Physiological strategies of acute thermal conditions of Rhamdia voulezi collected in the Iguaçu river watershed, Paraná, Brazil: biochemical markers of metabolic and oxidative stress.

Thermal pollution creates substantial challenges that alter energy demand and produce reactive oxygen species that damage fish DNA, proteins, and lipids. Rhamdia voulezi is a species of fish native to the Iguaçu river, Paraná, Brazil, that does not have scientific records of minimum (CTmin) and maximum (CTmax) temperatures required for survival. As it is a top predator species in the food chain and lives at temperatures below 22 °C, the loss of the species can cause functional problems in controlling the ecosystem and energy flow. The study evaluated the tissue metabolism of the brain, heart, and muscle of R. voulezi (n = 72) subjected to acute thermal stress of 31 °C for 2, 6, 12, 24, and 96 h after acclimatization to 21 °C. The biochemical markers SOD, GPx, MDH, HK, and CK of the brain, PCO of the heart and CAT, glycogen, G6PDH, and ALT of muscle were significant. PCA, IBR, thermal sensitive, and condition factor suggested that R. voulezi has different physiological strategies for acclimatization to 31 °C to mobilize and sustain the metabolic needs of oxygenation and energy allocation/utilization for tissue ATP production.

Metabolic responses in the gills of Yellowtail Lambari Astyanax lacustris under low- and high-temperature thermal stress.

OBJECTIVE: Ectothermic fish are directly affected by temperature changes in the environment. The aim of this study was to evaluate the metabolic responses in the gills of Yellowtail Lambari Astyanax lacustris under thermal stress. METHODS: To this end, we used spectrophotometry to evaluate the biomarkers of carbohydrate and protein metabolism, antioxidant defense, and oxidative damage in fish subjected to low (15°C) and high (31°C) temperatures, with control groups held at 23°C, for 2, 6, 12, 24, 48, and 96 h. RESULT: The results showed that cold thermal stress did not change the energy demand, and the antioxidant defense was reduced; therefore, the gills were vulnerable to the action of reactive oxygen species (ROS), presenting increased protein carbonylation at 12 h. With heat thermal stress, a higher energy demand was observed, which was verified by an increase in aerobic metabolism by glycolysis and the citric acid cycle. High-temperature stress also increased the antioxidant defenses, as verified by the increased activities of glutathione peroxidase, glutathione reductase, and glutathione S-transferase. However, the antioxidant defense system could not protect tissues from the action of ROS, as protein carbonylation increased at 6 and 24 h, indicating oxidative stress. CONCLUSION: The results showed that (1) temperature variations caused metabolic adjustments in the gills of Yellowtail Lambari, (2) the adaptive responses were different for winter and summer temperatures, and (3) Yellowtail Lambari recovered homeostasis when subjected to thermal stress, even with the occurrence of oxidative stress.

Assessing physiological responses and oxidative stress effects in Rhamdia voulezi exposed to high temperatures.

Exposure to high temperatures induces changes in fish respiration, resulting in an increased production of reactive oxygen species. This, in turn, affects the enzymatic and non-enzymatic components of antioxidant defenses, which are essential for mitigating cellular stress. Rhamdia voulezi, an economically important fish species endemic to Brazil's Iguaçu River, served as the subject of our study. Our goal was to assess enzymatic antioxidant biomarkers (superoxide dismutase, catalase, glutathione peroxidase, glutathione S-transferase, glutathione reductase, glucose-6-phosphate dehydrogenase), non-protein thiol levels (reduced glutathione), and markers of oxidative damage (lipoperoxidation and carbonylation) in the liver, gills, and kidneys of R. voulezi after acute exposure to high temperatures (31°C) for 2, 6, 12, 24, and 96 h. Control groups were maintained at 21°C. Our findings revealed that the liver exhibited increased superoxide dismutase levels up to 12 h and elevated glutathione S-transferase levels at 12 and 96 h at 31°C. In the gills, superoxide dismutase levels increased up to 24 h, along with increased lipoperoxidation at 2, 6, 12, and 96 h of exposure to high temperatures. The kidneys responded to heat stress at 12 h, with an increase in superoxide dismutase and catalase activity, and lipid peroxidation was observed at 2 and 6 h at 31°C. The three tissues evaluated responded differently to heat stress, with the liver demonstrating greater physiological adjustment to high temperatures. The intricate interplay of various antioxidant defense biomarkers and oxidative damage suggests the presence of oxidative stress in R. voulezi when exposed to high temperatures.

