RESUMO
Depression is one of the world's most common and mentally disabling illnesses. Post-partum depression is a subtype of depression that affects one in seven women worldwide. Successful pharmacological treatment must consider the consequences for both, since the mother-child bond is fundamental for the well-being of both mother and infant as well as the general development of the newborn. Changes in maternal physiology and/or behavior can significantly influence the development of breastfed infants. Ketamine has been extensively studied for use as an antidepressant due to its mixed mechanisms of action. Safety and efficacy studies in the cardiovascular and urinary systems of a lactating postpartum depression animal model are essential for contributing toward ketamine's clinical use in the respective patient population. Thus, this project aimed to study the implications of postpartum maternal exposure to ketamine during lactation on the cardiovascular system of female rats submitted to the depression induction model by maternal separation. This model promotes depressive effects through stress caused by the interruption of mother-infant bond early in the offspring's life. To achieve depression, each dam was separated from her offspring for 3 h per day, from post-natal day 2 (PND2) to PND12. Experimental groups received daily treatment with either 5, 10, or 20 mg/kg of ketamine intraperitoneally during the lactation period, from PND2 to PND21. Behavioral tests consisted of the maternal and aggressive maternal behavior tests, the olfactory preference test, and the forced swim test. A technique for the detection of catecholamines and indoleamines in the heart muscle was developed for the experimental model groups. The histopathological evaluation was performed on these animals' cardiac muscles and urinary bladders. Our findings suggest that ketamine is safe for use in postpartum depression and does not induce cardiovascular and/or urinary systems toxicity.
RESUMO
Anticholinesterase pesticides are a main cause of the intentional or accidental poisoning of animals. Anticholinesterases include several substances that cause the overstimulation of both central and peripheral acetylcholine-dependent neurotransmission. Forensic analyses of poisoning cases require high levels of expertise, are costly, and often do not provide reliable quantitative information for unambiguous conclusions. The purpose of the present study was to develop and validate a method of high-performance liquid chromatography with diode array detector (HPLC−DAD) for the identification and quantitation of n-methyl carbamates, organophosphates and respective metabolites from biological samples of animals that were suspected of poisoning. HPLC−DAD is reliable, fast, simplistic and cost-effective. The method was validated for biological samples obtained from stomach contents, liver, vitreous humor and blood from four different animal species. The validation of the method was achieved using the following analytical parameters: linearity, precision, accuracy, selectivity, recovery, and matrix effect. The method showed linearity at the range of 25−500 µg/mL, and the correlation coefficient (r2) values were >0.99 for all matrices. Precision and accuracy were determined by the (a) coefficient of variation (CV), (b) relative standard deviation low-quality control (LQC), (c) medium-quality control (QCM), and (d) high-quality control (QCA). The indicated parameters were all less than 15%. The recovery of analytes ranged from 31 to 71%. The analysis of results showed no significant interfering peaks due to common xenobiotics or matrix effects. The abovementioned method was used to positively identify pesticide analytes in 44 of the 51 animal samples that were suspected of poisoning, demonstrating its usefulness as a forensic tool.
