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Borrelia persica infection in wild carnivores in Israel: molecular characterization and new potential reservoirs

Parasites & Vectors volume 16, Article number: 337 (2023) Cite this article

Abstract

Background

Borrelia persica causes tick-borne relapsing fever in Israel, the eastern Mediterranean basin, and Asia. Relapsing fever is associated with severe illness and potentially death in humans and animals. Since B. persica infection has rarely been described in wild animals, the aim of this study was to evaluate the prevalence of infection with B. persica in wild carnivores in Israel.

Methods

Spleen and blood clot samples from wild carnivores, which underwent necropsy, were tested for the presence of Borrelia DNA by real-time polymerase chain reaction (PCR). PCR products were sequenced, and the spirochete loads were quantified using a specific quantitative PCR (qPCR).

Results

A total of 140 samples from 74 wild carnivores were analyzed for the presence of Borrelia DNA. Six out of the 74 (8.1%) animals were found positive for B. persica by PCR and sequencing of the flagellin B gene, of which 4/74 (5.4%) were also positive by PCR for the glycerophosphodiester phosphodiesterase (glpQ) gene. Positive samples were obtained from three European badgers, and one striped hyena, golden jackal, and red fox each. All B. persica-positive animals were young males (P < 0.0001). Quantifiable results were obtained from 3/5 spleen and 4/5 blood samples. The spirochete loads in the blood were significantly higher than those found in the spleen (P = 0.034).

Conclusions

The prevalence of B. persica infection found in wild carnivores brought for necropsy was unexpectedly high, suggesting that this infection is widespread in some wild animal species in Israel. This is the first report of B. persica infection in the European badger and striped hyena. These carnivores have a wide geographical range of activity, and the results of this survey raise the possibility that they may serve as reservoir hosts for B. persica.

Graphical Abstract

Background

Relapsing fever is a zoonotic infectious disease caused by spirochetes from the genus Borrelia. The disease in humans is characterized by recurrent fever episodes, headaches, lethargy, tachycardia, conjunctivitis, hepatomegaly, splenomegaly, pigmenturia, vomiting, myalgia, and arthralgia [1, 2]. Relapsing fever can be divided into the louse-borne relapsing fever, caused by Borrelia recurrentis and transmitted by the human body louse Pediculus humanus, and tick-borne relapsing fever (TBRF), which is mostly transmitted by argasid ticks from the genus Ornithodoros [3, 4]. The TBRF agents include also a small group of species, which are transmitted by ixodid ticks, such as Borrelia miyamotoi transmitted by ticks of the genus Ixodes, Borrelia theileri transmitted by Rhipicephalus spp., and Borrelia lonestari transmitted by Amblyomma americanum [5].

Relapsing fever caused by Borrelia persica has been reported in humans from Iran, Pakistan, several Central Asian countries, and Egypt, and it is the only reported endemic relapsing fever species in Israel [6,7,8,9]. Mortality due to B. persica infection in humans is low in patients treated with antibiotics and monitored during therapy; however, fatal infections with severe complications have been reported in Israel [10, 11]. Ornithodoros tholozani, the vector of B. persica, is common in Israel and is among the few soft tick species found in the country [12]. The geographical distribution of O. tholozani extends from India and Central Asia to Egypt and overlaps the distribution of B. persica [13, 14]. Like other Ornithodoros spp., O. tholozani ticks are rapid blood feeders, with all life stages feeding on warm-blooded mammals, including humans, rodents, cattle, small ruminants, camels, foxes, jackals, porcupines, and hedgehogs [4, 13]. Ornithodoros tholozani is commonly found in shaded locations, including caves, ruins, and rock crevices [14]. The reported incidence of TBRF in civilians in Israel has declined from 0.35 cases per 100,000 inhabitants in the years 1975 to 1985, to 0.11/100,000 from 1986 to 2003. Nevertheless, the incidence among Israeli military personnel remained stably high with an annual average of 6.4 cases per 100,000 [15].

Rodents are considered the main reservoir hosts for the North American TBRF species Borrelia hermsii, Borrelia parkeri, and Borrelia turicatae, as well as for the Old World species Borrelia hispanica and Borrelia crocidurae [16,17,18,19]. Infection with B. persica in animals in Israel has been described in domestic dogs and cats, in which the infection is associated with clinical disease and clinicopathological findings such as fever, lethargy, anorexia, anemia, and thrombocytopenia, and can be fatal [20, 21]. Clinical manifestations of relapsing fever were also observed in dogs infected with the North American species B. turicatae and B. hermsii, and in cats infected with B. hispanica [22,23,24]. Previously, B. persica was identified by molecular methods in several wildlife species in Israel, mostly herbivores and insectivores [25, 26].

According to the World Health Organization (WHO) reports, zoonotic diseases are responsible for about 60% of all emerging infectious diseases (EID), with an estimated global morbidity of 107 patients and several million human fatalities annually [27]. Wild animals were found to play a major role in the global burden caused by EID, with more than 70% of zoonotic EID originating in wildlife [28]. Wild carnivores are an important part of the sylvatic cycle in different zoonotic diseases, such as rabies, cystic echinococcosis, and trichinellosis [29,30,31]. The close proximity between wild animals and humans, even in urban areas, may contribute to the role of the former in human infection [32]. Therefore, knowledge of the prevalence of B. persica infection in wild carnivores in Israel and their possible part in the epidemiology of relapsing fever is essential. The aims of this study were to survey a variety of wild carnivore species in Israel for infection with B. persica and to evaluate the bacterial loads in infected animals. Sampling of Indian crested porcupines (Hystrix indica), which are large omnivore rodents that prey on small vertebrates [33], was also included in the study, because they were identified in a previous study as the main source for blood meal in O. tholozani ticks surveyed in Israel [25].

