Using BCG,MPT-51 and Ag85 as antigens in an indirect ELISA for the diagnosis of bovine tuberculosis

This study evaluated the Mycobacterium tuberculosis protein antigen MPT-51, the trimeric antigen 85 (Ag85) complex, and Bacillus Calmette-Guérin (BCG) in an indirect ELISA to diagnose bovine tuberculosis (TB) from serum samples. Serum was collected from 208 intra-dermal tuberculin test (ITT)-positive and 54 ITT-negative animals from a region where bovine TB is endemic. Using the Ag85 and BCG antigens, the indirect ELISA was able to discriminate ITT-positive from ITT-negative animals. This level of discrimination was not achieved when using the MPT-51 antigen. The highest sensitivity (Se) and specificity (Sp) of the test was found when BCG was used (Se, 82%; Sp, 91%). Further work in different epidemiological settings and with larger numbers of animals will be required to validate these findings.     

Duodenal perforation caused by Rhizomucor species in a cat

CLINICAL SUMMARY: A 7-month-old female Persian cat presented with gastrointestinal (GI) necrosis and perforation caused by Rhizomucor species. Unfortunately, the cat died of bacterial peritonitis and sepsis before a definitive diagnosis, based on histopathology and fungal culture, was achieved. PRACTICAL RELEVANCE: This appears to be the first reported case of GI disease caused by Rhizomucor species in a cat. Mucorales infections typically cause acute and rapidly progressive disease. As illustrated by this case, clinicians should be alert to the potentially fatal consequences of an opportunistic Rhizomucor species infection in their feline patients.     

Iron Sources Effects on Growth,Physiological Parameters and Nutrition of Cacao

Productivity and sustainability of cacao (Theobroma cacao L.) in tropical soils are affected by levels of iron. Information is lacking on the cacao response to various sources of iron (Fe). A greenhouse experiment was conducted to evaluate the effects of five iron sources iron sulfate heptahydrate, ferric ethylenediamine-N,N’-bis(2-hydroxyphenylacetic acid), ferric diethylenetriaminepentaacetic acid, ferric ethylenediaminetetraacetic acid, fiesta herbicide (FeSO₄ · 7H₂O, FeEDDHA, FeDTPA, FeEDTA,) at 10 mg Fe kg^?1 soil on growth, photosynthesis, content of photosynthetic pigments and starch and macro- and micronutrient nutrition of cacao. The various iron sources had significant effects on shoot and root dry biomass accumulation, leaf chlorophyll a and b content, carotenoid levels, SPAD index and P_N. These parameters were significantly correlated with concentration, uptake, influx, and transport and use efficiency of Fe. In cacao net photosynthesis, stomatal conductance, internal carbon dioxide (CO₂)_, and transpiration in leaf level responded differently to the sources of Fe. Invariably, macro and micronutrient uptake, influx, transport, and use efficiency showed differential responses to sources of iron but significant effects were only observed for copper (Cu), Fe, manganese (Mn), and zinc (Zn). Overall, FeDTPA, FeEDTA and FeHEDTA could be the best sources of Fe in improving, growth, photosynthesis and macro and micro nutrition of cacao.     

In vivo bioinsecticidal activity toward Ceratitis capitata (fruit fly) and Callosobruchus maculatus (cowpea weevil) and in vitro bioinsecticidal activity toward different orders of insect pests of a trypsin inhibitor purified from tamarind tree (Tamarindus indica) seeds

A proteinaceous inhibitor with high activity against trypsin-like serine proteinases was purified from seeds of the tamarind tree (Tamarindus indica) by gel filtration on Shephacryl S-200 followed by a reverse-phase HPLC Vidac C18 TP. The inhibitor, called the tamarind trypsin inhibitor (TTI), showed a Mr of 21.42 kDa by mass spectrometry analysis. TTI was a noncompetitive inhibitor with a Ki value of 1.7 x 10(-9) M. In vitro bioinsecticidal activity against insect digestive enzymes from different orders showed that TTI had remarkable activity against enzymes from coleopteran, Anthonomus grandis (29.6%), Zabrotes subfasciatus (51.6%), Callosobruchus maculatus (86.7%), Rhyzopertha dominica(88.2%), and lepidopteron, Plodia interpuncptella (26.7%), Alabama argillacea (53.8%), and Spodoptera frugiperda (75.5%). Also, digestive enzymes from Diptera, Ceratitis capitata (fruit fly), were inhibited (52.9%). In vivo bioinsecticidal assays toward C. capitata and C. maculatus larvae were developed. The concentration of TTI (w/w) in the artificial seed necessary to cause 50% mortality (LD50) of larvae was 3.6%, and that to reduce mass larvae by 50.0% (ED50) was 3.2%. Furthermore, the mass C. capitata larvae were affected at 53.2% and produced approximately 34% mortality at a level of 4.0% (w/w) of TTI incorporated in artificial diets.     

