The gene expression response of the catadromous perciform barramundi Lates calcarifer to an acute heat stress

The acute heat-shock response of the tropical estuarine fish species barramundi Lates calcarifer as indicated by the expression of genes within stress (hsp 90AA, hsp 90AB, hsp 70 and hsc 70), metabolic (cisy, cco II and ldh) and growth (igf1 and mstn 1) related pathways was examined following an increase in water temperature from 28 to 36° C over 30 min. Lates calcarifer were maintained at the acute stress temperature of 36° C for 1 h before being returned to 28° C and allowed to recover at this temperature for a further 2 weeks. Muscle tissue sampling over the experimental period allowed for the expression quantification of stress, metabolic and growth-related genes via quantitative real-time polymerase chain reaction (qrt-PCR) where a robust and reliable normalization approach identified both α-tub and Rpl8 as appropriate genes for the analysis of gene expression in response to an acute heat stress. hsp90AA and hsp70 of the inducible heat-shock response pathway showed a massive up-regulation of gene expression in response to heat stress, whilst the constitutive heat-shock genes hsp90AB and hsp70 showed no change over the course of the experiment and a small increase after 2 weeks of recovery, respectively. Of the three genes representing the metabolic pathway (cisy, cco II and ldh) only cco II changed significantly showing a decrease in gene expression, which may suggest a small suppression of aerobic metabolism. igf1 of the growth pathway showed no significant differences in response to an acute heat stress, whilst mstn1 increased at the beginning of the heat stress but returned to basal levels soon after. Overall, the results demonstrate that an acute heat stress in L. calcarifer caused a significant increase in the expression of genes from the stress response pathway and a possible decrease in aerobic metabolism with only relatively minor changes to the growth pathway highlighting the hardy nature of L. calcarifer and its resilience in coping with sudden temperature changes routinely encountered within its natural environment.     

Welfare implication of measuring heart rate and heart rate variability in dairy cattle: literature review and conclusions for future research

Heart rate (HR) measurements have been used to determine stress in livestock species since the beginning of the 1970s. However, according to the latest studies in veterinary and behaviour–physiological sciences, heart rate variability (HRV) proved to be more precise for studying the activity of the autonomic nervous system. In dairy cattle, HR and HRV indices have been used to detect stress caused by routine management practices, pain or milking. This review provides the significance of HR and HRV measurements in dairy cattle by summarising current knowledge and research results in this area. First, the biological background and the interrelation of the autonomic regulation of cardiovascular function, stress, HR and HRV are discussed. Equipment and methodological approaches developed to measure interbeat intervals and estimate HRV in dairy cattle are described. The methods of HRV analysis in time, frequency and non-linear domains are also explained in detail emphasising their physiological background. Finally, the most important scientific results and potential possibilities for future research are presented.     

Transcriptional profiling of mouse liver in response to chronic heat stress

Mice were used to study the effects of chronic heat stress on hepatic gene expression. Twenty-five mice were allocated to either chronic heat stress (34 °C) or control (24 °C) conditions for a period of 2 weeks from 47 to 60 d of age. Nineteen genes differentially expressed in liver were identified using DNA microarrays. Genes involved in the anti-oxidant pathway and metabolism were up-regulated. Genes involved in generation of reactive oxygen radicals and mitochondrial expressed genes were down-regulated. Enzyme activity measurements confirmed the array results. Mice exposed to chronic heat stress showed signs of increased oxidative stress in liver cells.     