Methods

Sample collection and DNA extraction

Spleen and blood samples were collected from wild carnivores that underwent post-mortem examination at the Pathology Department of the Kimron Veterinary Institute in Beit Dagan, Israel. Data collected included the carcass's geographical point of collection, species, sex, presumptive age, and necropsy report. There were no data regarding the animal’s clinical signs prior to death. Samples were preserved at −80 °C until DNA extraction was conducted. DNA was extracted from 10 mg of spleen and 100 µl blood using a commercial kit (DNeasy Blood & Tissue Kit, Qiagen, Germany) following the manufacturer's protocol, with some modifications; spleen samples were washed with phosphate-buffered saline (PBS) and mechanically crushed before the standard protocol was performed.

Molecular identification and genetic characterization of Borrelia spp.

Screening of samples for Borrelia DNA was performed by real-time quantitative polymerase chain reaction (qPCR) using primers Bfpbu (5′-GCTGAAGAGCTTGGAATGCAACC-3′) and Bfpcr (5′-TGATCAGTTATCATTCTAATAGCA-3′) targeting a 346-base-pair (bp) fragment of the flagellin (flaB) gene of Borrelia [34]. Positive samples were also tested for the relapsing fever-specific gene glycerophosphodiester phosphodiesterase (glpQ) by real-time PCR using primers 510f (5′-AAAACCCTTTTGGCATAAACAACA-3′) and 770R (5′-CCAGGGTCCAATTCCGTCAG-3′) targeting a 280-bp fragment of the gene [35]. The flaB and glpQ real-time PCR protocols were carried out with an initial hold for 3 min at 95 °C, followed by 45 cycles of 15 s at 95 °C, 30 s at 58 °C, and 10 s at 72 °C. The melting phase started at 60 °C, each step rising by 0.3 °C, and finished at 95 °C with a hold for 90 s at the first step and 5 s at the subsequent steps. Each reaction was performed in 20 μl reaction volume containing 4 μl of DNA, 0.5 μM of each primer, 0.6 μl of syto9 (Invitrogen, CA, USA.), 4.4 μl of distilled extra-pure water, and 10 μl of DreamTaq PCR Master Mix (Thermo Scientific, Loughborough, UK). DNA extracted from B. persica culture grown in the authors' laboratory was used as a positive control. All reactions were performed in the Life Technologies StepOnePlus Real-Time PCR System (Thermo Fisher Scientific, Waltham, MA, USA). Samples were considered positive when the melting temperature (Tm) was equal to the positive control Tm and found to be 82 °C for the flaB gene and 77.6 °C for the glpQ gene. Positive PCR products were sequenced from both directions using the respective forward and reverse primers. No purification was made prior to sequencing. Sequencing was performed by the BigDye Terminator cycle sequencing chemistry from Applied Biosystems ABI 3700 DNA Analyzer (ABI, Carlsbad, CA, USA) and the ABI Data collection and Sequence Analysis software at the Center for Genomic Technologies, Hebrew University of Jerusalem, Israel. DNA sequences were aligned using the Molecular Evolutionary Genetics Analysis (MEGA) software, version 11.013 [36]. Sequences were further compared with the GenBank database using the BLAST algorithm (National Center for Biotechnology Information, Bethesda, MD, USA; http://blast.ncbi.nlm.nih.gov/Blast.cgi). Species-level identification was obtained according to the closest BLAST match when sequences showed identity higher than 99% to GenBank sequences.

Quantification of B. persica in spleen and blood samples

Quantitative PCR was performed on the positive spleen and blood samples by amplifying a 346-bp fragment of the flaB gene of Borrelia [34]. The purpose of quantifying the B. persica bacterial load was to validate infection by characterizing the numbers of spirochetes in the blood and spleen, and to compare the bacterial load in the different tissues.

Standard curves for spleen and blood were generated using 10-fold dilutions of B. persica grown in culture, from an isolate originally cultured from a cat in Israel and designated as the B. persica LMU-C01 strain [37]. Cultured spirochetes were counted by dark-field microscopy using a Neubauer chamber cell (Neubauer improved, Boeco, Germany). Ten milligrams of spleen and 100 µl of blood from a golden jackal which tested negative for the presence of Borrelia spp. by PCR were spiked with 6 × 107 cultured B. persica organisms. DNA was extracted from the spiked spleen and blood using the DNeasy Blood & Tissue Kit (Qiagen, Germany). The extracted DNA was diluted with ultra-pure DNA-free water to reach the concentrations of 6 × 107 to 6 × 101 spirochetes per sample. Standard curves were obtained by running duplicates of the spiked spleen and blood with culture dilution reference points and plotting their cycle threshold (Ct) values against the log of B. persica absolute numbers using the StepOne software version 2.2.2 (Applied Biosystems, Thermo Fisher Scientific, Foster City, CA, USA). Real-time PCR and reaction mix protocols were carried out as described for the flaB gene.

