Screening and Structural Characterization of Heat Shock Response Elements (HSEs) in Entamoeba histolytica Promoters.

Entamoeba histolytica (E. histolytica) exhibits a remarkable capacity to respond to thermal shock stress through a sophisticated genetic regulation mechanism. This process is carried out via Heat Shock Response Elements (HSEs), which are recognized by Heat Shock Transcription Factors (EhHSTFs), enabling fine and precise control of gene expression. Our study focused on screening for HSEs in the promoters of the E. histolytica genome, specifically analyzing six HSEs, including Ehpgp5, EhrabB1, EhrabB4, EhrabB5, Ehmlbp, and Ehhsp100. We discovered 2578 HSEs, with 1412 in promoters of hypothetical genes and 1166 in coding genes. We observed that a single promoter could contain anywhere from one to five HSEs. Gene ontology analysis revealed the presence of HSEs in essential genes for the amoeba, including cysteine proteinases, ribosomal genes, Myb family DNA-binding proteins, and Rab GTPases, among others. Complementarily, our molecular docking analyses indicate that these HSEs are potentially recognized by EhHSTF5, EhHSTF6, and EhHSTF7 factors in their trimeric conformation. These findings suggest that E. histolytica has the capability to regulate a wide range of critical genes via HSE-EhHSTFs, not only for thermal stress response but also for vital functions of the parasite. This is the first comprehensive study of HSEs in the genome of E. histolytica, significantly contributing to the understanding of its genetic regulation and highlighting the complexity and precision of this mechanism in the parasite's survival.

Integrated biomarker response index as an ally in the observation of metabolic biomarkers in muscle of Astyanax lacustris exposed to thermal variation.

Temperature is an important factor that conditions the physiological responses of fish, generating a stressful condition when in non-ideal parameters. Thus, the objective was to evaluate metabolic aspects in the muscle of fish Astyanax lacustris submitted to thermal shock. The specimens were subjected to 2, 6, 12, 24, 48, and 96 h of exposure to 15 °C and 31 1 °C, with their respective controls 23 °C. At 15 °C there was a reduction in glycogenolysis in the initial periods indicated by changes in glycogen phosphorylase (GP), pyruvate and lactate. Subsequently, there was an increase in GP activity, pyruvate levels and hexokinase activity in the next time periods, suggesting an increase in energy demand. At 31 °C there was observed low need for the protein metabolism, indicated by reduction in the activity of aspartate aminotransferase and alanine aminotransferase. At 15 °C, initial periods show an increase in glutathione reductase activity and an increase in carbonylated protein levels, indicating induction of oxidative stress for muscle. At 31 °C, there was a punctual increase in reduced glutathione levels at 24 h. In addition, the integrated biomarker response index proved to be a good ally in the evaluation of a set of biomarkers, corroborating the results observed by the biomarkers individually. Thus, it is possible to conclude that the acute thermal shock affects the metabolism of A. lacustris muscle, which undergoes rearrangements to deal with temperature, where 15 °C is more stressful than 31 °C.

Can high- and low-temperature thermal stress modulate the antioxidant defense response of Astyanax lacustris brain?

Change in temperature of aquatic environment have impacts on the physiology of fish, especially in the brain, which is a vital organ and prone to oxidative damage. Astyanax lacustris is a freshwater fish that play an important role in the food market and has been increasingly used in fish farms, besides environmental monitoring studies. Therefore, this study aimed to evaluate the responses of antioxidant biomarkers and products of the oxidative process in the brains A. lacustris subjected to thermal shock. The specimens were obtained from artificial farming lakes and subjected to shock induced by exposure to high (31 °C ± 0.5) and low (15 °C ± 0.5) temperature for 2, 6, 12, 24, 48, 72 and 96 h; control group were maintained at 23 °C ± 0.5. At 31 °C, glutathione-related enzymes were more responsive, suggested by the change activity of GPx and G6PDH enzymes, in addition to GSH levels. At 15 °C, enzymes of the first line of defense were more active, evidenced by the change CAT activity. No significant changes were detected in the levels of ROS, LPO and PCO. These results indicate that the brains of A. lacustris have an efficient antioxidant defense system with the ability to acclimatize to the temperatures tested.

Antarctic Lichens under Long-Term Passive Warming: Species-Specific Photochemical Responses to Desiccation and Heat Shock Treatments.

