Comparison of Sewage and Animal Fecal Microbiomes by Using Oligotyping Reveals Potential Human Fecal Indicators in Multiple Taxonomic Groups

Most DNA-based microbial source tracking (MST) approaches target host-associated organisms within the order Bacteroidales, but the gut microbiota of humans and other animals contain organisms from an array of other taxonomic groups that might provide indicators of fecal pollution sources. To discern between human and nonhuman fecal sources, we compared the V6 regions of the 16S rRNA genes detected in fecal samples from six animal hosts to those found in sewage (as a proxy for humans). We focused on 10 abundant genera and used oligotyping, which can detect subtle differences between rRNA gene sequences from ecologically distinct organisms. Our analysis showed clear patterns of differential oligotype distributions between sewage and animal samples. Over 100 oligotypes of human origin occurred preferentially in sewage samples, and 99 human oligotypes were sewage specific. Sequences represented by the sewage-specific oligotypes can be used individually for development of PCR-based assays or together with the oligotypes preferentially associated with sewage to implement a signature-based approach. Analysis of sewage from Spain and Brazil showed that the sewage-specific oligotypes identified in U.S. sewage have the potential to be used as global alternative indicators of human fecal pollution. Environmental samples with evidence of prior human fecal contamination had consistent ratios of sewage signature oligotypes that corresponded to the trends observed for sewage. Our methodology represents a promising approach to identifying new bacterial taxa for MST applications and further highlights the potential of the family Lachnospiraceae to provide human-specific markers. In addition to source tracking applications, the patterns of the fine-scale population structure within fecal taxa suggest a fundamental relationship between bacteria and their hosts.     

The effects of methyl palmitate,a putative regulator from perivascular fat,on the contractility of pregnant human myometrium

Aims

Methyl palmitate is thought to cause relaxation in vascular smooth muscle by opening voltage-activated potassium channels. We have tested the hypothesis that methyl palmitate, a putative regulator from perivascular fat, is an inhibitor of the contractility of human pregnant myometrium and that its effects might partially explain the higher incidence of dysfunctional labor in obese women compared to those with normal body mass indices.

Main methods

Strips of myometrium obtained with informed consent from women undergoing elective cesarean section at term were mounted in organ baths. Strips stimulated with oxytocin (1 nM) or KCl (30 mM) were exposed to cumulatively increasing concentrations of methyl palmitate up to 10 μM. Similar strips were exposed to cumulative addition of the potassium channel blockers 4-aminopyridine and tetraethylammonium. The contractility of the strips was monitored and analyzed using conventional methods.

Key findings

Methyl palmitate failed to inhibit oxytocin- or KCl-induced contractions over the concentration range tested. In fact, it exerted a slight excitatory effect in the presence of KCl, though not in the presence of oxytocin. The contractility of naïve strips was unaltered by exposure to 1 μM methyl palmitate. Both 4-aminopyridine and tetraethylammonium produced concentration-dependent contractions of human pregnant myometrium providing pharmacological evidence for the presence of voltage-activated potassium channels in this preparation.

Significance

Our findings do not support the hypothesis that methyl palmitate is an inhibitor of human pregnant myometrial contractility. Alternate hypotheses must be pursued to explain the higher incidence of dysfunctional labor in obese women.     

The effect of patient positioning on the precision of model-based radiostereometric analysis

A repeatable method for in vivo and in vitro measurement of polyethylene wear in total knee replacement (TKA) is needed. This research examines the model-based radiostereometric analysis’ (MBRSA) in vitro precision under different patient-radiograph orientations and flexion angles of the knee using a TKA phantom. Anterior–posterior and medial–lateral imaging orientations showed the highest precision; better than 0.036 mm (3-dimensional translation) and 0.089° (3-dimensional rotation). Flexion of the knee did not affect MBRSA precision. Medial–lateral imaging is advantageous as it allows for flexion of the knee joint during an RSA examination, thus providing greater information for wear measurement.     

Contribution of allochthonous resources to breeding in a high-arctic avian predator

Allochthonous input of resources (i.e., originating from a place other than where they are found) can have a significant impact on food availability for consumers. We assessed the impact of an allochthonous source of food (the sewage outfall stream of a military base) on an avian predator breeding in a low-productivity, high-arctic site (Alert, 83°N, 62°W, Ellesmere Island), the long-tailed skua (Stercorarius longicaudus). We collected blood samples throughout the breeding season during two contrasting years of lemming abundance to characterize diet composition of skuas and evaluated the contribution of the anthropogenic and lemming food sources using stable isotopes (carbon ¹³C and nitrogen ¹⁵N). The isotopic signature of skuas changed seasonally because they switch from a marine to a terrestrial diet when they come ashore to breed but also differed between the 2 years of the study. Anthropogenic food source accounted for 33 % of the summer diet but this proportion varied between years, from 41 % (5–95 ‰: 13–59 %) in a year of low lemming abundance to 16 % (5–95 ‰: 10–21 %) in a high year. Skua nest density recorded in years of low lemming abundance at Alert (0.15 nests/km²) was higher than at any other comparable arctic sites (0–0.02 nests/km²). Overall, the use of an anthropogenic food sources apparently subsidizes skua reproduction at this site, which could affect the food web of this low-productivity ecosystem.     

Intraspecific divergence and evolution of a life‐history trade‐off along a successional gradient in Hawaii's Metrosideros polymorpha

The importance of environmental gradients in the diversification of long‐lived tree species is poorly understood. Two morphologically distinct varieties of the endemic Hawaiian tree, 'ōhi'a lehua (Metrosideros polymorpha), are the canopy dominants at alternate extremes of a successional gradient formed by the recurring disturbance of lava flows on east Hawai'i Island. The maintenance of these varieties despite hybridization may be due to disruptive selection at either end of the successional gradient. To test this hypothesis, seeds from three, replicate monotypic stands of each variety on east Hawai'i Island were germinated and the resulting seedlings grown under four combinations of light and nitrogen levels in a greenhouse, and at early‐ and late‐successional field sites. Growth and survivorship measures revealed differential fitness of these varieties in high‐ and low‐light environments in the greenhouse with corresponding differential fitness in early‐ and late‐successional field sites. Unique light‐by‐nitrogen interaction effects on growth were observed in each variety, and only the late‐successional variety appeared to be nitrogen limited. These two varieties exhibit the classic plant life‐history trade‐off between fast growth in high light and high survivorship in shade, but notably within a single tree species. These findings strongly implicate a role for Hawaii's striking environmental heterogeneity in the emergence of at least two endemic forms of this woody genus.