Pesticide residues in food - acute dietary exposure

Consumer risk assessment is a crucial step in the regulatory approval of pesticide use on food crops. Recently, an additional hurdle has been added to the formal consumer risk assessment process with the introduction of short-term intake or exposure assessment and a comparable short-term toxicity reference, the acute reference dose. Exposure to residues during one meal or over one day is important for short-term or acute intake. Exposure in the short term can be substantially higher than average because the consumption of a food on a single occasion can be very large compared with typical long-term or mean consumption and the food may have a much larger residue than average. Furthermore, the residue level in a single unit of a fruit or vegetable may be higher by a factor (defined as the variability factor, which we have shown to be typically ×3 for the 97.5th percentile unit) than the average residue in the lot. Available marketplace data and supervised residue trial data are examined in an investigation of the variability of residues in units of fruit and vegetables. A method is described for estimating the 97.5th percentile value from sets of unit residue data. Variability appears to be generally independent of the pesticide, the crop, crop unit size and the residue level. The deposition of pesticide on the individual unit during application is probably the most significant factor. The diets used in the calculations ideally come from individual and household surveys with enough consumers of each specific food to determine large portion sizes. The diets should distinguish the different forms of a food consumed, eg canned, frozen or fresh, because the residue levels associated with the different forms may be quite different. Dietary intakes may be calculated by a deterministic method or a probabilistic method. In the deterministic method the intake is estimated with the assumptions of large portion consumption of a ‘high residue’ food (high residue in the sense that the pesticide was used at the highest recommended label rate, the crop was harvested at the smallest interval after treatment and the residue in the edible portion was the highest found in any of the supervised trials in line with these use conditions). The deterministic calculation also includes a variability factor for those foods consumed as units (eg apples, carrots) to allow for the elevated residue in some single units which may not be seen in composited samples. In the probabilistic method the distribution of dietary consumption and the distribution of possible residues are combined in repeated probabilistic calculations to yield a distribution of possible residue intakes. Additional information such as percentage commodity treated and combination of residues from multiple commodities may be incorporated into probabilistic calculations. The IUPAC Advisory Committee on Crop Protection Chemistry has made 11 recommendations relating to acute dietary exposure.

Exposure of Tomato Fruit to 1-MCP Improves Quality of Stored Slices

Maintenance of quality, such as firmness, is important during storage of fresh cut produce. This study compared the effects of I-MCP on the quality of tomato slices when intact tomatoes were treated with 1-MCP and then sliced, or tomatoes were sliced and the slices treated with I-MCP. In both instances the MCP treatment was 1 µL Lˉٰ at 20 ºC for 12 h. Tomato cv. 'Revolution' was harvested at the 'pink' stage of maturity, cut into 7-mm slices, and stored as vertical stacks in closed plastic containers at 5ºC for up to 7 days after the 1-MCP treatment. Exposure of intact tomatoes to I-MCP resulted in reduced ethylene production (31%) and firmer (22%) slices than when tomatoes were not I-MCP treated. The application of I-MCP prior to slicing of tomatoes appears a useful strategy to retain quality of stored tomato slices.

Effect of exposure period on degree of dominance of phosphine resistance in adults of Rhyzopertha dominica (Coleoptera: Bostrychidae) and Sitophilus oryzae (Coleoptera: Curculionidae)

Degree of dominance of phosphine resistance was investigated in adults of Rhyzopertha dominica F and Sitophilus oryzae L. Efficacy of the grain fumigant phosphine depends on both concentration and exposure period, which raises the possibility that dominance levels vary with exposure period. New and published data were used to test this possibility in adults of R dominica and S oryzae fumigated for periods of up to 144 h. The concentrations required for control of homozygous resistant and susceptible strains and their F1 hybrids decreased with increasing exposure period. For both species the response lines for the homozygous resistant and susceptible strains and their F1 hybrids were parallel. Therefore, neither dominance level nor resistance factor was affected by exposure period. Resistance was incompletely recessive and the level of dominance, calculated at 50% mortality level, was -0.59 for R dominica and -0.65 for S oryzae. The resistant R dominica strain was 30.9 times more resistant than the susceptible strain, compared with 8.9 times for the resistant S oryzae strain. The results suggest that developing discriminating doses for detecting heterozygote adults of either species will be difficult.