Climate warming in the Antarctic tundra will affect locally dominant cryptogams. Being adapted to low temperatures and freezing, little is known about the response of the polar lichens' primary photochemistry to warming and desiccation. Since 2008, we have monitored the ecophysiological responses of lichens to the future warming scenario during a long-term warming experiment through open top chambers (OTCs) on Fildes Peninsula. We studied the primary photochemical response (potential Fv/Fm and effective efficiency of photosystem II YPSII) of different lichen taxa and morphotypes under desiccation kinetics and heat shock experiments. As lichens grow slowly, to observe changes during warming we methodologically focused on carbon and nitrogen content as well as on the stable isotope ratios. Endemic Himantormia lugubris showed the strongest effect of long-term warming on primary photochemistry, where PSII activity occurred at a lower %RWC inside the OTCs, in addition to higher Fv/Fm values at 30 °C in the heat shock kinetic treatment. In contrast, Usnea aurantiaco-atra did not show any effect of long-term warming but was active at a thallus RWC lower than 10%. Both Cladonia species were most affected by water stress, with Cladonia aff. gracilis showing no significant differences in primary photochemical responses between the warming and the control but a high sensibility to water deficiency, where, at 60% thallus RWC, the photochemical parameters began to decrease. We detected species-specific responses not only to long-term warming, but also to desiccation. On the other hand, the carbon content did not vary significantly among the species or because of the passive warming treatment. Similarly, the nitrogen content showed non-significant variation; however, the C/N ratio was affected, with the strongest C/N decrease in Cladonia borealis. Our results suggest that Antarctic lichens can tolerate warming and high temperature better than desiccation and that climate change may affect these species if it is associated with a decrease in water availability.

Infuence of heat stress on in vitro oocyte and embryo production in high-yielding Holstein cows / Influência do estresse calórico na produção in vitro de oócitos e embriões de vacas Holandesas de alta produtividade

The objective of this study was to evaluate the influence of thermal shock on oocytes used in the production of in vitro embryos (IVP) of high productivity Holstein cows on the day of follicular aspiration (OPU; 0), 30, 60 and 90 days before the OPU. From the mean temperature on day 0 and on the previous 30, 60 and 90 days, they were classifed into comfort group (TC; up to 15°C) and heat stress (HS; above 15°C) groups.Anegative influence was observed on oocytes and viable embryos (total and grade I). The heat stress in the periods of 30 and 60 days prior to OPU resulted in lower production of viable oocytes (P=0.0028; P=0.0092, respectively). Under stress, on the day of OPU (HS-OPU), cows showed no reduction in the amount of viable oocytes (P=0.5497) and there was no influence of temperature for the group stressed 90 days before OPU (P=0.8287). For total embryos, the difference occurred only in the HS-30 group (P=0.0317), where the groups HS-OPU, HS-60, HS-90 presented, respectively, P=0. 1987, P=0.0596 and P=0.4580. Regarding the production of embryos of grade 1, there was no difference for the groups HS-OPU (P=0.2291) and HS-90 (P=0.2868), but there was a reduction for HS-30 (P=0.0143) and HS-60 (P=0.0253). In summary, heat stress had a negative impact when it occurred 30 or 60 days before follicular aspiration. In addition, 30 days seems to be the period of more susceptibility and that causes the greatest deleterious effects on oocyte viability and IVP.(AU)

Objetivou-se avaliar a infuência do estresse térmico em oócitos utilizados na produção in vitro de embriões (PIV) bovinos da raça Holandesa de alta produtividade no dia da aspiração folicular (OPU; 0), 30, 60 e 90 dias antes da OPU. A partir da temperatura média no dia 0 e aos 30, 60 e 90 dias anteriores, foram classificados nos grupos conforto (CT; até 15°C) e estresse por calor (ET -acima de 15°C). Observou-se infuência negativa em oócitos e embriões viáveis (total e grau I). A submissão ao estresse térmico nos períodos de 30 e 60 dias anteriores à OPU resultou em menor produção de oócitos viáveis (P=0,0028; P=0,0092, respectivamente). Sob estresse, no dia da OPU (ET-OPU), as vacas não apresentaram redução na quantidade de oócitos viáveis (P=0,5497) e não houve infuência da temperatura para o grupo estressado 90 dias antes da OPU (P=0,8287). Para embriões totais, a diferença ocorreu apenas no grupo ET-30 (P=0,0317), onde os grupos ET-OPU, ET-60, ET-90 apresentaram, respectivamente, P=0,1987, P=0,0596 e P=0,4580. Em relação à produção de embriões grau 1, não houve diferença para os grupos ET-OPU (P=0,2291) e ET-90 (P=0,2868), porém houve redução para ET-30 (P=0,0143) e ET- 60 (P=0,0253). Em resumo, o estresse por calor teve impacto negativo quando ocorreu 30 ou 60 dias antes da aspiração folicular. Além disso, 30 dias parece ser o período de maior suscetibilidade e que causa os maiores efeitos deletérios na viabilidade oocitária e na PIV.(AU)

Mechanical Properties of Hybrid Carbonized Plant Fibers Reinforced Bio-Based Epoxy Laminates.

