Blood parasites of water frogs (Pelophylax esculentus complex) from the Danube Delta, Romania.

Water frogs of the genus Pelophylax host a variety of parasites, from protozoa to helminths. Among the blood parasites, representatives of Apicomplexa, Trypanosoma and Nematoda show the highest prevalence. In this study, we focused on blood parasites of water frogs living in the Danube Delta, Romania. In total, 74 individuals of P. ridibundus and eight individuals of P. esculentus from six localities were examined. Blood parasites were detected microscopically and using a molecular marker (18S rDNA). 89.77% of frogs from all investigated localities were found to be infected with at least one parasitic group, specifically with haemogregarines (84.09%), nematodes (1.14%), and trypanosomes (63.64%). The parasitemia of haemogregarines and trypanosomes differed significantly among the studied locations. There was no statistically significant difference in parasitemia between male and female hosts. However, adults were found to have a significantly higher parasitemia in comparison with subadults infected with haemogregarines. Correlation between parasitemia and the body length of frogs infected with haemogregarines was also significant (r = 0.226). By comparing the 18S rDNA sequences with the corresponding GenBank sequences, Hepatozoon species identified in water frogs showed a close similarity (98.1-99.8%) to Hepatozoon magna. Trypanosomes showed the highest sequence similarity to Trypanosoma sp. isolate R10 clone L2-3, Trypanosoma ranarum, and Trypanosoma cobitis.

Effects of Electromagnetic Radiation on Neuropeptide Transcript Levels in the Synganglion of Ixodes ricinus.

Anthropogenic electromagnetic radiation is an important environmental factor affecting the functionality of biological systems. Sensitivity to various frequencies of electromagnetic radiation has been detected in ixodid ticks in the past. However, the physiological aspects of radiation effects have not yet been studied in ticks. In the presented experiment, 360 Ixodes ricinus ticks, 180 males and 180 females, were divided into 16 irradiated and 8 control groups. The irradiated groups were exposed to two different intensities of electromagnetic radiation with a frequency of 900 MHz at different lengths of exposure time. RT-PCR was utilized to determine the changes in mRNA levels in tick synganglia after irradiation. Four randomly selected neuropeptide genes were tested-allatotropin (at), FGLa-related allatostatins (fgla/ast), kinin, and arginine-vasopressin-like peptide (avpl). A significant decrease in transcript levels in all female groups exposed to higher intensity radiofrequency radiation for 1 to 3 h was found. After one hour of radiofrequency exposure, a significant downregulation in allatotropin expression in males was detected. A consistent downregulation of the at gene was detected in males irradiated with at a higher intensity. Unfortunately, the specific functions of the studied neuropeptides in ticks are not known yet, so a more comprehensive study is necessary to describe the effects of EMF on observed neuropeptides. This study represents the first report on the effects of the abiotic environment on tick neurophysiology.

Interspecific differences in the behavioral response of ticks exposed to radiofrequency electromagnetic radiation.

Artificial electromagnetic radiation is a new environmental factor that affects animals. Experiments with the effect of radio frequency electromagnetic radiation were focused on both vertebrates and invertebrates. Ticks showed a significant affinity to radiation. Our study is a continuation of this research and its aim was to monitor the effect of radiation on the behavior of four tick species: Ixodes ricinus, Dermacentor reticulatus, Dermacentor marginatus and Haemaphysalis inermis. In total 1,200 ticks, 300 of each species, were tested in modules allowing the choice of an exposed or shielded area. During the test, the ticks were exposed to electro-magnetic radiation of 900 MHz for 24 h. The position of the individuals was recorded and we evaluated the obtained data statistically. We observed a significant preference to the exposed area in both sexes of I. ricinus. Males of D. reticulatus and D. marginatus also showed an affinity to radiation, but not females of both species, nor females and males of H. inermis. The results of the study support the assumption that ticks perceive the electromagnetic field and the observed differences in their response have the potential to help understand the mechanism of perception.

The Behavior of Rickettsia-Positive Dermacentor reticulatus Ticks under Laboratory Conditions.

Considering that tick-borne diseases are on the rise, a full understanding of how pathogen-tick-host interactions can lead to vector behavioral modifications is of high value. Successful transmission of pathogens to their hosts depends on vector mobility and their ability to quest for prey and attack hosts. In our research, the behavior of Dermacentor reticulatus ticks (n = 88) that were positive or negative for Rickettsia spp. (prevalence-36.36%) were analyzed using novel behavioral apparatuses. Tick locomotion and their preference for light or darkness were studied utilizing a multirod arena (MA) and a light/dark test (LDT) optimized for ticks. Behavioral tasks were evaluated using a Smart video-tracking system (Panlab, Spain). The majority of ticks (p < 0.0001) explored in the center of MA during the first 15 min. Despite that, most of them (p < 0.01) chose the periphery for questing or resting postures. They also preferred the elevated parts of the rods (p < 0.001) for this behavior. Ticks significantly (p < 0.0001) preferred the light part of the LDT. Rickettsia-infected ticks had higher locomotor activity, manifesting as longer trajectories (p = 0.0221). Our results revealed the possible impact of pathogens on some behavioral patterns of ticks as their vectors, which may significantly increase the probability of pathogen transmission.

