Identification of RSVP14 and RSVP20 components by two-dimensional electrophoresis and Western-blotting.

We have already shown that RSVP14 and RSVP20, two ram seminal plasma (SP) proteins postulated to be involved in sperm capacitation and gamete interaction can protect spermatozoa against cold-shock. In this study, we use two-dimensional polyacrylamide gel electrophoresis (2D-PAGE) for the analysis of SP proteins of Rasa Aragonesa rams, using enhanced protein solubilization in the presence of tributyl phosphine (TBP) and a polyacrylamide linear gradient gel with a narrow pH range (4-7). The image analysis of the 2D map detected 195 protein spots, with isoelectric points (pIs) ranging from 4.5 to 6.6, and molecular weight (M(r)) from 11.7 to 90.4. Staining of 2D gels with Pro-Q Emerald 300 Glycoprotein Stain revealed that most significant proteins in ram SP are glycosylated. The removing of protein N-linked oligosaccharides improved the gel resolution. 2D-PAGE analysis of the whole fraction 6 (F6) separated from ram SP by exclusion chromatography showed six main protein spots, four (a, b, c, d) in the 14 kDa and two (e, f) in the 20 kDa region. Western-blot analyses indicated that the anti-P14 antibody recognized four spots on the SP map, 4, 5, 6 and 7, that matched with spots a, b, c, d of F6 map. The anti-P20 antibody recognized spots 13 and 14 of SP map that corresponded to spots e, f of F6 map. The deduced sequences by de novo sequencing evidenced that protein spots 7 and 13 have significant similarities to BSP family, while protein spots 4 and 14 did not appear to be homologous with any reported protein in the current mammalian Proteinbank databases.     

Variation in seed protein digestion of different pea (Pisum sativum L.) genotypes by cecectomized broiler chickens: 1. Endogenous amino acid losses, true digestibility and in vitro hydrolysis of proteins

The aim of this study was to investigate the cause of variation in the digestibility of pea protein in poultry and to find a tool to select genotypes with high digestibility potential by using an in vitro hydrolysis assay. Eight pea genotypes were selected for their difference in seed protein content and composition. To reduce the variation due to tannins and particle size, seeds from these 8 genotypes were dehulled and micro-ground. They were incorporated as the only protein source in 8 different experimental isoproteinaceous diets with similar metabolisable energy content. The amino acid digestibility was studied in cecectomized chickens. A balance method was used to obtain apparent digestibility, and the isotope dilution technique was used to determine endogenous losses and true digestibility, after feeding a double labelled test meal containing chromic oxide and ¹⁵N-labelled peas. The 8 diets showed differences in apparent amino acid digestibility. The average apparent digestibility for all amino acids varied between 79.5 and 86.3%, with the highest values for arginine (85.2 to 90.8%) and glutamic acid (85.2 to 90.5%), and the lowest values for cystine (63.3 to 69.7%) and tryptophan (69.1 to 80.3%). This variability of apparent amino acid digestibility was due to variations in endogenous losses and true digestibility among the 8 pea genotypes. The average endogenous losses as determined for 9 amino acids ranged from 3.6 to 5.4% of ingested amino acids, with the highest value for threonine (8.0 to 11.0%). The average true digestibility varied between 84.4 and 90.2%, with the highest values for lysine (89.0 to 95.0%), and the lowest for isoleucine (81.0 to 88.7%) and valine (82.4 to 88.7%). In vitro hydrolysis of protein from micro-ground seeds was performed for the 8 pea genotypes using three proteases (pepsin, trypsin and chymotrypsin). The quantity of small peptides (< 3 kDa) that appeared after the combined hydrolysis with pepsin (3 h) followed by trypsin and chymotrypsin (15 min) was significantly correlated with the average true digestibility of the 8 genotypes (R = 0.74; P < 0.05).     

Risk factors associated with Mycoplasma capricolum subspecies capripneumoniae and morbillivirus infection in small ruminants in Tanzania

Tropical Animal Health and Production - Mortality of domestic small ruminants caused by contagious caprine pleuropneumonia (CCPP) and Peste des petits ruminants (PPR) is frequently reported in...     

