Histopathological effects of the fruit extract of Citrullus colocynthis on the ovary of the tick Hyalomma dromedarii.

Hyalomma dromedarii is the predominant tick species parasitizing camels in Egypt which leads to mortalities in young animals that result in economic losses. It can transmit a lot of pathogens to animals and humans, such as the Crimean-Congo hemorrhagic fever virus, the Dhori virus, Kadam virus, Theileria annulata and spotted fever rickettsia. The continuous use of chemical acaricides has negative impact on the environment and almost led to acaricidal resistance, and hence the plant extracts represent alternative methods for controlling ticks. The present study was carried out to assess the histopathological effects on the ovary of fed female Hyalomma dromedarii following immersion in the ethanolic extract of fruits of Citrullus colocynthis (100 mg/mL). Light, scanning and transmission electron microscopy observations provided evidence that Citrullus colocynthis caused extensive damage to oocytes. Destruction of the internal organelles of oocytes, along with delay and/or inhibition of vitellogenesis were demonstrated. This is the first histological study that points to damage in H. dromedarii ovaries following treatment with the ethanolic extract of fruits of C. colocynthis. The data presented suggest that the plant extract affects the ovary either directly by entering the oocytes and/or indirectly by damaging the gut cells and digestion of blood that interfere with the development of oocytes, so it can be used as a promising agent for tick control.

Surveillance of ticks on a tropical island with introduced ungulates.

The abundance of hard ticks can be determined by abiotic factors and the presence of suitable hosts. Since deer represent important hosts for many tick species, it is imperative to investigate whether deer introduction will increase the number of ticks, which can transmit a wide variety of pathogens. The sika deer (Cervus nippon) was introduced to Green Island, Taiwan, to supply velvet antlers, a traditional Chinese medicine. However, they were later released into the wild after a steep decline in antler prices. We conducted surveys for questing ticks, ticks on rodents and shrews, and the fecal pellet groups of ungulates (deer and goats) in 31 transects within 3 habitat types (forest, grassland, and roadside) on Green Island every 2 months in 2019. A total of 5,321 questing ticks were collected. All collected ticks were Haemaphysalis mageshimaensis Saito and Hoogstraal, a species first identified on an island with an abundance of sika deer. Additionally, 48 ticks collected from 126 trapped rodents and shrews were almost invariably Ixodes granulatus (except for 1 larval Haemaphysalis sp.). We did not find a positive correlation between the number of questing ticks and the number of fecal pellet groups, suggesting that factors other than ungulate abundance affect tick abundance. Nevertheless, large populations of questing ticks along some roadsides suggest a high risk of tick-borne diseases to tourists on this tropical island.

Repellent effects of Chinese cinnamon oil on nymphal ticks of Haemaphysalis longicornis, Rhipicephalus haemaphysaloides, and Hyalomma asiaticum.

The repellent activity of Chinese cinnamon oil (Cinnamomum cassia) on nymphal ticks (Haemaphysalis longicornis Neumann, Rhipicephalus haemaphysaloides Supino, and Hyalomma asiaticum Schulze and Schlottke) was evaluated in a sample Y-tube bioassay. The results were based on the vertical migration of ticks during the host-seek phase and showed a dose-dependent repellent effect of Chinese cinnamon oil on the tested nymphs after 6 h. For H. longicornis, R. haemaphysaloides, and H. asiaticum at the concentrations (vol/vol) of 3, 3, and 1.5%, the repellent percentages over time were 68-97, 69-94, and 69-93%, respectively, which indicated strong repellent activities against ticks, similar to the positive control DEET (N,N-diethyl-3-methylbenzamide). Chinese cinnamon oil exerted the strongest effect on H. asiaticum nymphs. To our knowledge, this is the first study to investigate the repellent effects of Chinese cinnamon oil on ticks. Chinese cinnamon oil has considerable potential and should be developed as a practical tick repellent.

Novel acaricidal and growth-regulating activity of Aloe vera and Rheum rhabarbarum extracts and their oil/water nanoemulsions against the camel tick, Hyalomma dromedarii.

