Biochemical analysis of Hyalomma dromedarii salivary glands and gut tissues using SR-FTIR micro-spectroscopy.

Ticks are obligatory voracious blood feeders infesting diverse vertebrate hosts, that have a crucial role in the transmission of diverse pathogens that threaten human and animal health. The continuous emergence of tick-borne diseases due to combined worldwide climatic changes, human activities, and acaricide-resistant tick strains, necessitates the development of novel ameliorative tick control strategies such as vaccines. The synchrotron-based Fourier transform infrared micro-spectroscopy (SR-FTIR) is a bioanalytical microprobe capable of exploring the molecular chemistry within microstructures at a cellular or subcellular level and is considered as a nondestructive analytical approach for biological specimens. In this study, SR-FTIR analysis was able to explore a qualitative and semi-quantitative biochemical composition of gut and salivary glands of Hyalomma dromedarii (H. dromedarii) tick detecting differences in the biochemical composition of both tissues. A notable observation regarding Amide I secondary structure protein profile was the higher ratio of aggregated strands in salivary gland and beta turns in gut tissues. Regarding the lipid profile, there was a higher intensity of lipid regions in gut tissue when compared to salivary glands. This detailed information on the biochemical compositions of tick tissues could assist in selecting vaccine and/or control candidates. Altogether, these findings confirmed SR-FTIR spectroscopy as a tool for detecting differences in the biochemical composition of H. dromedarii salivary glands and gut tissues. This approach could potentially be extended to the analysis of other ticks that are vectors of important diseases such as babesiosis and theileriosis.

Systemic evaluation of novel acaricide hexythiazox for bioactivity improvement and risk reduction at the enantiomer level.

Traditional risk assessments of chiral pesticides mainly depend on racemic form, which is often incomprehensive. This study conducted systemic investigations on the bioactivity, toxicity, and ecotoxicological effects of hexythiazox (HTZ) at the enantiomer level. The elution order and absolute configuration of HTZ enantiomers were determined. (4R, 5R)-(+)-HTZ exhibited 708 and 1719 times higher bioactivity against Tetranychus cinnabarinus and Tetranychus urticae eggs than (4S, 5S)-(-)-HTZ, respectively. Molecular docking indicated greater interactions between (4R, 5R)-(+)-HTZ and chitin synthase leading to higher bioactivity of (4R, 5R)-(+)-HTZ. However, (4S, 5S)-(-)-HTZ induced greater changes in protein and malondialdehyde content, and antioxidant and detoxification enzyme activities than (4R, 5R)-(+)-HTZ in earthworms. Furthermore, integrated biomarker response results indicated (4S, 5S)-(-)-HTZ exhibited higher toxic effects on earthworms than (4R, 5R)-(+)-HTZ. Finally, significant differentially expressed genes (DEGs) were observed in earthworms after exposure to (4R, 5R)-(+)-HTZ and (4S, 5S)-(-)-HTZ, respectively. These DEGs were mainly enriched in glycolysis/gluconeogenesis and purine metabolism pathways in earthworms. Additionally, six metabolism pathways were also enriched, including pyruvate metabolism, fatty acid biosynthesis, oxidative phosphorylation, citric acid cycle, fatty acid degradation, and ATP-binding cassette transporters. These findings suggest that earthworms exhibited enantiomer-specific responses to (4R, 5R)-(+)-HTZ and (4S, 5S)-(-)-HTZ. This study provides systemic insight into the toxicity mechanism of HTZ at the enantiomer level and the potential to develop (4R, 5R)-(+)-HTZ as a high-efficiency and low-risk pesticide.

Discovery of Pesticide Candidates from Natural Plant Products: Semisynthesis and Characterization of Andrographolide-Based Esters and Study of Their Pesticidal Properties and Toxicology.

