Evaluation of Phytate-Degrading Lactobacillus Culture Administration to Broiler Chickens

Probiotics have been demonstrated to promote growth, stimulate immune responses, and improve food safety of poultry. While widely used, their effectiveness is mixed, and the mechanisms through which they contribute to poultry production are not well understood. Microbial phytases are increasingly supplemented in feed to improve digestibility and reduce antinutritive effects of phytate. The microbial origin of these exogenous enzymes suggests a potentially important mechanism of probiotic functionality. We investigated phytate degradation as a novel probiotic mechanism using recombinant Lactobacillus cultures expressing Bacillus subtilis phytase. B. subtilis phyA was codon optimized for expression in Lactobacillus and cloned into the expression vector pTRK882. The resulting plasmid, pTD003, was transformed into Lactobacillus acidophilus, Lactobacillus gallinarum, and Lactobacillus gasseri. SDS-PAGE revealed a protein in the culture supernatants of Lactobacillus pTD003 transformants with a molecular weight similar to that of the B. subtilis phytase. Expression of B. subtilis phytase increased phytate degradation of L. acidophilus, L. gasseri, and L. gallinarum approximately 4-, 10-, and 18-fold over the background activity of empty-vector transformants, respectively. Phytase-expressing L. gallinarum and L. gasseri were administered to broiler chicks fed a phosphorus-deficient diet. Phytase-expressing L. gasseri improved weight gain of broiler chickens to a level comparable to that for chickens fed a control diet adequate in phosphorus, demonstrating proof of principle that administration of phytate-degrading probiotic cultures can improve performance of livestock animals. This will inform future studies investigating whether probiotic cultures are able to provide both the performance benefits of feed enzymes and the animal health and food safety benefits traditionally associated with probiotics.     

Staphylococcus epidermidis in the human skin microbiome mediates fermentation to inhibit the growth of Propionibacterium acnes: implications of probiotics in acne vulgaris

Increasing evidence demonstrates that commensal microorganisms in the human skin microbiome help fight pathogens and maintain homeostasis of the microbiome. However, it is unclear how these microorganisms maintain biological balance when one of them overgrows. The overgrowth of Propionibacterium acnes (P. acnes), a commensal skin bacterium, has been associated with the progression of acne vulgaris. Our results demonstrate that skin microorganisms can mediate fermentation of glycerol, which is naturally produced in skin, to enhance their inhibitory effects on P. acnes growth. The skin microorganisms, most of which have been identified as Staphylococcus epidermidis (S. epidermidis), in the microbiome of human fingerprints can ferment glycerol and create inhibition zones to repel a colony of overgrown P. acnes. Succinic acid, one of four short-chain fatty acids (SCFAs) detected in fermented media by nuclear magnetic resonance (NMR) analysis, effectively inhibits the growth of P. acnes in vitro and in vivo. Both intralesional injection and topical application of succinic acid to P. acnes-induced lesions markedly suppress the P. acnes-induced inflammation in mice. We demonstrate for the first time that bacterial members in the skin microbiome can undergo fermentation to rein in the overgrowth of P. acnes. The concept of bacterial interference between P. acnes and S. epidermidis via fermentation can be applied to develop probiotics against acne vulgaris and other skin diseases. In addition, it will open up an entirely new area of study for the biological function of the skin microbiome in promoting human health.     

Linking Aβ42-Induced Hyperexcitability to Neurodegeneration,Learning and Motor Deficits,and a Shorter Lifespan in an Alzheimer’s Model

