Effect of long-term heat exposure on peripheral concentrations of heat shock protein 70 (Hsp70) and hormones in laying hens with different genotypes

1. The effect of long-term, moderate heat stress (30 to 32 degrees C) on heat shock protein 70 (Hsp70) concentration in mononuclear blood cells and plasma concentrations of 3,5,3'-triiodothyronine (T3) and corticosterone in laying hens was investigated. 2. Three groups of 48 hens each (Ethopian line [Angete Melata, Na], New Hampshire [NH], F1 cross [Na x NH]) were divided into an experimental group (24 each) and a control group (24 each, ambient temperature 18 to 20 degrees C), respectively. All hens were kept in individual cages up to an age of 68 weeks and performance data were recorded. 3. Blood samples were taken from the wing vein of 12 hens from each group at weeks 22, 38, 51 and 65 (12 hens x 3 lines x 2 treatments). Mononuclear blood cells were isolated and Hsp70 concentrations were determined by Western Blot analysis with a monoclonal anti-Hsp70 antibody. T3 and corticosterone were measured with commercially available ELISA and RIA kits, respectively. 4. The moderate heat stress caused significantly increased Hsp70 levels compared with the control groups in weeks 51 and 65. However, the responses of the lines were not uniform at different ages. 5. In contrast, T3 levels were significantly decreased in stressed birds regardless of line and age. There was no effect of treatment and line on corticosterone levels during the experimental period. 6. Our results indicate that Hsp70 and T3 levels are affected by mild heat stress applied over a long period but are both involved in independent mechanisms of acquisition of thermotolerance. Further investigations are necessary to clarify whether the observed differences in Hsp70 response between the genotypes are indicators for differences in thermotolerance.     

Effects of heat stress on growth, some blood variables and lipid oxidation in broilers exposed to high temperature at an early age

1. This study was conducted to determine metabolic and physiological responses of 2 commercial broiler strains, Hubbard (H) and Cobb (C), exposed to an ambient temperature of 38 degrees +/- 1 degree C for 2 h at 14 and 15 d of age. 2. Exposure to high temperature at an early age resulted in weight loss in strain C, which was not compensated for by 35 d of age but there was no weight loss in strain H. 3. Exposure of broilers to heat stress (38 degrees +/- 1 degree C) at 35 d of age resulted in an increase in rectal temperature, regardless of previously high temperature experience but acid-base balance and haematocrit values were not affected by heat stress. 4. Malondialdehyde concentration was higher in unexposed birds than in previously exposed ones and did not significantly differ between strains.     

The effect of early-age food restriction on heat shock protein 70 response in heat-stressed female broiler chickens

1. This study was conducted to determine the effect of early-age food restriction on heat shock protein (hsp) 70 synthesis in the brains of female broiler chickens exposed to high ambient temperatures. 2. Chicks were brooded for 3 weeks and then maintained at 24+/-1 degrees C. 3. On d 0, chicks were assigned to one of 4 feeding regimens; each regimen was applied to 4 cages of chicks. The regimens were: (1) ad libitum feeding (AL); (2) 80% food restriction at 4, 5 and 6 d of age (F80); (3) 60% food restriction at 4, 5, and 6 d of age (F60); and (4) 40% food restriction at 4, 5 and 6 d of age (F40). From d 35 to d 41, all chicks were subjected to 38+/-1 degrees C for 2 h/d. 4. One day following food restriction (d 7), hsp 70 expression in the brain samples of F60 and F40 chicks was augmented but not those fed AL and F80. 5. Prior to the heat challenge (d 35), all chicks had similar hsp 70 response. Irrespective of feeding regimen, there was a marked increase in hsp 70 expression after 4 d of heat treatment (d 38). Following 7 d of heat exposure (d 41), except for the F60 chicks, the augmented hsp 70 expression in the brains of AL, F80 and F40 birds was not maintained. 6. Enhancement of hsp 70 expression was noted in birds subjected to F60, but not AL, F80 or F40, throughout the period of heat exposure.     