The metabolic response of Araucaria angustifolia embryogenic cells to heat stress is associated with their maturation potential.

Araucaria angustifolia is a critically endangered species and its distribution can be affected by an increase in temperature. In this study, we evaluated the effects of heat stress (30°C) on Araucaria angustifolia cell lines responsive (SE1) and non-responsive (SE6) to the development of somatic embryos. The viability of both cell lines was reduced by heat stress and mitochondria were the organelles most affected. Heat stress for 24h increased the reactive oxygen species (ROS) levels in SE1 cells, followed by a reduction at 48 and 72h. In SE6 cells, an increase occurred after 24 and 48h of stress, returning to control levels at 72h. H2 O2 levels were increased after 24h for both SE1 and SE6 cells, being higher for SE6. Interestingly, at 48 and 72h, H2 O2 levels decreased in SE1 cells, while in SE6, the values returned to the control levels. The respiration of SE6 cells in the presence of oxidisable substrates was inhibited by heat stress, in agreement with the high lipid peroxidation levels. The AaSERK1 gene was identified in both cultures, with greater expression in the SE1 line. Heat stress for 24 and 48h increased gene expression only in this cell line. The activity of peroxidase, superoxide dismutase and enzymes of the glutathione/ascorbate cycle was increased in both cell lines subjected to heat stress. Catalase activity was increased only in SE6 cells at 72h of exposure. These results show that responsive SE1 cells can modulate ROS levels more efficiently than SE6 when these cells are stressed by heat. This ability may be related to the maturation capacity of these cells.

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.

Effects of heat shock on energy metabolism and antioxidant defence in a tropical fish species Psalidodon bifasciatus.

Predictions about global warming have raised interest in assessing whether ectothermic organisms will be able to adapt to these changes. Understanding the physiological mechanisms and metabolic adjustment capacity of fish subjected to heat stress can provide subsidies that may contribute to decision-making in relation to ecosystems and organisms subjected to global climate change. This study investigated the antioxidant defence system and energy metabolism of carbohydrate and protein responses in the gill, liver and kidney tissues of Psalidodon bifasciatus (Garavello & Sampaio 2010), a Brazilian freshwater fish used in aquaculture and in biological studies, following exposure to heat shock at 31°C for 2, 6, 12, 24 and 48 h. The fish presented signs of stress in all tissues tested, as evidenced by increased lipid peroxidation concentration at 2 h and phosphofructokinase, hexokinase and malate dehydrogenase activity at 48 h in the gills; increased glutathione-S-transferase activity at 12 h, citrate synthase activity at 24 h and concentration of reduced glutathione (GSH) concentration at 12 and 48 h in the liver; and through increased activity of superoxide dismutase at 48 h, glutathione reductase at 24 h, glucose-6-phosphate dehydrogenase at 48 h and concentration of GSH at 24 h in the kidney. In the kidneys, changes in the antioxidant system were more prominent, whereas in the gills, there were greater changes in the carbohydrate metabolism. These results indicated the importance of glycolysis and aerobic metabolism in the gills, aerobic metabolism in the liver and pentose-phosphate pathway in the kidneys during homeostasis. The biomarker response was tissue specific, with the greatest number of biomarkers altered in the gills, followed by those in the kidneys and liver.

Gradual increase of temperature trigger metabolic and oxidative responses in plasma and body tissues in the Antarctic fish Notothenia rossii.

