High Prevalence of Borrelia burgdorferi Antibodies in Jaworzno, Poland: A Retrospective Study Revealing Endemic Lyme Borreliosis.

BACKGROUND This retrospective study of 704 adult residents of Jaworzno, Poland, aimed to evaluate medical personnel awareness of episodes of Lyme borreliosis and serum antibody levels for Borrelia burgdorferi sensu lato. MATERIAL AND METHODS This study included 704 residents of Jaworzno, Poland, who had no more than 12 months between tick bite and screening. The study consisted of a self-designed questionnaire survey and an analysis of IgG and IgM antibodies against B. burgdorferi sensu lato using an enzyme-linked assay (ELISA) and Western blot analysis, when necessary, to confirm the results. RESULTS A total of 558 residents (79.3%) confirmed having contact with a tick, 84 (11.9%) responded that they did not remember having contact with a tick, and 62 (8.8%) denied having contact with a tick. Regarding IgG, the ELISA showed 183 (25.99%) positive, 440 (62.5%) negative, and 81 (11.5%) equivocal results. Regarding IgM, the ELISA showed 180 (25.57%) positive, 435 (61.79%) negative, and 89 (12.64%) equivocal results. Positive and equivocal results for the IgG and IgM classes using the ELISA test were confirmed in 36 cases (13.64%) for IgG and in 53 cases (19.70%) for IgM using Western blot analysis. CONCLUSIONS The ELISA method obtained similar values for positive, negative, and equivocal results in the serological test. This was reflected in the survey conducted on residents who reported a tick bite and later received a positive result in the ELISA test as well as an approximate time between the bite and removal of the tick.

The Identification of Potential Immunogenic Antigens in Particular Active Developmental Stages of the Rice Weevil (Sitophilus oryzae).

BACKGROUND: The rice weevil (Sitophilus oryzae) originates from subtropical and tropical areas of Asia and Africa, but it also appears on other continents, mostly as a result of trade in rice. It may occur in grain fields as well as in storage facilities, and cause allergenic reactions. The aim of this study was to identify the potential antigens in all developmental stages of S. oryzae, which may cause an allergic response in humans. METHODS: Sera of 30 patients were tested for the presence of IgE antibodies to antigens from three life stages of the rice weevil. To identify protein fractions containing potential allergens, proteins collected from larvae, pupae, and adults separated by sex of S. oryzae were fractionated by SDS-PAGE. Then, they were probed with anti-human, anti-IgE monoclonal antibodies, fractionated by SDS-PAGE and detected by Western blotting. RESULTS: In total, 26 protein fractions of males and 22 fractions of other life stages of S. oryzae (larvae, pupae, and females) positively reacted with the examined sera. CONCLUSIONS: The conducted study showed that S. oryzae may be a source of many antigens which may cause the potential allergic reactions in humans.

Back to Locality? Demand Potential Analysis for Short Food Supply Chains.

The main purpose of the article is an attempt to estimate the demand for products traded within short food supply chains in Poland. The survey was conducted in autumn 2021 in the Kamienna Góra county, where the first business incubator in Poland, addressed to farmers and food producers, initiated and supported by the local government, is located. The Computer-Assisted Web Interview (CAWI) method constituted the basis for the process of collecting research material. The channel for reaching respondents was the LIBRUS application and local social media. Responses were mainly given by women, people with incomes in the range of PLN 1000-3000 per person, those aged 30-50, and those with a university education. The research results showed a high level of potential demand for local agri-food products, which should encourage farmers to switch from long to short supply chain models. The persisting low awareness regarding the existence of alternative distribution networks for local products, which primarily requires increased activities in the field of territorial marketing that promote local agri-food products among the residents of municipalities constitutes, from the perspective of consumers, a barrier to the development of short food supply chains.

The First Records of Canine Babesiosis in Dogs from Dermacentor reticulatus-Free Zone in Poland.

