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Live vaccines for coccidiosis control are infrequently used in broilers, mainly due to variability in efficacy and relatively high costs. More insight in transmission of vaccine and wild-type strains can facilitate optimization of vaccination strategies and might increase its use as an alternative for anticoccidial drugs. The aim of this study was to quantify transmission of a live Eimeria acervulina vaccine strain and to determine the degree of protection against a subsequent infection with a wild-type E. acervulina strain. An experiment was carried out with 4 groups of 22 SPF broilers. At 2 days of age, 11 birds of groups 2 to 4 were vaccinated directly by oral application of E. acervulina oocysts of the Paracox™ vaccine and 11 birds were placed in contact with these birds (contact-vaccinated). Birds in group 1 remained unvaccinated (controls) and were not exposed to vaccinated birds. At day 28 of age, 6 groups of 10 birds were formed, with 2 groups (duplo) for each treatment group, i.e. vaccinated, contact-vaccinated or unvaccinated control birds. Five birds of each group were orally inoculated with wild-type E. acervulina oocysts and five were contact-exposed. Single droppings were examined daily from days 5 to 49 of age for oocyst output and to determine the time of infection. The transmission rate of the vaccine strain was estimated to be 1.6 per day and of the wild-type strain 2.3, 8.7 and 20.8 per day for vaccinated, contact-vaccinated and unvaccinated birds, respectively. Although transmission of wild-type coccidia was not significantly reduced in vaccinated or contact-vaccinated groups, both groups were equally protected against high oocyst output after infection compared to unvaccinated groups. These results suggest that factors influencing transmission of live vaccine strains in flocks may be important targets for improvement of vaccine efficacy and warrant further research., (Copyright © 2011 Elsevier Ltd. All rights reserved.)

The principal wildlife reservoir of Mycobacterium bovis in Ireland is the European badger. Studies in the Republic of Ireland (RoI) have shown that badgers culled in association with cattle herd tuberculosis breakdowns (focal culling) have a higher prevalence of infection than the badger population at large. This observation is one rationale for the medium term national strategy of focal badger culling. A vaccination strategy for the control of bovine tuberculosis (bTB) in badgers is a preferred long-term option. The Bacillus Calmette-Guérin (BCG) vaccine has been shown to decrease disease severity in captive badgers under controlled conditions. As the vaccine has been tested in a controlled environment with precise information on infection pressure, it cannot be assumed a priori that the effects of vaccination are similar in the wild, where other environmental and/or ecological factors prevail. For this reason we have designed a vaccine field trial to assess the impact of vaccination on the incidence of TB infection in a wild badger population. The selected study area for the vaccine trial (approximately 755 square kilometers) is divided into three zones each of which has similar characteristics in terms of size, number of main badger setts, cattle herds, cattle and land classification type. Three vaccination levels (100%, 50% and 0%) will be allocated to the three zones in a way that a gradient of vaccination coverage North to South is achieved. The middle zone (zone B) will be vaccinated at a 50% coverage but zone A and C will be randomly allocated with 100% or 0% vaccination coverage. Vaccination within zone B will be done randomly at individual badger level. The objective of this paper is to describe the design of a field tuberculosis vaccination trial for badgers, the epidemiological methods that were used to design the trial and the subsequent data analysis. The analysis will enable us to quantify the magnitude of the observed vaccination effect on M. bovis transmission in badgers under field conditions and to improve our knowledge of the biological effects of vaccination on susceptibility and infectiousness., (Copyright © 2011 Elsevier B.V. All rights reserved.)

