Population health risk due to dietary intake of toxic heavy metals from Spinacia oleracea harvested from soils collected in and around Tshwane,South Africa

In order to assess the possible health risk associated with the consumption of vegetables harvested from waste dump sites, trace metal levels in Spinacia oleracea planted in soils collected from waste dump sites were investigated. Soil samples from different waste dump sites and mining areas were collected and placed in different pots. Seedlings of S. oleracea were introduced into the pots, harvested after 3 months and analysed for trace metal contents using ICP-MS. From the leaves of the plants, the concentration of Fe was found to be significantly higher than all other trace metals (p < 0.05). The trend in trace metal accumulation from the leaves was in the order Fe > Mn > Zn > Pb > Cu > Cr > Ni > Cd. A significantly different concentration of trace metals in the plant was noticed from different soils in different pots used (p < 0.05). Trace metal concentration from plant parts showed roots > leaves > stem. The risk to human health indicated as Hazard Quotient (HQ) was highest for Zn followed by Cu from all the plant parts. The HQ result showed that humans might be at risk if they consume spinach from these waste dump sites. From the study it was concluded that harvesting/consuming spinach from soil around a waste dump site may be extremely dangerous.     

Root health evaluation of three alfalfa varieties in Kerquin sandy land

Without a robust and healthy root system, establishment, productivity, and persistence are compromised. Consequently, research on alfalfa root morphology and health is very important in development of technology for efficient improvement and production of alfalfa. The objectives of this study were to evaluate the root morphology and health of three alfalfa varieties, Algonquin, Golden Queen, and Yellow Flower and to determine relationships among root morphology traits and root health. Yields from these varieties ranged from 5.83 to 43.93 t/ha, total root length ranged from 215.17 to 708.89 mm, root surface area from 124.95 to 468.37 cm², volume from 3.24 to 57.72 cm³, and forks from 1.25 × 10³ to 10.54 × 10³, and tips from 0.65 × 10³ to 3.17 × 10³. Root infestation score was negatively correlated with yield (r = ?0.997, P < 0.01), and was positively correlated with all root morphology traits (r = 0.466–0.997, P < 0.01), and yield was negatively associated with root morphology traits (r = ?0.755 to ?0.998, p < 0.01) with the exception of root tips (r = 0.448, P < 0.01). Results from these analyses indicated that root infestation score was the lowest averaged over age of alfalfa stand in Algonquin. Yield in 2-year old stands was greater in Golden Queen compared to the other two cultivars.     

Prefermentation to overcome nutrient limitations in food processing wastewater: Comparison of pilot- and bench-scale systems

A bench- and a pilot-scale anaerobic/aerobic system were evaluated for the treatment of high strength tomato-processing wastewater. The pilot-scale anaerobic tank achieved better prefermentation of organic carbon and nitrogen than the bench-scale system, although overall system performance was comparable with more than 99% SBOD removal and 97% SCOD removal. Hydraulic retention time (HRT) and temperature effects were studied in the bench-scale system. Increase of anaerobic HRT from 0.25 day to 0.5 day favored prefermentation and a better effluent quality was achieved, as demonstrated by reduction in TSS concentrations from 66 mg/L to 24 mg/L, SCOD from 103 mg/L to 78 mg/L and SBOD from 8 mg/L to 6 mg/L, respectively. Specific oxygen uptake rate (SOUR) increased from 0.15–0.23 mg O₂/mg VSS day at 25 °C to 0.67–1.24 mg O₂/mg VSS day at 32 °C. Settling characteristics deteriorated from sludge volume index (SVI) of 24–131 mL/g at 25 °C to 115–173 mL/g at 32 °C. Sludge yield decreased from 0.14 g VSS/g COD at 25 °C to 0.098 g VSS/g COD at 32 °C.     

