Antibodies against bovine herpesvirus (BHV) 5 may be differentiated from antibodies against BHV1 in a BHV1 glycoprotein E blocking ELISA

We examined whether antibodies against bovine herpesvirus (BHV) 5 cross-react with BHV1 antigens and whether they could interfere with BHV1 eradication programmes. Six calves were experimentally infected with different doses of BHV5 strain N569; homologous antibodies were first detectable on day 11 post infection; they cross-reacted in a BHV1 virus neutralisation test, in a BHV1-glycoprotein (g)-B blocking ELISA and in a BHV1-gE ELISA, but not in a BHV1-gE blocking ELISA. This study indicates that, in ongoing BHV1 eradication programmes, based on vaccines that lack gE, BHV5 infections may not lead to false-positive serological reactions in case cattle are tested for BHV1-gE antibodies by the BHV1-gE blocking ELISA; antibodies against BHV5 may be differentiated from antibodies against BHV1. The BHV1-gE blocking ELISA may, therefore, offer opportunities for the serological differentiation between BHV1 and BHV5 infections.     

Seroepizootiologic studies of the distribution of bovine herpesvirus type 1 and 2 (BHV 1, BHV 2) in Namibia

From cattle breeding areas of Namibia a total of 2,800 sera from 1984-1987 were tested by Elisa for antibodies to BHV1 and BHV2. From north-east to south-west a drop in BHV2 antibody prevalences was found which paralleled the decrease in rainfall. With BHV1 the percentage of positive reacting sera was also highest in the northern communal areas (up to 80%). In the commercial farming area with best farming conditions the percentage was lowest, but it was south of here again higher in the areas of lower precipitation where farming conditions are less optimal.     

Dynamics of infection and immunity in a dairy cattle population undergoing an eradication programme for Infectious Bovine Rhinotracheitis (IBR)

Several countries within the European Union (EU) have successfully eradicated Infectious Bovine Rhinotracheitis (IBR), while others (e.g. Germany) are making efforts to achieve IBR-free status. EU member states IBR eradication programmes must meet Community legislation requirements that ban breeding farms from purchasing positive animals, from using whole-virus IBR vaccines, and from inseminating cows with semen from positive bulls. A follow-up study from 2002 to 2005 was carried out in the province of Trento (Italy), where a compulsory programme for IBR eradication was started in 1998. IBR outbreaks (identified on the basis of seroconversion of sentinel animals) were concentrated in larger positive herds. A higher incidence was recorded between 2003 and 2004. An association between markedly high temperatures in the summer of 2003 and virus reactivation has been suggested but is yet to be confirmed. The practice of driving cattle to common alpine pastures for the summer season did not play a significant epidemiological role in IBR transmission. Premising that only seronegative animals are allowed to enter dairy farms, animal movement increases the infection risk to a moderate extent. The long-term persistence of IBR antibodies was more pronounced in animals positive for antibodies to the glycoprotein E (gE). Scattered seroconversions, occurring mostly in positive herds, require careful interpretation in order to avoid overestimating the incidence of the infection at herd level.     

Bovine herpesvirus type 4: a special herpesvirus (review article)

This paper summarizes the history of and information on bovine herpesvirus type 4 (BoHV-4) from the first isolation to the most recent results. For almost twenty years BoHV-4 has been considered a typical herpes 'orphan' virus, which infects several species but causes no illness. The latest experiments revealed the close relationship of this virus with the immune system and other tissues. The virus was even considered as a possible candidate for a vector vaccine. BoHV-4 as a strange herpesvirus has several features which are not characteristic of other herpesviruses, such as several latency sites, persistence in serum, dividing cells necessary for virus replication, and the wide host range. In addition to describing the main features of the virion, replication, clinical signs, nomenclature problems, this review intends to concentrate on the new and strange results coming out from several laboratories worldwide. It is also suggested that because the virus combines several properties of various herpesvirus subfamilies and because of its close relationship with the immune system, it may deserve further attention as a representative of a potentially new genus within the Gammaherpesvirinae subfamily.     

