温度应激下缢蛏血淋巴蛋白质组的比较研究

缢蛏(Sinonovaculaconstricta,Lamarck)历来是我国沿海重要的经济养殖贝类。本文应用蛋白质组学方法探求其在高温环境应激下血淋巴内40000U以下小分子蛋白质组的变化,寻找缢蛏血淋巴的抗性机制。在高温应激下,缢蛏血淋巴内有多种蛋白发生变化,最终鉴定出2种蛋白质:锌指蛋白及IBP-CARCRchelonianin。本研究也初步证明了蛋白质组理论及技术在分析生态学问题时的适用性及灵活性。     

Effect of thermal stress,restricted feeding and combined stresses (thermal stress and restricted feeding) on growth and plasma reproductive hormone levels of Malpura ewes under semi‐arid tropical environment

A study was conducted to assess the effect of thermal, nutritional and combined stresses (thermal and nutritional) on the growth, oestradiol and progesterone levels during oestrus cycles in Malpura ewes. Twenty‐eight adult Malpura ewes were used in the present study. The ewes were randomly allocated into four groups, viz., GI (n = 7; control), GII (n = 7; thermal stress), GIII (n = 7; restricted feeding) and GIV (n = 7; combined stress). The animals were stall fed with a diet consisting of 60% roughage and 40% concentrate. GI and GII ewes were provided with ad libitum feeding while GIII and GIV ewes were provided with restricted feed (30% intake of GI and GII ewes) to induce nutritional insufficiency. GII and GIV ewes were kept in climatic chamber at 40°C and 55% RH for 6 h a day between 10:00 and 16:00 hours to induce thermal stress for a period of two oestrous cycles. Parameters studied were body weight, oestrus incidences, plasma oestradiol 17‐β, plasma progesterone, conception rate, gestation period, lambing rate, and birth weight of lambs. The results indicate that combined stress significantly (p < 0.05) reduced body weight, oestrus duration, birth weight of lambs, and oestradiol 17‐β whereas significantly (p < 0.05) increased oestrus cycle length and progesterone. Furthermore, the results reveal that on comparative basis, ewes were able to better adapt in terms of growth and reproduction to restricted feeding than thermal stress. However, when restricted feeding was coupled with thermal stress it had significant (p < 0.05) influence on body weight, average daily gain, oestradiol 17‐β and progesterone concentrations. This showed that combined stress were more detrimental for these reproductive hormones in Malpura ewes under a hot semi‐arid environment.     

Optimal dietary lysine to threonine ratio in pigs (30–110 kg BW) derived from observed dietary amino acid efficiency

Optimal ratio of lysine (Lys) to threonine (Thr) in diets for growing barrows [genotype: Piétrain × (Large White × German Landrace)] was established at 30, 50, 70, 90 and 110 kg body weight (BW). N balance studies were conducted with diets based on constant mixture of wheat, barley, soybean meal and field peas. Supplementation of crystalline amino acids (AA) provided diets with Lys (diet A) and Thr (diet B) in first limiting position. For each BW range, totally 28 individual N balance data were utilized for assessing model parameters (exponential N utilization model) of dietary efficiency of Lys and Thr, respectively. Observed maximal dietary efficiency of Lys and Thr provided the database for conclusion of optimal dietary Lys to Thr ratio. Based on applied experimental conditions, BW dependent Lys to Thr ratio could not be concluded. On average, the dietary ratio Lys to Thr = 1:0.61 ± 0.02 was obtained as optimal for growing pigs.     

