Hematological Normality, Serum Biochemistry, and Acute Phase Proteins in Healthy Beef Calves in the Brazilian Savannah.

The Brazilian savannah region, characterized by high average temperatures, well-defined rainy and dry seasons, soil with low productive potential, and high pressure for parasitic diseases, is home to the highest percentage of the beef herd, which is the world's largest commercial beef producer. Therefore, breeds that present rusticity combined with productivity are the focus of research in cattle breeding in the region. Considering their geographic particularities and their effects on the animals' blood parameters, the objective was to study the behavior of hematological variables, serum biochemistry, and acute phase proteins in beef calves at weaning age. Jugular blood samples were collected in a single day from 30 weaning calves (about eight months old and 200 kg of body weight) from the following breeds: Nellore, Senepol, Nellore × Aberdeen Angus cross, Nellore × Senepol cross, and Nellore × Aberdeen Angus × Senepol cross. Hematological data were obtained using an automatic cell counter, serum biochemical measurements were obtained using commercial kits, and the electrophoretogram was obtained using the SDS-page technique. In general, the results were consistent with data already published in similar situations regarding health status, age, and level of metabolic activity. However, differences observed between groups can be explained by differences observed in other concurrent variables like temperament. The pure zebu breed was more reactive than pure taurine. While crossbreds showed intermediate values, and parasitic infestation, the pure taurine breed with higher parasite infestation, while the zebu breed had lower values, which produced effects in some laboratory tests, and generated differences between breeds. In conclusion, the reference intervals available for healthy animals can be routinely used without interference from the geographic region for animals produced without nutritional failures as long as changes are recorded in pathological, infectious, metabolic, or nutritional deficiency situations. However, it is suggested that a study covering a larger number of herds may demonstrate a greater geographic effect on the studied variables.

Renal Abnormalities Caused by Canine Distemper Virus Infection in Terminal Patients.

The aim of this study was to analyze the glomerular and tubular alterations in dogs with terminal distemper through light microscopy, immunofluorescence, and electron microscopy. Thirteen animals with a molecular diagnosis of distemper and neurological signs were selected. As a control group, 10 clinically healthy animals with no manifestations or signs of disease and with negative tests for Ehrlichia sp., Anaplasma sp., and Babesia sp. were included in this study. Renal tissue was evaluated by light microscopy, topochemistry for DNA/chromatin, and video image analysis to detect the nuclear phenotypes of the renal tubular epithelial cells (RTECs), immunofluorescence, and transmission electron microscopy. Results showed that dogs with distemper exhibited anemia, hypergammaglobulinemia, and proteinuria. Creatinine in the distemper group was lower compared to the control group (p = 0.0026), but there was no significant difference in relation to urea (p = 0.9876). Although this alteration may be due to the smaller muscle mass observed in animals with distemper, it probably is not of clinical importance. Glomerular and tubular lesions were confirmed by light microscopy in 84.6% of these animals. Additional findings in the animals with distemper included deposition of different classes of immunoglobulins, particularly IgM in 92.3% of the cases, fibrinogen deposition in 69.2% of the cases as assessed by immunofluorescence, alterations in the nuclear phenotypes of the RTEC characterized by condensation of chromatin, loss of DNA and reduction in the nuclear shape, and the presence of subendothelial and mesangial electron-dense deposits. These findings confirm the existence of renal alterations related to terminal distemper.

Liquid biopsy can detect BRCA2 gene variants in female dogs with mammary neoplasia.

Mammary tumours (MT) are one of the most prevalent malignancies in female dogs and women. Currently, molecular analyzes have shown that each tumour type presents its own genetic signature. In this context, liquid biopsy allows a comprehensive genetic characterisation of the tumour, enabling early diagnosis and personalised treatment of patients. In women, deleterious mutations inherited in BRCA2 gene are associated with an increased risk of breast cancer, resistance to therapies and worse prognosis. In female dogs, there are many divergent data on the involvement of BRCA2 gene with mammary carcinogenesis and what its pathogenic potential is. Therefore, the objective was to identify BRCA2 gene variants in 20 plasma DNA samples, from 10 newly diagnosed dogs with mammary cancer (RD), five control (CTR) and five mastectomized patients. Eleven single nucleotide polymorphisms (SNPs) were detected, most of them in the exon 11 and two indels (deletion/insertion) in the BRCA2 gene. However, there was no statistically significant difference in the SNPs/indels detected between the groups. In addition, only one SNP (p.T1425P) and one deletion (p.L2307del) were considered deleterious using in silico computational models. Interestingly, most common variants were present in the plasma of all groups, except for the Ile2614Thr, Ile2614Val, Thr1425Pro and p.L2307del variants. Thus, we observed that SNPs are common in the BRCA2 gene of female dogs with MT, with a similar condition identified in women with breast cancer. Liquid biopsy approach in dogs with MT is useful for genetic and therapeutic proposals.

