Subsequent Recovery Period Research Articles

Effects of Atrazine and Chlorpyrifos on Autophagy-Related Genes in the Brain of Common Carp: Health-Risk Assessments.

This study assessed the impacts of atrazine (ATR), chlorpyrifos (CPF), and a combined ATR/CPF exposure on the brain of common carp (Cyprinus carpio L.). The carp were sampled after a 40-days exposure to CPF and ATR, individually or in combination, followed by a 40-days recovery period to measure autophagy and antioxidant activity. The results indicate that the anti-superoxide anion and anti-hydroxy radical activities decreased upon exposure to ATR, CPF, and the ATR/CPF combination but increased after a subsequent 40-days recovery period. Quantitative real-time PCR and Western blot analyses revealed that the mRNA and protein levels of LC3B and dynein in common carp decreased significantly after exposure to ATR and CPF alone or in combination. Moreover, the mRNA and protein levels of beclin1 gene decreased significantly only in the 116 and 11.3μg/L treatment groups. However, the mRNA and protein levels of all tested genes increased significantly after a 40-days recovery. Transmission electron microscope demonstrated the occurrence of autolysosomes in the recovery groups but not in the exposure groups. These results suggest that exposure to ATR, CPF, or their combination promotes oxidative stress and autophagic responses in the brain of common carp.

The Effect of Continuous Positive Airway Pressure in a Mouse Model of Hyperoxic Neonatal Lung Injury

Background: Continuous positive airway pressure (CPAP) and supplemental oxygen have become the mainstay of neonatal respiratory support in preterm infants. Although oxygen therapy is associated with respiratory morbidities including bronchopulmonary dysplasia (BPD), the long-term effects of CPAP on lung function are largely unknown. We used a hyperoxia-induced mouse model of BPD to explore the effects of daily CPAP in the first week of life on later respiratory system mechanics. Objective: We wanted to test the hypothesis that daily CPAP in a newborn-mouse model of BPD improves longer-term respiratory mechanics. Methods: Mouse pups from C57BL/6 pregnant dams were exposed to room air (RA) or hyperoxia (50% O₂, 24 h/day) for the first postnatal week with or without exposure to daily CPAP (6 cm H₂O, 3 h/day). Respiratory system resistance (Rrs) and compliance (Crs) were measured following a subsequent 2-week period of RA recovery. Additional measurements included radial alveolar and macrophage counts. Results: Mice exposed to hyperoxia had significantly elevated Rrs, decreased Crs, reduced alveolarization and increased macrophage counts at 3 weeks when compared to RA-treated mice. Daily CPAP treatment significantly improved Rrs, Crs and alveolarization and decreased lung macrophage infiltration in the hyperoxia-exposed pups. Conclusions: We have demonstrated that daily CPAP had a longer-term benefit on baseline respiratory system mechanics in a neonatal mouse model of BPD. We speculate that this beneficial effect of CPAP was the consequence of a decrease in the inflammatory response and resultant alveolar injury associated with hyperoxic lung injury in newborns.

Diffusion of di(2-ethylhexyl)phthalate in PVC quantified by ATR-IR spectroscopy

Di(2-ethylhexyl) phthalate (DEHP) is a commonly employed plasticizer for poly(vinyl chloride) (PVC). Diffusion of di(2-ethylhexyl) phthalate (DEHP) takes place when plasticized PVC is immersed in hexane. Rather than determining DEHP concentration in the immersion solvent, an IR window for PVC from 1500 to 1900 cm−1 enabled direct measurement of HADD (hexane assisted DEHP diffusion) using ATR-IR by monitoring the DEHP carbonyl absorption at 1715 cm−1. The present study employed PVC containing 45% plasticizer (PVC-45, tygon tubing). A fast diffusion process was observed for short times (<30 s) while a slower diffusion process occurred at longer immersion times (30 s < t < 6 min). A synergistic increase in mesosurface Tg toward that of PVC coupled with low solubility of hexane in PVC accounts for decreased DEHP diffusion rates (10−12–10−13 cm2 s−1) for immersion times > 30 s. Analysis by XPS shows the outermost surface is PVC free of plasticizer after 6 min immersion. After an induction period, back diffusion coefficients Db were estimated from peak areas during a subsequent fast recovery period (10−14–10−15 cm2 sec−1). A model is proposed for the induction period associated with re-plasticization that involves a change from a mesosurface glass to a gel like solid.

