Comparing p20’s RNA silencing suppressing activity among five phylogenetic groups of <Emphasis Type="Italic">Citrus Tristeza virus</Emphasis>

The p20 protein encoded by the Citrus Tristeza Virus (CTV) was previously identified as a RNA silencing suppressor. In this study, we analyzed the p20’s suppressing activity from five phylogenetic groups of CTV, using the co-infiltration assay of Green fluorescence protein (GFP) gene and the suppressor gene in 16C line Nicotiana benthamiana plants. Green fluorescence, GFP mRNA relative levels and GFP specific siRNAS were compared showing in most cases, only slight differences. Contrary to previous studies, the p20 suppressor was not able to impede neither short range nor systemic spreading of RNA silencing. The suppressor from the phylogenetic group 4 revealed a much reduced activity when compared with the others. At present we still don’t know whether this property is a characteristic of this group or an atypical feature due to a unique point mutation. The differences in the symptom type and intensity originated by isolates belonging to the phylogenetic groups assayed could not be related to differences to the p20 suppressor’s activity.     

Clinical evaluation of cochlear hearing status in dogs using evoked otoacoustic emissions

Objectives : Evoked otoacoustic emission testing is the preferred test in human patients for sensorineural deafness screening in neonates and cochlear outer hair cell function monitoring in adults. This study evaluated evoked otoacoustic emission testing for cochlear function assessment in dogs within a clinical setting. Methods : Two populations of anaesthetised dogs were included. In group 1 the evoked otoacoustic emission response was compared to the brainstem auditory evoked response in 10 dogs having hearing assessment. Group 2 comprised 43 presumed normal dogs, in which the suitability of two types of evoked otoacoustic emissions, transient‐evoked and distortion product otoacoustic emissions, were evaluated (brainstem auditory evoked response was not performed in this group). Results : Valid transient‐evoked otoacoustic emission and distortion‐product otoacoustic emission responses were successfully recorded within the clinical setting and correctly identified deaf and hearing ears. Within presumed healthy dogs, normal otoacoustic emission response was demonstrated in more than 80% of dogs using a single, short distortion‐product otoacoustic emission run and in 78% of dogs with valid transient‐evoked otoacoustic emission responses using a series of three repeated transient‐evoked otoacoustic emission short runs. Clinical Significance : Transient‐evoked otoacoustic emission and distortion‐product otoacoustic emission testing provided a rapid, non‐invasive frequency‐specific assessment of cochlear function. Transient‐evoked otoacoustic emission and distortion product otoacoustic emission testing is suitable as a screening procedure to detect loss of cochlear function in dogs, although further investigation is needed.     

Evaluation of Soy Protein Concentrate and Meat and Bone Meal as a Replacement for Fish Meal in the Diet of Nile Tilapia Fingerlings

This experiment evaluated soy protein concentrate (SPC) and meat and bone meal as a replacement for fish meal protein (FP) in the diet of Nile tilapia fingerlings. Five diets were evaluated including a control diet (50.0% crude protein [CP], of which half the protein came from fish meal [FM]), and four diets in which FM was replaced by increasing amounts of SPC (25.5, 50.0, 75.5, and 100.0%) in combination with meat and bone meal. At the conclusion of a 60‐d growth trial, the following parameters were determined: final weight (FW), final length (FL), weight gain (WG), feed intake (FI), apparent feed conversion (AFC), survival (SUR), and production cost. The estimate of the best replacement value was determined by polynomial regression and the linear response plateau. The results showed significant (P < 0.05) improvement for WG and AFC ratio as FM was gradually replaced by SPC. The parameters FL and FI reached their optimum value at a replacement level of 100% SPC. These results show that it is feasible to use a combination of SPC and meat and bone meal in a balanced feed formulation as an alternative protein source in the diets of Nile tilapia fingerlings.     

