Prevalence of the causative agents of equine piroplasmosis in the South West of The Netherlands and the identification of two autochthonous clinical Theileria equi infections

Equine piroplasmosis (EP) has not been considered indigenous in The Netherlands. However, following the detection of an apparently indigenous subclinical Babesia caballi infection in a horse on Schouwen-Duiveland (an island in the Zeeland Province), a survey was undertaken between May and September 2010 to assess the prevalence of the causative agents of EP in the South-West of The Netherlands. Blood samples from 300 randomly selected horses were tested for specific antibodies against Theileria equi and B. caballi using an indirect fluorescence antibody test (IFAT), and for parasite DNA using a specific polymerase chain reaction combined with reverse line blotting (PCR-RLB). Twelve of the horses (4%) were seropositive for EP. Of these, nine (75%) were positive (titre?1:160) for B. caballi alone and three (25%) were also positive for T. equi. PCR-RLB detected T. equi DNA in five horses (1.6%), two of which were seronegative. Four (1.3%) of the positive horses (three positive for T. equi and one for both B. caballi and T. equi) were considered truly indigenous. During the study, two indigenous ponies from a farm situated outside the sampling area were diagnosed with acute clinical piroplasmosis characterized by severe anaemia and pyrexia. Blood smears showed T. equi - like inclusions in red blood cells, and T. equi infection was confirmed in both ponies by PCR-RLB. The initial subclinical B. caballi infection, the survey results and the two acute clinical EP cases confirmed the autochthonous transmission of B. caballi and T. equi infections in The Netherlands.     

Human behaviour: the key source of uncertainty in fisheries management

There is broad consensus that the main problem facing fisheries globally is too many boats chasing too few fish. Unfortunately it is also possible to argue that there are too many proposed solutions and not enough practical answers to improving fisheries management. There is a deepening divide between those who propose alternative regulatory controls on fishers, including establishing large areas permanently closed to fishing, and those who argue for better alignment of incentives combined with broad participation of resource users in fishery management decisions (in simple terms, between top down and bottom up systems of governance). However despite the choice of policy instruments used, a consistent outcome is that resource users behave in a manner that is often unintended by the designers of the management system. Hence whilst uncertainty is broadly recognized as a pervasive feature of fisheries management, to date most of the attention has focussed on only part of that uncertainty – scientific uncertainty about the status of exploited resources. The effect of uncertainty generated on the human side of fisheries science and management has received much less attention. However, the uncertainty generated by unexpected resource user behaviour is critical as it has unplanned consequences and leads to unintended management outcomes. Using empirical evidence of unexpected resource user behaviour and reviewing current responses to unexpected management outcomes, we identify different approaches that both improve prediction of human behaviour in fisheries systems and identify management measures that are more robust to these sources of uncertainty. However, unless the micro scale drivers of human behaviour that contribute to macro scale implementation uncertainty are communicated effectively to managers and considered more regularly and in greater depth, unanticipated responses to management actions will continue to undermine management systems and threaten the sustainability of fisheries.     

BSE in the Netherlands: an update

Up to 2006, there have been 82 cases of BSE in cattle born in the Netherlands. This article reviews the current situation regarding BSE in the Netherlands and summarizes the clinical symptoms of the disease. Data from the Netherlands show that a passive surveillance system, by which farmers and veterinarians have to report suspect clinical cases, has a low sensitivity. The epidemiology of, and risk factors for, BSE are discussed. All the Dutch cases of BSE can be attributed to cross-contamination of feed with meat-and-bone meal. On the basis of information about the epidemic and the cases reported to date, it is anticipated that the number of cases of BSE will continue to decline in the Netherlands and Europe. The European Commission has presented a road map that describes how the European BSE policy can be changed in the short and long term if the current favourable trend in BSE cases continues. It is time for a new phase in the management of BSE but with continued protection of the public's health and eradication of BSE.     

Changes in selected stress and immune‐related genes in Atlantic cod,Gadus morhua,following overcrowding

Using Atlantic cod, Gadus morhua, as a model for the stress response in gadoid fish, the changes in the expression of some stress and immune genes as well as the profiles of plasma cortisol were examined. Adult fish were kept at a density of ca. 100 kg m^?3 by lowering the water level in the rearing tank for 1 h and this short‐term crowding stress was repeated thrice over a 12‐h interval period. Blood samples were collected before exposure and at 2, 24 and 72 h post crowding. Plasma cortisol level significantly increased at 2 h post crowding but returned to pre‐crowding levels 24 h after exposure. The relative expression of the stress response genes, glucose transporter‐3 and a putative heat shock protein 70 significantly increased at 2 and 24 h post crowding respectively. Significant up‐regulation of the pro‐inflammatory cytokines, interleukin (IL)‐1β and IL‐8, as well as anti‐bacterial genes, g‐type lysozyme and bactericidal permeability‐increasing protein/lipopolysaccharide‐binding protein (BPI/LBP) was also observed at 2 h and the levels were maintained until 72 h post exposure, except for BPI/LBP which had maximum up‐regulation at 24 h. The present observations have implications with respect to fish welfare and assessment of the health status of the farmed fish.     

Myoblast cell interaction with polydopamine coated liposomes

Liposomes are widely used, from biosensing to drug delivery. Their coating with polymers for stability and functionalization purposes further broadens their set of relevant properties. Poly(dopamine) (PDA), a eumelanin-like material deposited via the "self"-oxidative polymerization of dopamine at mildly basic pH, has attracted considerable interest in the past few years due to its simplicity, flexibility yet fascinating properties. Herein, we characterize the coating of different types of liposomes with PDA depending on the presence of oleoyldopamine in the lipid bilayer and the dopamine hydrochloride concentration. Further, the interaction of these coated liposomes in comparison to their uncoated counterparts with myoblast cells is assessed. Their uptake/association efficiency with these cells is determined. Further, their dose-dependent cytotoxicity with and without entrapped hydrophobic cargo (thiocoraline) is characterized. Taken together, the reported results demonstrate the potential of PDA coated liposomes as a tool in biomedical applications.     

