Chemical profile, characterization and acaricidal activity of essential oils of three plant species and their nanoemulsions against Tyrophagus putrescentiae , a stored-food mite Abstract Essential oils of Ocimum basilicum (L.), Achillea fragrantissima (Forssk.) and Achillea santolina (L.) were obtained by hydrodistillation and analyzed using gas chromatography (GC) and GC/mass spectrometry (MS). Oil-in-water nanoemulsions (10% active ingredient) were prepared through a high-energy (ultrasonication) emulsification process. Nanoemulsions were characterized by viscosity, pH, thermodynamic stability, droplet size, polydispersity index (PDI) and scanning electron microscopy (SEM) measurements. The plant oils and their nanoemulsions showed considerable acaricidal activity against the mold mite, Tyrophagus putrescentiae (Schrank) (Sarcoptiformes: Acaridae). In a contact toxicity bioassay and 48 h post treatment, O. basilicum oil was the most toxic, followed by A. fragrantissima and A. santolina, where LC50 values were 8.4, 14.1 and 21.8 µl/cm2, respectively. LC50 for benzyl benzoate, a standard acaricide was 9.8 µl/cm2. Upon fumigation, responses also varied according to the test oil. Based on the 48-h LC50 values, the same manner of activity was also observed, where O. basilicum was the most toxic followed by A. fragrantissima and A. santolina. When prepared as nanoemulsions (particle size from 78.5 to 104.6) and tested as fumigants, toxicity of the oils was increased drastically with LC50 values of 2.2, 4.7, and 9.6 µl/l air for O. basilicum, A. fragrantissima and A. santolina, respectively. The oils showed a moderate to strong residual acaricidal activity, where O. basilicum oil was the most effective. The results suggest that appropriate nanoemulsions containing the tested oils can be developed to control T. putrescentiae after the required toxicological assessments.

Tick infestation on caimans: a casual tick-host association in the Atlantic rainforest biome? Abstract Ticks are parasites of birds, mammals, amphibians, and reptiles, but information about tick communities that parasitize reptiles in the Neotropical region is still fragmentary. In the present study, we assessed the presence of ticks on broad-snouted caimans (Caiman latirostris) and Cuvier’s dwarf caimans (Paleosuchus palpebrosus) trapped in the Atlantic rainforest biome in Pernambuco state, north-eastern Brazil, to determine which tick species feed on these animals and how frequent or rare this parasite-wildlife association is. We also report an occasional finding of Amblyomma rotundatum on a smooth-fronted caiman (Paleosuchus trigonatus) in the Amazon biome in Pará state, northern Brazil. Out of 490 animals trapped in the Atlantic rainforest biome, four (0.82%) broad-snouted caimans were infested by ticks. Ticks belonged to two Amblyomma species: A. rotundatum (three females) and A. fuscum (one male). Our findings indicate that ticks are infrequent parasites of caimans in the Atlantic rainforest biome. Tick infestation on broad-snouted caimans is probably of minor clinical significance and probably a casual finding due to habitat sharing with the common tick hosts.

Response of Suidasia pontifica (Acaridida: Suidasiidae) to phosphine fumigation Abstract The present study evaluated the level of phosphine toxicity of various strains of Suidasia pontifica, infesting stored agricultural products in the Philippines. The bioassays for the efficacy of phosphine fumigation followed the FAO standard method. A full assay comprised eight concentrations ranging from 0.005 to 1.00 mg/L with 20, 72 and 144-h exposure periods. A total of 15 strains were collected, representing 15 provinces in the country. Results revealed that the most susceptible strain (MR12gsn) was recorded from a private owned warehouse in General Santos City—LC50 and LC99 were 0.009 and 0.025 mg/L, respectively. On the other hand, the most resistant strain (LR3tar) was collected from a private owned warehouse in Tarlac City—LC50 and LC99 were 1.501 and 2.407 mg/L, based on discrimination dose tests. Eggs were more tolerant than the adults; complete mortality of eggs was achieved at 0.50 mg/L for 72 h and 0.35 mg/L for 144 h, whereas complete mortality of adults was obtained with 0.10 and 0.20 mg/L for 144 and 72 h exposure, respectively. The estimated minimum effective concentration (MEC) of phosphine to give complete kill of S. pontifica at various exposure periods must exceed 0.50 mg/L for 72 and 144 h. These results simply illustrate the increased efficiency (i.e., increased mortality) of phosphine fumigation of longer exposure of S. pontifica to lower concentrations.

