The Analysis of Selected miRNAs and Target MDM2 Gene Expression in Oral Squamous Cell Carcinoma.

MiRNAs could play an important role in tumorigenesis and progression. The oncoprotein MDM2 (murine double minute 2) was identified as a negative regulator of the tumour suppressor p53. This study aims to analyse the expression of the MDM2 target miRNA candidates (miR-3613-3p, miR-371b-5p and miR-3658) and the MDM2 gene in oral squamous cell carcinoma tumour and margin samples and their association with the selected socio-demographic and clinicopathological characteristics. The study group consisted of 50 patients. The miRNAs and MDM2 gene expression levels were assessed by qPCR. The expression analysis of the miRNAs showed the expression of only one of them, i.e., miR-3613-3p. We found no statistically significant differences in the miR-3613-3p expression in tumour samples compared to the margin samples. When analysing the effect of smoking on miR-3613-3p expression, we demonstrated a statistically significant difference between smokers and non-smokers. In addition, we showed an association between the miR-3613-3p expression level and some clinical parameters in tumour samples (T, N and G). Our study demonstrates that miR-3613-3p overexpression is involved in the tumour progression of OSCC. This indicates that miR-3613-3p possesses potential prognostic values.

Lower dose of plant substance more effective in repelling Rhyzopertha dominica F . (Coleoptera, Bostrichidae) and Sitophilus granarius L. (Coleoptera, Dryophthoridae).

INTRODUCTION AND OBJECTIVE: The study compares the effect of two concentrations (0.1% and 1%) of plant material on repelling two dangerous stored pests - Rhyzopertha dominica F. and Sitophilus granarius L. Both species were treated with the essential oils of Foeniculum vulgare Mill., Carum carvi L., Mentha pulegium L., and Cananga odorata (Lam.) Hook.f. & Thomson, as well as the chemicals contained in these oils: anethole, L-carvone, pulegone and thymoquinone. MATERIAL AND METHODS: Laboratory tests were carried out at 29±1 °C with 60±5% relative humidity (RH). Repellence of insects were noted after 1, 2, 3, 4, 5, 24 and 48 h. RESULTS: A lower dose of essential oils and some of the tested compounds caused a stronger repellence in the specified species of beetles. In the case of R. dominica, the concentration of 0.1% pulegone and L-carvone had the greatest repellent effect. Differences in the repellence of R. dominica were found between the concentrations of 0.1%-1% among all the tested substances in each time interval. This difference ranged from 5.0%-52.92%. All substances used in the tests had a strong repellent effect on S. granarius (30%-100%). There were differences in the repellency of S. granarius between 0.1%-1% concentration in most of the tested substances in each time interval, but they were not as large as for R. dominica.. CONCLUSIONS: The most interesting results were obtained concerning the relationship: the lower tested substance concentration caused a much stronger repellent effect of R. dominica and S. granarius..

Review of ticks attacking domestic dogs and cats, and their epidemiological role in the transmission of tick-borne pathogens in Poland.

INTRODUCTION AND OBJECTIVE: Progressive climate change plays a major role in the expansion of tick populations, as well as the transmission of tick-borne disease pathogens (TBPs) to humans and animals throughout the world. Zoonoses are a growing environmental problem of great importance for public health. In Poland, domestic dogs and cats are mainly infested by Ixodes ricinus, Ixodes crenulatus, Ixodes hexagonus, Ixodes rugicollis and Dermacentor reticulatus ticks from the Ixodidae family. Other tick species, such as I. apronophorus and Haemaphysalis concinna, recorded sporadically on domestic dogs or cats, may expand their range in the future and successfully infest pets. Individual cases of infestations in Poland with foreign tick species, such as Rhipicephalus sanguineus, are known and may soon be recorded more frequently. OBJECTIVE: The aim of this review is to summarize the occurrence and identification of tick species and TBDs that are present or are likely to spread in Poland, which, given the medical and veterinary importance of ticks, may help in the development of public health strategies. REVIEW METHODS: A comprehensive literature review of publications and own research and data analysis was performed, obtained from reports and scientific descriptions on the epidemiology of tick-borne diseases. BRIEF DESCRIPTION OF THE STATE OF KNOWLEDGE: The ecology of ticks and hosts in urban and suburban environments is critical for quantifying the parameters necessary for an initial risk assessment, and identifying public health strategies to control and prevent TBDs. In the future, these species may expand their range of occurrence and their host, becoming common members of the Polish tick fauna. SUMMARY: Anaplasma, Babesia, Borrelia and Rickettsia spp. are the major TBPs in Poland, and their prevalence is usually higher in dogs than cats.