In order to correct B. persica quantities to host DNA quantity in the spleen and blood samples, a real-time PCR was performed to detect mammalian DNA by targeting a 150-bp fragment of vertebrate 18S rRNA gene using universal primers 0033F (5′-TTCTAGAGCTAATACATGCCGA-3′) and 0049R (5′-CGAGGTTATCTAGAGTCACC-3′) specific for vertebrates as previously described [38]. Real-time PCR, the melting phase, and the components of the reaction were the same as for the amplification of the flaB and glpQ genes. DNA extracted from a culture of B. persica was used as a negative control. To obtain the actual loads of spirochetes, B. persica flaB Ct values were adjusted by calculating the difference between the actual mammal 18S rRNA gene Ct obtained for each sample and the average mammal 18S rRNA gene Ct values, and adding this value to the Borrelia flaB Ct value for each sample. Finally, the B. persica load (absolute number per millimeter of blood and milligram of spleen) in each sample was calculated according to the linear regression equation of the standard curve (Cт = m [log (Qty)] + b; where m is the slope, b is the y-intercept, and Qty is the standard quantity), which was obtained using StepOne version 2.2.2 software (Thermo Fisher Scientific).

Statistical analysis

Statistical analysis was performed using the SPSS software version 26.0 (SPSS IBM, Armonk, NY, USA). Comparisons of sex and age differences between positive and negative animals were tested using the Chi-square test. The continuity correction was employed when 2 × 2 tables were used. Bacterial loads in blood samples as compared to spleen samples were tested by the non-parametric Mann–Whitney test for two independent variables. A Pearson correlation coefficient was computed to assess the linear relationship between spirochetal loads in the spleen and in the blood. Statistical significance was defined as P < 0.05. Maps showing the collection sites were constructed using the ArcGIS Map 10.0 software (Esri, Redlands, CA, USA).

Results

A total of 140 samples (68 blood and 72 spleen samples) from 74 wild carnivores, obtained by the Pathology Department at the Kimron Veterinary Institute from November 2020 to September 2022, were analyzed for the presence of B. persica DNA. The specimens originated from wild animals of five families and eight species, including the golden jackal (Canis aureus), red fox (Vulpes vulpes), wolf (Canis lupus), Egyptian mongoose (Herpestes ichneumon), striped hyena (Hyaena hyaena), Indian crested porcupine (Hystrix indica), European badger (Meles meles), and beech marten (Martes foina) (Table 1). The most frequent cause of admission for post-mortem examination was culling wild carnivores as part of a rabies monitoring programs (79.7%), followed by road kills (19.0%) and predation (1.3%).

Table 1 Wild animal species sampled and rate of infection with Borrelia persica in blood and spleen samples
Full size table

Out of 74 wild animals, six (8.1%) were found positive for the presence of Borrelia spp. DNA by PCR amplification and sequencing of the flaB gene. All positive samples showed > 99% identity to B. persica GenBank accession number KY964442.1 previously amplified from a rock hyrax in Israel. All samples were also tested by the glpQ PCR, and four (5.4%) were found positive and showed > 99% identity to B. persica GenBank accession number MF401448, which was also amplified from a rock hyrax in Israel. All four animals positive for the glpQ PCR were also positive for the flaB PCR. All PCR-positive animals had both spleen and blood samples available. Four out of six (67%) positive animals were positive in both spleen and blood, 1/6 (16.7%) was positive only in the spleen, and 1/6 (16.7%) was positive only in the blood.

Positive samples (Table 1) were obtained from three European badgers (M. meles), one striped hyena (H. hyaena), one golden jackal (C. aureus), and one red fox (V. vulpes). Overall, one of three (33.3%) striped hyenas was found positive, followed by the European badgers (3/16, 18.75%), red fox (1/8, 12.5%), and golden jackal (1/40, 2.5%).

No significant differences were found in the prevalence between the positive animals (χ2 = 6.237, df = 3, P = 0.101).

Sex and age were known for 54 and 38 animals, respectively. Twenty-eight out of 54 (51.9%) animals for which the sex was documented were females, and 26/54 (48.1%) were males. Furthermore, 29/38 (76.3%) animals, for which the age was documented, were adults, whereas 9/38 (23.7%) were juvenile. All positive animals were young-age males (Table 2). Sex and age proportions in the B. persica-positive animals were found significantly different from those proportions in the general study population (χ2 = 5.120, df = 1. P = 0.024; χ2 = 18.218, df = 1, P < 0.0001, respectively). Eleven out of 74 animals tested were found infested with ixodid ticks.

Table 2 Sex and age details available in the study population and the Borrelia persica-positive animals
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The geographical distribution of the animals tested ranged from Metula and Ein Zivan in northern Israel to Mount Sagi in the south of the country (Fig. 1). Positive animals were found in six out of 51 distinct geographical locations, including three animals from northern Israel and three from the central part of the country (Fig. 1, Table 1).

Fig. 1
figure 1

Geographical distribution of sampled wild carnivores indicating the presence or absence of infection with B. persica. The locations of positive animals are marked in red dots, and locations where negative animals were found are marked in black dots. Positive animals are highlighted with its species name

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Quantifiable numbers of B. persica were available from 3/5 (60%) PCR-positive spleen samples and 4/5 (80%) blood samples (Table 3). The minimal detection limit was determined by the concentration for which the assay maintained linearity for spleen and for blood samples and was found to be as low as 63 spirochetes per 1 mg of spleen and 6333 spirochetes per 1 ml of blood. The minimal detection limit was equal for the spleen and the blood as the concentrations for these two tissues were per 10 mg for the spleen and per 1 ml for the blood tissue. After normalization with mammalian DNA, the median B. persica load in the three quantifiable spleen samples was 2903 spirochetes/mg spleen (range 244–12,673 spirochetes/mg), and for the four quantifiable blood samples, it was 257,086 spirochetes/ml blood (range 14,514–1,547,764 spirochetes/ml). Two spleen and one blood samples harbored a small quantity of B. persica DNA that was below the quantifiable detection limit of the assay (Table 3). The absolute numbers of B. persica spirochetes based on the flaB amplification were significantly higher in the blood compared to the spleen (U(7) < 0.0001, Z = −2.121, P = 0.034). In addition, an almost significant correlation was found between spleen and blood spirochete loads of the same animals (R2 = 0.997, P = 0.051).

