In vitro efficacy of over-the-counter botanical pediculicides against the head louse Pediculus humanus var capitis based on a stringent standard for mortality assessment

Abstract.  Infestation of the head louse Pediculus humanus var capitis DeGeer (Phthiraptera: Pediculidae) is an important public health problem in Australia, with up to a third of children infested in some primary schools. Insecticide resistance and inadequate attention to the application instructions of topical pediculicides are common reasons for treatment failure. This study evaluated six popular Australian over-the-counter products against head lice, primarily comprised of different botanical extracts, and compared them with permethrin 1% (Quellada^®) and a non-treatment control in order to assess their in vitro efficacy. We also assessed commonly used criteria for evaluating pediculicide efficacy in vitro. All tested products failed to demonstrate high levels of efficacy with the exception of Tea Tree Gel^®, which outperformed 1% permethrin. Permethrin had a high level of efficacy, but using stringent criteria 18% of lice were not dead at 3 h, indicating some resistance to Quellada^®. Commonly used less stringent criteria were shown to overestimate mortality of head lice as a result of the protective phenomenon of stasis or sham death observed in exposed lice that may recover after some time. Using two different levels of stringency resulted in different rankings of efficacy for most products, with the exception of the first ranked product, Tea Tree Gel^®. Rankings of efficacy also varied over time, even within the different assessment criteria. Government regulatory agencies should require standard in vitro tests using stringent mortality criteria, with an observation period of ≥ 6 h, to determine the efficacy of new pediculicides, and only products that cause a minimum mortality rate (e.g. 80%) in head lice collected from the target population should be licensed for sale.     

Bacillus thuringiensis var. tenebrionis shows no toxicity to the predator Chauliognathus lugubris (F.) (Coleoptera: Cantharidae)

Laboratory trials examined the effect of the bioinsecticide Novodor^® (active ingredient 3% Bacillus thuringiensis var. tenebrionis (w/w) (Btt)), on the Australian plague soldier beetle, Chauliognathus lugubris, a predator of the Tasmanian eucalyptus leaf beetle Chrysophtharta bimaculata . Spraying of C. lugubris and the C. bimaculata eggs supplied as food showed that neither Novodor^® FC with active nor inactive Btt caused any decrease in predatory performance, longevity or fecundity of C. lugubris during the period of their population peak (December). Towards the end of their peak population levels, in late January, there was a steady increase in mortality which was not related to either of the Novodor^® FC treatments. On average, male:female pairs of C. lugubris laid one egg batch and ate 30 C. bimaculata eggs a week whether untreated or sprayed with either Novodor^® FC. This Btt-based insecticide appears to be a suitable component of an integrated pest management (IPM) regime that aims to conserve natural predators.     

Food intake and growth in the Moses perch, Lutjanus russelli (Bleeker), with reference to predation on penaeid prawns

Juvenile Moses perch were fed various rations of penaeid prawn and pilchard foods at 26.4 and 28.7° C to elucidate feeding–growth relationships. Maintenance rations amounted to 5.0 and 3.8% of body weight per day (BW day^–1) on prawn and pilchard diets, respectively, at 26.4° C and 4.5% BW day ¹ for prawn at 28.7° C. This apparent reduction in maintenance ration at higher temperature is probably due to reduction in activity levels. Starvation weight loss, an indicator of resting metabolic rate, increased with temperature and exhibited a Q¹⁰ of 2.0 to 2.4.
Fish fed ad libitum once or twice per day at 26.4° C grew at 1.1 % BW day^–1 and ingested about 9% of their BW day^–1. At 28.7° C fish ingested about 8% of their BW day^–1 and grew 1 .0% BW day^–1. These growth rates are comparable to those of similar sized lutjanids in the wild. Since prawns comprise 64% of the diet of Moses perch in northern Australia, this level of consumption would require about 6% BW day^–1 of prawns. The relevance of these findings to the assessment of predator impact on prawns in Australian estuaries is discussed.     