Assessing the influence of mechanical ventilation on blood gases and blood pressure in rattlesnakes

ObjectiveTo characterize the impact of mechanical positive pressure ventilation on heart rate (HR), arterial blood pressure, blood gases, lactate, glucose, sodium, potassium and calcium concentrations in rattlesnakes during anesthesia and the subsequent recovery period. Study designProspective, randomized trial. AnimalsTwenty one fasted adult South American rattlesnakes (Crotalus durissus terrificus). MethodsSnakes were anesthetized with propofol (15 mg kg−1) intravenously, endotracheally intubated and assigned to one of four ventilation regimens: Spontaneous ventilation, or mechanical ventilation at a tidal volume of 30 mL kg−1 at 1 breath every 90 seconds, 5 breaths minute−1, or 15 breaths minute−1. Arterial blood was collected from indwelling catheters at 30, 40, and 60 minutes and 2, 6, and 24 hours following induction of anesthesia and analyzed for pH, PaO2, PaCO2, and selected variables. Mean arterial blood pressure (MAP) and HR were recorded at 30, 40, 60 minutes and 24 hours. ResultsSpontaneous ventilation and 1 breath every 90 seconds resulted in a mild hypercapnia (PaCO2 22.4 ± 4.3 mmHg [3.0 ± 0.6 kPa] and 24.5 ± 1.6 mmHg [3.3 ± 0.2 kPa], respectively), 5 breaths minute−1 resulted in normocapnia (14.2 ± 2.7 mmHg [1.9 ± 0.4 kPa]), while 15 breaths minute−1 caused marked hypocapnia (8.2 ± 2.5 mmHg [1.1 ± 0.3 kPa]). Following recovery, blood gases of the four groups were similar from 2 hours. Anesthesia, independent of ventilation was associated with significantly elevated glucose, lactate and potassium concentrations compared to values at 24 hours (p < 0.0001). MAP increased significantly with increasing ventilation frequency (p < 0.001). HR did not vary among regimens. Conclusions and clinical relevanceMechanical ventilation had a profound impact on blood gases and blood pressure. The results support the use of mechanical ventilation with a frequency of 1–2 breaths minute−1 at a tidal volume of 30 mL kg−1 during anesthesia in fasted snakes.

The synergistic potential of the azole fungicides prochloraz and propiconazole toward a short α-cypermethrin pulse increases over time in Daphnia magna

Pyrethroid insecticides are highly toxic to non-target aquatic invertebrates. Their high toxicity is synergized when co-occurring with azole fungicides in the aquatic environment. Little is known about the importance of synergy, when pyrethroids only occur during a short pulse of a few hours, as it is likely to happen in the environment, nor about the persistence of synergy over time. This study analyzed the synergistic potential of the fungicides propiconazole and prochloraz toward Daphnia magna, when exposed to a pulse (7.2h) of α-cypermethrin at different concentrations (average pulse concentrations 0.07–11nM). Immobilization was monitored during exposure and a subsequent recovery period (87.5h) with and without continuous co-exposure to the azoles (1.4 and 1.7μM, respectively). EC50 values for immobilization decreased exponentially over time with a higher rate in the presence of the azoles. EC50 values for α-cypermethrin determined at the end of the experiment were 3.3±0.5nM in the absence of azoles and 0.26±0.04, and 0.08±0.01nM in the presence of propiconazole and prochloraz, respectively. The synergistic potential of the azoles was strongly dependent on time: no synergism could be detected during the pulse, but with azole co-exposure EC50 values decreased during the recovery period by a factor of up to 13 (propiconazole) and 61 (prochloraz) compared to values without azole exposure. Such high synergistic ratios have not been reported for pesticide mixtures in literature before. Our findings highlight that a pulse of the pyrethroid α-cypermethrin is synergized far beyond the actual pulse and beyond standardized test durations. Long post-exposure times are therefore mandatory in order to capture full synergism.

