Lippia gracilis Schauer essential oil as a growth promoter for Japanese quail.

The use of antibiotics as performance enhancers in animal feeding is declining, so Lippia gracilis Schauer essential oil (LGSEO) could be used as a potential substitute for the conventionally used growth promoters. The LGSEO contains components such as carvacrol and thymol, which kill and/or control pathogenic bacteria, increase population of beneficial organisms, act against oxidative processes and onto nutrient digestibility and absorption. The aim of this study was to investigate the action and the effects of LGSEO as a growth promoter in the diet of Japanese quail by examining their productive performance, intestinal microbiology, blood biochemical parameters, hepatic thiobarbituric acid reactive substances (TBARS) content and intestinal gene expression. A total of 252 two-day-old quail (Coturnix coturnix japonica) were assigned to 3 treatments in 7 replicates, using 12 birds per experimental unit. The treatments consisted of a basal diet, basal diet + LGSEO at 400 mg/kg of diet and basal diet + chemical antimicrobial (bacitracin methylene disalicylate) at 500 mg/kg of diet. The experimental period was 34 days. The highest feed intake (P < 0.01) was found in the group receiving the conventional antimicrobial, whereas the best feed conversion (P < 0.01) was shown by the animals receiving LGSEO. Escherichia coli growth was restricted in the quail receiving the growth promoters. Salmonella spp. growth was controlled by the treatment containing the conventional antimicrobial. There was no difference between the treatments (P > 0.05) for the concentration of aspartate aminotransferase and alanine aminotransferase enzymes in the blood or hepatic TBARS content. Birds receiving negative-control treatment exhibited a higher expression of sodium-glucose cotransporter (SGLT1), while those receiving the treatment with essential oil showed lower catalase (CAT) and glutathione peroxidase (GPX7) expressions compared to the conventional antimicrobial and control groups, respectively. Lippia gracilis Schauer essential oil is a powerful performance enhancer for Japanese quail by virtue of its abilities to improve their intestinal environment, balance the microbial population and reduce energy expenditure for oxidative processes.

Determinación de límites de referencia de analitos hematológicos en cabras lecheras adultas en el Altiplano Mexicano / Hematological analyte reference limits in adult dairy goats

RESUMEN Con el objetivo de establecer los límites de referencia de analitos hematológicos de cabras adultas de razas productoras de leche clínicamente sanas, sujetas al manejo y a las características de los sistemas de producción intensiva del Altiplano Mexicano, se recolectaron y analizaron muestras de sangre de cabras de 2 o más años de edad en diferentes etapas de producción. Con los resultados se obtuvieron los límites de referencia para hematocrito, cuenta de eritrocitos, volumen globular medio (VGM), plaquetas, sólidos totales, fibrinógeno, cuenta de leucocitos, neutrófilos segmentados, neutrófilos en banda, linfocitos, monocitos, eosinófilos y basófilos. Las medias obtenidas para cada analito fueron: hematocrito (0,22-0,36 L/L), cuenta de eritrocitos (9,07-18,47 x1012/L), volumen globular medio (VGM) (14,44-27,91), plaquetas (229-3 1 63 x109/L), solidos totales (65,64-84,64 g/L), fibrinógeno (0-6 g/L), cuenta de leucocitos (3,75-11,47 x109/L), neutrófilos segmentados (0,95-4,98 x109/L), neutrófilos en banda (0-0,16 x109/L), linfocitos (1,76-6,44 x109/L), monocitos (0-0,65 x109/L), eosinófilos (0-1,36 x109/L) y basófilos (0 - 0,37 x109/L). Estos valores se compararon con los obtenidos en otros estudios similares, y se encontraron algunas diferencias mayoritariamente en los valores obtenidos para hematocrito, eritrocitos, VGM, plaquetas, leucocitos y linfocitos, lo cual se debe a las diferencias en condiciones medioambientales y de alimientación entre los estudios.

ABSTRACT With the aim to establish the hematological analyte reference limits of clinically healthy adult goats of dairy producing breeds, subjected to the handling and characteristics of intensive production systems in the Mexican Plateau, the blood samples of 2-year old and older goats at different production stages were collected and analyzed. Reference limits (interval in brackets) for hematocrit (0.22-0.36 L/L), erythrocyte count (9.07-18.47 x1012/L), mean cell volume (MCV) (14.44-27.91 fL), platelets (229-3163 x109/L), total solids (65.64-84.64 g/L), fibrinogen (0-6 g/L), leukocyte count (3.75-11.47 x109/L), segmented neutrophils (0.95-4.98 x109/L), band neutrophils (0-0.16 x109/L), lymphocytes (1.76-6.44 x109/L), monocytes (0-0.65 x109/L), eosinophils (0-1.36 x109/L) and basophils (0-0.37 x109/L) were obtained. These values were compared with similar study results. Differences were found mainly in the values for hematocrit, erythrocytes, MCV, platelets, total leukocytes, and lymphocytes; this is due to environmental and feeding differences between the studies.

Antimicrobial activity of an aspartic protease from Salpichroa origanifolia fruits.

