Development of a sensitive electrochemical method to determine amitraz based on perylene tetracarboxylic acid/mesoporous carbon/Nafion@SPCEs.

A cutting-edge electrochemical method is presented for precise quantification of amitraz (AMZ), a commonly used acaricide in veterinary medicine and agriculture. Leveraging a lab-made screen-printed carbon electrode modified with a synergistic blend of perylene tetracarboxylic acid (PTCA), mesoporous carbon (MC), and Nafion, the sensor's sensitivity was significantly improved. Fine-tuning of PTCA, MC, and Nafion ratios, alongside optimization of the pH of the supporting electrolyte and accumulation time, resulted in remarkable sensitivity enhancements. The sensor exhibited a linear response within the concentration range 0.01 to 0.70 µg mL-1, boasting an exceptionally low limit of detection of 0.002 µg mL-1 and a limit of quantification of 0.10 µg mL-1, surpassing maximum residue levels permitted in honey, tomato, and longan samples. Validation with real samples demonstrated high recoveries ranging from 80.8 to 104.8%, with a relative standard deviation below 10%, affirming the method's robustness and precision. The modified PTCA/MC/Nafion@SPCE-based electrochemical sensor not only offers superior sensitivity but also simplicity and cost-effectiveness, making it a pivotal tool for accurate AMZ detection in food samples. Furthermore, beyond the scope of this study, the sensor presents promising prospects for wider application across various electrochemical analytical fields, thereby significantly contributing to food safety and advancing agricultural practices.

Rapid and sensitive quantitation of amitraz in orange, tomato, and eggplant samples using immunochromatographic assay.

Amitraz (AMT) is a broad-spectrum formamidine insecticide and acaricide. In this study, we produced an anti-AMT monoclonal antibody (mAb) with high performance. The half-maximal inhibitory concentration of the anti-AMT mAb was 4.418 ng/mL, the cross reactivity with other insecticides was negligible, and an affinity constant was 2.06 × 109 mmol/L. Additionally, we developed an immunochromatographic assay for the rapid detection of AMT residues in oranges, tomatoes, and eggplants. The cut-off values were 2000 µg/kg in oranges and tomato samples and 1000 µg/kg in eggplant samples and the calculated limits of detection were 14.521 µg/kg, 6.281 µg/kg, and 3.518 µg/kg in oranges, tomatoes, and eggplants, respectively, meeting the detection requirements for AMT in fruits and vegetables. The recovery rates ranged between 95.8 % and 105.2 %, consistent with the recovery rates obtained via LC-MS/MS. Our developed immunochromatographic assay can effectively, accurately, and rapidly determine AMT residues in oranges, tomatoes, and eggplants.

Competitive displacement and acaricide resistance of two Rhipicephalus (Boophilus) species collected on commercial farms in South Africa.

Rhipicephalus (Boophilus) microplus, an invasive species to Africa, and the endemic R. (B.) decoloratus are of high economic importance in the cattle industry. Invasion of the alien species in South Africa has mostly been reported for traditional communal grazing areas where it seemed to be rapid and, in some cases, even replaced the native species. The alien species is also assumed to already be resistant to acaricides upon invasion. The presence of R. (B.) microplus on commercial farms was therefore investigated and resistance screening of both species to field concentrations of cypermethrin, amitraz, and chlorfenvinphos was determined by means of the larval immersion test. Results showed that only 3.7% (of 383) tick collections submitted were R. (B.) microplus populations. A further 1.6% (of 383) showed co-existence of the two species. Comparing the level of resistance to the acaricides between the two species indicated a mean phenotypic resistance of 66.2 and 26.5% of R. (B.) decoloratus populations to cypermethrin and amitraz, respectively. This was significantly lower for R. (B.) microplus, with 23.0 and 4.1% of its populations resistant to cypermethrin and amitraz, respectively. Closed commercial farming areas seemed to have a preventative advantage for the invasion of R. (B.) microplus and displacement of R. (B.) decoloratus by R. (B.) microplus. Regular monitoring of these two species may be of high importance to prevent unnecessary financial losses due to insufficient control and increased awareness of the threat of Asiatic babesiosis vectored by R. (B.) microplus.