Renal Magnesium Handling: A Comparison Between Dogs With Chronic Kidney Disease and Healthy Elderly Dogs.

Chronic kidney disease (CKD) and aging are known to possibly cause a progressive reduction in glomerular filtration rate, which may be associated with an increase in fractional excretion of electrolytes due to an adaptive response of the remaining functioning nephrons. However, the behavior of magnesium excretion has not been studied in CKD and healthy elderly dogs. The objective was to evaluate the fractional excretion of magnesium (FEMg) in dogs with (CKD) compared to healthy elderly dogs. Sixteen healthy elderly dogs and 43 dogs with CKD were divided into 3 groups (CKD 2 [n = 14], CKD 3 [n = 17], CKD 4 [n = 12]), in accordance with the current International Renal Interest Society (IRIS) criteria that were used in this study. Blood samples were obtained by jugular venipuncture and urine samples were obtained by cystocentesis. The FEMg was evaluated at a single time point in both urine and blood samples. FEMg was significantly higher in dogs with CKD compared to healthy elderly dogs, especially in advanced stages. This preliminary study demonstrates that FEMg may be altered in dogs with CKD. Further research is warranted to elucidate magnesium's potential role in cardiovascular and arterial calcification in dogs with CKD as observed in humans with CKD.

Glomerulotubular pathology in dogs with subclinical ehrlichiosis.

Subclinical stage of ehrlichiosis is characterized by absence of clinical or laboratory alterations; however, it could lead to silent glomerular/tubular changes and contribute significantly to renal failure in humans and animals. The aim of this study was to evaluate glomerular and tubular alterations in dogs with subclinical ehrlichiosis. We evaluated renal biopsies of 14 bitches with subclinical ehrlichiosis and 11 control dogs. Samples were obtained from the left kidney, and the tissue obtained was divided for light microscopy, immunofluorescence, and transmission electron microscopy. Abnormalities were identified by light microscopy in 92.9% of dogs with ehrlichiosis, but not in any of the dogs of the control group. Mesangial cell proliferation and synechiae (46.1%) were the most common findings, but focal segmental glomerulosclerosis and ischemic glomeruli (38.4%), focal glomerular mesangial matrix expansion (30.7%), mild to moderate interstitial fibrosis and tubular atrophy (23%), and glomerular basement membrane spikes (23%) were also frequent in dogs with ehrlichiosis. All animals with ehrlichiosis exhibited positive immunofluorescence staining for immunoglobulins. Transmission electron microscopy from dogs with ehrlichiosis revealed slight changes such as sparse surface projections and basement membrane double contour. The subclinical phase of ehrlichiosis poses a higher risk of development of kidney damage due to the deposition of immune complexes.

Toxicity of a methotrexate metronomic schedule in Wistar rats.

Metronomic chemotherapy is a relevant strategy that uses low doses of antineoplastic drugs for sustained periods to control tumor growth, an alternative frequently utilized in veterinary patients. This work aimed to evaluate the toxic effects of a metronomic oral dose of methotrexate (MTX) for 45 days in tumor-free Wistar rats when compared with control animals. Clinical alterations, body weight, food, and water intake were monitored daily, and bone marrow suppression, hematological, biochemical, and histopathological analyses were performed at three points (days 30, 45, and 60). MTX-treated animals did not demonstrate severe systemic involvement. At 30 days, compared with control animals, MTX-treated animals showed significant leukocytosis (11.9 ± 2.3 vs. 7.8 ± 0.2 106/µL; P < .05) and augmentation of immature myeloid populations from bone marrow (9.0 ± 0.8 vs. 6.5 ± 1.5%; P < .05), and at 60 days, treated animals showed significant neutrophilia (35.0 ± 11.0 vs. 23.00 ± 3.0%; P < .05), depletion of bone marrow lymphocytes (8.2 ± 0.7 vs. 11.5 ± 1.9%; P < .05), and immature myeloid populations (7.2 ± 0.7 vs. 8.3 ± 0.6%; P < .05). At a histopathological level, splenic hypoplasia and respiratory inflammatory lesions were significant when compared with control animals, presenting mild to moderate myelotoxicity, immune suppression, and associated clinical compromise that persisted beyond treatment withdrawal. This suggested that MTX metronomic toxicity should not be neglected owing to the observed residual side-effects and special care should be taken regarding myelosuppression.