To explore the use of nonfood plant-derived secondary metabolites for plant protection, a series of ester derivatives for controlling the major migratory agricultural pests were obtained by structural modification of andrographolide, a labdane diterpenoid isolated from Andrographis paniculata. Compound Id showed good insecticidal activity against the fall armyworm Spodoptera frugiperda Smith. Compounds IIa (LC50: 0.382 mg/mL) and IIIc (LC50: 0.563 mg/mL), the acaricidal activities of which were, respectively, 13.1 and 8.9 times that of andrographolide (LC50: 4.996 mg/mL), exhibited strong acaricidal and control effects against Tetranychus cinnabarinus Boisduval. Against Aphis citricola Van der Goot, compounds IIIc and IVb displayed 3.9- and 3.7-fold pronounced aphicidal activity of andrographolide. Effects of compound Id on three protective enzymes (superoxide dismutase, peroxidase, and catalase) of S. frugiperda were also observed. The obvious differences of epidermal cuticle structures of mites treated with compound IIa were determined by scanning electron microscopy. Structure-activity relationships indicated that 14-ester derivatives of andrographolide showed potential insecticidal/acaricidal activities and can be further utilized as lead compounds.

Scabies: Immunopathogenesis and pathological changes.

Scabies is an itchy skin disease caused by the burrowing mite Sarcoptes scabiei. During their lifespan, the female mites invade the stratum corneum and create tunnels, in which they reside, move, feed, deposit fecal pellets, and lay eggs. Recently, scabies was included in the World Health Organization roadmap for neglected tropical diseases 2021-2030. This review attempts to summarize our knowledge about the mite's biology and the disease pathogenesis, pathological changes, and complications. Generally, the host-parasite interaction in scabies is highly complex and involves different mechanisms, some of which are yet largely unknown. Elucidation of the nature of such interaction as well as the underlying mechanisms could allow a better understanding of the mite's biology and the development of novel diagnostic and therapeutic options for scabies control programs. Moreover, identification of the molecular basis of such interaction could unveil novel targets for acaricidal agents and vaccines.

Construction and Single-Crystal Structures of N-Isoxazolin-5-ylcarbonylindole Derivatives, and Their Pesticidal Activities and Toxicology Study.

To explore natural product-based pesticide candidates, a series of indole derivatives containing the isoxazoline skeleton at the N-1 position were synthesized by 1,3-dipolar [2 + 3] cycloaddition reaction. Their structures were characterized by melting points (mp), infrared (IR) spectra, proton nuclear magnetic resonance spectra (1H NMR), carbon-13 nuclear magnetic resonance spectra (13C NMR), and high resolution mass spectrometry (HRMS). The single-crystal structures of five compounds were presented. Against Tetranychus cinnabarinus Boisduval, compound 3b showed greater than 3.8-fold acaricidal activity of indole and good control effects under glasshouse conditions. Against Aphis citricola Van der Goot, compounds 3b and 3q exhibited 48.3- and 36.8-fold aphicidal activity of indole and 6-methylindole, respectively. Particularly, compound 3b showed good bioactivities against T. cinnabarinus and A. citricola. Against Eriosoma lanigerum Hausmann, compound 3h and 3i showed 2.1 and 1.9 times higher aphicidal activity compared to indole. Furthermore, the construction of the epidermal cuticle layer of 3b-treated carmine spider mites was distinctly damaged, which ultimately led to their death.

Identification and Functional Characterization of an Omega-Class Glutathione S-Transferase Gene PcGSTO1 Associated with Cyetpyrafen Resistance in Panonychus citri (McGregor).

Cyetpyrafen is a recently developed acaricide. The citrus red mite, Panonychus citri (McGregor), has developed significant resistance to cyetpyrafen. However, the molecular mechanism underlying the cyetpyrafen resistance in P. citri remains unclear. Glutathione S-transferases (GSTs) play a critical role in arthropod pesticide resistance. This study showed that GSTs were potentially related to the resistance of P. citri to cyetpyrafen through synergistic experiments and enzyme activity analysis. An omega-family GST gene, PcGSTO1, was significantly up-regulated in the egg, nymph, and adult stages of the cyetpyrafen-resistant strain. Additionally, silencing of PcGSTO1 significantly increased the mortality of P. citri to cyetpyrafen and recombinant PcGSTO1 demonstrated the ability to metabolize cyetpyrafen. Our results indicated that the overexpression of PcGSTO1 is associated with cyetpyrafen resistance in P. citri, and they also provided valuable information for managing resistance in P. citri.