Effect of heat stress and feeding phosphorus levels on pig electron transport chain gene expression

The purpose of this study was to evaluate the effect of temperature and different levels of available phosphorus (aP) on the expression of nine genes encoding electron transport chain proteins in the Longissimus dorsi (LD) muscle of pigs. Two trials were carried out using 48 high-lean growth pigs from two different growth phases: from 15 to 30 kg (phase 1) and from 30 to 60 kg (phase 2). Pigs from growth phase 1 were fed with three different levels of dietary aP (0.107%, 0.321% or 0.535%) and submitted either to a thermoneutral (24°C and RH at 76%) or to a heat stress (34°C and RH at 70%) environment. Pigs from growth phase 2 were fed with three different levels of dietary aP (0.116%, 0.306% or 0.496%) and submitted either to a thermoneutral (22ºC and RH at 77%) or to a heat stress (32ºC and RH at 73%) environment. Heat stress decreased (P<0.001) average daily feed intake at both growth phases. At 24°C, pigs in phase 1 fed the 0.321% aP diet had greater average daily gain and feed conversion (P<0.05) than those fed the 0.107% or 0.535% while, at 34°C pigs fed the 0.535% aP had the best performance (P<0.05). Pigs from phase 2 fed the 0.306% aP had best performance in both thermal environments. Gene expression profile was analyzed by quantitative real-time polymerase chain reaction. Irrespective of growing phase, the expression of six genes was lower (P<0.05) at high temperature than at thermoneutrality. The lower expression of these genes under high temperatures evidences the effects of heat stress by decreasing oxidative metabolism, through adaptive physiological mechanisms in order to reduce heat production. In pigs from phase 1, six genes were differentially expressed across aP levels (P<0.05) in the thermoneutral and one gene in the heat stress. In pigs from phase 2, two genes were differentially expressed across aP levels (P<0.05) in both thermal environments. These data revealed strong evidence that phosphorus and thermal environments are key factors to regulate oxidative phosphorylation with direct implications on animal performance.     

Effects of acute and chronic heat stress on feed sorting behaviour of lactating dairy cows

Nutritional strategies to mitigate the negative effects of heat stress on animal welfare and productivity often involve changes in ration formulation. However, cattle commonly sort their ration in favour of certain components, and it is not clear how feed sorting responds to heat stress. This study investigated the association between heat stress and feed sorting behaviour. Lactating Holstein dairy cows (n = 32; parity = 2.8±1.2; mean±SD) were housed in a free stall barn and milked 3×/day. Cows were fed individually using the Calan Broadbent Feeding System and offered ad libitum access to a total mixed ration (containing on a dry matter basis: 3.3% ryegrass hay, 16.5% ryegrass baleage, 24.7% corn silage, 11.1% brewers grains, 19.7% ground corn, 19.8% concentrate and 4.9% protein/mineral supplement), provided 1×/day. Beginning at 186±60 days in milk, cows were exposed to either: heat stress conditions (HT; n = 15) (average temperature–humidity index: 77.6), or evaporative cooling (CL; n = 17), consisting of misters and fans over the freestall and feed bunks. Data were collected during a 4-day baseline period, and two 4-day experimental periods: starting at 10 days after implementing treatments (defined as acute heat stress for HT cows), and at 62 days after implementing treatments (defined as chronic heat stress for HT cows). Daily feed intake and physiological responses to heat stress (body temperature, respiration rate) were recorded. Samples of fresh and refused feed were collected daily from individual cows for particle size analysis. The particle size separator had three screens (19, 8 and 1.18 mm) and a bottom pan, resulting in 4 fractions (long, medium, short and fine particles). Feed sorting was calculated as the actual intake of each particle size fraction expressed as a percentage of the predicted intake of that fraction. During both heat stress periods, HT cows sorted for long particles more than CL cows (105.0% v. 100.6%; SE = 1.1). During acute heat stress, HT cows sorted to a greater extent than CL cows against medium and short particles, whereas sorting of these fractions did not differ during chronic heat stress. Body temperature and respiration rate were associated across treatments with the extent of sorting for long particles and against short particles during acute heat stress. These results suggest that feed sorting is particularly influenced during acute heat stress, and that sorting for longer particles may increase in heat stress.     

