Test of the Einstein Equivalence Principle near the Galactic Center Supermassive Black Hole.

During its orbit around the four million solar mass black hole Sagittarius A* the star S2 experiences significant changes in gravitational potential. We use this change of potential to test one part of the Einstein equivalence principle: the local position invariance (LPI). We study the dependency of different atomic transitions on the gravitational potential to give an upper limit on violations of the LPI. This is done by separately measuring the redshift from hydrogen and helium absorption lines in the stellar spectrum during its closest approach to the black hole. For this measurement we use radial velocity data from 2015 to 2018 and combine it with the gravitational potential at the position of S2, which is calculated from the precisely known orbit of S2 around the black hole. This results in a limit on a violation of the LPI of |ß_{He}-ß_{H}|=(2.4±5.1)×10^{-2}. The variation in potential that we probe with this measurement is six magnitudes larger than possible for measurements on Earth, and a factor of 10 larger than in experiments using white dwarfs. We are therefore testing the LPI in a regime where it has not been tested before.

Bovine immunodeficiency virus and analogies with human immunodeficiency virus.

After describing the results of BIV research during the past years experimental data are presented which indicate that BIV does not cause any clinical symptoms after infection and that no correlation exists with the other widely spread retrovirus in the bovine, the bovine leukosis virus (BLV). Since contact obviously did not lead to a horizontal transmission it is suggested that transmission occurs, as in the cat, vertically from dam to offspring. It was also found that a long period of time after infection can elapse before antibodies against BIV can be detected. It is also quite clear that HIV and BIV do not have much in common except that both are lentiviruses.

Detection and localization of single-base sequence differences in foot-and-mouth disease virus genomes by the RNase mismatch cleavage method.

The RNase mismatch cleavage method was examined for its efficiency of indicating single-base sequence differences in the capsid protein-coding regions of different foot-and-mouth disease virus subtype O1 strains. The method was found suitable for indicating such differences. RNase A as well as RNase T1 contributed to substrate conversion. Examples for the cleavage of eleven different single-base mismatches in RNA double-strands are now known. All virus genomes found to differ from each other exhibited three or more non-neighboured single-base sequence differences. Other genomes found to be indistinguishable by this method were those of a recent field isolate adapted to cell culture, and those of a vaccine production strain; its progeny was transmitted to pig and cow and then analyzed. The results suggest that host change does not necessarily select for antigenic variant virus, and that virus submitted to some kind of selection pressure is changed at more than one position.

Detection of foot-and-mouth disease virus in nasal swabs of asymptomatic cattle by RT-PCR within 24 hours.

A method for extracting RNA from animal-derived materials that provides foot-and-mouth disease viral template suitable for Tth polymerase-dependent synthesis of cDNA and subsequent PCR is described. Viral genomes were detected in less than 24 h. Nasal swabs that can be easily and repeatedly collected, proved suitable for virus detection by PCR, even during the asymptomatic stages of infection.

Studies on the suitability of alpha-hybrid interferon application in cattle.

Twelve cattle with body wts ranging from 100 to 250 kg were treated using various doses and routes for four days with an E. coli derived alpha-hybrid interferon. The lowest parenteral doses (10(4) units per kg body wt) and the orally administered interferon did not lead to any disturbances, whereas the higher dosages led to marked changes in body temperature, pulse and respiration rates. Animals with the highest dose (10(8) units per kg body wt) became extremely distressed. The blood picture showed distinct changes, with very low leukocyte counts during treatment, which took weeks to recover. It is suggested that the dosages that did not lead to clinical symptoms are best suited for prophylactic or therapeutic purposes.

Foot-and-mouth disease. Challenge of cattle after multiple vaccinations.

Four groups of six cattle were vaccinated from two to five times at 6 month intervals with two different trivalent FMD vaccines licensed in the given year. The FMDV type A strains in the vaccines designated A5F and A5B were closely related. Three months after the last vaccination the cattle were challenged by contact with animals inoculated with the original field strain A5B. The inoculated animals developed typical FMD symptoms with vesicles in the mouth and on the feet. Those cattle which had received vaccines that did not contain strain A5B also became severely sick, even after five vaccinations. Animals vaccinated twice with type B containing vaccine were also not completely protected. A safe protection can obviously only be achieved for fairly short periods of time if vaccine and challenge strain are homologous. It is proposed to change the rules of licensing, to speed up the procedure to vaccinate in cases of outbreaks. The need for further research, especially into improving vaccines, is stressed.

