Agnieszka Jodełko

Tick-borne diseases (TBDs) are important infections affecting animal health, and some are zoonotic. Understanding the distribution of TBDs in wildlife is important to evaluate the role of wild animals in the maintenance and spread of tick-borne pathogens (TBPs) in natural ecosystems. This study investigates the occurrence of selected TBPs, including Anaplasma phagocytophilum and piroplasms (Babesia spp. and Theileria spp.), in three widespread wild cervids from Poland: red deer, fallow deer, and roe deer. Ninety spleen tissue samples (30 per species) were collected and analyzed by endpoint PCR and sequencing. DNA of A. phagocytophilum was detected in 54.6% of the samples, mainly in red deer (75.9%), followed by fallow deer (46.4%) and roe deer (41.4%). Piroplasm DNA was found in 30% of the samples, with the highest prevalence in red deer (47%), followed by fallow deer (27%) and roe deer (17%). Sequencing analysis showed the presence of Theileria capreoli and identified two distinct genotypes: “elaphi-CE1”, associated with red deer and fallow deer, and “capreoli-CE1”, associated with roe deer. The results update the epidemiology of TBPs in Poland, supporting the role of cervids as potential reservoirs of A. phagocytophilum and T. capreoli, confirming the need for surveillance in wildlife. Further studies are needed to investigate the distribution of T. capreoli genotypes across Europe.

Introduction: Different Chlamydia species affect cattle and contribute to economic losses. One of them, C. pecorum , is a globally endemic livestock pathogen. Despite its endemicity, prevalence data from Poland have so far been limited. The present study aimed to obtain insight into the chlamydiae prevalence in Polish dairy cattle. Material and Methods: A screening of chlamydial seroprevalence in dairy cattle was initially performed, followed by Chlamydiaceae - and species-specific real-time qPCR. Vaginal swabs (n = 239) and placenta samples (n = 2) from seropositive animals in 142 herds were collected to detect shedders. The study population consisted of cows (n = 2,780) from dairy herds (n = 1,153) located in all Polish voivodeships. Results: The true animal prevalence was determined to be 33.3%, while the true herd prevalence was 42.7%. Five groups of Polish voivodeships were identified using appropriate statistical tools, highlighting differences that may arise from various factors impacting the spread of chlamydial infections. The only detected chlamydia species was C. pecorum , the presence of which was confirmed in two herds. Conclusion: This study revealed that chlamydial infections are commonly present in Polish dairy cattle across the country.

Chlamydia gallinacea is one of the new Chlamydia species, encountered predominantly in birds and occasionally in cattle, and its dissemination, pathogenicity and zoonotic potential have not yet been fully elucidated. Until now, no case of clinical infection has been described in poultry, but the number of studies is limited. This study was conducted to evaluate the course of infection and the impact on production parameters in chicken broilers inoculated with the strain 15-56/1 isolated from a Polish flock. The presence of C. gallinacea was confirmed in oropharyngeal and cloacal swabs by real-time PCR from the fifth day post inoculation (dpi). Pathogen DNA was also detected in many internal organs of inoculated chickens. All infected animals remained asymptomatic during the entire experimental period, although statistical analyses showed that broilers in the experimental group exhibited significantly lower body weight gains and feed conversion ratios than animals in the control group. These data indicate that subclinical C. gallinacea infection in broilers may lead to financial losses for poultry farmers.

Background: Coxiella burnetiiis the etiological agent of Q fever, a zoonosis affecting many animal species including sheep and goats. The aims of this study were to evaluate the shedding ofCoxiella burnetiiin small ruminant herds and to identify the pathogen’s genotypes and sequence types (STs) using multiple-locus variable number tandem repeat analysis (MLVA) and multispacer sequence typing (MST) methods.Results: Overall, 165 samples from 43 herds of goats and 9 flocks of sheep were collected including bulk tank milk (BTM), individual milk samples, vaginal swabs, tissue sections from stillborn kids, feces and placentas. These were tested by real-time PCR targeting the IS1111element.C. burnetiiinfection was confirmed in 51.16% of the herds of goats and 22.2% of the flocks of sheep. Six out of nine samples originating from goats were successfully genotyped using the MLVA method. The presence was confirmed of two widely distributed MLVA genotypes (I and J) and genotype PL1 previously reported only in cattle. Only one sequence type (ST61) was identified; however, the majority of specimens represented partial STs and some of them may belong to ST61. Other partial STs could possibly be ST74.Conclusion: This study confirmed the relatively common occurrence ofCoxiella burnetiiin small ruminant herds in Poland. Interestingly, all genotyped samples represent cattle-associated MLVA genotypes.

