Nimo‑Paintsil et al. Parasites & Vectors (2022) 15:86 https://doi.org/10.1186/s13071‑022‑05208‑8 Parasites & Vectors RESEARCH Open Access Ticks and prevalence of tick‑borne pathogens from domestic animals in Ghana Shirley C. Nimo‑Paintsil1*†, Mba Mosore1,2†, Seth Offei Addo1,2, Taylor Lura3, Janice Tagoe1,2, Danielle Ladzekpo2, Charlotte Addae2, Ronald E. Bentil1,2, Eric Behene1,2, Courage Dafeamekpor4, Victor Asoala5, Anne Fox1, Chaselynn M. Watters1, Jeffrey W. Koehler6, Randy J. Schoepp6, Hanayo Arimoto7, Samuel Dadzie2*, Andrew Letizia8 and Joseph W. Diclaro II3 Abstract Background: Ticks are important vectors of various pathogenic protozoa, bacteria and viruses that cause serious and life‑threatening illnesses in humans and animals worldwide. Estimating tick‑borne pathogen prevalence in tick popu‑ lations is necessary to delineate how geographical differences, environmental variability and host factors influence pathogen prevalence and transmission. This study identified ticks and tick‑borne pathogens in samples collected from June 2016 to December 2017 at seven sites within the Coastal, Sudan and Guinea savanna ecological zones of Ghana. Methods: A total of 2016 ticks were collected from domestic animals including cattle, goats and dogs. Ticks were morphologically identified and analysed for pathogens such as Crimean‑Congo haemorrhagic fever virus (CCHFV), Alkhurma haemorrhagic fever virus (AHFV), Rickettsia spp. and Coxiella burnetii using polymerase chain reaction assays (PCR) and sequence analysis. Results: Seven species were identified, with Amblyomma variegatum (60%) most frequently found, followed by Rhipicephalus sanguineus sensu lato (21%), Rhipicephalus spp. (9%), Hyalomma truncatum (6%), Hyalomma rufipes (3%), Rhipicephalus evertsi (1%) and Rhipicephalus (Boophilus) sp. (0.1%). Out of 912 pools of ticks tested, Rickettsia spp. and Coxiella burnetii DNA was found in 45.6% and 16.7% of pools, respectively, whereas no CCHFV or AHFV RNA were detected. Co‑infection of bacterial DNA was identified in 9.6% of tick pools, with no statistical difference among the ecozones studied. Conclusions: Based on these data, humans and animals in these ecological zones are likely at the highest risk of exposure to rickettsiosis, since ticks infected with Rickettsia spp. displayed the highest rates of infection and co‑infec‑ tion with C. burnetii, compared to other tick‑borne pathogens in Ghana. Keywords: Tick‑borne pathogens, Livestock, Ghana, West Africa *Correspondence: snimo‑paintsil@noguchi.ug.edu.gh; shirleycameron93@yahoo.com; sdadzie@noguchi.ug.edu.gh †Shirley C. Nimo‑Paintsil and Mba Mosore contributed equally to this work 1 United States Naval Medical Research Unit No. 3, Ghana Detachment, Accra, Ghana 2 Noguchi Memorial Institute for Medical Research, College of Health Sciences, University of Ghana, Legon, Accra, Ghana Full list of author information is available at the end of the article © The Author(s) 2022. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecom‑ mons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data. Nimo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 2 of 11 Background of animal to human transmission and infection of Q Ticks are important vectors of various pathogenic pro- fever [15, 16]. The bacterium is found worldwide with tozoa, bacteria and viruses that cause morbidity and the exception of Antarctica and New Zealand and has mortality in humans and animals worldwide [1]. Domes- been documented in more than 40 species of ticks [17]. tic animals are parasitized by many tick species thereby However, limited information is available for C. burnetii causing considerable economic loss [2–4]. Human trans- prevalence in sub-Saharan Africa. In Kenya, one study mission of tick-borne diseases can occur through the bite demonstrated C. burnetii antibody prevalence of 10–20% of an infected tick, exposure to an infected animal, or in humans, and another showed C. burnetii antibody consuming animal products [5]. Evidence suggests that prevalence of 7–57% and 33–34% in domestic cattle and zoonotic tick-borne diseases are increasing in geographi- goats, respectively [18, 19]. In Ghana, C. burnetii has cal range, and infection rates are likely to become a major been detected in children and livestock in three different public health threat in the future [6]. regions [20–22]. Worldwide, ticks serve as important vectors of Most rickettsial pathogens are