Sources for human hepatitis E virus (HEV) infections of genotype 3 are largely unknown. Pigs are potential animal reservoirs for HEV. Intervention at pig farms may be desired when pigs are confirmed as a source for human infections, requiring knowledge about transmission routes. These routes are currently understudied. The current study aims to quantify the likelihood of pig feces in causing new HEV infections in pigs due to oral ingestion. We estimated the daily infection risk for pigs by modeling the fate of HEV in the fecal-oral (F-O) pathway. Using parameter values deemed most plausible by the authors based on current knowledge the daily risk of infection was 0.85 (95% interval: 0.03-1). The associated expected number of new infections per day was ∼4 (2.5% limit 0.1, the 97% limit tending to infinity) compared to 0.7 observed in a transmission experiment with pigs, and the likelihood of feces causing the transmission approached 1. In alternative scenarios, F-O transmission of HEV was also very likely to cause new infections. By reducing the total value of all explanatory variables by 2 orders of magnitude, the expected numbers of newly infected pigs approached the observed number. The likelihood of F-O transmission decreased by decreasing parameter values, allowing for at most 94% of infections being caused by additional transmission routes. Nevertheless, in all scenarios F-O transmission was estimated to contribute to HEV transmission. Thus, despite the difficulty in infecting pigs with HEV via oral inoculation, the F-O route is likely to cause HEV transmission among pigs., (© 2010 Society for Risk Analysis.)

Outbreaks of white spot syndrome virus (WSSV) in shrimp culture and the relationship between the virus and virulence are not well understood. Here, we provide evidence showing that WSSV mixed-genotype infections correlate with lower outbreak incidence and that disease outbreaks correlate with single-genotype infections. We tested 573 shrimp samples from 81 shrimp ponds in the Mekong delta with outbreak or non-outbreak status. The variable number tandem repeat (VNTR) loci of WSSV were used as molecular markers for the characterization of single- and mixed-genotype infections. The overall prevalence of mixed-genotype WSSV infections was 25.7 %. Non-outbreak ponds had a significantly higher frequency of mixed-genotype infections than outbreak ponds for all VNTR loci, both at the individual shrimp as well as at the pond level. The genetic composition of WSSV populations appears to correlate with the health status of shrimp culture in ponds. The causal relationship between genotypic diversity and disease outbreaks can now be experimentally approached.

In 2001 the epidemics of foot-and-mouth disease virus (FMDV) in Great Britain, The Netherlands and France have shown how fast FMDV may spread between farms. The massive socio-economic impact of these epidemics and the intervention measures taken demonstrate the need for quantitative assessments of the efficacy of candidate intervention strategies. Here we use a mathematical model to describe the spatial transmission of FMDV in The Netherlands and use the Dutch 2001 outbreak data to estimate model parameters. We assess the effect of ring culling strategies using a novel and fast approach producing risk maps, and discuss its consequences for ring vaccination. These risk maps identify both the geographical areas of low risk, where a given intervention strategy is likely to achieve epidemic control within only two or three farm-to-farm infection generations, and high-risk areas, where control is likely to take (much) longer. Our results indicate that certain densely populated livestock areas in the Netherlands remain high-risk areas even for strategies that extend EU minimum measures with culling or vaccination within a ring radius of several kilometres. Depending on an economic assessment, area-wide vaccination might be judged appropriate once an FMDV outbreak would have been confirmed in or close to such a high-density area. The modeling approach developed here could be readily applied to outbreak data for other diseases and in other countries., (Copyright © 2010 Elsevier B.V. All rights reserved.)

In the past decades, mathematical models have become more and more accepted as a tool to develop surveillance programs and to evaluate the efficacy of intervention measures for the control of infectious diseases such as highly pathogenic avian influenza. Predictive models are used to simulate the effect of various control measures on the course of an epidemic; analytical models are used to analyze data from outbreaks or from experiments. A key parameter in both types of models is the reproductive ratio, which indicates whether virus can be transmitted in the population, resulting in an epidemic, or not. Parameters obtained from real data using the analytical models can subsequently be used in predictive models to evaluate control strategies or surveillance programs. Examples of the use of these models are described here.