Ear fibroblasts derived from Taiwan yellow cattle are more heat resistant than those from Holstein cattle

The objective of this study was to compare the thermotolerances of ear fibroblasts derived from Holstein (H) and Taiwan yellow cattle (Y) and their apoptosis-related protein expressions with (1, 3, 6, 12, and 24 h) or without heat shock treatment. The results showed that the vaginal temperatures of Y (38.4–38.5 °C) were (P<0.05) lower than that of H (38.8 °C) during the hot season. The apoptotic rates of ear fibroblasts derived from Y (6 h: 1.1%; 12 h: 1.6%; 24 h: 2.6%) were lower (P<0.05) than those of cells derived from H (6 h: 1.8%; 12 h: 4.0%; 24 h: 6.9%), respectively, after heat shock (42 °C). The expression level of apoptosis inducing factor (AIF) in ear fibroblasts derived from H was higher (P<0.05) than those derived from Y after the heat shock treatment for 6 h and 12 h, respectively. The level of cytochrome c of ear fibroblasts derived from H was higher (P<0.05) than those derived from Y after the heat shock treatment for 1–12 h, respectively. The abundances of Caspase-3, Caspase-8 and Caspase-9 of ear fibroblasts derived from H were higher (P<0.05) than those of cells derived from Y after 12 h and 24 h of heat shock, respectively; the Bcl-2/Bax ratios of ear fibroblasts derived from H were lower (P<0.05) than those from Y-derived fibroblasts after heated for 1–24 h. The expression level of HSP-70 of Y-derived ear fibroblasts was also higher (P<0.05) than that from H after the same duration of heat shock treatments. Taken together, the thermotolerance of ear fibroblasts derived from Taiwan yellow cattle was better than that of cells derived from Holstein cattle.     

Investigation of porcine circovirus contamination in human vaccines

DNA from porcine circovirus type 1 (PCV1) and 2 (PCV2) has recently been detected in two vaccines against rotaviral gastroenteritis from manufacturers A and B. We investigated if PCV1 sequences are present in other viral vaccines. We screened seeds, bulks and final vaccine preparations from ten manufacturers using qRT-PCR. We detected 3.8 × 10³ to 1.9 × 10⁷ PCV1 DNA copies/milliliter in live poliovirus seeds for inactivated polio vaccine (IPV) from manufacturer A, however, following inactivation and purification, the finished IPV was PCV1-negative. PCV1 DNA was not detectable in live polio preparations from other vaccine producers. There was no detectable PCV1 DNA in the measles, mumps, rubella and influenza vaccines analysed including material supplied by manufacturer A. We confirmed that the PCV1 genome in the rotavirus vaccine from manufacturer A is near full-length. It contains two mutations in the PCV cap gene, which may result from viral adaptation to Vero cells. Bulks of this vaccine contained 9.8 × 10¹⁰ to 1.8 × 10¹¹ PCV1 DNA copies/millilitre and between 4.1 × 10⁷ and 5.5 × 10⁸ DNA copies were in the final doses. We found traces of PCV1 and PCV2 DNA in the rotavirus vaccine from manufacturer B. This highlights the issue of vaccine contamination and may impact on vaccine quality control.     

Toward scrubbing the bay: Nutrient removal using small algal turf scrubbers on Chesapeake Bay tributaries

Restoration of the Chesapeake Bay poses significant challenges because of increasing population pressure, conversion of farmland to urban/suburban development, and the expense of infrastructure needed to achieve significant and sustained nutrient reductions from agricultural and urban sources. One radical approach for removing non-point source nutrients before they reach the bay is to deploy large-scale algal turf scrubbers along its tributaries. The objective of this study was to determine rates of nutrient removal and algal fatty acid production using small ATS units located along three Chesapeake Bay rivers. Small-scale ATS units (each containing 1 m² growing area) were operated for 5–10 months from April 2007 to April 2008 on three western shore tributaries of the Chesapeake Bay in Maryland: the Bush River, the Patapsco River and the Patuxent River. Total nitrogen (TN) and total phosphorus (TP) removal rates at the Patuxent site fluctuated considerably but averaged 250 mg TN, 45 mg TP m^?2 day^?1 from May to October 2007, then decreased to 16 mg TN, 3 mg TP m^?2 day^?1 from December 2007 to February 2008. Nutrient removal rates at the Bush river site also fluctuated but averaged only 85 mg TN, 10 mg TP m^?2 day^?1 from May to June 2007, before decreasing to <10 mg TN, <1 mg TP m^?2 day^?1 from July to September 2007. The Patapsco River unit began operation in August 2007, reached its maximum removal values of 150 mg TN, 18 mg TP m^?2 day^?1 from mid-October to late-November 2007, then decreased to values of 45 mg TN, 4 mg TP m^?2 day^?1 from November 15, 2007 to mid-April 2008. In the best case (Patuxent site from May to October 2007), daily removal rates of 250 mg N and 45 mg P m^?2 are equivalent to removal rates of 380 kg N and 70 kg P ha^?1 over a 150-day season in Maryland. Fatty acid (FA) content of the harvested material was consistently low (0.3–0.6% of dry weight) and varied little between sites. Mean algal FA production rates (23–54 mg FA m^?2 day^?1) are equivalent to rates of 34–81 kg FA ha^?1 year^?1 based on a 150-day operational season in Maryland.     