Exposure of domestic dogs and cats to ticks (Acari: Ixodida) and selected tick-borne diseases in urban and recreational areas in southern Poland.

The public health problem of tick-borne diseases has attracted much attention in recent years due to an increasing incidence in humans and animals. The aim of this study was to compare the risk of exposure to ticks and tick-borne infections in dogs and cats in recreational and urbanized areas in the Lesser Poland and Silesian Provinces. For molecular testing for the presence of the selected pathogens, 207 I. ricinus females collected from 119 dogs and 50 cats, and 2 I. hexagonus females collected from 2 domestic dogs, were examined. Overall, A. phagocytophilum was found in 3.7% of the I. ricinus specimens, B. microti in 27.1%, and B. burgdorferi s.l. in 0.9%. In urban areas of both provinces, A. phagocytophilum was found in 4.8% of the I. ricinus specimens, B. microti in 41.6% and B. burgdorferi s.l. in 3.9%. Pathogens were detected B. microti in both studied I. hexagonus specimens. These findings may indicate the important role that these animals play in the circulation of these pathogens in nature.

Fivefold higher abundance of ticks (Acari: Ixodida) on the European roe deer (Capreolus capreolus L.) forest than field ecotypes.

The European roe deer (Capreolus capreolus) is the most common deer species in Europe. The species can be a reservoir of some tick-borne diseases but it is primarily recognized for its contribution as an amplifier host. In Central Europe, two roe deer ecotypes are living in adjacent areas: field and forest. We investigated differences in tick load and species composition on these two ecotypes. We collected ticks from 160 (80 the forest ecotype and 80 the field ecotype) roe deer culled in Wielkopolska Region (West-Central Poland). The most common was Ixodes ricinus (n = 1610; 99%) followed by Ixodes hexagonus (n = 22; 1%). The dominant life stage of the ticks was female. Prevalence was higher for forest roe deer. Mean number of ticks found on the forest ecotype was almost fivefold higher than on the field ecotype (3.75 ± 0.83 vs. 0.77 ± 0.20 ticks). The mean probability of tick occurrence was threefold higher in the forest (0.915 ± 0.050) than the field ecotype (0.279 ± 0.125). The most infested body parts of roe deer from both ecotypes were the neck and the head.

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.

Ticks (Acari: Ixodida) on birds (Aves) migrating through the Polish Baltic coast.

Seasonal bird (Aves) migration between breeding and wintering areas, often located on different continents, can facilitate the spreading of tick species (Acari: Ixodida) and of tick-borne pathogens. The aim of the study was to analyse the occurrence of ticks dispersed by birds migrating along the Polish Baltic coast during spring and autumn migration. Field research was conducted at the bird ringing station in Wicie, located on the middle of the Polish Baltic coast, in 2011 and 2012 during spring and autumn migration. A total of 2657 birds from 45 species was examined. The most common species inspected were European robin (Erithacus rubecula) (63.3%), song thrush (Turdus philomelos) (5.13%), and goldcrest (Regulus regulus) (4.5%). Overall, 3129 ticks belonging to six species were collected: Ixodes ricinus (1650 larvae, 1390 nymphs and 1 male), Ixodes frontalis (20 larvae, 20 nymphs), Ixodes arboricola (35 larvae), Dermacentor reticulatus (1 larva), and Haemaphysalis punctata (1 nymph). Ten larvae and one nymph could only be identified to the genus level Ixodes. Ticks were located on various parts of the head: on the corner of the beak (75.0%), near the eyes (14.6%), on the chin (4.4%), near the ears (4.4%), on the neck (1.1%), and in the beak (0.5%). The overall tick prevalence was 40.5%. The highest prevalence was for bird species feeding on the ground, covering a medium distance to wintering grounds and migrating at night. Statistically significant differences between the number of ticks and the sex of the host species were detected in blackbirds: males carried more parasites than females, both, during spring and autumn migration. The fact that I. ricinus and other ticks parasitize birds migrating through Poland extends the possibility of the spread of tick-borne diseases.

Differential associations of Borrelia species with European badgers (Meles meles) and raccoon dogs (Nyctereutes procyonoides) in western Poland.