Vaccination is an attractive tool for the prevention of outbreaks of highly pathogenic avian influenza in domestic birds. It is known, however, that under certain circumstances vaccination may fail to prevent infection, and that the detection of infection in vaccinated birds can be problematic. Here, we investigate the characteristics of three serological tests (immunofluorescent antibody test (iIFAT), neuraminidase inhibition (NI) assay, and NS1 ELISA) that are able to differentiate infected from vaccinated animals. To this end, data of H7N7 infection experiments are analyzed using Bayesian methods of inference. These Bayesian methods enable validation of the tests in the absence of a gold standard, and allow one to take into account that infected birds do not always develop antibodies after infection. The results show that the N7 iIFAT and the NI assay have sensitivities for detecting antibodies of 0.95 (95% CI: 0.89-0.98) and 0.93 (95% CI: 0.78-0.99), but substantially lower sensitivities for detecting infection: 0.64 (95% CI: 0.52-0.75) and 0.63 (95% CI: 0.49-0.75). The NS1 ELISA has a low sensitivity for both detecting antibodies 0.55 (95% CI: 0.34-0.74) and infection 0.42 (95% CI: 0.28-0.56). The estimated specificities of the N7 iIFAT and the NI assay are 0.92 (95% CI: 0.87-0.95) and 0.91 (95% CI: 0.85-0.95), and 0.82 (95% CI: 0.74-0.87) for the NS1 ELISA. Additionally, our analyses suggest a strong association between the duration of virus excretion of infected birds and the probability to develop antibodies., (Copyright (c) 2009 Elsevier Ltd. All rights reserved.)

In a recent update of the Dutch contingency plan for controlling outbreaks of classical swine fever (CSF), emergency vaccination is preferred to large-scale pre-emptive culling. This policy change raised two questions: can emergency vaccination be as effective as pre-emptive culling, and what are the implications for showing freedom of infection? Here, we integrate quantitative information available on CSF virus transmission and vaccination effects into a stochastic mathematical model that describes the transmission dynamics at the level of animals, farms and livestock areas. This multilevel approach connects individual-level interventions to large-scale effects. Using this model, we compare the performance of five different control strategies applied to hypothetical CSF epidemics in The Netherlands and, for each of these strategies, we study the properties of three different screening scenarios to show freedom of infection. We find that vaccination in a ring of 2 km radius around a detected infection source is as effective as ring culling in a 1 km radius. Feasible screening scenarios, adapted to the use of emergency vaccination, can reduce the enhanced risks of (initially) undetected farm outbreaks by targeting vaccinated farms. Altogether, our results suggest that emergency vaccination against CSF can be equally effective and safe as pre-emptive culling.

To optimize control of an avian influenza outbreak knowledge of within-flock transmission is needed. This study used field data to estimate the transmission rate parameter (beta) and the influence of risk factors on within-flock transmission of highly pathogenic avian influenza (HPAI) H7N7 virus in the 2003 epidemic in The Netherlands. The estimation is based on back-calculation of daily mortality data to fit a susceptible-infectious-dead format, and these data were analysed with a generalized linear model. This back-calculation method took into account the uncertainty of the length of the latent period, the survival of an infection by some birds and the influence of farm characteristics. After analysing the fit of the different databases created by back-calculation, it could be concluded that an absence of the latency period provided the best fit. The transmission rate parameter (beta) from these field data was estimated at 4.50 per infectious chicken per day (95% CI: 2.68-7.57), which was lower than what was reported from experimental data. In contrast to general belief, none of the studied risk factors (housing system, flock size, species, age of the birds in weeks and date of depopulation) had significant influence on the estimated beta.

Foot-and-mouth disease is an extremely infectious and devastating disease affecting all species of cloven-hoofed animals. To understand the epidemiology of the causative virus and predict viral transmission dynamics, quantified transmission parameters are essential to decision makers and modellers alike. However, such quantified parameters are scarcely available, and recently a series of animal experiments was set up to obtain such data experimentally. In this communication, however, we report on the use of data from an animal experiment conducted 10 years ago to quantify transmission of foot-and-mouth disease virus between non-vaccinated sheep and from sub-clinically infected sheep to in-contact pigs. This new analysis utilises a state-of-the-art Generalised Linear Model to estimate the transmission rate. From the obtained results it is concluded that meta-analysis of "old" experiments using newly developed techniques can provide useful data to replace, reduce and refine future foot-and-mouth disease transmission experiments, thereby minimising animal suffering for research purposes.

