Isolation of brucella spp from milk of brucellosis positive cows in São Paulo and Minas Gerais states

A brucelose é uma zoonose crônica de importância para a Saúde Pública. Considerando o pequeno número de dados brasileiros sobre a sua presença em leite cru e derivados não-pasteurizados, estudamos a presença de brucelas em leite de animais sorologicamente positivos. A soroaglutinação rápida (SAR), a soroaglutinação lenta (SAL) e a soroaglutinação lenta com tratamento do soro com 2-mercaptoetanol foram utilizados para identificar os animais positivos nas propriedades estudadas. Amostras diárias de 300 ml de leite foram colhidas por três dias de todos os quartos mamários produtivos (75 ml/teto). As amostras eram misturadas e centrifugadas. Parte do sedimento e do sobrenadante foi inoculada em meios de Farrel e Brodie-Sinton (BS) suplementados com agentes antimicrobianos. As placas e tubos foram cultivados por sete dias a 37ºC, em microaerofilia. As colônias suspeitas no meio BS foram imediatamente repicadas para ágar-Brucella, e cultivadas sob a mesma condição. Os microrganismos isolados foram submetidos a procedimentos de identificação, incluindo a coloração de Gram, requerimento de CO2, produção de H2S, atividade da urease e crescimento na presença de tionina e fucsina. Das 49 amostras examinadas, isolou-se Brucella abortus de 15 (30,61%). Os biótipos isolados foram: biótipo 1 em uma amostra (2,04%), biótipo 2 em oito (16,32%) e biótipo 3 em seis amostras (12,25%)

Isolation and immunophenotypic characterization of mesenchymal stem cells derived from equine species adipose tissue

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Effects of buprenorphine, carprofen and saline on thermal and mechanical nociceptive thresholds in cats

To evaluate a prototype pressure stimulus device for use in the cat and to compare with a known thermal threshold device.Eight healthy adult cats weighing between 3.0 and 4.9 kg.Pressure stimulation was given via a plastic bracelet taped around the forearm. Three 2.4 mm diameter ball bearings, in a 10-mm triangle, were advanced against the craniolateral surface of the antebrachium by manual inflation of a modified blood pressure bladder. Pressure in the cuff was recorded at the end point (leg shake and head turn). Thermal threshold was also tested. Stimuli were stopped if they reached 55 degrees C or 450 mmHg without response. After four pressure and thermal threshold baselines, each cat received SC buprenorphine 0.01 mg kg(-1), carprofen 4 mg kg(-1) or saline 0.3 mL in a three period cross-over study with a 1-week interval. The investigator was blinded to the treatment. Measurements were made at 0.25. 0.5, 0.75, 1, 2, 3, 4, 6, 8, and 24 hours after injection. Data were analyzed by using ANOVA.There were no significant changes in thermal or pressure threshold after administration of saline or carprofen, but thermal threshold increased from 60 minutes until 8 hours after administration of buprenorphine (p < 0.05). The maximum increase in threshold from baseline (Delta T-max) was 3.5 +/- 3.1 degrees C at 2 hours. Pressure threshold increased 2 hours after administration of buprenorphine (p < 0.05) when the increase in threshold above baseline (Delta P-max) was 162 +/- 189 mmHg.This pressure device resulted in thresholds that were affected by analgesic treatment in a similar manner but to a lesser degree than the thermal method. Pressure stimulation may be a useful additional method for analgesic studies in cats.

Prevalence of gastric lesions (ulcers and/or erosions) and their relationship to possible stressfull factors in asymptomatic Quarter Horse foals: endoscopic survey

A úlcera gástrica constitui-se numa das mais importantes causas de desconforto abdominal em eqüinos jovens. Com o objetivo de se verificar a prevalência de lesões gástricas (úlceras e/ou erosões) e sua relação com fatores como estresse, idade, e sexo, sessenta potros da raça Quarto de Milha não portadores de sinais clínicos compatíveis com lesões gástricas foram submetidos à gastroscopia. Os potros foram divididos em quatro faixas etárias de 15 animais cada uma, sendo: 1 a 30 dias, 31 a 60 dias, 61 a 90 dias e 91 a 120 dias de idade. A prevalência de lesões gástricas foi de 43,3%. Animais com idade entre 61 a noventa dias foram os mais acometidos. Não houve diferença significativa entre as faixas etárias. Descamações do epitélio aglandular ocorreram em nove potros (60%) entre um e trinta dias, em seis (40%) entre 31 e sessenta dias e em apenas dois (6,6%) com idade superior a sessenta dias. Fatores considerados estressantes, como infestação intensa por carrapatos (29), problemas respiratórios (3), dermatopatias (3), babesiose (2), onfaloflebite (1), diarréia (1), problemas ortopédicos (1) e ferida lacerante com presença de miíase (1) não influenciaram a ocorrência das lesões. Machos e fêmeas foram igualmente acometidos.

