Isolation and characterization of indigenous soil bacteria for bioaugmentation of PAH contaminated soil of semiarid Patagonia, Argentina

The aim of this work was to isolate PAH degrading-bacteria from contaminated Patagonia soil with the ability to tolerate the usual environmental stresses (oligotrophic and dryness conditions). Two approaches were utilized to obtain PAH-degrading bacteria from the Patagonian soil. With a traditional enrichment approach only the PAH- degrading strain 36 was isolated. Using a direct isolation approach three PAH-degrading strains (1A, 22A and 22B) were isolated. The phylogenetic analysis revealed that all isolates belonged to Sphingomonas genus. The PAH degrading activity and the resistance to stress conditions of the strains were determined and compared with those of the exogenous PAH-degrading Sphingomonas paucimobilis 20006FA. The strains 1A, 22A and 36 were phylogenetically closely related between them and with the strain 20006FA. The strain 22B, that showed a different phylogenetic position, was more resistant to C-starvation and drying conditions than other Patagonian strains. The effect of the inoculation of these strains on phenanthrene-induced mineralization and elimination was studied in Patagonian soil artificially contaminated, at different environmental conditions. The results suggest that strain 22B is the most suitable strain for bioaugmentation in PAH-contaminated soils of Central Patagonia, due to its adaptation to the usual environmental conditions. Our results show the importance of a detailed physiological characterization of isolates for autochthonous bioaugmentation strategies success.     

Monoterpene emissions from three Nothofagus species in Patagonia,Argentina

Abstract

Isoprenoid emissions have key roles in plant biology and plant interactions with the environment. Global emission inventories of isoprenoid emissions still lack information from a large number species, especially from South American vegetation other than the rainforest ecosystem. A study was conducted to identify the basal emission of isoprenoid under field conditions from three Nothofagus species. The three Nothofagus species were characterized as strong monoterpene emitters while the emission of isoprene was undetectable. The two deciduous species, N. pumilio and N. antarctica, had similar photosynthetic parameters, but monoterpene emission rate and, consequently, the fraction of photosynthetic carbon re-emitted in the atmosphere as monoterpenes, were more than three-fold higher in N. pumilio than in N. Antarctica. The evergreen species N. dombeyi showed intermediate values of both monoterpene emission rate and fraction of photosynthetic carbon re-emitted. The monoterpene emission spectrum was very similar among the three Nothofagus species screened, but clearly different from the spectrum reported in other monoterpene-emitting species of the Fagaceae family. The importance of these findings for atmospheric chemistry and phylogenic evolution are discussed.     

Biodiversity of cold-adapted yeasts from glacial meltwater rivers in Patagonia, Argentina

The occurrence of culturable yeasts in glacial meltwater from the Frías, Casta?o Overo and Río Manso glaciers, located on Mount Tronador in the Nahuel Huapi National Park (Northwestern Patagonia, Argentina) is presented. Subsurface water samples were filtered for colony counting and yeast isolation. The total yeast count ranged between 6 and 360 CFU L(-1). Physiologic and molecular methods were employed to identify 86 yeast isolates. In agreement with yeast diversity data from studies for Antarctic and Alpine glaciers, the genera Cryptococcus, Leucosporidiella, Dioszegia, Rhodotorula, Rhodosporidium, Mrakia, Sporobolomyces, Udeniomyces and Candida were found. Cryptococcus and Leucosporidiella accounted for 50% and 20% of the total number of strains, respectively. Among 21 identified yeast species, Cryptococcus sp. 1 and Leucosporidiella fragaria were the most frequent. The typically psychrophilic Mrakia yeast strain and three new yeast species, yet to be described, were also isolated. All yeast strains were able to grow at 5, 10, and 15 degrees C. Among yeast strains expressing extracellular enzymatic activity, higher proteolytic and lipolytic activities were obtained at 4 degrees C than at 20 degrees C.     