Team adaptation to an unforeseen system failure: Limits of the potential aids of shared knowledge and standardized communication

The present study investigates the potential benefits of a team’s shared knowledge and standardized communication in adapting to an unforeseen change by combining literature on adaptation and team performance. Each of 20 teams performed a dynamic team task and was suddenly confronted with a simulated partial system breakdown. Results show that a methodological framework designed to describe performance adaptation to an unforeseen change in individuals can also be used to model performance adaptation in teams. The system failure was followed by a performance drop and a subsequent period of gradual performance recovery. Accuracy of teams’ shared knowledge correlated positively with performance before and after the change, confirming and extending the literature on shared mental models. However, the amount of knowledge similarity did not aid teams in adapting to the unforeseen system breakdown. In addition, improving teams’ standardized communication had no damping effect on the sudden performance drop and neither helped them during the subsequent recovery period. These results show that even though shared knowledge and efficient communication are of high value to team performance and success, these characteristics are limited in aiding adaptive team performance after unforeseen unique changes that force team members to update their strategies.

Immediate and residual effects of heat stress and restricted intake on milk protein and casein composition and energy metabolism

The effects of heat stress on dairy production can be separated into 2 distinct causes: those effects that are mediated by the reduced voluntary feed intake associated with heat stress, and the direct physiological and metabolic effects of heat stress. To distinguish between these, and identify their effect on milk protein and casein concentration, mid-lactation Holstein-Friesian cows (n=24) were housed in temperature-controlled chambers and either subjected to heat stress [HS; temperature-humidity index (THI) ~78] or kept in a THI<70 environment and pair-fed with heat-stressed cows (TN-R) for 7 d. A control group of cows was kept in a THI<70 environment with ad libitum feeding (TN-AL). A subsequent recovery period (7 d), with THI<70 and ad libitum feeding followed. Intake accounted for only part of the effects of heat stress. Heat stress reduced the milk protein concentration, casein number, and casein concentration and increased the urea concentration in milk beyond the effects of restriction of intake. Under HS, the proportion in total casein of αS1-casein increased and the proportion of αS2-casein decreased. Because no effect of HS on milk fat or lactose concentration was found, these effects appeared to be the result of specific downregulation of mammary protein synthesis, and not a general reduction in mammary activity. No residual effects were found of HS or TN-R on milk production or composition after THI<70 and ad libitum intake were restored. Heat-stressed cows had elevated blood concentrations of urea and Ca, compared with TN-R and TN-AL. Cows in TN-R had higher serum nonesterified fatty acid concentrations than cows in HS. It was proposed that HS and TN-R cows may mobilize different tissues as endogenous sources of energy.

Genome-wide transcriptional responses to carbon starvation in nongrowing Lactococcus lactis.

This paper describes the transcriptional adaptations of nongrowing, retentostat cultures of Lactococcus lactis to starvation. Near-zero-growth cultures (μ = 0.0001 h(-1)) obtained by extended retentostat cultivation were exposed to starvation by termination of the medium supply for 24 h, followed by a recovery period of another 24 h by reinitiating the medium supply to the retentostat culture. During starvation, the viability of the culture was largely retained, and the expression of genes involved in transcription and translational machineries, cell division, and cell membrane energy metabolism was strongly repressed. Expression of these genes was largely recovered following the reinitiation of the medium supply. Starvation triggered the elevated expression of genes associated with synthesis of branched-chain amino acids, histidine, purine, and riboflavin. The expression of these biosynthesis genes was found to remain at an elevated level after reinitiation of the medium supply. In addition, starvation induced the complete gene set predicted to be involved in natural competence in L. lactis KF147, and the elevated expression of these genes was sustained during the subsequent recovery period, but our attempts to experimentally demonstrate natural transformation in these cells failed. Mining the starvation response gene set identified a conserved cis-acting element that resembles the lactococcal CodY motif in the upstream regions of genes associated with transcription and translational machineries, purine biosynthesis, and natural transformation in L. lactis, suggesting a role for CodY in the observed transcriptome adaptations to starvation in nongrowing cells.

