A pilot study of chemotherapy combinations in rats: Focus on mammary cancer treatment in female dogs.

The use of combined chemotherapy is an essential alternative in treating breast cancer. However, knowledge of the pharmacokinetics of drugs is necessary to obtain maximum efficiency of the protocol and reduce adverse reactions. This study suggests for the first time the effect of the association of carboplatin with ivermectin and carboplatin with cyclophosphamide. This investigation was performed with 36 healthy Wistar rats, divided into four groups: group control, carboplatin (C), carboplatin preceded by ivermectin (C + IV), and carboplatin associated with cyclophosphamide (C + CI). Plasma concentrations quantification was performed using the High-Performance Liquid Chromatographic (HPLC) equipment with an Ultraviolet (UV) detector at eight different time points. Then, the animal was euthanized and necropsied. The bioanalytical method was validated for the two matrices (dogs and rats' plasma), with full validation in female dogs and partial validation in rats, as recommended by the EMA. In both matrices, the method was linear and reproducible. Here, we show the results in female rats' plasma. When comparing the experimental rats' groups (C; C + IV, and C + CI), there is a tendency to increase the bioavailability of carboplatin when used in association, a slight increase for C + IV and more evident to the C + CI group with an AUC rise higher than 2-fold (AUC0-∞ = 2983.61 for C; 4459.06 for C + CI; 7064.68 for C + CI min·mg·mL-1). The blood count, biochemistry profile, and histopathology of the organs revealed only alterations inherent to the metabolic effects of the drugs used. The carboplatin association with ivermectin appeared safe for this pilot group. We believe the carboplatin dose can be maintained without risk to the patient. However, in the carboplatin association with cyclophosphamide, a slight reduction in carboplatin's amount is suggested, seeking to avoid increased effects due to cyclophosphamide. Thus, studies with a more significant number per group must confirm the relevance of this pilot study.

Feline Leishmaniasis Caused by Leishmania infantum: Parasite Sequencing, Seropositivity, and Clinical Characterization in an Endemic Area From Brazil.

Zoonotic leishmaniasis caused by Leishmania infantum is a disease of One Health concern since human and animal cases and environmental damage are interconnected. L. infantum has a complex epidemiological cycle with multiple hosts, including mammals-humans, domestic, and wild animals-and arthropod vectors. Knowledge on mammal infections in endemic areas is crucial for developing control strategies. This work aimed to detect and characterize L. infantum infection in domestic cats from areas where human and canine leishmaniasis cases occur. No cases of feline leishmaniasis (FeL) had been previously reported in those areas. Five municipalities from Bahia state were chosen, comprising 2,480.8 km2 with 1,103,866 inhabitants. Ninety domiciliated and/or sheltered cats underwent clinical examination and serology by a rapid reference test recommended by the Brazilian government. Cytology, PCR, and parasite DNA sequencing were performed in bone marrow samples. Rapid tests detected antibodies in 5.6% (5/90) of the cats. Leishmania infantum infection was confirmed in 7.8% (7/90) of the cats by PCR, sequencing, and parasite isolation. Three out of the five municipalities (60%) had infected cats, and PCR positivity varied from 6.9 to 29%. One cat was categorized as harboring active L. infantum infection with amastigote forms in bone marrow smears. No clinical signs were detected at the first clinical exam, but 1 month later the cat developed severe FeL. The cat isolate was grown in culture, typed and its DNA sequence was homologous to the L. infantum reference strain (PP75). In conclusion, cats are potential hosts and may acquire L. infantum in endemic areas where canine and human cases occur. For cats, the need for surveillance, differential diagnosis and clinical care is highly recommended since a fast clinical progression of FeL developed in a subclinical animal. An accurate standardized immunodiagnostic assay for FeL is warranted.

Histopathological Parameters of Canine Bone Marrow in Cell-Block Preparations.

Studies using the cell-block technique for bone marrow (BM) analysis are limited in the veterinary literature. This work assessed whether the histopathological analysis of canine BM was feasible using cell-block cytoinclusions prepared from fine-needle sternal aspirate samples. Eight clinically healthy young-to-middle-aged dogs underwent both fine-needle sternal aspiration for BM cell-block (BM-Cb) processing and iliac-crest BM core biopsy (BM-B). Histopathologic parameters were compared between the 2 methods. There were no statistically significant histopathological differences between hematopoietic tissue areas (P = .6294) in the BM-Cb and BM-B sections, and they had similar microscopic characteristics and microarchitecture. Cellularity and reticulin-fiber staining were equivalent in the BM-Cb and BM-B preparations in 87.5% (7/8) and 100% (8/8) of the sections, respectively. However, the quantitative results of the megakaryocytic series differed between BM-Cb and BM-B in 37.5% (3/8) of the sections, and the myeloid:erythroid (M:E) ratios differed between the 2 methods in 25% (2/8). These preliminary data indicate that cell-block preparations made from sternal fine-needle aspiration samples warrant continued evaluation in a larger number of animals, including those with various diseases affecting the bone marrow.