Molecular characterization of carotenogenic yeasts from aquatic environments in Patagonia, Argentina

Fifteen aquatic environments (lakes, lagoons and rivers) of glacial origin in the northern Andean Patagonia (Argentina) were surveyed for the occurrence of red yeasts. Subsurface water samples were filtered and used for colony counting and yeast isolation. A preliminary quantitative analysis indicated that total yeast counts ranged between 0 and 250 cells l⁻¹. A polyphasic approach including physiological and molecular methods was used for the identification of 64 carotenogenic yeast strains. The molecular characterisation of the isolates was based on the mini/microsatellite-primed PCR technique (MSP-PCR) employing the (GTG)₅ and the M13 primers. Comparison of representative fingerprints of each group with those of the type strains of pigmented yeasts allowed the expeditious identification of 87.5% isolates. The sequence analysis of the D1/D2 domains of the 26S rDNA was employed to confirm identifications and in the characterization of the unidentified MSP-PCR groups. Teleomorphic yeast species were detected by performing sexual compatibility assays. The isolates corresponded to 6 genera and 15 yeast species, including four new yeast species of the genera Cryptococcus (1), Rhodotorula (1) and Sporobolomyces (2). Rhodotorula mucilaginosa was found in the majority of the samples and represented ca. 50% of the total number of isolates. However, this yeast was not detected in aquatic environments with very low anthropic influence. Other frequent yeast isolates were teleomorphic yeast species of Rhodosporidium babjevae, R. kratochvilovae and Sporidiobolus salmonicolor. This study represents the first report on red yeast occurrence and biodiversity in northwestern Patagonia. This revised version was published online in June 2006 with corrections to the Cover Date.     

Response of bacteria and phytoplankton to contaminated sediments from Trenton Channel,Detroit River

Several types of bioassays were used in 1986 and 1987 to investigate the effect of contaminated sediments on natural populations of bacteria and phytoplankton from the Trenton Channel, Detroit River. The approach included the measurement of uptake of ³H-glucose or ³H-adenine by bacteria and ¹⁴C-bicarbonate by phytoplankton in the presence of different amounts of Trenton Channel and Lake Michigan (control) sediments. Trenton Channel sediments are contaminated by high levels of toxic organic compounds and metals, especially zinc, lead, and copper. Because levels of biomass of bacteria and phytoplankton varied widely among the different bioassays, it was necessary to adjust uptake rates for biomass. Biomass adjustments were made using acridine orange counts for bacteria and chlorophyll measurements for phytoplankton. The results show a statistically significant suppression of uptake of substrates for both bacteria and phytoplankton with increasing amounts of sediment. Uptake was suppressed as much as 90 percent for bacteria and 93 percent for phytoplankton at 1200 mg l^-1 of Trenton Channel sediments compared to bioassays without sediment. Uncontaminated Lake Michigan sediment suppressed uptake much less than Detroit River sediment; the difference in suppression of uptake between the two types of sediment was statistically significant for both bacteria and phytoplankton.Contribution No. 518 of the Center for Great Lakes and Aquatic Sciences of the University of Michigan.     

Dentition and histology in titanosaurian dinosaur embryos from Upper Cretaceous of Patagonia,Argentina

Abstract: Exceptionally preserved sauropod embryos from the Late Cretaceous Anacleto Formation in Auca Mahuevo (Neuquén Province, Argentina) have provided fundamental information on titanosaurian ontogeny. This paper describes the dental composition, disposition and microstructure of the specimens. Embryonic teeth show size disparity, with lengths that vary from 1 to 3 mm and diameters ranging from 0.15 to 0.26 mm, with the most frequent length values between 2.5 and 3 mm. Apparently, a typical ‘pencil‐like’ tooth morphology and a dental formula of Pm 4, M 7–8/D10? remained constant during titanosaurian ontogeny, whereas the arrangement of teeth in the skull shows notable ontogenetic changes. Absence of wear facets on teeth suggests a lack of prenatal chewing movements. The enamel proportion is significantly higher in embryos than in mature titanosaurs, which suggests that this relationship varies during ontogeny. Embryonic bony tissue is composed of highly vascularized, cellular woven bone. The absence of osteonal tissue, the high degree of vascularization, the presence of numerous osteocytes and poor development of periosteal bone reveals that the Auca Mahuevo titanosaurs would have had a high early growth rate and that they were buried at a relatively advanced embryonic stage.     