Characterising multi-level effects of acute pressure exposure on a shallow-water invertebrate: insights into the kinetics and hierarchy of the stress response.

Hydrostatic pressure is an important, ubiquitous, environmental variable of particular relevance in the marine environment. However, it is widely overlooked despite recent evidence that some marine ectotherms may be demonstrating climate-driven bathymetric range shifts. Wide-ranging effects of increased hydrostatic pressure have been observed from the molecular through to the behavioural level. Still, no study has simultaneously examined these multiple levels of organisation in a single experiment in order to understand the kinetics, hierarchy and interconnected nature of such responses during an acute exposure, and over a subsequent recovery period. Here, we quantify the transcription of a set of previously characterised genes during and after acute pressure exposure in adults of the shrimp Palaemonetes varians. Further, we perform respiratory rate and behavioural analysis over the same period. Increases in expression of genes associated with stress and metabolism were observed during and after high-pressure exposure. Respiratory rate increased during exposure and into the recovery period. Finally, differential behaviour was observed under elevated hydrostatic pressure in comparison to ambient pressure. Characterising generalised responses to acute elevated pressure is a vital precursor to longer-term, acclimation-based pressure studies. Results provide a novel insight into what we term the overall stress response (OSR) to elevated pressure; a concept that we suggest to be applicable to other environmental stressors. We highlight the importance of considering more than a single component of the stress response in physiological studies, particularly in an era where environmental multi-stressor studies are proliferating.

Paradoxical (REM) sleep deprivation in mice using the small-platforms-over-water method: polysomnographic analyses and melanin-concentrating hormone and hypocretin/orexin neuronal activation before, during and after deprivation.

Studying paradoxical sleep homeostasis requires the specific and efficient deprivation of paradoxical sleep and the evaluation of the subsequent recovery period. With this aim, the small-platforms-over-water technique has been used extensively in rats, but only rare studies were conducted in mice, with no sleep data reported during deprivation. Mice are used increasingly with the emergence of transgenic mice and technologies such as optogenetics, raising the need for a reliable method to manipulate paradoxical sleep. To fulfil this need, we refined this deprivation method and analysed vigilance states thoroughly during the entire protocol. We also studied activation of hypocretin/orexin and melanin-concentrating hormone neurones using Fos immunohistochemistry to verify whether mechanisms regulating paradoxical sleep in mice are similar to those in rats. We showed that 48 h of deprivation was highly efficient, with a residual amount of paradoxical sleep of only 2.2%. Slow wave sleep and wake quantities were similar to baseline, except during the first 4 h of deprivation, where slow wave sleep was strongly reduced. After deprivation, we observed a 124% increase in paradoxical sleep quantities during the first hour of rebound. In addition, 34% of hypocretin/orexin neurones were activated during deprivation, whereas melanin-concentrated hormone neurones were activated only during paradoxical sleep rebound. Corticosterone level showed a twofold increase after deprivation and returned to baseline level after 4 h of recovery. In summary, a fairly selective deprivation and a significant rebound of paradoxical sleep can be obtained in mice using the small-platforms-over-water method. As in rats, rebound is accompanied by a selective activation of melanin-concentrating hormone neurones.

Therapeutic application of electrical stimulation and constraint induced movement therapy in perinatal brachial plexus injury: A case report

Infants and children with perinatal brachial plexus injury (PBPI) have motion limitations in the shoulder, elbow, forearm and hand that are dependent on the level of injury and degree of recovery. The injury and subsequent recovery period occur during critical periods of central and spinal neural development placing infants and children at-risk for developmental disregard and disuse of the affected arm and hand. A case report outlines the therapy and surgical interventions provided in the first 2 years of life for a child with global PBPI and a positive Horner's sign. Electrical stimulation and constraint induced movement therapy provided sequentially were effective therapy interventions. Neurosurgery to repair the brachial plexus was performed at an optimal time period.2 The Assisting Hand Assessment,12 Modified Mallet13 and Active Movement Scale14 are effective outcome measures in PBPI and served as valuable guides for therapy intervention.Oxford Level of Evidence: 3b; Individual Case Control Study.