Treatment with isolated gold nanoparticles reverses brain damage caused by obesity.

In this study, we performed two experiments. In the first experiment, the objective was to link gold nanoparticles (GNPs) with sodium diclofenac and/or soy lecithin and to determine their concentration in tissues and their toxicity using hepatic and renal analyzes in mice to evaluate their safety as therapeutic agents in the subsequent treatment of obesity. In the second experiment, we evaluated the effect of GNPs on inflammatory and biochemical parameters in obese mice. In the first experiment, we synthesized and characterized 18 nm GNPs that were administered intraperitoneally in isolation or in association with sodium diclofenac and/or soy lecithin in mice once daily for 1 or 14 days. Twenty-four hours after the single or final administration, the animals were euthanized, following which the tissues were removed for evaluating the concentration of GNPs, and serum samples were collected for hepatic and renal analysis. Hepatic damage was evaluated based on the levels of alanine aminotransferase (ALT), whereas renal damage was evaluated based on creatinine levels. A higher concentration of GNPs was detected in the tissues upon administration for 14 days, and there were no signs of hepatic or renal damage. In the second experiment, the mice were used as animal models of obesity and were fed a high-fat diet (obese group) and control diet (control group). After eight weeks of high-fat diet administration, the mice were treated with saline or with GNPs (average size of 18 nm) at a concentration of 70 mg/L (70 mg/kg) once a day, for 14 days, for 10 weeks. Body weight and food intake were measured frequently. After the experiment ended, the animals were euthanized, serum samples were collected for glucose and lipid profile analysis, the mesenteric fat content was weighed, and the brains were removed for inflammatory and biochemical analysis. In obese mice, although GNP administration did not reduce body and mesenteric fat weight, it reduced food intake. The glucose levels were reversed upon administration of GNPs, whereas the lipid profile was not altered in any of the groups. GNPs exerted a beneficial effect on inflammation and oxidative stress parameters, without reverting mitochondrial dysfunction. Our results indicate that the intraperitoneal administration of GNPs for 14 days results in a significant GNP concentration in adipose tissues, which could be an interesting finding for the treatment of inflammation associated with obesity. Based on the efficacy of GNPs in reducing dietary intake, inflammation, and oxidative stress, they can be considered potential alternative agents for the treatment of obesity.

Quality control in veterinary blood banks: evaluation of canine platelet concentrates stored for five days.

BACKGROUND: Platelets undergo structural, biochemical and functional alterations when stored, and platelet storage lesions reduce platelet function and half-life after transfusion. The objective of this study was to evaluate stored canine platelet concentrates with platelet aggregation, flow cytometry and biochemistry assays. Twenty-two bags of canine platelet concentrates were obtained by the platelet-rich plasma method and were assessed on days 1, 3 and 5 after collection. Parameters such as platelet counts, residual leukocytes, platelet swirling, glucose, lactate, pH, CD62P expression (platelet activation), JC-1 (mitochondrial function) and annexin V (apoptosis and cell death) were assessed. RESULTS: Over the five days of storage there was a significant decrease in glucose, HCO3, pCO2, ATP, pH, swirling and mitochondrial function, associated with a significant increase in lactate levels and pO2. At the end of storage pH was 5.9 ± 0.6 and lactate levels were 2.8 ± 1.2 mmol/L. Results of the quality parameters evaluated were similar to those reported in human platelets studies. The deleterious effects of storage were more pronounced in bags with higher platelet counts (> 7.49 × 1010/unit), suggesting that canine platelet concentrates should not contain an excessive number of platelets. CONCLUSIONS: Quality parameters of canine platelets under standard storage conditions were similar to those observed in human platelets. Our results have potential to be used for the routine evaluation and quality control in veterinary blood banks.

Comparison of JC-1 and MitoTracker probes for mitochondrial viability assessment in stored canine platelet concentrates: A flow cytometry study.