Diversity and enzyme properties of protease-producing bacteria isolated from sub-Antarctic sediments of Isla de Los Estados, Argentina

Protease-producing bacteria isolated from sub-Antarctic marine sediments of Isla de Los Estados (Argentina) were characterized, and the thermal inactivation kinetics of their extracellular proteases compared. Isolates were affiliated with the genera Pseudoalteromonas, Shewanella, Colwellia, Planococcus, and a strain to the family Flavobacteriaceae. Colwellia strains were moderate psychrophiles (optimal growth at about 15°C, maximum growth temperature at around 25°C). 16S rRNA phylogenetic analysis revealed that these strains and Colwellia aestuarii form a distinct lineage within the genus. The remaining isolates were psychrotolerant and grew optimally between 20 and 25°C; two of them represent potentially novel species or genus (16S rRNA < 97% sequence similarity). The thermostability of the extracellular proteases produced by the isolates was analysed, and the inactivation rate constant (k _in), the activation energy (Ea_in) and the activation Gibbs free energy of thermal inactivation (ΔG ^* _in) determined. ΔG ^* _in, calculated at 30°C, varied between 97 and 124 kJ/mol. Colwellia enzyme extracts presented the highest thermosensitivity, while the most thermostable protease activity was shown by Shewanella spp. These results demonstrated that the stability to temperature of these enzymes varies considerably among the isolates, suggesting important variations in the thermal properties of the proteases that can coexist in this environment.     

Vertebral pneumatic structures in the Early Cretaceous sauropod dinosaur Pilmatueia faundezi from northwestern Patagonia,Argentina

One of the diagnostic characters of dicraeosaurid sauropods is a reduction of pneumatization of dorsal and caudal vertebrae relative to their Flagellicaudata sister taxon, Diplodocidae. Here, we analyse pneumatic structures in the dicraeosaurid sauropod Pilmatueia faundezi, compare them to those of diplodocoids and report the first record of camerate chambers in a dicraeosaurid. The pneumatic structures are in a posterior cervical centrum (MLL-Pv-002) and consist of lateral pneumatic fossae on the centrum that communicate internally with large camerae. By contrast, Pilmatueia's dorsal and caudal vertebrae (MLL-Pv-005-016) lack pneumatic fossae on the centra, which is consistent with the previously reported reduced pneumaticity in dicraeosaurids. Nevertheless, the base of the neural arch and possibly the base of the bifid neural spines of a posterior dorsal vertebra (MLL-Pv-005) show pneumatic internal chambers. The pneumatic features of the Pilmatueia cervical centrum and dorsal neural arch we describe indicate that the degree of pneumatization is variable within dicraeosaurids.     

Heavy metal and other trace elements in native mussel Diplodon chilensis from Nothern Patagonia Lakes, Argentina

Native mussels, Diplodon chilensis, were sampled from four lakes in Nahuel Huapi National Park, Northern Patagonia, Argentina in order to evaluate heavy-metal distribution in the region and to assess the contribution of this compartment of the trophic web to their circulation in the food chain. The concentration of potential pollutants Ag, As, Cr, Hg, Sb, and Se, and other nine elements of interest (Ba, Br, Ca, Co, Cs, Fe, Na, Sr, and Zn) were determined in Diplodon chilensis pooled samples. Digestive glands were analyzed separately from soft tissues. Geological tracers Sc, Ta, Th, and rare earth elements were also determined in order to discriminate lithophile contributions. Elemental concentrations of Ba, Br, Fe, Sr, Se, and Zn in total soft tissues samples do not show significant differences among sampling sites. Arsenic and Cr contents in total soft tissues and digestive gland pooled samples are higher in sampling points close to zones with human settlements. Silver contents in samples collected in Lake Nahuel Huapi were higher than in the other lakes studied, and up to 50-fold higher than the sample collected in Lake Traful, considered as the reference. Mercury highest concentration values measured in total soft tissues pooled samples from lakes Nahuel Huapi and Moreno were found to be similar to those observed in other reported Hg contamination situations, and they are three to five times higher than those of the reference samples collected in Lake Traful.     