transmitted by ectopar- Crimean-Congo haemorrhagic fever virus (CCHFV) [5], asites during feeding or by scratching crushed infectious with species of the genus Hyalomma considered the prin- arthropods or infectious faeces into the skin. Several cipal vectors [7]. Wild and domestic animals such as cat- pathogenic tick-borne Rickettsia species have been found tle, sheep and goats play the role of amplifying hosts or in Africa including in Senegal, Burkina Faso, Cameroon, reservoirs in the spread of the virus [8]. Although human Mali and Ivory Coast, with human seroprevalence rates infections normally occur through tick bites, other pos- ranging from 17 to 36% [23, 24]. In travellers, including sible routes include drinking unpasteurized milk from military personnel, the most commonly diagnosed rick- infected animals and being exposed to blood or tissues ettsial diseases are usually spotted fever (African tick- from infected individuals or animals infected with the bite fever [ATBF]) or typhus groups (murine typhus), virus [9]. CCHFV is endemic to Africa, the Balkans, the but travellers may acquire a wide range of rickettsioses, Middle East and Asian countries, with a high case fatal- including emerging and newly recognized species [25]. ity rate [10]. In the Ashanti region of Ghana, which lies Military members train and deploy in numerous ter- within the deciduous forest, the virus has been detected rains hospitable to tick populations. Ticks harbour the in Amblyomma variegatum and Hyalomma excavatum aforementioned viral and bacterial pathogens that can ticks collected from cattle at the abattoir, with a seroprev- incapacitate or kill individual troops or cause an outbreak alence rate of 5.7% in animal handlers [11]. in a region and disrupt force health protection. Surveil- Alkhurma haemorrhagic fever virus (AHFV), a tick- lance of tick species in Ghana informs potential vector- borne flavivirus, was originally isolated in 1995 from a borne infectious threats for force health protection, patient in Saudi Arabia. Subsequent cases of AHF have improves planning for combatant commands, supports been documented in tourists in Egypt, indicating a wider in-country partners and promotes global security. geographical distribution of the virus [12]. Surveillance Vector-borne diseases remain a significant cause for this pathogen is supported by the wide distribu- of infection throughout the world, but information tion of AHFV tick hosts, namely, the soft tick Ornitho- regarding the risk of tick-borne infections is limited in doros savignyi and the hard tick Hyalomma dromedarii. Africa. Many tick-borne illnesses in humans are associ- A recent study on AHFV in ticks infesting migratory ated with domestic animals, particularly livestock [23, birds in transit from Africa to Europe and Asia, as well as 26, 27]. Ghana imports live animals, such as livestock other cases of seropositivity in Djibouti, could indicate a from neighbouring countries. This movement of ani- wider geographical distribution of the virus throughout mals may aid in the transmission of tick-borne disease eastern Africa and possibly the sub-Saharan region [13]. into Ghana. The spread of disease from livestock trade However, the persistence of the virus within tick popula- and migration patterns is compounded by the asympto- tions, the role of livestock and the disease transmission matic presentation of some tick-borne diseases in cattle, process are poorly understood, especially in Ghana and hindering the ability of inspectors to spot infected ani- West Africa. mals [16, 28, 29]. Additionally, livestock are commonly Coxiella burnetii, the causative agent of Q fever, is a allowed free movement to search for water and food. bacterial pathogen that causes abortion in livestock and These free-roaming animals have increased exposure is primarily transmitted to humans through infected ani- to various pathogens that in turn may be transmitted to mal birth products but is also transmitted by ticks [14]. livestock handlers, veterinarians, abattoir workers and Domestic ruminants represent the most frequent source the general population [30]. Ghana is a coastal country N imo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 3 of 11 bordered by three countries and offers different ecologi- Nucleic acid extraction and pathogen detection cal zones that may influence the distribution of arthro- Pooled ticks were homogenized using Mini-Bead- pod