Discovery and directed evolution of a glyphosate tolerance gene

The herbicide glyphosate is effectively detoxified by N-acetylation. We screened a collection of microbial isolates and discovered enzymes exhibiting glyphosate N-acetyltransferase (GAT) activity. Kinetic properties of the discovered enzymes were insufficient to confer glyphosate tolerance to transgenic organisms. Eleven iterations of DNA shuffling improved enzyme efficiency by nearly four orders of magnitude from 0.87 mM-1 min-1 to 8320 mM-1 min-1. From the fifth iteration and beyond, GAT enzymes conferred increasing glyphosate tolerance to Escherichia coli, Arabidopsis, tobacco, and maize. Glyphosate acetylation provides an alternative strategy for supporting glyphosate use on crops.     

Pterocarpan profiles for soybean leaves at different growth stages and investigation of their glycosidase inhibitions

Soybean leaves are eaten as seasonal edible greens in Korea. Analysis of the ethyl acetate extract of these leaves showed that it exhibited potent and selective neuraminidase inhibition, which began at the R3 stage and peaked at R7. Ten pterocarpans, including the new 6a-hydroxypterocarpan 10, were isolated from soybean leaves and their inhibition activities tested against a range of glycosidases. The relationship between structure and enzyme inhibition was investigated: 6a-hydroxypterocarpans exhibited much higher inhibition against neuraminidase (IC(50) = 2.4-89.4 μM) than α-glucosidase (IC(50) = 90.4-?>100 μM). Glyceollin VII (7) displayed 40-fold greater activity (IC(50) = 2.4 μM) against neuraminidase than α-glucosidase (IC(50) = 90.4 μM). On the other hand, coumestanes (1-3) were good α-glucosidase inhibitors (IC(50) = 6.0-42.6 μM). In kinetic analysis, the most potent neuraminidase inhibitors (5-10) were noncompetitive. HPLC analysis indicated that most pterocarpan synthesis began from the R3 stage, and a rapid change of pterocarpan concentrations was observed between the R4 and R7 stages.     

Comparative analysis of physicochemicals and antioxidative properties in new red rice (Oryza sativa L. cv. Gunganghongmi)

The main objectives of this study were to investigate physicochemicals and antioxidant activities of new red rice (Oryza sativa cv. Gunganghongmi (GH)) by comparing normal brown (Nampyeongbyeo, NB) and reported red rice (Jukjinjubyeo, JB) in Korea. The nutritional constituents, including protein, oil, sugar, fatty acid, GABA, and γ-oryzanol were not significantly different between normal brown and colored rice. However, the ethanol extract of GH showed the highest phenolic content (24.7 ± 1.3 mg g^?1). The ethanol extracts of GH showed higher scavenging activities against DPPH (0.2 mg mL^?1 = 62.1 ± 2.5%) and ABTS (0.2 mg mL^?1 = 63.2 ± 3.5%) radicals. Moreover, GH more inhibited LPS-induced nitric oxide (NO) production (13.2 ± 1.4 μM) than JB (18.3 ± 2.3 μM) and NB (22.1 ± 1.4 μM) at the same concentration (0.2 mg mL^?1) without cytotoxicity. These results suggest that new red rice (GH) would be considered to be new functional rice due to its anti-oxidative effect and high nutrition.     

Culture results from milk samples submitted to veterinary diagnostic laboratories from August 2003 to December 2006 in New Zealand

Abstract

AIMS: To determine the pattern of isolation of major mastitis-causing organisms isolated from milk samples submitted to five veterinary diagnostic laboratories in New Zealand.

METHODS: The culture results of 25,288 milk samples that were collected from dairy cows throughout New Zealand from August 2003 to December 2006 and submitted to a group of veterinary diagnostic laboratories were assembled, reviewed and summarised. Logistic regression was used to analyse the effect of year, region (i.e. North vs South Island), and season on the probability of isolating the two most common organisms.

RESULTS: The most commonly isolated mastitis causing organisms from all samples were: Streptococcus uberis (23.6%), Staphylococcus aureus (23.5%), coagulase-negative staphylococci (CNS; 7.2%), Strep. dysgalactiae (6.2%), Bacillus spp. (4.0%), and coliforms (3.7%). The percentage of samples with isolates of Strep. uberis or Staph. aureus was affected by island, year and season (p<0.001). For most of the year, except in late winter and early spring when Strep. uberis was much more common, the percentage of isolates of Strep. uberis and Staph. aureus were not apparently different despite the former being an environmental pathogen and the other a contagious one.

