Background:Q fever is a zoonotic disease caused by Coxiella burnetii, with domestic ruminants serving as the main reservoir. Infected animals can shed large quantities of the pathogen into the environment after normal parturition or abortion, enabling airborne transmission to humans and potentially triggering outbreaks of variable scale. Effective outbreak management requires the rapid identification of all related cases, tracing of infection source(s), and implementing proportionate control measures that protect human health while considering animal health and ecosystem integrity. Objectives:This report aimed to review and compare human Q fever outbreak management protocols across several European countries, identify areas of convergence, highlight operational challenges, and outline key gaps in diagnostic tools, source investigations, and cross-sector coordination. It also seeks to propose research priorities and recommendations to support more coordinated One Health preparedness and response. Sources:The analysis was conducted within the European Union-funded Q‑Net‑Assess project, involving partners from Belgium, France, Germany, Spain, The Netherlands, and the United Kingdom. Information was drawn from national procedures, guideline documents, and operational experiences shared by researchers and experts affiliated with academic, public health, veterinary, and reference laboratories. Content:This report describes practices related to outbreak detection, notification, case investigation, identification of sources, and implementation of control measures across countries. It maps key differences in national protocols, highlighting variability in diagnostic approaches, environmental sampling, molecular typing, and cross-sector coordination. The synthesis identifies both shared methodological elements and structural limitations that affect timely and effective outbreak management. Implications:Findings underscore the need for more harmonized and integrated Q fever outbreak management across Europe. The report outlines research priorities and provides recommendations aimed at strengthening diagnostic capacity, enhancing environmental and molecular investigation tools, and improving One Health collaboration. These insights have potential relevance for other airborne zoonotic diseases.

As part of the PRISMA project, which promotes research and innovation for productive, resilient, and healthy agro-pastoral systems in West Africa, a cross-sectional survey was conducted to assess the apparent seroprevalence and investigate epidemiological factors associated with bovine brucellosis in dairy herds. The survey was carried out in Mali (Bamako, Koulikoro, Mopti, and Sikasso regions) and Niger (Tahoua, Dosso, and Tillabéry regions). A total of 1230 animals from 82 herds were tested, including 645 animals from 43 herds in Mali and 585 animals from 39 herds in Niger. At the herd level, apparent seroprevalence was significantly higher in Mali than in Niger (37.21% vs. 15.38%). Within each country, animal-level seroprevalence varied across region, with the highest levels observed in Bamako (11.11%) and Mopti (7.62%) in Mali, and Dosso (6.67%) in Niger. High within-herd prevalence (>20%) was identified in only one herd per country. The higher overall herd-level seroprevalence in Mali was primarily driven by a greater number of herds with low (≤10%) within-herd seroprevalence. Univariate risk factor analysis at the animal level identified a borderline trend toward higher odds of seropositivity in crossbred cattle than other breeds (OR = 1.95, 95% CI: 1.00-3.81, p = 0.0503). In addition, a classification tree analysis highlighted seven exploratory signals of the positive herd status. These included four animal/management-level factors-herd region, environmental disposal of aborted placentas, the number of lactating cows (as a proxy for animal density), and reported herd abortions-as well as three human-level signals within the household: a history of miscarriage, experiencing heavy night sweats, and suffering from undulant fever. In conclusion, this survey provides new insights into bovine brucellosis and its epidemiology in dairy herds in Mali and Niger, confirming established high-prevalence areas such as Bamako and identifying new zones of concern, such as Mopti and Dosso. Our findings underscore that bovine brucellosis remains an important public health and economic threat in both countries. Given that the identified positive regions are hubs for milk production and consumption, these data may support health authorities in implementing targeted surveillance, control, and prevention strategies under a One Health approach.