Some essential oils as potential control agents for varroa mite (Varroa destructor) in infected honey bees (Apis mellifera).

Background: Ecto-parasite, varroa mite, (Varroa destructor), is the primary pest affecting the apiculture sector globally in various regions. Aim: This study examined the toxicity of nine essential oils to Apis mellifera L. and the acaricidal impact of those oils against V. destructor. Methods: The acaricidal effects of nine essential oils, extracted from plant materials were used. In the screening experiment, 10 mg of the active ingredients of the plant material extracts were prepared in an alcohol solution with concentrations of 5%, 10%, and 15%. For each type of plant extract, five female V. destructor were transferred to a Petri dish with five worker bees incubated at 70% humidity and 33°-34° for 2 days, for each treatment four replicates were used compared to the control. Forty-eight hours following treatment, the number of dead and live mites was counted to determine the mortality rate. In the second assay experiment, the best five essential oils of the previous experiment were selected to re-assess their effectiveness on varroa mites and honeybee workers by using a concentration of 15%. Five females of V. destructor were transferred to a Petri dish with 10 adult bees and treated with the solution of the selected oils. Five replicates and control treatments were taken for each sample simultaneously. Dead and live bees were counted for each replicate at 48 hours after treatment. Results: There were no significant differences between the concentrations used of each oil on the rate of death of mites, and its effectiveness ranged between 70.0% and 53.3% compared to the control groups. In addition, the best oil used was bitter melon, with a death rate of 80% at a concentration of 15%, while peppermint oil showed the lowest death rate of 45% at a concentration of (5%). However, all these treatments were statistically highly significant compared with the natural death rate in control (2%). In the second test, the results of the statistical analysis indicated that there were highly significant differences (P0.05 <0.0001) in the average numbers of dead varroa mites compared to the control when using a 15% concentration of five selected oils. On the other hand, there was no statistically significant difference in the honey bee workers' mortality rate between the treatment and control groups (P0.05 <0.3390), and it was relatively low for all treatments except the basil oil, where the bee mortality rate was 16% compared to the control (10%). Conclusion: It is clear from this experiment that bitter melon oil can be used to control varroa mites and it can be considered safe for honey bees as well as for the environment.

Spatial distribution of two acaricides and five neonicotinoids in beehives and surrounding environments in China.

Managed bees commonly suffer from cross-contamination with acaricides and neonicotinoids, posing robust threats to bee population health. However, their residual characteristics and spatial distribution in beehives and surrounding environments are poorly understood. This study detected two common acaricides and five neonicotinoids in 240 beehive samples and 44 surrounding environmental samples collected from 25 Chinese provinces. The results showed that 40.0% of the honey samples contained acaricides and 83.1% contained neonicotinoids. Neonicotinoid concentrations in honey were geographically distinguished by the "Hu Huanyong line", and concentrations of neonicotinoids in honey from eastern areas were 2.65-fold higher than those in honey from western areas. Compared to the approved acaricide amitraz, the banned acaricide coumaphos was detected more frequently in honey and was positively correlated with that quantified in the paired pollen samples. Although coumaphos was identified in only three soil samples, lower coumaphos residues in honey might be associated with persistent pollution in the surrounding environment. Conversely, neonicotinoids were detected at higher levels in honey than in the pollen and soil, demonstrating that the neonicotinoid residues in honey have a cumulative effect. This study contributes to a better understanding of the pesticide contamination scenarios that underlie the exposure risks of bees.

Alpha- and beta-pinene isomers act differently to control Rhipicephalus microplus (Acari: Ixodidae).