RESUMO
Objectives: The herbicide glyphosate, a pesticide used in agriculture to control weeds, both in food crops and in other agricultural areas, has been identified as an endocrine modulator through the inhibition of aromatase activity and the activation of estrogen receptors. The present study examined the effects of a glyphosate-based herbicide (Roundup® (GLY-BH) on sexual dimorphism of rats after perinatal exposure to low and high GLY-BH in males and females offspring. Methods: Two groups of pregnant rats were treated with two doses of GLY-BH (50 or 150 mg/kg) from day 15 of gestation (GD15) to postnatal day 7 (PND7). Play fighting behavior was observed at the juvenile stage and during social and sexual behaviors in adulthood. Results: Perinatal GLY-BH exposure reduced male and female body weight at 28, 75, and 90 days of age. The play fighting behavior was decreased in both sexes, but female rats were more affected. The sexual behaviors were reduced only in females. Conclusions: Perinatal exposure to both doses of GLY-BH promoted sexually dimorphic effects in both juvenile and adulthood stages. These effects were attributed to the inhibition of aromatase activity induced by exposure to GLY-BH in the perinatal period.(AU)
Objetivos: O glifosato é um herbicida não seletivo, usado em muitas culturas alimentares e não alimentares e em áreas não agrícolas, sendo que os produtos a base de glifosato atuam como moduladores das funções endócrinas por meio da inibição da atividade da aromatase e da ativação de receptores de estrógeno. O presente estudo avaliou os efeitos do herbicida Roundup® (GLY-BH) à base de glifosato, em comportamentos sexualmente dimórficos de ratos após exposição perinatal a doses baixas e altas de GLY-BH no período perinatal. Métodos: Ratas prenhas foram tratadas com 50 ou 150 mg/kg de GLY-BH do 15º dia de gestação (GD15) ao 7º dia de lactação (LD7). O comportamento de luta/brincar foi observado na fase juvenil e os comportamentos social e sexual na idade adulta. Resultados: a exposição perinatal a GLY-BH reduziu o peso corporal de machos e fêmeas aos 28, 75 e 90 dias de idade. O comportamento de luta/brincar diminuiu em ambos os sexos, sendo as ratas foram as mais afetadas. O comportamento sexual foi reduzido apenas nas fêmeas. Conclusões: A exposição perinatal a ambas as doses do GLY- BH promoveu tanto na idade juvenil como na idade adulta, efeitos sexualmente dimórficos. Esses efeitos foram atribuídos à inibição da atividade da aromatase induzida exposição perinatal ao GLY-BH.(AU)
Assuntos
Animais , Masculino , Feminino , Gravidez , Ratos , Comportamento Sexual Animal/fisiologia , Comportamento Social , Inibidores da Aromatase/efeitos adversos , Caracteres Sexuais , Herbicidas/administração & dosagem , Herbicidas/efeitos adversosRESUMO
BACKGROUND: More than 3 years since the last Zika virus (ZIKV) outbreak in Brazil, researchers are still deciphering the molecular mechanisms of neurovirulence and vertical transmission, as well as the best way to control spread of ZIKV, a flavivirus. The use of pesticides was the main strategy of mosquito control during the last ZIKV outbreak. METHODS: We used vesicular stomatitis virus (VSV) tagged with green fluorescent protein (GFP) as our prototypical virus to study the impact of insecticide pyriproxyfen (PPF). VZV-GFP infected and uninfected Jurkat, HeLa and trophoblast cells were treated with PPF and compared to untreated cells (control). Cell viability was determined by the MTT assay. Cell morphology, presence of extracellular vesicles (EVs), virus infection/GFP expression as well as active mitochondrial levels/localization were examined by confocal microscopy. RESULTS: PPF, which was used to control mosquito populations in Brazil prior to the ZIKV outbreak, enhances VSV replication and has cell membrane-altering properties in the presence of virus. PPF causes enhanced viral replication and formation of large EVs, loaded with virus as well as mitochondria. Treatment of trophoblasts or HeLa cells with increasing concentrations of PPF does not alter cell viability, however, it proportionately increases Jurkat cell viability. Increasing concentrations of PPF followed by VSV infection does not interfere with HeLa cell viability. Both Jurkats and trophoblasts show proportionately increased cell death with increased concentrations of PPF in the presence of virus. CONCLUSIONS: We hypothesize that PPF disrupts the lipid microenvironment of mammalian cells, thereby interfering with pathways of viral replication. PPF lowers viability of trophoblasts and Jurkats in the presence of VSV, implying that the combination renders immune system impairment in infected individuals as well as enhanced vulnerability of fetuses towards viral vertical transmission. We hypothesize that similar viruses such as ZIKV may be vertically transmitted via EV-to-cell contact when exposed to PPF, thereby bypassing immune detection. The impact of pesticides on viral replication must be fully investigated before large scale use in future outbreaks of mosquito borne viruses.
