Drop-off behaviour of Bt-resistant and Bt-susceptible Helicoverpa armigera (Hübner) (Lepidoptera: Noctuidae) larvae on Bt-cotton and non-Bt cotton plants.

The highest natural mortality rate of larval Lepidoptera in field populations occurs in the first instar, but it is highly variable. The pattern and degree of survival is not easily predicted but depends on their ability to establish on host plants. Lepidopteran larval dispersal behaviour, known as 'drop-off', happens when the host is unsuitable for larvae to settle and begin feeding. Understanding drop-off behaviour of Helicoverpa armigera (Hübner) with and without physiological resistance to Bt toxins on Bt and non-Bt cotton plants is an important component for resistance management strategies for this insect. We examined the drop-off behaviour of H. armigera to determine: (1) whether they move the same way or differently in response to Bt and non-Bt, and (2) could H. armigera larvae detect Bt toxin levels in cotton plants or did they move independently of toxin levels? In this study, we assessed the drop-off behaviour of Bt-resistant and Bt-susceptible H. armigera neonates on artificial diets and cotton plants with and without Bt toxin during the first 12 h after hatching. Bt-resistant and Bt-susceptible H. armigera neonates behaved differently on Bt and non-Bt substrates. The percentages of Bt-resistant larvae that dropped off Bt and non-Bt cotton plants were not significantly different. In contrast, significantly more Bt-susceptible larvae dropped off Bt cotton than non-Bt cotton plants over time. Although Bt-susceptible larvae could not detect Bt toxin, they showed preference on non-Bt toxin substrates and were more likely to drop off substrates with Bt toxin.

Genomic innovations, transcriptional plasticity and gene loss underlying the evolution and divergence of two highly polyphagous and invasive Helicoverpa pest species.

BACKGROUND: Helicoverpa armigera and Helicoverpa zea are major caterpillar pests of Old and New World agriculture, respectively. Both, particularly H. armigera, are extremely polyphagous, and H. armigera has developed resistance to many insecticides. Here we use comparative genomics, transcriptomics and resequencing to elucidate the genetic basis for their properties as pests. RESULTS: We find that, prior to their divergence about 1.5 Mya, the H. armigera/H. zea lineage had accumulated up to more than 100 more members of specific detoxification and digestion gene families and more than 100 extra gustatory receptor genes, compared to other lepidopterans with narrower host ranges. The two genomes remain very similar in gene content and order, but H. armigera is more polymorphic overall, and H. zea has lost several detoxification genes, as well as about 50 gustatory receptor genes. It also lacks certain genes and alleles conferring insecticide resistance found in H. armigera. Non-synonymous sites in the expanded gene families above are rapidly diverging, both between paralogues and between orthologues in the two species. Whole genome transcriptomic analyses of H. armigera larvae show widely divergent responses to different host plants, including responses among many of the duplicated detoxification and digestion genes. CONCLUSIONS: The extreme polyphagy of the two heliothines is associated with extensive amplification and neofunctionalisation of genes involved in host finding and use, coupled with versatile transcriptional responses on different hosts. H. armigera's invasion of the Americas in recent years means that hybridisation could generate populations that are both locally adapted and insecticide resistant.

Oviposition site selection and survival of susceptible and resistant larvae of Helicoverpa armigera (Lepidoptera: Noctuidae) on Bt and non-Bt cotton.

In Australia Bt cotton has been planted since 1996, and has greatly improved the control of its key target Helicoverpa armigera (Hübner). There is no strong evidence that genetically modified cotton has been selected for significant physiological resistance to Bt toxin in field populations. There are many possible explanations for the lack of apparent selection that range from high compliance with the resistance management strategy for this technology to a lack of behavioral preference in key traits such as oviposition that could favor survival. To date most experiments that test oviposition of H. armigera on Bt cotton vs. conventional cotton have been done with susceptible moths. We determine the oviposition preference of a field isolated Bt resistant line of H. armigera and a susceptible counterpart when given a choice of non-Bt cotton and Bt-cotton with the same genetic background, and test whether there is any relationship between oviposition site selection (different plant structures) and the survival of the first instar larvae. Within cotton plants, our experiments consistently showed that both resistant and susceptible moths did not choose plants or plant parts that were less toxic in terms of Bt toxin on which to lay eggs. There was one exception in that susceptible moths were more likely to lay eggs on squares of Bt cotton plants than squares of non-Bt cotton. As expected, the mortality of susceptible H. armigera neonates was significantly higher on structures of Bt cotton plants than on those structures of conventional cotton, and survival was greater on flowers than on other structures of Bt cotton. This confirms opportunities for selection for resistance, and demonstrates no advantage in this respect to carrying resistance genes that might overcome the Bt toxins.

