BIOMASS AND CHEMICAL RESPONSES OF Desmanthus spp. ACCESSIONS SUBMITTED TO WATER DEPRIVATION1

ABSTRACT Due to the predictions of climate change, there is a need to identify forage plants that can keep their productivity and nutritive value under hydric stress. The objective of this study was to evaluate the biomass and chemical responses of three Desmanthus spp. accessions under two water deprivation regimens (7 and 21-day). The experimental design used was randomized blocks in a 3 × 2 factorial arrangement (access and water deprivation) with four replications. There were reductions in the biomasses of leaf and stem fractions of 64% and 51%, respectively. The 43F accession showed greater leaf (1.86 g.plant-1) and stem (1.97 g.plant-1) biomasses under a total water restriction of 21 days, compared to the 89F and AusT accessions. In accessions 43F and 89F, there were carbohydrate accumulations in the leaves of 28% and 51%, respectively, under the 21-day water restriction, while AusT decreased 38% within a 7-day interval. Water deprivation affected the chemical characteristics of the accessions. Free proline was similar among accessions and accumulated 463% more in the leaves of plants submitted to 21-day water deprivation (90.22 mg.kg-1) compared to those subjected to water deprivation for 7 days (16.03 mg.kg-1). Proline and total soluble carbohydrate accumulation in 43F and 89F were insufficient to regulate crude protein, C content, and C:N ratios. These results demonstrate the variability in drought tolerance among accessions. Accessions 43F and 89F were more susceptible to 21-day water deprivation, while AusT showed greater drought tolerance.

Freely Moving Visual Discrimination Assay with Visiomode v2

The goal of this protocol is to assess visuomotor learning and motor flexibility in freely-moving mice, using the Visiomode touchscreen platform. Water-restricted mice first learn to associate touching a visual stimulus on the screen with a water reward. They then learn to discriminate between different visual stimuli on the touchscreen by nose-poking, before asked to switch their motor strategy to forelimb reaching. Version 1 of the protocol uses traditional water deprivation and water rewards in the task as a means of motivating mice to perform the task. Version 2 of the protocol uses Citric Acid for water restriction and sucrose as rewards in the task instead of the traditional water deprivation protocol.

Head-Restrained Lever Go/NoGo Visual Discrimination Task with Visiomode v2

This protocol is an adaptation of Michelle's lever Go/NoGo auditory discrimination task, which uses visual instead of auditory stimuli. Water-restricted, headplated mice learn to discriminate between a target and a distractor stimulus presented serially in pseudo-random order, pushing a lever to indicate when the target stimulus appears. The protocol is designed for across-learning recordings, and as such the inter-trial interval and stimulus interval remain constant throughout. Correction trials are also enabled throughout the duration of the protocol. The task in implemented in the Visiomode platform, using the lever apparatus for response input instead of the touchscreen. This protocol uses the Citric Acid Water Restriction protocol with sucrose rewards, instead of the traditional water deprivation protocol.

Tolerance and osmotic response to food deprivation and salt loading in the herbivorous non-drinking Moroccan Spiny-tailed lizard Uromastyx nigriventris (Sauria: Agamidae)

Animals inhabiting arid environments use a variety of behavioural and physiological strategies to balance their water and salt budgets. We studied the effects of dehydration and salt loading on osmoregulatory capacities in a large herbivorous desert lizard, the Moroccan Spiny-tailed lizard Uromastyx nigriventris, the family Agamidae. These lizards select plants with a high K+ to Na+ ratio of 15 to 20, and like other herbivorous lizards, effectively eliminate the extra electrolyte load, mainly via a pair of active nasal salt glands, which exude the extra ions from blood. Here we present results of a series of laboratory experiments, which tested a five-week food and water deprivation and the excretory response of nasal salt glands, during a short period of five days, following salt loading by two separated injections of KCl or NaCl at a 5-day interval (4th and 9th days). During food-water deprivation, hypohydrated lizards lost 32% of their initial body mass with a substantial decrease of their Body Condition Index and the tail volume as an index of energy (fat and then potential metabolic water) storage. Plasma osmolality significantly increased by 20%. There were also significantly increased plasma sodium, chloride, and total protein concentrations. On the other hand, there was no significant decrease in the plasma glucose level. Most of the salt loaded lizards secreted far more K+ than Na+ via the nasal glands, even after NaCl loading. The K+/Na+ ratio decreased only after two to three repetitive NaCl injections but insufficient Na+ was eliminated. Two successive KCl injections were successfully eliminated, but daily natural average K+ administration induced progressive hyperkaliemia. These experimental data agreed with previous observations showing variations of plasma Na+ and K+ concentrations in free-living lizards. The nasal gland constitutes the main route of Cl− excretion but the Cl−/(Na+ + K+) ratio may vary according to observations in other herbivorous species.