Vector competence of Coquillettidia linealis (Skuse) (Diptera: Culicidae) for Ross River and Barmah Forest viruses

Abstract Coquillettidia linealis is a severe pest on some of the Moreton Bay islands in Queensland, Australia, but little is known of its breeding habitats and biology. Because of its high abundance and its association with Ross River (RR) and Barmah Forest (BF) viruses by field isolation, its vector competence was evaluated in the laboratory by feeding dilutions of both viruses in blood. For RR, Cq. linealis was of comparable efficiency to Ochlerotatus vigilax (Skuse), recognised as being a major vector. Results were as follows for Cq. linealis and Oc. vigilax , respectively: dose to infect 50%, 10^2.2 and <10^1.7 CCID₅₀/mosquito; 88% and 90% disseminated infection at 4 days postinfection; transmission at 4 days with rates of 68−92% and 25−60%. For BF dose to infect 50%, 10^2.7 and 10^2.0; disseminated infection rates on first transmission day (day 6), 40% and 70%; transmission rates of 8−16% and 0−10%. As a capillary-tube method was used rather than suckling mice to demonstrate transmission, transmission rates may be underestimates. This, the first study of the vector competence of Cq. linealis in Australia, demonstrates that this species deserves control on the southern Moreton Bay islands.     

Segregational and structural instability of recombinant plasmid carrying genes for naphthalene degrading pathway

The stability of recombinant plasmid carrying genes for naphthalene mineralization was determined. A strain of Pseudomonas putida capable of mineralizing naphthalene (Nap⁺) via salicylate (Sal⁺) was isolated, and all regulatory and structural genes for the whole pathway were found to be encoded on a 25 kb Eco RI fragment of an approximately 83 kb plasmid present in this strain. The 25 kb Eco RI fragment was cloned into a tetracycline-resistant (Tc^R) cloning vector pLAFR3 and the recombinant plasmid, pRKJ3 (Nap⁺, Sal⁺, Tc^R), thus obtained was transferred into the plasmid-free strain Pseudomonas putida KT2442 in order to test the stability of the plasmid. Plasmid pRKJ3 was found to be segregationally and/or structurally unstable, depending on the growth conditions. Two types of novel derivative strains having the phenotypes Nap⁻, Sal⁺, Tc^R and Nap⁻, Sal⁻, Tc^R with specific deletions of approximately 2 kb and 18 kb, respectively, were obtained.     

Species of Anguilla as indicators of mercury in the coastal rivers and lakes of Victoria, Australia

Mercury concentrations in the axial muscle tissue of most (243) of the 254 Anguilla australis and most (20) of the 27 A. reinhardtii collected from 30 sites in coastal rivers and lakes in Victoria, Australia, during 1975–78 were well below the Australian statutory health limit (0.5 μg g⁻¹ wet weight). For A. australis the mean mercury concentration was 0.17 μg g⁻¹ (±0.16 s.d. , range 0.01–1.60 μg g⁻¹); for A. reinhardtii the values were 0.37 ± 0.23 μg g⁻¹ (range 0.12–1.10 μg g⁻¹). Statistical analyses showed that variation in mercury concentration due to total length accounted for only 13% of the total variation in A. australis and 2% in A. reinhardtii whereas locality accounted for 54 and 68%, respectively. Both species are thus considered suitable as indicators of mercury pollution.     

Calcium/Calmodulin-Dependent Kinase II Phosphorylates Drosophila Visual Arrestin

Abstract: Light activation of rhodopsin in the Drosophila photoreceptor induces a G protein-coupled signaling cascade that results in the influx of Ca²⁺ into the photoreceptor cells. Immediately following light activation, phosphorylation of a photoreceptor-specific protein, phosrestin I, is detected. Strong sequence similarity to mammalian arrestin and electroretinograms of phosrestin mutants suggest that phosrestin I is involved in light inactivation. We are interested in identifying the protein kinase responsible for the phosphorylation of phosrestin I to link the transmembrane signaling to the light-adaptive response. Type II Ca²⁺/calmodulin-dependent kinase is one of the major classes of protein kinases that regulate cellular responses to transmembrane signals. We show here that partially purified phosrestin I kinase activity can be immunodepleted and immunodetected with antibodies to Ca²⁺/calmodulin-dependent kinase II and that the kinase activity exhibits regulatory properties that are unique to Ca²⁺/calmodulin-dependent kinase II such as Ca²⁺ independence after autophosphorylation and inhibition by synthetic peptides containing the Ca²⁺/calmodulin-dependent kinase II autoinhibitory domain. We also show that Ca²⁺/calmodulin-dependent kinase II activity is present in Drosophila eye preparations. These results are consistent with our hypothesis that Ca²⁺/calmodulin-dependent kinase II phosphorylates phosrestin I. We suggest that Ca²⁺/calmodulin-dependent kinase II plays a regulatory role in Drosophila photoreceptor light adaptation.     