Hepatozoon sp. gamonts as an accidental finding in synovial liquid from an injured maned wolf (Chrysocyon brachyurus) in southeastern Brazil / Gamontes de Hepatozoon sp. como achado acidental em líquido sinovial de lobo guará (Chrysocyon brachyurus) no sudeste do Brasil

Abstract A free-living, adult male maned wolf (Chrysocyon brachyurus) was referred to the Governador "Laudo Natel" - FCAV/Unesp veterinary hospital after being found with skin lesions and a fracture on the right pelvic limb, which had to be amputated due to compromised integrity. Around 20 days later, bilateral accentuated swollen on humerus-radius-ulna articulation was observed. The synovial liquid was drained and sent to the laboratory for synovial cytology with Rosenfeld staining that revealed predominantly degenerated neutrophils with karyolytic chromatin associated with intracellular inclusions suggestive of Hepatozoon sp. gametocytes. Blood and synovial liquid samples were submitted to molecular analysis, aiming to amplify the Hepatozoon spp. 18S rRNA gene fragment. Despite the positioning of the found Hepatozoon sequence together with Hepatozoon canis previously detected in domestic carnivores, the BLAST analysis showed only 98% identity with H. canis. To the best of the authors' knowledge, this is the first time a Hepatozoon was detected in the synovial liquid by clinical pathology and molecular analyses.

Resumo Um lobo guará (Chrysocyon brachyurus) adulto, macho, de vida livre foi encaminhado para atendimento no hospital veterinário Governador "Laudo Natel" - FCAV/Unesp após ser encontrado com lesões de pele e fratura em membro pélvico direito, sendo amputado devido a comprometimento da integridade do membro. Aproximadamente 20 dias após a chegada ao hospital, foi notado acentuado aumento de volume bilateral em região de articulação úmero-rádio-ulnar. O líquido sinovial foi drenado e enviado para análise citológica com coloração de Rosenfeld, revelando a presença de neutrófilos degenerados com cromatina cariolítica associados a inclusões intracelulares sugestivas de gametócitos de Hepatozoon sp. Amostras de sangue e líquido sinovial foram submetidas a análises moleculares visando amplificar um fragmento do gene 18S rRNA de Hepatozoon spp. Apesar da sequência de Hepatozoon detectada se posicionar filogeneticamente no mesmo clado que H. canis previamente detectado em carnívoros domésticos, o resultado da análise do BLAST mostrou somente 98% de identidade com H. canis. De acordo com o conhecimento dos autores, esta é a primeira vez que Hepatozoon foi detectado no líquido sinovial por meio de patologia clínica e análises moleculares.

Hepatozoon sp. gamonts as an accidental finding in synovial liquid from an injured maned wolf (Chrysocyon brachyurus) in southeastern Brazil.

A free-living, adult male maned wolf (Chrysocyon brachyurus) was referred to the Governador "Laudo Natel" - FCAV/Unesp veterinary hospital after being found with skin lesions and a fracture on the right pelvic limb, which had to be amputated due to compromised integrity. Around 20 days later, bilateral accentuated swollen on humerus-radius-ulna articulation was observed. The synovial liquid was drained and sent to the laboratory for synovial cytology with Rosenfeld staining that revealed predominantly degenerated neutrophils with karyolytic chromatin associated with intracellular inclusions suggestive of Hepatozoon sp. gametocytes. Blood and synovial liquid samples were submitted to molecular analysis, aiming to amplify the Hepatozoon spp. 18S rRNA gene fragment. Despite the positioning of the found Hepatozoon sequence together with Hepatozoon canis previously detected in domestic carnivores, the BLAST analysis showed only 98% identity with H. canis. To the best of the authors' knowledge, this is the first time a Hepatozoon was detected in the synovial liquid by clinical pathology and molecular analyses.

Relação do proteinograma sérico e as fases folicular e luteal do ciclo estral em éguas / Relation of the serum proteins and the follicular and luteal phasesof the estrous cycle in mares

O objetivo deste estudo foi obter o perfil eletroforético das proteínas séricas em éguas cíclicas e verificar as diferenças entre as fases folicular e luteal do ciclo estral nesta espécie. Foram utilizadas 18 éguas, totalizando 36 amostras de soro, sendo duas de cada égua. As amostras foram colhidas no estro e no diestro. As proteínas séricas totais foram obtidas pelo método do Biureto, a partir da utilização de Kits comerciais (LABTEST®) e, as diferentes subfrações proteicas, por eletroforese em gel de poliacrilamida (SDS-PAGE). O eletroforetograma das proteínas séricas colocou em evidência a presença de 17 a 25 frações proteicas, cujos pesos moleculares variaram de 22 a 254 kDa. Identificaram-se duas proteínas ainda não nomeadas oficialmente, de massas moleculares (MM) 23 kDa e 144 kDa. Os valores médios ± SEM obtidos para cada variável no estro e no diestro, respectivamente, foram: proteínas totais (g/dL) 7,11 ± 0,07 e 7,36 ± 0,07; albumina (mg/dL) 4790,83 ± 69,10 e 5027,19 ± 69,10; α1 glicoproteína ácida (mg/dL) 4,90 ± 0,31 e 4,93 ± 0,31; ceruloplasmina (mg/dL) 15,28 ± 1,31 e 10,65 ± 1,31; haptoglobina (mg/dL) 22,70 ± 1,16 e 27,06 ± 1,16; transferrina (mg/dL) 329,00 ± 9,78 e 350,16 ± 9,78; IgA (mg/dL) 119,91 ± 6,30 e 107,03 ± 6,30; IgG (mg/dL) 1525,07 ± 40,18 e 1517,25 ± 40,18; MM 23 (mg/dL) 204,44 ± 8,61 e 219,79 ± 8,61; MM 144 (mg/dL) 22,13 ± 0,55 e 21,49 ± 0,55. Não houve diferença significativa das proteínas totais e suas frações do estro para o diestro. Conclui-se que as modificações hormonais durante as fases do ciclo estral da égua não interferem no proteinograma sérico.