vectors and their diseases. Thus, this study sought beater-96 (Biospec, Bartlesville, OK, USA) with lysis to examine the prevalence of tick-borne pathogens buffer and beads of 0.1 mm and 2.0 mm diameter. Nucleic to better understand the public health risk in Ghana acid was extracted from each pool using the QIAamp and the West African subregion. The scientific benefit Viral RNA Mini Kit (Qiagen, Valencia, CA, USA) follow- includes the augmentation of knowledge regarding the ing the manufacturer’s instructions [32]. ecology of tick-borne pathogens circulating in the dif- The presence of viral RNA for CCHFV and AHFV ferent ecological zones. was detected using real-time reverse transcriptase poly- merase chain reaction (RT-PCR) as described previ- ously [12, 32, 33]. Briefly, 5 ml of extracted nucleic acid Methods was tested in duplicate by real-time RT-PCR using assays Study sites for CCHFV and AHFV using the SuperScript One-Step One civilian and six military sites within three ecological RT-PCR kit (Thermo Fisher Scientific). Synthetic RNAs zones of Ghana were selected for tick collection. Three (BioSyn, Inc., TX, USA) for the CCHFV assay amplicon of the military sites were within the Coastal savanna of and the AHFV assay amplicon were used as positive con- Accra; the other three were located in Tamale, all in the trols, and molecular biology-grade water was used as a Guinea savanna. The one civilian site was located in the negative template control. The positive and negative con- Sudan savanna (Fig. 1). Ticks were collected from cattle, trols were run on each real-time PCR plate. Fluorescence goats and dogs in surrounding communities near military readings were taken following each real-time PCR cycle, bases/camps. Livestock at the study sites were checked and a sample was considered positive if the quantification for tick infestation. cycle (Cq) value was less than 40 cycles. A sample was The Guinea and Sudan savanna sites were chosen indeterminate if there was an appropriate curve with a because of their proximity to Burkina Faso, Ivory Coast, Cq value of greater than 40 cycles, and the sample testing Togo and Mali. The increased populations of nomadic was repeated. The bacterial deoxyribonucleic acid (DNA) Fulani herdsmen whose occupation is commercial live- of Rickettsia spp. and C. burnetii was detected separately stock rearing results in more variety in livestock. How- using quantitative real-time PCR with Platinum TaqDNA ever, the influx of livestock from neighbouring countries Polymerase (Applied Biosystems, Thermo Fisher Sci- has the potential of introducing other tick species as well entific, Waltham, MA, USA) assay with sets of primers as tick-borne pathogens into the country. In the Coastal targeting the gene encoding the 17-kilodalton antigen savanna sites, most people keep domestic animals such (17-kDa) of Rickettsia DNA and the com1a gene of C. as cats, dogs and chickens alongside cattle for general burnetii, respectively [33, 34]. Aliquots of double distilled personal use as opposed to commercial purposes. Herds- water were included in all PCR runs to detect contami- men most often have dogs to accompany livestock during nation. The PCR assays were carried out on an ABI 7300 grazing, increasing potential exposure to tick infestation. Thermal Cycler (Applied Biosystems, Thermo Fisher Sci- entific, Waltham, MA, USA). Tick collection Tick collection was conducted between June 2016 and December 2017. Collection was performed in June and Statistical analysis December of 2016 and in March, July, August, Septem- Tick distribution was described using descriptive sta- ber and December of 2017. Verbal consent was sought tistics with frequency, percentages and bar graphs. from animal handlers prior to examining their livestock The infection rate was estimated using the frequen- for ticks. Using blunt forceps, ticks were collected (from tist approach [35]. For unequal pool size and under the the abdomen, neck, internal sides of rear legs, tail and assumption of a perfect test, maximum likelihood (ML) ear) and placed into labelled vials containing RNAlater™ was used to estimate the infection rate. A 95% Wald-type (Qiagen, Germany) ribonucleic acid (RNA) stabilizing confidence interval was reported for the infection rate. reagent. All ticks were morphologically identified with Statistical analysis was done using R version 3.3.0 soft- taxonomic keys [31]. Specimens were pooled