In vitro assessment of three fibrolytic enzyme preparations as potential feed additives in equine diets

A series of in vitro experiments were conducted to assess three fibrolytic enzyme preparations as potential feed additives in equine diets. The three fibrolytic enzyme preparations were a concentrated cellulase (E1), an acid cellulase (E2) and a concentrated xylanase (E3). The enzymes were evaluated on their ability to modify the cell wall fraction of high-temperature dried lucerne (HTL) under various experimental conditions including differences in temperature, pH, incubation period, substrate levels and particle size to enable selection of the enzyme preparation most effective in the hydrolysis of lucerne. Results showed enzyme activities (as measured by reducing sugar assays) to be greatest at 50 °C, pH 5 and over an incubation period of greater than 20 h. E1 exhibited the greatest effect on total monosaccharide release from the HTL compared to E2 and E3. Moreover, dry matter (DM) and total non-starch polysaccharide (TNSP) losses were also greater in HTL treated with E1 compared to E2 and E3. Therefore, since the cell wall fraction of HTL contained substantial amounts of cellulose, the enzyme with the highest cellulase activity (Enzyme 1) was most effective in hydrolysing the cell walls of HTL. Consequently, it would appear that the application of exogenous fibrolytic enzyme preparations to forages requires the chemical characterisation of the target forage to enable selection of enzymes that are (a) most suitable to degrade the cell wall components of the candidate forage and (b) effective under field conditions.     

The Amazon River as a dispersal barrier to passerine birds: effects of river width, habitat and taxonomy

Aim To evaluate the relative effectiveness of the lower and upper sections, respectively, of the Amazon River as a barrier to bird distribution, and to evaluate ecological and taxonomic factors affecting the efficacy of the river barrier. Location Amazon River of South America between its confluence with the Napo River in the west and its delta in the east. Methods Using published distribution maps for 448 species of passerine birds occurring along the Amazon River, we evaluated whether each was distributed along one bank only (river presumed to be a barrier) or both banks (no barrier) to test the predictions that the river was more effective as a dispersal barrier: (1) along the lower, wider portion of the river than the upper, narrower portion; (2) for species inhabiting forests than open country; (3) for species inhabiting forest understorey than forest canopy; (4) for species restricted to terra firme (never inundated upland forest) than those not restricted to terra firme and (5) for certain taxonomic groups. Results Our analyses demonstrated that the Amazon River was most effective as a dispersal barrier along its lower portion and for species restricted to forests and terra firme. However, the river was not significantly more of a barrier for species inhabiting forest understorey than forest canopy. The river was most significant as a barrier to dispersal for the antbirds (Thamnophilidae) and was less significant as a barrier to species belonging to several large families including woodcreepers (Dendrocolaptidae), ovenbirds (Furnariidae), flycatchers (Tyrannidae), cotingids (Cotingidae), tanagers (Thraupidae), seed‐eating finches (Emberizidae) and blackbirds (Icteridae). Main conclusions The robust widths of Amazonian rivers are widely considered to represent impediments to dispersal and gene flow for many taxa of birds and other animals, and may have represented agents of vicariance in the diversification of species. Our study reaffirms the effectiveness of the lower Amazon River as a current barrier to bird dispersal for forest birds and provides new insights into the effects of habitat and taxonomy on the efficacy of the river barrier. Although supportive of several predictions of the river hypothesis of biological diversification, our study is limited in addressing the historical impact of river barriers as agents of vicariance in the process of diversification.     

Attenuated virulence of min operon mutants of Neisseria gonorrhoeae and their interactions with human urethral epithelial cells

Neisseria gonorrhoeae, a sexually-transmitted gram-negative bacterium, causes gonorrhoea in humans. The min genes of N. gonorrhoeae are involved in cell division site selection with oxyR co-transcribed with these genes. The mutation in min genes and oxy R cause aberrant cell morphology and aggregation patterns, respectively. Our objective was to assess the contribution of neisserial min operon cell division genes i.e. minC, minD and oxyR in virulence. Compared to the N. gonorrhoeae parental strain (Ng CH811Str(R)), its isogenic mutants with insertionally inactivated minC (Ng CSRC1), minD (Ng CJSD1) or oxyR (Ng KB1) showed reduced adherence to and invasion of urethral epithelial cells. This may be explained by defective microcolony formation in the mutant strains, possibly owing to abnormal morphology and aggregation. The expression levels of surface virulence factors like Opa, pilin and lipooligosaccharide in the mutants were unchanged relative to Ng CH811Str(R). Furthermore, in urethral epithelial cells, the min and oxyR mutants induced the release of proinflammatory cytokines like IL6 and IL8 to levels similar to that induced by the parental strain. Taken together, our studies indicate that inactivation of minC, minD or oxyR in N. gonorrhoeae attenuates its ability to bind to and invade urethral epithelial cells without altering its potential to induce IL6 and IL8 release.     

