Chemical Acaricides Research Articles

Lethal and sublethal effects of several plant compounds compared to spiromesifen against Tetranychus turkestani

Summary Tetranychus turkestani Ugarov and Nikolski is one of the main agricultural pests of southwestern Iran and some other tropical regions. In the present study, fumigant activity of three essential oils extracted from Rosmarinus officinalis L., Mentha longifolia L. and Eucalyptus globulus Labill., and contact toxicity of two biopesticides (Tondexir and Palizin) on T. turkestani mite females were investigated and compared with that of spiromesifen, a chemical acaricide. Also, sublethal effects of the tested compounds at 50% lethal concentration (LC50) were estimated on the biological parameters of T. turkestani. The LC50 values for E. globulus, R. officinalis and M. longifolia essential oils were 12.50, 11.52 and 4.00 μl/l air and for spiromesifen, Tondexir and Palizin were 10.98, 327.34 and 858.13 ppm, respectively. All tested compounds significantly reduced adult female longevity, equally to the chemical acaricide spiromesifen. Fecundity also decreased in all treatments and this reduction was even higher for plant essential oils than the other compounds. Palizin, E. globulus and M. longifolia significantly reduced the hatchability of T. turkestani eggs similarly to spiromesifen. According to the results, the tested plant compounds are effective against T. turkestani and may be applied as suitable alternatives to synthetic pesticides against this crop pest.

Optimizing long-acting acaricide use for integrated tick management of Rhipicephalus australis-infesting cattle in New Caledonia.

The tick Rhipicephalus australis, formerly known as Rhipicephalus microplus, is the most economically important ectoparasite of livestock in New Caledonia, affecting cattle health and production. Decades of control attempts based on the application of chemical acaricides have exerted a strong selective pressure on R. australis populations, some of which have evolved resistance to these treatments. Research to develop integrated tick control programs is now focused on decreasing applications of chemicals. This study reports the implementation of a method of pasture and herd management involving minimal strategic use of long-acting acaricides, here defined as those having substantial efficacy against larvae for several weeks. Diverse parameters concerning the utilization of long-acting acaricides in association with pasture and herd management on 21 New Caledonian farms over a 5-year period were analyzed to optimize their strategic use. Longer larval acaricidal effect was achieved with a commercial pour-on formulation of fluazuron than with a commercial injectable (subcutaneous) formulation containing 3.15% ivermectin. Pasture and herd management allowed an increase in the delay between a long-lasting acaricide application and the subsequent acaricide treatment from 11.0weeks to 17.7weeks. However, if ticks were detected and reported by producers on the day of a long-acting acaricide application, the delay to the following treatment was reduced from 18.5weeks to 11.2weeks. The impact of a long-acting acaricide treatment on larval populations in pastures was greatest with a stocking rate of 5 animals per hectare grazing during 1week. These results provide science-based evidence to cattle producers for adaptive integrated tick management in order to delay the development of acaricide resistance.

Field Experiment Effect on Citrus Spider Mite Panonychus citri of Venom from Jellyfish Nemopilema nomurai: The Potential Use of Jellyfish in Agriculture

Jellyfish are rich in resources and widely distributed along coastal areas. As a potential approach to respond to jellyfish blooms, the use of jellyfish-derived products is increasing. The citrus spider mite (Panonychus citri) is one of the key citrus pests, negatively impacting the quality and quantity of oranges. Due to the resistance and residue of chemical acaricides, it is important to seek natural substitutes that are environmentally friendly. The field efficacy of the venom from the jellyfish Nemopilema nomurai against P. citri was assayed in a citrus garden. The frozen N. nomurai tentacles were sonicated in different buffers to isolate the venom. The venom isolated by PBS buffer (10 mM, pH 6.0) had the strongest acaricidal activity of the four samples, and the corrected field efficacy 7 days after treatment was up to 95.21%. This study demonstrated that jellyfish has potential use in agriculture.

