Hsp70 Gene Expression Research Articles

Interactive effects of methionine source and carnitine supplementation on growth performance, immunity, antioxidant status, and HSP70 gene expression in broilers reared under heat stress conditions

Interactive effects of methionine source and carnitine supplementation on growth performance, immunity, antioxidant status, and HSP70 gene expression in broilers reared under heat stress conditions

Biological responses of stellate sturgeon fingerlings (Acipenser stellatus) immersed in HSP inducer to salinity changes

Changes in salinity is a stressful and energy-consuming process in fish which give rise to mortalities, especially in fish fingerlings that are more sensitive during the early stages of their life. In the present study, the effects of three salinities, 3‰ (downstream of river), 8‰ (estuarine), and 13‰ (the maximum salinity in the Caspian Sea), on HSP70 gene expression, cortisol level, immune response (lysozyme, complement C3, IgM), and antioxidant enzyme activities (SOD, CAT, T-AOC) of the stellate sturgeon fingerlings in the presence of HSP inducer compound (TEX-OE®) were evaluated. Our results showed that levels of plasma cortisol and heat shock protein (HSP70) in Acipenser stellatus fingerlings increased due to salinity changes. In the presence of the HSP inducer, HSP70 expression in both gill and liver was significantly increased, whereas cortisol level was notably decreased. Exposure to salinity changes resulted in an increase in antioxidant defense activities (SOD, CAT, and T-AOC) and immune response (lysozyme, IgM, and C3) in the presence of an HSP inducer. In conclusion, an HSP-inducing compounds can have a positive effect in strengthening the immunity and antioxidant system of sturgeon fingerlings by increasing the expression of the HSP70 gene against salinity fluctuations and generally increase the body's physiological tolerance.

Mycorrhizal Fungal Effects on Plant Growth, Osmolytes, and CsHsp70s and CsPIPs Expression in Leaves of Cucumber under a Short-Term Heat Stress.

Arbuscular mycorrhizal (AM) fungi enhance plant stress tolerance, but it is unclear whether AM fungi affect heat tolerance in cucumbers. This study aimed to analyze how an AM fungus, Diversispora versiformis, affected growth, chlorophyll, five osmolytes, and plasma membrane intrinsic proteins (PIPs) and heat shock protein 70 (Hsp70) gene expression in cucumber leaves after a short-term (80 h) heat stress. Heat treatment significantly reduced root AM fungal colonization rate (0.26 folds). Heat treatment also distinctly suppressed plant height, stem diameter, and biomass, whereas AM fungal inoculation improved these growth variables as well as the chlorophyll index, with the benefit being more obvious under heat than under no-heat stress conditions. Heat treatment triggered differential changes in osmolytes (sucrose, fructose, and betaine) of inoculated and uninoculated cucumbers, whereas inoculation with AM fungus significantly raised leaf sucrose, fructose, glucose, betaine, and proline levels when compared to non-AM fungal inoculation. Heat treatment increased the expression of two (CsPIP1;6 and CsPIP2;1) of eight CsPIPs in inoculated and uninoculated plants, whereas AM fungal inoculation up-regulated the expression of CsPIP1;6, CsPIP2;1, and CsPIP2;6 under heat stress conditions. Hsp70s expressed differently in inoculated and uninoculated plants under heat versus no-heat stress, with 6 of 11 CsHsp70s down-regulated in inoculated plants. Under heat stress conditions, AM fungus only up-regulated CsHsp70-8 expression in 11 Hsp70s, while another eight CsHsp70s were down-regulated. Heat treatment and AM fungal inoculation both increased the expression of CsHsp70-8 and CsPIP1;6. It was concluded that AM fungus-inoculated cucumbers have high levels of growth, chlorophyll, and osmolytes under heat stress and do not require high CsPIPs and CsHsp70s expression to tolerate a short-term heat treatment.

The first reporting of prevalence Vibrio species and expression of HSP genes in rayed pearl oyster (Pinctada radiata) under thermal conditions.

