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Metalworkers are exposed to numerous chemicals in their workplace environment, such as solvents, heavy metals, and metalworking fluids, that have a negative impact on their health. Furthermore, there is an increase in the prevalence of chronic diseases among metalworkers; however, the molecular mechanisms involved in this increased predisposition to chronic diseases are unclear. Considering that occupational exposure represents a potential risk for metalworkers, the aim of this study was to measure biomarkers of oxidative stress, inflammation, and cytotoxicity in the peripheral blood of metalworkers from Southern Brazil. The study included 40 metalworkers and 20 individuals who did not perform activities with any recognized exposure to chemical substances, such as those working in administration, commerce, and education, as controls. Cellular and molecular biomarkers as leukocyte viability, intracellular production of reactive species, mitochondrial mass and membrane potential and plasma lipid peroxidation, sulfhydryl groups, total antioxidant capacity, and butyrylcholinesterase activity were evaluated in the blood of metalworkers and controls. Metalworkers were found to have higher rates of apoptosis, increased production of reactive species, and increased mitochondrial potential and mass in leukocytes associated with decreased antioxidant defenses and increased activity of the butyrylcholinesterase enzyme in their plasma. It can be concluded that cytotoxicity, oxidative stress, and inflammation are involved in the multiplicity of health outcomes related to chemical exposure in the metalworking industry.

O mastocitoma cutâneo e uma neoplasia originada a partir da proliferacao desordenada de mastocitos, sendo altamente infiltrativo no subcutâneo e na derme. Em gatos representa 12 a 20% dos tumores de pele. No entanto, relatos de casos desse tumor no Brasil sao raros. O objetivo deste trabalho e descrever aspectos clinicos, citologicos e hematologicos de mastocitoma cutâneo em um felino femea, sem raca definida, pesando 2,7 kg, com 3 anos de idade, atendido no Hospital Veterinario da Universidade de Cruz Alta com historico de que a cerca de 6 meses apresentou aumento de volume progressivo acima do olho direito. Ao exame fisico foi evidenciado mucosas coradas, temperatura normal, tempo de preenchimento capilar < 2 segundos e nodulo solitario, bem circunscrito, nao ulcerado, de consistencia firme e de aproximadamente 5 cm na face. Apos a realizacao de citologia aspirativa por agulha fina foi obtido o diagnostico de mastocitoma cutâneo. A partir da citologia esse tumor e classificado na categoria de neoplasias de celulas redondas, juntamente com o linfoma, tumor venereo transmissivel, entre outros, por isso estes devem ser inclusos como diagnostico diferencial. Nesse contexto, os exames laboratoriais desempenham importante papel no diagnostico da doenca e monitoramento da saude do paciente.

O Tumor Venereo Transmissivel (TVT) apresenta maior prevalencia em caes jovens, sem predilecao por raca e sexo, ocorrendo com mais frequencia em regioes onde os caes circulam livremente. Surge como uma massa irregular hiperemica e sanguinolenta, transmitido por meio da transferencia muco-cutânea de celulas neoplasicas atraves do contato durante o coito ou por habitos de cheirar ou lamber. Essa condicao pode acarretar aos animais diminuicao da qualidade de vida devido a dor em regioes acometidas, secrecao vaginal ou peniana continuas, prurido intenso, mudanca de comportamento visualizados por apatia, anorexia e letargia, alem de se tornarem os principais reservatorios dessa neoplasia podendo transmitir a outros animais. Apos o diagnostico conclusivo que pode ser realizado atraves da citologia, o tratamento quimioterapico com vincristina tem se mostrado eficaz e possibilita a remissao da massa neoplasica. Objetivando analisar a prevalencia do TVT e sua distribuicao quanto ao sexo, idade, raca, area corporal acometida e formas de coleta das amostras para diagnostico, avaliou-se 89 caninos atendidos no Hospital Veterinario da Unicruz no periodo de janeiro de 2010 a janeiro de 2020. Com base nos dados obtidos concluiu-se que a maioria dos caninos diagnosticados com TVT na regiao de Cruz Alta, sao femeas (69,31%), em idade reprodutiva de 1 a 8 anos (30,68%), sem raca definida (80,68%) e com maior numero de lesoes observadas em regiao genital (73,87%). A tecnica de diagnostico mais utilizada na rotina clinica foi o imprint (72,72%), seguido da citologia aspirativa por agulha fina (14,77%), swab (4,54%) e imprint associado a citologia (4,54%). Com isso destaca-se a importância desse estudo retrospectivo realizado na regiao de Cruz Alta e cidades vizinhas, a fim de possibilitar o estudo de dados importantes sobre a epidemiologia dessa neoplasia, formas de coleta de amostras para analise e da possibilidade de realizacao do diagnostico definitivo a partir de interpretacao citologica de baixo custo.

Endometritis is an inflammation of the endometrium associated with bacterial infection. The pathogenesis of endometritis in cows is still not completely understood. The combined analysis of the markers of inflammation and oxidative stress has contributed to a better understanding of disease mechanisms, but is still unexplored in uterine disorders. Moreover, research provides evidence about an important role of the vagus nerve in regulating the innate immune function through the cholinergic anti-inflammatory pathway in response to bacterial infections. This new pathway has demonstrated a critical role in controlling the inflammatory system. The aim of this study was to evaluate the activity of cholinesterase in total blood, lymphocytes, and serum of dairy cows with clinical and subclinical endometritis. Sixty-one Holstein cows, between 30 and 45 days in milk, were classified into 3 groups of animals: presenting clinical endometritis (n = 22), subclinical endometritis (n = 17), and healthy (n = 22). Mean leukocyte counts did not differ among groups, but the neutrophil number was significantly higher in cows with clinical endometritis than those in healthy animals. Also, serum concentration of interleukin-1beta (pg/mL) was significantly higher in cows with endometritis. The activity of acetylcholinesterase in blood and lymphocytes increased in both groups with endometritis. Animals with endometritis presented an increase in lipid peroxidation, but the antioxidant enzyme activity (catalase levels) was higher in endometritis groups than in normal cows. In conclusion, the inflammatory process of clinical and subclinical endometritis leads to systemic lipid peroxidation despite the compensatory increase of the antioxidant enzyme. These data also provide evidence of an important role of the cholinergic pathway in regulating dairy cows with clinical and subclinical endometritis.