A standardized method for the construction of a tick drag/flag sampling approach and evaluation of sampling efficacy Abstract Drag sampling and flagging are two of the most effective and widely applied techniques to monitor tick populations. Despite the importance of this sampling strategy, there is a lack of standardized protocols for the construction of an inexpensive tick drag/flag. To this end, we provide a step-by-step protocol that details the construction of a tick drag/flag. We provide evidence of efficacy by comparing results obtained over 3-months at 108 locations within the William B. Bankhead National Forest, Alabama, USA. Overall, our drag/flag sampling approach yielded 1127 larvae, 460 nymphs, and 53 adults for a total of 1640 ticks representing three species. We detected significant patterns in Amblyomma americanum abundance for nymphs and adults with greater counts in June (β = 0.91 ± 0.36, 95% CI 0.55–1.27; β = 2.44 ± 0.63, 95% CI 1.81–3.07, respectively) and July (β = 0.73 ± 0.36, 95% CI 0.37–1.09; β = 1.65 ± 0.66, 95% CI 0.99–2.31, respectively) as compared to August. We also detected a significant difference in tick captures by tick drag/flag fabric type with greater captures when muslin was used as compared to flannel (β = 1.07 ± 0.06, 95% CI 1.01–1.13). Our goal is to provide instructions to assemble a highly effective tick drag/flag using minimal supplies. Evaluation and improvements of sampling techniques is essential to understand impacts of landscape management and larger stressors, such as climate change on tick populations but also for enhancing detection of invasive non-native species.

Life history of Macrocheles muscaedomesticae (Parasitiformes: Macrochelidae): new insights on life history and evidence of facultative parasitism on Drosophila Abstract Macrocheles muscaedomesticae is a cosmopolitan macrochelid mite whose populations have likely diverged considering the many locations they inhabit, but most of the work published on this mite species has been on the basis of their association with the house fly, Musca domestica. Here, we studied several aspects of the biology of M. muscaedomesticae associated with drosophilid flies collected in Alberta, Canada. We assessed the degree of divergence of our populations from others, compared their life history to other published populations and experimentally tested whether M. muscaedomesticae feeds on Drosophila hydei hosts by comparing the body mass of mites that attached to hosts to those that did not. There was no strong phylogenetic differentiation among any of the M. muscaedomesticae specimens, suggesting multiple recent introductions of this species to Canada. Compared to other populations, our mites exhibited lower fecundity, which may have been a result of the temperature or nematode-only diet in which they were maintained. Finally, mites that attached to hosts for 4 h weighed significantly more than those that did not. Without direct evidence for host tissue transfer to the mites, it is difficult to determine whether the mites are indeed feeding on their hosts while attached. However, the existing evidence for the costs fly hosts endure at the expense of these mites makes this relationship antagonistic.

Choice test to determine potential attractants and repellents for the sheep scab mite, Psoroptes ovis (Acari: Psoroptidae) Abstract A choice test bioassay was devised to screen compounds as potential semiochemicals (e.g., kairomones or allomones that mediate aggregation, attraction or repellence) for the obligate parasitic mite, Psoroptes ovis. The choice test used filter paper discs in a 1:4 test:control ratio and was found to be a reliable, effective and efficient method. Four mammalian lipid components were assessed as potential attractants—linoleic acid, arachidonic acid, methyl myristate and squalene—, and the insect/tick repellent DEET for potential repellence. Linoleic acid was significantly attractive to P. ovis adult females and has the potential to act as an attractant. Identification of P. ovis semiochemicals, especially attractants, would be beneficial in the development of novel control methods and tools for this species. This is essential considering the increase in resistance to the limited prophylactic chemical treatments in the UK, and the high prevalence of scab infections.

