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Coinfection of Ixodes ricinus (Acari: Ixodidae) in northern Poland with the agents of Lyme borreliosis (LB) and human granulocytic ehrlichiosis (HGE)
Int.J. Med. Microbiol.291 , Suppl.33. 198-201 (2002) © Urban & Fischer Verlag
htlp:/Iw.vw.urbanfischer.de/journalslijmm
original article Coinfection of Ixodes ricinus (Acari: Ixodidae) in northern Poland with the agents of Lyme borreliosis (LB) and human granulocytic ehrlichiosis (HGE) Joanna Stanczak", Maria Racewicz, Wiestawa Kruminis-tozowska, Beata Kubica-Biernat Department of Tropical Parasitology. Institute of Maritime and Tropical Medicine, 9B Powstania Styczniowego St., 81-519 Gdynia, Poland
Abstract Adult Ixodes ricinus ticks were collected from Pomerania province, northern Poland, to determine the presence of infection with agents of human granulocytic ehrlichiosis (HGE) and Lyme borreliosis hy using the polymerase chain reaction (PCR). Of the 424 ticks 19.2 % and 11.6 % contained ehrlichiae and spirochetes, respectively. Frequency of single infection with the HGE agent was 63/424 while frequency of single infection with Borrelia burgdorferi sensu Iato was 28/424. As many as 21/424 ticks (5 % ) contained both pathogens. This finding supports suggestions that both HGE agent and B. burgdorferi 5.1. perpetuate in the same foci and frequently co -infect the same tick vector thereby increasing the risk of humans acquiring mixed infection . .Key words: Borrelia burgdorjeri sensu Iaro - HGE agent - Ixodes ricinus - Lyme borrcliosis, human granulocytic ehrlich iosis - Poland
Introduction Ixodes ricinus, a common and widely distributed tick species in Poland, so far has been mainly known as principal vector of two diseases concerning human health i. e. tick-borne encephalitis (TBE) and Lyme borreliosis (LB). However, preliminary studies (Grzeszczuk et al., 2000; Stanczak et al ., 2000b) have shown that, in our country, I. ricinus can also serve as a vector for another tick-borne infection - human granulocytic ehrlichiosis (HGE), a newly described (Bakken et al., 1996) and potentially deadly disease. Its etiological agent, a bacterium of the genus Ebrlichia, is similar or almost identical (in approx. 98 %) to the veterinary pathogens Ebrlicbia equi and E. phagocytophi!a (Brouqui er al., 1995). Agents of all three diseases (TBE, LB, HGE) can perpetuate in natural cycles between the tick
vector and its vertebrate hosts, sharing common zoonotic reservoirs (Stafford et al ., 1999). The purpose of this study was to estimate the prevalence of HGE agent in I. ricinus collected in the north-eastern region of Poland where B. burgdorferi is widely distributed in tick populations, and provide evidence that these two pathogens can coinfect ticks, thereby raising the possibility of mixed infection in humans.
Materials and methods Adult I. ricinus were collected in May-July 2000 in 12 different forest sites in the Pomerania province, northern Poland, and fixed in 70 % ethanol. Lysatcs were stored in -20°C.
.. Corresponding author: Joanna Stanczak, Institute of Maritime and Tropical Medicine, 9 B Powst. Styczniowego St., 81-519 Gdyn ia Poland, Phone: 0048586998551, Fax: 0048586223354, E-mail: [email protected]}'nia.pl 1438-4221/01/291/8-000
s 15.00/0
I. ricinus coinfection with B. burgdorferi and HGE agent The polymerase chain reaction (PCR) was used to demonstrate the presence of the pathogens in ticks. Borrelial and ehrlichial DNA was extracted from individual specimens by boiling in the presence of NH 40H (Rijpkema et al., 1996). To amplify a 247 bp fragment of 165 r DNA from the HGE agent (Fig. 1), primers EHR 521 (5' TGT AGG CGG TIC GGT AAG TIA AAG 3') and EHR 747 (5' GCA CTC ATC GTI TAC AGC GTG 3') were used (Pancholi et al., 1995). PCR was performed according to Pancholi er al, (1995). In each run, HGE-l strain infected HL60 cells (MRL Diagnostics) were used as a positive control and double distilled water (DDW) as a negative one.