Assuntos
Infecções por Flavivirus/transmissão , Inseticidas/farmacologia , Piridinas/farmacologia , Vesiculovirus/efeitos dos fármacos , Replicação Viral/efeitos dos fármacos , Aedes/virologia , Animais , Brasil , Sobrevivência Celular/efeitos dos fármacos , Vírus da Dengue/efeitos dos fármacos , Vesículas Extracelulares/efeitos dos fármacos , Vesículas Extracelulares/virologia , Proteínas de Fluorescência Verde/genética , Células HeLa , Humanos , Células Jurkat , Trofoblastos/efeitos dos fármacos , Trofoblastos/virologia , Virulência , Zika virus/efeitos dos fármacosRESUMO
BACKGROUND: Depression is one of the most common mentally debilitating diseases in the world. Ketamine has been recently identified as a potential novel antidepressant. Further animal model evaluations of the use of ketamine as an antidepressant are necessary to determine safety parameters for clinical use. Therefore, the objective of this study was to perform toxicological tests of prolonged treatment using three different doses of ketamine in adult male rats. METHODS: The animals were divided into four groups: three treated with 5, 10 or 20 mg/kg of ketamine and a control group treated with saline solution. Intraperitoneal route of treatment was administered daily for 3 weeks. Body weight, water and food intake were measured once a week, as well as evaluation of the functional observational battery, which includes methodic monitoring of motor activity, motor coordination, behavioral changes, and sensory/motor reflex responses. Upon completion of treatment period, all animals were euthanized by decapitation followed by immediate collection of samples, which included brain structures and blood for neurochemical, hematological and biochemical analyses. RESULTS: Rats treated with the highest tested dosage (20 mg/kg) of ketamine had lower weight gain in the 1st and 2nd weeks of treatment and all experimental groups had measurable alterations in the serotoninergic system. CONCLUSIONS: Our data indicate that the alterations observed are minor and due to a predicted mechanism of action, which implies that ketamine is a promising drug for repurposing as an antidepressant.
Assuntos
Antidepressivos/administração & dosagem , Comportamento Animal/efeitos dos fármacos , Depressão/tratamento farmacológico , Ketamina/administração & dosagem , Animais , Antidepressivos/farmacologia , Antidepressivos/toxicidade , Depressão/fisiopatologia , Modelos Animais de Doenças , Relação Dose-Resposta a Droga , Ketamina/farmacologia , Ketamina/toxicidade , Masculino , Atividade Motora/efeitos dos fármacos , Ratos , Ratos Wistar , Testes de ToxicidadeRESUMO
Varenicline is a drug used for smoking addiction cessation treatment and acts as a partial agonist of nicotinic cholinergic receptors. Recent clinical trial data support use of varenicline for treatment of conditions/addictions that are not related to smoking cessation. Considering the importance of this issue and the need for new studies on its effects, especially on behavior, more studies using animal models are necessary. Thus, the aim of this study was to evaluate the effects of prolonged exposure to varenicline in anxiety-like behavior and memory, as well as in cerebral neurochemistry of rats. Male rats received three different doses of varenicline: 0.03 (therapeutic dose for humans), 0.1 and 0.3â¯mg/kg orally (gavage) for 30â¯days. Animal behavior was analyzed through open field, elevated plus maze, light/dark box, social interaction, Barnes maze and novel object recognition tests. Neurotransmitter levels and their metabolites in different brain structures (hippocampus, striatum and frontal cortex) were measured. Results showed that prolonged exposure of rats to varenicline: 1) did not interfere in motor activity, but caused an anxiogenic effect on elevated plus maze, light/dark box and social interaction testes; 2) did not alter memory; and 3) promoted alterations on serotoninergic system in the striatum and frontal cortex. In conclusion, compilation of the data indicates that prolonged exposure of rats to varenicline promoted anxiogenic effects and alteration in serotonergic system, which corroborated behavioral findings.
