Evidence Of Carry-over Effects Research Articles

The Effects of Mild Disturbances on Sleep Behaviour in Laying Hens

The positive welfare of commercial animals presents many benefits, making the accurate assessment of welfare important. Assessments frequently use behaviour to determine welfare state; however, nighttime behaviours are often ignored. Sleep behaviour may offer new insights into welfare assessments. This study aimed to establish a baseline for sleep behaviour in laying hens and to then apply mild short-term disturbances and observe the subsequent effects. Twelve laying hens were divided into four batches and were surgically implanted with electroencephalogram (EEG) devices to record their brain activity. The batches were subjected to undisturbed, disturbed and recovery types of nights. Disturbed nights consisted of systematic sequences of disturbance application (wind, 90 dB noise or 20 lux light) applied one at a time for 5 min every 30 min from 21:00 to 03:00 (lights off period: 19:00-05:00). Sleep state was scored using EEG data and behaviour data from infrared cameras. Over all the types of night hens engaged in both SWS (58%) and REM sleep (18%) during lights off. When applied, the disturbances were effective at altering the amounts of wakefulness and SWS (Time × Type of Night, p < 0.001, p = 0.017, respectively), whereas REM sleep was unaltered (p = 0.540). There was no evidence of carry-over effects over the following day or night. Laying hens may be resilient to short-term sleep disruption by compensating for this in the same night, suggesting that these disturbances do not impact their long-term welfare (i.e., over days). Sleep behaviour potentially offers a unique means of assessing an aspect of animal welfare that, to date, has been poorly studied.

A sensorimotor approach to disfluency adaptation in typically fluent adults

Delaying auditory feedback (DAF) during speech is a potent auditory perturbation that can interrupt and prolong syllables and words, even though these disfluencies rarely affect typical speakers under non-altered feedback (NAF). Adaptation or compensation to auditory perturbations has been shown in typical speakers through a gradual reduction in the amplitude of formant shift responses. However, despite considerable research on DAF, it is still not known whether typical speakers adapt to DAF. In this study, we tested whether a comparable form of adaptation occurs with DAF in typical speakers as shown by a reduction in altered feedback disfluencies (AFD) during repeated consecutive readings, after a pause between readings and to a novel reading. We then tested for carryover effects after a single DAF exposure. A significant decrease in AFD rate was observed in 38 speakers that was sustained after a pause and for a novel reading. The adaptation effect extended to articulation rate (syllables/sec) in that rate increased for all speakers across readings. Evidence for carryover effects was inconclusive. By showing that typical speakers can adapt to DAF, the findings support a sensorimotor approach to auditory perturbation adaptation achieved through motor practice.

The Effects of Aquatic Watsu Therapy on Gross Motor Performance and Quality of Life for Children with Cerebral Palsy

The purpose of this study was to compare the effects of Immersion and Watsu® therapy protocols on children with cerebral palsy. Gross motor function (GMF), spasticity (MAS), and Quality of Life (QoL) parameters of twenty-three children (age: 7.5±2.8, BMI: 17±3.7) were measured. Subjects received Watsu® therapy and Immersion protocols in Watsu-Immersion (W-I, n=12) and Immersion-Watsu (I-W n=11) groups in different periods based on a crossover design. The subjects received sessions for 30 minutes twice a week during 10-week experimental periods. The results were investigated with independent sample t-test and Wilcoxon rank-sum tests, which showed that there was no evidence of carryover effects in GMFM 88 and QoL. In contrast, Watsu® improved GMFM 88 (p&lt;0.05), Quality of Life (p&lt;0.05), and MAS-Upper spasticity (p&lt;0.05), scores significantly compared to immersion. The current results demonstrated the specific benefits of Watsu® therapy on children with CP, confirming the previous anecdotal reports. We recommend Watsu® as a safe and well-accepted complementary intervention for the management of CP.

Selective Inhibition of PDE4B Reduces Binge Drinking in Two C57BL/6 Substrains.