Tick treatment practices in the field: Access to, knowledge about, and on-farm use of acaricides in Laikipia, Kenya

The prevention of tick-borne diseases is a major challenge for livestock production globally. Tick control strategies include the use of acaricides, but the prescribed strategies do not achieve the desired results in several countries, including Kenya. To better understand how tick treatment practices, contribute to reported tick treatment failures, we assessed livestock owners’ acaricide procurement, level of knowledge about acaricides and tick resistance, and how they apply acaricides. We also assessed the quality of the commonly available acaricides. We focused on three livestock systems in Laikipia County, Kenya: two private ranches; one community ranch whose members communally graze their cattle and acquire and apply acaricides; and individual livestock owners in two pastoral communities who individually graze their cattle and acquire and apply acaricides. Through interviews and focus group discussions we assessed; access to acaricides, livestock owners’ knowledge, and acaricide use practices; interview data were triangulated with participant observations (n = 107). We analysed nine commonly used acaricides to determine the active ingredient concentration and we determined the concentration of active ingredients in acaricide dilutions collected on farms. All livestock owners had access to and used chemical acaricides for tick control, predominantly amitraz-based. Private ranchers bought one amitraz-based acaricide in bulk directly from the manufacturer, while all other livestock owners bought from agrovet shops. The livestock owners acquired knowledge about acaricides from their own experiences and through experience-based recommendations from peers, but not from the technical information provided by the manufacturers and agrovet shops. All pastoral livestock frequently changed acaricide brand and active ingredient class. A large majority of pastoralists (86%) mixed acaricide brands within and across active ingredient classes; a smaller majority (56%) mixed acaricides with crop pesticides and insecticides. Our lab tests confirmed the content description on the labels bought from agrovet shops. However, on-farm acaricide dilutions from all three livestock systems deviated from the level recommended for effective treatment. If too diluted, the acaricide does not kill ticks, promoting resistance development. If too concentrated, this increases environmental contamination and raises public health concerns. Livestock owners lack a technical understanding of the functioning of acaricides, compromising their use and effectiveness. The widely adopted mixing of acaricides with insecticides and pesticides raises serious health concerns.

Interactive Gene Expression Between Metarhizium anisopliae JEF-290 and Longhorned Tick Haemaphysalis longicornis at Early Stage of Infection.

The longhorned tick, Haemaphysalis longicornis (Acari: Ixodidae), is a hard tick and a vector for severe fever with thrombocytopenia syndrome (SFTS) virus. The number of patients infected with SFTS is rapidly increasing. Recently, the invertebrate pathogen Metarhizium anisopliae JEF-290 was reported to be useful to control the tick as an alternative to chemical acaricides, which are not easily applicable in human living areas where the tick is widely spread. In this study, we analyzed how the tick and the fungal pathogen interact at the transcriptional level. Field-collected tick nymphs were treated with JEF-290 conidia at 1 × 108 conidia/ml. In the early stage of infection with 2.5% mortality, the infected ticks were subjected to RNA sequencing, and non-infected ticks and fungal masses served as controls. Fungus and tick genes were mostly up-regulated at the early stage of infection. In the gene set enrichment analysis of the infecting fungus, catabolic processes that included lipids, phospholipids, and detoxification processes, the response to oxidative stress, and toxic substances were significantly up-regulated. In this fungal up-regulation, various lipase, antioxidant enzyme, and hydrolase genes were highly transcribed. The gene set enrichment analysis of the infected tick showed that many peptide synthesis processes including translation, peptide metabolism, ribonucleotide metabolism, and energy production processes that included ATP generation and ADP metabolism were significantly up-regulated. Structurally, mitochondria and ribosome subunit genes in ticks were highly transcribed to upregulate these processes. Together these results indicate that JEF-290 initiates process that infects the tick while the tick actively defends against the fungal attack. This work provides background to improve our understanding of the early stage of fungal infection in longhorned tick.

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 1500mg/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 69nm and 52-91nm, 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.

Breeder’s knowledge on ticks and tick-borne diseases and management strategies in Menoua division (Western region of Cameroon)