Table 3 Borrelia persica loads in blood and spleen samples
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Phylogenetic analysis based on a 229-bp segment of the flaB gene sequence (Fig. 2) revealed that all B. persica sequences from this study clustered together. Four sequences of the B. persica-positive animals clustered together with a B. persica genotype II sequence, previously amplified from a human patient in Israel (GenBank DQ679907.1), whereas two sequences from positive animals clustered together with a B. persica genotype I sequence from an O. tholozani tick collected in Israel (GenBank DQ679905.1) [39]. None of the sequences from the wildlife study clustered with the B. persica genotype III sequence, which originated from a human patient in Israel (GenBank DQ673617). All B. persica sequences clustered separately from other Old World relapsing fever Borrelia spp. including B. recurrentis, B. duttonii, and B. crocidurae, except for Borrelia caucasica, which clustered together with B. persica genotype III sequence. North American relapsing fever species including B. parkeri and B. turicatae also clustered separately. TBRF species which are transmitted by ixodid hard ticks including B. lonestari and B. theileri clustered together, but separately from B. miyamotoi, which clustered together with B. hermsii.

Fig. 2
figure 2

A maximum likelihood phylogram comparing 229-bp DNA fragment sequences of the flaB gene from the wild carnivores included in the study to sequences from other B. persica and other Borrelia spp. GenBank accessions. New sequences derived from this study are marked with black diamond squares. Note the division into B. persica genotypes marked in Roman numerals. The GenBank accession numbers, species of infected host, and country of origin are included for each sequence. The Tamura 3-parameter model was used in the construction of this phylogram with bootstrap performed on 1000 replicates, and values higher than 60% are indicated

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Phylogenetic analysis based on a 260-bp segment of the glpQ gene (Fig. 3) also revealed that sequences from B. persica-positive wild animals clustered with each other and with other B. persica sequences from a human, cat, and an O. tholozani tick. As in the flaB phylogram, B. persica glpQ sequences clustered separately from Old World relapsing fever Borrelia spp. including B. hispanica, B. duttonii, B. crocidurae, and B. recurrentis. The Old World species including B. persica also clustered separately from American relapsing fever Borrelia spp., namely B. hermsii, B. parkeri, and B. turicatae.

Fig. 3
figure 3

A maximum likelihood phylogram comparing 260-bp DNA fragment sequences of the glpQ gene from the wild carnivores included in the study to sequences from other B. persica and other Borrelia spp. GenBank accessions. New sequences derived from this study are marked with black diamond squares. The GenBank accession numbers, species of infected host, and country of origin are included for each sequence. The Tamura-3-Parameter model was used in the construction of this phylogram with bootstrap performed on 1000 replicates, and values higher than 70% are indicated

Full size image

Discussion

This study describes infection with the TBRF spirochete B. persica in 8% of wild carnivores examined in Israel, and includes the first report of this infection in the European badger and striped hyena. Infection with B. persica accompanied by clinical disease was previously described in humans, dogs, and cats [15, 20]. A previous study from Israel, in which samples from domestic dogs and cats admitted to a veterinary teaching hospital for treatment unrelated to relapsing fever were tested using a PCR-based protocol, found that the prevalence of B. persica was 1.9% in the dog group and 2.9% in the cat group [21]. Other studies pertaining to animals from Israel reported that B. persica infection was detected by PCR in 7% of the screened wild animals, including the social vole (Microtus socialis), fat sand rat (Psammomys obesus), Cairo spiny mouse (Acomys cahirinus), red fox (V. vulpes), and golden jackal (C. aureus), and in 8% of tested rock hyraxes (Procavia capensis) in a second study [25, 26]. Clinical signs of disease have not been described in these previous reports of infection in wildlife in Israel. The infected animals in the current study belong to three different carnivoran families and include the golden jackal, red fox, and the newly reported European badger and striped hyena.

Wild animals are considered reservoirs for several viral, bacterial, protozoal, and helminthic zoonoses [40, 41]. Over the past decade, 51.1% of the reported cases of rabies in Israel were diagnosed in wild carnivores, including golden jackals, red foxes, wolves, and European badgers [42]. The European badger is widely distributed across Europe, the Middle East, and several countries in Western Asia, and it is known to serve as a reservoir host for zoonotic pathogens, such as the tuberculosis bacterium Mycobacterium bovis, the Lyme borreliosis spirochete Borrelia afzelii, and species of the nematode Trichinella [41, 43,44,45]. A previous study from Israel demonstrated that 72 out of 2777 (2.6%) O. tholozani ticks collected in 16 caves were found positive for B. persica DNA by PCR. Out of 72 infected ticks, 50 were positive for the presence of host blood. Two ticks (4%) were found positive for B. persica infection with host blood from the European badger [25]. To the best of our knowledge, our findings represent the first published data regarding infection of the European badger with any relapsing fever Borrelia spp. These findings in which three of 16 badgers were naturally infected with B. persica and the fact that O. tholozani ticks with badger blood were found to be infected with B. persica in a previous study strengthen the suggestion that the European badger may serve as a natural reservoir host for B. persica transmission.

