WILDbase: towards a common database to improve wildlife disease surveillance in Europe.

BackgroundTo be better prepared for emerging wildlife-borne zoonoses, we need to strengthen wildlife disease surveillance.AimThe aim of this study was to create a topical overview of zoonotic pathogens in wildlife species to identify knowledge gaps and opportunities for improvement of wildlife disease surveillance.MethodsWe created a database, which is based on a systematic literature review in Embase focused on zoonotic pathogens in 10 common urban wildlife mammals in Europe, namely brown rats, house mice, wood mice, common voles, red squirrels, European rabbits, European hedgehogs, European moles, stone martens and red foxes. In total, we retrieved 6,305 unique articles of which 882 were included.ResultsIn total, 186 zoonotic pathogen species were described, including 90 bacteria, 42 helminths, 19 protozoa, 22 viruses and 15 fungi. Most of these pathogens were only studied in one single animal species. Even considering that some pathogens are relatively species-specific, many European countries have no (accessible) data on zoonotic pathogens in these relevant animal species. We used the Netherlands as an example to show how this database can be used by other countries to identify wildlife disease surveillance gaps on a national level. Only 4% of all potential host-pathogen combinations have been studied in the Netherlands.ConclusionsThis database comprises a comprehensive overview that can guide future research on wildlife-borne zoonotic diseases both on a European and national scale. Sharing and expanding this database provides a solid starting point for future European-wide collaborations to improve wildlife disease surveillance.

Nutritional implications of feeding free-living birds in public urban areas.

Supplementary feeding can affect populations of birds. It reduces energy spent on foraging and reduces the risk of starvation, but it also increases the risk of disease transmission and predation. Supplementary feeding may reduce species richness if some species are better able to exploit supplementary food resources than others. Feeding may also artificially inflate the carrying capacity of the ecosystem, leading to bird nuisance in the form of droppings and noise. The aim of this study was to characterise and quantify the risk factors and consequences of feeding free-living birds in public areas in the western part of the city of Amsterdam. In seven study areas, the following data were collected: bird population size and species composition, feeding events, and the type and amount of supplementary food offered. Estimations were made of the nutritional energy provided and the number of birds that could be supported by the food offered. Members of the public who fed the birds were invited to complete a questionnaire on various aspects of feeding. Results showed that supplementary feeding attracts juvenile gulls and feral pigeons, which could in the long-term affect biodiversity. Bread was the main category of supplementary food being offered (estimated to be 67% of the total amount of food). The majority of respondents fed birds so as not to waste bread and meal leftovers. In six of the seven areas studied, an overabundance of nutritional energy was calculated. We conclude that the current type and extent of supplementary feeding in the city of Amsterdam is nutritionally unbalanced and affects species diversity at a local level. The overabundance is undesirable for reasons of both animal health, because it can lead to malnutrition, and public health, because surplus food attracts rats and may also have a negative effect on water quality.

Teaching Slaughter: Mapping Changes in Emotions in Veterinary Students during Training in Humane Slaughter.

As part of their training, Dutch veterinary students learn how to carry out the humane slaughter of livestock, which many students consider emotionally challenging. The aims of this study were to plot changes in self-reported emotions in veterinary students at different time points during an educational program on humane slaughter using emotion cards and to assess the change in reported emotions after adding a video and a short period of self-reflection to the program. Emotions were mapped in five groups of students at the beginning, middle, and end of the program by asking them to select from 40 cards depicting emotions in photo and text. Then two changes were made to the course program: a video of an expert slaughterman stunning and bleeding a bovine was shown, and the students were requested to spend 2 minutes picturing themselves carrying out the same procedures. To evaluate the effect of these improvements to the course, the following five groups of students were asked in the same way to indicate their emotions at the same three time points. Adding the video and short period of self-reflection did not change the emotions reported by students. Our results indicate that instruction in humane slaughter techniques involves a significant mental challenge for students. The use of emotion cards by teachers could provide useful insights into emotional aspects of the more challenging programs for students.

Cinnamaldehyde, Carvacrol and Organic Acids Affect Gene Expression of Selected Oxidative Stress and Inflammation Markers in IPEC-J2 Cells Exposed to Salmonella typhimurium.