Cattle are challenged by numerous tick species in the western highlands of Cameroon. Consequently, several tick-borne diseases (TBDs) are known to be endemic in this region. To face this situation, farmers mainly resort to the use of chemical acaricides. But to date, no study has focused its attention on the evaluation of tick control methods in this area. Therefore, the present study was designed to assess the knowledge and practices of breeders against ticks and TBDs in Menoua Division. A descriptive cross-sectional survey was conducted in 4 Sub-divisions of Menoua Division. Breeders who consented to the study were interviewed using semi structured questionnaires. Interviews were conducted in French, English or local language (Fulfulde). The questionnaire was made of 25 questions sectioned into; (i) farmer’s knowledge on ticks and TBDs, (ii) treatment practices of infested animals and, (iii) the estimated amount spent for vector control of ticks. All the breeders interviewed had a good knowledge on ticks and the main direct effects they inflict on animals. However, only 7% of respondents associated ticks to the main TBDs. Manual removal and chemical control using synthetic acaricides (usually pyrethrinoids) were the main methods of tick control. Despite the multiplication of treatments (on average 1/week), their effectiveness was not perceived by almost all breeders (82.0%; n = 82) who complained of treatment failures, regardless of the sites and molecules used. In addition, each farm spent on average 8, 521 ± 2, 968 CFAF (US$16.08 ± 5.6)/month for vector control. That is a cumulative amount of CFAF 588, 000 (US$1, 109.45)/month and CFAF 7, 056, 000 FCFA (US $ 13, 313.49)/year for all the farms visited. The lack of pasture (OR = 43.5; p

Entomopathogenic Fungi for Tick Control in Cattle Livestock From Mexico.

Ticks are one of the main economic threats to the cattle industry worldwide affecting productivity, health and welfare. The need for alternative methods to control tick populations is prompted by the high prevalence of multiresistant tick strains to the main chemical acaricides and their ecological consequences. Biological control using entomopathogenic fungi (EPF) is one of the most promising alternative options. The objective of this paper is to review the use of EPF as an alternative control method against cattle ticks in Mexico. Metarhizium anisopliae sensu lato (s.l.) and Beauveria bassiana s.l. are the most studied EPF for the biological control of ticks in the laboratory and in the field, mainly against Rhipicephalus microplus; however, evaluations against other important cattle ticks such as Amblyomma mixtum and R. annulatus, are needed. A transdisciplinary approach is required to incorporate different types of tools, such as genomics, transcriptomics and proteomics in order to better understand the pathogenicity/virulence mechanism in EPF against ticks. Laboratory tests have demonstrated the EPF efficacy to control susceptible and resistant/multiresistant tick populations; whereas, field tests have shown satisfactory control efficiency of M. anisopliae s.l. against different stages of R. microplus when applied both on pasture and on cattle. Epidemiological aspects of ticks and environmental factors are considered as components that influence the acaricidal behavior of the EPF. Finally, considering all these aspects, some recommendations are proposed for the use of EPF in integrated control schemes for livestock ticks.

EVALUATING SIDE EFFECTS OF THREE PESTICIDES AND FIVE HEAVY METALS ON PREDATORY MITES IN IPM PROGRAMS

Integrated Pest Management (IPM) is a crucial point in awareness of pesticides and heavy metal’s side effects on non-target beneficial arthropods. Studies on the toxicity of selected pesticides and heavy metals to predatory mite species: Phytoseiulus persimilis and Amblyseius swirskii (Athias-Henriot) have been carried out. insecticides were used: (greeno 10% Hexathiazox), Agri-Flex® 18.56% (abamectin,Thiamethoxam) and Baylor - 24 % (Chlorfenapyr) beside many of concentrations of heavy metals (As, Cu, Pb, Mn, and Zn). Acalypha wilkesiana (Müll. Arg). The results showed that the highest effect of chemical acaricides on predators Baylor - 24% with a percentage of 90.6%, and the lowest mortality was Agri-Flex® 18.56% with a percentage of 47.5% in dose 2ml double of the recommended dose. Also, in dose (0.25,0.5 and 1ml), the highest toxicity was Baylor - 24 % in secondlevel greeno 10% finally was Agri-Flex® 18.56%. In P. persimilis, the effective dose 0.25 ml with a mortality rate of 7.5%. Also, results showed that the highest mortality rate against A. swirskii were Baylor - 24 %in concentration 2 ml was 90.6 % mortality, and lowest mortality rate in the concentration of .025 ml was 11.30 then greeno 10% the highest mortality was 44%, and the lowest conc in .025 ml was 5%.results showed that the highest or most toxic heavy metals of death rate on P. persimilis also M. Swerskii in Cd treatment, and the lowest mortality was Zn treatments among Mortality rate and repellent rate

Contact toxicity of a new acaricide, SYP-9625, to the natural predator, Chrysopa pallens