The cattle tick Rhipicephalus microplus is an ectoparasite of high importance in veterinary medicine and public health. Since synthetic chemicals used to control these ticks can select resistant strains and cause toxic effects in their hosts, there is a need to identify effective substances with fewer adverse effects. For this reason, we investigated the effects of alpha- and beta-pinene, known for their various biological effects, on the mortality and reproductive performance of R. microplus engorged female ticks. The products were diluted in a 2% Tween 80 aqueous solution. The ticks were first weighed and then immersed in the test solutions for five minutes. Then, they were dried with paper towels and fixed dorsoventrally in Petri dishes, totalling five treatment groups for each pinene and a control group treated with the solvent alone. The ticks were monitored daily for mortality, and their eggs were collected and weighed. The larval hatching rate was estimated, and the pre-oviposition and incubation periods were determined. From these data, the following parameters were calculated: egg production index, fertility rate, estimated reproduction rate, percentages of reduction in oviposition and hatching, and product efficacy. Alpha-pinene showed better results at higher concentrations, unlike beta-pinene, which was more effective at lower concentrations. The effectiveness of alpha-pinene was 74% at a concentration of 14.0 µL/mL, while beta-pinene showed 78% efficacy at 2.0 µL/mL. The results indicated for the first time different effects of two isomers in ticks, suggesting that these compounds act on R. microplus females in different ways.

Evaluation of the efficacy of Simparica Trio® in the prevention of the transmission of Babesia canis by infected Dermacentor reticulatus to dogs.

BACKGROUND: Babesia canis is a clinically relevant vector-borne pathogen in dogs, and its presence is expanding. The efficacy of Simparica Trio® (Zoetis) in the prevention of B. canis transmission was evaluated at the minimum recommended label dose of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5 mg/kg pyrantel per kg bodyweight. METHODS: Twenty-four (24) dogs were randomly allocated to either a placebo-treated group or one of two treatment groups with Simparica Trio. Dogs were infested with B. canis-infected Dermacentor reticulatus ticks 21 or 28 days after treatment administration. Blood samples for antibody and DNA detection were collected from each dog prior to tick infestation until 28 days after infestation. A dog was defined as being B. canis positive if it tested positive by both an indirect immunofluorescence assay (IFA) and PCR at any time during the study. RESULTS: No treatment-related adverse reactions were recorded during the study. All placebo-treated animals displayed clinical signs due to babesiosis and tested positive on both IFA and PCR. None of the Simparica Trio-treated animals displayed any clinical symptoms or tested positive, resulting in a 100% efficacy in the prevention of canine babesiosis (P < 0.0001). CONCLUSIONS: A single treatment with Simparica Trio at the minimum recommended label dose of 1.2 mg/kg sarolaner, 24 µg/kg moxidectin and 5 mg/kg pyrantel per kg bodyweight prevents the transmission of B. canis by infected D. reticulatus to dogs for at least 28 days.

Relationship between acaricide resistance and acetylcholinesterase gene polymorphisms in the cattle tick Rhipicephalus microplus.

In this study, we aimed to develop a comprehensive methodology for identifying amino acid polymorphisms in acetylcholinesterase transcript 2 (AChE2) in acaricide-resistant Rhipicephalus microplus ticks. This included assessing AChE2 expression levels through qPCR and conducting 3D modeling to evaluate the interaction between acaricides and AChE2 using docking techniques. The study produced significant results, demonstrating that acaricide-resistant R. microplus ticks exhibit significantly higher levels of AChE expression than susceptible reference ticks. In terms of amino acid sequence, we identified 9 radical amino acid substitutions in AChE2 from acaricide-resistant ticks, when compared to the gene sequence of the susceptible reference strain. To further understand the implications of these substitutions, we utilized 3D acaricide-AChE2 docking modeling to examine the interaction between the acaricide and the AChE2 catalytic site. Our models suggest that these amino acid polymorphisms alter the configuration of the binding pocket, thereby contributing to differences in acaricide interactions and ultimately providing insights into the acaricide-resistance phenomenon in R. microplus.