Loss of ionic and osmotic regulation in Chara internodal cells in concentrated KCl solutions

Intact internodal cells of Chara are known to maintain their osmotic pressures at constant levels in artificial pond water at room temperature. Cell fragments with osmotic pressures higher and those cell fragments with osmotic pressures lower than the original, both of which are prepared from intact internodal cells using transcellular osmosis and ligation with threads, can also return their osmotic pressures to the original level within a week in artificial pond water. These regulatory phenomena are realized mainly by extrusion of K⁺ and Cl⁻ in the cytoplasm and/or vacuole or by absorption of K⁺ and Cl⁻ from the external solution. According to the electrochemical potential difference calculated for K⁺ between the vacuole and the external solution, the cells should be able to maintain these regulatory functions even in 50–100 m M KCl+ 1 m M CaCl₂ solutions. However, novel phenomena were observed when they were immersed in such concentrated KCl solutions. To maintain electroneutrality, their osmotic pressures increased up to ca l MPa in 2 days due to absorption of K⁺ and Cl⁻ and many gradually died over time. Ionic and osmotic reguratory functions of Chara cells were lost when they were immersed in 50–100 m M K-salt solutions containing 1 m M Ca²⁺.     

Development of Australian commercial producers of invertebrate biological control agents from 1971 to 2014

The Australian industry producing invertebrate biological control agents (BCAs) commercially was described for the first time through interviews with company owners, citrus pest management researchers and citrus growers. This industry started with citrus pest management in 1971. Over 40 years it expanded from one company and one commercial BCA to peak at nine companies operating from 2002 to 2010. A period of consolidation resulted in five companies producing 36 commercial BCAs in 2014. Annual gross sales were <AU$10 million compared to >AU$400 million spent on insecticides. Barriers to industry growth included high costs of developing new commercial BCAs partly due to regulatory costs associated with importation of new BCAs into Australia, and low adoption of commercial BCAs due to limited access to information and training. The industry responded to these barriers by collaborating with stakeholders to develop new commercial BCAs, and developing an accreditation scheme to build technical capacity and encourage adoption.     

Microsatellites reveal a lack of structure in Australian populations of the diamondback moth, Plutella xylostella (L.)

The diamondback moth, Plutella xylostella, is renowned for developing resistance to insecticides and causing significant economic damage to Brassica vegetable crops throughout the world. Yet despite its economic importance, little is known about the population structure and movement patterns of this pest both at local and regional scales. In Australia, the movement patterns and insecticide resistance status of P. xylostella infesting canola, vegetables, forage brassicas and weeds have fundamental implications for the management of this pest. Here we use six polymorphic microsatellite loci to investigate population structure and gene flow in Australian populations of P. xylostella. Samples of P. xylostella from New Zealand, Malaysia, Indonesia and Kenya were also scored at these loci. We found no evidence of population structure within Australia, with most populations having low inbreeding coefficients and in Hardy-Weinberg equilibrium. In addition, a sample from the North Island of New Zealand was indistinguishable from the Australian samples. However, large genetic differences were found between the Australia/New Zealand samples and samples from Kenya, Malaysia and Indonesia. There was no relationship between genetic distance and geographic distance among Australian and New Zealand samples. Two of the loci were found to have null alleles, the frequency of which was increased in the populations outside the Australia/New Zealand region. We discuss these results with reference to insecticide resistance management strategies for P. xylostella in Australia.     