by species, ware. Pearson Chi-square or Fisher’s exact test, where sex, study site and animal host. Pooled samples consisted necessary, was used to determine the association between of between one to five ticks. tick species and ecological zones. The association of Nimo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 4 of 11 Fig. 1 Map of Ghana displaying study sites, and geographical regions. Study sites—[Navrongo, Air Force Base (AF), 6th Battalion Infantry (6 BN), Air Borne Force (ABF), Army Recruit Training School (ARTS), 1st Battalion Infantry (1 BN) and 5th Battalion Infantry (5 BN)]. Tick sampling was conducted in three ecological zones namely; Coastal, Guinea and Sudan savanna between June 2016 and December 2017. The map of Ghana with the geographical regions and study sites was created using ArcGIS® software by Esri (www. esri. com) N imo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 5 of 11 pooled infection status with animal host and ecological Table 1 Background information of ticks collected zone was determined using Pearson Chi-square. Statisti- Total Coastal savanna Guinea cal significance was set at a P-value < 0.005. n (%) n (%) and Sudan savanna n (%) Results Animal host Species composition of ticks Cattle 1674 (83.0) 728 (98.5) 946 (74.1) A total of 2016 ticks were collected, of which 66 and Dogs 325 (16.1) 11 (1.5) 314 (24.6) 34% were males and females, respectively. The majority Goats 7 (0.3) 0 (0.0) 7 (0.5) of ticks sampled (63.3%) were from Guinea and Sudan Sheep 10 (0.5) 0 (0.0) 10 (0.8) savanna, while the remaining ticks came from the Coastal Sex of ticks savanna of Ghana (Table  1). Seven tick species were Male 1327 (65.8) 345 (46.7) 982 (76.9) identified, with Amblyomma variegatum (60%) being Female 689 (34.2) 394 (53.3) 295 (23.1) the most abundant (Fig.  2). The majority of the A. var- iegatum were from cattle (99.8%), whereas Rhipicephalus sanguineus sensu lato (s.l.) were found more frequently Out of 912 tick pools tested, the overall infection in dogs (73%) (see Additional file 1). Rhipicephalus (Boo- rates for C. burnetii and Rickettsia spp. were 7.9% (95% philus) sp. (0.1%) was only found in the Coastal savanna. CI 6.8–9.2) and 24.5% (95% CI 22.4–26.6), respectively. Generally, Rhipicephalus species occurred more fre- Amblyomma variegatum, the most prevalent species in quently in the Coastal savanna (34%) than in the Guinea the tick pools, recorded infection rates of 11.0% (95% CI and Sudan savanna (31%). Significant differences were 9.3–13.0) and 38.6% (95% CI 38.3–42.1) for C. burnetii recorded in the distribution of R. sanguineus s.l. ( 2χ = and Rickettsia spp., respectively (Table 3). 161.32, df = 1, P < 0.0001), Rhipicephalus spp. ( 2χ = Overall co-infection with both C. burnetii and Rick- 387.57, df = 1, P < 0.0001), Hyalomma truncatum ( 2χ = ettsia spp. was identified to be 9.6% (95% CI 8.0–11.7) 75.59, df = 1, P < 0.0001) and Hyalomma rufipes (χ 2 = (Table 4). All co-infections observed were in cattle, with 36.93, df = 1, P < 0.0001) from the three ecological zones approximately 7.6% being true co-infection in single sam- of Ghana (Fig. 3). ple pools. No co-infections were recorded at one site (1st Battalion Infantry) in the Coastal savannah or Navrongo Pathogen detection and identification of tick pools in the Sudan savanna. The majority of co-infections were Coxiella burnetii was detected at all sites except Nav- identified in the Guinea 41% (39/96) and Coastal savanna rongo, with a pooled positive rate of 16.7%, whereas a 31% (30/96) ecological zones. Two of the sites, one in the pooled positive rate of 45.6% was recorded for Rickettsia Coastal and the other in Guinea savanna, recorded co- spp. and was detected at all seven sites (Table 3). Coxiella infections of 6% (n = 6) and 1% (n = 1), respectively. burnetii was only detected in ticks collected from cattle; however, Rickettsia spp. were identified in cattle, goats Discussion and dogs. The number of tick pools positive for C. bur- Studies on tick distribution, population and disease pres- netii and Rickettsia spp. was significant with respect to ence are sparse in Ghana and are needed to better under- the animal host and study sites of tick sampling (Table 2). stand the risk of tick-borne infections within Ghana’s CCHFV and AHFV were not detected in any of the tick various ecologic zones. The ticks collected in this study pools. reinforce previously recorded distributions