C. van Maanen

Equine influenza virus infections: An update

Summary Equine influenza is one of the most economically important contagious respiratory diseases of horses. In this paper the current state of knowledge of equine influenza virus and the most important aspects of these virus infections, e.g. epidemiology, clinical aspects, pathogenesis and pathology, immunity, diagnosis, treatment, management and vaccination, are reviewed with an emphasis on epidemiology, diagnosis and vaccinology. Many questions have remained and with the advent of improved technology new questions have arisen. Consequently, research priorities should be set in an attempt to answer them. Therefore, this review ends with some personal recommendations for important priorities for future research.

Interference of maternal antibodies with the immune response of foals after vaccination against equine influenza.

The purpose of the study was twofold. First, using two groups of 22 foals each, we investigated the extent to which maternal antibodies interfere with the humoral response against equine influenza. The foals were born to mares that had been vaccinated twice yearly against influenza since 1982. Foals of group I were vaccinated three times at early ages (12, 16, and 32 weeks of age), and foals of group II were likewise vaccinated but a later ages (24, 28, and 44 weeks of age). After the first and second vaccinations, neither group showed an increase in antibodies that inhibit haemagglutination. Group II foals, however, had a significantly stronger antibody response against nucleoprotein after the second vaccination than the foals of group I. After the third vaccination, group II foals had a significantly stronger and longer lasting antibody response against haemagglutinin than the foals of group I. However, the antibody response to nucleoprotein was comparable in both groups. Second, the foals of group II were studied to determine the persistence of maternal antibodies directed against a common nucleoprotein and the haemagglutinin of two strains of equine influenza A virus. Biological half-lives of 39, 32, and 33 days were calculated for maternal antibodies directed against haemagglutinin of strains H7N7 Prague and H3N8 Miami, and against the nucleoprotein respectively. Maternal antibody titres at the time of vaccination were closely related to the degree of interference with the immune response. Because even small amounts of maternal antibodies interfered with the efficacy of vaccination, we conclude that foals born to mares vaccinated more than once yearly against influenza virus should not be vaccinated before 24 weeks of age.

Equine herpesvirus 1 and 4 infections: An update

Summary Equine herpesvirus 1 (EHV1) and equine herpesvirus 4 (EHV4) are important ubiquitous equine pathogens, causing much damage to the viral horse industry. EHV1 strains are associated with respiratory disease, abortion, and paresis/paralysis, whereas EHV4 strains are predominantly associated with respiratory disease. In the past decades much research effort has been put into improving knowledge about these viruses. In this paper the current state of knowledge of these viruses and the most important aspects of these virus infections, e.g. epidemiology, clinical aspects, pathogenesis and pathology, immunity, diagnosis, preventive management and management in the course of an outbreak and vaccination, is reviewed. Because we performed some research ourselves in the areas of diagnosis, epidemiology and vaccinology these aspects are reviewed in more depth than the other aspects. Still many questions have remained and new questions have risen. Consequently, research priorities should be made in an attempt t...Summary Equine herpesvirus 1 (EHV1) and equine herpesvirus 4 (EHV4) are important ubiquitous equine pathogens, causing much damage to the viral horse industry. EHV1 strains are associated with respiratory disease, abortion, and paresis/paralysis, whereas EHV4 strains are predominantly associated with respiratory disease. In the past decades much research effort has been put into improving knowledge about these viruses. In this paper the current state of knowledge of these viruses and the most important aspects of these virus infections, e.g. epidemiology, clinical aspects, pathogenesis and pathology, immunity, diagnosis, preventive management and management in the course of an outbreak and vaccination, is reviewed. Because we performed some research ourselves in the areas of diagnosis, epidemiology and vaccinology these aspects are reviewed in more depth than the other aspects. Still many questions have remained and new questions have risen. Consequently, research priorities should be made in an attempt to answer these questions. Therefore, this review ends with some personal recommendations for important priorities for future research.

