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Free-living amoebae often harbor obligate intracellular bacterial symbionts. Amoebophilus (A.) asiaticus is a representative of a lineage of amoeba symbionts in the phylum Bacteroidota. Here, we analyse the transcriptome of A. asiaticus strain 5a2 at four time points during its infection cycle and replication within the Acanthamoeba host using RNA sequencing. Our results reveal a dynamic transcriptional landscape throughout different A. asiaticus life cycle stages. Many intracellular bacteria and pathogens utilize eukaryotic-like proteins (ELPs) for host cell interaction and the A. asiaticus 5a2 genome shows a particularly high abundance of ELPs. We show the expression of all genes encoding ELPs and found many ELPs to be differentially expressed. At the replicative stage of A. asiaticus , ankyrin repeat proteins and tetratricopeptide/Sel1-like repeat proteins were upregulated. At the later time points, high expression levels of a type 6 secretion system that likely prepares for a new infection cycle after lysing its host, were found. This study reveals comprehensive insights into the intracellular lifestyle of A. asiaticus and highlights candidate genes for host cell interaction. The results from this study have implications for other intracellular bacteria such as other amoeba-associated bacteria and the arthropod symbionts Cardinium forming the sister lineage of A. asiaticus. [ABSTRACT FROM AUTHOR]

An economic simulation was carried out over 183 milk-producing countries to estimate the global economic impacts of 12 dairy cattle diseases and health conditions: mastitis (subclinical and clinical), lameness, paratuberculosis (Johne's disease), displaced abomasum, dystocia, metritis, milk fever, ovarian cysts, retained placenta, and ketosis (subclinical and clinical). Estimates of disease impacts on milk yield, fertility, and culling were collected from the literature, standardized, meta-analyzed using a variety of methods ranging from simple averaging to random-effects models, and adjusted for comorbidities to prevent overestimation. These comorbidity-adjusted disease impacts were then combined with a set of country-level estimates for lactational incidence or prevalence or both, herd characteristics, and price estimates within a series of Monte Carlo simulations that estimated and valued the economic losses due to these diseases. It was estimated that total annual global losses are US$65 billion (B). Subclinical ketosis, clinical mastitis, and subclinical mastitis were the costliest diseases modeled, resulting in mean annual global losses of approximately US$18B, US$13B, and US$9B, respectively. Estimated global annual losses due to clinical ketosis, displaced abomasum, dystocia, lameness, metritis, milk fever, ovarian cysts, paratuberculosis, and retained placenta were estimated to be US$0.2B, US$0.6B, US$0.6B, US$6B, US$5B, US$0.6B, US$4B, US$4B, and US$3B, respectively. Without adjustment for comorbidities, when statistical associations between diseases were disregarded, mean aggregate global losses would have been overestimated by 45%. Although annual losses were greatest in India (US$12B), the United States (US$8B), and China (US$5B), depending on the measure of losses used (losses as a percentage of gross domestic product, losses per capita, losses as a percentage of gross milk revenue), the relative economic burden of these dairy cattle diseases across countries varied markedly., (The Authors. Published by Elsevier Inc. on behalf of the American Dairy Science Association®. This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).)

This study aimed to assess the role of tourism in the spread of human immunodeficiency virus (HIV) using Malaysian epidemiological data on HIV and acquired immunodeficiency syndrome (AIDS) incidence from 1986 to 2004. A population-level mathematical model was developed with the following compartments: the population susceptible to HIV infection, the clinically confirmed HIV-positive population, the population diagnosed with AIDS, and the tourist population. Additionally, newborns infected with HIV were considered. Sensitivity analyses and variations in fixed parameter values were used to explore the effect of changes to various parameter values on HIV incidence in the model. It was determined that variations in the rate of HIV-positive inbound tourist entries and the rate of foreign tourist exits (i.e., the duration of time tourists spent in Malaysia) significantly impacted the predicted incidence of HIV and AIDS in Malaysia. The model's fit to observed HIV and AIDS incidence was evaluated, resulting in adjusted R 2 values of 53.3% and 53.2% for HIV and AIDS, respectively. Furthermore, the reproduction number (R 0 ) was also calculated to quantify the stability of the HIV endemicity in Malaysia. The findings suggest that a steady-state level of HIV in Malaysia is achievable based on the low value of R 0  = 0.0136, and the disease-free equilibrium was stable from the negative eigenvalues obtained, which is encouraging from a public health perspective. The Partial Rank Correlation Coefficient (PRCC) values between the proportion of newborns born HIV-positive, the rate of Malaysian tourist entries returning home after contracting HIV, and the rate of foreign tourist exits have a significant impact on the R 0 . The methods provide a framework for epidemiological modelling of HIV spread through transient population groups. The model results suggest that the role of tourism should not be overlooked within the set of available measures to mitigate the spread of HIV., Competing Interests: All authors declared that no competing interests exist., (© 2024 The Authors.)