The microbial flora of smoked pork loin and frankfurter sausage stored in different gas atmospheres at 4°C

The development of the microflora of smoked pork loin and frankfurter sausage was followed during storage in vacuum, N₂ and CO₂ atmospheres at 4°C. The total aerobic count on the smoked pork loin reached 10⁷ organisms/g after 37 d in vacuum, 43 d in N₂ and 49 d in CO₂. The corresponding value for the sausage was 77 d in vacuum, while the growth stopped at 6 times 10⁴ organisms/g after 98 d in N₂, and at 4 times 10² organisms/g after 48 d in CO².
The predominant organisms on the fresh products were Bacillus spp., coryneform bacteria, Flavobacterium spp. and Pseudomonas spp.
At the end of the storage time the microflora on both products in the three gas atmospheres, consisted mainly of Lactobacillus spp. and two large groups of organisms that could not be identified as any described genus. Some of the unidentified strains could be classified as a Lactobacillus sp. after subsequent subculturing on laboratory media.
The numbers of Lactobacillus spp. at the end of storage decreased in the order, CO₂ > N₂ > vacuum. Lactobacillus viridescens generally constituted a substantial part of the Lactobacillus flora (5–72%). On the sausages two large uniform groups of unidentifiable homofermentative Lactobacillus spp. were also found.     

Nucleopolyhedrovirus introduction in Australia

Nucleopolyhedrovirus (NPV) has become an integral part of integrated pest management (IPM) in many Australian agricultural and horticultural crops. This is the culmination of years of work conducted by researchers at the Queensland Department of Primary Industries and Fisheries (QDPI&F) and Ag Biotech Australia Pty Ltd. In the early 1970's researchers at QDPI&F identified and isolated a virus in Helicoverpa armigera populations in the field. This NPV was extensively studied and shown to be highly specific to Helicoverpa and Heliothis species. Further work showed that when used appropriately the virus could be used effectively to manage these insects in crops such as sorghum, cotton, chickpea and sweet corn. A similar virus was first commercially produced in the USA in the 1970's. This product, Elcar(R), was introduced into Australia in the late 1970's by Shell Chemicals with limited success. A major factor contributing to the poor adoption of Elcar was the concurrent enormous success of the synthetic pyrethroids. The importance of integrated pest management was probably also not widely accepted at that time. Gradual development of insect resistance to synthetic pyrethroids and other synthetic insecticides in Australia and the increased awareness of the importance of IPM meant that researchers once again turned their attentions to environmentally friendly pest management tools such NPV and beneficial insects. In the 1990's a company called Rhone-Poulenc registered an NPV for use in Australian sorghum, chickpea and cotton. This product, Gemstar(R), was imported from the USA. In 2000 Ag Biotech Australia established an in-vivo production facility in Australia to produce commercial volumes of a product similar to the imported product. This product was branded, ViVUS(R), and was first registered and sold commercially in Australia in 2003. The initial production of ViVUS used a virus identical to the American product but replicating it in an Australian Helicoverpa species, H. armigera. Subsequent research collaboration between QDPI&F and Ag Biotech reinvigorated interest in the local virus strain. This was purified and the production system adapted to produce it on a commercial scale. This new version of ViVUS, which was branded ViVUS Gold(R), was first registered and sold commercially in 2004. Widespread insect resistance to insecticides and a greater understanding of integrated pest management is leading to increased adoption of technologies such NPV in Australian agriculture.     

Variation in nitrate uptake and assimilation between two ecotypes of Lotus japonicus and their recombinant inbred lines

A large genetic variability was observed in the shoot NO₃^– content of recombinant inbred lines (RILs) of Lotus japonicus . To determine the cause of this variability, we have studied some aspects of nitrate uptake and assimilation in the two parental ecotypes (Gifu and Funakura) and four representatives of the RILs population differing both in their shoot biomass and shoot NO₃^– content. Higher shoot NO₃^–content was mainly due to an increase in the uptake of the ion regardless of the plant biomass production. The positive correlation observed between the shoot NO₃^– content and the steady state level of mRNA encoding high affinity NO₃^– transporters suggests that the higher NO₃^– influx is due to enhanced expression of the transporters. In contrast, neither the level of nitrate reductase mRNA, nor the potential enzyme activity in vivo in the different lines was correlated with the shoot NO₃^– content. This indicates that NO₃^– transport in Lotus is one of the main checkpoints controlling shoot NO₃^– accumulation. In addition, this study shows that at least in Lotus , it is possible, through breeding strategies, to lower the NO₃^– content without affecting biomass production.     