• We first evaluated the contact toxicity of an acaricide, SYP-9625, on C. pallens . • SYP-9625 only had mild detrimental impacts on insect growth and metamorphosis. • SYP-9625 had significant adverse effects on adult survival and Vg expression. • The paper will strengthen the compatibility of acaricide and natural predators. SYP-9625 is a novel acaricide synthesized by using cyenopyrafen as lead compound. It is characterized by high acaricidal activity and low toxicity to mammals and has been widely used in Chinese farming areas. However, to date, the toxicity of SYP-9625 to natural enemy insects remained unclear. Here, we evaluated the effects of SYP-9625 on survival, growth, metamorphosis and reproduction of a common natural predator, Chrysopa pallens . Contact toxicity bioassay revealed that SYP-9625 had no significant detrimental impacts on egg hatchability, larval survival and duration, pupation rate, pupal mass, emergence rate, digestive enzyme activity, reproductive output and offspring quality, but drastically reduced adult survival rate, adult lifespan, secondary follicle size and vitellogenin (Vg) transcripts by 27.8%, 15.4%, 33.4% and 59.4%, respectively, suggesting a low contact toxicity of SYP-9625 to C. pallens . This study will strengthen the compatibility of chemical acaricide and natural enemy insects in integrated pest management.

Innovative granular formulation of Metarhizium robertsii microsclerotia and blastospores for cattle tick control

The tick Rhipicephalus microplus poses a serious threat to the cattle industry, resulting in economic losses aggravated by tick resistance to chemical acaricides. Strains of Metarhizium spp., a well-known group of entomopathogenic fungi, can contribute to managing this ectoparasite. We explored two novel granular, microsclerotia- or blastospores-based formulations of Metarhizium robertsii for R. microplus control under semi-field conditions. Fungal persistence in soil was also observed for 336 days. The experiment used pots of Urochloa decumbens cv. Basilisk grass, treated with 0.25 or 0.5 mg of granular formulation/cm2 (25 or 50 kg/ha) applied to the soil surface prior to transferring engorged tick females onto the treated soil. The fungal granules yielded more conidia with subsequent sporulation under controlled indoor conditions than in the outdoor environment, where the levels of fungus rapidly declined over time. Metarhizium-root colonization ranged from 25 to 66.7% depending on the propagule and rate. Fungal formulations significantly reduced the number of tick larvae during the humid season, reaching at least 64.8% relative efficacy. Microsclerotia or blastospores-granular formulations of M. robertsii can reduce the impact of R. microplus, and thus prove to be a promising tool in the control of ticks.

Homology modeling, docking, molecular dynamics and in vitro studies to identify Rhipicephalus microplus acetylcholinesterase inhibitors

Rhipicephalus microplus is an important ectoparasite of cattle, causing considerable economical losses. Resistance to chemical acaricides has stimulated the search for new antiparasitic drugs, including natural products as an eco-friendly alternative of control. Flavonoids represent a class of natural compounds with many biological activities, such as enzyme inhibitors. Acetylcholinesterase is an essential enzyme for tick survival that stands out as an important target for the development of acaricides. This work aimed to predict this 3D structure by homology modeling and use the model to identify compound with inhibitory activity. The model of R. microplus AChE1 (RmAChE1) was constructed using MODELLER program. The optimization and molecular dynamic investigation were performed in GROMACS program. The model developed was used, by molecular docking, to evaluate the anticholinesterase activity of flavonoids (quercetin, rutin, diosmin, naringin and hesperidin) and an acaricide synthetic (eserine). Additionally, in vitro inhibition of AChE and larval immersion tests were performed. The model of RmAChE1 showed to be sterically and energetically acceptable. In molecular dynamics simulations, the 3D structure remains stable with Root Mean Square Deviation = 3.58 Å and Root Mean Square Fluctuation = 1.43 Å. In molecular docking analyses, only eserine and quercetin show affinity energy to the RmAChE (Gridscore: −52.17 and −39.44 kcal/mol, respectively). Among the flavonoids, quercetin exhibited the best in vitro inhibition of AChE activity (15.8%) and mortality of larvae tick (30.2%). The use of in silico and in vitro techniques has shown that quercetin showed promising anti-tick activity and structural requirements to interact with RmAChE1. Communicated by Ramaswamy H. Sarma

Determination of resistance status to amitraz in the cattle tick Rhipicephalus (Boophilus) microplus from Luzon, Philippines, through bioassay and molecular analysis.