Susceptibility of Amblyomma sculptum, Vector of Rickettsia rickettsii, Ticks from a National Park and an Experimental Farm to Different Synthetic Acaricides.

Amblyomma sculptum is a relevant tick species from a One Health perspective, playing an important role as a vector of Rickettsia rickettsii, the main agent of spotted fever rickettsiosis in Brazil. In this study, we evaluated the susceptibility of two A. sculptum populations from Goiás state (midwestern Brazil) to different acaricides. The first tick population (GYN strain) originated from an experimental farm, where the ticks are annually exposed to acaricides. The second (PNE strain) was collected in a national park (Emas National Park), where the ticks had not been exposed to acaricides. Immersion tests were conducted with 21-day-old laboratory-reared larvae and nymphs originating from adult ticks collected in the areas mentioned above. The chosen acaricides were two synthetic pyrethroids (cypermethrin and deltamethrin), one organophosphate (chlorfenvinphos), one formamidine (amitraz), and two combinations of pyrethroids and organophosphates (cypermethrin, chlorpyrifos and citronellal; cypermethrin, fenthion and chlorpyrifos). Mortality data were used to determine the lethal concentration (LC) values at which 50%, 90%, and 99% of the ticks died (LC50, LC90, and LC99, respectively), and resistance ratios (RR) were calculated based on the LC values. The RR revealed differences between the acaricide-exposed (GYN) and unexposed (PNE) tick strains. The PNE strain larvae and nymphs were susceptible to all the tested acaricides. The GYN strain larvae were tolerant to cypermethrin, whereas the nymphs were tolerant to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal (2 < RR ≤ 10). The GYN strain nymphs were resistant to amitraz (RR > 10). This is the first report of A. sculptum nymphs with resistance to amitraz and tolerance to deltamethrin, chlorfenvinphos, and the combination of cypermethrin, chlorpyrifos, and citronellal.

Study of pattern and outcome of acute poisoning cases at tertiary care hospital in North India.

Introduction: The WHO 2021 data estimate that 2 million lives and 53 million disability-adjusted life-years were lost in 2019 due to exposures to selected chemicals. It is important to know the pattern and outcome of acute poisoning cases for proper planning, prevention and management. Knowing the pattern will also help in designing training modules for primary care physicians to make them aware about newer poisons and their management. Awareness regarding newer poisons consumed is necessary for early identification, initial management and timely referral to higher centres by primary care physicians. This study was performed to see the pattern and outcome of acute poisoning cases in North Indian population and various factors related to outcome. Materials and Methods: This study was conducted in department of medicine of a teaching institute in North India after approval by the Institutional Review Board. Patients admitted in the department during the study and fulfilling the inclusion criteria were enrolled in the study after obtaining consent. Results: A total of 417 patients with poisoning were recruited in the study. Out of 417 patients, majority were males (59.5%). Maximum number of patients were in the age group of 21-30 years (33.8%), and rural population (79.9%) was found to be more affected. Most of the patients were students and private employees. Most common types of poisoning were snakebite (n = 109, 26.1%), organophosphate (n = 49, 11.8%) and aluminium phosphide (n = 39, 9.3%). Out of 417 patients, 349 (83.69%) improved, while 68 (16.3%) expired. Requirement of ventilatory support was most commonly associated with aluminium phosphide poisoning (30.12%) followed by organophosphate poisoning (24.1%). Conclusion: Poisoning was more common in young males and more prevalent in rural population. Pesticides and snakebite were major causes of poisoning. Among suicidal cases, family conflict (problem/altercation with family members/marital discord) was main reason for the consumption of poison. There is need for creation of poison information centre along with separate toxicological units in tertiary care hospitals.

Corrigendum: A clash on the Toll pathway: competitive action between pesticides and zymosan A on components of innate immunity in Apis mellifera.

[This corrects the article DOI: 10.3389/fimmu.2023.1247582.].

A clash on the Toll pathway: competitive action between pesticides and zymosan A on components of innate immunity in Apis mellifera.

