No abstract

Resistance to diflubenzuron in the Australian sheep blowfly, Lucilia cuprina , has rendered this insecticide incapable of preventing flystrike in sheep from a few districts in eastern Australia. Wool producers affected by this situation must find suitable alternatives to protect their flocks. Results of laboratory bioassays against one population demonstrated that, despite extremely high diflubenzuron resistance (Resistance Factor > 791), it had only very low (2x) tolerance of cyromazine and dicyclanil. It is unlikely that this level of tolerance would have any practical impact on field control with either insecticide. Consequently, wool producers in districts where diflubenzuron-resistant flies are common can rotate insecticide treatment to either of these compounds to prevent flystrike in their flocks. However, unlike the highly diflubenzuron-resistant field strain, a laboratory strain selected for resistance to diflubenzuron (Resistance Factor = 617) was 10 times more resistant to dicyclanil than a susceptible strain but, like the field strain, was only two times more tolerant of cyromazine. Conversely, a field-derived strain selected in the laboratory for cyromazine resistance was 20 times more resistant to dicyclanil and 362 times more resistant to diflubenzuron than the reference susceptible strain.

This study was carried out with fresh Australian lager beer which was sampled directly off the production line, the same samples aged for 12 weeks at 30 °C, and the vintage beer which was kept at 20 °C for 5 years. Characteristic Australian lager flavour was maintained in the fresh and vintage beers but was lost in the aged beer. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) and free thiol group labelling analyses of beer proteins found that this flavour stability correlated with the presence of an unknown 10 kilodaltons (kDa) protein with a higher level of free thiols. The protein was purified by size-exclusion chromatography, then peptide sequencing and database matching identified it as the barley lipid transfer protein (LTP1). Further characterisation using diphenylpicrylhydrazyl (DPPH) free radical scavenging and a Saccharomyces cerevisiae-based antioxidant screening assay demonstrated that the LTP1 protein was active in DPPH reduction and antioxidant activity. The absence of free thiol in the aged beer indicates that the thiol functional groups within the LTP1 protein were saturated and suggests that it is important in the flavour stability of beer by maintaining reduction capacity during the ageing process.

BackgroundBovine theileriosis, caused by the haemoprotozoan Theileria orientalis, is an emerging disease in East Asia and Australasia. Previous studies have demonstrated transplacental transmission of various Theileria spp. but molecular confirmation of transplacental transmission of T. orientalis has never been confirmed in the field. In this study, cow-calf (< 48 h old) pairs were sampled across 3 herds; opportunistic samples from aborted foetuses or stillborn calves were also examined. Molecular (multiplex qPCR) and serological (ELISA) methods were used to determine infection prevalence and the presence of anti-Theileria antibodies in each herd. In addition, pregnant heifers and foetal calves were sampled at abattoir and tested for the presence of T. orientalis by qPCR.ResultsThe qPCR results indicated that, even though there was a high prevalence of T. orientalis infection in cows, the rate of transplacental transmission to their calves was low, with only one newborn calf from one herd and one foetus from the abattoir testing positive for T. orientalis DNA. Five aborted foetuses and stillborn calves, 3 of which were derived from a herd experiencing a high number of clinical theileriosis cases at the time of sampling, all tested negative for T. orientalis by qPCR. This suggests that in utero infection of calves with T. orientalis may not be a major driver of abortions during theileriosis outbreaks. Temporal monitoring of 20 calves born to T. orientalis-positive mothers indicated that T. orientalis was detectable in most calves between 10 and 27 days post-partum, consistent with prior field studies on adult cattle introduced to Theileria-affected herds. There was a positive correlation between the ELISA ratio of newborn calves and their mothers within 48 h of calving; however, maternal antibodies were only detectable in some calves and only for 4–4.5 weeks post-partum. All calves displayed high parasite loads peaking at 4–8 weeks post-partum, with only some calves subsequently mounting a detectable adaptive antibody response.ConclusionsThese findings indicate transplacental transmission of T. orientalis appears to play only a minor role in persistence of T. orientalis infection in the field; however calves are highly susceptible to developing high level T. orientalis infections at 4–8 weeks of age regardless of whether maternal antibodies are present post-partum.Electronic supplementary materialThe online version of this article (doi:10.1186/s13071-017-2166-9) contains supplementary material, which is available to authorized users.

In vitro monooxygenase activity (aldrin epoxidation) in 19 field-collected strains of the Australian sheep blowfly Lucilia cuprina (Wiedemann) varied over a 46-fold range. Activities were significantly correlated with toxicological responses to diflubenzuron and diazinon. Relationships between activity and toxicological response were stronger at the LC95 than at the LC50. Toxicological responses to diflubenzuron and diazinon were significantly related, particularly at the LC95. The slope of diflubenzuron concentration-response lines decreased as enzyme activity increased, suggesting that the proportion of the larval population that can tolerate high rates of diflubenzuron increases with increasing mean enzyme levels. Tolerance levels to diflubenzuron among the field strains (relative to a reference susceptible strain) were up to 10-fold at LC50 and 56-fold at LC95. This tolerance appears to be provided, at least in part, by enhanced larval monooxygenase levels.

