Antigenic and genetic characterization of twenty-six strains of human respiratory syncytial virus (subgroup A) isolated during three consecutive outbreaks in Havana city, Cuba.

Twenty-six human respiratory syncytial virus strains (subgroup A) isolated from three outbreaks in Havana City during the period 1994/95, 1995/96 and 1996/97 were analyzed to determine their antigenic and genetic relationships. Analyses were performed by monoclonal antibodies and restriction mapping (N gene) following amplification of the select region of the virus genome by polymerase chain reaction. All isolated strains were classified as subgroup A by monoclonal antibodies and they showed a restriction pattern NP4 that belonged to subgroup A. Thus the results obtained in this work, showed a close relation (100%) between antigenic and genetic characterization of the isolated strains in our laboratory. These methods permit the examination of large numbers of isolates by molecular techniques, simplifying the researchs into the molecular epidemiology of the virus.     

Conversion of an inactive cardiac dihydropyridine receptor II-III loop segment into forms that activate skeletal ryanodine receptors

A 25 amino acid segment (Glu666-Pro691) of the II-III loop of the alpha1 subunit of the skeletal dihydropyridine receptor, but not the corresponding cardiac segment (Asp788-Pro814), activates skeletal ryanodine receptors. To identify the structural domains responsible for activation of skeletal ryanodine receptors, we systematically replaced amino acids of the cardiac II-III loop with their skeletal counterparts. A cluster of five basic residues of the skeletal II-III loop (681RKRRK685) was indispensable for activation of skeletal ryanodine receptors. In the cardiac segment, a negatively charged residue (Glu804) appears to diminish the electrostatic potential created by this basic cluster. In addition, Glu800 in the group of negatively charged residues 798EEEEE802 of the cardiac II-III loop may serve to prevent the binding of the activation domain.     

Antilisterial Activity of Peptide AS-48 and Study of Changes Induced in the Cell Envelope Properties of an AS-48-Adapted Strain of Listeria monocytogenes

The peptide AS-48 is highly active on all Listeria species. It has a bactericidal and bacteriolytic mode of action on Listeria monocytogenes CECT 4032, causing depletion of the membrane electrical potential and pH gradient. The producer strain Enterococcus faecalis A-48-32, releases sufficient amounts of AS-48 into the growth medium to suppress L. monocytogenes in cocultures at enterococcus-to-listeria ratios above 1 at 37°C or above 10 at 15°C. As the temperature decreases, the bactericidal effects of AS-48 are less pronounced, but at 2.5 μg/ml it still can inhibit the growth of listeria at 6°C. AS-48 is highly active on liquid cultures, although concentrations above 0.2 μg/ml are required to avoid adaptation of listeria. AS-48-adapted cells can be selected at low (but still inhibitory) concentrations, and they can be inhibited completely by AS-48 at 0.5 μg/ml. The adaptation is lost gradually upon repeated subcultivation. AS48^ad cells are cross-resistant to nisin and show an increased resistance to muramidases. Their fatty acid composition is modified: they show a much higher proportion of branched fatty acids as well as a higher C_{15:0 An}-to-C_{17:0 An} ratio. Resistance to AS-48 is also maintained by protoplasts from AS48^ad cells. Electron microscopy observations show that the cell wall of AS48^ad cells is thicker and less dense. The structure of wild-type cells is severely modified after AS-48 treatment: the cell wall and the cytoplasmic membrane are disorganized, and the cytoplasmic content is lost. Intracytoplasmic membrane vesicles are also observed when the wild-type strain is treated with high AS-48 concentrations.     