Tick infestations and tick-borne diseases are among the leading causes of economic losses in the cattle industry worldwide. Amitraz is one of the most commonly used chemical acaricides against the cattle tick Rhipicephalus (Boophilus) microplus. Resistance to amitraz has been reported in many countries but not in the Philippines. This study aimed to determine whether cattle ticks from provinces in northern and southern Luzon, Philippines, are resistant to amitraz. Engorged or nearly engorged female ticks were collected from 21 farms and allowed to lay eggs to produce larvae. Larval packet test (LPT) was performed using three concentrations of amitraz, as well as a negative control (diluent). Reverse-transcription polymerase chain reaction (RT-PCR) was also performed to amplify a fragment of the octopamine/tyramine receptor gene and was subjected to sequence analysis by multiple nucleotide and amino acid sequence alignments together with reference strain, amitraz-susceptible strain, and amitraz-resistant strain reported in other countries. LPT results suggest the absence of resistance in all of the tested populations. Interestingly, analysis of the octopamine/tyramine receptor amino acid sequence revealed four out of 14 larval pools having substitutions similar to that of the reported amitraz-resistant strains. Although no apparent resistance was observed in this study, prudent use of amitraz should be practiced as the development of resistance is still likely to occur in the future.

Gene cloning, analysis and effect of a new lipocalin homologue from Haemaphysalis longicornis as a protective antigen for an anti-tick vaccine

Haemaphysalis longicornis is distributed worldwide and transmits a variety of pathogens, causing human and animal disease. Use of chemical acaricides, as a primary tick control method, has several disadvantages, including acaricide resistance, environmental damage and residue accumulation in livestock. Development of a livestock vaccination aimed at a tick protective antigen could be an effective, labor-saving and environmentally-friendly method of reducing tick infestation and transmission of tick-borne pathogens. Lipocalins are low molecular weight proteins that play important roles in blood feeding, immune response and reproduction in ticks. In our study, the open reading frame (ORF) of a lipocalin homologue from H. longicornis (HlLIP) was successfully cloned, which consisted of 387 bp encoding a protein of 128 amino acids. The HlLIP protein sequence showed a close sequence homology with Ixodes persulcatus lipocalin. The HlLIP gene was constitutively detected in all developmental stages and in all tissues of the unfed female tick. The ORF of the HlLIP gene was sub-cloned into pET-32a (+) to obtain the recombinant protein (rHlLIP) and its immunogenicity was comfirmed by western blot. A vaccination trial on rabbits against H. longicornis infestation demonstrated that the rHlLIP protein could significantly prolong the period of tick blood feeding, and reduce tick engorged weight, oviposition and egg hatching rate. The vaccination efficacy of the rHlLIP protein was 60.17 % based on engorged weight, oviposition and egg hatching rate of ticks. The results obtained in this study demonstrate that rHlLIP protein is a promising antigen that could potentially be developed as a vaccine against H. longicornis infestation.

POTENSI BIOAKARISIDA EKSTRAK DAUN MIMBA (Azadirachta Indica) DAN BUNGA CENGKEH (Syzygium Aromaticum) TERHADAP TUNGAU PENYEBAB PENYAKIT KREPES PADA JAMUR KUPING

Wood ear mushrooms is commonly known as one of the cultivated edible mushrooms in Indonesia. The demand for this commodity is still steadily high and leads mushroom farmers to optimize its cultivation. For mushroom farmers in Yogyakarta, the prevalence of krepes disease which is caused by a group of species belongs to Acarina order is still high and leads to total loss of harvest. In order to diminish these Acarina species, the use of chemical acaricides is common because of their effectiveness in eradicating the pests despite their toxic effect to the environment. A strategy to provide environmental friendly acaricides which are extracted from plants is therefore important. This research aimed to study the potency of neem leaves (Azadirachta indica) and clove flowers (Eugenia caryophyllata) extracts as bioacarisides for Acarina species as the causal agent of krepes disease. Neem leaves and cloves flowers were extracted using maceration and soxhlet extraction methods, using 96% methanol as their solvents. Obtained crude extracts would then be subjected to phytochemical analysis using qualitative biochemical tests and GC-MS. The effectivity of extracts as bioacaricides were tested both in in vitro and field scale. In vitro tests showed that neem leaves extract and clove flower extract had LC50 value ​​of 0.22% and 0.48% respectively. Field scale test on mushroom baglogs showed that the efficacy of the neem leaves and clove flower to eradicate acarina were above 50%. Taken together, neem leaves and clove flower extracts are potential to be used as bioacaricides in mushroom farming.

