Alternatives to sodium metabisulphite for blackspot prevention in prawns

In Australia, prawns are usually treated with a 1% sodium metabisulphite solution to prevent black spot. Two alternatives, Bacterol and Snow Fresh, were compared to the standard metabisulphite treatment used by industry. Bacterol gave similar protection to sodium metabisulphite, while Snow Fresh showed potential as a substitute. The concentrations most appropriate were determined from residue levels after treatment.

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.

The Biosynthesis of Sesquiterpene Isocyanides and Isothiocyanates in the Marine Sponge Acanthella cavernosa (Dendy); Evidence for Dietary Transfer to the Dorid Nudibranch Phyllidiella pustulosa

The tropical marine sponge Acanthella cavernosa (Dendy) converts potassium [14C] cyanide to axisonitrile-3 (1); this precursor is also used for the synthesis of axisothiocyanate-3 (2) suggesting that isocyanides are precursors to isothiocyanates in A. cavernosa. Likewise, potassium [14C] thiocyanate is used for the synthesis of axisothiocyanate-3; unexpectedly this precursor also labelled axisonitrile-3. These results demonstrate either an interconversion between cyanide and thiocyanate prior to secondary metabolite formation or that the secondary metabolites can themselves be interconverted. Specimens of the dorid nudibranch Phyllidiellu pustulosa, preadapted to a diet of A. cavernosa, fed on 14C-labelled sponges and were subsequently found to contain the radioactive terpenes (1) and (2). Specimens of P. pustulosa, which had not expressed a dietary preference for A. cavernosa in the field, did not generally feed in aquarium tests with 14C-labelled sponges and, therefore, provided non-radioactive extracts. Since control experiments demonstrated the inability of P. pustulosa to synthesise the metabolites de novo, we therefore conclude that P. pustulosa acquires secondary metabolites by dietary transfer from A. cavernosa.

Dietary composition manipulation to enhance the performance of juvenile barramundi (Lates calcarifer Bloch) reared in cool water

Barramundi Lates calcarifer reared in cool water (20-22 degrees C) grow slowly and feed is used poorly compared with fish in warm water (28-32 degrees C). Two comparative slaughter growth assays were carried out with juvenile barramundi to see if increasing the digestible energy (DE) and/or the n-3 highly unsaturated fatty acid (n-3 HUFA) content of the feed would improve growth of fish raised in cool water. Increasing the DE content of the feed from 15 to 17 or 19 MJ kg(-1) while maintaining a constant protein to energy ratio in Experiment 1 brought about significant improvements in feed conversion ratio (FCR) (from 2.01 to 1.19) and daily growth coefficient (DGC; from 0.69 to 1.08%/day) for fish at 20 degrees C. For fish at 29 degrees C, improvements, while significant, were of a lesser magnitude: from 1.32 to 0.97 for FCR and from 3.24 to 3.65%/day for DGC. Increasing the absolute amount of dietary n-3 HUFA, expressed as the sum of eicosapentaenoic and docosahexaenoic fatty acids, from 0.5% to 2.0% in Experiment 2 improved DGC linearly and FCR curvilinearly for fish at 29 degrees C whereas at 20 degrees C, DGC was not affected while FCR improved slightly (from 1.83 to 1.68). Feed conversion ratio was optimized with a dietary n-3 HUFA of about 1.5%. Providing barramundi with a feed that is high in DE (viz 19 MJ kg(-1)) and a digestible protein to DE ratio of 22.5 g MJ(-1) is a practical strategy for improving the productivity of barramundi cultured in cool water whereas increasing dietary n-3 HUFA conferred very little additional benefit.

Persistence of sodium monofluoroacetate (1080) in fox baits and implications for fox management in south-eastern Australia

The persistence of 1080 in two commonly used fox baits, Foxoff ® and chicken wingettes, was assessed under different climatic and rainfall conditions in central-western New South Wales. The rate of 1080 degradation did not change significantly between the Central Tablelands and the relatively hotter and drier environment of the Western Slopes. Loss of 1080 from wingettes was independent of the rainfall and climate conditions, with wingettes remaining lethal to foxes for, on average, 0.9 weeks. Foxoff ® baits remained lethal for longer than wingettes under all tested conditions, although their rate of degradation increased generally with increasing rainfall. As a result, areas baited with Foxoff® will require longer withholding periods for working dogs than those baited with wingettes, especially during dry periods. Wingettes may have advantages for use in sensitive areas where long-term hazards from toxic baits are undesirable. We found significant variations in 1080 concentration in freshly prepared baits that may result in efficacy, non-target and regulatory concerns for baiting campaigns. As a result, the superior quality control and shelf-stability of manufactured Foxoff® may be important criteria for favouring its use over freshly prepared bait types. However, use strategies for any bait type must ensure that foxes consume lethal doses of 1080 to avoid potential problems such as the development of learned aversion to baits or pesticide resistance.