Redistribution of 10B Absorbed by Leaves and Roots of Cashew Seedlings

Boron (B) is transported mainly in the xylem, but by using isotopic tracers it was possible to verify that B has significant mobility in the phloem of polyols-producing species. Thus, the objective of this study was to evaluate the mobility of ¹⁰B applied to cashew plants. The experiment was conducted from 05/10 to 07/10/2010 in a greenhouse of University of São Paulo State-UNESP, Ilha Solteira Campus, State of São Paulo, Brazil. 0.5 mg of B L^?1 of soil was used in the substrate. A solution of 255 mg of B L^?1 was used in recently mature and fully developed leaves. The total content of B and and percentage of boron in the plant derived from fertilizer (Bppf%) were evaluated in the old and new leaves stemming at 30 and 60 days after the application of ¹⁰B. As evidenced, B applied to leaves had restricted mobility in cashew tree. However, B applied in the substrate was absorbed and translocated to young leaves.     

Integrating geospatial and multi‐depth laboratory spectral data for mapping soil classes in a geologically complex area in southeastern Brazil

Soil mapping across large areas can be enhanced by integrating different methods and data sources. This study merges laboratory, field and remote sensing data to create digital maps of soil suborders based on the Brazilian Soil Classification System, with and without additional textural classification, in an area of 13 000 ha in the state of São Paulo, southeastern Brazil. Data from 289 visited soil profiles were used in multinomial logistic regression to predict soil suborders from geospatial data (geology, topography, emissivity and vegetation index) and visible–near infrared (400–2500 nm) reflectance of soil samples collected at three depths (0–20, 40–60 and 80–100 cm). The derived maps were validated with 47 external observations, and compared with two conventional soil maps at scales of 1:100 000 and 1:20 000. Soil suborders with and without textural classification were predicted correctly for 44 and 52% of the soil profiles, respectively. The derived suborder maps agreed with the 1:100 000 and 1:20 000 conventional maps in 20 and 23% (with textural classification) and 41 and 46% (without textural classification) of the area, respectively. Soils that were well defined along relief gradients (Latosols and Argisols) were predicted with up to 91% agreement, whereas soils in complex areas (Cambisols and Neosols) were poorly predicted. Adding textural classification to suborders considerably degraded classification accuracy; thus modelling at the suborder level alone is recommended. Stream density and laboratory soil reflectance improved all classification models, showing their potential to aid digital soil mapping in complex tropical environments.     

The dispersal of Culicoides brevitarsis in eastern New South Wales and associations with the occurrences of arbovirus infections in cattle

Distributions of the vector Culicoides brevitarsis Kieffer (Diptera: Ceratopogonidae) (determined from light trap data) and 2 arboviruses (determined from seroconversions in sentinel cattle) were studied in eastern New South Wales in 1993–94. C brevitarsis was recorded progressively from endemic areas on the north coast, to Nowra on the south coast, and westward to Scone, in the Hunter Valley. C brevitarsis also survived through winter at Paterson, in the Hunter Valley. Its apparently focal reappearance in this marginal area had no obvious effect on the broad pattern of its progression or the dispersal of Akabane and bluetongue viruses. These viruses were first recorded from foci near Coffs Harbour, on the mid-north coast. Their first occurrences at different locations were associated with those of C brevitarsis, but not with each other. The viruses were found only within the recorded limits of the vector's distribution. Delays between the initial occurrence of C brevitarsis and first evidence of virus transmissions at locations ranged from 2 to 7 months. The delays decreased away from the points of focus and were negatively associated with the time of initial occurrence of the vector. Seroconversions to the viruses were related to the presence of C brevitarsis. However, the densities of C brevitarsis had no apparent effect on the initial numbers of cattle seroconverting to either virus. The results support the conclusion that the progressions of C brevitarsis and Akabane and bluetongue viruses were the result of gradual movements by the vector.     

