Post-mortem detection of hemoplasmas (hemotropic Mycoplasma spp.) in South American fur seal (Arctocephalus australis) sampled in Rio Grande do Sul State, southern Brazil.

Hemotropic mycoplasmas are bacteria that attaches to erythrocytes surface, which some species presents zoonotic concerns. In the suborder Pinnipedia, genera Otaria and Arctocephalus are prominent in Brazil. This study investigated the occurrence of hemoplasmas in Arctocephalus sp. and Otaria flavescens found dead along the coast of a Southern Brazilian State. DNA from 135 spleen samples were extracted and subjected to conventional PCR protocols, targeting the 16 S rRNA and 23 S rRNA gene. Three (2.22 %) Arctocephalus australis were positive in the 16 S rRNA gene, and no samples amplified in the 23 S rRNA gene. Samples from this study clustered with Zalophus californianus and Arctocephalus tropicalis mycoplasmas on a Bayesian phylogenetic analysis. Genetic diversity analysis suggested distinct genotypes, indicating A. australis as a new host for hemoplasma, and also a potential putative novel hemoplasma genotype. These findings raises future awareness for pinnipeds conservation, and adds Mycoplasma spp. to be taken into consideration when clinically evaluating rescued animals.

Applying children's enuresis treatment with amitriptyline for canine post-spaying urinary incontinence: A pilot estriol-controlled randomized clinical trial.

Urinary incontinence due to urethral sphincter mechanism incompetence (USMI) affects up to 20% of bitches that undergo spaying surgery. Amitriptyline is a tricyclic antidepressant whose urinary retention is a reported side effect. This study aimed to assess the efficacy and safety of amitriptyline when compared to estriol orally. Fifteen bitches with a clinical diagnosis of post-spaying UI were evaluated during 60 days in a non-blinded randomized clinical trial. All patients were enrolled after clinical evaluation consisting of anamnesis, physical examination, and complementary exams (complete blood count, biochemical parameters, urinalysis, and abdominal ultrasound). The amitriptyline (AMT) group consisted of 8 bitches, which received the initial dose of 1 mg/kg every 12 h, whereas the estriol (EST) group consisted of 7 bitches which were initially treated with 1 mg/animal every 24 h. Patients underwent clinical evaluation at 7 days, and then at 21 and 60 days of treatment to assess safety and efficacy, as well as adjustments of dose when necessary. A urinary incontinence scale was used to assess the level of incontinence and therapeutic response to treatment. During the period of the study, estriol was fully effective in 71% of cases and amitriptyline in 62%. Both drugs proved safe in the medical treatment of USMI, with adverse effects such as somnolence (AMT, n = 5/8) and male attraction (EST, n = 1/7). The results support the amitriptyline recommendation as a substitute for estriol in USMI treatment.

Comparison of serum creatinine, point-of-care symmetric dimethylarginine and renal imaging with glomerular filtration rate measured by renal scintigraphy in healthy and early chronic kidney diseased cats.

The aim of this study was to evaluate routinely used tests to diagnose cats in early stages of chronic kidney disease (CKD) and to describe a model for evaluating these variables simultaneously. Apparently healthy cats were screened using serum creatinine (sCr), point-of-care symmetric dimethylarginine (POC SDMA), urinalysis, urine protein/creatinine ratio (UPC) and imaging evaluation. Those parameters were compared to glomerular filtration rate (GFR) assessed by renal scintigraphy. Forty-four cats were included and consisted of 14 (31.8%) healthy cats (absence of abnormalities in renal morphology and sCr less than 1.6 mg/dL), 20 (45.5%) cats classified as CKD I (presence of abnormalities in renal morphology and sCr less than 1.6 mg/dL) and ten (22.7%) as CKD II (sCr equal to or greater than 1.6 mg/dL, with or without abnormalities in renal morphology). A large number (40.9%) of apparently healthy cats presented reduction in GFR, which included half of CKD I patients. Point-of-care SDMA was not a good predictor for decreased GFR, nor was it correlated with the variables GFR and sCr. Glomerular filtration rate was significantly lower in CKD I and II groups in comparison with healthy cats, but there was no significant difference between the CKD I and II groups. Multivariate logistic regression model identified three variables that affected the odds of a cat having decreased GFR (< 2.5 mL/min/kg): sCr (OR = 18.3; p = 0.019; CI = 1.6-207.2), and the ultrasonographic findings 'reduced corticomedullary definition' (OR = 19.9; p = 0.022; CI = 1.6-254.0) and 'irregular contour' (OR = 65.6; p = 0.003; CI = 4.2-1038.2). Renal ultrasonography evaluation should always be considered for screening early CKD in apparently healthy cats.

Viability of erythrocytes in canine packed red blood cells stored in CPDA-1 is related to the presence of Mycoplasma haemocanis.