In this work, henequen and ixlte plant fibers were carbonized in a horizontal quartz tube furnace. Several carbonized and non-carbonized fiber fabric configurations were impregnated with a bio-based epoxy resin through the infuseon process. The infrared spectra revealed characteristic bands of styrene instead of organic compounds, representing that the carbonization procedure was adequate to carbonize the plant fibers. The porosity volume ratio for the non-carbonized henequen laminates showed the highest number of voids >1.9%, and the rest of the composites had a similar void density between 1.2-1.7%. The storage modulus of the non-carbonized and carbonized henequen laminates resulted in 2268.5 MPa and 2092.1 MPa, respectively. The storage modulus of the carbonized ixtle laminates was 1541.4 MPa, which is 37.8% higher than the non-carbonized ixtle laminates and 12% higher than henequen composites. The laminates were subject to thermal shock cycling, and tomography scans revealed no alterations on the porosity level or in the cracks after the cycling procedure. Thermal shock cycling promoted the post-curing effect by increasing the glass transition temperature. The viscoelastic results showed a variation in the storage modulus when the carbonized fiber fabrics were located between natural fiber fabrics, which was attributed to more excellent compaction during the infusion process. Variations in the viscoelastic behavior were observed between the different types of natural fibers, which influenced the mechanical properties.

Nannochloropsis spp. as Feed Additive for the Pacific White Shrimp: Effect on Midgut Microbiology, Thermal Shock Resistance and Immunology.

This work aimed to evaluate Nannochloropsis spp. as feed additive in the diet of Pacific white shrimp for their effect on midgut microbiology, thermal shock resistance and immunological parameters. Initially, the digestibility of the microalgae meal was assessed, and the apparent digestibility coefficient (ADC) was determined. The ADC was, in general, high in lipids (78.88%) and eicosapentaenoic fatty acid (73.86%). Then, Nannochloropsis spp. were included in diets at four levels (0, 0.5, 1 and 2% inclusion). The shrimp were reared in 500 L clear water tanks containing 20 shrimp per tank with an initial weight of 6.05 ± 0.06 g and fed four times a day. Shrimp fed with supplemented diets containing Nannochloropsis spp. (0.5 and 2%) presented higher resistance to thermal shock when compared to the non-supplemented group (control). Shrimp fed with 1 and 2% of algae inclusion had a higher production of reactive oxygen species (ROS) when compared to other treatments. No statistical difference was observed in the immunological parameters and microbiology of the intestinal tract. Thus, the inclusion of Nannochloropsis spp. in shrimp diets at 0.5 and 2% levels increases resistance to thermal shock and ROS production in shrimp.

Mecanismos moleculares mediadores da citoproteção de células beta pancreáticas induzidos por prolactina / Role of HSPB1 in PRL-induced cytoprotective effects on beta cells

A manutenção da célula de ilhotas in vitro aparece como uma estratégia atraente para aumentar o resultado do transplante de ilhotas pancreáticas. Entretanto, o destino das ilhotas em cultura é determinado pelo equilíbrio entre mediadores pró e antiapoptóticos. Nós mostramos anteriormente que os níveis de HSPB1 são aumentados pela prolactina (PRL) tanto nas células beta pancreáticas humanas quanto nas células de insulinoma murino MIN6. Além disso, mostramos que os efeitos pró- sobrevivência induzidos pela prolactina nas células beta pancreáticas são mediados pela HSPB1. Uma vez que o papel da HSPB1 nas células beta não foi estudado diretamente, procuramos explorar os mecanismos moleculares pelos quais a HSPB1 medeia a citoproteção da célula beta induzida pela PRL. Para isso, células MIN6 derivadas de um insulinoma de camundongo e cultura primária de ilhotas pancreáticas murinas (I), silenciadas ou superexpressando HSPB1 foram submetidas à privação de soro e então pré- tratadas na presença ou na ausência de PRL (300 ng / mL) e expostos a ou citocinas (IL-1ß (0,8 ng / mL), IFN-γ (4 ng / mL) e TNF-α (8 ng / mL) por 16 ou 24 h. Após esses períodos de tempo foi avaliada a viabilidade celular. De fato, as células silenciadas para HSPB1 tiveram maiores porcentagens de morte celular em comparação aos controles. No entanto, a superexpressão de HSPB1 sozinha imita os efeitos citoprotectores da Prolactina em ambas as células MIN6 e nas culturas primárias das ilhotas. Estes resultados mostram o papel fundamental da HSPB1 no efeito citoprotetor inibindo a apoptose inducida pelo tratamento com citocinas pró-inflamatórias. Além disso, os lisados de células Min6 tratadas com citocinas na presença ou na ausência de PRL durante 6 h foram sujeitos a imunoprecipitação de HSPB1. Proteínas coimmunoprecipitadas separadaspor SDS-PAGE e posteriormente identificadas por nano-HPLC acoplado à espectrometria de massas. Células pré-tratadas com PRL apresentaram um enriquecimento de proteínas que coprescipitaram com HSPB1 relacionadas em processos de resistência ao estresse oxidativo, degradação proteica e metabolismo de carboidratos. Células MIN6, silenciadas ou superexpressando HSPB1 foram expostas á menadiona e peróxido de hidrogênio e parâmetros oxidativos foram analisados. O silenciamento de HSPB1 promoveu células mais sensíveis ao estresse oxidativo e levou a uma redução da capacidade antioxidante, enquanto que prolactina induziu citoproteção mediada por HSPB1 contra o estresse oxidativo. A superexpressão de HSPB1, no entanto, levou a efeitos opostos. O tratamento com PRL, o silenciamento ou superexpressão de HSPB1 não mudou a expressão de enzimas antioxidantes, mas os níveis proteicos de HSPB1 estão relacionados com a modulação da razão GSH/GSSG e a atividade de G6PD. Dado de estudos recentes reportam que o perfil respiratório das ilhotas prévias ao transplante pode predizer seu desempenho e que não se sabe nada sobre se a PRL poderia modular a função mitocondrial nas células beta; no presente projeto foi investigado se o tratamento hormonal poderia aumentar a eficiência mitocondrial das células beta. Observamos que o tratamento com citocinas pró-inflamatórias produziu uma diminuição na eficiência do consumo de oxigênio mitocondrial estar relacionado à síntese de ATP. Esses resultados foram significativamente revertidos a valores similares ao obtidos nas células submetidas Às condições de máxima viabilidade após o tratamento com PRL. Além disso, os resultados mostraram que os níveis elevados de HSPB1 medeiam este efeito, uma vez que a falta desta proteína anulou significativamente a recuperação da função mitocondrial induzida pelo tratamento hormonal. Visto que as taxas de síntese de ATP mitocondrial são as responsáveis pela elevação na sua concentração intracelular e que esse evento está diretamente relacionado com a secreção de insulina nas células beta, analisamos se diferentes níveis proteicos de HSPB1 poderia modificar a função secretora de células beta. Para isso foram calculados os índices de estímulo da secreção de insulina em resposta ao aumento da concentraçãode glicose no meio de cultura tanto em células parentais MIN6 como em culturas primárias de ilhotas pancreáticas murinas que foram submetidas ou não ao silenciamento ou superexpressão de HSPB1. Nossos resultados mostraram que nem a presença de citocinas, Prolactina, ou a ausência ou superexpressão de HSPB1 nas culturas celulares analisadas apresentaram diferença significativa em relação aos índices de estímulo da secreção e conteúdo de insulina. Esses resultados sugerem que nem a falta, nem a superexpressão de HSPB1 poderia alterar a função de célula beta. Nós mostramos a relevância da HSPB1 em ambos os efeitos pró- sobrevivência da PRL contra a morte da célula beta induzida tanto por citocinas quanto por indução de estresse oxidativo. Este último efeito poderia também estar relacionado com a participação da HSPB1 na recuperação da função mitocondrial observada após o tratamento hormonal corroborando assim parte dos resultados obtidos nos experimentos de immunoprecipitação. Finalmente, nossos resultados destacam a importância de mais estudos visando um entendimento mais profundo das funções da HSPB1 nas células beta, uma vez que elas poderiam levar à mitigação da morte da célula beta através da regulação positiva de uma via de proteção endógena, que não é dependente da modulação do sistema imunológico

