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Laboratory evaluation of controlled-release insect repellent formulations.

Seven microcapsule formulations and two polymer formulations of deet were tested on white rabbits for their repellency against the mosquito, Aedes aegypti. Two microcapsule formulations and one polymer formulation provided more than 80% protection for 12 hours. Results demonstrated that the protection period of deet can be extended through controlled-release techniques.

Administration, Cutaneous

Risk factors associated with Corynebacterium pseudotuberculosis infection in California horses.

A case-control study was designed using equine medical records from the UC Davis Veterinary Medical Teaching Hospital (VMTH) and data derived through a mailed survey. The objective was to evaluate the associations between horse demographics, horse-management factors, and equine Corynebacterium pseudotuberculosis infection in California. Horses admitted to the VMTH between July 1 1992 and June 30 1994 served as the study base for case identification and simple random sampling of 800 controls. A questionnaire was mailed to the owners of all horses enrolled in the study to collect data on demographics, management and health-related questions. A logistic-regression model containing age, outdoor activity level, other locations in California, insect-control measures, contact with other horses, and summer pasture was developed. The final model was adjusted for the suspected confounding variables admission type, regular teaching hospital patient and breed. Horses of age between 1 and 2 yrs and between 3 and 5 yrs, and horses in contact with other horses or horses on summer pasture had significantly increased odds (p < 0.05) of being diagnosed with C. pseudotuberculosis infection. The results support the hypotheses that the disease predominantly affects young-adult horses of all breeds and both sexes, and that management factors play an important role in occurrence of the disease. Since the existing serological test system is not reliable and destruction of infected animals is not feasible, the most-logical approach for disease prevention is the early identification and isolation of clinical cases and the implementation of management changes like improvement of stable hygiene and insect control and change of pasture practices.

Animals

A bifunctional heterodimeric insect neuropeptide analog.

The synthesis and biological activity of a bifunctional, heterodimeric insect neuropeptide analog are described. The heterodimer is composed of the C-terminal pentapeptide active core regions of the leucokinin/achetakinin and pyrokinin neuropeptide families linked via their N-terminal amino groups with a succinyl diacid moiety. Members of the leucokinin/achetakinin family can induce fluid secretion in malpighian tubules of the house cricket, Acheta domesticus, whereas the pyrokinins demonstrate activity in a cricket oviduct myotropic bioassay. No cross-activity is observed for the two neuropeptide families in these bioassays. However, the heterodimer elicits responses in both Acheta bioassays. Such a bifunctional analog may in future serve as a template for the design of stable, bifunctional pest insect control agents of greater efficiency.

Amino Acid Sequence

Male-linked translocations and the control of insect pest populations.

Published data on an experimental release of Culex pipiens carrying a male-linked translocation are re-examined and it is shown that the steady decline in translocation frequency after termination of releases agrees with theoretical expectations, because of the selective disadvantage of translocation heterozygote males. Systems based on negative heterosis or meiotic drive are considered whereby it might be possible to prolong the population control which would be achieved by a short term release.

Animals

Insect neuropeptides: discovery and application in insect management.

New approaches to the development of insect control agents have been revealed through the molecular description of neuropeptides, their biogenesis, action, and degradation. Prerequisite to the exploitation of a neuropeptide as a lead to control agent development is a thorough understanding of the biochemistry of the neuropeptide and appreciation of its physiological impact. Reliable bioassays must be coupled with advanced biochemical and molecular genetic technologies to overcome limitations imposed by the typically low endogenous levels of individual neuropeptides. Purification, amino acid sequencing, and gene cloning provide the molecular tools necessary for studies on neuropeptide synthesis, processing, secretion, receptor binding, and inactivation. Each of these areas consists of a number of amino acid sequence-, and enzyme-dependent steps which may be considered as targets for the development of highly specific control agents. These agents will include antagonist and superagonists, peptidomimetics, recombinant peptides delivered through the baculovirus technology, receptor blockers, and enzyme inhibitors.

Amino Acid Sequence

Evolution and structural conservation of the control region of insect mitochondrial DNA.