Domestic ducks play an important role in the epidemiology of H5N1 avian influenza. Although it is known that vaccines that have a high homology with the challenge virus are able to prevent infection in ducks, little is yet known about the ability of genetically more distant vaccines in preventing infection, disease, and transmission. Here we study the effect of a widely used H5N2 vaccine (A/Chicken/Mexico/232/94/CPA) on the transmission of H5N1 virus (A/Chicken/China/1204/04) in ducks. The quantitative analyses show that despite the low level of homology between the virus and vaccine strain transmission was significantly reduced two weeks after a single or double vaccination. Mortality and disease rates were reduced markedly already one week after a single vaccination.

This study describes the results of a transmission experiment with highly pathogenic avian influenza (HPAI) H7N7 virus in 12-week-old turkeys. Cloacal and tracheal swabs as well as serum samples were taken to monitor the infection both in inoculated and in susceptible contact turkeys, which were all either unvaccinated, vaccinated once or vaccinated twice with H7N1. Swabs were tested by real-time RT-PCR and serum samples with hemagglutination inhibition test (HI). Unvaccinated contact birds had a mean infectious period of 6.2 days, and an estimated transmission rate parameter of 1.26 per infectious bird per day. However, no virus shedding was found in inoculated vaccinated turkeys and thus we concluded that vaccination with H7N1 protected against challenge with HPAI H7N7 virus.

Locally acquired hepatitis E in humans from industrialized countries has been repeatedly suggested to originate from pigs. Pigs may serve as a reservoir of hepatitis E virus (HEV) for humans when a typical infected pig causes on average more than one newly infected pig, a property that is expressed by the basic reproduction ratio R(0). In this study, R(0) for HEV transmission among pigs was estimated from chains of one-to-one transmission experiments in two blocks of five chains each. Per chain, susceptible first-generation contact pigs were contact-exposed to intravenously inoculated pigs, subsequently susceptible second-generation contact pigs were contact-exposed to infected first-generation contact pigs, and lastly, susceptible third-generation contact pigs were contact-exposed to infected second-generation contact pigs. Thus, in the second and third link of the chain, HEV-transmission due to contact with a contact-infected pig was observed. Transmission of HEV was monitored by reverse transcriptase polymerase chain reaction (RT-PCR) on individual faecal samples taken every two/three days. For susceptible pigs, the average period between exposure to an infectious pig and HEV excretion was six days (standard deviation: 4). The length of HEV-excretion (i.e. infectious period) was estimated at 49 days (95% confidence interval (CI): 17-141) for block 1 and 13 days (95% CI: 11-17) for block 2. The R0 for contact-exposure was estimated to be 8.8 (95% CI: 4-19), showing the potential of HEV to cause epidemics in populations of pigs.

Highly pathogenic avian influenza viruses can affect many bird species, with disease symptoms ranging from severe morbidity and high mortality to mild transient illness. Much is known about infections in chickens, but for other captive birds the relations between disease symptoms, excretion patterns, and transmission, as well as the effect of vaccination on these relations are not clear. We report results from experimental transmission studies with a highly pathogenic H7N7 virus and two commonly kept bird species (ringed teals and golden pheasants). The results show that depending on the host species the virus can spread in unvaccinated birds with or without disease symptoms. Vaccination reduces disease symptoms markedly, but need not always reduce virus transmission. We discuss the implications for the control of highly pathogenic avian influenza.

We investigated the influence of the spatial pattern of farms on the geographical spread of infectious livestock diseases, such as classical swine fever, foot-and-mouth disease and avian influenza in a combined analytical-numerical approach. Our purpose of this paper is to develop a method to identify the areas in which an infection has the potential to spread in an outbreak. In our model, each infected farm can infect neighbouring farms and the probability of transmission is a function of the inter-farm distance (spatial kernel). Therefore, the density of farms in an area is a good indicator for the probability of a major outbreak. In the epidemiological nomenclature, such density corresponds to a local reproduction ratio and we studied the critical behaviour of both the local density and the local reproduction ratio. We found that a threshold can be defined above which major outbreaks can occur, and the threshold value depends on the spatial kernel. Our expression for the threshold value is derived based on scaling arguments and contains two parameters in the exponents of the equation. We estimated these parameters from numerical results for the spatial spread using one particular mathematical function for the form of the spatial kernel. Subsequently, we show that our expression for the threshold using these estimated parameters agrees very well with numerical results for a number of different other functional forms of the spatial kernel (thus suggesting that we are dealing with universal parameters). As an illustration of the practical relevance of the presented method, we calculated the threshold value for avian influenza in the Netherlands and use it to produce a risk map for this disease.

