Effect of Supplemental Trace Minerals on Hsp-70 mRNA Expression in Commercial Broiler Chicken

The effects of supplementing the organic forms of selenium (Se), chromium (Cr), and zinc (Zn) on Hsp-70 mRNA expression and body weight in broiler chickens were evaluated. 200 chicks were equally distributed into stainless steel battery brooders at the rate of 5 birds per pen and reared under heat stress condition up to 42^nd day. The chicks were fed with three experimental diets supplemented with organic forms of Se (0.30?mg/kg), Cr (2?mg/kg), and Zn (40?mg/kg) during the starter and finisher phases and a control diet without any supplementation. On the 21st and 42nd day, 20 birds from each period were sacrificed and samples were collected for analysis. Organic Se, Cr, and Zn supplementation significantly (P?Hsp-70 mRNA levels. The Hsp-70 mRNA expression levels were significantly (P?Hsp-70 mRNA expression level was not affected by age of the birds. The study concluded that organic trace mineral (oTM) supplementation resulted in low Hsp-70 mRNA expression, indicating reduced heat stress in broilers.     

Evaluation of DNA methylation and mRNA expression of heat shock proteins in thermal manipulated chicken

Thermal manipulation during embryogenesis has been demonstrated to enhance the thermotolerance capacity of broilers through epigenetic modifications. Heat shock proteins (HSPs) are induced in response to stress for guarding cells against damage. The present study investigates the effect of thermal conditioning during embryogenesis and thermal challenge at 42 days of age on HSP gene and protein expression, DNA methylation and in vitro luciferase assay in brain tissue of Naked Neck (NN) and Punjab Broiler-2 (PB-2) chicken. On the 15th day of incubation, fertile eggs from two breeds, NN and PB-2, were randomly divided in to two groups: control (C)—eggs were incubated under standard incubation conditions, and thermal conditioning (TC)—eggs were exposed to higher incubation temperature (40.5°C) for 3 h on the 15th, 16th, and 17th days of incubation. The chicks obtained from each group were further subdivided and reared under different environmental conditions from the 15th to the 42nd day as normal [N; 25 ± 1 °C, 70% relative humidity (RH)] and heat exposed (HE; 35 ± 1 °C, 50% RH) resulting in four treatment groups (CN, CHE, TCN, and TCHE). The results revealed that HSP promoter activity was stronger in CHE, which had lesser methylation and higher gene expression. The activity of promoter region was lesser in TCHE birds that were thermally manipulated at the embryonic stage, thus reflecting their stress-free condition. This was confirmed by the lower level of mRNA expression of all the HSP genes. In conclusion, thermal conditioning during embryogenesis has a positive impact and improves chicken thermotolerance capacity in postnatal life.     

Expression of metabolic enzyme genes and heat-shock protein genes during embryonic development in diapause and non-diapause egg of multivoltine silkworm <Emphasis Type="Italic">Bombyx mori</Emphasis>

The expression of metabolic enzyme genes and heat-shock protein genes (Hsp) during early embryogenesis in diapause and non-diapause eggs of the silkworm Bombyx mori was quantified by semi-quantitative RT-PCR. The trehalase gene (Tre) was expressed in non-diapause eggs up-to nine days, while in diapause eggs was not up regulated. The glycogen phosphorylase gene (GPase) was expressed in non-diapause eggs, whereas in diapause eggs a high level was observed in early stage, but down regulated in later stage. The phosphofructokinase gene (PFK) and sorbitol dehyrogenase-2 gene (SDH-2) expression was fluctuated in non-diapause eggs, whereas in diapause eggs these were expressed only at early stage and not observed in later stage. The glucose-6-phosphate dehydrogenase gene (G6P-DH) in non-diapause eggs was highly expressed during the differentiation phase and decreased in the organogenesis phase. In contrast to this, expression in diapause eggs was of low level during differentiation phase and of high level observed in the organogenesis phase. In the tissues, PFK and SDH-2 were selectively expressed in cuticle and midgut, whereas Tre expression was high in midgut and ovary of larvae incubated at 15°C. The Hsp (20.4, 20.8, 40, 70, and 90) were expressed in both diapause and non-diapause eggs. Their expression was, however, selective in tissues with Hsp20.4 in midgut and ovary, Hsp40 in head, Hsp70 in cuticle and Hsp90 in ovary and head in high amounts at 15°C. These results suggest that the metabolic enzyme genes studied except Hsp play a major role during embryogenesis of diapause and non-diapause silkworm.     