Assuntos
Ansiedade/induzido quimicamente , Memória/efeitos dos fármacos , Agonistas Nicotínicos/farmacologia , Neurônios Serotoninérgicos/efeitos dos fármacos , Vareniclina/farmacologia , Análise de Variância , Animais , Comportamento Animal/efeitos dos fármacos , Cognição/efeitos dos fármacos , Hipocampo/efeitos dos fármacos , Hipocampo/metabolismo , Masculino , Modelos Animais , Atividade Motora/efeitos dos fármacos , Nicotina/antagonistas & inibidores , Agonistas Nicotínicos/administração & dosagem , Ratos , Ratos Wistar , Serotonina/metabolismo , Fumar/tratamento farmacológico , Abandono do Hábito de Fumar/métodos , Vareniclina/administração & dosagem , Receptor Nicotínico de Acetilcolina alfa7/agonistas , Ácido gama-Aminobutírico/metabolismoRESUMO
Varenicline is a drug used for smoking addiction cessation treatment and acts as a partial agonist of nicotinic cholinergic receptors. Recent clinical trial data support use of varenicline for treatment of conditions/addictions that are not related to smoking cessation. Considering the importance of this issue and the need for new studies on its effects, especially on behavior, more studies using animal models are necessary. Thus, the aim of this study was to evaluate the effects of prolonged exposure to varenicline in anxiety-like behavior and memory, as well as in cerebral neurochemistry of rats. Male rats received three different doses of varenicline: 0.03 (therapeutic dose for humans), 0.1 and 0.3?mg/kg orally (gavage) for 30?days. Animal behavior was analyzed through open field, elevated plus maze, light/dark box, social interaction, Barnes maze and novel object recognition tests. Neurotransmitter levels and their metabolites in different brain structures (hippocampus, striatum and frontal cortex) were measured. Results showed that prolonged exposure of rats to varenicline: 1) did not interfere in motor activity, but caused an anxiogenic effect on elevated plus maze, light/dark box and social interaction testes; 2) did not alter memory; and 3) promoted alterations on serotoninergic system in the striatum and frontal cortex. In conclusion, compilation of the data indicates that prolonged exposure of rats to varenicline promoted anxiogenic effects and alteration in serotonergic system, which corroborated behavioral findings.
RESUMO
Varenicline is a drug used for smoking addiction cessation treatment and acts as a partial agonist of nicotinic cholinergic receptors. Recent clinical trial data support use of varenicline for treatment of conditions/addictions that are not related to smoking cessation. Considering the importance of this issue and the need for new studies on its effects, especially on behavior, more studies using animal models are necessary. Thus, the aim of this study was to evaluate the effects of prolonged exposure to varenicline in anxiety-like behavior and memory, as well as in cerebral neurochemistry of rats. Male rats received three different doses of varenicline: 0.03 (therapeutic dose for humans), 0.1 and 0.3?mg/kg orally (gavage) for 30?days. Animal behavior was analyzed through open field, elevated plus maze, light/dark box, social interaction, Barnes maze and novel object recognition tests. Neurotransmitter levels and their metabolites in different brain structures (hippocampus, striatum and frontal cortex) were measured. Results showed that prolonged exposure of rats to varenicline: 1) did not interfere in motor activity, but caused an anxiogenic effect on elevated plus maze, light/dark box and social interaction testes; 2) did not alter memory; and 3) promoted alterations on serotoninergic system in the striatum and frontal cortex. In conclusion, compilation of the data indicates that prolonged exposure of rats to varenicline promoted anxiogenic effects and alteration in serotonergic system, which corroborated behavioral findings.
RESUMO
Varenicline is a synthetic chemical substance produced from the alkaloid cytisine, used for smoking treatment, which acts as a partial agonist for α4ß2 and α3ß4 nicotinic cholinergic receptors and as a total agonist for α7 receptor. While there are studies regarding varenicline's non-smoking-related effects, as in treatment for drug dependence, there are no studies in the literature evaluating the long-term toxicity of varenicline through a physiological approach. Thus, the aim of this study was to evaluate possible toxicity through haematological, biochemical and anatomopathological parameters of prolonged exposure (30 days) to varenicline in rats. Three doses of varenicline were used: 0.03 (therapeutic dose for human beings), 0.1 and 0.3 mg/kg orally (gavage). Body-weight, water and food intake were measured weekly during treatment. On the 30th treatment day, blood and various organs were collected for haematological, biochemical and anatomopathological evaluations. The results show a decrease in some biochemical parameters in animals from the 0.1 and 0.3 mg/kg group, although the values are within the normal range of the species. There were no changes in the other evaluations performed. Together, these data indicate that prolonged exposure of rats to different doses of varenicline was not able to alter haematological, biochemical and anatomopathological parameters.