Detection of IgM antibodies against Babesia bovis in cattle

The diagnosis of acute babesiosis by direct examination of blood smears has some limitations and the indirect serological methods currently in use are designed for detection of IgG, which may not be detectable at an early stage of infection. There is a need, therefore, for rapid and reliable procedures to diagnose acute infections. An ELISA system using a crude antigenic preparation of Babesia bovis was standardized for the detection of IgM antibodies. Optimal dilutions of the antigen, using positive and negative reference sera, were determined by checkerboard titrations. Serum samples of cattle imported from tick-free areas collected before and during an immunization process were used to validate the tests. The specificity was 94% and sensitivity 100%. Specific IgM antibodies against B. bovis first appeared on the 11th day post-inoculation (p.i.) in animals infested with Boophilus microplus ticks and on the 19th day p.i. in animals which had been inoculated with infected blood. Antibody titers decreased after Day 33; however, all animals remained positive until the end of the experiment (124 days).     

Canine visceral leishmaniasis in São José de Ribamar, Maranhão State, Brazil

Here, we describe the situation of canine visceral leishmaniasis in two villages of S?o José de Ribamar in Maranh?o State/Brazil, where human cases have been registered. Blood samples of 36 household crossbred dogs from Sergio Tamer village and 43 dogs from Quinta village were collected and the serum used for serological diagnosis. An Indirect Fluorescent Antibody Test (IFAT) and enzyme-linked immunosorbent assay (ELISA) were used to detect antibodies against Leishmania. The clinical examination showed that 25% of the canine population of Quinta presented a poor body condition and in 39%, ectoparasites (ticks and fleas) were detected. In both tests, serology revealed that 21% (9 out of 43) of the dogs presented antibodies against Leishmania (55% were asymptomatic and 45% were symptomatic). In the Vila Sérgio Tamer, 25% (9 out of 36) of the dogs were seropositive for Leishmania (66.67% were asymptomatic and 33.33% were symptomatic), 33% presented poor body condition, and 22% have ectoparasites. The clinical signs more frequent were skin lesions. The statistical analysis showed that there was no statistical difference (p>0.05) between the seropositivity of the dogs from the two villages. The same was observed when the clinical signs were compared (p>0.05). Both villages have favorable conditions to maintain the cycle of leishmaniasis.     

Linear,logarithmic, and polynomial models of M-mode echocardiographic measurements in dogs

OBJECTIVE: To determine whether logarithmic and polynomial models are superior to simple linear models for predicting reference values for M-mode echocardiographic variables in dogs with a wide range of body weights. ANIMALS: 69 apparently healthy adult male and female dogs of various breeds, ages (range, 1 to 12 years; median, 3.5 years), and body weights (range, 3.9 to 977 kg; median, 25.4 kg). PROCEDURE: Echocardiographic M-mode measurements of the interventricular septum, left ventricular dimension (LVD), left ventricular wall, aorta, and left atrium were obtained. Simple linear, second-order polynomial, third-order polynomial, and logarithmic regression models were determined by use of the least-squares method to describe the relationship between M-mode measurements and body weight. Differences in adjusted R2 values of logarithmic and polynomial models were tested for significance of contribution, compared with the simple linear model. RESULTS: Significant differences in adjusted R2 were found when comparing simple linear with logarithmic or polynomial models for LVD-diastole, LVD-systole, aorta, and left atrium. Differences in adjusted R2 between second-order polynomial, third-order polynomial, and logarithmic models were not significant for any M-mode measurement. CONCLUSIONS AND CLINICAL RELEVANCE: In this study, logarithmic or second-order polynomial models predicted reference values of M-mode measurements for size of the cardiac chambers better than simple linear models for dogs with a wide range of body weights. Logarithmic and polynomial models were not superior to simple linear models for M-mode measurements of cardiac wall thickness.     