Mensurations échographiques de la thyroïde chez des enfants de zéro à 15 ans de la ville de Ouagadougou

Among the methods used to carry out measurements of thyroid, echography is the best because it is noninvasive and more specific. The goals of this study were to determine the normal thyroid volume of the Burkinabe child; to study the parameters which induce a variation of thyroid volume and to compare our values with those from other localities. Through a cross-sectional study, 240 children from the town of Ouagadougou benefited from a thyroid echography. Thus the height (h), the transverse diameter (dt) and the thickness (e) were measured. Volume (vl) of each lobe was estimated by: vl = h × dt × e × 3.14/6. The total volume (VT) is the sum of the volumes of each lobe, the isthmus being neglected. A correlation set at the threshold p < 0.001 has been carried out between VT and some anthropometric parameters. The following averages were observed: before one year, the average volume is of 0.51 L ± 0.21; from one to three years, it is of 0.95 mL ± 0.33; from four to six years, the average volume is of 1.35 mL ± 0.43; from seven to nine years, it is of 1.95 mL ± 0.62. From 10 to 12 years, the average volume is of 3.01 mL ± 1.08 and it is of 5.32 mL ± 2.07 from 13 to 15 years. It stands out that the VT is positively associated with the age (r = 0.82), with the size (r = 0.73), with the weight (r = 0.81) and with body surface (r = 0.78). Moreover, the VT is higher among boys than in girls beyond one year of age. This parameter varies according to the area of origin. Lastly, compared with the American, European and Asian averages, our values are smaller. This study enabled us to establish our own standards which we will be able from now on to use in daily practice. However, multicentric studies associated with thyroid and sexual hormones blood levels measurements must be undertaken to ascertain these data.     

Saxitoxin monitoring in three species of Florida puffer fish

Beginning in April 2002, three species of Florida puffer fish from around the state of Florida, USA were monitored for the presence of saxitoxin (STX). In total, 873 southern (Sphoeroides nephelus), 171 checkered (S. testudineus), and 53 bandtail (S. spengleri) puffer fish were collected between 2002 and 2006 from eight regions: Jacksonville, the Indian River Lagoon, Tequesta, the Florida Keys, Charlotte Harbor, Tampa Bay, Cedar Key, and Apalachicola. Emphasis was placed on collecting specimens from the Indian River Lagoon (IRL), where recreational harvesting of puffer fish led to 28 cases of saxitoxin puffer fish poisoning (SPFP) between January 2002 and May 2004. Southern puffer fish from the northern IRL routinely contained the highest concentrations of STX, with average levels in the skin of 1787 μg STXequiv./100 g tissue. Elevated concentrations were also found in the muscle (1102 μg STXequiv./100 g), gut contents (539 μg STXequiv./100 g), gonads (654 μg STXequiv./100 g), and liver (214 μg STXequiv./100 g). Lower, yet significant (above the action limit of 80 μg STXequiv./100 g tissue), concentrations of STX were also detected in the skin (599 μg STXequiv./100 g), muscle (233 μg STXequiv./100 g), gut contents (197 μg STXequiv./100 g), and gonads (239 μg STXequiv./100 g) of southern puffer fish from Tequesta in the southern IRL, as well as in the gonads (122 μg STXequiv./100 g) of Jacksonville southern puffer fish and the skin (265 μg STXequiv./100 g) of Tampa Bay southern puffer fish. STX concentrations above the action limit were also found in the skin of bandtail puffer fish from the IRL (620 μg STXequiv./100 g), Tequesta (374 μg STXequiv./100 g), and the Florida Keys (230 μg STXequiv./100 g). Checkered puffer fish collected from the IRL, Tequesta, and the Florida Keys on average were nontoxic, containing STX levels below the action limit in all tissues.     

