Molecular characterization of non-aureus Staphylococcus spp. from heifer intramammary infections and body sites

The purpose of this study was to investigate non-aureus Staphylococcus spp. intramammary infections (IMI) in periparturient heifers and determine the relationship of precalving body site isolation with precalving IMI and postcalving IMI using molecular speciation and strain-typing methods. Primiparous heifers were enrolled at approximately 14 d before expected calving date. Precalving mammary quarter secretions and body site swabbing samples (teat skin, inguinal skin, muzzle, and perineum) were collected. Postcalving, mammary quarter milk samples were collected for culture and somatic cell counting. Precalving body site samples were cultured, and up to 10 staphylococcal colonies were saved for characterization. Staphylococcal isolates were speciated using matrix-assisted laser/desorption ionization time-of-flight mass spectrometry or sequencing of rpoB or tuf. Pulsed-field gel electrophoresis was used to strain type a subset of isolates. Overall, Staphylococcus chromogenes, Staphylococcus agnetis, and Staphylococcus simulans were the most common species identified in precalving mammary secretions, whereas S. chromogenes, Staphylococcus xylosus, and S. agnetis were the most common species found in postcalving milk samples. The most common species identified from body site samples were S. chromogenes, S. xylosus, and Staphylococcus haemolyticus. Mammary quarters that had a precalving mammary secretion that was culture positive for S. agnetis, S. chromogenes, or Staphylococcus devriesei had increased odds of having an IMI with the same species postcalving. A S. chromogenes IMI postcalving was associated with higher somatic cell count when compared with postcalving culture-negative quarters. Among heifers identified with a non-aureus Staphylococcus spp. IMI either precalving or postcalving, heifers that had S. agnetis or S. chromogenes isolated from their teat skin had increased odds of having the same species found in their precalving mammary secretions, and heifers with S. chromogenes, S. simulans, and S. xylosus isolated from their teat skin precalving were at increased odds of having an IMI with the same species postcalving. Overall, 44% of all heifers with a S. chromogenes IMI around the time of parturition had the same strain isolated from a body site. Based on pulsed-field gel electrophoresis, a high level of strain diversity was found.     

Strategic application of convective cooling to maximize the thermal gradient and reduce heat stress response in dairy cows

This study determined the effectiveness of convective cooling at different times of day when air temperature (T_a) was cycled from day to night. Mid-lactation Holstein cows (n = 12) were placed in 3 environmental chambers (4 cows per chamber) and acclimated to T_a 19.9°C (thermoneutral; TN) for 7 d followed by an incremental increase over 3 d to a heat stress (HS) condition. Conditions were maintained for 11 d at high and low daily T_a of 33 and 23°C, respectively. To determine adaptive HS response, the HS period was divided into early (E: d 11 to 14) and late (L: d 17 to 20) periods. During HS, cows were exposed to continuous fan (convective) cooling (CC), 8-h day fan cooling (1100 to 1900 h; DC), or 8-h night fan cooling (2300 to 0700 h; NC). Compared with DC, the NC treatment maximized the thermal gradient during the convective cooling. Each animal received all treatments within 3 trials using a repeated 3 × 3 Latin square design. Cows were fed a total mixed ration and milked twice daily. Thermal status was assessed by using thermal conductance and average daily values for mean, minimum, and maximum rectal temperature (T_re), skin temperatures, and respiration rate. Percent reduction in dry matter intake from TN to HS was less for CC than DC and NC, with no change from E to L periods. The DC group exhibited the greatest trend for a percent reduction in total milk yield below CC due to the significantly lower morning milk production. All values for total daily milk production decreased from E to L periods, with E to L reductions in both morning and afternoon milk production. Minimum T_re for CC and NC cows was 0.4°C below DC. In contrast, maximum T_re was similar for NC and DC groups, at 0.5 to 0.6°C above the CC group. Skin temperature for CC cows was always less than DC cows. Skin temperature for NC cows was equal to CC for minimum skin temperature, but exceeded both CC and DC cows for maximum skin temperature. Average skin temperature decreased from E to L, which suggested heat adaptation. The thermal advantage of night (lowest T_a and greatest thermal gradient) versus day cooling (greatest T_a and lowest thermal gradient) was increased heat transfer via thermal conductance with NC. The higher thermal strain of DC cows caused a larger percent decrease in morning milk yield than for NC cows. In contrast, use of convective cooling at night in the absence of elevated humidity could sufficiently reduce heat strain beyond DC to maintain milk production at a level that is closer to that of CC cows.     

Distribution of the alpha1 to alpha6 chains of type IV collagen in bovine follicles

