PCR for detection of Mycobacterium tuberculosis in experimentally infected dogs.

The susceptibility of dogs to infection with Mycobacterium tuberculosis was studied through different ways of experimental infection. The examination shows, that in most cases the disease runs subclinically with pathological changes localized mainly in the lungs, lymph nodes, small intestines, liver, kidneys and spleen. Histological findings demonstrate granulomatous inflammation with caseosation and predominance of epitheloide macrophages and single lymphocytes. Tissue samples from internal organs of experimentally infected dogs as well as non-infected but contact animals were investigated by direct PCR. Specific PCR-products were obtained in 44 of 96 studied samples. Eighty-three (86.5%) of PCR results coincided with bacteriological finds, 82 (85.4%) with the pathological and 71 (74.0%) simultaneously with bacteriological and pathological results. The observed specific DNA products in tissue samples of infected and non-infected dogs demonstrate significant sensitivity of PCR method. It could be assumed that the transmission of M. tuberculosis infection is possible by close contact between ill and healthy dogs and that the naturally infected dogs or dogs suffering from tuberculosis may serve as a permanent source of infection to humans and other animals.

[Drug resistance and virulence of Staphylococcus aureus strains isolated from cows with endometritis]. / Lekarstvena ustoichivost i virulentnost pri shtamove Staphylococcus aureus, izolirani ot kravi s endometrit.

A study was carried out on the drug resistance as associated with the virulence of Staphylococcus aureus strains isolated from cows with endometritis. Hirota's method was employed to ascertain that resistance to penicillin, erythromycin, neomycin, and kanamycin was governed by plasmids. It was found at the same time that, fairly often, with the elimination of some of the resistant markers the virulence of the strain variants also dropped. However, these two properties did not correlate with each other.

[Effect of levamisole on the cytochemical function of leukocytes and on blood lysozyme in sheep]. / Vliianie na levamizola vurkhu funktsionalno-tsitokhimichnoto sustoianie na levkotsitite i na lizotsima v kruvta na ovtsete.

It was established in an experiment with 13 sheep of the local improved breed that the single s/c injection of levamisole at the rate of 7.5 mg per kg of body mass led to a rise of the phagocytic activity of neutrophil leukocytes. These cells also had higher cytochemical activity of the alkaline phosphatase, glucose-6-phosphatase, and lipids (sudanophilia). No changes in the total count of leukocytes were found. The cytochemical activity of lactate-dehydrogenase, succinate-dehydrogenase, and alpha-glycerophosphate-dehydrogenase was also found to rise. On the base of the marker capacity of the acid naphthylacetate esterase with regard to the T-lymphocyte system there set in on the 3rd day following levamisole injection a 4 to 5 rise in the T-lymphocyte count, particularly in the mature T mu-lymphocytes, to the detriment of the B-cells which dropped in number. On the 7th day all these changes receded, however, the percent of lymphocytes with azure granules in the cytoplasm and the average number of these granules per lymphocyte cell rose. In the entire 7-day period of investigation a lowering trend was shown by the amount of serum lysozyme which dropped several times as against the initial level.

[Effect of levamisole on white mice and on the immunological reactivity of guinea pigs]. / Vliianie na levamizola vurkhu beli mishki i imunologichnata reaktivnost na morski svincheta.

Investigations were carried out on the protective action of therapeutic doses of levamisole (Pharmachim) on mice with a LD100 Salmonella gallinarum infection. Studied was also the effect of levamisole on the immunologic response of guinea pigs infected with Salmonella cholerae suis. It was found that therapeutic amounts of 3 mg/kg levamisole applied to mice following respective patterns protected them against a lethal dose of S. gallinarum. The animals showed enhanced resistance to the infection--20 to 50 per cent of them survived as against 100 per cent mortality with the controls. The guinea pigs showed upon treatment with levamisole certain changes with some of the factors of unspecific response--enhanced phagocytic and lysozyme activity as judged by the development of infection with a S. cholerae suis culture. At the same time the agglutinin titer that substantiated the immunologic response was negligibly changed as compared to the controls. On the base of the results obtained an attempt was made to elucidate the mechanism of the protective action of levamisole.

[Protein studies of viral hemorrhagic septicemia in rainbow trout]. / Izsledvaniia vurkhu beltuchinite pri virusnata khemoragichna septitsemiia pri dugovi pusturvi.