Skeletal muscle pericyte subtypes differ in their differentiation potential

Neural progenitor cells have been proposed as a therapy for central nervous system disorders, including neurodegenerative diseases and trauma injuries, however their accessibility is a major limitation. We recently isolated Tuj1 + cells from skeletal muscle culture of Nestin–GFP transgenic mice however whether they form functional neurons in the brain is not yet known. Additionally, their isolation from nontransgenic species and identification of their ancestors is unknown. This gap of knowledge precludes us from studying their role as a valuable alternative to neural progenitors. Here, we identified two pericyte subtypes, type-1 and type-2, using a double transgenic Nestin–GFP/NG2–DsRed mouse and demonstrated that Nestin–GFP +/Tuj1 + cells derive from type-2 Nestin–GFP +/NG2–DsRed +/CD146 + pericytes located in the skeletal muscle interstitium. These cells are bipotential as they generate either Tuj1 + cells when cultured with muscle cells or become “classical” α-SMA + pericytes when cultured alone. In contrast, type-1 Nestin–GFP ?/NG2–DsRed +/CD146 + pericytes generate α-SMA + pericytes but not Tuj1 + cells. Interestingly, type-2 pericyte derived Tuj1 + cells retain some pericytic markers (CD146 +/PDGFRβ +/NG2 +). Given the potential application of Nestin–GFP +/NG2–DsRed +/Tuj1 + cells for cell therapy, we found a surface marker, the nerve growth factor receptor, which is expressed exclusively in these cells and can be used to identify and isolate them from mixed cell populations in nontransgenic species for clinical purposes.     

Comparison of the lipid properties of captive,healthy wild,and pansteatitis-affected wild Nile crocodiles (Crocodylus niloticus)

The results presented describe and compare the fatty acid composition and melting properties of captive, healthy wild, and pansteatitis-affected wild crocodiles (Crocodylus niloticus). Differences in fatty acid composition between intramuscular and adipose fat is noted in captive crocodiles, and the latter differs from wild crocodiles as a result of different diets. Adipose fat of healthy wild crocodiles differs minimally from diseased ones, respectively with 37.3 ± 2.6% vs. 43.2 ± 2.3% monounsaturated fatty acids, and 43.2 ± 2.9% in dead crocodiles, while polyunsaturated fatty acids decrease from 27.3 ± 1.9% to as low as 21.9 ± 3.6% respectively. Of the unsaturated fatty acids 18:2n? 6 decreased from 6.5 ± 2.6% in unaffected crocodiles to 3.5 ± 0.6% in highly affected and 3.2 ± 0.4% in dead crocodiles, and 22:5n?3 from 2.8 ± 0.6% to 1.8 ± 0.3% and 2.2 ± 0.3% respectively. The melting properties as determined by differential scanning calorimetry show that extracted adipose fat is a small degree softer in pansteatitis-affected tissue, specifically in the temperature range 7–36 °C, and does not contribute to the hard texture noted for adipose fat tissue of pansteatitis-affected animals. A high moisture content of 51.0 ± 19.7% of the fat tissue of pansteatitis-affected animals vs.17.1 ± 8.0% of healthy ones, suggests that physiological changes due to interstitial inflammation may contribute to the hard texture.     

Removal of iron,arsenic and coliform bacteria from water by novel constructed soil filter system

The present paper presents results of the study in removal of iron, arsenic and total coliform from drinking water using single-pass constructed soil filter (CSF). Results indicated that arsenic levels ranged from 0.5 to less than 10 μg l^?1 levels; iron from 5 to less than 0.3 mg l^?1 and coliform from 10^?5 to less than 5 CFU/100 ml. The results revealed very high removal efficiency, i.e., over 99% and water quality as per WHO standard.     