Tick-borne microorganisms belong to important etiological agents of many infectious diseases affecting humans and animals. Among them, there are haemoprotozoans of the Babesia genus, which infect erythrocytes of a host and may cause many clinical symptoms. Canine babesiosis is an emerging tick-borne disease in Southern and Central Europe. In this study, we report two cases of symptomatic canine babesiosis caused by Babesia canis in domestic dogs from the Silesian Voivodeship, Poland, as well as the presence of Dermacentor reticulatus ticks detected on one of the Babesia-infected dogs (D. reticulatus-free zone). The molecular analysis confirmed the presence of Babesia canis in the dogs' blood, and the sequencing analysis showed that the obtained sequence is 100% identical to the sequence of Babesia canis isolate 3469 (sequence ID: KX712122.1). Our findings should raise awareness of B. canis infection among dog owners and veterinarians in the region where B. canis was not previously reported in residential, non-traveling dogs, as well as ensuring that adequate diagnostic methods are available.

Abundance of domestic mites in dwellings of children and adolescents with asthma in relation to environmental factors and allergy symptoms.

Exposure to house dust allergens, mainly from domestic mites, is an important cause of allergic reactions in sensitized asthmatic patients. A total of 63 dust samples were collected from 16 flats in Bytom (south Poland); in each flat a person (age 4-17 years) suffering from bronchial asthma lived with his/her family. Mite density was calculated as the number of specimens per g of dust. The results were compared with household features and the data were statistically analyzed. In total 566 mite specimens were isolated, including 526 members of the family Pyroglyphidae (93%). The dominant species were Dermatophagoides pteronyssinus (60% of the total count) and Dermatophagoides farinae (32%). Pyroglyphids were found in all mite positive samples (68%) of which 35% also contained non-pyroglyphids, including glycyphagids, cheyletids and gamasids. The results suggest associations between the density of some mite taxa (per g of dust) and the following indoor environmental factors: presence of pets, number of inhabitants, coal-stoves as a type of heating, cleaning frequency, higher relative humidity, presence of flowers and PVC windows. The severity of asthma seems to be associated with the numbers of D. farinae, total domestic mites and live mites per g of dust.

The potential risk of exposure to Borrelia garinii, Anaplasma phagocytophilum and Babesia microti in the Wolinski National Park (north-western Poland).

Ixodes ricinus (Acari: Ixodida) is the main vector in Europe of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. Wolinski National Park (WNP) is situated by the Baltic Sea and is frequently visited by tourists. The aim of the study was to determine the potential risk of exposure to tick borne infection with B. burgdorferi s.l., A. phagocytophilum and B. microti on the areas of WNP. In total, 394 I. ricinus were tested. The pathogens in ticks were detected by PCR, nested PCR, RFLP and sequencing. Altogether, pathogens were detected in 12.69% of the studied ticks. B. burgdorferi s.l., was shown in 0.25% of the studied I. ricinus, while A. phagocytophilum and B. microti were detected in 1.01% and 10.65% of studied ticks, respectively. Co-infection by A. phagocytophilum and B. microti was shown in only one I. ricinus nymph. Analysis of B. burgdorferi s.l., genospecies showed that 0.25% of the studied ticks were infected with Borrelia garinii. The obtained results show the potentially high human risk of exposure to tick-borne infection with B. microti, and the low potential risk of infection with B. garinii and A. phagocytophilum on the studied areas of WNP.

Comparison of the domestic mites abundance in dwellings on selected urban and rural areas of the Zawiercie district (south-west Poland).