Title: Relations entre la résistance aux acaricides et les polymorphismes du gène de l'acétylcholinestérase chez la tique du bétail Rhipicephalus microplus. Abstract: Notre étude vise à développer une méthodologie complète pour identifier les polymorphismes d'acides aminés dans le transcrit 2 de l'acétylcholinestérase (AChE2) chez les tiques Rhipicephalus microplus résistantes aux acaricides. Cela comprend l'évaluation des niveaux d'expression d'AChE2 via qPCR et la réalisation d'une modélisation 3D pour évaluer l'interaction entre les acaricides et l'AChE2 à l'aide de techniques d'amarrage moléculaire. L'étude a produit des résultats significatifs, démontrant que les tiques R. microplus résistantes aux acaricides présentent des niveaux d'expression d'AChE significativement plus élevés que les tiques sensibles de référence. En termes de séquence d'acides aminés, nous avons identifié 9 substitutions d'acides aminés dans AChE2 provenant de tiques résistantes aux acaricides par rapport à la séquence génétique de la souche sensible de référence. Pour mieux comprendre les implications de ces substitutions, nous avons utilisé la modélisation de l'amarrage acaricide-AChE2 pour examiner l'interaction entre l'acaricide et le site catalytique AChE2. Nos modèles suggèrent que ces polymorphismes d'acides aminés modifient la configuration de la poche de liaison, contribuant ainsi aux différences dans les interactions acaricides et fournissant finalement un aperçu du phénomène de résistance aux acaricides chez R. microplus.

Properties of essential oils absorbed on the surface of cardboard pieces after using atmospheric-pressure plasma treatments to develop long-lasting Varroa miticides in honeybees (Apis mellifera).

The ectoparasitic mite, Varroa destructor is the most serious widespread pest of managed honeybees (Apis mellifera). Several acaricide products, which include essential oils, have been proposed for mite control. In this study, we aimed to apply atmospheric-pressure plasma to modify a cardboard piece surface in order to prolong the delivery of essential oils for controlling Varroa in honeybee colonies. Absorption capacity, release rates and evaporation rates of essential oils were determined. Cardboard piece showed a higher absorption capacity of cinnamon compared to citronella and clove. Surface modification of cardboard pieces using argon plasma at different gas flow rates and treatment durations, significantly affected the absorption of clove oil. Additionally, the release rate of cinnamon, citronella and clove was significantly enhanced after argon plasma treatments. Evaporation of cinnamon was dramatically increased by plasma treatment at 6-h of incubation. The highest evaporation rate was obtained by plasma-treated cardboard piece at a gas flow rate of 0.5 Lpm for 60 s (0.2175 ± 0.0148 µl/g•h). Efficiency of plasma-treated cardboard piece, impregnated with essential oils, was also investigated for Varroa control in honeybee colonies. In the first experiment, formic acid 65% (v/v) showed the highest efficiency of 90.60% and 81.59% with the percent of mite infestation was 0.23 ± 0.13% and 0.47 ± 0.19% at 21 and 35 days, respectively after treatment. The efficacy of cardamon oil (5% (v/v)) delivered using plasma-treated cardboard pieces was 57.71% (0.70 ± 0.16% of mite infestation) at day 21 of experiment. However, the delivery of cardamon oil at the concentration of 1% and 5% (v/v) by untreated cardboard piece had 16.93% and 24.05% of efficacy to control mites. In the 2nd experiment, the application of plasma-treated cardboard pieces impregnated with 5% (v/v) clove oil induced a 38.10% reduction in the population of Varroa mites followed by 5% (v/v) of cardamon with 30% efficiency. Although, the infestation rate of Varroa in colonies was not significant different between treatments, essential oils delivered using plasma-treated cardboard pieces tended to decrease Varroa population in the treated colonies. Hence, atmospheric-pressure plasma for the modification of other materials, should be further investigated to provide alternative control treatment applications against honeybee mites.

Larvicidal and repellent effects of essential oils on the brown dog tick (Rhipicephalus sanguineus Sensu lato) with description of new larval repellent activity test method.