Effect of arginine supplementation on the morphology and function of intestinal epithelia and serum concentrations of amino acids in pigs exposed to heat stress

The exposure of pigs to heat stress (HS) appears to damage their intestinal epithelia, affecting the absorption of amino acids (AAs). Arg is involved in the restoration of intestinal epithelial cells but HS reduces Arg intake. The effect of dietary supplementation with Arg on the morphology of intestinal epithelia, AA transporter gene expression, and serum concentration (SC) of free AAs in HS pigs was analyzed. Twenty pigs (25.3 ± 2.4 kg body weight) were randomly assigned to two dietary treatments: Control (0.81% Arg), wheat–soybean meal diet supplemented with l-Lys, l-Thr, dl-Met, and l-Trp, and the experimental diet where 0.16% free l-Arg was supplemented to a similar Control diet (+Arg). All pigs were individually housed and exposed to HS, fed ad libitum with full access to water. The ambient temperature (AT), recorded at 15-min intervals during the 21-d trial, ranged on average from 29.6 to 39.4 °C within the same day. Blood samples were collected on day 18 at 1600 hours (AT peak); serum was separated by centrifugation. At the end of the trial, five pigs per treatment were sacrificed to collect samples of mucosa scratched from each small intestine segment. The expression of AA transporters in intestinal mucosa and the SC of AAs were analyzed. Villi height (VH) was higher (P < 0.01) in the duodenum, jejunum, and ileum but the crypt depth did not differ between the Control and the +Arg pigs. Supplementation of l-Arg increased the mRNA coding for the synthesis of the cationic AA transporter b^0,+ (P < 0.01) and the neutral AA transporter B⁰ (P < 0.05) in the duodenum by approximately 5-fold and 3-fold, respectively, but no effect on mRNA abundance was observed in the jejunum and ileum. The supplementation of l-Arg increased serum Arg, His, Met, Thr, Trp, and urea (P < 0.05) and also tended to increase Val (P < 0.10) but did not affect Ile, Lys, Leu, and Phe. These results indicate that supplementing 0.16% l-Arg to the Control diet may help to improve the function of the small intestine epithelium, by increasing the VH, the abundance of AA transporters, and the SC of most indispensable AAs in pigs exposed to HS conditions. However, the lack of effect of supplemental Arg on both Lys SC and weight gain of pigs suggests that increasing the Lys content in the +Arg diet might be needed to improve the performance of HS pigs.     

The effects of early‐age thermal manipulation and daily short‐term fasting on performance and body temperatures in broiler exposed to heat stress

This study was conducted to investigate the effects of thermal manipulation at 5 days of age and short‐term fasting during the warmest part of the day on responses to prolonged heat stress of broilers. A total of 240‐day‐old Ross 308 female broiler chicks were divided into three groups: control, thermal manipulation (chicks were exposed to 36 °C for 24 h at 5 days of age) and short‐term fasting during the warmest part of the day (10.00–17.00 h). Prolonged heat stress was induced daily from 28 to 42 days by heating until the ambient temperature reached 32–35 °C between 10.00 and 17.00 h. Both thermal manipulation and short‐term fasting resulted in a decrease in rectal temperatures and haematocrit values at 35 and 41 days of age. Thermal manipulation improved body weight, feed consumption and feed conversion. However, short‐term fasting caused a reduction in body weight and a deterioration in feed conversion. Short‐term fasting lowered the percentages of carcass, whereas thermal manipulation highered breast yield. Both thermal manipulation and short‐term fasting decreased heart mass and abdominal fat.     

高温胁迫下粤选1号匍匐翦股颖新品系的生理效应研究

对匍匐翦股颖(Agrostis stolonifera)品种Penncross及其所选育出的粤选1号新品系在高温胁迫条件下的一些生理效应进行研究,结果表明,在高温胁迫下,匍匐翦股颖原品种叶片中的丙二醛(MDA)含量随高温胁迫时间延长而持续增加,新品系MDA的含量在胁迫初期增加,随后呈下降至缓慢上升趋势.新品系叶片中游离脯氨酸的含量在高温胁迫下显著增加,而原品种的在高温胁迫第3天才开始显著增加.高温胁迫下,新品系游离脯氨酸含量明显高于原品种.新品系SOD,POD和CAT活性在胁迫第1天便显著增加,而原品种的保护酶活性到胁迫第4天才显著增加.此外,新品系叶片中SOD,POD和CAT活性远高于原品种,这可能与新品系具有较强的应急机制有关.     

