Molecular Detection Of Tick-borne Pathogens Research Articles

Molecular Detection of Tick-Borne Pathogens in Dogs from Indigenous Communities, Amazon, Brazil.

Background: There are few reports of tick-borne pathogens infecting dogs living in indigenous communities of Brazil. Herein, we aimed to molecularly detect vector-borne pathogens in dogs from two indigenous communities in the Brazilian Amazon. Materials and Methods: We surveyed 327 dogs raised in Amazon region at 2 distinct indigenous ethnicities for the molecular detection of tick-borne pathogens (114 from Tapirapé and 213 from Karajá indigenous ethnicity). Whole blood samples were subjected to PCR and sequencing for Ehrlichia, Babesia, and Hepatozoon. Univariate and multivariate analyses were used to investigate the factors affecting the pathogen infection patterns in dogs. Results: Among the 327 blood samples, 40 were positive for Ehrlichia canis (12.2%), 2 for Anaplasma platys (0.61%), and 204 were positive for Hepatozoon canis (66.5%). Binary Logistic Regression showed association between E. canis infection and ethnicity (p = 0.010) and tick attachment (p = 0.041). Karajá dogs were 3.4 times (95% CI 1.3-8.5) more likely to be positive for E. canis than Tapirapé dogs. Dogs with ticks were 2.5 times more likely (95% CI 1.0-7.6) to be positive for E. canis than dogs without ticks. Conclusions: Our survey expands the knowledge regarding the presence of vector-borne pathogens in dogs from indigenous communities in the Amazon region.

Molecular Detection of Anaplasma phagocytophilum in Ixodid Ticks Infesting White Fulani Cattle in Zaria and its Environs, Kaduna State, Nigeria.

Anaplasmosis, a disease caused by various species of Anaplasma, poses important economic constraints to animal breeders. In Zaria, molecular detection of tick-borne pathogens infecting cattle is very sketchy. Being hematophagous, ticks are capable of transmitting disease agents such as viruses, bacteria and protozoa. This study was carried out to detect for the presence of Anaplasma pathogen in the various tick species infesting white Fulani breeds of cattle owing to their high population in Zaria and environs using a Polymerase Chain Reaction (PCR). Three hundred and eighty-four (n=384) white Fulani breed of cattle of varying age and sex were sampled from 32 herds in 4 villages (Bomo, Tofu, Ungwan Dabosa and Majeru). The DNA was extracted using Qiagen commercial kit following manufacturer’s instructions. Genomic DNA of the species of ticks was amplified in a semi-nested PCR targeting the 16S rRNA fragment of the Anaplasma spp at expected amplicon size of 711bp. The PCR products were purified using Gel Extraction Kit (Bioland, Scientific LLC) following the manufacturer’s protocol. All sequences were subjected to a Basic Local Alignment Search Tool (BLAST) to determined their identities and assess their homologues and similarities to those in the GenBank. A neighbor-joining tree was generated using the Molecular Engineering Genetic Analysis (MEGA 7.0) to derive the close relationship between the isolates. The result of this study implies that Anaplasma spp (MN044909) is present in all the five species of ticks infesting cattle in Zaria, Kaduna State, Nigeria and pose a high risk on humans and animals health.
 Anaplasmosis, a disease caused by various species of Anaplasma, poses important economic constraints to animal breeders. In Zaria, molecular detection of tick-borne pathogens infecting cattle is very sketchy. Being hematophagous, ticks are capable of transmitting disease agents such as viruses, bacteria and protozoa. This study was carried out to detect for the presence of Anaplasma pathogen in the various tick species infesting white Fulani breeds of cattle owing to their high population in Zaria and environs using a Polymerase Chain Reaction (PCR). Three hundred and eighty-four (n=384) white Fulani breed of cattle of varying age and sex were sampled from 32 herds in 4 villages (Bomo, Tofu, Ungwan Dabosa and Majeru). The DNA was extracted using Qiagen commercial kit following manufacturer’s instructions. Genomic DNA of the species of ticks was amplified in a semi-nested PCR targeting the 16S rRNA fragment of the Anaplasma spp at expected amplicon size of 711bp. The PCR products were purified using Gel Extraction Kit (Bioland, Scientific LLC) following the manufacturer’s protocol. All sequences were subjected to a Basic Local Alignment Search Tool (BLAST) to determined their identities and assess their homologues and similarities to those in the GenBank. A neighbor-joining tree was generated using the Molecular Engineering Genetic Analysis (MEGA 7.0) to derive the close relationship between the isolates. The result of this study implies that Anaplasma spp (MN044909) is present in all the five species of ticks infesting cattle in Zaria, Kaduna State, Nigeria and pose a high risk on humans and animals health.