BHV1 infections: relevance and spread in Europe.

Infections caused by BHV1 are very common in Europe, but the disease pattern is quite different: the diseases of the genital tract are most common, those of the respiratory tract vary in intensity and prevalence. Digestive disorders connected with BHV1 are in general only observed in calves and mainly in Belgium. Virus strains causing abortion or encephalitis are only present in a few countries. The same is true for BHV1 induced mastitis. Dermatitis and lesions in the interdigital space seem to be a rare event. BHV1 infections are frequently complicated by bacterial secondary infections, but there is evidence that BHV1 infections can occur simultaneously with bovine virus diarrhoea (BVD) and/or parainfluenza-3 (PI 3) virus. The biggest problem associated with BHV1 infection is the ability of the agent to become latent following a primary infection. The genome of the virus probably remains during the life of the animal in the ganglia of the region where the primary infection occurred. No vaccination can overcome this latent stage. By prophylactic vaccination it is possible to prevent an outbreak of clinical disease but it is impossible to prevent infection followed by the establishment of latency. Eradication programmes in Austria, Denmark and Switzerland have removed most of the seropositive cattle from the bovine populations. Currently a sanitary programme is also being conducted in Germany.

Analysis of the phenotype and phagocytic activity of monocytes/macrophages from cattle infected with the bovine leukaemia virus.

The bovine leukaemia virus (BLV) is a retrovirus that infects mainly B lymphocytes of cattle, but proviral DNA can also be isolated from monocytes/macrophages. This study investigated the effect of BLV infection on surface antigens on freshly isolated peripheral blood monocytes and cultured monocyte-derived macrophages, with and without lipopolysaccharide (LPS) stimulation. The effect of BLV infection on phagocytic activity of CD14+ monocytes was also assessed. The percentage of monocytes expressing the surface antigens CD11b, CD32 (FcgammaRII), MHC class II and the surface antigen recognised by mAb DH59B were increased in BLV-positive cattle. In contrast, expression intensity of all markers was low in samples from BLV-positive cattle. CD14+ monocytes from BLV-positive cattle showed less Fcgamma-receptor-mediated phagocytosis compared to monocytes from BLV-negative cattle. After 7 days in culture, there was evidence for shedding/downregulation of surface antigens on monocyte-derived macrophages, in particular on cells from BLV-positive cattle. LPS stimulation decreased the percentage of cells expressing the measured markers in monocyte-derived macrophages taken from BLV-negative cattle, but not in cultures derived from BLV-positive cattle. The results provide further evidence for an altered function of monocytes and macrophages in BLV-infected cattle.

An immunohistochemical study of the distribution of border disease virus in persistently infected sheep.

Three seronegative sheep persistently infected with Border disease virus and six seropositive, non-viraemic sheep were examined for the cellular distribution of the agent. These animals originated from a closed flock which had been kept in an isolation facility for 5 years. They were killed and immediately necropsied. There were no gross abnormalities other than reduced body weight of the persistently infected sheep. Two samples of each major organ were collected. The first sample was fixed by immersion in formalin and processed for histological examination, which showed no lesions unequivocally attributable to the viral infection. The second sample was snap-frozen for immunohistochemical examination. This revealed viral antigen in all organs of the persistently infected, but in none of the seropositive animals. The infected cells included smooth muscle cells of hollow organs and blood vessels, epithelial cells of the alimentary tract and urogenital organs, lymphocytes in lymphoid organs, endocrine cells, neurons and glial cell.

Diagnosis of enzootic bovine leukosis: a comparison of haematological and immunodiffusion tests.

A total of 657 cattle from three different herds were tested for the presence of specific antibodies against EBL antigens by means of the immunodiffusion test and the results compared with the classification according to the lymphocyte counts. All animals with pathologically raised lymphocyte counts showed a positive immunodiffusion test reaction. In the majority of these both antigens, 'p24' and 'gp', gave positive results; in a smaller number antibodies against the gp antigen only were detected, but never against the p24 antigen only. It is concluded that for field testing programmes the use of the gp antigen is sufficient. In future experiments, however, both antigens should be employed, because the lack of one of them might perhaps prove to be correlated with the future course of the disease. The absence of specific antibodies, together with a normal number of lymphocytes, in young animals should, however, not lead to the conclusion that they are free of the disease.