Antarctica is considered a thermally stable ecosystem; however, climate studies point to increases in water temperatures in this region. These thermal changes may affect the biological processes and promote metabolic changes in the adapted organisms that live in this region, rendering the animals more vulnerable to oxidative damage. This study assessed the effect of acclimation temperature on the levels of stress response markers in plasma, kidney, gill, liver, and brain tissues of Notothenia rossii subjected to gradual temperature changes of 0.5 °C/day until reaching temperatures of 2, 4, 6, and 8 °C. Under the effect of the 0.5 °C/day acclimation rate, gill tissue showed increased glutathione-S-transferase (GST) activity; kidney tissue showed increased H+-ATPase activity. In the liver, there was also an increase in GSH. In plasma, gradual decreases in the concentrations of total proteins and globulins were observed. These responses indicate a higher production of reactive oxygen species ROS, an imbalance in energy demand, and a lack in protein synthesis. Gradual increase in temperature may cause opposite responses to the thermal shock model in N. rossii.

Cold and warm waters: energy metabolism and antioxidant defenses of the freshwater fish Astyanax lacustris (Characiformes: Characidae) under thermal stress.

Subtropical fish are exposed to seasonal variations in temperature that impose a set of adaptations on their metabolism necessary for the maintenance of homeostasis. In this study, we addressed the effects of temperature variation on the metabolism of Astyanax lacustris, a species of freshwater fish common in the subtropical region of Brazil. Biomarkers of carbohydrate and protein metabolism, antioxidant defense, and oxidative damage were evaluated in the liver of A. lacustris exposed to low (15 °C) and high (31 °C) temperature thermal shock, with controls at 23 °C for 2, 6, 12, 24, 48, 72, and 96 h. A high energy demand was observed during the first 48 h of exposure to 15 °C, which is necessary for metabolic adjustment at low temperatures, with an increase in glycolysis, citric acid cycle, and amino acid catabolism. In addition, at 31 °C, glucose was exported in the first 12 h of exposure, and an increase in the citric acid cycle suggested acetyl-CoA as the pathway substrate, originating from the oxidation of lipids. The antioxidant defenses did not change at 15 °C, as opposed to 31 °C, in which there were changes in several antioxidant defense markers, indicating a response to the production of ROS. However, oxidative stress was observed at both temperatures, with oxidative damage detected by lipid peroxidation at 15 °C and protein carbonylation at 31 °C.

Effect of heat stress on the antioxidant defense system and erythrocyte morphology of Antarctic fishes.

This study analyzed the effect of thermal stress on erythrocytes of Notothenia rossii and Notothenia coriiceps, abundant notothenioids in Admiralty Bay, Antarctic Peninsula. In both species, the antioxidant defense system enzymes, superoxide dismutase, catalase, glutathione peroxidase, glutathione-S transferase, glutathione reductase were punctually altered (8°C for 1, 3 and 6 days) in erythrocytes, indicating that these markers are not ideal for termal stress. However, under the influence of thermal stress, morphological changes in Notothenia coriiceps erythrocytes were observed at all exposure times (1, 3 and 6 days at 8°C), and in Notothenia rossii occurred in 6 days. These results suggest that Notothenia corriceps presents a lower tolerance to thermal stress at 8°C for up to 6 days, since the cellular and nuclear alterations recorded are pathological and may be deleterious to the cells. Among the morphological markers analyzed in this work, we believe that the shape change and nuclear bubble formation may be good stress biomarkers in erythrocytes of Notothenia rossii and Notothenia coriiceps.

Mitochondrial bioenergetics and enzymatic antioxidant defense differ in Paraná pine cell lines with contrasting embryogenic potential.

