A One-Month Survey on Infestation of Ixodidae (Acari: Ixodida) Ticks Collected from Dogs in the Robat Karim Region, Tehran Province, Iran

1. Introduction

The increasing number of dogs in the country and the increase in the presence of stray dogs in cities, as well as the l incomplete implementation of health regulations, has heightened the risk of transmitting common diseases from dogs to humans. The most important and common diseases in dogs are those caused by ectoparasite arthropods. Arthropods represent over 80% of all organisms on the planet and can act both as external parasites and as vectors for parasitic, viral, and bacterial infections ( 1 ). Ticks are among the most important arthropods in the world, causing high damages in the veterinary field of many countries each year. Therefore, after mosquitoes, ticks are are considered the second most important group of arthropods in the veterinary medicine, particularly in tropical countries ( 2 , 3 ). Ixodidae ticks are the most important and common carriers of pathogens among ticks and are recognized as one of the most important external parasites affecting dogs in the world ( 4 ). Ixodidae ticks can cause a lot of damage to dogs, including blood loss, dermatitis, pain, and a variety of parasitic, bacterial, and viral infections such as tick-borne encephalitis virus, Ehrlichia canis, and Babesia canis ( 5 , 6 , 7 ). According to the surveys, there are about 700 million domestic dogs in the world, and 75% of this population are stray dogs ( 8 ). Today, due to the increasing presence of stray dogs in cities and their proximity to human communities, they are considered one of the biggest problems for public health ( 9 ).

Therefore, it is very important to determine the distribution and prevalence of ticks among all dogs, especially stray dogs ( 10 , 11 ). As the capital of Iran, Tehran province is considered the most important and most populated region of Iran. With the growing population in Tehran, tend to settle in the surrounding areas, such as the Robat Karim region, which is close to Tehran. As the capital expands, the population of Robat Karim is also increasing. Therefore, investigating the risk factors of disease in this region is more important than ever.

2. Case Presentation

2.1. Study Area

Robat Karim region is located in the southwestern region of Tehran province, at a longitude of 51:4, latitude of 35:28 and an altitude of 1100 meters above the sea level. The area of this city is about 275 square kilometers, bordered by Ray and Islamshahr from the south, Shahriar to the north, Baharestan to the east, and Zarandieh to the west. The annual rainfall of this area is about 200 ml. The maximum temperature reaches 44 degrees Celsius ,while the minimum temperature drops to -20 degrees Celsius. Overall,the average air temperature in Robat Karim is 16 degrees Celsius. Due to its proximity to the capital of Iran, the population of this region is growing, with about 291,515 residents living in this region (Figure 1).

Figure 1. Map of Iran showing the location of Tehran province and Rabat Karim region.

2.2. Data Collection

This study was conducted in 4 urban areas (Alard, Nasirshahr, Parand and Robat Karim) and 10 rural areas (Peyghambar, Anjemabad, Manjilabad, Hoseynabad-e Yangejeh, Shahrabad-e Ilat, Asgharabad, Laqeh Hesar Mehtar, Vahnabad and Hakimabad), from September 1st to September 30^th, 2023 (Table 1). A total of 83 dogs (17 herd dogs and 66 stray dogs) were examined using random-cluster sampling. From these dogs, 434 ticks were detected accross 5 parts of the dog's body. In this method, the dog's body was divided into 5 parts: I. head, ears and neck; II. Dorsal; III. Abdomen, groin, axillary and inguinal; IV. Legs and feet and V. tail and perianal ( 12 ).

All sampling was done between 8:00 am and 12:00 pm. The age of the studied dogs was determined by asking the owner and assessing to the dental formula. Ticks were collected from the dogs' body using forceps slowly at a 45- degree angle and placed inside the numbered tubes containing 70% ethanol. These samples were sent to the Entomology Laboratory of Bu - Ali University ,Faculty of Agriculture, for further examination and clarification. They were transferred to Sinai Hamadan. A total of 279 Ixodidae ticks were identified using a stereomicroscope with a magnification of 40 to 80 times and compared with valid keys ( 13 , 14 ).