Sedation or anaesthesia decrease the stress response to electroejaculation and improve the quality of the collected semen in goat bucks

Electroejaculation (EE) is stressful and probably painful; thus the administration of anaesthesia is recommended to decrease those negative effects. However, anaesthesia has a direct risk of provoking death, but sedation is less risky than anaesthesia. At the same time, α2-adrenergic agonists may improve semen quality. Therefore, the aim of the present study was to compare the physiological and behavioural responses indicative of stress and possibly pain, and the semen quality in electroejaculated untreated, anaesthetised or sedated goat bucks. Semen was collected from eight bucks using three different procedures in all them (EE in untreated bucks, EE under sedation or EE under general anaesthesia). The number of vocalizations during EE and the behavioural pattern before and after procedures were recorded. Pain visual analogue scale (VAS) score was also determined during EE. Rectal temperature, heart rate, serum cortisol concentration, biochemical and haematological parameters were measured before and after each procedure, and sperm characteristics were determined. Bucks vocalised more often when untreated than sedated or anaesthetised (P<0.02). The pain VAS score was greater when bucks were untreated than sedated or anaesthetised (P<0.002). The rectal temperature, heart rate, total protein, albumin and haemoglobin concentrations were greater when bucks were untreated than anaesthetised or sedated (P<0.02). Serum cortisol increased after EE (P=0.0006), without differences between procedures. The frequency and duration of lying down after EE were greater when bucks were anaesthetised than sedated or untreated (P<0.05), and were also greater when bucks were sedated than untreated (P<0.05). The number of times that the animal tried to stand up after EE was greater when bucks were anaesthetised than sedated or untreated (P<0.02). The sperm mass motility was greater when bucks were anaesthetised or sedated than when they were untreated (P=0.048). When animals were sedated, the ejaculate contained more spermatozoa with functional plasma membrane (P=0.03) and morphologically normal (P=0.05) than when they were untreated. In conclusion, general anaesthesia and sedation decreased the stress and probably the pain response provoked by EE and especially sedation improved the quality of the semen collected.     

Changes in protein expression in testes of L2 strain Taiwan country chickens in response to acute heat stress

Heat stress causes a decrease of fertility in roosters. Yet, the way acute heat stress affects protein expression remains poorly understood. This study investigated differential protein expression in testes of the L2 strain of Taiwan country chickens following acute heat stress. Twelve 45-week-old roosters were allocated into four groups, including control roosters kept at 25 °C, roosters subjected to 38 °C acute heat stress for 4 hours without recovery, with 2 hours of recovery, and with 6 hours of recovery. Testis samples were collected for morphologic assay and protein analysis. Some of the differentially expressed proteins were validated by Western blot and immunohistochemistry. Abnormal and apoptotic spermatogenic cells were observed at 2 hours of recovery after acute heat stress, especially among the spermatocytes. Two-dimensional difference gel electrophoresis revealed that 119 protein spots were differentially expressed in chicken testes following heat stress, and peptide mass fingerprinting revealed that these spots contained 92 distinct proteins. In the heat-stressed samples, the heat shock proteins, chaperonin containing t-complex, and proteasome subunits were downregulated, and glutathione S-transferase, transgelin, and DJ-1 were upregulated. Our results demonstrate that acute heat stress impairs the processes of translation, protein folding, and protein degradation, and thus results in apoptosis and interferes with spermatogenesis. On the other hand, the increased expression of antioxidant enzymes, including glutathione S-transferase and DJ-1, may attenuate heat-induced damage. These findings may have implications for breeding chickens that can tolerate more extreme conditions.     