Effect of acute heat stress on heat shock protein 70 messenger RNA and on heat shock protein expression in the liver of broilers

1. The synthesis of heat shock protein 70 (Hsp70) mRNA and the expression of Hsp70 in the liver of broiler chickens submitted to acute heat stress (35°C for 5 h) was investigated.

2. Hsp70 expression was detected by SDS‐PAGE and Western blot analysis using a polyclonal antiserum against Hsp70 of Blastocladiella emersonii. The specific signal of Hsp70 mRNA was analysed by Northern blot using as probe a Hsp70 cDNA of B. emersonii.

3. An increase in the amount of Hsp70 was detected from the first up to the fifth hour of acute heat exposure. This increase in the amount of Hsp70 was accompanied by an increase in Hsp70 mRNA which peaked at 3 h.

4. This study shows that the heat induced increase in Hsp70 mRNA and protein in broiler liver, in vivo, are time dependent, similar to that in mammals.     

Effects of temperature treatments on the heat production of starving chickens

1. Heat production was measured for about 24 h at six temperatures from 2 to 35 degrees C on individually starved broilers that had been subjected to four treatments; these were acclimated or unacclimated to these temperatures, or to these temperatures for 12 h and to 22 degrees C for 12 h (alternated) during each 24-h period. 2. Response curves relating heart production and environmental temperature for the four different treatments differed significantly. Only the unacclimated birds subjected to the alternated temperatures increased heat production at 35 degrees/22 degrees C. Major effects of acclimation were observed mainly in the cold. 3. The relationship between daily endogenous nitrogen (N) excretion and heart production (mg N/kJ) was constant at the different temperatures, but acclimation and alternating temperature increased N excretion. 4. Evaporative heat loss was reduced by alternating temperature at the high temperatures, and by maintaining temperature constant in the cold.     

Alleviation of acute heat stress by food withdrawal or darkness

1. Growing 33-d-old broiler fowl were exposed to air temperatures of 35 to 40 degrees C for a period of 4 h each day for 5 d. 2. The birds were fed on diets providing widely differing proportions and intakes of energy as protein, carbohydrate and fat. 3. Three lighting and feeding treatments were superimposed on the 4-h heat stress period. The treatments were: LF (control) lights and food throughout, NL (no light) extinguished throughout the hot period, NF (no food) food withdrawn 2 h before the hot period and replaced immediately after it. 4. Diet had no significant effect on rate of increase of rectal temperature (TR) during heat stress. 5. The NL and NF treatments significantly reduced the rate of increase of TR and the TR attained after 4 h of heat exposure.     

热应激处理鼠囊胚Hsp70基因表达的RT-PCR检测

将体外培养的小鼠囊胚分别施以40℃1、2、3 h和38℃1、2、3 h热应激处理,然后在37℃条件下分别恢复3、2、1 h后,提取胚胎细胞总RNA,用RT-PCR方法检测小鼠囊胚Hsp70基因的表达,以明确小鼠囊胚经热应激处理后是否有诱导型Hsp70基因表达。结果表明:各试验组,包括对照组鼠的囊胚都有诱导型Hsp70基因的表达。说明小鼠囊胚对热应激处理敏感,轻微热应激就可使诱导型Hsp70基因表达。     

Effect of Heat Stress on Milk Production,Rectal Temperature,Respiratory Rate and Blood Chemistry in Holstein,Jersey and Australian Milking Zebu Cows