Cyclic AMP (cAMP)-dependent signaling is highly implicated in the pathophysiology of alcohol use disorder (AUD), with evidence supporting the efficacy of inhibiting the cAMP hydrolyzing enzyme phosphodiesterase 4 (PDE4) as a therapeutic strategy for drinking reduction. Off-target emetic effects associated with non-selective PDE4 inhibitors has prompted the development of selective PDE4 isozyme inhibitors for treating neuropsychiatric conditions. Herein, we examined the effect of a selective PDE4B inhibitor A33 (0–1.0 mg/kg) on alcohol drinking in both female and male mice from two genetically distinct C57BL/6 substrains. Under two different binge-drinking procedures, A33 pretreatment reduced alcohol intake in male and female mice of both substrains. In both drinking studies, there was no evidence for carry-over effects the next day; however, we did observe some sign of tolerance to A33’s effect on alcohol intake upon repeated, intermittent, treatment (5 injections of 1.0 mg/kg, every other day). Pretreatment with 1.0 mg/kg of A33 augmented sucrose intake by C57BL/6NJ, but not C57BL/6J, mice. In mice with a prior history of A33 pretreatment during alcohol-drinking, A33 (1.0 mg/kg) did not alter spontaneous locomotor activity or basal motor coordination, nor did it alter alcohol’s effects on motor activity, coordination or sedation. In a distinct cohort of alcohol-naïve mice, acute pretreatment with 1.0 mg/kg of A33 did not alter motor performance on a rotarod and reduced sensitivity to the acute intoxicating effects of alcohol. These data provide the first evidence that selective PDE4B inhibition is an effective strategy for reducing excessive alcohol intake in murine models of binge drinking, with minimal off-target effects. Despite reducing sensitivity to acute alcohol intoxication, PDE4B inhibition reduces binge alcohol drinking, without influencing behavioral sensitivity to alcohol in alcohol-experienced mice. Furthermore, A33 is equally effective in males and females and exerts a quantitatively similar reduction in alcohol intake in mice with a genetic predisposition for high versus moderate alcohol preference. Such findings further support the safety and potential clinical utility of targeting PDE4 for treating AUD.

No evidence for carryover effect in tree rings based on a pulse-labelling experiment on Juniperus communis in South Germany

A clear carryover effect of tree-ring formation was not detected based on a pulse-labelling experiment conducted on Juniperus communis in South Germany. The inherent linkage between photosynthesis and the formation of wood is important for the understanding of relationships between tree-ring series and climate/environmental data in dendroclimatology studies. However, it is impossible to reach a mechanistic procedure of tree stem radial growth depending on its carbon balance from a traditional statistical point of view alone. Pulse labelling experiment with stable carbon isotope (13CO2) has provided innovative insights into the fate of recently assimilated carbon in organs and carbon-containing compounds. In this study, we conducted an in situ pulse labelling experiment on 27 July 2016 to examine the response of tree ring and different-aged needles to short-term elevated 13CO2 of a juniper shrub growing on a heathland in South Germany. New and old needles from four expositions were sampled before and after the experiment. A wood segment was taken from the main branch and stable carbon isotope composition (δ13C) was analysed at an intra-annual time scale. Before the experiment, the mean δ13C was − 26.8 ± 0.4‰ (mean ± standard deviation) for both needle ages, while woody tissue showed about 3‰ higher δ13C compared to needles. Substantial enriched 13C was detected in the needles after the experiment. New needles showed significant higher δ13C than the old ones 1–7 days after the experiment. Significant enriched δ13C was detected in the wood from 35 to 61% of the annual tree ring in 2016, indicating that the short-term enrichment of 13C can affect wood formation for a large section. No enhancement in 13C signal appeared in the tree ring of the subsequent year 2017, suggesting the absence of a carryover effect. Wood formation did not reply on the carbohydrates stored even 1 year before and thus tree-ring ecophysiological modelling as well as dendrochronological studies should therefore benefit from such result.

Sub-Perception and Supra-Perception Spinal Cord Stimulation in Chronic Pain Syndrome: A Randomized, Semi-Double-Blind, Crossover, Placebo-Controlled Trial.

