Taxonomy of Physaloptera mirandai (Nematoda: Physalopteroidea) based in three-dimensional microscopy and phylogenetic positioning.

Gastrointestinal nematodes are important ecological assets for the maintenance of the biodiversity in the Atlantic Forest in Brazil. They parasitize a number of animals of the local fauna, in which some species can promote serious injuries in the stomach wall of their hosts, which may lead to death. Among these nematodes, parasites of the genus Physaloptera are known to parasitize mammals (particularly carnivores and small rodents), birds and reptiles, being important for the local biodiversity. In this work, three hundred and sixty-two nematodes were recovered from the stomach of twenty-one Metachirus nudicaudatus (Didelphimorphia: Didelphidae) collected in Duas Bocas Biological Reserve, State of Espírito Santo, one of the largest Atlantic Forest remnants and important wildlife refuge of the Atlantic Forest in Brazil. Analysis using fluorescence and scanning electron microscopy as well as phylogenetic assessment using the mitochondrial cytochrome c oxidase subunit I gene showed that the parasites belong to the Physaloptera. Our results show details of the nematode morphology including the cloacal papillae distribution, cuticular topography details, 2D and 3D measurements of the structures with taxonomic importance. Molecular data confirmed the validity of P. mirandai and the phylogeny supported the monophyly of the assemblage formed by Physaloptera and Turgida. The use of a combination of quantitative and multidimensional microscopy tools, such as 3D reconstruction and modeling, allied to phylogenetic analysis may provide grounds for a new approach on helminth taxonomy and structural characterization.

SHORT-COMMUNICATION Evaluation of perfused bovine udder for gene expression studies in dairy cows.

Intramammary infections are one of the main causes of productivity loss in dairy cows. To better understand the immune system response and to avoid the use of live animals, we validated the use of isolated bovine udder as an ex situ model. Six mammary glands were collected from cows ready for culling. Three udders were perfused with Tyrode's solution and three were not-perfused. During six hours, we collected perfusate samples for biochemical analysis. We also collected alveolar and teat canal tissue to evaluate gene expression. The biochemical parameters indicated that the perfused udders remained viable for the entire period of the experiment. A real-time polymerase chain reaction showed an increase in 18S rRNA gene expression in the alveolar tissue at 3 and 4 h after perfusion. There was also an increase in the Ubiquitin gene in the teat canal from not-perfused udders at 1, 3, and 4 h after slaughter. In general, gene expression was stable during the experiment. Our results indicated that the isolated perfused bovine udder model is appropriate for genetic studies, opening a new perspective in animal experimentation methods.

Association of exercise training and angiotensin-converting enzyme 2 activator improves baroreflex sensitivity of spontaneously hypertensive rats.

The present study sought to determine cardiovascular effects of aerobic training associated with diminazene aceturate (DIZE), an activator of the angiotensin converting enzyme 2, in spontaneously hypertensive rats (SHRs). Male SHRs (280-350 g) were either subjected to exercise training or not (sedentary group). The trained group was subjected to 8 weeks of aerobic training on a treadmill (five times a week, lasting 60 min at an intensity of 50-60% of maximum aerobic speed). In the last 15 days of the experimental protocol, these groups were redistributed into four groups: i) sedentary SHRs with daily treatment of 1 mg/kg DIZE (S+D1); ii) trained SHRs with daily treatment of 1 mg/kg DIZE (T+D1); iii) sedentary SHRs with daily treatment of vehicle (S+V); and iv) trained SHRs with daily treatment of vehicle (T+V). After treatment, SHRs were anesthetized and subjected to artery and femoral vein cannulation prior to the implantation of ECG electrode. After 24 h, mean arterial pressure (MAP) and heart rate (HR) were recorded; the baroreflex sensitivity and the effect of double autonomic blockade (DAB) were evaluated in non-anesthetized SHRs. DIZE treatment improved baroreflex sensitivity in the T+D1 group as compared with the T+V and S+D1 groups. The intrinsic heart rate (IHR) and MAP were reduced in T+D1 group as compared with T+V and S+D1 groups. Hence, we conclude that the association of exercise training with DIZE treatment improved baroreflex function and cardiovascular regulation.

Association of exercise training and angiotensin-converting enzyme 2 activator improves baroreflex sensitivity of spontaneously hypertensive rats

The present study sought to determine cardiovascular effects of aerobic training associated with diminazene aceturate (DIZE), an activator of the angiotensin converting enzyme 2, in spontaneously hypertensive rats (SHRs). Male SHRs (280–350 g) were either subjected to exercise training or not (sedentary group). The trained group was subjected to 8 weeks of aerobic training on a treadmill (five times a week, lasting 60 min at an intensity of 50–60% of maximum aerobic speed). In the last 15 days of the experimental protocol, these groups were redistributed into four groups: i) sedentary SHRs with daily treatment of 1 mg/kg DIZE (S+D1); ii) trained SHRs with daily treatment of 1 mg/kg DIZE (T+D1); iii) sedentary SHRs with daily treatment of vehicle (S+V); and iv) trained SHRs with daily treatment of vehicle (T+V). After treatment, SHRs were anesthetized and subjected to artery and femoral vein cannulation prior to the implantation of ECG electrode. After 24 h, mean arterial pressure (MAP) and heart rate (HR) were recorded; the baroreflex sensitivity and the effect of double autonomic blockade (DAB) were evaluated in non-anesthetized SHRs. DIZE treatment improved baroreflex sensitivity in the T+D1 group as compared with the T+V and S+D1 groups. The intrinsic heart rate (IHR) and MAP were reduced in T+D1 group as compared with T+V and S+D1 groups. Hence, we conclude that the association of exercise training with DIZE treatment improved baroreflex function and cardiovascular regulation.