Hyalomma dromedarii is an important tick species infesting livestock. This work evaluated the novel adulticidal, insect growth-regulating, and enzymatic efficacy of ethanol plant extracts of Aloe vera and Rheum rhabarbarum and their nanoemulsions against males and engorged females of the camel tick, H. dromedarii. The physicochemical properties of nanoemulsions were evaluated. The High-Performance Liquid Chromatography (HPLC) analyses indicated that the extracts contained polyphenols and flavonoids, which could enhance their acaricidal effect. Dynamic light scattering (DLS) of the nanoemulsions of A. vera and R. rhabarbarum were 196.7 and 291 nm, whereas their zeta potentials were - 29.1 and - 53.1 mV, respectively. Transmission electron microscope (TEM) indicated that nanoemulsions showed a regular spherical shape (less than 100 nm). Fifteen days post-treatment (PT) with 25%, the mortality% of A. vera and R. rhabarbarum were 88.5 and 96.2%, respectively. Five days PT, the median lethal concentration values of A. vera, R. rhabarbarum, and their nanoemulsions were 7.8, 7.1, 2.8, and 1.02%, respectively, and their toxicity indices were 91.02, 100, 36.4, and 100%, respectively. Their median lethal time values PT with 3.5% were 6.09, 5.09, 1.75, and 1.34 days, respectively. Nanoemulsions enhanced the efficacy of the crude extract 1-7 folds, 5 days PT, and accelerated their speed of killing ticks 2-4 times. The total protein and carbohydrates, Acetylcholinesterase, Alpha esterase, and Amylase were affected PT. The reproductive potential of engorged females was adversely impacted. In conclusion, the novel A. vera and R. rhabarbarum extracts were promising acaricides, and their nanoformulations enhanced their efficacies.

Efficacy of hydroethanolic extract of Randia aculeata seed against the southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) on naturally infested cattle under field conditions.

The cattle tick Rhipicephalus (Boophilus) microplus is a major problem of concern for cattle industry in tropical and subtropical areas. Control of cattle tick is based mainly on the use of chemical acaricides, which has contributed to the emerging problem of selection of resistant tick lineages. Plants have been used as an alternative to conventional acaricidal drugs. On the other hand, the acaricidal activity of hydroethanolic extract of Randia aculeata seed (EHRA) has been demonstrated against R. microplus under laboratory conditions. However, the utility of EHRA seed as a potential acaricidal needs to be determined under field conditions. For this reason, the aim of this study was to evaluate the efficacy of the EHRA against R. microplus sprayed on naturally infested calves, determine the effect of the EHRA seed on acetylcholinesterase activity in R. microplus larval and identify the chemical composition of EHRA. Forty-five male calves were divided in three groups and treated with: G1 water; G2 EHRA 20% w/v and G3 coumaphos 0.2% v/v. Acetylcholinesterase (AChE) activity in R. microplus larvae was determined by a colorimetric assay. The chemical composition of EHRA was accessed through HPLC/MS. Significantly fewer ticks were observed after 24 h on the treated group compared to control group. EHRA significantly inhibited in vitro AChE activity in R. microplus at all tested concentrations. Chlorogenic acid, vanillinic acid, p-coumaric acid, caffeic acid. rutin, quercetin, (-)-epicatechin, 4-hydroxybenzoic acid, quercetin, vanillin, 2,4-dimethoxy-6-methylbenzoic acid, scopoletin and ferulic acid were identified in the extract. The results provided new data for the elucidation of the mechanisms of EHRA acaricide action and to further evaluate the use as a new alternative control agent against R. microplus under in vivo conditions.

Acaricidal activity of a 2-methoxy-clovan-9-ol rich fraction from Eugenia copacabanensis (Myrtaceae) extract on Rhipicephalus microplus (Acari: Ixodidae).