199
Primers FL6 (5' TIC AGG GTC TCA AGC TTG CAC T 3') and FL7 (5' GCA CTC ATC GTT TAC AGC GTG 3') were used to amplify a 276 bp fragment of the flagellin gene of B. burgdorferi s. I. (Picken, 1992) (Fig. 2). PCR was performed as previously described (Stanczak et al., 1999; 2000a). Borrelia burgdorferi sensu stricto strain B 148c12 was used as positive control and DDW and NH 40H as negatives ones. All reactions were carried out in Perkin Elmer GeneAmp PCR System 2400 thermal cyclers. Amplification products were visualised on ethidium bromide-stained 2 % agarose gels.
Results 2
3
4
5
6
7
S
9
10
bp
501
~
4 j).l
::..---
}J I
2<&!
... -
-
-'"-------
-..."
.
-
247 bp
I""
Fig.1. PCR amplification of HGE gene fragment following DNA extraction of unfed Ixodes ricinus ticks. Lane 1: Ml marker; Lanes 2-3: positive control-strain HGE-l; Lane 4: negative control (DDW); Lanes: 5 and 7: negative ticksamples; Lanes 6, 810: positive ticks samples.
I-
.z 3
5
6 7 8 9 10 11
bp ~O l
'£{\.I
3.11
-
2U
276 bp
190
Fig.2. PCR amplification of the {Ia gene of Borrelia burgdorferi sensu lato following extraction of unfed Ixodes ricinus. Lane 1:Ml marker; Lanes 2-3: DDW and NH 40H negative controls; Lane 4: B. burgdorferi strain B-148d2 positive control; Lanes: 5-6 and 9-10: positive ticksamples; Lanes 7-8 and 11: negative ticks samples.
Table 1. Number (percentage) of adult Ixodes ricinus collected in the Pomeranian province (northern Poland), infected with Borrelia burgdorferi sensu latoand HGE agent. Stage
Females Males Total
No. examined
240 184 424
Ticks infected with:
B.burgdorferi
HGE agent
both
neither
19(7.9) 9 (4.9) 28 (6.6)
51 (21.3) 12 (6.5) 63 (14.7)
19(7.9) 2 (1.1) 21 (5.0)
151 (62.9) 161 (87.5) 312 (73.7)
In total, 424 I. ricinus (240 females and 184 males) were examined for the infection with Lyme borreliosis and HGE. Both single species and mixed infections of B. burgdorferi s.I, and human granulocytic Ehrlicbiae were noted (Table I). Frequency of single infection with borreliae was 6.6 %, while with ehrlichiae 14.7%. Frequency of dual infection was 5 %. Overall infection rates were 11.6 % for B. burgdorferi and 19.2 % for the HGE agent. Rates of infection and coinfection were higher in females than in males: app. 2.5 times in the case of B. burgdorferi, 4 times in the case of the agent of HGE and 7 times in the case of both pathogens (Table I). B. burgorferi was detected in 9/12 collection sites while the HGE agent in all but one of them. Percentage of I. ricinus infected with spirochetes varied in particular sites of collection from 0.0 % to 40 % and of those infected with ehrlichiae from 0.0 % to 38.5 % (Table 2).
Discussion The first case of human granulocytic ehrlichiosis in Europe was reported in 1997 from Slovenia (Petrovec et al., 1997). In the same year, the presence of its causative agent in European ticks was demonstrated in Sweden when it was detected in 6.6 % of 151 tested Ixodes ricinus nymphs (von Stedingk et aI., 1997). In 1998, investigation was extended to adults and 5.4 % (25/462) tested were found to be positive. Moreover, Parola et al. (l998) detected HGE agent in 1.3 % (1/80) of examined ticks in France. Results of our study demonstrate that the agent of HGE is frequently noted also in I. ricinus in Poland and that ehrlichiae and B. burgdor[eri perpetuate in the tick populations previously identified as Lyme borreliosis foci in the northern region of our country.