Behavioral Repertoire on a Vertical Rod-An Ethogram in Dermacentor reticulatus Ticks.

Ticks are important vectors of pathogens that endanger humans and animals. Study of their behavior under laboratory conditions is important for both predicting their behavior in natural conditions and understanding their involvement in transmission cycles of pathogens, which may lead to effective prevention of tick-borne disease transmission or establishment of effective preventive measures. The aim of our study was to describe the behavior of D. reticulatus ticks using laboratory assay. We focused on the description of individual behavioral units during their vertical movement. The assay consisted of glass beakers filled with sand and an embedded glass rod. We observed 10 different behavioral units, 4 of which have not yet been described: body posturing called "jogger", leg grooming, and body or leg jerking. The most frequent tick behavior observed was an upwards positioning of the two front legs while the body remained motionless (88.9%). Other common observations were both horizontal (63%) and vertical (58.0%) body posturing with all legs lowered, followed by questing behavior (51.9%). Ticks spent the most time questing (75.2%), crawling (54.7%), and grooming legs on the right side (23%). We did not observe any differences between males and females.

Prevalence of relapsing fever spirochete Borrelia miyamotoi in Ixodes ricinus ticks from eastern Slovakia.

Borrelia miyamotoi, recurrent fever borrelia, pathogenic to humans, has been found recently in Ixodes ricinus ticks. There is still little information about its circulation in nature and potential local impact on human health as well as the occurrence in natural localities. In our study, a total of 1609 ticks (745 nymphs, 358 females and 506 males) collected from different localities in eastern Slovakia were analysed using a polymerase chain reaction targeting a gene encoding glycerophosphodiester phosphodiesterase (glpQ), which is specific to the Borrelia species in the relapsing fever group. B. miyamotoi was found in 3 nymphs, 6 females and 9 males of quested I. ricinus ticks. Although no human case of infection with B. miyamotoi has been reported in Slovakia yet, its presence needs to be studied to obtain knowledge regarding the prevalence and the distribution of this human pathogen in questing ticks and to increase medical healthcare awareness.

Morphometric analysis - effect of the radiofrequency interface of electromagnetic field on the size of hatched Dermacentor reticulatus larvae.

INTRODUCTION AND OBJECTIVE: Electromagnetic radiation interactions with living systems have been one of determining factors in biological evolution. This study investigates the effect of 900 MHz radiofrequency (RF) electromagnetics field (EMF) exposure of eggs on the development of Dermacentor reticulatus larvae. The basic objective was to determine whether the 900 MHz RF-EMF has the potential to influence the size of the body of the hatched larvae of D. reticulatus ticks. MATERIAL AND METHODS: To this aim, eggs from 3 fully engorged females of D. reticulatus were included in the test procedure. Altogether four groups of eggs were designated which included eggs from each female. We used RF-EMF frequency of 900 MHz. Eggs were exposed to EMF for different time periods (30, 60 and 90 minutes) in dark, electromagnetically shielded anechoic chamber. After the irradiation eggs were allowed to hatch in climatic chamber. Randomly selected 200 larval individuals were measured to get basic morphological records. Four body traits including the total body length (TBL), length of gnathosoma with scutum (GSL), the total body width (TBW), and the width of basis capituli (BCW) were measured. RESULTS: The D. reticulatus larvae hatched from eggs exposed for 60 minutes, had demonstrably larger dimensions of all measured body traits not only as a control unexposed group but also as other experimental groups. CONCLUSIONS: The study shows, particularly, that artificial EMF that is used in smartphone technology impacts seriously D. reticulatus larvae development.

Enlisting the Ixodes scapularis Embryonic ISE6 Cell Line to Investigate the Neuronal Basis of Tick-Pathogen Interactions.

Neuropeptides are small signaling molecules expressed in the tick central nervous system, i.e., the synganglion. The neuronal-like Ixodes scapularis embryonic cell line, ISE6, is an effective tool frequently used for examining tick-pathogen interactions. We detected 37 neuropeptide transcripts in the I. scapularis ISE6 cell line using in silico methods, and six of these neuropeptide genes were used for experimental validation. Among these six neuropeptide genes, the tachykinin-related peptide (TRP) of ISE6 cells varied in transcript expression depending on the infection strain of the tick-borne pathogen, Anaplasma phagocytophilum. The immunocytochemistry of TRP revealed cytoplasmic expression in a prominent ISE6 cell subpopulation. The presence of TRP was also confirmed in A. phagocytophilum-infected ISE6 cells. The in situ hybridization and immunohistochemistry of TRP of I. scapularis synganglion revealed expression in distinct neuronal cells. In addition, TRP immunoreaction was detected in axons exiting the synganglion via peripheral nerves as well as in hemal nerve-associated lateral segmental organs. The characterization of a complete Ixodes neuropeptidome in ISE6 cells may serve as an effective in vitro tool to study how tick-borne pathogens interact with synganglion components that are vital to tick physiology. Therefore, our current study is a potential stepping stone for in vivo experiments to further examine the neuronal basis of tick-pathogen interactions.