Enhanced benthic activity in sandy sublittoral sediments: Evidence from 13C tracer experiments

In situ and on-board pulse-chase experiments were carried out on a sublittoral fine sand in the German Bight (southern North Sea) to investigate the hypothesis that sandy sediments are highly active and have fast turnover rates. To test this hypothesis, we conducted a series of experiments where we investigated the pathway of settling particulate organic carbon through the benthic food web. The diatom Ditylum brightwellii was labelled with the stable carbon isotope ¹³C and injected into incubation chambers. On-board incubations lasted 12, 30 and 132 h, while the in situ experiment was incubated for 32 h. The study revealed a stepwise short-term processing of a phytoplankton bloom settling on a sandy sediment. After the 12 h incubation, the largest fraction of recovered carbon was in the bacteria (62%), but after longer incubation times (30 and 32 h in situ) the macrofauna gained more importance (15 and 48%, respectively), until after 132 h the greatest fraction was mineralized to CO₂ (44%). Our findings show the rapid impact of the benthic sand community on a settling phytoplankton bloom and the great importance of bacteria in the first steps of algal carbon processing.     

Decrease in preferred temperature in response to an immune challenge in lizards from cold environments in Patagonia,Argentina

In ectotherms, the likelihood of surviving an infection is determined by the efficiency of thermoregulation, the availability of a variety of thermal microenvironments, the individual's health status, and the virulence of the infective agent. Physiological and behavioral demands related to an efficient immune response entail a series of costs that compete with other vital activities, specifically energy storage, growth, reproduction, and maintenance functions. Here, we characterize the thermal biology and health status by the presence of injuries, ectoparasites, body condition, and individual immune response capacity (using phytohemagglutinin in a skin-swelling assay) of the southernmost lizards of the world, Liolaemus sarmientoi, endemic to a sub-optimal, cold environment in Patagonia, Argentina. In particular, we study the effect of a bacterial endotoxin (lipopolysaccharide; LPS-treatment) on thermoregulation. We found that the field-active body temperature (T_b) was much lower than the preferred body temperature (T_p) obtained in the laboratory. All the individuals were in good body condition at the beginning of the experiments. The phytohemagglutinin test caused detectable thickening in sole-pads at 2 h and 24 h post-assay in males and non-pregnant females, indicating a significant innate immune response. In the experimental immune challenge, the individuals tended to prefer a low body temperature after LPS-treatment (2 h post-injection) and developed hypothermia, while the control individuals injected with phosphate buffered saline (PBS), maintained their body temperature throughout the trial. In both the LPS-treatment and PBS-control individuals, BC declined during the experiment. Hypothermia may allow this southernmost species to optimize the use of their energetic resources and reduce the costs of thermoregulation in a cold-temperate environment where they rarely attain the mean T_p (35.16 °C) obtained in laboratory.     

Molecular application for identification of polycyclic aromatic hydrocarbons degrading bacteria (PAHD) species isolated from oil polluted soil in Dammam,Saud Arabia

Soil contamination with petroleum hydrocarbon products such as diesel and engine oil is becoming one of the major environmental problems. This study describes hydrocarbons degrading bacteria (PHAD) isolated from long-standing petrol polluted soil from the eastern region, Dammam, Saudi Arabia. The isolated strains were firstly categorized by accessible shape detection, physiological and biochemistry tests. Thereafter, a technique established on the sequence analysis of a 16S rDNA gene was used. Isolation of DNA from the bacterial strains was performed, on which the PCR reaction was carried out. Strains were identified based on 16S rDNA sequence analysis, As follows amplified samples were spontaneously sequenced automatically and the attained results were matched to open databases. Among the isolated bacterial strains, S1 was identified as Staphylococcus aureus and strain S1 as Corynebacterium amycolatum.     

