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The recognition images of cattle brand in an automatic way is a necessity to governmental organs responsible for this activity. To help this process, this work presents a method that consists in using Bag of Visual Words for extracting of characteristics from images of cattle brand and Support Vector Machines Multi-Class for classification. This method consists of six stages: a) select database of images; b) extract points of interest (SURF); c) create vocabulary (K-means); d) create vector of image characteristics (visual words); e) train and sort images (SVM); f) evaluate the classification results. The accuracy of the method was tested on database of municipal city hall, where it achieved satisfactory results, reporting 86.02% of accuracy and 56.705 seconds of processing time, respectively.

Chlorothalonil is an active biocide applied in antifouling paints, and also used as fungicide in agricultural activities with the purpose to protect plants from foliar and seed diseases. Thus, the aim of this study was to evaluate the effects of chlorothalonil exposure on biochemical biomarkers of oxidative metabolism as well as on cholinesterases in the estuarine polychaete Laeonereis acuta. Animals were exposed for 24 and 96 h to the following nominal concentrations of chlorothalonil: 0.1, 10.0 and 100.0 μg/L. The antioxidant capacity against peroxyl radicals (ACAP) and the activity of the enzymes catalase (CAT), superoxide dismutase (SOD), glutathione S-transferase (GST), glutamate cysteine ligase (GCL), acetylcholinesterase (AChE) and propionylcholinesterase (PChE) were evaluated in whole-body tissue. In addition, the levels of reduced glutathione (GSH), lipid peroxidation (LPO), glycogen and lactate levels were also analyzed. A reduction in ACAP levels was observed in animals exposed to the higher chlorothalonil concentration, concomitantly with an induction of GST activity as well as diminution in GSH content in these animals. This disturbance in the redox state of animal tissues leads to an oxidative stress situation, resulting in an induction in LPO levels. It was also demonstrated that chlorothalonil exposure causes alteration in AChE activity, possibly related to damage to membrane lipids. These results demonstrated that chlorothalonil possesses harmful effects to estuarine animals and its use as antifouling biocide has to be carefully reconsidered in risk analysis studies.

Reproductive effects caused by the exposure to environmentally relevant dosages of atrazine on wild animals are poorly understood. This study evaluated the effects of three dosages of atrazine on sperm parameters of adult Calomys laucha males. Adult mice were orally exposed to dosages of 0 (water and vehicle control), 0.1, 1, and 10 mg/kg of animal weight for a 21-day period. Following exposure, analyses were performed to determine sperm motility parameters, plasma membrane integrity and fluidity, mitochondrial functionality, acrosome integrity, DNA damage, lipid peroxidation, and production of reactive oxygen species (ROS) in the sperm samples. Total and progressive motility were reduced in all dosages in comparison to control groups. Membrane integrity and mitochondrial functionality of sperm were reduced in all dosages, and the sperm membrane fluidity increased in the higher dosages of atrazine (1 and 10 mg/kg), in comparison with the vehicle control. A decrease in the acrosome integrity was noted at 10 mg/kg of atrazine, compared to the control groups. The integrity of DNA, ROS generation, and lipid peroxidation of sperm showed no significant differences when compared with the control groups. These results suggest that exposure to low dosages of atrazine can affect sperm parameters of Calomys laucha and therefore reduce the reproductive capacity of wild rodent species.

It has been demonstrated that glyphosate-based herbicides are toxic to animals. In the present study, reactive oxygen species (ROS) generation, antioxidant capacity against peroxyl radicals (ACAP), and lipid peroxidation (LPO), as well as the activity and expression of the acetylcholinesterase (AChE) enzyme, were evaluated in Danio rerio males exposed to 5 or 10 mg/L of glyphosate for 24 and 96 h. An increase in ACAP in gills after 24 h was observed in the animals exposed to 5 mg/L of glyphosate. A decrease in LPO was observed in brain tissue of animals exposed to 10 mg/L after 24 h, while an increase was observed in muscle after 96 h. No significant alterations were observed in ROS generation. AChE activity was not altered in muscles or brains of animals exposed to either glyphosate concentration for 24 or 96 h. However, gene expression of this enzyme in the brain was reduced after 24 h and was enhanced in both brain and muscle tissues after 96 h. Thus, contrary to previous findings that had attributed the imbalance in the oxidative state of animals exposed to glyphosate-based herbicides to surfactants and other inert compounds, the present study demonstrated that glyphosate per se promotes this same effect in zebrafish males. Although glyphosate concentrations did not alter AChE activity, this study demonstrated for the first time that this molecule affects ache expression in male zebrafish D. rerio.

