Proliferation of peripheral blood mononuclear cells from healthy piglets after mitogen stimulation as indicators of disease resilience     

Ryan L Jeon  Caroline Gilbert  Jian Cheng  Austin M Putz  Mike K Dyck  Graham S Plastow  Frederic Fortin  PigGen Canada  Jack C M Dekkers  John C S Harding 《Journal of animal science》2021,99(8)

Disease resilience refers to the productivity of an animal under disease. Given the high biosecurity of pig nucleus herds, traits that can be measured on healthy pigs and that are genetically correlated with disease resilience, that is, genetic indicator traits, offer a strategy to select for disease resilience. Our objective was to evaluate mitogen stimulation assays (MSAs) on peripheral blood mononuclear cells (PBMCs) from young healthy pigs as genetic indicators for disease resilience. Data were from a natural disease challenge in which batches of 60 or 75 naïve Yorkshire × Landrace piglets were introduced every 3 wk into a continuous flow barn that was seeded with multiple diseases. In this environment, disease resilience traits, including growth, treatment, and mortality rates, were recorded on 3,136 pigs that were genotyped with a high-density marker panel. PBMCs from 882 of these pigs from 19 batches were isolated from whole blood collected prior to the disease challenge and stimulated with five mitogens: concanavalin A (ConA), phytohemagglutinin (PHA), pokeweed mitogen (PWM), lipopolysaccharide (LPS), and phorbol myristate acetate (PMA). The proliferation of cells was evaluated at 48, 72, and 96 h and compared with unstimulated samples (rest count). Heritabilities of cell proliferation were estimated using a model with batch as a fixed effect and covariates of entry age; rest count; complete blood count proportions of lymphocytes, monocytes, eosinophils, and basophils; and pen, litter, and animal genetics as random effects. Heritability estimates were highest for response to ConA (0.30 ± 0.09, 0.28 ± 0.10, 0.17 ± 0.10, and 0.25 ±0.10 at 48, 72, and 96 h after stimulation and for area under the curve across the three time points, respectively). Estimates were in a similar range for response to PHA and PMA but low for PWM and LPS. Responses to ConA, PHA, and PMA were moderately genetically correlated with several disease resilience traits and in the expected direction, but individual estimates were not significantly different from zero due to large SEs. In conclusion, although validation is needed, MSAss, in particular based on ConA, show promise as genetic indicator traits for disease resilience.  相似文献   

Production of a baculovirus-derived gp50 protein and utilization in a competitive enzyme-linked immunosorbent assay for the serodiagnosis of pseudorabies virus.   总被引：2，自引：0，他引：2       下载免费PDF全文

I Prud''homme  E M Zhou  M Traykova  H Trotter  M Chan  A Afshar    M J Harding 《Canadian journal of veterinary research》1997,61(4):286-291

The pseudorabies virus (PRV) gp50 envelope glycoprotein gene was cloned and expressed in a recombinant baculovirus. An anti-gp50 Mab (1842) recognized a protein of approximately 40 kDa in immunoblotting assays from infected insect cell lysates, while this product was not present in cells infected with wild-type baculovirus. The recombinant protein was purified by lectin affinity chromatography, utilizing lectins specific for O-linked oligosaccharides (Artocarpus integrifolia and Glycine max). Competitive (c) ELISAs, using either crude or lectin-purified antigen, were devised for the detection of antibodies to PRV in sera, and were capable of monitoring sero-conversion by day 14 post-infection. Furthermore, a specificity of 100% and sensitivity of 98% (crude lysate antigen) or 96% (lectin-purified antigen) was found for a panel of 80 swine sera, using the cELISA, as compared to a serum neutralization (SN) test. These studies demonstrated that recombinant PRV gp50 protein shows promise as a cELISA antigen, for serodetection of PRV.  相似文献   

Effect of Cryopreservation on Bovine Sperm Viability as Determined by Dual DNA Staining     

DL Garner  CA Thomas  CH Allen  PL Senger  RG Sasser 《Reproduction in domestic animals》1997,32(6):279-283

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Specificity and nucleotyping studies of a gp50-based polymerase chain reaction assay for detection of pseudorabies virus.   总被引：7，自引：1，他引：6  

M J Harding  I Prud''homme    J Rola 《Canadian journal of veterinary research》1997,61(2):157-160

PCR primers that amplify a region of the gp50 envelope glycoprotein gene of a number of vaccinal and field strains of pseudorabies virus (PRV) have been previously described (Galeota-Wheeler and Osorio, Am J Vet Res 1991: 52; 1799-1803). This gp50-based PCR assay was tested for its diagnostic applicability, utilizing a panel of nine PRV isolates and 13 related herpesviruses, originating from domestic animal species and man. Slight modifications to the original PRV PCR protocol ensured that false positive PCR products from avian herpesviruses were not evident in agarose gel electrophoresis analysis. Nucleotide sequence data derived from the PCR product revealed that the region of the genome amplified was markedly conserved and allowed only for virus subgrouping, rather than definitive isolate characterization.  相似文献   

Effect of the environment on seed yield in bulk populations of lima beans     

C. L. Tucker  James Harding 《Euphytica》1974,23(1):135-139

Summary Two bulk populations of lima beans were developed from biparental crosses of cultivars. Each bulk population was grown in two different sites in the same commercial growing area in California for 9 generations. They were yield-tested in one of the two environments, designated test location. The populations grown in the test location had higher yields than the population grown in the non-test location and showed significant divergence in yield from the F₉ to F₁₁ generations. Yields of the bulk populations were intermediate to parental yields.  相似文献   

Samples sizes required to accurately quantify viral load and histologic lesion severity at the maternal–fetal interface of PRRSV-inoculated pregnant gilts     