The main goal of the present study was to evaluate the influence of thermal exposure on Vibrio population and HSP genes expression (HSP 90, HSP70, and HSP20) in rayed pearl oyster (P. radiata). To this end, the oysters were reared for 30 days at temperatures of 22°C (control), 25°C, 27°C, and 29°C. The results showed that five dominate Vibrio strains including Vibrio hepatarius, V. harveyi, V. alginolyticus, V. parahaemolyticus, and V. rotiferianus were identified. The highest population of V. parahaemolyticus, V. alginolyticus, and V. harveyi, was found in 29οC group. According to real-time PCR, mantle exhibited the highest expression levels of HSP20, HSP70, and HSP90 genes. A higher level of HSP20 expression was observed at high temperatures (25°C, 27°C, and 29°C) in the gonad and mantle compared to the control group (22°C) while decrease in HSP90 expression level was recorded in 25°C, 27°C, and 29°C groups. HSP20 expression level in adductor muscle was remarkably down-regulated in 27°C and 29°C groups. In this tissue, HSP70 was detected at highest levels in the 29οC group. In mantle, HSP90 gene expression was lowest at 22°C water temperature. Several Vibrio strains have been identified from pearl Gulf oyster that haven't been previously reported. The identification of dominant Vibrio species is essential for epidemiological management strategies to control and prevent Vibrio outbreaks in pearl oyster farms. The expression pattern of HSP genes differs in rayed pearl oyster tissues due to differences in their thermal tolerance capability and physiological and biological characteristics. The present study provides useful molecular information for the ecological adaptation of rayed pearl oysters after exposure to different temperature levels.

Biomarkers related to gas embolism: Gas score, pathology, and gene expression in a gas bubble disease model.

Fish exposed to water supersaturated with dissolved gas experience gas embolism similar to decompression sickness (DCS), known as gas bubble disease (GBD) in fish. GBD has been postulated as an alternative to traditional mammals' models on DCS. Gas embolism can cause mechanical and biochemical damage, generating pathophysiological responses. Increased expression of biomarkers of cell damage such as the heat shock protein (HSP) family, endothelin 1 (ET-1) or intercellular adhesion molecule 1 (ICAM-1) has been observed, being a possible target for further studies of gas embolism. The GBD model consisted of exposing fish to supersaturation in water with approximately 170% total dissolved gas (TDG) for 18 hours, producing severe gas embolism. This diagnosis was confirmed by a complete histopathological exam and the gas score method. HSP70 showed a statistically significant upregulation compared to the control in all the studied organs (p <0.02). Gills and heart showed upregulation of HSP90 with statistical significance (p = 0.015 and p = 0.02, respectively). In addition, HSP70 gene expression in gills was positively correlated with gas score (p = 0.033). These results suggest that gas embolism modify the expression of different biomarkers, with HSP70 being shown as a strong marker of this process. Furthermore, gas score is a useful tool to study the abundance of gas bubbles, although individual variability always remains present. These results support the validity of the GBD model in fish to study gas embolism in diseases such as DCS.

Sub-chronic exposure of Oreochromis niloticus to environmentally relevant concentrations of smaller microplastics: Accumulation and toxico-physiological responses

This study assesses the accumulation and toxic effects of environmentally relevant concentrations (0.01, 0.1 and 1 mg/L) of polystyrene MPs (1 µm) in Oreochromis niloticus (Nile tilapia) for 14 days. The results showed that 1 µm PS-MPs accumulated in the intestine, gills, liver, spleen, muscle, gonad and brain. RBC, Hb and HCT showed a significant decline, while WBC and PLT showed a significant increase after the exposure. Glucose, total protein, A/G ratio, SGOT, SGPT and ALP showed significant increments in 0.1 and 1 mg/L of PS-MPs treated groups. The increase in cortisol level and upregulation of HSP70 gene expression in response to MPs exposure indicate MPs-mediated stress in tilapia. MPs-induced oxidative stress is evident from reduced SOD activity, increased MDA levels and upregulated P53 gene expression. The immune response was enhanced by inducing respiratory burst activity, MPO activity and serum TNF-α and IgM levels. MPs exposure also led to down-regulation of CYP1A gene and decreased AChE activity, GNRH and vitellogenin levels, indicating the toxicity of MPs on the cellular detoxification mechanism, nervous and reproductive systems. The present study highlights the tissue accumulation of PS-MP and its effects on hematological, biochemical, immunological and physiological responses in tilapia with low environmentally relevant concentrations.