Several studies suggest that Antarctic endemic stenothermic fish species, such as Notothenia coriiceps, have a limited ability to acclimate to warmer conditions. In the present study, we evaluated the effects of exposure to short- and long-term temperature increases on the occurrence of erythrocytic nuclear abnormalities (ENAs) and changes in the leukocyte profile of N. coriiceps. For the experiments, 30 specimens of N. coriiceps were captured in Admiralty Bay and transferred to the Commandant Ferraz Brazilian Antarctic Station where they were acclimated to water at 0 °C for 6 days and subsequently subjected to treatments of temperature increases (2–4 °C) on a short- (1 day) or long- (6 days) term basis. At the end of exposure, peripheral blood samples were collected and blood smears were prepared for qualitative and quantitative analysis of the ENAs and the leukocyte profile. Significant responses were observed for both ENAs and leukocyte profile. The responses of ENAs were characterized by an increased incidence of bilobed nuclei in erythroblasts following 1-day exposure to water at 2–4 °C, as well as increased incidence of bilobed nuclei in erythrocytes following 1-day and 6-day exposure to water at 4 °C. In the leukocyte profile, 1-day exposure to water at 2 °C promoted a stress response characterized by diminished levels of circulating lymphocytes (lymphopenia) and elevated levels of circulating monocytes (monocytosis). In conclusion, our results indicate that elevation of water temperature during short periods provoked cellular and physiological stress in N. coriiceps. Furthermore, we conclude that in terms of nuclear alterations, erythrocytes and erythroblasts respond differently to thermal stress. Finally, we conclude that the methodology proposed was efficient in quantifying thermal stress in N. coriiceps and we recommend the utilization of this technique in the biomonitoring of thermal stress in this species and other notothenioids.

In biofloc technology (BFT) systems, nutrient cycling for microorganisms takes place. This results in minimal or null water exchange. The sum of fish biomass and microorganisms favors alkalinity consumption and, consequently, pH reduction. It is in this context that the present study evaluates alkalinity and pH maintenance using sodium bicarbonate (NaHCO3), calcium carbonate (CaCO3) or calcium hydroxide (Ca(OH)2) on BFT during a Nile tilapia (Oreochromis niloticus) nursery. In this study, 25 fishes/tank was distributed in nine experimental units (useful vol. 37.5 L) and the performance was evaluated at 60 days. During the assay, the temperature, oxygen and pH were maintained within the ideal range levels for Nile tilapia growth. All alkalizing compounds were able to pH and alkalinity correction, but when using NaHCO3, the alkalinity and pH were more elevated than the other treatments. Furthermore, at the beginning these assay, the total ammonia (TAN; NH3 + NH4+) and NO2−-N accumulate and it caused a peak, but mostly experiment remained to very low levels because of the total nitrification activity, resulting in NO3−-N accumulation. Because the non water exchange, at the final experiment the ion Na+ accumulate when utilized NaHCO3, resulting in level similar to brackish water. While using CaCO3 or Ca(OH)2, the Ca2 + ion accumulate, resulting in extremely hard water. Despite this, the fish survival was similar between treatments (about 80%). Moreover, the final weight, daily growth rate and net yield for NaHCO3 and Ca(OH)2 they were higher than CaCO3 treatment. This may have been because of the higher total suspended solids (TSS) and lower protein content of the bioflocs in this treatment. In order to assess the possible physiologic alterations of the fish associated with the production system, the hematocrit, glycemia and plasmatic osmolality were evaluated. Furthermore, the antioxidant capacity against peroxy radicals (ACAP), lipid peroxidation (LPO) and catalase (CAT) and superoxide dismutase (SOD) activities on the gills and liver were also evaluated. There were no differences in biochemical/physiological parameters when the different alkalizing compounds were utilized. The results demonstrate that the use of sodium bicarbonate, hydroxide or calcium carbonate is effective on the alkalinity and pH adjustments of the final proportion of 14.64 ± 0.49, 7.18 ± 0.32 e 24.09 ± 2.32% in relation to the feed consumption, respectively. Thus, the study demonstrates that the use of NaHCO3 and Ca(OH)2 are recommended for alkalinity and pH correction during Nile tilapia nursery on BFT systems, because of the higher growth and net yield, and this sum to less amount of these compounds may represent important economic gain. Statement of relevance The results of this manuscript demonstrate: − Amount of alkalizing compound utilized for maintenance of alkalinity and pH − Best growth to utilize NaHCO3 and Ca(OH)2 − The importance of evaluate the ionic concentration, specially Na+ and Ca+ 2, because the imbalance ionic − The difficulty to solids control in BFT system.

