Decoration of Pasteurella multocida lipopolysaccharide with phosphocholine is important for virulence

Phosphocholine (PCho) is an important substituent of surface structures expressed by a number of bacterial pathogens. Its role in virulence has been investigated in several species, in which it has been shown to play a role in bacterial adhesion to mucosal surfaces, in resistance to antimicrobial peptides, or in sensitivity to complement-mediated killing. The lipopolysaccharide (LPS) structure of Pasteurella multocida strain Pm70, whose genome sequence is known, has recently been determined and does not contain PCho. However, LPS structures from the closely related, virulent P. multocida strains VP161 and X-73 were shown to contain PCho on their terminal galactose sugar residues. To determine if PCho was involved in the virulence of P. multocida, we used subtractive hybridization of the VP161 genome against the Pm70 genome to identify a four-gene locus (designated pcgDABC) which we show is required for the addition of the PCho residues to LPS. The proteins predicted to be encoded by pcgABC showed identity to proteins involved in choline uptake, phosphorylation, and nucleotide sugar activation of PCho. We constructed a P. multocida VP161 pcgC mutant and demonstrated that this strain produces LPS that lacks PCho on the terminal galactose residues. This pcgC mutant displayed reduced in vivo growth in a chicken infection model and was more sensitive to the chicken antimicrobial peptide fowlicidin-1 than the wild-type P. multocida strain

Comparative productivity of irrigated short-term ryegrass (Lolium multiflorum) pasture receiving nitrogen, grown alone or in a mixture with white (Trifolium repens) and Persian (T. resupinatum) clovers

Dairy farms in subtropical Australia use irrigated, annually sown short-term ryegrass (Lolium multiflorum) or mixtures of short-term ryegrass and white (Trifolium repens) and Persian (shaftal) (T. resupinatum) clover during the winter-spring period in all-year-round milk production systems. A series of small plot cutting experiments was conducted in 3 dairying regions (tropical upland, north Queensland, and subtropical southeast Queensland and northern New South Wales) to determine the most effective rate and frequency of application of nitrogen (N) fertiliser. The experiments were not grazed, nor was harvested material returned to the plots, after sampling. Rates up to 100 kg N/ha.month (as urea or calcium ammonium nitrate) and up to 200 kg N/ha every 2 months (as urea) were applied to pure stands of ryegrass in 1991. In 1993 and 1994, urea, at rates up to 150 kg N/ha.month and to 200 kg N/ha every 2 months, was applied to pure stands of ryegrass; urea, at rates up to 50 kg N/ha.month, was also applied to ryegrass-clover mixtures. The results indicate that applications of 50-85 kg N/ha.month can be recommended for short-term ryegrass pastures throughout the subtropics and tropical uplands of eastern Australia, irrespective of soil type. At this rate, dry matter yields will reach about 90% of their potential, forage nitrogen concentration will be increased, there is minimal risk to stock from nitrate poisoning and there will be no substantial increase in soil N. The rate of N for ryegrass-clover pastures is slightly higher than for pure ryegrass but, at these rates, the clover component will be suppressed. However, increased ryegrass yields and higher forage nitrogen concentrations will compensate for the reduced clover component. At application rates up to 100 kg N/ha.month, build-up of NO3--N and NH4+-N in soil was generally restricted to the surface layers (0-20 cm) of the soil, but there was a substantial increase throughout the soil profile at 150 kg N/ha.month. The build-up of NO3--N and NH4+-N was greater and was found at lower rates on the lighter soil compared with heavy clays. Generally, most of the soil N was in the NO3--N form and most was in the top 20 cm.