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A modified method for autoradiographic determination of antibodies to the virus of transmissible gastroenteritis of swine was developed. It is based on the actual reaction between antigen bound in cells of the infected cell cultures and antibodies in tested sera, which is visualized, by help of rabbit antibodies marked 125J (125J RaSw IgG antibody) to porcine IgG, on a sensitive radiograph and evaluated on the basis of darkening at the point of positive immunological reaction. Specificity of the test and mutual comparability and reproducibility of the results were confirmed by examining the known positive and negative sera and by comparison with the results of the virus-neutralisation test. Out of the 36 examined porcine blood sera the antibodies were proved autoradiographically only in the samples positive also by virus-neutralisation. In experimentally infected pigs, the same dynamics of antibody production was recorded by the two tests. They were, however, demonstrated autoradiographically already by the two tests. They were, however, demonstrated autoradiographically already the eighth day after infection, while by virus neutralisation test as late as 14th day. Their level increased gradually until 35th day after infection.

The rise and mass incidence of intestinal and respiratory infections in calves kept in large herds depend on a joint influence of infective agents and numerous endogenous and exogenous factors, mostly with non-specific action. Therefore, purely medical approach fails to provide efficient prevention. Detailed epizootological analysis is needed for taking actual measures; the analysis should concern potential respiratory and intestinal pathogens and should also cover the persistent infections such as IBR, BVD-MD and others. The immunological profile of herd, very unfavorable with the high culling rate, will improve as a result of the introduction of a purpose-oriented vaccination programme, higher level of colostral nutrition, and separate rearing of calves, isolated from the dams, in calf houses managed by the all-in-all-out system.

For evaluating the protective effect of polyvalent vaccine against diarrhoea in new born calves caused by rotaviruses, coronaviruses and enterotoxigenic E. coli the method was selected of mathematical and statistical analysis of the set of data characterizing the most important clinical symptoms of the disease during the first three weeks after birth. In two large-scale breeds with a mass occurrence of diarrhoea of the known etiology the state of health of calves before the vaccine application was compared with the state of health of calves born in the period after the vaccination. The following traits were selected for evaluating the severity of the disease: the age of calf at the beginning of the disease, character and intensity of diarrhoea, length of disease and results of treatment. In all the selected indicators, statistically significant differences were determined pointing to the fact that the vaccine used for increasing the immunity against enteropathogenic viruses and bacteria, applied in high pregnant cows and heifers before parturition, protected the sucking calves and calves fed the milk of their mothers against the disease in the first week after birth. The diarrhoea, occurring in the calves of the vaccinated cows and heifers mostly during the second week of age was easy to treat therapeutically and ended in most cases after a short period of a tea diet. After the vaccination, the death loss in both localities decreased more than eight times.

The most frequent microbial causative agents of massive diarrheas in new-born calves kept on large cattle farms in the CSSR are rotaviruses, coronaviruses and enterotoxigenic strains of E. coli, manifesting themselves as complicated virus-bacterial infections. An inactivated polyvalent adjuvant vaccine has been developed for the prevention and specific prophylaxis of these enteral infections; the vaccine contains bovine rotavirus, bovine coronavirus and three enterotoxigenic serotypes of E. coli with protective antigen K 99. The rotavirus and coronavirus are propagated on the stable cellular line MDBK and inactivated with 0.2% formalin, the Escherichia strains are submersed in the MINCA culture medium during their cultivation and inactivated with 0.5% formalin. The vaccine was prepared as a blend of the same amounts of rotavirus and coronavirus and of such an amount of bacterin so that 1 ml of the vaccine will contain 10(9) bacteria. One part of oil adjuvant was added to five parts of the virus-bacterial blend and the blend was homogenized in the Ultraturax apparatus. The vaccine is to be used for immunization of pregnant cows and heifers; in these animals it induces the production of specific antibodies to all antigens contained in the vaccine. Its immunogenic effects were checked in 32 calves and 38 cows in the herds with the occurrence of diarrheas caused by both enteropathogenic viruses and enterotoxigenic escherichia. It was demonstrated that the inactivation did not influence in either of the viruses the process of inducing the production of specific antibodies, and the antibody response of the calves and heifers after application of 2 ml of complete inactivated vaccine was equally strong as after application of live vaccine containing only rotavirus and coronavirus. The level of the rotavirus antibodies increased on the average 30 times and 200 times, coronavirus antibodies twice and four times. The antibody response to coronavirus was negatively influenced by the relatively high levels of antibodies before vaccination. The antibody response to antigen K 99 was expressive in all cases.

Bovine coronavirus isolated from calf faeces diseased with gastroenteritis and passaged to colostrum-free calves agglutinated mouse and rat erythrocytes. The agglutination reaction depended on temperature and took place only at a temperature of 4 degrees C. At a temperature of 37 degrees C the agglutinate broke down within 15 minutes. The coronavirus could be detected by the haemagglutination test in the contents of the small and large intestines and in the faeces of experimentally and naturally infected calves. The agglutination capacity of mouse erythrocytes was not affected by careful fixation of these erythrocytes with formalin and subsequent lyophilization and remained unchanged for as long as 52 weeks of storage at a temperature of 4 degrees C. It was demonstrated by a comparative examination of 182 samples of the faeces of calves suffering from diarrhoea that haemagglutination test was as sensitive as electron microscopy.

It is not easy to exactly diagnose the etiology of the mass infections of new-born calves on large farms where considerable losses are suffered. On the basis of the complex epizootological, clinical and laboratory examination in four large calf-rearing facilities, rotaviruses, coronaviruses, the infectious bovine rhinotracheitis (IBR) virus and the bovine viral diarrhoea (BVD) virus, and in some cases also the enteropathogenic E. coli, were found to be etiologically involved in the mass rise of diarrhoea, complicated by respiratory symptoms already during the first days after birth. The clinical picture of the disease, therapeutically difficult and reminding of "pneumoenteritis", has often been observed in stocks where, in addition to rotaviruses and coronaviruses in the faeces, the IBR or BVD viruses (sometimes both at the same time) were detected and identified in the respiratory and enteral tract. The serological examination of a higher number of animals in the stocks of calves under study confirmed the considerable rate of spreading of all the four viruses in the cattle population and, at the same time, demonstrated the very unfavourable immunological profile of the herds. The high percentage of animals low in antibody titres and the serologically negative animals constitute the infection-sensitive part of population in the affected herds. With the high culling rate and with the open herd turnover it is impossible to reach the required immunity through natural disinfection. Loss-free rearing of healthy calves will be achieved on the basis of a well-oriented vaccination programme with a good combination of inoculants.

There is a description of the enzymoimmunologic method (ELISA), which was used for demonstration of rotaviruses and coronaviruses in the samples of excrements of the calves suffering from diarrheas. It is shown by a comparison with the results obtained by electron microscopy that the ELISA method provides by up to 50% higher capture rate and the reaction is highly specific. The method can also be applied to a detection of human rotavirus in the children's stool.