Binucleate Rhizoctonia sp. AG G causing root rot in yacon (Smallanthus sonchifolius) in Brazil

A new rot caused by a binucleate Rhizoctonia sp. affecting the tuberous root cortex of the domesticated yacon ( Smallanthus sonchifolius ) has been observed in Brazil. Isolates of a binucleate Rhizoctonia sp. were collected from roots with rot symptoms and characterized by the number of nuclei per cell, hyphal anastomosis, RAPD molecular markers, ITS-5·8S rDNA sequence and pathogenicity tests. All isolates had a mean of 1·9–2·2 nuclei per cell and anastomosed with the binucleate Rhizoctonia sp. AG G-tester strain. RAPD analysis was carried out between 11 isolates recovered from yacon and 11 AG (A, Ba, Bb, Bo, C, D, F, G, O, P, Q) standard testers of binucleate Rhizoctonia sp. Genetic similarities of 94·8–100% were observed among isolates of the binucleate Rhizoctonia sp. from yacon and all isolates were genetically more closely related to the AG G tester than other strains according to upgma analysis using RAPD markers. Homologies of complete ITS nucleotide sequences were 100% between binucleate isolates of Rhizoctonia sp. from yacon and the AG G tester. According to pathogenicity tests, the isolates caused typical rot symptoms of yacon tubers 90 days after inoculation     

Interleukin-8 as a prognostic serum marker in canine mammary gland neoplasias

Mammary gland tumors in female dogs are an excellent model for the clinic-pathological, diagnostic and prognostic investigation of mammary neoplasias. Prognostic and predictive markers are effective in research and routine diagnosis. Interleukins play a fundamental role in cancer, with a particular function in tumor growth, invasion and metastatic potential. Interleukin-8 (IL-8) is known to possess tumorigenic and pro-angiogenic properties, and its overexpression is seen in a number of human tumors. IL-8 serum levels were determined and correlated with the clinic-pathological features and clinical evolution of mammary gland neoplasias in female dogs. IL-8 was measured by an immunoenzymatic assay in 30 female dogs with mammary neoplasias within a 12 month follow-up and in 50 control animals. The correlation between IL-8 concentration and clinical parameters was investigated. A statistically significant difference in the IL-8 serum levels was found in tumor-bearing dogs compared to the controls. In addition, when the individual parameters were evaluated, IL-8 content showed a positive correlation with the tumor progression, lymph node involvement, recurrence and death. Single and multivariate analyses showed associations between tumor recurrence, metastasis, high clinical staging and high IL-8, and also with the death risk. This was also consistent with the high IL-8 content in dogs showing tumor recurrence and metastasis. IL-8 superexpression has been detected in a number of human tumors, usually associated with a poor prognostic. Besides promoting angiogenesis, IL-8 is strongly related with the metastatic phenotype of mammary tumor cells. High IL-8 concentration was found in mammary gland cancer patients with advanced disease stages. Our results show that IL-8 can be used as a non-invasive prognostic marker for mammary gland cancer, and can be useful for the prediction of disease progression and recurrence in dogs with mammary neoplasias. The increased level of this cytokine acts as an independent prognostic marker of survival and the identification of animals with the poor prognostic.     

Validation of an ELISA method for the serological diagnosis of canine brucellosis due to Brucella canis

In the present study, the validation of an enzyme-linked immunosorbent assay (ELISA) for serodiagnosis of canine brucellosis is described. Two different antigenic extracts, obtained by heat or ultrasonic homogenization of microbial antigens from a wild isolate of Brucella canis bacteria, were compared by ELISA and Western blot (WB). A total of 145 canine sera were used to define sensitivity, specificity and accuracy of the ELISA as follows: (1) sera from 34 animals with natural B. canis infection, confirmed by blood culture and PCR, as well as 51 sera samples from healthy dogs with negative results by the agar–gel immunodiffusion (AGID) test for canine brucellosis, were used as the control panel for B. canis infection; and (2) to scrutinize the possibility of cross reactions with other common dog infections in the same geographical area in Brazil, 60 sera samples from dogs harboring known infections by Leptospira sp., Ehrlichia canis, canine distemper virus (CDV), Neospora caninum, Babesia canis and Leishmania chagasi (10 in each group) were included in the study. The ELISA using heat soluble bacterial extract (HE-antigen) as antigen showed the best values of sensitivity (91.18%), specificity (100%) and accuracy (96.47%). In the WB analyses, the HE-antigen showed no cross-reactivity with sera from dogs with different infections, while the B. canis sonicate had various protein bands identified by those sera. The performance of the ELISA standardized with the heat soluble B. canis antigen indicates that this assay can be used as a reliable and practical method to confirm infection by this microorganism, as well as a tool for seroepidemiological studies.     

Effects of different types of dietary non-digestible carbohydrates on the physico-chemical properties and microbiota of proximal colon digesta of growing pigs

The effect of different types of fibre on the intestinal digesta and microbial diversity was evaluated in growing pigs. The animals were fed during forty-two days with one of four experimental diets including a control group, a diet enriched with resistant starch type I, by coarse ground corn, and other two diets containing sugar beet pulp (8%) and wheat bran (10%) (as sources of soluble and insoluble non-starch polysaccharides respectively). Body weight (BW) and average daily feed intake (ADFI) were assessed at day 7, 21 and 42. Luminal samples of digesta from proximal colon were collected to analyse water retention capacity (WRC), concentration of short-chain fatty acids (SCFA) and the microbial diversity. Overall, animals fed with higher amounts of non-digestible carbohydrates had lower ADFI and BW. Sugar beet pulp provoked an increase of WRC, a higher concentration of SCFA, and a more stable microbial diversity throughout the experimental period. On the other hand, animals fed with wheat bran presented a lower bacterial diversity.     