Background: The immune system of honeybees includes multiple pathways that may be affected by pesticide exposure decreasing the immune competencies of bees and increasing their susceptibility to diseases like the fungal Nosema spp. infection, which is detected in collapsed colonies. Methods: To better understand the effect of the co-presence of multiple pesticides that interact with bees like imidacloprid and amitraz, we evaluated the expression of immune-related genes in honeybee hemocytes. Results: Imidacloprid, amitraz, and the immune activator, zymosan A, mainly affect the gene expression in the Toll pathway. Discussion: Imidacloprid, amitraz, and zymosan A have a synergistic or an antagonistic relationship on gene expression depending on the level of immune signaling. The presence of multiple risk factors like pesticides and pathogens requires the assessment of their complex interaction, which has differential effects on the innate immunity of honeybees as seen in this study.

In vitro toxicity of combination of amitraz and carvacrol on Demodex canis.

Background: Canine generalized demodicosis is an inflammatory skin disease, which usually has time-consuming and frustrating treatments. Aim: The present study aimed to evaluate the acaricidal activity of carvacrol as a green drug and a combination of carvacrol and amitraz on Demodex canis mites and investigate mites' survival lifetime. Methods: Mite samples were collected from a dog affected by demodicosis and had been diagnosed with deep skin scrapings. The motility of mites was examined by using optical microscopy. Samples were tested with 5% of carvacrol, the combination of Carvacrol 5% + Amitraz 0.05%, Amitraz 0.05% as a positive control, and mineral oil as a negative control. The interval time between the adding the test solutions and the moment the last mite ceased was defined as the survival time in the samples and the killing times of mites in each group were compared with each other. Results: It was determined that after administration of a 5% concentration of carvacrol, lethal effects were faster than the combination solution of Carvacrol 5% + Amitraz 0.05%, and the survival times in the control groups were longer than in both treatment groups. Conclusion: Carvacrol, as one of the most important bioactive terpenes, had the most effective in vitro miticidal activity, and it seems that carvacrol alone or in combination with amitraz can be suggested as a possible therapy in the treatment of canine demodicosis.

Determination of a Discriminant Dose to Identify Resistance to Amitraz in Rhipicephalus sanguineus s.l. (Acari: Ixodidae) from Mexico.

The brown dog tick, Rhipicephalus sanguineus s.l., is considered the most widely distributed three-host tick in the world and has medical and veterinary importance; the control of infestation is carried out with acaricides, towards which it can develop resistance. This study aimed to determine the discriminant dose (d.d) of amitraz to identify resistance in R. sanguineus s.l. larvae natives from Mexico and to evaluate its application in field-collected ticks. Engorged ticks were collected from naturally infested dogs residing in rural communities and were incubated for 25 days, and their progeny was used in a larval immersion test (LIT) to be exposed to the d.d. determined in Rhipicephalus microplus, and those that were susceptible were analyzed using the LIT in six concentrations. Mortality was analyzed through probit methodology to calculate the lethal concentration (LC) 50 and 99. The d.d. was determined as a consensus value by multiplying the LC99 × 2, and then, we proceeded to evaluate it in in-field samples by using the LIT technique. The d.d. calculated was 4 ppm. The in-field evaluation found 64% of the resistant samples to amitraz with mortality percentages between 98.3% and 0.35%. This dose can be used to rapidly and inexpensively identify resistant populations in samples collected in the field.

Acaricide Resistance Development in Rhipicephalus (Boophilus) microplus (Acari: Ixodidae) Populations against Amitraz and Deltamethrin on Communal Farms of the King Sabata Dalindyebo Municipality, South Africa.

Chemical acaricides are widely used to control ticks and tick-borne pathogens in cattle. However, prolonged and indiscriminate use of these chemicals inevitably leads to the selection of resistant ticks. In-vitro bioassays (adult and larval immersion tests) were conducted to assess amitraz and deltamethrin resistance in Rhipicephalus (Boophilus) microplus populations from communal farms of the King Sabata Dalindyebo municipality of South Africa. Data generated on percentage inhibition of oviposition (%IO) revealed that all the tick populations assessed showed resistance (%IO ≤ 95%) to at least one of the acaricides. All six tick populations assessed for efficacy (%IO ≥ 95%) at the DD) with deltamethrin were resistant (%IO ≤ 95%) and only one of the six tick populations assessed for efficacy with amitraz was susceptible. Based on the resistance ratios (RR), the adult immersion test detected amitraz and deltamethrin resistance in three (RR ranging from 2.30 to 3.21) and five (RR ranging from 4.10 to 14.59) of the six tick populations, respectively. With the larval immersion test, deltamethrin and amitraz resistance (larval mortality < 90% at the DD) was detected in all four and three of four R. (B.) microplus populations assessed, respectively. These data are critical for the design of an effective and sustainable tick control strategy on the communal farms.