Potential Human Exposure to Australian Bat Lyssavirus, Queensland, 1996-1999

Two human deaths caused by Australian bat lyssavirus (ABL) infection have been reported since 1996. Information was obtained from 205 persons (mostly adults from south Brisbane and the South Coast of Queensland), who reported potential ABL exposure to the Brisbane Southside Public Health Unit from November 1,1996, to January 31, 1999. Volunteer animal handlers accounted for 39% of potential exposures, their family members for 12%, professional animal handlers for 14%, community members who intentionally handled bats for 31%, and community members with contacts initiated by bats for 4%. The prevalence of Lyssavirus detected by fluorescent antibody test in 366 sick, injured, or orphaned bats from the area was 6%. Sequelae of exposure, including the requirement for expensive postexposure prophylaxis, may be reduced by educating bat handlers and the public of the risks involved in handling Australian bats.

Effects of environmental heat on conception rates in lactating dairy cows: Critical periods of exposure

Environmental heat can reduce conception rates (the proportion of services that result in pregnancy) in lactating dairy cows. The study objectives were to identify periods of exposure relative to the service date in which environmental heat is most closely associated with conception rates, and to assess whether the total time cows are exposed to high environmental heat within each 24-h period is more closely associated with conception rates than is the maximum environmental heat for each 24-h period. A retrospective observational study was conducted in 25 predominantly Holstein-Friesian commercial dairy herds located in Australia. Associations between weather and conception rates were assessed using 16,878 services performed over a 21-mo period. Services were classified as successful based on rectal palpation. Two measures of heat load were defined for each 24-h period: the maximum temperature-humidity index (THI) for the period, and the number of hours in the 24-h period when the THI was >72. Conception rates were reduced when cows were exposed to a high heat load from the day of service to 6 d after service, and in wk -1. Heat loads in wk -3 to -5 were also associated with reduced conception rates. Thus, management interventions to ameliorate the effects of heat load on conception rates should be implemented at least 5 wk before anticipated service and should continue until at least 1 wk after service. High autocorrelations existed between successive daily values in both measures, and associations between day of heat load relative to service day and conception rates differed substantially when ridge regression was used to account for this autocorrelation. This indicates that when assessing the effects of heat load on conception rates, the autocorrelation in heat load between days should be accounted for in analyses. The results suggest that either weekly averages or totals summarizing the daily heat load are adequate to describe heat load when assessing effects on conception rates in lactating dairy cows.

Controlled exposure as a management tool for Glasser's disease.

In this study, nasal swabs taken from multiparous sows at weaning time or from sick pigs displaying symptoms of Glasser's disease from farms in Australia [date not given] were cultured and analysed by polymerase chain reaction (PCR). Within each genotype detected on a farm, representative isolates were serotyped by gel diffusion (GD) testing or indirect haemagglutination (IHA) test. Isolates which did not react in any of the tests were regarded as non-typable and were termed serovar NT. Serovars 1, 5, 12, 13 and 14 were classified as highly pathogenic; serovars 2, 4 and 15 being moderately pathogenic; serovar 8 being slightly pathogenic and serovars 3, 6, 7, 9 and 11 being non-pathogenic. Sows were inoculated with the strain of Haemophilus parasuis (serovars 4, 6 and 9 from Farms 1, 2 and 4, respectively) used for controlled challenge 3 and 5 weeks before farrowing. Before farrowing the sows were divided into control and treatment groups. Five to seven days after birth, the piglets of the treatment group were challenged with a strain from the farm which had were used to vaccinate the sows. The effectiveness of the controlled exposure was evaluated by number of piglets displaying clinical signs possibly related to infection, number of antibiotic treatments and pig mortality. Nasal swabs of sick pigs were taken twice a week to find a correlation to infection. A subsample of pigs was weighed after leaving the weaning sheds. The specificity of a realtime PCR amplifying the infB gene was evaluated with 68 H. parasuis isolates and 36 strains of closely related species. 239 samples of DNA from tissues and fluids of 16 experimentally challenged animals were also tested with the realtime PCR, and the results compared with culture and a conventional PCR. The farm experiments showed that none of the controlled challenge pigs showed any signs of illness due to Glasser's disease, although the treatment groups required more antibiotics than the controls. A total of 556 H. parasuis isolates were genotyped, while 150 isolates were serotyped. H. parasuis was detected on 19 of 20 farms, including 2 farms with an extensive history of freedom from Glasser's disease. Isolates belonging to serovars regarded as potentially pathogenic were obtained from healthy pigs at weaning on 8 of the 10 farms with a history of Glasser's disease outbreaks. Sampling 213 sick pigs yielded 115 isolates, 99 of which belonged to serovars that were either potentially pathogenic or of unknown pathogenicity. Only 16 isolates from these sick pigs were of a serovar known to be non-pathogenic. Healthy pigs also had H. parasuis, even on farms free of Glasser's disease. The realtime PCR gave positive results for all 68 H. parasuis isolates and negative results for all 36 non-target bacteria. When used on the clinical material from experimental infections, the realtime PCR produced significantly more positive results than the conventional PCR (165 compared to 86).