Assuntos
Agonistas Nicotínicos/efeitos adversos , Dispositivos para o Abandono do Uso de Tabaco/efeitos adversos , Vareniclina/efeitos adversos , Administração Oral , Animais , Relação Dose-Resposta a Droga , Ingestão de Líquidos/efeitos dos fármacos , Ingestão de Energia/efeitos dos fármacos , Coração/efeitos dos fármacos , Hematopoese/efeitos dos fármacos , Fígado/citologia , Fígado/efeitos dos fármacos , Fígado/metabolismo , Masculino , Miocárdio/citologia , Miocárdio/metabolismo , Agonistas Nicotínicos/administração & dosagem , Especificidade de Órgãos , Ratos Wistar , Reprodutibilidade dos Testes , Testes de Toxicidade Subaguda , Vareniclina/administração & dosagem , Aumento de Peso/efeitos dos fármacosRESUMO
O Paraquate (1,1'-dimetil-4,4'-bipiridina- dicloreto) é herbicida amplamente utilizado em vários países para diferentes culturas. O objetivo é determinar a concentração de Paraquate em batatas comercializadas em diferentes estabelecimentos da zona leste de São Paulo. Foram coletadas 12 (doze) amostras de batatas adquiridas no comércio varejista (sacolões, ou seja, do de frutas, verduras e legumes; supermercados e feiras livres) da zona leste do município de São Paulo. A quantificação do Paraquate foi baseada na reação de complexação com o ditionito de sódio, gerando composto de cor azulada, cuja absorvância foi lida em espectrofotômetro em comprimento de onda de 600nm. Foi construída a curva padrão e a determinada a equação da reta (y = 1,6448x e R2= 0,9945). O limite de tolerância do herbicida em alimentos é de 0,2 partes por milhão ou 0,2 mg/kg, enquanto que a ingestão diária aceitável (IDA) é de 0,004 mg/kg de peso corpóreo. Assim, pode-se observar que os valores encontrados em três amostras estão acima do limite máximo permitido, enquanto quatro apresentaram concentrações muito próximas ao limite. Os resultados permitem inferir que existe a necessidade de intensificação na fiscalização nos locais de comercialização de alimentos produzidos com a utilização de agrotóxicos.(AU)
Paraquat (1,1'-dimethyl-4,4'- bipyridine-dichloride) is herbicide widely used in several countries in different plantations. The objective is to determine the concentration of Paraquat in potatoes, marketed in different establishments in the eastern zone of São Paulo. Twelve (12) samples of potatoes purchased from the retail trade ("sacolões", ie fruit, vegetable and vegetable markets, supermarkets and free markets) were collected from the eastern part of the city of São Paulo. The quantification of Paraquat was based on the reaction of complexation with the sodium dithionite, generating compound of blue color, whose absorbance was read in a spectrophotometer at wavelength of 600 nm. The standard curve was constructed and the equation of the line was determined (y = 1,6448x e R2 = 0,9945). The tolerance limit of the herbicide in foods is 0.2 parts per million or 0.2 mg/kg, while the acceptable daily intake (ADI) is 0.004 mg/kg body weight. Thus, it can be observed that the values found in three samples are above the maximum allowed limit, while four of them presented concentrations very close to the limit. The results allow inferring that there is a need for intensification in the inspection in the commercial places of food produced with the use of pesticides.
Assuntos
Humanos , Paraquat/administração & dosagem , Paraquat/análise , Solanum tuberosum , Resíduos de Praguicidas , Amostras de Alimentos , Herbicidas/toxicidadeRESUMO
Agricultural pesticides used with the criminal intent to intoxicate domestic and wild animals are a serious concern in Veterinary Medicine. In order to identify the pesticide carbofuran and its metabolite 3- hydroxycarbofuran in animals suspected of exogenous intoxication a high pressure liquid chromatography with diode array detector (HPLC-DAD) method was developed and validated in stomach contents, liver, vitreous humor and blood. The method was evaluated using biological samples from seven different animal species. The following parameters of analytical validation were evaluated: linearity, precision, accuracy, selectivity, recovery and matrix effect. The method was linear at the range of 6.25-100µg/mL and the correlation coefficient (r2) values were >0.9811 for all matrices. The precision and accuracy of the method was determined by coefficient of variation (CV) and the relative standard deviation error (RSE), and both were less than 15%. Recovery ranged from 74.29 to 100.1% for carbofuran and from 64.72 to 100.61% for 3-hydroxycarbofuran. There were no significant interfering peaks or matrix effects. This method was suitable for detecting 25 positive cases for carbofuran amongst a total of 64 animal samples suspected of poisoning brought to the Toxicology Diagnostic Laboratory, School of Veterinary Medicine and Animal Sciences, University of Sao Paulo.
