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Background In order to evaluate the risk of human exposure to tick-borne pathogens in Belgium, a study on the prevalence of several pathogens was conducted on feeding ticks removed from humans in 2017. Methods Using a citizen science approach based on an existing notification tool for tick bites, a sample of ticks was collected across the country. Collected ticks were screened by PCR for the presence of the following pathogens: Anaplasma phagocytophilum , Babesia spp., Borrelia burgdorferi ( sensu lato ), Borrelia miyamotoi , Neoehrlichia mikurensis , Rickettsia helvetica and tick-borne encephalitis virus (TBEV). Results In total, 1599 ticks were included in the sample. The great majority of ticks belonged to Ixodes ricinus (99%); other tick species were identified as Ixodes hexagonus (0.7%) and Dermacentor reticulatus (0.3%). Borrelia burgdorferi ( s.l. ) was detected in 14% of nymphs and adult ticks. Adult ticks (20%) were more likely to be infected than nymphs (12%). The most common genospecies were B. afzelii (52%) and B. garinii (21%). Except for TBEV, the other tick-borne pathogens studied were all detected in the tick sample, although at a lower prevalence: 1.5% for Babesia spp.; 1.8% for A. phagocytophilum ; 2.4% for B. miyamotoi ; 2.8% for N. mikurensis ; and 6.8% for R. helvetica . Rickettsia raoultii , the causative agent of tick-borne lymphadenopathy, was identified for the first time in Belgium, in two out of five D. reticulatus ticks. Co-infections were found in 3.9% of the examined ticks. The most common co-infection was B. burgdorferi ( s.l .) +  N. mikurensis. Conclusions Although for most of the tick-borne diseases in Belgium, other than Lyme borreliosis, no or few cases of human infection are reported, the pathogens causing these diseases were all (except for TBEV) detected in the tick study sample. Their confirmed presence can help raise awareness among citizens and health professionals in Belgium on possible diseases other than Lyme borreliosis in patients presenting fever or other non-characteristic symptoms after a tick bite.

Background Lyme borreliosis (LB) is the most common tick-borne disease in Europe and North America, yet its economic burden remains largely unknown. This study aimed to estimate the economic cost associated with the different clinical manifestations of LB in Belgium. Methods An incidence approach and societal perspective were used to estimate the total cost-of-illness for LB in Belgium. Costs were calculated for patients with erythema migrans (EM) or disseminated/late LB, including patients who developed post-treatment Lyme disease syndrome (PTLDS). Direct medical, direct non-medical (transportation & paid help) and indirect non-medical costs (productivity losses) were included in the analysis. Ambulatory cost data were collected through a prospective cohort study from June 2016 to March 2020, in which patients with LB were followed up 6 to 12 months after diagnosis. Hospitalization costs were retrieved from the Minimal Clinical Data registry, a mandatory registry for all Belgian hospitals, linked to the Minimal Financial Data registry. Costs were expressed in 2019 euros. Results The total annual cost associated with clinical manifestations of LB in Belgium was estimated at €5.59 million (95% UI 3.82–7.98). Of these, €3.44 million (95% UI 2.05–5.48) or 62% was related to disseminated/late LB diagnoses and €2.15 million (95% UI 1.30–3.26) to EM. In general, direct medical costs and productivity losses accounted for 49.8% and 46.4% of the total costs, respectively, while direct non-medical costs accounted for only 3.8%. The estimated mean costs were €193 per EM patient and €5,148 per disseminated/late LB patient. While patients with PTLDS seemed to have somewhat higher costs compared to patients without PTLDS, the number of patients was too small to have representative estimates. Conclusions We estimate the total annual direct medical costs, direct non-medical and indirect non-medical costs associated with LB to exceed €5.5 million per year, almost evenly distributed between EM (40%) and disseminated/late LB (60%). EM costs 26 times less per patient but occurs also 16 times more frequently than disseminated/late LB. The cost burden remains limited by comparison to other infectious diseases due to the relative lower incidence.