Comparative investigations were carried out on the changes in the blood serum of normal and diseased rainbow trout with virus hemorrhagic septicaemia. Determined was the amount of the total protein and the protein fractions. The electrophoretic investigation of blood serum of rainbow trout revealed one dominating albumin fraction, 3 alfa-globulin fractions, 2 beta-globulin ones, and one gamma-globulin one. The following up of the protein fraction dynamic during the various seasons did not reveal any dependence on the temperature curve. Rainbow trout manifesting clinically virus hemorrhagic septicaemia showed lowering of the total protein amount and changes in the electrophoretic profile and the quantitative ratio of the protein fractions. There were a drop of the albumin percent and a rise of the beta- and gamma-globulin fractions. This led to a change in the coefficients alfa:beta (0.89) and albumin: globulin (0.14).

[Protein fraction changes in swine raised under different conditions]. / Promeni v beltuchnite fraktsii u svine, otglezhdani pri razlichni usloviia.

The total protein and the protein fractions were studied in pigs raised both under industrial conditions and on the personal farms. It was found that the process of immunogenesis in pigs varied largely parallel to the various conditions of raising. The numerous immunoprophylactic treatments under industrial conditions were found to have an adverse effect on the forming of the immune response, even without apparent antigen competition. The investigations aimed at elucidating the expediency of the adopted and very often unsubstantiated, repeatedly effected treatments for the rational use of mixed vaccines.

[DNA nucleotide composition of Gram-positive cocci isolated from food products of animal origin]. / Nukleotiden sustav na DNK ot grampolozhitelni koki, izolirani ot khranitelni zhivotinski produkti.

Studied was the nucleotide composition of 34 strains of gram-positive cocci isolated from slaughtered birds, sheep milk, and milk from mastitis-affected cows and ewes. The differentiation of the strains was carried out in conformity with the instructions of Bergey's Manual of Determinative Bacteriology (1974) on the basis of the following indices: morphology of colonies, morphology and staining of the cells after Gram, catalase, oxydase, respiration type, fermentation of manite, production of plas mocoagulase, thermonuclease, and phosphatase and response to novobiocin. The amount of guanine, adenine, cytosine, and thymine in the investigated strains was determined after the method of Spirin and Belozerskii. On this basis the guanine-cytosine percent was calculated. Results showed that the use of the guanine-cytosine percent made it possible to more accurately differentiate the genus with gram-positive cocci. In 88.24 per cent of the investigated strains there was coincidence between the guanine-cytosine percent and their biochemical characteristics. Two of the strains (guanine-cytosine percent 49.95 and 47.48) were shown to belong to the group of planococci. No typical micrococci (with a guanine-cytosine percent of more than 60.00) were established.

[Determination of the nucleotide composition of Pasteurella multocida strains]. / Opredeliane nukleotidniia sustav na shtamove Pasteurella multocida.

Studied was spectrophotometrically the nucleotide composition of DNA of Pasteurella multocida strains along with studies on the nucleotide base ratio, the molar relations between nucleotide bases, the guanine/cytosine ratio in percent, and the specificity index adenine-tinin/guanine cytosine. The strains used were isolated from birds with various forms of fowl cholera (acute, atypic, and swelling of the wattles), and from calves with bronchopneumonia. Used were also mutants that were streptomycin dependent, obtained through induction with nitrosoguanidine. Close values were established of the nucleotide bases both with the individual groups of Pasteurella strains and with the mutants and their initial strains. The molar relations showed values around a unit, the guanine/cytosine ratio in percent varied from 40.14 to 43,54 and the specificity index varied from 1.25 to 1.49. Data showed that all studied strains and mutants belonged to one species - Pasteurella multocida, on the one hand, and their grouping could not be made on the basis of their nucleotide composition, on the other - contrary to some other indexes, such as biochemical activity, virulence, and behaviour to specific phages.

[Trails with levamisole to stimulate immunity against Newcastle disease in poultry]. / Opiti za stimulirane na imuniteta sreshtu psevdochuma po ptitsite s levamizol.