Bovine herpesvirus type 1 abortions detected by a semi-nested PCR in Brazilian cattle herds

Bovine herpesvirus type 1 (BHV-1) was investigated by a semi-nested polymerase chain reaction (SN-PCR) and by MDBK cell culture virus isolation in organ fragments from 55 aborted fetuses collected from beef and dairy cattle herds with history of reproductive problems in the North of Paraná State, Brazil. A 425 bp amplicon of the BHV-1 glycoprotein D gene was detected in 14 (25.4%) aborted fetuses. BHV-1 was isolated in MDBK cells from the tissue of 5 (9.1%) fetuses. The specificity of positive results was evaluated by Restriction Fragment Length Polymorphism (RFLP) with Bgl I restriction of DNA amplified by SN-PCR, and by virus-neutralization and immunofluorescence with rabbit anti-BHV-1 polyclonal antibodies for virus isolated in cell culture. The results of this work demonstrate the importance of using other diagnostic techniques, like SN-PCR, for BHV-1 detection in organ fragments from aborted fetuses and the high frequency of this virus in reproductive failures in Brazilian cattle herds.     

A polymerase chain reaction (PCR) assay for the detection of bovine herpesvirus 1 (BHV1) in selectively digested whole bovine semen

A PCR assay for the detection of bovine herpesvirus type 1 (BHV1) DNA in selectively digested whole bovine semen was developed and evaluated. A brief treatment with proteinase-K was used to lyse free virus, virus present in non-sperm cells and virus adhering to the spermatozoa. Genomic bovine DNA was not released by this treatment. Primers and probes were based on the nucleotide sequence of the gD gene. BHV1 virus-spiked split samples were used as positive controls and the PCR products were detected by eye in ethidium bromide-stained agarose gels. Sequentially collected non-extended semen samples from experimentally infected bulls were used to compare this assay with virus isolation. Of a total of 162 ejaculates, 51 were found positive by virus isolation, whereas PCR detected BHV1 DNA in 73. PCR detected BHV1 DNA for a longer period after infection and reaction. Apart from its superior sensitivity, this PCR assay also has the advantage of being a relatively simple procedure, providing results within 24 h.Abbreviations AI artificial insemination - BHV1 bovine herpesvirus type 1 - PCR polymerase chain reaction - TCID₅₀ tissue culture infective dose, 50%     

Bovine viral diarrhoea (BVD) infections--control and eradication programme in breeding herds in Slovenia

A Slovenian BVD control and eradication programme was initiated in 1994, and the results from testing of bovine herds for antigen and antibodies in 1996 are presented. Samples originating from breeding herds, breeding herds for young bulls, and insemination stations were tested by antigen or antibody ELISA, or by PCR. Out of 7968 samples from 354 herds we found 18% of the animals antibody-positive. In one region situated in the north-east of Slovenia we found the herds to be almost nearly free of BVDV infections (5% prevalence). No positive antigen ELISA findings were done in 374 blood samples from recruitment herds for young bulls, whereas two out of 206 sera were investigated by PCR-reacted positive. The differences in seroprevalence found between regions is thought to be caused by differences in summer pasturing and husbandry practices.     

Production objectives and breeding goals of Sahiwal cattle keepers in Kenya and implications for a breeding programme

The Sahiwal breed has been used for upgrading the East African Zebu (EAZ) for improved milk production and growth performance in the southern rangelands of Kenya. Main users of this breed are Maasai pastoralists. Until now, there has been no deliberate effort to understand why these pastoralists specifically prefer to keep Sahiwal genetic resources as well as which traits are considered important by them and what is the underlying reason for this. However, this information is regarded vital for further development of the breed. A survey was conducted between May and October 2009 among Maasai pastoralists in Kajiado and Narok counties in the Southern part of Kenya, and private ranches and government farms to identify production objectives and breeding goals of Sahiwal cattle producers. Sahiwal genetic resources were mainly kept for domestic milk production and for revenue generation through milk sales and live animals. To a limited extent, they were kept for breeding and also for multiple objectives that included insurance against risks and social functions. Production aims were influenced to varying extents by various household and farmer characteristics. Sahiwal cattle and their crosses were generally perceived to be better with respect to productive traits and fertility traits when compared to the EAZ. However, the EAZ was rated higher with respect to adaptation traits. The breeding objective traits of primary importance were high milk yield and big body size, good reproductive efficiency and relatively good adaptation to local production conditions. Performance and functional traits are important breeding goals that play a major role in fulfilling the multiple production objectives. This forms the basis for the optimisation of a breeding programme for sustainable utilisation to meet the needs of Sahiwal cattle producers.     

