Examining the Microbial Degradation of Naphthenic Acids Using Stable Isotope Analysis of Carbon and Nitrogen

Naphthenic acids (NAs) are a complex group of naturally occurring oil sands constituents that constitute a significant portion of the dissolved organic carbon (DOC) pool available for microbial degradation in the process-related waste water associated with oil sands mine sites. One approach to understanding the biological fate of oil sands process-derived carbon and nitrogen in aquatic reclamation of the mine sites involves the use of stable isotope analyses. However, for stable isotope analyses to be useful in such field-based assessments, there is a need to determine how microbial degradation of a complex mixture of NAs might change the stable isotope values (δ ¹³C, δ ¹⁵N). In batch cultures and semi-continuous laboratory microcosms, utilization of a commercial mixture of NAs by oil sands-derived microbial cultures resulted in microbial biomass that was similar or slightly ¹³C enriched (1.4‰ to 3.0‰) relative to the DOC source, depending on the length of incubation. Utilization of a NA-containing extract of oil sands processed water resulted in greater ¹³C enrichment of microbial biomass (8.5‰) relative to the DOC source. Overall, the δ ¹³C of the DOC comprised of complex mixtures of NAs showed minimal change (?0.5‰ to ?0.1‰) during the incubation period whereas the δ ¹³C of the dissolved inorganic carbon (DIC) was more variable (?5.0‰ to +5.4‰). In tests where the concentration of available nitrogen was increased, the final biomass values were ¹⁵N enriched (3.8‰ to 8.4‰) relative to the initial biomass. The isotope trends established in this study should enhance our ability to interpret field-based data from sites with hydrocarbon contamination, particularly in terms of carbon source utilization and ¹⁵N enrichment.     

The Impact of Hardwood Line-Planting on Tree and Amphibian Diversity in a Secondary Subtropical Wet Forest of Southeast Puerto Rico

The impact on tree and amphibian diversity of line-planting of tropical hardwoods—mahogany (Swietenia macrophylla × S. mahagoni) and mahoe (Hibiscus elatus)—was studied in a secondary subtropical wet forest of Puerto Rico. Common coqui (Eleutherodactylus coqui) and melodious coqui (E. wightmanae) are the most frequent frog species; forest coqui (E. portoricensis) is less abundant. Although relative abundance means were slightly greater in the undisturbed forest and during the wet season, differences were not statistically significant suggesting that line-planting did not significantly affect amphibian diversity. The line-planted areas had a slightly higher, but not statistically significant diversity, richness, and evenness of tree species than the unplanted forest. Multi-response permutation procedure (MRPP) showed statistically significant community composition differences between line-planting and control plot trees (T = ?5.89, A = .86; p < .001). But mean similarity among plots in both the line-planted and control plots was relatively low at less than 50% of shared species, indicating high diversity of vegetation in the overall forest area. Canopy cover by tree species greater than 3 cm in dbh was much higher in the undisturbed forest but this difference may be reduced as the young line-planted hardwoods mature. Forest enrichment through line-planting of valuable timber species in secondary subtropical wet forest does not significantly affect tree diversity.     

Growing media for horticultural bedding plants yield under agriculture organic systems

The aim of this experiment was to test on-farm growing after composting the mixture of following legume plants Crotalaria juncea, Cajanus cajan, Mucuna aterrima with the grass Sorghum bicolor. The treatments were compared to commercial growing medium (CGM) and CGM+NPK to produce bedding plants of lettuce, broccoli, kale, and cabbage. The nutrient contents in CGM and CGM+NPK were higher than the values found in the mixture of the legumes plants with the grass. The pH was higher in the growing media from legume plants as well as the electrical conductivity in the combination CroSor and CajSor. The water retention capacity was higher in the treatments MucSor and CGM. The chemical attributes indicated that CroSor, CajSor, MucSor and CGM+NPK were better than CGM. The growing media from the mixture of legume with grass are an affordable alternative to produce bedding plants of lettuce, broccoli, kale and cabbage under organic agriculture system.     

Characterization of exopolysaccharides produced by Bifidobacterium longum NB667 and its cholate-resistant derivative strain IPLA B667dCo

Bifidobacteria are natural members of the human intestinal microbiota and some strains are being used as probiotics. Adaptation to bile can allow them to increase survival in gastrointestinal conditions, thus improving their viability. Bifidobacterium longum NB667 and the cholate-resistant strain B. longum IPLA B667dCo produced exopolysaccharides (EPS) that were partially characterized. Analysis by size exclusion chromatography-multiangle laser light scattering indicated that the EPS crude fractions of both strains contained two polymer peaks of different molar mass. On the basis of chromatographic techniques both peaks appeared to be heteropolysaccharides. The smaller peak was mainly composed of glucose, galactose and rhamnose whose molar ratios and linkage types showed slight variations between the EPS fractions of both strains. The bigger peak consisted of glucose and galactose; the monosaccharide composition was identical in the EPS fractions of the two microorganisms, but their infrared spectra presented some differences regarding compounds other than carbohydrates that seem to be associated to the polymer. Differences in the composition of EPS fractions did not affect the capability of crude EPS from B. longum to be fermented by the human intestinal microbiota in fecal batch cultures.     

