Translation of H. contortus and T. colubriformis from egg to establishment in grazing sheep is unaffected by rainfall timing, rainfall amount and herbage height under conditions of high soil moisture in the Northern Tablelands of NSW.

A field experiment was conducted at Armidale in the Northern Tablelands of NSW, Australia to determine the effects of simulated rainfall amount (0, 12 and 24 mm), rainfall timing (days -1, 0 and 3 relative to plot contamination) and herbage height (4 and 12 cm), on translation of Haemonchus contortus and Trichostrongylus colubriformis from egg to established stages in grazing sheep under conditions of high soil moisture (22-23%). The experiment was conducted in summer when temperature was not anticipated to be a limiting factor for development success. Development success was assessed using tracer sheep and expressed as percentage recovery of parasitic stages relative to egg output on pasture (translation%). For both species, translation (0.11% H. contortus; 0.55% T. colubriformis) was observed in the absence of simulated rainfall and was unaffected by treatment effects of rainfall amount and timing, and herbage height. We suggest that soil moisture (>20%) alone was sufficient to support development and translation (from eggs to parasitic stages in the gut of tracer animals) of these species which contrasts with expectations for development success on dry soils. These findings identify the importance of taking soil moisture into account when predicting the likely effects of rainfall and herbage height on development to L3 and ultimately in predictive epidemiological models of ovine gastrointestinal nematodiasis.

Soil moisture modulates the effects of the timing and amount of rainfall on faecal moisture and development of Haemonchus contortus and Trichostrongylus colubriformis to infective third stage larvae.

Recent experiments on the effects of rainfall and/or soil moisture (SM) on development of sheep gastro-intestinal nematodes to infective L3 stage have used soil of relatively low moisture content in small experimental samples that dry out faster than field soil. To determine whether higher and more sustained SM content modulates the effects of rainfall amount and timing on faecal moisture (FM) and development of H. contortus and T. colubriformis to infective third stage larvae (L3), a climate-controlled chamber experiment was conducted. It was designed to test the effects of rainfall amount (0, 12 and 24 mm), rainfall timing (days -1, 0 and 3 relative to faecal deposition) and soil moisture maintained at 10, 20 and 30% on these variables. Total recovery of L3 14 days after faecal deposition was significantly affected by SM, rainfall timing and their interaction (P<0.01), but not by rainfall amount or species or other two-way interactions. Recovery of L3 was maximal (28%) with a SM treatment of 30% and simulated rainfall on day 3. Faecal moisture was significantly affected by collection day, SM treatment, rainfall amount and rainfall timing with significant interaction between many of these effects (P<0.05). A positive linear association between FM and total L3 recovery was strongest on day 4 after faecal deposition (R(2)=0.64, P<0.001) for H. contortus and day 6 (R(2)=0.78, P<0.001) for T. colubriformis. Overall the results show that SM is able to modulate the effects of rainfall timing and amount with increased SM acting to broaden the window of opportunity for the free-living stages to respond to post deposition rainfall to complete development to L3. If SM is maintained in the range 10-30%, the reported benefits of early rainfall (days -1 and 0) of up to 24 mm appear to be negated with later rainfall (day 3) proving more beneficial. These results require field confirmation.

Soil moisture influences the development of Haemonchus contortus and Trichostrongylus colubriformis to third stage larvae.

Two climate chamber experiments were conducted to determine the effect of varying initial soil moisture (0, 10 and 15%), simulated rainfall amount (0, 12 and 24 mm) and simulated rainfall timing (days -1, 0 and 3 relative to faecal deposition) on development (day 14) of Haemonchus contortus and Trichostrongylus colubriformis to the third stage larvae (L3) and faecal moisture (FM). Increasing initial soil moisture content from 0 to 10 or 15% led to higher recovery of total L3 (P<0.001). Total L3 recovery increased with each level of simulated rainfall (P<0.001) in the ascending order of 0, 12 and 24 mm. There was an interaction between the effects of initial soil moisture and simulated rainfall amount on the recovery of total L3, showing that the benefit of increased simulated rainfall lessened with increasing soil moisture. Simulated rainfall on the day of deposition resulted in higher recovery of L3 (P<0.001) than simulated rainfall on other days. FM on day 3 relative to faecal deposition was best associated with recovery of total H. contortus and T. colubriformis L3 (R(2)=0.32-0.46), reinforcing the importance of sufficient moisture soon after faecal deposition. The effects of initial soil moisture, and the amount and timing of simulated rainfall on development to L3 were largely explained by changes to FM and soil moisture values within 4 days relative to faecal deposition. These results highlight the influence of soil moisture and its interaction with rainfall on development of H. contortus and T. colubriformis to L3. Consequently we recommend that soil moisture be given greater importance and definition in the conduct of ecological studies of parasitic nematodes, in order to improve predictions of development to L3.

Effect of simulated rainfall timing on faecal moisture and development of Haemonchus contortus and Trichostrongylus colubriformis eggs to infective larvae.

Three climate-controlled chamber experiments were conducted to determine the effect of 32 mm simulated rainfall applied prior to (days -4 to -1) or after (days 0-7) faecal deposition and as a single (32 mm) or split (2 × 16 mm) application on faecal moisture (FM) and development of H. contortus and T. colubriformis to third stage infective larvae (L3). The timing of simulated rainfall regulated extra-pellet L3 recovery for H. contortus (P<0.05) but not T. colubriformis. Recovery of L3 was highest (P<0.05) when simulated rainfall was applied on the day of deposition followed by days -1, 1 and 2, which resulted in similar but lower development success rates. Recovery of intra-pellet T. colubriformis L3 was two-fold greater (P=0.008) than for H. contortus and was higher (P=0.007) following simulated rainfall on days 0 and 1 than on other days. There was a positive association between FM and total L3 recovery indicating the importance of FM in the period 48-72 h (H. contortus) and 72-96 h (T. colubriformis) after deposition. Simulated rainfall on the day prior to deposition was as effective in supporting total L3 recovery as application on days 1 or 2 and this effect could be predicted through FM. This highlights the importance of soil in transferring moisture to the faecal pellet. The importance of precedent rainfall and soil moisture in determining the development success of H. contortus and T. colubriformis, in addition to the general effects of the timing of simulated rainfall, need to be accommodated in grazing management programs to combat these species.

Primary structure of mouse, rat, and guinea pig cytochrome c.

For immunochemical and evolutionary reasons we determined the primary structure of cytochrome c from two strains of laboratory mice. Thioacetylthioethane and thioacetylthioglycolic acid were used in addition to conventional reagents for sequence determinations. The sequence was found to be identical with that of the rabbit except for residues 44 and 89 and consistent with the peptide compositional data reported by Hennig (Hennig, B. (1975), Eur. J. Biochem. 55, 167-183). The rat cytochrome c cymotryptic peptides were identical with those of the mouse in amino acid composition and amino-terminal residues. Further, peptide maps of cytochromes c of the guinea pig and two strains of rat indicate that all these animals have the same cytochrome c as the laboratory mouse. It is concluded that rodent cytochromes c are evolutionarily conservative and that there is no evidence for a generation-time effect in cytochrome c evolution.