Viruses are a group of widespread organisms that are often responsible for very dangerous diseases, as most of them follow a mechanism to multiply and infect their hosts as quickly as possible. Pathogen viruses also mutate regularly, with the result that measures to prevent virus transmission and recover from the disease caused are often limited. The development of new substances is very time-consuming and highly budgeted and requires the sacrifice of many living organisms. Computational chemistry methods allow faster analysis at a much lower cost and, most importantly, reduce the number of living organisms sacrificed experimentally to a minimum. Ionic liquids (ILs) are a group of chemical compounds that could potentially find a wide range of applications due to their potential virucidal activity. In our study, we conducted a complex computational analysis to predict the antiviral activity of ionic liquids against three surrogate viruses: two nonenveloped viruses, Listeria monocytogenes phage P100 and Escherichia coli phage MS2, and one enveloped virus, Pseudomonas syringae phage Phi6. Based on experimental data of toxic activity (logEC 90 ), we assigned activity classes to 154 ILs. Prediction models were created and validated according to the Organization for Economic Co-operation and Development (OECD) recommendations using the Classification Tree method. Further, we performed an external validation of our models through virtual screening on a set of 1277 theoretically generated ionic liquids and then selected 10 active ionic liquids, which were synthesized to verify their activity against the analyzed viruses. Our study proved the effectiveness and efficiency of computational methods to predict the antiviral activity of ionic liquids. Thus, computational models are a cost-effective alternative approach compared with time-consuming experimental studies where live animals are involved.

A wide variety of control and surveillance programmes that are designed and implemented based on country-specific conditions exists for infectious cattle diseases that are not regulated. This heterogeneity renders difficult the comparison of probabilities of freedom from infection estimated from collected surveillance data. The objectives of this review were to outline the methodological and epidemiological considerations for the estimation of probabilities of freedom from infection from surveillance information and review state-of-the-art methods estimating the probabilities of freedom from infection from heterogeneous surveillance data. Substantiating freedom from infection consists in quantifying the evidence of absence from the absence of evidence. The quantification usually consists in estimating the probability of observing no positive test result, in a given sample, assuming that the infection is present at a chosen (low) prevalence, called the design prevalence. The usual surveillance outputs are the sensitivity of surveillance and the probability of freedom from infection. A variety of factors influencing the choice of a method are presented; disease prevalence context, performance of the tests used, risk factors of infection, structure of the surveillance programme and frequency of testing. The existing methods for estimating the probability of freedom from infection are scenario trees, Bayesian belief networks, simulation methods, Bayesian prevalence estimation methods and the STOC free model. Scenario trees analysis is the current reference method for proving freedom from infection and is widely used in countries that claim freedom. Bayesian belief networks and simulation methods are considered extensions of scenario trees. They can be applied to more complex surveillance schemes and represent complex infection dynamics. Bayesian prevalence estimation methods and the STOC free model allow freedom from infection estimation at the herd-level from longitudinal surveillance data, considering risk factor information and the structure of the population. Comparison of surveillance outputs from heterogeneous surveillance programmes for estimating the probability of freedom from infection is a difficult task. This paper is a 'guide towards substantiating freedom from infection' that describes both all assumptions-limitations and available methods that can be applied in different settings., Competing Interests: CFa was employed by Ausvet Europe. IS-B was employed by Royal GD. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2024 Meletis, Conrady, Hopp, Lurier, Frössling, Rosendal, Faverjon, Carmo, Hodnik, Ózsvári, Kostoulas, van Schaik, Comin, Nielen, Knific, Schulz, Šerić-Haračić, Fourichon, Santman-Berends and Madouasse.)