Control of ticks and tick-borne diseases in Africa through improved diagnosis and utilisation of data on acaricide resistance.

A meeting, sponsored by the Bill and Melinda Gates Foundation (BMGF) and organised by Clinglobal, was held at The International Livestock Research Institute (ILRI) in Nairobi, Kenya, from 19th - to 21st October 2022. The meeting assembled a unique group of experts on tick control in Africa. Academia, international agencies (FAO and ILRI), the private Animal Health sector and government veterinary services were represented. The significant outcomes included: (i) a shared commitment to standardisation and improvement of acaricide resistance bioassay protocols, particularly the widely used larval packet test (LPT); (ii) development of novel molecular assays for detecting acaricide resistance; (3) creation of platforms for disseminating acaricide resistance data to farmers, veterinary service providers and veterinary authorities to enable more rational evidence-based control of livestock ticks. Implementation of enhanced control will be facilitated by several recently established networks focused on control of parasites in Africa and globally, whose activities were presented at the meeting. These include a newly launched community of practice on management of livestock ticks, coordinated by FAO, an African module of the World Association for the Advancement of Veterinary Parasitology (WAAVP-AN) and the MAHABA (Managing Animal Health and Acaricides for a Better Africa) initiative of Elanco Animal Health.

Evolution of acaricide resistance of Rhipicephalus decoloratus on commercial farms in South Africa.

The development of tick resistance to chemical control plays a major role in the increasing global economic impact of ticks on cattle farming. Reports on acaricide resistance of Rhipicephalus decoloratus, endemic to Africa and South Africa, are relatively few compared to the closely related and globally distributed Rhipicephalus microplus. In South Africa, ectoparasite control became the sole responsibility of each commercial producer when compulsory dipping was phased out from 1984. Different acaricidal management strategies resulted in the simultaneous development of resistance to various acaricide groups. The establishment of a Pesticide Resistance Testing Facility provided the opportunity to test Rhipicephalus (Boophilus) populations, submitted from all over South Africa, for resistance where failure of chemical control was experienced. The number of populations resistant to cypermethrin (CM) was significantly higher than those tested as resistant to amitraz (AM), or chlorfenvinphos (CFVP). No significant difference was found between the number of populations resistant to AM and CFVP. The evolution of R. decoloratus resistance at the end of a 12 year period indicated a stable but high prevalence of 90% overall resistance to CM. The same trend was seen for AM-resistant R. decoloratus populations but at a lower level of just over 40%. In contrast, CFVP resistant R. decoloratus populations showed a decreasing trend with near-total reversion to susceptibility. Multi-resistance was present in more than 50% of populations tested with the highest incidence in the Eastern Cape, KwaZulu-Natal, and Western Cape provinces.

Acaricide resistance of Rhipicephalus (Boophilus) decoloratus (Acari: Ixodidae) on commercial farms in South Africa: filling a gap in historical data.

In South Africa, acaricides are widely used for tick control but very few reports are available on resistance development of Rhipicephalus (Boophilus) decoloratus Koch to chemical control on commercial farming systems in Africa, south of the Sahara. Resistance to different acaricide classes reported over the years was mostly from localised communal farming systems. This report addresses the lack of available information on resistance development by reviewing results found during a National Tick Resistance Survey carried out from 1998 to 2001, laying the foundation for more recent research on resistance development, and the evolution of resistance over the years. One hundred and eighty R. decoloratus populations were randomly collected from commercial farming systems, covering most of the provinces of South Africa. Larval immersion tests were used to determine phenotypic resistance for each tick population and 6.6% of the populations tested were found to be resistant to amitraz, 35.5% to cypermethrin, and 36.1% to chlorfenvinphos. Multi-resistance to all three acaricides was found in 1.2% of populations and a further 25.8% of the populations were resistant to two acaricides. The detection of resistance of Rhipicephalus (Boophilus) species to currently used or new acaricides is an essential tool in resistance management. The acaricides tested for the resistance of R. decoloratus during the survey are currently still in use in South Africa and these historical results, never published before, can be valuable and can act as reference data to determine the evolution of resistance development to acaricides in more recent studies.

