Comparison of synganglion neuropeptides, neuropeptide receptors and neurotransmitter receptors and their gene expression in response to feeding in Ixodes scapularis (Ixodidae) vs. Ornithodoros turicata (Argasidae).

Illumina GAII high-throughput sequencing was used to compare expressed genes for female synganglion neuropeptides, neuropeptide receptors and neurotransmitter receptors of the soft tick Ornithodoros turicata with the hard tick Ixodes scapularis. Gene ontology molecular level three mapping revealed no significant differences amongst the same categories represented in O. turicata and I. scapularis. Transcripts predicting 22 neuropeptides or their receptors in the O. turicata synganglion were similar to annotations for 23 neuropeptides or receptors previously identified from I scapularis, with minor exceptions. A transcript predicting ecdysis triggering hormone receptor was identified in O. turicata; transcripts encoding for proprotein convertase and glycoprotein B were identified in both species. Transcripts predicting the same neurotransmitter receptors were found in the synganglion of both species. Gene expression of the transcripts showed numerous differences in response to feeding. Major differences were observed in expression of genes believed important in regulating slow vs. rapid feeding, blood water elimination, cuticle synthesis plasticity and in signalling reproductive activity. Although the glutamate receptor was strongly upregulated in both species, the gamma aminobutyric acid receptor, which inhibits glutamate, was upregulated significantly only in I. scapularis. These differences are consistent with the slow vs. rapid action of the pharyngeal pump in the two species.

Sustained efficacy of the novel topical repellent TT-4302 against mosquitoes and ticks.

Mosquitoes and ticks are blood-feeding pests of humans and animals that can vector many important aetiological agents of disease. Previous research demonstrated that TT-4302 (Guardian(®) Wilderness) containing 5% geraniol applied to human subjects gave 5-6 h of repellency against mosquitoes (depending on species) and was repellent to ticks in vitro. This study was conducted to obtain an independent third-party evaluation of TT-4302 against Stegomyia aegypti (= Aedes aegypti) (Diptera: Culicidae) mosquitoes and to test the efficacy of the product in the field against ticks. TT-4302 provided an average of 6.5 h of repellency of ≥ 95% [Weibull mean protection time: 7.4 h, 95% confidence interval (CI) 5.8-11.3 h] for St. aegypti, whereas a 15% DEET formulation provided 4.7 h of repellency (Weibull mean protection time: 5.2 h, 95% CI 3.7-6.9 h). In tick field trials, the efficacy of TT-4302 did not differ significantly from that of a 25% DEET formulation against Amblyomma americanum (Ixodida: Ixodidae). TT-4302 was 81.3% repellent at 2.5 h after application, whereas DEET was 77.2% repellent at the same time-point. Results at 3.5 h after application were 71.4% for TT-4302 and 72.9% for DEET.

Antiparasitic efficacy of a novel plant-based functional food using an Ascaris suum model in pigs.

Ascaris lumbricoides is the most prevalent soil-transmitted helminth (STH) infection of human beings worldwide. Chemotherapy with synthetic anthelmintics such as albendazole, mebendazole, and pyrantel pamoate is the current method of treatment; however, the emergence of anthelmintic resistance could substantially decrease the efficacy of such treatments and the sustainability of STH control programs. Additionally, benzimidazoles are not recommended for pregnant women or children under age one. A blinded, controlled study was conducted to evaluate the efficacy of two microencapsulated, plant-based essential oil blends, TTN1013 (α-pinene, linalyl acetate, p-cymene, and thymol octanoate) and TTN1014 (α-pinene, linalyl acetate, p-cymene, and thymol acetate) as functional foods against Ascaris suum infection in pigs, an important pathogen that closely resembles human infections with A. lumbricoides. Four groups of 16 female, 21-24 day old, Yorkshire-cross pigs were treated daily with 0.5 or 1.0mg/kg TTN1013, 1.0mg/kg TTN1014, or 1.0mg/kg equivalent of empty capsules, delivered inside a cream-filled sandwich cookie for 14 weeks. Three days after the initiation of daily treatments, pigs were inoculated daily with A. suum eggs for four weeks. Pigs were weighed weekly and fecal egg counts (FEC) were conducted weekly starting five weeks after initial inoculation with A. suum eggs. Fourteen weeks after first infection with eggs, pigs were necropsied and worms were recovered, counted and separated according to sex. TTN1013 administered daily at a dose of 1.0mg/kg yielded a statistically significant reduction in total worm counts (76.8%), female worm counts (75.5%), FEC (68.6%), and worm volume (62.9%) when compared to control group. Reduction of total and female worm numbers and FEC were not significant for TTN1014 or at the 0.5mg/kg dose of TTN1013. All treatments were well-tolerated by all pigs and did not cause any adverse reactions. All pigs remained clinically normal and showed no signs of reduced intestinal health for the duration of treatment. Based on these results, TTN1013 shows promise as a daily supplement to reduce infection burdens of soil transmitted helminths in both pigs and human beings.