The detection of bovine herpesvirus type 1 (BHV 1) using an intradermal test. I. Field studies

An intradermal test (delayed hypersensitivity test) for the diagnosis of BHV1 infection was evaluated in 791 cattle of 16 dairy farms. The skin reactions were compared with the results of serological examinations using a commercial BHV1 ELISA kit (Trachitest). As antigen concentrated, purified and inactivated BHV1 was used. The skin reaction (increase of the skin fold thickness) was used for the interpretation of test results. The best results were obtained with the control of the skin reaction on the third day after injection of the antigen. From 393 serologically BHV1 negative cattle with an age of more than 6 months 391 (99.5%) had a skin reaction up to 1.0 mm and 2 animals (0.5%) had a reaction of 1.3 and 1.9 mm, respectively. The mean increase of skin fold thickness was 0.2 mm. Out of 291 serologically BHV1 positive cattle with an age of more than 6 months 270 had antibodies from natural infection and, partially, from additional vaccination with inactivated BHV1 vaccine. 266 (98.5%) of these animals showed a skin reaction of more than 2.0 mm, in 3 animals (1.1%) a skin reaction up to 1.0 mm was observed and 1 animal (0.4%) had a reaction of 2.0 mm. The mean increase of the skin fold thickness was 6.3 mm. 21 animals had BHV1 antibodies only because of vaccination with inactivated BHV1 vaccine. Only 4 animals had a skin reaction of more than 2.0 mm. Among 107 animals with an age up to 6 months 30 were serologically BHV1 positive and 77 were BHV1 negative. In all animals the skin reaction was less than 1.0 mm, the mean was 0.2 mm.     

Genetic and economic evaluation of a basic breeding programme for Kenya Boran cattle

The objective of this study is to describe the present organisational structure of Boran cattle breeding and develop a model breeding programme using a deterministic approach under the current structure of the Boran Cattle Breeders Society. A breeding unit of 13,000 cows supplying bulls to a commercial population of 39,000 cows was assumed. Selection criteria used were growth and reproduction traits while breeding objective traits targeted were growth, carcass, reproduction, survival, milk yield and feed intake traits. Higher selection intensity was possible for breeding sires resulting in higher genetic gains compared to dams even though selection accuracies were generally low. Annual genetic gains were positive except for dressing percentage, cow survival rate, and age at first calving. However, a reduction of age at first calving by 9.5 days obtained a return of Ksh 49.76. The overall monetary genetic gain obtained was Ksh 84.89 with a profit per cow per year of Ksh 377.98. The basic breeding programme could be expanded to accommodate a larger population of Boran cattle. The implications of the results are also discussed.     

Bovine herpesvirus type 1 (BHV-1) up-regulates telomerase activity in MDBK cells

The proliferative capacity of mammalian cells is regulated by telomerase, an enzyme uniquely specialised for telomeric DNA synthesis. The critical role of telomerase activation in tumor progression and maintenance has been well established in studies of cancer and of oncogenic transformation in cell culture. Experimental data suggest that telomerase activation has an important role in normal somatic cells, and that failure to activate sufficient telomerase also promotes disease. Evidence regarding the role of telomerase in the pathogenesis of several viruses including human immunodeficiency virus has led to an increased interest in the role of telomerase activity in other virus infections. In this research we evaluated the telomerase modulating activity of Bovine herpesvirus 1 (BHV-1) in MDBK cells. MDBK cells were infected at different multiplicity of infection with BHV-1 Cooper strain and telomerase activity at different times post-infection was measured by the TRAP assay. Our data indicate that BHV-1 significantly up-regulates telomerase activity at 3 and 6h post-infection decreasing after the 24h post-infection. Our data, showed that the effect was mediated by an immediate-early or early viral gene, and use of the protein translation inhibitor cycloheximide confirmed that an immediate early gene is primarily responsible.     

Bovine virus diarrhoea (BVD) control programme in an area in the Rome province (Italy)

A BVD control programme based on the identification and removal of persistently infected (PI) animals is being undertaken in an area in the Rome province, where BVD outbreaks had been previously detected. It involves 174 mainly dairy herds, from which blood samples of all bovines older than 1 year are obtained through the national brucellosis and leukosis eradication programme. Samples sufficient to detect the presence of seropositive animals at a prevalence of 5% or more are initially screened for antibodies against BVD virus (BVDV) using an immunoenzymatic assay. Upon identification of seroreagents additional blood samples are tested from the 6-12-month age category not included in the initial samples. Animals are considered immunotolerant if BVDV is demonstrated twice at a minimum 30-day interval. When no seropositive animals are detected during the first serological screening the herd is declared BVD-free if a second testing, preferably carried on the same animals previously tested, confirms the seronegative status of the herd. At present 147 farms have been tested, of which 63 (42.9%) are negative with respect to antibodies against BVDV. Of the 84 remaining herds in which one or more seropositives are detected, 13 are classified as recently infected. In eight of these recently infected herds, 22 PI animals have been identified.     

