Development of resistance to nymphs of Amblyomma cajennense ticks (Acari : Ixodidae) in dogs

Ticks have long been regarded as constraints to humans and domestic animals, but hosts often develop resistance to ticks after repeated infestations. The purpose of this investigation was to study the possible acquisition of immunity in domestic dogs to nymphs of A. cajennense by determining the tick alimentary performance after successive controlled infestations. Mean engorged weight of nymphs was not significantly different among the three infestations; molting rate from nymph to adult ticks, and the percentage of nymph recovery were also very close in all infestations. These results are similar to those obtained in studies of the dog-adult Rhipicephalus sanguineus interface. It is concluded that domestic dogs do not develop resistance against nymphs of A. cajennense ticks.

Allergy to hematophagous arthropods bites

Allergies to hematophagous arthropod bites are inflammatory reactivity to arthropods salivary components. They vary in intensity and quality dependent upon the arthropod species and the individual immune response to specific proteins of the insect s saliva. Individuals who were not previously exposed show mild localized reactions not beyond those expected by pharmacological substances present in arthropods saliva. Allergic reactions are immunological in their nature and the diversity derived from hypersensitivity reactions with different levels of participation of the immune system components. Some are mainly derived from a humoral immune response, and others are based predominantly on T-lymphocyte-mediated. The majority of these undesired biological answers are self-limited, and few may cause a systemic reaction. This article intends to discuss the immunological ingredients of this evolutionary interaction.

Efeito do gradiente interior-borda-matriz, e do tipo de matriz sobre a entomofaúna edáfica em fragmentos de cerrado

edge effect. Thus, under the influence of the adjacent matrix, fragments undergo microclimatic alterations that accentuate changes in species composition and community structure. In order to better understand edge and matrix effects on the richness and abundance of edaphic arthropods, this study assessed: (a) the difference between habitat (fragment) and non-habitat (matrix); (b) whether there is a continuous interior-edge-matrix gradient; and (c) the difference between matrices for arthropod orders richness and abundance. We selected 15 landscapes, 5 of which contained a cerrado fragment surrounded by sugarcane cultivation, 5 with a cerrado fragment within eucalyptus and 5 with a cerrado fragment within pasture. In each landscape the soil fauna was collected along with the soil and then extracted with the aid of the modified Berlese-Tullgren funnel. We chose the orders Coleoptera, Collembola, Mesostigmata and Oribatida for analysis, and after separation of the individuals we used model selection analysis via AIC. The model type fragment x matrix was the most likely to explain richness, total and relative abundances of the four orders (wAICc between 0,6623 and 1,0). The model of edge distance (edge effect) was plausible to total abundance and relative abundance of Mesostigmata order (wAICc=0,2717 and 0,186). Local environmental variables (soil texture, temperature and relative humidity), and fragment size were also measured to avoid confounding factors and were not presented as plausible models to explain the patterns. So edaphic arthropods, despite protecting themselves under the ground, are extremely sensitive to fragmentation, even with the replacement of natural habitat by agricultural use, such as sugarcane, pasture and eucalyptus. This group should be studied environmental impact assessments because provides important ecosystem se ravincde s inacnludd eisd ainn efficient bio-indicator

Report of the Poplar Working Group

Poplars consist of all species of the genus Populus, including cottonwoods, aspens, and the many interspecies hybrids in common use (Dickmann and Stuart 1983). Our working group focused on the fungal pathogens, arthropod herbivores, and weed competitors of Populus in the United States. However, bacterial and viral diseases of Populus are significant in Europe, and genetic engineering approaches toward their control or management are being studied. The key aspects of poplar biology important to understanding the use of pest resistance genes are described below.

An Investigation of Wild Bee Diversity and Abundance in Plots Managed by The Nature Conservancy in South-Central Nebraska and of Beneficial Arthropods Associated with Native Nebraska Flora