Translocations of Rhinocerotidae are commonly performed for conservation purposes but expose the animals to a variety of stressors (e.g. prolonged fasting, confinement, novel environment, etc.). Stress may change the composition of gut microbiota, which can impact animal health and welfare. White rhinoceroses in particular can develop anorexia, diarrhea and enterocolitis after translocation. The aim of this study was to investigate the associations of age, sex and translocation on the rhinoceros' fecal bacterial microbiota composition. fecal samples were collected from rhinoceroses at capture ( n  = 16) and after a >30-hour road transport ( n  = 7). DNA was isolated from these samples and submitted for 16S rRNA V3-V4 phylotyping. Alpha diversity indices of the rhinoceros' fecal microbiota composition of different age, sex and before and after transport were compared using non-parametric statistical tests and beta diversity indices using Permutational Multivariate Analysis Of Variance (PERMANOVA). Resulting P -values were alpha-corrected ( P adj . ). Alpha and beta diversity did not differ between rhinoceroses of different age and sex. However, there was a significant difference in beta diversity between fecal samples collected from adult animals at capture and after transport. The most abundant bacterial phyla in samples collected at capture were Firmicutes and Bacteroidetes (85.76%), represented by Lachnospiraceae , Ruminococcaceae and Prevotellaceae families. The phyla Proteobacteria ( P adj . = 0.009) and Actinobacteria ( P adj . = 0.012), amongst others, increased in relative abundance from capture to after transport encompassing potentially pathogenic bacterial families such as Enterobacteriaceae ( P adj . = 0.018) and Pseudomonadaceae ( P adj . = 0.022). Important commensals such as Spirochaetes ( P adj. = 0.009), Fibrobacteres ( P adj. = 0.018) and Lachnospiraceae ( P adj. = 0.021) decreased in relative abundance. These results indicate that the stressors associated with capture and transport cause an imbalanced fecal microbiota composition in white rhinoceroses that may lead to potentially infectious intestinal disorders. This imbalance may result from recrudescence of normally innocuous pathogens, increased shedding of pathogens or increased vulnerability to new pathogens., (© The Author(s) 2023. Published by Oxford University Press and the Society for Experimental Biology.)

Since March 2021, Germany has been providing cost-free severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) antigen tests, and many day-to-day activities following the lockdown have required negative test results. Yet it remains unclear how tests have been used and whether there are patterns connected to mitigation measures. We analyzed over 50,000 anonymized records from eight test centers in a typical medium-sized city, with one of them remaining open continuously from March until December 2021. The centers exhibit distinct patterns of visitor types, with the majority tested only once in the investigated period. Individuals who underwent repeated testing tended to favor the same location. A preference for spontaneous testing grew in proportion to the availability of spare tests. Visitors aged 18 to 30 years were distinctly overrepresented compared to the local demographic. A negative binominal model showed that implemented mitigation measures had an impact on the number of tests conducted. Cost-free testing in private facilities was implemented into the German complementary screening strategy, aiming to achieve weekly population-wide testing. This study demonstrates these facilities were rarely used for regular testing but rather for meeting requirements of certified tests. The results should aid authorities in making future decisions regarding infection control., (© 2023. The Author(s).)

Objectives: To assess prevalence and types of extended-spectrum β-lactamase (ESBL)-producing bacteria in retail seafood. Methods: A literature review was completed according to international guidelines for systematic reviews, except for being performed by a single reviewer. Kruskal-Wallis and Dunn tests were used to determine statistical differences between continents or seafood types. Results: Among 12,277 hits, 42 publications from 2011 to 2023 were deemed relevant to the review's objectives. The median prevalence of ESBL-contaminated products was 19.4%. A significantly lower prevalence was observed in Europe ( p = 0.006) and Africa ( p = 0.004) compared to Asia. Amongst the 2053 isolates analyzed in the selected studies, 44.8% were ESBL-positive. The predominant type was CTX-M (93.6%), followed by TEM (6.7%) and SHV (5.0%). Only 32.6% and 18.5% of the CTX-M-positive isolates were typed to group and gene level, respectively. While group 1 (60.2%) was prevalent over group 9 (39.8%) among Enterobacterales, the opposite trend was observed in Vibrio spp. (60.0% vs. 40.0%). Information at gene level was limited to Enterobacterales, where CTX-M-15 was the most prevalent (79.2%). Conclusions: On average, one in five seafood products sold at retail globally is contaminated with ESBL-producing Enterobacterales of clinical relevance. Our findings highlight a potential risk for consumers of raw seafood, especially in Asia.

Mosquitoes of the genus Culex are important vectors of a variety of arthropod-borne viral infections. In most of the northern parts of the USA, Cx. pipiens/restuans is the predominant representative of this genus. As vectors, they play a key role in the spreading of arboviruses and thus, knowledge of the population dynamic of mosquitoes is important to understand the disease ecology of these viruses. As poikilotherm animals, the vital rates of mosquitoes are highly dependent on ambient temperature, and also on precipitation. We present a compartmental model for the population dynamics of Cx. pipiens/restuans . The model is driven by temperature, precipitation, and daytime length (which can be calculated from the geographic latitude). For model evaluation, we used long-term mosquito capture data, which were averaged from multiple sites in Cook County, Illinois. The model fitted the observation data and was able to reproduce between-year differences in the abundance of the Cx. pipiens/restuans mosquitoes, as well as the different seasonal trends. Using this model, we evaluated the effectiveness of targeting different vital rates for mosquito control strategies. The final model is able to reproduce the weekly mean Cx. pipiens/restuans abundance for Cook County with a high accuracy, and over a long time period of 20 years.