Effect of essential oils blend inclusion on performance and HSP70 gene expression in broilers chickens under chronic heat stress

The objective of this study was to evaluate the effects of the inclusion of a blend of essential oils (capsaicin, carvacrol, cinnamaldehyde and eugenol) in broiler submitted to thermal stress. Performance parameters and gene expression of heat shock protein (HSP70) were evaluated in the liver and jejunum. Regarding performance, animals subjected to heat stress had lower feed intake and weight gain and higher feed conversion. The inclusion of 3% of the blend increased the HSP70 gene expression in jejunum and liver of 42 days old broiler chickens.

Sildenafil citrate ameliorates the adverse effects of cryopreservation on sperm quality in asthenozoospermic men

In this study, the effects of Sildenafil Citrate on the sperm quality during cryopreservation in the asthenozoospermic patients were investigated for the first time. Thirty semen samples were collected from asthenozoospermic patients and each sample was divided into 3 groups: Control (fresh), Freeze and Freeze + Sildenafil. In each groups the sperm parameters, DNA fragmentation, acrosome integrity, protamine deficiency, mitochondrial membrane potential, plasma membrane integrity, the expression of Bcl-2 and HSP70 genes, as well as the level of Tumor necrosis factor-alpha, Malondialdehyde and antioxidants (Catalase, Glutathione, and Superoxide dismutase) in sperm were assessed. Data were analyzed statistically using Repeated Measure Analysis. The level of Malondialdehyde and Tumor necrosis factor-alpha, morphological abnormalities, DNA fragmentation, protamine deficiency and the expression of Bcl-2 and HSP70 genes increased significantly in the Freeze group compared to the Control, while the level of sperm parameters and antioxidants, plasma membrane integrity, mitochondrial membrane potential and acrosomal integrity significantly decreased. In the Freeze + Sildenafil group, compared to the Freeze group, all the mentioned parameters were significantly reversed except for the acrosomal integrity (decreased even more) and the expression of Bcl-2 (increased even more) and HSP70 genes (with no change). Although adding Sildenafil to the freezing medium decreased the adverse effects of freezing on the sperm of asthenozoospermic patients and improved sperm quality, but it also caused premature acrosome reaction. Therefore, we suggest the consumption of Sildenafil along with another antioxidant, to benefit from the favorable effects of Sildenafil as well as to maintain the integrity of the sperm acrosome.

Construction of feature analysis model for demeanor evidence investigation based on data mining algorithm

Demeanor evidence refers to the authoritative opinions of experts using analytical instruments to collect and interpret the formation of external physiological symptoms of the entire human body. With the development of modern technology, the observation of demeanor has entered a new stage of multimodal quantification. The quantifiable results of demeanor observation with the help of scientific and technological instruments play an important role in judicial work such as search, interrogation, and evidence review. The use of man-made reasoning in the field of legal settling has become increasingly broad. Specialists are giving increasingly more consideration to the securing of modular proof. This paper proposes the development of a component examination model for modular proof examination applications. The technique for this paper is to apply the guileless Bayes strategy, propose a superior information mining calculation, and lay out a model for proof observation and examination. The function of these methods is to systematically explore the basic theoretical issues of demeanor evidence based on the status quo of judicial application of demeanor evidence. Through the prediction of individual demeanor based on data mining algorithm, the evidence analysis model is designed, the neurobiological experiment is carried out, and the demeanor evidence animal stress model is constructed to verify the scientific basis of multidemeanor evidence observation. The experimental results showed that after repeated stimulation of SD rats, the maximum changes in the expression of HSP70 gene and SAA gene were 10.77 and 14.1 respectively, reflecting the high reliability of demeanor evidence biological experiments. The model can improve the accuracy of evidence use, and the correct use of demeanor evidence can realize the true litigation value of intelligent justice and the concept of human rights protection, and promote the construction of intelligent justice.