Insect pollination is an essential ecosystem service, and bees are the principal pollinators of wild and cultivated plants. Habitat management and enhancement are a proven way to encourage wild bee populations, providing them with food and nesting resources. I examined bee diversity and abundance in plots managed by The Nature Conservancy near Wood River, NE. The plots were seeded with 2 seed mixes at 2 seeding rates: high diversity mix at the recommended rate, high diversity mix double the recommended rate, Natural Resources Conservation Service (NRCS) conservation planting (CP) 25 mix at one-half the recommended rate, and NRCS CP25 mix at the recommended rate. I measured wild bee abundance and diversity, and established a database of wild bees associated with the plots. I also compared genus richness and abundance among the plots using and aerial net and blue vane traps to collect bees. Significant differences were not observed in genus richness and diversity among the plots; however, plot size and the ability of blue vane traps to draw bees from a long distance may have influenced my results. In 2008, 15 genera and 95 individual bees were collected using an aerial net and in 2009, 32 genera and 6,103 individual bees were collected using blue vane traps. I also studied the beneficial insects associated with native Nebraska flora. Seventeen species of native, perennial flora were established in 3 separate plots located in eastern Nebraska. I transplanted four plants of each species in randomized 0.61 m x 0.61 m squares of a 3.05 m x 9.14 m plot. Arthropods were sampled using a modified leaf blower/vacuum. Insects and other arthropods were identified to family and organized into groups of predators, parasites, pollinators, herbivores, and miscellaneous. Associations between plant species and families of beneficial arthropods (predators, parasites, and pollinators) were made. Pycnanthemum flexuosum Walter attracted significantly more beneficial arthropod families than 7 other species of plants tested. Dalea purpurea Vent and Liatris punctata Hook also attracted significantly fewer beneficial arthropod families than 4 other species of plants tested. In total, 31 predator, 11 parasitic, 4 pollinator, 31 herbivore, and 10 miscellaneous families of arthropods were recorded.

Efeito do gradiente interior-borda-matriz, e do tipo de matriz sobre a entomofaúna edáfica em fragmentos de cerrado

edge effect. Thus, under the influence of the adjacent matrix, fragments undergo microclimatic alterations that accentuate changes in species composition and community structure. In order to better understand edge and matrix effects on the richness and abundance of edaphic arthropods, this study assessed: (a) the difference between habitat (fragment) and non-habitat (matrix); (b) whether there is a continuous interior-edge-matrix gradient; and (c) the difference between matrices for arthropod orders richness and abundance. We selected 15 landscapes, 5 of which contained a cerrado fragment surrounded by sugarcane cultivation, 5 with a cerrado fragment within eucalyptus and 5 with a cerrado fragment within pasture. In each landscape the soil fauna was collected along with the soil and then extracted with the aid of the modified Berlese-Tullgren funnel. We chose the orders Coleoptera, Collembola, Mesostigmata and Oribatida for analysis, and after separation of the individuals we used model selection analysis via AIC. The model type fragment x matrix was the most likely to explain richness, total and relative abundances of the four orders (wAICc between 0,6623 and 1,0). The model of edge distance (edge effect) was plausible to total abundance and relative abundance of Mesostigmata order (wAICc=0,2717 and 0,186). Local environmental variables (soil texture, temperature and relative humidity), and fragment size were also measured to avoid confounding factors and were not presented as plausible models to explain the patterns. So edaphic arthropods, despite protecting themselves under the ground, are extremely sensitive to fragmentation, even with the replacement of natural habitat by agricultural use, such as sugarcane, pasture and eucalyptus. This group should be studied environmental impact assessments because provides important ecosystem se ravincde s inacnludd eisd ainn efficient bio-indicator

Abiotic factors shape temporal variation in the structure of an ant-plant network

Despite recognition of key biotic processes in shaping the structure of biological communities, few empirical studies have explored the influences of abiotic factors on the structural properties of mutualistic networks. We tested whether temperature and precipitation contribute to temporal variation in the nestedness of mutualistic ant-plant networks. While maintaining their nested structure, nestedness increased with mean monthly precipitation and, particularly, with monthly temperature. Moreover, some species changed their role in network structure, shifting from peripheral to core species within the nested network. We could summarize that abiotic factors affect plant species in the vegetation (e.g., phenology), meaning presence/absence of food sources, consequently an increase/decrease of associations with ants, and finally, these variations to fluctuations in nestedness. While biotic factors are certainly important, greater attention needs to be given to abiotic factors as underlying determinants of the structures of ecological networks.