of tick spe- cies in Ghana [36]. Of the ticks collected, several are associated with the transmission of human pathogens. Spatial distribution and infection rates Amblyomma variegatum is widely distributed through- Rickettsia spp. were identified at all seven study sites, out Ghana and has been collected from both domestic with Guinea and Sudan savanna ecological zones record- and wild animals [36–38]. Therefore, it is predictable that ing 66% (273/416) of the rickettsial infections in the tick in this study, A. variegatum was the most abundant tick pools (Table 2). While C. burnetii was detected in simi- species collected. Generally, in the ecozones studied, A. lar pool numbers in the Coastal (77/152) and Guinea variegatum seemed to thrive well. This distribution and (75/152) ecological zones, no infected ticks were identi- abundance are of note, however, as A. variegatum has fied at the Navrongo site situated in the Sudan savanna been implicated as a vector of diseases such as ATBF and ecological zone. CCHF [39, 40]. The Hyalomma spp. are known vectors Nimo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 6 of 11 Fig. 2 Overall distribution of tick species identified from the three ecological zones. Domestic animals (cattle, goats, dogs) were examined for ticks between June 2016 and December 2017 for CCHFV, and R. sanguineus s.l. has been implicated were from communities in which the livestock were pre- as a vector for some Rickettsia spp. including those in sent and normally accompany the domestic ruminants the spotted fever group [4, 5, 39]. Currently, ATBF has to grazing fields. It is unclear whether this is a host pref- not been recorded in Ghana; however many neighbour- erence or simply that the domestic dogs are within the ing countries have reported cases. The presence of Hya- same habitat as the livestock, and therefore the ticks were lomma spp. and R. sanguineus s.l. at the study sites could feeding opportunistically. support the transmission of the pathogens that cause Coxiella burnetii (16.7%) and Rickettsia spp. (45.6%) CCHFV and rickettsioses. Nonetheless, CCHFV was not were detected in the pooled tick samples collected in this identified in any of the Hyalomma spp. and this could be study. For both C. burnetii and Rickettsia spp., the infec- due to low sample size or that they are not vectors of the tion rate was highest in the tick species that are often disease in Ghana. associated with the transmission of Q fever and rickett- Previous studies have found that tick species, includ- sial diseases, respectively, including A. variegatum, Rhi- ing H. rufipes, H. truncatum and R. evertsi, are found picephalus spp., H. truncatum and H. rufipes [7, 39–41]. almost exclusively on domestic ruminants [36–38]. This Coxiella burnetii is often asymptomatic in cattle, barring observation could be explained by host preference and/ some reproductive issues, and causes chronic infections or environmental preference of these tick species as some [16]. Studies have reported that C. burnetii is at par- studies have found that these factors impact tick distribu- ticularly high concentrations during times of parturi- tion in Ghana [37]. Ticks taken from dogs in this study tion in infected animals, possibly increasing the risk of N imo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 7 of 11 Fig. 3 Distribution of tick species identified morphologically from livestock by ecological zone between June 2016 and December 2017 using χ2 test transmission to anyone handling or in contact with new- co-infection in single sample pools accounted for 7.6%. borns, placental tissues and amniotic fluids [42]. Cases of Co-infection of pathogens within ticks is fairly common Q fever have been reported in Ghana, including an out- [45–47]. Furthermore, co-infection of C. burnetii and break in the Volta region associated with interaction with Rickettsia spp. has been recorded in several tick species small ruminants [20, 22]. and is often the most common combination found [4, In the present study, approximately 25% of the ticks 45, 48]. These co-infections may be the result of sub- tested were infected with Rickettsia spp. The highest rate sequent blood meals, feeding on a co-infected host or of infection was in A. variegatum, possibly due to the co-feeding with other infected ticks [4, 49]. Multiple wide distribution of A. variegatum throughout Ghana. pathogen co-infection of ticks may lead to many