Araucaria angustifolia is classified as a critically endangered species by the International Union for Conservation of Nature. This threat is worsened by the inefficiency of methods for ex-situ conservation and propagation. In conifers, somatic embryogenesis (SE) associated with cryopreservation is an efficient method to achieve germplasm conservation and mass clonal propagation. However, the efficiency of SE is highly dependent on genotype responsivity to the artificial stimulus used in vitro during cell line proliferation and later during somatic embryo development. In this study, we evaluated the activity of antioxidant enzymes and characterized mitochondrial functions during the proliferation of embryogenic cells of A. angustifolia responsive (SE1) and non-responsive (SE6) to the development of somatic embryos. The activities of the antioxidant enzymes GR (EC 1.6.4.2), MDHAR (EC 1.6.5.4), and POX (EC 1.11.1.7) were increased in SE1 culture, while in SE6 culture, only the activity of DHAR (EC 1.8.5.1) was significantly higher. Additionally, SE6 culture presented a higher number of mitochondria, which agreed with the increased rate of oxygen consumption compared to responsive SE1 culture; however, the mitochondrial volume was lower. Although the ATP levels did not differ, the NAD(P)H levels were higher in SE1 cells. NDs, AOX, and UCP were less active in responsive SE1 than in non-responsive cells. Our results show significant differences between SE1 and SE6 embryogenic cells regarding mitochondrial functions and antioxidant enzyme activities, which may be intrinsic to the in vitro proliferation phase of both cell lines, possessing a crucial role for the induction of in vitro maturation process.

Fish oil supplementation enhances colon recovery after experimental colitis.

INTRODUCTION: Fish oil (FO) has an anti-inflammatory and pro-resolution activity and it has been used to restore physiological disturbances on inflammatory conditions. Here, we investigate whether FO supplementation could, acutely, prevent or restore inflammatory damages on experimental colitis. METHODS: Wistar rats orally received 2 g.kg-1.day-1 of FO for 30 days before induction of experimental colitis. Specimens were collected on the 2nd and 7th days after colitis-induction and intestinal mucus, inflammatory activity and colon integrity were determined. RESULTS: Experimental colitis did cause colon disruption and FO, acutely, did not prevent the loss of intestinal and fecal mucus, neither the increase of inflammatory activity and intestinal permeability. On the 7th day of colitis, FO soften the perturbations of experimental colitis, increasing histological and fecal mucus and, also decreased inflammatory activity, but this was not accompanied by intestinal permeability. CONCLUSION: FO did not protect, acutely, intestinal damages from experimental colitis, but at long run promotes higher intestinal recovery.

Biomarkers of oxidative stress and cell damage in freshwater bivalves Diplodon parodizi exposed to landfill leachate.

Landfill is a public and environmental health problem; establishing and understanding methodologies to decrease its toxicity are thus necessary. Leachate samples were collected, at a sanitary landfill, immediately after the exit from the landfill, i.e. raw leachate (collection point A), after conventional treatment (point B) and after treatment by wetlands (point C). D. parodizi specimens were exposed to 3%, 10% and control (0%) dilutions of leachate from these collection points for 7 days. Markers of antioxidant defences and cell damage were analysed. At point B, the gills of D. parodizi showed higher glutathione-S-transferase (GST) and glutathione reductase (GR) activity; the latter is a supplier of glutathione reductase (GSH). The low GST activity at point A was associated with the hormesis effect. Higher levels of superoxide dismutase (SOD), ethoxyresorufin-O-deethylase (EROD) and glutathione peroxidase (GPx) occurred at point A. Glucose-6-phosphate dehydrogenase (G6PDH) was inhibited at the points with the highest pollutant load and at the highest leachate dilutions. Higher levels of markers at point A may be related to the high pollutant charge and specific compounds present in the untreated leachate. The multi-xenobiotic resistance mechanism (MXR), metallothionein-like proteins (MT) and lipid peroxidation (LPO) did not vary among treatments. The biomarker responses showed negative effects of the leachate on the freshwater bivalve and simultaneously showed that the wetland treatment employed at the Caximba sanitary landfill is effective.

TCTP from Loxosceles Intermedia (Brown Spider) Venom Contributes to the Allergic and Inflammatory Response of Cutaneous Loxoscelism.