Lesions (ulcers and/or erosions) and desquamations location in the gastric mucosa from asymptomatic Quarter Horse foals: endoscopic survey

A úlcera gástrica figura como uma importante causa de desconforto abdominal em eqüinos jovens. de acordo com a localização das lesões na mucosa gástrica, a presença ou ausência de sinais clínicos e possíveis complicações resultantes de sua ocorrência, quatro síndromes clínicas são freqüentemente descritas em potros: 1) Úlceras assintomáticas ou silenciosas; 2) Úlceras sintomáticas ou ativas; 3) Úlceras perfuradas; e 4) Obstruções gástrica ou duodenal. Com o objetivo de se verificar a distribuição de lesões gástricas (úlceras e/ou erosões) e descamações do epitélio aglandular no estômago de eqüinos jovens assim como uma possível relação entre as alterações mencionadas (lesão/descamação) sessenta potros da raça Quarto de Milha não-portadores de sinais clínicos compatíveis com úlceras gástricas foram submetidos à gastroscopia. Os potros foram divididos em quatro faixas etárias de 15 animais cada uma, sendo: 1 a 30 dias, 31 a 60 dias, 61 a 90 dias e 91 a 120 dias de idade. As lesões ocorreram em ordem decrescente de freqüência nas regiões aglandular próximo ao margo plicatus ao longo da curvatura maior, aglandular próximo à cárdia ao longo da curvatura menor, fundo glandular e aglandular e antro. As descamações do epitélio aglandular ocorreram de forma similar nas regiões de fundo e margo plicatus. Não houve associação entre a ocorrência de lesões e descamações.

Effects of methadone, alone or in combination with acepromazine or xylazine, on sedation and physiologic values in dogs

Objective To evaluate the effects of methadone, administered alone or in combination with acepromazine or xylazine, on sedation and on physiologic values in dogs.Study design Randomized cross-over design.Animals Six adult healthy mixed-breed dogs weighing 13.5 +/- 4.9 kg.Methods Dogs were injected intramuscularly with physiologic saline (Control), or methadone (0.5mg kg(-1)) or acepromazine (0.1 mg kg(-1)) or xylazine (1.0 mg kg(-1)), or acepromazine (0.05 mg kg(-1)) plus methadone (0.5 mg kg(-1)) or xylazine (0.5 mg kg(-1)) plus methadone (0.5 mg kg(-1)) in a randomized cross-over design, with at least 1-week intervals. Sedation, pulse rate, indirect systolic arterial pressure, respiratory rate (RR), body temperature and pedal withdrawal reflex were evaluated before and at 15-minute intervals for 90 minutes after treatment.Results Sedation was greater in dogs receiving xylazine alone, xylazine plus methadone and acepromazine plus methadone. Peak sedative effect occurred within 30 minutes of treatment administration. Pulse rate was lower in dogs that received xylazine either alone or with methadone during most of the study. Systolic arterial pressure decreased only in dogs receiving acepromazine alone. When methadone was administered alone, RR was higher than in other treatments during most of the study and a high prevalence of panting was observed. In all treatments body temperature decreased, this effect being more pronounced in dogs receiving methadone alone or in combination with acepromazine. Pedal withdrawal reflex was absent in four dogs receiving methadone plus xylazine but not in any dog in the remaining treatments.Conclusions Methadone alone produces mild sedation and a high prevalence of panting. Greater sedation was achieved when methadone was used in combination with acepromazine or xylazine. The combination xylazine-methadone appears to result in better analgesia than xylazine administered alone. Both combinations of methadone/sedative were considered effective for premedication in dogs.