The detection of bovine herpesvirus type 1 (BHV 1) using an intradermal test. II. Experimental studies

A purified concentrated BHV1 antigen that had been tested in field trials was also tested under isolated conditions in BHV1 positive and negative cattle. The antigen proved to act specifically even after storage for 2 years at +4 degrees C or for 6 months at 37 degrees C. The best results were obtained when the test was read 48-72 hours after the injection, which is in agreement with the field trials. The increase in skin thickness decreased unless boosted by infection or vaccination gradually. The test is unsuitable to control a vaccination programme. There was no correlation between the increase in thickness and humoral antibody titers. Repeated application of the intradermal injection of the antigen did not result in seroconversion in seronegative cattle. The biological limits of the test evaluation are discussed.     

Design of a village breeding programme for a llama population in the High Andes of Bolivia

No national breeding programme for llamas is in place in Bolivia. Initiatives for genetic improvement are rarely found and are usually carried out by NGOs working in rural development or improvement of livestock production or research stations. Farmers in the Province of Ayopaya in the District of Cochabamba have formed a breeders’ organization with the aim of improving fibre production. In this study, a detailed outline of a breeding programme with a focus on organizational and technical details is described. Facing constraints like illiteracy of farmers, bad infrastructure and lack of finances, a simple breeding programme is set up. The breeding goal is a higher fleece weight while keeping the fleece quality at the current high level. Greasy fleece weight and fibre diameter are identified as main selection criteria. Mass selection of males is carried out. Selected males are either exchanged between farmers and used in the herds or are kept during the mating season in a central mating station owned by the breeders’ organization. Model calculations were carried out with the program zplan , which is based on a deterministic approach. zplan evaluates the genetic and economic efficiency of breeding strategies considering one cycle of selection. Scenarios with only intra-herd use, using only the central mating station or combinations of those were compared in terms of expected genetic gain and expected increase of inbreeding. Fastest genetic progress is achieved when the males are kept in a central mating station as the selection intensity is on a high level. Rates of inbreeding vary between 0.08 and 0.32% per generation.     

Risk factors for introduction of BHV1 into BHV1-free Dutch dairy farms: a case-control study

In May 1998, a compulsory eradication programme for BHV1 started in the Netherlands. In December 1999 approximately 24% of Dutch dairy farms were certified BHV1-free (Animal Health Service (AHS)). Ninety-three certified BHV1-free dairy farms participated in a cohort study that investigated the probability of introduction of infectious diseases. The probability of introduction of BHV1 was determined from March 1997 until April 1999. Ninety of these farms remained BHV1-free and could be used as control farms. From January 1997 until March 1998, BHV1 was introduced into 41 BHV1-free dairy farms in the Netherlands (case farms). Management data were collected for both cases and controls and were complete for 37 case farms and 82 control farms. For small data sets and for data in which both low and high frequencies were expected in the contingency tables, the asymptotic methods were unreliable. Our data set clearly resembled such a data set; the risk factors were rare events because the BHV1-free farms were closed farms on which few direct animal contacts occurred. Therefore, an exact stratified modelling approach was most suitable for the data. The study showed that dairy farms should prevent cattle from escaping or mingling with other cattle and that professional visitors should always wear protective farm clothing.     

Prevalence of bovine herpesvirus 1 (BHV-1) infection in Hungarian cattle herds.

Hungarian cattle herds were surveyed for bovine herpesvirus 1 (BHV-1) infection by ELISA of milk and serum samples. In 1993, 75% of the large cattle herds (consisting of more than 50 cattle) and all small herds (small-scale producers' stocks), while in 1997 90% of the small herds were included in the survey. In the case of large herds, 79.3% of the herds and 64.1% of the samples tested were found to be positive. Of the small herds, 13.5% and 15.7% tested positive in 1993 and 1997, respectively. The majority of large herds were Holstein-Friesian dairy stocks. Small herds with an infection rate markedly exceeding the average were found in those counties where the small herds had been in close contact with the large-scale farms, or where new herds were established by using animals of uncontrolled infectious bovine rhinotracheitis (IBR) status originating from large farms. Attention is called to the importance of maintaining the IBR-free status of small herds that constitute one-third of the Hungarian cattle population.