Role of Calcium, Catecholamines and Cyclic Amp in Controlling Salivary Fluid Secretion in the Lone Star Tick

Entomolog

Cold-hardiness of a laboratory colony of lone star ticks.

The cold-hardiness of a lone star tick, Ambylomma americium (L.) laboratory colony was characterized. Fed and unfed larvae, fed and unfed nymphs, and unfed adults did not survive exposure to -17C for 7 d. After an 8-d exposure to -10C, adults tolerated cold better than immatures and unfed specimens fared better than fed ticks. Exposing unfed 6-wk old (post mold) adult males and females to -15C for increasing intervals up to 2 hrs suggests that males were more tolerant to cold then were females. Half of all adults were alive 3 d after the 2-h low-temperature treatment. Males may have survived because of a significantly higher hemolymph osmotic pressure, although the solute concentration increased for both sexes after a 2-h exposure to 0C. Acclimation to 5C for 7 d had no influence on supercooling points for unfed males and females, engorged nymphs and larvae, and eggs. None of the life stages survived supercooling, which strongly suggests that this species is freeze-intolerant. Intolerance of immature stages to chilling may be a limiting factor in the northern distribution of lone star ticks in North America

Emergence of Ixodes scapularis and Borrelia burgdorferi, the Lyme disease vector and agent, in Ohio

Lyme disease, the most common vector-borne disease in the United States, is caused by a tick-borne infection with Borrelia burgdorferi. Currently, Ohio is considered by the Centers for Disease Control and Prevention (CDC) to be non-endemic for Lyme disease. The low incidence of Lyme disease in this state was largely attributed to the absence of the transmitting vector, Ixodes scapularis, commonly known as the blacklegged tick. However, a tick surveillance program established by Ohio Department of Health indicated that the number of I. scapularis ticks in Ohio had increased sharply in recent years, from 0 - 5 ticks per year during 1983 - 2008 to 15 in 2009, 40 in 2010, and 184 in 2011. During the fall deer hunting season, examination of deer heads submitted to Ohio Department of Agriculture found 29 I. scapularis from 7 counties in 2010 and 1,830 from 25 counties in 2011. As of 2012, the tick had been found in 57 of the 88 counties of Ohio. In addition, all three active stages (larva, nymph, and adult) of I. scapularis were found in Tiverton Township of Coshocton County, demonstrating the presence of established tick populations at this central Ohio location. Of 530 nymphal or adult I. scapularis analyzed by quantitative polymerase chain reaction (qPCR), 32 (6.1%) tested positive for the B. burgdorferi flaB gene, ranging from 36 to 390,000 copies per tick. Antibodies to B. burgdorferi antigens were detected in 2 of 10 (20%) field-captured Peromyscus leucopus from Tiverton Township, and in 41 of 355 (11.5%) dogs residing in Ohio. Collectively, these data suggest that the enzootic life cycle of B. burgdorferi has become established in Ohio, which poses risk of Lyme disease to people and animals in the area

Quantitative Factors Proposed to Influence the Prevalence of Canine Tick-Borne Disease Agents in the United States

The Companion Animal Parasite Council hosted a meeting to identify quantifiable factors that can influence the prevalence of tick-borne disease agents among dogs in North America. This report summarizes the approach used and the factors identified for further analysis with mathematical models of canine exposure to tick-borne pathogens

Para-infectious brain injury in COVID-19 persists at follow-up despite attenuated cytokine and autoantibody responses

To understand neurological complications of COVID-19 better both acutely and for recovery, we measured markers of brain injury, inflammatory mediators, and autoantibodies in 203 hospitalised participants; 111 with acute sera (1–11 days post-admission) and 92 convalescent sera (56 with COVID-19-associated neurological diagnoses). Here we show that compared to 60 uninfected controls, tTau, GFAP, NfL, and UCH-L1 are increased with COVID-19 infection at acute timepoints and NfL and GFAP are significantly higher in participants with neurological complications. Inflammatory mediators (IL-6, IL-12p40, HGF, M-CSF, CCL2, and IL-1RA) are associated with both altered consciousness and markers of brain injury. Autoantibodies are more common in COVID-19 than controls and some (including against MYL7, UCH-L1, and GRIN3B) are more frequent with altered consciousness. Additionally, convalescent participants with neurological complications show elevated GFAP and NfL, unrelated to attenuated systemic inflammatory mediators and to autoantibody responses. Overall, neurological complications of COVID-19 are associated with evidence of neuroglial injury in both acute and late disease and these correlate with dysregulated innate and adaptive immune responses acutely

Para-infectious brain injury in COVID-19 persists at follow-up despite attenuated cytokine and autoantibody responses