Protecting the petroleum industry: renewed government aid to fossil fuel producers

The dual pressures of the global economic crisis in 2008 and high crude prices through the subsequent recovery period have prompted oil-producing countries to adopt a wide range of protectionist measures including subsidies in all forms and trade and investment restrictions. Focusing on fiscal and industrial policy adjustments in the UK and the People's Republic of China since 2008, this paper argues that both governments have sought an increase in tax contributions from the corporate sector in exchange for intensified, targeted support for specific capital investments that will address the challenges of overall decline in domestic oil production and new field exploration and oil recovery opportunities. These novel “rent-sharing” schemes – inadequately captured in recent academic debates over precise measurements of fuel subsidies – raise concerns for fair competition in the upstream market and politicians’ long-term commitment to the transitioning of energy mix toward green and renewable sources.

Effect of long-term sleep restriction and subsequent recovery sleep on the diurnal rhythms of white blood cell subpopulations.

While acute modifications of sleep duration induces a wide array of immune function alterations, less is known of how longer periods with insufficient sleep affect immune functions and how they return to normal once recovery sleep is obtained. The purpose of the present study was to investigate the effects of five days of restricted sleep and a subsequent 7-day period of sleep recovery on white blood cell (WBC) subpopulation count and diurnal rhythms. Nine healthy males participated in a sleep protocol consisting of two baseline days (8h of sleep/night), five nights with restricted sleep (4h of sleep/night) and seven days of recovery sleep (8h of sleep/night). During nine of these days, blood was drawn hourly during night-time end every third hour during daytime, and differential WBC count was analyzed. Gradual increase across the days of sleep restriction was observed for total WBC (p<.001), monocytes (p<.001), neutrophils (p<.001) and lymphocytes (p<.05). Subsequent recovery sleep resulted in a gradual decrease in monocytes (p<.001) and lymphocytes (p=.001), but not in neutrophils that remained elevated over baseline level at the end of the 7-day recovery period. These effects were associated with altered diurnal rhythms of total WBC and neutrophils, restricted sleep being associated with higher levels during the night and at awakening, resulting in a flattening of the rhythm. The diurnal alterations were reversed when recovery sleep was allowed, although the amplitude of total WBC, neutrophils and monocytes was increased at the end of the recovery period in comparison to baseline. Altogether, these data show that long-term sleep restriction leads to a gradual increase of circulating WBC subpopulations and alterations of the respective diurnal rhythms. Although some of the effects caused by five days of restricted sleep were restored within the first days of recovery, some parameters were not back to baseline even after a period of seven recovery days.

Outlining microbial community dynamics during temperature drop and subsequent recovery period in anaerobic co-digestion systems

To improve the stability of anaerobic reactors, more knowledge is required about how the different communities react against operating perturbations and which specific ones respond better. The objective of this work was to monitor the changes in microbial community structure of an anaerobic digester during a temperature drop by applying different complementary molecular techniques. Temperature decrease led to an increase of Bacteroidales order, Porphyromonadaceae family and Bacteroides genus and a decrease in Syntrophomonas and Clostridium genera. Once the temperature was restored, the reactor recovered the steady state performance without requiring any modification in operational conditions or in the microbiome. During the recovery period, Sedimentibacter genus and Porphyromonadaceae family played an important role in the degradation of the accumulated volatile fatty acids. The hydrogenotrophic methanogens appeared to be the keystone archaeal population at low temperatures as well as in the recovery period. This study stands out that the understanding of microbial community dynamics during temperature drop could be utilized to develop strategies for the mitigation of temperature change consequences and speed up the recovery of stable reactor performance.