Essential oils and organic acids are used as feed additives to improve health status and reduce colonization with pathogens. Although bactericidal in vitro, concentrations achieved in the animal gut are probably not lethal to pathogens. The aim of this study was to investigate the effects of cinnamaldehyde, carvacrol and cinnamic, lactic and propionic acids on the ability of Salmonella typhimurium ATCC 14028 (ST) to invade intestinal epithelial cells (IPEC-J2) and on the expression levels of immune related genes in the cells. The minimum inhibitory concentration (MIC) and non-inhibitory concentration (NIC) were determined and influence on the invasion capacity of ST was investigated. The structure of fimbriae and flagella was analysed by electron microscopy, and expression levels of HSP70, IkBa, IL-8 and IL-10 in the IPEC-J2 cells were carried out by q-PCR. Cinnamaldehyde, carvacrol and cinnamic and propionic acids inhibited ST invasion but not cell viability, bacterial viability and motility or the development of flagella. Propionic acid and cinnamaldehyde in combination with cinnamic acid caused structural impairment of fimbriae. Cinnamaldehyde up-regulated expression of HSP70 irrespective of the presence of organic acids or ST; exposure to carvacrol induced HSP70 only in the presence of propionic acid and ST. © 2016 The Authors. Phytotherapy Research published by John Wiley & Sons Ltd.

Iron-induced virulence of Salmonella enterica serovar typhimurium at the intestinal epithelial interface can be suppressed by carvacrol.

Oral iron therapy can increase the abundance of bacterial pathogens, e.g., Salmonella spp., in the large intestine of African children. Carvacrol is a natural compound with antimicrobial activity against various intestinal bacterial pathogens, among which is the highly prevalent Salmonella enterica serovar Typhimurium. This study aimed to explore a presumed interaction between carvacrol and bacterial iron handling and to assess the potential of carvacrol in preventing the increase of bacterial pathogenicity during high iron availability. S. Typhimurium was cultured with increasing concentrations of iron and carvacrol to study the effects of these combined interventions on growth, adhesion to intestinal epithelial cells, and iron uptake/influx in both bacterial and epithelial cells. In addition, the ability of carvacrol to remove iron from the high-affinity ligand transferrin and an Fe-dye complex was examined. Carvacrol retarded growth of S. Typhimurium at all iron conditions. Furthermore, iron-induced epithelial adhesion was effectively reduced by carvacrol at high iron concentrations. The reduction of growth and virulence by carvacrol was not paralleled by a change in iron uptake or influx into S. Typhimurium. In contrast, bioavailability of iron for epithelial cells was moderately decreased under these conditions. Further, carvacrol was shown to lack the properties of an iron binding molecule; however, it was able to weaken iron-ligand interactions by which it may possibly interfere with bacterial virulence. In conclusion, our in vitro data suggest that carvacrol has the potential to serve as a novel dietary supplement to prevent pathogenic overgrowth and colonization in the large intestine during oral iron therapy.

Synergy between essential oil components and antibiotics: a review.

With the increase in antibiotic-resistant bacteria and the lack of new antibiotics being brought onto the market, alternative strategies need to be found to cope with infections resulting from drug-resistant bacteria. A possible solution may be to combine existing antibiotics with phytochemicals to enhance the efficacy of antibiotics. A group of phytochemicals that is said to have such effects, according to in vitro studies, is essential oils (EOs) and their components. Amongst others, EOs containing carvacrol, cinnamaldehyde, cinnamic acid, eugenol and thymol can have a synergistic effect in combination with antibiotics. Several modes of action have been put forward by which antibiotics and the essential oil components may act synergistically, such as by affecting multiple targets; by physicochemical interactions and inhibiting antibacterial-resistance mechanisms. Many reported assays show additivity or moderate synergism, indicating that EOs may offer possibilities for reducing antibiotic use.

Large-scale identification of rodenticide resistance in Rattus norvegicus and Mus musculus in the Netherlands based on Vkorc1 codon 139 mutations.