Oviductal Fluid Proteins Associated with the Bovine Zona Pellucida and the Effect on In Vitro Sperm–Egg Binding,Fertilization and Embryo Development

Studies have demonstrated that oviductal fluid (ODF) proteins associate with eggs of numerous species including the bovine. In this study, the association of three ODF proteins, the bovine oestrus‐associated protein, osteopontin (OPN), lipocalin‐type prostaglandin D synthase (L‐PGDS), with the bovine zona pellucida (ZP) was demonstrated by immunohistochemistry and western blot. The biological function of ODF derived egg‐associated OPN and L‐PGDS in sperm binding, fertilization and embryonic development was also explored. In vitro matured bovine oocytes were pre‐incubated with ODF collected by cannula from cows in oestrus, or ODF with antibodies to OPN, L‐PGDS and bovine serum albumin (BSA). Following incubation, oocytes were inseminated with 1 × 10⁵ frozen‐thawed spermatozoa, and they were evaluated for sperm binding, fertilization and embryonic development in vitro. Pre‐treatment of ODF with antibodies to all of proteins reduced sperm binding to the ZP and fertilization in vitro. Cleavage rates were not significantly different among incubations, but rates of embryo development were significantly decreased. We conclude that antibodies to OPN, L‐PGDS and BSA react with oocytes incubated with ODF and inhibit sperm binding, fertilization and embryonic development in vitro, suggesting a potential role of these proteins in these events.     

Comparative analysis of heart functions in micropigs and conventional pigs using echocardiography and radiography

The production of miniature animals has been suggested for use in organ transplantation. At present, many of the studies about application of animal organs to human have been focused on pigs because of the number of advantages involved and due to their similarities with human. However, a physiological analysis of the organs to be transplanted has not yet been carried out. Therefore, this study analyzed whether or not there were physiological and morphological differences in the hearts of conventionally-reared pigs and micropigs. In this study, the morphological and physiological functions of the heart were examined using radiographic and echocardiographic equipment. In the lateral radiographic view, the heart of the micropig has a larger cardiac long axis : short axis ratio than does the conventional pig, but the difference in the vertebral heart score was not significant. In addition, there were no morphological differences on the X-ray fluoroscopic view. There were no differences in echocardiographic values, except for several values in the left ventricle traces. Overall, it is expected that the values measured in this study will contribute to understanding of the physiological characteristics of micropigs.     

Biogeochemical cycles of nutrients in tropical Eucalyptus plantations: Main features shown by intensive monitoring in Congo and Brazil

The sustainability of fast-growing tropical Eucalyptus plantations is of concern in a context of rising fertilizer costs, since large amounts of nutrients are removed with biomass every 6–7 years from highly weathered soils. A better understanding of the dynamics of tree requirements is required to match fertilization regimes to the availability of each nutrient in the soil. The nutrition of Eucalyptus plantations has been intensively investigated and many studies have focused on specific fluxes in the biogeochemical cycles of nutrients. However, studies dealing with complete cycles are scarce for the Tropics. The objective of this paper was to compare these cycles for Eucalyptus plantations in Congo and Brazil, with contrasting climates, soil properties, and management practices.The main features were similar in the two situations. Most nutrient fluxes were driven by crown establishment the two first years after planting and total biomass production thereafter. These forests were characterized by huge nutrient requirements: 155, 10, 52, 55 and 23 kg ha^?1 of N, P, K, Ca and Mg the first year after planting at the Brazilian study site, respectively. High growth rates the first months after planting were essential to take advantage of the large amounts of nutrients released into the soil solutions by organic matter mineralization after harvesting. This study highlighted the predominant role of biological and biochemical cycles over the geochemical cycle of nutrients in tropical Eucalyptus plantations and indicated the prime importance of carefully managing organic matter in these soils. Limited nutrient losses through deep drainage after clear-cutting in the sandy soils of the two study sites showed the remarkable efficiency of Eucalyptus trees in keeping limited nutrient pools within the ecosystem, even after major disturbances. Nutrient input–output budgets suggested that Eucalyptus plantations take advantage of soil fertility inherited from previous land uses and that long-term sustainability will require an increase in the inputs of certain nutrients.