A draft Diabrotica virgifera virgifera genome: insights into control and host plant adaption by a major maize pest insect.

Background: Adaptations by arthropod pests to host plant defenses of crops determine their impacts on agricultural production. The larval host range of western corn rootworm, Diabrotica virgifera virgifera (Coleoptera: Chrysomelidae), is restricted to maize and a few grasses. Resistance of D.

Comparative transcriptomics of ice-crawlers demonstrates cold specialization constrains niche evolution in a relict lineage.

Key changes in ecological niche space are often critical to understanding how lineages diversify during adaptive radiations. However, the converse, or understanding why some lineages are depauperate and relictual, is more challenging, as many factors may constrain niche evolution. In the case of the insect order Grylloblattodea, highly conserved thermal breadth is assumed to be closely tied to their relictual status, but has not been formerly tested.

The evolutionary dynamics of variant antigen genes in Babesia reveal a history of genomic innovation underlying host-parasite interaction.

Babesia spp. are tick-borne, intraerythrocytic hemoparasites that use antigenic variation to resist host immunity, through sequential modification of the parasite-derived variant erythrocyte surface antigen (VESA) expressed on the infected red blood cell surface. We identified the genomic processes driving antigenic diversity in genes encoding VESA (ves1) through comparative analysis within and between three Babesia species, (B.

Experimental vaccination of sheep and cattle against tick infestation using recombinant 5'-nucleotidase.

Limited prior evidence suggests that 5'-nucleotidase, an ectoenzyme principally located in the Malpighian tubules of the tick Rhipicephalus (Boophilus) microplus, could be an effective antigen in an anti-tick vaccine. To assess this, recombinant 5'-nucleotidase was expressed in Escherichia coli and used in vaccination trials with both sheep and cattle. Vaccinated sheep were challenged with freshly moulted adult ticks.

Antigen cocktails: valid hypothesis or unsubstantiated hope?

After more than three decades of research into the development of vaccines against parasites, a substantial number of antigens have been identified that, as purified native proteins or recombinant proteins, induce some protection against the target parasite. Very few achieve a degree of efficacy likely to make them candidates for single-antigen vaccines. Therefore, multi-antigen or 'cocktail' vaccines are proposed based on the assumption that such cocktails will show enhanced efficacy.

Acaricide rotation strategy for managing resistance in the tick Rhipicephalus (Boophilus) microplus (Acarina: Ixodidae): laboratory experiment with a field strain from Costa Rica.

During the past two decades, resistance to pyrethroids within the cattle tick genus Boophilus has caused tick control problems in various tropical countries, mainly in Latin America, southern Africa, Australia, and New Caledonia. A Rhipicephalus (Boophilus) microplus (Canestrini) strain from Costa Rica, exhibiting resistance to the pyrethroid deltamethrin but only a very low resistance to organophosphates (OP) was kept under selection pressure for 9 to 11 generations by using deltamethrin or coumaphos (OP), either exclusively or in rotation. The objective of this acaricide rotation was to examine the possibility of delaying or reducing the full emergence of pyrethroid resistance.

A ten-year review of commercial vaccine performance for control of tick infestations on cattle.

Ticks are important ectoparasites of domestic and wild animals, and tick infestations economically impact cattle production worldwide. Control of cattle tick infestations has been primarily by application of acaricides which has resulted in selection of resistant ticks and environmental pollution. Herein we discuss data from tick vaccine application in Australia, Cuba, Mexico and other Latin American countries.

Advances in the genomics of ticks and tick-borne pathogens.

Ticks and the diseases for which they are vectors engage in complex interactions with their mammalian hosts. These interactions involve the developmental processes of tick and pathogen, and interplay between the defensive responses and counter responses of host, tick and pathogen. Understanding these interactions has long been an intractable problem, but progress is now being made thanks to the flood of genomic information on host, tick and pathogen, and the attendant, novel experimental tools that have been generated.

Control of parasitic disease using vaccines: an answer to drug resistance?

Antiparasitic drugs have been used successfully to control parasitic diseases in animals for many years, as they are safe, cheap and effective against a broad spectrum of parasites. One drawback of this success appears to be the emergence of drug resistance in many target parasites. Moreover, issues of residues in the food chain and environment have arisen, which threaten their sustained use.

Vaccination against ectoparasites.

Ectoparasites of livestock are of great economic and social importance but their effective control remains difficult. The feasibility of vaccination as a novel control measure was established over a decade ago with the commercial release of a recombinant vaccine against the cattle tick Boophilus microplus. Since then, research has continued on ticks and other ectoparasites.

Sequencing a new target genome: the Boophilus microplus (Acari: Ixodidae) genome project.

The southern cattle tick, Boophilus microplus (Canestrini), causes annual economic losses in the hundreds of millions of dollars to cattle producers throughout the world, and ranks as the most economically important tick from a global perspective. Control failures attributable to the development of pesticide resistance have become commonplace, and novel control technologies are needed. The availability of the genome sequence will facilitate the development of these new technologies, and we are proposing sequencing to a 4-6X draft coverage.

Tick control: thoughts on a research agenda.

Tick control is critical to the control of tick borne disease, while the direct impact of ticks on livestock productivity is also well known. For livestock, tick control today rests overwhelmingly on the twin approaches of genetics and chemical acaricides, although the disadvantages and limitations of both are recognized. The achievement of the full potential of vaccination, the application of biocontrol agents and the coordinated management of the existing technologies all pose challenging research problems.

Potential for recombinant Babesia bovis antigens to protect against a highly virulent isolate.

Two antigens from Babesia bovis,12D3 and 11C5, were expressed and purified as recombinant proteins in Escherichia coli and used to vaccinate groups of six Babesia-susceptible cattle. These were subsequently challenged with a highly virulent strain of B. bovis.