Detection of heat-stable enterotoxin genes among Australian Vibrio cholerae O1 strains

Abstract DNA probes derived from the heat-stable enterotoxin gene of Vibrio cholerae non-O1 ( stn ), and the cholera toxin gene (etc), were used to screen 199 strains of V. cholerae O1, which were isolated within Australia from 1977–1986. 13 environmental strains isolated from the riverine environment in Southeast Queensland in 1980 and 1981, hybridized with the stn and ctx DNA probes. The concentrated supernatant of 6 of these strains elicited fluid accumulation in the infant mouse assay both before and after heating at 100 °C for 5 min. Genetic relationships among the 13 stn ⁺ strains were studied by a comparison of the rRNA-RFLPs (ribotyping) and by Southern blot analysis with a stn gene probe. The results indicate that there is a clonal relationship among the Australian stn ⁺ strains and that there is an environmental reservoir of stn genes among Australian V. cholerae O1 isolates.     

Photosynthetic carbon fixation in the corollas of Petunia hybrida

Corollas of Petunia hybrida (cv. Hit Parade Rosa) flowers fixed ¹⁴CO₂ under both light and dark conditions. Rates of light fixation were much higher in mature pink corollas than in young, green corollas [57 and 9 nmol (ngchl)¹ min^-1], paralleling the development of chloroplasts in these tissues. Stomatal conductance in corollas was only 12% of that in green leaves, mainly due to the presence of few, and non-functioning stomata in the corolla. The activity and concentration of ribulose bisphosphate carboxylase (EC 4.1.1.39) in corolla extracts were only about 30% (per unit Chi) of those in extracts from green leaves. These results, together with previous results, might indicate a coordinated reduction in activity of systems participating in photosynthesis in corollas. The fixation products following a 6 s pulse with ¹⁴CO₂, were typical of C, plants in both corollas and green leaves, but a higher level of β-carboxylation products was found in the corollas. The activity of phosphoenol-pyruvate carboxylase (EC 4.1.1.31) (per unit protein) was similar in both tissues. Although the total carbon fixed by the corolla constituted only a small part of the metabolites required for flower development, certain photosynthetic metabolites might have a regulatory role in flower development.     

Determining patterns of use by black bream Acanthopagrus butcheri (Munro, 1949) of re-established habitat in a south-eastern Australian estuary

Acoustic telemetry was used to assess patterns of utilization by black bream Acanthopagrus butcheri of regions with and without re-introduced large woody debris (LWD) in two estuaries of south-eastern Australia, the Mitchell and Tambo Rivers. The fish ( n = 46) were implanted with acoustic transmitters in December 2004 and March 2005 and monitored for c. 12 months. The two principal metrics from the telemetry data, number of visits per day ( N day⁻¹) and residency (amount of time, s) were highly correlated ( r > 0·948), and subsequent analyses were based on N day⁻¹. Rates of N day⁻¹ varied inconsistently among estuarine regions across diel periods and times of year for each river. In the Tambo River during autumn, winter and spring, the N day⁻¹ was greatest in the middle and upper estuarine regions during the day, and often greater in the lower region at dawn and dusk, but varied little among regions in summer. The provision of LWD had little effect on N day⁻¹ in the Tambo River. In the Mitchell River, N day⁻¹ varied little among estuarine regions without LWD, regardless of the time of day, and these patterns were consistent across seasons, but N day⁻¹ was significantly greater to the LWD during the day in winter and spring. Freshwater flows had little effect on monthly patterns of N day⁻¹ among regions in either estuary. The perceived 'benefits' to A. butcheri of re-establishing LWD within estuarine systems of south-eastern Australia depended strongly on the time of year, time of day and river system, but acoustic telemetry was a useful method of evaluating the use by fish of these artificial structures.     