温度胁迫对亚热带常用草坪草活性氧代谢相关酶的影响

研究了在高温和低温胁迫下,亚热带建坪常用草坪草叶片的活性氧代谢相关酶与细胞膜透性的变化,并依据"自由基"假说,探讨了温度胁迫条件下自由基对草坪草细胞(主要是细胞膜体系)的伤害机理,初步揭示了不同温度胁迫下草坪草受害的致成原因以及不同品种间的差异.研究结果表明,以活性氧代谢指标与细胞膜透性为评判抗性大小的依据,各类品种的抗寒性大小顺序是匍匐剪股颖Agrostis stolonifere>草地早熟禾Poa pratensis>多年生黑麦草Lolium perenne>高羊茅Festuca arundinacea>狗牙根Cynodon dactylon,抗热性大小顺序是狗牙根 > 高羊茅 > 草地早熟禾 > 多年生黑麦草 > 匍匐剪股颖.研究运用了亚热型草坪草活性氧代谢相关酶测定的方法进行了草坪草品种鉴定和抗性比较,为亚热带地区草坪草品种选择、建植和维护提供了科学的理论依据.     

碱胁迫下星星草的主要胁变反应

用0～200mmol／L的碱性盐Na2CO3对星星草进行胁迫处理，观测了星星草对碱胁迫的主要生理反应。实验结果表明：随着Na2CO3胁迫浓度的增大，星星草体内总有机酸、柠檬酸和脯氨酸含量明显上升，含水量、总氮含量和叶绿素含量明显下降，Na^ 含量上升，K^ 含量下降，K^ ／Na^ 降低。胁迫浓度的增加还使植物体的根系活力下降，质膜透性增加。各胁变指标的最大变化多发生在150mmol／L Na2CO3浓度附近。     

Relationship of blood corticosterone, immunoglobulin and haematological values in young crocodiles (Crocodylus porosus) to water temperature, clutch of origin and body weight

Objective To examine whether sub-optimal temperature induced stress and immunosuppression in farmed saltwater crocodile (Crocodylus porosus) hatchlings. Design A clinico-pathological study. Animals A total of 140 hatchlings were used. Procedure Body weight and length, plasma corticosterone and immunoglobulin concentrations and total and differential white blood cell counts were measured in 140 hatchlings from five clutches divided between five water temperature treatment groups. Initially all groups were housed at 32°C for 10 weeks, then two groups (L, LC) were changed to low temperature (28°C) and two groups (H, HC) to high temperature (36°C), while one group (C) remained at 32°C. The LC and HC groups were maintained at these temperatures for 10 days, after which the water temperature of both groups was returned to 32°C. Blood samples were collected twice (at 6 and 9 weeks of age) before the initial temperature change, and at 10 days and 4 weeks after the initial temperature change (at 11.5 and 14 weeks of age). Results Except for an increase in plasma corticosterone in the HC group and a decrease in the L group when the temperature change was first introduced, changes in plasma corticosterone were not significant. There were no significant changes in immunoglobulin concentrations. There were, however, significant decreases in the total white cell and lymphocyte counts in the LC group after the temperature was decreased to 28°C, and an increase in these counts after water temperature was returned to 32°C. Clutch of origin had significant effects on body weight and length gains, and there were negative relationships between body weight and corticosterone concentrations and between body weight and immunoglobulin concentrations. Conclusions As haematological changes indicative of stress were not associated with significant changes in serum corticosterone, immunosuppression in young crocodiles may be independent of the hypothalamic-pituitary-adrenal cortical axis.     