To forge a path towards livestock disease emergency preparedness in Denmark, 15 different strategies to mitigate foot-and-mouth disease (FMD) were examined by modelling epidemics initiated in cattle, pig or small ruminant herds across various production systems located in four different Danish regions (Scenario 1), or in one specific livestock production system within each of the three species geographically distributed throughout Denmark (Scenario 2). When additional mitigation strategies were implemented on top of basic control strategies in the European foot-and-mouth disease spread model (EuFMDiS), no significant benefits were predicted in terms of the number of infected farms, the epidemic control duration, and the total economic cost. Further, the model results indicated that the choice of index herd, the resources for outbreak control, and the detection time of FMD significantly influenced the course of an epidemic. The present study results emphasise the importance of basic mitigation strategies, including an effective back-and-forward traceability system, adequate resources for outbreak response, and a high level of awareness among farmers and veterinarians concerning the detection and reporting of FMD at an early stage of an outbreak for FMD control in Denmark., Competing Interests: The authors declare no conflict of interest.

Neomycin is a first-choice antibiotic for treatment of porcine enteritis caused by enterotoxigenic Escherichia coli (ETEC), but little is known about factors influencing resistance to this drug. The aims of this study were to assess antimicrobial resistance and virulence in 325 E. coli isolates obtained in 2020 from various infections in pigs, and to identify factors associated with neomycin resistance development. Susceptibility to 16 antimicrobial agents was determined by broth microdilution, and occurrence of ETEC-associated virulence factors was screened by PCR and hemolysis on blood agar. Univariate and multivariate logistic regression analyses were performed to determine if age group, virulence factors, or antibiotic use (neomycin and other antibiotics) were associated with neomycin resistance. STa, STb, LT, F4, and F18 were detected in 14%, 37%, 26%, 21% and 23% of the isolates, respectively. Resistance was low for antimicrobials of high public health importance (1.5% for cefotaxime, 1% for colistin and no fluoroquinolone resistance) but high for drugs used for treatment of ETEC enteritis (e.g. 20% for neomycin). Isolates with the ETEC pathotype were significantly associated with the weaner age group and intestinal/fecal origin. Multivariate analysis showed that recent neomycin use and presence of F4 or F18 were significantly associated with neomycin resistance amongst isolates from weaners. These results prove an association between neomycin resistance and use at the farm level. Further research is warranted to determine why neomycin resistance was associated with F4 and F18, and whether neomycin use may co-select for virulent strains., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier B.V. All rights reserved.)

During the third wave of the COVID-19 (coronavirus disease 2019) pandemic in Germany, free SARS-CoV-2 (severe acute respiratory syndrome coronavirus 2) point-of-care (PoC) antigen tests were offered to citizens at least once a week to prevent spreading by asymptomatic infected individuals. This study investigated user groups, timing, frequency, and test center locations in a typical medium-sized European city. We analyzed 27,369 pseudonymized datasets from eight centers over 12 weeks. Those were evaluated according to age, residence, appointment, and potential repeated test occurrence. The centers were visited by different groups; some centers were preferred by a predominantly younger demographic, whereas a mobile option attracted an older age group by reaching districts with few other testing possibilities. Elderly individuals were tested more spontaneously than younger individuals, and a test center at a 'park and ride' had more spontaneous visitors from outside of the city compared to other test locations. Only a small proportion of less than 4% came for testing more than five times. To preferably address many people for voluntary antigen testing, it is crucial to offer different test opportunities accounting for individual behavioral patterns, despite this requiring more complex and costly design than conventional forms.