This study aimed to obtain the electrophoretic profile of serum proteins in cyclic mares and to verify the differences between the follicular and luteal phases of the estrous cycle in this species. Eighteen mares were used, totaling 36 serum samples, two of each mare. Samples were collected both in estrus and in diestrus. Total serum proteins were obtained by the Biureto method, by using commercial kits (LABTEST®), while the different protein subfractions by polyacrylamide gel electrophoresis (SDS-PAGE). The electroforetogram of serum proteins evidenced the presence of 17 to 25 protein fractions, whose molecular weights ranged from 22 to 254 kDa. Two proteins that were not yet officially named were identified, of molecular weights (MW) of 23 kDa and 144 kDa. The mean values (± SEM) obtained for each variable in estrus and diestrus were, respectively: total proteins (g/dL) 7.11 ± 0.07 and 7.36 ± 0.07; albumin (mg/dL) 4790.83 ± 69.10 and 5027.19 ± 69.10; α1 acid glycoprotein (mg/dL) 4.90 ± 0.31 and 4.93 ± 0.31; ceruloplasmine (mg/dL) 15.28 ± 1.31 and 10.65 ± 1.31; haptoglobine (mg/dL) 22.70 ± 1.16 and 27.06 ± 1.16; transferrin (mg/ dL) 329.00 ± 9.78 and 350.16 ± 9.78; IgA (mg/dL) 119.91 ± 6.30 and 107.03 ± 6.30; IgG (mg/dL) 1525.07 ± 40.18 and 1517.25 ± 40.18; MW 23 (mg/dL) 204.44 ± 8.61 and 219.79 ± 8.61; MW 144 (mg/dL) 22.13 ± 0.55 and 21.49 ± 0.55. No significant difference was verified in total proteins and its fractions in estrus and diestrus. The hormonal changes during the specific stages of the estrous cycle of the mare do not interfere with the serum proteinogram.

Relação do proteinograma sérico e as fases folicular e luteal do ciclo estral em éguas / Relation of the serum proteins and the follicular and luteal phases of the estrous cycle in mares

O objetivo deste estudo foi obter o perfil eletroforético das proteínas séricas em éguas cíclicas e verificar as diferenças entre as fases folicular e luteal do ciclo estral nesta espécie. Foram utilizadas 18 éguas, totalizando 36 amostras de soro, sendo duas de cada égua. As amostras foram colhidas no estro e no diestro. As proteínas séricas totais foram obtidas pelo método do Biureto, a partir da utilização de Kits comerciais (LABTEST) e, as diferentes subfrações proteicas, por eletroforese em gel de poliacrilamida (SDS-PAGE). O eletroforetograma das proteínas séricas colocou em evidência a presença de 17 a 25 frações proteicas, cujos pesos moleculares variaram de 22 a 254 kDa. Identificaram-se duas proteínas ainda não nomeadas oficialmente, de massas moleculares (MM) 23 kDa e 144 kDa. Os valores médios ± SEM obtidos para cada variável no estro e no diestro, respectivamente, foram: proteínas totais (g/dL) 7,11 ± 0,07 e 7,36 ± 0,07; albumina (mg/dL) 4790,83 ± 69,10 e 5027,19 ± 69,10; α1 glicoproteína ácida (mg/dL) 4,90 ± 0,31 e 4,93 ± 0,31; ceruloplasmina (mg/dL) 15,28 ± 1,31 e 10,65 ± 1,31; haptoglobina (mg/dL) 22,70 ± 1,16 e 27,06 ± 1,16; transferrina (mg/dL) 329,00 ± 9,78 e 350,16 ± 9,78; IgA (mg/dL) 119,91 ± 6,30 e 107,03 ± 6,30; IgG (mg/dL) 1525,07 ± 40,18 e 1517,25 ± 40,18; MM 23 (mg/dL) 204,44 ± 8,61 e 219,79 ± 8,61; MM 144 (mg/dL) 22,13 ± 0,55 e 21,49 ± 0,55. Não houve diferença significativa das proteínas totais e suas frações do estro para o diestro. Conclui-se que as modificações hormonais durante as fases do ciclo estral da égua não interferem no proteinograma sérico.