Lufaxin, a Novel Factor Xa Inhibitor From the Salivary Gland of the Sand Fly Lutzomyia longipalpis Blocks Protease-Activated Receptor 2 Activation and Inhibits Inflammation and Thrombosis In Vivo

Objective-Blood-sucking arthropods' salivary glands contain a remarkable diversity of antihemostatics. The aim of the present study was to identify the unique salivary anticoagulant of the sand fly Lutzomyia longipalpis, which remained elusive for decades. Methods and Results-Several L. longipalpis salivary proteins were expressed in human embryonic kidney 293 cells and screened for inhibition of blood coagulation. A novel 32.4-kDa molecule, named Lufaxin, was identified as a slow, tight, noncompetitive, and reversible inhibitor of factor Xa (FXa). Notably, Lufaxin's primary sequence does not share similarity to any physiological or salivary inhibitors of coagulation reported to date. Lufaxin is specific for FXa and does not interact with FX, Dansyl-Glu-Gly-Arg-FXa, or 15 other enzymes. In addition, Lufaxin blocks prothrombinase and increases both prothrombin time and activated partial thromboplastin time. Surface plasmon resonance experiments revealed that FXa binds Lufaxin with an equilibrium constant approximate to 3 nM, and isothermal titration calorimetry determined a stoichiometry of 1:1. Lufaxin also prevents protease-activated receptor 2 activation by FXa in the MDA-MB-231 cell line and abrogates edema formation triggered by injection of FXa in the paw of mice. Moreover, Lufaxin prevents FeCl3-induced carotid artery thrombus formation and prolongs activated partial thromboplastin time ex vivo, implying that it works as an anticoagulant in vivo. Finally, salivary gland of sand flies was found to inhibit FXa and to interact with the enzyme. Conclusion-Lufaxin belongs to a novel family of slow-tight FXa inhibitors, which display antithrombotic and anti-inflammatory activities. It is a useful tool to understand FXa structural features and its role in prohemostatic and proinflammatory events. (Arterioscler Thromb Vasc Biol. 2012;32:2185-2196.)

Phylogeography of Rift Valley Fever Virus in Africa Reveals Multiple Introductions in Senegal and Mauritania

Rift Valley Fever (RVF) virus (Family Bunyaviridae) is an arthropod-borne RNA virus that infects primarily domestic ruminants and occasionally humans. RVF epizootics are characterized by numerous abortions and mortality among young animals. In humans, the illness is usually characterized by a mild self-limited febrile illness, which could progress to more serious complications. RVF virus is widespread and endemic in many regions of Africa. In Western Africa, several outbreaks have been reported since 1987 when the first major one occurred at the frontier of Senegal and Mauritania. Aiming to evaluate the spreading and molecular epidemiology in these countries, RVFV isolates from 1944 to 2008 obtained from 18 localities in Senegal and Mauritania and 15 other countries were investigated. Our results suggest that a more intense viral activity possibly took place during the last century compared to the recent past and that at least 5 introductions of RVFV took place in Senegal and Mauritania from distant African regions. Moreover, Barkedji in Senegal was possibly a hub associated with the three distinct entries of RVFV in West Africa.

Habitat structure and food resources for wildlife across successional stages in a tropical forest

Tropical forests are experiencing an increase in the proportion of secondary forests as a result of the balance between the widespread harvesting of old-growth forests and the regeneration in abandoned areas. The impacts of such a process on biodiversity are poorly known and intensely debated. Recent reviews and multi-taxa studies indicate that species replacement in wildlife assemblages is a consistent pattern, sometimes stronger than changes in diversity, with a replacement from habitat generalists to old-growth specialists being commonly observed during tropical forest regeneration. However, the ecological drivers of such compositional changes are rarely investigated, despite its importance in assessing the conservation value of secondary forests, and to support and guide management techniques for restoration. By sampling 28 sites in a continuous Atlantic forest area in Southeastern Brazil, we assessed how important aspects of habitat structure and food resources for wildlife change across successional stages, and point out hypotheses on the implications of these changes for wildlife recovery. Old-growth areas presented a more complex structure at ground level (deeper leaf litter, and higher woody debris volume) and higher fruit availability from an understorey palm, whereas vegetation connectivity, ground-dwelling arthropod biomass, and total fruit availability were higher in earlier successional stages. From these results we hypothetize that generalist species adapted to fast population growth in resource-rich environments should proliferate and dominate earlier successional stages, while species with higher competitive ability in resource-limited environments, or those that depend on resources such as palm fruits, on higher complexity at the ground level, or on open space for flying, should dominate older-growth forests. Since the identification of the drivers of wildlife recovery is crucial for restoration strategies, it is important that future work test and further develop the proposed hypotheses. We also found structural and functional differences between old-growth forests and secondary forests with more than 80 years of regeneration, suggesting that restoration strategies may be crucial to recover structural and functional aspects expected to be important for wildlife in much altered ecosystems, such as the Brazilian Atlantic forest. (C) 2012 Elsevier B.V. All rights reserved.