The phenotypic relationship between residual intake and gain and other feed efficiency traits in Nellore cattle

This study aimed to compare feed efficiency measures of Nellore beef cattle on different residual intake and gain (RIG) classes. We used data from 610 animals weighing on average 236.33 kg and average of 283 days of age from feedlot performance tests carried out between 2005 and 2012. Animals were grouped based on RIG into three different classes: high RIG (>?mean?+?0.5 standard deviation (SD), most efficient; n?=?193), medium RIG (mean?±?0.5 SD; n?=?235), and low RIG (<?mean – 0.5 SD, least efficient; n?=?182). Residual feed intake (RFI), residual gain (RG), feed conversion ratio (FCR), feed efficiency (FE), relative growth rate (RGR), and Kleiber ratio (KR) of animals in each RIG class were compared by Tukey test at 1% of probability. Phenotypic correlations between variables were evaluated as well. Animals on high RIG class showed lower dry matter intake (P?<?0.01) and higher average daily gain (P?<?0.01) than low RIG animals. Consequently, high RIG animals had lower FCR (P?<?0.01) and higher FE (P?<?0.01) than those animals in low RIG class. The most efficient animals based on RIG were also the most efficient animals based on RG and RFI. RIG was negatively correlated to dry matter intake (P?<?0.01) and FCR (P?<?0.01), and a positive correlation was found between RIG and FE (P?<?0.01). Therefore, RIG appears to be a good parameter to select animals with reduced dry matter intake and high productive performance.

     

Reproductive behavior in hair sheep rams under tropical conditions

The reproductive efficiency of sheep herds depends to a great extent on the ram. Male reproductive evaluation allows to select for the best and eliminate those with reproductive problems. The objective was to evaluate the effect of breed and age group on the reproductive behavior of hair sheep rams in the tropics of Mexico. Pelibuey (n?=?42), Blackbelly (n?=?30), Dorper (n?=?44), and Katahdin (n?=?30) rams of two age groups: young (n?=?74, 1–1.5 years old) and adult (n?=?72, 2–4 years old) were evaluated. Serving capacity (10-min duration) and breeding soundness evaluation (BSE) tests were carried out in each ram. In the first test, rams were classified as suitable and unsuitable, and in the BSE test, they were classified as satisfactory, questionable, or unsatisfactory. The response variables were analyzed using chi-square test or analyses of variance (ANOVA). ANOVA included the fixed effects of breed, age group, and their interaction. In the serving capacity test, 79.5% of the rams were considered suitable, with no breed differences (P?>?0.05). Adult rams (90.3%) had the highest proportion of suitable rams (P?<?0.05). In the BSE test, 80.2% of the rams were satisfactory; only breed being significant (P?<?0.05). Pelibuey breed had the highest proportion of satisfactory rams (91.4%). Breed × age interaction was no significant for any trait. After serving capacity and BSE tests, a high proportion of rams was found not suitable for reproduction (36.3%), which is expected to cause low fertility in the flock.

     

Morphological and Immunohistochemical Characterization of Canine Osteosarcoma Spheroid Cell Cultures

Spheroid cell culture emerges as powerful in vitro tool for experimental tumour research. In this study, we established a scaffold‐free three‐dimensional spheroid system built from canine osteosarcoma (OS) cells (D17). Spheroids (7, 14 and 19 days of cultivation) and monolayer cultures (2 and 7 days of cultivation) were evaluated and compared on light and electron microscopy. Monolayer and spheroid cultures were tested for vimentin, cytokeratin, alkaline phosphatase, osteocalcin and collagen I by means of immunohistochemistry. The spheroid cell culture exhibited a distinct network of collagen I in particular after 19‐day cultivation, whereas in monolayer cultures, collagen I was arranged as a lamellar basal structure. Necrotic centres of large spheroids, as observed in 14‐ and 19‐day cultures, were characterized by significant amounts of osteocalcin. Proliferative activity as determined by Ki‐67 immunoreactivity showed an even distribution in two‐dimensional cultures. In spheroids, proliferation was predominating in the peripheral areas. Metastasis‐associated markers ezrin and S100A4 were shown to be continuously expressed in monolayer and spheroid cultures. We conclude that the scaffold‐free spheroid system from canine OS cells has the ability to mimic the architecture of the in vivo tumour, in particular cell–cell and cell–matrix interactions.     