Genetic diversity of Haemaphysalis longicornis from China and molecular detection of Rickettsia Abstract The tick Haemaphysalis longicornis (Neumann) (Acari, Ixodidae) is distributed throughout China and is the most notorious blood feeding ectoparasite of livestock. Haemaphysalis longicornis can transmit a large number of pathogens that cause human infectious diseases, such as Rickettsia spp. Here, we characterized the genetic structure of H. longicornis and tested for the presence of Rickettsia spp. from five regions in China. Analysis of the two mitochondrial marker sequences (16S rRNA and COI) and the nuclear sequence (ITS2) showed that the overall level of nucleotide diversity was low and the variability did not differ among the five regions. From the five locations, the infection rates of Rickettsia species ranged from 0 to 65%. The nucleotide diversities of the high-infected group were lower than those of the low- and uninfected group. And in neutrality tests for the high-infected group based on COI sequences, the Tajima’s D and Fu’s FS were coincidentally < 0 and significant, whereas they were closer to zero and non-significant in low- and uninfected groups.

Expression dynamics of autophagy-related genes in the cattle tick Rhipicephalus microplus during embryonic development and under increasing larval starvation Abstract Rhipicephalus microplus is a hematophagous ectoparasite that significantly affects parasitized cattle. As a one-host tick its entire life cycle consists of free-living and parasitic forms. Its extraordinary ability to survive during prolonged off-host periods has been related to the process of cytoplasmic degradation called autophagy. In order to deepen our understanding of this process during R. microplus non-parasitic stages, we determined the expression dynamics of a set of five autophagy-related genes (ATG genes) during embryonic development and over an increasing larval starvation period of 50 days. We found two apparent successive waves of ATG genes transcriptional activation, which paralleled key embryonic changes such as cellularization and organogenesis, as well as nutrient utilization. Moreover, during increasing larval starvation, ATG genes were up-regulated cyclically every 10–15 days. Taken together, our results suggest that autophagy is playing a major role in embryo development and energy metabolism during starvation in R. microplus.

Behavioral responses of Ixodes scapularis tick to natural products: development of novel repellents Abstract The spread of blacklegged ticks (Ixodes scapularis) and growing threat of Lyme disease transmission has increased demand for effective, safe and environmentally friendly repellent products. Plant-derived essential oils are natural products that exhibit insecticidal and repellant activities and represent a promising alternative to synthetic repellants. However, mechanisms by which ticks detect odor stimuli and how such stimuli may function as repellents are not well understood. We examined the repellent activity of selected essential oil components towards I. scapularis in short- and long-term dose–response trials. To determine the specific olfactory organs involved in detection of chemical stimuli, we tested tick behavioral response in repellency bioassays after removing appendages that house chemosensory sensilla (e.g., foretarsi or pedipalps). New prototype formulae were tested in longevity trials repelling up to 95% of tested ticks after 1 h post-application. This study provides new insight regarding tick olfaction and behavior, and innovative methods for selecting appropriate chemicals for development of novel plant-based repellent products for protection from ticks.

Prevalence and molecular characterization of Rickettsia spp. in questing ticks from north-western Spain Abstract Tick-borne rickettsioses, most of them belonging to the spotted fever group (SFG), have been recognized as important emerging vector-borne zoonotic diseases. In order to determine the presence of Rickettsia spp. in questing ticks from north-western Spain, 1056 Ixodes ricinus, 19 Dermacentor marginatus, 17 Dermacentor reticulatus and one Ixodes acuminatus were processed. Rickettsia DNA was detected by PCR targeting rOmpA and rOmpB genes. A total of 219 (20.7%) I. ricinus, 19 (100%) D. marginatus and four D. reticulatus (23.5%) were positive. The prevalence was significantly higher in I. ricinus from coastal areas and in winter. Five species were identified: Rickettsia felis, Rickettsia monacensis, Rickettsia raoultii, Rickettsia slovaca and “Candidatus Rickettsia rioja”. Our results reveal a significant presence of some pathogenic Rickettsia species in questing tick populations from this area which involves a noticeable risk of rickettsiosis. As R. raoultii, R. slovaca and “Ca. R. rioja” DNA were identified in I. ricinus, considered an unusual vector for these Rickettsia species, further studies are needed to unravel the role of that tick species in the maintenance and transmission of these three Rickettsia species in north-western Spain.