Biomedical subjects

F Koenen

Publications and source records attributed to F Koenen.

At least 37 records · Page 2Linked to original sources

PubMed2001

Descriptive epidemiology of a classical swine fever outbreak in the Limburg Province of Belgium in 1997.

This paper describes the epidemiological characteristics of the 1997 Classical Swine Fever (CSF) outbreak that occurred in the Limburg Province of Belgium, where there is a policy of non-vaccination, intensive surveillance and eradication. Between 30 June and 17 July 1997, eight herds, located in three different areas, were confirmed to be CSF-positive. CSF virus was transmitted from the primary infected herd of one area to another five herds in the same area and to one herd in a different area. The mode of virus introduction for this primary infected herd and for the one herd that was not infected by this primary herd could not be determined. Clinical, serological, and virological findings indicated that the CSF-infected herds were detected in an early stage of the infection. The early detection of the infection together with a preventive stamping out procedure resulted in a rapid elimination of the CSF virus. A total of 46,561 pigs were slaughtered to control the spread of the infection. Another 27,579 pigs were slaughtered in the framework of the market support. The total direct costs of the episode were estimated at [symbol: see text] 10,893,337.

Animals↗

PubMed2001

An experimental infection with classical swine fever virus in pregnant sows: transmission of the virus, course of the disease, antibody response and effect on gestation.

An experimental infection with classical swine fever (CSF) virus in 12 conventional gilts, housed in a sow-box housing system, was conducted in order to evaluate horizontal transmission, clinical, virological and serological response, and the effect on gestation. Two of the 12 gilts, of which 10 were pregnant, were experimentally inoculated. They became viraemic for the first time 6 days post-inoculation (dpi). The contact gilts became viraemic between 18 and 21 days post inoculation. On the basis of virological findings and the martingale estimate of R0 (13.0) it was concluded that the two experimentally inoculated gilts infected all contact gilts, although random contacts between gilts were not possible. The prescence of CSF infection could be diagnosed earlier and during a longer period when the leucocyte count or polymerase chain reaction were used in comparison with virus isolation in whole blood (P < 0.05). The observed clinical symptoms were atypical and highly variable between the gilts, which hampered clinical diagnosis. The pregnant gilts became infected between day 43 and 67 of gestation. In all cases vertical virus transmission occurred and this resulted partially in abortion and/or mummification.

Abortion, Veterinary↗

PubMed2000

Molecular epidemiology of a large classical swine fever epidemic in the European Union in 1997-1998.

A big epidemic of classical swine fever (CSF) occurred in the European Community in 1997. The first case was reported at the beginning of January 1997 from Germany. The disease presumably spread to the Netherlands, and from there to Italy, Spain and eventually to Belgium. About 30 isolates from these outbreaks were analysed by comparison of the nucleotide sequence data generated from fragments of both the E2 glycoprotein gene (190 nucleotides) and from the 5'-nontranslated region (5'-NTR; 150 nucleotides). By combining epidemiological data with genetic typing, it was found that the outbreaks were related and caused by a virus belonging to the genetic subgroup 2.1. As this type of virus had been reported infrequently in Europe and not at all since 1993, we postulate that it was newly introduced into the European Union (EU).

5' Untranslated Regions↗

PubMed2000

Classical swine fever virus: a second ring test to evaluate RT-PCR detection methods.

Six laboratories participated in a study to compare the sensitivity and specificity of RT-PCR tests for the detection of classical swine fever virus (CSFV). Sets of coded samples were prepared by serial dilution of positive samples and then distributed to each of the laboratories. One set comprised 25 samples of random primed cDNA, synthesised from viral RNA representative of different pestiviruses. The other set comprised samples of blood and serum obtained from virus-free or CSFV-infected pigs. Each laboratory tested the samples using PCR/RT-PCR according to a set of standardised protocols that specified the exact conditions and requirements for inclusion of control samples. Two types of test were evaluated. One amplified a part of the 5'-non coding region of the pestivirus genome by means of a closed, one-tube RT-nested PCR. The other amplified a part of the NS5B gene using non-nested RT-PCR. The results of the laboratories were compared with one another, and with those obtained earlier when similar samples were tested by the same laboratories using non-standardised methods [Paton et al., Classical swine fever virus: a ring test to evaluate RT-PCR detection methods, Vet. Microbiol., in press]. Standardisation of the protocols resulted in a more consistent test sensitivity. Three laboratories avoided significant false positive results. Others that did not, could nevertheless recognise that test specificity was inadequate from the results obtained with the control samples. Minimum requirements for the inclusion of adequate controls and periodic proficiency testing are proposed.