Background: Bovine tuberculosis (bTB) caused by Mycobacterium bovis is a major zoonotic disease in West Africa. In Africa, bTB is endemic in cattle with a prevalence ranging from 2% up to 18%. The disease causes significant public health risks due to unpasteurized milk and milk product consumption. In the context of the EU-PRISMA project, which promotes research and innovation for productive, resilient, and healthy agropastoral systems in West Africa, a cross-sectional survey was conducted in dairy herds from Mali and Niger to assess animal, herd, and within-herd bTB prevalence, as well as to identify animal risk factors and predictors of bTB herd status. Method and principal findings: A random cross-sectional survey on dairy cattle farms using comparative intradermal tuberculin test and epidemiological inquiry was performed in four regions of Mali (Bamako, Koulikoro, Mopti, and Sikasso) and three regions of Niger (Tahoua, Dosso, and Tillabéry). Herd and animal prevalence of bTB and within-herd prevalence were significantly higher in Mali (especially in Bamako and Koulikoro) than in Niger. Several risk factors were significantly associated with animals positive to bTB, i.e., the region where animals live, the age range from 3 to 7 years old, and female animals. In addition, in regions with higher bTB prevalence, the herd with slaughtering of animals in the farm and the herd with the presence of an animal assembly area were associated with the most unfavorable status of a herd with regards to bTB. Moreover, the average and the median annual economic losses of bTB at animal level were estimated at €262 and €137 respectively, with large variability depending on the farm (between €46 and €838). Conclusion and significance: This survey provides useful data on bTB epidemiology and economical losses in Mali and Niger and urges for improvement of surveillance systems and prevention and control strategies. Cost-benefit, return of investment, or similar analyses are strongly recommended to help with decision making.

OBJECTIVES:Brucellosis in humans is mainly caused by Brucella melitensis and is associated with a risk of chronic infections and relapses. For appropriate patient treatment decisions and outcomes, clinical breakpoints as well as standard antimicrobial susceptibility testing (AST) procedures, are needed. In this study, a Europe-wide network of Brucella reference laboratories aimed, in close collaboration with the European Committee on Antimicrobial Susceptibility Testing (EUCAST), at establishing standardized AST methods, wild-type (WT) minimum inhibitory concentrations (MICs), and zone diameter distributions, and to set epidemiological cutoff (ECOFF) values for nine therapeutically relevant antimicrobial agents. METHODS:A total of 499 B. melitensis strains were tested at six study centres by broth microdilution (BMD) and disc diffusion (DD). MICs and inhibition zone diameters were curated according to EUCAST standard operating procedure (SOP) 10.2, and the results were submitted to EUCAST for ECOFFs and clinical breakpoint determination. RESULTS:BMD and DD data distributions revealed putative wild-type distributions for the tested antimicrobial agents. MIC ECOFFs were determined for all agents based on five to six distributions, encompassing 249 to 499 observations. Six isolates showed MIC values slightly above the ECOFFs, indicating the presence of a potential resistance mechanism to rifampicin, streptomycin, and trimethoprim-sulfamethoxazole. Zone diameter ECOFFs were established for rifampicin and ceftriaxone, whereas tentative (t)ECOFFs were determined for ciprofloxacin, levofloxacin, gentamicin, and streptomycin. DISCUSSION:Standardized BMD and DD methodologies for B. melitensis were validated, and AST results were used by EUCAST to set ECOFFs. Based on these, clinical breakpoints were released in v14.0 of the EUCAST clinical breakpoints table, enabling sensitive detection of resistance mechanisms and monitoring of resistance development. Genetic changes in isolates with slightly elevated MICs remain to be investigated.

Q fever is a widespread zoonotic disease caused by the bacterium, Coxiella burnetii. In ruminants, C. burnetii can cause abortions, stillbirths, premature births, and weak offspring. As part of the EU-funded Q-Net-Assess International Coordination of Research on Infectious Animal Diseases (ICRAD)-project, aimed at generating the most comprehensive understanding of C. burnetii genetic variation to date and determining the implications of this genetic variation for zoonotic risk, pathogenicity and control of C. burnetii infection, we have reviewed the protocols reported by the different project partners and/or countries to diagnose abortion in domestic ruminants. As a result of this review, we have developed guidelines for the detection of abortifacient agents in domestic ruminants, with a special focus on C. burnetii. They include a description of the essential and complementary samples needed for a definitive diagnosis, the analytical techniques to be used, and the interpretation and validity of each type of sample and technique. The most comprehensive diagnostic approach to identify an infectious agent as the cause of abortion in ruminants would include histopathology, including immunohistochemistry (IHC), on the fetus and placental membranes, complemented by bacteriology, serology, and real-time PCR analyses of different types of samples. For the specific diagnosis of C. burnetii as the causative agent of abortion, we provide guidelines based on expert opinions for the interpretation of laboratory test results in relation to their diagnostic value.

