Seroprevalence of Bartonella Species in Patients with Ocular Inflammation.

Bartonella species, vector-borne etiologic agents of many systemic or self-limited infections, are responsible for a widening spectrum of diseases in humans, including inflammatory conditions of the eye. The aim of this study was to determine whether there is any relationship between uveitis and the evidence of Bartonella spp. infection in the serum, ocular fluid, and cataract mass in patients with intraocular inflammation. Polymerase chain reaction (PCR)-based tests and DNA sequencing were performed on surgery-extracted specimens of intraocular fluid and lens mass of 33 patients. Sera from 51 patients and 101 control subjects were tested for the presence of specific antibodies against Bartonella spp. Neither IgM-class antibodies against Bartonella spp. nor Bartonella spp. DNA were detected. A specific IgG-class antibody was found in 33.3% of the patients with uveitis. The rate of positive Bartonella serology was higher among the uveitis patients than that in control subjects. This high rate may in part result from unrecognized indirect mechanisms rather than the immediate presence and multiplication of Bartonella spp. in the eyeball. Nonetheless we believe that screening for Bartonella spp. should become part of the diagnostic workup in uveitis.

Bactericidal Activity of Octenidine to Various Genospecies of Borrelia burgdorferi, Sensu Lato Spirochetes in Vitro and in Vivo.

The aim of our studies was to invent a reliable method for detection of the bactericidal activity of disinfectants against Borrelia burgdorferi in suspension (in vitro) and in cell line cultures (in vivo). In the suspension method, 0.01% octenidine at 20°C and 35°C was bactericidal to Borrelia afzeli; Borrelia garini, B. burgdorferi sensu stricto after 5 minutes treatment. Increase of the temperature to 35°C speed up the bactericidal effect to 1 minute. The bactericidal action of octenidine towards B. burgdorferi spirochetes growing in fibroblasts was less effective and needed a longer time to kill them than in the suspension.

Borrelia miyamotoi - new etiologic agent of neuroborreliosis?

Borrelia miyamotoi spirochetes discovered in Ixodes persulcatus ticks in Japan, in 1994 and documented in ticks and rodents in moderate climate zone of northern hemisphere. They belong to tick-borne relapsing fever group spirochetes. Borrelia miyamotoi is an etiologic agent of B.miyamotoi disease with acute febrile illness, including fever, headache, dizziness, fatigue, chills, and muscle and joint pain. Recurrence of fever has been observer in more than 10% patients. In some patients meningoencephalitis, encephalitis and cranial neuritis were observed. Laboratory recognition is based mainly upon PCR testing. Serological testing is limited due to inaccessibility of tests.

Analysis of nanomechanical properties of Borrelia burgdorferi spirochetes under the influence of lytic factors in an in vitro model using atomic force microscopy.

BACKGROUND: Atomic force microscopy (AFM) is an experimental technique which recently has been used in biology, microbiology, and medicine to investigate the topography of surfaces and in the evaluation of mechanical properties of cells. The aim of this study was to evaluate the influence of the complement system and specific anti-Borrelia antibodies in in vitro conditions on the modification of nanomechanical features of B. burgdorferi B31 cells. MATERIAL AND METHODS: In order to assess the influence of the complement system and anti-Borrelia antibodies on B. burgdorferi s.s. B31 spirochetes, the bacteria were incubated together with plasma of identified status. The samples were applied on the surface of mica disks. Young's modulus and adhesive forces were analyzed with a NanoScope V, MultiMode 8 AFM microscope (Bruker) by the PeakForce QNM technique in air using NanoScope Analysis 1.40 software (Bruker). RESULTS/CONCLUSION: The average value of flexibility of spirochetes' surface expressed by Young's modulus was 10185.32 MPa, whereas the adhesion force was 3.68 nN. AFM is a modern tool with a broad spectrum of observational and measurement abilities. Young's modulus and the adhesion force can be treated as parameters in the evaluation of intensity and changes which take place in pathogenic microorganisms under the influence of various lytic factors. The visualization of the changes in association with nanomechanical features provides a realistic portrayal of the lytic abilities of the elements of the innate and adaptive human immune system.