BACKGROUND: Resistance to rodenticides has been reported globally and poses a considerable problem for efficacy in pest control. The most-documented resistance to rodenticides in commensal rodents is associated with mutations in the Vkorc1 gene, in particular in codon 139. Resistance to anticoagulant rodenticides has been reported in the Netherlands since 1989. A study from 2013 showed that 25% of 169 Norway rats (Rattus norvegicus) had a mutation at codon 139 of the Vkorc1 gene. To gain insight in the current status of rodenticide resistance amongst R. norvegicus and house mice Mus musculus in the Netherlands, we tested these rodents for mutations in codon 139 of the Vkorc1 gene. In addition, we collected data from pest controllers on their use of rodenticides and experience with rodenticide resistance. RESULTS: A total of 1801 rodent samples were collected throughout the country consisting of 1404 R. norvegicus and 397 M. musculus. In total, 15% of R. norvegicus [95% confidence interval (CI): 13-17%] and 38% of M. musculus (95% CI: 33-43%) carried a genetic mutation at codon 139 of the Vkorc1 gene. CONCLUSION: This study demonstrates genetic mutations at codon 139 of the Vkorc1 gene in M. musculus in the Netherlands. Resistance to anticoagulant rodenticides is present in R. norvegicus and M. musculus in multiple regions in the Netherlands. The results of this comprehensive study provide a baseline and facilitate trend analyses of Vkorc1 codon 139 mutations and evaluation of integrated pest management (IPM) strategies as these are enrolled in the Netherlands. © 2022 The Dutch Pest and Wildlife. Pest Management Science published by John Wiley & Sons Ltd on behalf of Society of Chemical Industry.

What Do They Know? Comparing Public Knowledge and Opinions about Rodent Management to the Expectations of Pest Controllers.

Integrated pest management (IPM) involves the control of pests, such as rodents, based on preventive measures and reduced use of chemical control. In view of the number of reported rodent infestations, it appears unlikely that the public has much knowledge about rodents. The objectives of this study were (i) to assess the knowledge and opinions of the public regarding prevention and control of rodent nuisance, and (ii) to assess whether pest controllers have an accurate idea of the knowledge and opinions of the public. The sample contained a total of 314 members of the public and 86 people working in the pest control sector. Responding members of the general public were asked about their knowledge and opinions about IPM in a questionnaire, whereas people working in the pest control sector were asked if they thought the general public had this knowledge and/or opinions. The results show that members of the public have a reasonable level of knowledge regarding preventive measures against rodents, which are part of IPM. People working in the pest control sector underestimate the public's knowledge of preventive measures, such as perimeter exclusion and hygiene measures. Such underestimation may affect their communication with (potential) clients.

Airborne Rodent Allergen Levels in Dutch Households: A Pilot Study.

Little research has been conducted in Europe regarding indoor exposure to airborne rodent allergens. The aims of this study were to gain insight into the prevalence of rodent allergens in airborne dust in Dutch households, to assess whether there is a relationship between rodent sightings and detectable allergens, and to identify risk factors associated with the presence of rodent allergens. Airborne dust was collected from the living rooms of 80 households distributed around central Netherlands by passive sampling using electrostatic dust collectors (EDCs). Samples were analysed for mouse (Mus m 1) and rat (Rat n 1) allergens. Participants completed a questionnaire on household and building characteristics, household pets, cleaning habits and ventilation. Mouse allergen was more prevalent than rat allergen and mouse sightings within the past year more than doubled the odds of detectable mouse allergen. Proximity to green areas, ventilation through an open window and insulation under the living room floor were determinants for detectable mouse allergen. Conversely, proximity to surface water was protective. No significant association was found between asthma and detectable mouse allergen. The passive EDC sampling method was used successfully to detect mouse and rat allergens in homes. The presence of mouse allergen was associated with previous mouse sightings. Risk factors and protective factors associated with the presence and levels of mouse allergen were identified.

Chlamydia psittaci and C. avium in feral pigeon (Columba livia domestica) droppings in two cities in the Netherlands.