Mites are found in all types of environments, inhabiting also the immediate human environments, including dust from sleeping accommodations, upholstered furniture or carpeted floors. It is commonly known that house dust mites are sources of potent inhalant allergens. Dermatophagoides pteronyssinus and Dermatophagoides farinae belong to the most common species in the temperate climate zone. Samples were collected by using a portable vacuum cleaner, into trap-filters installed onto the cap for dusting. Then, mites were isolated by a flotation method with saturated NaCl solution. The Petri dishes were screening under a stereoscopic microscope for presence of mites, then isolated mites were mounted on microscopic slides in Hoyer's medium. Mites were identified to species under the light microscope using phase contrast optics. A total of 724 mites were isolated from 46 of the examined samples, including 666 (91.9%) members of the family Pyroglyphidae. Among them D. pteronyssinus was predominat (62.8% of the total count, with average number 211.22 specimens per 1 gram of dust), followed by D. farinae (24.03%, averagely 150.07 specimens per 1 gram of dust) and Euroglyphus maynei (4.42%, 118.11 specimens per 1 gram of dust). Higher density of house dust mites was associated with the following factors: higher age of building, higher number of roommates, higher washing frequency per week, presence of carpeted floor in bedrooms, wooden floors in kitchens, closed kitchen, uphoplstered furniture, absence of pets and unemployed housewives.

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.

Morphology Reveals the Unexpected Cryptic Diversity in Ceratophyllus gallinae (Schrank, 1803) Infested Cyanistes caeruleus Linnaeus, 1758 Nest Boxes.

PURPOSE: The main aim of our study was to examine morphological differentiation between and within sex of hen fleas-Ceratophyllus gallinae (Schrank, 1803) population collected from Eurasian blue tit (Cyanistes caeruleus Linnaeus, 1758), inhabiting nest boxes and to determine the morphological parameters differentiating this population. METHODS: A total of 296 fleas were collected (148 females and 148 males), determined to species and sex, then the following characters were measured in each of the examined fleas: body length, body width, length of head, width of head, length of comb, height of comb, length of tarsus, length of thorax and length of abdomen. RESULTS: The comparison of body size showed the presence of two groups among female and male life forms of the hen flea, which mostly differed in length of abdomen, whereas the length of head and tarsus III were less variable. CONCLUSION: Till now, the only certain information is the presence of two adult life forms of C. gallinae. The genesis of their creation is still unknown and we are not able to identify the mechanism responsible for the morphological differentiation of fleas collected from the same host. In order to find answer to this question, future research in the field of molecular taxonomy is required.

The Discovery of Zoonotic Protozoans in Fleas Parasitizing on Pets as a Potential Infection Threat.

PURPOSE: Fleas are insects with a high medical and veterinary importance. They may participate in spreading of many pathogenic agents, but still there is limited information about their possible reservoir or vector role for protozoans. The main aim of this study was an attempt of detection zoonotic pathogens, such as Babesia microti and Toxoplasma gondii in fleas Ctenocephalides felis felis and Ctenocephalides canis. METHODS: In 2013-2017, 155 fleas were captured from domestic dogs and cats in veterinary clinics, animal shelters and pet grooming salons in Upper Silesia Region in Poland. Then, the DNA was extracted from each Ctenocephalides flea by using the ammonia method. Samples were screened for the presence of B. microti and T. gondii using PCR and nested PCR methods. RESULTS: B. microti was reported in 6.6% of C. felis felis and 9.1% of C. canis, whereas the prevalence of coinfection with B. microti and T. gondii was 1.9% in cat fleas and 2.3% in dog fleas. CONCLUSION: This study shows the first cases of B. microti occurrence and B. microti and T. gondii coinfection in Ctenocephalides fleas. The estimation of prevalence of examined protozoans may be useful considering the possibility of infection among companion animals, as well as during presentation of the potential risk of infection in humans. In order to clarify the role of C. felis felis and C. canis in transmission of B. microti and T. gondii, the another studies with in vitro cultures and laboratory animals are needed.

What Are the Costs of Diagnostics and Treatment of Lyme Borreliosis in Poland?

Lyme borreliosis (LB) is a growing epidemiological threat in many areas of the world, including North America and Europe. Due to the lack of effective protection against this disease, it seems important to ensure a timely diagnosis for effective treatment and the prevention of serious health consequences. The aim of this study was to assess the costs of diagnosis and treatment of Lyme disease in Poland. The costs incurred for the medical payer were analyzed. The cost of hospitalization due to LB for one patient in 2018 was estimated to be ~582.39 EUR, which constituted 53.10% of the average monthly salary of that year. In the analyzed period (2008-2018), the number of people treated by medical services due to Lyme disease increased, both in hospitalization and ambulatory specialist care. Although, the costs of hospitalization were the highest of the two, we noticed a change ratio between hospitalization and ambulatory specialist care in favor of the latter.