Sulfur-associated polioencephalomalacia in cattle grazing plants in the Family Brassicaceae

Polioencephalomalacia was diagnosed histologically in cattle from two herds on the Darling Downs, Queensland, during July-August 2007. In the first incident, 8 of 20 18-month-old Aberdeen Angus steers died while grazing pastures comprising 60% Sisymbrium irio (London rocket) and 40% Capsella bursapastoris (shepherd's purse). In the second incident, 2 of 150 mixed-breed adult cattle died, and another was successfully treated with thiamine, while grazing a pasture comprising almost 100% Raphanus raphanistrum (wild radish). Affected cattle were either found dead or comatose or were seen apparently blind and head-pressing in some cases. For both incidents, plant and water assays were used to calculate the total dietary sulfur content in dry matter as 0.62% and 1.01% respectively, both exceeding the recommended 0.5% for cattle eating more than 40% forage. Blood and tissue assays for lead were negative in both cases. No access to thiaminase, concentrated sodium ion or extrinsic hydrogen sulfide sources were identified in either incident. Below-median late summer and autumn rainfall followed by above-median unseasonal winter rainfall promoted weed growth at the expense of wholesome pasture species before these incidents.

Identification of a mutation in the para-sodium channel gene of the cattle tick Rhipicephalus (Boophilus) microplus associated with resistance to synthetic pyrethroid acaricides

Resistance against synthetic pyrethroid (SP) products for the control of cattle ticks in Australia was detected in the field in 1984, within a very short time of commercial introduction. We have identified a mutation in the domain II S4-5 linker of the para-sodium channel that is associated with resistance to SPs in the cattle tick Rhipicephalus (Boophilus) microplus from Australia. The cytosine to adenine mutation at position 190 in the R. microplus sequence AF134216, results in an amino acid substitution from leucine in the susceptible strain to isoleucine in the resistant strain. A similar mutation has been shown to confer SP resistance in the whitefly, Bemisia tabaci, but has not been described previously in ticks. A diagnostic quantitative PCR assay has been developed using allele-specific Taqman® minor groove-binding (MGB) probes. Using the assay to screen field and laboratory populations of ticks showed that homozygote allelic frequencies correlated highly with the survival percentage at the discriminating concentration of cypermethrin.

Biodegradation of 1080: Testing soils in south-eastern Australia for sodium fluoroacetate-degrading micro-organisms

Sodium fluoroacetate (1080) is a vertebrate poison commonly used for the control of vertebrate pests in Australia. Long-term environmental persistence of 1080 from baiting operations has likely nontarget species and environmental impacts and is a matter of public concern. Defluorinating micro-organisms have been detected in soils of Western and central Australia, and Queensland, but not in south-eastern Australia. The presence or absence of defluorinating micro-organisms in soils from south-eastern Australia will assist in determining whether long-term environmental persistence of 1080 is or is not occurring. Soils from the Central West Slopes and Plains and Central Tablelands of New South Wales were sampled to investigate the presence and capability of 1080 defluorinating soil micro-organisms. Thirty-one species of micro-organisms were isolated from soils from each site after 10 days incubation in a 20 mM 1080 solution. Of these, 13 isolates showed measurable defluorinating ability when grown in a 1080 and sterile soil suspension. Two species, the bacteria Micromonospora, and the actinomycete Streptosporangium, have not been previously reported for their defluorinating ability. These results indicate that defluorinating micro-organisms are present in soils in south-eastern Australia, which adds weight to other studies that found that 1080 is subject to microbiological degradative processes following removal from the bait substrate. Soil micro-organism defluorination, in combination with physical breakdown and uptake by plants, indicates that fluoroacetate in soils and natural water ways is unlikely to persist. This has implications for the better informed use of 1080 in pest animal management programmes in south-eastern Australia.

Dietary media for mass rearing of rusty grain beetle, Cryptolestes ferrugineus (Stephens) and flat grain beetle, Cryptolestes pusillus (Schonherr) (Coleoptera: Cucujidae)

We developed a suitable diet for mass rearing of Cryptolestes ferrugineus (Stephens) populations under laboratory conditions. Recently, this pest has developed strong level of resistance to phosphine in Australia, and therefore, a significant amount of research has been directed towards its management. In total, nineteen grain-based diets, containing rolled oats, various combinations of cracked grains and flours of wheat, sorghum, maize and barley were tested. Each diet contained a small proportion of wheat germ (4.5% w/w) and torula yeast (0.5% w/w). Experiments were conducted at fixed temperature and relative humidity regimes of 30 ± 2 °C and 70 ± 2%, respectively, and replicated three times. Adults (n = 40) of a laboratory strain of C. ferrugineus were introduced into each diet, removed after 14 days and total numbers of live adult progeny were recorded. The following diets resulted in highest live progeny production: barley flour (95%) (607.67 ± 11.21) = rolled oats (75%) + cracked sorghum (20%) (597.33 ± 33.79) ≥ wheat flour (47.5%) + barley flour (47.5%) (496.67 ± 52.93) > cracked sorghum (95%) (384.00 ± 60.66). The performance of these four diets was then tested with field-collected populations of C. ferrugineus and Cryptolestes pusillus (Schonherr). The diets based on rolled oats + cracked sorghum, wheat flour + barley flour, and barley flour alone consistently produced highest progeny numbers in field-collected populations of both species, with mean progeny numbers ranging from 359.9 to 478.5. The multiplication of C. pusillus was significantly higher than C. ferrugineus on all four diets. Our findings will help in mass rearing of healthy cultures of C. ferrugineus and C. pusillus that will greatly facilitate laboratory and field research and in particular, in developing management tactics for these species.