Designing better water troughs: 2. Surface area and height,but not depth,influence dairy cows’ preference

Water consumption affects milk production of dairy cows. In a previous study, we found that dairy cows preferred to drink from larger than from smaller troughs and that intake was higher when water was offered in the larger, preferred troughs. In this study, we investigated some of the trough's characteristics that may underlie such preference. The volume of water consumed, time spent drinking and number of sips taken by cows (n = 18) were compared when water was offered in two troughs differing in surface area (1.13 m² or 0.28 m²; experiment 1), height (30 cm or 60 cm; experiment 2) or depth (30 cm or 60 cm; experiment 3). In each experiment, each cow was tested individually for six consecutive days with the troughs randomly placed in each side. In experiment 1, cows took more sips (P < 0.01), spent more time drinking (P < 0.01) and drank more water (P < 0.01) from the trough with larger surface area. In experiment 2, cows took more sips from the higher than from the lower trough (P < 0.02) and showed a tendency to consume more water (P = 0.08) and to spend more time drinking (P = 0.08) from the higher than from the lower trough. Trough depth did not influence any of the variables recorded.     

Inventory and input–output analysis of CO2 emissions by fossil fuel consumption in Beijing 2007

With the most recent statistics available, a concrete emissions inventory is compiled for an input–output analysis to investigate the embodied CO₂ emissions induced by fossil fuel combustion of Beijing economy in 2007. Results show that the total direct CO₂ emissions amount to 9.45E + 07 t, within which 56.81% are released from coal combustion, 11.50% from coke combustion, 9.03% from kerosene combustion, 8.70% from natural gas and 6.40% from diesel, respectively. The average intensity of secondary industries (3.12 t/1E + 4 Yuan) is 0.65 times larger than that of primary industries (1.89 t/1E + 4 Yuan) and 1.58 times larger than that of tertiary industries (1.21 t/1E + 4 Yuan). The sector of Construction Industry contributes the largest share (21.98%) of CO₂ emissions embodied in final demand for Beijing due to its considerable capital investment. Beijing is a net importer of embodied CO₂ emissions with total import and export of 3.06E + 08 and 2.00E + 08 t, respectively. Results of this study provide a sound scientific database for effective policy making in Beijing to reduce CO₂ emissions.     

Temperature-dependent development models of Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae) Q biotype on three host plants

Temperature-dependent development of the sweet potato whitefly, Bemisia tabaci (Gennadius), Q biotype was examined on three host plants (bell pepper, oriental melon, and eggplant) at nine temperatures (15, 17.5, 20, 22.5, 25, 27.5, 30, 32.5, and 35 °C). Egg development time (least squares [LS]-mean ± LS-standard error [SE]) varied from 31.78 ± 0.29 days at 15 °C to 4.93 ± 0.25 days at 32.5 °C on bell pepper, from 21.27 ± 0.20 days at 17.5 °C to 4.02 ± 0.23 days at 32.5 °C on oriental melon, and from 26.92 ± 0.19 days at 15 °C to 5.14 ± 0.18 days at 30 °C on eggplant. Nymph development time (LS-mean ± LS-SE) varied from 76.54 ± 0.96 days at 15 °C to 12.96 ± 0.68 days at 27.5 °C on bell pepper, from 48.78 ± 0.38 days at 17.5 °C to 11.32 ± 0.38 days at 32.5 °C on oriental melon, and from 73.08 ± 1.23 days at 15 °C to 11.89 ± 0.70 days at 27.5 °C on eggplant. A non-linear relationship between developmental rate and temperature was described by the Taylor model, and developmental variation was described by the two-parameter Weibull function.     

Bioavailability and extraction of heavy metals from contaminated soil by Atriplex halimus

Pot experiments were performed to evaluate the phytoremediation capacity of plants of Atriplex halimus grown in contaminated mine soils and to investigate the effects of organic amendments on the metal bioavailability and uptake of these metals by plants. Soil samples collected from abandoned mine sites north of Madrid (Spain) were mixed with 0, 30 and 60 Mg ha⁻¹ of two organic amendments, with different pH and nutrients content: pine-bark compost and horse- and sheep-manure compost. The increasing soil organic matter content and pH by the application of manure amendment reduced metal bioavailability in soil stabilising them. The proportion of Cu in the most bioavailable fractions (sum of the water-soluble, exchangeable, acid-soluble and Fe–Mn oxides fractions) decreased with the addition of 60 Mg ha⁻¹ of manure from 62% to 52% in one of the soils studied and from 50% to 30% in the other. This amendment also reduced Zn proportion in water-soluble and exchangeable fractions from 17% to 13% in one of the soils. Manure decreased metal concentrations in shoots of A. halimus, from 97 to 35 mg kg⁻¹ of Cu, from 211 to 98 mg kg⁻¹ of Zn and from 1.4 to 0.6 mg kg⁻¹ of Cd. In these treatments there was a higher plant growth due to the lower metal toxicity and the improvement of nutrients content in soil. This higher growth resulted in a higher total metal accumulation in plant biomass and therefore in a greater amount of metals removed from soil, so manure could be useful for phytoextraction purposes. This amendment increased metal accumulation in shoots from 37 to 138 mg pot⁻¹ of Cu, from 299 to 445 mg pot⁻¹ of Zn and from 1.8 to 3.7 mg pot⁻¹ of Cd. Pine bark amendment did not significantly alter metal availability and its uptake by plants. Plants of A. halimus managed to reduce total Zn concentration in one of the soils from 146 to 130 mg kg⁻¹, but its phytoextraction capacity was insufficient to remediate contaminated soils in the short-to-medium term. However, A. halimus could be, in combination with manure amendment, appropriate for the phytostabilization of metals in mine soils.     