Exposure to a 900 MHz electromagnetic field induces a response of the honey bee organism on the level of enzyme activity and the expression of stress-related genes.

There are many artificial sources of radiofrequency electromagnetic field (RF-EMF) in the environment, with a value between 100 MHz and 6 GHz. The most frequently used signal is with a frequency of around 900 MHz. The direction of these changes positively impacts the quality of life, enabling easy communication from almost anywhere in the world. All living organisms in the world feel the effects of the electromagnetic field on them. The observations regarding the influence of a RF-EMF on honey bees, describing the general impact of RF-EMF on the colony and/or behavior of individual bees, such as reduction in the number of individuals in colonies, extended homing flight duration, decrease in breeding efficiency, changes in flight direction (movement of bees toward the areas affected by RF-EMF), increase in the intensity and frequency of sounds characteristic for those announcing the impending danger. In this work, we describe the changes in the levels of some of the stress-related markers in honey bees exposed to varying intensities and duration of RF-EMF. One-day-old honeybee worker bees were used for the study. The bees were randomly assigned to 9 experimental groups which were exposed to the following 900 MHz EMF intensities: 12 V/m, 28 V/m, and 61 V/m for 15 min, 1 h and 3 h. The control group was not exposed to the RF-EMF. Each experimental group consisted of 10 cages in which were 100 bees. Then, hemolymph was collected from the bees, in which the activity was assessed AST, ALT, ALP, GGTP, and level of nonenzymatic antioxidants albumin, creatinine, uric acid, and urea. Bees were also collected for the analysis of rps5, ppo, hsp10, hsp70, hsp90, and vitellogenin gene expression. Our study shows that exposure to a 900 MHz electromagnetic field induces a response in the honey bees that can be detected in the level of enzyme activity and the expression of stress-related genes. The response is similar to the one previously described as a result of exposition to UVB irradiation and most likely cannot be attributed to increased temperature.

Transcriptome analysis revealed the mechanism of Luciobarbus capito (L. capito) adapting high salinity: Antioxidant capacity, heat shock proteins, immunity

Salinity has a significant influence on the physiology of freshwater aquatic organisms. However, there are few studies on the hematology and immunology of freshwater fish under high salinity. In the current study, we aimed to analyze the adaptive effect of salt stress on L. capito spleen immune function and hematology using transcriptomic analysis. We replicated a L. capito acute salinity stress model, and collected blood and spleens from freshwater and saltwater fish. It was found that salinity affected significantly the numbers of leukocytes, lymphocytes, neutrophils, and red blood cells, as well as the content of haemoglobin. Salt treatment resulted in a significant increase in the expression of HSP70, HSP90, CAT, SOD, and GPX1 genes in L. capito spleens. Transcriptomic analysis revealed a total of 546 differentially expressed genes (DEGs) in spleens, including 224 up-regulated DEGs and 322 down-regulated DEGs. In addition, GO enrichment analysis revealed immune system process, multicellular organismal process, and biological regulation of genes with the most differences in biological processes. KEGG enrichment analysis showed that the regulation of lipolysis in adipocyte, FoxO signaling pathway, Hematopoietic cell lineage signaling pathway, and HIF-1 signaling pathway were significantly enriched. L. capito adapted oxidative to high salinity through FoxO signaling pathway and immune to high salinity through Hematopoietic cell lineage signaling pathway. At the same time, we selected 10 DEGs for qRT-PCR detection, and the results showed that the qRT-PCR results were consistent with our RNA-Seq results, indicating that transcriptome sequencing was accurate and reliable. In conclusion, our results demonstrated that the improvement of antioxidant capacity, heat shock protein and immunity are involved in the molecular mechanism of L. capito adapting to high salinity. Our findings provided a rationale for further study on high salinity adaptation and related enrichment pathways.