Performance of the plant-based repellent TT-4302 against mosquitoes in the laboratory and field and comparative efficacy to 16 mosquito repellents against Aedes aegypti (Diptera: Culicidae).

Repellent efficacy of the plant-based repellent, TT-4302 (5% geraniol), was compared with 16 other products in laboratory arm-in-cage trials against Aedes aegypti (L). Eight repellents (Badger, BioUD, Burt's bees, California Baby, Cutter Natural, EcoSMART, Herbal Armor, and SkinSmart) exhibited a mean repellency below 90% to Ae. aegypti at 0.5 h after application. Three repellents (Buzz Away Extreme, Cutter Advanced, and OFF! Botanicals lotion) fell below 90% repellency 1.5 h after application. TT-4302 exhibited 94.7% repellency 5 h posttreatment, which was a longer duration than any of the other repellents tested. The positive control, 15% DEET (OFF! Active), was repellent for 3 h before activity dropped below 90%. Additional arm-in-cage trials comparing TT-4302 with 15% DEET were carried out against Anopheles quadrimaculatus Say. At 6 h after treatment, TT-4302 provided 95.2% repellency while DEET exhibited 72.2%. In North Carolina field trials, TT-4302 provided 100% repellency 5 h after application against Aedes albopictus Skuse while DEET provided 77.6% repellency. These results demonstrate that TT-4302 is an efficacious plant-based repellent that provides an extended duration of protection compared with many other commercially available products.

Activity of the plant-based repellent, TT-4302 against the ticks Amblyomma americanum, Dermacentor variabilis, Ixodes scapularis and Rhipicephalus sanguineus (Acari: Ixodidae).

The plant-based repellent TT-4302 (5 % geraniol) was compared to deet (15 %) in laboratory two-choice bioassays against the ticks Amblyomma americanum, Dermacentor variabilis, Ixodes scapularis, and Rhipicephalus sanguineus. At 2.5 and 3.5 h after treatment of filter paper with TT-4302, 100 % repellency was observed for all species at both time points with the exception of I. scapularis at the 3.5 h evaluation where repellency was 95.8 %. Deet was 100 % repellent at both time points for D. variabilis and R. sanguineus and was 100 % repellent at the 2.5 h evaluation for I. scapularis. Repellency of deet to A. americanum was 88.9 and 95.8 % at 2.5 and 3.5 h, respectively which was not significantly different than that of TT-4302. No significant difference against I. scapularis was observed between TT-4302 and deet at 3.5 h after treatment where deet was 87.5 % repellent. A variant of TT-4302, TT-4228 was tested in the laboratory against A. americanum and was compared to deet (15 %) in field trials against wild populations of ticks in North Carolina, USA. In the laboratory, TT-4228 was 94.4 and 87.5 % repellent at 2.5 and 3.5 h after treatment, respectively. In the field where the predominant tick species was A. americanum, significantly fewer ticks were collected from socks worn by human volunteers that were treated with TT-4228 compared to those treated with deet 2.5 or 3.5 h after treatment. Significantly fewer ticks were recovered from socks treated with TT-4228 than their paired untreated controls 2.5 or 3.5 h after treatment and repellencies were 90 and 70 %, respectively. Fewer ticks were collected from deet-treated compared to their paired untreated socks 2.5 h after application; however, no significant difference was found in the number of ticks collected from deet-and untreated socks 3.5 h after treatment.

Synganglion transcriptome and developmental global gene expression in adult females of the American dog tick, Dermacentor variabilis (Acari: Ixodidae).

454 Pyrosequencing was used to characterize the expressed genes from the synganglion and associated neurosecretory organs of unfed and partially fed virgin and mated replete females of the American dog tick, Dermacentor variabilis. A total of 14,881 contiguous sequences (contigs) was assembled, with an average size of 229 bp. Gene ontology terms for Level 2 biological processes were assigned to 4366 contigs. Seven acetylcholinesterases, a muscarinic acetylcholine (ACh) receptor, two nicotinic ACh receptor ß-subunits, two ACh unc-18 regulators, two dopamine receptors, two gamma aminobutyric acid (GABA) receptors, two GABA transporters, two norepinephrine transporters and an octopamine receptor are described. Microarrays were conducted to examine global gene expression and quantitative real-time polymerase chain reaction was used to verify expression of selected neuropeptides. Hierarchical clustering of all differentially expressed transcripts grouped part-fed and replete ticks as being more similar in terms of differentially expressed genes with unfed ticks as the outgroup. Nine putative neuropeptides (allatostatin, bursicon-ß, preprocorazonin, glycoprotein hormone α, insulin-like peptide, three orcokinins, preprosulphakinin) and a gonadotropin releasing hormone receptor were differentially expressed, and their developmental expression and role in reproduction was investigated. The presence of eclosion hormone, corazonin and bursicon in the synganglion, which in insects regulate behaviour and cuticle development associated with moulting, suggest that this system may be used in ticks to regulate blood feeding, cuticle expansion and development related to female reproduction; adult ticks do not moult.