This study aimed to obtain the electrophoretic profile of serum proteins in cyclic mares and to verify the differences between the follicular and luteal phases of the estrous cycle in this species. Eighteen mares were used, totaling 36 serum samples, two of each mare. Samples were collected both in estrus and in diestrus. Total serum proteins were obtained by the Biureto method, by using commercial kits (LABTEST), while the different protein subfractions by polyacrylamide gel electrophoresis (SDS-PAGE). The electroforetogram of serum proteins evidenced the presence of 17 to 25 protein fractions, whose molecular weights ranged from 22 to 254 kDa. Two proteins that were not yet officially named were identified, of molecular weights (MW) of 23 kDa and 144 kDa. The mean values (± SEM) obtained for each variable in estrus and diestrus were, respectively: total proteins (g/dL) 7.11 ± 0.07 and 7.36 ± 0.07; albumin (mg/dL) 4790.83 ± 69.10 and 5027.19 ± 69.10; α1 acid glycoprotein (mg/dL) 4.90 ± 0.31 and 4.93 ± 0.31; ceruloplasmine (mg/dL) 15.28 ± 1.31 and 10.65 ± 1.31; haptoglobine (mg/dL) 22.70 ± 1.16 and 27.06 ± 1.16; transferrin (mg/dL) 329.00 ± 9.78 and 350.16 ± 9.78; IgA (mg/dL) 119.91 ± 6.30 and 107.03 ± 6.30; IgG (mg/dL) 1525.07 ± 40.18 and 1517.25 ± 40.18; MW 23 (mg/dL) 204.44 ± 8.61 and 219.79 ± 8.61; MW 144 (mg/dL) 22.13 ± 0.55 and 21.49 ± 0.55. No significant difference was verified in total proteins and its fractions in estrus and diestrus. The hormonal changes during the specific stages of the estrous cycle of the mare do not interfere with the serum proteinogram.

Comparison between two different experimental models of osteoarthritis in rabbits. Intra-articular collagenase injection and anterior cruciate ligament transection.

PURPOSE:: To compare two different experimental models of osteoarthritis in rabbits: intra-articular collagenase injection and anterior cruciate ligament transection. METHODS:: Ten adult rabbits were randomly divided in two groups: COLL (collagenase group) and ACLT (anterior cruciate ligament transection). The COLL group was treated with 0.5 ml collagenase solution (2mg collagenase/0.5 ml sterile PBS), and the ACTL group was subjected to anterior cruciate ligament. After six and twelve weeks, respectively, the animals in the COLL and ACTL groups were euthanized. The gross appearance and histological examinations conducted in the cartilage articular surface was blindly scored according to the criteria developed by Yoshimi et al. (1994) and Mankin et al. (1971), respectively. RESULTS:: The gross morphologic observation, macroscopic score and histological examinations have demonstrated that the ACTL group presented the highest scores, and lesions more severe than those in the COLL group. CONCLUSIONS:: Both methods, anterior cruciate ligament transection and collagenase, applied to the stifle joint of the rabbits have effectively induced degenerative changes in the cartilage tissue, through statistically significant analysis (p≤0.05). The ACTL method has presented more severe lesions.

Comparison between two different experimental models of osteoarthritis in rabbits. Intra-articular collagenase injection and anterior cruciate ligament transection

ABSTRACT PURPOSE: To compare two different experimental models of osteoarthritis in rabbits: intra-articular collagenase injection and anterior cruciate ligament transection. METHODS: Ten adult rabbits were randomly divided in two groups: COLL (collagenase group) and ACLT (anterior cruciate ligament transection). The COLL group was treated with 0.5 ml collagenase solution (2mg collagenase/0.5 ml sterile PBS), and the ACTL group was subjected to anterior cruciate ligament. After six and twelve weeks, respectively, the animals in the COLL and ACTL groups were euthanized. The gross appearance and histological examinations conducted in the cartilage articular surface was blindly scored according to the criteria developed by Yoshimi et al. (1994) and Mankin et al. (1971), respectively. RESULTS: The gross morphologic observation, macroscopic score and histological examinations have demonstrated that the ACTL group presented the highest scores, and lesions more severe than those in the COLL group. CONCLUSIONS: Both methods, anterior cruciate ligament transection and collagenase, applied to the stifle joint of the rabbits have effectively induced degenerative changes in the cartilage tissue, through statistically significant analysis (p≤0.05). The ACTL method has presented more severe lesions.

Efeitos de diferentes doses de hiclato de doxiciclina nos parâmetros hematológicos de cães com erliquiose / Effects of different doses of doxycycline hyclate on haematological parameters of dogs with ehrlichiosis