Bacterial tick-borne diseases caused by Bartonella spp., Borrelia burgdorferi sensu lato, Coxiella burnetii, and Rickettsia spp. among patients with cataract surgery.

BACKGROUND: Clinical data have shown that tick-borne diseases caused by Borrelia burgdorferi sensu lato, Bartonella spp., Coxiella burnetii, and Rickettsia spp. can affect the central nervous system, including the eye. The aim of this study was to establish a relationship between the incidence of cataract and evidence of bacterial infections transmitted by ticks. MATERIAL AND METHODS: Fluid with lenticular masses from inside of the eye and blood from 109 patients were tested by PCR and sequencing. Sera from patients and the control group were subjected to serological tests to search specific antibodies to the bacteria. RESULTS: Microbiological analysis revealed the presence of Bartonella sp. DNA in intraoperative specimens from the eye in 1.8% of patients. Serological studies have shown that infections caused by B. burgdorferi sensu lato and Bartonella sp. were detected in 34.8% and 4.6% of patients with cataract surgery, respectively. CONCLUSIONS: Presence of DNA of yet uncultured and undescribed species of Bartonella in eye liquid indicates past infection with this pathogen. Specific antibodies to B. burgdorferi sensu lato and Bartonella sp. are detected more frequently in patients with cataract compared to the control group. This could indicate a possible role of these organisms in the pathological processes within the eyeball, leading to changes in the lens. Further studies are needed to identify Bartonella species, as well as to recognize the infectious mechanisms involved in cataract development.

Q fever outbreaks in Poland during 2005-2011.

BACKGROUND: Q fever is a health problem affecting humans and animals worldwide. In Poland, previous studies have pointed to 2 sources of outbreaks of the disease: the importation of infected animals and their products, and natural domestic foci. In the last decade, 5 outbreaks have occurred in cattle farms in south Poland in Malopolskie, Podkarpackie, Opolskie, and Silesian provinces. The aim of this study was to characterize the Q fever foci in Poland. MATERIAL AND METHODS: A total of 279 individuals were included. Levels of serum IgM and IgG antibodies to phase I and II C. burnetii antigens were assayed by indirect immunofluorescence method. Bacterial DNA from all specimens were detected with PCR with primer pairs specific to the htpAB-associated repetitive element, and amplicons were sequenced. RESULTS: Infection was recognized in 67 individuals out of 279 tested in all foci. Twenty-five individuals presented clinical symptoms of acute Q fever. DNA of C. burnetii was found in 8 human blood samples obtained from 3 farm workers and 5 family members. CONCLUSIONS: The described outbreaks demonstrate that the main source of human infections in Poland is infected cattle.

Prevalence of rickettsioses in Poland in 2006-2012.

INTRODUCTION: Tick-borne diseases, such as rickettsioses, anaplasmosis, Lyme boreliosis and bartonellosis are often difficult to correctly diagnose. All these disease are present in Poland. OBJECTIVES: The aim of the study was to estimate a prevalence of Rickettsia spp. infections in humans in Poland in 2006 to 2012 based on the results made in the Laboratory of Rickettsiae, Chlamydiae and Spirochetes, NIPH-NIH in Warsaw. MATERIAL AND METHODS: The levels of Rickettsia spp. and Anaplasma phagocytophilum IgM and IgG antibodies were determined by indirect immunofluorescent assay (IFA). From 2006 to 2012, serum samples derived from 180 humans suspected for rickettsioses, including 84 patients suspected for the infections with typhus and spotted fever group (SFG) rickettsiae, and 96 patients suspected for anaplasmosis. RESULTS: Specific serum antibodies to the SFG rickettsiae have been detected in 5 persons (2.7%). Granulocytic anaplasmosis has been recognized in 9 patients (4.9%). While the reporting and registration of rickettsioses are obligatory in Poland less than 50% of detected cases are reported. CONCLUSIONS: Presented date indicate that in Poland rickettsioses are often unrecognized resulting in their underestimation. If research for rickettsiosis are made immediately after infection, antibodies will not be detect.

Characteristics of Bartonella spp. infections in Poland in the years 2009-2012 identified in the laboratory of National Institute of Public Health--National Institute of Hygiene.