Assessment of cypermethrin and amitraz resistance and molecular profiling of voltage-gated sodium channel and octopamine tyramine genes of Rhipicephalus microplus.

Control of ticks and tick-borne pathogens is a priority for human and animal health. Livestock-holders extensively rely on acaricide applications for tick control. Different groups of acaricides including cypermethrin and amitraz have been consistently used in Pakistan. There has been a gap in understanding the susceptibility or resistance of Rhipicephalus microplus, the most prevalent tick in Pakistan, to acaricides. The present study aimed to molecularly characterize cypermethrin and amitraz targeted genes such as voltage-gated sodium channel (VGSC) and octopamine tyramine (OCT/Tyr) of R. microplus ticks in Khyber Pakhtunkhwa (KP), Pakistan to monitor the acaricides resistance. Tick specimens were collected from cattle and buffaloes in northern (Chitral, Shangla, Swat, Dir, and Buner), central (Peshawar, Mardan, Charsadda, Swabi, and Nowshera), and southern districts (Kohat, Karak, Lakki Marwat, Tank, and Dera Ismail Khan) of KP, Pakistan. Different concentrations of commercially available cypermethrin (10%) and amitraz (12.5%) were prepared for in vitro larval immersion tests (LIT). In LIT, the average mortality rate of immersed larvae was recorded that was increased gradually with an increase in the concentration of specific acaricide. The larvae's highest mortality rates (94.5% and 79.5%) were observed at 100-ppm of cypermethrin and amitraz, respectively. A subset of 82 R. microplus ticks was subjected to extract genomic DNA, followed by PCR to amplify partial fragments of VGSC (domain-II) and OCT/Tyr genes. The BLAST results of the consensus sequence of VGSC gene (domain-II) showed 100% identity with the acaricides susceptible tick sequence from the United States (reference sequence). Obtained identical sequences of OCT/Tyr genes showed maximum identity (94-100%) with the identical sequences reported from Australia (reference sequence), India, Brazil, Philippines, USA, South Africa, and China. Thirteen single nucleotide polymorphisms (10 synonymous and three non-synonymous) were observed at various positions of partial OCT/Tyr gene fragments. The SNP at position A-22-C (T-8-P) in OCT/Tyr gene has been linked to amitraz resistance in R. microplus ticks. Molecular analysis and LIT bioassay's findings indicate the availability of resistant R. microplus ticks in the KP region. To our understanding, this is the first preliminary study to monitor cypermethrin and amitraz resistance via molecular profiling of cypermethrin and amitraz targeted genes (VGSC and OCT/Tyr) in combination with in vitro bioassays (LIT) in R. microplus ticks from Pakistan.

Acaricidal efficacy evaluation of amitraz and diazinon against Amblyomma variegatum tick species in Waghimra zone, northern Ethiopia.

Ticks pose a substantial economic burden associated with production loss and treatment costs globally. Ethiopia has tremendous livestock resources, but its productivity is hindered by various animal health challenges in which ticks are the top priority, with a limited response to acaricidal treatments. Hence, we designed an acaricidal efficacy trial for the top commercially available Chemicals (amitraz and diazinon) to examine their efficacy against the widely distributed tick species (Amblyoma variegatum). Ticks were collected from animals admitted to veterinary clinics with no history of treatment with acaricides. Adult Immersion Technique (AIT) was employed to detect acaricidal resistance, and the mean percent control and antiparasitic efficacy were used to estimate tick susceptibility with a completely randomized laboratory-based trial (CRT). The mean weights of eggs laid by ticks subjected to amitraz and diazinon indicated that amitraz had a better egg-laying-inhibition effect than diazinon. The mean control percentages of amitraz and diazinon were 92.8 ± 5.6% and 69.7 ± 3.1%, respectively, with a highly significant difference (P-value = 0.00). The antiparasitic efficacy of the two drugs was 57.5 ± 0.96 and 37.5% ± 0.96 for amitraz and diazinon, respectively, which revealed that amitraz was statistically better than diazinon in killing adult ticks (P-value =0.026). In general, ticks treated with diazinon showed evidence of resistance development, and amitraz is relatively the most effective acaricide; we recommend its use in the study area and other locations with similar settings.