The present study aimed to evaluate the effects of a strategic feeding supplementation protocol on reproductive and economic results of exclusively fixed timed artificial inseminated (FTAI) beef cows from a southern Brazilian herd. The experiment was carried out over 2 consecutive breeding seasons (2016 and 2017, from October to January). Two-hundred, 3 and 4 years old, non-suckled, Angus cows (n=100/year), were divided into two homogeneous groups (by weight and body condition score) 25 days before the first FTAI of the breeding season (First FTAI=Day 0). As a representation of traditional management of herds grazing on natural pastures from Rio Grande do Sul Province, Brazil, cows from control group (CG; n=100; 50 cows in each year) received basic mineral supplementation (without protein an energy) ad libitum during the entire experimental period. As an alternative feeding protocol, supplemented group (SG; n=100; 50 in each year) received a mineral supplementation enriched with protein (23%), energy (44% NDT) and sodium monensin (0.25%) ad libitum from days: –25 to 80 of breeding season. Cows were maintained in 2 separated paddocks of native pasture with similar forage composition and availability. All cows were submitted to a progesterone/estradiol-based estrus synchronization protocol on day -10, and cows not pregnant at diagnosis were resynchronized on days 28 and 76 using the same hormonal treatment. There was no year effect (P>0.1) on weight gain and reproductive results, data from both breeding seasons were polled together for further analyzes. Cows from SG presented higher average daily weight gain and gained more weight than cows from CG (p0.1). Cows from SG became pregnant earlier (p

Durante a exposição a agrotóxicos ocorrem alterações fisiológicas no organismo humano, como o estresse oxidativo, caracterizado por um desequilíbrio entre o sistema de defesa antioxidante e a geração de espécies reativas. Os agrotóxicos podem ocasionar alterações inflamatórias, em algumas citocinas, que são essenciais para o funcionamento do nosso organismo. Neste contexto, o objetivo desse estudo foi avaliar o perfil oxidativo e inflamatório de agricultores expostos a agrotóxicos. Foram utilizadas amostras de sangue de 50 agricultores e 50 indivíduos não expostos a agrotóxicos (Grupo Controle – GC). Foram mensurados no plasma dos participantes marcadores de dano oxidativo e marcadores inflamatórios. Foi encontrada uma elevação nos níveis de TBARS (p

The aim of this study was to evaluate whether oxidative stress occurs in rats experimentally infected by Sporothrix schenckii, and its possible effect on disease pathogenesis. Thirty rats were divided into two groups: the group A (uninfected, n = 18) and the group B (infected by S. schenckii, n=21). Blood samples were collected on days 15, 30 and 40 post-infection (PI). At each sampling time, six rats of the group A, and seven of the group B were bled. TBARS (thiobarbituric acid reactive substances) levels in serum samples were measured to evaluate lipid peroxidation. In addition, catalase (CAT) and superoxide dismutase (SOD) activities, known as biomarkers of antioxidants levels, were verified in whole blood. Seric pro-inflammatory cytokine levels were measured (IFN-γ, TNF-α, and IL-6), which showed that these inflammatory mediators were at higher levels in the infected rats (P

In healthy cartilage, chondrocytes maintain an expression of collagens and proteoglycans and are sensitive to growth factors and cytokines that either enhance or reduce type II collagen synthesis. In osteoarthritis, pro-inflammatory cytokines, such as IL-6, induce overexpression of metalloproteinases (MMP) and decreasing synthesis of aggrecan. Use of chondroprotectors agents, such as Platelet-Rich Plasma (PRP) and triamcinolone (TA) are alternatives to reduce the progression of joint damage. In this study, we used chondrocytes extracted from metacarpophalangeal joints of healthy horses as the experimental model. Cells were treated in vitro with PRP or TA. No differences were observed between these treatments in comparison to the control group when the expressions of MMP9, MMP13, IL-6 and ACAN genes were evaluated (P

Bovine herpesvirus type 5 (BoHV-5) is the causative agent of herpetic meningoencephalitis in cattle. The purinergic system is described as a modulator of the immune response and neuroinflammation. These functions are related to the extracellular nucleotides concentration. NTPDase and 5′-nucleotidase are enzymes responsible for controlling the extracellular concentration of adenosine triphosphate (ATP), adenosine diphosphate (ADP), adenosine monophosphate (AMP), and adenosine (ADO). The aim of this study is to determinate the ectonucleotidase activity in cortical synaptosomes and synaptosomes from the hippocampus of rabbits experimentally infected with BoHV-5. Rabbits were divided into four groups, two control groups (non-inoculated animals), and two infected groups (inoculated with BoHV-5). The infected groups received 0.5 ml of BoHV-5 suspension with 107.5TCID50 of viral strain SV-507/99, per paranasal sinuses, and the control groups received 0.5 ml of minimum essential media per paranasal sinuses. Animals were submitted to euthanasia on days 7 and 12 post-inoculation (p.i.); cerebral cortex and hippocampus were collected for the synaptosomes isolation and posterior determination of the ectonucleotidase activities. The results showed a decrease (P