The molecular detection of Anaplasma phagocytophilum and Rickettsia spp. in cat and dog fleas collected from companion animals.

Companion animals can be infested by various species of parasitic insects. Cat flea Ctenocephalides felis (C. felis felis) (Bouché, 1835) and dog flea Ctenocephalides canis (Curtis, 1826) belong to multihost external parasites of mammals, which most frequently occur on domestic cats Felis catus Linnaeus and dogs Canis familiaris Linnaeus. The main aim of this study was to investigate the presence of pathogens, such as Anaplasma phagocytophilum (syn. Ehrlichia phagocytophila) and Rickettsia spp., in adult C. felis and C. canis fleas. Flea sampling has been realised from January 2013 to April 2017 in veterinary clinics, animal shelters and pet grooming salons. Fleas were collected from domestic cats and dogs, directly from the pet skin or hair. Then, the DNA was isolated from a single flea by using the alkaline hydrolysis and samples were screened for the presence of pathogens using PCR method. Anaplasma phagocytophilum has occurred in 29% of examined C. felis and 16% of C. canis individuals. In turn, the prevalence of Rickettsia spp. in cat fleas population was only 3%, and the dog fleas 7%. The present study showed the presence of pathogenic agents in cat and dog fleas, which indicates the potential role of these insects in circulation of A. phagocytophilum and Rickettsia spp. in the natural habitat. Furthermore, exposition to these flea species, whose hosts are domestic cats and dogs, can pose a potential risk of infection for humans.

Occurrence of Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti in Ixodes ricinus ticks collected from selected areas of Opolskie Province in south-west Poland.

INTRODUCTION: Ticks (Acari: Ixodida) are vectors and/or reservoirs of many pathogens, i.e. Borrelia burgdorferi sensu lato, Anaplasma phagocytophilum and Babesia microti. These pathogens are ethiological agents of such diseases as Lyme borreliosis, human granulocytic anaplasmosis and human babesiosis. OBJECTIVE: The aim of the study was to evaluate the role of the Ixodes ricinus in the transmission of Borrelia burgdorferi sensu lato, Borrelia afzelii, Borrelia garinii, Borrelia burgdorferi sensu stricto, Anaplasma phagocytophilum and Babesia microti in Opolskie Province in Poland. MATERIAL AND METHODS: DNA from 222 ticks was isolated by the ammonia method. The pair of primers specific to the flagelline gene was used to detect of B. burgdorferi s. l. To detect of genospecies of this spirochete, three pairs of internal primers were used. In turn, two pairs of primers specific to the 16S rDNA gene and the 18S rRNA were used, respectively, for the detection of A. phagocytophilum and B. microti. Borrelia burgdorferi s. l., A. phagocytophilum, and B. microti were detected in 4.5%, 2.7% and 5.4% of examined ticks, respectively. RESULTS AND CONCLUSIONS: Of the ten ticks infected with B. burgdorferi s. l., B. afzelii was found in seven, undefinied genospecies in two, and mixed infection with B. afzelii and B. burgdorferi s. s. in one. The study demonstrated the potential risk of exposure of humans and animals to infections of B. burgdorferi s. l., A. phagocytophilum and B. microti in the examined area of Poland.

Risk of exposure of a selected rural population in South Poland to allergenic mites. Part II: acarofauna of farm buildings.

