Water temperature affects the biotransformation and accumulation of a psychoactive pharmaceutical and its metabolite in aquatic organisms.

Pharmaceutically active compounds (PhACs) have been shown to accumulate in aquatic and riparian food-webs. Yet, our understanding of how temperature, a key environmental factor in nature, affects uptake, biotransformation, and the subsequent accumulation of PhACs in aquatic organisms is limited. In this study, we tested to what extent bioconcentration of an anxiolytic drugs (temazepam and oxazepam) is affected by two temperature regimes (10 and 20 °C) and how the temperature affects the temazepam biotransformation and subsequent accumulation of its metabolite (oxazepam) in aquatic organisms. We used European perch (Perca fluviatilis) and dragonfly larvae (Sympetrum sp.), which represent predator and prey species of high ecological relevance in food chains of boreal and temperate aquatic ecosystems. Experimental organisms were exposed to target pharmaceuticals at a range of concentrations (0.2-6 µg L-1) to study concentration dependent differences in bioconcentration and biotransformation. We found that the bioconcentration of temazepam in perch was significantly reduced at higher temperatures. Also, temperature had a strong effect on temazepam biotransformation in the fish, with the production and subsequent accumulation of its metabolite (oxazepam) being two-fold higher at 20 °C compared to 10 °C. In contrast, we found no temperature dependency for temazepam bioconcentration in dragonfly larvae and no detectable biotransformation of the parent compound that would result in measurable concentrations of oxazepam in this organism. Our results highlight that while organisms may share the same aquatic ecosystem, their exposure to PhACs may change differently across temperature gradients in the environment.

Voluntary intake and digestibility by sheep of alfalfa ensiled at different moisture concentrations following fertilization with dairy slurry.

Dairy slurry is used commonly as an animal-sourced fertilizer in agronomic production. However, residual effects of slurry application on intake and digestibility of alfalfa (Medicago sativa L.) silage from subsequent harvests are not well known. The objective of this study was to determine if moisture concentration of alfalfa silage and timing of dairy slurry application relative to subsequent harvest affected intake and digestibility by sheep. Katahdin crossbred ewes (n = 18; 48 ± 5.3 kg) in mid-gestation were stratified by BW and allocated randomly in each of two periods to one of six treatments arranged in a two × three factorial arrangement. Treatments consisted of recommended (RM; 46.8%) or low (LM; 39.7%) moisture at baling after either no slurry application (NS), slurry application to stubble immediately after removal of the previous cutting (S0), or slurry application 14 d after removal of the previous cutting (S14). Silages were chopped through a commercial straw chopper, packed into plastic trash cans, and then offered to ewes within 4 d of chopping. Period 1 of the intake and digestion study consisted of a 14-d adaptation followed by a 7-d fecal collection period. Period 2 followed period 1 after a 4-d rest and consisted of an 11-d adaptation followed by 7 d of fecal collection. Ewes were housed individually in 1.4 × 4.3-m pens equipped with rubber mat flooring. Feces were swept from the floor twice daily, weighed, and dried at 50 °C. Ewes had ad libitum access to water and were offered chopped silage for a minimum of 10% refusal (DM). Blood samples were collected immediately prior to feeding, and 4 and 8 h after feeding on the day prior to the end of each period. Organic matter intake (g/kg BW) and OM digestibility tended (P < 0.10) to be, and digestible OM intake (g/kg BW) was reduced by slurry application. Lymphocytes (% of total white blood cells) were greater (P < 0.05) from LM vs. RM and from NS vs. S0 and S14. Red blood cell concentrations were greater (P < 0.05) from S14 vs. S0 and from S0 and S14 vs. NS. Serum urea N concentrations did not differ (P > 0.17) across treatments. Therefore, moisture concentration of alfalfa silage within the range used in this study may not affect voluntary intake or digestibility, but slurry application may have an effect on digestible OM intake. Also, moisture concentration of alfalfa silage and time of dairy slurry application may affect specific blood hemograms.

Effects of dairy slurry on silage fermentation characteristics and nutritive value of alfalfa.