LiTCTP is a toxin from the Translationally Controlled Tumor Protein (TCTP) family identified in Loxosceles brown spider venoms. These proteins are known as histamine-releasing factors (HRF). TCTPs participate in allergic and anaphylactic reactions, which suggest their potential role as therapeutic targets. The histaminergic effect of TCTP is related to its pro-inflammatory functions. An initial characterization of LiTCTP in animal models showed that this toxin can increase the microvascular permeability of skin vessels and induce paw edema in a dose-dependent manner. We evaluated the role of LiTCTP in vitro and in vivo in the inflammatory and allergic aspects that undergo the biological responses observed in Loxoscelism, the clinical condition after an accident with Loxosceles spiders. Our results showed LiTCTP recombinant toxin (LiRecTCTP) as an essential synergistic factor for the dermonecrotic toxin actions (LiRecDT1, known as the main toxin in the pathophysiology of Loxoscelism), revealing its contribution to the exacerbated inflammatory response clinically observed in envenomated patients.

Effect of gradual temperature increase on the carbohydrate energy metabolism responses of the Antarctic fish Notothenia rossii.

The warming of the Southern Ocean waters may affect the biological processes and the performance of the fish inhabiting it. The notothenioid group is metabolically specialized to low-temperature environments and may be vulnerable to the climatic changes imposed on the Antarctic continent. However, gradual temperature changes potentially allow an opportunity for plasticity adjustments. The present study evaluated the effect of gradual increase of temperature on the enzymatic and nonenzymatic parameters of energy metabolism in renal, branchial, hepatic, and encephalic tissue of Notothenia rossii subjected to a gradual temperature change of 0.5 °C/day until reaching 2 °C, 4 °C, 6 °C, and 8 °C. Under the effect of an acclimation rate of 0.5 °C/day, the gill tissue showed increased phosphofructokinase (PFK) enzyme activity. In the kidney, there was increased activity of the malate dehydrogenase (MDH), glucose-6-phosphatase (G6PDH), and glycogen phosphorylase (GP) enzymes. There was an increase in lactate concentration in the liver and an increase in GP enzyme activity in the brain. The specific tissue responses indicate the presence of thermal plasticity and an attempt to regulate energy metabolism to mitigate thermal stress in this species under these experimental conditions, possibly through the activation of glycolysis, gluconeogenesis, and glycogenolysis.

Effect of long-term thermal challenge on the Antarctic notothenioid Notothenia rossii.

The thermal stability of the Antarctic Ocean raises questions concerning the metabolic plasticity of Antarctic notothenioids to changes in the environmental temperature. In this study, Notothenia rossii survived 90 days at 8 °C, and their condition factor level was maintained. However, their hepatosomatic (0.29×) index decreased, indicating a decrease in nutrient storage as a result of changes in the energy demands to support survival. At 8 °C, the plasma calcium, magnesium, cholesterol, and triglyceride concentrations decreased, whereas the glucose (1.91×) and albumin (1.26×) concentrations increased. The main energy substrate of the fish changed from lipids to glucose due to a marked increase in lactate dehydrogenase activity, as demonstrated by an increase in anaerobic metabolism. Moreover, malate dehydrogenase activity increased in all tissues, suggesting that fish acclimated at 8 °C exhibit enhanced gluconeogenesis. The aerobic demand increased only in the liver due to an increase (2.23×) in citrate synthase activity. Decreases in the activities of superoxide dismutase, catalase, and glutathione-S-transferase to levels that are most likely sufficient at 8 °C were observed, establishing a new physiological activity range for antioxidant defense. Our findings indicate that N. rossii has some compensatory mechanisms that enabled its long-term survival at 8 °C.

Genome Analysis of Entomopathogenic Bacillus sp. ABP14 Isolated from a Lignocellulosic Compost.