Background: The introduction of modern sub-perception modalities has improved the efficacy of spinal cord stimulation (SCS) in refractory pain syndromes of the trunk and lower limbs. The objective of this study was to evaluate the effectiveness of low and high frequency SCS among patients with chronic pain. Material and methods: A randomised, semi-double-blind, placebo controlled, four period (4 × 2 weeks) crossover trial was conducted from August 2018 to January 2020. Eighteen patients with SCS due to failed back surgery syndrome and/or complex regional pain syndrome were randomised to four treatment arms without washout periods: (1) low frequency (40-60 Hz), (2) 1 kHz, (3) clustered tonic, and (4) sham SCS (i.e., placebo). The primary outcome was pain scores measured by visual analogue scale (VAS) preoperatively and during subsequent treatment arms. Results: Pain scores (VAS) reported during the preoperative period was M (SD) = 8.13 (0.99). There was a 50% reduction in pain reported in the low frequency tonic treatment group (M (SD) = 4.18 (1.76)), a 37% reduction in the 1 kHz treatment group (M (SD) = 5.17 (1.4)), a 34% reduction in the clustered tonic settings group (M (SD) = 5.27 (1.33)), and a 34% reduction in the sham stimulation group (M (SD) = 5.42 (1.22)). The reduction in pain from the preoperative period to the treatment period was significant in each treatment group (p < 0.001). Overall, these reductions were of comparable magnitude between treatments. However, the modality most preferred by patients was low frequency (55% or 10 patients). Conclusions: The pain-relieving effects of SCS reached significance and were comparable across all modes of stimulation including sham. Sub-perception stimulation was not superior to supra-perception. SCS was characterised by a high degree of placebo effect. No evidence of carryover effect was observed between subsequent treatments. Contemporary neuromodulation procedures should be tailored to the individual preferences of patients.

The Role of Protein Quality and Quantity in Promoting Fullness and Reducing Energy Intake (P21-056-19)

Abstract Objectives A breakfast containing eggs (EB) can potentially provide a greater quantity of high quality protein. We previously reported (Study 1) that EB induces greater satiety and reduces lunch time energy intake (LTEI) compared to a breakfast without eggs (CB), but of equal energy density. However, this EB had a greater amount of protein. Hence, in Study 2 we compared EB and CB with equal energy density and protein quantity, but of differing protein quality. EB (higher protein quality) increased fullness, but did not reduce LTEI. For this study (Study 3), we used the breakfasts from Study 2, to compare the effect on fullness and LTEI, in subjects in negative energy balance, due to a weight loss diet. Methods In a crossover study, 30 sedentary women (18–44 y old) with overweight or obesity (BMI 25–39.9 kg/m2) but otherwise healthy, received baseline measurements and weight loss counseling. They visited the clinic daily to receive in random order either EB or CB each for one week. The two breakfasts were similar in energy density and macronutrients but differed in protein quality (mainly egg protein vs wheat protein). On days 7 and 14, satiety questionnaires using visual analog scales (VAS) were offered at 7 time points (−30, 30, 60, 90, 120, 180, and 240 min), before and after the breakfasts. An ad libitum lunch was offered on days 7 and 14 and the energy intake was covertly recorded. Results Area under the curve (AUC) of each factor were compared between 2 breakfasts using a linear mixed effects model, adjusting for subject and visit. EB showed significantly more fullness (P = 0.038). The differences between the AUCs between two breakfasts for hunger, thirst, nausea, and the amount that could be further eaten were not significantly different. There was no significant difference in LTEI between EB and CB. There was no evidence of carry-over effects. Conclusions A synthesis of the 3 studies indicates that when an EB is superior in protein quality and quantity, it can increase fullness and reduce LTEI. When the EB is higher in protein quality, but matched for protein quantity, it still increases fullness, but does not reduce LTEI. This insight could help in developing a weight loss diet plan, when increasing fullness and reducing energy intake are important considerations. Funding Sources American Egg Board/Egg Nutrition Center.

Overwintering habitat links to summer reproductive success: intercontinental carry-over effects in a declining migratory bird revealed using stable isotope analysis

ABSTRACTCapsule: Breeding success in female Pied Flycatchers Ficedula hypoleuca is related to isotopic signature of feathers grown in Africa, suggesting wintering habitat links to breeding performance 5000 km away.Aims: Better understanding of inter-seasonal carry-over effects is a research priority, especially for declining migrants. We use stable isotope analysis to relate Pied Flycatcher winter habitat to summer reproductive success.Methods: Flycatchers were captured in three UK woodlands in 2013–15. An Africa-grown tertial was trimmed and analysed using Isotope Ratio Mass Spectrometry to quantify Nitrogen-15 (δ15N) and Carbon-13 (δ13C). In total, 135 samples were taken from 80 individuals.Results: Wintering δ15N and δ13C differed significantly between years. δ13C correlated with lay date, such that birds with lower carbon levels (indicative of more mesic habitat) bred earlier. There was a significant correlation between wintering δ13C and productivity after allowing for year, site and lay date; birds with low δ13C were more successful. This suggests δ13C links productivity directly as well as indirectly through phenological effects. δ15N did not relate to phenology or productivity.Conclusion: This is the first evidence of carry-over effects between geographical regions for a European passerine. Conservation measures should focus on all aspects of seasonal cycles, not just breeding grounds.