Airborne transmission of bovine herpesvirus 1 infections in calves under field conditions

A small scale transmission experiment was performed with bovine herpesvirus 1 (BHV1) in a cattle population under field conditions. 10 calves were housed under strict hygienic conditions, with a distance of 4m between each calf. Five calves were experimentally infected with BHV1, two calves with strain Harberink and three with strain Lam, respectively. Experimentally infected calves were placed at 4 m distance from five susceptible sentinel calves. Airborne transmission to sentinel calves was detected using virus isolation and BHV1 specific polymerase chain reactions in samples of nasal fluids, and BHV1 specific antibodies in serum samples. Strain Harberink was hardly transmitted to sentinel calves, whereas strain Lam was transmitted to all sentinels. Estimating the rate of transmission per day, the total number of calves infected by one (strain Lam) infected calf was 1.18. Comparing this estimated transmission ratio between cattle at a distance of 4 m to the estimated transmission ratio R of BHV1 in susceptible commingled cattle reported before, the effect of the factor distance on the transmission ratio could be calculated. Extrapolating these results, a distance of 4.4 m between cattle populations would be necessary to reduce transmission for this strain to R<1.

Diagnostic performance of ELISA and PCR in identifying SRLV-infected sheep and goats using serum, plasma and milk samples and in early detection of infection in dairy flocks through bulk milk testing.

The objective of the study was to evaluate the diagnostic performances of the ELITEST-MVV ELISA for detection of antibodies against small ruminant lentiviruses and of two recently published PCRs for the detection of proviral DNA of SRLV in blood and corresponding individual milk samples. In addition, the feasibility of bulk milk testing was investigated by titrating ELISA positive pooled milk samples in negative milk, and by investigating bulk milk samples by ELISA and PCR in relation to the SRLV-status of the flocks. The results show that plasma and milk are suitable replacements for serum. For sheep, both PCRs showed a better diagnostic performance than for goats. ELISA results for bulk milk samples were promising with a putative detection limit of <3% within-herd prevalence using 1/10 pre-diluted samples and even <1% within-herd prevalence when samples were tested undiluted. In a panel of 249 bulk milk samples, all samples from SRLV free flocks (n=138) tested negative in the ELISA, while 50% of the samples from flocks with an unknown SRLV-status (n=111) were positive. For a subset of 59 bulk milk samples, agreement between ELISA results and leader-gag PCR results was almost 100%. These results demonstrate the potential of bulk milk testing as a cost effective tool for early detection of infection in dairy flocks, which is essential for SRLV-monitoring programs.

Comparison of three polymerase chain reaction methods for routine detection of bovine herpesvirus 1 DNA in fresh bull semen

Five bulls were inoculated intrapreputially with Bovineherpes virus 1 (BHV 1), in order to compare the relative sensitivity of three polymerase chain reaction (PCR) assays for routine diagnosis of fresh bovine semen for the presence of BHV 1 Semen was collected twice a week up to 107 days post-infection (dpi). To reactivate latent virus, the bulls were treated with dexamethasone from 44 until 48 dpi. All samples were examined before and after cryopreservation treatment using a standard virus isolation (VI) method and three PCR assays: PCR A, PCR B and PCR C. PCR A and PCR C used an internal control plasmid DNA template and PCR B used the split sample method in order to control for false negative results. Of the 149 fresh semen samples that were tested, PCR A detected 45 positive, PCR B detected 39 positive and PCR C detected 66 positive, while virus was isolated from 22 samples. Of the 149 samples treated by cryopreservation, the virus was isolated from 13 samples and PCR C was positive in 21 samples. The results demonstrate that all three PCR assays are more sensitive than virus isolation, particularly during the later phases of infection.

Neurological syndromes among horses in The Netherlands. A 5 year retrospective survey (1999-2004).