Mitochondria perform crucial roles in many biochemical processes, and mitochondrial depolarization is an early sign of platelet apoptosis. The mitochondrial membrane potential is usually evaluated through JC-1 probe, but it can also be assessed with MitoTracker probes. Our aim was to evaluate mitochondrial viability in stored canine platelet concentrates (PCs) with the fluorescent probes JC-1 and MitoTracker. Platelets from 22 canine PCs were stained with JC-1 and MitoTracker probes on days 1, 3, and 5 of storage. Data on metabolic parameters were also collected for correlation studies. Results of JC-1 and MitoTracker revealed a decrease in mitochondrial membrane potential in day 5 of storage compared to days 1 and 3, providing evidence of mitochondrial depolarization, a finding that was confirmed by the data on metabolic parameters. MitoTracker probes also added information regarding platelet swelling. In conclusion, MitoTracker probes offered a more complete mitochondrial analysis in the evaluation of stored canine PCs. © 2018 International Society for Advancement of Cytometry.

Effects of four additive solutions on canine leukoreduced red cell concentrate quality during storage.

BACKGROUND: Additive solutions (AS) and prestorage leukoreduction (LR) are important tools used to maintain erythrocyte viability during storage and avoid transfusion reactions in recipients, respectively. OBJECTIVES: The purpose of the study was to determine the efficacy of a WBC filter (Immugard IIIRC) and compare the effect of 4 AS (phosphate-adenine-glucose-guanosine-gluconate-mannitol [PAGGGM], saline-adenine-glucose-mannitol [SAGM], Adsol, Optisol) on the in vitro quality of canine leukoreduced packed RBC units (pRBC) stored for 41 days. METHODS: Five hundred milliliters of blood were collected from 8 healthy dogs each into 70 mL of citrate-phosphate-dextrose (CPD) solution, and were leukoreduced by a polyurethane filter. pRBC of each dog were divided equally into 4 bags containing a different AS. Bags were stored for 41 days at 4°C and evaluated every 10 days. Variables analyzed included pH, PCV, and% hemolysis, and lactate, glucose, potassium, sodium, ATP, and 2,3-diphosphoglycerate (2,3-DPG) concentrations. RESULTS: The LR resulted in residual WBC counts comparable to human standards. During storage, pH, and glucose, 2,3-DPG, and ATP concentrations decreased, and hemolysis, and lactate, sodium, and potassium concentrations increased (P < .05). Significant differences between AS were seen in the glucose and sodium concentrations, due to the composition of AS. Also, the pH maintained by PAGGGM at day 21 was significantly higher than that seen with SAGM or Adsol. CONCLUSIONS: All AS used gave satisfactory results during the first 21 days of storage based on the degree of hemolysis, and on ATP and 2,3-DPG concentrations. When compared with day 1 values, significant changes were seen in these variables by day 31 with all AS.

Epidemiological, clinical and immunohistochemical aspects of canine lymphoma in the region of Porto Alegre, Brazil / Aspectos epidemiológicos, clínicos e imuno-histoquímicos do linfoma canino na região de Porto Alegre

This paper describes the epidemiological, clinical and immunohistochemical characteristics of canine lymphomas diagnosed in the region of Porto Alegre, Brazil. Thirty dogs were enrolled in the study; most of them were male (60 percent), mixed-breed (23 percent) and middle-aged or older. The majority (87 percent) of affected dogs showed the multicentric form. The B-cell phenotype was most frequently detected (62 percent); 37 percent of the animals were in clinical stage IV, and 83 percent were classified as sub-stage "b". Lymphadenopathy was observed in 67 percent of the cases, and dyspnea, prostration, decreased appetite and vomiting were the most common clinical signs encountered. Anemia was a frequently encountered laboratory alteration (57 percent), as were leukocytosis (40 percent), thrombocytopenia (33 percent), lymphopenia (30 percent), hyperglobulinemia (20 percent) and hypercalcemia (13 percent). The results of this study indicate that the clinical features of dogs with lymphoma in the region of Porto Alegre are similar to those observed worldwide.