CONCLUSION: The pattern of isolation of major mastitis-causing organisms, as determined from culture of milk samples submitted to diagnostic laboratories in New Zealand, has changed significantly over the last 40 years, with a substantial increase in the percentage of isolates that are Strep. uberis and a decrease in isolates of Strep. agalactiae. There is a clear seasonal pattern to the isolation of both Strep. uberis and Staph. aureus, particularly the former.

CLINICAL RELEVANCE: Knowledge of the aetiological agents causing bovine mastitis on a farm is of value in determining the choice of treatment. This dataset shows that, although there is seasonal pattern to the isolation of mastitis-causing organisms in New Zealand, both Strep. uberis and Staph. aureus are isolated throughout the year, so bacteriology is of value in determining aetiology even in late winter/early spring.     

Harbour seal (Phoca vitulina) abundance within the Firth of Tay and Eden Estuary,Scotland: recent trends and extrapolation to extinction

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Decomposition of cattle dung on grazed signalgrass (Brachiaria decumbens Stapf) pastures in monoculture or intercropped with tree legumes

Livestock excreta is one of the major nutrient sources in natural grasslands. Understanding how livestock diet and season affects the decomposition dynamics is critical to nutrient cycling models. We hypothesised that livestock diet and season of the year affect dung decomposition. This study evaluated the decomposition and release of N, P, K, Ca, Mg, and Na from faeces of cattle collected in dry and wet seasons. Treatments were signalgrass (Brachiaria decumbens Stapf) in monoculture or mixed with sabiá (Mimosa caesalpiniifolia Benth.) or gliricidia (Gliricidia sepium (Jacq.) Kunth ex Walp.). Excreta samples were incubated in nylon bags for 0, 2, 4, 8, 16, 32, 64, 128 and 256 d. The single negative exponential mathematical model was adequate (P < 0.0001) to explain the decomposition. The relative rate of dung decomposition (k) was greater for samples that originated from cattle grazed on signalgrass (k = 0.00284 g g^?1 d^?1), followed by sabiá (k = 0.00233 g g^?1 d^?1), and gliricidia (k = 0.00200 g g^?1 d^?1) pastures. The rate of nutrient release showed a time effect for all variables and interaction between time and season for Ca (P = 0.0042) and Mg (P = 0.0013). Faeces collected from cattle grazing/browsing intercropped pastures tended to have lower decomposition rates.     

Optimum Dietary Protein Level and Protein‐to‐energy Ratio for Growth of Juvenile Parrot Fish,Oplegnathus fasciatus

An 8‐wk feeding trial was conducted to estimate the optimum dietary protein level and protein‐to‐energy (P/E) ratio in juvenile parrot fish, Oplegnathus fasciatus. Eight experimental diets were formulated with two energy levels and four protein levels for each energy level. Diets containing crude protein (CP) at 35, 40, 45, and 50% had either 12.5 or 14.6 kJ/g of energy. Fish averaging 7.1 ± 0.06 g (mean ± SD) were fed one of the experimental diets for 8 wk. At the end of the feeding trial, weight gain (WG) of fish fed 45 and 50% CP in the 12.5 kJ/g diet was significantly higher than fish fed the 35% CP diet (P < 0.05). WG of the fish fed 45 and 50% CP in the 14.6 kJ/g diet was significantly higher than fish fed the 35 and 40% CP diets (P < 0.05). Fish fed the 14.6 kJ/g diet had a higher WG compared with fish fed the 12.5 kJ/g diet at all CP levels. Feed efficiency (FE) and specific growth rate (SGR) showed a similar trend to the WG. WG, FE, and SGR improved with increasing dietary protein levels up to 45% and remained constant at 50% CP for both energy levels. However, protein efficiency ratio was negatively related to dietary protein levels. The results suggested that the optimum level of protein and the optimum P/E ratio for juvenile parrot fish should be 45% and 31.1 mg protein/kJ, respectively, in a diet containing 14.6 kJ/g energy.