Three cattle welfare indicators (lameness, dirtiness, and abomasal disorders) were evaluated in 412 slaughter cattle in a cross-sectional study in Austria. The aims of this study were (1) to evaluate the prevalence of lameness, dirtiness of slaughter cattle, and abomasal disorders; and (2) to determine the association between these welfare indicators and animal-related factors (e.g., housing type, carcass weight, transportation and waiting duration of the animals). The lameness prevalence was 0.73%, the abomasal disorders prevalence was 52.43%, and 88.59% of all cattle were contaminated. The latter result indicates that the cattle were kept in a dirty environment. The occurrence of abomasal disorders was associated with cattle housing systems ( p ≤ 0.00) and slaughter weight ( p = 0.03). The odds for abomasal disorders were 28.0 times higher for cattle housed on slatted flooring compared to cattle kept in a tethered system. The chance for occurrence of abomasal disorders was 3.6 times higher for cattle with a low carcass weight compared to cattle with a high carcass weight. Furthermore, significant associations were found between dirtiness (also referred to as cleanliness or contamination) and husbandry system, sex, and breed. Cattle housed in deep litter boxes had 40.8 times higher odds of being contaminated compared to cattle in a tethered housing system. Cows (odds: 32.9) and heifers (odds: 4.4) had higher odds of being contaminated with feces compared to bulls, whereby female calves (odds: 0.09) and male calves (odds: 0.02) had significantly lower odds of being contaminated. Furthermore, the breeds Brown Swiss (odds: 0.26) and Holstein-Friesian (odds: 0.14) had a significantly lower chance of being contaminated compared to Simmental cattle. Other collected factors, such as production system, transportation duration, life days of the cattle, average daily weight gain, carcass classification, and fat coverage, showed no association with the collected welfare indicators. The study presented here indicates that welfare indicators evaluated for slaughter cattle are suitable to assess cattle welfare, and improvements in husbandry may positively impact both the abomasal physiology and cleanliness of cattle.

Background: Blackleg and gas gangrene are acute clostridial infections primarily affecting cattle. The objectives of this study were to identify (i) animal-related factors influencing the occurrence and (ii) prognostic pathological findings supporting the differentiation of fatal blackleg and gas gangrene cases in the cattle population from 1998 to 2018 in Styria, Austria., Methods: Two binomial logistic models were applied to analyse the research questions. Additionally, cross-validations were performed to estimate the accuracy of the predictive models., Results: Model results show that animal-related factors (i.e., age, geographical discovery location of dead cattle, vaccination status) significantly influence the occurrence of blackleg when compared to gas gangrene. Pathological findings are similar for both diseases., Conclusions: Model results reveal that using animal-related factors has a better accuracy to predict the fatal cases caused by both pathogens. Thus, the authors recommend not relying on pathological findings as predictive factors in the differentiation between blackleg and gas gangrene in cattle., (© 2021 The Authors. Veterinary Record published by John Wiley & Sons Ltd on behalf of British Veterinary Association.)

Contingency planning allows veterinary authorities to prepare a rapid response in the event of a disease outbreak. A recently published foot-and-mouth disease (FMD) simulation study indicated concerns whether capacity was sufficient to control a potential FMD epidemic in Austria. The objectives of the study presented here were to estimate the human resources required to implement FMD control measures and to identify areas of the operational activities that could potentially delay successful control of the disease. The stochastic spatial simulation model EuFMDiS (The European Foot-and-Mouth Disease Spread Model) was used to simulate a potential FMD outbreak and its economic impact, including different control scenarios based on variations of culling, vaccination, and pre-emptive depopulation. In this context, the utilization of human resources was assessed based on the associated EuFMDiS output regarding the performance of operational activities. The assessments show that the number of personnel needed in an outbreak with a stamping-out policy would reach the peak at the end of the second week of control with a median of 540 (257-926) individuals, out of which 31% would be veterinarians. Approximately 58% of these human resources would be attributable to surveillance, followed by staff for cleaning and disinfection activities. Our analysis demonstrates that, of the operational activities, surveillance personnel were the largest factor influencing the magnitude of the outbreak. The aim of the assessment presented here is to assist veterinary authorities in the contingency planning of required human resources to respond effectively to an outbreak of animal diseases such as FMD., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Marschik, Kopacka, Stockreiter, Schmoll, Hiesel, Höflechner-Pöltl, Käsbohrer and Conrady.)

In contrast to humans and dogs, the skin microbiota of wolves is yet to be described. Here, we investigated the skin microbiota of dogs and wolves kept in outdoor packs at the Wolf Science Center (WSC) via 16S rRNA gene amplicon sequencing. Skin swab samples were also collected from human care takers and their pet dogs. When comparing the three canine groups, representing different degrees of human contact to the care takers and each other, the pet dogs showed the highest level of diversity. Additionally, while human skin was dominated by a few abundant phylotypes, the skin microbiota of the care takers who had particularly close contact with the WSC animals was more similar to the microbiota of dogs and wolves compared to the humans who had less contact with these animals. Our results suggest that domestication may have an impact on the diversity of the skin microbiota, and that the canine skin microbiota can be shared with humans, depending on the level of interaction., (© 2021. The Author(s).)

