Function of neutrophils and chemoattractant properties of fetal placental tissue during the last month of pregnancy in cows.

Neutrophils were isolated from the blood of pregnant cows on days 255, 265, and 275 of pregnancy, and on the day of parturition (n = 5/group), and in addition, simultaneously from 4 ovariectomized healthy cows (control animals). Neutrophils were subjected to neutrophil function assays (chemotaxis against zymosan-activated serum, random migration, ingestion of 125I-iododeoxyuridine [IdUR]-labeled Staphylococcus aureus, iodination of proteins, cytochrome C reduction, antibody-independent and antibody-dependent cell-mediated cytotoxicity). Results were expressed as percentage of control animals. Fetal placental tissue (cotyledon), uterine wall tissue, and skeletal muscle were obtained from the principal animals on the aforementioned days via laparotomy, and tissue suspensions were prepared. Chemotaxis of neutrophils was tested against tissue supernatants. Compared with day 255, there was an increase in ingestion of 125I-IdUR-S aureus at parturition, whereas iodination of proteins and cytochrome C reduction were reduced on the day of calving. The other neutrophil functions tested did not change over time of gestation. Fetal placental and uterine wall tissue attracted neutrophils with uterine wall tissue having a tendency to be more potent than cotyledonary tissue. Skeletal muscle tissue did not attract neutrophils. There was no change in chemotaxis response of neutrophils evoked by intrauterine and uterine tissues over time of gestation. It was concluded that at parturition, neutrophil function is impaired with respect to their bactericidal effects, which may render the animal more susceptible to bacterial infections, and that the chemoattractant properties of fetal placental and uterine wall tissues are tissue-specific, at least when compared with skeletal muscle.(ABSTRACT TRUNCATED AT 250 WORDS)     

Depression of polymorphonuclear leukocyte function associated with experimentally induced Escherichia coli mastitis in sows

In a study of susceptibilities of sows from 2 herds to experimentally induced Escherichia coli mastitis, a marked difference was seen. The "susceptible" sows were from a conventional herd and "resistant" sows were from a specific-pathogen-free herd. The purpose of the study was to determine whether deficient neutrophil function was associated with increased susceptibility to E coli-induced mastitis. Four in vitro procedures were used to evaluate polymorphonuclear leukocyte (PMN) function: (i) random migration under agarose, (ii) ingestion of 125I-iododeoxyuridine-labeled Staphylococcus aureus, (iii) quantitative nitroblue tetrazolium reduction, and (iv) iodination. After parturition and intramammary inoculation with E coli, sows from the susceptible herd were neutropenic and the neutrophils which were present in the peripheral blood had reduced function. Specifically, there were depressed random migration under agarose, S aureus ingestion, and iodination when compared with PMN function in resistant sows. These data indicate that susceptibility to E coli mastitis was associated with deficiencies in PMN numbers and function. Potential causes of the neutrophil dysfunction are discussed and include possible systemic hormonal aberrations or the presence of an inapparent viral or bacterial infection.     

Identification of ALS inhibitor‐resistant Amaranthus biotypes using polymerase chain reaction amplification of specific alleles

Summary Polymerase chain reaction (PCR) amplification of specific alleles (PASA) was adapted as a molecular marker‐based method for the rapid detection of point mutations in Amaranthus retroflexus and Amaranthus rudis leading to ALS inhibitor resistance. Two pairs of primers were designed for the specific amplification of alleles of the ALS gene of susceptible and resistant biotypes. The allele‐specific primer matched the desired allele, but mismatched the different allele at its 3′ end. Differentiation was carried out by comparison of the amplified DNA fragments in gel electrophoresis after PASA‐PCR. In A. rudis, differentiation was possible with one PCR and genomic DNA as probe. A ‘nested’ PCR was necessary for the differentiation of sensitive and resistant A. retroflexus. PASA is useful for the identification of resistant weed biotypes and also as a monitoring tool to map resistance occurrence and distribution. Advantages include the fast and clear separation of those plants with and without mutations at an early stage of development, its easy and consistent performance and quick results compared with existing resistance detection tests. These advantages, when combined with management strategies, enable further activities to reduce herbicide resistance.     

Root architecture of sorghum genotypes differing in root angles under different water regimes

Plant root architecture offers the potential for increasing soil water accessibility, particularly under water-limited conditions. The objectives of this study were to evaluate the root architecture in two genotypes of sorghum (Sorghum bicolor (L.) Moench) differing in root angles and to assess the influence of different deficit irrigation regimes on root architecture. The response of two sorghum genotypes, ‘Early Hegari-Sart’ (EH; steep root angle) and ‘Bk7’ (shallow root angle) to four irrigation treatments was investigated in two replicated outdoor studies using large pots. The results indicated that EH possessed steeper brace and crown root angles, fewer brace roots, greater root biomass, and root length density than Bk7 at deeper soil depths (i.e., 15–30 and 30–45 cm) compared with a shallower depth (i.e., 0–15 cm). Across the soil profile, EH had greater root length density and length of roots of small diameter (<1 mm) than Bk7. Accordingly, EH showed more rapid soil-water capture than Bk7. Different levels of irrigation input greatly affected root architecture. Severe deficit irrigation (25% of full crop transpiration throughout the season) increased the angle and number of crown roots, root biomass, and root length density compared with 75 and 100% of full crop transpiration treatments. Consequently, root system architecture can be effectively manipulated through both genotypic selection and irrigation management to ensure optimal performance under different levels of soil available water.     