Anti-tick vaccines.

There is now abundant evidence that vaccination with defined protein antigens is able to induce significant immunity to tick infestation. In a limited number of cases, this immunity has been duplicated by vaccination with recombinant antigens, a critical step on the pathway to commercial vaccine production. The existence of two commercial vaccines has allowed a number of field studies showing that the existing products can make an important contribution to an integrated approach to the control of ticks in the field.

Veterinary parasitic vaccines: pitfalls and future directions.

Most available antiparasitic drugs are safe, cheap and highly effective against a broad spectrum of parasites. However, the alarming increase in the number of parasite species that are resistant to these drugs, the issue of residues in the food chain and the lack of new drugs stimulate development of alternative control methods in which vaccines would have a central role. Parasite vaccines are still rare, but there are encouraging signs that their number will increase in the next decade.

A novel antigen from Anaplasma marginale: characterization, expression and preliminary evaluation of the recombinant protein.

Through a process of protein fractionation and vaccination we previously identified four native antigens that confer a degree of protection against challenge with Anaplasma marginale. One of these, Ana 29 has been successfully cloned and sequenced using degenerate primers designed to N-terminal and internal peptide sequences. The full-length gene codes for a protein with a theoretical molecular weight of 27 kDa and pI 8.

Identification of novel protective antigens from Anaplasma marginale.

A successful recombinant vaccine against Anaplasma marginale remains elusive, despite intensive study of the protective, though variable major surface antigens. As an alternative approach to the discovery of additional antigens, crude parasite material was subjected to conventional protein fractionation, coupled with vaccination and parasite challenge trials, without making assumptions as to the nature or location of these antigens. This has lead to the identification of four novel antigens that, in microgram amounts, have significant protective effects in vaccination trials.

The purine salvage enzyme hypoxanthine guanine xanthine phosphoribosyl transferase is a major target antigen for cell-mediated immunity to malaria.

Although there is good evidence that immunity to the blood stages of malaria parasites can be mediated by different effector components of the adaptive immune system, target antigens for a principal component, effector CD4(+) T cells, have never been defined. We generated CD4(+) T cell lines to fractions of native antigens from the blood stages of the rodent parasite, Plasmodium yoelii, and identified fraction-specific T cells that had a Th1 phenotype (producing IL-2, IFN-gamma, and tumor necrosis factor-alpha, but not IL-4, after antigenic stimulation). These T cells could inhibit parasite growth in recipient severe combined immunodeficient mice.

Reduced oviposition of Boophilus microplus feeding on sheep vaccinated with vitellin.

The most abundant protein present in Boophilus microplus eggs, vitellin, was isolated and purified as a non-covalent complex of six glyco-polypeptides of Mr 44-107kDa. The protein complex bound haem. Immuno-blots demonstrated that antibodies raised to vitellin recognised a 200kDa polypeptide in the haemolymph of adult female ticks.

The molecular revolution in the development of vaccines against ectoparasites.

Over the last decade, the application of a spectrum of molecular techniques has begun to revolutionise our understanding of protective immune responses to ectoparasites and the targets for those responses. The catalogue of potential and actual protective antigens characterised in detail is slowly expanding. The validity of regarding such antigens as generic and capable of cross-species protection is being explored.

Evidence for the utility of the Bm86 antigen from Boophilus microplus in vaccination against other tick species.

The Bm86 antigen, as originally identified in Boophilus microplus, is the basis of commercial tick vaccines against this tick species. The potential for using this antigen or homologues of the antigen in vaccination against other tick species has been assessed. We have conducted vaccine trials in cattle using the B.

Biochemical and molecular characterization of serine proteases from larvae of Chrysomya bezziana, the Old World Screwworm fly.

The diversity of serine proteases secreted from Chrysomya bezziana larvae was investigated biochemically and by PCR and sequence analysis. Cation-exchange chromatography of purified larval serine proteases resolved four trypsin-like activities and three chymotrypsin-like activities as discerned by kinetic studies with benzoyl-Arg-p-nitroanilide and succinyl-Ala-Ala-Pro-Phe-p-nitroanilide. Amino-terminal sequencing of the three most abundant fractions gave two sequences, which were homologous to other Dipteran trypsins and chymotrypsins.

Identification and molecular characterisation of a peritrophin gene, peritrophin-48, from the myiasis fly Chrysomya bezziana.

The peritrophic matrix lines the midgut of most insects and has important roles in digestion, protection of the midgut from mechanical damage and invasion by micro-organisms. Although a few intrinsic peritrophic matrix proteins have been characterised, no direct homologues of any of these proteins have been found in other insect species, even closely related species, suggesting that the peritrophic matrix proteins show considerable sequence divergence. We now report the identification of the cDNA and genomic DNA sequences of a Chrysomya bezziana homologue of the Lucilia cuprina intrinsic peritrophic matrix protein, peritrophin-48.

A novel family of chitin-binding proteins from insect type 2 peritrophic matrix. cDNA sequences, chitin binding activity, and cellular localization.

The peritrophic matrix is a prominent feature of the digestive tract of most insects, but its function, formation, and even its composition remain contentious. This matrix is a molecular sieve whose toughness and elasticity are generated by glycoproteins, proteoglycans, and chitin fibrils. We now describe a small, highly conserved protein, peritrophin-15, which is an abundant component of the larval peritrophic matrices of the Old World screwworm fly, Chrysomya bezziana, and sheep blowfly, Lucilia cuprina.

Vaccination against the Old World screwworm fly (Chrysomya bezziana).

Chrysomya bezziana is an endemic pest of livestock or a threat to livestock production in large areas of Africa, the Middle East, southern and south-east Asia and Australia. Its control is difficult. The feasibility of vaccinating against this pest has now been explored.