A dynamic study of drug resistance in populations of faecal Escherichia coli from calves fed Nitrovin and furazolidone continuously.
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Thirty-two different types of anaerobic bacteria isolated from chickens have been tested for anti-salmonella activity in vitro. Under the conditions of the test only Bacteroides hypermegas and a Bifidobacterium sp. were shown to inhibit the salmonellas and this was attributed to the production of volatile fatty acids (VFA's) coupled with a low pH. When these organisms were tested in newly hatched chicks no inhibition of S. typhimurium occurred. Possible explanations for this observation are considered. The pH value and concentration of VFA's in the caecal material were determined in chicks from 0-84 days. In vitro tests with S. typhimurium indicated that, whilst the organism would be able to multiply at the pH and concentration of VFA's found during the first few days after hatching, the rapid increase in VFA concentration during the first 21 days would make this increasingly difficult. The significance of the developing caecal flora in relation to VFA production and pH is discussed. Because certain feed additives are known to influence the carriage of salmonellas, the sensitivity of various caecal anaerobes to these compounds was determined in vitro, generally at 1, 10 and 100 microgram/ml. The additives tested included flavomycin, furazolidone, nitrovin, tetracyline, tylosin, sulphaquinoxaline, virginiamycin and zinc bacitracin. All the organisms tested were inhibited by 100 microgram/ml furazolidone; none were inhibited by 500 microgram/ml sulphaquinoxaline. Changes occurring in the VFA concentration, pH value and microflora of the caeca of chicks fed for 49 days or longer on a normal starter diet or the same diet containing 10 or 100 mg/kg nitrovin have been compared. When the chicks were fed on the diet containing 100 mg/kg nitrovin, the Gram-negative non-sporing anaerobes were eliminated as a significant part of the caecal flora. However, the VFA concentration combined with a low pH in chicks from 2 weeks onwards was still sufficient to inhibit salmonella multiplication. Other possibly interrelated factors which might lead to an increased salmonella carrier rate in the nitrovin-treated chickens are discussed.
The effect of growth-stimulating agents (15 ppm of virginiamycin, 10 ppm of avoparcin and 12 ppm of nitrovin) and that of seven days' treatment with 300 ppm of furazolidone on the performance of broilers was studied in an experimental study of 9,600 animals. Furazolidone was fed in either the fourth or the fifth week of life and combined with virginiamycin or nitrovin. Non-significant improvements in growth were observed when virginiamycin (1.5 per cent), avoparcin (0.7 per cent) and nitrovin (0.7 per cent) were administered. Treatment with furazolidone for seven days resulted in substantial retardation of growth. When broilers were treated during the fourth week of life, the retardation of growth was largely compensated for during the other two weeks. Treatment in the fifth week of life resulted in lower weights prior to slaughter. Therefore, care should be taken in recommending treatment with furazolidone at the end of the fattening period of broilers.
The stimulative effect of Carbadox, Nitrovin and Glypondin substances was tested on juvenile rats. A stimulative effect relative to physical weight was proved only with Carbadox at a dose of 2 mg kg-1 and Nitrovin (only in males) in doses of 10 and 50 mg kg-1 p. o. An overdosing of Carbadox revealed rather a negative effect, because already a dose ten times higher had a negative effect on physical growth and a dose fifty times higher caused obvious toxical symptoms. Glypondin was ineffective in each of the tested doses. The consumption of food per unit of weight increase was reduced after supplying carbadox dosed at 2 mg kg-1 by 25.6% in males and by 10.8% in females. After Nitrovin the intake of food was reduced most after a dose of 10 mg kg-1 in both sexes (8.6 and 8.4% respectively). The most frequent effect on the change of physical weight was the relative reduction in weight of the thyroid gland. For the screening of stimulators laboratory rats are not very suitable.