BuOH fraction of Salix Babylonica L. extract increases pancreatic beta-cell tumor death at lower doses without harming their function

Salix babylonica L. is a species of the willow tree. Insulinoma is a tumor originating from pancreatic beta cells. This study aims to research the effect of different fractions of Salix babylonica L. leaf extract on INS-1 cells for treating pancreatic tumors. Cell death occurred at lower doses in the EtOAc fraction. The cells are functional in the BuOH fraction but not in EtOAc and H2O fractions. The EtOAc fraction has a higher percentage of necrosis and ROS. INS1, INS2, and AKT gene expressions in the H2O fraction, GLUT2, IR, HSP70 gene expressions, and WNT4 protein levels increased in the BuOH fraction. HSP90 gene expression, Beta-actin, GAPDH, insulin, HSP70, HSP90, HSF1, Beta-Catenin, and WNT7A protein levels were decreased, while IR immunolabelling intensity increased in both fractions. Ca+2, K+, Na+, and CA-19-9 in the cell, Ca+2 and K+ in secretion increased. The secondary metabolites in the EtOAc fraction cause more damage in INS-1 cells. Since the water fraction also causes the cells to die in high doses, cell function is damaged. The secondary metabolites in the BuOH fraction kill INS-1 cells with less damage. This makes the BuOH fraction of Salix babylonica L. more valuable.

Seasonal Effect of THI on HSP70 Gene Expression Pattern in Deccani Sheep

The region of Satara district experiences summer from March to May, with maximum temperature ranging from 35°C to 43°C and the warmest month is May (43°C) whereas, the monsoon lasts from June to September with moderate to high precipitation during which temperature ranges from 28°C to 32°C along with 80% to 90% relative humidity. The present study was designed to evaluate the effect of THI on the HSP70 gene expression pattern during summer and rainy seasons in Deccani sheep at LFC, KNP College of Veterinary Science, Shirwal, Dist. Satara (Maharashtra). The meteorological variables like temperature and relative humidity were recorded for calculating THI and whole blood samples (06 ml each) of eight non-pregnant, non-lactating and apparently healthy Deccani sheep were collected for HSP70 gene expression pattern during peak summer (month of May) and peak rainy (month of August) seasons in the year 2019. The significantly (P less than 0.01) higher THI was recorded during summer than rainy season. The relative mRNA expression of the HSP70 gene during the peak summer season was found 1.79 folds more as compared to peak rainy season during the present study.

Eyestalk ablation and acclimation temperatures affect juveniles Penaeus vannamei thermo-tolerance: molecular biomarkers of cell protection oxidative stress, and compensatory mechanisms

This study investigated the effect of acclimation temperatures and eyestalk ablation (EA) on the thermal tolerance of juvenile Penaeus vannamei. In each case, the relative expression of genes involved in cell protection (hsp70 and hsp90), oxidative stress (cMnSOD and GPx), and anaerobic metabolism (hif1a) was assessed. For this purpose, shrimp were acclimated to 20, 26, and 32°C for 21 days. After acclimation, the critical thermal maxima (CTmax) was determined in non-eyestalk ablated, unilaterally, and bilaterally eyestalk ablated organisms. An effect of acclimation temperatures on CTmax values was observed, with shrimp acclimated at 32°C having the highest rates. Likewise, EA resulted in lower thermal tolerance to CTmax in organisms acclimated at 20 and 26°C. The shrimp's protective and cellular repair responses were evidenced by increased hsp70 and hsp90 gene expression after CTmax and were intensified by the EA. In contrast, the results showed that cMnSOD was very sensitive to CTmax, and its expression was intensified with EA, while for GPx, there was an increase in the relative gene expression, mainly in shrimp acclimated at 20°C. In the case of hif1a, overexpression was observed at the acclimation temperature of 26°C, showing the activation of compensatory mechanisms such as anaerobic metabolism. EA caused a significant molecular response during CTmax of molecular biomarkers involved in heat stress response, oxidative stress, and compensatory mechanisms.