Detection of Ticks and Tick-Borne Pathogens of Urban Stray Dogs in South Africa.

This study aimed to identify ticks infesting dogs admitted to the Potchefstroom Animal Welfare Society (PAWS) and to detect tick-borne pathogens they are harbouring. A total of 592 ticks were collected from 61 stray dogs admitted to PAWS originating from several suburbs in and near Potchefstroom, South Africa. The dog ticks were identified as Haemaphysalis elliptica (39%) and Rhipicephalus sanguineus (61%) by both morphological and DNA analyses. Of these ticks, H. elliptica consisted of 67.5% (156/231) and 32.5% (75/231) female and male ticks, respectively, whilst R. sanguineus consisted of 48.5% (175/361) and 51.5% (186/361) female and male ticks, respectively. Microscopic examination of blood smears from engorged female ticks indicated overall occurrences of 0.5% (1/204) for Babesia spp. from R. sanguineus, 1% (2/204) of Anaplasma spp. from H. elliptica, and 22% (45/204) of Rickettsia spp. from both H. elliptica and R. sanguineus. Using pooled samples molecular detection of tick-borne pathogens indicated overall occurrences of 1% (1/104) for A. phagocytophilum in H. elliptica, 9.6% (10/104) of Rickettsia spp. in H. elliptica and R. sanguineus, 5.8% (6/104) of Ehrlichia canis in H. elliptica and R. sanguineus, and 13.5% (14/104) of Coxiella spp. in both H. elliptica and R. sanguineus. Additionally, PCR detected 6.5% (2/31) of Coxiella spp. DNA from H. elliptica eggs. Our data indicate that urban stray dogs admitted at PAWS are infested by H. elliptica and R. sanguineus ticks which are harbouring several pathogenic organisms known to cause tick-borne diseases.

Molecular detection of tick-borne pathogens in Rhipicephalus sanguineus sensu stricto collected from dogs in the steppe and high plateau regions of Algeria

Epidemiology and distributions of canine tick-borne diseases as well as their veterinary and zoonotic significance are poorly understood in Algeria. The present study describes a molecular investigation of important tick-borne pathogens in Rhipicephalus sanguineus sensu stricto collected from domestic dogs in steppe and high plateau areas of central and eastern Algeria. In total, 1,043 ticks were collected from 147 dogs, including 756 ticks from 124 dogs in the steppe region of Djelfa and 287 ticks from 23 dogs in the high plateau area of Bordj Bou Arreridj. Ticks were divided into 384 pools (309 pools from Djelfa and 75 pools from Bordj Bou Arreridj) and tested for genomic materials of Crimean-Congo hemorrhagic fever virus (CCHFV) as well as DNA for Anaplasma phagocytophilum, Anaplasma platys, Ehrlichia canis, Rickettsia spp., Babesia spp., and Hepatozoon spp. using PCR, sequencing and phylogenetic analysis. Hepatozoon spp. was most prevalent, with 160 positive pools (41.7%), and 12 of these were sequenced and identified as Hepatozoon canis. Babesia spp. was detected in 50 samples (13.0%), of which 11 were sequenced and identified as Babesia vogeli. A. platys and E. canis were detected in 92 (24.0%) and 15 (3.9%) of tested samples, respectively. Rickettsia spp. were detected in 24 (6.3%) samples, including 11 samples identified as R. massiliae, 6 samples identified as R. conorii conorii, and 7 samples could not be identified to species level. All 384 pools tested negative for CCHFV and A. phagocytophilum. In addition to detection of R. conorii conorii, R. massiliae, and E. canis, the present study provides the first molecular data for occurrence of A. platys, B. vogeli, and H. canis in Rh. sanguineus s.s. infesting dogs in Algeria. Further large scale studies should be conducted to better understand the epidemiology, distributions, and importance of canine tick-borne pathogens in Algeria.

Prevalence of Ehrlichia-, Babesia-, and Hepatozoon-infected brown dog ticks in Khon Kaen Province, Northeast Thailand.

