Isolation of bacteria from fermented food and grass carp intestine and their efficiencies in improving nutrient value of soybean meal in solid state fermentation

Background: Soybean meal is an excellent and cost-effective protein source; however, its usage is limited in the piglet due to the presence of anti-nutritional factors and the antigens glycinin and β-conglycinin. The objective of the current study was to screen and select for bacteria that can be efficiently adopted to ferment soybean meal in order to solve this problem.Results: Bacteria were isolated from fermented soy foods and the grass carp intestine, and strains selected for high protease, cellulase and amylase activities. The isolated bacteria were characterized as Bacillus cereus, Bacillus subtilis and Bacilus amyloliquefacien, respectively. Fermentation with food-derived Isolate-2 and fish-derived F-9 increased crude protein content by 5.32% and 8.27%, respectively; improved the amino acid profile by increasing certain essential amino acids, broke down larger soy protein to 35 k Da and under, eliminated antigenicity against glycinin and β-conglycinin, and removed raffinose and stachyose in the soybean meal following a 24-h fermentation.Conclusions: Our results suggest these two B. amyloliquefaciens bacteria can efficiently solid state ferment soybean meal and ultimately produce a more utilizable food source for growing healthy piglets.     

Comment on "Calcareous nannoplankton response to surface-water acidification around Oceanic Anoxic Event 1a"

Erba et al. (Reports, 23 July 2010, p. 428) attributed calcareous nannofossil morphology and assemblage changes across Cretaceous Oceanic Anoxic Event 1a to the effects of surface ocean acidification. We argue that the quality of carbonate preservation in these sequences, the unsupported assumptions of the biotic response to acidity, and the absence of independent proxy estimates for ocean pH or atmospheric pCO(2) render this conclusion questionable.     

Developing Agreement Between Very Short Flow and Longer Flow Test Wheat Mills

Variations in soft wheat moisture content and kernel texture greatly affected the flour yield produced by a small (short flow) microtest mill (Quadrumat Jr.). An algorithm was developed that adjusted Quadrumat Jr. flour yield to 15% wheat moisture content, precluding the need to temper the wheat before milling. Another algorithm was developed to adjust Quadrumat flour yield relative to a constant softness equivalent (measurement of kernel texture) obtained during the micromilling procedure. Predicting the flour yield of the longer flow Allis‐Chalmers mill from Quadrumat Jr. unadjusted flour yield (R ² = 0.55) was compared with predicting Allis‐Chalmers flour yield from the Quadrumat Jr. adjusted flour yield (R ² = 0.90) across five diverse confirmation data sets. An algorithm to adjust flour yield for softness equivalent was individually developed for soft and hard wheats. Representative micromilling flour yield and softness equivalent data could be produced using as little as 10 g of untempered wheat and ≈3 min of operator time.     

Intragenic vectors for gene transfer without foreign DNA

The intragenic vector system involves identifying functional equivalents of vector components from the genome of a specific crop species (or related species to which it can be hybridised) and using these DNA sequences to assemble vectors for transformation of that plant species. This system offers an attractive alternative to current genetic engineering strategies where vectors are based on DNA sequences that usually originate from bacteria. The construction of intragenic vectors enables the well-defined genetic improvement of plants with all transferred DNA originating from within the gene pool already available to plant breeders. In this manner genes can be introgressed into elite cultivars in a single step without linkage drag and without the incorporation of foreign DNA. The resulting plants are non-transgenic, although they are derived using the tools of molecular biology and plant transformation. The use of intragenic vectors for the transfer of genes from within the gene pools of crops may help to alleviate some of the major public concerns over the deployment of GM crops in agriculture, notably the ethical issue associated with the transfer of DNA across wide taxonomic boundaries. This paper reviews the progress toward the development and use of intragenic vectors and the implications of their use for the genetic improvement of crops. Dedicated to the late Hans Sandbrink for his enthusiasm in helping to develop intragenic vectors     