The aim of this study was to analyze the classic iron markers associated to the storage process in hamsters experimentally infected by Leptospira interrogans serovar Pomona. Four groups with six hamsters each were used; two were negative controls (C7 and C14) and two were composed by infected animals (T7 and T14). Blood samples were collected on the seventh (C7 and T7) and fourteenth days (C14 and T14) post-inoculation. Iron availability was determined in sera samples through the assessment of iron, ferritin, transferrin, and iron binding capacity, whereas the bone marrow was also evaluated for the presence of iron by Pearl's reaction. Additionally, the total antioxidant capacity (TAC) and total oxidant status (TOS) were assessed, along with hepcidin and IL-6 levels. Based on the results, it was possible to observe the onset of an anemic profile, predominantly hemolytic and regenerative. Also, The other parameters showed an increase in seric iron (P

Objetivou-se comparar a resposta inflamatória e o perfil oxidativo da técnica de ovariossalpingohisterectomia (OSH) convencional com duas técnicas de Cirurgia Endoscópica por Orifícios Naturais (NOTES). Foram utilizados 15 fêmeas, alocadas em três grupos de cinco animais. No primeiro grupo, a OSH foi realizada por celiotomia convencional, no segundo, por NOTES total e, no terceiro, por NOTES híbrida. Foram realizadas as coletas sanguíneas antes do procedimento cirúrgico (basal), 3, 6, 12, 24, 48 e 72h pós-operatórias. A atividade da catalase manteve-se alta nos três grupos estudados, entretanto a peroxidação lipídica, medida pelos níveis dos produtos de reação com o ácido tiobarbitúrico (TBARS), ocorreu mais acentuadamente no grupo convencional e foi quase que imperceptível no grupo de NOTES total. Nos três grupos estudados, ocorreu elevação na atividade da butirilcolinesterase e acetilcolinesterases, bem como aumento leucocitário neutrofílico nas primeiras horas pós-cirúrgicas. Conclui-se que a inflamação sistêmica acontece de forma similar nas três técnicas operatórias, com ressalva para as realizadas por NOTES total que mantiveram as mais baixas taxas oxidativas. This study aimed at comparing the inflammatory response and the oxydative profile of the conventional ovarysalpingohysterectomy (OSH) technique to Totally Natural Orifice Transluminal Endoscopic Surgery (NOTES) and Hybrid NOTES. Group of fifteen female dogs was used for each technique. Blood samples were taken before the surgical procedure (basal) and 3, 6, 12, 24, 48 and 72h postoperative. The catalase activity was increased in the three studied groups. Nevertheless, lipid peroxidation, measured by TBARS (thiobarbituric acid reactive substances) levels, was higher in the conventional group and almost indistinguishable in the total NOTES group. In the three analyzed groups, both butyrylcholinesterase and acetylcholinesterase activities were increased as well as the neutrophil counts during the first post-surgical hours. It is possible to conclude that systemic inflammation occurs in a similar way in the three operative techniques; however, total NOTES technique presents lower levels of cellular oxidative damage, particularly if compared to the conventional approach.

This experiment aims to evaluate the correlation between lipid peroxidation levels in serum and seminal plasma in equines. Also, it investigates the lipid peroxidation in extended semen samples and its effects and sperm motility during a 72 hr refrigeration period. Blood and semen were collected from fertile Crioulo stallions. Serum and seminal plasma lipid peroxidation levels were analyzed by thiobarbituric acid reactive substances (TBARS) immediately after semen collection. After addition of extender (hour = 0), diluted semen was refrigerated and stored at 5 °C. Semen analyses, TBARS and catalase activity were performed in extended semen at 0, 24, 48, and 72 hours. We noted that levels of plasma lipid peroxidation can be used as an indicative of seminal oxidative stress. Also, lipid peroxidation does not increase substantially during semen storage. Lipid peroxidation and the antioxidant enzyme catalase do not seem to be the major cause of loss and motility and consequently reduction in fertility in stallion semen during storage for 72 h at 5 °C.

This study evaluated the effects of curcumin and/or insulin on antioxidant enzyme activity in blood, liver, and kidney, as well as on lipid peroxidation and delta aminolevulinic dehydratase (δ-ALA-D) activity, and a histopathological analysis of streptozotocin-induced diabetic rats. The animals were divided into six groups (n = 6): control/saline (C); control/curcumin (CCur); diabetic/saline (D); diabetic/insulin (DIns); diabetic/curcumin (DCur); and diabetic/insulin/curcumin (DInsCur). After 30 days of treatment with curcumin and/or insulin, the animals were sacrificed and the liver, kidney, and serum were used for experimental determinations. Results of histopathological analysis showed that the treatment with insulin ameliorate renal and hepatic lesions from both DIns and DInsCur groups. TBARS levels were significantly increased in serum, liver, and kidney in D group and the administration of curcumin and insulin prevented this increase in DIns and DCur groups. The activities of catalase (CAT), superoxide dismutase, and δ-ALA-D presented a significant decrease in the liver and kidney D group when compared to C group (P < 0.05). The animals treated with curcumin and insulin presented an increase of CAT activity, revealing a positive interaction between both substances. The treatments with curcumin or insulin prevented oxidative stress in blood, through modulation of enzymatic antioxidant defenses. These findings contributed to the comprehension that antioxidants from medicinal plants could be used as adjuvant in the treatment of this endocrinopathy and not as single therapy.

SUMMARYThe aim of this study was to investigate neurochemical and enzymatic changes in rats infected withTrypanosoma evansi, and their interference in the cognitive parameters. Behavioural assessment (assessment of cognitive performance), evaluation of cerebral L-[3H]glutamate uptake, acetylcholinesterase (AChE) activity and Ca+2and Na+, K+-ATPase activity were evaluated at 5 and 30 days post infection (dpi). This study demonstrates a cognitive impairment in rats infected withT. evansi. At 5 dpi memory deficit was demonstrated by an inhibitory avoidance test. With the chronicity of the disease (30 dpi) animals showed anxiety symptoms. It is possible the inhibition of cerebral Na+, K+-ATPase activity, AChE and synaptosomal glutamate uptake are involved in cognitive impairment in infected rats byT. evansi. The understanding of cerebral host–parasite relationship may shed some light on the cryptic symptoms of animals and possibly human infection where patients often present with other central nervous system (CNS) disorders.