The assessment of anti-tumoral activity of polysaccharide extracted from terrestrial filamentous fungus

Fungal polysaccharides are well-known for the medicinal properties such as antitumor and immunomodulating effects. Hence, this study evaluated antitumor effects of polysaccharide extracted from Fusarium sp. isolated from soil samples of Karaj district, Alborz, Iran along with its taxonomic study. The filamentous fungus strain FK1 was isolated from the soil sample of Karaj, Iran. The strain was identified based on cultural, morphological and 18 S rRNA gene parameters as Fusarium. Further, the strain Fusarium was cultured in fermented broth of modified (PDB) for 10  days at 25 °C. The polysaccharide of strain FK1 was extracted from the mycelium free supernatant by boiling water method and evaluated for antitoxicity effect on two human cancer cell lines: HeLa cell line and Lymphoblastoid cell line (LCL) by MTT method. Findings revealed that water-extracted from mycelia polysaccharide of strain FK1 had the highest cytotoxicity effect against LCL which is the cause of B lymphocyte cancer, at 50  μg/ml concentration dose (114 ± 1.63) followed by 100  μg/ml (105 ± 0.57) and 10  μg/ml (104 ± 0.57), while it did not have a considerable effect on HeLa cell line. Fusarium could be alternative sources as an antitumor component.     

Free-ranging domestic cats reduce the effective protected area of a Polish national park

Poland's Animal Protection Act, as of 2002, made it legal to shoot free-ranging cats and dogs. The act triggered substantial social debate with opponents arguing that this legislation was weakly supported by scientific evidence of the ecological impacts of free-ranging pets. Our main research goal was to examine the activity of free-ranging domestic cats within a Polish protected area by applying radio-telemetry methods to determine space use and degree of encroachment into the national park. We trapped and radio-tracked 19 animals from three sites (focal households) located in Ojcow National Park (ONP) in southern Poland from June 2003 to March 2006. Annual 100% MCP home range size varied from 0.02 km² to 1.46 km², and was significantly larger for males (mean ± SE = 0.79 ± 0.34 km²; median = 0.53 km²) than for females (mean ± SE = 0.13 ± 0.05 km²; median = 0.13 km²). The distance travelled by individual cats from focal sites did not significantly differ between males (mean ± SE = 232.00 ± 21.05 m; median = 191 m) and females (mean ± SE = 232.50 ± 12.47 m; median = 228 m), with maximum distances of 1.5 km for males and 1.1 km for females. All monitored cats were in close proximity to nature reserves and ranged into protected areas without any human control. Cats living in the households in the park and its surrounding buffer zone, roaming at 200 m and 1000 m radius distances from their households, occupied from 6% to 100% of the park area, respectively. Our results reveal that free-ranging domestic cats roam through and potentially impact the entire national park, thus reducing its effective protected area.     