Esse trabalho apresenta os achados epidemiológicos, clínicos e imuno-histoquímicos de linfomas caninos diagnosticados na região de Porto Alegre. Trinta cães foram incluídos no estudo; sendo principalmente machos (60 por cento), sem raça definida (23 por cento) e de meia-idade a idosos. A maioria (87 por cento) dos cães afetados apresentou a forma anatômica multicêntrica. O imunofenótipo B foi detectado com mais frequência (62 por cento); 37 por cento dos cães apresentavam estadiamento clínico IV e 83 por cento encontravam-se no subestádio "b". Linfadenopatia foi observada em 67 por cento dos casos; outros sinais clínicos comumente detectados foram dispneia, prostração, diminuição do apetite e vômitos. Anemia foi a alteração laboratorial mais frequente (57 por cento), seguida por leucocitose (40 por cento), trombocitopenia (33 por cento), linfopenia (30 por cento), hiperglobulinemia (20 por cento), hiperproteinemia (17 por cento) e hipercalcemia (13 por cento). Os resultados do presente estudo indicam que as características epidemiológicas e clínicas de cães com linfoma na região de Porto Alegre são semelhantes às observadas em todo o mundo.

Clinicopathologic and electrocardiographic features of Akita dogs with high and low erythrocyte potassium phenotypes.

BACKGROUND: Canine RBCs are expected to have high sodium and low potassium (LK) concentrations. However, some dogs have a low sodium and high potassium (HK) phenotype due to a Na-K pump in the mature RBC membrane. Awareness of this particularity avoids misinterpretation of hyperkalemia in affected dogs. OBJECTIVE: The purpose of this study was to compare the CBCs, serum biochemistry profiles, urinalysis data, and electrocardiography data in HK and LK Akita dogs. METHODS: In this study, the intra-RBC potassium (KRBC) was measured in 48 healthy adult Akita dogs. The HK group included dogs with KRBC concentration at least 5-fold that of plasma, while the LK dogs served as controls. RESULTS: The HK phenotype was determined in 10 dogs (21%). With the exception of higher plasma potassium concentrations in the HK group compared with LK controls (6.6 mmol/L vs 4.4 mmol/L, P < .001), there were no other differences in serum biochemistry variables. In hematology, mean HGB concentration (13.5 vs 15.9, P < .001), PCV (42.9 vs 46.7, P = .009), RBC (6.1 vs 7.4, P < .001), and MCHC (31.3 vs 34.0, P < .001) were significantly lower in the HK group, while MCV (70.0 vs 63.4, P < .001) was higher compared with the LK controls. There were no significant differences in urinalysis or electrocardiography data between groups. CONCLUSIONS: HK Akita dogs showed significant differences in several hematologic variables, as well as higher plasma potassium concentration. Therefore, the HK phenotype should be considered in the interpretation of these variables in this breed.

Parâmetros hematológicos do roedor fossorial Ctenomys lami (Rodentia, Ctenomidae) no estado do Rio Grande do Sul / Hematological parameters of fossorial rodent Ctenomys lami (Rodentia, Ctenomidae) in Rio Grande do Sul, Brazil

O perfil hematológico sanguíneo foi determinado em três populações de Ctenomys lami, em áreas denominadas A e B, impactadas pela bovinocultura, e C, sem impacto antrópico, ambas no sul do Brasil. Sessenta e dois animais foram coletados ao total. Os valores de hematócrito (Ht), hemoglobina (Hb) e eritrócitos apresentaram diferenças significativas entre machos e fêmeas. Os valores médios de Ht e a Hb encontrados na espécie foram mais baixos em comparação com os de outras espécies de roedores subterrâneos, podendo estes valores estarem relacionados ao habitat de forrageio ou às características do solo. Também foram encontradas diferenças significativas nas médias de hemoglobina, CHCM e linfócitos em animais das áreas A e B em relação à área C. O valor da média dos hematócritos dos animais entre as áreas foi mais elevado nas áreas A e B, porém significativamente diferentes entre A e C. Algumas dessas alterações sugerem a relação dos valores encontrados com o estresse dos animais em relação a áreas impactadas. Variações significativas no VCM foram encontradas entre os animais das áreas A e C, e também nas plaquetas destes entre as áreas A e B. Não foram observados Corpúsculo de Kurloff no sangue dos animais analisados. Os valores hematológicos encontrados nesses espécimes de Ctenomys lami fornecem informações importantes sobre a espécie e podem ser úteis em outras pesquisas.