Purification and characterization of Hb 98-114: A novel hemoglobin-derived antimicrobial peptide from the midgut of Rhipicephalus (Boophilus) microplus

The antimicrobial activity of hemoglobin fragments (hemocidins) has been reported in a variety of models. The cattle tick Rhipicephalus (Boophilus) microplus is a blood sucking arthropod from where the first in vivo-generated hemocidin was characterized (Hb 33-61). In the present work we identified a novel antimicrobial peptide from the midgut of fully engorged R. (B.) microplus females, which comprises the amino acids 98-114 of the alpha subunit of bovine hemoglobin, and was designated Hb 98-114. This peptide was active against several yeast and filamentous fungi, although no activity was detected against bacteria up to 50 mu M of the synthetic peptide. Hb 98-114 was capable of permeabilizing Candida albicans cell membrane and had a fungicidal effect against this yeast. Circulardichroism (CD) and nuclear magnetic resonance (NMR) experiments showed that Hb 98-114 has a random conformation in aqueous solution but switches to an alpha-helical conformation in the presence of sodium dodecyl sulfate (SDS). This alpha helix adopts an amphipathic structure which may be the mechanism of cell membrane permeabilization. Importantly, Hb 98-114 may play an important role in defending the tick midgut against fungal pathogens and is the first hemocidin with specific antifungal activity to be characterized. (C) 2012 Elsevier Inc. All rights reserved.

Morphological description of the mouthparts of the Asian citrus psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae)

Scanning (SEM) and transmission (TEM) electron microscopy were used to elucidate the morphology of the rostrum, as well as the mandibular and maxillary stylets of the psyllid Diaphorina cirri, vector of phloem-inhabiting bacteria associated with citrus huanglongbing (HLB) disease. D. cirri has a cone-shaped rostrum that extends behind the pair of prothoracic coxae. The stylet bundle comprises a pair of mandibular (Md) and maxillary (Mx) stylets with a mean length of 513.3 mu m; when retracted, their proximal portions form a loop and are stored in the crumena (Cr). Serial cross-sections of the rostrum revealed that the mandibles are always projected in front of the maxillary stylets. The two maxillary stylets form the food and salivary canals, with diameters of 0.9 mu m and 0.4 mu m respectively. These two canals merge at the end of the stylets forming a common duct with a length of 4.3 mu m and a mean diameter of 0.9 mu m. The acrostyle, a distinct anatomical structure present in the common duct of aphid maxillary stylets, was not observed by TEM in the ultrathin cross-sections of the common duct (CD) of D. citri. This study provides new information on D. citri mouthparts that may help to understand the behaviour of this important vector of HLB-associated bacteria. (C) 2011 Elsevier Ltd. All rights reserved.

Polymorphic toxin systems: Comprehensive characterization of trafficking modes, processing, mechanisms of action, immunity and ecology using comparative genomics