Executive Attention as a Predictor of Distracted Driving Performance

The present study empirically examines the role of executive attention as a predictor of distracted driving performance. As ADHD is an attention-related disorder, we hypothesized that ADHD individuals would show poorer executive attention compared to individuals from a community sample. Second, we predicted that individuals with poorer executive attention would show riskier driving as measured by a self-report questionnaire and simulated driving trials. One-hundred thirteen individuals with and without ADHD completed an executive attention task, a driving behavior questionnaire, and three (pre-distracted, distracted, and post-distracted) simulated driving routes. Results showed that executive attention is associated with braking response times during distracted driving trials. There was also evidence for carryover effects of distraction, with participants showing slower braking response times even after the distracting task was discontinued. Further research is needed to clarify the relationship between ADHD and executive attention.

When Siberia came to the Netherlands: the response of continental black-tailed godwits to a rare spring weather event.

1. Extreme weather events have the potential to alter both short- and long-term population dynamics as well as community- and ecosystem-level function. Such events are rare and stochastic, making it difficult to fully document how organisms respond to them and predict the repercussions of similar events in the future. 2. To improve our understanding of the mechanisms by which short-term events can incur long-term consequences, we documented the behavioural responses and fitness consequences for a long-distance migratory bird, the continental black-tailed godwit Limosa limosa limosa, resulting from a spring snowstorm and three-week period of record low temperatures. 3. The event caused measurable responses at three spatial scales - continental, regional and local - including migratory delays (+19 days), reverse migrations (>90 km), elevated metabolic costs (+8·8% maintenance metabolic rate) and increased foraging rates (+37%). 4. There were few long-term fitness consequences, however, and subsequent breeding seasons instead witnessed high levels of reproductive success and little evidence of carry-over effects. 5. This suggests that populations with continued access to food, behavioural flexibility and time to dissipate the costs of the event can likely withstand the consequences of an extreme weather event. For populations constrained in one of these respects, though, extreme events may entail extreme ecological consequences.

Foraging capability of extraradical mycelium of arbuscular mycorrhizal fungi to soil phosphorus patches and evidence of carry-over effect on new host plant

The foraging capability of extraradical mycelium (ERM) of arbuscular mycorrhizal (AM) fungi to high soil phosphorus (P) supply and carry-over effect of ERM on new host plants remain to be explored. This study investigated whether the inhibition of ERM at high P supply varies between fungal species and whether ERM have carry-over effects when they encounter new hosts. A two-compartment device was designed to spatially separate hyphae from host roots and the carry-over effect was investigated using two-stage growth experiments. P fertilizers were applied only to hyphal compartments at a gradient of 0, 35, 150, 300, or 1,000 mg P∙kg−1. Maize plants were inoculated with Glomus mosseae, Rhizophagus irregularis or remained uninoculated. The biomass of G. mosseae in the hyphal compartment was on average higher than that of R. irregularis and the difference was greater at the second harvest. The biomass of G. mosseae increased slightly at P35 and thereafter decreased significantly with increasing P level. The biomass of R. irregularis increased slightly at P35 and did not differ significantly at higher P levels from that at P0. In general, inoculation significantly increased the biomass and P uptake of donor and receiver plants. The mycorrhizal growth responsiveness (MGR) and mycorrhizal phosphorus responsiveness (MPR) of both donor and receiver plants decreased at higher P supply level. The ratios of MGR and MPR between donor and receiver plants were close to 1 for both fungal species and were independent of P level. Inoculation with R. irregularis significantly enhanced root P uptake efficiency and alkaline phosphatase activity (ALP) in both donor and receiver plants across all P levels. Our results demonstrate that fungal species differ in their responses to soil P availability. R. irregularis was less sensitive to soil P gradients, and compensated for growth promotion and P uptake of the host by increasing ALP and root P uptake efficiency. The carry-over effects of ERM imply that the maintenance of mycorrhizal networks is of particular importance in intensive agricultural ecosystems.