Anthogorgienes A-O, new guaiazulene-derived terpenoids from a Chinese gorgonian Anthogorgia species, and their antifouling and antibiotic activities

Fifteen new guaiazulene-based terpenoids designated anthogorgienes A-O (1-15) were isolated from a Chinese gorgonian Anthogorgia sp., together with eight known analogues (16-23). The structural patterns were classified into monomers, dimers, and trimers, which were supposed to be generated from a precursor guaiazulene and followed by side-chain and nucleus oxidation and oxidative rearrangement. The structures of new compounds were elucidated on the basis of extensive spectroscopic (IR, MS, and 1D and 2D NMR) data analysis. A possible biogenetic relationship of the isolated compounds was postulated. Some of the compounds showed potent antifouling activities against the larval settlement of barnacle Balanus amphitrite , whereas their antibiotic activities were also evaluated.     

Adherence of Actinobacillus pleuropneumoniae to primary cultures of porcine lung epithelial cells

To study adherence of Actinobacillus pleuropneumoniae to porcine lower respiratory epithelium, a cell culture model was developed using primary cultures of porcine lung epithelial cells (LEC). Adherence assays were performed and results were compared with data obtained with swine kidney cells (SK6). A. pleuropneumoniae efficiently adhered to LEC with up to 62 bacteria per cell after 2h of incubation. Reference strain of serotype 3 (R3) adhered better to LEC than reference strains of serotypes 1 (R1), 7 (R7) and 8 (R8). Overall the adherence to LEC was more rapid and up to 30-fold more efficient than adherence to SK6 cells. In search for the mechanism involved in the adherence event, we tested the effect of LPS which has previously been demonstrated to cause adherence of the pathogen to upper respiratory epithelium. Adherence assays with LPS transposon mutants demonstrated unaltered (mutant with modification in core/lipid A moiety) or even three-fold more adherence (mutants lacking O antigen) compared to the parent micro-organisms. Purified LPS of strains R1, R3, R7 and R8 did not inhibit adherence of R8 to LEC either, suggesting that LPS and particularly the O-antigen are not essential for adherence of A. pleuropneumoniae to LEC. The efficient, LPS-independent adherence of A. pleuropneumoniae to LEC cells indicates that A. pleuropneumoniae may carry different, cell type-specific adhesins and that primary cultures of lower respiratory epithelium are valuable infection models in studying A. pleuropneumoniae pathogenesis.     

In vitro markers for virulence in Yersinia ruckeri

In this study, different traits that have been associated with bacterial virulence were studied in Yersinia ruckeri. Two isolates that had been shown to cause disease and mortality in experimentally infected rainbow trout were compared with five avirulent isolates. Both virulent isolates showed high adhesion to gill and intestinal mucus of rainbow trout, whereas the majority of non‐virulent strains demonstrated significantly lower adhesion. A decrease in adherence capability following bacterial treatment with sodium metaperiodate and proteolytic enzymes suggested the involvement of carbohydrates and proteins. All strains were able to adhere to and invade chinook salmon embryo cell line (CHSE‐214), fathead minnow epithelial cell line (FHM) and rainbow trout liver cell line (R1). One non‐virulent strain was highly adhesive and invasive in the three cell lines, whereas the virulent strains showed moderate adhesive and invasive capacity. The internalization of several isolates was inhibited by colchicine and cytochalasin‐D, suggesting that microtubules and microfilaments play a role. For all strains, intracellular survival assays showed a decrease of viable bacteria in the cells 6 h after inoculation, suggesting that Y. ruckeri is not able to multiply or survive inside cultured cells. Analysis of the susceptibility to the bactericidal effect of rainbow trout serum demonstrated that virulent Y. ruckeri strains were serum resistant, whereas non‐virulent strains were generally serum sensitive.     

Pathogenesis of infectious bronchitis virus in vaccinated chickens of two different major histocompatibility B complex genotypes

Infectious bronchitis (IB) disease progression in vaccinated chickens after challenge was evaluated in a single commercial line of layer chickens presenting two different major histocompatibility complex (MHC) B complex genotypes. MHC B genotypes were determined by DNA sequence-based typing of BF2 alleles. In total, 33 B2/B15 and 47 B2/B21 chickens were vaccinated with an Ark-type IB virus (IBV) attenuated vaccine and challenged with Ark-type IBV field isolate AL/4614/98 14 days later. Additional chickens of both genotypes served as unvaccinated/challenged and unvaccinated/nonchallenged controls. Clinical signs, histopathologic analysis, detection of IBV genomes in tears, and IBV-specific immunoglobulin A (IgA) in tears were used to evaluate disease progression and immune response. The incidence of IBV respiratory signs was significantly higher in B2/21 than in B2/B15 MHC genotype birds. However, neither the severity and duration of respiratory signs nor the severity and incidence of histologic lesions differed significantly with MHC genotype. The levels of IBV-specific IgA in tears of vaccinated and challenged chickens did not differ significantly between MHC genotypes. IBV genomes were present in the tears of vaccinated and challenged birds, and the incidence of detectable IBV genomes did not vary significantly with MHC B genotype. From an applied perspective, these results indicate that vaccinated commercial outbred chickens with these MHC genotypes are equally resistant to IBV.