Acaricidal Properties of Herbal Extracts against Deltamethrin Resistant Multi-host Cattle Tick Hyalomma anatolicum

Background: Hyalomma ticks are the major ticks prevalent in Gujarat state which are mainly controlled with chemical acaricides. The control strategies are complicated by emergence of resistance against chemical acaricides. Development of resistance against these acaricides has compelled scientists to explore the acaricidal properties of constituents other than chemicals. Methods: Resistance status of Hyalomma anatolicum collected from Gujarat (India) against deltamethrin was assessed by larval packet test, which revealed level I resistance against deltamethrin. The crude ethanolic extracts of leaves of Cymbopogon citratus, Citrus aurantiifolia, Annona squamosa, Carica papaya and Catharanthus roseus were assessed for their acaricidal activity against larvae of deltamethrin resistant H. anatolicum. The efficacy was assessed by measuring per cent larval mortality and determination of LC50 values. Result: Various ethanolic extracts emanated a concentration dependent increase in larval tick mortality. The highest mortality (93.01 ± 0.35%) was observed with the 100mg/ml concentration of ethanolic extract of leaves of C. aurantiifolia. Overall, C. aurantifolia, A. squamosa and C. citratus were found to have significant acaricidal properties with LC95 values of 1.07x102 mg/ml, 6.09x102 mg/ml and 3.62x103 mg/ml respectively. Results of the present study indicated that these plant extracts have potential to be developed as herbal acaricides.

Effect of polyphenols extracted from Punica granatum and Acacia saligna plants on glutathione S-transferase of the cattle tick Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae).

Ticks are hematophageal ectoparasites that transport major pathogens around the world. Glutathione S-transferases (GST) are involved in resistance to acaricide and redox balancing during the life cycle of the tick. The inhibition of tick GST enzymes by certain phenolic compounds, such as phenolic acids and tannins, can be a promising approach to tick control. The objective of this study was to evaluate the effects of Punica granatum red peel and Acacia saligna leaf extracts on Rhipicephalus (Boophilus) annulatus GST activity in order to reduce the resistance of cattle to acaricide. The results showed that P. granatum ethanol extract (70%) contained the highest total phenol content (350 ± 1.2μM GAE g-1), the highest condensed tannin content (270 ± 1.3μM CE g-1) and the highest hydrolysable tannin content (70 ± 5.0μM TAE g-1). Adult immersion test with a dosage of 100mg ml-1 of A. saligna ethanol extracts had a significant mortality of 50% and 75% after 24h and 96h, respectively (p < 0.01). A simple and reproducible procedure was established to purify the whole R. annulatus GST (wRaGST) while a full-length cDNA of GST was cloned from a cDNA library of the local Egyptian cattle tick R. (B.) annulatus (rRaGST). Aqueous extracts of P. granatum inhibited both wRaGST and rRaGST with values of IC50 = 0.114 and 0.07µgml-1, respectively, compared to A. saligna extracts (IC50 values = 2.08 and 1.35µgml, respectively). These inhibitory effects were attributed to the presence of a high tannin concentration (≥ 80%). HPLC analysis indicated the presence of gallic acid and catechin in both extracts, in addition to the rutin, which was only observed in A. saligna extracts. The addition of a tannin inhibitor, polyethylene glycol, suggested the existence of other phenolic compounds in combination with catechins responsible for inhibiting the activity of these extracts. Non-competitive behaviour of catechins may be helpful in preventing, or at least delaying, the development of chemical acaricide resistancein R. annulatus.

Chemical composition and acaricidal activity of Eucalyptus globulus essential oil against the vector of tropical bovine piroplasmosis, Rhipicephalus (Boophilus) annulatus.