Este estudo objetivou comparar os efeitos de duas doses de hiclato de doxiciclina sobre a contagem de hemácias, concentração de hemoglobina, hematócrito, contagem de leucócitos e plaquetas de cães com erliquiose. Foram estudados os grupos I, composto por cães saudáveis (n=6), negativos na sorologia para Ehrlichia canis e Leptospira spp., na PCR em tempo real para E. canis e Anaplasma platys e na semi–nested PCR para Babesia canis; grupos II (n=6) e III (n=6), compostos por cães com história clínica sugestiva, sorologia e/ou PCR em tempo real positivas para E. canis, e negativos na pesquisa de anticorpos anti-Leptospira spp., na PCR em tempo real para A. platys e na semi–nested PCR para B. canis. Os cães doentes foram tratados com hiclato de doxiciclina via oral a cada 12 horas por 30 dias (5 mg/kg, grupo II; 10 mg/kg, grupo III). Hemogramas foram feitos antes, decorridos 15 dias e aos 10 dias após o término dos tratamentos. Não houve diferenças entre os grupos nos tempos avaliados quanto às variáveis hemácias, hemoglobina, hematócrito e leucócitos. Observou-se diferença quanto aos trombócitos entre o grupo I e os grupos II e III (p<0,0001) no início do estudo. Aos 15 dias de tratamento, a média do grupo III foi menor que dos grupos I (p=0,008) e II (p=0,0007), indicando persistência da trombocitopenia, ausente no grupo II. Não houve diferença entre os grupos no tempo final, sugerindo que ambos os tratamentos promoveram aumento da contagem de plaquetas em cães naturalmente infectados por E. canis.

This study aimed to compare the effects of two doses of doxycycline hyclate on red blood cells, hemoglobin, hematocrit, white blood cells and platelets of dogs with ehrlichiosis. Group I, comprised of healthy dogs (n=6), negative on serology for Ehrlichia canis and Leptospira spp., real time PCR forE. canis and Anaplasma platys, and on semi–nested PCR for Babesia canis; Groups II (n=6) and III(n=6), comprised of dogs with suggestive clinical history, positive serology and/or real time PCR for E.canis, negative on research for anti-Leptospira spp. antibodies and real time PCR for A. platys, and on semi–nested PCR for B. canis were studied. Sick dogs were treated with doxycycline hyclate every 12hours, by mouth, for 30 days (5 mg/kg, group II; 10 mg/kg, group III). Complete blood counts were performed before, after 15 days, and 10 days after period of treatment was complete. No difference between groups at the studied time points were noticed for red blood cells, hemoglobin, haematocrit and white blood cells. Difference was observed for platelets between group I and groups II and III(p<0.0001) at the study on set. After 15 days of treatment, the mean platelet for group III was lower than groups I (p=0.008) and II (p=0.0007), indicative of persistent thrombocytopenia, already absentin group II. No difference between groups was noticed at final time point, which suggests that both treatments increased platelets in dogs naturally infected with E. canis.

Proteinograma do líquido sinovial de equinos hígidosobtido por eletroforese em gel de poliacrilamida / Proteinogram of synovial liquid from healthy equines determined by means of sodium dodecyl sulphate-polyacrylamide gel electrophoresis

O objetivo do presente trabalho foi determinar o proteinograma do líquido sinovial de quarenta equinos hígidos. Após colheita, centrifugação e fracionamento das amostras, as proteínas de fase aguda foram separadas por eletroforese em gel de poliacrilamida, contendo SDS-PAGE, e suas concentrações determinadas por densitometria computadorizada. Foram encontradas 16 a 26 proteínas no fracionamento eletroforético, compeso molecular variando de 16 a 238.000 Daltons (Da). Os pesos moleculares encontrados, correspondentes às proteínas mais conhecidas, foram albumina, 63.000Da; ceruloplasmina, 113.000Da; transferrina, 80.000Da; haptoglobina, 40.000Da; α-1-glicoproteínaácida, 37.000Da; IgA, 189.000Da e IgG, 74.000Da. O estabelecimento do proteinograma do líquido sinovial de equinos hígidos pode trazer contribuição significativa ao diagnóstico, tratamento e prognóstico de animais com alterações articulares.

The aim of this study was to determine the proteinogram of the proteins included in the synovial fluid of forty healthy horses. After collection, centrifugation and fractionation of the samples, the acute phase proteins were separated by electrophoresis in polyacrylamide gel containing SDS-PAGE, and their concentrations were determined by computerized densitometry. It was found 16 to 26 proteins on the electrophoretic fraction, with molecular weights from 16 to 238.000 Daltons (Da). The molecular weights found, corresponding to most known proteins were albumin, 63,000Da; ceruloplasmin, 113.000Da; transferrin, 80,000Da; haptoglobin, 40.000Da, α-1-acid glycoprotein, 37.000Da; IgA, IgG and 189.000Da, 74.000Da. The knowledge proteinogram of synovial fluid of healthy horses can bring significant contribution to the diagnosis, treatment and prognosis of animals with articular changes.

False positivity of gamma-glutamyl transpeptidase measurement in urine.