集胞藻PCC6803中S2P同源蛋白基因sll0862缺失突变株对热胁迫与氧化胁迫的响应

原核生物中S2P参与应答外界环境刺激,然而行光合作用的蓝细菌-集胞藻PCC6803的S2P同源蛋白功能未知。【目的】考察集胞藻PCC6803中S2P同源蛋白sll0862是否参与外界环境刺激的应答。【方法】监测在高温和氧化胁迫的条件下sll0862基因缺失突变株与野生株在生长速率或存活率上的差异,利用水样调制叶绿素荧光仪(water-PAM,脉冲-振幅-调制叶绿素荧光仪)测量在高温和氧化胁迫的条件下突变株与野生株叶绿素荧光参数的差异,来考察其光合作用差异。【结果】sll0862突变株与野生株在正常的培养环境中生长速率并无差异,但是将sll0862突变株与野生株在48℃加热处理半小时后,sll0862突变株的存活率明显低于野生株。当初始OD730值为0.1的藻液中添加终浓度为1 mmol/L双氧水的时候,sll0862突变株的生长速率比野生株明显低,而且氧化胁迫条件下突变株与野生株的调制叶绿素荧光有差异。【结论】集胞藻PCC6803中sll0862基因的缺失导致突变体对高温与氧化胁迫响应出现缺陷,提示有功能的sll0862参与响应热和氧化胁迫。研究结果为进一步阐述S2P同源蛋白sll0862在集胞藻PCC6803中的功能奠定基础。     

Dissecting the proteome dynamics of the early heat stress response leading to plant survival or death in Arabidopsis

In many plant species, an exposure to a sublethal temperature triggers an adaptative response called acclimation. This response involves an extensive molecular reprogramming that allows the plant to further survive to an otherwise lethal increase of temperature. A related response is also launched under an abrupt and lethal heat stress that, in this case, is unable to successfully promote thermotolerance and therefore ends up in plant death. Although these molecular programmes are expected to have common players, the overlapping degree and the specific regulators of each process are currently unknown. We have carried out a high‐throughput comparative proteomics analysis during acclimation and during the early stages of the plant response to a severe heat stress that lead Arabidopsis seedlings either to survival or death. This analysis dissects these responses, unravels the common players and identifies the specific proteins associated with these different fates. Thermotolerance assays of mutants in genes with an uncharacterized role in heat stress demonstrate the relevance of this study to uncover both positive and negative heat regulators and pinpoint a pivotal role of JR1 and BAG6 in heat tolerance.     

Serum proteomics analysis reveals the thermal fitness of crossbred dairy buffalo to chronic heat stress

Chronic heat stress (CHS) reduces the production efficiency of the buffalo dairy industry. Relatively low-abundance proteins with particular functions in biological processes are changed by CHS. The present study aimed to quantify the differences in low-abundance proteins of crossbred dairy buffaloes under CHS and thermal-neutral (TN) conditions. With label-free quantification, 344 low-abundance proteins were identified in serum. Of these, 17 differentially expressed low-abundance proteins with known functions were detected, and six of the differentially expressed proteins related to heat stress were validated with parallel reaction monitoring. Lipase (LPL), glutathione peroxidase 3 (GPX3), cathelicidin-2 (CATHL2), ceruloplasmin (CP), and hemoglobin subunit alpha 1 (HBA1) cooperatively played roles in the thermal fitness of dairy buffalo by decreasing heat production and increasing blood oxygen delivery. Also, dairy buffaloes may adapt to CHS and hypoxia with high levels of RBCs, HBA1 and CP to increase blood oxygen delivery capacity.     

黄连木对干旱胁迫的生理响应

研究了自然干旱条件下黄连木（Pistacia chinensis Bunge）的生理变化。结果表明,随土壤含水量的减少,叶绿素b含量、光合速率、叶片相对含水量与叶水势均下降;叶绿素a和可溶性糖含量、叶绿素a和b的比值及总叶绿素含量呈现上升的趋势;超氧化物歧化酶活性先升后降;丙二醛含量干旱胁迫前期升高,后期变化不明显;净光合速率、气孔导度和蒸腾速率随土壤含水量的减少逐步降低。气孔和可溶性糖含量都是影响黄连木光合速率的关键因子,干旱胁迫前12d光合速率主要受气孔限制,之后为非气孔限制。干旱胁迫前期渗透调节物质以可溶性糖为主,干旱胁迫较重时脯氨酸含量急剧升高,与可溶性糖同时起渗透调节作用。     

Analysis of heart rate variability during exercise stress testing using respiratory information