INTRODUCTION: Various Bartonella species, (Gram-negative aerobic bacilli) are etiologic agents ofzoonotic diseases called bartonelloses, which manifest with different symptoms depending on the bacterial species, reservoir and vector. In Poland and Europe, the most common bacterial species of the genus Bartonella is Bartonella henselae. MATERIAL AND METHODS: [corrected] Serum samples derived from patients with clinical symptoms suggesting Bartonella spp. infection, sent in 2009-2012 to the Laboratory of Rickettsiae, Chlamydiae and Spirochaetes of National Institute of Public Health--National Institute of Hygiene in Warsaw were tested. Levels of specific IgM and IgG antibodies to B. henselae and B. quintana antigens were detected with indirect immunofluorescence method (IFA). RESULTS: Six hundred sixty three serum samples were examined from humans with clinical symptoms suggestive bartonellosis, in 2009-2012. Specific antibodies for B. henselae were detected in 435 patients (65.6%). IgM antibodies were found in 93 patients (21.4%) including 11 patients (2.5%) with IgM only. IgG antibodies were identified in 424 people (78.6%) of whom 342 had IgG antibodies only. The antibodies of both classes were detected in 82 people (18.9%). B. quintana infections were not found. The majority of samples for study of bartonellosis were submitted in the autumn. In patients with confirmed bartonellosis, the most common symptoms of disease were lymphadenopathy (86 people, 13%), fever (13 patients, 2%) and nodular changes in various organs (13 patients, 2%). CONCLUSIONS: Infections caused by Bartonella spp. in Poland should be monitored to acquire the information on the frequency and distribution of disease in the country and their clinical course.

Case report of African tick-bite fever from Poland.

A confirmed case of rickettsiosis acquired in South Africa and recognized in Poland was described. The patient fulfilled clinical criteria highly suggestive of African tick bite fever, such as eschars, regional lymphadenitis, cutaneous rash within 10 days after his return from sub-Saharan Africa. Infection with Rickettsia africae was confirmed by polymerase chain reaction and sequencing.

Q fever at the turn of the century.

Q fever is an infectious zoonotic disease characterized by sudden fever, headache, and atypical pneumonia, caused by Coxiella burneti--an obligatory intracellular parasite. Based on phylogenetic analysis of the genes sequences, the classification was changed and C. burnetii species was included to the gamma subgroup of the proteobacteria, Legionellales order and Coxiellaceae family. This analysis showed more than 99% sequence similarity of 16SrRNA gene among the strains isolated in different regions of the world. Q fever is a widespread in the world zoonosis. Its main reservoir in the rural environment are farm animals: cows, sheep, goats, and urban pets such as dogs, cats, rabbits. In acute infection these bacteria are detected in various internal organs such as lungs, liver, spleen, and in excretion in urine, faeces and milk. During childbirth, they occur in large number in the amniotic fluid and placenta. Recently, it has been found that free-living amoeba Acanthamoeba castellani may also be a reservoir of the pathogen. The intra-amoebal location of C. burnetii cells was observed.

Susceptibility of Polish Bartonella henselae strains.

Due to the fastidious nature of B. henselae and the limited number of available isolates worldwide, there are few data on its in vitro susceptibility to antibiotics. We determined the minimal inhibitory concentrations (MIC) of ten antimicrobial agents against 11 feline isolates of B. henselae by Etest method. The lowest MICs were obtained for rifampicin < or = 0.002 mg/L. MICs of all isolates were < 0.016 mg/L for ampicilin, amoxicillin/clavulanic acid, tetracycline and ranged from 0.016 to 0.032 mg/L for azithromycin. The MICs for two tested fluoroquinolones: ciprofloxacin and levofloxacin ranged from 0.016 to 0.125 mg/L. The highest MICs were obtained for gentamicin ranging from 0.025 to 2.0 mg/L. Sulphonamide resistance genes sul 1, sul 2, sul 3 were not found in any of the tested isolates. Etest methodology seems to be a reliable method for determination of B. henselae susceptibility, however standardization is strongly desired.