Hemotropic mycoplasmas are associated with subclinical disease in dogs and should be identified in blood donors. The objective was to investigate the presence and effect of M. haemocanis in units of packed red blood cells (pRBC) during storage. Canine donors (n = 10) were screened for M. haemocanis by quantitative real-time PCR. pRBCs were obtained from 5 hemoplasma negative dogs and 5 hemoplasma positive dogs. Each pRBC was aliquoted into two 100 mL transfer bags and stored at 4 °C. M. haemocanis loads and biochemical variables (pH, bicarbonate, potassium, sodium, chlorite, glucose, lactate, ammonia, PCV, and % hemolysis) were evaluated on days 1, 7, 18, and 29. M. haemocanis loads increased in pRBC from day 1-29 of storage. Glucose decreased and lactate increase faster in pRBC with M. haemocanis. This study contributes to understand hemoplasma metabolism and reinforces that dog donors should be tested for hemoplasmas.

Hematologic variables of free-living Leptodactylus luctator with and without hemoparasites and thrombidiform mites in southern Brazil.

It has been suggested that anuran amphibian parasites can cause clinical signs in situations of environmental imbalance. In the family Leptodactylidae, information about hematology is scarce, although these are well-known tools for the diagnosis and prognosis in clinical practice and potential bioindicators of environmental stress. The objective of this study is to describe Leptodactylus luctator hematology, to report the occurrence of hemoparasites and thrombidiform mites, and to compare the hematological variables under the presence and absence of these organisms. Ectoparasites and heparinized blood samples from 40 free-living specimens of L. luctator were collected for analysis. Hematologic variables and total plasma protein were compared between groups with and without hemoparasites and intradermal mites. As results, structures compatible with hemogregarines, Lankesterella sp., five morphotypes of Trypanosoma spp., microfilaria, Aegyptianella sp., an unidentified intraleukocytic hemoparasite, and frog erythrocytic virus (FEV) inclusion bodies were identified in the blood samples, besides Hannemania spp. intradermal mites. The hemoparasite occurrence was higher than previously reported in other anuran families and locations. Also, L. luctator has smaller red blood cells (RBCs) and white blood cells (WBCs), and a hyposegmentation of the neutrophil nucleus, when compared to many other amphibians. White blood cell, neutrophil, and monocyte counts were higher in animals parasitized by mites. There was no correlation between the number of parasitized RBCs and hematologic variables. This study provides anuran hematologic information, in addition to indicating a host reaction to infestation by Hannemania spp. mites, besides constituting the first record of the distribution of hemoparasites and intradermal mites in L. luctator of the study region.

Assessment of selegiline and trilostane combined therapy efficacy for canine pituitary-dependent hypercortisolism treatment: A pilot randomized clinical trial.

Canine pituitary-dependent hypercortisolism (PDH) management with trilostane usually demands lifelong therapy. The greater the dose needed, the greater the risk of side effects. Selegiline therapy has been previously described but not commonly used for PDH treatment. The present work aimed to assess the efficacy of selegiline and trilostane combined therapy for canine PDH treatment. Fifteen client-owned dogs diagnosed with spontaneous PDH were enrolled. The patients were treated with trilostane (Tri group, n = 8, initial dose of 0.5 mg/kg, PO, q12h), or with trilostane and selegiline (Tri + Sel group, n = 7, initial trilostane dose of 0.5 mg/kg, PO, q12h and selegiline 1 mg/kg, PO, q24h). Dogs underwent clinical examination, serum biochemical analysis, urinalysis, abdominal ultrasound, and eACTH and post-ACTH cortisol measurements on treatment days zero (D0), 30 (D30), 90 (D90), and 180 (D180). There was a lack of adverse effects due to the combined therapy. Both groups showed a similar clinical response and lower post-ACTH cortisol levels at the study's end. There was no significant difference in trilostane dosage at D180 between groups. There was no documented increase in either right or left adrenal gland thickness in the Tri + Sel group in contrast with patients in the Tri group. However, there was no statistical difference between the groups regarding eACTH at D0 and D180. Patients in the Tri + Sel group achieved better serum triglycerides control at the end of the study. The association of selegiline with trilostane might be a feasible therapy for canine PDH; however, its eventual advantages need larger studies.

Weaning at 30, 75 and 180 days: Comparison between immune responses of beef calves.

The interruption of the maternal-filial bond in young calves can alter the physiological responses due to increased stress at this moment. Therefore, we aimed to analyze the association between age at weaning and the immune responses of 64 beef calves weaned abruptly. The animals were distributed in three weaning (W) treatments: hyper-early (W30), early (W75) and conventional (W180). The treatments agree with the weaning ages used in cow-calf systems in South America. Blood samples were collected of all experimental animals on the day of weaning and days 1, 2 and 7 post-weaning day of the three treatments. Blood samples were used to analyze hematological variables, total plasma proteins (TPP), fibrinogen, serum amyloid A protein (SAA) and cortisol. The variables analyzed were influenced by the sampling day (P < 0.001) and the calf gender (P < 0.05), and an interaction between treatment x sampling day was observed. Over the study period, high serum cortisol levels were observed for W30 calves. The serum amyloid A protein was significantly influenced by sampling day. When compared to W180 calves, W30 and W75 animals showed prolonged concentrations in plasma fibrinogen. In addition, they presented neutrophilia and lymphopenia during the 180-187 days period, which resulted in a greater neutrophil:lymphocyte ratio (N:L ratio). Calf gender resulted in changes in the number of blood leukocytes (P < 0.05). This result provides an alert for weaning at an early age, indicating that different management strategies must be tested to minimize physiological instabilities in critical periods of the life of animals.