Background and Aim:The brown dog tick, Rhipicephalus sanguineus sensu lato, is the most common tick found on domestic dogs in Southeast Asia, including Thailand. Canine tick-borne pathogens are a public health concern worldwide. Tick-borne diseases are diagnosed by identifying pathogens based on the morphological or molecular analyses of dog blood samples. However, the collection of ticks, a non-invasive procedure, is easier than drawing blood. This study aimed to demonstrate the usefulness of collecting brown dog ticks for the diagnosis of tick-borne diseases and for estimating the prevalence of tick-borne pathogens among companion dogs in Khon Kaen, Northeast Thailand.Materials and Methods:Seventy brown dog ticks from 70 companion dogs in Khon Kaen Province, Thailand, were evaluated for molecular evidence of tick-borne pathogens, including Babesia spp., Ehrlichia canis, and Hepatozoon canis. Ticks were collected from dogs at a private animal hospital based on the presence of at least one of the three inclusion criteria: fever, anorexia, or lethargy. Molecular diagnosis was performed using conventional polymerase chain reaction for the detection of pathogens.Results:Of the 70 ticks collected from 70 sick dogs, 55 (78.57%) were positive for tick-borne pathogens. The most common infection was a single infection with H. canis (65.71%) followed by Babesia spp. (31.43%) and E. canis (30.00%). Coinfection was observed in 14 ticks (20.00%), and coinfection with Babesia spp. and E. canis was the most prevalent double infection (n = 6). The prevalence of coinfection was identical for H. canis mixed with Babesia spp. and H. canis mixed with E. canis (n = 4).Conclusion:The present study showed that tick-borne pathogens are highly prevalent among companion dogs in Khon Kaen Province. Therefore, we encourage an increase in tick control or the reduction and prevention of tick-borne diseases in this region. Furthermore, this study revealed that ticks are valuable samples for the molecular detection of tick-borne pathogens.

No molecular detection of tick-borne pathogens in the blood of patients with erythema migrans in Belgium

BackgroundA number of tick-borne pathogens circulate in the Belgian tick population in addition to the causative agent of Lyme borreliosis. However, so far, only a few patients with tick-borne diseases other than Lyme borreliosis have been reported in Belgium. The aim of this study was to investigate the occurrence of other human tick-borne infections in Belgium and their possible clinical manifestation.MethodsPatients with fever (> 37.5 °C) after a tick bite or those with erythema migrans (EM) were included in the study. EDTA-blood samples were screened for the presence of DNA from Borrelia burgdorferi sensu lato, Borrelia miyamotoi, Anaplasma phagocytophilum, Neoehrlichia mikurensis, spotted fever group rickettsiae (genus Rickettsia), Babesia spp., Bartonella spp., Spiroplasma ixodetis and tick-borne encephalitis virus, using multiplex PCR methods. A questionnaire on, among others, demographics and clinical symptoms, was also filled in.ResultsOver a period of 3 years, 119 patients with EM and 14 patients with fever after a recent tick bite were enrolled in the study. Three samples initially tested positive for N. mikurensis by quantitative PCR (qPCR), but the results could not be confirmed by other PCR methods, and repetition of the DNA extraction procedure and qPCR test was not successful. The qPCR test results for the other tick-borne pathogens were negative.ConclusionsIn general, only a few patients with fever after a tick bite could be identified. Although no tick-borne pathogens were detected, their occurrence cannot be excluded based on the limited number of patients and the limitations inherent to current methodologies. This study underscores the possibility of false-positive PCR results and the necessity for the development of multiple independent tools for the sensitive and specific detection of emerging tick-borne pathogens.Graphical

Molecular Detection of Tick-Borne Pathogens in American Bison (Bison bison) at El Uno Ecological Reserve, Janos, Chihuahua, Mexico.

American bison (Bison bison) is listed as near-threatened and in danger of extinction in Mexico. Recent studies have demonstrated the presence of several emerging pathogens at the Janos Biosphere Reserve (JBR), inhabited by one wild herd of American bison. Blood samples were collected from 26 American bison in the JBR. We tested for the presence of Anaplasma marginale, Babesia bigemina, B. bovis, Borrelia burgdorferi sensu lato, and Rickettsia rickettsii DNA using nested and semi-nested PCR protocols performing duplicates in two different laboratories. Results showed three animals (11.5%) positive for B. burgdorferi s. l., three more (11.5%) for Rickettsia rickettsii, and four (19.2%) for B. bovis. Two individuals were co-infected with B. burgdorferi s. l. and B. bovis. We found no animals positive for A. marginale and B. bigemina. This is the first report in America of R. rickettsii in American bison. American bison has been described as an important reservoir for pathogens of zoonotic and veterinary importance; thus, the presence of tick-borne pathogen DNA in the JBR American bison indicates the importance of continuous wildlife health surveys.

