ALBERT

Notes: An experiment was carried out with Cichlasoma urophthalmus (Günther) juveniles to determine the phosphorus requirement and its interaction with dietary calcium. Twelve isoenergetic and isoproteic diets were prepared using a basal artificial diet containing vitamin-free casein, dextrin, starch, corn oil, fish oil, vitamin mixture and a mineral mixture free of calcium and phosphorus. Calcium and phosphorus levels were determined in the casein. To the basal diets were added different concentrations of phosphorus as potassium monophosphate (0.5, 1.0, 1.5 and 2.5 g kg–1) and calcium as calcium carbonate (0.5, 0.75, 1.0, 1.5, 2.0, 2.5, 3.0 and 4.0 g kg–1). These concentrations resulted in varying Ca–P ratios (1:1, 1.33:1, 1.5:1, 1.6:1 and 2.0:1). Calcium and phosphorus concentrations in the water were 84 mg kg–1 and 0.003 mg kg–1, respectively. The diet with 0.5 g kg–1 phosphorus resulted in deficiency signs such as reduced growth, high conversion ratio, high fat content and low bone mineralization. Increased levels of dietary calcium and phosphorus both gave improved growth and mineralization. Mineralization continued to increase with dietary phosphorus levels above that required for maximum growth. The optimum level of phosphorus in the diet was 1.5 g kg–1, the optimum calcium level was 1.8 g kg–1 and the optimum Ca–P ratio was 1.3. Carcass lipid levels were inversely related to dietary phosphorus.

Notes: The effect of the proportion of Calluna vulgaris cover on diet composition, intake and performance of sheep grazing hill vegetation communities in northern Spain is examined. A total of 591 non-lactating Gallega ewes grazed for five consecutive grazing seasons (June to September) on replicated plots of hill pastures (1700 m.a.s.l.) composed principally of Festuca, Agrostis, Nardus and Calluna spp. but with different proportions of Calluna vulgaris cover, either 0·3 (C0·3) or 0·7 (C0·7) of the total area. In 1 year, twenty-eight ewes suckling single lambs also grazed the plots. The mean stocking density over the 5 years was 8·7 ewes ha–1. On treatment C0·3, daily liveweight gains (33 g d–1) of non-lactating ewes were significantly (P 〈 0·001) greater than on treatment C0·7 (12 g d–1). Likewise in lactating ewes the difference in mean daily liveweight change was 40 g d–1 (–5 vs. –45 g d–1 for C0·3 and C0·7 treatments respectively; P 〈 0·001). Liveweight gains of lambs were only 80–100 g d–1 from June to August and lambs only maintained live weight during August and September. The effect of lactational status on liveweight changes was not significant. Liveweight gains of non-lactating ewes increased significantly (P 〈 0·001) from the first to the last year of the experiment on both treatments.The composition of the diet was significantly affected by treatment (P 〈 0·001), with a higher proportion of grass species on the C0·3 treatment and a higher digestibility of the diet in the first half of the grazing season (P 〈 0·001). The proportion of C. vulgaris in the diet was significantly (P 〈 0·001) higher on the C0·7 treatment and increased significantly (P 〈 0·001) from July to September on both treatments. There were no significant differences in the composition of the diet selected by lactating and non-lactating ewes.The results demonstrate that on hill vegetation communities, in which the grass components (Festuca rubra, Agrostis capillaris, and Nardus stricta) cover at least 0·3 of the area and on which the preferred grass component (Festuca and Agrostis spp.) is maintained at a sward height of at least 2·5 cm, non-lactating ewes can increase their live weight and body condition, but this increase is influenced by the proportion and quantity of species of grass in the diet, which is affected in turn by the species of grass available and their nutritive quality. However, ewes suckling lambs were not able to maintain their live weight and body condition except when Calluna cover was 0·3 and grass height was more than 3·5 cm.It is concluded that these indigenous vegetation communities can be used in sheep production systems to complement the use of improved pastures at other times of year. In particular, they can be utilized during the non-lactating period (summer) to increase body condition before the beginning of the mating period in autumn.