The objective of the present study was to determine the effects of Roundup on oxidative status in adult Danio rerio liver and gills. Reactive oxygen species (ROS) and antioxidant capacity (ACAP) were measured in fish after exposure to Roundup (5 and 10 mg/L) for 24, 48, 72, and 96 h. Furthermore, gene expression related to antioxidant response was evaluated after 24 and 96 h. In gills, an increase in ACAP was observed after 96 h in the highest concentration. In the liver, a reduction in ROS and ACAP was observed after 24 h, whereas an increase in ACAP was observed after 48 h in the highest concentration. Exposure to the lowest concentration caused a reduction in ROS after 72 and 96 h. Regarding gene expression, a reduction in superoxide dismutase 2 (sod2) and glutathione S-transferase (gstπ) was observed. An increase in uncoupling protein 1 (ucp1) expression was observed in gills of animals exposed to the highest concentration after 24 h. Glutathione peroxidase (gpx) gene expression was reduced in the gills of animals exposed to the lowest concentration; however, it was induced in liver tissue after 96 h of exposure to the highest concentration. These results indicate that zebrafish exposure to Roundup alters oxidative status and causes a response in terms of antioxidant defense system gene expression.

In this study, a sensitive HPLC-UV assay was developed and validated for the determination of LASSBio-1736 in rat plasma with sodium diclofenac as internal standard (IS). Liquid-liquid extraction using acetonitrile was employed to extract LASSBio-1736 and IS from 100 μL of plasma previously basified with NaOH 0.1 M. Chromatographic separation was carried on Waters Spherisorb(®) S5 ODS2 C18 column (150 × 4.6 mm, 5 μm) using an isocratic mobile phase composed by water with triethylamine 0.3% (pH 4), methanol and acetonitrile grade (45:15:40, v/v/v) at a flow rate of 1 mL/min. Both LASSBio-1736 and IS were eluted at 4.2 and 5 min, respectively, with a total run time of 8 min only. The lower limit of quantification was 0.2 μg/mL and linearity between 0.2 and 4 μg/mL was obtained, with an R(2) > 0.99. The accuracy of the method was >90.5%. The relative standard deviations intra and interday were

Atrazine is an extensively used herbicide, and has become a common environmental contaminant. Effects on dopaminergic neurotransmission in mammals following exposure to atrazine have been previously demonstrated. Here, the effects of atrazine regarding behavioural and dopaminergic neurotransmission parameters were assessed in the fruit fly D. melanogaster, exposed during embryonic and larval development. Embryos (newly fertilized eggs) were exposed to two atrazine concentrations (10μM and 100μM) in the diet until the adult fly emerged. Negative geotaxis assay, as well as exploratory behaviour, immobility time and number of grooming episodes in an open field system were assessed. Tyrosine hydroxylase (TH) activity and gene expression of the dopaminergic system were also evaluated in newly emerged male and female flies. All analyzed parameters in male flies were not significantly affected by atrazine exposure. However female flies exposed to atrazine at 10μM presented an increase in immobility time and a reduction in exploratory activity in the open field test, which was offset by an increase in the number of grooming episodes. Also, female flies exposed to 100μM of atrazine presented an increase in immobility time. Gene expression of DOPA decarboxylase and dopamine (DA) receptors were also increased only in females. The behavioural effects of atrazine exposure observed in female flies were due to a disturbance in the dopaminergic system.