Molecular detection of tick-borne pathogens harbored by ticks collected from local breed (Non-descript) of cattle in the chhattishgarh state of India

Molecular detection of tick-borne pathogens harbored by ticks collected from local breed (Non-descript) of cattle in the chhattishgarh state of India

Molecular detection of tick-borne pathogens in infected dogs associated with Rhipicephalus sanguineus tick infestation in Thailand.

Background and Aim:Tick-borne pathogens (TBPs) are of great concern having the potential to threaten canine health. Dogs infected with Ehrlichia canis, Anaplasma platys, Babesia canis, and Hepatozoon canis are commonly found in Thailand; Rhipicephalus sanguineus tick is the most common vector of diseases. This study aimed to determine the prevalence of common TBPs in dogs and their ticks in Thailand using polymerase chain reaction (PCR) and DNA sequencing methods.Materials and Methods:Forty-four blood samples were positively diagnosed with TBPs infection by microscopy. Samples were from animal hospitals in Maha Sarakham, Amnat Charoen, Nakhon Ratchasima, and Bangkok, Thailand, during January-June 2020. Five to six ticks were also taken from infected dogs, and then, both blood and tick were analyzed using PCR and DNA sequencing.Results:PCR results showed that R. sanguineus was the only tick species detected in this study. The appearance of single infection with E. canis was the most common infection found in dogs and ticks (64% and 82%, respectively). Correlation of pathogen infection in hosts and their vector was performed by similarity detection of pathogens between blood and tick samples based on PCR analysis in 29 samples (66%) but there was no significant differentiation.Conclusion:E. canis appears as the most common canine tick-borne pathogen in Thailand, which was detected in both healthy and sick dogs as well as in R. sanguineus. The findings show the relationships among host dogs, pathogens, and ticks. Veterinarians should be proactive in educating pet owners about the risks associated with ticks and their important pathogens and plan effective control strategies.

Tick species identification and molecular detection of tick-borne pathogens in blood and ticks collected from cattle in Egypt

To address the lack of information on ticks infesting cattle in Egypt and the pathogens that they transmit, the current study aimed to (i) provide insight into tick species found on cattle in Egypt, (ii) identify the pathogens in ticks and their cattle hosts and (iii) detect pathogen associations in ticks and cattle. Tick samples and blood from their bovine hosts were collected from three different areas in Egypt (EL-Faiyum Oasis, Assiut Governorate and EL-Kharga Oasis). Tick species were identified by morphology and by sequence analysis of the cytochrome C oxidase subunit 1 (cox1) gene. Tick pools and blood samples from cattle were screened by the Reverse Line Blot hybridization (RLB) assay for the simultaneous detection of tick-borne pathogens, including Babesia, Theileria, Anaplasma, Ehrlichia, and Rickettsia spp., as well as the tick endosymbiont Midichloria mitochondrii. The RLB results were confirmed with specific conventional and semi-nested PCRs followed by sequencing. In total, 570 ticks (males, females and nymphs) were collected from 41 heads of cattle. Altogether 398 ticks belonged to the genus Hyalomma (397 Hyalomma excavatum and one Hyalomma scupense) while 172 ticks were identified as Rhipicephalus annulatus. Pooled H. excavatum ticks tested positive for several protozoa and bacteria with different minimum infection rates (MIRs): Theileria annulata (18.1 %), Babesia occultans (1.8 %), Anaplasma marginale (28.5 %), Anaplasma platys (0.25 %), Midichloria mitochondrii (11.6 %), Ehrlichia chaffeensis-like (1.8 %) and Ehrlichia minasensis (1 %). In R. annulatus, several agents were identified at different MIRs: T. annulata (2.3 %), B. bovis (0.6 %), A. marginale (18.0 %), A. platys (1.2 %), M. mitochondrii (2.9 %), E. minasensis (0.6 %). Pathogens co-detection in tick pools revealed A. marginale and T. annulata in 13.3 % samples followed by the co-detection of A. marginale and M. mitochondrii (8.4 %). In addition, triple co-detection with A. marginale, T. annulata and M. mitochondrii were found in 5.3 % of the tick pools. In cattle, the most common coinfection was with A. marginale and T. annulata (82.9 %) followed by the coinfection between A. marginale, T. annulata and B. bovis (4.9 %), A. marginale and B. bigemina (2.4 %) and finally the coinfection between T. annulata and B. occultans (2.4 %). Anaplasma platys, Babesia occultans, and E. minasensis were detected for the first time in Egypt in both cattle and ticks. These findings should be taken in consideration regarding human and animal wellbeing by the public health and veterinary authorities in Egypt.