The success of islet transplantation has improved lately. Unfortunately, it is still compromised by cell loss. Maintaining islet cell in vitro appears as an attractive strategy to increase the outcome of pancreatic islet transplantation. However, islet fate in culture is determined by the balance between pro- and anti- apoptotic mediators. We have previously shown that Heat Shock Protein B1 (HSPB1) levels are increased by prolactin (PRL) on both human pancreatic beta cells and MIN6 murine insulinoma cells. Furthermore, we have demonstrated the prolactin-induced pro-survival effects on pancreatic beta-cells are mediated by HSPB1. Since HSPB1 role in beta cells has not been directly studied, we set out to explore the molecular mechanisms by which HSPB1 mediates PRL-induced beta cell cytoprotection. For this purpose, MIN6 insulinoma mouse cells and primary culture of murine pancreatic islets (I) wild type, HSPB1 silenced or overexpressing the chaperone were subjected to serum starvation and then pre-treated in the presence or in the absence of PRL (300 ng/mL) and exposed to or cytokines (IL-1ß (0,8 ng/mL), IFN-γ (4 ng/mL) and TNF-α (8 ng/mL)) for 16 or 24h. Then, we analyse cell viability. HSPB1silenced cells presented higher percentages of cell death compared to controls. However, the overexpression of HSPB1, independently of hormonal treatment, was able mimic the cytoprotective effects of Prolactin. These results point at the key role of HSPB1 in the cytoprotective effect against proinflammatory cytokines-induced beta cell death. In addition, lysates from Min6 cells incubated for 6 hours in the presence of a cocktail of cytokines and/or PRL were subjected to HSPB1 immunoprecipitation. Co-precipitated proteins were identified by SDS-PAGE coupled to mass spectrometry. We found an enrichment of proteins relatedto signaling pathways involved in a response against oxidative and endoplasmic reticulum stress induction. Moreover, we also identified antiapoptotic effects and carbohydrate metabolism related proteins. Indeed, HSPB1 knockdown rendered cells more sensitive to oxidative stress and led to a reduced antioxidant capacity, while prolactin induced an HSPB1- mediated cytoprotection against ROS induced beta-cell apoptosis. One again, HSPB1 overexpression mimic PRL- induced cytoprotection. While hormonal treatment, HSPB1 silencing or overexpression did not change the expression of antioxidant enzymes; this conditions influenced reduced glutathione cell content and G6DP activity. Since recent studies have pointed that islets respiratory profile prior to transplantation may predict their performance; we also investigated whether PRL treatment could increase beta-cell mitochondrial efficiency. We observed a cytokine-induced increase of mitochondrial oxygen consumption rate not related to ATP synthesis, which was significantly decreased upon PRL treatment. HSPB1 was a key mediator of this effect since the lack of this protein significantly abrogated PRL-induced mitochondrial function recovery. The secretory function was then analysed in wild type MIN6 cells as well as in primary cultures of pancreatic islets either HSPB1 silenced or overexpressing the chaperone. Cells were subjected to serum starvation and then pre-treated in the presence or in the absence of PRL and exposed to cytokines for 16 or 24h. We didn´t found significant differences in both glucose induced-insulin secretion and insulin content between the hormonal treatment, HSPB1 silencing or overexpression. These results suggest that neither lack, nor overexpression of HSPB1 could alter beta cell function. Altogether our results have shown the importance of HSPB1 on PRL prosurvival effects as well as on maintenance of mitochondrial efficiency against both cytokine treatment and oxidative-stress-induced beta cell damage. These results are in accordance with the PRL-induced enrichment of HSPB1 interacting proteins displaying functions related to protein degradation, oxidative stress protection or mitochondrial carbohydrate metabolism.Finally, our results outline the importance of further studies aiming at a deeper understanding of HSPB1 functions on beta cells, since they could lead to the mitigation of beta cell death through the up-regulation of an endogenous protective pathway, which is not dependent on the modulation of the immune system

