the range from -242 to -323 per thousand and -49 to -163 per thousand, respectively.     

Use of a quantitative TaqMan-PCR for the fast quantification of mycobacteria in broth culture, eukaryotic cell culture and tissue

The quantification of slow-growing mycobacteria such as Mycobacterium tuberculosis or M. bovis from in vitro and in vivo samples is complicated by their long generation time, their ability to form aggregates, and their capacity to persist in a state of dormancy. We compared different methods for the establishment of growth curves for broth cultures of M. bovis bacille Calmette-Guérin (BCG). A quantitative TaqMan-PCR yielded results comparable with those obtained by protein quantification and measurement of the ATP content of the cultures. The quantitative TaqMan-PCR furthermore turned out to be particularly suitable for the measurement of multiplication of BCG within eukaryotic cells. Furthermore, it is a fast method allowing an estimation of the mycobacterial load in tissue long before colony counts can be obtained.     

The relationship of plasmids from environmental Yersinia isolates and the virulence plasmid of enteropathogenic Yersinia strains

The human pathogenic strains of Yersinia harbour a conserved plasmid carrying the Yop virulon. The virulence plasmid of Yersinia enterocolitica strains belonging to the serogroups O:3 and O:9 were used as probes to detect homologous sequences in plasmids of "avirulent" Yersinia strains. "Avirulent" Yersinia strains (Y. enterocolitica biogroup 1A, Y. intermedia, Y. kristensenii and Y. frederiksenii) lack the virulence plasmid. They are widely distributed in the environment and can frequently be isolated from clinical samples. Hybridisation experiments revealed a number of common genetic elements of the virulence plasmid and the plasmids of "avirulent" Yersinia strains. These elements were identified as genes involved in plasmid replication, as an endonuclease gene and as mobile genetic elements. However, none of the plasmid encoded virulence genes was present in the plasmids of "avirulent" Yersinia strains. The frequent occurrence and the possible etiological relevance of "avirulent" isolates will be discussed.     

Phosphorylation of muscle membranes: identification of a membrane-bound protein kinase

A membrane-bound protein kinase occurs in membranes derived from rat skeletal muscle and appears limited to a surface membrane fraction. The enzyme is magnesium dependent, is only minimally stimulated by cyclic nucleotides, and phosphorylates serine and to a lesser extent threonine residues of three membrane proteins with molecular weights of less than 30,000.     

Post-mortem findings in 54 cases of anesthetic associated death in cats from two spay-neuter programs in New York State

Anesthetic-associated death (AAD) in cats is infrequent, but occurs far more frequently than in people. Post-mortem investigations of AAD in cats are uncommon, and results only sporadically published. Here we report the findings in 54 cases of AAD in cats. Significant gross and/or microscopic pre-existing disease, including pulmonary, cardiac, and systemic disease, was detected in 33% of cases. Pulmonary disease was most frequently diagnosed (24% of cases), and included cases of Aelurostrongylus abstrusus infection (9% of cases). Heart disease, including two cases of hypertrophic cardiomyopathy, was less frequent (11% of cases). Four percent died from surgical complications. No?significant gross or microscopic disease was detected in 63% of cases. Additional studies are needed to determine if these findings are representative of AAD in cats in other geographic areas or with access to veterinary care. This study demonstrates that post-mortem investigation of AADs is an important and worthwhile endeavor.