BACKGROUND: Feral pigeons (Columba livia domestica) live and breed in many city centres and contact with their droppings can be a hazard for human health if the birds carry Chlamydia psittaci. OBJECTIVE: The aim of this study was to establish whether pigeon droppings in two Dutch cities (Utrecht and Haarlem) contain C. psittaci and/or C. avium, which could be a potential hazard for transmission to humans. METHODS: In May 2017 seven feral pigeon 'hot spots' with between 5 and 40+ pigeons present were identified in two cities by visual observations over two days. During the following ten days fresh droppings were collected at these hot spots and the samples were pooled per three droppings to achieve 40-41 samples per city. Samples were analysed for Chlamydia DNA with a broad range 23S Chlamydiaceae Real-Time PCR and positive samples were tested with a specific C. psittaci and C. avium Real-Time PCR. Positive C. psittaci samples were genotyped. RESULTS: C. psittaci and C. avium were detected in both cities. For C. psittaci the prevalences in Utrecht and Haarlem were 2.4% and 7.5%, respectively; for C. avium 36.6% and 20.0%, respectively. One sample contained both species. All C. psittaci samples belonged to genotype B. CONCLUSION: C. psittaci and C. avium are present in feral pigeon droppings in Utrecht and Haarlem. Human contact with droppings from infected pigeons or inhalation of dust from dried droppings represent a potential hazard to public health.

Inhibition of Salmonella enterica serotype Enteritidis on agar and raw chicken by carvacrol vapour.

Carvacrol is known to inhibit a number of food borne pathogens. The activity of carvacrol vapour was evaluated against S. enterica serotype Enteritidis on tryptone soya agar and on pieces of raw chicken. On agar, the size of the inhibition zone increased with decreasing volume of agar, increasing vapour exposure period, increasing temperature, increasing volume of carvacrol used and with decreasing bacterial density. Inhibition was equally effective under aerobic and anaerobic conditions. On chicken pieces (10 x 10 x 5 mm, UV-sterilized and inoculated with approx. 5 x 10(3) cfu) carvacrol vapour significantly reduced viable numbers of salmonellae at 4, 20 and 37 degrees C and all viable cells were eliminated by a minimum of 3 h at 37 degrees C (p<0.05). A minimum concentration of 20% carvacrol v/v in ethanol was required to achieve a significant reduction and from 40% v/v no viable cells were recovered (p<0.05). In conclusion, carvacrol vapour is effective at inhibiting the growth of S. enterica serotype Enteritidis on agar and at inhibiting and eliminating these bacteria on the surface of raw chicken.

Low temperature and binding to food components inhibit the antibacterial activity of carvacrol against Listeria monocytogenes in steak tartare.

Carvacrol is a major component of thyme and oregano essential oils and has potential uses as a food preservative. The effect of carvacrol on the growth of Listeria monocytogenes was investigated in vitro and in steak tartare. Carvacrol had strong antilisterial activity in growth medium (MIC = 1.6 mM), but no effect was observed when carvacrol was tested in steak tartare. There were two reasons for this reduced activity: the antilisterial activity of carvacrol was strongly reduced at lower temperatures (10 versus 30 degrees C), and the presence of food components interfered with the activity of carvacrol. Both bovine serum albumin and egg yolk inhibited carvacrol activity at > 0.2% (wt/vol) in growth medium. For the first time, carvacrol was found to bind to albumin, suggesting that the reduced antilisterial activity of carvacrol in foods such as dairy products and uncooked meats is the result of fewer free unbound carvacrol molecules available to interact with bacteria.

Self-reported prevalence of pests in Dutch households and the use of the health belief model to explore householders' intentions to engage in pest control.