Transcriptome Profiling of Rhipicephalus annulatus Reveals Differential Gene Expression of Metabolic Detoxifying Enzymes in Response to Acaricide Treatment.

Ticks are hematophagous ectoparasites of economic consequence by virtue of being carriers of infectious diseases that affect livestock and other sectors of the agricultural industry. A widely prevalent tick species, Rhipicephalus (Boophilus) annulatus, has been recognized as a prime vector of tick-borne diseases in South Indian regions. Over time, the use of chemical acaricides for tick control has promoted the evolution of resistance to these widely used compounds through metabolic detoxification. Identifying the genes related to this detoxification is extremely important, as it could help detect valid insecticide targets and develop novel strategies for effective insect control. We performed an RNA-sequencing analysis of acaricide-treated and untreated R. (B.) annulatus and mapped the detoxification genes expressed due to acaricide exposure. Our results provided high-quality RNA-sequenced data of untreated and amitraz-treated R. (B.) annulatus, and then the data were assembled into contigs and clustered into 50,591 and 71,711 uni-gene sequences, respectively. The expression levels of the detoxification genes across different developmental stages of R. (B.) annulatu identified 16,635 transcripts as upregulated and 15,539 transcripts as downregulated. The annotations of the differentially expressed genes (DEGs) revealed the significant expression of 70 detoxification genes in response to the amitraz treatment. The qRT-PCR revealed significant differences in the gene expression levels across different life stages of R. (B.) annulatus.

Amitraz and Its Metabolites: Oxidative Stress-Mediated Cytotoxicity in HepG2 Cells and Study of Their Stability and Characterization in Honey.

The population decrease of bees that has been observed in recent years due to the Varroa destructor parasite may endanger the production of bee-products whose demand is on the rise. To minimize the negative effects caused by this parasite, the pesticide amitraz is commonly used by beekeepers. Based on these, the objectives of this work are to determine the toxic effects caused by amitraz and its metabolites in HepG2 cells, as well as its determination in honey samples and the study of its stability with different heat treatments commonly used in the honey industry and its relationship with the amount of 5-hydroxymethylfurfural (HMF) produced. Amitraz significantly decreased cell viability by MTT assay and total protein content (PC) assay, being more cytotoxic than its metabolites. Amitraz and its metabolites caused oxidative stress by Lipid Peroxidation (LPO) production and Reactive Oxygen Species (ROS) generation. Residues of amitraz and/or its metabolites were found in analyzed honey samples, with 2,4-Dimethylaniline (2,4-DMA) being the main metabolite confirmed by high-performance liquid chromatography-high resolution mass spectrometry (HPLC-QTOF HRMS). Amitraz and its metabolites resulted as unstable even at moderate heat treatments. Additionally, a positive correlation in terms of HMF concentration in samples and the severity of heat treatment was also observed. However, quantified amitraz and HMF were within the levels set in the regulation.

Target identification and acaricidal activity difference of amitraz and its metabolite DPMF in Tetranychus cinnabarinus (Boisduval).

BACKGROUND: Amitraz is a broad-spectrum formamidine acaricide proven to be effective against mites in all development stages. Under acidic conditions, amitraz is hydrolyzed to N2 -(2,4-dimethylphenyl)-N1 -methyformamidine (DPMF), an active metabolite for mite control. Octopamine and tyramine receptors are well known targets of amitraz. Until now, no research has been conducted about the amitraz target in Tetranychus cinnabarinus. This study aimed to identify the target genes of amitraz in T. cinnabarinus and reveal the mechanisms behind the differential acaricidal activities of amitraz and DPMF. RESULTS: Analysis of the toxicity, stress expression, target sensitivity and binding site of amitraz against T. cinnabarinus showed that TcOctß2R was the main target gene of amitraz. DPMF had more potent acaricidal activity against T. cinnabarinus and was more effective at activating TcOctß2R than amitraz. Furthermore, the three synergists had no significant effect on amitraz and DPMF, indicating that the detoxification metabolism was not related to the difference in acaricidal activity. CONCLUSION: In this study, TcOctß2R was identified as the main target gene of amitraz against T. cinnabarinus. The divergence of target binding was responsible for the difference in acaricidal activity between amitraz and DPMF. The results also revealed the physiological and pharmacological functions of octopamine receptors (OARs) in T. cinnabarinus and could provide a basis for the design of new acaricides, with OARs as a special target. © 2023 Society of Chemical Industry.