Validating the effectiveness of alternative euthanasia techniques using penetrating captive bolt guns in mature swine (Sus scrofa domesticus)

Euthanasia of mature swine is challenging. Temporal and behind-the-ear locations are two sites that have been identified as alternatives to the more commonly used frontal placement. In stage one, the effectiveness of two penetrating captive bolt gun styles (cylinder or pistol) was evaluated using frontal, temporal, and behind-the-ear placement in anesthetized mature swine (n = 36; weight: 267 ± 41 kg). For stage one, when evaluating treatment efficacy by sex, the cylinder-style equipment was 100% effective in achieving death when applied to all cranial locations (frontal, temporal, and behind-the-ear) for sows; however, the pistol-style equipment was only 100% effective when applied at the behind-the-ear location for sows. For boars, the cylinder-style equipment was 100% effective when applied to the frontal and behind-the-ear location, but the pistol-style equipment was not effective for any cranial location in boars. Therefore, the pistol–frontal, pistol–temporal, pistol–behind-the-ear, and cylinder–temporal were not included for boars, and pistol–frontal and pistol–temporal were not included for sows in stage two. In stage two, commercial, mixed-breed, mature swine (n = 42; weight: 292 +/− 56 kg) were randomly assigned to one of four treatments based on the inclusion criteria described in stage one. A three-point traumatic brain injury (TBI) score (0 = normal; 1 = some abnormalities; 2 = grossly abnormal, unrecognizable) was used to evaluate six neuroanatomical structures (cerebral cortex, cerebellum, hypothalamus, thalamus, pons, and brain stem), and the presence of hemorrhage was also noted. All treatments were 100% effective in stage two. A significant interaction between gun style and placement was determined on predicting total TBI as the cylinder style produced a higher total TBI score compared with the pistol type of the magnitude of +2.8 (P < 0.01). The cylinder style tended to produce a greater TBI score than the pistol in the temporal location (+1.2; P = 0.08). No difference was noted for TBI score behind-the-ear between the cylinder- and pistol-style gun (P > 0.05). TBI tended to be less in boars compared with sows (−0.6; P = 0.08). Hemorrhage was observed in frontal, parietal, occipital, and temporal lobes. This study demonstrated that the cylinder-style captive bolt gun more effectively resulted in brain trauma and death compared with a pistol-style gun and the behind-the-ear and temporal placement showed promise as an alternative placement site for euthanizing mature pigs on-farm.     

Free-range system and supplementation of 25-hydroxicholecalciferol increases the performance and serum vitamin levels in mixed-parity sows

Improvements in sow productivity have raised questions regarding dietary vitamin D recommendations. The present study aimed to evaluate the effects of the housing system with access to sunlight exposure and supplementation of 25-hydroxicholecalciferol on performance and serum levels of 25(OH)D₃ in sows during gestation and lactation. Sows were distributed in an experimental design with two housing systems: gestation crates or gestation free-range system with external area for sunlight exposure; and two diets: 0 or 50 μg of 25-hydroxicholecalciferol kg⁻¹. The use of 25-hydroxicholecalciferol tended (P = 0.052) to improve total born and influenced (P = 0.046) on number of born alive. Litter weight at birth was also increased (P = 0.01) by 25-hydroxicholecalciferol supplementation; 25-hydroxicholecalciferol supplementation and housing system (free-range with sunlight exposure) tended to increase weaning weight (P = 0.07) and litter daily gain (P = 0.051) during lactation. Exposure to sunlight and 25-hydroxicholecalciferol supplementation increased 25(OH)D₃ serum levels when compared with control treatment during gestation (136.95 vs. 113.92 ng mL⁻¹; P = 0.035) and lactation (120.29 vs. 88.93 ng mL⁻¹; P = 0.026). In conclusion, the association of 25-hydroxicholecalciferol supplementation with exposure to sunlight during gestation improved significantly 25(OH)D₃ serum levels and consequently performance traits in gestation and lactation.     

Cholera toxin promotes the generation of semi-mature porcine monocyte-derived dendritic cells that are unable to stimulate T cells

Cholera toxin (Ctx) is a powerful mucosal adjuvant with potential applications for oral vaccination of swine. Dendritic cells (DC) play a key role in the decision between immunity and tolerance, and are likely target cells for mediating Ctx functions in vivo. Therefore, we examined the capacity of Ctx to enhance stimulatory activity of porcine monocyte-derived DC (MoDC). Ctx promoted the development of a semi-mature DC phenotype, with decreased levels of MHC class II and CD40, but increased CD80/86 expression. These changes were associated with activation of extracellular signal-regulated kinase (ERK), but not NFkappaB or c-Jun N-terminal kinase (JNK). Functionally, Ctx-priming greatly diminished T cell stimulatory capacity both in antigen-specific and superantigen-induced proliferation assays. The lower proliferation rate was not due to increased apoptosis of either DC or T cells. Ctx suppressed TNFalpha secretion by MoDC, but induced IL-10 production. The observed effects on T cell proliferation could only be partially mimicked by IL-10 alone. However, addition of recombinant TNFalpha to co-cultures of Ctx-primed MoDC and lymphocytes restored lymphocyte proliferation in a concentration-dependent manner. Ctx-primed DC were not actively tolerogenic, since they could not suppress proliferative T cell reactions induced by untreated DC.     

Variance estimation for population attributable risk in a complex cross-sectional animal health survey

Population attributable risk estimates offer a method of combining information on population exposure and disease risk factors into a single measure. Univariate and multivariable methods exist for calculating point estimates and variances under the assumption of equal sampling probabilities. National Animal Health Monitoring System national studies typically use a complex survey design (where selection probabilities vary by design strata), which makes use of these methods of calculating variance inappropriate. We suggest the use of a method called "delete-a-group" jackknife to estimate the variance of population attributable risk when a complex survey design has been implemented. We demonstrate the method using an example of Johne's disease. Advantages of the "delete-a-group" jackknife method include simplicity of implementation and flexibility to estimate variance for any point estimate of interest.