Dairy producers frequently ask questions about the risks associated with applying dairy slurry to growing alfalfa (Medicago sativa L.). Our objectives were to determine the effects of applying dairy slurry on the subsequent nutritive value and fermentation characteristics of alfalfa balage. Dairy slurry was applied to 0.17-ha plots of alfalfa; applications were made to the second (HARV1) and third (HARV2) cuttings during June and July of 2012, respectively, at mean rates of 42,400 ± 5271 and 41,700 ± 2397 L/ha, respectively. Application strategies included (1) no slurry, (2) slurry applied directly to stubble immediately after the preceding harvest, (3) slurry applied after 1 wk of post-ensiled regrowth, or (4) slurry applied after 2 wk of regrowth. All harvested forage was packaged in large, rectangular bales that were ensiled as wrapped balage. Yields of DM harvested from HARV1 (2,477 kg/ha) and HARV2 (781 kg/ha) were not affected by slurry application treatment. By May 2013, all silages appeared to be well preserved, with no indication of undesirable odors characteristic of clostridial fermentations. Clostridium tyrobutyricum, which is known to negatively affect cheese production, was not detected in any forage on either a pre- or post-ensiled basis. On a pre-ensiled basis, counts for Clostridium cluster 1 were greater for slurry-applied plots than for those receiving no slurry, and this response was consistent for HARV1 (4.44 vs. 3.29 log10 genomic copies/g) and HARV2 (4.99 vs. 3.88 log10 genomic copies/g). Similar responses were observed on a post-ensiled basis; however, post-ensiled counts also were greater for HARV1 (5.51 vs. 5.17 log10 genomic copies/g) and HARV2 (5.84 vs. 5.28 log10 genomic copies/g) when slurry was applied to regrowth compared with stubble. For HARV2, counts also were greater following a 2-wk application delay compared with a 1-wk delay (6.23 vs. 5.45 log10 genomic copies/g). These results suggest that the risk of clostridial fermentations in alfalfa silages is greater following applications of slurry. Based on pre- and post-ensiled clostridial counts, applications of dairy slurry on stubble are preferred (and less risky) compared with delayed applications on growing alfalfa.

Fall-grown oat to extend the fall grazing season for replacement dairy heifers.

Our objective was to assess the pasture productivity and forage characteristics of 2 fall-grown oat (Avena sativa L.) cultivars, specifically for extending the grazing season and reducing reliance on harvested forages by replacement dairy heifers. A total of 160 gravid Holstein heifers (80 heifers/yr) were stratified by weight, and assigned to 1 of 10 identical research pens (8 heifers/pen). Initial body weights were 480 ± 43.5 kg in 2011 and 509 ± 39.4 kg in 2012. During both years of the trial, four 1.0-ha pasture replicates were seeded in August with Ogle oat (Schumitsch Seed Inc., Antigo, WI), and 4 separate, but similarly configured, pasture replicates were seeded with Forage Plus oat (Kratz Farms, Slinger, WI). Heifer groups were maintained as units, assigned to specific pastures, and then allowed to graze fall-oat pastures for 6h daily before returning to the barn, where they were offered a forage-based basal total mixed ration. Two heifer groups were retained in confinement (without grazing) as controls and offered the identical total mixed ration as pasture groups. During 2011, available forage mass increased with strong linear and quadratic effects for both cultivars, peaking at almost 9 Mg/ha on October 31. In contrast, forage mass was not affected by evaluation date in 2012, remaining ≤ 2,639 kg/ha across all dates because of droughty climatic conditions. During 2012, Ogle exhibited greater forage mass than Forage Plus across all sampling dates (2,678 vs. 1,856 kg/ha), largely because of its more rapid maturation rate and greater canopy height. Estimates of energy density for oat forage ranged from 59.6 to 69.1% during 2011, and ranged narrowly from 68.4 to 70.4% during 2012. For 2011, responses for both cultivars had strong quadratic character, in which the most energy-dense forages occurred in mid November, largely due to accumulation of water-soluble carbohydrates that reached maximum concentrations of 18.2 and 15.1% for Forage Plus and Ogle, respectively. Across the 2-yr trial, average daily gain for grazing heifer groups tended to be greater than heifers remaining in confinement (0.85 vs. 0.74 kg/d), but both management strategies produced weight gains within reasonable proximity to normal targets for heifers in this weight range. Fall-grown oat should be managed as stockpiled forage for deferred grazing, and good utilization of fall-oat forage can be accomplished by a one-time removal of standing forage, facilitated by a single lead wire advanced daily to prevent waste.

Unique interrelationships between fiber composition, water-soluble carbohydrates, and in vitro gas production for fall-grown oat forages.