Comparative performance of Corymbia hybrids and parental species in subtropical Queensland and implications for breeding and deployment.

Eighty six full-sib Corymbia F1 hybrid families (crosses between C. torelliana and four spotted gum taxa: C. citriodora subsp. variegata, C. citriodora subsp. citriodora, C. henryi and C. maculata), were planted in six trials across six disparate sites in south-eastern Queensland to evaluate their productivity and determine their potential utility for plantation forestry. In each trial, the best-growing 20% of hybrid families grew significantly faster (P=0.05) than open-pollinated seedlots of the parent species Corymbia citriodora subsp. variegata, ranging from 107% to 181% and 127% to 287% of the height and diameter respectively. Relative performance of hybrid families growing on more than one site displayed consistency in ranking for growth across sites and analysis showed low genotype-by-environment interaction. Heritability estimates based on female and male parents across two sites at age six years for height and diameter at breast height, were high (0.62±0.28 to 0.64±0.35 and 0.31±0.21 to 0.69±0.37 respectively), and low to moderate (0.03±0.04 to 0.33±0.22) for stem straightness, branch size, incidence of ramicorns, and frost and disease resistance traits at ages one to three years. The proportion of dominance variance for height and diameter had reduced to zero by age six years. Based on these promising results, further breeding and pilot-scale family forestry and clonal forestry deployment is being undertaken. These results have also provided insights regarding the choice of a future hybrid breeding strategy.

Uneven application can influence the efficacy of s-methoprene against Rhyzopertha dominica (F.) in wheat.

The Juvenile Hormone analogue s-methoprene is used to protect stored grain from pests such as the lesser grain borer, Rhyzopertha dominica (F.). The possibility that uneven application influences s-methoprene efficacy against this species was investigated in the laboratory. Adults of methoprene-susceptible strains were exposed for 14 days to wheat treated at doses of up to 0.6 mg kg-1, or to mixtures of treated and untreated wheat giving equivalent average doses. Adult mortality after exposure to treated wheat was negligible in all cases (3.3%) and there was no significant effect of either average dose or evenness of application. In contrast, the number of adult progeny depended on both the average dose and evenness of application. Average doses of 0.3 and 0.6 mg kg-1 reduced the number of live F1 adults by 99-100% relative to the untreated wheat and no effect of evenness of application was detected. At lower doses, however, efficacy tended to decrease with increasing unevenness of application. When adults from the parental generation were transferred to untreated wheat for another 14 days neither the average dose nor evenness of application in the wheat from which they came had any significant effect on reproduction of these adults. This study demonstrates that uneven application can reduce the efficacy of s-methoprene against R. dominica, but that this is unlikely to influence the performance of s-methoprene against susceptible populations at target doses likely to be used in practice (e.g. 0.6 mg kg-1 in Australia). However, the possibility that uneven application leads to underdosing and selects for resistance should be investigated.

Controlled exposure as a management tool for Glassers disease

Control of bacterial disease of pigs.

Reproduction of phosphine resistant Rhyzopertha dominica (F.) following sublethal exposure to phosphine.

