Cryptosporidium and Giardia in wild otters (Lutra lutra)

A total of 437 faecal samples from wild otter (Lutra lutra) were collected from 161 sites in Galicia (northwest Spain) and were analysed by a direct immunofluorescence antibody test (IFAT). Cryptosporidium sp. oocysts and Giardia sp. cysts were detected in 17 (3.9%) and 30 (6.8%) samples, respectively. The results demonstrate that otters may contribute to the contamination of watercourses, although further studies are required to establish which species or genotypes of Cryptosporidium and Giardia infect these animals and also the significance in terms of public health.     

Presence of Shiga toxin-producing E. coli O157:H7 in a survey of wild artiodactyls

This study was carried out to evaluate the role of wild artiodactyls as reservoirs of Escherichia coli O157:H7 for livestock and humans. Retroanal mucosal swabs samples from 206 red deer (Cervus elaphus), 20 roe deer (Capreolus capreolus), 6 fallow deer (Dama dama) and 11 mouflon (Ovis musimon), collected during the hunting season (autumn-winter) in South-western Spain, were screened. Samples were pre-enriched in modified buffered peptone water, concentrated by an immunomagnetic separation technique and cultured onto selective cefixime tellurite sorbitol MacConkey agar. Polymerase chain reaction (PCR) was used to detect the presence of genes coding O157 and H7 antigens and the virulence factors verocytotoxin, intimin and enterohaemolysin. Three E. coli O157:H7 isolates were obtained from red deer (1.5%). Two of them showed inability to ferment sorbitol and lack of beta-d-glucuronidase (GUD) activity, however, the other strain investigated was an atypical sorbitol-fermenting E. coli O157:H7 with GUD(+) activity. This is the first report pointing to red deer as a reservoir of E. coli O157:H7 in Spain.     

Feasibility of diode-array instruments to carry near-infrared spectroscopy from laboratory to feed process control

Near-infrared calibrations were developed for the instantaneous prediction of the chemical and ingredient composition of intact compound feeds. Two rather different instruments were compared (diode array vs grating monochromator). The grating monochromator was used in a static mode in the laboratory, whereas the diode-array instrumentbetter adapted to online analysiswas placed on a conveyor belt to simulate measurements at a feed mill plant. Modified partial least squares (MPLS) equations were developed using the same set of samples analyzed in the two instruments. Sample set 1 ( N = 398) was used to predict crude protein (CP) and crude fiber (CF), while sample set 2 ( N = 393) was used for the prediction of one macroingredient (sunflower meal, SFM) and one microingredient (mineral-vitamin premix, MVP). The standard error of cross-validation (SECV) and the coefficient of determination (R2) values for CF were better using the monochromator instrument. However, results obtained for CP, SFM, and MVP using the samples analyzed in the diode-array instrument showed similar or even greater accuracy than those obtained using samples analyzed in the grating monochromator. The excellent predictive ability [R2> 0.95; RPD (ratio of standard deviation to SECV) > 3] obtained for CP, CF, and SFM opens the way for the online use of NIRS diode-array instruments for surveillance and monitoring in the manufacture, processing, and marketing of compound feeds. R2, RPD, and SECV values for MVP showed similar performance for both instruments. Although RPD values did not reach the minimum recommended for quantitative analysis, results are encouraging for an ingredient present in feed compounds in such very low amounts.     

beta-Conglycinins among sources of bioactives in hydrolysates of different soybean varieties that inhibit leukemia cells in vitro

Soybean is a complex matrix containing several potentially bioactive components. The objective was to develop a statistical model to predict the in vitro anticancer potential of soybean varieties based on the correlation between protein composition and bioactive components after simulated gastrointestinal enzyme digestion with their effect on leukemia mouse cells. The IC 50 values of the hydrolysates of soy genotypes (NB1-NB7) on L1210 leukemia cells ranged from 3.5 to 6.2 mg/mL. Depending on genotype, each gram of soy hydrolysates contained 2.7-6.6 micromol of total daidzein, 3.0-4.7 micromol of total genistein, 0.5-1.3 micromol of glycitein, 2.1-2.8 micromol of total saponins, 0.1-0.2 micromol of lunasin, and 0.1-0.6 micromol of Bowman-Birk inhibitor (BBI). The IC 50 values calculated from a partial least-squares (PLS) analysis model correlated well with experimental data ( R (2) = 0.99). Isoflavones and beta-conglycinin positively contributed to the cytotoxicity of soy on L1210 leukemia cells. Lunasin and BBI were potent L1210 cell inhibitors (IC 50 = 13.9 and 22.5 microM, respectively), but made modest contributions to the activity of defatted soy flour hydrolysates due to their relatively low concentrations. In conclusion, the data demonstrated that beta-conglycinins are among the major protein components that inhibit leukemia cell growth in vitro. Furthermore, it was feasible to differentiate soybean varieties on the basis of the biological effect of their components using a statistical model and a cell-based assay.     