200
J. Stanczak et al.
The dual infection of Ixodes TlCIIIllS with both pathogens noted by us seems to be frequent and confirms findings of other authors. For instance, in the USA Schauber et al, (1998) detected mixed infection in 28.2 % (53/188) of adult I. scapularis collected in New York state, while Levin et al. (1999) in 14 % (4/100) of those collected in Connecticut. Higher infection level of ticks with HGE agent (up to 38.8 %) than with B: burgdorferi suggests that the first one was prevalent in the area studied. Physicians in this area should be aware of the risk for HGE, in particular, because first human cases have been already recorded in Poland (Tylewska-Wierzbanowska et al., 2000). Moreover, possibility of double infection should be considered for patients who declared a tick bite and/or developed Lyme borreliosis or flu-like symptoms. Mitchel et al. (1996) noted that in Minnesota and Wisconsin, 5.2 % (5/96) of patients with Lyme borreliosis demonstrated immunoserologic evidence of coinfection with the agent of HGE and 5.3 % (1/19) patients diagnosed with HGE showed coinfection with B. bllrgdorferi.·Also Mazella et al. (1996) reported a case of concurrent presentation of human ehrlichiosis and Lyme disease in Connecticut. 'Further, extensive investigations are needed. Acknowledgements. This research was supported financially by Grant No.6 P04C 047 17 from the State Committee for scientific Research, Warsaw, Poland.
References Bakken, J. 5., Krueth, J., Tilden, R. L., Dumler,]. 5., Kristiansen, B. E.: Serological evidence of human granulocytis ehrlichiosis in Norway. Eur.], C1in. Microbiol. Infect. Dis. 15, 829-832 (1996). Brouqui, P., Dumler, j. 5., Lienhard, R., Brossard, M., Raoulr, D.: Human granulocytic ehrlichiosis in Europe. Lancet 346, 782-783 (1995). Grzeszczuk, A., Stanczak, J., Kubica-Biernat, B., KruminisLozowska, \Y/., Racewicz, M.: First seroepidemiological evidence of human granulocytic ehrlichiosis (HGE) in Poland. Preliminary results. Abstracts VIII European Multicolloquium of Parasitology. Acta Parasitol.45, 219 (2000). Levin, M. L., des Vignes, E, Fish, D.: Disparity in the natural cycles of Borrelia burgdorferi and the agent of human granulocytic ehrlichiosis. Emerg. Infect. Dis. 5, 204-208 (1999). : Mazzella, EM., Roman, A., Perez, A.: A case of concurrent presentation of human ehrlichiosis and Lyme disease in Connecticut. Conn. Ivied: 60, 515-519 (1996). Mitchel, P.D., Reed, K. D., Hofkes, ]. M.: Immunoserologic evidence of coinfection with Borrelia burgdorferi, Babesia microti, and human granulocytic Ebrlicbia species in residents of Wisconsin and Minnesota. J. Clin. Microbiol. 34, 724-727 (1996).
Table 2. Prevalence of infection with Borrelia burgdorferi s.1. and the HGE agent among adultIxodes ricinus ticks collected in the Pomeranian province (northern Poland). Study site
Ixodes ricinus no. tested
no./% Bb1 infected
no./% HGE agent infected
1.Krynica Morska 2.Babi 061 3.Sianowo 4.Borucino 5.Babid61 k, Kolbud 6.Wieiyca 7.Buszkowy 8. Parkwice 9.Wejherowo 10.Brzyno 11.Godetowo 12.BI~dno
35 22 14 109 13 15 9 40 16 22 34 39
4/11.4 0/0.0 117.1 10/9.2 2/15.4 6/40.0 0/0.0 8/20.0 116.3 5/22.5 0/0.0 9/23.1
4/25.0 7/31.2 5/35.7 26/23.9 117.7 0/0.0 2/22.2 12/30.0 3/18.8 4118.8 3/8.8 15/38.5
Total
424
49/11.6
84/19.2
I
Bb - Borrelia burgdorferi sensu lato.