Dual Cry2Ab and Vip3A resistant strains of Helicoverpa armigera and Helicoverpa punctigera (Lepidoptera: Noctuidae); testing linkage between loci and monitoring of allele frequencies.

Considerable attention has been given to delaying the evolution of insect resistance to toxins produced by transgenic crops. The major pests of cotton in Australia are the Lepidoptera Helicoverpa armigera (Hubner, 1805) and Helicoverpa punctigera (Wallengren), and the toxins deployed in current and imminent transgenic cotton varieties are Cry1Ac, Cry2Ab and Vip3A from Bacillus thuringiensis. In this study, lines that carry alleles conferring resistance to Cry2Ab and Vip3A were isolated using F2 tests. Extensive work on the Cry2Ab resistant lines, and preliminary work on the Vip3A resistant lines, suggested a single common resistance to each toxin in both species thereby justifying the use of more efficient F1 tests as the primary means for monitoring changes over time. A potential further efficiency could be gained by developing a single resistant line that carries both types of Bt resistance. Herein we report on work with both H. armigera and H. punctigera that tests whether dual Cry2Ab-Vip3A resistant lines can be developed and, if so, whether they can be used to effectively monitor resistance frequencies. Furthermore, the creation of dual resistant lines allowed linkage between the Cry2Ab and Vip3A resistances to be investigated for H. punctigera. We show that dual resistant lines can be used to increase the efficiency of the F1 screen for recessive alleles, and that in H. punctigera there is no linkage between Cry2Ab and Vip3A resistance.

Geographic variation in antisnake tactics: the evolution of scent-mediated behavior in a lizard.

We used modern comparative methods to examine the evolution of scent-mediated antisnake behavior in the rock-dwelling velvet gecko (Oedura lesueurii). The selective agent is a snake species (broad-headed snake, Hoplocephalius bungaroides) that feeds primarily on velvet geckos by remaining sedentary in rock crevices for days or weeks, waiting to ambush lizards. The past and present distribution of this predator is well documented because of its threatened conservation status. We used this information to sample lizards from three populations distributed with snakes (sympatric) and three populations that appear never to have been distributed with snakes (allopatric) in each of two widespread but geographically distinct genetic groups of velvet gecko (as determined using allozyme electrophoresis). Wild-caught immature geckos from sympatric populations showed higher tongue-flick rates and stronger shifts in locomotion (increased duration of crawling and remaining stationary while pressed against the rock) toward snake-scented rocks than did lizards from allopatric populations. However, predation environment did not significantly affect a lizard's tendency to display other typical antisnake tactics such as tail waving or fleeing. These results were highly repeatable across the two sampled genetic groups of velvet gecko, despite demonstrable genetic divergence between groups. Experiments with hatchling lizards that had no experience with predators indicate that qualitative components of antisnake behaviors are probably inherited. The method of phylogenetically independent contrasts strongly suggests that the presence or absence of snakes has driven the evolution of behavior in velvet geckos. Collectively, these results provide support for an often suggested but speculative expectation that prey can adapt to predation pressure on a local scale.

The use of the selective serotonin S2 receptor antagonist Ketanserin in the treatment of Raynaud's phenomenon.

Ketanserin, a selective S2-receptor antagonist was evaluated in a double-blind parallel placebo-controlled trial of 29 patients with Raynaud's phenomenon. The effect on finger blood flow was assessed by strain-gauge plethysmography during a standardised cooling stress. Ten normal healthy controls also had finger blood flow assessments made in the same way. The Ketanserin group showed a significantly greater improvement (P less than 0.05) in finger blood flow at minimum flow temperature than the placebo group. The results suggest that the local release of serotonin may play a role in Raynaud's phenomenon and that Ketanserin can modify the abnormal response in finger blood flow.

Diagnosis and successful treatment of subacute erysipelas in a captive dolphin.

Ersipelas, a common disease of swine, is caused by Erysipelothrix rhusiopathiae. The organism was isolated in a blood culture taken from an infected captive dolphin. The dolphin showed typical subacute symptoms of square- and diamond-shaped skin lesions as seen in swine. It was surmised, in retrospect, that the disease was secondary to a primary pneumonia. The symptoms, clinical pathology and other special examinations, treatment and response are discussed.

Anesthetization of a Cape fur seal (Arctocephalus pusillus) for the treatment of a chronic eye infection and amputation of a metatarsal bone.

Tranquilization using 3 mg/kg of ketamine by intramuscular injection followed by inhalation anaesthesia using halothane was performed on an adult Cape fur seal in order to perform a metatarsal amputation and ophthalmic examination. Ketamine was found to have little effect at the dosage used while halothane proved to be a rapid induction agent providing a safe, continued level of surgical anaesthesia. Variations in cardiac rate and body temperature were recorded during anesthasia and blood was sampled for haematology.