Attempts to stimulate the immunity against Newcastle disease in birds aged 2 months, of the Leghorn breed, were carried out. It was found that levamisol at the rate of 3 mg/kg body weight given once, parenterally, 24 hours prior to or after the immunization of the birds with a live La Sota vaccine or given three times, parenterally or orally, did not stimulate the humoral immune response. No differences were found in the level of the total protein, protein fractions, and the titer of the hemagglutinating antibodies between the test and the control birds. There was, however, prolongation of the time of effective immunity in the levamisol-treated and vaccinated birds as against the birds that were vaccinated against Newcastle disease only. Discussed in the possible mechanism of stimulating the immunity with levamisol.

[Elimination of multidrug resistance in E. coli in calves in vivo with rimactan]. / Eliminirane mngolekarstvenata rezistentnost na E., coli pri teleta in vivo s rimaktin.

An experiment was carried out for eliminating the multimedicinal resistance markers of E. coli, populating the intestinal tract of calves, in vivo with rimactan introduced per os, and rationed 10 mg/kg of live weight, once during a period of 8 days. The highest percentage and the longest elimination were observed for the neomycin, the novobiocin and the chlornitromycin resistance markers. The elimination was weaker for the erythromycin, the streptomycin and the kanamycin markers and the weakest was for the penicillin and tetracycline markers. There appeared a difference in the elimination of the resistance markers with the different calves, especially for the markers with a low degree of elimination, depending on the individual peculiarities of the calves. Riphamycin proved to be an eliminating means for the resistance markers of E coli in vivo of calves suffering from enteritis. Alongside with the elimination of the resistance markers, due to the treatment of calves with rimactan, an almost complete recovery was achieved. Rimactan is a reliable means for fighting enteric illnesses with calves, caused by enteropathogenic E. coli.

[Attempt at eliminating the multiple drug resistance of E. coli in pigs with enteritis using Rimactin]. / Opit za elimnirane s rimaktin mnogolekarstvenata ustoichivost na E. coli pri praseta s enterit.

The eliminating effect of rimactan was studied in vivo on resistance markers of E. coli, isolated from 18 new-born pigs with a clinic of enteritis. Rimactan is given per os in 15 mg/kg, liver weight, once a day in the course of 6 days. The sensitivity of the strains eliminated was checked in vitro in respect of 16 medicinal preparations (Pe, Sm, Km, Neo, Chl, Novo, Te, Er, Ty, Sp, Le, Am, Ox, Oxte, Ge-penicillin, streptomycin, kanamycin, neomycin, chlornitromycin, novobiocin, tetracycline, erythromycin, tylan, spectam, lentamycin, ampicillin, oxacillin, oxytetracycline, gentamicin and borgal). To 11 of them E. coli were resistant. After a treatment with rimactan an elimination of resistance markers was observed right on the first day, namely, with regard to Sm, Chl, Novo, Te, Er, Sp, Oxte. On the second day was eliminated the Pe-marker, on the third--the Ty-marker, and it was not until on the fifth day that Am and Ox-markers were eliminated. The elimination frequency was the highest between the third and the fifth days. The experiments studied also the sensitivity of the investigated coli strains with regard to different rimactan concentrations (2-256 mg/cm3) in vitro. It was most pronounced for a concentration of 16-32 mg/cm3. It was proved that rimactan can be used as a preparation for eliminating resistance markers (R-factors) of E. coli in pigs suffering from enteritis.

[Amino acid metabolism in Pasteurella multocida studied using thin-layer chromatography]. / Prouchvane na aminokiselinnata obmiana pri Pasteurella multocida chrez izpolvane na tunkosloinata khromatografiia.

Amino asid metabolism was studied by thin layer chromatography in 25 Pasteurella multocida strains isolated from various manifestations of the disease chicken septicemia. Six amino acids were used: DL-ornithin, L-cystein, L-asparagin, DL-serin, L-arginin and Acidum glutaminicum. The Pasteurella strains studied include in their metabolism the amino acid asparagin, but serin and cystein are excluded. The strains isolated from birds suffering from acute and atypical septicemia do not metabolize ornithin, while these isolated from birds having wattles oedema or from calves suffering from arthritis metabolize it. The reaction of Pasteurellae to arginin and glutamic acid varies and no regularity in this respect is observed. Strains isolated from calves suffering from bronchopneumonia do not include in their metabolism any one of the named amino acids. As a result of the test applied a correlation between the Pasteurella multocida biotypes assessed by the authors and the amino acid metabolism was established. This is one proof more that the isolated pasteurellae can be groupped in two big biotypes by the tests for virulence in birds and including in their metabolism the amino acid ornithin.