Pancholi, P., Kolberr, C. P., Mitchel, P.D., Reed, K. D., Dumler,]. 5., Bakken, J. 5., Telford, S. R. III, Persing, D.: Ixodes dammini as a potential vector of human granulocytic ehrlichiosis.].lnfect. Dis. 172, 1007-1012 (1995). Parola, P., Beati, L., Cam bon, M., Brouqui, P., Raoult, D.: Ehrlichial DNA amplified from Ixodes ricinus (Acari: Ixodidae) in France.]. Med. Entomol. 35,180-183 (1998). Petrovec, M., Lotric Furlan,S., Avsic Zupanc, T., Strle, E, Brouqui, P., Roux, V., Dumler,]. 5.: Human' disease in Europe caused by a granulocytic ehrlichia species.]. Clin. Microbiol. 35,1556-1559 (1997). Picken, R. N.: Polymerase chain reaction primers and probes derived from flagellin gene sequences for specific detection of the agents of Lyme disease and North American relapsing fever.], Clin. Microbiol. 30, 99-114 (1992). Rijpkerna, 5., Golubi', D., Molkenboer, M., Verbreek-De Kruif, N., Schellekens,].: Identification of four groups of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected in a Lyme borreliosis endemic region of northern Croatia. Exp. App. Acarol. 20, 23-30 (1996). Schauber, E. M., Gertz, S.J., Maple, wr, Ostfeld, R. 5.: Coinfection of blacklegged ticks (Acari: Ixodidae) in Dutchess County, New York, with the agents of Lyme disease and human granulocytic ehrlichiosis.J. Med. Entomol. 35,901-903 (1998). Stafford, K. C. 3,d, Massung, R. E, Magnarelli, L. A., Ijdo, J. W., Anderson,]. E: Infection with agents of human granulocytic ehrlichiosis, Lyme disease, and babesiosis in wild white-footed mice tl'eromyscus leucopust in Connecticut.]. Clin. Microbiol. 37,2887-2892 (1999). Stanczak, J., Racewicz, M., Kubica-Biernat, B., KrurninisLozowska, W., DAbrowski, J., Adamczyk, A., Markowska, M.: Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks (Acari, Ixodidae) in different Polish woodlands. Ann. Agric. Environ. Med. 6, 127-132 (1999).
I. ricinus coinfection with B. burgdorfcn and HGE agent Stanczak, J., Kubica-Biernat, B., Raccwicz, 10.1., KrurninisLozowska, W., Kur, J.: Detection of three gcnospccies of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks in different regions of Poland. Int.]' Med. Microbiol. 290, 559-566 (2000a). Stanczak, ]., Raccwicz, M ., Kubica-Biernat, B., KrurninisLozowska, W.: Detection of the agent of human granulocytic ehrlichiosis (HGE) in ticks in Poland. Abstracts VIII European Multicolloquium of Parasitology. Acta Parasitol. 45, 217 (2000b).
201
Tylewska-Wierzbanowska, S., Chmielewski, T., Kondrusik, 1'.1 ., Hermanowska-Szpakowicz, T., Sawicki, W., Sulrek, K.: Three acute cases of human granulO Cytic ehrl ichiosis rcconized in Poland. Clin . M icrobiol. Infect. 6 Suppl. 1, 195 (2000). von Sredingk, L. V., Giirtelschmid, 1\1., Hanson, H .S ., Gustafson, R., Dotevall, L., Engvall, E.O., Grandstrom, M .: The human granulocytic ehrlichiosis (HGE) agent in Swedish ticks. C1in. Infect. Dis. 3,573-574 (1997) .