Tick-borne encephalitis (TBE) is the most frequent tick-borne viral disease transmitted by ticks in Europe and Asia. In Belgium, autochthonous cases of TBE have been reported, but even though some tick collection was carried out in the past, no TBEV-positive ticks have been found thus far. In this study, questing ticks were collected by flagging at the precise location where a patient was reported to have been bitten by a tick before developing TBE in Belgium in 2020. In total, 350 ticks were pooled by life stage (nymphs, adult females, adult males) and collection date, lysed, and RNA extracted. Quantitative reverse transcription polymerase chain reaction (RT-qPCR) was performed to detect tick-borne encephalitis virus (TBEV) and Ixodes 18S rRNA, followed by Oxford nanopore amplicon sequencing. TBEV was detected in all three types of pools. Out of 69 nymph pools, 2 were positive, in adult female pools, 2 out of 16 were positive, and 1 of the 14 adult male pools was positive. A complete sequence was retrieved through sequencing. This sequence shares greater similarity with a strain found in Finland than the neighboring Salland strain (the Netherlands) and the Neudoerfl reference strain. These findings confirm that TBE can be acquired from tick bites within the country. It is therefore necessary to increase awareness of the disease among healthcare professionals.

IntroductionQ fever, caused by Coxiella burnetii, is a disease posing significant zoonotic risk and economic impact, particularly in small ruminants. Screening prior to flock introduction is essential for disease control and prevention, especially in breeding males, given their potential role in pathogen transmission.MethodsThis study combined a survey of small ruminant farmers with a field evaluation to explore the feasibility of pre-purchase screening and to compare the performance of four diagnostic tests: real-time qPCR, antibody ELISA, intradermal testing, and an ex-vivo interferon-gamma (IFN-γ) recall assay.ResultsThe survey (n = 114) revealed widespread gaps in health status awareness and a lack of pre-screening during animal lending or borrowing, with regional differences, although most farmers supported pre-purchase screening. The field evaluation across ten farms (five positive and five negative) showed that the IFN-γ recall assay provided diagnostic accuracy comparable to ELISA (DSe: 0.80 vs. 0.74; DSp: 0.94 vs. 0.96), with higher positivity rates among unvaccinated animals (n = 122; 42.6% vs. 26.2%). The assay also proved feasible for field use, with a 24-hour antigen stimulation protocol performing equally good as the 48-hour version (AUC: 0.99 vs. 0.98). Real-time qPCR and intradermal testing showed the least utility under field conditions.DiscussionOverall, the IFN-γ recall assay emerges as a promising diagnostic tool for detecting C. burnetii infection in small ruminants, particularly in settings where vaccination history is unknown or incomplete.

Ticks are important vectors of zoonotic pathogens, and their presence can be influenced by the composition of the tick microbiome. In turn, this microbiome is shaped by environmental and ecological factors, as demonstrated in several studies conducted under controlled conditions. However, the extent of these influences under natural ecological conditions remains underexplored. In this study, we investigated the diversity of the microbiome and the prevalence of pathogens in Ixodes ricinus nymphs across three distinct Belgian ecoregions: Sandy Loam, Condroz, and Ardennes. Using real-time quantitative PCR (qPCR) and Oxford Nanopore 16S rRNA sequencing, we assessed how geography and pathogen presence influence tick-associated microbial communities. Our results revealed significant regional differences in microbiome composition and pathogen prevalence. Borrelia burgdorferi sensu lato (s.l.) was most prevalent in the Ardennes (9% (7.4-10.9) vs 3.8% (2.8-5.2) in the Condroz and 2.1% (1.4-3.2) in Sandy Loam) while Anaplasma phagocytophilum was more common in the Sandy Loam region (21.1% (18.7-23.8) vs 4% (3-5.4) in the Condroz and 3.2% (2.2-4.4) in the Ardennes). Endosymbionts such as Midichloria mitochondrii and Spiroplasma ixodetis also exhibited distinct geographic distributions. Network analysis identified potential pathogen-microbiota interactions, with certain bacterial taxa showing positive or negative associations with specific pathogens. Moreover, microbiome composition was influenced not only by ecoregion but also by microorganisms such as Rickettsia helvetica, suggesting that its colonization may actively shape microbial community structure, potentially through competition or facilitation mechanisms. Additionally, microbiome network robustness varied across ecoregions, highlighting the role of ecological context in shaping microbial interactions within ticks. These findings underscore the complex interplay between geography, pathogen presence, and microbial diversity in ticks, highlighting the importance of integrating these interactions to inform microbiome-based strategies for vector control and disease prevention.

