Cytokine response of porcine cell lines to Salmonella enterica serovar typhimurium and its hilA and ssrA mutants

Salmonella enterica serovar Typhimurium (S. Typhimurium) is a facultative intracellular bacterium which can infect and colonize pigs. After contact with enterocytes and macrophages, S. Typhimurium induces production of cytokines thus triggering the innate immune response. In this study we evaluated the cytokine response of two porcine cell lines, IPI-2I and 3D4/31, of epithelial or macrophage origins, respectively, to the wild-type S. Typhimurium and its hilA and ssrA mutants. We observed that the 3D4/31 cell line essentially did not respond to S. Typhimurium infection when a medium with foetal calf serum was used. However when the 3D4 cell line was incubated overnight in the presence of porcine serum, it efficiently responded to the wild-type strain and the ssrA mutant but not to the noninvasive hilA mutant as measured by mRNA quantification of TNF-alpha, IL-8 and GM-CSF by the real-time RT-PCR. In IPI-2I, all the cytokines were also induced by the wild-type S. Typhimurium and the ssrA mutant although the induction of TNF-alpha was lower than that induced by the wild-type strain. The hilA mutant was unable to induce any of the cytokines tested. The ssrA mutant can therefore be considered as more suitable for further vaccine development as the stimulation of innate immune response is important for animal protection against a challenge with virulent strains.     

Identification of key performance indicators for on-farm animal welfare incidents: possible tools for early warning and prevention

Background

The objective of this study was to describe aspects of case study herds investigated by the Department of Agriculture, Fisheries and Food (DAFF) in which animal welfare incidents occurred and to identify key performance indicators (KPIs) that can be monitored to enhance the Early Warning System (EWS). Despite an EWS being in place for a number of years, animal welfare incidents continue to occur. Questionnaires regarding welfare incidents were sent to Superintending Veterinary Inspectors (SVIs), resulting in 18 herds being chosen as case study herds, 12 of which had a clearly defined welfare incident date. For each study herd, data on six potential KPIs were extracted from DAFF databases. The KPIs for those herds with a clearly defined welfare incident date were studied for a consecutive four year window, with the fourth year being the ''incident year'', when the welfare incident was disclosed. For study herds without a clearly defined welfare incident date, the KPIs were determined on a yearly basis between 2001 and 2009.

Results

We found that the late registration of calves, the use of on-farm burial as a method of carcase disposal, an increasing number of moves to knackeries over time and records of animals moved to ''herd unknown'' were notable on the case farms.

Conclusion

Four KPIs were prominent on the case study farms and warrant further investigation in control herds to determine their potential to provide a framework for refining current systems of early warning and prevention.     

Observations on the cooling and cryopreservation of pig oocytes at the germinal vesicle stage

This study examined the viability of pig oocytes at the germinal vesicle stage following cooling or cryopreservation. Cumulus-intact oocytes (n = 641) were collected from slaughterhouse pig ovaries and used in two experiments. In Exp. I the viability of 1) control, 2) cryoprotectant control (CC, 1.5 M glycerol/.5 M sucrose), 3) cooled (0 degrees C) and 4) cryopreserved (-196 degrees C) oocytes was assessed after no incubation or a 24-h incubation. Survivability was judged by morphological appearance, trypan blue exclusion and fluorescein diacetate staining. Survival rate of control oocytes (90%; based primarily on morphological appearance of the cumulus) incubated 0 h was greater (P less than .05) than that of all other groups, whereas survival rate of -196 degrees C oocytes (57%) was less (P less than .05) than that of all other groups. However, vital staining of 0 degrees C and -196 degrees C oocytes showed 0% survival rate as evidenced by trypan blue uptake and lack of fluorescence. The cumulus cells surrounding oocytes that were stored at 0 degrees C or -196 degrees C survived freezing as evidenced by trypan blue exclusion and intense fluorescence. Similar differences among treatment groups were found for oocytes incubated 24 h. Exp. 2 examined the temperature at which oocytes became sensitive to cooling. Oocyte death occurred when oocytes were cooled to 15 degrees C or lower. These results demonstrate that pig oocytes at the germinal vesicle stage did not survive cooling to 15 degrees C or below. When assessing the viability of cryopreserved cumulus enclosed oocytes it is important to use vital stains in conjunction with morphological appearance.     

Prognosis for Neonatal Foals in an Intensive Care Unit

This study was conducted to develop an equation for the prediction of outcome in neonatal foals undergoing treatment in an intensive care unit (ICU). Fifty-three physical examination, historical, and clinicopathologic variables were analyzed from the records of 99 neonatal foals (<14 days of age) treated in the neonatal ICU of the Equine Medical Center. The outcome was recorded and the results were categorized into either surviving or nonsurviving groups. The mean values for the 2 groups were compared, and variables that differed significantly between the two groups were retained and used to construct a logistic regression equation. Retained variables were heart rate, temperature, and neutron phil count. The predictive equation then was tested prospectively in 2 additional groups of foals from 2 separate ICUs. The predicted outcome was compared to the actual outcome, and performance variables were calculated. Sensitivity (.83), specificity (.87), negative predictive value (.72), and positive predictive value (.93) were determined for foals from one neonatal ICU; the sensitivity (.83), specificity (.44), negative predictive value (.44), and positive predictive value (.83) were lower for foals at a second, separate ICU.     