First report of the repellency of 2-tridecanone against ticks.

2-Tridecanone and 2-undecanone are both found naturally in the trichomes of wild tomato plants and are important in plant resistance to herbivory. 2-Undecanone is the U.S. Environmental Protection Agency (EPA)-registered active ingredient in the commercially available arthropod repellent, BioUD(®) . The goal of this study was to examine the tick repellency of 2-tridecanone. Two-choice bioassays were conducted using 8% 2-tridecanone vs. the repellent carrier (absolute ethanol) and compared with two-choice studies using 8% 2-undecanone vs. absolute ethanol. Unfed, host-seeking adult (mixed sex) Amblyomma americanum (L.) (Acari: Ixodidae) and Dermacentor variabilis Say (Acari: Ixodidae) were used to evaluate repellency and time to repellent failure at room temperature. The present study shows in filter paper assays (0.63 mg test compound/cm(2) ) that 2-tridecanone was 87% repellent to A. americanum at 12 h after application, but had no statistically significant repellency at 15 h and 24 h, and was 72% repellent to D. variabilis at 15 h, but had no statistically significant repellency at 24 h. By contrast, 2-undecanone was 74% and 75% repellent to A. americanum and D. variabilis, respectively, at 2 h after application, but no statistically significant repellency was noted at 2.5 h and 3 h. In two-choice assays on cheesecloth, 2-tridecanone at 0.25 mg/cm(2) was 85% repellent to A. americanum 6 h after application, demonstrating its potential use as an arthropod repellent that can be used on clothing without the need for formulation. No statistically significant repellency was found at 9 h or 12 h. The potential use of 2-tridecanone as a tick repellent is discussed.

Novel field assays and the comparative repellency of BioUD(®) , DEET and permethrin against Amblyomma americanum.

Two new field bioassay methods were developed to compare the repellent activity of BioUD(®) (containing 7.75% 2-undecanone), 98.1% DEET and 0.5% permethrin against natural populations of nymphal Amblyomma americanum (L.) (Acari: Ixodidae). In a cloth sheet assay, pieces of material measuring 41 × 58 cm, separately treated with one of the test materials or the appropriate solvent carrier, were placed at random on the ground and baited with dry ice for 1 h. Mean numbers of ticks on repellent-treated sheets were significantly lower than on control sheets. There was no significant difference in the number of ticks collected between sheets treated with BioUD(®) and those treated with DEET. However, significantly fewer ticks were found on sheets treated with BioUD(®) or DEET than on permethrin-treated sheets. In a sock test, over-the-calf tube socks were treated with one of the test materials or the appropriate solvent carrier. Human volunteers wore a repellent-treated and a corresponding carrier-treated sock on either leg and walked randomly over an area of approximately 4000 m(2) for 15 min. Significantly fewer ticks were collected from socks treated with BioUD(®) or DEET than from socks treated with the carrier and there was no significant difference in repellency between these two agents. No difference in the mean number of ticks collected was found between permethrin-treated and corresponding carrier-treated socks. To examine the mechanism of repellency of BioUD(®) , a four-choice olfactometer was used to assess spatial repellency against adult A. americanum. As expected in the absence of a repellent, when all choices were represented by water-treated filter paper, ticks were equally distributed among the choices. When one choice consisted of BioUD(®) -treated filter paper and the remaining choices of water-treated paper, the distribution of ticks on the repellent-treated paper was significantly lower than might be expected to occur by chance, suggesting that repellency is at least partly achieved by an olfactory mechanism.

Efficacy of the new repellent BioUD against three species of ixodid ticks.

BioUD with the active ingredient 2-undecanone originally derived from wild tomato plants is a new repellent recently registered by the US EPA. Repellent efficacy of BioUD (7.75% 2-undecanone) and DEET (98.11%) was examined in the laboratory using a choice test between repellent-treated and control filter paper surfaces for Amblyomma americanum, Dermacentor variabilis, and Ixodes scapularis. BioUD provided greater repellency against A. americanum and I. scapularis than DEET. No difference was found between BioUD and DEET against D. variabilis. In head-to-head assays between BioUD and DEET, undiluted and 50% dilutions of BioUD were more repellent than undiluted DEET against all three species tested. Similarly, a 25% dilution of BioUD was more repellent than DEET against A. americanum while no difference in mean percentage repellency was found between a 25% dilution of BioUD and DEET against I. scapularis. Based on regression analysis, the concentration of BioUD required for equivalent repellency to 98.11% DEET was 39.5% for D. variabilis and 29.7% for I. scapularis. A log-probit model could not be constructed for A. americanum from the dosages tested. Based on filter paper head-to-head assays, BioUD is at least 2-4 times more active as a repellent than DEET against three species of ixodid ticks under the conditions of our laboratory bioassays.