Assuntos
Carbofurano/análogos & derivados , Carbofurano/análise , Cromatografia Líquida de Alta Pressão/métodos , Medicina Legal/métodos , Intoxicação/diagnóstico , Intoxicação/veterinária , Medicina Veterinária/métodos , Animais , Carbofurano/sangue , Carbofurano/química , Gatos , Cães , Conteúdo Gastrointestinal/química , Limite de Detecção , Modelos Lineares , Fígado/química , Reprodutibilidade dos Testes , Corpo Vítreo/químicaRESUMO
Animal cruelty is a known behavior of psychopaths, and although the serial killing of humans is widely acknowledged worldwide, this type of crime against animals is seldom discussed. This report describes the necropsy and toxicological findings of 37 dogs and cats, which were found dead in plastic bags in Sao Paulo, Brazil. The animals had all been in the care of an alleged animal rescuer and were to be referred for adoption before being found dead. In the necropsy, the animals showed varying degrees of putrefaction, indicating different periods of death, as well as single or multiple perforations on the thorax. The perforations reached the heart, lungs or large thoracic vessels, culminating in hemopericardium and hemothorax that led to death by circulatory failure and cardiac tamponade. Blood from the heart and thoracic cavity was analyzed by gas chromatography coupled with mass spectrometry (GC-MS) and tested positive for ketamine, a dissociative anesthetic. The suspect declared that she had killed only five of the animals and that they had all been fatally sick. The necropsy proved that all 37 animals were killed in the same way, that none of the animals had any terminal diseases and that a restricted drug was used. The suspect was sentenced to 12 years, 6 months and 14days of prison for the killing of the 37 animals. This was the first conviction for the crime of animal cruelty in Brazil. The combined role of police, forensic veterinary pathologists and prosecutors were essential to the conviction, which was a great historical occasion in the fight against animal cruelty.
Assuntos
Anestésicos Dissociativos/sangue , Bem-Estar do Animal/legislação & jurisprudência , Crime , Ketamina/sangue , Animais , Brasil , Gatos , Cães , Feminino , Cromatografia Gasosa-Espectrometria de Massas , HumanosRESUMO
Exhumation is required for the investigation of suspicions deaths when a body is buried and is usually performed under court order. Exhumation of animals is not a routine practice in forensic pathology. In this study, 30 male 70-day-old Wistar rats were experimentally exposed to the carbamate pesticides aldicarb and carbofuran. Toxicological, macroscopic and microscopic examinations were performed. Groups of 3 animals (2 exposed and 1 control) were evaluated at 24h, 3days, 5days, 7days and 10days post-mortem. In histopathological examination, the brain, liver, lungs and kidneys were assessed, and for toxicological analysis, the gastric contents, liver, vitreous humor, skeletal muscle and larvae (when available) were collected. The pesticides were detected by HPLC and quantified in the analyzed matrices, and a possible delay in tissue putrefaction due to the pesticides was observed. This study has revealed that it is possible to exhume animals for investigations of possible poisoning by carbamates and has demonstrated that the exhumation of an animal in a suspected case of poisoning should not be ruled out. The increasing demand for investigations of suspicious animal deaths, e.g., in cases of poisoning, will likely lead to an increase in the use of this type of procedure in veterinary pathology.
Assuntos
Aldicarb/intoxicação , Carbofurano/intoxicação , Exumação/métodos , Inseticidas/intoxicação , Síndromes Neurotóxicas/veterinária , Aldicarb/análise , Animais , Carbofurano/análise , Cromatografia Líquida de Alta Pressão , Inseticidas/análise , Masculino , Ratos , Ratos WistarRESUMO
The intentional and accidental poisoning of animals and people is a threat to public health and safety worldwide. Necropsies and histopathological examinations of 26 cats and 10 dogs poisoned by the carbamates aldicarb and carbofuran, confirmed by thin layer chromatography (TLC) and high performance liquid chromatography with diode-array detector (HPLC-DAD) were analysed, with variable post mortem interval and conservation of the carcass. Biological matrices were collected for toxicological and histopathological analyses. High performance liquid chromatography with diode-array detector (HPLC-DAD) was utilized to detect aldicarb and its metabolites, aldicarb sulphoxide and aldicarb sulphone, and carbofuran. The variable post mortem interval and the method of conservation of the carcass may be harmful to toxicological, necroscopic and histopathological analyses, that should be performed in order to provide reliable evidences to investigate possible poisoning of animals, which is cruel crime, and are usually linked to domestic or social conflict.