Effects of meperidine or saline on thermal, mechanical and electrical nociceptive thresholds in cats

Objective To measure cutaneous electrical nociceptive thresholds in relation to known thermal and mechanical stimulation for nociceptive threshold detection in cats.Study design Prospective, blinded, randomized cross-over study with 1-week washout interval.Animals Eight adult cats [bodyweight 5.1 +/- 1.8 kg (mean + SD)].Methods Mechanical nociceptive thresholds were tested using a step-wise manual inflation of a modified blood pressure bladder attached to the cat's thoracic limb. Thermal nociceptive thresholds were measured by increasing the temperature of a probe placed on the thorax. The electrical nociceptive threshold was tested using an escalating current from a constant current generator passed between electrodes placed on the thoracic region. A positive response (threshold) was recorded when cats displayed any or all of the following behaviors: leg shake, head turn, avoidance, or vocalization. Four baseline readings were performed before intramuscular injection of meperidine (5 mg kg(-1)) or an equal volume of saline. Threshold recordings with each modality were made at 15, 30, 45, 60, 90, and 120 minutes post-injection. Data were analyzed using ANOVA and paired t-tests (significance at p < 0.05).Results There were no significant changes in thermal, mechanical, or electrical thresholds after saline. Thermal thresholds increased at 15-60 minutes (p < 0.01) and mechanical threshold increased at 30 and 45 minutes after meperidine (p < 0.05). Maximum thermal threshold was +4.1 +/- 0.3 degrees C above baseline at 15 minutes while maximum mechanical threshold was 296 +/- 265 mmHg above baseline at 30 minutes after meperidine. Electrical thresholds following meperidine were not significantly different than baseline (p > 0.05). Thermal and electrical thresholds after meperidine were significantly higher than saline at 30 and 45 minutes (p < 0.05), and at 120 minutes (p < 0.05), respectively. Mechanical thresholds were significantly higher than saline treatment at 30 minutes (p <= 0.05).Conclusion and clinical relevance Electrical stimulation did not detect meperidine analgesia whereas both thermal and mechanical thresholds changed after meperidine administration in cats.

Comparison of the cardio-respiratory effects of methadone and morphine in conscious dogs

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Electroacupuncture analgesia in dogs: is there a difference between uni- and bi-lateral stimulation?

Objective To compare the analgesic effect of uni- and bi-lateral electroacupuncture (EA) in response to thermal and mechanical nociceptive stimuli and to investigate the cardiorespiratory, endocrine, and behavioral changes in dogs submitted to EA.Study design Prospective, randomized cross-over experimental study.Animals Eight adult, clinically healthy, cross-breed dogs, weighing 13 +/- 4 kg.Methods Dogs underwent electrostimulation at false acupoints (T-false); bilateral EA at acupoints, stomach 36, gall bladder 34 and spleen 6 (T-EA/bil); unilateral EA at the same points (T-EA/uni) or were untreated (T-control). All animals received acepromazine (0.05 mg kg(-1)) IV; and heart rate, pulse oximetry, indirect arterial blood pressure, respiratory rate, PECO2, rectal temperature, and plasma cortisol concentration were measured before, during, and after EA. Analgesia was tested using thoracic and abdominal cutaneous thermal and mechanical stimuli, and an interdigital thermal stimulus. Behavior was classified as calm or restless. Analysis of variance for repeated measures followed by Tukey's test was used for analysis of the data.Results There were no cardiorespiratory differences among the treatments. The cutaneous pain threshold was higher after EA, compared with false points. The latency period was shorter and analgesia was more intense in T-EA/bil than T-EA/uni, when both were compared with T-false and T-control. Six out of eight animals treated with EA were calm during treatment, and 5/8 and 4/8 of the T-false and T-control animals, respectively, were restless. Latency to interdigital thermal stimulation increased in T-EA/bil compared with the others. There was no difference in plasma cortisol concentrations among the treatments.Conclusions Bilateral EA produced a shorter latency period, a greater intensity, and longer duration of analgesia than unilateral stimulation, without stimulating a stress response.Clinical relevance Bilateral EA produces a better analgesic effect than unilateral EA.