Rhipicephalus microplus (Ixodidae, canestrini, 1888) is an invasive ectoparasite of cattle which causes high economic losses in emerging countries such as Brazil. Phytochemical compounds have been tested as an alternative to synthetic acaricides due to potentially lower mammalian toxicity. This study evaluated the acaricidal activity against R. microplus of the 2-methoxy-clovan-9-ol rich fraction obtained from Eugenia copacabanensis Kiaersk.leaves, a little known Myrtaceae species from the Brazilian Atlantic Forest. This fraction was obtained by maceration with methanol, partition with n-hexane and purification by normal-phase column chromatography. GC-MS, 1H and 13C NMR and IR analysis contributed to the identification of a major compound as the sesquiterpene 2-methoxy-clovan-9-ol, reported for the first time for the Myrtaceae family. The fraction was tested against R. microplus unfed larvae and engorged females and a 93% larval mortality was observed at the concentration of 50 mg mL-1. Lower concentrations of the solution tested demonstrated a significant difference in egg mass weight, hatching and control percentage. Experiments with 50.0 mg mL-1 showed significative results, with lower concentration and maximum efficacy for both assays. The IC50 values for unfed larvae and engorged females were 21.76 and 11.13 mg mL-1, respectively. These results were similar to those obtained in other studies with isolated botanical compounds and essential oils. The lower IC50 for engorged females than for unfed R. microplus larvae had also been described for other plant materials, including plants from the same Myrtaceae family. The present result suggested different mechanisms of action of the compound on the reproductive biology of engorged females, improving its effect against egg viability. These results are important for tick control, suggesting that 2-methoxy-clovan-9-ol could be a potential natural acaricidal product against both R. microplus unfed larvae and engorged females.

Acaricidal and repellent activities of ethanol extracts of nine chinese medicinal herbs against Rhipicephalus microplus (Acari: Ixodidae).

Rhipicephalus microplus is a major threat to the cattle industry worldwide. The intensive use of acaricides and repellents has resulted in drug resistance. Hence, effective and eco-friendly pest control alternatives are urgently needed, especially from natural plant resources. In this study, the acaricidal and repellent activities of nine herbs against the larvae and eggs of R. microplus were evaluated. The results showed that ethanol extracts of star anise (Illicium verum), chaulmoogra (Hydnocarpus anthelmintica), motherwart (Leonurus artemisia), mandarin orange peel (citri reticulatae pericarpium, i.e., peel of Citrus reticulata fruit), and stemona (Stemona sessilifolia) had good contact acaricidal activities of 100, 98, 94, 88 and 86%, respectively, whereas star anise and clove (Syzygium aromaticum) had good fumigant acaricidal activities of 98 and 96%, respectively. The hatching inhibition rate of star anise against R. microplus eggs was 100%. All nine herbs had good real-time repellent rates, but only castor bean and star anise had repellent effects after 48 h (81.3 and 79.6%, respectively). This is the first report of the acaricidal and repellent activities of these medicinal herbs against R. microplus. Ethanol extracts of these herbs might be considered as potential alternatives to chemical acaricides for control of R. microplus.

Ixodicidal effect of extracts from Cordia boissieri, Artemisia ludoviciana and Litchi chinensis on Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The ixodicidal activity of the methanolic extracts of Artemisia ludoviciana (Astereceae), Cordia boissieri (Boraginaceae) and Litchi chinensis (Sapindaceae) against two field populations of Rhipicephalus (Boophilus) microplus from the state of Nuevo Leon (NL) and Veracruz (VER) was evaluated. The extract of L. chinensis in the concentration of 150 mg/ml showed efficacies of 100% and 99% against engorged females and mortalities of 98% and 99% against larvae. C. boissieri in the same concentration showed efficacies of 71% and 37% against engorged adults and mortalities of 33.04% and 10.33% against larvae and A. ludoviciana had efficacies of 94% and 83% in adults and mortalities of 89.39% and 89.21% against larvae in both populations respectively. The enzymatic activity of Acetylcholinesterase (AChE), Carboxylesterase (CaE), Glutathione-S-Transferase (GST) and Alkaline Phosphatase (ALP) was measured in both populations of ticks. As a result, a significant difference between both populations was shown, being the VER population the one that exhibited a higher enzymatic activity (p ≤ 0.05). It can be concluded that the methanolic extract of the seed of L. chinensis shows potential ixodicidal activity and can be used as an alternative source of tick control, however, prior characterization, toxicity and formulation studies are necessary.

Molecular characterization of Hyalomma dromedarii and evaluation of acaricidal potential of herbal methanolic extracts against H. dromedarii larvae in comparison to synthetic acaricides.