Summary The presence of toxins or infectious agents combined with environmental factors in combination susceptible host can be the with a cause for neurological disease in groups of horses. During a 5 year observational period outbreaks of neurological diseases among horses were evaluated. Causes of occurring neurological diseases were equine botulism, lolitrem intoxications, equine herpesvirus type 1‐associated myelo(encephalo)pathy, and encephalitis caused by (disseminated) Streptococcus equi subspecies equi infection. This article focuses on the first three syndromes because of their predominant influence on locomotion. The pathogenesis of each disease is presented, followed by a description of a general presentation of the diseases as encountered under Dutch circumstances.

An equine herpesvirus 1 (EHV1) abortion storm at a riding school

Summary An outbreak of EHV1 abortions occurred at a riding school in the Netherlands in 1991. Seven of twelve pregnant mares aborted, and another foal died at 8 days of age. Six abortions occurred within 12 days in March after an initial abortion on 8 February. Four mares delivered live foals. Virological examination of four aborted foals revealed an EHV1 infection. Serological results for paired sera from 17 horses suggested, that the initial abortion on 8 February was the index case, and probably caused the other six abortions. The index case could well have been caused by reactivation of latent virus induced by transport stress. The laboratory results are discussed in the light of the present knowledge of the pathogenesis and epidemiology of EHV1 abortion.

Schmallenberg virus antibodies in bovine and ovine foetuses

Since December 2011, an epizootic of congenital malformations, designated as arthrogryposis-hydranencephaly syndrome (AHS), has been seen in ruminants in several countries in Western Europe (Van den Brom and others 2012). The disease has been associated with a new orthobunyavirus, named Schmallenberg virus (SBV) (Hoffmann and others 2012). After SBV was detected in brain tissue from malformed lambs by PCR on December 15, 2011, the Dutch Ministry of Economic Affairs, Agriculture and Innovation made congenital malformations in ruminants notifiable. Farmers and veterinarians are obliged to notify and submit malformed newborns to the Animal Health Service (GD-Deventer) for autopsy examination. Until May 15, 2012, over 600 lambs and over 1200 calves had been examined, and many of them showed AHS which is likely to be caused by an intrauterine infection with SBV. Brain tissue samples of all malformed newborns were tested by PCR at the Central Veterinary Institute (CVI). However, SBV could only be confirmed in 22 per cent of the lambs and 19 per cent of the calves that were investigated until May 15, 2012. Initially, a real-time PCR protocol targeting the L-segment was used. Since February 17, 2012, a real-time PCR protocol targeting the S-segment was used, since the latter PCR demonstrated a higher analytical and diagnostic sensitivity as indicated by the Friedrich Loeffler Institute in Germany and Dijkman and others (2012). Both protocols were kindly provided by the Friedrich Loeffler Institute in Germany. Persistent infection in Akabane virus-infected foetuses has not been reported, and the virus cannot be easily isolated from affected full-term calves or lambs because of the presence of neutralising (NT) …

Experimental infection with neuropathogenic equid herpesvirus type 1 (EHV-1) in adult horses.

Equid herpesvirus type 1 (EHV-1)-associated myeloencephalopathy (EHM) may follow an infection with the virus in horses. This study tested three hypotheses: (1) a large inhaled dose of a neuropathogenic EHV-1 strain would induce a cell-associated viraemia in all infected horses; (2) neurological disease will only occur in viraemic horses, and (3) the cerebrospinal fluid (CSF) composition following EHV-1 viraemia will be an indicator for EHM. Four EHV-1 seronegative horses were inoculated with EHV-1 by inhalation. Three developed clinical signs of neurological disease, which were mild in two horses and lacking typical EHM histopathological findings, but moderately severe in the third horse. This latter animal was the only one found to be viraemic, with xanthochromic CSF and spinal cord histopathology findings characteristic of EHM. This study showed that cell-associated viraemia was not guaranteed, despite a large-dose inoculation with EHV-1, yet viraemia was probably a pre-requisite for subsequent development of EHM. The histopathological changes used to confirm EHM may be predicted from CSF analysis.