Although enzymuria tends to be associated to renal injury, there are no studies that have evaluated the presence of the enzyme gamma-glutamyl transpeptidase (GGT) spectrophotometry in the urine using a non-nephrotoxic agent (Nerium oleander) in order to evaluate the possibility of false positive results. The urinary GGT/urinary creatinine concentration ratio (uGGT/uCr) of 10 healthy dogs was calculated and posteriorly confronted with data from clinical evaluation, hematological and serum biochemical profiles, creatinine clearance (CrC), urinalysis, urine protein/creatinine ratio (UPC), electrocardiogram, systemic blood pressure (SBP) and light and electron microscopy. The results for kidney histology, SBP, UPC and CrC were not significantly different in any of the time-points analyzed. However, uGGT/uCr was significantly higher when measured 4 hours and 24 hours after administration of N. oleander. The measurement of the urinary GGT enzyme, as performed in many studies, yielded false positive results in dogs poisoned by a non-nephrotoxic agent.

Proteinograma sérico e do líquido peritoneal de equinos hígidos e daqueles submetidos à obstrução intestinal experimental / Serum and peritoneal protein profile of healthy horses and horses subjected to experimental intestinal obstruction

Avaliou-se a resposta de fase aguda através da concentração das proteínas de fase aguda (PFA) no soro sanguíneo e no líquido peritoneal de vinte e um equinos, hígidos e submetidos à obstrução intestinal experimental, distribuídos em quatro grupos: obstrução de duodeno - GD (n=6), íleo - GI (n=6), cólon dorsal esquerdo - GM (n=6) e controle instrumentado - GC (n=3). Foram colhidas amostras de sangue e líquido peritoneal e, após centrifugação e fracionamento, as proteínas de fase aguda foram separadas por eletroforese em SDS-PAGE. Identificaram-se as proteínas IgA, ceruloplasmina, transferrina, albumina, IgG, haptoglobina, α1-glicoproteína ácida e P24, no soro e no líquido peritoneal. Houve aumento nas concentrações sérica e peritoneal de todas as PFA, sendo mais evidente no líquido peritoneal e nos animais obstruídos. O fracionamento eletroforético das PFA no líquido peritoneal é mais eficaz no diagnóstico de processos inflamatórios abdominais, quando comparado ao sérico.

It was evaluated the acute phase response over the concentration of acute phase proteins (APP) in serum and in peritoneal fluid of twenty-one healthy equines submitted to experimental intestinal obstruction distributed in groups: duodenal obstruction (GD), ileum obstruction (GI), left-dorsal colon (GM), and instrumentalized control (GC). Blood and peritoneal liquid samples were collected, after centrifugation and fractioning and the acute phase proteins were separated by electrophoresis in SDS-PAGE. The acute phase proteins (APP) identified were IgA, ceruloplasmin, transferrin, albumin, IgG, haptoglobin, α1-acid glycoprotein and P24, in the serum and liquid peritoneal. There was anincrease in the serum and peritoneal proteins concentrations evidencing a higher response of the APP from the peritoneal fluid and of the obstructed groups. The electrophoretic separation of the proteins included in the peritoneal liquid is more effective when compared to the serum concentration in the diagnosis of abdominal inflammatory processes.

Comparative study between fibrin glue and platelet rich plasma in dogs skin grafts.

PURPOSE: Compare fibrin glue (Tissucol(®)) and platelet-rich plasma in full-thickness mesh skin grafts in dogs. METHODS: Eighteen dogs were used, divided into two groups: fibrin glue (FG) and platelet-rich plasma (PRP). In all the animals, a full-thickness 3x3 cm mesh skin graft was implanted. In the left limb, the biomaterial was place between the graft and the receptor bed, according to the group, while the right limb served as the control group. All the animals were evaluated clinically every 48 hours until the 14th day, using the variables of exudation, coloration, edema and cosmetic appearance. Three animals were evaluated histologically, on the third, seventh and tenth postoperative days, using the variables of fibroblasts, collagen, granulation tissue, microscopic integration-adherence and acute inflammation. RESULTS: Clinical evaluations showed that the group CF showed better scores for all variables compared to PRP group. On the histological evaluations PRP group had a higher presence of fibroblasts in the seventh and fourteenth days. CONCLUSION: The fibrin glue group was clinically superior to the platelet-rich group when used on full-thickness skin grafts in dogs.

Comparative study between fibrin glue and platelet rich plasma in dogs skin grafts / Estudo comparativo entre a cola de fibrina e o plasma rico em plaquetas em enxertos cutâneos em cães

PURPOSE: Compare fibrin glue (Tissucol®) and platelet-rich plasma in full-thickness mesh skin grafts in dogs. METHODS: Eighteen dogs were used, divided into two groups: fibrin glue (FG) and platelet-rich plasma (PRP). In all the animals, a full-thickness 3x3 cm mesh skin graft was implanted. In the left limb, the biomaterial was place between the graft and the receptor bed, according to the group, while the right limb served as the control group. All the animals were evaluated clinically every 48 hours until the 14th day, using the variables of exudation, coloration, edema and cosmetic appearance. Three animals were evaluated histologically, on the third, seventh and tenth postoperative days, using the variables of fibroblasts, collagen, granulation tissue, microscopic integration-adherence and acute inflammation. RESULTS: Clinical evaluations showed that the group CF showed better scores for all variables compared to PRP group. On the histological evaluations PRP group had a higher presence of fibroblasts in the seventh and fourteenth days. CONCLUSION: The fibrin glue group was clinically superior to the platelet-rich group when used on full-thickness skin grafts in dogs.