The present study had two aims: molecular characterization of Hyalomma dromedarii infesting one-humped camels of Haryana (North India), and assessment of the acaricidal potential of herbal methanolic extracts against H. dromedarii larvae in comparison to synthetic acaricides. Phylogenetics and population neutrality indices were assessed by targeting partial amplification of mitochondrial 16S rDNA sequences. Larval packet test (LPT) was performed to evaluate the acaricidal efficacy of herbal extracts (Ferula asafoetida and Trachyspermum ammi) and synthetic acaricides (deltamethrin and fipronil). Phylogenetic studies established the collected ticks to be H. dromedarii, exhibiting a homology of 99.8-100%. However, the present study isolates formed a different sub-clade compared to H. dromedarii sequences from Egypt, Senegal, Tunisia and Saudi Arabia. Nucleotide and haplotype diversity values were indicative of demographic expansion and low gene flow. Negative values of Tajima's D (-0.612) and Fu and Li's Fst (-0.479) highlighted deviations from neutrality and emphasized recent population expansion. The median lethal concentration (LC50) values recorded for T. ammi, F. asafoetida and their combination were 3.68, 2.87 and 2.59 mg/mL, respectively, whereas the 90% lethal concentration (LC90) values were 4.09, 3.58 and 3.35 mg/mL, respectively. It was also observed that the H. dromedarii population under study was completely susceptible to both the formulated synthetic acaricides. In conclusion, combination of methanolic extracts of F. asafoetida and T. ammi could provide a potential substitute to toxic synthetic chemical acaricides and might prove a valuable component of integrated tick management strategies.

In vitro acaricidal activity of Piper longum L. against amitraz resistant Rhipicephalus microplus (Acari: Ixodidae).

The cattle tick, Rhipicephalus microplus Canestrini (Acari: Ixodidae) is one of the most important tick species severely affecting health and causes huge losses to dairy industry. Chemical acaricides are mainly applied for tick control but development of resistance, environmental pollution and contamination of milk and meat products with residues has led to exploration alternative eco-friendly tick control strategies. The dried fruits of Piper longum L. (Indian long pepper, Thippali or Pippali) generally used as flavoring agent have also been shown to have insecticidal property. Different concentrations (0.625%-10%) of alcoholic and aqueous extracts of Piper longum L. were prepared and evaluated for acaricidal activity against amitraz resistant R. microplus adult and larval stages. Against larval stages a dose-dependent mortality response was recorded for both extracts and higher acaricidal property was exhibited by the alcoholic extract with LC50 and LC95 (95% CL) values of 0.488% (0.48-0.49) and 1.39% (1.35-1.44), respectively. Similarly, against adult engorged females, ethanolic extract showed higher acaricidal property with LC50 and LC95 (95% CL) values of 4.67% (4.61-4.74) and 12.38% (12.05-12.73), respectively. Significant (p < 0.05) reduction was recorded in reproductive index of ticks treated and but no effect on hatchability of eggs was recorded in treated groups. The present study establishes acaricidal activity of P. longum fruit extracts against both larval and adult stages of amitraz resistant population of cattle tick.

Acaricidal effect of Citrullus colocynthis fruit extract on the camel tick Hyalomma dromedarii (Koch, 1844).

The hard tick Hyalomma dromedarii is one of the most injurious ectoparasites affecting camels in Egypt and several other areas of the world. Hyalomma dromedarii engorged females were dipped in alcoholic extract of Citrullus colocynthis fruits at concentrations of 25, 50, 100, 200 and 400 mg/mL and then observed over four weeks following treatment. There was a marked increase in dead females following single treatment with the above concentrations, particularly for the two highest which caused ≥ 90% mortality. In addition, the extract caused rapid blood digestion in ticks as well as obvious decrease in their oviposition (40% oviposition for the highest concentration compared to 100% for the control group), hatchability (40-45% egg hatching across concentrations compared to 78% for the control group) and fertility index (0.32 compared to 0.56 for the control group). As the estimated LC50 and LC95 were recorded 63.84 and 1046.11 mg/mL, respectively, the study also revealed that dipping in 200 mg/mL of C. colocynthis alcoholic extract is sufficient to control H. dromedarii as it recorded >85% acaricide efficacy, so it might be selected to be the effective dose. Moreover, gas chromatography-mass spectrometry (GC-MS) analysis was carried out on alcoholic extract of C. colocynthis fruits and detected its major constituents of decanoic, oleic, linoleic, carbamic and vaccenic acids which have acaricidal activities impacting tick feeding, digestion and survival. The combined effects of alcoholic extract of C. colocynthis fruits indicated by high mortality rate as well as decreased fecundity and reproductive efficacy supports its effectiveness as a promising management agent for the control of H. dromedarii.