Diversity of laccase genes from basidiomycetes in a forest soil

Fungal oxidative exo-enzymes lacking substrate specificity play a central role in the cycling of soil organic matter. Due to their broad ecological impact and available knowledge of their gene structure, laccases appeared to be appropriate markers to monitor fungi with this kind of oxidative potential in soils. A degenerate PCR-primer pair Cu1F/Cu2R, specific for basidiomycetes, was designed to assess directly the diversity of laccase genes in soils. PCR amplification of mycelial cultures and fruit-bodies of a wide spectrum of basidiomycetes, covering all functional groups (saprophytes, symbionts, and pathogens), produced multiple DNA fragments around 200 bp. A neighbor-joining tree analysis of the PCR-amplified laccase sequences showed a clear species-specificity, but also revealed that most fungal taxa possess several laccase genes showing a large sequence divergence. This sequence diversity precluded the systematic attribution of amplified laccase of unknown origin to specific taxa. Amplification of laccase sequences from DNA, extracted from a brown (moder) forest soil, showed a specific distribution of laccase genes and of the corresponding fungal species in the various soil horizons (O_h, A_h, B_v). The most organic O_h-horizon displayed the highest gene diversity. Saprophytic fungi appeared to be less widespread through the soil horizons and displayed a higher diversity of laccase genes than the mycorrhizal ones.     

Experimental infection of cats (Felis catus) with Tritrichomonas foetus isolated from cattle

Tritrichomonas foetus is recognized as the causative agent of venereal trichomoniasis in cattle. It is characterized by embryonic and early fetal death and post-coital pyometra, and feline trichomoniasis, manifest as chronic, large bowel diarrhea. Many of the infected cats are less than 2 years old and specific routes of transmission remain unknown. We recently demonstrated that feline isolates of T. foetus can successfully infect heifers, resulting in pathologic changes similar, but not identical to those previously reported as representative of bovine trichomoniasis. In this study, we experimentally infected six cats less than 1 year of age with a bovine (D-1) isolate of T. foetus and one cat with a feline (AUTf-1) isolate of T. foetus. Within 2 weeks, the cat infected with the feline (AUTf-1) isolate was culture positive for trichomonads in weekly fecal samples. At the end of 5 weeks, only one cat infected with the bovine (D-1) isolate was fecal culture positive for trichomonads. At necropsy, the intestine of each cat was removed and divided into five sections (ileum, cecum, anterior, medial and posterior colon). Contents from each section were collected and cultured. The cat infected with the feline (AUTf-1) isolate was culture positive in the ileum, cecum, medial and posterior colon. Two cats infected with the bovine (D-1) isolate were culture positive in the cecum only. Additionally, each intestinal section was submitted to a pathologist for histopathological examination. The combined results indicate that there are demonstrable differences between the feline (AUTf-1) and bovine (D-1) isolates regarding their infectivity in cats.     

Partial Characterization of Buckwheat (Fagopyrum esculentum) Starch

Laboratory-isolated buckwheat (Fagopyrum esculentum) starch was compared to commercial corn and wheat starches. Buckwheat starch granules (2.9–9.3 μm) were round and polygonal with some holes and pits on the surface. Buckwheat starch had higher amylose content, waterbinding capacity, and peak viscosity, and it had lower intrinsic viscosity when compared with corn and wheat starches. Buckwheat starch also showed restricted swelling power at 85–95°C and lower solubility in water at 55–95°C and was more susceptible to acid and enzymatic attack. Gelatinization temperatures, determined by differential scanning calorimetry, were 61.1–80.1°C for buckwheat starch compared to 64.7–79.2°C and 57.1–73.5°C for corn and wheat starches, respectively. A second endotherm observed at 84.5°C was an amylose-lipid complex attributed to the internal lipids in buckwheat starch, as evidenced by selective extraction. The retrogradation of buckwheat, corn, and wheat starch gels was examined after storage at 25, 4, and -12°C for 1–15 days. In general, buckwheat starch retrogradation was slower than that of corn and wheat starch, but it increased as storage time increased, as did that of the other starch pastes. When the values of the three storage temperatures were averaged for each storage period analyzed, buckwheat starch gels showed a lower percentage of retrogradation than did corn and wheat starch gels. Buckwheat starch also had a lower percentage of water syneresis when stored at 4°C for 3–10 days and had better stability to syneresis after three freeze-thaw cycles at -12 and 25°C.