htlp:/Iw.vw.urbanfischer.de/journalslijmm
original article Coinfection of Ixodes ricinus (Acari: Ixodidae) in northern Poland with the agents of Lyme borreliosis (LB) and human granulocytic ehrlichiosis (HGE) Joanna Stanczak", Maria Racewicz, Wiestawa Kruminis-tozowska, Beata Kubica-Biernat Department of Tropical Parasitology. Institute of Maritime and Tropical Medicine, 9B Powstania Styczniowego St., 81-519 Gdynia, Poland
Abstract Adult Ixodes ricinus ticks were collected from Pomerania province, northern Poland, to determine the presence of infection with agents of human granulocytic ehrlichiosis (HGE) and Lyme borreliosis hy using the polymerase chain reaction (PCR). Of the 424 ticks 19.2 % and 11.6 % contained ehrlichiae and spirochetes, respectively. Frequency of single infection with the HGE agent was 63/424 while frequency of single infection with Borrelia burgdorferi sensu Iato was 28/424. As many as 21/424 ticks (5 % ) contained both pathogens. This finding supports suggestions that both HGE agent and B. burgdorferi 5.1. perpetuate in the same foci and frequently co -infect the same tick vector thereby increasing the risk of humans acquiring mixed infection . .Key words: Borrelia burgdorjeri sensu Iaro - HGE agent - Ixodes ricinus - Lyme borrcliosis, human granulocytic ehrlich iosis - Poland
Introduction Ixodes ricinus, a common and widely distributed tick species in Poland, so far has been mainly known as principal vector of two diseases concerning human health i. e. tick-borne encephalitis (TBE) and Lyme borreliosis (LB). However, preliminary studies (Grzeszczuk et al., 2000; Stanczak et al ., 2000b) have shown that, in our country, I. ricinus can also serve as a vector for another tick-borne infection - human granulocytic ehrlichiosis (HGE), a newly described (Bakken et al., 1996) and potentially deadly disease. Its etiological agent, a bacterium of the genus Ebrlichia, is similar or almost identical (in approx. 98 %) to the veterinary pathogens Ebrlicbia equi and E. phagocytophi!a (Brouqui er al., 1995). Agents of all three diseases (TBE, LB, HGE) can perpetuate in natural cycles between the tick
vector and its vertebrate hosts, sharing common zoonotic reservoirs (Stafford et al ., 1999). The purpose of this study was to estimate the prevalence of HGE agent in I. ricinus collected in the north-eastern region of Poland where B. burgdorferi is widely distributed in tick populations, and provide evidence that these two pathogens can coinfect ticks, thereby raising the possibility of mixed infection in humans.
Materials and methods Adult I. ricinus were collected in May-July 2000 in 12 different forest sites in the Pomerania province, northern Poland, and fixed in 70 % ethanol. Lysatcs were stored in -20°C.
.. Corresponding author: Joanna Stanczak, Institute of Maritime and Tropical Medicine, 9 B Powst. Styczniowego St., 81-519 Gdyn ia Poland, Phone: 0048586998551, Fax: 0048586223354, E-mail: [email protected]}'nia.pl 1438-4221/01/291/8-000
s 15.00/0
I. ricinus coinfection with B. burgdorferi and HGE agent The polymerase chain reaction (PCR) was used to demonstrate the presence of the pathogens in ticks. Borrelial and ehrlichial DNA was extracted from individual specimens by boiling in the presence of NH 40H (Rijpkema et al., 1996). To amplify a 247 bp fragment of 165 r DNA from the HGE agent (Fig. 1), primers EHR 521 (5' TGT AGG CGG TIC GGT AAG TIA AAG 3') and EHR 747 (5' GCA CTC ATC GTI TAC AGC GTG 3') were used (Pancholi et al., 1995). PCR was performed according to Pancholi er al, (1995). In each run, HGE-l strain infected HL60 cells (MRL Diagnostics) were used as a positive control and double distilled water (DDW) as a negative one.