Animals↗

PubMed2000

An experimental infection with classical swine fever in E2 sub-unit marker-vaccine vaccinated and in non-vaccinated pigs.

The clinical and virological protection induced by an E2 sub-unit marker-vaccine against Classical Swine Fever (CSF) was examined during an experimental infection in vaccinated and non-vaccinated pigs. Forty-five pigs were equally distributed over three adjacent pens of an isolation unit, there was only indirect (airborne) contact between pigs in the different pens. In pen 3 all pigs were vaccinated twice with 4 weeks interval. Pigs in pens 1 and 2 were not vaccinated. Two weeks after booster vaccination, one randomly selected pig in the middle pen was experimentally inoculated with CSF virus. After the initial virus spread in the infected pen, all pigs in the non-vaccinated adjacent pen were infected. In the vaccinated pen, seven out of 14 pigs became infected during the experiment. Survival analysis showed that virus transmission by direct and indirect contact was significantly (p<0.001) delayed in vaccinated pigs as compared to non-vaccinated pigs. In the non-vaccinated pens over 40% of the pigs died and typical clinical signs were noticed. In the vaccinated pen no mortality and no clinical symptoms were observed. Although double vaccination with an E2 sub-unit marker-vaccine was able to prevent the clinical course of the disease it was unable to prevent infection through indirect contact. This finding combined with the slow serological response after vaccination will complicate the possible use of the vaccine in emergency vaccination programmes.

Animals↗

PubMed2000

Classical swine fever virus: a ring test to evaluate RT-PCR detection methods.

Six laboratories participated in an exercise to compare the sensitivity and specificity of RT-PCR tests for the detection of classical swine fever virus (CSFV). Two sets of coded samples were prepared by serial dilution of positive samples and then distributed to each of the laboratories. One set comprised 34 samples of random primed cDNA. These had been synthesised from viral RNA representative of seven different genetic subtypes of CSFV. The other set comprised 40 clinical samples containing tonsil, spleen, whole blood or serum from a pig that had been experimentally infected with CSFV. Each laboratory tested the samples using one or more PCR/RT-PCR tests that they were accustomed to using. The methods and results of the laboratories were compared with one another. The RT-PCR results obtained from testing the clinical samples were also compared with those obtained by virus isolation and antigen ELISA.ELISA. Both RT-PCR and RT-nested PCR appeared to give some false positive results. Several of the PCR tests appear suitable in terms of specificity and sensitivity. Further trials are necessary to compare results when the same test is performed by different laboratories, and to show that improved control procedures can eliminate problems due to false positive reactions.A limited comparison of extraction and reverse transcription procedures showed similar results in each of three participating laboratories, even though the methods were not standardised.

Classical Swine Fever Virus↗

PubMed1999

Experimental infection of slaughter pigs with classical swine fever virus: transmission of the virus, course of the disease and antibody response.

The spread of classical swine fever virus was investigated in an isolation unit containing four pens, each containing six slaughter pigs. One pig in the middle pen of three adjacent pens was inoculated intramuscularly and intranasally with the virus. The fourth pen was located in a separate compartment. The pens were visited in a strict order to study, first, the effect of indirect contact via contaminated clothing and footwear on the spread of the virus to adjacent pens and, secondly, the airborne transmission of the virus between compartments. The pigs were examined and blood samples were taken every other day for 62 days for virological and serological analyses. The virus was highly contagious for the five pigs that were in direct contact with the inoculated pig, but spread to the other pens only after all the pigs in the originally infected pen had become viraemic. The spread of the virus was promoted by contaminated clothing and footwear, but airborne transmission contributed considerably to the spread of the virus within the pighouse. The first clinical signs observed after the virus was introduced into a pen were decreased feed intake, increased mean rectal temperature and apathy. Neither the clinical course of the infection, nor the pattern of seroconversion observed over time, was affected by the differences in the intensity of contact with the virus between the pigs in the different pens.