Effect of epidermal growth factor (EGF) on the phosphorylation of mitogen-activated protein kinase (MAPK) in the bovine oviduct in vitro: Alteration by heat stress

Epidermal growth factor (EGF) has been shown to be involved in control of the oviductal microenvironment. To elucidate the potential mechanisms responsible for the detrimental effect of heat stress and to identify the relation with the endocrine status, the effects of EGF on the level of phosphorylated mitogen-activated-protein kinase (MAPK) and proliferation of bovine oviductal epithelial cells (OECs) exposed to different cyclic ovarian steroidal environments (luteal phase (LP), follicular phase (FP) and postovulatory phase (PO)) and temperatures (mild heat stress (40 C) and severe heat stress (43 C)) were investigated. Western blot was performed to evaluate phosphorylated MAPK, while proliferation was analyzed by MTT assay. Stimulation of OECs with EGF alone or with EGF in the PO and FP environments significantly increased the amount of phosphorylated MAPK, with MAPK 44 phosphorylation being highest during exposure to PO conditions. These effects were not observed in the LP. Heat treatment completely blocked effects of EGF on phosphorylated MAPK. Additionally, severe heat stress led to a significantly lower basal level of phosphorylated MAPK. PD98059 (MAPK inhibitor) completely abolished EGF-stimulated MAPK phosphorylation and OECs proliferation. Overall the results indicate that EGF has the potential to increase the amount of phosphorylated MAPK in OECs and therefore could be involved in regulation of the bovine oviductal microenvironment. However, these regulatory mechanisms may be compromised in the presence of heat stress (high ambient temperature), leading to low fertility rates and impaired embryo survival.     

The effects of exposure duration to optimal concentration of 2‐phenoxyethanol on primary and secondary stress responses in kutum (Rutilus frisii kutum)

The goal of this study was to assess the anaesthetic induction and recovery time in kutum (Rutilus frisii kutum) after exposure to various concentrations (0.1, 0.3, 0.5, 0.7 and 0.9 ml/l) of 2‐PE as an anaesthetic, as well as the effects of optimal concentration (0.7 ml/l) of 2‐PE in relation to different exposure time (3, 10, 15 min) on some haematological and serum biochemical indices in this species. Moreover, the effects of 0.7 ml/l on blood parameters were assessed 24 h after the longest exposure. Significant increase was determined in Hb, MCH and MCHC after 10‐min exposure to 2‐PE (p < 0.05). Moreover, Hct, Hb and RBC levels increased significantly after 15 min‐exposure to 2‐PE (p < 0.05). There were no prominent changes in WBC and MCV. The plasma concentrations of glucose, cholesterol and cortisol increased significantly after 10‐ and 15‐min exposure to 2‐PE (p < 0.05) compared with the control group and all other exposure times. The activity of ALT and AST were significantly increased after 10‐ and 15‐min exposure respectively (p < 0.05). In this study, it appears that anaesthetizing kutum with 2‐PE at 0.7 ml/l for 3 min had no effect on the stress markers.     

Determination of carcinoembryonic antigen and cancer antigen (CA 15‐3) in bitches with tumours on mammary gland: preliminary report

The aim of this work was to determine levels of carcinoembryonic antigen (CEA) and cancer antigen (CA 15‐3) in the blood serum of 45 bitches. A modified procedure was used to determine the CEA and CA 15‐3 markers with the human kits using the radioimmunoassay method. Samples collected from extirpated tumour of mammary glands were histologically processed and classified as per WHO guidelines. The average age of animals with tumour was 10.00 ± 2.2 years; for healthy bitches average age was 4.2 ± 3.2 years. Values of CEA and CA 15‐3 were considered positive, if they exceeded 0.23 ng mL^?1 and 7 IU mL^?1, respectively. Average levels of CEA in the tumour group were 0.25 ± 0.06 versus 0.20 ± 0.03 in healthy bitches (P = 0.0001). The average CA 15‐3 value in bitches with tumour was 8.58 ± 1.27 versus 5.14 ± 1.34 in healthy animals (P < 0.0001).