The striped hyena is a solitary predator distributed over Northern and Eastern Africa, the Middle East, and India [46]. Its individual range of activity is very wide and documented to reach 72 and 44 km2 for males and females, respectively [47]. In Israel, an increase in hyena population density has been observed over the past few decades, with about two individuals per 100 km2 compared to 0.06 individuals per 100 km2 in the 1970s [48]. In contrast to the European badger, hyenas are less common reservoirs for human infections, with only a few reports of zoonotic pathogen infection in wild hyenas [49, 50]. This is the first report of infection with Borrelia spp. in the striped hyena. In a previous study, three O. tholozani nymphs were found positive for the presence of host blood from striped hyenas by PCR [25]. These findings support the possibility that the striped hyena may serve as a reservoir host for B. persica infection as well. The striped hyena is also capable of distributing B. persica between distant locations due to its wide range of activity.

The European badger and the striped hyena both have a long lifespan of about 8–15 years [51, 52], and when their activity ranges and geographical distributions are taken into account, it is suggested that these newly found hosts for B. persica could serve as potential reservoirs for transmission of the pathogen to humans and animals via ticks.

The geographical distribution of samples collected during this study covers large areas of Israel and overlaps much of the distribution area of O. tholozani, which is found throughout Israel except for the southern part of the Negev desert in southern Israel [14]. This is supported by a previous study on the prevalence of TBRF in Israel which demonstrated a relationship between this geographical area and the presence of clinical disease cases in humans [53]. Borrelia persica-positive wild carnivores were also distributed widely across Israel in the current study, with representation for the Jerusalem, Tel Aviv, the Western Galilee, and the Golan Heights areas in central and northern Israel.

Age and sex were found significant factors for infection with B. persica in our study, with only young males found infected with B. persica [20, 26]. In a previous study that compared the prevalence of TBRF in children and adult humans, the average age among TBRF patients was 21.3 years old with 33% of the patients under 18 years old. Furthermore, out of 92 patients, 82% were males [54]. A likely explanation for young animals being more susceptible to B. persica infection is the possible development of resistance in older animals after initial infection at a young age, incomplete maturity of the immune system, or immune suppression due to co-infection, with reduced capability of successful defense against this infection. Infectious agents, such as the canine distemper virus, are associated with immunodeficiency in young animals and were described in the European badger, red fox, and golden jackal [55,56,57].

Both spleen and blood samples were found suitable for the molecular detection of B. persica infection. This is similar to findings from a previous study in hyraxes where the spirochete burden in the blood was up to 1000 times that in the spleen [26]. In the previous study on the prevalence of B. persica in rock hyraxes, spirochete loads were quantified by the measurement of the 16S rRNA gene copies and ranged from 2.3 × 104 to 1 × 106 copies in the spleen and from 5 × 106 to 9.2 × 108 in the blood [26]. Nevertheless, this is the first study to report an absolute quantification of B. persica in tissues using an in vitro cultivated culture. Absolute B. persica quantities in the spleen and blood were measured using calibration curves of spiked spleen and blood with a known quantity of spirochetes. The spiking method was used in order to balance the effect of internal inhibitors in the different tissues. The spiking process was done using the same culture and DNA extraction method, and therefore the results accurately represent the absolute quantity of B. persica spirochetes in the infected tissues.

Wild carnivore populations are expanding rapidly in many parts of Israel and encroaching into human habitation [48, 58]. The potential role of wild carnivores as reservoirs of B. persica and their proximity to human settlements may be considered as a risk factor for human and companion animal infection, which control programs for TBRF should address. In contrast to the limited data regarding TBRF in wild carnivores, the occurrence of Lyme borreliosis in those animals was more comprehensively studied, and evidence of infection with Borrelia burgdorferi, B. afzelii, and Borrelia garinii was demonstrated in several wild carnivore species, including the raccoon (Procyon lotor), red fox, raccoon dog (Nyctereutes procyonoides), European badger, pine marten (Martes martes), and stone marten (M. foina) [59]. Control and management programs aim to reduce infection rates of various infectious agents, including Lyme borreliosis, in wild carnivores through oral vaccination programs. In the case of Lyme borreliosis, studies in experimentally infected laboratory mice have shown protection via oral-route vaccination with live recombinant Escherichia coli or vaccinia virus expressing the OspA protein of B. burgdorferi [60]. This strategy may also be beneficial for protection against B. persica infection in wild carnivores.

The present study had several limitations, which include a relatively limited number of wild carnivores surveyed, a small representation of individual animal species, and missing information on some of the animals. Despite these limitations, the study revealed important information on the high prevalence of B. persica infection in wild carnivores in Israel and demonstrated that this infection is widely spread in the surveyed areas.

Availability of data and materials

All data generated or analyzed during this study are included in this published article.

References

  1. Dworkin MS, Schwan TG, Anderson DE, Borchardt SM. Tick-borne relapsing fever. Infect Dis Clin North Am. 2008;22:449–68.

    PubMed  PubMed Central  Google Scholar 

  2. Barbour AG, Hayes SF. Biology of Borrelia species. Microbiol Rev. 1986;50:381–400.

    CAS  PubMed  PubMed Central  Google Scholar 

  3. Parola P, Raoult D. Ticks and tickborne bacterial diseases in humans: an emerging infectious threat. Clin Infect Dis. 2001;32:897–928.