The herbicide atrazine has been used worldwide with subsequent residual contamination of water and food, which may cause adverse effects on non-target organisms. Animal exposure to this herbicide may affect development, reproduction and energy metabolism. Here, the effects of atrazine regarding survival and redox metabolism were assessed in the fruit fly D. melanogaster exposed during embryonic and larval development. The embryos (newly fertilized eggs) were exposed to different atrazine concentrations (10μM and 100μM) in the diet until the adult fly emerged. Pupation and emergence rates, developmental time and sex ratio were determined as well as oxidative stress parameters and gene expression of the antioxidant defence system were evaluated in newly emerged male and female flies. Atrazine exposure reduced pupation and emergence rates in fruit flies without alterations to developmental time and sex ratio. Different redox imbalance patterns were observed between males and females exposed to atrazine. Atrazine caused an increase in oxidative damage, reactive oxygen species generation and antioxidant capacity and decreased thiol-containing molecules. Further, atrazine exposure altered the mRNA expression of antioxidant genes (keap1, sod, sod2, cat, irc, gss, gclm, gclc, trxt, trxr-1 and trxr-2). Reductions in fruit fly larval and pupal viability observed here are likely consequences of the oxidative stress induced by atrazine exposure.

Antioxidant defense system (ADS) and oxidative stress parameters were evaluated in the Antarctic fish Notothenia rossii and N. coriiceps exposed to increasing temperature. Acclimated fish were kept at 0 °C or exposed to 4 °C for 1 day (N. rossii) or to 2 and 4 °C for 1 and 6 days (N. coriiceps). Measurements were assessed in brain, gills, liver, white muscle and erythrocytes. Parameters analyzed included antioxidant capacity against peroxyl radicals (ACAP); reduced glutathione (GSH) and metallothionein-like proteins (MTLP) concentration; superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione-S-transferase (GST) and glutamate-cysteine ligase (GCL) activity; lipid peroxidation (LPO) level and protein carbonyl (PC) concentration. Increased liver GST activity was observed in N. rossii exposed to 4 °C for 1 day. Increased muscle GPx activity was observed in N. coriiceps after exposure to 2 °C for 1 day. Reduced gill GPx activity and increased liver SOD activity were observed after exposure to 4 °C for 1 day. In N. coriiceps, increased gill GCL activity and reduced gill GPx activity, as well as reduced liver MTLP were observed after exposure to 2 °C for 6 days. Reduced brain SOD activity and increased brain LPO; reduced gill ACAP, GSH concentration and GPx activity, as well as increased gill GCL activity; reduced liver ACAP, MTLP, SOD activity, GST activity and increased liver and erythrocytes LPO were observed after exposure to 4 °C for 6 days. These findings indicate that ADS is more responsive to short-term increasing temperature in the sluggish N. coriiceps than in the active N. rossii. However, responses of N. coriiceps to long-term increasing temperature were transient and did not prevent tissue oxidative damage. Considering the predicted increase in temperature in the Southern Ocean over the next decades, our findings suggest that Antarctic fishes are sensitive to ocean warming, displaying tissue oxidative damage associated with the thermal stress.

Adaptive responses of antioxidant defense systems (ADS) to changes in increased levels of activity are critical, especially in Antarctic fishes. The benthopelagic marbled notothen (Notothenia rossii) shows higher spontaneous activity than the benthonic and sluggish rockcod (N. coriiceps). Therefore, we hypothesize that species-related responses of ADS would occur to counteract different rates of reactive oxygen species formation in these two Antarctic fish. Here we evaluated ADS and oxidative damage in tissues (brain, gills, liver and white muscle) of the two Antarctic fish. Despite no significant differences in lipid and protein oxidative damage were observed, we actually found species- and tissue-specific differences in ADS. Gill metallothionein-like proteins (MTLP) and liver reduced glutathione (GSH) concentrations were higher in N. coriiceps than in N. rossii. Brain and gill antioxidant capacity against peroxyl radicals (ACAP); gill enzyme [glutamate-cysteine ligase (GSL), superoxide dismutase (SOD) and catalase (CAT)] activity; liver GCL and SOD activity; and white muscle CAT activity were higher in N. rossii than in N. coriiceps. Therefore, the more active fish (N. rossii) maintains higher activities of enzymes involved in superoxide ions (O2.-) detoxification and GSH production in peripheral tissues (gills, liver and white muscle). This allows the more active fish (N. rossii) to keep levels of lipid and protein oxidative damage similar to those observed in the sluggish fish (N. coriiceps). It is worth noting that the more active fish also shows a higher brain antioxidant capacity, which could involve other non-enzymatic antioxidants like vitamins C and E. In contrast, N. coriiceps shows lower consumption of non-enzymatic antioxidants in peripheral tissues than N. coriiceps. As hypothesized, our results indicate that differences in ADS profiles between fish species are likely related to their habits and metabolic rates. This would imply in different fish abilities to deal with oxidative stress associated with increasing seawater temperature.