We report the complete genome sequence of Bacillus sp. strain ABP14 isolated from lignocellulosic compost and selected by its ability in hydrolyzing carboxymethyl cellulose. This strain does not produce a Cry-like protein but showed an insecticidal activity against larvae of Anticarsia gemmatalis (Lepidoptera). Genome-based taxonomic analysis revealed that the ABP14 chromosome is genetically close to Bacillus thuringiensis serovar finitimus YBT020. ABP14 also carries one plasmid which showed no similarity with those from YBT020. Genome analysis of ABP14 identified unique genes related to cell surface structures, cell wall, metabolic competence, and virulence factors that may contribute for its survival and environmental adaptation, as well as its entomopathogenic activity.

Cellulose production increases sorghum colonization and the pathogenic potential of Herbaspirillum rubrisubalbicans M1.

Three species of the ß-Proteobacterial genus Herbaspirillum are able to fix nitrogen in endophytic associations with such important agricultural crops as maize, rice, sorghum, sugar-cane and wheat. In addition, Herbaspirillum rubrisubalbicans causes the mottled-stripe disease in susceptible sugar-cane cultivars as well as the red-stripe disease in some sorghum cultivars. The xylem of these cultivars exhibited a massive colonisation of mucus-producing bacteria leading to blocking the vessels. A cluster of eight genes (bcs) are involved in cellulose synthesis in Herbaspirillum rubrisubalbicans. Mutation of bcsZ, that encodes a 1,4-endoglucanase, impaired the exopolysaccharide production, the ability to form early biofilm and colonize sorghum when compared to the wild-type strain M1. This mutation also impaired the ability of Herbaspirillum rubrisubalbicans M1 to cause the red-stripe disease in Sorghum bicolor. We show cellulose synthesis is involved in the biofilm formation and as a consequence significantly modulates bacterial-plant interactions, indicating the importance of cellulose biosynthesis in this process.

Metabolic responses in Antarctic Nototheniidae brains subjected to thermal stress.

Antarctic Nototheniidae is an attractive group for studying metabolic and physiological responses at high temperatures. The present work investigated the metabolic responses of the carbohydrate metabolism and antioxidant system to thermal stress at 8 °C (for 2-144 h) in the brains of Notothenia rossii and Notothenia coriiceps. In N. coriiceps, glycogenolysis was essential in the first hours of exposure (2 h) at 8 °C and, in addition to inhibiting glucose-6-phosphatase activity, was important for activating the pentose phosphate pathway. In N. rossii, anaerobic metabolism was reduced in the first hours of exposure (2 and 6 h) at 8 °C, followed by reduced hexokinase activity, suggesting energy regulation between neurons and astrocytes. The antioxidant system results indicated the importance of the actions of the glutathione-dependent antioxidant enzymes glutathione-S-transferase and glutathione peroxidase as well as those of catalase in N. coriiceps and the action of glutathione-S-transferase, glutathione peroxidase and glutathione reductase in N. rossii, especially during the first 12 h of thermal stress exposure. These results indicate tissue-specific patterns and species-specific responses to this stress.

TCTP from loxosceles intermedia (Brown Spider) venom contributes to the allergic and inflammatory response of cutaneous loxoscelism

LiTCTP is a toxin from the Translationally Controlled Tumor Protein (TCTP) family identified in Loxosceles brown spider venoms. These proteins are known as histamine-releasing factors (HRF). TCTPs participate in allergic and anaphylactic reactions, which suggest their potential role as therapeutic targets. The histaminergic effect of TCTP is related to its pro-inflammatory functions. An initial characterization of LiTCTP in animal models showed that this toxin can increase the microvascular permeability of skin vessels and induce paw edema in a dose-dependent manner. We evaluated the role of LiTCTP in vitro and in vivo in the inflammatory and allergic aspects that undergo the biological responses observed in Loxoscelism, the clinical condition after an accident with Loxosceles spiders. Our results showed LiTCTP recombinant toxin (LiRecTCTP) as an essential synergistic factor for the dermonecrotic toxin actions (LiRecDT1, known as the main toxin in the pathophysiology of Loxoscelism), revealing its contribution to the exacerbated inflammatory response clinically observed in envenomated patients.