Screening of essential oils with acaricidal activity against Haemaphysalis longicornis (Acari: Ixodidae) and analysis of active components.

Haemaphysalis longicornis (Acari: Ixodidae) is an important vector of numerous pathogens and poses a great threat to veterinary and public health. Commercially available tick repellents are extensively used and primarily comprise synthetic molecules; however, there are concerns over their safety and environmental impacts. Biologically based acaricides, particularly the plant-derived essential oils (EOs), may constitute an appealing alternative. We screened 20 different EOs by packet tests of unfed H. longicornis nymphs, and found that EOs of cinnamon, clove and chamomile were the most toxic (mortality > 80 %). Cinnamon EO had the most competitive acaricidal activity, with lethal concentration 50 (LC50) rates of 0.4530 %, 0.2316 % and 0.0342 % (v/v) for unfed adults, nymphs and larvae, respectively. Furthermore, 5.00 % (v/v) cinnamon EO showed reproductive inhibition against H. longicornis, with significantly higher rates of oviposition reduction (53.19 %) and hatching reduction (46.21 %) compared with the negative control group. Composition analysis of cinnamon EO by gas chromatography-mass spectrometry (GC-MS) revealed that the major chemical compounds were trans-cinnamaldehyde (72.21 %) and cinnamic acid (19.45 %), with the former showing similar levels of acaricidal activity and oviposition inhibition as cinnamon EO. This study has demonstrated the potential of cinnamon EO and trans-cinnamaldehyde as natural acaricides against H. longicornis, and is the first to characterize their oviposition inhibition activity.

Efficacy of synergistic activity of seed oils from Carthamus tinctorius (Safflower) and Nasturtium officinale (Watercress) on the lethality of the cattle tick Hyalomma scupense (Acari: Ixodidae).

Background: Ticks and tick-borne diseases are a severe economic and public-health problem for cattle producers. The emergence of acaricide resistance to synthetic chemical treatments has prompted interest in developing alternative tick control methods. Aim: The main objective of the current research was to identify the chemical structure of Carthamus tinctorius and Nasturtium officinale seed oils and to assess their anti-tick properties against Hyalomma scupense ticks both alone and in combination (1:1). Methods: Analytical methods were used to analyze the chemical components. For in vitro assays, adults of H. scupense were immersed in C. tinctorius and N. officinale seed oils at 100, 125, 200, and 300 mg/ml concentrations; for 5 minutes. Larvae of H. scupense were dipped in 25, 50, 100, 125, 200, and 300 mg/ml doses of seed oils; the mortality percentage was determined after 24 hours. Results: The seed oil safflower was mainly composed of linoleic acid (84.48%), followed by palmitic acid (6.54%) and stearic acid (3.77%). Meanwhile, watercress seed oil was mainly composed of linolenic acid (50.78%), gondoic acid (13.57%), linoleic acid (10.58%), palmitic acid (8.02%), and erucidic acid (6.62%). The Adults Immersion Test showed the sensitivity of ticks to C. tinctorius and N. officinale seed oil: C. tinctorius seed oil caused (95%) mortality of H. scupense at 300 mg/ml, while N. officinale seed oil induced (88.68%) mortality at the same concentration. At a 200 mg/ml concentration, C. tinctorius and N. officinale oil combined caused 100% mortality. Tested oils showed larvicidal efficacy. LC50 values for C. tinctorius and N. officinale seed oils were 84.16 and 61.78 mg/ml, respectively, in 24 hours. LC50 value of oils association (50% C. tinctorius: 50% N. officinale) was 47.96 mg/ml.The mixture of seed oils from two plants tested against H. scupense larvae and adult females at a 1:1 ratio showed synergistic interaction. Conclusion: Seed oils tested alone, and the mixture could be used as an alternative solution in the fight against ticks.