Morphological identification of ticks and molecular detection of tick-borne pathogens from bare-nosed wombats (Vombatus ursinus)

BackgroundTicks are obligate haematophagous ectoparasites of vertebrate hosts and transmit the widest range of pathogenic organisms of any arthropod vector. Seven tick species are known to feed on bare-nosed wombats (Vombatus ursinus), in addition to the highly prevalent Sarcoptes scabiei mite which causes fatal sarcoptic mange in most bare-nosed wombat populations. Little is known about the pathogens carried by most wombat ticks or how they may impact wombats and wombat handlers.MethodsWombat ticks were sourced from wildlife hospitals and sanctuaries across Australia and identified to species level using taxonomic keys. Genomic DNA was extracted from a subsample, and following the amplification of the bacterial 16S rRNA gene V3–V4 hypervariable region, next-generation sequencing (NGS) on the Illumina MiSeq platform was used to assess the microbial composition.ResultsA total of 447 tick specimens were collected from 47 bare-nosed wombats between January 2019 and January 2020. Five species of ticks were identified comprising wombat tick Bothriocroton auruginans (n = 420), wallaby tick Haemaphysalis bancrofti (n = 8), bush tick Haemaphysalis longicornis (n = 3), common marsupial tick Ixodes tasmani (n = 12), and Australian paralysis tick Ixodes holocyclus (n = 4). Tick infestations ranged from one to 73 ticks per wombat. The wombat tick was the most prevalent tick species comprising 94% of the total number of samples and was present on 97.9% (46/47) of wombat hosts. NGS results revealed the 16S rRNA gene diversity profile was predominantly Proteobacteria (55.1%) followed by Firmicutes (21.9%) and Actinobacteria (18.4%). A species of Coxiella sharing closest sequence identity to Coxiella burnetii (99.07%), was detected in 72% of B. auruginans and a Rickettsiella endosymbiont dominated the bacterial profile for I. tasmani.ConclusionsA new host record for H. longicornis is the bare-nosed wombat. One adult male and two engorged adult female specimens were found on an adult male wombat from Coolagolite in New South Wales, and more specimens should be collected to confirm this host record. The most prevalent tick found on bare-nosed wombats was B. auruginans, confirming previous records. Analysis of alpha-diversity showed high variability across both sample locations and instars, similar to previous studies. The detection of various Proteobacteria in this study highlights the high bacterial diversity in native Australian ticks.Graphical

A report on tick burden and molecular detection of tick-borne pathogens in cattle blood samples collected from four regions in Saudi Arabia

Babesiosis, theileriosis and anaplasmosis are among the most commonly reported tick-borne diseases in cattle and are associated with significant economic losses. Through the present study the researchers aimed to report the presence of various pathogens that cause babesiosis, theileriosis and anaplasmosis in cattle collected from different provinces in Saudi Arabia and to report their phylogenetic relationship. A total of 362 blood samples of cattle along with ticks that were present on the cattle were collected from four regions (Riyadh, Al-Kharj, Al-Hasa and Al-Qassim) of Saudi Arabia. Blood samples were screened by polymerase chain reaction (PCR) for the presence of various Babesia, Theileria and Anaplasma species by amplification of their 18S rRNA and/or 23S rRNA genes. A total of 541 ticks were collected and identified from the cattle. These included Hyalomma anatolicum, Hyalomma dromedarii, Hyalomma impeltatum, Hyalomma excavatum, Rhipicephalus annulatus and Rhipicephalus turanicus. Regarding tick-borne pathogens, the overall prevalence was 1.9 % (7/362) for Theileria annulata, (2/362) 0.6 % for Theileria and (21/362) 5.8 % for Anaplasma ovis. Four of the cattle were found to be co-infected with more than one pathogen (1.1 %). We did not detect any Babesia species in the blood of the studied cattle. Prevalence of the Theileria and Anaplasma species was highest in cattle that resided in Riyadh, followed by cattle from Al-Hasa and Al-Qassim. Representative amplified partial-gene sequences of T. annulata (GenBank accession numbers MK826137-39) and A. ovis (GenBank acc. no. MK 880224) were submitted to GenBank. The presence of ticks on cattle was found to be associated with a high prevalence of Theileria spp. (P = 0.02) and Anaplasma ovis (P < 0.001). We report novel genotypes of T. annulata and A. ovis from cattle in Saudi Arabia and we recommend that molecular surveys are undertaken throughout the country to address the prevalence and geographical distribution of tick-borne infections for their effective diagnosis and treatment.

