Ticks are important ectoparasites in equids, causing economic losses in animal husbandry in Iran and worldwide. This study was aimed to determine frequency and species diversity of hard ticks in equids in Ardabil province, during the four seasons in 2021. A total of 240 equids (187 horses, 53 donkeys) were randomly selected and examined. Ixodid ticks were collected from body surface of examined animals and identified. Of all examined equids, 32.5% horses, and 4.58% donkeys were infested with a total number of 412 ixodid ticks. Tick indices (tick number per animal) were 4.62. There was significant difference between prevalence and different age groups of infested animals. The highest prevalence of hard ticks was found in ≥3 years-old equids in the region. Of all examined ticks (412), three genera including Hyalomma spp., Rhipicephalus spp. .and Dermacentor spp. with four species. i.e. Hyalomma anatolicum (73.05%), H. marginatum (17.23%), Rhipicephalus bursa (7.76%) and Dermacentor marginatus (1.94%) were identified. Ixodid ticks\' infestations occurred throughout the year with the highest prevalence in spring. The findings indicated that the equids had a high species diversity and ixodid tick infestation, necessitating additional research to identify any possible tick vectors for the hemoparasite transmission in equids of this region.

Hard ticks pose a threat to animal and human health. Active life stages need to feed on a vertebrate host in order to complete their life cycle. To study processes such as tick-pathogen interactions or drug efficacy and pharmacokinetics, it is necessary to maintain tick colonies under defined laboratory conditions, typically using laboratory animals. The aim of this study was to test a membrane-based artificial feeding system (AFS) applicable for Amblyomma ticks using Amblyomma tonelliae as a biological model. Adult ticks from a laboratory colony were fed in a membrane-based AFS. For comparison, other A. tonelliae adults were fed on calf and rabbit. The proportions of attached (AFS: 76%; calf/rabbit: 100%) and engorged females (AFS: 47.4%; calf/rabbit: 100%) in the AFS were significantly lower compared to animal-based feeding (p = 0.0265). The engorgement weight of in vitro fed ticks (x¯ = 658 mg; SD ± 259.80) did not significantly differ from that of ticks fed on animals (p = 0.3272, respectively 0.0947). The proportion of females that oviposited was 100% for all three feeding methods. However, the incubation period of eggs (x¯ = 54 days; SD ± 7) was longer in the AFS compared to conventional animal-based feeding (p = 0.0014); x¯ = 45 days; SD ± 2 in the rabbit and (p = 0.0144). x¯ = 48 days; SD ± 2 in the calf). Egg cluster hatching (x¯ = 41%; SD ± 44.82) was lower in the AFS than in the other feeding methods (rabbit: x¯ = 74%; SD ± 20; p = 0.0529; calf: x¯ = 81%; SD ± 22; p = 0.0256). Although the attachment, development, and the hatching of AFS ticks were below those from animal-based feeding, the method may be useful in future experiments. Nevertheless, further experiments with a higher number of tick specimens (including immature life stages) and different attractant stimuli are required to confirm the preliminary results of this study and to evaluate the applicability of AFS for Amblyomma ticks as an alternative to animal-based feeding methods.