Effect of intravenous propofol and remifentanil on heart rate, blood pressure and nociceptive response in acepromazine premedicated dogs

ObjectiveTo investigate the cardiorespiratory, nociceptive and endocrine effects of the combination of propofol and remifentanil, in dogs sedated with acepromazine.Study designProspective randomized, blinded, cross-over experimental trial.AnimalsTwelve healthy adult female cross-breed dogs, mean weight 18.4 +/- 2.3 kg.MethodsDogs were sedated with intravenous (IV) acepromazine (0.05 mg kg-1) followed by induction of anesthesia with IV propofol (5 mg kg-1). Anesthesia was maintained with IV propofol (0.2 mg kg-1 minute-1) and remifentanil, infused as follows: R1, 0.125 mu g kg-1 minute-1; R2, 0.25 mu g kg-1 minute-1; and R3, 0.5 mu g kg-1 minute-1. The same dogs were administered each dose of remifentanil at 1-week intervals. Heart rate (HR), mean arterial pressure (MAP), respiratory rate (f(R)), end tidal CO(2) (Pe'CO(2)), arterial hemoglobin O(2) saturation, blood gases, and rectal temperature were measured before induction, and 5, 15, 30, 45, 60, 75, 90, and 120 minutes after beginning the infusion. Nociceptive response was investigated by electrical stimulus (50 V, 5 Hz and 10 ms). Blood samples were collected for plasma cortisol measurements. Statistical analysis was performed by anova (p < 0.05).ResultsIn all treatments, HR decreased during anesthesia with increasing doses of remifentanil, and increased significantly immediately after the end of infusion. MAP remained stable during anesthesia (72-98 mmHg). Antinociception was proportional to the remifentanil infusion dose, and was considered satisfactory only with R2 and R3. Plasma cortisol concentration decreased during anesthesia in all treatments. Recovery was smooth and fast in all dogs.Conclusions and clinical relevanceInfusion of 0.25-0.5 mu g kg-1 minute-1 remifentanil combined with 0.2 mg kg-1 minute-1 propofol produced little effect on arterial blood pressure and led to a good recovery. The analgesia produced was sufficient to control the nociceptive response applied by electrical stimulation, suggesting that it may be appropriate for performing surgery.

Cloning, sequencing and expression analysis of the equine hepcidin gene by real-time PCR

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Extramedullary hematopoiesis in a case of benign mixed mammary tumor in a female dog: cytological and histopathological assessment

Backgroud: Extramedullary hematopoiesis (EMH) is defined as the presence of hematopoietic stem cells such as erythroid and myeloid lineage plus megakaryocytes in extramedullary sites like liver, spleen and lymph nodes and is usually associated with either bone marrow or hematological disorders. Mammary EMH is a rare condition either in human and veterinary medicine and can be associated with benign mixed mammary tumors, similarly to that described in this case.Case presentation: Hematopoietic stem cells were found in a benign mixed mammary tumor of a 7-year-old female mongrel dog that presents a nodule in the left inguinal mammary gland. The patient did not have any hematological abnormalities. Cytological evaluation demonstrated two distinct cell populations, composed of either epithelial or mesenchymal cells, sometimes associated with a fibrillar acidophilic matrix, apart from megakaryocytes, osteoclasts, metarubricytes, prorubricytes, rubricytes, rubriblasts, promyelocytes, myeloblasts. Histological examination confirmed the presence of an active hematopoietic bone marrow within the bone tissue of a benign mammary mixed tumor.Conclusions: EMH is a rare condition described in veterinary medicine that can be associated with mammary mixed tumors. It's detection can be associated with several neoplastic and non-neoplastic mammary lesions, i.e. osteosarcomas, mixed tumors and bone metaplasia.