The aim of this research was to investigate the larvicidal and repellent effects of essential oils (EOs) obtained from two Lamiaceae plant species, Origanum minutiflorum O. Schwarz & P.H. Davis and Dorystoechas hastata Boiss. & Heldr. ex Bentham, both endemic to Turkey, on Rhipicephalus sanguineus s.l. Latreille (Acari: Ixodidae). The study also introduces a new test method that can be used to assess the repellent effects against ticks. Both plant EOs exhibited the highest larvicidal activity against brown dog tick larvae after 24 h and LC50 and LC90 values were determined as 0.101% and 0.125% for O. minutiflorum essential oil and 0.937% and 2.1% for D. hastata essential oil, respectively. In this study, we have described a detailed protocol for a novel larval repellent activity test (LRAT) for essential oils and extracts, using simple equipment. The advantages and limitations of LRAT, when compared to other tests commonly used to determine repellent effect against ticks, are also included in this study. The LRAT was developed with modifications of the larval immersion test (LIT) and proves to be a highly efficient and easily observable method. It can be used to test any active substance that may be toxic to humans and animals. According to the LRAT, at the end of 3 h, O. minutiflorum essential oil showed a high repellent effect, varying between 84.14% and 100% at 1% concentration. This result was not statistically different from the DEET, the positive control. When comparing the larvicidal and repellent activities, O. minutiflorum essential oil was found to be more effective than D. hastata essential oil.

Subchronic exposure to fenpyroximate causes multiorgan toxicity in Wistar rats by disrupting lipid profile, inducing oxidative stress and DNA damage.

BACKGROUND: Fenpyroximate (FEN) is an acaricide that inhibits the complex I of the mitochondrial respiratory chain in mites. Data concerning mammalian toxicity of this acaricide are limited; thus the aim of this work was to explore FEN toxicity on Wistar rats, particularly on cardiac, pulmonary, and splenic tissues and in bone marrow cells. METHODS: rats were treated orally with FEN at 1, 2, 4, and 8 mg/Kg bw for 28 days. After treatment, we analyzed lipid profile, oxidative stress and DNA damage in rat tissues. RESULTS: FEN exposure increased creatinine phosphokinase (CPK) and lactate dehydrogenase (LDH) activities, elevated total cholesterol (T-CHOL), triglycerides (TG), and low-density lipoprotein cholesterol (LDL-C) concentrations, while decreasing high-density lipoprotein cholesterol (HDL-C). It inhibited acetylcholinesterase (AChE) activity, enhanced lipid peroxidation, protein oxidation, and modulated antioxidant enzymes activities (superoxide dismutase, catalase, glutathione peroxidase, and glutathione S-transferase). Comet assay indicated that FEN induced a dose-dependent DNA damage, contrasting with the micronucleus test showing no micronuclei formation. Nonetheless, FEN exhibited cytotoxicity to bone marrow cells, as evidenced by a reduction in the number of immature erythrocytes among total cells. CONCLUSION: FEN appears to carry out its genotoxic and cytotoxic activities most likely through an indirect pathway that involves oxidative stress.

What is the optimal timing to initiate strategic control of Rhipicephalus microplus in taurine cattle in a tropical region?

Cattle tick control poses a significant challenge for livestock in tropical and subtropical regions. The objective of this study was to determine the most suitable timing to initiate a strategic tick control program and to identify the ideal number of acaricide treatments for adult taurine cattle (Bos taurus taurus) in a tropical region throughout the year. Three groups with 10 bovines each were performed: T01 (strategic treatment in late autumn/winter/late spring, every 28 days), T02 (strategic treatment to act in the "first tick generation" - early spring/summer/early autumn, every 28 days) and T03 (control). Tick counts (females 4-8 mm) were conducted every 14 days. If the tick burden in any group reached 30 or more during these counts, we applied an additional treatment. Over the course of a year, T02 required significantly fewer (p < 0.05) acaricide treatments than T01, with nine treatments for T02 and eleven for T01. Furthermore, during the tick counts, animals in T02 showed a lower tick burden compared to those in T01. Initiating the strategic tick control program in early spring, corresponding to the first tick generation, proved more effective than starting in autumn. This approach not only required fewer acaricide treatments but also resulted in a reduced tick burden. These benefits are particularly valuable in terms of animal welfare and managing acaricide resistance issues.

Polymeric films of corn starch enhance the lethal effects of thymol and carvacrol terpenes upon Rhipicephalus microplus ticks.