The potent activity against Trypanosomes and health beneficial effects of curcumin (Cur) has been demonstrated in various experimental models. In this study, we evaluated the in vivo effect of Cur as trypanocide and as potential anti-inflammatory agent, through the evaluation of immunomodulatory mechanisms in rats infected with Trypanosoma evansi. Daily oral Cur was administered at doses of 0, 20 or 60mg/kg as preventive treatment (30 and 15days pre infection) and as treatment (post infection). The treatment of the groups continued until the day of euthanasia. Fifteen days after inoculation, parasitemia, plasma proinflammatory cytokines (IFN-γ, TNF-α, IL-1, IL-6), anti-inflammatory cytokines (IL-10) and blood acetylcholinesterase activity (AChE) were analyzed. Pretreatment with Cur reduced parasitemia and lethality. Cur inhibited AChE activity and improved immunological response by cytokines proinflammatory, fundamental during T. evansi infection. We found that Cur is not so important as an antitrypanosomal activity but as immunomodulator agent. These findings reveal that the preventive use of Cur stimulates anti-inflammatory mechanisms, reducing an excessive inflammatory response.

Pythium insidiosum causes life-threatening disease in mammals. Animals with pythiosis usually develop anemia, and most human patients are reported to have thalassemia and the major consequence of thalassemia, iron overload. Therefore, this study evaluated the iron metabolism in rabbits experimentally infected with P. insidiosum. Ten infected rabbits were divided into two groups: one groups received a placebo, and the other was treated with immunotherapy. Five rabbits were used as negative controls. The hematological and biochemical parameters, including the iron profile, were evaluated. Microcytic hypochromic anemia was observed in the infected animals, and this condition was more accentuated in the untreated group. The serum iron level was decreased, whereas the transferrin level was increased, resulting in low saturation. The level of stainable iron in hepatocytes was markedly decreased in the untreated group. A high correlation was observed between the total iron binding capacity and the lesion size, and this correlation likely confirms the affinity of P. insidiosum for iron. The data from this study corroborate the previous implications of iron in the pathogenesis of pythiosis in humans and animals.

Objectives: Iron plays an important role in the pathogenesis of Pythium insidiosum. Human pythiosis frequently occurs in iron-overloaded thalassaemic patients and experimentally infected animals develop iron deficiency anaemia. Therefore, we sought to determine the in vitro and in vivo activities of the iron chelator deferasirox against P. insidiosum. Methods: In vitro, the MIC and minimum fungicidal concentration (MFC) values of deferasirox for 17 strains of P. insidiosum were determined in accordance with CLSI document M38-A2. In vivo studies were carried out in 20 inoculated rabbits divided into four groups: placebo, immunotherapy obtained from vortexed P. insidiosum cultures (14 day intervals), deferasirox (15 mg/kg/day) and a combination of immunotherapy and deferasirox. Five non-infected animals were used as controls. Results: The MIC and MFC values of deferasirox for P. insidiosum ranged from 12.5 to 50 mg/L and from 50 to 100 mg/L, respectively. Treatment with deferasirox alone ameliorated anaemia and normalized the serum iron levels and hepatic iron concentration in the animals. However, the mean lesion size, although decreased, did not differ significantly from that in the placebo group. The results of immunotherapy plus iron chelation therapy were worse than those of immunotherapy alone. Moreover, the disease spread to the lung tissue in 5 out of 10 deferasirox-treated animals. Conclusions: Despite its limited in vitro and in vivo activity, deferasirox improved iron deficiency anaemia in P. insidiosum-infected rabbits. Further studies are needed to investigate the immunomodulatory properties observed in this study and the benefits and drawbacks of using iron-chelating drugs as an adjuvant therapy in pythiosis.

Several chemical and immunohistochemical techniques can be used for the detection of acetylcholinesterase (AChE) activity. In this experiment we aimed to detect AChE activity in Trypanosoma evansi. For this, the parasites were isolated from the blood of experimentally infected rats using a DEA-cellulose column. Enzymatic activity was determined in trypomastigote forms at 0, 0.2, 0.4, 0.8 and 1.2mg/mL of protein concentrations by a standard biochemical protocol. At all concentrations tested, the study showed that T. evansi expresses the enzyme AChE and its activity was proportional to the concentration of protein, ranging between 0.64 and 2.70μmol of AcSCh/h. Therefore, we concluded that it is possible to biochemically detect AChE in T. evansi, an enzyme that may be associated with vital functions of the parasite and also can be related to chemotherapy treatments, as further discussed in this article.

Leptospirosis is a systemic disease of humans and domestic animals, mainly dogs, cattle and swine. The course of human leptospirosis varies from mild to severe fatal forms and the most severe form of human leptospirosis is principally caused by Leptospira interrogans serovar icterohaemorrhagiae (L. icterohaemorrhagiae). The enzyme adenosine deaminase (ADA) plays an important role in the production and differentiation of blood cells. The aim of this study was to evaluate the activity of ADA in serum, erythrocytes and lymphocytes of rats infected with L. icterohaemorrhagiae, as compared with non-infected rats. Twenty-four adult rats, divided into two uniform groups (A and B) were used for the enzymatic assays. The animals in Group B were inoculated intraperitoneally with 2×10(8) leptospires/rat, and the rodents in Group A (control) were not-inoculated. Blood collection was performed on days 5 and 15 post-infection (PI) and the blood used to assess the ADA activity. The infection by L.icterohaemorrhagiae altered erythrocyte count, hemoglobin concentration and hematocrit, causing a decrease in all these parameters on day 15 PI. Lymphocytes decreased significantly on day 15 PI, and ADA activity in serum was inhibited in infected rats on days 5 and 15 PI and its activity in erythrocytes were increased on day 5 PI. On day 5 PI, we found an increase in ADA activity in erythrocytes of infected rats. No correlation was observed between hematocrit and erythrocyte ADA activity on days 5 and 15 PI. The ADA activity was inhibited in rats infected on day 15 PI. A positive correlation (r(2)=60) was also observed between the number of lymphocytes and ADA activity in lymphocytes on day 15 PI (P0.05). In conclusion, our results showed that the ADA activity is altered in serum, lymphocytes and erythrocytes in experimental infection by L.icterohaemorrhagiae in rats, concomitantly with hematological parameters.