Persistence Enhancement of a Promising Tick Repellent, Benzyl Isothiocyanate, by Yeast Microcarriers.

Phenethyl isothiocyanate isolated from Armoracia rusticana root oil and its derivatives were tested at different doses in a bioassay designed to evaluate repellency against individual Haemaphysalis longicornis nymphs. Among the tested compounds, benzyl isothiocyanate exhibited repellency against H. longicornis nymphs at the lowest dose of 0.00625 mg/cm2, followed by phenethyl isothiocyanate (0.0125 mg/cm2) and phenyl isothiocyanate (0.025 mg/cm2). The behavioral responses of H. longicornis nymphs exposed to benzyl isothiocyanate and phenethyl isothiocyanate indicated that the mode of action of these compounds can be mainly attributed to the vapor phase. Encapsulated benzyl isothiocyanate showed repellency up to 120 min post-application at 0.1 mg/cm2, whereas pure benzyl isothiocyanate showed repellency up to 60 min post-application at 0.1 mg/cm2. The present study suggests that benzyl isothiocyanate is a potential repellent for protection against H. longicornis nymphs, and encapsulation in yeast cells may enhance the repellency effect.

Effects of acaricidal essential oils from Lippia sidoides and Lippia gracilis and their main components on vitellogenesis in Rhipicephalus microplus (Canestrini, 1888) (Acari: Ixodidae).

Rhipicephalus microplus is an important cattle tick, and resistant strains to synthetic compounds have been widespread. The combined effects of different essential oil compounds enhance biological activity and reduce selection for the development of target organism resistance. Essential oils of two different genotypes of each of Lippia sidoides and Lippia gracilis and their main components, the isomers thymol and carvacrol, have acted as acaricides against R. microplus. Little is known about the effects of the essential oils of L. sidoides and L. gracilis and thymol and carvacrol on the morphophysiology of R. microplus ovaries. This study aimed to identify the morphological changes in the ovaries of R. microplus females treated with essential oils from two different genotypes of each of L. sidoides (102 and 103) and L. gracilis (106 and 201) and the terpenes thymol and carvacrol through histological techniques. The LC50 and LC75 of essential oils and thymol and carvacrol were used for Adult Immersion Test (AIT) with groups of five fully engorged females of R. microplus. A negative control (DMSO 3% solution) was performed. Seven days after the AIT, the ticks were dissected to collect ovaries and their histologic analysis. Only the group treated with the essential oil of L. gracilis genotype 106 at the LC50 had no change compared with the control. The other groups showed the following changes in oocytes I to V: vacuolation, chorion deformation, disorganization of yolk granules, and irregularities at the cell periphery, causing incomplete process of vitellogenesis. Thus, the essential oils tested in this study may be potent products for the control of cattle ticks and thereby preventing further life cycles.

Acaricidal activity of the hexanic and hydroethanolic extracts of three medicinal plants against southern cattle tick Rhipicephalus (Boophilus) microplus (Acari: Ixodidae).

The southern cattle tick Rhipicephalus microplus is a major problem for the cattle industry in tropical and subtropical areas worldwide. Chemical products are commonly applied to control it; however, their indiscriminate use has resulted in the appearance of resistant lineages. In the last decades, plants have been used as an alternative to conventional acaricidal drugs, as several plant compounds repel activity, decrease the reproductive potential and reduce the survival rate of ticks. For this reason, the in vitro efficacy of hexanic and hydroalcoholic extracts of Randia aculeata, Moringa oleifera and Carica papaya were evaluated against the larvae and engorged females of R. microplus. Larval packet tests and adult immersion tests were performed with seven concentrations of each of the extracts. The extracts obtained with hydroethanolic solution (polar solvent) exhibited a higher acaricidal activity than extracts prepared with n-hexane (non-polar solvent). Hydroethanolic extracts of R. aculeata seed and shell showed the highest larvicidal activity against R. microplus (100 and 91% mortality, respectively) at a concentration of 100 mg/mL. Randia aculeata (seed and shell), M. oleifera and C. papaya treatments at the same concentration (100 mg/mL) also resulted in adult mortality of 85, 75, 66 and 55%, respectively. The adult immersion test showed that hydroethanolic extracts derived from R. aculeata seed significantly reduced the index of egg laying and increased the percentage inhibition of oviposition of female ticks at a concentration of 100 mg/mL. These results indicate that the tested extracts exhibit acaricidal activity and could be considered as potential agents for the development of alternative natural acaricides against R. microplus.