199
Primers FL6 (5' TIC AGG GTC TCA AGC TTG CAC T 3') and FL7 (5' GCA CTC ATC GTT TAC AGC GTG 3') were used to amplify a 276 bp fragment of the flagellin gene of B. burgdorferi s. I. (Picken, 1992) (Fig. 2). PCR was performed as previously described (Stanczak et al., 1999; 2000a). Borrelia burgdorferi sensu stricto strain B 148c12 was used as positive control and DDW and NH 40H as negatives ones. All reactions were carried out in Perkin Elmer GeneAmp PCR System 2400 thermal cyclers. Amplification products were visualised on ethidium bromide-stained 2 % agarose gels.
Results 2
3
4
5
6
7
S
9
10
bp
501
~
4 j).l
::..---
}J I
2<&!
... -
-
-'"-------
-..."
.
-
247 bp
I""
Fig.1. PCR amplification of HGE gene fragment following DNA extraction of unfed Ixodes ricinus ticks. Lane 1: Ml marker; Lanes 2-3: positive control-strain HGE-l; Lane 4: negative control (DDW); Lanes: 5 and 7: negative ticksamples; Lanes 6, 810: positive ticks samples.
I-
.z 3
5
6 7 8 9 10 11
bp ~O l
'£{\.I
3.11
-
2U
276 bp
190
Fig.2. PCR amplification of the {Ia gene of Borrelia burgdorferi sensu lato following extraction of unfed Ixodes ricinus. Lane 1:Ml marker; Lanes 2-3: DDW and NH 40H negative controls; Lane 4: B. burgdorferi strain B-148d2 positive control; Lanes: 5-6 and 9-10: positive ticksamples; Lanes 7-8 and 11: negative ticks samples.
Table 1. Number (percentage) of adult Ixodes ricinus collected in the Pomeranian province (northern Poland), infected with Borrelia burgdorferi sensu latoand HGE agent. Stage
Females Males Total
No. examined
240 184 424
Ticks infected with:
B.burgdorferi
HGE agent
both
neither
19(7.9) 9 (4.9) 28 (6.6)
51 (21.3) 12 (6.5) 63 (14.7)
19(7.9) 2 (1.1) 21 (5.0)
151 (62.9) 161 (87.5) 312 (73.7)
In total, 424 I. ricinus (240 females and 184 males) were examined for the infection with Lyme borreliosis and HGE. Both single species and mixed infections of B. burgdorferi s.I, and human granulocytic Ehrlicbiae were noted (Table I). Frequency of single infection with borreliae was 6.6 %, while with ehrlichiae 14.7%. Frequency of dual infection was 5 %. Overall infection rates were 11.6 % for B. burgdorferi and 19.2 % for the HGE agent. Rates of infection and coinfection were higher in females than in males: app. 2.5 times in the case of B. burgdorferi, 4 times in the case of the agent of HGE and 7 times in the case of both pathogens (Table I). B. burgorferi was detected in 9/12 collection sites while the HGE agent in all but one of them. Percentage of I. ricinus infected with spirochetes varied in particular sites of collection from 0.0 % to 40 % and of those infected with ehrlichiae from 0.0 % to 38.5 % (Table 2).
Discussion The first case of human granulocytic ehrlichiosis in Europe was reported in 1997 from Slovenia (Petrovec et al., 1997). In the same year, the presence of its causative agent in European ticks was demonstrated in Sweden when it was detected in 6.6 % of 151 tested Ixodes ricinus nymphs (von Stedingk et aI., 1997). In 1998, investigation was extended to adults and 5.4 % (25/462) tested were found to be positive. Moreover, Parola et al. (l998) detected HGE agent in 1.3 % (1/80) of examined ticks in France. Results of our study demonstrate that the agent of HGE is frequently noted also in I. ricinus in Poland and that ehrlichiae and B. burgdor[eri perpetuate in the tick populations previously identified as Lyme borreliosis foci in the northern region of our country.