The COST action "Standardising output-based surveillance to control non-regulated diseases of cattle in the European Union (SOUND control)," aims to harmonise the results of surveillance and control programmes (CPs) for non-EU regulated cattle diseases to facilitate safe trade and improve overall control of cattle infectious diseases. In this paper we aimed to provide an overview on the diversity of control for these diseases in Europe. A non-EU regulated cattle disease was defined as an infectious disease of cattle with no or limited control at EU level, which is not included in the European Union Animal health law Categories A or B under Commission Implementing Regulation (EU) 2020/2002. A CP was defined as surveillance and/or intervention strategies designed to lower the incidence, prevalence, mortality or prove freedom from a specific disease in a region or country. Passive surveillance, and active surveillance of breeding bulls under Council Directive 88/407/EEC were not considered as CPs. A questionnaire was designed to obtain country-specific information about CPs for each disease. Animal health experts from 33 European countries completed the questionnaire. Overall, there are 23 diseases for which a CP exists in one or more of the countries studied. The diseases for which CPs exist in the highest number of countries are enzootic bovine leukosis, bluetongue, infectious bovine rhinotracheitis, bovine viral diarrhoea and anthrax (CPs reported by between 16 and 31 countries). Every participating country has on average, 6 CPs (min-max: 1-13) in place. Most programmes are implemented at a national level (86%) and are applied to both dairy and non-dairy cattle (75%). Approximately one-third of the CPs are voluntary, and the funding structure is divided between government and private resources. Countries that have eradicated diseases like enzootic bovine leukosis, bluetongue, infectious bovine rhinotracheitis and bovine viral diarrhoea have implemented CPs for other diseases to further improve the health status of cattle in their country. The control of non-EU regulated cattle diseases is very heterogenous in Europe. Therefore, the standardising of the outputs of these programmes to enable comparison represents a challenge., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Hodnik, Acinger-Rogić, Alishani, Autio, Balseiro, Berezowski, Carmo, Chaligiannis, Conrady, Costa, Cvetkovikj, Davidov, Dispas, Djadjovski, Duarte, Faverjon, Fourichon, Frössling, Gerilovych, Gethmann, Gomes, Graham, Guelbenzu, Gunn, Henry, Hopp, Houe, Irimia, Ježek, Juste, Kalaitzakis, Kaler, Kaplan, Kostoulas, Kovalenko, Kneževič, Knific, Koleci, Madouasse, Malakauskas, Mandelik, Meletis, Mincu, Mõtus, Muñoz-Gómez, Niculae, Nikitović, Ocepek, Tangen-Opsal, Ózsvári, Papadopoulos, Papadopoulos, Pelkonen, Polak, Pozzato, Rapaliuté, Ribbens, Niza-Ribeiro, Roch, Rosenbaum Nielsen, Saez, Nielsen, van Schaik, Schwan, Sekovska, Starič, Strain, Šatran, Šerić-Haračić, Tamminen, Thulke, Toplak, Tuunainen, Verner, Vilček, Yildiz and Santman-Berends.)

Some European countries have successfully implemented country-specific control programs (CPs) for infectious cattle diseases that are not regulated or are regulated only to a limited extent at the European Union (EU) level. Examples of such diseases include bovine viral diarrhea (BVD), infectious bovine rhinotracheitis (IBR), and Johne's disease (JD). The CPs vary between countries in the design and quality of collected data as well as methods used to detect infection and estimate prevalence or probability of freedom from infection. Differences in disease status between countries and non-standardized approaches to assess freedom from infection pose a risk for countries with CPs for non-regulated diseases as infected animals may influence the progress of the disease control or eradication program. The implementation of output-based standards allows estimation and comparison of the probability of freedom for non-regulated cattle diseases in European countries. The aim of the current study was to assess the existence and quality of data that could be used for estimating freedom from infection in European countries. The online data collection tool was sent to 32 countries participating in the SOUND control COST Action and was completed by 24 countries. Data on cattle demographics and data from CPs of IBR and BVD exist in more than 50% of the response countries. However, data describing risk factors and CP of JD was reported as existing in <25% of the countries. The overall quality of data in the sections on demographics and CPs of IBR and BVD were evaluated as "good", but risk factors and JD data were mostly evaluated as "fair." Data quality was considered less good mainly due to two quality criteria: accessibility and accuracy. The results of this study show that the quantity and quality of data about cattle populations and CPs are relatively similar in many surveyed countries. The outcome of this work provides an overview of the current situation in the European countries regarding data on EU non-regulated cattle diseases and will further assist in the development and implementation of output-based standards., Competing Interests: CFa was employed by the company Ausvet Europe. The remaining authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Rapaliute, van Roon, van Schaik, Santman-Berends, Koleci, Mincu, Gethmann, Conrady, Knific, Hodnik, Berezowski, Carmo, Madouasse, Tarpai, Gerilovych, Malakauskas, Sekovska, Fourichon, Kalaitzakis, Roch, Houe, Dudek, Mõtus, Ózsvári, Costa, Guelbenzu-Gonzalo, Henry, Alishani, Pozzato, Hopp, Juste, Strain, Mandelik, Vilček, Autio, Tamminen and Faverjon.)