Sixty samples of 'ForagePlus' oat were selected from a previous plot study for analysis of in vitro gas production (IVGP) on the basis of 2 factors: (1) high (n=29) or low (n=31) neutral detergent fiber (NDF; 62.7±2.61 and 45.1±3.91%, respectively); and (2) the range of water-soluble carbohydrates (WSC) within the high- and low-NDF groups. For the WSC selection factor, concentrations ranged from 4.7 to 13.4% (mean=7.9±2.06%) and from 3.5 to 19.4% (mean=9.7±4.57%) within high- and low-NDF forages, respectively. Our objectives were to assess the relationships between IVGP and various agronomic or nutritional characteristics for high- and low-NDF fall-oat forages. Cumulative IVGP was fitted to a single-pool nonlinear regression model: Y=MAX × (1 - e ([-)(K)(× (t - lag)])), where Y=cumulative gas produced (mL), MAX=maximum cumulative gas produced with infinite incubation time (mL), K=rate constant, t=incubation time (h), and lag=discrete lag time (h). Generally, cumulative IVGP after 12, 24, 36, or 48h within high-NDF fall-oat forages was negatively correlated with NDF, hemicellulose, lignin, and ash, but positively correlated with WSC, nonfiber carbohydrate (NFC), and total digestible nutrients (TDN). For low-NDF fall-grown oat forages, IVGP was positively correlated with growth stage, canopy height, WSC, NFC, and TDN; negative correlations were observed with ash and crude protein (CP) but not generally with fiber components. These responses were also reflected in multiple regression analysis for high- and low-NDF forages. After 12, 24, or 36h of incubation, cumulative IVGP within high-NDF fall-oat forages was explained by complex regression equations utilizing (lignin:NDF)(2), lignin:NDF, hemicellulose, lignin, and TDN(2) as independent variables (R(2)≥0.43). Within low-NDF fall-grown oat forages, cumulative IVGP at these incubation intervals was explained by positive linear relationships with NFC that also exhibited high coefficients of determination (R(2)≥0.75). Gas production was accelerated at early incubation times within low-NDF forages, specifically in response to large pools of WSC that were most likely to be present as forages approached boot stage by late-fall.

Technical note: whole-pen assessments of nutrient excretion and digestibility from dairy replacement heifers housed in sand-bedded freestalls.

Our objectives were to describe and test refined procedures for quantifying excreta produced from whole pens of dairy heifers. Previous research efforts attempting to make whole-pen measurements of excreta output have been complicated by the use of organic bedding, which requires cumbersome analytical techniques to quantify excreta apart from the bedding. Research pens equipped with sand-bedded freestalls offer a unique opportunity for refinement of whole-pen fecal collection methods, primarily because sand-bedded freestall systems contain no organic bedding; therefore, concentrations of ash within the manure, sand, and feces can be used to correct for contamination of manure by sand bedding. This study was conducted on a subset of heifers from a larger production-scale feeding trial evaluating ensiled eastern gamagrass [Tripsacum dactyloides (L.) L.] haylage (EGG) that was incorporated into a corn silage/alfalfa haylage-based blended diet at rates of 0, 9.1, 18.3, or 27.4% of total DM. The diet without EGG also was offered on a limit-fed basis. Eighty Holstein dairy heifers were blocked (heavy weight, 424 ± 15.9 kg; light weight, 324 ± 22.4 kg) and then assigned to 10 individual pens containing 8 heifers/pen. One pen per block was assigned to each of the 5 research diets, and whole-pen fecal collections were conducted twice for each pen. Grab fecal samples also were gathered from individual heifers within each pen, and subsequent analysis of these whole-pen composites allowed reasonable estimates of OM and NDF excreta output. Under the conditions of our experimental design, pooled SEM for the excreta DM, OM, NDF, and NDF (ash corrected) output were 0.113, 0.085, 0.093, and 0.075 kg·heifer(-1)·d(-1), respectively. For DM excretion, this represented about one-third of the SEM reported for previous whole-pen collections from bedded-pack housing systems. Subsequent calculations of apparent DM and OM digestibilities indicated that the technique was sensitive, and linear trends (P ≤ 0.027) associated with the inclusion rates of EGG within the diet were detected. This technique allows estimation of apparent diet digestibilities on multiple animals simultaneously, thereby mitigating the need for isolating individual animals to obtain digestibility coefficients. The approach appears viable but requires hand labor for collections of multiple pens and thorough mixing of large volumes of manure as well as analytical corrections for sand ingested by lounging heifers.