    CAS  PubMed  Google Scholar 

  4. Hoogstraal H. Argasid and nuttalliellid ticks as parasites and vectors. Adv Parasitol. 1985;24:135–238.

    CAS  PubMed  Google Scholar 

  5. Talagrand-Reboul E, Boyer PH, Bergström S, Vial L, Boulanger N. Relapsing fevers: Neglected tick-borne diseases. Front Cell Infect Microbiol. 2018;8:98.

    PubMed  PubMed Central  Google Scholar 

  6. Masoumi Asl H, Goya MM, Vatandoost H, Zahraei SM, Mafi M, Asmar M, et al. The epidemiology of tick-borne relapsing fever in Iran during 1997–2006. Travel Med Infect Dis. 2009;7:160–4.

    CAS  PubMed  Google Scholar 

  7. Davis GE, Hoogstraal H. Biology of the spirochete Borrelia persica, found in the tick Ornithodorus tholazani (Argasinae) collected in the governorat of the Western Egyptian Desert; comments on the distribution and ecology of the vector tick. Ann Parasitol Hum Comp. 1956;31:147–54.

    CAS  PubMed  Google Scholar 

  8. Muigg V, Seth-Smith HMB, Goldenberger D, Egli A, Nickel B, Dürig R, et al. Tick-borne relapsing fever caused by Borrelia persica in traveler to central Asia, 2019. Emerg Infect Dis. 2020;26:424–6.

    Google Scholar 

  9. Safdie G, Farrah IY, Yahia R, Marva E, Wilamowski A, Sawalha SS, et al. Molecular characterization of Borrelia persica, the agent of tick borne relapsing fever in Israel and the Palestinian authority. PLoS ONE. 2010;5:e14105.

    PubMed  PubMed Central  Google Scholar 

  10. Yossepowitch O, Gottesman T, Schwartz-Harari O, Soroksky A, Dan M. Aseptic meningitis and adult respiratory distress syndrome caused by Borrelia persica. Infection. 2012;40:695–7.

    CAS  PubMed  Google Scholar 

  11. Yagupsky P, Moses S. Neonatal Borrelia species infection (relapsing fever). Am J Dis Child. 1985;139:74–6.

    CAS  PubMed  Google Scholar 

  12. Filippova NA, Wilamowski A, Bromley-Schnur H, Uspensky I. Ticks of the subgenus Argas and findings of Argas latus in Israel. Med Vet Entomol. 1999;13:212–3.

    CAS  PubMed  Google Scholar 

  13. Assous MV, Wilamowski A. Relapsing fever borreliosis in Eurasia - Forgotten, but certainly not gone! Clin Microbiol Infect. 2009;15:407–14.

    CAS  PubMed  Google Scholar 

  14. Avivi A, Warburg M, Galun R. Ecological studies on the cave tick Ornithodoros tholozani and its distribution in Israel. Isr J Entomol. 1973;8:109–29.

    Google Scholar 

  15. Sidi G, Davidovitch N, Balicer RD, Anis E, Grotto I, Schwartz E. Relapsing fever in Israel. Emerg Infect Dis. 2005;11:2003–5.

    Google Scholar 

  16. Nieto NC, Teglas MB. Relapsing fever group Borrelia in southern California rodents. J Med Entomol. 2014;51:1029–34.

    PubMed  Google Scholar 

  17. Burgdorfer W, Mavros AJ. Susceptibility of various species of rodents to the relapsing fever spirochete Borrelia hermsii. Infect Immun. 1970;2:256–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  18. Trape JF, Diatta G, Arnathau C, Bitam I, Sarih M, Belghyti D, et al. The epidemiology and geographic distribution of relapsing fever borreliosis in West and North Africa, with a review of the Ornithodoros erraticus complex (Acari: Ixodida). PLoS ONE. 2013;8:e78473.

    CAS  PubMed  PubMed Central  Google Scholar 

  19. Diatta G, Duplantier JM, Granjon L, Bâ K, Chauvancy G, Ndiaye M, et al. Borrelia infection in small mammals in West Africa and its relationship with tick occurrence inside burrows. Acta Trop. 2015;152:131–40.

    PubMed  Google Scholar 

  20. Baneth G, Nachum-Biala Y, Halperin T, Hershko Y, Kleinerman G, Anug Y, et al. Borrelia persica infection in dogs and cats: Clinical manifestations, clinicopathological findings and genetic characterization. Parasit Vectors. 2016;9:244.

    PubMed  PubMed Central  Google Scholar 

  21. Baneth G, Dvorkin A, Ben-Shitrit B, Kleinerman G, Salant H, Straubinger RK, et al. Infection and seroprevalence of Borrelia persica in domestic cats and dogs in Israel. Parasit Vectors. 2022;15:102.

    CAS  PubMed  PubMed Central  Google Scholar 

  22. Whitney MS, Schwan TG, Sultemeier KB, McDonald PS, Brillhart MN. Spirochetemia caused by Borrelia turicatae infection in 3 dogs in Texas. Vet Clin Pathol. 2007;36:212–6.

    PubMed  Google Scholar 

  23. Kelly AL, Raffel SJ, Fischer RJ, Bellinghausen M, Stevenson C, Schwan TG. First isolation of the relapsing fever spirochete, Borrelia hermsii, from a domestic dog. Ticks Tick Borne Dis. 2014;5:95–9.

    PubMed  Google Scholar 

  24. Breitschwerdt EB, Nicholson WL, Kiehl AR, Steers C, Meuten DJ, Levine JF. Natural infections with Borrelia spirochetes in two dogs from Florida. J Clin Microbiol. 1994;32:352–7.