The effect of heat stress on changes in milk production, rectal temperature, respiratory rate and blood chemistry was evaluated in three groups of six mature Holstein, Jersey and Australian Milking Zebu (AMZ) dairy cows. These animals were subjected to a cool environment when the mean temperature-humidity index (THI) was 72+/-1.4 (dry bulb temperature of 22.2-24.4 degrees C and relative humidity of 100-60%) during the month of December. This experiment was repeated during the hotter month of July of the following year, when the mean THI was 93+/-3.1 (dry bulb temperature of 35.6-43.9 degrees C and relative humidity 95-35%). Holstein cows produced more (p <0.01) milk than AMZ and Jersey cows during the cooler months of the year and all the cows were dry during the hotter months from June until September. Heat stress increased (p<0.01) rectal temperature and respiratory rate in all three breeds. Heat stress had no effect on blood pH in Holstein and AMZ cows but lowered (p <0.01) blood pH from 7.42 to 7.34 in Jersey cows. In addition, heat stress lowered (p <0.01) blood pCO2 (kPa), bicarbonate (HCO3, mmol/L), base excess (BE, mmol/L) and plasma chloride (Cl-, mmol/L) in all three breeds. The total haemoglobin (THb, g/dl) was elevated (p <0.01) in all three breeds when they were subjected to heat stress. Heat stress increased (p<0.01) oxygen saturation (O2SAT, %) in Jersey and AMZ cows but lowered it (p <0.01) in Holstein cows. On the other hand, heat stress increased (p <0.01)pO2 (kPa) in Holstein and Jersey cows but lowered it (p <0.01) in AMZ cows. Heat stress increased (p <0.01) plasma potassium (K, mmol/L) and calcium (Ca, mmol/L) only in Holstein and Jersey cows but lowered them (p<0.01) in AMZ cows. The plasma glucose (GLU, mmol/L) increased (p<0.01) with heat stress in Holstein and AMZ cows but decreased (p <0.01) in Jersey cows. Heat stress increased (p<0.01) plasma creatinine (CR, (mol/L) but lowered (p<0.01) plasma creatinine phosphokinase (CPK, IU/L), aspartate aminotransferase (AST, IU/L) and blood urea nitrogen (BUN, mmol/L) in all three breeds. These results indicate that heat-stressed Holstein and AMZ cows were able to maintain their acid-base balance with a marginal change in their pH of 0.02 when their rectal temperatures increased by 0.47 and 0.38 degrees C, respectively. When heat stress increased the rectal temperature in Jersey cows by 0.70 degrees C, the pH decreased (p<0.01) from 7.42 to 7.34. However, even with this decrease 0.08 the pH is still within the lower physiological limit of 7.31.     

Responses to heat stress in commercial and local broiler stocks. 1. Performance traits.

1. This study investigated effects of conditioning at 5 d of age and food restriction during a hot period on responses to prolonged heat stress of two fast (G1 and G2) and one slower-growing (G3) broiler stocks. 2. Chicks from each stock were divided into 3 groups: control, conditioned (chicks exposed to 36 degrees C for 24 h at 5 d of age) and food restricted (during the heat stress food was withdrawn 2 h before the hot period and chicks were fed between 17.00 and 08.00 h). Prolonged heat stress was induced daily from 28 to 49 d by heating until the ambient temperature reached 32 degrees to 35 degrees C between 10.00 and 17.00 h. 3. Body weights, mortality and rectal temperatures were lower and food conversion higher for stock G3 than G1 or G2. 4. Management procedures used in this study improved food conversion and lowered mortality without affecting body weight. Rectal temperatures of the treated groups were similar to those of the controls.     

Absorption of protein, fatty acids and minerals in young turkeys under heat and cold stress

1. Absorption of protein, fatty acids, calcium, phosphate and potassium by young turkeys maintained at thermoneutral (24 degrees C), hot (35 degrees C) and cold (8 degrees C) conditions was examined. 2. Non-acclimatised, heat-stressed birds absorbed less potassium and phosphate, whereas absorption of nitrogen, fatty acids and calcium was not altered, as compared with birds at 24 degrees C. Non-acclimatised, cold-stressed birds absorbed less calcium than control birds and more nitrogen than non-acclimatised, heat-stressed birds. 3. Heat acclimatization might reduce the adverse effect of heat stress on potassium and phosphate absorption.     