Efeito da temperatura das amostras de leite na concentração de cálcio solúvel e de beta-caseína: interferência no teste de estabilidade frente ao etanol / Effect of milk sample temperature on the concentration of soluble calcium and beta-casein: interference with the ethanol stability test

O objetivo deste trabalho foi avaliar a variação do perfil proteico e do cálcio solúvel na coagulação do leite pelo etanol nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Amostras de leite de 61 animais foram avaliadas quanto à estabilidade ao etanol nas concentrações de 66 a 92 por cento (v/v) nas temperaturas de 4ºC, 10ºC, 15ºC e 20ºC. Três amostras, após 24 horas de armazenamento a 4ºC, foram ultracentrifugadas em quadruplicata (40.000 x g) a 4ºC e a 20ºC, respectivamente, por 60 minutos. Em seguida, o sobrenadante foi retirado e submetido à análise do cálcio solúvel pela técnica via úmida (digestão nitroperclórica) e leitura em espectrofotômetro de absorção atômica. O perfil proteico foi analisado pela técnica de eletroforese capilar empregando kit específico para determinação proteica. Os resultados mostraram uma correlação positiva entre o aumento da temperatura das amostras e a estabilidade do leite frente às diferentes concentrações de etanol. A porcentagem de cálcio solúvel no sobrenadante após ultracentrifugação foi maior nas amostras tratadas a 4ºC (P<0,05). As amostras ultracentrifugadas na temperatura de 4ºC apresentaram quantidades superiores de β-caseína no sobrenadante em comparação com as amostras tratadas a 20ºC. O abaixamento da temperatura favoreceu a migração da β-caseína e do cálcio coloidal para a fase solúvel do leite, o que possivelmente favoreceu o aumento da instabilidade das amostras no teste do etanol. Os resultados sugerem que a temperatura ideal para a realização de teste de estabilidade do leite frente ao etanol deveria ser de 21ºC...

The aim of this study was to evaluate the variation in protein profile and soluble calcium in milk coagulation by ethanol at 4ºC, 10ºC, 15ºC and 20ºC. Milk samples from 61 dairy cows were evaluated for stability of ethanol concentrations from 66 to 92 percent (v/v) at temperatures of 4°C, 10°C, 15°C and 20°C. Three samples were ultracentrifuged (40,000 x g) after 24 hours of storage at 4°C and 20°C, respectively, for 60 minutes. Their supernatants were removed and subjected to analyses of soluble calcium through nitro-perchloric digestion and atomic absorption spectrophotometry. The protein profiles were determined by capillary electrophoresis using a specific kit for protein determination. The results showed a positive correlation between the increase in temperature of the samples and the stability of milk against various concentrations of ethanol. The percentage of soluble calcium in the supernatant after centrifugation was higher in samples treated at 4°C (P<0.05). The samples ultracentrifuged at 4°C showed higher amounts of β-casein in the supernatant compared with samples stored at 20°C. The lowering of the temperature favored the migration of β-casein and colloidal calcium to the soluble phase of milk, which may also have favored the instability of milk in the ethanol test. According to the results, the milk sample temperature for the ethanol stability test should be 21ºC...

Incidence of bovine leukocyte adhesion deficiency, complex vertebral malformation, and deficiency of uridine-5-monophosphate synthase carriers in Brazilian Girolando cattle.

Among the various hereditary diseases that have been widely studied in dairy cattle, bovine leukocyte adhesion deficiency (BLAD), deficiency of uridine-5-monophosphate synthase (DUMPS), and complex vertebral malformation (CVM) are noteworthy because of their high impact on overall herd productivity as a consequence of increased calf mortality. The aim of this study was to verify the frequency of carriers of BLAD, CVM, and DUMPS mutant alleles in cows and bulls from the National Girolando Progeny Test carried out in Brazil by using polymerase chain reaction (PCR)-restriction fragment length polymorphism and allele-specific PCR assays. A total of 777 animals were genotyped for BLAD, 783 for CVM, and 122 for DUMPS. The frequencies of carriers for BLAD and CVM were 0.77 and 1.53%, respectively, whereas no carriers of DUMPS were observed.

Somatic development and embryo yield in crossbred F1 mice generated by different mating strategies.

The aim of this study was to evaluate different mating strategies among endogamic strains to create F1 populations of mice, minimising the effect of inbreeding depression on somatic development and embryo yield. Females from the strains Swiss, CBA and C57Bl/6 were divided in nine experimental mate arrangements. The total numbers of pups born alive per dam and somatic development, estimated by weighing and measuring the crown-rump length, were recorded. Superovulation response was evaluated in outbreed females. Litter size differed among endogamic dams, irrespective of the sire. Somatic development results suggest heterosis and imprinting phenomena, once a differential parental effect was demonstrated. There was no difference in corpora lutea, ova or embryos recovered (P > 0.05), but recovery and viability rates differ among F1 groups (P < 0.05). The association of dam prolificity with somatic development and superovulation response of the pups should be considered for experimental F1 populations establishment. The use of outbreed animals, however, did not reduce response variability to hormone treatment.