This study aimed to assess plasma concentrations of copper, iron, and zinc during the course of acute Trypanosoma evansi experimental infection in rats. Fifty male Wistar rats were randomly distributed into seven groups: three trypanosome-infected groups (T2, T4, and T6; n = 10 animals per group) and four uninfected controls (C0, C2, C4, and C6; n = 5 animals per group). Animals from trypanosome-infected groups were intraperitoneally injected with 106 trypanosomes/animal. Blood samples were collected by cardiac puncture before infection (day 0; group C0) or on the second (C2 and T2), fourth (C4 and T4), and sixth (C6 and T6) day postinfection. Parasitemia and hematological evaluation were performed to assess the progression of the disease in animals. The difference between groups (control and infected) was evaluated on the same day postinfection. Plasma copper concentration increased in T4 and T6 groups (P < 0.001) compared with the control group. Plasma iron concentration decreased only in group T2 (P < 0.001). There was no significant difference in the plasma zinc concentration between groups. This study therefore demonstrates that high plasma copper concentration and depression in iron concentration is part of the acute phase response in rats infected with T. evansi.

Pyometra is a common uterine disease found in adult female dogs during diestrus and it typically presents with several systemic abnormalities. The aim of this study was to evaluate complete blood counts (CBC), neutrophil oxidative metabolism, and lipid peroxidation in female dogs diagnosed with pyometra associated with Escherichia coli (E. coli). Twenty adult female dogs, from 2 to 14 years old, were allocated into two different groups. Animals placed in group 1 (G1) (n = 12) were diagnosed with pyometra associated with E. coli and animals placed in group 2 (G2) (n = 8) were clinically healthy female dogs. Red blood cell (RBC) count, packed cell volume, and hemoglobin were significantly lower in G1 than in G2. A significant increase in total plasma protein and white blood cell count (segmented neutrophils, band neutrophils, and monocytes) occurred in G1 when compared to G2. The neutrophil oxidative metabolism was also increased in G1, but lipid peroxidation showed no difference between the two groups. CBC results revealed a normocytic, normochromic anemia and leukocytosis characterized by a neutrophilia with a left shift in adult female dogs with pyometra associated with E. coli. Additionally, neutrophil oxidative metabolism showed an increase in its activity without producing lipid peroxidation, which possibly indicates that the anemia in canine pyometra is not related to oxidative stress.

The present study evaluates levels of aflatoxin contamination involving small dairy farms from the in the Northwest region of Rio Grande do Sul, Brazil. The objectives of this trial is: quantify lactating cow diet and raw for bulk milk Aflatoxin B1 (AFB1) contamination; evaluate the use of a mycotoxin adsorbent on cows submitted to semi-confinement raising; and verify the presence of AFM1 contamination on UHT and powdered milk commercialized locally. For this, 17 farms were selected. During the experimental period, every lactating cow on each herd received 50 g/day of a commercial mycotoxin adsorbent for dairy cows (100% bentonite). From each farm, bulk milk, corn silage and concentrate samples were collected for aflatoxin quantification. Aflatoxin M1 was detected in all milk samples examined from each farm, but the levels were within the allowed by the legislation. Aflatoxin B1 was also detected in all samples of corn silage and concentrate. The mycotoxin adsorbent used reduced (p < 0.05) AFM1 contamination in milk. AFM1 milk contamination levels observed on commercialized milk also respected Brazilian requirements. It is possible to conclude that there is a high incidence of Aflatoxin B1 contamination on corn silage and dairy cows concentrates in family farms from South Brazil region. Based on the condition in which the present study was conducted, the use of a mycotoxin adsorbent reduced the levels of AFM in milk in production system. However, the locally produced milk and commercial milk were below the AFM levels accepted by the National Requirements for AFM1 contamination.