European badgers and raccoon dogs and their associated ticks and lice were assayed for the presence of Lyme borreliosis and relapsing fever-group spirochete DNA in western Poland. Analyses of blood, ear-biopsy and liver samples revealed that 25% of 28 raccoon dogs and 12% of 34 badgers were PCR positive for borreliae. Borrelia garinii was the dominant species in raccoon dogs (62.5%), followed by B. afzelii (25%) and B. valaisiana (12.5%). PCR-positive badgers were infected only with B. afzelii. A total of 351 attached ticks was recovered from 23 (82%) of the raccoon dogs and 13 (38%) of the badgers. Using a nested PCR targeting the ITS2 fragments of Ixodes DNA, four Ixodes species were identified: I. ricinus, I. canisuga, I. hexagonus, and one provisionally named I. cf. kaiseri. Ixodes canisuga and I. ricinus prevailed on both host species. The highest infection prevalence was detected in I. ricinus, followed by I. canisuga and I. cf. kaiseri. Borrelia garinii and B. afzelii accounted for 61.6% and 30.1% of the infections detected in all PCR-positive ticks, respectively. Four other Borrelia species (B. burgdorferi sensu stricto, B. valaisiana, B. lusitaniae and B. miyamotoi) were detected only in I. ricinus from raccoon dogs. Moreover, Borrelia DNA, mostly B. garinii, was detected in 57 (81.4%) of 70 Trichodectes melis lice derived from 12 badgers. The detection of B. afzelii in one-half of PCR-positive biopsies reconfirms previous associations of this species with mammalian hosts, whereas the high prevalence of B. garinii in feeding lice and I. ricinus ticks (including larvae) demonstrates that both carnivores serve as hosts for B. garinii. The lack of B. garinii DNA in the tissues of badgers versus its prevalence in raccoon-dog biopsies, however, incriminates only the latter carnivore as a potential reservoir host.

Detection of Borrelia burgdorferi sensu lato and spotted fever group rickettsiae in hard ticks (Acari, Ixodidae) parasitizing bats in Poland.

A total of 491 Ixodes vespertilionis and 8 Ixodes ricinus collected from bats and cave walls in southern Poland between 2010 and 2012 were examined by the polymerase chain reaction for tick-transmitted pathogens. PCR analysis for Borrelia burgdorferi s.l., Rickettsia spp., and Anaplasma phagocytophilum yielded negative results for all I. vespertilionis. DNA of Rickettsia helvetica was detected in three specimens of I. ricinus attached to Rhinolophus hipposideros or Myotis myotis, while Borrelia garinii was found in one tick parasitizing Myotis daubentonii. These pathogens were recorded for the first time in hard ticks that parasitized bats.

Prevalence of Borrelia burgdorferi sensu lato in Ixodes ricinus ticks collected from southern Poland.

In 2008-2011 ticks were collected from southern Poland. Out of 6336 individuals collected and identified as Ixodes ricinus, 768 (2 larvae, 84 nymphs, 417 females, 265 males) were included in molecular study. The aim of this study was to investigate the prevalence and types of genospecies of Borrelia burgdorferi sensu lato in ticks. The polymerase chain reaction (PCR) was applied to detect the presence of pathogens in ticks. Subsequently the amplified DNA was digested with TasI enzyme. The infection rate was 15% (116) of examined ticks. PCR-RFLP analysis allowed distinguishing three genospecies of B. burgdorferi s.l.: B. burgdorferi sensu stricto, B. afzelii, and B. garinii. RFLP analyses of 116 positive samples revealed 96 (83%) monoinfections and 13 (11%) coinfections, whereas unidentified genospecies were present in 7 (6%) of positive samples. In the case of monoinfections, B. burgdorferi s.s. was the predominant species of pathogen in infected ticks - 61.4%. Other genospecies: B. garinii and B. afzelii were detected in 22.9% and 15.6% of the samples, respectively. To sum up, 15 % of ticks were infected by B. burgdorferi s.l which increases the risk of human infections in the recreational areas of southern Poland. Furthermore, there is a need to increase public awareness and implement more preventive measures concerning Lyme disease.

A biological/medical review of alien tick species (Acari: Ixodida) accidentally transferred to Poland.