Prevalence of cattle ixodid ticks and acaricides efficacy evaluation in Amibara district of Afar Region, Ethiopia.

A cross sectional and randomized controlled trial study was conducted starting from July 2021 to July 2022 to determine the prevalence of ixodid ticks, and evaluate the efficacy of commonly used acaricides (amitraz 12.5% and diazinon 60%) in vitro and in vivo on cattle ticks in Amibara district. A total of 372 cattle were visited randomly from six kebeles of the district which were selected based on their livestock population. A total of 4330 adult ticks were collected and identified to the species level with stereomicroscope. For the invitro acaricide efficacy test, 90 engorged female ticks were collected from infested herds and grouped into three, each group having ten engorged ticks; two groups for the two test acaricides and the third groups were control to evaluate these acaricides in the laboratory. All the groups were replicated three times and incubated for seven days with similar temperature and humidity. Thirty-six infested cattle were selected and again grouped into three randomly, 12 each for the invivo test. The first groups of animals were sprayed with amitraz, the second with diazinon and the third groups were left untreated. Ticks were counted and recorded before, day 3, day 7, day 14 and day 21 of spraying. A total of 360 (96.7%) of the visited cattle were infested with ticks. Amblyomma. Rhipicephallus, Hyalomma and Bophillus were the genera of ticks identified with a prevalence of 44.9%, 41.4%, 7.5% and 6.2% respectively. A total of 15 species of ixodid ticks were identified from which Rhipicephallus pulchellus (39.86%) was dominant followed by Amblyomma lepidum (26.65%) and Amblyomma gemma (14.36%). Diazinon 60% EC and amitraz 12.5% were significantly inhibited the egg laying ability of engorged ticks' in vitro with percent inhibition of 100% and 99.1% respectively. On live animal, both acaricides showed statistically significant difference from the control group (p- value <0.05) with maximum 97.37% (amitraz) and 93.33% (diazinon) at day 7 and minimum 73.3% (amitraz) and 73.7% (diazinon) efficacy. However, there was no statistically significant difference on egg laying inhibition and tick count reduction between the two acaricides (p-value >0.05). Therefore, awareness should be created about the high prevalence as well as the application, dosage and dilution of the acaricides to pastoralists, community animal health workers and para veterinarians in the study area. Cattle owners, veterinarians and concerned government officials should address infested herds with manufacturer recommended dosage of these acaricides to control the tick infestation.

A Preliminary Study on "Personalised Treatment" against Varroa destructor Infestations in Honey Bee (Apis mellifera) Colonies.

The requirement for the protection of bee colonies against Varroa destructor invasions has been noted by many breeders and is included as an aspect of the development of beekeeping. This research aimed to check the effect of the development of a colony exposed to laying eggs (brood surface) by queen bees with similar chemical potential (sisters) on the effect of a preparation combating V. destructor depending on the number of mites found in a given colony. We chose this as a standard model of conduct that treats each bee colony as one organism subjected to individual parasite control. For this purpose, we created a bee colony with a mother-of-one breeding line and fertilised drones from one colony. Infection with V. destructor occurred naturally and uncontrollably. Without interfering with the colony's development, the frame insulator helped each colony's brood (mothers' reproductive potential) and the initial and final individuals from the mites themselves. The study was carried out in four species (two control species and two species with up to 20 and over 21 mites, respectively). Treatments with amitraz to combat damage were divided into four treatment subgroups: two treatments every four days or four treatments every two days. We observed the number of individuals that were protected in all subgroups in the average brood area. The reproductive potential of the sisters' mothers did not change after the treatments with amitraz, which indicated that amitraz did not affect the delegation of egg laying. The invasion rate was also tracked relative to the control group, which allowed us to conclude that a two-time treatment with amitraz reduced the frequency of mites and a four-time treatment checked the effectiveness. Tailoring the control of V. destructor in bee colonies may be an effective measure in the fight against this parasite.