To understand neurological complications of COVID-19 better both acutely and for recovery, we measured markers of brain injury, inflammatory mediators, and autoantibodies in 203 hospitalised participants; 111 with acute sera (1–11 days post-admission) and 92 convalescent sera (56 with COVID-19-associated neurological diagnoses). Here we show that compared to 60 uninfected controls, tTau, GFAP, NfL, and UCH-L1 are increased with COVID-19 infection at acute timepoints and NfL and GFAP are significantly higher in participants with neurological complications. Inflammatory mediators (IL-6, IL-12p40, HGF, M-CSF, CCL2, and IL-1RA) are associated with both altered consciousness and markers of brain injury. Autoantibodies are more common in COVID-19 than controls and some (including against MYL7, UCH-L1, and GRIN3B) are more frequent with altered consciousness. Additionally, convalescent participants with neurological complications show elevated GFAP and NfL, unrelated to attenuated systemic inflammatory mediators and to autoantibody responses. Overall, neurological complications of COVID-19 are associated with evidence of neuroglial injury in both acute and late disease and these correlate with dysregulated innate and adaptive immune responses acutely

Fluid secretion by isolated tick salivary glands dependent on an intact cytoskeleton

Isolated salivary glands from female Dermacentor variabilis (Say) were pre-treated with either cytochalasin D or nocodazol, followed by stimulation with dopamine. Glands pre-treated with 1 μM-cytochalasin D did not secrete fluid; pre-treatment with 1 nM-cytochalasin D did not significantly reduce fluid secretion. Glands pre-treated with 1 μM-nocodazol had a significant reduction in fluid secretion (P = 0.003); 1 nM-nocodazol did not significantly affect fluid secretion. Ligation of the main salivary duct and pre-treatment with 1 μm-cytochalasin D significantly increased gland weight compared to the dopamine stimulated controls (P = 0.0412). Cytochalasin D-treated type III acini had a significantly larger mean diameter compared to the dopamine control (P = 0.0047). Glands treated with 1 mm-verapamil plus 10 μm-dopamine exhibited a significant decrease in fluid secretion (P = 0.017), and when ligated, had a significantly decreased weight compared to the controls (P = 0.0028). © 1994

Evidence that dilation of isolated salivary ducts from the tick Dermacentor variabilis (Say) is mediated by nitric oxide

We used pharmacological methods to test the hypothesis that female Dermacentor variabilis salivary ducts dilated when dopamine-stimulated and that dilation was nitric oxide-mediated. Stimulation with dopamine resulted in an increased diameter (19.7%) compared to unstimulated ducts (P\u3c0.005). Pretreatment with L-NAME, an inhibitor of nitric oxide synthase, or cytochalasin D abolished the dilation. Addition of L-arginine to L-NAME- treated ducts partially restored the ability to dilate. A cuticular coil composed of a series of concentric rings ran the length of the duct adjacent to the epithelial cell layer. In stimulated ducts, the center-to-center periodicity of these rings increased from 0.59 μm in unfed ducts to 1.0 μm from partially fed ducts (P\u3c0.05). When the ducts from partially fed females were stimulated with dopamine, the periodicity increased further to 1.75 μm (P\u3c0.05), suggesting the coils moved further apart in response to stimulation. Prominent folds lining the lumen of unstimulated ducts were less pronounced in stimulated preparations, suggesting that the cuticle stretches, thereby increasing lumen size. Actin was localized in epithelial cells as a honeycomb pattern that we suggest links the epithelial cells to the rings. Together, these data support the following hypothesis: stimulated ducts dilated during fluid production; dilation involved an actin-based system, and was mediated by nitric oxide. Dilation of the duct may enhance its role as a reservoir for saliva produced by the acini during the period between imbibition and salivation. (C) 2000 Elsevier Science Ltd

Tick Acquisition of Ehrlichia canis from Dogs Treated with Doxycycline Hyclate▿

Doxycycline generally alleviates clinical monocytic ehrlichiosis, but its efficacy in the control of monocytotropic ehrlichial pathogens requires further investigation. In this study, Ehrlichia canis was detected in dogs treated with doxycycline for 14 days and in ticks fed on these dogs, suggesting that treated dogs can remain reservoirs for E. canis

Detection of Ehrlichia canis in Canine Carrier Blood and in Individual Experimentally Infected Ticks with a p30-Based PCR Assay

Detection of vector-borne pathogens is necessary for investigation of their association with vertebrate and invertebrate hosts. The ability to detect Ehrlichia spp. within individual experimentally infected ticks would be valuable for studies to evaluate the relative competence of different vector species and transmission scenarios. The purpose of this study was to develop a sensitive PCR assay based on oligonucleotide sequences from the unique Ehrlichia canis gene, p30, to facilitate studies that require monitoring this pathogen in canine and tick hosts during experimental transmission. Homologous sequences for Ehrlichia chaffeensis p28 were compared to sequences of primers derived from a sequence conserved among E. canis isolates. Criteria for primer selection included annealing scores, identity of the primers to homologous E. chaffeensis sequences, and the availability of similarly optimal primers that were nested within the target template sequence. The p30-based assay was at least 100-fold more sensitive than a previously reported nested 16S ribosomal DNA (rDNA)-based assay and did not amplify the 200-bp target amplicon from E. chaffeensis, the human granulocytic ehrlichiosis agent, or Ehrlichia muris DNA. The assay was used to detect E. canis in canine carrier blood and in experimentally infected Rhipicephalus sanguineus ticks. Optimized procedures for preparing tissues from these hosts for PCR assay are described. Our results indicated that this p30-based PCR assay will be useful for experimental investigations, that it has potential as a routine test, and that this approach to PCR assay design may be applicable to other pathogens that occur at low levels in affected hosts