Genome‐wide identification and characterization of HSP gene superfamily in whitefly (Bemisia tabaci) and expression profiling analysis under temperature stress

Heat shock proteins (HSP)are essential molecular chaperones that play important roles in the stress stimulation of insects.Bemisia tabaci,a phloem feeder and invasive species,can cause extensive crop damage through direct feeding and transmission of plant viruses.Here we employed comprehensive genomics approaches to identity HSP superfamily members in the Middle East Asia Minor 1 whitefly genome.In total,we identified 26 Hsp genes,including three Hsp90,17 Hsp70,one Hsp60 and five sHSP (small heat shock protein)genes.The HSP gene superfamily of whitefly is expanded compared with the other five insects surveyed here.The gene structures among the same families are relatively conserved.Meanwhile,the motif compositions and secondary structures of BtHsp proteins were predicted.In addition,quantitative polymerase chain reaction analysis showed that the expression patterns of BtHsp gene superfamily were diverse across different tissues of whiteflies.Most Hsp genes were induced or repressed by thermal stress (40℃)and cold treatment (4℃)in whitefly.Silencing the expression of BtHsp70-6 significantly decreased the survival rate of whitefly under 45℃.All the results showed the Hsps conferred thermo-tolerance or cold-tolerance to whiteflies that protect them from being affected by detrimental temperature conditions.Our observations highlighted the molecular evolutionary properties and the response mechanism to temperature assaults of Hsp genes in whitefly.     

Response of Heat-Shock Protein (HSP) Genes to Temperature and Salinity Stress in the Antarctic Psychrotrophic Bacterium Psychrobacter sp. G

Temperature and salinity fluctuations are two of the most important factors affecting the growth of polar bacteria. In an attempt to better understand the function of heat-shock proteins (HSPs) in the adaptive mechanisms of the Antarctic psychrotrophic bacterium Psychrobacter sp. G to such conditions, genes Hsp845, Hsp2538, Hsp2666, and Hsp2667 were cloned on the basis of the draft genome. The expression characteristics of these HSP genes under different stress conditions were analyzed by the qRT-PCR method. Expression of Hsp845 and Hsp2667 was inhibited significantly by low temperature (0 and 10 °C, respectively). There was no difference of expression when Hsp2538 and Hsp2666 were exposed to 0 °C but the expression of Hsp2666 was inhibited when exposed to 10 °C. Expression of Hsp2538 and Hsp2667 was not sensitive but expression of Hsp845 and Hsp2666 was increased at low salinity (0 and 15, respectively). Expression of the four HSP genes was enhanced at high salinity (90 and 120) and at high temperature independent of salinity. By contrast, low temperature had no significant effect independent of salinity.     

Expression of hsp70, hsp100 and ubiquitin in Aloe barbadensis Miller under direct heat stress and under temperature acclimation conditions

Key message

The study determined the tolerance of Aloe vera to high temperature, focusing on the expression of hsp70 , hsp100 and ubiquitin genes. These were highly expressed in plants acclimated at 35 °C prior to a heat shock of 45 °C.