Effects of leptin on sperm count and morphology in Sprague-Dawley rats and their reversibility following a 6-week recovery period

Altered epididymal sperm count and morphology following leptin treatment has been reported recently. This study examined the effects of 42 days of leptin treatment on sperm count and morphology and their reversibility during a subsequent 56-day recovery period. Twelve-week-old male Sprague-Dawley rats were randomised into four leptin and four saline-treated control groups (n = 6). Intraperitoneal injections of leptin were given daily (60 μg Kg(-1) body weight) for 42 days. Controls received 0.1 ml of 0.9% saline. Leptin-treated animals and their respective age-matched controls were euthanised on either day 1, 21, 42 or 56 of recovery for collection of epididymal spermatozoa. Sperm concentration was determined using a Makler counting chamber. Spermatozoa were analysed for 8-hydroxy-2-deoxyguanosine and DNA fragmentation (Comet assay). Data were analysed using anova. Sperm concentration was significantly lower but fraction of abnormal spermatozoa, and levels of 8-hydroxy-2-deoxyguanosine were significantly higher in leptin-treated rats on day 1 of recovery. Comet assays revealed significant DNA fragmentation in leptin-treated rats. These differences were reduced by day 56 of recovery. It appears that 42 days of leptin treatment to Sprague-Dawley rats has significant adverse effects on sperm count and morphology that reverse following discontinuation of leptin treatment.

Prochloraz causes irreversible masculinization of zebrafish (Danio rerio).

The aim of the present study was to investigate the persistence of endocrine effects by prochloraz, a fungicide known to have multiple effects on the endocrine system of vertebrates. Since discontinuous exposure is particularly relevant in aquatic ecosystems, an exposure scenario with an exposure phase and a subsequent recovery period was chosen to assess the potential for reversibility of effects by prochloraz on the sexual development of zebrafish (Danio rerio). Zebrafish were exposed to different concentrations of prochloraz (10-300 μg/L) until 60 days post hatch (dph), which includes the period of sexual differentiation. For the subsequent 40 days, fish were either held in clean water for depuration or under further continuous exposure. Histological investigations of the gonads revealed persistent effects on sexual differentiation. The sex ratio was skewed towards males and significantly more intersex individuals were found after exposure to prochloraz at 60 dph. No intersex fish, but masculinized sex ratios were still present after the depuration period, documenting that prochloraz irreversibly affects the sexual development of zebrafish.

Effects of light interruption on sleep and viability of Drosophila melanogaster.

Light is a very important regulator of the daily sleep rhythm. Here, we investigate the influence of nocturnal light stimulation on Drosophila sleep. Results showed that total daytime sleep was reduced due to a decrease in daytime sleep episode duration caused by discontinuous light stimulation, but sleep was not strongly impacted at nighttime although the discontinuous light stimulation occurred during the scotophase. During a subsequent recovery period without light interruption, the sleep quality of nighttime sleep was improved and of daytime sleep reduced, indicating flies have a persistent response to nocturnal light stimulation. Further studies showed that the discontinuous light stimulation damped the daily rhythm of a circadian light-sensitive protein cryptochrome both at the mRNA and protein levels, which subsequently caused disappearance of circadian rhythm of the core oscillator timeless and decrease of TIMLESS protein at nighttime. These data indicate that the nocturnal light interruption plays an important role in sleep through core proteins CRYTOCHROME and TIMLESS, Moreover, interruption of sleep further impacted reproduction and viability.

Effects of chemical stress and food limitation on the energy reserves and growth of turbot, Scophthalmus maximus.

The objective of the present study is to examine the growth and energetic performance of juvenile turbot after exposure to contaminated sediment and during the subsequent recovery period with or without food limitation. We designed a two-step experiment by first exposing juvenile turbot to harbour sediment for 26 days and then transferring them to clean sea water with different frequencies of feeding for 35 days. Without food limitation, fish previously exposed to contaminated sediment compensated for weight, length and lipid reserve losses; we did not record any differences in size, Fulton's K condition index and triacylglycerol/sterol (TAG/ST) ratio after the 35-day depuration period compared to the reference fish. This result could be related to the compensatory growth mechanism observed in a wide range of fish species following a period of growth depression. With food limitation during the 35-day depuration period, recovery growth was not sufficient to restore length and weight values similar to the reference fish. Moreover, turbot previously exposed to contaminated sediment and subsequently fed twice or once a week exhibited extremely low TAG/ST ratios, but the reference fish submitted to the same restrictive feeding conditions did not. This study indicates that juvenile fish affected by chemical pollution can improve their biological performance if pollution events are followed by a period of abundant food. However, if pollution events occur during periods of food scarcity, e.g. in winter, storage of energy reserves will be compromised.