Biofilms are formed by microorganisms protected by a self-produced matrix, most often attached to a surface. In the food processing environments biofilms endanger the product safety by the transmission of spoilage and pathogenic bacteria. In this study, we characterised the biofilm formation of the following eleven strains isolated from biofilms in a meat-processing environment: Acinetobacter harbinensis BF1, Arthrobacter sp. BF1, Brochothrix thermosphacta BF1, Carnobacterium maltaromaticum BF1, Kocuria salsicia BF1, Lactococcus piscium BF1, Microbacterium sp. BF1, Pseudomonas fragi BF1, Psychrobacter sp. BF1, Rhodococcus erythropolis BF1, Stenotrophomonas sp. BF1. We applied whole- genome sequencing and subsequent genome analysis to elucidate genetic features associated with the biofilm lifestyle. We furthermore determined the motility and studied biofilm formation on stainless steel using a static mono-species biofilm model mimicking the meat processing environment. The biomass and the EPS components carbohydrates, proteins and extracellular DNA (eDNA) of the biofilms were investigated after seven days at 10 °C. Whole-genome analysis of the isolates revealed that all strains except the Kocuria salsicia BF1 isolate, harboured biofilm associated genes, including genes for matrix production and motility. Genes involved in cellulose metabolism (present in 82% of the eleven strains) and twitching motility (present in 45%) were most frequently found. The capacity for twitching was confirmed using plate assays for all strains except Lactococcus piscium BF1, which showed the lowest motility behaviour. Differences in biofilm forming abilities could be demonstrated. The bacterial load ranged from 5.4 log CFU/cm 2 (Psychrobacter sp. isolate) to 8.7 log CFU/cm 2 (Microbacterium sp. isolate). The amount of the matrix components varied between isolates. In the biofilm of six strains we detected all three matrix components at different levels (carbohydrates, proteins and eDNA), in two only carbohydrates and eDNA, and in three only carbohydrates. Carbohydrates were detected in biofilms of all strains ranging from 0.5 to 4.3 μg glucose equivalents/cm 2 . Overall, the Microbacterium sp. strain showed the highest biofilm forming ability with high bacterial load (8.7 log CFU/cm 2 ) and high amounts of carbohydrates (2.2 μg glucose equivalents/cm 2 ), proteins (present in all experiments) and eDNA (549 ng/cm 2 ). In contrast, Brochothrix thermosphacta was a weak biofilm former, showing low bacterial load and low levels of carbohydrates in the matrix (6.2 log CFU/cm 2 and 0.5 μg glucose equivalents/cm 2 ). This study contributes to our understanding of the biofilm forming ability of bacteria highly abundant in the meat processing environment, which is crucial to develop strategies to prevent and reduce biofilm formation in the food producing environment., (Copyright © 2021 The Author(s). Published by Elsevier B.V. All rights reserved.)

The most common worldwide diarrhoea-causing agents in neonatal calves are Cryptosporidium spp. (Crypto), bovine rotavirus (BRV), bovine coronavirus (BCoV), and enterotoxigenic Escherichia coli F5 (K99) (ETEC). Crypto is a zoonotic pathogen of diarrhoea in humans, particularly for children and immunocompromised adults. Four weighted-stratified random-effect meta-analyses including meta-regression analyses were performed to calculate the worldwide mean prevalence of Crypto and associated concurrent infections with BRV, BCoV and ETEC, as well as their potential influencing factors. The meta-analysis incorporated 28 studies (56 substudies) in 17 countries that determined the presence or absence of concurrent infections with Crypto in the global calf population. Approximately half of all considered studies presented here were conducted in Europe independently of the type of infections with Crypto. Within Europe, the highest estimated mean Crypto-BRV prevalence was identified in Ireland (16.7%), the highest estimated mean Crypto-BCoV prevalence was detected in the United Kingdom (4.3%), and the highest estimated mean Crypto-ETEC prevalence across the literature was determined in Turkey (4.7%). The chance of detecting BRV, BCoV, and ETEC in calves with diarrhoea was 0.8 (confidence interval (CI): 0.6-1.0), 0.7 (CI: 0.5-1.0) and 0.6 (CI: 0.4-0.9) lower in the presence of Crypto compared to calves without Crypto. This may indicate an inhibitory effect between BRV, BCoV, ETEC, and Crypto in calves. The variance in the published prevalence across the literature can mainly be explained by the "diagnostic" factor (R 2 min-max: 0.0-40.3%), followed by the "health status of the sampled animals" (R 2 min-max: 1.4-27.3%) and "geographical region" (R 2 min-max: 5.9-23.6%).