Dynamics of ungulates in relation to climatic and land use changes in an insularized African savanna ecosystem

Land use change and human population growth are accelerating the fragmentation and insularization of wildlife habitats worldwide. The conservation and management of wildlife in the resultant ‘island’ ecosystems in the context of global warming is challenging due to the isolation and reduced size of the ecosystems and hence the scale over which ecosystem processes can operate. We analyzed trends in numbers of nine large herbivores in Kenya’s Lake Nakuru National Park to understand how rainfall and temperature variability, surrounding land use changes, and boundary fencing affected wildlife population dynamics inside the park during 1970–2011. Buffalo, zebra and Thomson’s gazelle numbers increased persistently. Grant’s gazelle and impala increased initially then gradually declined. Waterbuck and warthog numbers progressively declined to levels that potentially threatened their local population persistence. The total biomass of ungulates tripled from 1970 to 2011, with buffalo replacing waterbuck as the predominant species in biomass. Increased competition from buffalo and zebra, heightened predation and illicit human harvests probably all contributed to the declines by waterbuck and warthog. Density-dependent limitation of population growth within the park confines was evident for buffalo, impala, eland, giraffe, Grant’s and Thomson’s gazelles. Fluctuations in the lake level related to varying rainfall affected changes in animal abundance through expansion of the lake area and flooding of grasslands bordering the lake. Unusually, the most stressful conditions were associated with high water levels following high rainfall. There was also evidence of carry-over effects from prior habitat conditions affecting all species. The relatively stable populations of all species except warthog and waterbuck demonstrate the remarkable capacity of this small, insularized park to retain viable populations of most of the large herbivores, without much management intervention.

Electroanalytical determination of carbendazim by square wave adsorptive stripping voltammetry with a multiwalled carbon nanotubes modified electrode

A preconcentrating/voltammetric multiwalled carbon nanotube modified glassy carbon electrode (MWCNT–GCE) has been developed for stripping analysis of carbendazim (Methyl Benzimidazol-2-yl Carbamate—MBC), based on dispersing MWCNT in water. The effect of experimental variables, such as the dispersion and loading of MWCNT, was assessed. A quasi-reversible behavior for MBC in acetic acid/acetate buffer 0.1 mol L−1 (pH 4.7) was verified and its high effective pre-concentration was attributed to the high adsorption capability and enormous surface area of the MWCNT. No evidence of carry-over effect, combined with the easiness of electrode preparation, led to the development of a highly sensitive and reliable method with an experimental work range from 0.256 to 3.11 µmol L−1 with a detection limit of 10.5 ppb for a short (60 s) accumulation period. Measurement of MBC in a river water sample was demonstrated. The accuracy of the method for real sample analysis was assessed by estimating the apparent recovery (93 ± 2.9% and 86 ± 4.1% for 4.3 × 10−7 mol L−1) for a MBC spiked river water sample.

Speed/accuracy trade-offs in rapid simultaneous and sequential actions: Evidence for carryover effects

The idea of a neuromuscular synergy involving independent amplitude commands to the agonist and antagonist musculature is quite an appealing part of Plamondon's theory. One question that might be raised relates to the relative independence of the two “D” commands. Evidence is presented that suggests that the two commands might be related in sequential or simultaneous rapid aiming movements.

The Effects of Working in Pairs in Science Performance Assessments

Concerns about the effects of multiple-choice measures in traditional testing programs have led many educators and policymakers to suggest the use of alternative assessment methods. Some performance-based assessments require students to work in small collaborative groups as part of the test process. This study uses responses to hands-on science tasks at Grades 5 and 8 to examine whether the score a student earns while working with someone else is a truly independent assessment of that student's ability. We also explore whether working in pairs affects an individual's scores on subsequent tasks and whether these results are consistent across grade levels. Our analyses indicate that at Grades 5 and 8, work done with a partner should not be considered as an independent assessment of each student's ability. Some evidence of carry-over effects from working in pairs was found at each grade.

Children's sleep under the threat of attack by ballistic missiles.