Q fever is a widespread zoonotic disease caused by the bacterium, Coxiella burnetii. In ruminants, C. burnetii can cause abortions, stillbirths, premature births, and weak offspring. As part of the EU-funded Q-Net-Assess International Coordination of Research on Infectious Animal Diseases (ICRAD)-project, aimed at generating the most comprehensive understanding of C. burnetii genetic variation to date and determining the implications of this genetic variation for zoonotic risk, pathogenicity and control of C. burnetii infection, we have reviewed the protocols reported by the different project partners and/or countries to diagnose abortion in domestic ruminants. As a result of this review, we have developed guidelines for the detection of abortifacient agents in domestic ruminants, with a special focus on C. burnetii. They include a description of the essential and complementary samples needed for a definitive diagnosis, the analytical techniques to be used, and the interpretation and validity of each type of sample and technique. The most comprehensive diagnostic approach to identify an infectious agent as the cause of abortion in ruminants would include histopathology, including immunohistochemistry (IHC), on the fetus and placental membranes, complemented by bacteriology, serology, and real-time PCR analyses of different types of samples. For the specific diagnosis of C. burnetii as the causative agent of abortion, we provide guidelines based on expert opinions for the interpretation of laboratory test results in relation to their diagnostic value.

AIMS:The world experienced a huge number of outbreaks of highly pathogenic avian influenza (HPAI) in birds, which could represent one of the largest registered epidemics of infectious disease in food-producing animals. Therefore, mammals, including humans, are continuously exposed to HPAI viruses leading to sporadic and sometimes unusual mammal infections. The aim of this paper is to assess the risk of crossing the avian/mammalian species barrier by the currently circulating HPAI viruses, focusing on the epidemiological situation of Belgium, a representative country for Western Europe. METHODS AND RESULTS:Information on transmission pathways and species susceptibility, based on the experimental and epidemiological data, was reviewed and weighted to assess the risk of mammal infection with HPAI A(H5N1) viruses of the circulating clade 2.3.4.4b. This risk is defined as the likelihood of mammal infection by birds crossed by the clinical consequences of this infection for this animal. From the Belgian perspective, it is concluded that this risk remains 'low' to 'moderate' for captive/domestic mammal species. However, this risk was categorised as 'high' for certain species, i.e. mammals that have the opportunity to have frequent direct or indirect close contacts with infected (dead) birds, such as wild felids, wild mustelids, foxes and wild marine carnivore mammals. For some mammal species, the uncertainty associated with the assessment remains high due to an ever-changing situation. CONCLUSIONS:The longer the virus will continue to circulate in wildlife/the environment the stronger the probability of contact between infected birds and mammals will become. This will increase the related risk of viral adaptation for efficient transmission between mammal, posing concerns for public health. Regular reassessments based on the field and experimental data are therefore necessary to implement and adapt risk-based mitigation measures. This will require continuous monitoring of avian influenza viruses in both birds and mammals as well as sharing of sequence data.

BackgroundBrucellosis is a zoonosis endemic to specific geographical regions. In first line laboratories, diagnosis is made by blood culture or Rose Bengal (RB) serology.MethodsWe compare brucellosis testing between 2012-2021 at two university hospitals in Brussels, Belgium with concomitant national confirmed cases and institutional cases.ResultsRB testing increased from 30 to 211 tests/year between 2012-2021. A total of fifty-two national brucellosis cases were notified during the study period, of which fifteen cases in Brussels. No trend was noted nationally or regionally. Epidemiological data indicated travel to endemic regions, confirmed by strain testing. Institutional cases all showed symptomatic presentations with positive travel histories.ConclusionsSerologic testing inappropriately increases yearly, while annual imported brucellosis cases remain rare, and have positive travel histories and are symptomatic. We therefore support current recommendations of limiting RB testing to symptomatic patients at risk of exposure, meaning predominantly positive recent travel history.