The insecticidal spider toxin SFI1 is a knottin peptide that blocks the pore of insect voltage-gated sodium channels via a large β-hairpin loop

Spider venoms contain a plethora of insecticidal peptides that act on neuronal ion channels and receptors. Because of their high specificity, potency and stability, these peptides have attracted much attention as potential environmentally friendly insecticides. Although many insecticidal spider venom peptides have been isolated, the molecular target, mode of action and structure of only a small minority have been explored. Sf1a, a 46-residue peptide isolated from the venom of the tube-web spider Segesteria florentina, is insecticidal to a wide range of insects, but nontoxic to vertebrates. In order to investigate its structure and mode of action, we developed an efficient bacterial expression system for the production of Sf1a. We determined a high-resolution solution structure of Sf1a using multidimensional 3D/4D NMR spectroscopy. This revealed that Sf1a is a knottin peptide with an unusually large β-hairpin loop that accounts for a third of the peptide length. This loop is delimited by a fourth disulfide bond that is not commonly found in knottin peptides. We showed, through mutagenesis, that this large loop is functionally critical for insecticidal activity. Sf1a was further shown to be a selective inhibitor of insect voltage-gated sodium channels, consistent with its 'depressant' paralytic phenotype in insects. However, in contrast to the majority of spider-derived sodium channel toxins that function as gating modifiers via interaction with one or more of the voltage-sensor domains, Sf1a appears to act as a pore blocker.

Transfer of Dietary Plant Natural Flavor Compounds From Essential Oils to Maternal Fluids in Pigs

Fetal flavor conditioning during the perinatal stage could be essential at the time of the weaning to reduce the stress and improve the feed intake in pigs. The transfer of flavor compounds from maternal diet to amniotic fluid and milk has been shown in behavioral experiments, but not through analytical procedures such as gas chromatography–mass spectrometry (GC–MS). The aim of the experiment was to trace the principal essential oils compounds supplied in the diet in maternal fluids. Twenty Large White sows around their 104th gestational day were allocated to individual farrowing crates. Two groups of 10 sows were fed either a standard gestation diet or the same diet supplemented with a mix of 8 essential oils at a rate of 1kg/ton during the last 10 days of gestation. At approximately the 113th gestational day, animals were individually treated with 10mg of Lutalyse IM was to induce farrowing. Fresh amniotic fluid was collected during the farrowing in 100-mL glass bottles and immediately stored at −20 °C freezer. During the second lactation day, 10–20 IU of Oxytocin IM was administered to each sow to facilitate collection of milk samples in 20-mL glass bottles. The samples were stored at −20 °C until analyzed by GC–MS. The presence of significant amounts of principal components of all the essential oils except one were found in the milk and amniotic fluid samples of the treated sows relative to the control sows. Our data prove the transfer of selected dietary flavors to maternal fluids and sets the scenario for further trials to manipulate postweaning behavior in piglets.

Non-target species interaction with sodium fluoroacetate (1080) meat bait for controlling feral pigs (Sus scrofa)

Fresh meat baits containing sodium fluoroacetate (1080) are widely used for controlling feral pigs in Queensland, but there is a potential poisoning risk to non-target species. This study investigated the non-target species interactions with meat bait by comparing the time until first approach, investigation, sample and consumption, and whether dying bait green would reduce interactions. A trial assessing species interactions with undyed bait was completed at Culgoa Floodplain National Park, Queensland. Meat baits were monitored for 79 consecutive days with camera traps. Of 40 baits, 100% were approached, 35% investigated (moved) and 25% sampled, and 25% consumed. Monitors approached (P < 0.05) and investigated (P < 0.05) the bait more rapidly than pigs or birds, but the median time until first sampling was not significantly different (P > 0.05), and did not consume any entire bait. A trial was conducted at Whetstone State Forest, southern Queensland, with green-dyed and undyed baits monitored for eight consecutive days with cameras. Of 60 baits, 92% were approached and also investigated by one or more non-target species. Most (85%) were sampled and 57% were consumed, with monitors having slightly more interaction with undyed baits than with green-dyed baits. Mean time until first approach and sample differed significantly between species groups (P = 0.038 and 0.007 respectively) with birds approaching sooner (P < 0.05) and monitors sampling later (P < 0.05) than other (unknown) species (P > 0.05). Undyed bait was sampled earlier (mean 2.19 days) than green-dyed bait (2.7 days) (P = 0.003). Data from the two trials demonstrate that many non-target species regularly visit and sample baits. The use of green-dyed baits may help reduce non-target uptake, but testing is required to determine the effect on attractiveness to feral pigs. Further research is recommended to quantify the benefits of potential strategies to reduce the non-target uptake of meat baits to help improve the availability of bait to feral pigs.