200
J. Stanczak et al.
The dual infection of Ixodes TlCIIIllS with both pathogens noted by us seems to be frequent and confirms findings of other authors. For instance, in the USA Schauber et al, (1998) detected mixed infection in 28.2 % (53/188) of adult I. scapularis collected in New York state, while Levin et al. (1999) in 14 % (4/100) of those collected in Connecticut. Higher infection level of ticks with HGE agent (up to 38.8 %) than with B: burgdorferi suggests that the first one was prevalent in the area studied. Physicians in this area should be aware of the risk for HGE, in particular, because first human cases have been already recorded in Poland (Tylewska-Wierzbanowska et al., 2000). Moreover, possibility of double infection should be considered for patients who declared a tick bite and/or developed Lyme borreliosis or flu-like symptoms. Mitchel et al. (1996) noted that in Minnesota and Wisconsin, 5.2 % (5/96) of patients with Lyme borreliosis demonstrated immunoserologic evidence of coinfection with the agent of HGE and 5.3 % (1/19) patients diagnosed with HGE showed coinfection with B. bllrgdorferi.·Also Mazella et al. (1996) reported a case of concurrent presentation of human ehrlichiosis and Lyme disease in Connecticut. 'Further, extensive investigations are needed. Acknowledgements. This research was supported financially by Grant No.6 P04C 047 17 from the State Committee for scientific Research, Warsaw, Poland.
References Bakken, J. 5., Krueth, J., Tilden, R. L., Dumler,]. 5., Kristiansen, B. E.: Serological evidence of human granulocytis ehrlichiosis in Norway. Eur.], C1in. Microbiol. Infect. Dis. 15, 829-832 (1996). Brouqui, P., Dumler, j. 5., Lienhard, R., Brossard, M., Raoulr, D.: Human granulocytic ehrlichiosis in Europe. Lancet 346, 782-783 (1995). Grzeszczuk, A., Stanczak, J., Kubica-Biernat, B., KruminisLozowska, \Y/., Racewicz, M.: First seroepidemiological evidence of human granulocytic ehrlichiosis (HGE) in Poland. Preliminary results. Abstracts VIII European Multicolloquium of Parasitology. Acta Parasitol.45, 219 (2000). Levin, M. L., des Vignes, E, Fish, D.: Disparity in the natural cycles of Borrelia burgdorferi and the agent of human granulocytic ehrlichiosis. Emerg. Infect. Dis. 5, 204-208 (1999). : Mazzella, EM., Roman, A., Perez, A.: A case of concurrent presentation of human ehrlichiosis and Lyme disease in Connecticut. Conn. Ivied: 60, 515-519 (1996). Mitchel, P.D., Reed, K. D., Hofkes, ]. M.: Immunoserologic evidence of coinfection with Borrelia burgdorferi, Babesia microti, and human granulocytic Ebrlicbia species in residents of Wisconsin and Minnesota. J. Clin. Microbiol. 34, 724-727 (1996).
Table 2. Prevalence of infection with Borrelia burgdorferi s.1. and the HGE agent among adultIxodes ricinus ticks collected in the Pomeranian province (northern Poland). Study site
Ixodes ricinus no. tested
no./% Bb1 infected
no./% HGE agent infected
1.Krynica Morska 2.Babi 061 3.Sianowo 4.Borucino 5.Babid61 k, Kolbud 6.Wieiyca 7.Buszkowy 8. Parkwice 9.Wejherowo 10.Brzyno 11.Godetowo 12.BI~dno
35 22 14 109 13 15 9 40 16 22 34 39
4/11.4 0/0.0 117.1 10/9.2 2/15.4 6/40.0 0/0.0 8/20.0 116.3 5/22.5 0/0.0 9/23.1
4/25.0 7/31.2 5/35.7 26/23.9 117.7 0/0.0 2/22.2 12/30.0 3/18.8 4118.8 3/8.8 15/38.5
Total
424
49/11.6
84/19.2
I
Bb - Borrelia burgdorferi sensu lato.
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