Population cycles in small rodents

We conclude that population fluctuations in Microtus in southern Indiana are produced by a syndrome of changes in birth and death rates similar to that found in other species of voles and lemmings. The mechanisms which cause the changes in birth and death rates are demolished by fencing the population so that no dispersal can occur. Dispersal thus seems critical for population regulation in Microtus. Because most dispersal occurs during the increase phase of the population cycle and there is little dispersal during the decline phase, dispersal is not directly related to population density. Hence the quality of dispersing animals must be important, and we have found one case of increased dispersal tendency by one genotype. The failure of population regulation of Microtus in enclosed areas requires an explanation by any hypothesis attempting to explain population cycles in small rodents. It might be suggested that the fence changed the predation pressure on the enclosed populations. However, the fence was only 2 feet (0.6 meter) high and did not stop the entrance of foxes, weasels, shrews, or avian predators. A striking feature was that the habitat in the enclosures quickly recovered from complete devastation by the start of the spring growing season. Obviously the habitat and food quality were sufficient to support Microtus populations of abnormally high densities, and recovery of the habitat was sufficiently quick that the introduction of new animals to these enclosed areas resulted in another population explosion. Finally, hypotheses of population regulation by social stress must account for the finding that Microtus can exist at densities several times greater than normal without "stress" taking an obvious toll. We hypothesize that the prevention of dispersal changes the quality of the populations in the enclosures in comparison to those outside the fence. Voles forced to remain in an overcrowded fenced population do not suffer high mortality rates and continue to reproduce at abnormally high densities until starvation overtakes them. The initial behavioral interactions associated with crowding do not seem sufficient to cause voles to die in situ. What happens to animals during the population decline? Our studies have not answered this question. The animals did not appear to disperse, but it is possible that the method we used to measure dispersal (movement into a vacant habitat) missed a large segment of dispersing voles which did not remain in the vacant area but kept on moving. Perhaps the dispersal during the increase phase of the population cycle is a colonization type of dispersal, and the animals taking part in it are likely to stay in a new habitat, while during the population decline dispersal is a pathological response to high density, and the animals are not attracted to settling even in a vacant habitat. The alternative to this suggestion is that animals are dying in situ during the decline because of physiological or genetically determined behavioral stress. Thus the fencing of a population prevents the change in rates of survival and reproduction, from high rates in the increase phase to low rates in the decline phase, and the fenced populations resemble "mouse plagues." A possible explanation is that the differential dispersal of animals during the phase of increase causes the quality of the voles remaining at peak densities in wild populations to be different from the quality of voles at much higher densities in enclosures. Increased sensitivity to density in Microtus could cause the decline of wild populations at densities lower than those reached by fenced populations in which selection through dispersal has been prevented. Fencing might also alter the social interactions among Microtus in other ways that are not understood. The analysis of colonizing species by MacArthur and Wilson (27) can be applied to our studies of dispersal in populations of Microtus. Groups of organisms with good dispersal and colonizing ability are called r strategists because they have high reproductive potential and are able to exploit a new environment rapidly. Dispersing voles seem to be r strategists. Young females in breeding condition were over-represented in dispersing female Microtus (17). The Tf(C)/Tf(E) females, which were more common among dispersers during the phase of population increase (Fig. 6), also have a slight reproductive advantage over the other Tf genotypes (19). Thus in Microtus populations the animals with the highest reproductive potential, the r strategists, are dispersing. The segment of the population which remains behind after the selection-via-dispersal are those individuals which are less influenced by increasing population densities. These are the individuals which maximize use of the habitat, the K strategists in MacArthur and Wilson's terminology, or voles selected for spacing behavior. Thus we can describe population cycles in Microtus in the same theoretical framework as colonizing species on islands. Our work on Microtus is consistent with the hypothesis of genetic and behavioral effects proposed by Chitty (6) (Fig. 7) in that it shows both behavioral differences in males during the phases of population fluctuation and periods of strong genetic selection. The greatest gaps in our knowledge are in the area of genetic-behavioral interactions which are most difficult to measure. We have no information on the heritability of aggressive behavior in voles. The pathways by which behavioral events are translated into physiological changes which affect reproduction and growth have been carefully analyzed by Christian and his associates (28) for rodents in laboratory situations, but the application of these findings to the complex field events described above remains to be done. Several experiments are suggested by our work. First, other populations of other rodent species should increase to abnormal densities if enclosed in a large fenced area (29). We need to find situations in which this prediction is not fulfilled. Island populations may be an important source of material for such an experiment (30). Second, if one-way exit doors were provided from a fenced area, normal population regulation through dispersal should occur. This experiment would provide another method by which dispersers could be identified. Third, if dispersal were prevented after a population reached peak densities, a normal decline phase should occur. This prediction is based on the assumption that dispersal during the increase phase is sufficient to ensure the decline phase 1 or 2 years later. All these experiments are concerned with the dispersal factor, and our work on Microtus can be summarized by the admonition: study dispersal.     