Abstract Background Ticks carry a variety of microorganisms, some of which are pathogenic to humans. The human risk of tick-borne diseases depends on, among others, the prevalence of pathogens in ticks biting humans. To follow-up on this prevalence over time, a Belgian study from 2017 was repeated in 2021. Methods During the tick season 2021, citizens were invited to have ticks removed from their skin, send them and fill in a short questionnaire on an existing citizen science platform for the notification of tick bites (TekenNet). Ticks were morphologically identified to species and life stage level and screened using multiplex qPCR targeting, among others, Borrelia burgdorferi (sensu lato), Anaplasma phagocytophilum, Borrelia miyamotoi, Neoehrlichia mikurensis, Babesia spp., Rickettsia helvetica and tick-borne encephalitis virus (TBEV). The same methodology as in 2017 was used. Results In 2021, the same tick species as in 2017 were identified in similar proportions; of 1094 ticks, 98.7% were Ixodes ricinus, 0.8% Ixodes hexagonus and 0.5% Dermacentor reticulatus. A total of 928 nymphs and adults could be screened for the presence of pathogens. Borrelia burgdorferi (s.l.) was detected in 9.9% (95% CI 8.2–12.0%), which is significantly lower than the prevalence of 13.9% (95% CI 12.2–15.7%) in 2017 (P = 0.004). The prevalences of A. phagocytophilum (4.7%; 95% CI 3.5–6.3%) and R. helvetica (13.3%; 95% CI 11.2–15.6%) in 2021 were significantly higher compared to 2017 (1.8%; 95% CI 1.3–2.7% and 6.8%; 95% CI 5.6–8.2% respectively) (P < 0.001 for both). For the other pathogens tested, no statistical differences compared to 2017 were found, with prevalences ranging between 1.5 and 2.9% in 2021. Rickettsia raoultii was again found in D. reticulatus ticks (n = 3/5 in 2021). Similar to 2017, no TBEV was detected in the ticks. Co-infections were found in 5.1% of ticks. When combining co-infection occurrence in 2017 and 2021, a positive correlation was observed between B. burgdorferi (s.l.) and N. mikurensis and B. burgdorferi (s.l.) and B. miyamotoi (P < 0.001 for both). Conclusions Although the 2021 prevalences fell within expectations, differences were found compared to 2017. Further research to understand the explanations behind these differences is needed. Graphical Abstract

Ixodes ricinus is a vector of several pathogens of public health interest. While forests are the primary habitat for I. ricinus, its abundance and infection prevalence are expected to vary within forest stands. This study assesses the spatio-temporal variations in tick abundance and infection prevalence with three pathogens in and around a peri-urban forest where human exposure is high. Ticks were sampled multiple times in 2016 and 2018 in multiple locations with a diversity of undergrowth, using the consecutive drags method. Three zoonotic pathogens were screened for, Borrelia burgdorferi s.l., Coxiella burnetii, and Francisella tularensis. The influence of season, type of site and micro-environmental factors on tick abundance were assessed with negative binomial generalized linear mixed-effects models. We collected 1642 nymphs and 181 adult ticks. Ticks were most abundant in the spring, in warmer temperatures, and where undergrowth was higher. Sites with vegetation unaffected by human presence had higher abundance of ticks. Forest undergrowth type and height were significant predictors of the level of tick abundance in a forest. The consecutive drags method is expected to provide more precise estimates of tick abundance, presumably through more varied contacts with foliage. Borrelia burgdorferi s.l. prevalence was estimated from pooled ticks at 5.33%, C. burnetii was detected in six pools and F. tularensis was not detected. Borrelia afzelii was the dominant B. burgdorferi genospecies. Tick abundance and B. burgdorferi s.l. infection prevalence were lower than other estimates in Belgian forests.