Late in 2017, field samples of the Australian sheep blowfly, Lucilia cuprina , were submitted by sheep producers from three states of Australia (South Australia, Victoria and New South Wales). Some were collected by submitters concerned about shortened periods of flystrike protection from dicyclanil based products. Neonate larval offspring from the NSW field samples survived and successfully completed their life cycles following exposure to dicyclanil and cyromazine at susceptible discriminating concentrations in vitro . The in vivo study reported here used dicyclanil resistant neonate larvae to assess the flystrike protection provided by a cyromazine jetting fluid and a number of dicyclanil based spray-on products, when applied to sheep six weeks after shearing. The two dicyclanil resistant blowfly strains used in this study showed in vitro resistance ratios, at the LC50, of approximately 13- and 25-fold relative to a dicyclanil and cyromazine susceptible strain. Compared to the levels of resistance that L. cuprina has developed to other insecticides these are relatively low, however, three dicyclanil based spray–on products (active ingredient 12.5 g/L, 50 g/L and 65 g/L) had protection periods reduced by 73%, 78% and 69% respectively when compared to the maximum protection periods claimed by the manufacturer. A 50% and a 33% reduction in protection period was also observed to a cyromazine and an ivermectin based jetting fluid respectively. In contrast, protection periods were attained or exceeded regardless of the treatment used against field derived dicyclanil susceptible neonate larvae. For the first time we confirm that dicyclanil resistance enables the completion of the L. cuprina life cycle following flystrike initiation on dicyclanil or cyromazine treated sheep when insecticide levels are considered high and protective. This study also provides in vivo information on the effect of dicyclanil resistance on the protection provided by a product with an active ingredient belonging to an unrelated insecticide group. Dicyclanil resistance is of major concern to the Australian sheep industry.

Cross-resistance spectra were determined in strains of the Australian sheep blowfly, Lucilia cuprina (Wiedemann), which had been pressured for several years in the laboratory with diflubenzuron, butacarb or deltamethrin. Each strain was highly resistant to its selecting chemical (resistance factors > 1000-fold), however, cross-resistance levels were variable and often low. In particular, strains selected with diflubenzuron and butacarb showed very little resistance to deltamethrin (resistance factors < 7-fold). Each strain showed resistance levels to diazinon only slightly higher than the highest levels currently detected in field strain larvae. Piperonyl butoxide and triphenyl phosphate significantly synergized each pressured strain with its selecting chemical, suggesting the involvement of both monooxygenases and esterases in the observed resistances. Synergism ratios in each case were greater with piperonyl butoxide. The lack of any alteration in in vitro acetylcholinesterase sensitivity to butacarb inhibition in the butacarb-selected strain, and only low level resistance to DDT in the deltamethrin-selected strain, provided no evidence for target-site insensitivities in these strains. The low-moderate levels of cross-resistance therefore imply the existence of qualitative differences in the detoxification systems in each strain.

Chlamydia pecorum is an obligate intracellular pathogen with a wide host range including livestock such as sheep, cattle, goats, and pigs as well as wildlife species such as koalas. Chlamydial polyarthritis is an economically important disease resulting in swollen joints, lameness, stiffness, and weight loss in young sheep. In the present study, tissues from sheep experimentally or naturally infected with Chlamydia pecorum were assessed by histopathology and immunohistochemistry. Carpal, hock, and stifle joints as well as spleen, liver, kidney, lymph nodes, lung, and brain of 35 sheep from different inoculation groups were available. Two different C. pecorum strains (IPA and E58), different routes of administration (intraarticular or intravenous), UVA-irradiated IPA strain, and corresponding noninfected control groups were investigated. Similar investigations on tissues from 5 naturally infected sheep were performed. The most obvious inflammatory lesions were observed in synovial tissues and, notably, in the renal pelvis from the experimentally infected group and naturally infected animals. This resulted in chronic or chronic-active arthritis and pyelitis. Intralesional chlamydial inclusions could be demonstrated by immunohistochemistry in both tissues. Immunohistochemical evaluation of the presence and distribution of macrophages, T and B cells in synovial tissues revealed macrophages as the most prevalent inflammatory cell population. Previous observations indicated that C. pecorum isolates can infect circulating monocytes. Together with the finding of the histological lesions in synovial tissues and internal organs alongside the presence of C. pecorum DNA, these observations suggest chlamydial arthritis in lambs is the result of hematogeneous spread of C. pecorum.