Alzheimer’s disease (AD) is the most prevalent form of dementia in the elderly. β-amyloid (Aβ) accumulation in the brain is thought to be a primary event leading to eventual cognitive and motor dysfunction in AD. Aβ has been shown to promote neuronal hyperactivity, which is consistent with enhanced seizure activity in mouse models and AD patients. Little, however, is known about whether, and how, increased excitability contributes to downstream pathologies of AD. Here, we show that overexpression of human Aβ42 in a Drosophila model indeed induces increased neuronal activity. We found that the underlying mechanism involves the selective degradation of the A-type K+ channel, Kv4. An age-dependent loss of Kv4 leads to an increased probability of AP firing. Interestingly, we find that loss of Kv4 alone results in learning and locomotion defects, as well as a shortened lifespan. To test whether the Aβ42-induced increase in neuronal excitability contributes to, or exacerbates, downstream pathologies, we transgenically over-expressed Kv4 to near wild-type levels in Aβ42-expressing animals. We show that restoration of Kv4 attenuated age-dependent learning and locomotor deficits, slowed the onset of neurodegeneration, and partially rescued premature death seen in Aβ42-expressing animals. We conclude that Aβ42-induced hyperactivity plays a critical role in the age-dependent cognitive and motor decline of this Aβ42-Drosophila model, and possibly in AD.     

Pollen-mediated gene flow in isolated and continuous stands of bur oak, Quercus macrocarpa (Fagaceae)

? Premise of the study: Pollination patterns determine the reproductive neighborhood size of plants, the connectivity of populations, and the impacts of habitat fragmentation. We characterized pollination in three populations of Quercus macrocarpa occurring in a highly altered landscape in northeastern Illinois to determine whether isolated remnant stands were reproductively isolated. ? Methods: We used microsatellites to genotype all adults and 787 acorns from two isolated savanna remnants and a stand in an old-growth forest. One isolated remnant occurred in a highly urbanized/industrialized landscape, and one occurred in an agricultural landscape. Parentage assignment was used to assess pollen-mediated gene flow. ? Key results: Pollen donors from outside the study sites accounted for between 46% and 53% of paternities and did not differ significantly among sites, indicating that similar high levels of gene flow occurred at all three sites. Within stands, the mean pollination distance ranged from 42 to 70 meters, and when accounting for outside pollinations, mean pollination distances were well over 100 meters. Genetic diversity of incoming pollen was extremely high in all three stands. The number of effective pollen donors, N(ep), calculated from paternity assignment was higher than that estimated by an indirect correlated paternity approach. ? Conclusions: Our findings indicate that extremely isolated stands of oaks are unlikely to be genetically and reproductively isolated, and remnant stands may contribute to maintaining genetic connectivity in highly modified landscapes.     

Retention of a transgenerational marker (137Barium) in tissues of adult female anemonefish and assessment of physiological stress

Recently it was shown that female fish injected with enriched stable isotopes maternally transmit a chemical signature to larval otoliths. Validation of this larval marking technique requires laboratory experiments to determine appropriate injection concentrations and to assess any negative effects on larval and adult condition. This study investigated the temporal profile of ¹³⁷barium assimilation and retention in tissues of adult female anemonefish Amphiprion melanopus (Pomacentridae) following intraperitoneal injection with either 2 or 4 μg ¹³⁷Ba g^?1 body mass. Mean barium isotope ratios (¹³⁸Ba:¹³⁷Ba) in the two groups of treated fish were not significantly different from each other, but were significantly different from those in the control group up to 56 days post-injection. This pattern of ¹³⁷Ba retention was consistent across gonad, muscle, liver and bone tissues. Mean plasma cortisol concentration (an indicator of non-specific physiological stress) was not significantly different among groups and was considered to be representative of unstressed fish. Together, these results indicate that (1) A. melanopus suffer minimal physiological stress and cope well after treatment with ¹³⁷Ba, (2) ¹³⁷Ba is retained in female A. melanopus for a prolonged period (at least 56 days), such that multiple clutches of offspring are likely to be marked with an isotopic signature, and (3) a lower dosage of 2 μg ¹³⁷Ba g^?1 appears sufficient for transgenerational marking. It is concluded that ¹³⁷Ba is suitable for use as a transgenerational marker and is a powerful tool to resolve long-standing enigmas such as larval dispersal distances and the fishery benefits of marine reserves.     