Frequency of surface immunoglobulin bearing blood lymphocytes in cattle affected with bovine leukosis.

Lymphocytes of the peripheral blood from healthy cows and animals suffering from bovine leukosis (persistent lymphocytosis or leukaemia) were examined for the presence of surface immunoglobulin determinants. Using the immunofluorescence technique an average of 18 per cent of lymphocytes from healthy and 67 per cent from leukotic cows could be recognised as positive. An increase of these positive cells, which are supposed to be B cells or of B cell origin, seems to be a characteristic finding in this disease of cattle.

Differences in the in vitro response of lymphocytes from leukotic and normal cattle to concanavalin A.

The response of cultured peripheral blood lymphocytes from cattle affected with bovine leukosis and from animals free of detectable leukosis to the nonspecific mitogen Concanavalin A ws determined by the measurement of 3H-thymidine incorporation. The stimulatory effect of the mitogen was significantly decreased in leukotic lymphocytes in comparison with normal lymphocytes. This depression was dependent on the severity of the disease. Leukotic lymphocytes showed high basal levels of spontaneous thymidine uptake possibly due to their low maturity. It is suggested that leukotic lymphocytes are not stimulated by Concanavalin A because of their B cell characteristics. A residual cell population of normal reacting lymphocytes--enriched by nylon wool column fractionation--is probably responsible for the remaining but weak in vitro reaction of leukotic lymphocytes.

Some observations on the epidemiology of bovine leucosis virus infection in a large dairy herd.

Bovine leucosis infection rates were calculated for two years in a naturally infected dairy herd in which serologically positive animals were not preferentially culled. Transmission of infection was found to occur mainly during the winter housing period. No variation in susceptibility to infection with age was found and young animals did not show a prolonged time from infection to sero-conversion.

The bioavailability of supplemental biotin in cattle.

A trial using 12 yearling heifers was carried out to test whether biotin metabolism and bioavailability are influenced by continuous dietary supplementation with biotin. Six of these heifers received no biotin supplementation (controls), while six received a daily dietary supplement of 20 mg biotin over the whole experimental period of four months. During each of three test periods (on days 14 and 21, 56 and 63, and 118 and 124), single test dosages of 40 mg (oral) and 5 mg (intravenous) biotin were given to each animal in a crossover test design. Blood samples were collected up to 72 h after each of these single doses, and at approximately two-weekly intervals for the assessment of baseline values. Serum biotin levels were determined by an ELSA test. Areas under the curves (AUC) were calculated as the target parameter for the assessment of the bioavailability of orally administered biotin. Serum biotin baseline levels were 300-800 ng/l in the controls and 3000-8000 ng/l in the supplemented animals. In both groups, AUC values in the first test period (days 14 and 21) were significantly higher than in subsequent periods. However, the biotin supplementation showed no significant effect. There was no significant difference in elimination half-lives between groups with and without biotin supplementation. The range was 5-18 h. Furthermore, there was no significant difference in the bioavailability of biotin between the test periods or between the biotin-supplemented and unsupplemented animals. Overall bioavailability was 48%.

Biotin kinetics in serum of cattle after intravenous and oral dosing.

Single oral (p.o.) or intravenous (i.v.) doses of biotin were given to four cattle (400-450 kg body weight) in two consecutive tests two weeks apart. Dosages were p.o. 20, 40, 80 or 160 and i.v. 5, 10, 20, 40 mg biotin per 300 kg body weight. A three-compartment model was used to describe the course of serum concentrations with time. After i.v. administration, terminal half-lives of about 8 h were found. Areas under the curves were linearly related to both the p.o. and the i.v. doses. The systemically available fraction of the p.o. dose was 50 to 60%. On the basis of kinetic parameters, the biotin uptake via the feed was estimated to be 2.5 mg/day, which was about half of that estimated to be in the hay consumed. The data suggest that there was no relevant ruminal synthesis of biotin.