Ticks are of great economic importance to humans and animals due to their role in disease transmission. The application of synthetic, chemical acaricides on the animal and/or the environment (the most used tick control method globally) has led to the selection of tick populations that are resistant. Their adverse effects on ecology andhuman and animal health cannot be overemphasised. As a result, the search for alternatives that are natural and can overcome these adverse effects are strongly indicated. Using gas chromatography-mass spectrometry (GC-MS) and adult immersion test (AIT), this study evaluated the chemical composition and acaricidal activity, respectively, of Eucalyptus globulus essential oil (EO) on Rhipicephalus (Boophilus) annulatus ticks. This is a major tick species implicated for the transmission of bovine piroplasmosis in Nigeria. The acaricidal activity was evaluated using different concentrations (0.625, 1.25, 2.5, 5 and 10%) of E. globulus EO. Amitraz (1 and 2%) and cypermethrin (2%) served as the positive control and 2% dimethylsulfoxide in distilled water was the negative control. Three replicates of 10 engorged female ticks each were immersed in the test samples for 2min and the experiment was done twice. The GC-MS analysis identified the major constituents of E. globulus EO as eucalyptol (1,8-cineole) (78%), menthol (20%) and menthone (3%). Eucalyptus globulus EO caused 97% acaricidal mortality at 10% concentration. The lower concentrations reduced tick fecundity up to 90% in a dose-dependent manner. This study provides support for plant EOs as alternative tick control strategy for humans and animals.

Immunoprotection evaluation of the recombinant N-terminal domain of Cys-loop receptors against Rhipicephalus (Boophilus) microplus tick infestation

Rhipicephalus (Boophilus) microplus ticks are obligatory hematophagous ectoparasites of cattle and act as vectors for disease-causing microorganisms. Conventional tick control is based on the application of chemical acaricides; however, their uncontrolled use has increased resistant tick populations, as well as food and environmental contamination. Alternative immunological tick control has shown to be partially effective. Therefore, there is a need to characterize novel antigens in order to improve immunological protection. The aim of this work was to evaluate Cys-loop receptors as vaccine candidates. N-terminal domains of a glutamate receptor and of a glycine-like receptor were recombinantly produced in Escherichia coli. Groups of BALB/c mice were independently immunized with four doses of each recombinant protein emulsified with Freund’s adjuvant. Both vaccine candidates were immunogenic in mice as demonstrated by western blot analysis. Next, recombinant proteins were independently formulated with the adjuvant Montanide ISA 50 V2 and evaluated in cattle infested with Rhipicephalus microplus tick larvae. Groups of three European crossbred calves were immunized with three doses of each adjuvanted protein. ELISA test was used to evaluate the IgG immune response elicited against the recombinant proteins. Results showed that vaccine candidates generated a moderate humoral response on vaccinated cattle. Vaccination significantly affected the number of engorged adult female ticks, having no significant effects on tick weight, egg weight and egg fertility values. Vaccine efficacies of 33% and 25% were calculated for the glutamate receptor and the glycine-like receptor, respectively.

In vitro Analysis of the Efficacy of Selected Commercial Acaricides on The Cattle Tick Rhipicephalus (Boophilus) annulatus (Acari: Ixodidae)

Ticks are important to the Nigerian livestock industry because of their ability to cause huge economic loss directly or indirectly. Chemical acaricides have played important roles in control of tick infestation for many years but development of acaricide resistance as being reported worldwide is a huge problem facing the livestock industry and the veterinarians. Thus, this study evaluated the efficacy of available commercial acaricides on Rhipicephalus (Boophilus) annulatus. Using Adult Immersion Test (AIT): the LC50, inhibition of oviposition, control and resistance rates, gradation of tarsal reflexes were determined for ticks treated with different acaricides at five different concentrations the discriminating dose (DD) (the manufacturer’s recommended dose) and four other different concentrations of two below and two above the DD with the aid of prohibit regression. The association between weight and number of eggs laid was analyzed by linear correlation. Macrocyclic lactone(66.7%), Phenylpyrazole (62.5%), Cypermethrin (62.5%), Formamidine(33.3%) and Phenylpyrazole(13% egg production), Macrocyclic lactone (18% egg production), Cypermethrin(20% egg production), Formamidine(49% egg production) have the highest to lowest percentage control and oviposition inhibition values respectively. The LC50 of (Formamidine), (Phenylpyrazole), (Synthetic pyrethroids), (Macrocyclic lactone) against adult R. (B.) annulatus were 100, 8000, 2300 and 6000ppm respectively.The demonstration of acaricide resistancedevelopmentas demonstrated by the ticks in laying of eggs after dipping in acaricideis a call to the need for discovery and use of alternative tick control methods with good therapeutic dose and the need for continuous evaluation of available acaricide.