A risk map of the between-farm spread of avian influenza (AI) in The Netherlands was constructed during the Dutch AI outbreak as a tool to support decision-making for control strategies. The risk map was based on an estimated distance-dependent probability of transmission of AI from an infectious farm to a susceptible farm. The estimated probability function was used to derive a threshold value for the local density of farms. In areas with local farm density above this threshold ('high risk'), transmission could lead to major outbreaks by local spread alone. In below-threshold areas ('low risk'), local spread alone will not cause a propagating epidemic. Distance-dependent probability was estimated from the data available at the start of the 2003 AI epidemic in The Netherlands. The resulting map predicted that two areas were at high risk. This approach might be applicable for other livestock infections and to other countries.

Despite continuing research efforts, knowledge of the transmission of the highly pathogenic avian influenza (HPAI) virus still has considerable gaps, which complicates epidemic control. The goal of this research was to develop a model to back-calculate the day HPAI virus is introduced into a flock, based on within-flock mortality data. The back-calculation method was based on a stochastic SEIR (susceptible (S) - latently infected (E) - infectious (I) - removed (= dead; R)) epidemic model. The latent and infectious period were assumed to be gamma distributed. Parameter values were based on experimental H7N7 within-flock transmission data. The model was used to estimate the day of virus introduction based on a defined within-flock mortality threshold (detection rule for determining AI). Our results indicate that approximately two weeks can elapse before a noticeable increase in mortality is observed after a single introduction into a flock. For example, it takes twelve (minimum 11 - maximum 15) days before AI is detected if the detection rule is fifty dead chickens on two consecutive days in a 10 000 chicken flock (current Dutch monitoring rule for notification). The results were robust for flock size and detection rule, but sensitive to the length of the latent and infectious periods. Furthermore, assuming multiple introductions on one day will result in a shorter estimated period between infection and detection. The implications of the model outcomes for detecting and tracing outbreaks of H7N7 HPAI virus are discussed.

Objective: The assessment of right ventricular (RV) size and function is important in the management of many patients with heart disease. Although magnetic resonance imaging (MRI) is considered the gold standard for quantitation of ventricular volumes and systolic function, subjective assessment ("eyeball") by echocardiography is the modality most often used for the RV. The echocardiographic "eyeball" method of assessing RV size and systolic function was compared with quantitative MRI., Design: Patients with right-sided congenital heart disease who underwent an echocardiogram within 6 months of MRI formed the study group. Four echocardiographers blinded to the MRI results reviewed the echocardiograms to subjectively assess RV size and systolic function. The reliability of an echocardiographer in accurately identifying a severely dilated RV and moderately to severely diminished RV systolic function was measured using the Kappa coefficient. Inter-rater agreement was also assessed using Kappa., Results: The study group consisted of 22 patients aged 16.6 +/- 7.1 years, with interval between MRI and echocardiogram of 49 +/- 54 days. Using echocardiography, reliability for accurately identifying a severely dilated RV was "slight" with a prevalence-adjusted bias-adjusted Kappa (PABAK) of 0.25; and for identifying moderately to severely diminished RV systolic function was fair with a PABAK of 0.43. Inter-rater agreement analysis was poor for both with Kappas of 0.07 (P = .22) and 0.12 (P = .09), respectively., Conclusion: The usefulness of the echocardiographic "eyeball" method to estimate RV size and systolic function in patients with right heart disease has limitations when compared with MRI, specifically in regard to the variability between echocardiographers.