Oral Acute and Repeated-Doses Toxicity Study of Valepotriates from Valeriana glechomifolia (Meyer) in Mice.

BACKGROUND: Species of Valeriana show sedative, hypnotic, anxiolytic, antidepressant and anti-inflammatory properties, which are associated with valepotriates. However, data about toxicity and safety of these compounds are still limited. The aim of this study was to investigate the toxicity of a valepotriate-enriched fraction (VAL) from Valeriana glechomifolia Meyer based on the Organization for Economic Cooperation and Development (OECD) guidelines 423 and 407. METHODS: In the acute study, CF1 mice were treated with a single dose of VAL (2000 mg/kg, p.o.) and observed for 14 days. In the repeated dose study, CF1 mice received single daily doses of VAL (30, 150 or 300 mg/kg, p.o.) or vehicle for 28 days. These doses were chosen based on previous results by our group and according to Guideline 407- OECD. RESULTS: The acute study allowed to classify VAL in the hazard category 5. The repeat-dose study has shown that VAL 300 mg/kg delayed weight gain and reduced food consumption in the first week, probably due to transient sedative effects. The other doses had no effect on animals' ponderal evolution. At the end of the treatment, all groups had equal body weight and food consumption. None of the doses altered any behavioral, urinary, biochemical, hematological, anatomic or histological parameters. CONCLUSION: A valepotriate-enriched fraction from Valeriana glechomifolia presents relatively low oral acute toxicity and does not induce evident toxicity after oral repeated treatment (at least up to 300 mg/kg) in mice.

Altered hematological and immunological parameters in silver catfish (Rhamdia quelen) following short term exposure to sublethal concentration of glyphosate.

Using agrichemicals to control unwanted species has become a necessary and common worldwide practice to improve crop production. Although most currently used agrichemicals are considered relatively safe, continuous usage contributes for soil and water contamination and collateral toxic effects on aquatic species. Few studies correlated the presence of agrichemicals on fish blood cells and natural immune system. Thus, in this study, silver catfish (Rhamdia quelen) were exposed to sublethal concentrations (10% of the LC(50-96 h)) of a glyphosate based herbicide and hematological and natural immune system parameters were evaluated. Silver catfish fingerlings exposed to glyphosate for 96 h had a significant reduction on blood erythrocytes, thrombocytes, lymphocytes and total leukocytes in contrast to a significant increase in the number of immature circulating cells. The effect of glyphosate on natural immune system was evaluated after 24h or 10 days exposure by measuring the phagocytic index of coelomic cells, and lysozyme, total peroxidase, bacteria agglutination, bactericidal activity and natural complement hemolytic activity in the serum of fingerlings. A significant reduction on phagocytic index, serum bacteria agglutination and total peroxidase was observed only after 24h exposure to glyphosate. In contrast, fingerlings exposed to glyphosate for 10 days had a significant lower serum bacteria agglutination and lysozyme activity. Glyphosate had no effect on serum bactericidal and complement natural hemolytic activity after 24h or 10 days exposure. Nonetheless, the information obtained in this study indicates that glyphosate contaminated water contributes to alter blood cells parameters and to reduce the activity of natural immune components important to mediate fish resistance to infecting microorganisms.

Análise comparativa entre microbiologia convencional, ELISA e PCR para detecção de Salmonella enteritidis, S. typhimurium, S. gallinarum e S. pullorum em carne de frango contaminada artificialmente

O presente trabalho teve como objetivo realizar uma análise comparativa das técnicas de Reação em Cadeia pela Polimerase(PCR) e Ensaio Imunoenzimático (ELISA SALVIA®) com o método microbiológico convencional para detecção e SalmonellaEnteritidis (SE), S. typhimurium (ST), S. gallinarum (SG) e S. pullorum (SP) em carne de frango. As amostras foram contaminadasartificialmente com diluições de 10-7, 10-8 e 10-9 para SE e ST e de 10-4, 10-5 e 10-6 para SG e SP, com cinco repetições decada diluição, totalizando 300 análises. Os testes foram realizados em cinco diferentes laboratórios para a validação dastécnicas. Na avaliação geral dos dados obtidos, a microbiologia convencional obteve 56,67% (170/300) de recuperação dasamostras contaminadas artificialmente, enquanto as técnicas de ELISA e PCR representaram 71% (213/300) e 75% (225/300), respectivamente. A análise dos resultados de detecção de Salmonella através dos testes ELISA e PCR, em relação aomicrobiológico convencional, apresentaram diferença estatística (p=0,0001, teste de MacNemar). Não houve diferença significativaentre os resultados da PCR e do ELISA. Os resultados alcançados demonstraram que, comparado ao microbiológicoconvencional, tanto o ELISA quanto a PCR foram eficazes para detecção dos sorovares de Salmonella nas amostras de carnede frango avaliadas.