Molecular detection of tick-borne pathogens in ticks collected from pets in selected mountainous areas of Tatra County (Tatra Mountains, Poland)

The mountainous and foothill areas, in which the city of Zakopane, the capital of Tatra County, is located are characterized by continuous weather changes, lower air temperature, persistent snow cover, and poorer vegetation than in the lowlands. Ixodes ricinus and Ixodes hexagonus are vectors of tick-borne diseases and play an important role in the persistence of tick-borne diseases. The aim of the study was to determine the risk of exposure of domestic cats and dogs to the attacks of Ixodid ticks, to tick-borne infections with Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum, Babesia microti and Toxoplasma gondii in the city of Zakopane and the surrounding area. In 2017–2018 ticks were collected from a total of 10 domestic cats and 88 domestic dogs. Selected pathogens of tick-borne diseases were detected by PCR and nested PCR. The study material contained 119 I. ricinus and 36 I. hexagonus. The molecular examinations showed the presence of A. phagocytophilum in 3.8%, B. microti in 24.5% and T. gondii in 4.5% of the all ticks. In addition, in the study area, there is a high potential risk of tick-borne infection by B. microti, and a low potential risk of exposure to A. phagocytophilum and T. gondii infection.

Molecular detection of tick-borne pathogens harbored by ticks collected from livestock in the Xinjiang Uygur Autonomous Region, China

Ticks carry and transmit a wide range of pathogens (bacteria, viruses, and protozoa) that are of importance to humans and animals globally. However, information about the tick-borne pathogens harbored by ticks in the Xinjiang Uygur Autonomous Region (XUAR), northwestern China, is scarce. This study investigated the occurrence of tick species of domestic animals and tick-borne pathogens by using morphological molecular identification and sequence analysis in Turpan, Qitai, Altay, Hejing, Nileke, and Zhaosu counties (XUAR). A total of 5822 adult ticks (females and males) from 12 tick species were identified from 5 animal species (cattle, goats, sheep, camels, and horses) in 6 counties in the XUAR. Collected tick species included Dermacentor marginatus (24.7 %), Dermacentor nuttalli (20.8 %), Hyalomma anatolicum (13.7 %), Dermacentor niveus (13.1 %), Haemaphysalis punctata (10.7 %), Dermacentor silvarum (7.1 %), Dermacentor pavlovskyi (3.9 %), Hyalomma asiaticum (2.2 %), Rhipicephalus pumilio (1.9 %), Rhipicephalus sanguineus sensu lato (0.7 %), Rhipicephalus turanicus (0.6 %), and Hyalomma asiaticum kozlovi (0.6 %). Furthermore, 750 partially engorged adult ticks (females and males), including H. anatolicum (250), D. nuttalli (250), and D. marginatus (250), were individually separated according to species and sampling site, used for DNA extraction, and then screened for tick-borne pathogens. The most common pathogen was Rickettsia raoultii (36.80 %), followed by Brucella sp. (26.2 %), Anaplasma ovis (22.4 %), Babesia caballi (14.8 %), Theileria equi (8.7 %), and Theileria ovis (8.5 %). The sequencing of 6 genes showed a 96–100 % nucleotide identity between the sequences in this study and those deposited in GenBank. This study provides a scientific reference for the prevention and control of tick-borne diseases in the XUAR.

Molecular detection of tick-borne pathogens in caracals (Caracal caracal) living in human-modified landscapes of South Africa

BackgroundWild carnivores living alongside humans and domestic animals are vulnerable to changes in the infectious disease dynamics in their populations. The aims of this study were to determine the prevalence and diversity of selected tick-borne pathogens (TBPs) of veterinary and/or zoonotic concern in wild populations of caracals (Caracal caracal) occurring in human-modified landscapes in South Africa. Using molecular techniques, we screened 57 caracal blood samples for infection by rickettsial bacteria and piroplasms in three regions of South Africa: rangeland in the Central Karoo (n = 27) and Namaqualand (n = 14) as well as the urban edge of the Cape Peninsula (n = 16) of South Africa. To characterise pathogen identity, we sequenced the 18S rRNA and 16S rRNA genes from positive samples and analysed sequences within a phylogenetic framework. We also examine the diversity of potential tick vectors.ResultsAll individuals tested were infected with at least one tick-borne pathogen. Pathogens included Hepatozoon felis, Babesia felis, Babesia leo and a potentially novel Babesia species. An Anaplasma species previously described in South African domestic dogs was also found in 88% of urban edge caracals. Higher rates of co-infection characterised urban edge caracals (81% vs 15% and 0% in the two rangeland populations), as well as a greater incidence of mixed infections. Host attached tick species include Haemaphysalis elliptica, an important pathogen vector among carnivore hosts.ConclusionsThis study confirms the occurrence of previously undocumented tick-borne pathogens infecting free-ranging caracals in human-modified landscapes. We identify clear differences in the pathogen profiles among our study populations and discuss the likely health costs to caracals living adjacent to urban areas.