Plant proteases play a fundamental role in several processes like growth, development and in response to biotic and abiotic stress. In particular, aspartic proteases (AP) are expressed in different plant organs and have antimicrobial activity. Previously, we purified an AP from Salpichroa origanifolia fruits called salpichroin. The aim of this work was to determine the cytotoxic activity of this enzyme on selected plant and human pathogens. For this purpose, the growth of the selected pathogens was analysed after exposure to different concentrations of salpichroin. The results showed that the enzyme was capable of inhibiting Fusarium solani and Staphylococcus aureus in a dose-dependent manner. It was determined that 1·2 µmol l-1 of salpichroin was necessary to inhibit 50% of conidial germination, and the minimal bactericidal concentration was between 1·9 and 2·5 µmol l-1 . Using SYTOX Green dye we were able to demonstrate that salpichroin cause membrane permeabilization. Moreover, the enzyme treated with its specific inhibitor pepstatin A did not lose its antibacterial activity. This finding demonstrates that the cytotoxic activity of salpichroin is due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of the AP could represent a potential alternative for the control of pathogens that affect humans or crops of economic interest. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides insights into the antimicrobial activity of an aspartic protease isolated from Salpichroa origanifolia fruits on plant and human pathogens. The proteinase inhibited Fusarium solani and Staphylococcus aureus in a dose-dependent manner due to the alteration of the cell plasma membrane barrier but not due to its proteolytic activity. Antimicrobial activity of salpichroin suggests its potential applications as an important tool for the control of pathogenic micro-organisms affecting humans and crops of economic interest. Therefore, it would represent a new alternative to avoid the problems of environmental pollution and antimicrobial resistance.

Short communication: Effect of sampling time relative to the first daily feeding on interpretation of serum fatty acid and beta-hydroxybutyrate concentrations in dairy cattle.

Serum nonesterified fatty acid (NEFA) and beta-hydroxybutyrate (BHBA) concentrations are used to evaluate energy status in peripartum dairy cows. Blood samples from 37 cows in the week before parturition and 47 cows in the first week after parturition from 3 dairy herds were taken 1h before the first feeding (-1h) as well as 4 and 10h after the first feeding. Nonesterified fatty acid concentrations were measured in samples from cows before calving and BHBA was measured in samples from lactating cows. Mean NEFA concentrations in the prepartum cows were significantly higher at -1h (0.20 mmol/L) than at 4h (0.14 mmol/L), but were not different between 4 and 10h (0.17 mmol/L). Using a cutpoint of NEFA > or = 0.4 mmol/L, 32% of cows had high concentrations at -1h compared with 16% of the same cows at 4 and 10h. There were no differences in mean BHBA between -1h (646 micromol/L) and 4h (596 micromol/L), but mean BHBA was higher at 10h (711 micromol/L) than at -1h. Using a cutpoint of BHBA > or = 1,400 micromol/L, there were no differences in the proportions of high BHBA, which were 9, 11, and 13% of cows at -1, 4, and 10h, respectively. Prandial effects on serum NEFA may affect interpretation of this analyte. In order not to misclassify cows when assessing energy status, samples for NEFA must at least be taken at a consistent time relative to feeding within a given herd. When sampling cows to monitor elements of energy metabolism in the prepartum period, there was twice the probability of detecting animals with NEFA values > or = 0.4 mmol/L if they were sampled 1h before the first feed delivery compared with sampling the same cows 4 or 10h after feeding.

Comparative analysis of six different antibodies against Her2 including the novel rabbit monoclonal antibody (SP3) and chromogenic in situ hybridisation in breast carcinomas.

AIMS: To compare the sensitivity and specificity of new rabbit monoclonal antibody SP3 with those of mouse monoclonal and rabbit polyclonal antibodies using HER2 amplification defined by chromogenic in situ hybridisation (CISH) as the gold standard. METHODS: Serial sections from tissue microarrays (TMAs) containing 84 breast carcinomas were submitted to CISH (Zymed HER2 Spot-Light kit) and immunohistochemistry, using NeoMarkers SP3 (rabbit monoclonal), DAKO A0485 and DAKO HercepTest (polyclonal), Novocastra NCL-CB11, Cell Marque CM-CB11, and Genentech 4D5 (mouse monoclonal). RESULTS: The best antibody concordance was between SP3 and HercepTest (kappa = 0.74). SP3, A0485 and HercepTest detected all HER2 amplified tumours, but were less specific than mouse monoclonal antibodies. 3/38 (7.9%) and 8/38 (21.0%) non-amplified tumours were scored as 3+ using SP3 and A0485, respectively. 3/46 (6.5%) amplified tumours were negative for NCL-CB11. SP3, HercepTest and A0485 showed no gene amplification on 55%, 62.5% and 92.3% of the 2+ scored tumours, but most of the 2+ scored tumours using monoclonal antibodies were amplified by CISH (80-92.3%). CONCLUSIONS: SP3 is more sensitive than mouse monoclonal antibodies for Her2 assessment. However, HercepTest, CB11 and 4D5 show higher specificity than SP3 for the identification of HER2 gene amplification. Mouse monoclonal antibodies show less Her2 2+ tumours; most are amplified by CISH.

Blood acid-base and serum electrolyte values in red deer (Cervus elaphus).

Acid-base, serum electrolyte, plasma protein, and packed cell volume (PCV) values were determined in venous blood samples from 30 red deer (Cervus elaphus) of both sexes showing no clinical signs of disease. The animals were 5 months of age and kept on pasture in the Valley of Mexico, at an altitude of 2450 m. Blood samples were collected without sedation. Mean blood values were: pH 7.411 +/- 0.041, pCO2 37.7 +/- 4.4 mmHg, base excess 0.7 +/- 3.2 mmol/L, actual bicarbonate 24.3 +/- 3.1 mmol/L, total CO2 25.3 +/- 3.2 mmol/L and anion gap 23.5 +/- 5.5 mmol/L. Mean serum electrolyte levels were: Na+ 142.3 +/- 2.5 mmol/L, Cl- 100.5 +/- 2.3 mmol/L, and K+ 7.03 +/- 1.03 mmol/L. Plasma protein and PCV values were 60.0 +/- 6.6 g/L and 0.47 +/- 0.05 L/L, respectively. Blood values determined in this study can be considered reference data for health control and disease diagnosis.