The combined effects of polyethylene microplastics and benzoanthracene on Manila clam Ruditapes philippinarum

Microplastic (MP) toxicity has recently been explored in various marine species. Along with the toxicity of plastics polymer itself, additional substances or pollutants that are absorbed onto it may also be harmful. In the present study, we investigated the combined impacts of polyethylene microplastics (PE MPs) and an organic pollutant (Benzo(a)anthracene, BaA) on Manila clam Ruditapes philippinarum during a one-week exposure. Two PE MPs concentrations (26 μg L−1 and 260 μg L−1) and one BaA concentration (3 μg L−1) were tested. The clams were exposed to BaA and PE MPs either alone or in combination. BaA and PE MPs were incubated before the combined exposure. The biological effects of PE MPs and BaA on the clams were evaluated by considering several assays such as feeding rate, anti-oxidant enzyme activities, and the expression levels of stress-related genes. The feeding rate significantly decreased in individual PE MPs and individual BaA groups while it remained unchanged in combined groups. Superoxide dismutase (SOD) was the most affected among the biochemical parameters. Malondialdehyde (MDA), and glutathione peroxidase (GPx) activities were slightly affected, whereas no changes were observed in glutathione s-transferase (GST) activities. CYP1A1, CYP3A4, and HSP70 gene expressions displayed slightly significant changes. Considering all stressor groups, high PE MPs exposure (260 μg L−1 PE MPs) more effectively altered the biological parameters in the clams compared to individual low PE MPs and BaA exposure, and their combination. The results also indicated the negligible vector role of PE MPs to transport BaA into the clam tissues.

HSP70 Gene Expression Pattern in Peripheral Blood Mononuclear Cells during Different Seasons in Pandharpuri Buffaloes

Background: Heat shock protein is one of the most abundant and best characterized heat shock protein family that consists of highly conserved stress proteins. Amongst these heat shock protein family, HSP70 is expressed in response to stress and plays crucial roles in environmental stress tolerance and adaptation. Thermal stress alters the normal body homeostasis and causes severe detrimental effects on production and productivity of animals and impairs growth, immune status of animal. Therefore, the present study is proposed to study HSP70 gene expression pattern in peripheral blood mononuclear cells. Methods: The present study was conducted to study the relative mRNA expression pattern of HSP70 gene in peripheral blood mononuclear cells (PBMCs) during different seasons in Padharpuri buffaloes. 10 Apparently healthy, non - pregnant Pandharpuri buffaloes (above 2 years of age) were taken for study. Blood samples were collected thrice i.e. once in winter season, once in summer season and once in rainy season. Real-time polymerase chain reaction was applied to investigate mRNA expression of HSP70 gene during different seasons. Specificity of the desired products was documented using analysis of the melting temperature and high resolution gel electrophoresis to verify that the transcripts are of the exact molecular size predicted. Result: To investigate the variation in relative mRNA expression profile of HSP70 gene, Glyceraldehyde 3-phosphate dehydrogenase (GAPDH gene) was used as a housekeeping gene. The relative expression values of HSP70 during summer and rainy seasons were found statistically significant in comparison to winter. These results suggest that HSP70 gene expression varies with season and this variation may play an imperative role in conferring thermo tolerance against heat stress during different seasons.

Dietary Chlorella vulgaris effectively alleviates oxidative stress, immunosuppression, and enhances the resistance to Streptococcus agalactiae infection in cadmium-intoxicated Nile tilapia fingerlings