Tetraploidia em Rhamdia quelen (Quoy e Gaimard, 1824) por choque térmico duplo (quente e frio) / Tetraploidy in Rhamdia quelen (Quoy & Gaimard 1824) by a dual thermal shock (heat and cold)

Avaliou-se a eficiência do choque térmico duplo para induzir tetraploidia em jundiá Rhamdia quelen em diferentes tempos pós-fertilização. Vinte mililitros de ovos (10, 15, 20, 25, 30 e 35 mpf minutos pós-fertilização) foram tratados com choque térmico quente (39±0,2ºC) durante 3 min seguido de choque térmico frio (1,0±0,1ºC) durante 30 min. Um grupo controle foi utilizado. As taxas de fertilização e de eclosão foram medidas 12 hpf (horas pós-fertilização) e 30 hpf, respectivamente. Decorridas 60 hpf, 25 larvas de cada unidade experimental foram fixadas para verificação da ploidia, por citometria de fluxo. A taxas de fertilização foram de 87,83% para o controle e de 23,4%; 28,5%; 30,4%; 20,0%; 30,3% e 36,7%, para os grupos tratados com choque térmico aos 10, 15, 20, 25, 30, 35 mpf, respectivamente. Encontraram-se tetraploides apenas nos grupos de ovos tratados aos 15 mpf e 20 mpf. Este é o primeiro trabalho de indução de tetraploidia em jundiá por choques térmicos quente e frio, o qual possibilitou o aperfeiçoamento dessa técnica.(AU)

The efficiency of double thermal shock to induce tetraploidy in Rhamdia quelen (jundiá) at different post-fertilization periods was assessed. Each test was performed with 20 mL of eggs (10, 15, 20, 25, 30 and 35 mpf minutes post-fertilization) exposed to hot thermal shock (39±0.2ºC) for 3 min, followed by cold thermal shock (1.0±0.1ºC) for 30 min. A control group was also employed. Fertilization and hatching rates were evaluated 12 and 30 hours post-fertilization (hpf), respectively. After 60 hpf, twenty-five larvae from each experimental unit were fixed to verify the ploidy by flow cytometry. Control fertilization rate was 87.83% and reached 23.4%, 28.5%, 30.4%, 20.0%, 30.3%, 36.7% respectively for 10, 15, 20, 25, 30, 35 mpf. Tetraploids were detected only at 15 mpf and 20 mpf. This is the first research of tetraploidy in jundiá obtained by hot and cold shocks, making possible the improvement of this technique.(AU)

Tetraploidia em Rhamdia quelen (Quoy e Gaimard, 1824) por choque térmico duplo (quente e frio) / Tetraploidy in Rhamdia quelen (Quoy & Gaimard 1824) by a dual thermal shock (heat and cold)

Avaliou-se a eficiência do choque térmico duplo para induzir tetraploidia em jundiá Rhamdia quelen em diferentes tempos pós-fertilização. Vinte mililitros de ovos (10, 15, 20, 25, 30 e 35 mpf – minutos pós-fertilização) foram tratados com choque térmico quente (39±0,2ºC) durante 3 min seguido de choque térmico frio (1,0±0,1ºC) durante 30 min. Um grupo controle foi utilizado. As taxas de fertilização e de eclosão foram medidas 12 hpf (horas pós-fertilização) e 30 hpf, respectivamente. Decorridas 60 hpf, 25 larvas de cada unidade experimental foram fixadas para verificação da ploidia, por citometria de fluxo. A taxas de fertilização foram de 87,83% para o controle e de 23,4%; 28,5%; 30,4%; 20,0%; 30,3% e 36,7%, para os grupos tratados com choque térmico aos 10, 15, 20, 25, 30, 35 mpf, respectivamente. Encontraram-se tetraploides apenas nos grupos de ovos tratados aos 15 mpf e 20 mpf. Este é o primeiro trabalho de indução de tetraploidia em jundiá por choques térmicos quente e frio, o qual possibilitou o aperfeiçoamento dessa técnica.