Glyphosate based herbicides, including Roundup, are widely employed in agriculture and urban spaces. The objective of this study was to evaluate the toxicological effects of Roundup on the estuarine polychaeta Laeonereis acuta. Biomarkers of oxidative stress as well as acetylcholinesterase and propionilcholinesterase activities were analyzed. Firstly, the LC50 96h for L. acuta was established (8.19mg/L). After, the animals were exposed to two Roundup concentrations: 3.25mg/L (non-observed effect concentration - NOEC) and 5.35mg/L (LC10) for 24h and 96h. Oxygen consumption was determined and the animals were divided into three body regions (anterior, middle and posterior) for biochemical analysis. An inhibition of both cholinesterase isoforms were observed in animals exposed to both Roundup concentrations after 96h. A significant reactive oxygen species (ROS) reduction was observed in the posterior region of animals in both periods, while antioxidant capacity against peroxyl radicals (ACAP) was reduced in the posterior region of animals exposed for 24h. Considering the antioxidant defense system, both GSH levels and enzyme activities (catalase, superoxide dismutase, glutathione s-transferase, glutathione peroxidase and glutamate cysteine ligase) were not altered after exposure. Lipid peroxidation was reduced in all analyzed body regions in both Roundup concentrations after 24h. Animals exposed to the highest concentration presented a reduction in lipid peroxidation in the anterior region after 96h, while animals exposed to the lowest concentration presented a reduction in the middle region. Overall results indicate that Roundup exposure presents toxicity to L. acuta, causing a disruption in ROS and ACAP levels as well as affects the cholinergic system of this invertebrate species.

Glyphosate is a non-selective and post-emergent herbicide that affects plant growth. Animal exposure to this herbicide can lead to adverse effects, such as endocrine disruption, oxidative stress and behavioural disorders. Drosophilids have been utilized previously as an effective tool in toxicological tests. In the present study, the effects of a glyphosate-based herbicide (Roundup [Original]) were investigated regarding oxidative stress, the antioxidant defence system and acetylcholinesterase (AChE) activity in Drosophila melanogaster. Flies (of both genders) that were 1 to 3days old were exposed to different glyphosate concentrations (0.0g/L=control, 1.0g/L, 2.0g/L, 5.0g/L and 10.0g/L) [corrected] in the diet for 24h and 96h. After the exposure periods, reactive oxygen species (ROS) levels, antioxidant capacity against peroxyl radicals (ACAP) and lipid peroxidation (LPO) levels were quantified. In addition, the mRNA expression of antioxidant genes (i.e., keap1, sod, sod2, cat, irc, gclc, gclm, gss, trxt, trxr-1 and trxr-2) was evaluated via RT-PCR. Additionally, AChE activity was evaluated only after the 96h exposure period. The results indicated that Roundup exposure leads to a reduction in ROS levels in flies exposed for 96h. ACAP levels and gene expression of the antioxidant defence system exhibited an increase from 24h, while LPO did not show any significant alterations in both exposure periods. AChE activity was not affected following Roundup exposure. Our data suggest that Roundup exposure causes an early activation of the antioxidant defence system in D. melanogaster, and this can prevent subsequent damage caused by ROS.

A dissolution method for benzoyl metronidazole (BMZ) oral suspensions was developed and validated using a high-performance liquid chromatography (HPLC) method. After determination of sink conditions, dissolution profiles were evaluated using different dissolution media and agitation speeds. The sample insertion mode in dissolution media was also evaluated. The best conditions were obtained using a paddle, 50 rpm stirring speed, simulated gastric fluid (without pepsin) as the dissolution medium, and sample insertion by a syringe. These conditions were suitable for providing sink conditions and discriminatory power between different formulations. Through the tested conditions, the results can be considered specific, linear, precise, accurate, and robust. The dissolution profiles of five samples were compared using the similarity factor (f 2) and dissolution efficiency. The dissolution kinetics were evaluated and described by the Weibull model. Whereas there is no monograph for this pharmaceutical formulation, the dissolution method proposed can be considered suitable for quality control and dissolution profile comparison of different commercial formulations.