Abstract

Aloe barbadensis Miller (Aloe vera), a CAM plant, was introduced into Chile in the semiarid IV and III Regions, which has summer diurnal temperature fluctuations of 25 to 40 °C and annual precipitation of 40 mm (dry years) to 170 mm (rainy years). The aim of this study was to investigate how Aloe vera responds to water and heat stress, focusing on the expression of heat shock genes (hsp70, hsp100) and ubiquitin, which not studied before in Aloe vera. The LT₅₀ of Aloe vera was determined as 53.2 °C. To study gene expression by semi-quantitative RT-PCR, primers were designed against conserved regions of these genes. Sequencing the cDNA fragments for hsp70 and ubiquitin showed a high identity, over 95 %, with the genes from cereals. The protein sequence of hsp70 deduced from the sequence of the cDNA encloses partial domains for binding ATP and the substrate. The protein sequence of ubiquitin deduced from the cDNA encloses a domain for interaction with the enzymes E2, UCH and CUE. The expression increased with temperature and water deficit. Hsp70 expression at 40–45 °C increased 50 % over the controls, while the expression increased by 150 % over the controls under a water deficit of 50 % FC. The expression of all three genes was also studied under 2 h of acclimation at 35 or 40 °C prior to a heat shock at 45 °C. Under these conditions, the plants showed greater expression of all genes than when they were subjected to direct heat stress.     

Effects of dietary L-carnitine and coenzyme Q10 at different supplemental ages on growth performance and some immune response in ascites-susceptible broilers

Abstract

Effects of dietary L-carnitine and coenzyme Q₁₀ (CoQ₁₀) at different supplemental ages on performance and some immune response were investigated in ascites-susceptible broilers. A 3 × 2 × 2 factorial design was used consisting of L-carnitine supplementation (0, 75, and 100 mg/kg), CoQ₁₀ supplementation (0 and 40 mg/kg) and different supplemental ages (from day 1 on and from day 10 on). A total of 480 one-day-old Arbor Acre male broiler chicks were randomly allocated to 12 groups, every group had five replicates, each with eight birds. The birds were fed a corn-soybean based diet for six weeks. From day 10 – 21, all the birds were exposed to a low ambient temperature (12 – 15°C) to increase the susceptibility to ascites. No significant effects were observed on growth performance by L-carnitine, CoQ₁₀ supplementation, and different supplemental ages. Packed cell volume was significantly decreased by L-carnitine supplementation alone, and ascites heart index and ascites mortality were decreased by L-carnitine, CoQ₁₀ supplementation alone, and L-carnitine + CoQ₁₀ supplementation together (p < 0.05). Heart index of broilers was significantly improved by L-carnitine, CoQ₁₀ supplementation alone during 0 – 3 week. Serum IgG content was improved by L-carnitine supplementation alone (p < 0.05), but lysozyme activity was increased by L-carnitine + CoQ₁₀ supplementation together (p < 0.05). A significant L-carnitine by supplemental age interaction was observed in lysozyme activity. L-carnitine supplementation alone had no effects on the peripheral blood lymphocyte (PBL) proliferation in response to concanavalin A (ConA) and lipopolysaccharide, but supplemental CoQ₁₀ alone and L-carnitine + CoQ₁₀ together decreased the PBL proliferation in response to ConA (p < 0.05). The present study suggested that L-carnitine + CoQ₁₀ supplementation together had positive effects on some immune response of ascites-susceptible broilers, which might benefit for the reduction of broilers' susceptibility to ascites.     

Expression of selected heat shock proteins after individually applied and combined drought and heat stress

Drought and heat stress are among the abiotic factors causing the most severe damage on plant crops. Their combination is quite common in dry and semi-dry regions worldwide and little is known about its effect on heat shock protein (HSP) profile in wheat plants. The expression of four HSP genes (Hsp 17.8, Hsp 26.3, Hsp 70 and Hsp 101b) in Triticum aestivum L. plants subjected to individually applied water deprivation or high temperature and their combination was monitored via one-step RT-PCR analysis. Changes in the expression levels of small HSPs (smHSPs), HSP70 and HSP100 were established also by SDS-PAGE. The combination of drought and heat induced HSP expression more effectively than the individually applied stresses. The induction of HSPs displayed greater rate in the drought-tolerant wheat variety Katya than in the drought-sensitive cv. Sadovo. The results obtained in wheat plants suggested that the effect of separately applied drought and heat shock cannot be extrapolated to their combination.     