The efficiency of double thermal shock to induce tetraploidy in Rhamdia quelen (jundiá) at different post-fertilization periods was assessed. Each test was performed with 20 mL of eggs (10, 15, 20, 25, 30 and 35 mpf – minutes post-fertilization) exposed to hot thermal shock (39±0.2ºC) for 3 min, followed by cold thermal shock (1.0±0.1ºC) for 30 min. A control group was also employed. Fertilization and hatching rates were evaluated 12 and 30 hours post-fertilization (hpf), respectively. After 60 hpf, twenty-five larvae from each experimental unit were fixed to verify the ploidy by flow cytometry. Control fertilization rate was 87.83% and reached 23.4%, 28.5%, 30.4%, 20.0%, 30.3%, 36.7% respectively for 10, 15, 20, 25, 30, 35 mpf. Tetraploids were detected only at 15 mpf and 20 mpf. This is the first research of tetraploidy in jundiá obtained by hot and cold shocks, making possible the improvement of this technique.

Adaptive Responses to Thermal Stress in Mammals / Respuestas de adaptación al estrés térmico en mamíferos / Respostas de adaptação ao estresse térmico em mamíferos

The environment animals have to cope with is a combination of natural factors such as temperature. Extreme changes in these factors can alter homeostasis, which can lead to thermal stress. This stress can be due to either high temperatures or low temperatures. Energy transference for thermoregulation in homoeothermic animals occurs through several mechanisms: conduction, convection, radiation and evaporation. When animals are subjected to thermal stress, physiological mechanisms are activated which may include endocrine, neu-roendocrine and behavioral responses. Activation of the neuroendocrine system affects the secretion ofhormones and neurotransmitters which act collectively as response mechanisms that allow them to adapt to stress. Mechanisms which have developed through evolution to allow animals to adapt to high environmental temperatures and to achieve thermo tolerance include physiological and physical changes in order to reduce food intake and metabolic heat production, to increase surface area of skin to dissipate heat, to increase blood flow to take heat from the body core to the skin and extremities to dissipate the heat, to increase numbers and activity of sweat glands, panting, water intake and color adaptation of integument system to reflect heat. Chronic exposure to thermal stress can cause disease, reduce growth, decrease productive and reproductive performance and, in extreme cases, lead to death. This paper aims to briefly explain the physical and physiological responses of mammals to thermal stress, like a tool for biological environment adaptation, emphasizing knowledge gaps and offering some recommendations to stress control for the animal production system.

El ambiente al que los animales tienen que enfrentarse es una combinación de factores naturales. Cambios extremos en estos factores pueden alterar la homeostasis y conducir al estrés térmico. Este puede resultar tanto de temperaturas altas como de temperaturas bajas. Cuando se somete a los animales a estrés térmico, se activan mecanismos fisiológicos que pueden incluir respuestas endocrinas, neuroendocrinas y de comportamiento. Los mecanismos desarrollados a través de la evolución para que los animales se adapten a temperaturas ambientales altas y alcancen termotolerancia incluyen cambios fisiológicos y físicos, a fin de reducir la ingesta de alimentos y la producción del calor metabólico; de aumentar el área superficial de la piel para disipar el calor; de aumentar el flujo de sangre para llevar el calor del núcleo del cuerpo a la piel y las extremidades para disipar el calor, o de aumentar el número y la actividad de las glándulas sudoríparas, el jadeo, la ingesta de agua y la adaptación del color del sistema de tegumento para reflejar el calor. La exposición crónica al estrés térmico puede causar enfermedad, reducir el crecimiento, disminuir el rendimiento productivo y reproductivo y, en casos extremos, llevar a la muerte. El objetivo de este trabajo es explicar las respuestas físicas y fisiológicas de los mamíferos al estrés térmico, como una herramienta para la adaptación al entorno biológico, haciendo hincapié en las lagunas de nuestro conocimiento y ofreciendo algunas recomendaciones para controlar el estrés en el sistema de producción animal.