This paper presents a novel method for the analysis of heart rate variability (HRV) during exercise stress testing enhanced with respiratory information. The instantaneous frequency and power of the low frequency (LF) and high frequency (HF) bands of the HRV are estimated by parametric decomposition of the instantaneous autocorrelation function (ACF) as a sum of damped sinusoids. The instantaneous ACF is first windowed and filtered to reduce the cross terms. The inclusion of respiratory information is proposed at different stages of the analysis, namely, the design of the filter applied to the instantaneous ACF, the parametric decomposition, and the definition of a dynamic HF band. The performance of the method is evaluated on simulated data as well as on a stress testing database. The simulation results show that the inclusion of respiratory information reduces the estimation error of the amplitude of the HF component from 3.5% to 2.4% in mean and related SD from 3.0% to 1.7% when a tuned time smoothing window is used at an SNR of 15 dB. Results from the stress testing database show that information on respiratory frequency produces HF power estimates which closely resemble those from the simulations which exhibited lower SD. The mean SD of these estimates with respect to their mean trends is reduced by 84% (from 0.74×10⁻³ s⁻² to 0.12×10⁻³ s⁻²). The analysis of HRV in the stress testing database reveals a significant decrease in the power of both the LF and HF components around peak stress.     

Association analysis for agronomic traits in wheat under terminal heat stress

Terminal heat stress causes irreversible damage to wheat crop productivity. It reduces the vegetative growth and flowering period that consequently declines the efficiency to capture available stem reserves (carbohydrates) in grains. Markers associated with thermotolerant traits ease in marker assisted selection (MAS) for crop improvement. It identifies the genomic regions associated with thermotolerant traits in wheat, but the scarcity of markers is the major hindrance in crop improvement. Therefore, 158 wheat genotypes were subjected to genotyping with 165 simple sequence repeat markers dispersed on three genomes (A, B and D). Allelic frequency and polymorphic information content values were highest on genome A (5.34 (14% greater than the lowest value at genome D) and 0.715 (3% greater than the lowest value at genome D)), chromosome 4 (5.40 (16% greater than the lowest value at chromosome 2) and 0.725 (5% greater than the lowest value at chromosome 6)) and marker xgwm44 (13.0 (84% greater than the lowest value at marker xbarc148) and 0.916 (46% greater than the lowest value at marker xbarc148)). Bayesian based population structure discriminated the wheat genotypes into seven groups based on genetic similarity indicating their ancestral origin and geographical ecotype. Linkage disequilibrium pattern had highest significant (P < 0.001) linked loci pairs 732 on genome A at r² > 0.1 whereas, 58 on genome B at r² > 0.5. Linkage disequilibrium decay (P < 0.01 and r² > 0.1) had larger LD block (5–10 cM) on genome A. Highly significant MTAs (P < 0.000061) under heat stress conditions were identified for flag leaf area (xwmc336), spikelet per spike (xwmc553), grains per spike (cxfa2147, xwmc418 and xwmc121), biomass (xbarc7) and grain yield (xcfa2147 and xwmc671). The identified markers in this study could facilitate in MAS and gene pyramiding against heat stress in wheat.     