Background: Proteinaceous toxins are observed across all levels of inter-organismal and intra-genomic conflicts. These include recently discovered prokaryotic polymorphic toxin systems implicated in intra-specific conflicts. They are characterized by a remarkable diversity of C-terminal toxin domains generated by recombination with standalone toxin-coding cassettes. Prior analysis revealed a striking diversity of nuclease and deaminase domains among the toxin modules. We systematically investigated polymorphic toxin systems using comparative genomics, sequence and structure analysis. Results: Polymorphic toxin systems are distributed across all major bacterial lineages and are delivered by at least eight distinct secretory systems. In addition to type-II, these include type-V, VI, VII (ESX), and the poorly characterized "Photorhabdus virulence cassettes (PVC)", PrsW-dependent and MuF phage-capsid-like systems. We present evidence that trafficking of these toxins is often accompanied by autoproteolytic processing catalyzed by HINT, ZU5, PrsW, caspase-like, papain-like, and a novel metallopeptidase associated with the PVC system. We identified over 150 distinct toxin domains in these systems. These span an extraordinary catalytic spectrum to include 23 distinct clades of peptidases, numerous previously unrecognized versions of nucleases and deaminases, ADP-ribosyltransferases, ADP ribosyl cyclases, RelA/SpoT-like nucleotidyltransferases, glycosyltranferases and other enzymes predicted to modify lipids and carbohydrates, and a pore-forming toxin domain. Several of these toxin domains are shared with host-directed effectors of pathogenic bacteria. Over 90 families of immunity proteins might neutralize anywhere between a single to at least 27 distinct types of toxin domains. In some organisms multiple tandem immunity genes or immunity protein domains are organized into polyimmunity loci or polyimmunity proteins. Gene-neighborhood-analysis of polymorphic toxin systems predicts the presence of novel trafficking-related components, and also the organizational logic that allows toxin diversification through recombination. Domain architecture and protein-length analysis revealed that these toxins might be deployed as secreted factors, through directed injection, or via inter-cellular contact facilitated by filamentous structures formed by RHS/YD, filamentous hemagglutinin and other repeats. Phyletic pattern and life-style analysis indicate that polymorphic toxins and polyimmunity loci participate in cooperative behavior and facultative 'cheating' in several ecosystems such as the human oral cavity and soil. Multiple domains from these systems have also been repeatedly transferred to eukaryotes and their viruses, such as the nucleo-cytoplasmic large DNA viruses. Conclusions: Along with a comprehensive inventory of toxins and immunity proteins, we present several testable predictions regarding active sites and catalytic mechanisms of toxins, their processing and trafficking and their role in intra-specific and inter-specific interactions between bacteria. These systems provide insights regarding the emergence of key systems at different points in eukaryotic evolution, such as ADP ribosylation, interaction of myosin VI with cargo proteins, mediation of apoptosis, hyphal heteroincompatibility, hedgehog signaling, arthropod toxins, cell-cell interaction molecules like teneurins and different signaling messengers.

Imunocompetent Mice Model for Dengue Virus Infection

Dengue fever is a noncontagious infectious disease caused by dengue virus (DENV). DENV belongs to the family Flaviviridae, genus Flavivirus, and is classified into four antigenically distinct serotypes: DENV-1, DENV-2, DENV-3, and DENV-4. The number of nations and people affected has increased steadily and today is considered the most widely spread arbovirus (arthropod-borne viral disease) in the world. The absence of an appropriate animal model for studying the disease has hindered the understanding of dengue pathogenesis. In our study, we have found that immunocompetent C57BL/6 mice infected intraperitoneally with DENV-1 presented some signs of dengue disease such as thrombocytopenia, spleen hemorrhage, liver damage, and increase in production of IFN gamma and TNF alpha cytokines. Moreover, the animals became viremic and the virus was detected in several organs by real-time RT-PCR. Thus, this animal model could be used to study mechanism of dengue virus infection, to test antiviral drugs, as well as to evaluate candidate vaccines.

Pathogenicity of Metarhizium anisopliae towards Rhipicephalus (Boophilus) microplus under laboratory and field conditions

Metarhizium anisopliae is one of the most studied agents of biological control of several arthropod plagues, including the cattle tick Rhipicephalus (Boophilus) microplus. Studies have been conducted to assess the fungal complex infection process towards its hosts. To accomplish that, mutant strains overexpressing or lacking assumed determinant genes for the process were constructed over the years. A fundamental experiment to demonstrate a particular gene or set of genes participation is the bioassay. The comparison of bioassays using wild and engineered strains is an essential tool to affirm a given gene is crucial in the process. Therefore, the in vitro bioassays should mimic the results obtained in tests under field conditions. In this study, tests under laboratory and filed conditions were done and a correlation analysis was performed in order to statistically validate in vitro bioassays. Tick egg laying, larvae hatching and host mortality were recorded in each experiment through 21 days, both under laboratory and field conditions. In all cases, M. anisopliae treatments were statistically different from the control treatments. A linear regression analysis was performed between the cases. Laboratory results showed a statistically significant correlation with the field conditions using the Pearson's Correlation Test (P < 0.01 host mortality - 0.969, tick egg laying - 0.977 and larvae hatching - 0.956). These results legitimize the in vitro bioassays and, therefore, constitute them as a valid tool for studying this fungus behavior, so they can be used to infer M. anisopliae response towards R. (Boophilus) microplus.