The Impact of Playing Strategies on Recovery in Low‐Goal Polo‐Ponies

IntroductionPolo is a popular sport; teams compete in matches comprising 4–6 chukkas, each between 7–10 minutes. Players commonly change ponies between chukkas, but changing within a chukka, known as ‘half‐chukkering’ is permitted. This is believed to ‘save’ the horse by preventing fatigue and facilitating recovery. This study evaluated the impact of half versus full chukka playing strategies in low goal polo; team handicap 0–4 goals.MethodsResting heart rates (HR) were established for 48 polo‐ponies deemed of comparable fitness by riders, competing in low‐goal tournaments; riders selected their playing strategy either full‐chukka (n = 32) or half‐chukkering (n = 16). HR was recorded at the end of play and during recovery; for the full‐chukka and 2nd half‐chukka period at +2, 4, 6, 10 and 20 minutes, and until ponies returned to play after the 1st half‐chukka, &gt;6 minutes. ANOVA analyses investigated HR variation during recovery between ponies playing full‐chukkas and the 2nd half‐chukka period, and within the two half‐chukkas played.ResultsAt the end of play, full‐chukka horses′ mean HR were 95 ± 13.1 bpm, compared to their 2nd half‐chukka counterparts at 103 ± 7.7 bpm, which was not significant. Significant differences in HR did occur throughout the subsequent recovery period: at 2 minutes (P&lt;0.003) and between 4–20 minutes (P&lt;0.0001), with full‐chukka ponies recovering faster. No differences occurred during recovery between the two half‐chukka periods although mean HR differed at the end of play (1st: 100 ± 7.7 bpm; 2nd: 103 ± 7.8 bpm).ConclusionsPlaying strategies influence cardiovascular recovery; full‐chukka ponies recover faster than those playing in consecutive half‐chukkas. Preparation of half‐chukka ponies may predispose to reduced fitness levels compared to full‐chukka ponies. Implementing a full‐chukka playing strategy is advocated to optimise pony welfare and performance in low‐goal polo.Ethical Animal ResearchEthical approval for the study was granted from the UWE Hartpury Ethics Committee. Explicit owner informed consent for participation in this study is not stated. Sources of funding: none. Competing interests: none.

Environmental cadmium exposure impacts physiological responses in Manila clams.

The physiological responses of marine bivalves to chronic cadmium (Cd) exposure at sub-lethal concentrations have been well documented. As of now, few studies have examined the effect of Cd exposure and subsequent recovery period at environmentally realistic concentrations. In this study, environmentally, Cd exposures were performed to assess the physiological responses of the Manila clam Ruditapes philippinarum. The clams were exposed to waterborne Cd at two environmentally realistic concentrations (4 and 40μgL(-1)) for 35days and then allowed to recover for another 35days. The accumulation and elimination of Cd in R. philippinarum were tissue-specific and dose- and time-dependent. Cd accumulation increased sharply in the digestive gland, and Cd elimination was rapid in the gill. Major physiological responses, including clearance rate, absorption efficiency, respiration rate, excretion rate, oxygen to nitrogen ratio, and scope for growth, were significantly affected by Cd exposure. Yet, the clams exposed to 4-μgL(-1) Cd were able to quickly recover their normal physiological processes and clearly exhibited catch-up growth once they were transferred to clean seawater. Hence, R. philippinarum can exhibit good physiological plasticity when confronted with moderately environmental Cd exposure. All physiological responses measured exhibited a highly significant and generally predictable correlation with tissue Cd concentration, which in turn, reflected environmentally realistic exposure conditions. Our results further confirm that the measurement of physiological responses is a sensitive method for assessing stress at environmentally realistic metal concentrations.