O ambiente ao que os animais têm que enfrentar-se é uma combinação de fatores naturais. Mudanças extremas nestes fatores podem alterar a homeostase e levar ao estresse térmico. Este pode resultar tanto de temperaturas altas quanto de temperaturas baixas. Quando se submete os animais a estresse térmico, se ativam mecanismos fisiológicos que podem incluir respostas endócrinas, neuroendócrinas e de comportamento. Os mecanismos desenvolvidos através da evolução para que os animais se adaptem a temperaturas ambientais altas e alcancem termo tolerância incluem mudanças fisiológicas e físicas, visando reduzir a ingesta de alimentos e a produção do calor metabólico; de aumentar a área superficial da pele para dissipar o calor; de aumentar o fluxo de sangue para levar o calor do núcleo do corpo à pele e às extremidades para dissipar o calor, ou de aumentar o número e a atividade das glândulas sudoríparas, o arquejo, a ingestão de água e a adaptação da cor do sistema de tegumento para refletir o calor. A exposição crônica ao estresse térmico pode causar doença, reduzir o crescimento, diminuir o rendimento produtivo e reprodutivo e, em casos extremos, levar à morte. O objetivo deste trabalho é explicar as respostas físicas e fisiológicas dos mamíferos ao estresse térmico, como uma ferramenta para a adaptação ao entorno biológico, enfatizando nas lagoas de nosso conhecimento e oferecendo algumas recomendações para controlar o estresse no sistema de produção animal.

Motilidade espermática no sêmen suíno adicionado de gema de ovo de quatro linhagens de galinhas / Sperm motility in the swine semen added of egg yolk of four line of chickens

Objetivou-se testar a gema de ovo de quatro diferentes linhagens de galinhas, adicionada ao diluente Beltsville Thawing Solution (BTS), para verificar sua proteção sobre a célula espermática contra o choque térmico. Foram utilizados ejaculados de quatro reprodutores suínos com idade entre 12 e 24 meses, durante 8 semanas (n=32), em um delineamento experimental de blocos ao acaso e esquema fatorial 5x2 (cinco diluentes e duas temperaturas). Ao BTS, adicionou-se 2,5% de gema de ovo de galinhas das linhagens: Plymounth Rock Barrada (PRB); Postura Comercial (PC); Gigante Negra do Pescoço Pelado (GnPP) e Hubbart (Hubb). Os ejaculados foram submetidos aos seguintes tratamentos: 01. A (BTS-controle); 02. B (BTS+PRB); 03. C (BTS+PC); 04. D (BTS+GnPP); 05. E (BTS+Hubb) e avaliados, quanto ao vigor e motilidade, durante cinco dias. Valores do vigor espermático a 17 °C foram semelhantes entre os tratamentos até o D1. A partir do D2, melhores valores de vigor foram observados nos diluentes A e B. Já no sêmen a 4 °C, não foram verificadas diferenças, em relação ao vigor, entre os tratamentos durante o período avaliado. A partir do D0 observou-se queda nos valores de vigor, em todos os tratamentos. Na motilidade espermática a 17 °C, somente a partir do D2, o diluente A manteve maiores valores. No sêmen a 4 °C, não foram observadas diferenças significativas entre os diluentes testados, quanto à motilidade espermática. Concluiu-se que a gema de ovo na concentração de 2,5 % não conferiu proteção às células espermáticas contra o choque térmico, sendo necessários maiores estudos para uma definição da concentração ideal, que permita a manutenção da viabilidade espermática.

The objective of this study was to test the egg yolk of four different lines of chickens added to the Beltsville Thawing Solution and to verify its protection against the sperm cell thermal shock. Ejaculates of four boars aged between 12 and 24 months, during 8 weeks (n=32) in a randomized block design, in a 5x2 factorial scheme (five extenders and two temperatures). To the BTS extender was added 2.5% egg yolk from lines of chickens: Plymounth Rock Barrada (PRB); Business Posture (PC); Black Giant of the Peeled Neck (GnPP) and Hubbart (Hubb). The ejaculates were submitted to the following treatments: 01. A (BTS-control); 02. B (BTS+PRB); 03. C (BTS+PC); 04. D (BTS+GnPP); 05. E (BTS+Hubb) and evaluated for vigor and sperm motility for five days. Sperm vigor values at 17 °C were similar between treatments up to D1, since D2, the best vigour values were observed in the extenders A and B. In the case of semen stored at 4 °C, no differences were observed in relation to the vigor between the treatments during the evaluated period. From the D0, vigour values were observed in all treatments. In relation to the sperm motility at 17 °C, only from D2, the diluent A maintained higher values in relation to the other diluents. In the semen at 4 °C, no significant differences were observed between the extenders tested for sperm motility. It was concluded that 2.5% egg yolk did not protect the sperm cells against thermal shock, and further studies are needed to define the optimal concentration to maintain sperm viability.

Motilidade espermática no sêmen suíno adicionado de gema de ovo de quatro linhagens de galinhas / Sperm motility in the swine semen added of egg yolk of four line of chickens