Acaricidal Activity of Colchicum autumnale (autumn crocus) Extract against Hyalomma spp. In vitro.

Hyalomma spp. is responsible for the transmission of protozoan, bacterial, rickettsial, and viral diseases and causes huge economic loss to the livestock industry.Recently, there is a wide number of promising attempts to evaluate and use herbal preparations for ticks control.This study aimed to evaluate the acaricidal activity of aqueous and ethanol extracts of Colchicum autumnale (C. autumnale) rhizome and leaf against the Hyalomma spp. in vitro. The acaricidal activities of the Colchicum leaf aqueous (CLA), Colchicum leaf ethanolic (CLE), Colchicum rhizome aqueous (CRA), and Colchicum rhizome ethanolic (CRE) extracts were evaluated at concentrations of 50, 100, and 150 mg/ml and controls (distilled water and Cypermethrin) following 0.25, 0.5, 0.75, and 1 h of exposure. It is worth mentioning that the spraying method was used in these experiments. Data were analyzed through GraphPad Prism 5 software. In addition, the chemical composition of aqueous leaf extract was analyzed using gas chromatography-mass spectrometry (GC&ndash;MS). The carbamodithioic acid (30.04%) was the major chemical constituent identified. Based on the results, CLA, CLE, CRA, and CRE extracts had an acaricidal effect; however, this effect was more potent in CLE. The CLE extract showed a 100% mortality rate at 50, 100, and 150 mg/ml concentrations and 1 h of exposure. The effectiveness of CRA on the Hyalomma spp. was very low. The median lethal concentration (LC50) values were obtained at 100 mg/ml. The results indicated that C. autumnaleleaves contained potent acaricidal ingredients and might provide new acaricidal compounds for the effective control of Hyalomma spp. However, further studies are required to evaluate the efficacy of C. autumnale in vivo.

Acaricidal activity of Cinnamomum cassia (Chinese cinnamon) against the tick Haemaphysalis longicornis is linked to its content of (E)-cinnamaldehyde.

BACKGROUND: The tick Haemaphysalis longicornis (Neumann) is a well-known vector of numerous pathogens of veterinary and medical importance. Various control strategies, including the use of synthetic pesticides, have been developed to control this tick species. However, demand for effective and safe alternative pesticides is increasing due to the adverse effects associated with the intensive and injudicious use of synthetic pesticides, which include undesirable effects on non-target species and environmental pollution. Hence, the acaricidal activity of the extract and the essential oil of Cinnamomum cassia (Chinese cinnamon) and their major components, and the underlying mechanisms of this activity, were evaluated against unfed larvae and nymphs of H. longicornis. METHODS: The components of the extract and essential oil of C. cassia were determined by gas chromatography-mass spectrometry, and their larvicidal and nymphicidal activity were evaluated using the larval and nymphal packet test. The underlying detoxification mechanism was elucidated by targeting in vivo esterase and monooxygenase activity, and the toxicological effect was assessed on non-target Tenebrio molitor and Harmonia axyridis by topical application in open Petri dishes. RESULTS: (E)-cinnamaldehyde was the predominant component of the extract (50.79%) and essential oil (89.95%). The 50% lethal concentration (LC50) for larvae and nymphs treated with the extract was 11.56 and 49.18 mg/mL, respectively. The essential oil, (E)-cinnamaldehyde and fenvalerate exhibited acaricidal activity, with LC50 values of 3.81, 3.15, and 0.14 mg/mL, respectively, against the larvae, and 21.31, 16.93, and 1.89 mg/mL, respectively, against the nymphs. (E)-cinnamaldehyde significantly increased esterase and monooxygenase activity in both larvae and nymphs. Unlike fenvalerate, C. cassia essential oil and (E)-cinnamaldehyde did not cause mortality of T. molitor or H. axyridis adults. CONCLUSIONS: This study demonstrates that C. cassia essential oil and (E)-cinnamaldehyde have the potential to be developed into botanical-based larvicidal and nymphicidal agents for tick control.