Serological profile of Toxoplasma gondii and Neospora caninum infection in commercial sheep from São Paulo State, Brazil

Neosporosis and toxoplasmosis are two important infections in young and adult sheep, leading to low production and abortion. This study aimed to determine the frequency of antibodies to Toxoplasma gondii and Neospora caninum in sheep from the eastern region of São Paulo State, Brazil. Serum samples (382) were collected from the sheep and assayed for T. gondii through modified agglutination test (MAT) and indirect fluorescent antibody test (IFAT), and for N. caninum antibodies, through IFAT, with cut-off titers equal to 16 (T. gondii) and 25 (N. caninum). All frozen samples were sent to the Center for Zoonoses Research (NUPEZO), Department of Veterinary Hygiene and Public Health (DHSVP), FMVZ, UNESP, for serological tests. A total of 71/382(18.6%) samples reacted to T. gondii, especially at titers 16 (28; 39.4%), 64(15; 21.1%), 256 (21; 29.6%) and 1024 (6; 8.5%) by MAT, and 16(34: 47.9%), 64 (18; 25.4%), 256 (14; 19.7%) and 1024 (5; 7%) by IFAT. As regards N. caninum, 49/382 (12.8%) samples reacted at titers 25 (17; 34.7%), 50 (11; 22.5%), 100 (11; 22.5%), and >= 200 (10; 20.4%). These animals presented infection but no clinical signs. Six and ten animals had high titers for toxoplasmosis and neosporosis. No significant association was observed between antibodies for both parasites (P = 0.535) according to Fisher's exact test, and no correlation was found between T. gondii (MAT) and N. caninum antibody titers (r = -0.0068; P=0.895), T. gondii (IFAT) and N. caninum antibody titers (r = -0.0025; P = 0.961). Thus, T. gondii and N. caninum infections were observed in farms located in São Paulo State, where sheep play an important economical role for the national and regional business. (C) 2010 Elsevier By. All rights reserved.

Epidemiological aspects in leptospirosis. Research of anti-Leptospira spp antibodies, isolation and biomolecular research in bovines, rodents and workers in rural properties from Botucatu, SP, Brazil.

Leptospirosis is a worldwide infection, transmitted between man and animals that causes a decrease in the production of bovine flocks, and offer risks for public health, as an important zoonosis. The rodents are the main reservoirs of leptospires. It was studied 27 dairy farm properties located in or near from Botucatu-SP, Brazil. In these farms were collected blood and kidney samples from rodents, blood and urine samples from bovines and blood samples from the workers. The serology was performed with microscopic agglutination test (MAT). Samples of bovine urine and rodent kidneys were cultivated searching for leptospires isolation. The polymerase chain reaction (PCR) of the kidneys of the rodents was performed. In MAT, 46/ 140 (32.85%) bovine and 8/34 (23.53%) human sera samples were positive, respectively. In human samples, the serovar Brastilava (37.51%) presented the highest occurrence, while in bovines, the serovars Hardjo and Castellonis were most frequent, with 26.08% each one. All of the rodents were negatives in serology. No leptospire was isolated, and kidney samples were negative in PCR. In bovines, the dam water and the bad hygiene quality of milking process were considered important risks of infection in the affected properties (p<0.05), where other reproductive problems, except abortion, can be related. In other side, to human beings the drainage system was the most important risk factor in the studied properties. Thus, it was verified the necessity of an improvement in zoosanitary handling of the properties, mainly of water supply.

Enrofloxacin assay validation and pharmacokinetics following a single oral dose in chickens

The pharmacokinetics of enrofloxacin (ENRO), a fluoroquinolone antimicrobial agent, was studied in male broiler chickens (Cobb) after single oral administration of 10 mg of ENRO/kg b.w. A high-performance liquid chromatography-photodiode array detector (DAD) (HPLC-DAD) method was developed and validated and used for quantitation of ENRO and its major metabolite ciprofloxacin in plasma. The HPLC analyses were carried out using a cationic-octadecyl mixed column and 0.05 mol/L phosphate buffer (pH 2.5)/acetonitrile as mobile phase. The sample preparation of plasma consisted of the precipitation of proteins followed by solid phase extraction on cationic-octadecyl mixed cartridges. The method was validated considering linear range, linearity, selectivity, sensitivity, limit of detection (LOD), limit of quantitation (LOQ), intra- and inter-day precisions and accuracy. The LOD and LOQ for both fluoroquinolones were 60 and 200 ng/mL for plasma. The plasma concentration vs. time graph was characteristic of a two-compartment open model. The maximal plasma concentration of 1.5 +/- 0.2 mg/mL was achieved at 9 +/- 2 h. The elimination half-life and the mean residence time of ENRO were 1.5 +/- 0.2 and 15.64 h, respectively. The area under the concentration-time curve was calculated as 35 +/- 4 mg(.)h/mL.