Exposure to mite allergens, especially from storage and dust mites, has been recognized as a risk factor for sensitization and allergy symptoms that could develop into asthma. The aim of this study was to investigate the occurrence of mites in debris and litter from selected farm buildings of the Malopolskie province, South Poland, with particular reference to allergenic and/or parasitic species as a potential risk factor of diseases among farmers. Sixty samples of various materials (organic dust, litter, debris and residues) from farm buildings (cowsheds, barns, chaff-cutter buildings, pigsties and poultry houses) were subjected to acarological examination. The samples were collected in Lachowice and Kurów (Suski district, Malopolskie). A total of 16,719 mites were isolated including specimens from the cohort Astigmatina (27 species) which comprised species considered as allergenic (e.g., Acarus siro complex, Tyrophagus putrescentiae, Lepidoglyphus destructor, Glycyphagus domesticus, Chortoglyphus arcuatus and Gymnoglyphus longior). Species of the families Acaridae (A. siro, A. farris and A. immobilis), Glycyphagidae (G. domesticus, L. destructor and L. michaeli) and Chortoglyphidae (C. arcuatus) have been found as numerically dominant among astigmatid mites. The majority of mites were found in cowsheds (approx. 32%) and in pigsties (25.9%). The remaining mites were found in barns (19.6%), chaff-cutter buildings (13.9%) and poultry houses (8.8%). The results suggest that the allergenic mites may constitute an occupational hazard for agricultural workers in all farming environments examined.

Risk of exposure of a selected rural population in South Poland to allergenic mites. Part I: indoor acarofauna of one-family houses.

The aim of this study was to investigate the occurrence of mites in dust samples from houses in agricultural areas of South Poland, with particular reference to allergenic and parasitic species as a potential risk factor of diseases among people. A total of 250 dust samples from 50 single-family houses situated in Stryszawa and vicinity (Malopolskie province) were examined for the presence of domestic mites. Dust was taken from beds, floors in bedrooms, upholstery furniture, floors in family rooms and from floors in kitchens. Mites were found in 74.8% of samples collected. A total of 5340 mite specimens were isolated, including 2771 members of the family Pyroglyphidae (51.9%). Dominants were Dermatophagoides pteronyssinus (36.4% of all mites) and Gohieria fusca (25.1%), followed by Chortoglyphus arcuatus (18.3%) and D. farinae (15.1%). Dermatophagoides pteronyssinus was also the most frequent species (53.2% of the total count of samples examined), followed by G. fusca (42.4%), D. farinae (37.2%) and C. arcuatus (36.4%). Lepidoglyphus destructor was found more frequently than Glycyphagus domesticus in the examined samples. Density of D. pteronyssinus was associated with beds, presence of pets, coal stoves used for heating, lower number of rooms, higher cooking frequency, higher washing frequency, working housewife, open kitchen, wooden floors in kitchens, lower cleaning frequency, type of upholstery furniture in living rooms (arm chairs), lower humidity and higher temperature.

[Significance of kallikrein-kinin system in central nervous system diseases].

Kallikreins cleave kininogens to release kinins. Kinins exert their biological effect by activating constitutive bradykinin receptor-2 (BR2) and inducible by inflammatory cytokines bradykinin receptor-1 (BR1). Studies in animal models and some clinical observations indicate tat the activation of kallikrein - kinin system may have relevance to central nervous system (CNS) diseases, including multiple sclerosis, Alzheimer's disease, epilepsy as well as cerebral ischemia and neoplasmatic tumors. The actions of kinins include vasodilatation and increased vascular permeability may contribute to blood-brain barrier disruption. Kinins evoke pain, and stimulate of endothelial cells, white blood cells, astrocytes and microgia cells to release of prostanoids, cytokines, free radicals, nitric oxide. Kinins stimulate angiogenesis and proliferation of tumor cells. These events lead to neural tissue damage and long lasting disturbances in blood-brain barrier function. In animal models the overexpression of genes and proteins of tissue kallikrens, kininogen as well as RB1 and RB2 has been observed. Kinin receptors antagonists, especially B1R blockade decreased morphological and biochemical features of CNS inflammation. On the other hand in brain tumor models RB1 and RB2 activation has been shown to mediate reversible blood-brain barrier permeability to enhance anti-cancer drug delivery, which may have therapeutic potential.