[Leptospira spp. and Coxiella burnetii infections occuring in Radomskie District in people of selected professional groups]. / Zakazenia Leptospira spp. i Coxiella burnetii wystepujace w powiecie radomskim u ludzi z wybranych grup zawodowych.

INTRODUCTION: Leptospirosis and Q fever are a zoonotic diseases with global occurring. OBJECTIVES: The aim of this study was to determine the prevalence of Leptospira spp. and C. burnetii in humans, who have contacts with infected animals or are exposed to an environment potentially contaminated with these bacteria. MATERIAL AND METHODS. Blood serum samples originating from 177 veterinarians and farmers and 134 garbage collectors (blood samples) were examined. Control group consisted 43 blood samples derived from blood donors. For the detection of specific IgM and IgG antibodies of Leptospira spp., enzyme-linked immunosorbent assay (ELISA) was used. Indirect immunofluorescence method (IFA) was used for detection of specific IgG C. burnetii antibodies. DNA of Leptospira spp. and C. burnetii was detected by PCR method with appropriate pairs of primers. RESULTS: Specific IgG C. burnetii antibodies of phase II were detected in sera of 4.4% of the farmers and veterinarians, and in 12% of garbage men. Antibodies in blood donors was not found. Antibodies of Leptospira spp. were present in the serum of 23.6% of farmers and veterinarians, 26.2% of garbage men and 14% of blood donors. C. burnetii DNA was detected in one sample derived from the veterinarian (1.1%). Leptospira spp. DNA was not detected in tested material. Blood samples from farmers, veterinarians and garbage collectors showed the higher prevalence of antibodies of Leptospira spp. and C. burnetii as compared to the control group (blood donors). CONCLUSIONS: Beside farmers and veterinarians, garbage collectors should be consider as high risk group of contracting leptospirosis and Q fever. Both leptospirosis as well as Q fever should be considered in the differential diagnosis in humans with animals and animals' material contact when they reveal flu-like symptoms.

Spotted fever rickettsiosis caused by Rickettsia raoultii--case report.

We report the case of rickettsial eschar-associated spotted fever, most probable due to Rickettsia raoultii, an emerging pathogen, which was previously described in patients with tick-borne lymphadenopathy (TIBOLA), also called Dermacentor-borne necrosis erythema and lymphadenopathy (DEBONEL). The pathogenicity of R. raoultii is not well established. The survey of ticks from Poland (Ixodes ricinus and Dermacentor reticulatus) revealed that R. raoultii occur in all regions of Poland and predominate over other rickettsiae of spotted fever group--R. slovaca and R. helvetica. A 17-year-old otherwise healthy girl was admitted to Department of Zoonotic and Tropical Diseases because of fever, eschar and rash. Multiple disseminated small lesions were present on the skin of her head, trunk and limbs, also palms and soles, and mucosa of her lips. The majority of them had necrotic center slightly elevated with redness around, single ones had vesicular appearance. The lymph nodes on the right side of her neck were enlarged. Laboratory investigations revealed: leukopenia (with 22% of bands in differential), thromocytopenia, slightly elevated C-Reactive Protein, as well as procalcytonin. The quick improvement was observed with a treatment with ceftriaxone and doxycycline. Two weeks after the onset of disease, IgG serum antibodies titer of 128 reacting with R. rickettsii antigen only was detected. IFA tests with six SFG rickettsial species demonstrated the strongest reaction with R. raoultii group antigens in a titer of 64. The case we report, resembling boutonneuse fever, with leukopenia, thrombocytopenia and septic parameters indicates possible higher virulence of R. raoultii than it was previously observed.

Inhibition of fibroblast apoptosis by Borrelia afzelii, Coxiella burnetii and Bartonella henselae.

Apoptosis is a genetically controlled mechanism of cell death involved in the regulation of tissue homeostasis. The aim of this study was to investigate the influence of Borrelia afzelii, Coxiella burnetii, and Bartonella henselae bacteria on apoptosis measured as the level of caspase 3 activity in human fibroblast cells HEL-299. Our findings show that C. burnetii bacteria may inhibit the process of apoptosis in the host cells for a long time. This can permit intracellular survival in the host and mediatingthe development of chronic disease.