Vaccination to protect calves against infectious bovine rhinotracheitis.

A commercially available inactivated vaccine against infectious bovine rhinotracheitis (BHV1) was tested to assess its ability to immunise young seronegative calves and protect them against challenge with a virulent strain of BHV1. Calves showed seroconversion after one or two doses of vaccine. A two-dose and three-dose vaccination regimen each afforded calves significant protection against challenge as judged by the development of clinical symptoms. Vaccinated calves were on average 7 to 10 kg heavier than control calves 24 days after challenge, a statistically significant difference. Vaccination had no significant effect on the virus excretion pattern after challenge.

Risk of BSE from the import of cattle from the United Kingdom into countries of the European Union.

This study assesses quantitatively the risk that other countries, in particular those within the European Union, have incurred by importing cattle from the United Kingdom during the period before or shortly after the ban on the import of live breeding stock was introduced in 1989. It does this by assessing the probability that animals imported from the UK in a certain year would have become a detected BSE case, had they not been exported. Using the annual incidence rates available for separate birth cohorts and a given culling rate, a cumulative incidence for each birth cohort was calculated. These figures were then combined with the numbers of live breeding cattle imported from the UK into the other countries of the EU, to give an import-related risk index for each country, assuming that their culling rates were similar to that in Great Britain. The countries could thus be categorised in terms of the number of cases of BSE they might have expected.

[Viral agents as the cause of reproductive disorders in cattle and available vaccines]. / Virale Erreger als Ursache von Fortpflanzungsstörungen beim Rind und verfügbare Impfstoffe.

After a review on the viral agents playing a role in diseases of cattle those related to the occurrence in the genital tract are described. They may be causing abortion or local reactions leading to a reduced fertility and/or be of importance for the embryo transfer. Bovine herpesvirus type 1 (BHV1) and the bovine virus diarrhoea virus (BVDV) are the agents most widely distributed in Europe. Both are of economic importance, described in detail and vaccines available discussed.

Advances in BHV1 (IBR) research.

Bovine Herpesvirus Type 1 (BHV1) is the aetiological agent of a number of diseases and not only of IBR, namely infectious pustular vulvovaginitis (IPV), infectious balanoposthitis (IBP), conjunctivitis, encephalomyelitis, mastitis, abortion, enteritis, and lesions in the interdigital space. The serological identical strains differ, however, in some aspects. Typical genital strains usually cause a mild illness, sometimes not even detected clinically, but serologically. They hamper eradication programmes and do not cause IBR when inoculated intranasally. The other--modern--strains are, however, always able to induce a severe disease in the genital tracts. But infection of field or vaccine virus leads to the development of humoral and cell-mediated immunity. The latter is, however, not transmitted to neonates via colostrum. BHV1 antibodies can be found in bovines in all continents, and in many wild species. Prevalences vary greatly depending on herd size and management. Because seronegative cattle play a role in international trade a number of European countries have eradicated BHV1, with very high costs involved. Marker and conventional vaccines can prevent disease but not infection followed by the state of latency. The genomes of several strains, including the marker strains can remain latent in the same animal and be reactivated after stress or injection of corticosteroids. For the detection of humoral antibodies the ELISA is widely used. It is useful for testing bulk milk samples for antibodies derived from field virus and conventional vaccines but not from gE-deleted marker vaccines. Importing countries should consider only vaccinated animals for import. They should require that the animals are seronegative prior to vaccination.

[Foot-and-mouth disease. Infection trial in cattle after a single vaccination]. / Maul- und Klauenseuche. Infektionsversuch an Rindern nach einmaliger Impfung.

Following an outbreak of FMD caused by an A5 strain in the spring of 1984, ten cattle were vaccinated with samples of the five commercial vaccines used for the vaccination campaign in that year, i.e. two animals per vaccine. Six weeks later the cattle were challenged by contact with animals inoculated with the virus strain isolated from the field outbreak. Seven of the ten cattle became severely ill, exhibiting the typical symptoms of FMD, one animal did not show any clinical symptoms, the remaining two weak ones that might have escaped recognition by the cattlemen. Virus could be recovered from the vaccinated animals from days 2 to 10 following contact with the non-vaccinated infected cattle. It was concluded that a single vaccination does not protect cattle against the isolate.