Mytilid mussels: global habitat engineers in coastal sediments

Dense beds of mussels of the family Mytilidae occur worldwide on soft-bottoms in cold and warm temperate coastal waters and have usually been considered hot spots of biodiversity. We examined intertidal mussel beds at four distant locations around the globe with the same sampling method, to find out whether this “hot spot” designation holds universally. We studied species assemblages within the matrices of byssally interconnected mussels engineered by Mytilus edulis in the North Sea, by mixed Perumytilus purpuratus and Mytilus chilensis at the southern Chilean coast, by Musculista senhousia in the Yellow Sea and by Xenostrobus inconstans at the coast of southern Australia. In all cases, species assemblages inside mussel beds were significantly different from those outside with many species being restricted to one habitat type. However, species richness and diversity were not generally higher in mussel beds than in ambient sediments without mussels. In the North Sea (M. edulis) and at the Chilean coast (P. purpuratus, M. chilensis), mussel beds have markedly higher species numbers and diversities than surrounding sediments, but this was not the case for mussel beds in Australia (X. inconstans) and the Yellow Sea (M. senhousia) where numbers of associated species were only slightly higher and somewhat lower than in adjacent sediments, respectively. In conclusion, although soft bottom mytilid mussels generally enhance habitat heterogeneity and species diversity at the ecosystem level, mussel beds themselves are not universal centres of biodiversity, but the effects on associated species are site specific.     

Growth performance, carcass characteristics and meat quality of group-penned surgically castrated, immunocastrated (Improvac®) and entire male pigs and individually penned entire male pigs

The objective of the study was to compare growth performance, carcass characteristics, meat quality and fatty acid composition of the adipose tissue of group-penned barrows, immunocastrated boars and entire males. Furthermore, the effect of housing of entire males on the aforementioned parameters was evaluated. At 55.2 days of age, 52 Swiss Large White pigs were blocked by litter and assigned by BW to four experimental groups: barrows (C), immunocastrated boars (IC), entire males (EMG) reared in group pens and entire males (EMP) reared in individual pens. In experiment 1, the effects of the method of castration were investigated (experimental groups C, IC and EMG). In experiment 2, the effects of housing on entire male pigs were evaluated (experimental groups EMG and EMP). All pigs had ad libitum access to standard diets from weaning to 107 kg BW. The two vaccinations (Improvac®) were applied to the IC pigs at an average BW of 22.6 and 73.0 kg. In experiment 1, average daily gain (ADG) did not (P > 0.05) differ among the experimental groups. However, EMG consumed less feed and had a better feed-conversion ratio than C (P < 0.001 for each). For IC, intermediate values were observed, which differed (P < 0.001) from EMG and C. Lean meat percentage decreased (P < 0.05) from EMG to IC, and from IC to C. The androstenone and skatole levels were higher (P < 0.05) in the adipose tissue of EMG than IC and C. Shear force values were higher (P < 0.01) in the longissimus muscle of C and EMG, compared to IC. The concentration of saturated fatty acid in the adipose tissue increased (P < 0.001) from EMG to IC, and from IC to C pigs, and that of polyunsaturated fatty acid decreased (P < 0.001). In experiment 2, ADG did not (P > 0.05) differ between EMP and EMG. However, EMP pigs consumed more feed than EMG pigs and had a poorer feed efficiency (P < 0.01 for each). In conclusion, EMG pigs had a better feed efficiency than IC pigs and their carcasses were leaner, but the risk of boar tainted pork was elevated. Group-housing negatively affected average daily feed intake but not ADG of entire males. At the moment, immunocastration offers a good approach to avoid castration and minimize the risk of boar taint.     

Depressed Pollination of Lapageria rosea Ruiz et Pav. (Philesiaceae) in the Fragmented Temperate Rainforest of Southern South America

We studied the pollination and reproductive success in continuous and fragmented populations of Lapageria rosea, a self-compatible plant endemic to temperate forests of Chile. Pollinator abundance, visitation rates, flower abundance, nectar volume and concentration, pollen germination and fruit and seed production, were compared between continuous forest of 145 ha and four forest fragments of 6, 3, 3, and 1 ha respectively, surrounded by mature pine plantations of Pinus radiata. Flower abundance was lower in three out of four forest fragments relative to continuous forest. Nectar volume and sugar concentration did not differ between flowers in the two habitats. Pollinators of L. rosea, the hummingbird Sephanoides sephaniodes and bumblebee Bombus dahlbomii were less abundant and visited flowers of L. rosea at lower rates in fragments than in continuous forest. In addition, in vitro rates of pollen germination were lower for flowers in forest fragments. The number of seeds per fruit was also lower in forest fragments. We suggest that fragmentation affects the reproductive success of L. rosea, lowering the total numbers of seeds produced and possibly compromising long term persistence of fragmented populations.