Storage characteristics, nutritive value, energy content, and in vivo digestibility of moist, large rectangular bales of alfalfa-orchardgrass hay treated with a propionic acid-based preservative.

Unstable weather, poor drying conditions, and unpredictable rainfall events often place valuable hay crops at risk. Recent research with large round bales composed of alfalfa (Medicago sativa L.) and orchardgrass (Dactylis glomerata L.) has shown that these large-bale packages are particularly sensitive to spontaneous heating and dry matter (DM) losses, as well as other undesirable changes with respect to forage fiber, protein, and energy density. Various formulations of organic acids have been marketed as preservatives, normally for use on hays that are not desiccated adequately in the field to facilitate safe bale storage. Our objectives for this study were to (1) evaluate the efficacy of applying a commercial (buffered) propionic acid-based preservative at 3 rates (0, 0.6, and 1.0% of wet-bale weight) to hays baled at 3 moisture concentrations (19.6, 23.8, and 27.4%) on the subsequent storage characteristics and poststorage nutritive value of alfalfa-orchardgrass forages packaged in large rectangular (285-kg) bales, and then (2) evaluate the in vivo digestibility of these hays in growing lambs. Over a 73-d storage period, the preservative was effective at limiting spontaneous heating in these hays, and a clear effect of application rate was observed for the wettest (27.4%) bales. For drier hays, both acid-application rates (1.0 and 0.6%) yielded comparable reductions in heating degree days >30°C relative to untreated controls. Reductions in spontaneous heating could not be associated with improved recovery of forage DM after storage. In this study, most changes in nutritive value during storage were related to measures of spontaneous heating in simple linear regression relationships; this suggests that the modest advantages in nutritive value resulting from acid treatment were largely associated with perturbations of normal heating patterns during bale storage. Although somewhat erratic, apparent digestibilities of both DM (Y=-0.0080x + 55.6; R(2)=0.45) and organic matter (Y=-0.0085x + 55.5; R(2)=0.53) evaluated in growing lambs were also directly related to heating degree days in simple linear relationships. Based on these data, applying propionic acid-based preservatives to large rectangular bales is likely to provide good insurance against spontaneous heating during storage, as well as modest benefits with respect to nutritive value and digestibility.

Using eastern gamagrass to construct diets that limit intake and caloric density for dairy replacement heifers.

Previous research has shown that eastern gamagrass (EGG; Tripsacum dactyloides L.) will survive winter climatic conditions common throughout central Wisconsin, and will produce yields of dry matter (DM) ranging approximately from 7,000 to 10,000 kg/ha annually when managed with a 1-cut harvest system. The objective of this research was to determine whether the fibrous nature of this perennial warm-season grass could be effective in reducing the caloric density and DMI of corn silage/alfalfa haylage diets for replacement dairy heifers. A total of 120 Holstein dairy heifers were blocked by body weight (heavy, 424 ± 15.9 kg; medium, 369 ± 11.8 kg; light, 324 ± 22.4 kg), and then assigned to 15 individual pens containing 8heifers each. Eastern gamagrass forage was harvested, ensiled, and subsequently incorporated into blended corn silage/alfalfa haylage diets at rates of 0, 9.1, 18.3, or 27.4% of the total dietary DM (EGG0, EGG9, EGG18, and EGG27, respectively). These diets were offered during a 105-d evaluation period for ad libitum intake; however, the EGG0 diet also was offered on a limit-fed basis (LF), which was set at 85% of the voluntary intake of EGG0. Serial additions of EGG increased concentrations of neutral detergent fiber in blended diets from 39.6 (EGG0) to 48.7% (EGG27), and simultaneously reduced corresponding estimates of total digestible nutrients (TDN) from 68.2 to 61.3%, and net energy for gain from 1.07 to 0.83 Mcal/kg. Dry matter intakes for all diets offered ad libitum were greater than observed for LF (9.06 vs. 8.07 kg/d); however, DM intakes for diets containing EGG were reduced relative to EGG0 (9.40 vs. 8.94 kg/d). Similarly, intakes of TDN were greater for diets offered for ad libitum intake than for LF (5.84 vs. 5.50 kg/d); however, inclusion of EGG reduced TDN intakes relative to EGG0 (6.41 vs. 5.65 kg/d). This reduction was explained by both linear and quadratic effects of the inclusion rate of EGG in the diet. Over the 105-d trial, total weight gains ranged from 89 kg (0.85 kg/d) for heifers offered EGG27 up to 114 kg (1.09 kg/d) for those offered EGG0. Performance was similar between heifers offered EGG27 and LF diets (0.85 vs. 0.88 kg/d). Eastern gamagrass haylage proved to be a completely nonsortable additive within corn silage/alfalfa haylage diets. It also was effective in limiting the caloric density and DM intake of these diets, as well as undesirable weight gains by dairy heifers.