Background : : : : Trypanosoma evansi ( T. evansi ) is a protozoan which causes trypanosomosis in livestock in many countries of Southeast Asia, Africa and South America. Patterns of disease vary from acute epidemics with high case-fatality rates to subclinical and/or chronic disease in endemic animal populations. It is a problem of great economic importance due to the death of sick animals and high cost of treatment. This article aims to review the outbreaks of the infection by T. evansi in horses that occurred in southern Brazil. Review : These outbreaks were discussed in terms of epidemiology, clinical signs, laboratory tests, pathological findings, diagnosis and treatment by addressing the differences between the cases occurred in the state of Rio Grande do Sul and in other Brazilian states. The outbreaks due to T. evansi in livestock animals are endemic in warm-climate areas. At the Rio Grande do Sul state, most of the equine trypanosomosis occurs in the summer. This can be easily explained by the high number of bloodsucking insects, which are responsible for the mechanical transmission of the flagellate among the animals. Clinical signs such as progressive weight loss, lethargy, incoordination, instability, atrophy and paralysis of the hind limbs, difficulty in standing and walking, subcutaneous edema and abortion are often reported in T. evansi- infected equines. Anemia is the clinical alteration most observed in these infections, although its pathogenicity still remains unclear. In the present study anemia was associated with lipid peroxidation and decrease in serum iron levels and in acetylcholinesterase activity. Necropsy alterations found in the outbreaks reported in the Rio Grande do Sul state are commonly described in infections by T. evansi , except by the neurological alterations as necrotizing encephalitis of the white matter associated with edema, demyelinization and perivascular lymphoplasmocytic infiltrate. The diagnosis of the equine trypanosomosis was based on morphology and biometry of the trypomastigote forms in peripheral blood smears stained with Quick Panoptic or Giemsa methods, immunohistochemic, xenodiagnostic, and PCR T. evansi- specific. A new therapeutic protocol using diminazene aceturate at a dose of 7 mg kg -1 was tested in one of the outbreaks. This approach cured all the infected animals. Conclusion : Although T. evansi was diagnosed for the first time in Rio Grande do Sul state in 2002, veterinary clinicians have reported clinical signs such as paralysis of the pelvic members, fever and weight loss since the 80’s and 90’s. Therefore, the lack of knowledge of the disease might have been responsible for the unpublished data. Another hypothesis is the suspect of babesiosis, since both illnesses have marked anemia and hyperthermia. Moreover, as the diminazene aceturate has trypanocidal and babesicidal action, animals may have been misdiagnosed with babesiosis and may have recovered from the T. evansi infection. Only few researches on trypanosomosis are found in the southern region of Brazil. Prevalence studies with more sensible techniques are necessary in order to clarify the spread of the disease and the economic losses that it causes to farmers.

The aim of this study was to evaluate cholinesterase activity during the early acute phase of Trypanosoma evansi infection in rats. Fifteen male Wistar rats were randomly distributed into three groups ( n = 5 animals per group): two trypanosome-infected groups (T3 and T5) and uninfected controls (C). The animals were inoculated intraperitoneally with 10 6 trypanosomes. The blood was collected by cardiac puncture on the 3rd (T3) or 5th day post-infection (T5 and C). Cerebrum and cerebellum were removed for the evaluation of acetylcholinesterase (AChE) activity. AChE activity was also evaluated in whole blood and butyrylcholinesterase activity (BUChE) in plasma samples. Parasitemia were progressive increase and parasites were observed in the peripheral blood of all infected animals one day post-inoculation. AChE activity was not altered in cerebrum and cerebellum tissues. AChE activity in blood significantly decreased in the T3 and T5 groups (26.63 and 25.86 mU/l mol Hb) compared with the control (37.84 mU/l mol Hb). In addition BUChE activity in plasma was lower in the T3 (7.01 μmol BTC hydrolyzed/h/mL) than the T5 and C groups (9.84 and 12.00 μmol BTC hydrolyzed/h/mL). This study therefore, shows that reductions in the activity of cholinesterase occur in acute infection by T. evansi in rats and this demonstrates an important change occurring in animals infected by the protozoan and may indicate a potential role the enzymes play in the mechanism of disease.

This study aimed at evaluating biochemical changes of cats (Felis catus) experimentally infected with Trypanosoma evansi. Seven animals were infected with 10(8) blood trypomastigotes per animal and six were used as controls. Blood smears were performed daily for 56 days and the hepatic, renal and muscular parameters in blood serum were evaluated at days 0, 7, 21, 35 and 49. The protozoan was found in the bloodstream 24-48 h post-inoculation (PI) and irregular peaks of parasitemia were observed throughout the experiment. Muscular enzymatic activities (aspartate aminotransferase and creatine kinase) were increased in infected cats compared to controls. Increased concentrations of total proteins and globulins and decreased levels of albumin and albumin/globulin ratio were observed in infected group versus the controls values (P0.05). No alteration in serum activity of alanine aminotransferase, gamma-glutamyltransferase, creatinine and urea was observed in both groups.

Serum biochemical parameters and performance data were evaluated in broilers fed diets supplemented with antibiotics or essential oils from oregano, sage, rosemary, and pepper crude extract (OLES). Animals (n = 910) were distributed into five treatment groups, with seven replicates of 26 birds in each group: the control group (diet without additives); the group receiving an antibiotic growth promoter diet; and the groups T50, T100, and T150 (feed supplemented with 50, 100, and 150 mg/kg of OLES, respectively). After 42 days, 55 animals were randomly selected for serum biochemical profile analysis involving pancreatic, renal, and hepatic functions (lipase, amylase, urea, uric acid, aspartate aminotransferase, gamma glutamyltransferase, total cholesterol, high-density lipoprotein, triglycerides, total protein, albumin, globulins, and albumin/globulins ratio). Growth performance of broilers (body weight, weight gain, feed intake, alimentary conversion, and mortality) was also evaluated throughout the experiment period. The increase in serum levels of lipase, uric acid, urea, and aspartate aminotransferase suggests that OLES may cause kidney and liver impairment, mainly, at the higher dose. The OLES dose of 100 mg/kg is suitable for a final body weight and a weight gain similar to those observed in broilers supplemented with antibiotic growth promoters.

One of the main characteristics of the trypanosomosis is the development of anemia, although its pathogenesis still remains unclear. Therefore, the objective of this study was to discuss the possible pathogenesis of anemia in the infection by Trypanosoma evansi in cats. A study using an experimental model with T. evansi-infected cats reported changes in serum iron levels, alterations in the activity of enzymes of the cholinergic system, and oxidative stress correlated to normocytic, normochromic, and regenerative anemia. During this article, the aforementioned parameters will be simultaneously analyzed and related to hematological parameters that determine anemia in cats infected with this protozoan.