The sex pheromone of the pistachio twig borer, Kermania pistaciella (Lepidoptera: Oinophilidae), one of the most important insect pests of pistachio, Pistacia vera, in Turkey and Iran, was identified. In gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of pheromone gland extracts of female K. pistaciella from Turkey, (2S,12Z)-2-acetoxy-12-heptadecene was identified as the major candidate pheromone component. In field experiments in Turkey, lures containing synthetic (2S,12Z)-2-acetoxy-12-heptadecene attracted large numbers of male moths. Its attractiveness was significantly reduced by the presence of the R-enantiomer or of either enantiomer of the corresponding alcohol. (2S,12Z)-2-Acetoxy-12-heptadecene is the first pheromone component identified in the Oinophilidae and the first secondary acetate pheromone component identified in the Lepidoptera.

The herd sensitivity (HSe) and herd specificity (Hsp) of clinical diagnosis of an infection with classical swine fever (CSF) virus during veterinary inspection of breeding sows in a herd was evaluated. Data gathered from visits to herds during the CSF outbreak in 1997-1998 in The Netherlands were used for the analysis. Herds were visited one or more times by the same or by different veterinarians. On the basis of the veterinarians' reports, each visit was coded as 0 (negative clinical diagnosis) or 1 (positive clinical diagnosis). The HSe for clinical diagnosis of CSF was modelled as a function of days elapsed since introduction of the virus. The moment of introduction of the CSF virus in the CSF-positive herds was unknown, so for each herd, a probability distribution for the unknown number of days since introduction was derived from serum samples collected at depopulation. The information from the reports of the veterinarians and from the test results of the serum samples at depopulation was combined in a Bayesian analysis. Data from CSF-negative herds were analysed to estimate HSp of clinical diagnosis of CSF. The HSe of clinical diagnosis was 0.5 at 37 days after virus introduction (95% CI: 31, 45) and reached 0.9 at 47 days after virus introduction (95% CI: 41, 54). The estimated herd specificity was 0.72 (95% CI: 0.64, 0.79). Dependence of HSe and HSp on characteristics of the veterinarians and the herds also was studied. Specialisation of the veterinarian significantly, although not markedly, affected the HSe.

An epidemic of high-pathogenicity avian influenza (HPAI) A virus subtype H7N7 occurred in The Netherlands in 2003 that affected 255 flocks and led to the culling of 30 million birds. To evaluate the effectiveness of the control measures, we quantified between-flock transmission characteristics of the virus in 2 affected areas, using the reproduction ratio Rh. The control measures markedly reduced the transmission of HPAI virus: Rh before detection of the outbreak in the first infected flock was 6.5 (95% confidence interval [CI], 3.1-9.9) in one area and 3.1 in another area, and it decreased to 1.2 (95% CI, 0.6-1.9) after detection of the first outbreak in both areas. The observation that Rh remained >1 suggests that the containment of the epidemic was probably due to the reduction in the number of susceptible flocks by complete depopulation of the infected areas rather than to the reduction of the transmission by the other control measures.

Veterinary vaccines are usually tested for the absence of contaminants. However, the quality control does not always imply that vaccines are not contaminated as, for example, illustrated by the bovine herpes virus 1 (BHV1) vaccine used in The Netherlands in 1999 that contained a small amount of bovine viral diarrhoea virus (BVDV1). Thousands of cows were vaccinated with BHV1 vaccine batches, and the question arose as to whether these small amounts of BVDV1, most likely not detected with in vitro tests, could have infected cattle. More in general, the question was whether the outcome of the in vitro tests, i.e. the in vitro infectivity, was indicative for the infectivity for cattle, i.e. the in vivo infectivity. We therefore carried out in vitro experiments to determine the sensitivity of a BVDV1 isolation assay. In addition, we performed two animal experiments, in which we estimated the lowest dose needed to infect calves with BVDV1. We extrapolated the experimental in vitro and in vivo results from a tissue culture infectious dose (TCID50) to a cattle infectious dose (CID50). We observed a partial response in the calves inoculated with this dose: four out of six calves turned out to be infected. In the tissue culture test, all 20 samples tested negative. The response in vivo, however, was not significantly higher than the in vitro response, which implies that no difference in susceptibility was observed between the animal test and the tissue culture test. Based on the results in our experiments, some cattle may have been infected with BVDV1 after the application of the contaminated BHV1 vaccine during the vaccination campaign. The question remains that how many cattle received contaminated vaccine, and became infected with BVDV1.