Evaluation on two types of paramyosin vaccines for the control of Haemaphysalis longicornis infestations in rabbits.

BACKGROUND: Haemaphysalis longicornis is an obligate hematophagous ectoparasite that transmits a variety of pathogens causing life-threatening diseases in humans and animals. Paramyosin (Pmy) is not only an invertebrate-specific myofibrillar protein but also an important immunomodulatory protein. Therefore, it is one of the ideal candidate antigens for vaccines. METHODS: We conducted two vaccine trials to evaluate the protective efficacy of Pmy recombinant protein (rPmy) and peptide vaccine (KLH-LEE). Each rabbit was immunized with three doses of rPmy or KLH-LEE adjuvanted with Freund's complete/incomplete at 500 µg/dose at 2-week intervals before challenge with 40 female H. longicornis/rabbit. PBS plus adjuvant, Trx or KLH was used as control group. The antibodies of rabbits were detected by ELISA. Then, female ticks were fed on the rabbits until detachment. RESULTS: ELISA results showed that both vaccines induced rabbits to produce antibodies. Compared with the Trx group, the engorgement weight, oviposition and hatchability of the rPmy group decreased by 8.87%, 26.83% and 38.86%, respectively. On the other hand, engorgement weight, oviposition and hatchability of female ticks in the KLH-LEE group correspondingly resulted in 27.03%, 53.15% and 38.40% reduction compared with that of the KLH group. Considering the cumulative effect of vaccination on the evaluated parameters, results showed 60.37% efficacy of the rPmy vaccine formulation and 70.86% efficacy in the KLH-LEE group. CONCLUSIONS: Pmy and particularly epitope LEE have potential for further development of an effective candidate vaccine to protect the host against tick infection. GRAPHIC ABSTARCT.

Acaricidal activity of Artemisia herba-alba and Melia azedarach oil nanoemulsion against Hyalomma dromedarii and their toxicity on Swiss albino mice.

Biopesticides such as essential oils (EOs) are considered an improvement for integrated pest control as they appear to be less toxic to the environment than chemical acaricides. The current study aimed to evaluate the acaricidal activity of Artemisia herba-alba and Melia azedarach oil loaded nano-emulsion as alternatives for chemical acaricides against the camel tick Hyalomma dromedarii, besides evaluating their toxic effect on Swiss albino mice. Transmission electron microscopy (TEM) and Fourier transform infrared spectroscopy (FTIR) were used for the characterization of loaded nano-emulsions.The immersion test was used for the bioassay of both loaded nanoemulsions on tick stages (egg, nymph, larva, and adult). Mortality percentages and LC50 values of each tick stage were calculated. Reproductive performance for the survived engorged females after treatment was monitored. The toxicity of both loaded nano-emulsions was evaluated on Swiss albino mice by an oral dose of 1500 mg/kg/day for five successive days. The hematological, biochemical, and histopathological changes were evaluated. TEM characterization revealed spherical droplets for A. herba-alba and M. azedarach oil loaded nano-emulsion with droplet size ranging from 62 to 69 nm and 52-91 nm, respectively. FTIR revealed the absence of extra peaks in the loaded nano-emulsions that confirmed no chemical changes existed by ultrasonication. The LC50 values of A. herba-alba and M. azedarach oil loaded nano-emulsion on embryonated eggs, larvae, engorged nymphs, and unfed adults were 0.3 and 1.1%, 0.7 and 1.7%, 0.3 and 0.4%, 4.4 and 22.2%, respectively. The egg productive index (EPI), egg number, and hatchability percentage were lower in the treated females compared with Butox 5% (deltamethrin) and control. The hematological picture and biochemical analysis revealed insignificant changes in the treatment group compared with the negative control group. The liver of the A. herba-alba and M. azedarach oil loaded nano-emulsion treated group exhibited vacuolar degeneration and infiltration of lymphocytic cells. The kidney of mice treated with A. herba-alba and M. azedarach oil loaded nano-emulsion showed hemolysis and slight degeneration of epithelial cells of tubules. It is concluded that A. herba-alba and M. azedarach oil loaded nano-emulsion have good acaricidal activity against camel tick H. dromedarii.