The hematological profile was determined in three populations of Ctenomys lami that inhabits three different areas nominated as A and B, affected by cattle production, and C, without human impact, all of them in southern Brazil, under the same geologic formation. Sixty two individuals were collected. The packed cell volume (PCV) values, hemoglobin (Hb) and red blood cell (RBC) count presents statistic significant differences between males and females. PCV and Hb values were lower in comparison with other subterranean rodents, which could be related to the food searching behavior or soil characteristic of the species. Significant statistic differences were found to for Hb, mean corpuscular hemoglobin concentration (MCHC) and lymphocytes between animals belonging to A and C areas. The PCV values were higher for animals from areas A and B, with significance only between A and C. Some of these alterations may suggest a modification on stress levels of the animals inhabiting areas A and B with anthropic alterations, or maybe just an inherited characteristic. Significant statistic differences were found either in mean corpuscular volume (MCV) between A and C areas, and platelets between A and B areas. No Kurloff cells were observed in the blood smear from analyzed individuals. The hematological values founded for Ctenomys lami are an important information for this species, which can be used for future research and management.

Parâmetros de bioquímica sérica de machos, fêmeas e filhotes de Araras canindé (Ara ararauna) saudáveis mantidas em cativeiro comercial / Serum biochemical parameters of healthy male, female and young blue-and-yellow macaws (Ara ararauna) bred in captivity

O Brasil possui um considerável número de espécies de psitacídeos catalogados, perfazendo cerca de 80 espécies, sendo que as Araras canindé (Ara ararauna), uma das maiores representantes dessa ordem, podem ser encontradas em florestas nas diversas regiões brasileiras. O Instituto Brasileiro do Meio Ambiente e dos Recursos Naturais Renováveis (IBAMA) normalizou a comercialização de animais da fauna silvestre provenientes de criadouros e conseqüentemente ocorreu um aumento do número destes animais como de estimação. Atualmente, há pouco conhecimento sobre os parâmetros clínicos e laboratoriais de espécies silvestres em cativeiro. O presente trabalho teve como objetivo determinar parâmetros de bioquímica sangüínea de Araras canindé (Ara ararauna) saudáveis de sexo e faixa etária distintas mantidas em um criatório comercial com alimentação e manejo controlados e padronizados. Foram colhidas amostras de sangue de 35 araras canindé (11 filhotes e 24 adultos) e remetidas ao Laboratório de Análises Clínicas Veterinárias da Universidade de Passo Fundo (UPF) para determinação dos seguintes indicadores bioquímicos: ácido úrico (AU), albumina (ALB), aspartato aminotransferase (AST), cálcio (Ca), colesterol (Col), creatina quinase (CK), fosfatase alcalina (FA), fósforo inorgânico (Pi), frutosamina (Fru), gama glutamil transferase (GGT), proteínas totais (PT) e uréia (UR). Entre as aves adultas, foram encontradas diferenças significativas nos valores de CK (superior nos machos), Ca e AU (superiores nas fêmeas). Entre aves adultas e filhotes foram constatadas diferenças significativas nos valores de AU, Ca, ALB, COL, FA, Pi e FRU. A maioria das alterações detectadas pode ser relacionada à diferença na dieta fornecida e à condição fisiológica de cada categoria de aves. Os dados obtidos podem ser utilizados como parâmetros de referência para as araras canindé brasileiras.

Several psittacines have been cataloged in Brazil, totaling nearly 80 species. The blue-and-yellow macaw (Ara ararauna), the most representative species of the Psittaciformes order, lives in the forest areas of different Brazilian regions. IBAMA, Brazilian environmental protection agency, has authorized the trade of wild animals raised in breeding facilities; consequently, their use as household pets has increased ever since. Currently, too little is known about clinical and laboratory parameters of wild species bred in captivity. The aim of the present study was to determine the serum biochemical parameters of healthy blue-and-yellow macaws (Ara ararauna) of different sexes and ages reared in a commercial breeding facility under controlled and standardized feeding and management practices. Blood samples were collected from 35 blue-and-yellow macaws (11 fledglings and 24 adults) and sent to the Laboratory of Veterinary Clinical Pathology of Universidade de Passo Fundo (UPF) for measurement of the following biochemical parameters: uric acid (UA), albumin (Alb), aspartate aminotransferase (AST), calcium (Ca), cholesterol (Chol), creatine kinase (CK), alkaline phosphatase (AP), inorganic phosphorus (iP), fructosamine (Fru), gamma-glutamyl transferase (GGT), total protein (TP) and urea nitrogen (Ur). There were significant differences among adult macaws with regard to CK levels (higher in males) and to Ca and UA levels (higher in females). Differences were noted among adults and fledglings with respect to UA, Ca, Alb, Chol, AP, iP and Fru. Most of the findings can be ascribed to the different feeding regimens and ages of the birds analyzed. The data obtained herein can be used as reference parameters for Brazilian blue-and-yellow macaws.