Multiple Origins and Replication Proteins Influence Biological Properties of β-Lactamase-Producing Plasmids from Neisseria gonorrhoeae

The beta-lactamase-producing Asia-type plasmid pJD4 of Neisseria gonorrhoeae is a 7.4-kb, broad-host-range plasmid. It is part of a family of plasmids which are structurally related yet vary in size, found in both N. gonorrhoeae and Haemophilus ducreyi. Branch-point analysis by electron microscopy indicates that pJD4 carries three clustered but distinguishable origins of replication, which we named ori1, ori2, and ori3. Although pJD4 belongs to incompatibility (Inc) group W, it also carries a silent IncFII determinant which is expressed when ori2 and ori3 are absent. The Africa-type plasmid pJD5, a naturally occurring deletion derivative of pJD4, carries only ori1, belongs to the IncFII group, and, in contrast to pJD4, requires DNA polymerase I (Pol I) for replication. Plasmids constructed from pJD4 which lack ori1 but carry ori2 and ori3 do not require Pol I and are incompatible with IncW plasmids, suggesting that the ori2 or ori3 region contains the IncW determinant. We have cloned a replication initiation protein (RepB) that is necessary for ori2 and ori3 to function. This Rep protein is distinct from RepA, which is necessary for ori1. Thus, pJD4 is unique because it is the smallest plasmid characterized containing three origins of replication and two unique Rep proteins.     

Communities of constructional lips and cup reef rims in Bermuda

Constructional lips and cup reef rims are little studied reef features typical of very turbulent conditions. The relatively low diversity community of these structures consists of three components, and algal mat dominated by Herposiphonia secunda, an encrusting calcareous biota characterised by crustose coralline algae and the vermetid gastropod Dendropoma corrodens, which together are the main hermatypic components, and an endolithic component with very abundant boring sponges, polychaetes, crustaceans and sipunculids. In northerly areas of Bermuda, the hydrozoan Millepora alcicornis is a prominent member of the community while to the south the urchin Echinometra lucunter is abundant. On the south side of Bermuda the community traps sediment which is deposited in the void spaces. The environment shows reduced grazing and this may account for the presence of several relict and rare species.     

Effect of bacterial vaginosis, Lactobacillus and Premarin estrogen replacement therapy on vaginal gene expression changes

The aim of the study was to investigate gene expression profiles of post-menopausal women receiving Premarin estrogen replacement therapy (ERT), compared to controls, and to examine any correlations between the bacterial vaginosis (BV) status of the subjects. Based upon an expected finding of a 50-60% difference between gene expression of host antimicrobials with alpha=0.05 (2-sided), beta=0.20 the calculation of 7 subjects per group, led to a sample size of 10 subjects receiving Premarin estrogen replacement therapy and 10 healthy, age-matched controls. Vaginal samples were collected at a single timepoint and processed for RNA recovery and Affymetrix array analysis, as well as Nugent scoring and denaturing gradient gel electrophoresis to identify bacteria. Lactobacillus iners was the most commonly detected species in the normal flora and this was confirmed with L. iners-specific PCR method. Vaginal swabs from 6 Premarin and 8 control vaginal samples provided a non-invasive means to analyze human gene expression. There was no significant up-regulation of cancer-associated gene expression in subject receiving Premarin ERT, but some evidence that the potentially protective innate immunity was reduced in patients with BV. Of those with a normal flora, there was a 2-fold down-regulation of carcinoma associated forkhead box A1 gene expression. BV was associated with 7-fold down-regulation of host antimicrobial colony stimulating factor, -9.83-fold for IL-1alpha, -8.33 for IL-1beta and -3.63 for IL-6. This is the first study to use gene arrays to correlate changes in host expression response to estrogen replacement therapy and BV.     

<Emphasis Type="Italic">Plectosphaerella cucumerina</Emphasis> as a bioherbicide for <Emphasis Type="Italic">Cirsium arvense</Emphasis>: proof of concept

Plectosphaerella cucumerina (Lindf.) W. Gams was evaluated as a bioherbicide for Cirsium arvense L. (Scop.) using a Canadian and a New Zealand isolate. Both isolates defoliated C. arvense when applied at 10¹³ conidia ha^?1 in water volumes ranging from 250 to 6400 l ha^?1 with a rapid decline in effect with declining conidial dose. Repeat application and the addition of the adjuvant Pulse^® penetrant to the conidial suspension increased the disease severity in C. arvense. Maximum disease occurred at 20 °C with a 48 h post-application dew period. The experiments demonstrate that P. cucumerina can defoliate C. arvense under the environmental conditions of temperate pastures where the weed is problematic. The results also show that modifications to formulation and strategic application may reduce the 48 h dew period requirement and risk to non-target species respectively, supporting the conclusion that the fungus has potential as a bioherbicide for C. arvense.     

Amplification of DNA Encoding Entire Type I Polyketide Synthase Domains and Linkers from Streptomyces Species

Polyketides are a group of bioactive compounds from bacteria, plants, and fungi. To increase the availability of analogs for testing, the active sites of polyketide synthases are often substituted with homologous domains having altered substrate specificities. This study reports the design of polymerase chain reaction primers that enables isolation of entire active site domains from type I polyketide synthases with native interdomain linkers. This bypasses the need for further genetic screening to obtain functional units for use in genetic engineering. This is especially important in bioprospecting projects exploring new environments for bioresources.     

Abundance of lamprey larvae and physical habitat

Synopsis Southern brook lamprey larvae,Ichthyomyzon gagei, were collected from four creeks in Alabama and Mississippi. Larvae abundance (number m^–2) and length (mm) were measured over a range of sites to quantify some of the physical parameters involved in larval habitat selection. Particle size distribution, hydraulic conductivity, porosity and organic content were the factors analyzed. Abundance of larvae was greatest when particles smaller than 0.15 mm and 1.–2.0 mm in diameter represented at least 40% and 8% dry weight of the substrate, respectively. Abundance of larvae was lowest when the small particles represented less than 10% dry weight of the substrate. A strong inverse relationship was observed between density and length of larvae. All other parameters did not significantly affect lamprey habitat selection. Results of this study may be applicable in directing attempts at aquatic habitat restoration for lamprey in endangered streams.