Comparative digestibility of low‐quality grass hay by two breeds of cattle differing in mature live weight

The digestive capacity of the Portuguese native breed of Barrosão cattle and the Holstein–Friesian breed was measured when fed meadow hay (72–74 g CP and 641–671 g NDF/kg dry matter), offered either alone or supplemented with soya bean meal (150 g/kg dry matter), at maintenance level. Four mature cows of each breed were used. Average initial live weight (LW) was 457 and 635 kg for the Barrosão and the Holstein–Friesian cows respectively. The organic matter digestibility (OMD) of the meadow hay was higher in Holstein–Friesian than in Barrosão cows (p < 0.08) whether supplemented or not. The neutral detergent fibre digestibility of the hay was also higher in Holstein–Frisian when the hay was fed alone (p < 0.08). The soya bean meal supplementation increased the OMD and the NDFD digestibility of the total diet (p < 0.05), but not the OMD digestibility of the hay (p > 0.05). The results obtained in this study suggest a higher ability to digest fibre in the large dairy breed than in the small native breed. This suggestion is also supported by previous findings with sheep breeds largely differing in mature live weight.     

Content of plasmatic glycerol and activity of hepatic glycerol kinase in broiler chickens fed diets containing different sources and concentrations of glycerine

The objective of this study was to evaluate the effect of three sources of glycerine (crude glycerine from soya bean oil – CGSO, mixed crude glycerine from frying oil and lard – MCG, and a semipurified glycerine from soya bean oil – SPGSO) in four concentrations in the diet (17.5, 35.0, 52.5 and 70.0 g of each type of glycerine per kg of feed) on the levels of plasmatic glycerol and the activity of the hepatic enzyme glycerol kinase in broilers of 22–35 days old (experiment I) and 33–42 days old (experiment II). The highest (p < 0.05) plasmatic glycerol level was detected in broilers fed diet containing CGSO. Independent of the source, increasing the concentration of glycerine led to a linear increase (p < 0.05) in the plasmatic glycerol concentration. In experiment I, all the diets containing glycerine resulted in increased concentrations of plasmatic glycerol, in relation to the control diet without glycerine. However, in experiment II, only the diet containing 17.5 g of CGSO per kg of feed and the diets formulated with any of the three types of glycerine in the inclusion concentrations of 35, 52.5 and 70 g/kg of feed resulted in higher plasmatic glycerol levels than those observed in the control broilers. The source of glycerine influenced the glycerol kinase activity only in experiment II, where the use of CGSO in the diet increased the enzyme activity. For both experiments, the glycerol kinase activity increased with the inclusion of glycerine in the diet. In conclusion, based on the absence of saturation of the glycerol kinase activity for the three glycerine sources and for both rearing periods evaluated, the broilers can metabolize the glycerol (at the level of the phosphorylation of the glycerol to glycerol‐3‐phosphate in the liver) present in the glycerine when the diet is supplemented with up to 70 g of glycerine per kg of feed.     

Gonadotropin‐induced Puberty Does Not Impair Reproductive Performance of Gilts over Three Parities

The aim of this study was to evaluate the reproductive performance of three parities of gilts treated or not treated with gonadotropin to induce puberty. Sixty gilts received 600 IU of equine chorionic gonadotropin (eCG) followed by 2.5 mg of porcine luteinizing hormone (LH) 72 h later. Fifty‐nine other gilts were exposed only to a mature boar for 15 min twice daily. Artificial insemination (AI) was performed at 0, 12 and 24 h after the detection of oestrus, and gestation was confirmed by ultrasound after 35 days. Sows were inseminated at the first post‐weaning oestrus. The total numbers of piglets born, piglets born alive, stillborn, mummified foetuses, as well as pregnancy and farrowing rates were evaluated for each of the three parities. Culling rates, farrowing intervals and weaning‐to‐oestrous intervals (WEI) were also analysed. Mean age at puberty and oestrous manifestation were not significantly different between treatments (p = 0.0639; 179.20 ± 17.52 compared with 173.96 ± 16.94, 91.66% compared with 94.92%) across the experimental period. However, females that underwent puberty induction showed modest increases both in the number of total pigs born and in the number of piglets born alive. In conclusion, puberty induction through exogenous gonadotropin administration in field conditions did not induce a more concentrated first oestrous manifestation, but trended to a modest increase in the number of pigs born alive in the first parity and a reduced culling rate during the first gestation.     