com- A few cases of rickettsial diseases have been reported plications, including further spread and distribution of in Ghana. Past studies have documented the detection diseases and complications of clinical diagnosis. Mul- of Rickettsia spp. in ticks and humans in Ghana, though tiple tick pathogen co-infection in humans may com- they were not further identified to species [43, 44]. It is plicate clinical diagnosis and, subsequently, proper possible that R. africae or other Rickettsia spp. pathogens treatment [4, 50]. Pathogens may require different are present in Ghana, which highlights the importance treatments; therefore, mis- or undiagnosed co-infec- of further research and identification of all Rickettsia tions may result in prolonged illness in patients [4, 50]. spp. detected. Additionally, since livestock are allowed to Tick collection was not conducted periodically roam freely, there are increased opportunities for ticks to throughout the year, and as a result, the study did not be distributed to novel areas of Ghana and consequently associate tick distribution with the dry and wet sea- potential transmission of tick-borne pathogens such as R. sons of Ghana. The study is also limited in selecting africae to humans. sampling sites and, therefore, could not cover all the Four of the seven tick species were positive for C. Ghanaian ecological zones to provide a better repre- burnetii and Rickettsia spp. co-infection. Additionally, sentation of tick and tick-borne pathogens in the entire the Rhipicephalus spp. pools had the highest rate of co- country. Another limitation of this study is that the infection with these two pathogens. However, actual majority of the co-infections detected were in pooled Nimo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 8 of 11 Table 2 Association between infection rate of detected pathogens in ticks collected from different hosts and sampling sites using χ2 test Pooled positive, n (%) Pooled positive, n (%) Total pooled n (%) Coxiella burnetii χ2 df P‑value Rickettsia spp. χ2 df P‑value Mammalian host 35.61 3 < 0.001 141.63 3 < 0.001 Cattle 763 152 (19.9) 414 (54.3) Dogs 141 0 (0.0) 1 (25.0) Sheep 4 0 (0.0) 0 (0.0) Goats 4 0 (0.0) 1 (0.7) Study site 9.35 1 0.002 7.89 1 0.005 Coastal savanna 168 66 (39.3) 44 (26.2) 5 BN 103 10 (9.7) 64 (62.1) ART 89 1 (1.1) 35 (39.3) 1 BN Guinea and Sudan savanna 6 BN 99 50 (50.5) 79 (79.8) AF 127 23 (18.1) 83 (65.4) Navrongo 174 0 (0.0) 5 (2.9) ABF 152 2 (1.3) 106 (69.7) N imo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 9 of 11 Table 3 Infection rate of tick pools with Coxiella burnetii and Rickettsia species Coxiella burnetii Rickettsia spp. No. of Min. pool size Max pool size No. of Mean No. of Mean infection rate (95% pools positive infection rate positive CI) tested pools (95% CI) pools Amblyomma variegatum 516 1 5 123 11.0 (9.3–13.0) 344 38.6 (38.3–42.1) Rhipicephalus sanguineus 200 1 3 3 0.7 (0.2–2.2) 15 3.5 (2.1–5.8) s.l. Rhipicephalus spp. 99 1 5 19 10.5 (6.8–16.0) 22 12.2 (8.2–18.0) Hyalomma truncatum 56 1 3 5 4.5 (1.9–70.5) 19 17.0 (11.0–25.7) Hyalomma rufipes 30 1 3 2 3.8 (1.0–14.5) 14 30.2 (18.8–46.2) Rhipicephalus evertsi 10 1 3 0 0 1 5.1 (0.7–31.4) Rhipicephalus (Boophilus) 1 1 1 0 0 1 100 sp. Total 912 1 5 152 7.9 (6.8–9.2) 416 24.5 (22.4–26.6) Table 4 Co‑infection of pooled tick species Coxiella burnetii and Rickettsia spp. Min. pool size Max pool size No. of co‑infection pool Mean infection rate (95% CI) Amblyomma variegatum 1 3 82 9.6 (7.8–11.8) Rhipicephalus sanguineus s.l. 1 3 0 0 Rhipicephalus spp. 1 2 10 19.1 (10.7–32.5) Hyalomma truncatum 1 3 3 9.1 (3.0–25.7) Hyalomma rufipes 1 3 1 3.6 (0.5–23.2) Rhipicephalus evertsi 1 3 0 0 Rhipicephalus (Boophilus) sp. 1 1 0 0 Total 1 3 96 9.6 (8.0–11.7) samples of two or three ticks, and therefore, one can- Abbreviation not differentiate whether these are actual co-infections CI: Confidence interval. or whether multiple ticks within a pooled sample each carried one pathogen. Supplementary Information The online version contains supplementary material available at https:// doi. org/1 0.1 186/s 13071‑0 22‑0 5208‑8. Conclusions Additional file 1. Distribution of tick species collected from livestock dur‑ Among approximately 2000 ticks mostly obtained from ing the study period. cattle that were examined from three ecologic zones in Ghana, 2016 ticks were identified, with 16.7% and 45.6% Acknowledgements rates of Coxiella and Rickettsia