Animals↗

PubMed1999

Prevalence of herds with young sows seropositive to pseudorabies (Aujeszky's disease) in northern Belgium.

In Belgium, pseudorabies in swine has been the subject of a mandatory eradication programme since 1993. From December 1995 to February 1996, a survey was conducted in the five provinces of northern Belgium to estimate the provincial pseudorabies virus (PRV) herd seroprevalence. Seven hundred and twenty randomly selected herds were included in this survey. To detect recently infected animals, only young sows were sampled. The results show that 44% of these herds had an important number of PRV-seropositive young sows. The highest herd seroprevalence was observed in West Flanders (68%), followed by Antwerp (60%), East Flanders (43%), Limburg (18%), and Flemish Brabant (8%). Assuming a diagnostic test sensitivity and specificity of 95% and 99%, respectively, and a true PRV within-herd prevalence of 43%, the overall true PRV herd prevalence was estimated to be 35%. A logistic multiple-regression revealed that the presence of finishing pigs was associated with a two-fold increase in odds of a herd being seropositive (odds ratio (OR)=2.07, 95% confidence interval (CI) = 1.31-3.26); a breeding herd size > or =70 sows was associated with a four-fold increase in odds of a herd being seropositive (OR = 4.09, 95% CI = 2.18-7.67); a pig density in the municipality of >455 pigs/km2 was associated with a 10-fold increase in odds of a herd being seropositive (OR = 9.68, 95% CI = 5.17-18.12). No association was detected between the PRV herd seroprevalence and purchase policy of breeding pigs (purchased gilts, or use of homebred gilts only).

Animal Husbandry↗

PubMed1999

Phylogenetic analysis of European encephalomyocarditis viruses: comparison of two genomic regions.

The phylogenetic relationships of encephalomyocarditis (EMC) viruses isolated from pigs and rodents in Europe were determined by comparison of nucleotide sequences from two different regions of the virus genome, the VP3/VP1 gene junction (part of the capsid-coding region) and part of the 3D polymerase-coding region. Thirty-five European EMC viruses could be divided into two genetic groups, one which contained viruses from Greece isolated between 1986 and 1997 and from Belgium in 1991 and the other which contained viruses from Italy (1986-1996), Cyprus (1994-1995), France (1995) and Belgium (1995-1996).

Animals↗

PubMed1999

Classical swine fever virus is genetically stable in vitro and in vivo.

Phylogenetic analyses of large numbers of classical swine fever strains have revealed a high degree of sequence conservation in the genomic regions examined, suggesting either a recent common ancestor or a low evolution rate. This low variability is in contrast to findings with other RNA viruses. To investigate the consequence of this apparent genetic stability on phylogenetic examinations, the Belgian field isolate Wingene'93 was passaged in pigs as well as in cell culture by various methods. Sequence analyses of viruses collected after various passages in three target regions proposed for phylogenetic studies (5' NTR, E2, and NS5B) revealed a complete sequence conservation. Only when the amount of passaged virus was lowered, mimicking a genetic bottleneck, a single point mutation was observed in the E2 gene. Additionally, only four nucleotide substitutions were observed when the genome of a virus obtained after 96 cell passages in persistently infected cells was compared with its parental virus, the recombinant virus derived from an infectious cDNA clone of CSFV strain Alfort/187. This low mutation frequency observed both in vitro and in vivo demonstrates that classical swine fever virus is genetically stable. Hence, even minor mutations can be considered significant in molecular epidemiological studies.

Animals↗

PubMed1999

Epidemiologic, pathogenic and molecular analysis of recent encephalomyocarditis outbreaks in Belgium.

In 1991 EMCV was isolated for the first time in Belgium from the offspring of a sow with reproductive failure. From August 1995 until December 1996, EMCV was diagnosed in 154 Belgian pig holdings in association with myocardial failure and sudden death in fatteners and suckling piglets or with reproductive failure in sows. To clarify some epidemiological aspects 3 EMCV isolates characteristic for the different clinical pictures and outbreaks were studied. Field observations and animal experiments indicated that the pathogenicity induced by each isolate is specific for one age category and that the spread of the virus is limited. The presented data also suggest that rodents may play a role in the transmission of EMCV but that pig-to-pig transmission is at least as important. Molecular analysis of two separate regions on the genomes of the respective EMCV isolates showed that the 1995-96 EMCV epizootic in Belgium was due to a new virus introduction. Furthermore, the VP1 coding gene is proposed as a marker of virulence.