Perkinsus sp. infestation in carpet-shell clams, Ruditapes decussatus (L), along the Portuguese coast. Results from a 2-year survey

This study characterises Perkinsus atlanticus disease prevalence and intensity within the economically important beds of the clam Ruditapes decussatus along the Iberian Atlantic coast. Samples were collected from five different sites along the Portuguese coast (Ria Formosa, Ria do Alvor, V.N. Milfontes, Aveiro, Lagoa de Obidos) and in Galicia (Ria de Arosa) between winter 2000/2001 and 2002/2003. The infection level was evaluated by the Ray Fluid Thioglycollate medium (RFTM) method using the body burden assay. In addition, the measure of the condition index (CI) (percentage between the edible part and the total weight of the clams) was conducted to investigate a possible correlation between this parameter and the intensity of the infection. However, no clear relationship between infection intensity and condition index (CI) could be determined although a significant decline in CI was found for heavily infected clams. Results of a 2-year survey demonstrate the presence of Perkinsus sp. in all sampling sites albeit with different intensities. Sites where human interference was minimal showed the lowest levels of infection. There were no significant differences in Perkinsus sp. infection intensity between samples collected in winter and summer, in contrast with several studies describing higher intensities at the end of the summer. Nevertheless, major differences were observed from year to year and site to site, indicating that factors other than those responsible for seasonal climatic variations might affect the prevalence and the intensity of Perkinsus sp. infection.     

Lipid-based transfection as a method for gene delivery in zebrafish (Danio rerio) embryos

A major challenge to the widespread production of transgenic, knockout and knockdown zebrafish has been the absence of a simple and effective procedure for introducing macromolecules into the fertilized egg. None of the existing techniques for gene transfer in fish embryos has proven to be a major advance over cytoplasm microinjection, which is a technically demanding and time‐consuming procedure. This report addresses this need, considering that the development of protocols for lipid‐based transfection with fish embryos would considerably simplify gene transfer in this complex biological model. In this study, lipid‐based transfection with two different reporter vectors was carried out in zebrafish embryos at different developmental stages. The parameters tested included different plasmid/transfection reagent ratios as well as the influence of an added transfection enhancer reagent. When embryos were transfected in the blastula stage with a pEGFP‐N1 vector, more than 35% successfully incorporated the plasmid and expressed the fluorescent protein 24 h after transfection. The transfection enhancer did not show any significant effect in our experiments. This work presents an approach to implement this technique as a faster, cheaper and more practical alternative than microinjection.     

Antioxidant and Antimicrobial Potential of the Bifurcaria bifurcata Epiphytic Bacteria

Surface-associated marine bacteria are an interesting source of new secondary metabolites. The aim of this study was the isolation and identification of epiphytic bacteria from the marine brown alga, Bifurcaria bifurcata, and the evaluation of the antioxidant and antimicrobial activity of bacteria extracts. The identification of epiphytic bacteria was determined by 16S rRNA gene sequencing. Bacteria extracts were obtained with methanol and dichloromethane (1:1) extraction. The antioxidant activity of extracts was performed by quantification of total phenolic content (TPC), 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity and oxygen radical absorbance capacity (ORAC). Antimicrobial activities were evaluated against Escherichia coli, Pseudomonas aeruginosa, Bacillus subtilis, Salmonella enteritidis, Staphylococcus aureus, Saccharomyces cerevisiae and Candida albicans. A total of 39 Bifurcaria bifurcata-associated bacteria were isolated and 33 were identified as Vibrio sp. (48.72%), Alteromonas sp. (12.82%), Shewanella sp. (12.26%), Serratia sp. (2.56%), Citricoccus sp. (2.56%), Cellulophaga sp. (2.56%), Ruegeria sp. (2.56%) and Staphylococcus sp. (2.56%). Six (15.38%) of the 39 bacteria Bifurcaria bifurcata-associated bacteria presented less than a 90% Basic Local Alignment Search Tool (BLAST) match, and some of those could be new. The highest antioxidant activity and antimicrobial activity (against B. subtilis) was exhibited by strain 16 (Shewanella sp.). Several strains also presented high antimicrobial activity against S. aureus, mainly belonging to Alteromonas sp. and Vibrio sp. There were no positive results against fungi and Gram-negative bacteria. Bifurcaria bifurcata epiphytic bacteria were revealed to be excellent sources of natural antioxidant and antimicrobial compounds.     