Dietary supplementation of Zingiber officinale and Zingiber zerumbet to heat‐stressed broiler chickens and its effect on heat shock protein 70 expression,blood parameters and body temperature

The present study was conducted to assess the effects of dietary supplementation of Zingiber officinale and Zingiber zerumbet and to heat‐stressed broiler chickens on heat shock protein (HSP) 70 density, plasma corticosterone concentration (CORT), heterophil to lymphocyte ratio (HLR) and body temperature. Beginning from day 28, chicks were divided into five dietary groups: (i) basal diet (control), (ii) basal diet +1%Z. zerumbet powder (ZZ1%), (iii) basal diet +2%Z. zerumbet powder (ZZ2%), (iv) basal diet +1%Z. officinale powder (ZO1%) and (v) basal diet +2%Z. officinale powder (ZO2%). From day 35–42, heat stress was induced by exposing birds to 38 ± 1 °C and 80% RH for 2 h/day. Irrespective of diet, heat challenge elevated HSP70 expression, CORT and HLR on day 42. On day 42, following heat challenge, the ZZ1% birds showed lower body temperatures than those of control, ZO1% and ZO2%. Neither CORT nor HLR was significantly affected by diet. The ZO2% and ZZ2% diets enhanced HSP70 expression when compared to the control groups. We concluded that dietary supplementation of Z. officinale and Z. zerumbet powder may induce HSP70 reaction in broiler chickens exposed to heat stress.     

Respiratory alkalosis and primary hypocapnia in Labrador Retrievers participating in field trials in high-ambient-temperature conditions

OBJECTIVE: To determine whether Labrador Retrievers participating in field trials develop respiratory alkalosis and hypocapnia primarily in conditions of high ambient temperatures. ANIMALS: 16 Labrador Retrievers. PROCEDURES: At each of 5 field trials, 5 to 10 dogs were monitored during a test (retrieval of birds over a variable distance on land [1,076 to 2,200 m]; 36 assessments); ambient temperatures ranged from 2.2 degrees to 29.4 degrees C. For each dog, rectal temperature was measured and a venous blood sample was collected in a heparinized syringe within 5 minutes of test completion. Blood samples were analyzed on site for Hct; pH; sodium, potassium, ionized calcium, glucose, lactate, bicarbonate, and total CO2 concentrations; and values of PvO2 and PvCO2. Scatterplots of each variable versus ambient temperature were reviewed. Regression analysis was used to evaluate the effect of ambient temperature (< or = 21 degrees C and > 21 degrees C) on each variable. RESULTS: Compared with findings at ambient temperatures < or = 21 degrees C, venous blood pH was increased (mean, 7.521 vs 7.349) and PvCO2 was decreased (mean, 17.8 vs 29.3 mm Hg) at temperatures > 21 degrees C; rectal temperature did not differ. Two dogs developed signs of heat stress in 1 test at an ambient temperature of 29 degrees C; their rectal temperatures were higher and PvCO2 values were lower than findings in other dogs. CONCLUSIONS AND CLINICAL RELEVANCE: When running distances frequently encountered at field trials, healthy Labrador Retrievers developed hyperthermia regardless of ambient temperature. Dogs developed respiratory alkalosis and hypocapnia at ambient temperatures > 21 degrees C.     

Evaluation of four warming procedures to minimise heat loss induced by anaesthesia and surgery in dogs