Objectives: Bacillus anthracis clinical breakpoints, representing a systematic approach to guide clinicians in selecting the most appropriate antimicrobial treatments, are not part of the guidance from the European Committee on Antimicrobial Susceptibility Testing (EUCAST). This is because defined distributions of MIC values and of epidemiological cut-off values (ECOFFs) have been lacking. In this study, a Europe-wide network of laboratories in collaboration with EUCAST, aimed at establishing standardized antimicrobial susceptibility testing methods, wild-type MIC distributions, and ECOFFs for ten therapeutically relevant antimicrobials. Methods: About 335 B. anthracis isolates were tested by broth microdilution and disc diffusion methodologies. MIC and inhibition zone diameters were curated according to EUCAST SOP 10.2 and the results were submitted to EUCAST for ECOFFs and clinical breakpoint determination. Results: Broth microdilution and disc diffusion data distributions revealed putative wild-type distributions for the tested agents. For each antimicrobial agent, ECOFFs were defined. Three highly resistant strains with MIC values of 32 mg/L benzylpenicillin were found. MIC values slightly above the defined ECOFFs were observed in a few isolates, indicating the presence of resistance mechanisms to doxycycline, tetracycline, and amoxicillin. Discussion: B. anthracis antimicrobial susceptibility testing results were used by EUCAST to determine ECOFFs for ten antimicrobial agents. The MIC distributions were used in the process of determining clinical breakpoints. The ECOFFs can be used for the sensitive detection of isolates with resistance mechanisms, and for monitoring resistance development. Genetic changes causing phenotypic shifts in isolates displaying slightly elevated MICs remain to be investigated. Flavia Dematheis, Clin Microbiol Infect 2024;30:1170 (c) 2024 The Author(s). Published by Elsevier Ltd on behalf of European Society of Clinical Microbiology and Infectious Diseases. This is an open access article under the CC BY-NC-ND license (http:// creativecommons.org/licenses/by-nc-nd/4.0/).

Supplementary Table 4 from A Genomic Approach Reveals a Novel Mitotic Pathway in Papillomavirus Carcinogenesis

Supplementary Table 5 from A Genomic Approach Reveals a Novel Mitotic Pathway in Papillomavirus Carcinogenesis

Coxevac® is the EMA-approved veterinary vaccine for the protection of cattle and goats against Q fever, a zoonotic bacterial disease due to Coxiella burnetii. Since Coxevac® reduces bacterial shedding and clinical symptoms but does not prevent infection, novel, ready-to-use vaccine formulations are needed to increase its immunogenicity. Here, a goat vaccination-challenge model was used to evaluate the impact of the commercially available saponin-based QuilA® adjuvant on Coxevac® immunity. Upon challenge, the QuilA®-Coxevac® group showed a stronger immune response reflected in a higher magnitude of total IgG and an increase in circulating and splenic CD8+ T-cells compared to the Coxevac® and challenged-control groups. The QuilA®-Coxevac® group was characterized by a targeted Th1-type response (IFNγ, IP10) associated with increased transcripts of CD8+ and NK cells in spleens and γδ T cells in bronchial lymph nodes. Coxevac® vaccinated animals presented an intermediate expression of Th1-related genes, while the challenged-control group showed an immune response characterized by pro-inflammatory (IL1β, TNFα, IL12), Th2 (IL4 and IL13), Th17 (IL17A) and other immunoregulatory cytokines (IL6, IL10). An intriguing role was observed for γδ T cells, which were of TBX21- and SOX4-types in the QuilA®-Coxevac® and challenged control group, respectively. Overall, the addition of QuilA® resulted in a sustained Th1-type activation associated with an increased vaccine-induced bacterial clearance of 33.3% as compared to Coxevac® only. QuilA® could be proposed as a readily-applied veterinary solution to improve Coxevac® efficacy against C. burnetii infection in field settings.

This study aimed to investigate the presence of Chlamydia spp. and Parachlamydia acanthamoebae in bovine placental tissue originating from abortion and non-abortion cases in Belgium. Placentas of 164 late term bovine abortions (last trimester of gestation) and 41 non-abortion (collected after calving) cases were analysed by PCR for Chlamydia spp., Chlamydia abortus, C. psittaci and P. acanthamoebae. Additionally, a subset of 101 (75 abortion and 26 non-abortion cases) of these placenta samples were also analysed by histopathology to detect possible Chlamydia-induced lesions. In 5.4% (11/205) of the cases, Chlamydia spp. were detected, and three of those cases were positive for C. psittaci. Parachlamydia acanthamoebae was detected in 36% (75/205) of the cases, being 44% (n = 72) in abortions and 7.3% (n = 3) in non-abortions cases (p < .001). None of the cases was positive for C. abortus. Purulent and/or necrotizing placentitis with or without vasculitis was observed in 18.8% (19/101) of the histopathologically analysed placenta samples. In 5.9% (6/101) of the cases, placentitis was observed along with vasculitis. In the abortion cases, 24% (18/75) of the samples showed purulent and/or necrotizing placentitis, while purulent and/or necrotizing placentitis was visible in 3.9% (1/26) of the non-abortion cases. Placental lesions of inflammation and/or necrosis were present in 44% (15/34) of the cases where P. acanthamoebae was detected, while inflammation and/or necrosis was present in 20.9% (14/67) of the negative cases (p < .05). The detection of Chlamydia spp. and especially P. acanthamoebae, in combination with correlated histological lesions such as purulent and/or necrotizing placentitis and/or vasculitis in placental tissue following abortion, suggests a potential role of this pathogen in cases of bovine abortion in Belgium. Further in-depth studies are necessary to unravel the role of these species as abortifacient agents in cattle and to include them in bovine abortion monitoring programmes.