Taking NIR calibrations of feed compounds from the laboratory to the process: calibration transfer between predispersive and postdispersive instruments

In the context of current demands in the animal feed industry for controls and analyses, the use of instruments that may be applied on the process line has acquired a significant interest. A key aspect is that the calibrations developed for quality control with instruments sited in the laboratory (at-line) must be transferred to instruments that will be used in the plant itself (online). This study evaluates the standardization and the calibration transfer between a grating monochromator instrument (predispersive) designed for laboratory analysis and a diode array instrument (postdispersive) more adapted to process conditions. Two procedures that correct differences between spectra of two instruments were tested: the patented algorithm by Shenk and Westerhaus and piecewise direct standardization (PDS). Although results were slightly better with PDS, both methods achieved good spectral matching between the two instruments, with levels of repeatability similar to that of the grating instrument itself. The calibration transfer was evaluated in terms of the standard error of prediction (SEP), which was considerably reduced after standardization. However, final calibration models to be used in the diode array instrument must contain spectra from both types of instruments to give acceptable prediction accuracy.     

Identification and characterization of topoisomerase II inhibitory peptides from soy protein hydrolysates

Topoisomerases are targets of several anticancer agents because their inhibition impedes the processes of cell proliferation and differentiation in carcinogenesis. With very limited information available on the inhibitory activities of peptides derived from dietary proteins, the objectives of this study were to employ co-immunoprecipitation to identify inhibitory peptides in soy protein hydrolysates in a single step and to investigate their molecular interactions with topoisomerase II. For this, soy protein isolates were subjected to simulated gastrointestinal digestion with pepsin and pancreatin, and the human topoisomerase II inhibitory peptides were co-immunoprecipitated and identified on a CapLC- Micromass Q-TOF Ultima API system. The inhibitory activity of these peptides from soy isolates toward topoisomerase II was confirmed using three synthetic peptides, FEITPEKNPQ, IETWNPNNKP,and VFDGEL, which have IC 50 values of 2.4, 4.0, and 7.9 mM, respectively. The molecular interactions of these peptides evaluated by molecular docking revealed interaction energies with the topoisomerase II C-terminal domain (CTD) (-186 to -398 kcal/mol) that were smaller than for the ATPase domain (-169 to -357 kcal/mol) and that correlated well with our experimental IC 50 values ( R (2) = 0.99). In conclusion, three peptides released from in vitro gastrointestinal enzyme digestion of soy proteins inhibited human topoisomerase II activity through binding to the active site of the CTD domain.     

Anti-Inflammatory,Antioxidant, and Antimicrobial Effects of Underutilized Fish Protein Hydrolysate

Antimicrobial, anti-inflammatory, and antioxidant activities of protein hydrolysates from Argentine croaker (Umbrina canosai) protein isolate (CPI) or Argentine croaker myofibrillar protein with different degrees of hydrolysis (DH: 10–20%) prepared using Alcalase or Protamex were determined. Results showed that an increase in the DH resulted in higher content of hydrophobic and aromatic amino acids (AAAs) and in a decrease in molecular weight (MW) distribution for all hydrolysates obtained. Furthermore, the enzyme and raw material used influenced the amino acid content and MW determined. Hydrolysates from CPI with a 20% DH by Alcalase had higher 2,2?-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) radical scavenging activity, metal chelation, and ferric-reducing antioxidant power (p < 0.05). All hydrolysate samples decreased the pro-inflammatory capacity. In all the evaluated microorganisms, only seven were inhibited, most being Gram-positive. Alcalase was found to exert a considerable influence on antibacterial activity. These hydrolysates are an alternative as natural antimicrobials, anti-inflammatory, and antioxidant compounds.     

Genetic diversity of <Emphasis Type="Italic">Patellifolia patellaris</Emphasis> from the Iberian Peninsula,a crop wild relative of cultivated beets

The genetic diversity of Patellifolia patellaris has been investigated to generate information required for the organisation of a systematic genetic resources conservation action combining the best elements of the ex situ and in situ conservation concept. To this end, ten occurrences of the species were sampled on the Iberian Peninsula in Portugal and Spain. A total of 271 individuals was analysed using 24 microsatellite markers. The factorial analysis separated the material into two occurrences from Portugal, two occurrences sampled east of Gibraltar and six occurrences from the eastern Spanish coast. The pairwise genetic distance between occurrences and the complementary compositional differentiation among occurrences were calculated. The genetic distance values were used to construct an Unweighted Pair Group Method with Arithmetic Mean tree, which can be separated into four equidistant clusters. Two clusters are located in Portugal and two further clusters in Spain. The factorial analysis as well as the genetic distance and differentiation analysis indicate that the two occurrences from Portugal are clearly genetically different from the Spanish group of occurrences. Both occurrences have low population sizes and are therefore highly endangered. In situ conservation actions are urgently required for both occurrences. Further investigations are needed to organise better informed conservation actions for P. patellaris, namely to study genetic relationships between occurrences on the Spanish mainland and occurrences distributed on the Canary Islands, Madeira and Cape Verde Islands as well as in Morocco.