During follicular development the proliferative and differentiated state of the epithelioid granulosa cells changes, and the movement of fluid across the follicular basal lamina enables the formation of an antrum. Type IV collagen is an important component of many basal laminae. Each molecule is composed of three alpha chains; however, six different type IV collagen chains have been identified. It is not known which of these chains are present in the follicular basal lamina and whether the type IV collagen composition of the basal lamina changes during follicular development. Therefore, we immunolocalized each of the six chains in bovine ovaries using antibodies directed to the nonconserved non-collagenous (NC) domains. Additionally, dissected follicles were digested with collagenase to release the NC domains, and the NC1 domains were then detected by standard Western immunoblot methods. The follicular basal lamina of almost all primordial and preantral follicles was positive for all type IV collagen alpha chains. Colocalization of type IV collagen and factor VIII-related antigen allowed for discrimination between the follicular and endothelial basal laminae. Type IV collagen alpha1, alpha2, alpha3, alpha4, and alpha5 chains were present within the follicular basal lamina of only a proportion of antral follicles (17 of 22, 20 of 21, 15 of 18, 14 of 28, and 12 of 23, respectively), and staining was less intense than in the preantral follicles. Staining for the alpha1 and alpha2 chains was diffusely distributed throughout the theca in regions not associated with recognized basal laminae. The specificity of this immunostaining for alpha1 and alpha2 chains of type IV collagen was confirmed by Western immunoblots. As well as being detected in the basal lamina of approximately half of the antral follicles examined, type IV collagen alpha4 also colocalized with 3beta-hydroxysteroid dehydrogenase-immunopositive cells in the theca interna. Type IV collagen alpha6 was detected in the basal lamina of only one of the 16 antral follicles examined. Thus, the follicular basal lamina changes in composition during follicular development, with immunostaining levels being reduced for all type IV collagen chains and immunoreactivity for type IV collagen alpha6 being lost as follicle size increases. Additionally, immunoreactivity for alpha1 and alpha2 appears in the extracellular matrix of the theca as it develops.     

Lazaroid improves intestinal blood flow in the rat during hyperdynamic bacteraemia

BACKGROUND: Intestinal mucosal hypoperfusion and loss of barrier function during sepsis may contribute to maintaining the septic state. Free radicals are produced during sepsis and antioxidants improve survival from experimental sepsis. It is unclear whether endothelial cell injury from free radicals results in altered microvascular reactivity. Lazaroids are antioxidants which scavenge radicals and block lipid radical chain reactions. The authors sought to determine whether lazaroids altered the intestinal microvascular responses to sepsis. METHODS: In vivo video microscopy was used to study the ileal microcirculation of the rat. A1 (inflow) arteriolar diameter and flow, A3 (premucosal) arteriolar diameters, and cardiac output were measured. Lazaroid or vehicle was infused before a bolus injection of live Escherichia coli or saline. RESULTS: Lazaroid alone had no effect on the intestinal vessels or haemodynamics. E. coli caused vasoconstriction (A1, -21 per cent, A3, -19 per cent of baseline) and hypoperfusion (-36 per cent) despite increased cardiac output (+31 per cent). Lazaroid significantly attenuated both constriction (A1, -11 per cent; A3, 10 to -1 per cent) and hypoperfusion (-15 per cent), but did not increase cardiac output (30 per cent). CONCLUSION: E. coli bacteraemia led to intestinal vasoconstriction and hypoperfusion. Lazaroid reduced this effect without altering central haemodynamic responses, suggesting that free radicals have a deleterious effect on the intestinal microcirculation during bacteraemia.     

Spark Plasma Sintering (SPS) of NASICON Ceramics

Spark plasma sintering (SPS) method was used to obtain dense NASICON ceramics with a high-electrical conductivity, which was compared with conventional solid-state sintering. The fully dense NASICON was achieved at a relatively low-sintering temperature of 1100°C, whereas the apparent density of the specimen prepared by conventional sintering was 74% of the theoretical density. The highest conductivity of 1.8 × 10⁻³ Scm⁻¹ at 25°C, which is comparable to the best value reported, was achieved using the SPS process. Considering the phase, density, and microstructure, it appears that there is more room for improved conductivity by controlling the amount of monoclinic zirconia and the resistive grain-boundary glass phase.     

Profiling care and benchmarking best practice in care of hospitalized elderly: the Geriatric Institutional Assessment Profile

This article reports on a new instrument, the Geriatric Institutional Assessment Profile (GIAP), developed to assess (1) hospital workers' knowledge, attitudes, and perceptions regarding care of geriatric patients, and (2) the perceived adequacy of an institutional environment to serve geriatric patients' needs. Findings are reported from 303 questionnaires completed by health care employees from a 658-bed academic medical center. Internal consistency estimates were consistently high for the various components of the GIAP. Factor analysis was performed to examine underlying dimensions of knowledge and institutional environment. The GIAP has the potential to narrow the gap between actual and best practice in geriatric care by identifying staff information needs and concerns, as well as institutional barriers and facilitators to providing quality geriatric hospital care.     

Soil column evaluation of factors controlling biodegradation of DNT in the vadose zone

High concentrations of 2,4-dinitrotoluene (2,4-DNT) and 2,6-dinitrotoluene (2,6-DNT) are present in vadose zone soils at many facilities where explosives manufacturing has taken place. Both DNT isomers can be biodegraded under aerobic conditions, but rates of intrinsic biodegradation observed in vadose zone soils are not appreciable. Studies presented herein demonstrate that nutrient limitations control the onset of rapid 2,4-DNT biodegradation in such soils. In column studies conducted at field capacity, high levels of 2,4-DNT biodegradation were rapidly stimulated by the addition of a complete mineral medium but not by bicarbonate-buffered distilled deionized water or by phosphate-amended tap water. Biodegradation of 2,6-DNT was not observed under any conditions. Microcosm studies using a DNT-degrading culture from column effluent suggest that, after the onset of 2,4-DNT degradation, nitrite evolution will eventually control the extent of degradation achieved by two mechanisms. First, high levels of nitrite (40 mM) were found to strongly inhibit 2,4-DNT degradation. Second, nitrite production reduces the solution pH, and at pH levels below 6.0, 2,4-DNT degradation slows rapidly. Under conditions evaluated in laboratory-scale studies, 2,4-DNT biodegradation enhanced the rate of contaminant loss from the vadose zone by a factor of 10 when compared to the washout due to leaching.