PAHs,OCPs and PCBs in sediments from three catchments in Durban,South Africa

Polycyclic aromatic hydrocarbons (PAHs), organochlorine pesticides (OCPs) and polychlorinated biphenyls (PCBs) were analysed from sediment samples collected in 2012 from rivers, surface runoff canals and estuaries in three highly urbanised catchments in durban, KwaZulu-Natal, South Africa. PAHs were ubiquitous, at concentrations between 36–6 800 ng g^?1 dry mass (dm). Congener ratio diagnosis suggests the PAHs were derived predominantly from pyrogenic sources. Four OCPs and/or their metabolites were detected at varying frequencies and concentrations. Of these, dichlorodiphenyltrichloroethane (ddT) and metabolites were most frequently detected and were at a high concentration in sediment in some systems. Toxaphene was detected at a high concentration at some sites. The total PCB concentration varied widely, from below the method detection limit to 110 ng g^?1 (dm). Based on the comparison of chemical concentrations to international sediment quality guidelines, PAH, OCP and PCB concentrations in most sediment samples posed a low risk to sediment-dwelling organisms. However, in some instances the risk was moderate or high. It is recommended that these compounds be monitored more frequently and comprehensively in aquatic ecosystems to better understand the ecological and human health implications.     

Butyltins in sediments and biota from the Pearl River Delta,South China

Abstract

Butyltins in sediments and biota collected from the Pearl River Delta, South China were analyzed by a gas chromatograph–atomic emission detector (GC-AED). The concentrations of tributyltin (TBT) in the sediments ranged from 1.7 to 379.7 ng/g dry weight. Their spatial distribution suggested that shipping activities, especially shipyards were mostly responsible for the TBT contamination in the region. A good linear relationship was observed between the DBT (dibutyltin)/TBT and MBT (monobutyltin) /TBT ratios of the samples in the Pearl River, the Pearl River estuary and the West River, suggesting that TBT in these areas came from similar sources but had undergone a different extent of degradation. The butyltins in the Macao waters seemed to have undergone a different degradation process. All TBT concentrations of the fish, mussel and shrimp samples collected in this study were below the seafood tolerable average residue level (TARL).     

Microeukaryotic diversity in marine environments,an analysis of surface layer sediments from the East Sea

Molecular techniques, based on clone library of 18S rRNA gene, were employed to ascertain the diversity of microeukaryotic organisms in sediments from the East Sea. A total of 261 clones were recovered from surface sediments. Most of the clone sequences (90%) were affiliated with protists, dominated by Ciliates (18%) and Dinoflagellates (19%) of Alveolates, phototrophic Stramenopiles (11%), and Cercozoa (20%). Many of the clones were related to uncultivated eukaryotes clones retrieved from anoxic environments with several highly divergent 18S rRNA gene sequences. However, no clones were related to cultivated obligate anaerobic protists. Protistan communities between subsurface layers of 1 and 9 cm shared 23% of total phylotypes which comprised 64% of total clones retrieved. Analysis of diversity indices and rarefaction curve showed that the protistan community within the 1 cm layer exhibited higher diversity than the 9 cm layer. Our results imply that diverse protists remain to be uncovered within marine benthic environments.     

Geochemical distribution of heavy metals in sediments from sewage fed fish ponds from Kolkata Wetlands,India