Aquatic pollutants, including cadmium (Cd), cause oxidative stress on aquatic animals. The use of probiotics, including microalgae as a feed additive to alleviate the toxic impacts of heavy metals, is a much more interesting point. Hence, the current study investigated the oxidative stress and immunosuppression in Nile tilapia (Oreochromis niloticus) fingerlings caused by Cd toxicity as well as the preventive function of dietary Chlorella vulgaris against Cd toxicity. Accordingly, fish were fed on 0.0 (control), 5, and 15 g/kg diet of Chlorella up to satiation thrice a day, along with being exposed to 0.0 or 2.5 mg Cd/L for 60 days. Following the experimental procedure, fish from each group were intraperitoneally injected with Streptococcus agalactiae, and their survivability was observed for further ten days. Chlorella-supplemented diets meaningfully (P < 0.05) boosted the antioxidative capability of fish, which was evidenced by higher activities of hepatic superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), and glutathione-S-transferase (GST) as well as higher levels of reduced glutathione (GSH) along with significant reductions in hepatic malondialdehyde levels. Moreover, the innate immunity indices [phagocytic activity (PA), respiratory burst activity (RBA), and alternative complement activity (ACH50)] were significantly higher in Chlorella-fed fish, particularly in the group of 15 g/kg diet. Additionally, serum of Chlorella-fed fish showed potent bactericidal activities against S. agalactiae, particularly at the treatment of a 15 g/kg diet. Feeding Chlorella diets to Nile tilapia fingerlings upregulated SOD, CAT, and GPx genes expression alongside the down-regulation of IL-1β, IL-8, IL-10, TNF-α, and HSP70 genes expression. Conversely, Cd toxicity caused oxidative stress and suppressed the fish's innate immunity with upregulation of the expression of IL-1β, IL-8, IL-10, TNF-α, and HSP70 genes. Feeding Cd-exposed fish on Chlorella-containing diets attenuated these adverse effects. The current research revealed that supplementing feeds with the treatment of 15 g/kg diet of C. vulgaris supports the antioxidant-immune responses and alleviates the Cd toxicity effects on Nile tilapia fingerlings.

Estimating the intra-puparial period of Hydrotaea spinigera (Stein,1910) (Diptera: Muscidae) with morphological and gene expression changes

Hydrotaea spinigera (Stein, 1910) (Diptera: Muscidae) is a forensically important sarcosaprophagous species widely distributed throughout the Oriental and Australasian regions. At the advanced decomposition stage or the skeletonize stage, the immature stages of H. spinigera, especially the pupae, can still be found in large quantities and could be used as important indicators to estimate the minimum postmortem interval (PMImin). However, there have been no studies on the intra-puparial period of this species. Herein, we studied morphological and differential gene expression changes during the intra-puparial development of H. spinigera, aiming to accurately estimate the intra-puparial age of H. spinigera. The intra-puparial morphological changes of H. spinigera were observed at seven constant temperatures ranging from 16 °C to 34 °C and divided into 12 sub-stages. Structures that could be used to estimate the intra-puparial age, such as compound eyes, mouthparts, antennae, thorax, legs, wings, and abdomen, were observed in detail, and the developmental process of each structure was divided into 5 to 10 stages. The time range of each sub-stage, or when a structure appeared, was recorded. For the gene expression section, the most suitable reference genes were screened by geNorm, NormFinder, BestKeeper, and ΔCt methods. Based on the selected reference genes, real-time quantitative PCR (RT qPCR) was used to detect the expression changes of the white and hsp90 genes with developmental time at 19 °C, 25 °C, and 31 °C. Results showed that the trend of hsp90 gene expression under different temperatures was not consistent, while white genes exhibited regular changes during development, and could thus be used for age estimation of H. spinigera. This study provides an important basis for forensic entomology to use morphological and differential gene expression for estimating the age of H. spinigera during the intra-puparial period. Moreover, the combination of the two methods can produce a more accurate minimum postmortem interval (PMImin) estimate compared to when each method is used separately.

The impact of ascidian biofouling on the farmed Mediterranean mussel Mytilus galloprovincialis physiology and welfare, revealed by stress biomarkers

In biofouling communities, ascidians are among the most damaging species, presenting severe threats, such as depressed growth rates and decreased chances of lower survival, to shellfish aquaculture. However, little is known concerning the fouled shellfish physiology. In an effort to obtain information for the magnitude of stress caused by ascidians to farmed Mytilus galloprovincialis, five seasonal samplings took place in a mussel aquaculture farm suffering from ascidian biofoulants, in Vistonicos Bay, Greece. The dominant ascidian species were recorded and several stress biomarkers, including Hsp gene expression at both mRNA and protein levels, as well as MAPKs levels, and enzymatic activities of intermediate metabolism were examined. Almost all investigated biomarkers revealed elevated stress levels in fouled mussels compared to non-fouled. This enhanced physiological stress seems to be season-independent and can be attributed to the oxidative stress and/or feed deprivation caused by ascidian biofouling, thus illuminating the biological impact of this phenomenon.