Sensitization to the storage mites Lepidoglyphus destructor and Tyrophagus putrescentiae (Acari, Sarcoptiformes, Astigmatina) in a suburban population in Southern Poland

Mite infestation of stored products is a serious threat to food safety and public health. These stored product mites are not only serious pests of stored food but also cause allergies in humans. Thirty serum samples from patients living in suburban areas of Upper Silesia (South Poland) were tested for sensitization to two species of storage mites: Lepidoglyphus destructor [LD] and Tyrophagus putrescentiae [TP]. Patient antibodies against particular antigens were identified using anti-human anti-IgE monoclonal antibodies. Fifteen protein fractions from LD gave positive reactions with IgE antibodies and 18 from TP. Seven of the 30 samples showed positive reactions to a protein fraction measuring about 29 kDa from LD and six reacted with a fraction measuring about 25 kDa from TP. These findings may imply the existence of many protein fractions with allergenic properties besides the characterized allergens in the two tested species.

Expression Profiles of Toll-Like Receptors in the Differentiation of an Infection with Borrelia burgdorferi Sensu Lato Spirochetes.

The similarity of Lyme borreliosis to other diseases and its complex pathogenesis present diagnostic and therapeutic difficulties. The changes that occur at the cellular and molecular levels after a Borrelia sp. infection still remain poorly understood. Therefore, the present study focused on the expression of TLR and TLR-signaling genes in human dermal fibroblasts in the differentiation of an infection with Borrelia burgdorferi sensu lato spirochetes. Normal human dermal fibroblasts were cultured with the spirochetes of Borrelia burgdorferi sensu stricto, Borrelia afzelii and Borrelia garinii. Total RNA was extracted from the cells using TRIzol reagent. The analysis of the expression profiles of TLRs and TLR-related genes was performed using commercially available oligonucleotide microarrays of HG-U133A. The GeneSpring 12.0 platform and significance analysis of microarrays were used for the statistical analysis of microarray data. The analyses using the oligonucleotide microarray and QRT-PCR techniques permitted to identify the genes encoding TLR4 and TLR6 as specific for infection with B. afzelii and B. burgdorferi sensu stricto. In turn, TLR3 was only characteristic for an infection with B. burgdorferi sensu stricto. There were no changes in the TLR gene expression after infection with B. garinii. Our findings confirm that Borrelia has a major effect on fibroblast gene expression. Further characterization of changes in gene expression may lead to valuable insights into the role of the toll-like receptor in the pathogenesis of Lyme disease and may provide guidelines for the development of diagnostic markers for an infection with a particular Borrelia genospecies. Moreover, this will help to identify better treatment strategies for Lyme disease.

The risk of exposure to parasitic mites and insects occurring on pets in Southern Poland

Companion animals, including pets, can be infested by many species of parasitic mites and insects, which can pose a threat to the health of both animals and humans. The aim of this study is to evaluate the risk of exposure of companion animals to various species of external parasites which can be transferred to humans and cause many skin diseases. External parasites were collected in 2012­2014 from the patients of veterinary clinics in the provinces of Silesia and Malopolska (Southern Poland). Parasitic mites and insects were collected using preparation needles. Parasites were classified to species and life stage under a Stemi 2000-C stereomicroscope (Zeiss) or an ECLIPSE E-200 optical microscope (Nikon). They were then analysed by an Optika Vision Pro system (Nikon). In total, 77 samples were taken: 62 from the Malopolska and 15 from Silesia. A total of 999 specimens of parasitic mites and 225 specimens of parasitic insects were isolated from the samples. The dominant mite species was Otodectes cynotis var. cati (Astigmatina, Psoroptidae) with 855 isolated specimens (85.6% of the total number of isolated mites). Polyplax spinulosa was the predominant parasitic insect species: 209 specimens were identified, constituting 92.9% of all examined insects. Our findings indicate that companion animals and their owners have a high risk of exposure to ectoparasites.