Background: The non-regulation of animal diseases due to missing regulation at the European Union (EU) level enables member states to implement mitigation programs based on their own country-specific conditions such as priority settings of the governments, availability of financial resources, and epidemiological situation. This can result in a heterogeneous distribution of mitigation activities and prevalence levels within and/or between countries, which can cause difficulties for intracommunity trade. This article aims to describe the past, current, and future mitigation activities and associated prevalence levels for four non-regulated animal diseases, i.e., enzootic bovine leukosis (EBL), infectious bovine rhinotracheitis/infectious pustular vulvovaginitis (IBR/IPV), bovine viral diarrhea (BVD), and bluetongue disease (BT) for Austria. Over a period of 40 years (1978-2020), regulations concerning EBL, IBR/IPV, BVD, and BT were retraced to analyze the changes of legislation, focusing on sampling, testing, and mitigation activities in Austria, and were linked to the collected diagnostic testing results. The study results clearly demonstrate the adoption of the legislation by the Austrian governments in dependency of the epidemiological situations. Furthermore, our study shows that, related to the forthcoming Animal Health Law on April 21, 2021, Austria has a good initial situation to achieve disease-free status and/or free from infection status based on the current available epidemiological situation and previously implemented mitigation activities. The study results presented here are intended to contribute to a better comparison of the eradication status across the European countries for non-EU-regulated cattle diseases by providing information about the mitigation activities and data of testing results over a period of 40 years., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest., (Copyright © 2021 Roch and Conrady.)

Meat inspection data can provide valuable information about herd health to producers, veterinarians and veterinary authorities and can be used as a feedback system for farmers to improve their herd management. The aim of this study was to analyse the influence of agricultural holdings, slaughterhouses and time periods (quarters) on the occurrence and composition of the prevalence of post-mortem findings of 4 million pigs slaughtered in Austria in 2016, by applying a permutation multivariate analysis of variance. Pneumonia (21.9%) and milk spots (19.9%) were the most frequently recorded conditions. Our analysis indicated a statistically significant influence of all three considered factors (agricultural holdings, slaughterhouses and periods) on the prevalence of post-mortem findings. The observed prevalence could not only be explained by the differences between the farms of origin and slaughterhouses but also by the variability within the slaughterhouses. Much of the explained variance of the prevalence was due to differences between producers (mean R 2 = 0.61), followed by slaughterhouses (mean R 2 = 0.19) and period (mean R 2 = 0.05). To meet the demand for a valid feedback system to farmers and attending veterinarians, a robust and ideally more detailed recording of frequent pathologies, especially those affecting the respiratory tract and the liver, should be developed.

Multiple enteropathogens such as bovine rotavirus (BRV), bovine coronavirus (BCoV), Escherichia coli K99 (ETEC) and Cryptosporidium spp. (Crypto) are the most common causes of calf diarrhoea during the first 30 days of animal age. Three weighted-stratified random-effects meta-analyses were performed to calculate the worldwide prevalence of mixed infections of the causative agents (i.e., BRV-BCoV, BRV-ETEC, BRV-Crypto) and their potential influencing factors. The meta-analysis covered 41 studies (94 sub-studies) in 21 countries that determined the presence or absence of mixed infections in global calf populations. The highest worldwide estimated pooled prevalence was identified for BRV-Crypto (6.69%), followed by BRV-BCoV (2.84%), and BRV-ETEC (1.64%). The chance of detecting BCoV in calves with diarrhoea was 1.83 higher in the presence of BRV compared to calves without BRV, whereby an inhibition effect (odds ratio: 0.77) was determined between BRV and Crypto infections. The diagnostic methods were identified as a significant influencing factor in the detection of all considered mixed infections, while the other analysed factors differed in relation to their effect on prevalence. In contrast to BRV-BCoV, the prevalence of BRV-ETEC and BRV-Crypto mixed infections followed the course of individual ETEC and Crypto prevalence related to the age class of the sampled animals.