Carolina M. Malgarin  Javier B. Zarate  Predrag Novakovic  Susan E. Detmer  Daniel J. MacPhee  John C. S. Harding 《Journal of veterinary diagnostic investigation》2021,33(2):322

Porcine reproductive and respiratory syndrome virus (PRRSV) is transmitted vertically, causing fetal death in late gestation. Spatiotemporal distribution of virus at the maternal–fetal interface (MFI) is variable, and accurate assessment of viral concentration and lesions is thus subject to sampling error. Our objectives were: 1) to assess whether viral load and lesion severity in a single sample of endometrium (END) and placenta (PLC), collected near the base of the umbilical cord (the current standard), are representative of the entire organ; and 2) to compare sampling strategies and evaluate if spatial variation in viral load can be overcome by pooling of like-tissues. Spatially distinct pieces of END and PLC of 24 fetuses from PRRSV-2–infected dams were collected. PRRSV RNA quantified by RT-qPCR was compared in 5 individual pieces per fetus and in respective pools of tissue and extracted RNA. Three distinct pieces of MFI were assessed for histologic severity. Concordance correlation and kappa inter-rater agreement were used to characterize agreement among individual samples and pools. The viral load of individual samples and pools of END had greater concordance to a referent standard than did samples of PLC. Larger pool sizes had greater concordance than smaller pool sizes. Average viral load and lesion severity did not differ by location sampled, and no technical advantages of pooling tissues versus RNA extracts were found. We conclude that multiple pieces of MFI tissues must be evaluated to accurately assess lesion severity and viral load. Three pieces per fetus provided a reasonable balance of cost and logistic feasibility.  相似文献   

高大冷杉温室Strophosoma象甲的微生物防治     

夏洛蒂· 尼尔森  Jφrgen Eilenberg  Jφrgen Eilenberg 《安徽农业大学学报》2007,34(2):185-194

In Denmark, the weevils Strophosoma melanogrammum and S.capitatum cause economic damage in Noble fir due to the adult stage feeding on the needles.No chemical treatments of these weevils are allowed in Denmark,so biological control is an attractive solution.We evaluated the potential for microbial control of larvae of Strophosoma spp.based on laboratory bioassays and field applications,taking effect on both target and non-target into consideration,as well as persistence of the applied fungus.In the laboratory Beauveria bassiana,Paecilomyces farinosus and Metarhizium anisopliae were able to infect and cause mycosis in Strophosoma larvae.Among the tested isolates the most virulent isolate was M.anisopliae BIPESCO 5,which resulted in 80 % mortality.In the field experiment M.anisopliae,isolate BIPESCO 5,was applied to the soil as a conidial suspension against larvae of Strophosoma spp.The effect of the fungus on the target population was monitored at a weekly basis by counts of emerging adult weevils during their activity periods.The population of Strophosoma spp.was reduced by up to 60% in treated plots compared to control plots.The non-target effects of M.anisopliae were studied by sampling insects and ticks from both treated and control plots.Seven days after treatment,two sampled insect orders (Hemiptera and Coleoptera) and ticks were found with prevalences of M.anisopliae above 50%,compared to no infection in the insects collected from control plots.Infections in coccinellids were found as long as 277 days after treatment.However,the effect on population level of non-target is still unexplored.The persistence of the fungus was documented by plating a soil suspension onto agar.We documented that conidia of M.anisopliae could persist in the greenery plantation for at least 418 days after application.  相似文献   

Cyclophilin: a specific cytosolic binding protein for cyclosporin A   总被引：89，自引：0，他引：89  

R E Handschumacher  M W Harding  J Rice  R J Drugge  D W Speicher 《Science (New York, N.Y.)》1984,226(4674):544-547

Cyclophilin, a specific cytosolic binding protein responsible for the concentration of the immunosuppressant cyclosporin A by lymphoid cells, was purified to homogeneity from bovine thymocytes. Cation-exchange high-performance liquid chromatography resolved a major and minor cyclophilin species that bind cyclosporin A with a dissociation constant of about 2 X 10(-7) moles per liter and specific activities of 77 and 67 micrograms per milligram of protein, respectively. Both cyclophilin species have an apparent molecular weight of 15,000, an isoelectric point of 9.6, and nearly identical amino acid compositions. A portion of the NH2-terminal amino acid sequence of the major species was determined. The cyclosporin A-binding activity of cyclophilin is sulfhydryl dependent, unstable at 56 degrees C and at pH 4 or 9.5, and sensitive to trypsin but not to chymotrypsin digestion. Cyclophilin specifically binds a series of cyclosporin analogs in proportion to their activity in a mixed lymphocyte reaction. Isolation of cyclophilin from the cytosol of thymocytes suggests that the immunosuppressive activity of cyclosporin A is mediated by an intracellular mechanism, not by a membrane-associated mechanism.  相似文献   

Sex attractant of sugar beet wireworm: identification and biological activity   总被引：2，自引：0，他引：2  

M Jacobson  C E Lilly  C Harding 《Science (New York, N.Y.)》1968,159(811):208-210

The sex attractant produced by adult females of the sugar beet wire-worm, Limonius californicus (Mannerheim) has been isolated and identified as valeric acid. In the laboratory, male wireworm beetles are repelled by the pure attractant but are drawn with intense sexual excitement to its dilute solutions; in the field, male beetles are lured from a distance of 12 meters. The pheromone occurs in unusually large amounts in the female's body.  相似文献   

THE BORON CONTENT OF SEA WATER     

Moberg EG  Harding MW 《Science (New York, N.Y.)》1933,77(2004):510-511

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