Groups of 45 chickens were fed continuously on diets containing 10 or 100 mg./kg. of different 'growth-promoting' antibiotics and infected orally with a nalidixic acid-resistant mutant of Salmonella typhimurium. The amount of S. typhimurium organisms excreted in their faeces was estimated by culturing them at weekly intervals and in a standard manner on plates of brilliant green agar containing sodium nalidixate, both direct and after enrichment in selenite broth. All of four groups fed diets containing 100 mg./kg. of nitrovin in three different experiments excreted much larger amounts of S. typhimurium than did groups fed antibiotic-free diets. In some, but not all, experiments, larger amounts were also excreted by groups fed diets containing 10 mg./kg. of nitrovin or 10 or 100 mg./kg. of flavomycin or tylosin. Feeding diets containing 10 or 100 mg./kg. of virginiamycin or bacitracin either did not influence or slightly increased the amount of S. typhimiurium excreted. Two groups fed continuously on diets containing 100 or 500 mg./kg. of sulphaquinoxaline for 44 days excreted smaller amounts of the S. typhimurium organisms that did groups fed antibiotic-free diets; no sulphonamide-resistant organisms of the S. typhimurium strain were isolated from the faeces.
Groups of 33 chickens were fed continuously on diets containing feed additives that are employed commercially for a variety of purposes, and were infected orally when 4 days old with a nalidixic acid-resistant mutant of Salmonella typhimurium. The amount of S. typhimurium organisms excreted in their faeces was estimated by culturing them at intervals and in a standard manner on brilliant green agar containing sodium nalidixate; when the chickens were killed their caecal contents were examined by the same technique.Avoparcin and lincomycin, like nitrovin and tylosin (Smith & Tucker, 1975b), favoured colonization of the alimentary tract by the S. typhimurium organisms as shown by the fact that the chickens to which they were fed excreted these organisms in their faeces in higher concentration and for longer periods of time than did chickens fed on non-medicated diets. Amprolium, monensin, dimetridazole, arsenilic acid and nitro-hydroxyphenylarsonate had no obvious effect on the salmonella excretion pattern.When only five chickens in each group were experimentally infected so that the effect of the feed additives on infections acquired by contact could be monitored, avoparcin, lincomycin, nitrovin and tylosin again favoured colonization of the alimentary tract with the S. typhimurium organisms and so did dimetridazole. Arsenilic acid, in contrast, hindered the development of infection. Amprolium, monensin and nitro-hydroxyphenylarsonate were without obvious effect.Many of the chickens that were fed on diets that favoured S. typhimurium colonization, but not those fed on non-medicated diets, were still excreting S. typhimurium organisms in their faeces when they were killed at 56 days of age, the age at which broiler chickens kept under commercial conditions are usually slaughtered.
Day-old cockerels were used to examine the growth responses obtained when 50 ppm procaine penicillin, 25 ppm zinc bacitracin, or 20 ppm nitrovin were administered in a wheat-based diet. The birds were fed ad libitum from one day of age a plain or supplemented diet in a series of 10, 21, and 56-day tests conducted between 1971 and 1974. Growth responses, measured by improvement in the live weight gain and feed conversion, were obtained with each treatment, consistently over each annual period. The penicillin and zinc bacitracin gave maximal differences in live weight gain compared with the controls before 14 days of age and, possibly, before 7 days of age, whereas the maximal response to treatment with nitrovin occurred after 14 days of age.
In two short-term experiments (28 and 31 days, respectively), the ergotropic effect was tested of the growth promoters nitrovin (25 mg/kg mixture, and a chinoxalin-1,4-N-dioxide derivate (25 and 50 mg/kg mixture). The experiments were carried out on a total of 1,952 unsorted one-day-old White Leghorn chicks under tropical conditions in Eastern Ethiopia (College of Agriculture at Alemaya). On addition of 50 mg chinoxalin-1,4-N-dioxide, the final weight of the chicks was 105.6 and 104.6%, respectively, in the first and second experiments as compared with the unsupplemented groups. The amount of feed required for each weight gain was reduced to 94.0 and 91.4%, respectively. Nitrovin and smaller amounts of chinoxalin-1,4-N-dioxide increased the body weight to 104.2 and 104.0%, respectively, as compared with the control. The feed requirement (94.6 and 99.9%, respectively) was not reduced significantly (alpha greater than 0.05). Further experiments will be necessary, using other feed additives, more chicks, and longer experimental periods.