The foot-and-mouth disease outbreak in The Netherlands in 2001 most likely started on a mixed veal-calf/dairy-goat farm. The outbreak among the 74 calves on this farm appeared to be limited to four animals, and no clinical signs of FMD were reported. Also on a second veal-calf farm minor clinical signs and limited virus transmission were observed. Since FMD is known to be a very contagious disease, and can cause severe lesions, these observations were disputed. Therefore, we carried out two experiments to determine whether the Dutch FMD virus isolate from 2001 does spread among individually housed calves with limited contacts, either indirect (experiment 1) or direct (experiment 2). In experiment 1, four pairs of calves were housed in an individual box at 1m distance from each other. In experiment 2, two groups of three calves were housed in individual boxes, directly bordering each other. We infected one animal per pair in experiment 1, and the calf in the middle in experiment 2. We recorded clinical signs, virus shedding in saliva and the development of antibodies. In addition, we determined whether the virus was transmitted from the inoculated calves to the neighbour(s). All inoculated calves showed mild signs of FMD--fever, and some vesicles on hooves and/or in the mouth--but only one calf showed signs that were visible without physical examination. All inoculated calves shed virus in the saliva and developed neutralising antibodies. None of the contact animals seroconverted, indicating that virus transmission did not occur. These experiments showed that no virus transmission among individual housed calves can occur. This finding supports the hypothesis of the route of virus introduction to The Netherlands in 2001 and show that the observations on the two veal-calf farms were not impossible.

Purpose of Review: Interventional procedures in the electrophysiology and catheterization laboratory are rapidly advancing. Critical to the advancement of these procedures is accurate identification of critical anatomic landmarks and catheter position. Fluoroscopy remains the mainstay for general identification of anatomic landmarks but is inadequate for the precise imaging needed for complex procedures. Precise imaging of anatomic landmarks and catheter position is now possible during the procedure with the use of intracardiac echocardiography (ICE). This paper reviews the rapid development and utilization of ICE in interventional electrophysiology., Recent Findings: Several recent studies show ICE as a major contribution to providing a safer, more reliable, and more cost-effective means of accomplishing the tasks performed by existing techniques. In the electrophysiology laboratory, the dependence on this new technology has been due to the rapid development of catheter-based radiofrequency ablation of the pulmonary veins for treatment of atrial fibrillation. Since the initial use of ICE in facilitating ablation of atrial fibrillation, other uses for ICE are continuously being identified., Summary: A comprehensive look is provided at the history and development of this new technology along with the most recent applications of ICE in interventional electrophysiology.

Objective: To determine the within-host population dynamics of Campylobacter jejuni and Escherichia coli in chickens during and after treatment with fluoroquinolones., Materials and Methods: Total and resistant faecal counts were determined from cloacal swabs during and after treatment with enrofloxacin. Chickens were housed individually to avoid confounding as a result of interaction between animals, and to be able to focus solely on the within-host dynamics. To determine the molecular basis of resistance, a number of isolates were checked for mutations in gyrA., Results: Treatment with enrofloxacin at doses routinely prescribed (50 ppm) rapidly reduced the faecal counts of E. coli below the detection limit and did not induce resistance. In C. jejuni, on the other hand, treatment with enrofloxacin quickly selected for high frequencies of fluoroquinolone-resistant strains. In all phenotypically resistant isolates, resistance was traced to mutations in the gyrA gene., Conclusions: (1) A licensed dosage (50 ppm) of enrofloxacin in drinking water of chickens is effective (i.e. markedly reduced faecal counts) and is safe on a short time scale in E. coli (i.e. did not rapidly select for resistance), but is neither safe nor effective in C. jejuni. (2) The rapid emergence of resistance to quinolones in C. jejuni does not necessarily result from horizontal transmission of resistant strains among chickens, but could solely be the result of de novo selection of resistance in individual chickens.