The tick Rhipicephalus microplus is a parasite of great importance in cattle breeding. It is responsible for huge economic losses. The application of synthetic acaricides is used as a form of control. However, resistant strains have been selected over the years, making it necessary to search for new alternative formulations. The present study aimed to formulate biodegradable films impregnated with the terpenes carvacrol and thymol and evaluate their efficacy on larvae and adults of R. microplus through in vitro tests. The following formulations were prepared: Film 1 (starch based); Film 2 (based on starch and glycerol); Film 1 + Carvarcol or Thymol; Film 2 + Carvarcol or Thymol. Terpenes had a final concentration of 5.0 mg/mL. To evaluate the formulations on larvae, the immersion test was performed by dividing into six groups according to the concentration of terpenes: 5.0, 2.5, 1.25, 0.625, 0.313, 0.156 mg/mL and the control groups: 1% ethanol solution; 10% ethanol solution; Film 1; and Film 2. For the evaluations on adult ticks, ten experimental groups (n = 10) were used: 1) Carvacrol; 2) Film 1 + Carvacrol; 3) Film 2 + Carvacrol; 4) Thymol; 5) Film 1 + Thymol; 6) Film 2 + Thymol; 7) Distilled water; 8) 10% ethanol solution; 9) Film 1; and 10) Film 2. In experimental groups 1-6, carvacrol and thymol (free or incorporated in two different biodegradable film formulations) were evaluated at the same concentration (5.0 mg/mL). Each group of ticks was immersed in their respective solutions for five minutes. The results of the tests on larvae showed that the Film 1 + thymol and Film 2 + carvacrol formulations had the lowest lethal concentrations (0.076 and 0.255 mg/mL, respectively), values up to 9.0-fold lower than the monoterpenes tested outside the formulation. Carvacrol and thymol at the concentrations tested were effective in controlling engorged females with a percentage of 32.2% and 63.8%, respectively. When incorporated into biodegradable film formulations, these monoterpenes showed much greater efficacy. Film 1 + carvacrol and Film 2 + carvacrol with control percentages of 71.6% and 97.2%, respectively, while the formulations Film 1 + thymol and Film 2 + thymol showed values of 96.9% and 100.0%. The tick control activity of the biopolymer formulations with thymol and carvacrol was demonstrated through the high mortality rates of larvae and engorged females of the tick R. microplus. Therefore, the results obtained indicate that these formulations have great potential for tick control mainly because of the percentage of control up to 100% in engorged females in in vitro tests.

Two protocols using fluralaner for Rhipicephalus microplus strategic control on taurine cattle in a tropical region.

BACKGROUND: The present study aimed to evaluate the effects of different treatment strategies using a new commercial formulation containing pour-on fluralaner on Rhipicephalus microplus infestation in cattle and in pastures in a tropical climate region where up to five generations of this tick species can occur per year. METHODS: Forty-five naturally infested cattle were divided into three experimental groups: T01, treated with fluralaner (2.5 mg/kg) pour-on every 42 days; T02, the cattle received the first treatment with fluralaner on Day 0 but the next treatment involved a weekly visual evaluation; T03, control, received palliative treatment with a spray formulation when the group mean was ≥ 30 ticks. Counts of female R. microplus were performed weekly until day 343, and larval counts on pasture were performed on Days 0, 30, and 60 and every 30 days until Day 330. RESULTS: Using fluralaner, six applications were performed in Group T01, and four were performed in Group T02. In the control group (T03), it was necessary to perform eight palliative acaricide treatments with the spray formulation. The animals in T01 and T02 showed lower mean tick counts (p ≤ 0.05) than the control group (T03) on 28 and 27 of the 49 evaluated dates, respectively. In the paddock where the animals were kept as controls, the R. microplus larvae counts increased to 1458. In the paddocks where the animals were treated with fluralaner, the number was ≤ 19 per paddock during the study. CONCLUSIONS: The different strategic treatment protocols performed with pour-on fluralaner (2.5 mg/kg) over a year in taurine cattle in a tropical region with a history of up to five annual generations of cattle ticks were effective, maintaining levels of R. microplus infestations in animals and in pastures close to zero in most of the study. Depending on the retreatment criterion adopted, the number of applications per year may be lower, resulting in a reduction in the mean cost of acaricide treatment per year and lower exposure of R. microplus populations to the active ingredient, resulting in lower resistance and selection pressure.