Pests in the home are a health risk because they can be vectors for infectious disease, contribute to allergies and cause damage to buildings. The aims of this study were to record which categories of pests were reported in homes and to use a social cognition model, the health belief model, to investigate which psychological factors influence householders' intentions to control pests. An online questionnaire was completed by 413 respondents between 11 September and 31 November 2015. A large majority of respondents reported pests in or around their home within the previous year. The prevalences were: flying insects 98%, crawling insects 85%, rodents 62%, birds 58%, and moles 20%. Regression analysis for the health belief model revealed that perceiving greater benefits and fewer barriers to pest control and expecting severe consequences of zoonotic infections predicted higher intention to control pests. Intentions towards pest control were not influenced by perceiving oneself as susceptible to catching a disease from pests or health motivation (striving towards a healthy lifestyle). Intentions to engage in pest control were lower for households reporting bird prevalence. The findings suggest that interventions aimed at improving the effectiveness of domestic pest control should focus on increasing the benefits that individuals associate with effective pest control, lowering barriers, and on underlining the severity of the diseases that pests may carry.

The role of rodents in avian influenza outbreaks in poultry farms: a review.

Wild migratory birds are associated with global avian influenza virus (AIV) spread. Although direct contact with wild birds and contaminated fomites is unlikely in modern non-free range poultry farms applying biosecurity measures, AIV outbreaks still occur. This suggests involvement of other intermediate factors for virus transmission between wild birds and poultry. This review describes current evidence of the potential role of rodents in AIV transmission from wild birds to poultry and between poultry houses. Rodents can be abundant around poultry houses, share their habitat with waterfowl and can readily enter poultry houses. Survival of AIV from waterfowl in poultry house surroundings and on the coat of rodents suggests that rodents are likely to act as mechanical vector. AIVs can replicate in rodents without adaptation, resulting in high viral titres in lungs and nasal turbinates, virus presence in nasal washes and saliva, and transmission to naïve contact animals. Therefore, active AIV shedding by infected rodents may play a role in transmission to poultry. Further field and experimental studies are needed to provide evidence for a role of rodents in AIV epidemiology. Making poultry houses rodent-proof and the immediate surroundings unattractive for rodents are recommended as preventive measures against possible AIV introduction.

Structural requirements for the antimicrobial activity of carvacrol.

Carvacrol is a component of several essential oils and has been shown to exert antimicrobial activity. The structural requirements for the activity of carvacrol were determined by comparison to structurally related (nonessential oil) compounds. Removal of the aliphatic ring substituents of carvacrol slightly decreased the antimicrobial activity. The effect of the hydroxyl group of carvacrol on activity could not be determined by simply comparing it to p-cymene, because this compound is immiscible with water; therefore, 2-amino-p-cymene, the amino analogue of carvacrol, which has a similar hydrophobicity and structural characteristics, was used. 2-Amino-p-cymene had similar membrane disruption and bacterial killing characteristics as carvacrol showing that, contrary to previous reports, the hydroxyl group of carvacrol itself is not essential for the antimicrobial activity. However, the observed 3-fold lower activity for 2-amino-p-cymene as compared to carvacrol indicates special features in the antimicrobial mode of action of carvacrol due to the hydroxyl group.

Hepatitis E Virus in Farmed Rabbits, Wild Rabbits and Petting Farm Rabbits in the Netherlands.

Rabbits have been suggested as a zoonotic source of Hepatitis E virus. Phylogenetic analysis of HEV isolates from farmed, wild and pet rabbits in the Netherlands (0, 60, and 23 % respectively) showed them to be grouped amongst published rabbit HEV sequences and distinct from most human isolates. Dutch rabbits are unlikely to be a zoonotic source.

Increase in activity of essential oil components carvacrol and thymol against Escherichia coli O157:H7 by addition of food stabilizers.

The major components of oregano and thyme essential oils that had previously been shown to inhibit Escherichia coli O157:H7 were determined by high-performance liquid chromatography with UV detection and liquid chromatographic tandem mass spectrometry. The MICs and MBCs of carvacrol, thymol, p-cymene, and gamma-terpinene against a strain of E. coli O157: H7 phage type 34 isolated from bovine feces were determined by microdilution assay. The constituents were then tested in checkerboard assays to detect possible interactions. Carvacrol and thymol displayed bacteriostatic and bactericidal properties with MICs of 1.2 mmol/liter and were additive in combination. p-Cymene and gamma-terpinene displayed no measurable antibacterial activity up to 50 mmol/liter, and neither influenced the activity of carvacrol or thymol. Growth curves in the presence of nonlethal concentrations of carvacrol with the addition of agar (0.05%, wt/vol) or carrageenan (0.125%, wt/vol) as stabilizer were produced by optical density measurement. The stabilizers agar and carrageenan both significantly improved the effectiveness of carvacrol in broth, possibly because of a delay in the separation of the hydrophobic substrate from the aqueous phase of the medium. When carvacrol was dissolved in ethanol before addition to broth, stabilizers were not needed. Carvacrol and thymol, particularly when used in combination with a stabilizer or in an ethanol solution, may be effective in reducing the number or preventing growth of E. coli O157:H7 in liquid foods.