Changes in blood, plasma and brain cholinesterase activities in Trypanosoma evansi- infected cats were investigated. Seven animals were infected with 10 8 trypomastigote forms each and six were used as control. Animals were monitored for 56 days by examining daily blood smears. Blood samples were collected at days 28 and 56 post-inoculation to determine the activity of acetylcholinesterase (AChE) in blood and the activity of butyrylcholinesterase (BChE) in plasma. AChE was also evaluated in total brain. The activity of AChE in blood and brain, and the activity of BChE in plasma significantly reduced in the infected cats. Therefore, the infection by T. evansi influenced cholinesterases of felines indicating changes in the responses of the cholinergic system.

The aims of this study were to evaluate clotting disturbances in the acute infection by Trypanosoma evansi in rats and to investigate two possible causes. To address this issue, 21 2-month-old male rats were separated into three groups with seven animals each. The control group (group I) was composed of non-infected animals and group II was composed of 14 T. evansi-infected animals. This group was subdivided into two homogeneous groups. Animals from group II-a and from group II-b were euthanized at days 3 and 5 post-inoculation (PI), respectively. Hematological parameters were evaluated for monitoring of the disease. Infected animals showed anemia and leukocytosis 3 days PI and a significant decrease was observed in platelet count at days 3 and 5 PI. The prothrombin time (PT) and activated partial thromboplastin time (aPTT) were longer in the infected groups. The averages of the megakaryocyte count significantly increased (P

Five cats were experimentally inoculated with Trypanosoma evansi in order to evaluate the pathological changes induced by this protozoan infection. Clinical signs observed included vomiting, diarrhoea, hyperthermia, weight loss, facial oedema, corneal opacity, lymphadenopathy and hindlimb instability. Reduction in hematocrit was observed from 7 days post-infection (dpi) (P

This study aimed to assess the plasma lipid peroxidation and the susceptibility of erythrocytes to in vitro peroxidation as indicators of oxidative damage in erythrocytes and their roles in the pathogenesis of anemia during the early acute phase of Trypanosoma evansi infection in rats. Fifty male Wistar rats were randomly distributed into seven groups: three trypanosome-infected groups (T(2), T(4) and T(6); n=10 animals per group) and four uninfected controls (C(0), C(2), C(4) and C(6); n=5 animals per group). Animals from trypanosome-infected groups were inoculated intraperitoneally with 10(6) trypanosomes. Blood samples were collected by cardiac puncture before infection (day 0; group C(0)) or on the 2nd (C(2) and T(2)), 4th (C(4) and T(4)) and 6th (C(6) and T(6)) day post-infection (dpi). Samples were analyzed for red blood cell (RBC) count, hemoglobin (Hb) concentration, packed cell volume (PCV), plasma malondialdehyde (MDA) and in vitro peroxidation of erythrocytes. The mean values of the hematological indices gradually decreased in the infected rats compared with the control. MDA was significantly increased (P

The aim of this study was to investigate the efficacy of diminazene aceturate in the control of the infection by Trypanosoma evansi in cats. Fourteen animals were infected with 10(8) trypomastigote forms each and six were used as negative control (group A). Seven of the infected cats were used as positive control (group B) and seven were treated with diminazene aceturate (3.5 mg kg(-1)) for 5 consecutive days (group C). Biochemical and hematological parameters were evaluated during the experiment. Blood with anticoagulant was collected at day 49 post-inoculation and preserved in ethanol for DNA extraction. Samples were analyzed using PCR T. evansi-specific to assess the effectiveness of treatment. The treatment with diminazene aceturate had an efficacy of 85.7%. Alanine aminotransferase, gamma-glutamyltransferase, urea, and creatinine values remained within the normal physiological range in the treated cats. Hemogram was normalized in all the cured animals. Therefore, the therapy used is effective in controlling T. evansi in cats.

The objective of this study was to evaluate the lipid peroxidation and the susceptibility of erythrocytes to in vitro peroxidation as indicators of oxidative damage in erythrocytes and their roles in the pathogenesis of anemia during experimental Trypanosoma evansi infection in cats. Animals were divided into two groups: control and infected with T. evansi. Seven cats were infected with 108 trypomastigotes each, and parasitemia was estimated daily for 49 days by microscopic examination of smears. Hematological and biochemical parameters were evaluated for monitoring of the disease. Plasma lipid peroxidation (Thiobarbituric Acid Reactive Substances (TBARS)) and the susceptibility of erythrocytes to in vitro peroxidation were evaluated. Blood samples for analysis were collected at days 21 and 49 post-inoculation. TBARS level, indicated by MDA concentration, was higher in the infected group than in the control group in both analyzed periods, as well as the in vitro erythrocyte peroxidation (P

The aim of this study was to evaluate the concentrations of copper, iron and zinc in blood serum of cats experimentally infected with Trypanosoma evansi. Animals were divided into two groups: control and infected with T. evansi. The animals were infected with 10(8) trypomastigotes each and parasitemia was estimated daily for 56 days by microscopic examination of smears. Hematological and biochemical parameters were evaluated for monitoring of the disease. Serum metal levels were determined in blood samples collected at days 7, 28 and 56 of the experiment. Inductively coupled plasma optical emission spectrometry was used to measure the levels of copper, iron and zinc. Significant differences were observed among groups (P0.05). Increased levels of copper and decreased iron and zinc levels were observed. A decrease in the number of red blood cells was also observed 7 days after inoculation. Biochemical parameters were not altered. Therefore, the infection by T. evansi might alter the serum metal levels, causing metabolic disturbances in cats.