Background Self-testing has been promoted as a means of increasing COVID-19 test coverage. In Belgium, self-testing was recommended as a complement to the formal, provider-administered indications, such as out of courtesy before meeting others and when feared to be infected. More than a year after the introduction of self-testing their place in the test strategy was evaluated. Methods We assessed trends in the number of self-tests sold, the number of positive self-tests reported, the proportion sold self-tests/total tests, and the proportion of all positive tests that were confirmed self-tests. To evaluate the reason why people use self-tests, we used the results of two online surveys among members of the general population: one among 27,397 people, held in April 2021, and one among 22,354 people, held in December 2021. Results The use of self-tests became substantial from end 2021 onwards. In the period mid-November 2021 – end-of-June 2022, the average proportion of reported sold self-tests to all COVID-19 tests was 37% and 14% of all positive tests were positive self-tests. In both surveys, the main reported reasons for using a self-test were having symptoms (34% of users in April 2021 and 31% in December 2021) and after a risk contact (27% in both April and December). Moreover, the number of self-tests sold, and the number of positive self-tests reported closely followed the same trend as the provider-administered tests in symptomatic people and high risk-contacts, which reinforces the hypothesis that they were mainly used for these two indications. Conclusions From end 2021 onwards, self-testing covered a significant part of COVID-19 testing in Belgium, which increased without doubt the testing coverage. However, the available data seem to indicate that self-testing was mostly used for indications outside of official recommendations. If and how this affected the control of the epidemic remains unknown.

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

Background Patients with Lyme borreliosis (LB) may report persisting non-specific symptoms such as fatigue, widespread musculoskeletal pain or cognitive difficulties. When present for more than 6 months and causing a reduction in daily activities, this is often referred to as post-treatment Lyme disease syndrome (PTLDS). This study aimed to compare the occurrence of symptoms between LB patients and controls, to estimate the proportion of LB patients developing PTLDS and to identify risk factors. Methods A prospective cohort study was set up including three subpopulations: patients with an erythema migrans (EM) (i) or disseminated/late LB (ii) and a non-LB control group (iii). At 6- and 12-months follow-up, the occurrence of several symptoms, including six symptoms used to define PTLDS, i.e. muscle pain, joint pain, fatigue, memory problems, difficulties concentrating and problems finding words, and impact on daily activities, was compared between LB patients and controls. Finally, the proportion of LB patients developing PTLDS as defined by the Infectious Disease Society of America was estimated, including a time frame for symptoms to be present. Results Although the risk of presenting PTLDS-related symptoms was significantly higher in EM patients (n = 120) compared to controls (n = 128) at 6 months follow-up, the risk of presenting at least one of these symptoms combined with impact on daily activities was not significantly higher in EM patients, at either 6- or 12-months follow-up. A significant association was found between disseminated/late LB (n = 15) and the occurrence of any PTLDS-symptom with an impact on daily activities at both time points. The proportion of patients with PTLDS was estimated at 5.9% (95% CI 2.7–12.9) in EM patients and 20.9% (95% CI 6.8–64.4) in patients with disseminated/late LB (RR = 3.53, 95% CI 0.98–12.68, p = 0.053). No significant risk factors were identified, which may be explained by small sample sizes. Conclusions In our study, PTLDS was present in both LB cohorts, yet with a higher percentage in disseminated/late LB patients. Additional research is needed into risk factors for and causes of this syndrome. In addition, development and validation of standardized methods to assess the PTLDS case definition, easily applicable in practice, is of great importance.

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. Graphical abstract

Background In response to the COVID-19 epidemic, caused by a novel coronavirus, it was of great importance to rapidly collect as much accurate information as possible in order to characterize the public health threat and support the health authorities in its management. Hospital-based surveillance is paramount to monitor the severity of a disease in the population. Methods Two separate surveillance systems, a Surge Capacity survey and a Clinical survey, were set up to collect complementary data on COVID-19 from Belgium’s hospitals. The Surge Capacity survey collects aggregated data to monitor the hospital capacity through occupancy rates of beds and medical devices, and to follow a set of key epidemiological indicators over time. Participation is mandatory and the daily data collection includes prevalence and incidence figures on the number of COVID-19 patients in the hospital. The Clinical survey is strongly recommended by health authorities, focusses on specific patient characteristics and relies on individual patient data provided by the hospitals at admission and discharge. Conclusions This national double-level hospital surveillance was implemented very rapidly after the first COVID-19 patients were hospitalized and revealed to be crucial to monitor hospital capacity over time and to better understand the disease in terms of risk groups and outcomes. The two approaches are complementary and serve different needs.