Abstract

Kolkata wetlands are the largest sewage fed wetlands in the world. They have been used for aquaculture since 1960. Geochemical distribution of heavy metals (Cr, Cu, Mn, Fe, Zn, Pb, Ni and Al) has been studied in surface sediments using single and sequential extractions techniques. The metal concentrations in sediments were in the following order: Fe>Al> Mn>Zn>Cu> Pb>Cr> Ni, and the average concentrations were 29 μg g^?1, 54 μg g^?1, 328 μg g^?1, 32747 μg g^?1, 169 μg g^?1, 38 μg g^?1, 25 μg g^?1 and 23371 μg g^?1 dry weights for Cr, Cu, Mn, Fe, Zn, Pb, Ni and Al, respectively. Water-soluble percentages of the trace elements are quite low (<0.01–3.75%) but in the presence of chelating agents in the sediments increase the bioavailability of trace elements (2–58%). About 40% of trace elements are in the stable form as a residual fraction of the sediment and more than 50% (nonresidual fraction) metal contamination of the Kolkata wetland sediments were from anthropogenic inputs. The contamination risks of Cr, Mn, Zn, Pb, and Ni are high as their potential availabilities are 70.96%, 58.01%, 63.13%, 55.62%, and 52.15% respectively. The mean concentration of most of the heavy metals in sediments does not exceed the recommended reference values. Zinc and lead concentrations were greater than background level and Interim Sediment Quality Guidelines but lower than Probable Effect Level. Therefore a regular program for monitoring the distribution of heavy metals in water, sediments and biota should be imposed on sewage fed fish ponds of the Kolkata wetland ecosystem.     

Impact of incubation period on biodegradation of petroleum hydrocarbons from refinery wastewater in Kuantan,Malaysia by indigenous bacteria

This work studied the biodegradation of petroleum hydrocarbons (PHCs) extracted from refinery wastewater to produce industrially important by-products at different incubation periods. Two out of 13 bacterial isolates, KRD2 and KRA4 were isolated. Dichloromethane was used to extract the PHC, and gas chromatography-mass spectrometry (GC-MS) analysis revealed that the refinery wastewater PHC was successfully biodegraded using the selected bacterial isolates within 15 days of incubation. Both KRD2 and KRA4 isolates degraded all 13 initially extracted PHC compounds within 5 days, except C13BD and C9BD, which produced 6 and 4 compounds as secondary metabolites with peak area percentages of 1.58, 1.38, 0.85, 29.94, 7.59, and 11.16% and 3.55, 2.88, 52.31, and 6.14%, respectively. These metabolites have been reported in industrial and medical applications. After 10 days, only 6 and 8 compounds were degraded by both isolates, respectively, and C11PAD compound was produced, as well as C5PAD, C7PAD, and C13PAD. After 15 days, it was clear that all the initial PHC compounds have been completely degraded by both isolates. Metabolites C5PAD, C6PAD, C8PAD, and C13PAD were produced by KRD2, and metabolites C5PAD, C6PAD, C8PAD, and C9PAD were produced by KRA4 at different peak areas. The alignment revealed that the KRA4 isolate was included in the genus Chryseobacterium gambrini, while KRD2 isolate was successfully identified as Mycobacterium confluentis using the Biolog microbial identification system. The incubation period evidently affected biodegradation process by indigenous degraders. These effective bacteria were shown to be of great potential for further application in biodegradation technology of PHC contaminated refinery wastewater to produce industrially important by-products.     

Evidence for salt diffusion from sediments contributing to increasing salinity in the Salton Sea, California

Geochemical investigations of interstitial waters from the Salton Sea, CA reveal evidence of concentrated brines in the sediments underlying the lakes two basins. The brines are likely caused by the gradual dissolution of evaporite deposits. The chemical composition of the brine in the northern basin is dominated by magnesium and sulfate and differs from the southern basin where the dominant components are sodium and chloride. Sediment depth distributions of major ions and porosity indicate diffusion of salts from the sediments into the overlying waters in both basins. Benthic fluxes have been calculated for the four most abundant ions: magnesium, sodium, sulfate and chloride. For the northern basin we calculate diffusive fluxes of 3.7 × 10^–2, 8.2 × 10^–2, 44 × 10^–2, and 5.4 × 10^–2 g cm^–2 yr^–1 for magnesium, sodium, sulfate and chloride, respectively. For the southern basin we calculate diffusive fluxes of 0.9 × 10^–2, 9.7 × 10^–2, 6.9 × 10^–2, and 25 × 10^–2 g cm^–2 yr^–1 for these same ions. By scaling up our results we estimate the salinity flux from the sediment to the water column to be between 3.6 × 10⁴ and 3.6 × 10⁵metric tons per year, equivalent to 1–10% of the riverine input. These results are important for developing strategies to combat rising salinity in the Salton Sea, CA.