Effects of limited concentrate feeding on growth and blood and serum variables, and on nutrient digestibility and gene expression of hepatic gluconeogenic enzymes in dairy calves

This study elucidated the effects of limited concentrate feeding on growth, nutrient digestibility, blood profile and gene expression of gluconeogenic enzymes in the liver of dairy calves. The study utilized 36 German Holstein dairy calves (5-7 days of age) divided into two groups of 18 calves each for 150 days. Control group calves received 2 kg/(calf × day) of concentrate, whereas calves in the restricted group received only 1 kg/(calf × day). Good quality forage (mixture of maize and grass silages) was available for ad libitum consumption to both groups. The intake of milk replacer before weaning, and of concentrate were recorded daily per calf; however, the consumption of forages was quantified as daily average of the group. Body weights (BW) were recorded at start and on days 35, 70, 112 and 150. Blood and serum samples and spot urinary and faecal samples were also collected at similar time points. On days 70 and 150, liver biopsies were collected from seven animals in each group. The BW was not different between the groups at all times. Total BW gain in the control group was 124 kg as opposed to 111 kg in restricted group that led to average BW gain of 827 g/day and 739 g/day in respective groups, and the differences were significant (p = 0.018). As planned, the control group had higher concentrate and lower forage intake than the restricted group. The blood haemoglobin, haematocrit and serum variables (glucose, total protein, albumin and urea) were within the normal range in both groups, but serum glucose was higher (p < 0.05) in control than in restricted group at 70 days. There was no difference between groups in organic matter (OM) digestibility which declined (p < 0.001) with increasing age in both groups. Microbial crude protein (MCP) synthesis estimated from urinary allantoin excretion increased (p < 0.001) in both groups with increasing age but was not different between groups. The mRNA expressions for the gluconeogenic enzymes, cytosolic and mitochondrial phosphoenol pyruvate carboxykinase (EC 4.1.1.32) and pyruvate carboxylase (EC 6.4.1.1) measured by quantitative real-time PCR in liver biopsies showed no differences between groups. Overall, restricting concentrate moderately reduced the growth intensity without affecting the normal serum and blood indices, and MCP synthesis and OM digestibility showed no differences between groups, indicating that both concentrate feeding schemes can be successfully applied.     

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.     

Fish functional diversity responses following flood pulses in the upper Paraná River floodplain

Flood pulses are the main force driving the dynamics of aquatic communities in floodplains. The responses of communities to environmental changes following flood pulses usually demand a time lag to appear and reach the climax. We assembled a data set of 16 years of fish samplings to assess the relationship between water level and four functional diversity measures, in the upper Paraná River floodplain. Specifically, we approached four aspects of each relationship between water level and functional diversity: nature (positive or negative), sensitivity (response intensity), responsiveness (response delay) and extent (response duration). The nature of the relationship between water level and functional diversity was positive in all cases. Functional richness (FRic) responded right after flood pulses, although with shorter extent. Abundance‐dependent functional measures (evenness—FEve; divergence—FDiv; and Rao's quadratic entropy—Rao's Q) presented delayed responses, reaching peaks more than 1.5 years after flood pulses. Significant effects of floods on fish functional diversity were observed for more than 3 years, although the highest functional diversity was observed with 1.8 years, on average. More importantly, flood pulses had no longer significant effects on functional diversity after 4 years. Regarding conservation strategies in regulated systems, flood events should occur every 2 or 3 years, with adequate timing (October‐November), intensity (up to 450 cm) and duration (at least 50 uninterrupted days). Intervals longer than 3 years or inadequate timing, intensity and duration could dramatically decrease functional diversity and compromise ecosystem services.     

Triploid Induction in the Yellowtail Tetra,Astyanax altiparanae,Using Temperature Shock: Tools for Conservation and Aquaculture

Triploidization is an interesting tool to produce sterile fish. In the yellowtail tetra, Astyanax altiparanae, this can be applied for aquaculture and surrogate technologies. In this study, we compared the efficacy of cold (2 C) or heat shock (38 C, 40 C, and 42 C) on triploid induction in the yellowtail tetra. The eggs were treated with cold or heat shock, 2 min postfertilization (30 min in cold shock or 2 min in heat shock). Intact embryos served as the control group. Ploidy status was confirmed by karyotyping, flow cytometry, and nuclear diameter of erythrocytes. The hatching rate decreased after cold shock (12.69 ± 15.76%) and heat shock at 42 C (0.35 ± 0.69%) in comparison with the control group (63.19 ± 16.82%). At 38 C and 40 C, hatching rates (61.29 ± 17.73% and 61.75 ± 22.1%, respectively) were not decreased. Only one triploid arose at 38 C (1/80). At 40 C, a high number of triploids arose (72/78). At 42 C, very few embryos developed into the hatching stage. A large number of haploid individuals arose after cold shock (61/75), with only one triploid. Our results indicate that heat shocking of embryos at 40 C is optimum for triploid production in the yellowtail tetra.