The calicheamicin gene cluster and its iterative type I enediyne PKS

The enediynes exemplify nature's ingenuity. We have cloned and characterized the biosynthetic locus coding for perhaps the most notorious member of the nonchromoprotein enediyne family, calicheamicin. This gene cluster contains an unusual polyketide synthase (PKS) that is demonstrated to be essential for enediyne biosynthesis. Comparison of the calicheamicin locus with the locus encoding the chromoprotein enediyne C-1027 reveals that the enediyne PKS is highly conserved among these distinct enediyne families. Contrary to previous hypotheses, this suggests that the chromoprotein and nonchromoprotein enediynes are generated by similar biosynthetic pathways.     

Lovastatin as a supplement to mitigate rumen methanogenesis:an overview

Methane from enteric fermentation is the gas with the greatest environmental impact emitted by ruminants.Lovastatin(Lv)addition to feedstocks could be a strategy to mitigate rumen methane emissions via decreasing the population of methanogenic archaea(MA).Thus,this paper provides the first overview of the effects of Lv supplementation,focusing on the inhibition of methane production,rumen microbiota,and ruminal fermentation.Results indicated that Lv treatment had a strong anti-methanogenic effect on pure strains of MA.However,there are uncertainties from in vitro rumen fermentation trials with complex substrates and rumen inoculum.Solid-state fermentation(SSF)has emerged as a cost-effective option to produce Lv.In this way,SSF of agricultural residues as an Lv-carrier supplement in sheep and goats demonstrated a consistent decrease in ruminal methane emissions.The experimental evidence for in vitro conditions showed that Lv did not affect the volatile fatty acids(VFA).However,in vivo experiments demonstrated that the production of VFA was decreased.Lv did not negatively affect the digestibility of dry matter during in vitro and in vivo methods,and there is even evidence that it can induce an increase in digestibility.Regarding the rumen microbiota,populations of MA were reduced,and no differences were detected in alpha and beta diversity associated with Lv treatment.However,some changes in the relative abundance of the microbiota were induced.Further studies are recommended on:(i)Lv biodegradation products and stability,as well as its adsorption onto the solid matter in the rumen,to gain more insight on the“available”or effective Lv concentration;and(ii)to determine whether the effect of Lv on ruminal fermentation also depends on the feed composition and different ruminants.     

Cloning of a Genomic DNA Encoding Caffeoyl-coenzyme A 3-<Emphasis Type="Italic">O</Emphasis>-methyltransferase of Citrus

In recent studies, we found that Apll (a PthA homologue) bound to three citrus proteins. Amino acid sequence analysis revealed that one of the target proteins was homologous to that of S-adenosyl-l-methionine : trans-caffeoyl-coenzyme A 3-O-methyltransferase (CCoAMT), an enzyme which is specific for the substrate trans-caffeoyl-CoA and catalyzes the synthesis of trans-feruloyl-CoA. From the consensus nucleotide sequences of CCoAMT genes, primers were chosen for PCR amplification of this gene from citrus total DNA. Two selected DNA fragments of 1.0 kb and 2.0 kb were obtained. These fragments were used as the probe to screen a citrus library. One clone, pCCl00, contained a 1.0-kb SalI fragment that hybridized to the probes. The nucleotide sequence of this fragment was determined in both directions. In this fragment, there was an open reading frame of 232 amino acids interrupted by an intron of 106 nt, and the deduced amino acid sequence had 95.9% homology to tobacco CCoAMT. Southern blot analysis of total citrus DNA showed that four EcoRI fragments hybridized to the probes, suggesting the presence of more than one copy of CCoAMT in the citrus DNA. Received 4 November 1999/ Accepted in revised form 26 November 1999     

Characterization of whitebait (Galaxias maculatus) respiratory rates to optimize intensive culture carrying capacities

Galaxias maculatus is an osmeriform native fish of the Southern Hemisphere, in which the crystalline larvae is considered as a luxury delicacy, for this reason, it has been commercially exploited in Chile, Argentina and New Zealand. However, the fisheries have been rapidly decreasing due to the overexploitation and the predation of introduced species. Because of these events, there is a need to determine a carrying capacity for an intensive fish culture. In order to optimize stocking densities for fish culture, this paper proposes objectives to determine oxygen consumption (OC) rates, dissolved oxygen concentrations that produce signs of hypoxia and the average time elapsed between food intake and peak OC in G. maculatus. In the oxygen experiments under routine metabolism conditions, we found that G. maculatus adults and whitebait showed signs of asphyxia at dissolved oxygen concentrations between 1.3 and 2.2 mg L^?1 and that adults tolerated dissolved oxygen levels as low as 1.3 mg L^?1. The results showed that G. maculatus individuals with an average weight of 0.04 g consumed 0.048 mg O₂ h^?1, whereas individuals with an average weight of 1.4 g consumed 0.345 mg O₂ h^?1. Galaxias maculatus increased the OC rate by 31%, from 0.39 to 0.51 mg O₂ h^?1 g^?1, occurring 14 min after food intake. The carrying capacities for industrial cultures of G. maculatus, were estimated using an allometric equation (OC=0.2363 ×W ^0.612 ), a water flow rate of 1 m³ h^?1 and an input oxygen concentration of 10 mg L^?1 at 12 °C. The density culture of whitebait (4 g) can be allowed to reach 8–11 kg m^?3; therefore, these stocking densities reduce the risk of hypoxia and mortality, ensuring the appropriate growth and feed conversion rates.     

Dietary Lipid Requirements for Optimal Egg Quality of Redclaw Crayfish,Cherax quadricarinatus

The effect of three crude dietary lipid levels (4, 8, and 12%) on egg quality of the female redclaw crayfish, Cherax quadricarinatus, was evaluated. Three replicate groups of 40 crayfish (mean initial weight ± SD, 23.0 ± 3.0 g) per diet treatment were stocked in 15,000 ‐L tanks, at 28 C. After 75 d of culture, there were significant differences (P < 0.001) in egg area (3.52–3.90 mm²), volume (34.0–39.3 mm³), weight (4.81–5.36 mg), and diameter (2.15–2.27 mm) in response to dietary lipids in the diet. Using the quadratic equation, the maximum responses of these parameters corresponded to lipid levels from 8.60–8.84%. There were no significant differences (P > 0.05) in survival (82.1–86.9%), final weight (41.0–43.7 g), number of spawning females (18.12–27.38%), and fecundity (9.07–10.31 eggs/g female). Additionally, there were no significant differences (P > 0.05) in egg mean protein (2306.87 ± 440.08 μg/egg), lipid (441.95 ± 83.23 μg/egg) and carbohydrate (74.81 ± 10.12 μg/egg) contents, and energy (13.49 ± 2.25 kcal/egg). The optimum overall response was obtained for dietary lipid content of 8.7%, which we recommend to optimize egg quality of this freshwater crayfish.