2.3. Preparation of Slides and Clarification Of Ticks

The tick samples preserved in ethanol were washed to remove any adherent host tissue and then placed in glass vials. Blood contents of the ticks were drained from the abdominal area of the ticks using a syringe needle. The ticks were placed in 10% potassium hydroxide to dissolve unwanted chitin and debris, making them clear and clean for microscopic examination. Tick samples were washed several times with water and then dehydrated through successive dilutions of ethyl alcohol (70, 80, 90, 95 and 100). They were then clarified and cleared using xylene for 15 to 30 minutes.

Tick samples were mounted on glass slides using Canadian balsam glue, covered with a coverslip, to dry at laboratory temperature and finally examined under a light microscope ( 15 , 16 ) (Figure 2).

Figure 2. Tick samples isolated from dog, a. Oral appendages of Hyalomma marginatum b. Oral appendages of Rhipicephalus sanguineus c. Hyalomma marginatum d. Rhipicephalus sanguineus

2.4. Statistical Analysis

The Chi-square test (2χ) was used to statistically analyze the obtained data and determine the relationship between the prevalence of infection by different species of ticks isolated with age, gender and place of isolation. Also, at first, the data collected from the isolation site was entered into Microsoft Excel 2016 software.

Final statistical analysis was performed using SPSS 2021 software, with a significance level of p≤0.05.

3. Results

3.1. Species diversity of Ixodidae ticks

Out of 83 examined dogs, 72 dogs were found to be infected with Ixodidae ticks. A total of 434 Ixodidae ticks, 2 genera of Rhipicephalus and Hyalomma and 4 species of Rhipicephalus sanguineus, Rhipicephalus bursa, Rhipicephalus turanicus, Hyalomma marginatum were detected. Among these, 279 (64.28%) of the identified Ixodidae ticks belonged to Rhipicephalus sanguineus, which can be said to be the most common tick species in Robat Karim region in Tehran province. This was followed by 75 (17.28%) Rhipicephalus bursa, 49 (11.29%) Rhipicephalus turanicus and 31 (7.14%) Hyalomma marginatum of Ixodidae respectively (Table 2).

3.2. Geographical Distributions

The geographical distribution of ticks on dogs showed that Rhipicephalus sanguineus is the most common species, while Hyalomma marginatum is the least common species among all investigated areas in Rabat Karim region of Tehran province. In the region of Rabat Karim, the cities of Nasirshahr, Parand, Robat Karim and the villages of Anjemabad, Hoseynabad-e Yangejeh, Shahrabad-e Ilat, Vahnabad, Aliabad, Kazemabad, recorded the highest percentage of infection , with all the dogs examined in these areas were infected with Ixodidae ticks. Laqeh village was the only part that did not record any infestation with Ixodidae ticks. Out of a total of 434 Ixodidae ticks isolated from dogs in Robat Karim region, 153 ticks were in urban areas and 281 ticks were in rural areas, which indicates that tick infestation in dogs in rural areas is higher than in urban areas. The highest number of ticks in the urban area of Parand city with 50 (32.67%), while Aliabad village had the highest count in rural areas, with 56 (19.92%) number of Ixodidae ticks recorded (Table 3).

4. Discussion

In this study, which was the first specialized investigation on Ixodidae ticks in the Robat Karim region of Tehran province, 2 different genera of Ixodidae ticks, Rhipicephalus and Hyalomma, were identified, along with 4 species Rhipicephalus sanguineus, Rhipicephalus bursa, Rhipicephalus turanicus and Hyalomma marginatum. The findings indicated that the predominant tick species in the Robat Karim region is Rhipicephalus sanguineus, accounting for 64.28% of the total ticks identified. This prevalence is approximately equal to the research conducted in Argentina with a prevalence rate of 73% and Thailand with a prevalence rate of 74.20% ( 17 , 18 ).

