From 2016 to 2020, an investigation was carried out to identify the rate of Angiostrongylus spp. infections in European badgers in Hungary. During the study, the hearts and lungs of 50 animals were dissected in order to collect adult worms, the morphometrical characteristics of which were used for species identification. PCR amplification and an 18S rDNA-sequencing analysis were also carried out. Global and local spatial autocorrelation methods were used to detect high-rated and low-rated infected animal clusters. We conducted a binary logistic regression analysis along with hierarchical agglomerative clustering to determine the relation between selected biotic and abiotic variables, and the prevalence of an A. daskalovi infection. We found a high prevalence (72%) and moderate mean intensity (14.1) of Angiostrongylus sp. infection. Morphology and sequencing revealed that all animals were infected by A. daskalovi. The results of both spatial autocorrelations suggested that the spatial distribution of infected badgers was more spatially clustered than random. The results of an analysis of the correlation between habitat characteristics and infection showed that the infected animals could be associated with dry and open landscape habitats without extended and connected canopy. It is suggested that the territorial behaviour of badgers and the landscape-directed aggregation of potential intermediate hosts might be the drivers of an A. daskalovi infection.

Several Mycoplasma species can form biofilm, facilitating their survival in the environment, and shielding them from therapeutic agents. The aim of this study was to examine the biofilm-forming ability and its potential effects on environmental survival and antibiotic resistance in Mycoplasma anserisalpingitidis, the clinically and economically most important waterfowl Mycoplasma species. The biofilm-forming ability of 32 M. anserisalpingitidis strains was examined by crystal violet assay. Biofilms and planktonic cultures of the selected strains were exposed to a temperature of 50 °C (20 and 30 min), to desiccation at room temperature (16 and 24 h), or to various concentrations of eight different antibiotics. Crystal violet staining revealed great diversity in the biofilm-forming ability of the 32 tested M. anserisalpingitidis strains, with positive staining in more than half of them. Biofilms were found to be more resistant to heat and desiccation than planktonic cultures, while no correlation was shown between biofilm formation and antibiotic susceptibility. Our results indicate that M. anserisalpingitidis biofilms may contribute to the persistence of the organisms in the environment, which should be taken into account for proper management. Antibiotic susceptibility was not affected by biofilm formation; however, it is important to note that correlations were examined only in vitro.

IntroductionMycoplasma anserisalpingitidis is one of the most important waterfowl-pathogenic mycoplasmas. Due to inadequate antibiotic treatment, many strains with high minimal inhibitory concentration (MIC) values for multiple drugs have been isolated lately. Decreased antibiotic susceptibility in several Mycoplasma species are known to be associated with mutations in topoisomerase and ribosomal genes, but other strategies such as active efflux pump mechanisms were also described. The scope of this study was the phenotypic and genetic characterization of the active efflux mechanism in M. anserisalpingitidisMethodsWe measured the MIC values in the presence and absence of different efflux pump inhibitors (EPIs), such as carbonyl cyanide m-chlorophenylhydrazine (CCCP), orthovanadate (OV), and reserpine (RSP). Moreover, bioinformatic tools were utilized to detect putative regulatory sequences of membrane transport proteins coding genes, while comparative genome analysis was performed to reveal potential markers of antibiotic resistance.ResultsOut of the three examined EPIs, CCCP decreased the MICs at least two-fold below the original MICs (in 23 cases out of 36 strains). In the presence of OV or RSP, MIC value differences could be seen only if modified dilution series (10% decrease steps were used instead of two-fold dilutions) were applied (in 24/36 cases with OV and 9/36 with RSP). During comparative genome analysis, non-synonymous single nucleotide polymorphisms (nsSNPs) were identified in genes encoding ABC membrane transport proteins, which were displayed in higher percentages in M. anserisalpingitidis strains with increased MICs. In terms of other genes, a nsSNP was identified in DNA gyrase subunit A (gyrA) gene which can be related to decreased susceptibility to enrofloxacin. The present study is the first to highlight the importance of efflux pump mechanisms in M. anserisalpingitidis.DiscussionConsidering the observed effects of the EPI CCCP against this bacterium, it can be assumed, that the use of EPIs would increase the efficiency of targeted antibiotic therapy in the future control of this pathogen. However, further research is required to obtain a more comprehensive understanding of efflux pump mechanism in this bacterium.

African swine fever (ASF) is a fatal animal disease without zoonotic potential but greatly impacts human well‐being, especially in the most vulnerable human communities. In Europe, ASF concerns mostly the wildlife domain of health. The main vector of the disease is confirmed to be the wild boar, though long‐distance jumps of the infection are due to anthropogenic effects. This study aimed to evaluate the potential role of hunting assistant personnel (beaters and carcass handlers) in ASF spread in Hungary. Based on a personal interview survey, we attempted to identify the epidemiological risk caused by hunting activities and the hunting personnel. The interviews with 58 hunting workers confirmed that an extent backyard pig sector (13 pig farmers) and pork production system (31 pork producers) existed within the study region out of the authorities’ sight. Two pig farmers did not wear special working clothes for pig caring, seven pork producers disposed of slaughter offal in the settlements periphery, and six persons regularly contacted distant pig farms. The revealed knowledge, attitude, and practice of the questioned pig farmers suggested that this sector would be very vulnerable in an epidemic situation; moreover, backyard farms would cause a great risk for wildboar populations. Considering that the study region is the third poorest region of Hungary, these findings called attention to the high epidemiologic risk of socioeconomic inequality between different regions within the European Union.

SummaryMycoplasma hyorhinisis an emerging swine pathogen bacterium with high prevalence worldwide. The main lesions caused are arthritis and polyserositis and the clinical manifestation of the disease may result in significant economic losses due to the decreased weight gain and enhanced medical costs.Our aim was to compare two challenge routes to induceM. hyorhinisinfection using the same clinical isolate. Five-week-old, Choice hybrid pigs were inoculated on two consecutive days by intravenous route (Group IV-IV) or by intravenous and intraperitoneal route (Group IV-IP). Mock infected animals were used as control (Control Group). After challenge, the clinical signs were recorded for 28 days, after which the animals were euthanized. Gross pathological and histopathological examinations, PCR detection, isolation and genotyping of the re-isolatedMycoplasmasp. and culture of bacteria other thanMycoplasmasp. were carried out. ELISA test was used to detect anti-M. hyorhinisimmunoglobulins in the sera of all animals. Pericarditis and polyarthritis were observed in both challenge groups, however the serositis was more severe in Group IV-IV. Statistically significant differences were detected between the challenged groups and the control group regarding the average daily weight gain, pathological scores and ELISA titres. Additionally, histopathological scores in Group IV-IV differed significantly from the scores in the Control Group. All re-isolated strains were the same or a close genetic variant of the original challenge strain. Our results indicate that both challenge routes are suitable for modelling the disease. However, due to the more severe pathological lesions and the more natural-like route of infection in Group IV-IV, the two-dose intravenous challenge is recommended by the authors to induce serositis and arthritis associated withM. hyorhinisinfection.