This study aimed at evaluating hemogram and erythropoietic changes in cats experimentally infected with Trypanosoma evansi. Thirteen adult female non-breeding Felix catus were separated into two groups: seven animals were infected with 10(8) trypomastigotes each, and six animals were used as negative controls. Animals were kept in air-conditioned rooms and blood smears were performed daily for 49 days. Blood samples were collected from the jugular vein at days 0, 7, 21, 35 and 49 and stored in blood-collecting tubes containing anticoagulant. Bone marrow was collected from the proximal epiphysis of the right femur at days 14 and 42 post-inoculation (PI). Total erythrocyte count, hematocrit and hemoglobin showed statistical differences among groups from the seventh day PI onwards (P0.05). The mean corpuscular volume and the mean corpuscular hemoglobin concentration remained normal, characterizing a normocytic-normochromic anemia. Reticulocyte count increased in the infected group from the 21st day onwards, but remained near normal values suggesting a mild regenerative anemia. Moreover, the myeloid:erythroid ratio significantly reduced at day 42 PI, evidencing a bone marrow hematopoietic response. Based on these results we conclude that cats infected with T. evansi have normocytic, normochromic, regenerative anemia.

Coagulopathy in cats (Felis catus) parasitized by Trypanosoma evansi in both the initial and chronic phases of the disease has been investigated. To address this issue, seven animals were infected with 108 trypomastigote forms each, and six were used as control. Animals were monitored for 56 days by examining daily blood smears. Blood samples for hematocrit, platelet counting, and fibrinogen levels were collect at 15-day intervals. Prothrombin time, activated partial thromboplastin time, and number of megakaryocytes were analyzed at days 14 and 42 post-inoculation. A decrease in hematocrit values and platelet counts and an increase in plasmatic fibrinogen concentration were observed in the infected cats (P

Biochemical and molecular research on parasites has increased considerably in trypanosomes in the recent years. Many of them have the purpose of identify areas, proteins and structures of the parasite which are vulnerable and could be used in therapy against the protozoan. Based on this hypothesis this study aimed to detect biochemically the enzyme adenosine deaminase (ADA) in Trypanosoma evansi, and to adapt an assay to the measurement of its activity in trypomastigotes. Firstly, the parasites were separated from the blood of mice experimentally infected with a DEAE-cellulose column. The ADA activity in trypomastigotes was evaluated at concentrations of 0.1, 0.2, 0.5, 0.6 and 0.8 mg of protein by spectrophotometry. ADA activity was observed in the parasites at all concentrations tested and its activity was proportional to the concentration of protein, ranging between 0.64 and 2.24 U/L in the lowest and highest concentration of protein, respectively. Therefore, it is possible to detect biochemically ADA in T. evansi, an enzyme that may be associated with vital functions of the parasite, similar to what occurs in mammals. This knowledge may be useful in the association of the chemotherapic treatment with specific inhibitors of the enzyme, in future studies.

The contamination of consumer food and animal feed with toxigenic fungi has resulted in economic losses worldwide in animal industries. Mycotoxins are highly biologically reactive secondary metabolites and can inhibit protein synthesis and cell multiplication. Considering the cytotoxicity of mycotoxins, this experiment was performed to determine the in vitro influence of ochratoxin A, deoxynivalenol and zearalenone on lipid peroxidation in lymphocytes of broiler chickens at different concentrations. This study has also evaluated whether the presence of these mycotoxins changes the acetylcholinesterase activity in lymphocytes, which is involved in the regulation of immune and inflammatory responses. Blood lymphocytes of broiler chickens were isolated through density gradient centrifugation and incubated with the respective mycotoxins at concentrations of 0.001, 0.01, 0.1 and 1 μg/mL. Lipid peroxidation, which was evaluated through the amount of malondialdehyde measured in a thiobarbituric acid-reactive species test, and the enzymatic activity were analyzed at 24, 48 and 72 h. Results of the lipid peroxidation evaluation showed an increasing cytotoxicity relation: ochratoxin A > deoxynivalenol > zearalenone. Conversely, cytotoxicity was valued as zearalenone > deoxynivalenol > ochratoxin A in relation to the acetylcholinesterase enzymatic activity. At a concentration of 1 μg/mL, ochratoxin A and deoxynivalenol induced the highest cellular oxidative stress levels and the highest enzymatic activity at the majority of time points. However, the same mycotoxins, except at 1 μg/mL concentration, induced a reduction of lymphocytic lipid peroxidation 72 h after incubation, suggesting the action of a compensatory mechanism in these cells.

SUMMARYThis study aimed to evaluatein vitroandin vivotrypanocidal activity of free and nanoencapsulated curcumin againstTrypanosoma evansi. In vitroefficacy of free curcumin (CURC) and curcumin-loaded in lipid-core nanocapsules (C-LNCs) was evaluated to verify their lethal effect onT. evansi. To perform thein vivotests,T. evansi-infected animals were treated with CURC (10 and 100 mg kg−1, intraperitoneally [i.p.]) and C-LNCs (10 mg kg−1, i.p.) during 6 days, with the results showing that these treatments significantly attenuated the parasitaemia. Infected untreated rats showed protein peroxidation and an increase of nitrites/nitrates, whereas animals treated with curcumin showed a reduction on these variables. As a result, the activity of antioxidant enzymes (superoxide dismutase and catalase) differs between groups (Pin vitroresulted in a better parasitaemia control, an antioxidant activity and a protective effect on liver and kidney functions ofT. evansi-infected adult male Wistar rats.

SUMMARYThe aim of this study was to assess the effect of sulfamethoxazole/trimethoprim (ST) supplemented with diphenyl diselenide and sodium selenite in experimental toxoplasmosis, on oxidant/antioxidant biomarkers and cytokine levels. Eighty-four BALB/c mice were divided in seven groups: group A (negative control), and groups B to G (infected). Blood and liver samples were collected on days 4 and 20 post infection (p.i.). Levels of thiobarbituric acid (TBA) reactive substances and advanced oxidation protein products (AOPP) were assessed in liver samples. Both biomarkers were significantly increased in infected groups on day 4 p.i., while they were reduced on day 20 p.i., compared with group A. Glutathione reductase (GR) activity significantly (Pγwas significantly increased (PPPP