    CAS  PubMed  PubMed Central  Google Scholar 

  25. Kleinerman G, Eshed T, Nachum-Biala Y, King R, Baneth G. Transmission of the human relapsing fever spirochete Borrelia persica by the Argasid Tick Ornithodoros tholozani involves blood meals from wildlife animal reservoirs and mainly transstadial transfer. Appl Environ Microbiol. 2021;87:1–14.

    Google Scholar 

  26. Kleinerman G, King R, Nachum-Biala Y, Baneth G. Borrelia persica infection in rock hyraxes. Ticks Tick Borne Dis. 2018;9:382–8.

    PubMed  Google Scholar 

  27. Dong X, Soong L. Emerging and re-emerging zoonoses are major and global challenges for public health. Zoonoses. 2021;1:1–2.

    Google Scholar 

  28. Jones KE, Patel NG, Levy MA, Storeygard A, Balk D, Gittleman JL, et al. Global trends in emerging infectious diseases. Nature. 2008;451:990–3.

    CAS  PubMed  PubMed Central  Google Scholar 

  29. Mandal S, Deb MM. Human cystic echinococcosis: Epidemiologic, zoonotic, clinical, diagnostic and therapeutic aspects. Asian Pac J Trop Med. 2012;5:253–60.

    PubMed  Google Scholar 

  30. Lahmar S, Torgerson PR, Mhemmed H, Tizaoui L, Mhadhbi N, Bani A, et al. Cystic echinococcosis and other helminth infections of wild boar in northeastern and northwestern regions of Tunisia. Parasitology. 2019;146:1263–74.

    CAS  PubMed  Google Scholar 

  31. David D, Dveres N, Yakobson BA, Davidson I. Emergence of dog rabies in the Northern region of Israel. Epidemiol Infect. 2009;137:544–8.

    CAS  PubMed  Google Scholar 

  32. Bombieri G, Delgado M, Russo LF, Garrote PJ, López-bao JV, Fedriani JM, et al. Patterns of wild carnivore attacks on humans in urban areas. Sci Rep. 2018;8:17728.

    CAS  PubMed  PubMed Central  Google Scholar 

  33. Kadhim AHH. Distribution and reproduction of the Indian crested porcupine Hystrix indica (hystricidae: Rodentia) in Iraq. Zool Middle East. 1997;15:9–12.

    Google Scholar 

  34. Fukunaga M, Ushijima Y, Aoki Y, Talbert A. Detection of Borrelia duttonii, a tick-borne relapsing fever agent in central Tanzania, within ticks by Flagellin gene-based nested polymerase chain reaction. Vector-Borne Zoonotic Dis. 2001;1:331–8.

    CAS  PubMed  Google Scholar 

  35. Potkonjak A, Kleinerman G, Gutiérrez R, Savić S, Vračar V, Nachum-Biala Y, et al. Occurrence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks with first identification of Borrelia miyamotoi in Vojvodina. Serbia Vector-Borne Zoonotic Dis. 2016;16:631–5.

    PubMed  Google Scholar 

  36. Tamura K, Stecher G, Peterson D, Filipski A, Kumar S. MEGA6: Molecular evolutionary genetics analysis version 6.0. Mol Biol Evol. 2013;30:2725–9.

    CAS  PubMed  PubMed Central  Google Scholar 

  37. Schwarzer S, Margos G, Overzier E, Fingerle V, Baneth G, Straubinger RK. Borrelia persica: In vitro cultivation and characterization via conventional PCR and multilocus sequence analysis of two strains isolated from a cat and ticks from Israel. Ticks Tick Borne Dis. 2015;6:751–7.

    PubMed  Google Scholar 

  38. Pichon B, Egan D, Rogers M, Gray J. Detection and identification of pathogens and host DNA in unfed host-seeking Ixodes ricinus L. (Acari: Ixodidae). J Med Entomol. 2003;40:723–31.

    CAS  PubMed  Google Scholar 

  39. Assous MV, Wilamowski A, Bercovier H, Marva E. Characterization of relapsing fever. Emerg Infect Dis. 2006;12:1740–3.

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Duscher GG, Leschnik M, Fuehrer HP, Joachim A. Wildlife reservoirs for vector-borne canine, feline and zoonotic infections in Austria. Int J Parasitol Parasites Wildl. 2015;4:88–96.

    PubMed  Google Scholar 

  41. Otranto D, Deplazes P. Zoonotic nematodes of wild carnivores. Int J Parasitol Parasites Wildl. 2019;9:370–83.

    PubMed  PubMed Central  Google Scholar 

  42. Israeli Ministry of Agriculture and Rural Development. https://www.gov.il/he/departments/dynamiccollectors/rabies-occurrence. Accessed 15 June 2023.

  43. International Union for Conservation of Nature (IUCN). 2016. The IUCN Red List of Threatened Species. https://www.iucnredlist.org. Accessed 1 June 2023.

  44. Gallagher J, Clifton-Hadley RS. Tuberculosis in badgers; a review of the disease and its significance for other animals. Res Vet Sci. 2000;69:203–17.

    CAS  PubMed  Google Scholar 

  45. Wodecka B, Michalik J, Lane RS, Nowak-Chmura M, Wierzbicka A. Differential associations of Borrelia species with European badgers (Meles meles) and raccoon dogs (Nyctereutes procyonoides) in western Poland. Ticks Tick Borne Dis. 2016;7:1010–6.

    PubMed  Google Scholar 

  46. Abi-Said MR, Marrouche Abi-Said D. Distribution of the striped hyaena (Hyaena hyaena syriaca Matius, 1882) (Carnivora: Hyaenidae) in urban and rural areas of Lebanon. Zoology of the Middle East. 2007;42:3–14.