The control regions of mitochondrial DNA of two insects, Schistocerca gregaria and Chorthippus parallelus, have been isolated and sequenced. Their sizes are 752 bp and 1,512 bp, respectively, with the presence of a tandem repeat in C. parallelus. (The sequences of the two repeats are highly conserved, having a homology of 97.5%.) Comparison of their nucleotide sequences revealed the presence of several conserved sequence blocks dispersed through the whole control region, showing a different evolutionary pattern of this region in these insects as compared to that in Drosophila. A highly conserved secondary structure, located in the 3' region near the small rRNA gene, has been identified. Sequences immediately flanking this hairpin structure rather than the sequences of this structure themselves are conserved between S. gregaria/C. parallelus and Drosophila, having a sequence consensus of "TATA" at 5' and "GAA(A)T" at 3'. The motif "G(A)nT" is also present in the 3' flanking sequences of mammalian, amphibian, and fish mitochondrial L-strand replication origins and a potential plant mitochondrial second-strand-replication origin, indicating its universal conservation and functional importance related to replication origins. The stem-and-loop structure in S. gregaria/C. parallelus appears to be closely related to that found in Drosophila despite occupying a different position, and may be potentially associated with a second-strand-replication origin. This in turn suggests that such a secondary structure might be widely conserved across invertebrates while their location in the control region may be variable. We have looked for such a conserved structure in the control regions of two other insects, G. firmus and A. mellifera, whose DNA sequences have been published, and their possible presence is discussed. Mitochondrial control regions characterized to date in five different insect taxa (Drosophila, G. firmus, A. mellifera, S. gregaria, and C. parallelus) may be classed into two distinct groups having different evolutionary patterns. It is observed that tandem repetition of regions containing a probable replication origin occurred in some species from disjunct lineages in both groups, which would be the result of convergent evolution. We also discuss the possibility of a mechanism of "parahomologous recombination by unequal crossing-over" in mitochondria, which can explain the generation of such tandemly repeated sequences (especially the first critical repetition) in the control region of mtDNA, and also their convergent evolution in disjunct biological lineages during evolution.

Animals

Effects of red-headed in sawfly, Neodiprion lecontei, nuclear polyhedrosis virus on rainbow trout, Salmo gairdneri, and Daphnia pulex.

The effects of a nuclear polyhedrosis virus (NPV) of the red-headed pine sawfly, Neodiprion lecontei, on rainbow trout, Salmo gairdneri, were investigated. The fish were exposed to this virus by intubation and topical application and no ill-effects were observed. Similarly, no ill-effects were detected in Daphnia pulex when the same NPV was added to their culture medium. The materials were lyophilized, NPV-infected sawfly larvae (normally used for insect control), lyophilized, uninfected larvae and purified, polyhedral inclusion bodies. On the basis of these laboratory tests, this virus, when disseminated as a biocontrol agent, should present no hazard to rainbow trout or to the aquatic invertebrate Daphnia pulex, two species frequently used in toxicity tests of chemical pesticides.

Animals

VSG gene expression site control in insect form Trypanosoma brucei.

When the African trypanosome Trypanosoma brucei is taken up from mammals by a tse-tse fly, it replaces its variant surface glycoprotein (VSG) coat by a procyclin coat. Transcription of VSG genes stops in the fly, but transcription of sequences derived from the promoter area of the VSG expression site(s) remains high. Whether this is due to continuing high activity of one promoter or to low activity of many promoters was unclear. We have used the small differences between the sequences of different expression sites to show that multiple expression site promoters are active in insect form trypanosomes. This is confirmed by the low expression of single copy marker genes introduced into the transcribed area. However, if the expression site promoter is removed from the genomic location of the expression site and inserted in the non-transcribed spacer of the ribosomal DNA (rDNA), it is derepressed. Derepression of transcription can also be accomplished by replacing the promoter of an expression site by an rDNA promoter. We conclude that the down-regulation of VSG gene expression site promoters in insect form trypanosomes is affected by both the DNA sequence of the promoter and the genomic context in which it resides.

Animals

Microtubules and control of insect egg shape.

This study evidence for tension transmission by microtubules and desmosomes in the follicular epithelium during anisometric growth of certain insect eggs. Most insect oocytes, and the follicles which surround them, grow anisometrically as they assume shapes which approximate to those of long prolate spheroids. Surface growth is most rapid in directions which parallel the polar axis of an oocyte and slowest in circumferential directions at right angles to this axis. The longitudinal axes of microtubule bundles in follicle cells of the gall midge Heteropeza and the cockroach Periplaneta are oriented circumferentially with respect to the surfaces of developing eggs and at right angles to the polar axes of eggs. At cell boundaries, the tubules appear to be attached to spot desmosomes. It is suggested that microtubules and desmosomes form a mechanical continuum throughout a follicular epithelium which transmits tensile forces around the circumference of a growing egg. Follicular resistance to circumferential expansion may be largely responsible for defining the elongate form of insect eggs.

Animals

Control of insect ecdysis by a positive-feedback endocrine system: roles of eclosion hormone and ecdysis triggering hormone.

A successful ecdysis in insects requires the precise coordination of behaviour with the developmental changes that occur late in a moult. This coordination involves two sets of endocrine cells: the peripherally located Inka cells, which release ecdysis triggering hormone (ETH), and the centrally located neurosecretory neurones, the VM neurones, which release eclosion hormone (EH). These two sets of endocrine cells mutually excite one another: EH acts on the Inka cells to cause the release of ETH. ETH, in turn, acts on the VM neurones to cause the release of EH. This positive-feedback relationship allows the Inka cells and the VM neurones to be the peripheral and central halves, respectively, of a decision-making circuit. Once conditions for both halves have been satisfied, their reciprocal excitation results in a massive EH/ETH surge in the blood as well as a release of EH within the central nervous system. This phasic signal then causes the tonic activation of a distributed network of peptidergic neurones that contain crustacean cardioactive peptide. The relationship of the latter cells to the subsequent maintenance of the ecdysis motor programme is discussed.

Animals