Notes: Nitrogen metabolism in Aspergillus nidulans is regulated by AREA, a member of the GATA family of transcription factors. One mechanism that modulates AREA activity involves the rapid degradation of the areA transcript when sufficient NH4+ or Gln are available. This signalling mechanism has been shown to require a region of 218 nucleotides within the 3′ untranslated region of areA mRNA. We demonstrate that this region functions independently in a heterologous transcript and acts to accelerate degradation of the poly(A) tail, which in turn leads to rapid transcript degradation in response to the addition of NH4+ or Gln to the growth medium. areA transcript degradation is inhibited by cycloheximide, but this is not a general consequence of translational inhibition. We believe that this is the first reported example in which specific physiological signals, acting through a defined sequence within a transcript, have been shown to promote accelerated poly(A) degradation, which in turn triggers transcript degradation.

Notes: Saccharomyces cerevisiae PAU genes constitute the largest multigene family in yeast, with 23 members located mainly in subtelomeric regions. The role and regulation of these genes were previously unknown. We detected PAU gene expression during alcoholic fermentation. An analysis of PAU gene regulation using PAU–lacZ fusions and Northern analyses revealed that they were regulated by anaerobiosis. PAU genes display, however, different abilities to be induced by anaerobiosis and this appears to be related to their chromosomal localization; two subtelomeric copies are more weakly inducible than an interstitial one. We show that PAU genes are negatively regulated by oxygen and repressed by haem. Examination of PAU gene expression in rox1Δ and tup1Δ strains indicates that PAU repression by oxygen is mediated by an unknown, haem-dependent pathway, which does not involve the Rox1p anaerobic repressor but requires Tup1p. Given the size of the gene family, PAU genes could be expected to be important during yeast life and some of them probably help the yeast to cope with anaerobiosis.

Notes: RmInt1 is a functional group II intron found in Sinorhizobium meliloti where it interrupts a group of IS elements of the IS630-Tc1 family. In contrast to many other group II introns, the intron-encoded protein (IEP) of RmInt1 lacks the characteristic conserved part of the Zn domain associated with the IEP endonuclease activity. Nevertheless, in this study, we show that RmInt1 is capable of inserting into a vector containing the DNA spanning the RmInt1 target site from the genome of S. meliloti. Efficient homing was also observed in the absence of homologous recombination (RecA− strains). In addition, it is shown that RmInt1 is able to move to its target in a heterologous host (S. medicae). Homing of RmInt1 occurs very efficiently upon DNA target uptake (conjugation/electroporation) by the host cell resulting in a proportion of invaded target of 11–30%. Afterwards, the remaining intronless target DNA is protected from intron invasion.

Notes: We have cloned and sequenced the Lactobacillus casei ptsH and ptsI genes, which encode enzyme I and HPr, respectively, the general components of the phosphoenolpyruvate–carbohydrate phosphotransferase system (PTS). Northern blot analysis revealed that these two genes are organized in a single-transcriptional unit whose expression is partially induced. The PTS plays an important role in sugar transport in L. casei, as was confirmed by constructing enzyme I-deficient L. casei mutants, which were unable to ferment a large number of carbohydrates (fructose, mannose, mannitol, sorbose, sorbitol, amygdaline, arbutine, salicine, cellobiose, lactose, tagatose, trehalose and turanose). Phosphorylation of HPr at Ser-46 is assumed to be important for the regulation of sugar metabolism in Gram-positive bacteria. L. casei ptsH mutants were constructed in which phosphorylation of HPr at Ser-46 was either prevented or diminished (replacement of Ser-46 of HPr with Ala or Thr respectively). In a third mutant, Ile-47 of HPr was replaced with a threonine, which was assumed to reduce the affinity of P–Ser–HPr for its target protein CcpA. The ptsH mutants exhibited a less pronounced lag phase during diauxic growth in a mixture of glucose and lactose, two PTS sugars, and diauxie was abolished when cells were cultured in a mixture of glucose and the non-PTS sugars ribose or maltose. The ptsH mutants synthesizing Ser-46–Ala or Ile-47–Thr mutant HPr were partly or completely relieved from carbon catabolite repression (CCR), suggesting that the P–Ser–HPr/CcpA-mediated mechanism of CCR is common to most low G+C Gram-positive bacteria. In addition, in the three constructed ptsH mutants, glucose had lost its inhibitory effect on maltose transport, providing for the first time in vivo evidence that P–Ser–HPr participates also in inducer exclusion.