    Google Scholar 

  47. Kruuk H. Feeding and social behaviour of the striped hyaena (Hyaena vulgaris Desmarest). Afr J Ecol. 1976;14:91–111.

    Google Scholar 

  48. Hadad E, Kosicki JZ, Yosef R. Population trends of striped hyena (Hyaena hyaena) in Israel for the past five decades. Sci Rep Sci Rep. 2023;13:3982.

    CAS  PubMed  Google Scholar 

  49. Sambu RM, Mathew C, Nonga HE, Lukambagire AS, Yapi RB, Akoko J, et al. Circulating Brucella species in wild animals of the Serengeti ecosystem, Tanzania. One Health Outlook. 2021;3:15.

    CAS  PubMed  PubMed Central  Google Scholar 

  50. Ohiolei JA, Li L, Ebhodaghe F, Yan H, Isaac C, Bo XW, et al. Prevalence and distribution of Echinococcus spp. in wild and domestic animals across Africa: a systematic review and meta-analysis. Transbound Emerg Dis. 2020;67:2345–64.

    CAS  PubMed  Google Scholar 

  51. Drea CM, Coscia EM, Glickman SE. Hyenas. Encyclopedia of Reproduction. New York: Academic Press; 2018. p. 637–45.

    Google Scholar 

  52. Sugianto NA, Buesching CD, Macdonald DW, Newman C. The importance of refining anaesthetic regimes to mitigate adverse effects in very young and very old wild animals: the European badger (Meles meles). J Zool Res. 2019;3:10–7.

    Google Scholar 

  53. Wilamowski A, Assous M, Anis E. Tick-borne relapsing fever (RF) in the civilian population of Israel, 1980–2002. 2005:399–407. In: Fifth International Conference on Urban Pests, Singapore.

  54. Aviles ES, Oakes M, Algranati M, Mansoor AM. Tick-borne relapsing fever. BMJ Case Rep. 2020;13:2018–21.

    Google Scholar 

  55. Shamir M. Antibodies to selected canine pathogens and infestation with intestinal helminths in golden jackals (Canis aureus) in Israel. Vet J. 2001;162:66–72.

    CAS  PubMed  Google Scholar 

  56. Eva B, Bud M. Canine distemper virus in wild carnivore populations from the Czech Republic (2012–2020): Occurrence, geographical distribution, and phylogenetic analysis. Life (Basel). 2022;12:289.

    Google Scholar 

  57. Sykes JE. Immunodeficiencies caused by infectious diseases. Vet Clin North Am Small Anim Pract. 2010;40:409–23.

    PubMed  PubMed Central  Google Scholar 

  58. Barocas A, Hefner R, Ucko M, Merkle JA, Geffen E. Behavioral adaptations of a large carnivore to human activity in an extremely arid landscape. Anim Conserv. 2018;21:433–43.

    Google Scholar 

  59. Hildebrand J, Jurczyk K, Popiołek M, Buńkowska-Gawlik K, Perec-Matysiak A. Occurrence of Borrelia sp. among wild living invasive and native mesocarnivores in Poland. Animals. 2022;12:2829.

    PubMed  PubMed Central  Google Scholar 

  60. Cross ML, Buddle BM, Aldwell FE. The potential of oral vaccines for disease control in wildlife species. Vet J. 2007;174:472–80.

    CAS  PubMed  Google Scholar 

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Acknowledgements

The authors thank Noam Hoffman for her help with handling samples and DNA extraction.

The paper has been sponsored by Elanco Animal Health in the framework of the CVBD® World Forum Symposium

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Authors and Affiliations

  1. Koret School of Veterinary Medicine, Hebrew University of Jerusalem, P.O. Box 12, 7610001, Rehovot, Israel

    Dor Shwartz, Yaarit Nachum-Biala, Stephanie Oren, Kobi Aharoni & Gad Baneth

  2. Pathology Department, Kimron Veterinary Institute, Rishon Lezion, Israel

    Stephanie Oren, Kobi Aharoni, Nir Edery & Lior Moss

  3. Israeli National Parks and Nature Reserves, Jerusalem, Israel

    Roni King & Roi Lapid

  4. Bacteriology and Mycology, Institute for Infectious Diseases and Zoonoses, Ludwig-Maximilians-Universität München, Oberschleißheim, Germany

    Reinhard K. Straubinger

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  1. Dor Shwartz
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Contributions

DS and GB planned and conceived the study and wrote the manuscript. DS and YNB performed the genetic analyses and participated in writing the manuscript. RK and RL collected the carcasses. SO, KA, SA, NE, and LM performed the post-mortem examinations and provided the samples. RKS provided the B. persica culture and participated in writing the manuscript. All authors read and approved the final version of the manuscript.

Corresponding author

Correspondence to Gad Baneth.

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Ethics approval and consent to participate

This study was carried out in accordance with the Hebrew University ethics regulations. The study involved the exclusive use of residual tissue samples taken during post-mortem examination by veterinary pathologists in animals that were killed in road accidents or culled in a rabies protection campaign.

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The authors declare no competing interests.

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Shwartz, D., Nachum-Biala, Y., Oren, S. et al. Borrelia persica infection in wild carnivores in Israel: molecular characterization and new potential reservoirs. Parasites Vectors 16, 337 (2023). https://doi.org/10.1186/s13071-023-05953-4

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Keywords

Parasites & Vectors

ISSN: 1756-3305

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