Early age thermal manipulation on the performance and physiological response of broiler chickens under hot humid tropical climate

Initial brooding temperature is critical for post-hatch growth of broiler chickens. A study was conducted to investigate the early age thermal manipulation (EATM) on the performance and physiological responses broiler chickens under hot humid tropical climate. A total of 260 unsexed day-old Arbor-acre broiler chicks were assigned to five thermal treatments of brooding temperature regimens having 4 replicates of thirteen birds each. The heat treatments were: initial brooding temperature of 35 °C for the first 2 days, and then decreased subsequently, gradually to 22 °C at 21 d of age (CT), initial temperature of 35 °C, sustained for the first 4 days and then decreased gradually (conventionally) (FD), initial temperature of 35 °C for the first 7 days (SD), the birds in CT, but the brooding temperature was raised to 35 °C again for another 3 days from day 7 (SD3), initial brooding temperature of 35 °C for the first 10 days (TD). Data were collected on daily feed intake and weekly body weights. Blood samples were collected from 8 birds per treatment weekly for the determination of plasma uric acid, triglycerides, triiodothyronine (T₃) and creatinine kinase. Data obtained were laid out in a Completely Randomized Design (CRD). Results showed that the final weights of the birds in FD were higher (P < 0.05) than those of the other treatments at the finisher phase. Feed intake of the birds in FD was higher than those of SD3 and TD. FCR of broiler chickens in CT, SD, SD3 and TD was higher than that of FD. The rectal temperature, plasma MDA and blood glucose of the thermally challenged birds in FD was generally better (P < 0.05) than those of the other treatments. It was concluded that EATM can be used to improve performance and also protect broiler chickens from acute heat stress at market age.     

Myocardial expression of transforming growth factor beta family and endothelin-1 in the progression from heart failure to ascites in broilers with cold-induced pulmonary hypertension

We determined mRNA expression of genes of endothelin-1 (ET-1), and of the transforming growth factor beta ligands (TGFβ1, TGFβ2 and TGFβ3), their receptors (TβRI and TβRII) and their pseudoreceptor BAMBI in the heart of broilers raised under cold temperature conditions and affected by pulmonary hypertension. Gene expression was determined by RT-qPCR in right myocardial ventricle samples from 4-week-old chickens (n?=?48) raised either under normal (control) or cold temperature conditions (22?°C versus 14?°C). We do not find differences among healthy birds, birds with cardiac failure and ascitic birds in the mRNA levels of TGFβ2, TGFβ3 and BAMBI. In the control group, ET-1 mRNA level was increased in the ascitic birds as compared with healthy birds and birds with cardiac failure (p?<?0.05) whereas in the cold treated group, no increase was observed (p?>?0.05); yet, ascitic birds in the cold group showed lower mean than ascitic birds in the control group (p?<?0.05). TβRII mRNA expression was higher in ascitic than in healthy birds (p?<?0.05) in both control and cold treated groups; however, in the ascitic birds of the cold treated group TβRII expression was lower than in ascitic birds from the control group (p?<?0.05). Thus, the higher ET-1 and TβRII levels observed in ascitic birds seem to be attenuated by cold.     

CHANGES IN CELLULAR PROTEINS DUE TO ENVIRONMENTAL NON-IONIZING RADIATION. I. HEAT-SHOCK PROTEINS

This paper describes the effect of weak microwave fields on the amounts of heat-shock proteins in cell cultures at various temperatures.

The field was generated by signal simulation of the Global System for Mobile communications (GSM) of 960 Mhz, used in portable phones. Transformed human epithelial amnion (AMA) cells, growing on glass coverslips, were exposed in a transverse electromagnetic (TEM) cell to a microwave field, generating a specific absorption rate (SAR) of 2.1 mW.kg^?1 in the cells. Exposure temperatures were 35, 37, and 40 ± 0.1°C, respectively, and the exposure time was 20 min.

The heat-shock proteins Hsp-70 and Hsp-27 were detected by immuno-fluorescence. Higher amounts of Hsp-70 were present in the cells exposed at 35 and 37°C than in the sham-exposed cells.