OBJETIVO: Comparar a cola de fibrina (Tissucol®) e o plasma rico em plaquetas em enxertos cutâneos de espessura completa em malha em cães. MÉTODOS: Foram utilizados 18 cães, distribuídos em dois grupos, cola de fibrina (CF) e plasma rico em plaquetas (PRP). Em todos os animais foi realizado um enxerto cutâneo de 3x3 cm, em malha de espessura completa. No membro esquerdo foi colocado o biomaterial entre o enxerto e o leito receptor, cada qual em seu grupo, o membro direito serviu como grupo controle. Todos os animais foram avaliados clinicamente a cada 48 horas até o décimo quarto dia, através das variáveis: exsudação, coloração, edema e aspecto cosmético; histologicamente em três animais, no terceiro, sétimo e décimo quarto dia de pós-operatório através das variáveis: fibroblastos, colágeno, tecido de granulação, integração-aderência microscópica e inflamação aguda. RESULTADOS: Avaliações clínicas demonstraram que o grupo CF apresentou melhor escores em todas variáveis quando comparado com o grupo PRP. Nas avaliações histológicas o grupo PRP apresentou maior presença de fibroblastos ao sétimo e décimo quarto dia. CONCLUSÃO: A cola de fibrina foi clinicamente superior ao grupo plasma rico em plaquetas quando usados em enxertos cutâneos de espessura completa em cães.

Experimental Ehrlichia canis infection changes acute-phase proteins.

Early diagnosis of canine ehrlichiosis favors prompt institution of treatment and improves the prognosis for the animal, since this disease causes mortality among dogs. Studies have shown that determining the concentration of acute-phase proteins (APPs) may contribute towards early detection of disease and aid in predicting the prognosis. This study aimed to evaluate the APP profile in dogs experimentally infected with Ehrlichia canis, at the start of the infection and after treatment. It also investigated whether any correlation between APP levels and the clinical and laboratory alterations over the course of the disease would be possible. The results obtained showed abnormal levels of all the APPs on the third day after infection (D3), with the highest levels being reached on D18, with the exception of ceruloplasmin and acid glycoprotein, which presented their peaks on D6 and D12 respectively. We concluded that assessment of APP levels could contribute towards establishing an early diagnosis of canine ehrlichiosis, particularly regarding acid glycoprotein and ceruloplasmin, since these proteins were detected at increased levels even before the onset of clinical and laboratory findings of the disease.

Experimental Ehrlichia canis infection changes acute-phase proteins / A infecção experimental por Ehrlichia canis altera as proteínas de fase aguda

Early diagnosis of canine ehrlichiosis favors prompt institution of treatment and improves the prognosis for the animal, since this disease causes mortality among dogs. Studies have shown that determining the concentration of acute-phase proteins (APPs) may contribute towards early detection of disease and aid in predicting the prognosis. This study aimed to evaluate the APP profile in dogs experimentally infected with Ehrlichia canis, at the start of the infection and after treatment. It also investigated whether any correlation between APP levels and the clinical and laboratory alterations over the course of the disease would be possible. The results obtained showed abnormal levels of all the APPs on the third day after infection (D3), with the highest levels being reached on D18, with the exception of ceruloplasmin and acid glycoprotein, which presented their peaks on D6 and D12 respectively. We concluded that assessment of APP levels could contribute towards establishing an early diagnosis of canine ehrlichiosis, particularly regarding acid glycoprotein and ceruloplasmin, since these proteins were detected at increased levels even before the onset of clinical and laboratory findings of the disease.

O diagnóstico precoce da erliquiose canina favorece a pronta instituição do tratamento e melhora o prognóstico do animal, pois se trata de uma doença de alta mortalidade em cães. Estudos têm apontado que a determinação da concentração de proteínas de fase aguda (PFA) pode contribuir para detecção precoce de doenças e auxiliar na predição do prognóstico. O presente estudo objetivou avaliar o perfil de proteínas de fase aguda (PFA) em cães experimentalmente infectados com Ehrlichia canis, no início da infecção e após o tratamento. Além disso, se seria possível associá-las com as alterações clínico-laboratoriais durante o curso da doença. Os resultados obtidos evidenciaram que todas as PFA estudadas alteraram suas concentrações em D3 (dia 3), comparadas ao D0, atingindo concentrações máximas em D18, com exceção da ceruloplasmina e da glicoproteína ácida, cujos picos foram observados em D6 e D12, respectivamente. Concluímos que a avaliação das concentrações de PFA poderiam contribuir para o diagnóstico precoce da erliquiose canina, principalmente com relação à ceruloplasmina e glicoproteína ácida, pois seus aumentos foram anteriores ao aparecimento dos sinais clínicos e das alterações laboratoriais da doença.