The present paper reports on the influence of the Scud missile attacks during the Gulf War on the sleep of Israeli children. Two studies were performed. In the first, sleep habits and sleep disturbances of 61 (mean age 20 months) infants were assessed by questionnaires completed by their parents 5 months before the war and immediately after the end of the War. Comparison of pre- and post-war data revealed no major changes in sleep habits or in sleep quality. In the second study, sleep of 55 children was monitored at home by actigraphs during the last month of the War. All children were aroused during missile attacks, but returned to sleep immediately, with no evidence of carry-over effects once the 'all clear' sign was given. Comparison of sleep quality measures obtained during the War with those of age- and sex-matched children monitored a year before the war did not reveal any significant differences apart from the immediate response to the attack.

Isolation of protoplasts from loblolly pine needles and their flow-cytometric analysis for air pollution effects

Flow-cytometric analysis was used for the first time to determine the effects of air pollutants on plant biochemistry. The methods used were exploratory, but the results indicate that the technique is a valuable tool for researchers interested in the effects of air pollutants on vegetation. Evidence of carryover effects was seen in protoplasts isolated from new needles of 2-year-old loblolly pines ( Pinus taeda L.) which had been treated with ozone and simulated acidic rain the preceding year. Overall, staining of the protoplasts with constituent-specific fluorescent dyes showed decreased levels of fluorescence for esterase activity, protein, neutral lipids, and RNA in the ozone-treated samples compared with the charcoal-filtered or ambient samples. However, within pollutant treatments, low-level shifts in biochemistry were seen; for instance, simulated acidic rain at pH 5.0 ameliorated the effects of ozone on neutral lipids and RNA, and at pH 4.3 ameliorated the effects of ozone on protein. This research provides evidence at the biochemical level that pollutant treatments given in one year may be reflected in the succeeding year's growth. Image analysis of cross sections of fresh young pine needles supported the biochemical data.

Comprehensive evaluation of a model alcohol education curriculum.

A comprehensive evaluation was conducted of a representative alcohol education program designed to enhance knowledge and self-esteem, instill appropriate attitudes and teach the decision-making skills necessary for youth to make responsible decisions about the use of alcohol. The curriculum was implemented approximately as intended. The immediate, cumulative and longer-term effects of curriculum exposure on the variables thought to mediate alcohol use were modest or unsystematic. There was no consistent evidence of carryover effects from curriculum exposure on subsequent use of alcohol, cigarettes or other drugs. The curriculum was ineffective in attaining its goals.

High Levels of Grass Silage for Milk Production with No Grain, Medium, and High Grain Feeding. 1. Intake, Milk Production, and Body Weight Changes

Summary A three-lactation study with 18 Holstein and six Guernsey cows compared roughage intake, milk production, and weight changes on three levels of grain feeding (no grain, medium grain—1:6, and high grain—1:3) with maximum molasses-grass silage intake in addition to 5lb. good alfalfa hay. Pasture was supplied during the summer. For each pound of grain fed, roughage dry matter intake decreased 0.23lb. Average hay equivalent intake was 2.15lb. over all groups. Average 245-day 4% FCM production for 12 cows with an estimated potential of 8,830lb. was 6,490, 7,690, and 8,910lb. on the three rations; for 12 cows with an estimated potential of 10,930lb., production on the three rations was 8,080, 10,100, and 12,960lb., respectively. There was no evidence of carry-over effects in the reversal phase or cumulative effects in the continuous phase. There was a close relationship between potential producing ability of the cows and response, measured as the ratio of extra milk produced per pound of grain fed, comparing production on high grain with that on no grain (r = 0.88). Cows which had a 245-day potential of 8,900lb. FCM gave 0.58lb. milk per pound grain fed; whereas, those with a 245-day potential of 11,700lb. FCM gave 1.35lb. milk per pound grain fed. There was no evidence of diminishing returns in the high-potential cows, and relatively little for the lower-potential cows. Differences in rate of decline in production during the lactation were not apparent after the first 11 wk. of lactation, with little change after 6 wk. Though cows receiving no grain lost considerable weight from the first to the fourth month (−106lb.), compared with the high-grain cows (+16lb.), they gained more in the latter part of lactation (190 vs. 142lb.) ; however, they still did not regain their precalving weight in 50 wk. High-grain feeding may be profitable for cows with the potential for high production, but not for cows with lower potential, when fed large amounts of good-quality molasses-grass silage.