Molecular investigation on transstadial/transovarial transmission of Hepatozoon canis in Rhipicephalus sanguineus sensu lato ticks

Bartonella henselae is a slow-growing, Gram-negative bacterium that causes cat scratch disease in humans. A transstadial transmission of the bacteria from larvae to nymphs of Rhipicephalus sanguineus sensu lato (s.l.) ticks, suspected to be a potential vector of the bacteria, has been previously demonstrated. The present study aims to investigate transovarial transmission of B. henselae from R. sanguineus s.l. adults to their instars. Adult ticks (25 males and 25 females) were fed through an artificial feeding system on B. henselae-infected goat blood for 14 days, and 300 larvae derived from the experimentally B. henselae-infected females were fed on noninfected goat blood for 7 days. Nested PCR and culture were used to detect and isolate B. henselae in ticks and blood samples. Bartonella henselae DNA was detected in midguts, salivary glands, and carcasses of the semi-engorged adults and pooled tick feces (during feeding and post-feeding periods). After the oviposition period, B. henselae DNA was detected in salivary glands of females (33.3%), but not in pooled eggs or larvae derived from the infected females. However, B. henselae DNA was detected by nested PCR from the blood sample during larval feeding, while no viable B. henselae was isolated by culture. According to our findings, following infected blood meal, B. henselae could remain in the tick midguts, move to other tissues including salivary glands, and then be shed through tick feces with limited persistency. The presence of bacterial DNA in the blood during larval feeding shows the possibility of transovarial transmission of B. henselae in R. sanguineus s.l. ticks.

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First molecular evidence of Hepatozoon canis infection in Rhipicephalus sanguineus (sensu lato) ticks in Iran

10.21203/rs.3.rs-81343/v1 ◽

2020 ◽

Author(s):

MohammadReza Zeinali ◽

Farnaz Malekifard ◽

Alaleh Rakhshanpour ◽

Mohammad Yakhchali

Keyword(s):

Rhipicephalus Sanguineus ◽

Diagnostic Methods ◽

Ixodid Ticks ◽

Hepatozoon Canis ◽

Molecular Evidence ◽

Microscopical Examination ◽

Serum Samples ◽

Blood Samples ◽

Significant Difference ◽

Rhipicephalus Sanguineus Sensu Lato

Abstract Background Hepatozoon canis is a protozoan that is transmitted by the ixodid ticks. Ingesting the tick or a section of the tick organ which is infected by the mature oocysts containing infectious sporozoite is the main source of infection in dogs. Canine hepatozoonosis infects dogs in Iran, but the vector for Hepatozoon protozoa from Iran has never been demonstrated. The present study aims to detect H. canis in dogs and vector ticks in Iran. Methods During the period of 2018–2019, Blood samples and ixodid ticks were collected and examined using microscopical, molecular, and serological methods. A total of 246 blood samples were collected from the cephalic vein of pet, stray, and shelter dogs (103 stray, 99 shelter, and 44 pets) of both genders and varying ages in Northwest of Iran. Results Microscopically, infected neutrophils with Hepatozoon spp. were detected in 5 of 246 (2.03%) thin stained blood smears with low parasitemia. Indirect immunofluorescent antibody test (IFAT) was used to test the serum samples and antibodies against H. canis were detected in 31 (12.6%) of the serum samples. Molecularly, 23 out of 246 (9.34%) blood samples were found to be infected with H. canis. A comparison of the results of 3 diagnostic methods demonstrated a good agreement between IFAT and PCR and a poor agreement between microscopical examination with IFAT and PCR. There was no significant difference in different age groups and sex of sampled dogs. However, stray dogs had significant infection rate of than pets and shelter ones. In body inspection, 141 adult ticks (31 partially engorged females, 26 fully engorged female and 84 fed males) were collected from examined dogs and all ticks were belonging to species of Rhipicephalus sanguineus (sensu lato). Positive reaction to H. canis was observed in the genomic DNA of the 7 ticks (4.96%). A BLAST analysis of obtaining sequences isolated from both dogs and ticks indicated a 99–100% similarity with H. canis 18S rRNA gene sequences in GenBank. This is the first study in Iran to detect H. canis in R. sanguineus tick.

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First detection and genetic identification of Hepatozoon canis in Rhipicephalus sanguineus sensu lato ticks collected from dogs of Taiwan

Ticks and Tick-borne Diseases ◽

10.1016/j.ttbdis.2019.04.020 ◽

2019 ◽

Vol 10 (4) ◽

pp. 929-934 ◽

Cited By ~ 1

Author(s):

Li-Lian Chao ◽

Hsin-Ting Liao ◽

Chien-Ming Shih

Keyword(s):

Rhipicephalus Sanguineus ◽

Genetic Identification ◽

Hepatozoon Canis ◽

Rhipicephalus Sanguineus Sensu Lato

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Molecular investigation of bacterial and protozoal pathogens in ticks collected from different hosts in Turkey

Parasites & Vectors ◽

10.1186/s13071-021-04779-2 ◽

2021 ◽

Vol 14 (1) ◽

Author(s):

Ahmet Efe Köseoğlu ◽

Hüseyin Can ◽

Mervenur Güvendi ◽

Sedef Erkunt Alak ◽

Çağrı Kandemir ◽

...

Keyword(s):

Dna Isolation ◽

Rhipicephalus Sanguineus ◽

Hepatozoon Canis ◽

Molecular Method ◽

Anaplasma Ovis ◽

Babesia Caballi ◽

Molecular Investigation ◽

Tick Borne Disease ◽

Pcr Method ◽

First Time

Abstract Background The emergence of tick-borne disease is increasing because of the effects of the temperature rise driven by global warming. In Turkey, 19 pathogens transmitted by ticks to humans and animals have been reported. Based on this, this study aimed to investigate tick-borne pathogens including Hepatozoon spp., Theileria spp., Babesia spp., Anaplasma spp., Borrelia spp., and Bartonella spp. in tick samples (n = 110) collected from different hosts (dogs, cats, cattle, goats, sheep, and turtles) by molecular methods. Methods To meet this objective, ticks were identified morphologically at the genus level by microscopy; after DNA isolation, each tick sample was identified at the species level using the molecular method. Involved pathogens were then investigated by PCR method. Results Seven different tick species were identified including Rhipicephalus sanguineus, R. turanicus, R. bursa, Hyalomma marginatum, H. anatolicum, H. aegyptium, and Haemaphysalis erinacei. Among the analyzed ticks, Hepatozoon spp., Theileria spp., Babesia spp., and Anaplasma spp. were detected at rates of 6.36%, 16.3%, 1.81%, and 6.36%, respectively while Borrelia spp. and Bartonella spp. were not detected. Hepatozoon spp. was detected in R. sanguineus ticks while Theileria spp., Babesia spp., and Anaplasma spp. were detected in R. turanicus and H. marginatum. According to the results of sequence analyses applied for pathogen positive samples, Hepatozoon canis, Theileria ovis, Babesia caballi, and Anaplasma ovis were identified. Conclusion Theileria ovis and Anaplasma ovis were detected for the first time to our knowledge in H. marginatum and R. turanicus collected from Turkey, respectively. Also, B. caballi was detected for the first time to our knowledge in ticks in Turkey.

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