Amino Acid Sequence↗

PubMed1998

An experimental infection with classical swine fever virus in weaner pigs. I. Transmission of the virus, course of the disease, and antibody response.

The spread of Classical Swine Fever (CSF) virus (strain Lorraine), originally isolated in the first CSF infected herd of the 1993-1994 Belgian epizootic, was examined in an isolation unit with three adjacent pens and 15 weaner pigs per pen. Virus was introduced through experimental inoculation of one weaner pig in the middle pen (pen 2). The experimentally inoculated pig became viraemic 4 days post-inoculation (dpi) and the pen mates at 12 (n = 9) and 14 dpi (n = 5). The first viraemia in pens 1 and 3 was observed 18 dpi. Pigs were found to be seropositive in pens 1, 2, and 3 from 24, 20, and 22 dpi onwards, respectively. The reproduction ratio (R0) for the pigs in pen 2, estimated according to the martingale method, was 81.3 (s.e. = 109.54). The rate ratio (Cox proportional hazard) of the first pigs to become viraemic in pen 3 (airborne contact plus contact via contaminated clothing and footwear with pen 2) versus pen 1 (airborne contact with pen 2 only) was 1.60 (P = 0.3342). Thus, the additional contact of contaminated clothing did not affect transmission of the CSF virus. The survivor function (Kaplan-Meier survival analysis) did not significantly differ per pen. The time from first detection of virus in plasma to death was not significantly different between pens. The mean rectal temperature of pigs in a pen increased 3 to 4 days prior to detection of virus. The proportion of seropositive pigs per pen (p) from the day the first weaner pig in a pen became viraemic (dpf) was examined as a function of time with a logistic regression model. The model parameter estimates did not differ between pens. Hence, the data from the three pens were pooled. The regression equation of the seroprevalence over time for the pooled data was p = 1/[1+e(4.65-0.39 *dpf)].

Animal Husbandry↗

PubMed1998

An experimental infection with a classical swine fever virus in weaner pigs. II. The use of serological data to estimate the day of virus introduction in natural outbreaks.

In a companion paper a logistic regression model of seroprevalence over time was developed on the basis of data obtained during an experimental infection of weaner pigs with classical swine fever (CSF) virus. The model was applied to seroprevalence data from three outbreaks of the 1993-1994 epizootic to test whether the model could predict correctly the day of virus introduction into the herd. It was concluded that the logistic regression model has potential as a tool to estimate in retrospect the day CSF virus was introduced into a pig herd, which in turn may assist in identification of risk factors implicated in the further spread of the disease.

Animals↗

PubMed1998

Identification of encephalomyocarditis virus in clinical samples by reverse transcription-PCR followed by genetic typing using sequence analysis.

The objective of the present study was to gain a better understanding of the epidemiology of encephalomyocarditis virus (EMCV) infections in pigs by applying molecular techniques. The diagnostic potential of a reverse transcription-PCR (RT-PCR) targeting 286 nucleotides at the 3' end of the gene which encodes the viral polymerase was assessed with experimental and field samples. In addition, the use of the amplified sequences for an epidemiological study was evaluated. The heart was clearly shown to be the most suitable organ. The detection limit was determined to be 1 viral particle in 100 mg of heart tissue. The sensitivity and specificity of the assay on the basis of the results obtained in this study were 94 and 100%, respectively. Phylogenetic analysis of the amplified sequences classified EMCVs in two distinct lineages. Group A consists of the reference strain ATCC 129B, all isolates collected between 1991 and 1994 in Belgium in association with reproductive failure, and all Greek isolates. All Belgian isolates collected since the first isolation of EMCV in relation to myocardial failure in fatteners in Belgium group together with the isolates from Cyprus (1996 and 1997), Italy (1986 to 1996), and France (1995) in group B irrespective of their pathogenicity. The analyzed part of the 3D gene differed by 13.0% between Groups A and B. In contrast to the sequence homogeneity of the Belgian isolates collected between 1991 and 1994, molecular diversity, which ranged between 0 and 2%, was observed among the Belgian isolates collected in 1995 and 1996. Among all Greek isolates the diversity ranged between 1 and 8%. However, this diversity does not seem to reflect geographical links between the outbreaks. A RT-PCR for the rapid and specific diagnosis of EMCV in a variety of clinical samples followed by nucleotide sequence analysis proved to be valuable for molecular epidemiological studies.