Molecular identification of hemoparasites in ixodid ticks in Iran.

A total of 386 ticks were processed in order to investigate the occurrence of selected tick transmitted pathogens (i.e., Theileria, Babesia, Hepatozoon and Cytauxzoon) in ixodid ticks in six provinces of Iran (East Azerbaijan, Gilan, Kermanshah, Khuzestan, Sistan & Baluchestan and Tehran). Ticks identified as Dermacentor marginatus, Hyalomma aegyptium, Hy. anatolicum, Hy. asiaticum, Hy. marginatum, Ixodes ricinus, Rhipicephalus annulatus and R. sanguineus sensu lato were collected from sheep and cattle. Conventional PCR and sequencing results revealed DNA of Theileria ovis in three R. sanguineus sensu lato pools and one D. marginatus pool from sheep in Kermanshah and East Azerbaijan, T. annulata in one Hy. asiaticum pool from cattle in Kermanshah, and He. canis in an individual female Hy. aegyptium in Kermanshah. Data herein indicate the role of R. sanguineus complex and D. marginatus in the epidemiology of ovine theileriosis in western and northwestern Iran. Potential role of Hyalomma aegyptium in the transmission of He. canis is discussed. Considering non-principled movement of livestock across the country and increasing reports about the resistance of ticks to common acaricides, test-and-treatment of infected livestock, vaccination of the livestock against endemic tick-borne pathogens, and the use of non-chemical tick management strategies are recommended.

Aural hematoma in lambs associated with Otobius megnini (Ixodida: Argasidae) infestation.

In this report we described a case of aural hematomas in three lambs associated with Otobius megnini (Ixodida: Argasidae) infestation. From April to May 2021, five 3-month-old Hampshire cross lambs presented with unilateral aural hematomas. Upon otoscopic examination, engorged soft ticks (O. megnini) were observed in the external ear canals of three of the five lambs. The remaining two lambs had lesions consistent with infestation and were in a shared environment and deemed likely to have been infected. The treatment of all animals was based on the drainage of the serosanguinous fluid through an incision in the internal space of the ear pinna. Upon physical inspection of the entire flock (n = 310), O. megnini infestation was observed in one additional animal that did not have a hematoma. Following animal and environmental ectoparasiticide treatment with permethrin, no recurrences or additional cases of aural hematomas were observed in the flock in the following two-year period. To the authors' knowledge, this is the first description of aural hematomas in lambs associated with O. megnini infestation with successful recovery after surgery and off-label acaricide treatment.

The Varroa paradox: infestation levels and hygienic behavior in feral scutellata-hybrid and managed Apis mellifera ligustica honey bees.

The Varroa destructor mite is a parasitic threat to managed and feral honey bee colonies around the world. Beekeepers use miticides to eliminate Varroa in commercial hives, but these chemicals can diminish bee health and increase miticide resistance. In contrast, feral honey bees have developed multiple ways to counteract mites without chemical treatment. We compared mite levels, grooming habits, and mite-biting behavior between feral Africanized honey bees (genomically verified Apis mellifera scutellata hybrids) and managed Italian honey bees (A. mellifera ligustica). Surprisingly, there was no difference in mite infestation levels between scutellata-hybrids and managed bees over one year despite the regular use of miticides in managed colonies. We also found no differences in the social immunity responses of the two groups, as measured by their hygienic habits (through worker brood pin-kill assays), self-grooming, and mite-biting behavior. However, we provide the first report that both scutellata-hybrids and managed honey bees bite off mite chemosensory forelegs, which the mites use to locate brood cells for reproduction, to a significantly greater degree than other legs (a twofold greater reduction in foreleg length relative to the most anterior legs). Such biting may impair mite reproduction.