Toxicity of six commercially formulated insecticides and biopesticides to third instar larvae of mushroom sciarid, Lycoriella ingenua Dufour (Diptera: Sciaridae), in New South Wales, Australia

Abstract  Lycoriella ingenua is one of the major pests of cultivated mushrooms, Agaricus bisporus (Lange) Imbach . Insecticide resistance among mushroom sciarid populations has been reported from other countries, and there is a need to determine the toxicity of currently approved and potential pesticides to sustain control of mushroom sciarid populations in Australia. The present study investigated the toxicity of six commercial formulations of insecticides or biopesticides against third instar larvae of L .  ingenua using laboratory bioassays. Insecticide treatments were incorporated into the growing medium for sciarid larvae and the concentration of the pesticide, which killed 90% of the test population (LD₉₀) determined the efficacy of selected insecticides. Triflumuron was the most effective insecticide against L. ingenua with an LD₉₀ of 53.12 mg active ingredient (a.i.)/m² followed by cyromazine (LD₉₀, 179.68 mg a.i./m²) and diazinon (LD₉₀, 261.72 mg a.i./m²). Abamectin and Bacillus thuringiensis ssp. israelensis were ineffective against L. ingenua . Steinernema feltiae , an entomopathogenic nematode, reduced the number of third instar larvae of L. ingenua only when applied at a higher rate (LD₉₀, 732 422 nematodes/m²) than was recommended on the label.     

Compatibility of the insect pathogenic fungus Beauveria bassiana with neem against sweetpotato whitefly, Bemisia tabaci, on eggplant

A study on the compatibility of the entomopathogenic fungus Beauveria bassiana (Balsamo) Vuillemin (Ascomycota: Hypocreales) with neem was conducted against sweetpotato whitefly, Bemisia tabaci (Gennadius) (Hemiptera: Aleyrodidae), on eggplant. Initially, three concentrations of B. bassiana (10⁶, 10⁷, and 10⁸ conidia ml⁻¹) and three concentrations of neem (0.25, 0.5, and 1.0%) were used as individual treatments against B. tabaci . The highest concentration of B. bassiana yielded the highest B. tabaci egg (25.2%) and nymph mortalities (73.0%), but this was not significantly different from the mortalities caused by the 10⁷ conidia ml⁻¹ suspension. Similarly, the highest concentration of neem yielded the highest egg (27.3%) and nymph mortalities (75.5%), which was also not significantly different from the 0.5% suspension. Therefore, 0.5% neem was used along with 10⁷ B. bassiana conidia ml⁻¹ suspension as an integrated pest management program against B. tabaci . The combination of B. bassiana and neem yielded the highest B. tabaci egg (29.5%) and nymph mortalities (97.2%), and the lowest LT₅₀ (2.08 day) value. Moreover, an integrated combination of B. bassiana with neem caused 27.6 and 20.5% more nymphal mortality than individual treatments of B. bassiana and neem, respectively, 7 days post-application. Thus, a combined application of an entomopathogenic fungus and a botanical insecticide may benefit from both, and it has proven effective for the control of B. tabaci on eggplant.     

Prokaryotic K(+) channels: from crystal structures to diversity

The deep roots and wide branches of the K⁺-channel family are evident from genome surveys and laboratory experimentation. K⁺-channel genes are widespread and found in nearly all the free-living bacteria, archaea and eukarya. The conservation of basic structures and mechanisms such as the K⁺ filter, the gate, and some of the gate's regulatory domains have allowed general insights on animal K⁺ channels to be gained from crystal structures of prokaryotic channels. Since microbes are the great majority of life's diversity, it is not surprising that microbial genomes reveal structural motifs beyond those found in animals. There are open-reading frames that encode K⁺-channel subunits with unconventional filter sequences, or regulatory domains of different sizes and numbers not previously known. Parasitic or symbiotic bacteria tend not to have K⁺ channels, while those showing lifestyle versatility often have more than one K⁺-channel gene. It is speculated that prokaryotic K⁺ channels function to allow adaptation to environmental and metabolic changes, although the actual roles of these channels in prokaryotes are not yet known. Unlike enzymes in basic metabolism, K⁺ channel, though evolved early, appear to play more diverse roles than revealed by animal research. Finding and sorting out these roles will be the goal and challenge of the near future.