Effect of growth hormone overexpression on gastric evacuation rate in coho salmon

Growth hormone (GH) transgenic (T) coho salmon consistently show remarkably enhanced growth associated with increased appetite and food consumption compared to non-transgenic wild-type (NT) coho salmon. To improve understanding of the mechanism by which GH overexpression mediates food intake and digestion in T fish, feed intake and gastric evacuation rate (over 7 days) were measured in size-matched T and NT coho salmon. T fish displayed greatly enhanced feed intake levels (~ 2.5-fold), and more than 3-fold increase in gastric evacuation rates relative to NT coho salmon. Despite the differences in feed intake, no differences were noted in the time taken from first ingestion of food to stomach evacuation between genotypes. These results indicate that enhanced feed intake is coupled with an overall increased processing rate to enhance energy intake by T fish. To further investigate the molecular basis of these responses, we examined the messenger RNA (mRNA) levels of several genes in appetite- and gastric-regulation pathways (Agrp1, Bbs, Cart, Cck, Glp, Ghrelin, Grp, Leptin, Mc4r, Npy, and Pomc) by qPCR analyses in the brain (hypothalamus, preoptic area) and pituitary, and in peripheral tissues associated with digestion (liver, stomach, intestine, and adipose tissue). Significant increases in mRNA levels were found for Agrp1 in the preoptic area (POA) of the brain, and Grp and Pomc in pituitary for T coho salmon relative to NT. Mch and Npy showed significantly lower mRNA levels than NT fish in all brain tissues examined across all time-points after feeding. Mc4r and Cart for T showed significantly lower mRNA levels than NT in the POA and hypothalamus, respectively. In the case of peripheral tissues, T fish had lower mRNA levels of Glp and Leptin than NT fish in the intestine and adipose tissue, respectively. Grp, Cck, Bbs, Glp, and Leptin in stomach, adipose tissue, and/or intestine showed significant differences across the time-points after feeding, but Ghrelin showed no significant difference between T and NT fish in all tested tissues.     

Recruitment dynamics in complex life cycles

Organisms living in the marine rocky intertidal zone compete for space. This, together with predation, physical disruption, and differing species tolerances to physiological stress, explains the structure of the ecological communities at some sites. At other sites the supply of larvae is limiting, and events in the offshore waters, such as wind-driven upwelling, explain the composition of intertidal communities. Whether the community ecology at a site is governed by adult-adult interactions within the site, or by limitations to the supply of larvae reaching the site, is determined by the regional pattern of circulation in the coastal waters. Models combining larval circulation with adult interactions can potentially forecast population fluctuations. These findings illustrate how processes in different ecological habitats are coupled.     

Diverse and contrasting effects of habitat fragmentation

Different components of an ecosystem can respond in very different ways to habitat fragmentation. An archipelago of patches, representing different levels of fragmentation, was arrayed within a successional field and studied over a period of 6 years. Ecosystem processes (soil mineralization and plant succession) did not vary with the degree of subdivision, nor did most measures of plant and animal community diversity. However, fragmentation affected vertebrate population dynamics and distributional patterns as well as the population persistence of clonal plant species. The results highlight the dangers of relying on broad community measures in lieu of detailed population analyses in studies of fragmented habitats.     

Escherichia coli isolated from bovine mastitis invade mammary cells by a modified endocytic pathway

Escherichia coli is a major pathogen in the aetiology of bovine mastitis. Although classically considered to be an environmental pathogen causing mainly transient infection, the incidence of persistent E. coli mastitis infections may be increasing, suggesting an adaptation of this pathogen to the bovine udder environment. Mastitis E. coli strains have been demonstrated to enter bovine mammary cells in vitro but little is known about the invasion mechanism or the intracellular fate of the bacteria. In order to further understand the pathogenesis of persistent E. coli bovine mastitis we investigated the intracellular trafficking of mastitis E. coli isolates in primary bovine mammary cells using confocal microscopy and fluorescent markers of endocytic compartments. Consistent with other studies, mastitis E. coli were found to invade primary bovine mammary cells in vitro. This process did not involve in the rearrangement of the actin cytoskeleton. Intracellular bacteria were observed within membrane-bound compartments that labelled with the early endosomal marker phosphatidylinositol 3-phosphate (PtdIns(3)P) and also within late endosome-like compartments labelled with the small GTPase Rab7, indicating an endocytic mechanism of bacterial internalization. Bacteria were not observed within acidified lysosomal compartments or autophagic vacuoles, suggesting that the internalized bacteria are not targeted for lysosomal degradation via either the classical endocytic pathway or the autophagic response. Our findings are consistent with an endosomal survival niche for the internalized bacteria, allowing them to evade host immune responses and establish an infection reservoir that could later re-emerge as a recurrent clinical mastitis episode.