Illicit PDE-5 inhibitors are frequently encountered by regulatory agencies. Self-medicating with substandard and falsified (SF) PDE-5 inhibitors could be dangerous as they are likely taken without any medical supervision and might be of poor quality which could result in adverse reactions. In order to provide an overview of the quality deficiencies present in recently seized illicit PDE-5 samples that may pose health risks, we set out to identify the products’ different chemical and/or biological risks. Our results indicate that 38% of the samples harbored a chemical risk including the significant exceedance of the maximum recommended dosage, a large heterogeneity in API content between the different tablets in the same package or blister and the presence of only 40% of the claimed dosage. Moreover, our results also demonstrate that 16 of the 32 samples were not compliant with the internationally set microbiological quality standards. Startlingly, two samples were severely contaminated with potentially pathogenic bacteria, which could result in a gastrointestinal illness upon oral intake.

Background: Anaplasmosis, borreliosis, rickettsiosis and babesiosis are tick-borne diseases of medical, veterinary and economic importance. In Belgium, little is known on the prevalence of these diseases in animals and previous screenings relate only to targeted geographic regions, clinical cases or a limited number of tested samples. We therefore performed the first nationwide seroprevalence study of Anaplasma spp., A. phagocytophilum, Borrelia spp., Rickettsia spp. and Babesia spp. in Belgian cattle. We also screened questing ticks for the aforementioned pathogens. Methods: ELISAs and IFATs were performed on a representative sample set of cattle sera stratified proportionally to the number of cattle herds per province. Questing ticks were collected in areas where the highest prevalence for the forenamed pathogens in cattle serum were observed. Ticks were analyzed by quantitative PCR for A. phagocytophilum (n = 783), B. burgdorferi sensu lato (n = 783) and Rickettsia spp. (n = 715) and by PCR for Babesia spp. (n = 358). Results: The ELISA screening for antibodies to Anaplasma spp. and Borrelia spp. in cattle sera showed an overall seroprevalence of 15.6% (53/339) and 12.9% (52/402), respectively. The IFAT screening for antibodies against A. phagocytophilum, Rickettsia spp. and Babesia spp. resulted in an overall seroprevalence of 34.2% (116/339), 31.2% (99/317) and 3.4% (14/412), respectively. At the provincial level, the provinces of Liege and Walloon Brabant harboured the highest seroprevalence of Anaplasma spp. (44.4% and 42.7% respectively) and A. phagocytophilum (55.6% and 71.4%). East Flanders and Luxembourg exhibited the highest seroprevalence of Borrelia spp. (32.4%) and Rickettsia spp. (54.8%) respectively. The province of Antwerp showed the highest seroprevalence of Babesia spp. (11%). The screening of field-collected ticks resulted in a prevalence of 13.8% for B. burgdorferi s.l., with B. afzelii and B. garinii being the most common genospecies (65.7% and 17.1%, respectively). Rickettsia spp. was detected in 7.1% of the tested ticks and the only identified species was R. helvetica. A low prevalence was found for A. phagocytophilum (0.5%) and no Babesia positive tick was detected. Conclusions: The seroprevalence data in cattle indicate hot spots for tick-borne pathogens in specific provinces and highlights the importance of veterinary surveillance in anticipating the emergence of diseases among humans. The detection of all pathogens, with the exception of Babesia spp. in questing ticks, underlines the need of raising awareness among public and professionals on other tick-borne diseases along with lyme borreliosis.