Tick-borne infections as a cause of heart transplantation.

Many bacterial species can be a cause of various heart diseases, such as: Borrelia burgdorferi sensu lato, Coxiella burnetii and Bartonella spp. The aim of the present studies was to establish if any tick-borne infections can contribute to serious heart disorders resulting in the need for heart transplantation. Myocardium, aortic and mitral valve samples from hearts removed from patients undergoing heart transplantation were tested. The presence of Bartonella spp., Borrelia afzeli and C. burnetii bacteria in malfunctioning human hearts has been shown. DNA of Bartonella spp., B. burgdorferi and C. burnetii were detected in various parts of tested hearts. DNA of B. afzelii and Bartonella spp. were found in the aortic valves. DNA of C. burnetii was detected in the myocardium. Mixed infections with Bartonella spp. and C. burnetii were also observed. Obtained results indicate that diagnosis of Bartonella spp., B. burgdorferi C. burnetii and Rickettsia spp. infections should be considered in cases of infectious endocarditis with negative blood cultures.

[Case of TIBOLA/DEBONEL (tick - borne lymphadenopathy/Dermacentor spp. - borne necrosis - erythema - lymphadenopathy) in Poland]. / Przypadek TIBOLA/DEBONEL (tick - borne lymphadenopathy/Dermacentor spp. - borne necrosis - erythema - lymphadenopathy) w Polsce.

Rickettsia slovaca was recognized as the causative agent of the disease named tick - borne lymphadenopathy (TIBOLA) or Dermacentor spp.- borne necrosis - erythema lymphadenopathy (DEBONEL). Generally, this ricketsiosis is considered a mild disease but the disease is considered a mild rickettsiosis, but severe symptoms can appeare, especially in untreated patients. Described infection is the first case of TIBOLA/DEBONEL found in Poland. The patient had typical symptoms, such as enlarged painful cervical lymph nodes and maculo - papular rash. Milium on the scalp behind the ear concha, was a probably scar formation associated with typical eschar. The diagnosis was confirmed by detection of 128 titer in MIF tests with R. rickettsii and R. slovaca antigens and four-fold decrease after treatment.

[Ticks infected with bacteria pathogenic to humans in municipal parks in Warsaw]. / Kleszcze zakazone bakteriami chorobotwórczymi dla czlowieka na terenach parków miejskich warszawy.

The aim of the study was to detect bacteria pathogenic to humans, such as Borrelia burgdorferi sensu lato, Rickettsia spp, and Coxiella burnetii in ticks found in municipal parks in Warsaw, including The Royal Lazienki Park, Mokotów Fields, Józef Sowinski Park, Ujazdów Park and Fort Bema Park. To detect microorganisms PCR technique was used. It was shown that 6.1% of the ticks in Warsaw's parks are infected with B. burgdorferi sensu lato, and 2.9% with Rickettsia spp. Analysis of sequencing results revealed in examined ticks the presence of B. burgdorferi sensu stricto and R. helvetica.

Interactions between Borrelia burgdorferi and mouse fibroblasts.

Borrelia burgdorferi spirochetes are an infectious agent of Lyme borreliosis. The aim of our studies was to investigate the fate of engulfed B. burgdorferi cells in L-929 mouse fibroblasts and to observe development of intracellular infection in vitro after 2 and 48 h. Electron microscopic studies reveal consecutive stages of B. burgdorferi spirochetes penetration to mouse fibroblasts in vitro. It has been observed, as a first step attachment and engulfment of spirochetes followed by formation of vacuoles. After 48 hours of infection, vacuoles of fibroblastic cells have been seen full of B. burgdorferi bacteria and latter they have been released from infected cells to extracellular space. It can be the evidence that B. burgdorferi multiply intracellulary.

Rickettsia spp. in ticks, Poland.

Ticks are recognized as the main vectors and reservoirs of spotted fever group rickettsiae. We searched for the most prevalent Rickettsia spp. in Poland and found R. slovaca and R. helvetica bacteria in ticks in southern and central Poland; R. raoulti was found in ticks in all parts of Poland.