The Wood mouse (Apodemus sylvaticus) is a widespread mammalian species that acts as a reservoir host for multiple infections, including zoonotic diseases. Exposure to immunotoxins, like for instance trace metals, may reduce the ability of the host to mount proper responses to pathogens, potentially increasing the transmission and prevalence of infections. Antibody-mediated responses are crucial in preventing and limiting infections, and the quantification of the primary antibody response is considered a sensitive predictor of immunosuppression. The current study aims to investigate effects of cadmium exposure on the antibody-mediated responses of wood mice inhabiting polluted and non-polluted areas in the Netherlands. Wood mice were captured alive at different locations and immunized to sheep red blood cells (SRBC) to induce a primary antibody response. SRBC-specific antibody-producing cells, or plaque forming cells (PFC), were quantified and related to kidney cadmium levels. Differential circulating main leukocyte populations were also characterised. Cadmium concentrations in mice kidneys differed between mice captured at different locations, and increased with individual body mass, likely associated with age-related time of exposure. Effect of cadmium was apparent on the percentages of B cell counts in blood. Because of potential natural immune heterogeneity between wild rodent populations, mice immune responses were analysed and compared grouped by captured locations. Capture location had significant effect on the total counts of white blood cells. Increasing cadmium exposure in wood mice captured from polluted sites was associated with a decrease of splenic PFC counts. This field research shows that wood mice antibody responses can be impaired by cadmium exposure, even at low environmental levels, by affecting B cell functioning mainly. Impaired B cell function can make exposed mice more susceptible to infections, potentially increasing the reservoir function of their populations. It also shows that immunomodulatory effects in the field should be assessed site specifically.

The combination of emerging antibiotic resistance and lack of discovery of new antibiotic classes poses a threat to future human welfare. Antibiotics are administered to livestock at a large scale and these may enter the environment by the spreading of manure on agricultural fields. They may leach to groundwater, especially in the Netherlands which has some of the most intensive livestock farming and corresponding excessive manure spreading in the world. This study investigates the presence of antibiotics in groundwater in two regions with the most intensive livestock farming in the Netherlands. If so, the hydrochemical conditions were further elaborated. Ten multi-level wells with in total 46 filters were sampled, focusing on relatively young, previously age-dated groundwater below agricultural fields. Twenty-two antibiotics were analyzed belonging to the following antibiotic groups: tetracyclines, sulfonamides, trimethoprims, β-lactams, macrolides, lincosamides, quinolones, nitrofurans and chloramphenicol. The samples were analyzed for these antibiotics by LC-MS/MS ESI-POS/NEG (MRM) preceded by solid phase extraction which resulted in importantly low detection limits. Six antibiotics were found above detection limits in 31 filters in seven wells: sulfamethazine, sulfamethoxazole, lincomycin, chloramphenicol, ciprofloxacin, and sulfadiazine. The concentrations range from 0.3 to 18 ng L−1. Sulfonamides were detected at all measured depths down to 23 meters below surface level with apparent groundwater ages up to 40 years old. No antibiotics were detected below the nitrate/iron redox cline, which suggests that the antibiotics might undergo degradation or attenuation under nitrate-reducing redox conditions. This study provides proof that antibiotics are present in groundwater below agricultural areas in the Netherlands due to the spreading of animal manure.

Previous studies investigating health conditions of individuals living near livestock farms generally assessed short time windows. We aimed to take time-specific differences into account and to compare the prevalence of various health conditions over seven consecutive years. The sample consisted of 156,690 individuals registered in 33 general practices in a (rural) area with a high livestock density and 101,015 patients from 23 practices in other (control) areas in the Netherlands. Prevalence of health conditions were assessed using 2007–2013 electronic health record (EHR) data. Two methods were employed to assess exposure: 1) Comparisons between the study and control areas in relation to health problems, 2) Use of individual estimates of livestock exposure (in the study area) based on Geographic Information System (GIS) data. A higher prevalence of chronic bronchitis/bronchiectasis, lower respiratory tract infections and vertiginous syndrome and lower prevalence of respiratory symptoms and emphysema/COPD was found in the study area compared with the control area. A shorter distance to the nearest farm was associated with a lower prevalence of upper respiratory tract infections, respiratory symptoms, asthma, COPD/emphysema, allergic rhinitis, depression, eczema, vertiginous syndrome, dizziness and gastrointestinal infections. Especially exposure to cattle was associated with less health conditions. Living within 500m of mink farms was associated with increased chronic enteritis/ulcerative colitis. Livestock-related exposures did not seem to be an environmental risk factor for the occurrence of health conditions. Nevertheless, lower respiratory tract infections, chronic bronchitis and vertiginous syndrome were more common in the area with a high livestock density. The association between exposure to minks and chronic enteritis/ulcerative colitis remains to be elucidated.

Globally, many river sediments are seriously contaminated with persistent organic pollutants (POPs) known to accumulate in aquatic food. In the Netherlands, toxicological risks of human exposure to dioxins and dioxin-like compounds led to a ban on eel fishing in the Rhine-Meuse delta. The aim of this study is to investigate differences in serum POP levels in consumers of eel from high-polluted areas and consumers of eel from low-polluted areas or aquaculture. In total 80 Dutch men were included, aged 40–70 years, with a habitual eel consumption of at least one portion (150 g) per month. Total levels of dioxins and dioxin-like compounds were measured in serum of all participants with the DR CALUX bioassay, validated with GC-MS. For a subgroup of 38 participants extensive POP measurements were performed. We revealed that consumption of eel from polluted rivers resulted in 2.5 and up to 10 times increased levels of dioxins and polychlorinated biphenyls (PCBs) respectively compared to controls. The highest PCB levels were detected for PCB 153, with a median level of 896 ng/g lipid and a maximum level of 5000 ng/g lipid in the high-exposed group. Furthermore, hydroxylated PCB metabolites (OH-PCBs: sum of 4-OH-CB107, 4-OH-CB146, 4′-OH-CB172, and 4-OH-CB187) were 8 times higher in men who consumed eel from polluted areas, and detected at levels (median 4.5 ng/g ww) reported to cause adverse health effects. Also, the majority of the perfluoroalkyl substances (PFASs) were significantly higher in consumers of eel from pullulated areas. In conclusion, this study is the first to reveal that (past) consumption of eel from polluted rivers resulted in high body burdens of dioxins, PCBs, OH-PCBs and PFASs. We confirmed the predictions made in a former risk assessment, and the high levels of dioxins and dioxin-like compounds as well as the OH-PCBs are of health concern.

Excessive nitrogen input in natural ecosystems is a major threat to biodiversity. A coastal dune area near Amsterdam in the Netherlands suffers from high atmospheric nitrogen deposition affecting sensitive habitats such as fixed coastal dunes with herbaceous vegetation (‘grey dunes’). To mitigate its effect year round grazing was applied from 2007 until 2012. In winter, when natural food supply is low, the cattle received supplementary hay that caused additional inputs of nitrogen. Estimates based on nitrogen contents of hay, as well as of manure, showed the input through winter feeding (c. 3–14 kg N ha-1.y-1) is in the same order of magnitude as both the actual deposition (c. 17 kg N ha-1.y-1) and the critical load for a number of herbaceous habitat types (10–15 kg N ha-1.y-1). Locally, the effect of winter feeding adds to the effect of nitrogen redistribution within the area caused by the cattle's terrain usage. We conclude that winter feeding may aggravate effects of atmospheric nitrogen deposition. Keywords: Vegetation management; Manure; Nitrogen; Eutrophication; Natura 2000; Grey dune