The objective of this study was to determine if calculation of the sinus of Valsalva to annulus ratio (SOV/Ann) effectively screens for root dilatation (ARD) during two-dimensional (2D) echocardiography in young patients (1 month-15 years) with suspected or confirmed Marfan syndrome. The aortic root was imaged using 2D echocardiography (parasternal long-axis view) and the SOV/Ann ratio calculated/analyzed in 27 Marfan patients and 45 age-matched controls. All controls had a SOV/Ann ratio <1.45, and 82% of Marfan's with ARD had a SOV/Ann ratio 1.45. Five of the Marfan patients with ARD had a SOV/Ann ratio < 1.45 due to normalization of the SOV/Ann ratio because of pathologic annular enlargement. Determination of the SOV/Ann ratio allows rapid on-line screening for aortic root dilatation during 2D echocardiography. A SOV/Ann ratio 1.45 predicted ARD in young Marfan patients with a high degree of accuracy (sensitivity = 0.82, specificity = 1.00). Screening for ARD using the SOV/Ann ratio fails when annular dilatation has progressed to the degree that the SOV/Ann ratio normalizes (SOV/Ann < 1.45). In this subgroup of patients, determination of aortic root dilatation will have to be evaluated by standard methods.

Isolation of the left innominate artery arising from the ductus arteriosus is a rare clinical entity with only nine cases being reported in the literature [5, 7, 8]. We describe a case consisting of tetralogy of Fallot, right aortic arch, and isolation of the left innominate artery arising from a left-sided ductus arteriosus which, to the best of our knowledge, has not been previously reported. The embryology of the aortic arch anomaly is reviewed.

We describe the first two successful cases of transcarotid balloon aortic valvuloplasty for critical aortic valve stenosis in the neonate utilizing continuous transesophageal echocardiographic guidance performed at the bedside. This method obviates the need for transporting a sick and hemodynamically unstable neonate to the catheterization laboratory and additionally reduces costs without compromising clinical care.

Neonatal critical aortic valve stenosis is a life-threatening malformation if untreated. Before the late 1980s, the preferred treatment was surgical valvotomy; however, operative mortality was high. Early reports of transcatheter balloon dilation were encouraging, although femoral artery damage and aortic valve insufficiency were procedural limitations. With new balloon catheter technology, transumbilical, transvenous, and transcarotid approaches have been advocated, although a comparison with recent surgical results has not been performed. We compared all neonates who presented to our institution since 1985 with the diagnosis of critical aortic stenosis. Ten patients underwent surgical transventricular valvotomy and 13 patients underwent balloon valvuloplasty via a right carotid cutdown with continuous transesophageal echocardiographic guidance. Prior to intervention, all patients had either left ventricular dysfunction, an aortic valve gradient > 100 mmHg, significant mitral valve insufficiency, and/or ductal dependent systemic blood flow. All patients had successful relief of aortic valve obstruction with normalization of left ventricular function and successful discontinuation of prostaglandin E1. Use of continuous transesophageal echocardiographic guidance resulted in fluoroscopic exposure of only 12 +/- 8 minutes. At the latest follow-up, a similar proportion of patients has required additional aortic valve procedures (38% vs 25%) and overall mortality (20% vs 15%) is similar. In the transcarotid group, 9 of 13 patients (69%) have a normal appearing right carotid artery by Duplex imaging, and no neurologic events have been reported. Balloon aortic valvuloplasty via a right transcarotid approach is safe, simplifies crossing the valve, and is effective for the initial palliation of neonatal critical aortic stenosis. The use of transesophageal echocardiographic guidance reduces fluoroscopy exposure, enables accurate assessment of hemodynamics without catheter manipulation or angiography, and avoids femoral artery injury.

Cytomegalovirus inclusions have been reported in perineal ulcers from immunosuppressed adults. The importance of this finding is unknown. We report the first pediatric case of cutaneous cytomegalovirus infection in an infant with congenital human immunodeficiency virus infection, presenting as a diaper dermatitis. Cytomegalovirus was cultured from the skin biopsy specimen, and characteristic inclusions were seen on hematoxylin-eosin-stained sections. Results of this biopsy specimen analysis prompted further investigation revealing disseminated cytomegalovirus infection, including retinitis. Aggressive pursuit of a pathogen in common conditions such as diaper dermatitis is strongly recommended in immunosuppressed pediatric patients.