The effectiveness of rotating brush devices for management of vessel hull fouling

The present study tested two diver-operated rotating brush systems, coupled with suction and collection capabilities, to determine their efficacy in the management of vessel biofouling. Both rotating brush systems proved effective (>80%) in removing low-to-moderate levels of fouling from flat and curved experimental surfaces (Perspex plates). However, performance was generally poorer at removing more advanced levels of fouling. In particular, mature calcareous organisms were relatively resistant to the rotating brushes, with a high proportion (up to 50%) remaining on plates following treatment. On average, >95% of defouled material was collected and retained by both systems. The amount of lost material generally increased when treating curved plates with increasing biomass, whereas the material lost from flat plates was typically less and remained relatively constant throughout the trials. The majority (>80%) of fouling not captured by the systems was crushed by the brushes (ie non-viable). However, a diverse range of viable organisms (eg barnacles and hydroids) was lost to the environment during the defouling trials. When defouling a vessel, unintentional detachment of fouling organisms is likely to be high through physical disturbance by divers operating the devices and by associated equipment (eg hoses). Furthermore, residual biosecurity risks are also likely to remain due to diver error, persistent fouling remaining on treated surfaces and the inaccessibility of niche areas to the brush systems. To address these limitations, further research into alternative treatment methods is required.     

Metal ions,pH, and cholesterol regulate the interactions of Alzheimer's disease amyloid-beta peptide with membrane lipid

The interaction of A beta peptides with the lipid matrix of neuronal cell membranes plays an important role in the pathogenesis of Alzheimer's disease. By using EPR and CD spectroscopy, we found that in the presence of Cu(2+) or Zn(2+), pH, cholesterol, and the length of the peptide chain influenced the interaction of these peptides with lipid bilayers. In the presence of Zn(2+), A beta 40 and A beta 42 both inserted into the bilayer over the pH range 5.5-7.5, as did A beta 42 in the presence of Cu(2+). However, A beta 40 only penetrated the lipid bilayer in the presence of Cu(2+) at pH 5.5-6.5; at higher pH there was a change in the Cu(2+) coordination sphere that inhibited membrane insertion. In the absence of the metals, insertion of both peptides only occurred at pH < 5.5. Raising cholesterol to 0.2 mol fraction of the total lipid inhibited insertion of both peptides under all conditions investigated. Membrane insertion was accompanied by the formation of alpha-helical structures. The nature of these structures was the same irrespective of the conditions used, indicating a single low energy structure for A beta in membranes. Peptides that did not insert into the membrane formed beta-sheet structures on the surface of the lipid.     

Phylogeny and Classification of the Trapdoor Spider Genus Myrmekiaphila: An Integrative Approach to Evaluating Taxonomic Hypotheses

Background

Revised by Bond and Platnick in 2007, the trapdoor spider genus Myrmekiaphila comprises 11 species. Species delimitation and placement within one of three species groups was based on modifications of the male copulatory device. Because a phylogeny of the group was not available these species groups might not represent monophyletic lineages; species definitions likewise were untested hypotheses. The purpose of this study is to reconstruct the phylogeny of Myrmekiaphila species using molecular data to formally test the delimitation of species and species-groups. We seek to refine a set of established systematic hypotheses by integrating across molecular and morphological data sets.

Methods and Findings

Phylogenetic analyses comprising Bayesian searches were conducted for a mtDNA matrix composed of contiguous 12S rRNA, tRNA-val, and 16S rRNA genes and a nuclear DNA matrix comprising the glutamyl and prolyl tRNA synthetase gene each consisting of 1348 and 481 bp, respectively. Separate analyses of the mitochondrial and nuclear genome data and a concatenated data set yield M. torreya and M. millerae paraphyletic with respect to M. coreyi and M. howelli and polyphyletic fluviatilis and foliata species groups.

Conclusions

Despite the perception that molecular data present a solution to a crisis in taxonomy, studies like this demonstrate the efficacy of an approach that considers data from multiple sources. A DNA barcoding approach during the species discovery process would fail to recognize at least two species (M. coreyi and M. howelli) whereas a combined approach more accurately assesses species diversity and illuminates speciation pattern and process. Concomitantly these data also demonstrate that morphological characters likewise fail in their ability to recover monophyletic species groups and result in an unnatural classification. Optimizations of these characters demonstrate a pattern of “Dollo evolution” wherein a complex character evolves only once but is lost multiple times throughout the group''s history.