Introduction: Q fever (coxiellosis) is an infectious disease of animals and humans, caused by.C. burnetii and widely distributed throughout the world. It is known that people and animals acquire the disease predominantly.via inhalation of infectious aerosols. The possibility of transmission of the pathogen by the alimentary route is still a matter of debate and remains controversial. Therefore the aim of this study was to fill the gaps in knowledge of oral transmission of.C. burnetii by conducting biological tests on the guinea pig model. Material and Methods: Guinea pigs, divided into five groups comprising a negative control and four experimental groups, received specified concentrations of.C. burnetii per os. To determine the presence of specific antibodies, blood samples were tested using CFT. Also, internal organs collected during necropsy were screened by a real-time PCR targeting I.1111. Additionally, histopathological evaluation of the tissues was performed. Results: The presence of antibodies and pathogen DNA in caecum was confirmed in one guinea pig from experimental group IV..C. burnetii was also detected in testicular tissue collected from one animal of experimental group II. Conclusions: The presence of pathogen DNA in the testicular tissue indicates that infection spreads haematogenously. In the majority of experimental animals specific antibodies and genetic material of.C. burnetii were not detected. This fact suggests that development of infection depends on many factors, such as animal immune status.

Q fever is a worldwide zoonotic disease reported in humans and many animal species including cattle. The aims of this study were to evaluate the prevalence of Coxiella (C.) burnetii shedding in Polish dairy cattle herds and to identify the pathogen’s genotypes and sequence types (STs) using multiple-locus variable number tandem repeat analysis (MLVA) and multispacer sequence typing (MST) methods. The presence of C. burnetii DNA was detected using a commercial real-time PCR kit, targeting the IS1111 element. Overall, 1,439 samples from 279 herds were tested including: 897 individual milk specimens, 101 bulk tank milk samples, 409 genital tract swabs and 32 placentas. Furthermore, 30 consumer milk samples, including 10 from vending machines and 77 dairy products were also analyzed. C. burnetii shedding was confirmed in 31.54% of tested cattle herds as well as in 69.16% of consumer milk and dairy products. Among real-time PCR–positive samples, 49 specimens obtained from 49 cattle herds and 8 samples of purchased dairy products were selected for genotyping. Overall, five previously known MLVA genotypes (I, J, BG, BE, and NM) and three new ones (proposed as PL1, PL2, and PL3) were identified. Two MST sequence types were recorded: ST16 and a novel sequence (ST61). The new genotypes and sequence types need further research particularly into their pathogenicity to humans.

Wild birds are considered as a reservoir for avian chlamydiosis posing a potential infectious threat to domestic poultry and humans. Analysis of 894 cloacal or fecal swabs from free-living birds in Poland revealed an overall Chlamydiaceae prevalence of 14.8% (n = 132) with the highest prevalence noted in Anatidae (19.7%) and Corvidae (13.4%). Further testing conducted with species-specific real-time PCR showed that 65 samples (49.2%) were positive for C. psittaci whereas only one was positive for C. avium. To classify the non-identified chlamydial agents and to genotype the C. psittaci and C. avium-positive samples, specimens were subjected to ompA-PCR and sequencing (n = 83). The ompA-based NJ dendrogram revealed that only 23 out of 83 sequences were assigned to C. psittaci, in particular to four clades representing the previously described C. psittaci genotypes B, C, Mat116 and 1V. Whereas the 59 remaining sequences were assigned to two new clades named G1 and G2, each one including sequences recently obtained from chlamydiae detected in Swedish wetland birds. G1 (18 samples from Anatidae and Rallidae) grouped closely together with genotype 1V and in relative proximity to several C. abortus isolates, and G2 (41 samples from Anatidae and Corvidae) grouped closely to C. psittaci strains of the classical ABE cluster, Matt116 and M56. Finally, deep molecular analysis of four representative isolates of genotypes 1V, G1 and G2 based on 16S rRNA, IGS and partial 23S rRNA sequences as well as MLST clearly classify these isolates within the C. abortus species. Consequently, we propose an expansion of the C. abortus species to include not only the classical isolates of mammalian origin, but also avian isolates so far referred to as atypical C. psittaci or C. psittaci/C. abortus intermediates.

Seroprevalence of Coxiella burnetii among domestic ruminants and horses in Poland