Relation of lactose levels in milk and indicators of mammary gland health in the first third of lactation]

A total of 662 bucket milk samples from cows of two breed groups were examined: red and white breed = Bohemian Pied cattle with different genetic proportions of Ayrshire and Red Holstein improvement breeds; black and white breed = Black and White Lowland breed and different degree of absorptive crossing with a genetic proportion of the Holstein breed. Samples of daily milk yields were taken in the first three months of lactation once a month within a year. A possibility of using lactose content as an auxiliary indicator for detection of the mammary gland secretion disorders in the initial lactation stage was evaluated. The average values of the different indicators and their variability are summarized in Tab. I showing also the significance in a statistical model of included effects. Lactose content (L) was 4.88 +/- 0.20%, chloride content (Cl-) 113.7 +/- 22.4 mg/100 ml, somatic cell (SC) count 474 +/- 805 thousand/ml, SC count log corresponds to the geometrical mean of 234 thousand/ml, titratable acidity (SH) 7.34 +/- 0.83 x 2.5 mmol/l, chloride-lactose ratio (ClL) 2.27 +/- 0.51, conductivity (gamma) 442.4 +/- 34.5 mS/m and mastitis test (MT-NK) 0.72 +/- 1.18. The efficiency of the used statistical model was highest for Cl- content (Tab. I, R2 = 0.41), and it was lowest for SC counts (R2 = 0.07), while it increased to the twofold value (R2 = 0.15) after logarithmic transformation of SC counts. The breed group exerted a significant effect on Cl-, SC, log SC, SH, ClL, gamma and MT-NK (Tab. I). The breed group of red and white cows (Tab. II) had higher component contents and better indicators of the udder health state (Cl-, SC, log SC, ClL, gamma and MT-NK). The month of lactation influenced significantly SC, log SC, SH and gamma (Tab. I). A decrease in SC counts with the accruing month of lactation was observed (Tab. II), the trend of gamma and SH was opposite. The effect of lactation number was found to be significant for L, SC, log SC, SH, ClL, gamma and MT-NK (Tab. I). A tendency of a gradual decrease with the lactation number was observed in these indicators: L, SH, proteins and solids-non-fat (Fig. 1), while Cl-, gamma and ClL showed an opposite tendency. The year season influenced significantly L, log SC, SH, ClL, gamma and MT-NK (Tab. I, Fig. 2).(ABSTRACT TRUNCATED AT 400 WORDS)     

Antimicrobial susceptibility patterns of Haemophilus parasuis from pigs in the United Kingdom and Spain

A total of 30 British and 30 Spanish Haemophilus parasuis isolates were tested for their susceptibility to 19 of the antimicrobials currently used in swine practice with a broth microdilution method in order to know the emergence of resistance against these compounds in this porcine pathogen. All the British isolates were susceptible to penicillin, ceftiofur, erythromycin, tilmicosin, enrofloxacin, and florfenicol, and most of them were susceptible to the remaining antimicrobials (the highest resistance rate found was of 20% to neomycin). In contrast, all the Spanish isolates were susceptible exclusively to florfenicol, and high proportions of resistance were encountered for penicillin, ampicillin, oxytetracycline, erythromycin, tilmicosin, tiamulin and trimethoprim+sulphamethoxazole; in addition, a bimodal or multimodal distribution, or tailing of Spanish isolates over the MIC range was observed for clindamycin, sulphonamides and tylosine tartrate, suggesting the development of acquired resistance. In addition, several multiresistance patterns were found among the Spanish isolates, 23.3% of them being resistant to at least eight antimicrobials, the same rate as that encountered for those being susceptible to all antimicrobials tested. This study showed that in general British H. parasuis isolates are susceptible to antimicrobial agents routinely used for treatment of porcine respiratory diseases; however, the Spanish isolates need a more continuous surveillance of their susceptibility patterns.