OBJECTIVE: To evaluate the efficiency of four warming procedures, introduced after anaesthetic induction and continued during surgery, in minimising heat loss in anaesthetised dogs. DESIGN: Dogs were paired. One of each pair was a control; the other was subjected to one of four warming procedures. METHODS: Ninety-six dogs were involved in total. Pairs of dogs were matched for breed, hair length, and type of surgical procedure and placed adjacent to each other in a large temperature-controlled surgical theatre. One dog within each pair was assigned to one of four warming procedures that commenced immediately after anaesthetic induction. Group 1 (11 pairs) were placed on a purpose-designed prewarmed (41 degrees C) electrically heated pad. Group 2 (18 pairs) were placed on a prewarmed electric heat pad (41 degrees C), cocooned by four wrapped water bottles (initially 41 degrees C) and subjected to radiant heat (150 watt lamp placed 50 cm away from the head of the dog). Group 3 (11 pairs) were surrounded by a forced air warming mattress (set at 43 degrees C). Group 4 (8 pairs) were connected via the anaesthetic breathing circuit to a heater/humidifier (set at 41 degrees C). Rectal temperature measurements were recorded every 15 min for the first 3 h of anaesthesia. The fall in rectal temperature of the control dog was subtracted from the fall in temperature of the treatment dog and this measurement was used to assess the efficacy of the various warming procedures. RESULTS: The mean rectal temperature of unheated 'control' dogs decreased 1.9 +/- 0.6, 1.4 +/- 0.4 and 1.1 +/- 0.4 degrees C over the first, second and third hour respectively. After 3 h the temperature fall differential for all groups were 0.7 +/- 0.7 (Group 1), 3.1 +/- 1.1 (Group 2), 2.4 +/- 1.1 (Group 3) and 1.0 +/- 1.1 degrees C (Group 4). Thus the group 2 procedure was the most successful in preventing a drop of temperature followed by groups 3, 4 and 1. CONCLUSION: Large dogs undergo significant reduction in core body temperature especially during the first 2 h of anaesthesia and surgery. Supplementary warming substantially reduces this fall in body temperature, although certain warming procedures were found to be more effective than others.     

Hepatic tyrosine aminotransferase activity is affected by chronic heat stress and dietary tyrosine in broiler chickens

1. Two experiments were conducted to investigate the impact of high temperature and dietary tyrosine (Tyr) content on performance and activity of hepatic tyrosine aminotransferase (EC 2.6.1.5.), an enzyme that catalyses the first step in the metabolic degradation of Tyr in broiler chickens. 2. Two-week-old birds were allocated to one of three temperature treatments: 24 degrees C (control), 36 degrees C (heat stress, HS) and 24 degrees C pair-fed (24PF) for 2 weeks and fed on diets containing 100% (Experiment 1) and 50, 100 and 200% (Experiment 2) of the NRC requirement for Tyr. 3. In Experiment 1, exposure of chickens to 36 degrees C for 2 weeks caused significant increase in hepatic tyrosine aminotransferase activity but no significant change in activity of hepatic phenylalanine 4-hydroxylase (EC 1.14.16.1) (an enzyme that catalyses conversion of phenylalanine to Tyr) compared with the 24PF birds. No significant changes attributable to heat stress were detected in hepatic glutamic-oxaloacetic transaminase (EC 2.6.1.1) activity. 4. In Experiment 2, heat stress caused reductions in weight gain and feed intake in chickens on all diets, compared with their control counterparts. Hepatic tyrosine aminotransferase activity was increased by heat stress compared with their 24PF counterparts in chickens fed on the 100 and 200% Tyr diets, while in chickens fed the 50% Tyr diet, it was reduced by heat stress. 5. From these results, it is suggested that hepatic tyrosine aminotransferase activity is affected by heat stress and dietary Tyr content and the increased tyrosine aminotransferase activity with, in part, relatively low phenylalanine hydroxylase activity in hepatic tissues may be involved in the Tyr metabolism characteristic of heat-stressed chickens.     

Behavioural and physiological responses of three chicken breeds to pre-slaughter shackling and acute heat stress