Additionally, studies conducted in Iran, in Ilam province (27.50%) and the Gilanegharb region in Kermanshah province (35.36%), also identified Rhipicephalus sanguineus as the dominant tick species ( 19 , 20 ). The number of ticks found on male dogs compared to female dogs shows a significant difference, with 284 (65.44%) ticks found on male dogs and 150 (34.56%) on female dogs. However, overall infestation rates for external parasites are 85.71% in male dogs and 88.88% in female dogs. Regarding age-related variables, out of the total of 434 Ixodidae ticks, 192 (44.23%) were related to dogs aged 1-3 years, while the lowest number, 53 ticks (12.21%) belonged to the dogs under 1 year. The intensity of infestation in stray dogs compared to owned dogs does not show a significant difference: 82.35% of owned dogs and 87.87% of stray dogs were infested with Ixodidae ticks. On average, 8.5 ticks per dog were identified on owned dogs, while stray dogs had 5.43 ticks per dog, showing a lower number compared to a study conducted by Yi Yan and colleagues in 2023, where Malaysia reported an infestation rate of 8.13 ticks per dog, and the Philippines reported 25.75 ticks per dog for stray dogs ( 21 , 22 ) (Table 4). Some studies, reported by different researchers, have also isolated dog ixodid ticks from the skin of other animals, such as cats and ruminants ( 23 - 26 ).

The analysis showed that specific body parts significantly affect the presence of ticks in dogs. Of the total ticks collected, 211 (48.61%) ticks were found in the Abdomen, axillary, groin, and inguinal region, 70 (16.12%) in the Legs and feet region, 67 (15.43%) in the dorsal region, 51 (11.75%) There were 35(8.06%) in Tail and perianal area and 35(8.06%) in Head, ears and neck area, which shows that Abdomen, axillary, Groin, inguinal area is the most heavily infected area in the body of dogs in Robat Karim area with Ixodidae ticks. (Table 5).

Examining the infected areas of the dogs' body with different types of ticks found shows that the highest number of Rhipicephalus sanguineus and Rhipicephalus bursa were in Abdomen, axillary, groin, inguinal region, while the highest number of Rhipicephalus turanicus and Hyalomma marginatum were in dorsal region (Figure 3).

Figure 3. Distribution of attachment sites of adult ticks on dogs. A) Rhipicephalus sanguineus. B) Rhipicephalus bursa. C) Rhipicephalus turanicus. D) Hyalomma marginatum.

This study provides valuable insights into Ixodidae tick infestation and associated risk factors in herding and stray dogs. Adapting preventive strategies and interventions based on sensitive and vulnerable body parts can more effectively protect dogs against ticks and reduce health risks. As in this study, variables such as gender and age have been examined in detail, which helps implememt controlling and preventive plans in the region. Our study showed that Rhipicephalus sanguineus, as a tick of tropical lineage, is the dominant tick in Robat Karim area of Tehran province.

However, this study did not assess seasonal prevalence due to limitations, nor did it evaluate the pathogens transmitted by Ixodidae ticks. Such studies conducted on larger scales and accross larger regions of Iran can help to identify the tick fauna specific to each region and control the biological problems in that region. Studies similar to our work should be done in other regions of Iran to determine the national level of tick infestation in dogs and the results of these studies can be used in strategic tick control programs.

Acknowledgment

The authors hereby extend their gratitude to all the technicians and experts of the parasitology laboratory who helped us in collecting and preparing tick samples.

Authors' Contribution

Study concept and design: G.A, G.Y.

Acquisition of data: G.A, D.E.

Analysis and interpretation of data: G.A, G.Y, D.E.

Drafting of the manuscript: G.A, G. Y.

Critical revision of the manuscript for important: G.Y.

Intellectual content: G.A, D.E.

Sampling: G.A, D.E, J, M.

Statistical analysis: G.A, D.E, J. M.

Ethics

All principles of medical ethics have been observed in this study.

Conflict of Interest

The authors declare no competing interests.

Funding

Not applicable.

Data Availability

The data that support the findings of this study are available on request from the corresponding author.

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