Amblyomma maculatum-associated rickettsiae in vector tissues and vertebrate hosts during tick feeding.

Rickettsia parkeri, a causative agent of spotted fever rickettsiosis, is transmitted by Amblyomma maculatum (Gulf Coast tick), a tick that may also carry a non-pathogenic spotted fever group Rickettsia, "Candidatus Rickettsia andeanae". Here, we evaluated R. parkeri and "Candidatus R. andeanae" in tissues from A. maculatum prior to, during, and after blood feeding on rabbits. Using colony-reared A. maculatum that were capillary-fed uninfected cells, R. parkeri, "Candidatus R. andeanae", or both rickettsiae, we detected higher levels of Rickettsia spp. in the respective treatment groups. Rickettsial levels increased during blood feeding for both R. parkeri and "Candidatus R. andeanae", with a greater increase in R. parkeri in co-infected ticks compared to singly-infected ticks. We detected transovarial transmission of "Candidatus R. andeanae" in egg and larval cohorts and confirmed vertical transmission of R. parkeri in one group of larvae. Rabbits from all Rickettsia-exposed groups seroconverted on immunofluorescent antibody testing using R. parkeri antigen. Visualization of "Candidatus R. andeanae" in tick salivary glands suggested potential transmission via tick feeding. Here, rickettsial levels in artificially infected ticks demonstrate changes during feeding and transovarial transmission that may be relevant for interpreting rickettsial levels detected in wild A. maculatum.

Apoplast proteome reveals that extracellular matrix contributes to multistress response in poplar.

BACKGROUND: Riverine ecosystems, highly sensitive to climate change and human activities, are characterized by rapid environmental change to fluctuating water levels and siltation, causing stress on their biological components. We have little understanding of mechanisms by which riverine plant species have developed adaptive strategies to cope with stress in dynamic environments while maintaining growth and development. RESULTS: We report that poplar (Populus spp.) has evolved a systems level "stress proteome" in the leaf-stem-root apoplast continuum to counter biotic and abiotic factors. To obtain apoplast proteins from P. deltoides, we developed pressure-chamber and water-displacement methods for leaves and stems, respectively. Analyses of 303 proteins and corresponding transcripts coupled with controlled experiments and bioinformatics demonstrate that poplar depends on constitutive and inducible factors to deal with water, pathogen, and oxidative stress. However, each apoplast possessed a unique set of proteins, indicating that response to stress is partly compartmentalized. Apoplast proteins that are involved in glycolysis, fermentation, and catabolism of sucrose and starch appear to enable poplar to grow normally under water stress. Pathogenesis-related proteins mediating water and pathogen stress in apoplast were particularly abundant and effective in suppressing growth of the most prevalent poplar pathogen Melampsora. Unexpectedly, we found diverse peroxidases that appear to be involved in stress-induced cell wall modification in apoplast, particularly during the growing season. Poplar developed a robust antioxidative system to buffer oxidation in stem apoplast. CONCLUSION: These findings suggest that multistress response in the apoplast constitutes an important adaptive trait for poplar to inhabit dynamic environments and is also a potential mechanism in other riverine plant species.

Retention of gadolinium in the brains of healthy dogs after a single intravenous administration of gadodiamide.

OBJECTIVE To determine brain region affinity for and retention of gadolinium in dogs after administration of gadodiamide and whether formalin fixation affects quantification. ANIMALS 14 healthy dogs. PROCEDURES 13 dogs received gadodiamide (range, 0.006 to 0.1 mmol/kg, IV); 1 control dog received a placebo. Dogs received gadodiamide 3 to 7 days (n = 8) or 9 hours (5) before euthanasia and sample collection. Brain regions were analyzed with inductively coupled mass spectrometry (ICP-MS) and transmission electron microscopy. Associations between dose, time to euthanasia, and gadolinium retention quantities (before and after fixation in 5 dogs) were evaluated. RESULTS Gadolinium retention was seen in all brain regions at all doses, except for the control dog. Exposure 3 to 7 days before euthanasia resulted in 1.7 to 162.5 ng of gadolinium/g of brain tissue (dose-dependent effect), with cerebellum, parietal lobe, and brainstem affinity. Exposure 9 hours before euthanasia resulted in 67.3 to 1,216.4 ng of gadolinium/g of brain tissue without dose dependency. Transmission electron microscopy revealed gadolinium in examined tissues. Fixation did not affect quantification in samples immersed for up to 69 days. CONCLUSIONS AND CLINICAL RELEVANCE Gadodiamide exposure resulted in gadolinium retention in the brain of healthy dogs. Cerebellum, parietal lobe, and brainstem affinity was detected with dose dependency only in dogs exposed 3 to 7 days before euthanasia. Fixation had no effect on quantification when tissues were immersed for up to 69 days. Physiologic mechanisms for gadolinium retention remained unclear. The importance of gadolinium retention requires further investigation.

Transmission dynamics of Borrelia turicatae from the arthropod vector.

BACKGROUND: With the global distribution, morbidity, and mortality associated with tick and louse-borne relapsing fever spirochetes, it is important to understand the dynamics of vector colonization by the bacteria and transmission to the host. Tick-borne relapsing fever spirochetes are blood-borne pathogens transmitted through the saliva of soft ticks, yet little is known about the transmission capability of these pathogens during the relatively short bloodmeal. This study was therefore initiated to understand the transmission dynamics of the relapsing fever spirochete Borrelia turicatae from the vector Ornithodoros turicata, and the subsequent dissemination of the bacteria upon entry into murine blood. METHODOLOGY/PRINCIPAL FINDINGS: To determine the minimum number of ticks required to transmit spirochetes, one to three infected O. turicata were allowed to feed to repletion on individual mice. Murine infection and dissemination of the spirochetes was evaluated by dark field microscopy of blood, quantitative PCR, and immunoblotting against B. turicatae protein lysates and a recombinant antigen, the Borrelia immunogenic protein A. Transmission frequencies were also determined by interrupting the bloodmeal 15 seconds after tick attachment. Scanning electron microscopy (SEM) was performed on infected salivary glands to detect spirochetes within acini lumen and excretory ducts. Furthermore, spirochete colonization and dissemination from the bite site was investigated by feeding infected O. turicata on the ears of mice, removing the attachment site after engorment, and evaluating murine infection. CONCLUSION/SIGNIFICANCE: Our findings demonstrated that three ticks provided a sufficient infectious dose to infect nearly all animals, and B. turicatae was transmitted within seconds of tick attachment. Spirochetes were also detected in acini lumen of salivary glands by SEM. Upon host entry, B. turicatae did not require colonization of the bite site to establish murine infection. These results suggest that once B. turicatae colonizes the salivary glands the spirochetes are preadapted for rapid entry into the mammal.