Molecular detection of tick-borne pathogens in canine population and Rhipicephalus sanguineus (sensu lato) ticks from southern Metro Manila and Laguna, Philippines

BackgroundThe tropical climate of the Philippines and the high population of dogs, particularly in cities, favors the life-cycle of the brown dog tick, Rhipicephalus sanguineus (sensu lato), a vector of several canine tick-borne pathogens (TBPs) including zoonotic Rickettsia spp. Suspected cases of infections are commonly encountered in veterinary clinics, but the specific TBPs are rarely identified. Furthermore, infection with Rickettsia is not being clinically examined in dogs. In this study, the occurrence of TBPs in blood and ticks collected from household and impounded dogs in highly populated areas of the Philippines, Metro Manila, and the nearby province of Laguna, was examined.ResultsA total of 248 blood samples and 157 tick samples were subjected to PCR. First, samples were screened using primers for Anaplasma/Ehrlichia spp. and Babesia/Hepatozoon spp. Those that turned positive were further subjected to species-specific PCR. Rickettsia spp. were also detected through a nested PCR. Of the 248 blood samples, 56 (22.6%) were positive for Anaplasma/Ehrlichia spp., while 19 (7.6%) were positive for Babesia/Hepatozoon spp. Species-specific PCR revealed that 61 (23.4%) had a single TBP, with Ehrlichia canis being detected in 39 (15.7%) dogs, while 14 (5.6%) dogs were positive for different combinations of two to four TBPs. Rickettsia infection was detected in 6 (2.4%) dogs. In tick samples, 8 (3.2%) were positive for Ehrlichia/Anaplasma spp., while only 1 (0.63%) was positive for Babesia/Hepatozoon spp. As in the blood samples, E. canis was the most detected, being found in 5 (2%) samples. No tick samples tested positive for Rickettsia spp.ConclusionEhrlichia canis is the most common TBP affecting dogs in the Philippines. Co-infection with TBPs is quite common, hence testing for multiple TBPs is necessary. Through nested PCR, Rickettsia infection was detected in dogs, and to the authors’ knowledge, this study provides the first molecular evidence of Rickettsia infection in dogs in the Philippines.

Dog survey in Russian veterinary hospitals: tick identification and molecular detection of tick-borne pathogens

BackgroundSpecies of Canidae in Russia can be infested with up to 24 different tick species; however, the frequency of different tick species infesting domestic dogs across Russia is not known. In addition, tick-borne disease risks for domestic dogs in Russia are not well quantified. The goal of this study was to conduct a nationwide survey of ticks collected from infested dogs admitted to veterinary clinics in Russian cities and to identify pathogens found in these ticks.MethodsTicks feeding on dogs admitted to 32 veterinary clinics in 27 major cities across Russia were preserved in ethanol and submitted to a central facility for examination. After identification, each tick was evaluated for infection with known tick-borne pathogens using PCR.ResultsThere were 990 individual ticks collected from 636 dogs. All collected ticks belonged to the Ixodidae (hard ticks) and represented 11 species of four genera, Dermacentor, Ixodes, Rhipicephalus and Haemaphysalis. Four most common tick species were D. reticulatus, followed by I. persulcatus, I. ricinus and R. sanguineus. Ixodes persulcatus ticks were found to be infected with 10 different pathogens, and ticks of this species were more frequently infected than either D. reticulatus or I. ricinus. Ixodes persulcatus females were also more frequently co-infected with two or more pathogens than any other tick. Pathogenic species of five genera were detected in ticks: Anaplasma centrale, A. phagocytophilum and A. marginale; Babesia canis, B. microti, B. venatorum, B. divergens, B. crassa and B. vogeli; Borrelia miyamotoi, B. afzelii and B. garinii; Ehrlichia muris, E. canis and E. ruminantum; and Theileria cervi. Anaplasma marginale, E. canis, B. crassa, B. vogeli and T. cervi were detected in I. persulcatus, and Babesia canis in D. marginatum, for the first time in Russia.ConclusionsMultiple ticks from four genera and 11 species of the family Ixodidae were collected from domestic dogs across Russia. These ticks commonly carry pathogens and act as disease vectors. Ixodes persulcatus ticks present the greatest risk for transmission of multiple arthropod-borne pathogens.