Use of manure concentrations of ash or specific minerals and nitrogen to estimate loss of volatile nitrogen from manure incubated under laboratory conditions

Changes in the ratio of volatile N to non-volatile specific minerals or total ash in manure can potentially be used to estimate losses of N from manure. The aim of this study was to evaluate the accuracy of using specific minerals (P, K, Ca, Mg, and Na) or total ash as markers to estimate volatile N losses from incubated manure slurries. Holstein cows were fed diets with the forage as predominantly corn silage (CS) or alfalfa silage (AS) or a diet identical to the CS diet except 5 g/kg urea was added (CSU). Total output of urine and feces were measured and used to prepare two types of slurry mixtures (all slurry mixtures contained 1200 g of manure per tray): (1) as excreted (AsEx) and (2) AsEx + sand (SAND). The SAND slurry was designed to mimic manure from barns that used sand as a bedding material. Feces and urine were mixed in the same proportion as excreted by the cows for both slurry types and 240 g of sand were added to the SAND slurries. Initial and final (3 d incubation period) slurry weights and concentrations of N, ash, and specific minerals were measured to calculate loss of volatile N. Losses of volatile N from the slurries were estimated as: (N intake ? N milk) ? [N/ash × (ash intake ? ash milk)], where N and ash intake = g/d consumed by the cow; N and ash milk = g/d secreted in milk; and N/ash = ratio of N and ash concentrations in slurry samples at 3 d. To evaluate specific minerals as markers, the same equation was used except that Ca, K, Mg, Na or P replaced the ash term. Measured N losses from the AsEx slurries were 0.44, 0.62, and 1.03 g/3 d for the AS, CS, and CSU treatments and estimated N losses (using N/ash) were 0.29, 0.49, and 1.14 g/3 d. Estimated losses were less than measured losses for cows with negative N balance and greater than measured losses for cows in positive N balance. For AsEx slurries, the use of N to specific mineral ratios was generally less accurate than using the N/ash ratio. Estimated N losses from SAND slurries were extremely inaccurate for all markers. The use of the N/ash ratio to estimate N volatilization from manure shows promise but markers that are in appreciable concentrations in bedding material will not be accurate.     

Direct ethanol production from N-acetylglucosamine and chitin substrates by Mucor species

Chitin, which is a polymer of β-(1–4) linked N-acetyl-d-glucosamine (GlcNAc) residues, is one of the most abundant renewable resources in nature, after cellulose. In this study, we found some native Mucor strains, which can use GlcNAc and chitin substrates as carbon sources for growth and ethanol production. One of these strains, M. circinelloides NBRC 6746 produced 18.6 ± 0.6 g/l of ethanol from 50 g/l of GlcNAc after 72 h and the maximum ethanol production rate was 0.75 ± 0.1 g/l/h. Furthermore, M. circinelloides NBRC 4572 produced 6.00 ± 0.22 and 0.46 ± 0.04 g/l of ethanol from 50 g/l of colloidal chitin and chitin powder after 16 and 12 days, respectively. We also found an extracellular chitinolytic enzyme producing strain M. ambiguus NBRC 8092, and successfully improved ethanol productivity of NBRC 4572 from colloidal chitin using crude chitinolytic enzyme derived from NBRC 8092. The ethanol titer reached 9.44 ± 0.10 g/l after 16 days. These results were the first bioethanol production from GlcNAc and chitin substrates by native organisms, and also suggest that these Mucor strains have great potential for the simultaneous saccharification and fermentation (SSF) of chitin biomass.     

Direct bioconversion of d-xylose to 1,2,4-butanetriol in an engineered Escherichia coli

The compound 1,2,4-butanetriol (BT) is a valuable chemical used in the production of plasticizers, polymers, cationic lipids and other medical applications, and is conventionally produced via hydrogenation of malate. In this report, BT is biosynthesized by an engineered Escherichia coli from d-xylose. The pathway: d-xylose → d-xylonate → 2-keto-3-deoxy-d-xylonate → 3,4-dihydroxybutanal → BT, was constructed in E. coli by recruiting a xylose dehydrogenase and a keto acid decarboxylase from Caulobacter crescentus and Pseudomonas putida, respectively. Authentic BT was detected from cultures of the engineered strain. Further improvement on the strain was performed by blocking the native d-xylose and d-xylonate metabolic pathways which involves disruption of xylAB, yjhH and yagE genes in the host chromosome. The final construct produced 0.88 g L⁻¹ BT from 10 g L⁻¹ d-xylose with a molar yield of 12.82%. By far, this is the first report on the direct production of BT from d-xylose by a single microbial host. This may serve as a starting point for further metabolic engineering works to increase the titer of BT toward industrial scale viability.