Tick-borne encephalitis (TBE) is a viral disease endemic in Eurasia. The virus is mainly transmitted to humans via ticks and occasionally via the consumption of unpasteurized milk products. The European Centre for Disease Prevention and Control reported an increase in TBE incidence over the past years in Europe as well as the emergence of the disease in new areas. To better understand this phenomenon, we investigated the drivers of TBE emergence and increase in incidence in humans through an expert knowledge elicitation. We listed 59 possible drivers grouped in eight domains and elicited forty European experts to: (i) allocate a score per driver, (ii) weight this score within each domain, and (iii) weight the different domains and attribute an uncertainty level per domain. An overall weighted score per driver was calculated, and drivers with comparable scores were grouped into three terminal nodes using a regression tree analysis. The drivers with the highest scores were: (i) changes in human behavior/activities; (ii) changes in eating habits or consumer demand; (iii) changes in the landscape; (iv) influence of humidity on the survival and transmission of the pathogen; (v) difficulty to control reservoir(s) and/or vector(s); (vi) influence of temperature on virus survival and transmission; (vii) number of wildlife compartments/groups acting as reservoirs or amplifying hosts; (viii) increase of autochthonous wild mammals; and (ix) number of tick species vectors and their distribution. Our results support researchers in prioritizing studies targeting the most relevant drivers of emergence and increasing TBE incidence.

Background Serological surveillance, based on the measurement of the presence of specific antibodies in a given population, can be used in addition to traditional and routine disease surveillance methods. The added value of this has been largely documented for vaccine-preventable diseases, but to a lesser extent for vector-borne diseases. This study aimed to evaluate the utility of seroprevalence data as additional source of information on the epidemiology of Lyme borreliosis in Belgium. Methods In total, 3215 residual blood samples collected in 2013–2015 were analysed with Liaison® Borrelia IgG kit (DiaSorin S.p.A, Saluggia, Italy). Positive and equivocal results were further examined with immunoblotting ( recom Line Borrelia IgG kit, Mikrogen, Neuried, Germany). Crude prevalence estimates of equivocal and seropositive results were calculated and further adjusted accounting for clustered sampling and standardized for age, sex and population per province, according to the Belgian population structure in 2014. The effect of age, sex and region on seropositivity was assessed using log-binomial regression. Results The overall weighted national seroprevalence for Borrelia burgdorferi sensu lato, adjusted for clustered sampling, age, sex and province was 1.06% (95%CI 0.67–1.67). Although not statistically significant, the highest prevalences were observed in men and in those younger than 15 years or older than 59 years of age. At provincial level, the seroprevalence estimates do not follow the geographical distribution of tick bites and diagnoses of Lyme borreliosis as detected through other surveillance systems. Conclusions Although the use of residual samples for seroprevalence estimates has several advantages, it seems to be a limited tool for serological surveillance of Lyme borreliosis in Belgium, other than follow-up of trends if repeated over time. A population-based sampling strategy might provide a more representative nationwide sample, but would be very time intensive and expensive. Seroprevalence studies within risk groups or risk areas in Belgium could provide a useful alternative approach to complement routine surveillance data of Lyme borreliosis.

Serology routinely serves as a diagnostic tool to confirm Chlamydia infections in humans. Particularly in delayed settings, such as post-outbreak scenarios where the acute phase of infection has subsided, serology is invaluable. Multiple studies, nonetheless, indicate deficiencies in specificity and sensitivity of current chlamydial antibody detection assays. Incorporation of multiple antigens per target is known to improve the accuracy of chlamydial serological assays. We, therefore, used the recomLine test (Mikrogen diagnostics) on serological samples of two cohorts, as it is the only commercially available test allowing detection of antibodies against three human pathogenic Chlamydia species (C. trachomatis, C. pneumoniae and C. psittaci) using multiple antigens per target. The first cohort (n = 156; samples collected between 2008 and 2022 during a C. trachomatis screening initiative) comprised women from the Netherlands (NL) with past exposure to C. trachomatis, while the second cohort (n = 44; samples collected in 2018 in a health examination survey) consisted of Belgian citizens (BE) with occupational or recreational exposure to chickens, representing a risk population for C. psittaci. The test indicated a statistically equivalent C. pneumoniae seroprevalence in both cohorts (39.10% in NL and 34.09% in BE; p = 0.337). As expected C. trachomatis seroprevalence was significantly higher (p < 0.001) in the Dutch cohort (48.72%), as compared to the Belgian cohort (4.55%). Lastly, C. psittaci seroprevalence did not significantly differ between the two groups (2.27% in BE and 1.92% in NL; p = 0.633), even though a higher prevalence was expected for the Belgian cohort. This prompts us to question whether the Belgian cohort truly constituted a C. psittaci risk population or whether the recomLine test is susceptible to cross-reaction of species-specific antibodies, thereby increasing C. psittaci prevalence in the Dutch cohort. We advocate for the development of affordable, highly sensitive antibody detection assays that can effectively distinguish between chlamydial species, addressing the increasing demand for enhanced serological testing methodologies.