Molecular detection of tick-borne pathogens in cattle from Southwestern Ethiopia.

Tick-borne diseases (TBDs) cause significant losses among livestock and impact the livelihoods of resource-poor farming communities worldwide. In Ethiopia, detailed studies on the epidemiology of tick-borne pathogens (TBPs) in cattle using sensitive molecular detection methods are scarce. The objective of this study was to determine the prevalence and species composition of bovine TBPs of veterinary significance in local cattle populations. A comprehensive cross-sectional epidemiological study was conducted in cattle populations of Illubabor zone in Southwestern Ethiopia from June to August 2013. For this purpose, blood samples were collected from 392 cattle. A combination of polymerase chain reaction (PCR) and a Reverse Line Blot (RLB) hybridization assay was employed for the detection of TBPs in these samples. The PCR/RLB results of the 392 blood samples indicated a high overall prevalence of 96.9% for TBPs, including Theileria mutans (66.1%), Theileria orientalis (51.8%), Anaplasma sp. Omatjenne (25.5%), Anaplasma marginale (14.5%), Babesia bigemina (14.0%) and Theileria velifera (13.0%) and minor occurrences of Ehrlichia ruminantium (0.5%) and Ehrlichia minasensis (0.26%). Moreover, three novel Anaplasma genotypes were detected in bovine blood samples. A phylogenetic analysis revealed that they most likely represent three, but at least two, new species. The prevalence of the three novel Anaplasma species, preliminary designated as Anaplasma sp. Hadesa, Anaplasma sp. Saso and Anaplasma sp. Dedessa, was 12.5%, 14.3% and 5.6%, respectively. Overall, a total of 227 cattle (57.9%) were found to be co-infected with two or more TBPs simultaneously and 86 different species combinations were observed. The findings show a very high burden of infection of cattle with TBPs in Ethiopia. The high frequency of co-infections suggests that clinical manifestations might be complex. Further research is required to determine the pathogenicity, host cell types and vector of the three novel Anaplasma species identified in this study.

Molecular detection of tick-borne pathogens in wild red foxes (Vulpes vulpes) from Central Italy

Spleen samples from 153 red foxes, shot during regular hunting season in the province of Pisa (Central Italy), were examined to detect DNA of Anaplasma phagocytophilum, Ehrlichia canis, Coxiella burnetii, Francisella tularensis, Hepatozoon canis and Babesia sp./Theileria sp.DNA of vector-borne pathogens was detected in 120 (78.43%; 95% CI: 71.06–84.66%) foxes. Specifically, 75 (49%; 95% CI: 40.86–57.22%) animals scored PCR-positive per H. canis, 68 (44.44%; 95% CI: 36.42–52.69%) for E. canis, 35 (22.88%; 95% CI: 16.48–30.35%) for piroplasms (Theileria annae), 3 (1.96%; 95% CI: 0.41–5.62%) for C. burnetii and 1 (0.65%; 95% CI: 0.02–3.59%) for A. phagocytophilum. No positive reaction was observed for F. tularensis. Fifty-six animals (36.6%; 95% CI: 28.97–44.76%) were positive for two or three pathogens. Red foxes result to be involved in the cycle of vector-borne pathogens that are associated to disease in dogs and humans.

First molecular detection of tick-borne pathogens in dogs from Jiangxi, China

In this study, blood samples obtained from 162 dogs in Jiangxi, China, were employed in molecular screening of canine tick-borne pathogens by PCR and sequencing. Babesia spp. gene fragment was detected in 12 (7.41%) dogs. All samples were negative for Hepatozoon spp., Ehrlichia canis, Coxiella spp., Borrelia spp., Rickettsia spp. and Anaplasma platys. Species-specific PCR analysis further confirmed that 8 (4.94%) and 4 (2.47%) dogs were infected by Babesia canis vogeli and Babesia gibsoni, respectively. Based on our analyses, Babesia spp. infection in Jiangxi appeared not related to age, gender, breed, usage, activity and health status or tick infestation history of the dogs. This is the first molecular report of Babesia canis vogeli and Babesia gibsoni in dogs from Jiangxi, China.