These effects can be considered to be athermal, since the field strength was much lower than the safety standard for absence of heat generation by microwave fields.

There was no significant response in the case of Hsp-27.     

Effects of source and level of magnesium on catalase activity and its gene expression in livers of broiler chickens

Abstract

The effects of dietary supplemental magnesium oxide (MgO), magnesium-L-aspartate (MgAsp) and monomagnesium-di-L-aspartate (MgdiAsp) on hepatic catalase (CAT) activity and its mRNA expression were investigated. A total of 360 one-day-old male Abor Acre broiler chickens were allocated to ten treatments, i.e. control plus 9 treatments from 3×3 factorial arrangement (Mg source, Mg level), each treatment with six replicates of 6 chickens. The birds were fed with the basal diet alone or supplemented with magnesium (Mg) at 0.9, 1.8, 2.7 g/kg of the diet from MgO, MgAsp or MgdiAsp. Results showed that hepatic Mg concentration increased quadratically as MgO or MgAsp supplementation increased (p < 0.01). Hepatic CAT activity increased linearly in birds fed with MgAsp or MgdiAsp (p < 0.01) and quadratically in birds fed with MgO (p < 0.05) as dietary Mg supplementation level increased. Hepatic CAT mRNA was linearly correlated with the dietary Mg supplementation level (p < 0.01). There were positive correlations among hepatic CAT activity, its mRNA expression level and hepatic Mg concentration (p < 0.01). No effect of Mg²⁺ on the purified CAT activity was detected in vitro enzymatic reaction system (p > 0.05). Supplemental MgAsp or MgdiAsp was more efficient to increase hepatic Mg concentrations, enhance hepatic CAT activity and its mRNA expression than MgO (p < 0.01). It can be concluded that dietary Mg supplementation could increase hepatic Mg concentration, enhance CAT mRNA expression and consequently enhance CAT activity, and the organic Mg (MgAsp or MgdiAsp) is much more efficient than the inorganic form (MgO).     

Hsp-72, a candidate prognostic indicator of heatstroke

Exposure of rats to environmental heat enhances the expression of heat shock protein-72 (Hsp-72) in most of their organs proportionally to heat stress severity. Pre-induction or over-expression of Hsp-72 prevents organ damage and lethality, suggesting that heat shock proteins (Hsps) may have a pathogenic role in this condition. We investigated the expression profile of Hsps in baboons subjected to environmental heat stress until the core temperature attained 42.5°C (moderate heatstroke) or occurrence of hypotension associated with core temperature ≥43.5°C (severe heatstroke). Western blot analysis demonstrated a differential induction of Hsp-72 among organs of heat-stressed animals with the highest induction in the liver and the lowest in lung. Hsp-60 and Hsc-70 expression was similar between control and heat-stressed animals. ELISA studies indicated a marked release of Hsp-72 into the circulation of baboons with severe heatstroke with a peak at 24 h post-heatstroke onset and remained sustained up to 72 h. Hsp-72 release was not associated with core temperature or systolic blood pressure, but correlated with markers of liver, myocardium, and skeletal muscle tissue necrosis. Non-survivors displayed significantly higher Hsp-72 levels than survivors. No Hsp-60 was detected in the circulation. These findings add further evidence that increased expression of Hsp-72 may be an important component of the host response to severe heatstroke. They also suggest that extracellular Hsp-72 is a marker of multiple organs tissue damage. Whether extracellular Hsp-72 plays a role in the host immune response to heat stress merits further studies.     