Maritime Halophyte Species from Southern Portugal as Sources of Bioactive Molecules

Extracts of five halophytes from southern Portugal (Arthrocnemum macrostachyum, Mesembryanthemum edule, Juncus acutus, Plantago coronopus and Halimione portulacoides), were studied for antioxidant, anti-inflammatory and in vitro antitumor properties. The most active extracts towards the 1,1-diphenyl-2-picrylhydrazyl (DPPH) radical were the methanol extracts of M. edule (IC₅₀ = 0.1 mg/mL) and J. acutus (IC₅₀ = 0.4 mg/mL), and the ether extracts of J. acutus (IC₅₀ = 0.2 mg/mL) and A. macrostachyum (IC₅₀ = 0.3 mg/mL). The highest radical scavenging activity (RSA) against the 2,2′-azino-bis (3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) radical was obtained in the ether extract of J. acutus (IC₅₀ = 0.4 mg/mL) and H. portulacoides (IC₅₀ = 0.9 mg/mL). The maximum total phenolic content (TPC) was found in the methanol extract of M. edule (147 mg gallic acid equivalents (GAE)/g) and in the ether extract of J. acutus (94 mg GAE/g). Significant decreases in nitric oxide (NO) production were observed after incubation of macrophages with lipopolysaccharide (LPS) and the chloroform extract of H. portulacoides (IC₅₀ = 109 µg/mL) and the hexane extract of P. coronopus (IC₅₀ = 98.0 µg/mL). High in vitro cytotoxic activity and selectivity was obtained with the ether extract of J. acutus. Juncunol was identified as the active compound and for the first time was shown to display selective in vitro cytotoxicity towards various human cancer cells.     

Host/parasite relationship in the in vitro infection of rat gliocytes by Neospora caninum: evaluation of cell respiration

Neospora caninum is a protozoon that causes abortion in cattle and neuromuscular lesions in dogs, with the formation of cysts mainly in the central nervous system. Since N. caninum is an intracellular parasite with tropism for the cells of nervous system, this study evaluated the respiratory metabolism of glial cells infected by this* parasite. Glial cultures obtained from the cerebral cortex of newborn rats were kept in DMEM enriched with 10% fetal bovine serum, 1 mM pyruvic acid and 2 mM of L-glutamine. They were infected at a ratio of approximately 1:1 (cell/parasite). Oxygen consumption was evaluated by polarography in the non infected and N. caninum infected groups, 24 and 72 h following infection. Glial cell respiration after 24 and 72 h was 307.2 +/- 34.7 and 308.9 +/- 64.1 microL of oxygen per mug of total protein per minute, and 566.2 +/- 54.6 and 579 +/- 117.5 microL O2/microg of total protein/minute in the control and infected groups, respectively. These results show that N. caninum does not interfere with glial respiration in vitro.     

Resistance to common bacterial blight in Phaseolus vulgaris L. recombinant inbred lines under natural infection of Xanthomonas axonopodis pv. phaseoli

Among the main causes of poor yield in common beans are fungal, viral and bacterial diseases. Common bacterial blight, caused by Xanthomonas axonopodis pv. phaseoli (Xap), is one of the major bacterial diseases leading to significant losses in Brazil. Chemical control is ineffective, therefore, the use of resistant varieties becomes an interesting alternative. The objective of the present work was to evaluate disease resistance under natural infection of the pathogen in 109 recombinant inbred lines (F₇) of P. vulgaris originated from the cross HAB-52 (susceptible — snapbean) × BAC-6 (resistant — common bean) in two different environments, as well as to calculate genetic parameters to assist in the selection of promising materials to be used in the CBB resistance breeding program. The data of the genetic parameters were compared to those calculated for the F₃ generation originated from the same cross. The heritability results for DI (disease index) and VI (variation index) in F₃ were 26.85% and 0.26, respectively, whereas in F₇ they were 91.77% and 1.36, respectively. These results demonstrate a potential to be explored for this advanced population, that in the future, along with other pathogen variability studies and tests in other environments, may provide more information regarding a more precise evaluation of promising genotypes to be used in common bean breeding programs aiming to obtain CBB resistant varieties. This revised version was published online in July 2006 with corrections to the Cover Date.     

Crops use-efficiency of nitrogen from manures permitted in organic farming

The current increase in the organic agriculture segment has created a new market for fertilisers permitted for use in organic farming. Off-farm N sources for organic farming are scarce, considering the restriction on the use of chemical fertilisers. Thus, when some products are permitted in organic agriculture, commercial opportunities become available. In this study we compare the performances of Vegethumus (Veg) and Phenix (Phe), two manures that are permitted in organic farming, with several other manures, ammonium nitrate (AN) and control treatments. A 3-year field trial and a pot experiment were carried out in order to estimate dry matter yield, N uptake, and N nutritional status of the crops, as well as soil N availability, the latter was assessed by using anion exchange membranes inserted into the soil. Apparent N recovery (ANR) values in the field trial were 6.3% and 58.2% in Veg and AN plots, respectively, after the application of 380 kg N/ha in the previous five growing seasons. In the pot experiment, the ANR of Veg and Phe, the organic amendments permitted in organic farming, were 5.0% and 13.6%, while Beiraadubo (Bei) and Nutrisoil (Nut) had ANR of 27.2% and 42.0%. The poor results of the amendments permitted in organic farming, in light of their high prices suggest that their use must be carefully considered by farmer in their fertilization strategies.