1. The aim of this study was to compare the behavioural and physiological responses to hanging and acute heat stress in three different chicken breeds. Chicks were obtained from a slow-growing French 'Label Rouge' line (SGL), a fast-growing standard line (FGL) and a heavy line (HL). The SGL, FGL and HL birds were slaughtered at their respective market ages of 12, 6 and 6 weeks, in an attempt to achieve similar body weights. Before stunning, birds were either shackled by their legs on the moving line for 2 min (shackling stress: SH) or placed in a room at 35 degrees C and 60% of humidity for 3.5 h and then shackled for 2 min (acute heat stress plus shackling: H + SH) or subjected to minimal stress by shackling for 10 s before stunning (control group: C). 2. Bird physiological responses to the three pre-slaughter treatments were estimated by measuring blood corticosterone, glycaemia, creatine kinase activity, acid-base status and electrolyte concentration as well as lactate content and glycolytic potential in the breast (Pectoralis major) and thigh (Ilio tibialis) muscles. Behavioural responses to shackling stress were evaluated by measuring wing flapping duration, straightening up attempts and vocalisations. 3. Blood corticosterone was higher in SH and H+SH groups than in the C group, regardless of genotype. The struggling activity on the shackle line differed among chicken breeds. It was more intense and occurred more rapidly after hanging in the SGL birds than in both other breeds. Furthermore, SGL struggling activity was not affected by hanging duration while it increased with hanging duration in FGL and HL birds. 4. Wing flapping duration was negatively correlated with blood pH, bicarbonate concentration and positively correlated with breast muscle lactate content, indicating that struggling stimulated antemortem glycolysis activity in breast muscle. Acute heat stress affected blood Ca2+ and Na+ concentration and increased glycaemia and glycolytic potential of thigh muscle. 5. Both acute heat stress and shackling before slaughter were experienced as stressful events by all types of birds.     

Comparative studies on temperature threshold for heat shock protein 70 induction in young and adult Murrah buffaloes

To know the temperature threshold for heat shock protein 70 (HSP70) induction in lymphocytes and to assess physiological changes, if any, in relation to HSP70 induction in young and adult Murrah buffaloes, this study was divided into two parts: I. In vivo study: where assay of HSP70 was performed in blood samples collected from acutely exposed young and adult Murrah buffaloes (n = 6) inside a climatic chamber at 40, 42 and 45 °C for 4 h and thermoneutral temperature (22 °C). Physiological parameters viz., rectal temperature, respiratory rate, pulse rate and skin temperature of different body parts were monitored to assess magnitude of stress in the animals owing to thermal exposure II. For in vitro study, equal numbers of lymphocyte cells were separated from blood collected from young and adult buffaloes and were subjected to four temperature treatments (38, 40, 42 and 45 °C) for 4 h. A significant increase (p < 0.05) in all the physiological parameters in both young and adult buffaloes was observed after exposure to 40, 42 and 45 °C for 4 h as compared to 38 °C. The average plasma HSP70 concentrations (ng/ml) were significantly higher (p < 0.05) at 40, 42 and 45 °C as compared to 38 °C in both young and adult and were higher in young than adult buffaloes at 38 and 45 °C. Heat shock protein 70 level in lymphocyte lysate showed highest concentration after 3-h exposure to all temperatures (40, 42 and 45 °C) in both young and adult buffaloes. The intensity of changes of all physiological parameters was more in young animals than in the adults indicating the greater susceptibility of younger animals to heat stress and was found to be changed at around 40 °C when animals were exposed to different temperatures, indicating the possibility that HSP70 production may be initiated at this temperature which is 2 or 3 °C higher than core body temperature.     

Epigallocatechin-3-gallate supplementation can improve antioxidant status in stressed quail

1. Heat stress causes oxidative stress, which decreases plasma antioxidants in poultry. Epigallocatechin gallate (EGCG), the most abundant polyphenol in green tea, is a powerful antioxidant against lipid peroxidation. This experiment was conducted to evaluate the effects of dietary EGCG supplementation on performance, carcase characteristics, concentrations of malondialdehyde, lipid peroxidation indicator, vitamins C, E, A, cholesterol, triglyceride, and glucose in Japanese quail (Coturnix coturnix japonica) exposed to a high ambient temperature of 34 degrees C. 2. One hundred and eighty Japanese quails (10 d old) were assigned at random to 6 treatment groups consisting of 10 replicates of three birds. Birds were kept in cages in a temperature-controlled room at either 22 degrees C (thermo-neutral) or 34 degrees C (heat stress) for 8 h/d. Birds were fed either a basal diet or the diet supplemented with 200 or 400 mg of EGCG/kg of diet. 3. EGCG supplementation linearly increased feed intake, live weight gain, feed efficiency, cold carcase weight and yield under heat stress conditions but did not show the same effect at thermoneutral conditions. Serum vitamin C, E, and A concentrations increased in birds reared at high temperature while non-significant changes occurred in thermo-neutral groups. Malondialdehyde concentrations in serum and liver decreased in all birds of both thermo-neutral and heat stressed groups as dietary EGCG supplementation increased. Heat stress-induced increase in serum cholesterol, triglycerides and glucose concentrations were linearly reversed by EGCG supplementation. 4. The results indicate that EGCG supplementation improved the live performance and antioxidant status of heat-stressed Japanese quail.     