Immunopathological effect of the mycotoxins cyclopiazonic acid and T-2 toxin on broiler chicken

Forty, newly hatched, unsexed broiler chicks were fed diets containing 10 ppm cyclopiazonic acid (CPA) and 1 ppm T-2 toxin (T₂) either individually or in combination for 28 days to study the immunopathological effects. Lymphoid organs revealed lymphocytolysis and lymphoid depletion in all toxin fed birds. Thymic and splenic CD⁺₄ and CD⁺₈ lymphocytes decreased significantly (p < 0.01) in toxin fed birds when compared to the control. Thymic CD⁺₈ lymphocytes of T₂ and CPA-T₂ showed significant (p < 0.01) decrease from that of CPA and control groups. Splenic CD⁺₄ and CD⁺₈ lymphocytes showed significant (p < 0.01) decrease in CPA and CPA-T₂ fed groups when compared to the control. The T₂ group did not differ significantly from that of control. The stimulation index (SI) of splenocytes to concavalin A revealed significant (p < 0.01) decrease in all toxin fed birds. Significant (p < 0.01) decrease were observed for the haemagglutination inhibition (HI) titres to Newcastle disease virus vaccine F strain (NDV) of birds fed CPA, T₂ and in combination. Significant (p < 0.01) interaction was found for lymphocyte subsets, SI and HI titres to NDV. The study indicated the immunosuppressive effect of these toxins either alone or in combination in broiler chicks.Forms part of M.V.Sc. thesis of the first author approved by the Tamil Nadu Veterinary and Animal Sciences University, Chennai 600 051, India.     

Melatonin supplementation can ameliorate the detrimental effects of heat stress on performance and carcass traits of Japanese quail

Melatonin, the major pineal hormone, modulates growth in poultry by influencing hormones involved in growth. We investigated the effects of dietary melatonin supplementation on performance, carcass characteristics, and excretion of nitrogen and some minerals in broiler Japanese quails (Coturnix coturnix japonica) exposed to high-ambient-temperature stress (34°C). One hundred twenty Japanese quails (10 d old) were randomly assigned to 4 treatment groups, 3 replicates of 10 birds each. The birds were kept in either an environment-controlled room at a constant 22°C or were kept at 22°C for 16 h/d and at 34°C for 8 h/d (9:00 am to 5:00 pm). At both temperatures birds were fed either a basal diet or the basal diet supplemented with 40 mg of melatonin per kilogram of diet. The experiment lasted for 32 d. Melatonin improved feed efficiency in both temperatures groups compared with their corresponding controls. Although feed intake was similar in all groups, the improvement in feed efficiency was more noticable in melatonin-fed quails kept at high temperature (p<0.01). Supplemental melatonin significantly increased live weight gain and carcass characteristics under stress conditions (p<0.01) but did not show the same effect at thermoneutral conditions (p>0.05). Heat exposure increased excretion of N, Ca, P, Zn, Fe, and Cr and decreased retention rates for them. Dietary melatonin supplementation returned these values to normal (p<0.01). No interactions between melatonin and temperature were found in the parameters measured. The results of the study show that melatonin supplementation attenuated the retardation in performance as well as the excretion of minerals caused by heat stress in broiler quails. Our data suggest that melatonin might offer protection against heat-stress-related depression in the performance of broiler quails.     

<Emphasis Type="Italic">Treponema denticola</Emphasis> alters cell vitality and induces HO-1 and Hsp70 expression in porcine aortic endothelial cells

Treponema denticola is an oral spirochete that is associated with periodontal disease and detected occasionally in extraoral lesions associated with systemic disorders such as cardiovascular diseases. The effect of specific bacterial products from oral treponemes on endothelium is poorly investigated. This study analyzed the ability of components of the outer membrane of T. denticola (OMT) to induce apoptosis and heat shock proteins (HO-1 and Hsp70) in porcine aortic endothelial cells (pAECs), compared with results obtained with classical pro-inflammatory lipopolysaccharide (LPS) treatment. Cellular apoptosis was detected when pAECs were treated with either OMT or LPS, suggesting that OMT can damage endothelium integrity by reducing endothelial cell vitality. Stimulation with OMT, similarly to LPS response, increased HO-1 and Hsp-70 protein expression in a time-dependent manner, correlating with a rise in HO-1 and Hsp-70 mRNA. Collectively, these results support the hypothesis that T. denticola alters endothelial cell function. Moreover, our in vitro experiments represent a preliminary investigation to further in vivo study using a pig model to elucidate how T. denticola leaves the initial endodontic site and participates in the development of several systemic diseases.