存在睡眠呼吸异常的慢病患者呼吸影响心率变异性的初步分析*

目的: 基于整体整合生理学医学理论提出的呼吸引起循环指标变异的假说,分析研究存在睡眠呼吸异常的慢病患者睡眠期间呼吸和心率变异之间的相关关系。方法: 纳入存在睡眠呼吸异常且呼吸暂停低通气指数（AHI）≥15次/小时的慢病患者11例,签署知情同意书后完成标准化症状限制性极限运动的心肺运动试验（CPET）和睡眠呼吸监测,计算分析病人睡眠期间波浪式呼吸（OB）期与正常平稳呼吸期的呼吸鼻气流、心电图R-R间期心率变异的规律。结果: 存在睡眠呼吸异常的慢病患者CPET峰值摄氧量（Peak VO₂）和无氧阈（AT）为（70.8±13.6）%pred和（71.2±6.1）%pred;CPET有5例存在运动诱发的波浪式呼吸（EIOB）,6例为呼吸不稳定,提示整体功能状态低于正常人。本组慢病患者AHI为每小时（28.8±10.0）次,睡眠呼吸异常总时间占睡眠总时间的比值为（0.38±0.25）;OB周期的平均时间长度为（51.1±14.4）s。本组慢病患者正常平稳呼吸期的呼吸周期数与心率变异周期数的比值（B-n/HRV-B-n）为1.00±0.04,每个呼吸周期节律的心率变异平均幅度（HRV-B-M）为（2.64±1.59） bpm,虽然低于正常人（P<0.05）,但却与无睡眠呼吸异常的慢病患者相似（P>0.05）;HRV-B-M的变异度CV（HRV-B-M的SD/x）为( 0.33±0.11）,期间血氧饱和度（SpO₂）虽略低,但并无明显规律性下降与上升。本组慢病患者的OB期间呼吸周期数与心率变异周期数（OB-B-n/OB-HRV-B-n）比值为（1.22±0.18）,OB期每个呼吸周期节律的心率变异平均幅度（OB-HRV-B-M）为（3.56±1.57）bpm及其变异度（OB-CV =OB-HRV-B-M的SD/x）为（0.59±0.28）,每个OB周期节律的心率变异平均幅度（OB-HRV-OB-M）为（13.75±4.25）bpm,OB期间低通气时SpO₂出现明显的下降,OB期间SpO₂平均变异幅度（OB-SpO₂-OB-M）为（4.79±1.39）%,OB期的OB-B-n/OB-HRV-B-n比值、OB-HRV-OB-M比其正常平稳呼吸期对应指标显著增大（P<0.01）。OB-HRV-B-M虽然与正常平稳呼吸期HRV-B-M相比差异无统计学意义（P>0.05）,但其变异度OB-CV却显著增大（P<0.01）。结论: 睡眠呼吸异常的慢病患者OB期的心率变异幅度大于其正常平稳呼吸期,当呼吸模式发生改变时心率变异也发生明显改变,其平稳呼吸期的呼吸周期数与心率变异周期数的比值与正常人以及无睡眠呼吸异常的慢病患者相同,证实心率变异为呼吸源性;而其OB期间心率变异周期数相对于呼吸周期减少直接源于此时的低通气或者呼吸暂停,心率变异也是呼吸源性。     

热应激环境下蛋鸡肠道微生物菌群多样性

通过分析热应激环境下蛋鸡肠道菌群结构的变化,旨在揭示热应激对肠道微生态环境的影响机理,为探索炎热环境下家禽肠道菌群定植规律提供理论依据。试验选择16周龄济宁百日鸡96只,随机分成对照组[(24±1)℃,Ⅰ]和热应激[(38±1)℃]组,分别在2个人工环境气候舱中饲养,各组设6个重复,每个重复8只,试验持续14 d。采用16S rDNA的变性梯度凝胶电泳(Denatured gradient gel electrophoresis,PCR-DGGE)技术和实时荧光定量(Real-time quantitative RT-PCR)以及多变量统计(Principal Components Analysis,PCA)分析等手段,分析热应激2d(Ⅱ)、7d(Ⅲ)和14 d(Ⅳ)时,对十二指肠、空肠及回肠内容物菌群多样性以及菌群数量变化。PCA分析结果显示,热应激暴露过程中十二指肠部位菌群组成与对照组保持相似的趋势,但热应激2 d时空肠部位菌群组成有分开趋势;到7 d时空肠与回肠部位菌群组成分开明显,而14 d时菌群组成具有明显差异;热应激7、14 d时空肠和回肠部位末检测到敏感乳杆菌(Lactobacillus agilis),回肠部位也末检测到约氏乳杆菌(Lactobacillus johnsonii)、不可培养细菌(Uncultured bacterium)等芽孢杆菌纲,而热应激不同时间段空肠和回肠部位可检测到不可培养细菌(Uncultured Escherichia sp)、溃疡拟杆菌(Bacteroides helcogenes)、卵形拟杆菌(Bacteroides ovatus)和索氏志贺氏菌(Shigella sonnei)等拟杆菌纲和γ-变形菌纲;其中约氏乳杆菌、敏感乳杆菌数量变化在空肠部位减少最明显(P0.05),空肠和回肠卵形拟杆菌、不可培养的拟杆菌数量明显上升(P0.05)。热应激环境下蛋鸡空肠、回肠部位菌群多样性较为丰富,其抑制乳杆菌属、不可培养细菌的增殖,促进卵形拟杆菌的繁殖,而促进卵形拟杆菌的繁殖,导致消化道菌群平衡的破坏。     