Identification of the heat shock protein 70 (HLHsp70) in Haemaphysalis longicornis

A Haemaphysalis longicornis heat shock protein 70 (HLHsp70) was identified from a cDNA library synthesized from tick eggs. The HLHsp70 cDNA is 2311 bp in length and encodes 661 amino acid residues with the predicted molecular weight of 72.5 kDa and an isoelectronic point (pI) of 5.2. It also contains the highly conserved functional motifs of the Hsp70 family and a specific endoplasmic reticulum (ER) retention signal "KDEL" that is common among ER-localized proteins. The HLHsp70 exhibits 90% amino acid identity to the putative Hsp70 of Ixodes scapularis, and 85% to Gallus gallus 78 kDa glucose-regulated protein precursor. Real time RT-PCR analysis showed that the expression levels of the Hsp70 in ovaries and salivary glands were significantly higher than in other tested tissues in partially fed females. Although the expression level of the HLHsp70 was constantly low in unfed ticks, it was significantly induced by blood-feeding. Further, the expression was positively correlated to the temperature (4-37°C, tested). Western blot analysis showed that the rabbit antiserum against the recombinant HLHsp70 protein (rHLHSP70) recognized bands of approximately 100, 72, and 28 kDa from egg lysates, as well as a 72kDa fragment in protein extracts from partially fed larvae. Immunization of rabbits with the rHLHSP70 did not result in a statistically significant reduction of female tick engorgement and oviposition. These results suggest that although HLHSP70 plays a role in the physiological activities of ticks, as a constitutive protein it was not suitable for selection as a candidate vaccine antigen against ticks.     

家蚕热休克蛋白70家族基因的染色体定位及表达特征

热休克蛋白70家族(Hsp70)是非常保守的蛋白家族,每个物种的Hsp70家族都有多个成员,每个成员都可能具有特殊的功能。为了能够对家蚕Hsp70家族各成员进行科学命名和了解其特有的功能,分析家蚕Hsp70家族成员的编码基因及其在染色体上的定位,并通过实时荧光定量PCR方法检测这些基因在5龄第3天幼虫中肠、脂肪体和丝腺组织中的转录表达情况。家蚕诱导型Hsp70家族成员Bmhsp70A和Bmhsp70B的编码基因集中分布在第27号染色体上,有多个拷贝;组成型Bmhsp70家族成员的编码基因则分布在不同的染色体上,一般仅有1个拷贝。家蚕Hsp70家族基因中有10个能转录并翻译蛋白质产物的成员,还有1个能正常转录但不能正确翻译的假基因。实时荧光定量PCR结果表明:在常温(25℃)条件下,组成型Bmhsp70家族基因Bmhsc70-4和Bmhsc70-3的表达水平较高,而其它基因成员的表达水平则相对较低,诱导型Bmhsp70家族基因成员在常温下也有一定程度的基础表达;40℃热激2 h后,Bmhsp70家族所有基因的表达水平都有所升高,其中诱导型Bmhsp70家族基因Bmhsp70B和Bmhsp68的转录表达上升水平远高于组成型Bmhsp70家族基因Bmhsc70-3和Bmhsc70-4。不同表达类型的家蚕Hsp70家族基因成员在5龄幼虫中肠、丝腺和脂肪体中的表达情况不同,热激2 h后的表达变化规律也不完全相同,提示家蚕Hsp70家族成员具有各自特殊的功能。