The natural antimicrobial carvacrol inhibits quorum sensing in Chromobacterium violaceum and reduces bacterial biofilm formation at sub-lethal concentrations.

The formation of biofilm by bacteria confers resistance to biocides and presents problems in medical and veterinary clinical settings. Here we report the effect of carvacrol, one of the major antimicrobial components of oregano oil, on the formation of biofilms and its activity on existing biofilms. Assays were carried out in polystyrene microplates to observe (a) the effect of 0-0.8 mM carvacrol on the formation of biofilms by selected bacterial pathogens over 24 h and (b) the effect of 0-8 mM carvacrol on the stability of pre-formed biofilms. Carvacrol was able to inhibit the formation of biofilms of Chromobacterium violaceum ATCC 12472, Salmonella enterica subsp. Typhimurium DT104, and Staphylococcus aureus 0074, while it showed no effect on formation of Pseudomonas aeruginosa (field isolate) biofilms. This inhibitory effect of carvacrol was observed at sub-lethal concentrations (<0.5 mM) where no effect was seen on total bacterial numbers, indicating that carvacrol's bactericidal effect was not causing the observed inhibition of biofilm formation. In contrast, carvacrol had (up to 8 mM) very little or no activity against existing biofilms of the bacteria described, showing that formation of the biofilm also confers protection against this compound. Since quorum sensing is an essential part of biofilm formation, the effect of carvacrol on quorum sensing of C. violaceum was also studied. Sub-MIC concentrations of carvacrol reduced expression of cviI (a gene coding for the N-acyl-L-homoserine lactone synthase), production of violacein (pigmentation) and chitinase activity (both regulated by quorum sensing) at concentrations coinciding with carvacrol's inhibiting effect on biofilm formation. These results indicate that carvacrol's activity in inhibition of biofilm formation may be related to the disruption of quorum sensing.

The natural antimicrobial carvacrol inhibits Campylobacter jejuni motility and infection of epithelial cells.

BACKGROUND: Natural compounds with anti-microbial properties are attractive reagents to reduce the use of conventional antibiotics. Carvacrol, the main constituent of oregano oil, inhibits the growth of a variety of bacterial foodborne pathogens. As concentrations of carvacrol may vary in vivo or when used in animal feed, we here investigated the effect of subinhibitory concentrations of the compound on major virulence traits of the principal bacterial foodborne pathogen Campylobacter jejuni. METHODS/PRINCIPAL FINDINGS: Motility assays revealed that subinhibitory concentrations of carvacrol inhibited the motility of C. jejuni without affecting bacterial growth. Immunoblotting and electron microscopy showed that carvacrol-treated C. jejuni still expressed flagella. The loss of motility was not caused by reduced intracellular ATP levels. In vitro infection assays demonstrated that subinhibitory concentrations of carvacrol also abolished C. jejuni invasion of human epithelial cells. Bacterial uptake of invasive Escherichia coli was not blocked by carvacrol. Exposure of C. jejuni to carvacrol prior to infection also inhibited cellular infection, indicating that the inhibition of invasion was likely caused by an effect on the bacteria rather than inhibition of epithelial cell function. CONCLUSIONS/SIGNIFICANCE: Bacterial motility and invasion of eukaryotic cells are considered key steps in C. jejuni infection. Our results indicate that subinhibitory concentrations of carvacrol effectively block these virulence traits by interfering with flagella function without disturbing intracellular ATP levels. These results broaden the spectrum of anti-microbial activity of carvacrol and support the potential of the compound for use in novel infection prevention strategies.