Conferring of Drought and Heat Stress Tolerance in Wheat (Triticum aestivum L.) Genotypes and Their Response to Selenium Nanoparticles Application

In this study, the role of selenium nanoparticles (SeNPs, 10 mg·L-1) has been investigated in modulating the negative effects of drought and heat stresses on eight bread wheat (Triticum aestivum L.) genotype seedlings. Those genotypes included Giza-168, Giza-171, Misr-1, Misr-3, Shandweel-1, Sids-1, Sids-12, and Sids-14. The study included six treatments as follows: regular irrigation with 100% Field Capacity (FC) at a temperature of 23 ± 3 °C (T1), drought stress with 60% FC (T2), heat stress of 38 °C for 5 h·day-1 (T3), foliar spray of 10 mg·L-1 of SeNPs only (T4), a combination of drought stress with foliar spray of 10 mg·L-1 of SeNPs (T5), and heat stress with foliar spray of 10 mg·L-1 of SeNPs (T6). The experiment continued for 31 days. Foliar application of SeNPs improved the plant growth, morpho-physiological and biochemical responses, and expression of stress-responsive genes in wheat (T. aestivum L.) seedlings. Overall, morpho-physiological traits such as plant height (PH), shoot fresh weight (SFW), shoot dry weight (SDW), root fresh weight (RFW), and root dry weight (RDW) of wheat genotypes grown under different conditions ranged from 25.37-51.51 cm, 3.29-5.15 g, 0.50-1.97 g, 0.72-4.21 g, and 0.11-1.23 g, respectively. From the morpho-physiological perspective, drought stress had a greater detrimental impact on wheat plants than heat stress, whereas heat stress significantly impacted the expression of stress-responsive genes. Stress responses to drought and heat varied between wheat genotypes, suggesting that different genotypes are more resilient to stress. Exogenous spraying of 10 mg·L-1 of SeNPs improved the photosynthetic pigments, photosynthetic rate, gas exchange, and transpiration rate of wheat plants and enhanced drought and heat tolerance by increasing the activity of antioxidant enzymes including catalase (CAT), ascorbate peroxidase (APX), and superoxide dismutase (SOD) and the expression level of stress-responsive genes. Our results showed that spraying wheat seedlings with 10 mg·L-1 of SeNPs enhanced SOD activity for all genotypes as compared to the control, with the Sids-12 genotype having the highest value (196.43 U·mg-1 FW·min-1) and the Giza-168 genotype having the lowest (152.30 U·mg-1 FW·min-1). The expression of PIP1, LEA-1, HSP70, and HSP90 stress-responsive genes was more significant in tolerant genotypes (Giza-171 and Giza-168) than in sensitive ones (Misr-1 and Misr-3) in response to drought and heat stresses. Under stress conditions, the shoot and root fresh weights, photosynthetic pigment content, stomatal conductance (SC), and transpiration rate (TR) were positively correlated with plant height (PH), while root and shoot dry weights, malondialdehyde (MDA), proline, hydrogen peroxide (H2O2), and APX were negatively correlated. Multivariate analysis and biplot results revealed that genotypes Giza-168, Giza-171, Sids-12, and Sids-14 performed well in both stress situations and were classified as stress-tolerant genotypes. These best genotypes may be employed in future breeding projects as tools to face climate change. This study concluded that various physio-biochemicals and gene expression attributes under drought and heat stress could be modulated by foliar application of SeNPs in wheat genotypes, potentially alleviating the adverse effects of drought and heat stress.