Effects of a propionic acid-based preservative on storage characteristics, nutritive value, and energy content for alfalfa hays packaged in large round bales.

During 2009 and 2010, alfalfa (Medicago sativa L.) hays from 2 cuttings harvested from the same field site were used to evaluate the effects of a propionic acid-based preservative on the storage characteristics and nutritive value of hays stored as large round bales. A total of 87 large round bales (diameter = 1.5m) were included in the study; of these, 45 bales served as controls, whereas 42 were treated with a commercial propionic acid-based preservative at mean application rates of 0.5±0.14 and 0.7±0.19% of bale weight, expressed on a wet (as is) or dry matter basis, respectively. Initial bale moisture concentrations ranged from 10.2 to 40.4%. Internal bale temperatures were monitored daily during an outdoor storage period, and heating characteristics were summarized for each bale as heating degree days (HDD) >30°C. For acid-treated bales, the regression relationship between HDD and initial bale moisture was best fitted to a quadratic model in which the linear term was dropped to improve fit (Y=2.02x(2) - 401; R(2)=0.77); control hays were best fitted to a nonlinear model in which the independent variable was squared [Y=4,112 - (4,549×e(-0.000559x*x)); R(2)=0.77]. Based on these regressions, acid-treated bales accumulated more HDD than control hays when the initial bale moisture was >27.7%; this occurred largely because acid treatment tended to prolong active heating relative to control hays. Linear regressions of recoveries of dry matter on HDD did not differ on the basis of treatment, yielding a common linear relationship of Y=-0.0066x+96.3 (R(2)=0.75). Regressions relating changes (post-storage - pre-storage) in concentrations of several nutritional components (neutral detergent fiber, lignin, ash, crude protein, and total digestible nutrients) with HDD for acid-treated hays typically exhibited more inflection points or were higher-ordered polynomial regressions than those of control hays. These more complex responses probably reflected the perturbation of normal heating patterns following acid treatment; however, overall effects on post-storage nutritive value were relatively limited in scope. The potential to improve nutritive value relative to cost for these large round bales was not especially favorable, and hay producers may find that diligence to achieve adequate field desiccation before baling, or use of oxygen-exclusion methods, such as wrapping in plastic, may be better alternatives for preserving moist hays.

Technical note: a near-infrared reflectance spectroscopy technique to predict particle size of starch within corn silage.

Starch particle size characteristics of 81 diverse corn silage samples, which included 27 combinations of hybrid, planting dates, and harvest dates subjected to 3 different degrees of kernel processing, were determined via vertical shaking through 9 screens with nominal square apertures of 19.0, 13.2, 9.5, 6.7, 4.75, 3.35, 2.36, 1.18, and 0.6 mm and a pan. Starch content of dry matter remaining on each screen and on the pan for each corn silage was determined, and geometric mean particle size (GMPS, mum), starch particles(SP)/g, starch surface area (SSA, cm(2)/g), kernel processing score (KPS), % starch < 4.75 mm, and the percentage of total starch remaining on each screen of the vertical shakers were calculated. Near-infrared reflectance spectra were obtained by scanning 3 types of samples: 1 mm of dried ground corn silage; whole undried, unground corn silage; and undried, unground corn silage that passed through a 19-mm screen. Calibrations to predict GMPS, SP, SSA, and KPS characteristics of corn silage starch were attempted from each spectral origin. Calibrations to predict GMPS, SP, SSA, KPS, and the percentage of total starch retained on screens of the vertical shaker was unattainable (R(2) < 0.45) using spectra obtained from 1 mm of dried ground corn silage or whole undried, unground corn silage. However, reasonable near-infrared reflectance spectra equations (R(2) > 0.81) for GMPS, SSA, and KPS were attained using spectra from undried, unground corn silage that passed through a 19-mm screen. This technique holds promise as a rapid and efficient method to determine particle size characteristics of starch within corn silage.