Animals↗

PubMed1997

Rapid diagnosis of encephalomyocarditis virus infections in pigs using a reverse transcription-polymerase chain reaction.

Encephalomyocarditis virus (EMCV) is widespread and the economic losses caused by an EMCV outbreak in pig holdings and the similarity between a foot-and-mouth disease virus (FMDV) and an EMCV infection in young piglets stress the need for a rapid, specific and broad diagnostic assay. An alternative to the time-consuming seroneutralisation assay, currently used for the characterisation of EMCV, is described. An EMCV specific reverse transcription-polymerase chain reaction (RT-PCR), using primers located in a conserved region of the 3D gene of the viral genome, was developed and tested on 114 different EMCV isolates. The identity of the respective amplicons was confirmed by sequencing. The potential of this assay for future diagnostic purposes was demonstrated by applying the RT-PCR on tissue samples collected from an experimentally infected piglet.

Animals↗

PubMed1997

Comparative study of the pathogenic properties of a Belgian and a Greek encephalomyocarditis virus (EMCV) isolate for sows in gestation.

Thirteen conventional sows were inoculated between 61 and 92 days of gestation with a Belgium or a Greek EMCV isolate to investigate the difference in pathogenicity of both strains for sows in gestation. The Belgian EMCV strain was isolated from the offspring of a sow with productive failure and without myocardial lesions. The Greek strain was isolated from a 3-month-old pig with prominent myocardial lesions. The present study demonstrates a transplacental virus transmission with fetal death following an infection of the sows in gestation with both isolates. The fetal pathogenicity was more severe with the Greek strain than with the Belgian isolate. No myocardial lesions were noticed.

Animals↗

PubMed1996

Epidemiological characteristics of an outbreak of classical swine fever in an area of high pig density.

The objective of this study was to analyse an outbreak of classical swine fever under a policy of non-vaccination, intensive surveillance and eradication in an area of high pig density. The virus was found in 52 herds, where some 90,000 pigs were slaughtered. The clinical signs were vague and the reports of suspect herds generally coincided with increased mortality. The interval between the first occurrence of clinical signs and the report of a suspect herd was shorter when the disease was first diagnosed in fattening pigs than when it was diagnosed in sows, boars or suckling piglets. Among fattening pigs, mortality and morbidity appeared to increase with age. The proportion of clinically ill animals was positively correlated with the proportion of serologically positive animals in a pig house during the phase when the disease was spreading. Fifty-eight per cent of pig houses containing only clinically healthy but some virologically positive pigs were serologically negative. Antigen detection was therefore critical for early disease detection. Serology was nevertheless useful to ascertain that swine fever was not endemic in the area. The secondary cases were concentrated in the close neighbourhood of the herd initially infected.

Animals↗

PubMed1995

Evaluation of the enzyme-linked immunosorbent assay for the rapid screening and detection of classical swine fever virus antigens in the blood of pigs.

A workshop was convened, at which seven enzyme-linked immunosorbent assays (ELISAs) were compared with virus isolation for the detection of viraemia in serial blood samples collected from six pigs at up to fourteen days after inoculation with classical swine fever virus. All ELISAs were of the double antibody sandwich type, using monoclonal and/or polyclonal antibodies to detect a variety of viral proteins in leukocytes, or in anti-coagulated blood or serum. Compared to virus isolation, specificity of the ELISA was good: only one sample found negative by virus isolation yielded a positive result in a single ELISA. Some false-negative results occurred with samples collected at up to eight days after inoculation, but all tests found samples collected between nine and fourteen days post-inoculation to be positive. The ELISAs require less-specialised facilities and can be performed much more rapidly than virus isolation. They are therefore extremely promising tools for screening large numbers of live pigs.

African Swine Fever↗