Estimated BVDV-prevalence, -contact and -vaccine use in dairy herds in Northern Portugal

A study to evaluate BVDV-prevalence, recent -contact and -vaccine use in dairy herds in the "Entre Douro e Minho" (EDM) region in North Portugal was carried out in 124 dairy herds in 2003. Herds were visited to ascertain BVDV-vaccine use and to collect a bulk tank milk (BTM) sample and serum from 1268 cattle to analyse BVDV-antibodies using an NS2-3 ELISA. Fifty-three percent of farmers used inactivated BVDV-vaccines whilst the remaining farmers were not presently using BVDV-vaccines. BMT-antibody results included 35% positives, 25% negative and 39% inconclusive, and were similar in vaccinated and non-vaccinated herds (p>0.05) and allowed estimating a 10% BVDV herd-prevalence from prior knowledge of the relationship between BMT-antibody results and probability of PI cattle in the herd. Overall individual seroprevalence was 27% and was 23% in non-vaccinated and 36% in vaccinated cattle (p<0.05). Contact of the herd with BVDV was assessed according to seroprevalence in young and adult cattle in the herd and it was estimated that 35% of herds were infected or had recent contact with BVDV, 40% were not infected and did not have recent contact with BVDV and the BVDV-infection and -contact status of remaining herds was undetermined. The results from this study indicate BVDV is endemic and BVDV-vaccines are widespread in the dairy-cattle population in EDM region in Portugal.     

Mitochondrial damage as an early event of monensin-induced cell injury in cultured fibroblasts L929

The present study was designed to identify, submicroscopically, the primary organelle or target structure for monensin in cultured murine fibroblasts L929. In addition, the effect of the drug on cell size and surface membranes of the cells were analysed; cellular proliferation, collagen secretion, and necrosis and apoptosis were re-evaluated. At the lowest concentration of monensin the foremost ultrastructural alteration occurred in the mitochondria, characterized by increased matrix density with disorganized and less distinct crystae. Incubation with monensin at higher concentrations resulted in severe mitochondrial damage and marked dilatation of the Golgi apparatus and rough endoplasmic reticulum cisternae. Fibroblasts exposed to higher concentrations of monensin were enlarged with decreased number of filopodia and hollows in the surface membrane. Moreover, monensin inhibited the cell proliferation, increased immunohistochemical positiveness for collagen type I in a dose-dependent manner, and, at high concentrations, caused cell necrosis whereas apoptosis was not induced. Taken together, these results show that monensin induces early mitochondrial damage, possibly causing an energy deficit that led to inhibition of fibroblasts proliferation and accumulation of collagen causing dilatation of Golgi apparatus and rough endoplasmic reticulum. Moreover, the mitochondrial damage would also explain the monensin-induced necrosis.     

Serotypes and virulence markers of Shiga toxin-producing Escherichia coli (STEC) isolated from dairy cattle in São Paulo State, Brazil

In order to determine the occurrence, serotypes and virulence markers of Shiga toxin-producing Escherichia coli (STEC) strains, 153 fecal samples of cattle randomly selected from six dairy farms in Sao Paulo State, Brazil, were examined for Shiga toxin (Stx) production by the Vero cell assay. Feces were directly streaked onto MacConkey Sorbitol Agar and incubated at 37 degrees C overnight. Sorbitol-negative colonies (maximum 20) and up to 10 sorbitol-positive colonies from each plate were subcultured onto presumptive diagnostic medium IAL. Sorbitol-negative isolates were screened with O157 antiserum for identification of O157:H7 E. coli. Isolates presenting cytotoxic activity were submitted to colony hybridization assays with specific DNA probes for stx1, stx2, eae, Ehly and astA genes. The isolation rate of STEC ranged from 3.8 to 84.6% depending on the farm analysed. STEC was identified in 25.5% of the animals, and most of them (64.1%) carried a single STEC serotype. A total of 202 STEC isolates were recovered from the animals, and except for the 2 O157:H7 isolates all the others expressed cytotoxic activity. The great majority of the STEC isolates carried both stx1 and stx2 genes (114/202, 56.4%) or stx2 (82/202, 40.6%); and whereas the Ehly sequence occurred in most of them (88%) eae was only observed in O157:H7 and O111:HNM isolates. Serotypes O113:H21, O178:H19 and O79:H14 were the most frequent STEC serotypes identified and widely distributed among animals from different farms, while others such as O77:H18, O88:H25 and O98:H17 occurred only in particular farms. This is the first report on the occurrence of STEC in dairy cattle in Sao Paulo State, and the results point to substantial differences in rate of isolation, serotypes and genetic profile of STEC that has been previously described among beef cattle in our community. Moreover, to our knowledge O79:H14 and O98:H17 represent new STEC serotypes, while O178:H19 has only been recently reported in Spain.