Objetivou-se testar a gema de ovo de quatro diferentes linhagens de galinhas, adicionada ao diluente Beltsville Thawing Solution (BTS), para verificar sua proteção sobre a célula espermática contra o choque térmico. Foram utilizados ejaculados de quatro reprodutores suínos com idade entre 12 e 24 meses, durante 8 semanas (n=32), em um delineamento experimental de blocos ao acaso e esquema fatorial 5x2 (cinco diluentes e duas temperaturas). Ao BTS, adicionou-se 2,5% de gema de ovo de galinhas das linhagens: Plymounth Rock Barrada (PRB); Postura Comercial (PC); Gigante Negra do Pescoço Pelado (GnPP) e Hubbart (Hubb). Os ejaculados foram submetidos aos seguintes tratamentos: 01. A (BTS-controle); 02. B (BTS+PRB); 03. C (BTS+PC); 04. D (BTS+GnPP); 05. E (BTS+Hubb) e avaliados, quanto ao vigor e motilidade, durante cinco dias. Valores do vigor espermático a 17 °C foram semelhantes entre os tratamentos até o D1. A partir do D2, melhores valores de vigor foram observados nos diluentes A e B. Já no sêmen a 4 °C, não foram verificadas diferenças, em relação ao vigor, entre os tratamentos durante o período avaliado. A partir do D0 observou-se queda nos valores de vigor, em todos os tratamentos. Na motilidade espermática a 17 °C, somente a partir do D2, o diluente A manteve maiores valores. No sêmen a 4 °C, não foram observadas diferenças significativas entre os diluentes testados, quanto à motilidade espermática. Concluiu-se que a gema de ovo na concentração de 2,5 % não conferiu proteção às células espermáticas contra o choque térmico, sendo necessários maiores estudos para uma definição da concentração ideal, que permita a manutenção da viabilidade espermática.(AU)

The objective of this study was to test the egg yolk of four different lines of chickens added to the Beltsville Thawing Solution and to verify its protection against the sperm cell thermal shock. Ejaculates of four boars aged between 12 and 24 months, during 8 weeks (n=32) in a randomized block design, in a 5x2 factorial scheme (five extenders and two temperatures). To the BTS extender was added 2.5% egg yolk from lines of chickens: Plymounth Rock Barrada (PRB); Business Posture (PC); Black Giant of the Peeled Neck (GnPP) and Hubbart (Hubb). The ejaculates were submitted to the following treatments: 01. A (BTS-control); 02. B (BTS+PRB); 03. C (BTS+PC); 04. D (BTS+GnPP); 05. E (BTS+Hubb) and evaluated for vigor and sperm motility for five days. Sperm vigor values at 17 °C were similar between treatments up to D1, since D2, the best vigour values were observed in the extenders A and B. In the case of semen stored at 4 °C, no differences were observed in relation to the vigor between the treatments during the evaluated period. From the D0, vigour values were observed in all treatments. In relation to the sperm motility at 17 °C, only from D2, the diluent A maintained higher values in relation to the other diluents. In the semen at 4 °C, no significant differences were observed between the extenders tested for sperm motility. It was concluded that 2.5% egg yolk did not protect the sperm cells against thermal shock, and further studies are needed to define the optimal concentration to maintain sperm viability.(AU)

Intermação: uma síndrome emergencial: revisão de literatura / Heat Stroke: a emergency syndrome: a review

Os distúrbios relacionados à homeostasia térmica são freqüentes na clínica de pequenos animais, sendo sua etiologia principalmente relacionada a processos infecciosos e inflamatórios. No entanto, há uma forma de hipertermia conhecida por intermação, onde o ponto referencial de temperatura no hipotálamo encontra-se normal e a termorregulação é perdida, devido à deficiência dos mecanismos responsáveis pela dissipação de calor. O excesso de calor produzido é responsável por elevações da temperatura corporal central que podem atingir 43ºC ou mais, resultando em lesão celular, morte celular, disfunções orgânicas e morte do animal, requerendo, dessa forma, intervenção emergencial

Disorders related to thermal homeostasis are common in the small animal clinical, whose etiology is mainly related to infectious and inflammatory processes. However, there is a form of hyperthermia known Heat stroke where the reference point of the hypothalamic temperature is normal and thermoregulation is lost due to deficiency of the mechanisms responsible for heat dissipation. Excess heat is responsible for elevations in core body temperature that can reach 43 degrees or more, resulting in cell damage, cell death, organ dysfunction and death of the animal, requiring thus emergency intervention

Intermação: uma síndrome emergencial: revisão de literatura / Heat Stroke: a emergency syndrome: a review

Os distúrbios relacionados à homeostasia térmica são freqüentes na clínica de pequenos animais, sendo sua etiologia principalmente relacionada a processos infecciosos e inflamatórios. No entanto, há uma forma de hipertermia conhecida por intermação, onde o ponto referencial de temperatura no hipotálamo encontra-se normal e a termorregulação é perdida, devido à deficiência dos mecanismos responsáveis pela dissipação de calor. O excesso de calor produzido é responsável por elevações da temperatura corporal central que podem atingir 43ºC ou mais, resultando em lesão celular, morte celular, disfunções orgânicas e morte do animal, requerendo, dessa forma, intervenção emergencial(AU)

Disorders related to thermal homeostasis are common in the small animal clinical, whose etiology is mainly related to infectious and inflammatory processes. However, there is a form of hyperthermia known Heat stroke where the reference point of the hypothalamic temperature is normal and thermoregulation is lost due to deficiency of the mechanisms responsible for heat dissipation. Excess heat is responsible for elevations in core body temperature that can reach 43 degrees or more, resulting in cell damage, cell death, organ dysfunction and death of the animal, requiring thus emergency intervention(AU)