species, respectively. As We appreciate the invaluable support of the Ghana Armed Forces Veterinary the world becomes more globalized and trade between Department, the Vector Team at the Navrongo Health Research Centre and the Noguchi Memorial Institute for Medical Research. The United States countries increases, it is imperative that research on Army Medical Research Institute of Infectious Diseases (USAMRIID) provided current tick species and possible disease introduction assistance in the molecular assays and we the authors are very grateful. Many to new areas be monitored. Continued surveillance and thanks to Dr. James Harwood for proofreading and reviewing the manuscript. pathogen testing are important to track the possible Authors’ contributions introduction of new tick pathogens entering the coun- SCNP, JWD and AL conceived and designed the study; MM, SOA, REB, DL, JT, try. As the importation of livestock increases to meet CA, CD and VA realized the fieldwork; MM, SOA, REB, JT, DL and JWK performed molecular testing; EB and AL did the data analysis; MM, SOA, EB and LT wrote the demand for dairy and meat products, so does the the first draft of the manuscript; SCNP, AF, CW and RJS revised the manuscript; likelihood for the importation of new diseases. SD, AL and JWD supervised the study. All authors read and approved the final manuscript. Nimo‑Paintsil et al. Parasites & Vectors (2022) 15:86 Page 10 of 11 Authors’ disclaimer statement emerging countries. Exp Appl Acarol. 2011;54:65–83. https:// doi. org/ 10. The views expressed in this article are those of the authors and do not neces‑ 1007/s 10493‑0 10‑ 9414‑4. sarily reflect the official policy or position of the Department of the Navy, 4. Reye AL, Arinola OG, Hubschen JM, Muller CP. Pathogen prevalence in Department of Defense, or the US Government. Opinions, interpretations, ticks collected from the vegetation and livestock in Nigeria. Appl Environ conclusions, and recommendations are those of the authors and are not Microbiol. 2012;78:2562–8. https:// doi. org/ 10. 1128/ AEM.0 6686‑ 11. necessarily endorsed by the US Army. 5. Telmadarraiy Z, Chinikar S, Vatandoost H, Faghihi F, Hosseini‑Chegeni A. Vectors of Crimean Congo hemorrhagic fever virus in Iran. J Arthropod‑ Copyright assignment statement Borne Dis. 2015;9:137–47. Dr. Nimo‑Paintsil, LT Arimoto, LT Lura, LT Watters, LCDR Diclaro II, and CDR 6. Estrada‑Pena A, de la Fuente J. The ecology of ticks and epidemiology of Letizia are military service members or employees of the US Government. This tick‑borne viral diseases. Antivir Res. 2014;108:104–28. https://d oi. org/ 10. work was prepared as part of their official duties. Title 17 USC §105 provides 1016/j. antivi ral. 2014. 05. 016. that “Copyright protection under this title is not available for any work of 7. Bente DA, Forrester NL, Watts DM, McAuley AJ, Whitehouse CA, Bray M. the United States Government”. Title 17 USC §101 defines a US Government Crimean‑Congo hemorrhagic fever: history, epidemiology, pathogenesis, work as a work prepared by a military service member or employee of the US clinical syndrome and genetic diversity. Antivir Res. 2013;100:159–89. Government as part of that person’s official duties. https://d oi. org/ 10. 1016/j.a ntivi ral.2 013. 07. 006. 8. Hawman DW, Feldmann H. Recent advances in understanding Crimean‑ Funding Congo hemorrhagic fever virus. 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Amoako N, Duodu S, Dennis FE, Bonney JHK, Asante KP, Ameh J, et al. • thorough peer review by experienced rese archers in your field Detection of dengue virus among children with suspected malaria, Accra, • rapid publication on acceptance Ghana. Emerg Infect Dis. 2018;24:1544–7. https:// doi. org/ 10. 3201/ eid24 • support for research data, including large and complex data types 08. 180341. 45. Nooroong P, Trinachartvanit W, Baimai V, Ahantarig A. Phylogenetic stud‑ • gold Open Access which fosters wider collaboration and increased citations ies of bacteria (Rickettsia, Coxiella, and Anaplasma) in Amblyomma and • maximum visibility for your research: over 100M website views per year Dermacentor ticks in Thailand and their co‑infection. Ticks Tick‑Borne Dis. 2018;9:963–71. https:// doi. org/ 10. 1016/j.t tbdis. 2018. 03. 027. At BMC, research is always in progress. 46. Raileanu C, Moutailler S, Pavel I, Porea D, Mihalca AD, Savuta G, et al. 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