Effects of intensive harvesting on nutrient capitals of three forest types in New England

Effects of whole-tree clearcutting are being studied in three major forest types in the northeastern United States: a spruce-fir forest in central Maine, a northern hardwood forest in New Hampshire, and a central hardwood forest in Connecticut. At each site we sampled total and extractable nutrient capitals, inputs and outputs of nutrient ions in precipitation and streamflow, nutrient removals in harvested products, and nutrient accumulation in regrowth. Depending upon location, combined losses of nutrients in harvested products and increased leaching to streams were in the ranges of 374–558 kg ha⁻¹ for Ca, 135–253 kg ha⁻¹ for K, 50–65 kg ha⁻¹ for Mg, 248–379 kg ha⁻¹ for N, and 19–54 kg ha⁻¹ for P. Opportunities for replacing these losses over the next rotation are best for N. Data on inputs in precipitation versus outputs in streamflow indicate that, once effects of harvest subside, most N in precipitation will stay within the forest. By contrast, Ca shows a net output of 8–15 kg ha⁻¹ year⁻¹ from uncut watersheds, and the added leaching losses due to harvest may have a serious impact on Ca capital. This is especially the case for the Connecticut site, where total site capital for Ca is only about 4000 kg ha⁻¹.     

Effects of Aspergillus oryzae fermentation extract on fermentation of amino acids, bermudagrass and starch by mixed ruminal microorganisms in vitro

The objective of this study was to examine the effects of Aspergillus oryzae fermentation extract (Amaferm) on the in vitro ruminal fermentation of coastal bermudagrass, soluble starch and amino acids. Mixed ruminal microorganisms were incubated in anaerobic media for either 24 h (Amaferm alone, soluble starch, amino acids) or 48 h (bermudagrass). Amaferm was added to the incubation bottles (n = 4) at concentrations of 0, .4 or 1.0 g/liter. When mixed ruminal microorganisms were incubated with only Amaferm, the 1.0 g/liter concentration increased the production of hydrogen (H2; P less than .001), methane (CH4; P less than .01), acetate (P less than .05), butyrate (P less than .01), total VFA (P less than .05) and NH3 (P less than .05). Addition of both levels of Amaferm to soluble-starch fermentations tended to enhance the production of H2 (P less than .11), CH4 (P less than .15), acetate (P less than .29) and total VFA (P less than .19); propionate production was increased (P less than .10) by 1.0 g/liter Amaferm, resulting in a decrease (P less than .05) in the acetate:propionate ratio. Fermentation of amino acids plus 1.0 g/liter Amaferm enhanced the production of acetate (P less than .05), propionate (P less than .05), valerate (P less than .01) and total VFA (P less than .10) and decreased the acetate:propionate ratio (P less than .05). In addition, NH3 production tended (P less than .19) to increase with both levels of Amaferm. When bermudagrass was the substrate, few changes in fermentation products were observed with Amaferm treatment.(ABSTRACT TRUNCATED AT 250 WORDS)     

Methane emissions and δ13C composition from beef steers consuming increasing proportions of sericea lespedeza hay on bermudagrass hay diets

An experiment was conducted to evaluate the effects of different proportions of ‘Au Grazer’ sericea lespedeza [SL, Lespedeza cuneata (Dum. Cours.) G. Don], a legume rich in condensed tannins (CT), on nutrient intake and digestibility, and to estimate methane (CH₄) emissions and ¹³C isotopic composition (δ¹³C_CH4) from beef steers consuming a forage-based diet. Twenty-five Angus-crossbred steers were distributed in a randomized complete block design (344 ± 48 kg initial BW), and randomly assigned to one of five treatments: 0SL, 25SL, 50SL, 75SL, and 100SL, diets containing 0%, 25%, 50%, 75%, and 100% of SL hay, respectively, mixed with ‘Tifton-85’ bermudagrass hay (Cynodon spp.). The study was carried out for two experimental periods of 21-d each. The statistical model included the fixed effect of treatment and random effects of block, experimental period, and their interaction. Apparent total tract digestibility of crude protein, neutral detergent fiber, and acid detergent fiber was linearly decreased (P < 0.001) by the inclusion of SL. No effects were observed for total CH₄ emissions per day, nor for CH₄ relative to organic matter intake or digestible organic matter with the inclusion of SL. However, emission of CH₄ in relation to intake of CT was affected by treatment (P < 0.001). A linear (P < 0.001) decrease and a quadratic effect (P < 0.001) were observed for δ¹³C of diets and gas, respectively, in which diets and enteric CH₄ with greater inclusion of SL were more depleted in ¹³C. Moreover, the difference in δ¹³C between diets and gas (Δδ¹³C) had a linear decrease (P = 0.001) with the inclusion of SL. The model developed to predict the C₃ proportions in the enteric CH₄ fitted to predicted values (P < 0.0001). Therefore, greater proportions of SL resulted in lesser CH₄ emission when CT intake was considered and the isotopic composition from enteric CH₄ was able to predict the contribution of SL in the emissions.