Two experiments were carried out with sows for fattening having a body weight of 20 to 100 kg. The following nutritive antibiotics were tested -- bacipharmin -- 10% (PRB), zinkbacitracin -- 10% (UFRJ), vitamycin 3 (USSR), nitrovin (Peizun, USA) and mekadox -- 5% (USA). It was established that the stimulating effect of these antibiotics was manifested during the first fattening period -- up to 60 kg body weight, when the average daily increment of the experimental groups treated with bacipharmin, zink bacitracin and nictrovin in the I experiment was 7--15% higher and in the II experiment in which zink bacitracin, vitamycin 3, nitrovin and mekadox were added -- 8--14% higher as compared to the control groups. During the second period of fattening (60--100kg) no effect of the same nutritive antibiotics was observed, with the exception of the vitamycin treated group. In both experiments an economy of combined fodder per 1 kg body weight was realized in the experimental groups. Application of nutritive antibiotics and biostimulators during the second period is economically unexpedient. The antibiotics tested have no negative effect on carcass slaughter characteristics and on the chemical composition of meat. No residual substances of the nutritive antibiotics tested were detected on the second day following slaughter in the meat group treated with mekadox, since no sows of this group were slaughtered.
The antimutagenic properties of whole fresh-water blue-green algae Aphanisomenon flos-aquae, marketed under the commercial name "Alpha Sun" were tested using the Ames test. Simultaneous addition of both algae and Nitrovin (a mutagen) to the test medium did not reduce the mutagenic activity. On the other hand, addition of freeze-dried blue-green algae to the test medium 2-24 h before the application of mutagen reduced its mutagenic activity.
The diethyl ether extracts isolated from unfermented milk and milk fermented by Enterococcus faecium exhibited dose-dependent inhibition of mutagenesis induced by N-methyl-N'-nitro-N-nitrosoguanidine (MNNG), nitrovin (NIT), 5-nitro-2-furylacrylic acid (NFA) and UV-irradiation on the Ames bacterial test (Salmonella typhimurium strains TA97 and TA100) and the unicellular flagellate Euglena gracilis. Overall, the fermented milk extract was the most active against UV-irradiation, less active against NIT and MNNG, and the least active against NFA on bacteria. The highest antibleaching effects were observed against MNNG. The differences between antimutagenic effects from fermented and unfermented milk extracts were determined to be statistically significant at the 0.95 CI level.
2,3-dimethyl-quinoxaline-1,4-dioxide and 2-methyl-quinoxaline-1,4-dioxide proved to be ergotropic in the feeding of a hydrocolloid ration as well as of a broiler fattening feed. They are at least equal to Nitrovin. The ergotropic effect of the hydrocolloid ration was higher than that of the conventional ration. For this reason methylated quinoxaline-1,4-dioxides are considered as belonging to the intestinal stabilisers. They increased nutrient and energy retention and decreased feed efficiency by approximately equal to 10%. Residues in the liver, the kidneys and the intestines could only be detected after 10 times the ergotropic amount.
Small groups of chickens were given feed containing either avoparcin, nitrovin, virginiamycin or zinc bacitracin from the day of their purchase as day-olds. Differences between the birds receiving growth promoters and the untreated controls were observed during the last third of the 23 d survey period. The enterococcal population of the 'dosed' birds contained a greater proportion of Enterococcus faecium than did that of the control birds while the converse was true for Ent. gallinarum. This apparent selection of Ent. faecium by the growth-promoting antibiotics had an influence on the incidence of resistance to therapeutic antibiotics among the enterococcal population as a whole. This was because this species was generally more resistant than Ent. gallinarum to cephalothin, the MLS antibiotics (erythromycin, lincomycin and tylosin) and tetracycline.
Nitrovin administered at the dose of 15 mg per 1 kg of diet, cyadox (20 mg) and virgiamycin (10 mg) were studied as to their effect on the contents of calcium, sodium, potassium and magnesium in the blood serum, breast and thigh muscles, and feathers of chickens. The contents of calcium, sodium, potassium and magnesium in the blood serum ranged within the physiological limits. An increased content of calcium and potassium and a decreased content of sodium were recorded in the breast and thigh muscles, as compared with the control group. The contents of these macroelements in the feathers of the experimental group were significantly different from those in the birds of the control group.
Chickens, fattened for 56 days, were treated with nitrovin at the dose of 15 mg per kg of feed, cyadox at the dose of 20 mg per kg of feed, and virginiamycin at the dose of 10 mg per kg of feed. After the fattening, the chickens were studied for the contents of copper, zinc, and manganese in the blood serum, thigh muscle, breast muscle, and feathers. The contents of copper, zinc and manganese in the serum ranged within the limits of physiological values. As for the breast muscle, the content of copper increased but the content of manganese declined in both breast and thigh muscle, as compared with the control group. The feathers contained more manganese in all the groups, as compared with the controls.