Alien tick species of the genera Ixodes, Amblyomma, Hyalomma and Rhipicephalus are sporadically transferred to the territories of the Central European countries. A biological-medical review of the following alien tick species transferred to Poland is presented: Ixodes eldaricus, Ixodes festai, Amblyomma sphenodonti, Amblyomma exornatumn, Amblyomma flavomaculatum, Amblyomma latum, Amblyomma nuttalli, Amblyomma quadricavum, Amblyomma transversale, Amblyomma varanense, Hyalomma aegyptium, Hyalomma marginatum, Rhipicephalus rossicus and Rhipicephalus sanguineus. The transfers of tick species out of the areas of natural distribution are divided into natural transfers (e.g., migration of ticks on hosts) and accidental transfers (e.g., resulting from the transport of livestock animals, trade in exotic animals, and transfers on animals during travel). It is important to monitor occurrence of unknown tick species on hosts in Poland.

The patterns of seasonal activity of Ixodes vespertilionis (Acari: Ixodidae) on Rhinolophus hipposideros in nursery colonies.

The aim of this study was to describe the dynamics of the long-legged bat tick Ixodes vespertilionis infestation on the lesser horseshoe bat Rhinolophus hipposideros in 2 nursery colonies roosting in attics. Out of a total of 810 lesser horseshoe bats examined, 217 (26.8%) were found to be infested with a total of 464 I. vespertilionis individuals. The developmental stage most frequently found was the larva, followed by the nymph, and the adult female. Bats were significantly more frequently infested with I. vespertilionis ticks in the period April to May than in other months. In these months, all tick developmental stages were observed. During summer and autumn, only immature developmental stages were recorded, whilst in September and October larvae predominated. Considerable differences in tick load between nursery colonies were observed. The length of seasonal presence on bats, prevalence, and infestation intensity of I. vespertilionis on lesser horseshoe bats were higher in the nursery colony situated in close vicinity of a cave than in the colony situated far from the caves. The results suggest that the pattern of seasonal infestation of ticks on bats roosting in nursery colonies coincides with the seasonal activity of Rh. hipposideros in the caves. The first case of mixed infestation of the lesser horseshoe bat with I. vespertilionis and I. ricinus were also recorded.

Ticks of Poland. Review of contemporary issues and latest research.

The paper presents current knowledge of ticks occurring in Poland, their medical importance, and a review of recent studies implemented in the Polish research centres on ticks and their significance in the epidemiology of transmissible diseases. In the Polish fauna there are 19 species of ticks (Ixodida) recognized as existing permanently in our country: Argas reflexus, Argas polonicus, Carios vespertilionis, Ixodes trianguliceps, Ixodes arboricola, Ixodes crenulatus, Ixodes hexagonus, Ixodes lividus, Ixodes rugicollis, Ixodes caledonicus, Ixodes frontalis, Ixodes simplex, Ixodes vespertilionis, Ixodes apronophorus, Ixodes persulcatus, Ixodes ricinus, Haemaphysalis punctata, Haemaphysalis concinna, Dermacentor reticulatus. Occasionally, alien species of ticks transferred to the territory of Poland are recorded: Amblyomma sphenodonti, Amblyomma exornatum, Amblyomma flavomaculatum, Amblyomma latum, Amblyomma nuttalli, Amblyomma quadricavum, Amblyomma transversale, Amblyomma varanensis, Amblyomma spp., Dermacentor marginatus, Hyalomma aegyptium, Hyalomma marginatum, Ixodes eldaricus, Ixodes festai, Rhipicephalus rossicus, Rhipicephalus sanguineus. The most common species of the highest medical and veterinary importance in Poland is invariably Ixodes ricinus. The review also sets out information on the risks of tickborne diseases in recreational areas of large cities in Poland, ticks as the cause of occupational diseases and dangerous species of ticks attacking people outside the Polish borders. Selected problems of the biology of ticks, the spread of alien species transferred on hosts and prevention of tick attacks have also been presented. The Polish studies on ticks are a valuable contribution to global research on the Ixodida.

Ixodes eldaricus Djaparidze, 1950 (Ixodidae) on migrating birds--reported first time in Poland.

During the ornithological "Operation Baltic" on the Hel Peninsula (the Baltic Sea coast in Poland) the first case of transfer to Poland of ticks of the species Ixodes eldaricus Djaparidze, 1950, on Prunella modularis (one female tick) and Erithacus rubecula (two males and one female tick). P. modularis and E. rubecula have not previously been recorded among the hosts of this tick species. Although the natural populations of I. eldaricus are very distant from Poland, it should be taken into account that this and other species of ticks may be transferred every year on migratory birds in the southern part of Central Europe and further north. Each case requires faunistic observation and epidemiological and morphological studies to exclude diagnostic confusion between very similar morphology in specimens of the genus Ixodes.