Influence of nitrogen sources on the tolerance of Lacticaseibacillus rhamnosus to heat stress and oxidative stress

It has been found that 32 genes related to nitrogen source metabolism in Lacticaseibacillus rhamnosus are downregulated under both heat stress and oxidative stress. In this study, the influence of different nitrogen sources within the growth medium on the tolerance of L. rhamnosus to heat stress and oxidative stress was investigated. Tryptone-free MRS was found to enhance the tolerance of L. rhamnosus hsryfm 1301 to heat stress and oxidative stress during the whole growth period, and this result was universal for all L. rhamnosus species analyzed. The strongest strengthening effect occurred when the OD₆₀₀ value reached 2.0, at which the survival rates under heat stress and oxidative stress increased 130-fold and 40-fold, respectively. After supplementing phenylalanine, isoleucine, glutamate, valine, histidine, or tryptophan into the tryptone-free MRS, the tolerance of L. rhamnosus to heat stress and oxidative stress exhibited a sharp drop. The spray drying survival rate of L. rhamnosus hsryfm 1301 cultured in the tryptone-free MRS rose to 75% (from 30%), and the spray dried powder also performed better in the experimentally simulated gastrointestinal digestion. These results showed that decreasing the intake of amino acids is an important mechanism for L. rhamnosus to tolerate heat stress and oxidative stress. When L. rhamnosus is cultured for spray drying, the concentration of the nitrogen source''s components should be an important consideration.     

Lethal and sublethal behavioural responses of saline water beetles to acute heat and osmotic stress

1. As species' physiological breadth determines their potential to deal with environmental changes, and influences individuals' survival and the persistence of populations, information about lethal and sublethal responses could be fundamental for conservation purposes. 2. We used a standard experimental approach to explore mortality and behavioural avoidance responses (i.e. flight and emersion from the water) to a combination of acute heat and osmotic stress on six species of saline water beetles (belonging to Enochrus, Nebrioporus, and Ochthebius genera). 3. Heat stress affected survival and behavioural responses in all of the species, whereas osmotic stress and the interaction between both stressors only showed significant effects for the Ochthebius genus. Behavioural and survival patterns were highly interrelated across the stress gradients. The Enochrus and Nebrioporus studied species showed maximum avoidance activity at 35–40 °C, and a short (< 30 min) exposure to 45 °C was lethal. Ochthebius species were the most heat tolerant and displayed increasing behavioural responses with increasing temperature. In the Nebrioporus and Ochthebius genera, the species occupying lotic, more environmentally stable habitats, showed greater mortality, and avoidance responses were higher or initiated at lower stress thresholds than lentic species. In contrast, both Enochrus species displayed a similar mortality, and the lentic species E. bicolor emerged and flew more than the lotic E. falcarius, in concordance with its higher dispersal capacity. 4. Avoidance responses could provide interesting information about species' physiological amplitudes as a complement to lethal responses. The lotic species here studied showed narrower physiological amplitude (i.e. N. baeticus and O. glaber) or lower dispersal ability (i.e. E. falcarius) than their lentic relatives; both traits could result in a higher vulnerability of lotic species to thermal habitat changes.