PSIX-25 Feed and water deprivation at weaning and subsequent heat stress impacts serum and ileal markers of gastrointestinal integrity in nursery pigs

Social and environmental stressors impact nursery pig performance and may be linked to gastrointestinal dysfunction. The current experiment was conducted to investigate the impact of feed and water deprivation on serum stress markers and ileal mucosal gene expression in nursery pigs. Mixed-sex pigs were allotted on the basis of IBW (7.0 kg ± 0.89) in a RCBD with treatments in a split-plot arrangement and consisting of the whole-plot factor of with or without a 24-h feed and water deprivation at weaning and the sub-plot factor of with or without a cyclic 3-d heat stress starting 27 d post-weaning. On 1, 27, and 30 d post-weaning one pig from each pen was selected, blood was collected for measurement of serum cortisol, corticotrophin releasing factor (CRF), and endotoxins, and an ileal mucosal scraping was taken and gene expression of claudin 1 (CL-1), occludin (OC), and zonula occludens 1 (ZO-1) were measured by RT PCR. There was an increase (P < 0.05) in serum CRF and endotoxins and a tendency for an increase (P = 0.09) in serum cortisol due to the deprivation 1 d post-weaning. Further, there was a tendency for an increase (P < 0.10) in serum endotoxins and CRF due to the deprivation at 27 and 30 d post-weaning, respectively. Gene expression of CL-1 tended to increase (P = 0.10), and OC decreased (P = 0.05) due to the deprivation 1 d post-weaning. Expression of the OC gene decreased (P < 0.05) due to the deprivation 27 d post-weaning and OC and ZO-1 gene expression tended to decrease (P = 0.07) due to the heat stress 30 d post-weaning. These results show that post-weaning stress events alter serum stress markers and impact intestinal barrier function.

Direct and Indirect Transcriptional Effects of Abiotic Stress in Zea mays Plants Defective in RNA-Directed DNA Methylation

Plants respond to abiotic stress stimuli, such as water deprivation, through a hierarchical cascade that includes detection and signaling to mediate transcriptional and physiological changes. The phytohormone abscisic acid (ABA) is well-characterized for its regulatory role in these processes in response to specific environmental cues. ABA-mediated changes in gene expression have been demonstrated to be temporally-dependent, however, the genome-wide timing of these responses are not well-characterized in the agronomically important crop plant Zea mays (maize). ABA-mediated responses are synergistic with other regulatory mechanisms, including the plant-specific RNA-directed DNA methylation (RdDM) epigenetic pathway. Our prior work demonstrated that after relatively long-term ABA induction (8 h), maize plants homozygous for the mop1-1 mutation, defective in a component of the RdDM pathway, exhibit enhanced transcriptional sensitivity to the phytohormone. At this time-point, many hierarchically positioned transcription factors are differentially expressed resulting in primary (direct) and secondary (indirect) transcriptional outcomes. To identify more immediate and direct MOP1-dependent responses to ABA, we conducted a transcriptomic analysis using mop1-1 mutant and wild type plants treated with ABA for 1 h. One h of ABA treatment was sufficient to induce unique categories of differentially expressed genes (DEGs) in mop1-1. A comparative analysis between the two time-points revealed that distinct epigenetically-regulated changes in gene expression occur within the early stages of ABA induction, and that these changes are predicted to influence less immediate, indirect transcriptional responses. Homology with MOP1-dependent siRNAs and a gene regulatory network (GRN) were used to identify putative immediate and indirect targets, respectively. By manipulating two key regulatory networks in a temporal dependent manner, we identified genes and biological processes regulated by RdDM and ABA-mediated stress responses. Consistent with mis-regulation of gene expression, mop1-1 homozygous plants are compromised in their ability to recover from water deprivation. Collectively, these results indicate transcriptionally and physiologically relevant roles for MOP1-mediated regulation of gene expression of plant responses to environmental stress.

The usefulness of copeptin for the diagnosis of nephrogenic diabetes insipidus in infancy: a case report

Objectives We report a case of an infant with nephrogenic diabetes insipidus (NDI) diagnosed by the measurement of serum copeptin. There is only one study that previously evaluated the use of copeptin measurement in a pediatric patient. Case presentation We present a 10-month-old child with polyuria–polydipsia syndrome (PPS) and hypernatremia that could not support water restriction due to increased risk of dehydration and worsening of his condition. Therefore, plasma measurement of copeptin allowed the diagnosis of NDI. Conclusions The water deprivation test (WDT) is considered the gold standard for diagnosis in PPS. However, WDT has serious limitations regarding its interpretation. Furthermore, the WDT can cause dehydration and hypernatremia, especially in young children. Therefore, the measurement of plasma copeptin seems to be a promising method to perform an earlier, safer, and accurate investigation of PPS. Up to now, our study is the second to report the usefulness of copeptin in children.