Phosphine fumigation is commonly used to disinfest grain of insect pests. In fumigations which allow insect survival the question of whether sublethal exposure to phosphine affects reproduction is important for predicting population recovery and the spread of resistance. Two laboratory experiments addressed this question using strongly phosphine resistant lesser grain borer, Rhyzopertha dominica (F.). Offspring production was examined in individual females which had been allowed to mate before being fumigated for 48 h at 0.25 mg L -1. Surviving females produced offspring but at a reduced rate during a two-week period post fumigation compared to unfumigated controls. Cumulative fecundity of fumigated females from 4 weeks of oviposition post fumigation was 25% lower than the cumulative fecundity of unfumigated females. Mating potential post fumigation was examined when virgin adults (either or both sexes) were fumigated individually (48 h at 0.25 mg L -1) and the survivors were allowed to mate and reproduce in wheat. All mating combinations produced offspring but production in the first week post fumigation was significantly suppressed compared to the unfumigated controls. Offspring suppression was greatest when both sexes were exposed to phosphine followed by the pairing of fumigated females with unfumigated males and the least suppression was observed when males only were fumigated. Cumulative fecundity from 4 weeks oviposition post fumigation of fumigated females paired with fumigated males was 17% lower than the fecundity of unfumigated adult pairings. Both of these experiments confirmed that sublethal exposure to phosphine can reduce fecundity in R. dominica.

Sublethal Exposure to Phosphine Decreases Offspring Production in Strongly Phosphine Resistant Female Red Flour Beetles, Tribolium castaneum (Herbst)

The red flour beetle is a cosmopolitan pest of stored grain and stored grain products. The pest has developed resistance to phosphine, the primary chemical used for its control. The reproductive output of survivors from a phosphine treatment is an important element of resistance development but experimental data are lacking. We exposed mated resistant female beetles to 0.135 mg/L of phosphine for 48 h at 25°C. Following one week of recovery we provided two non-exposed males to half of the phosphine exposed females and to half of the non-exposed control females. Females that had been exposed produced significantly fewer offspring than non-exposed females. Females that remained isolated produced significantly fewer offspring than both exposed females with access to males and non-exposed controls (P<0.05). Some females were permanently damaged from exposure to phosphine and did not reproduce even when given access to males. We also examined the additional effects of starvation prior to phosphine exposure on offspring production. Non-exposed starved females experienced a small reduction in mean offspring production in the week following starvation, followed by a recovery in the second week. Females that were starved and exposed to phosphine demonstrated a very significant reduction in offspring production in the first week following exposure which remained significantly lower than that of starved non-exposed females (P<0.05). These results demonstrate a clear sublethal effect of phosphine acting on the female reproductive system and in some individuals this can lead to permanent reproductive damage. Pest population rebound after a fumigation may be slower than expected which may reduce the rate of phosphine resistance development. The results presented strongly suggest that phosphine resistance models should include sublethal effects. © 2012 Ridley et al.

Identification of intersectional Corymbia hybrids based on seedling morphology improves with parental divergence

Differences in morphology have provided a basis for detecting natural interspecific hybridisation in forest trees for decades but have come to prominence again more recently as a means for directly measuring gene flow from planted forests. Here we examined the utility of seedling morphology for hybrid discrimination in three hybrid groups relevant to the monitoring of gene flow from plantings of Corymbia (L.D. Pryor & L.A.S. Johnson ex Brooker) taxa in subtropical Australia. Thirty leaf and stem characters were assessed on 907 8-month old seedlings from four parental and six hybrid taxa grown in a common garden. Outbred F1 hybrids between spotted gums (Corymbia citriodora subspecies variegata, C. citriodora subspecies citriodora and Corymbia henryi) tended to more closely resemble their maternal Corymbia torelliana parent and the most discriminating characters were the ratio of blade length to maximum perpendicular width, the presence or absence of a lignotuber, and specific leaf weight. Assignment of individuals into genealogical classes based on a multivariate model limited to a set of the more discriminating and independent characters was highest in the hybrid group, where parental taxa were genetically most divergent. Overall power to resolve among outbred F1 hybrids from both parental taxa was low to moderate, but this may not be a limitation to its likely major application of identifying hybrids in seedlots from native spotted gum stands. Advanced generation hybrids (outbred F2 and outbred backcrosses) were more difficult to resolve reliably due to the higher variances of hybrid taxa and the tendency of backcrosses to resemble their recurrent parents. Visual assessments of seedling morphology may provide a filter allowing screening of the large numbers needed to monitor gene flow, but will need to be combined with other hybrid detection methods to ensure hybrids are detected.