Bacillus Cereus Var Research Articles

Impact of the probiotic bacteria Enterococcus faecium NCIMB 10415 (SF68) and Bacillus cereus var. toyoi NCIMB 40112 on the development of serum IgG and faecal IgA of sows and their piglets

To examine the influence of two different probiotic bacteria on the humoral immune system of swine, two animal studies were carried out with sows and their litters. The sows' feed was supplemented with either Enterococcus faecium NCIMB 10415 (SF68) or Bacillus cereus var. toyoi NCIMB 40112 beginning early in pregnancy. The total IgA content in the faeces as well as the total IgG concentration in the blood of the sows was recorded before and after weaning. The same parameters were determined in the blood and faeces of the piglets. In sows, only feed supplementation with B. cereus led to a clear increase in faecal IgA. Serum IgG levels were not significantly affected by any probiotic feeding in sows. In piglets, the group that was fed B. cereus showed significantly higher faecal IgA levels shortly before weaning, whereas in the E. faecium group, a significant decrease in IgA levels was observed one week after weaning. In both probiotic fed groups the post-weaning IgG levels were significantly decreased compared to the respective control groups. We conclude that B. cereus var. toyoi feed supplementation led to an increased intestinal IgA secretion both in sows and piglets. This effect could be related to a more successful mucosal defence which in turn led to a lower level in systemic IgG production in piglets after weaning.

Opinion of the Panel on additives and products or substances used in animal feed (FEEDAP) on the safety and efficacy of the product Toyocerin® (Bacillus cereus var. Toyoi) as a feed additive for sows from service to weaning, in accordance with Regulation

Opinion of the Panel on additives and products or substances used in animal feed (FEEDAP) on the safety and efficacy of the product Toyocerin® (Bacillus cereus var. Toyoi) as a feed additive for sows from service to weaning, in accordance with Regulation

Effects of Enterococcus faecium and Bacillus cereus var. toyoi on the morphology of the intestinal mucous membrane in piglets

Eighty piglets aged 14, 28, 35 and 56 days — weaned at day 28 — were subjected to this investigation. Each age-group consisted of five animals which were fed an Enterococcus faecium NCIMB 10415 (Cylactin®) and Bacillus cereus var. toyoi (Toyocerin®) based diet. Five animals served as controls. Tissue samples were collected immediately after sacrifice at 8.30 h a.m. from duodenum, jejunum, ileum, cecum and colon to examine intestinal morphology and histochemistry. The results showed that with respect to villus height and crypt depth supplementation of probiotics in piglets feed seemed to influence the morphology and enlargement factor not at all or only to a certain extent. With respect to the number of goblet cells, the difference between probiotic fed animals and control animals was generally extremely low. The shape of the villi of the small intestinal segments greatly varied in all age groups of control and probiotic fed animals. However, this morphological variety does not depend on the mode of feeding.

Search for antibacterial and antifungal agents from selected Indian medicinal plants

A series of 61 Indian medicinal plants belonging to 33 different families used in various infectious disorders, were screened for their antimicrobial properties. Screening was carried out at 1000 and 500 μg/ml concentrations by agar dilution method against Bacillus cereus var mycoides, Bacillus pumilus, Bacillus subtilis, Bordetella bronchiseptica, Micrococcus luteus, Staphylococcus aureus, Staphylococcus epidermidis, Escherichia coli, Klebsiella pneumoniae, Pseudomonas aeruginosa, Streptococcus faecalis, Candida albicans, Aspergillus niger and Saccharomyces cerevisiae. Twenty-eight plant extracts showed activity against at least one of the test organisms used in the screening. On the basis of the results obtained, we conclude that the crude extracts of Dorema ammoniacum, Sphaeranthus indicus, Dracaena cinnabari, Mallotus philippinensis, Jatropha gossypifolia, Aristolochia indica, Lantana camara, Nardostachys jatamansi, Randia dumetorum and Cassia fistula exhibited significant antimicrobial activity and properties that support folkloric use in the treatment of some diseases as broad-spectrum antimicrobial agents. This probably explains the use of these plants by the indigenous people against a number of infections.

Effect of Saccharomyces boulardii and Bacillus cereus var. toyoi on the humoral and cellular response of mice to vaccines

The effect of Saccharomyces boulardii and Bacillus cereus var. toyoi on the humoral and cellular response of mice to the simultaneous inoculation of a tetravalent E. coli bacterin and a viable vaccine against Canine Parvovirus is reported. Thirty-six isogenic BALB-c female mice, seven weeks old, were vaccinated with 1/20th of the dose used for pigs and dogs, respectively, and randomly divided into three groups. Group 1 received non-supplemented feed, group 2 was supplemented with S. boulardii, and group 3 with Bacillus cereus var. toyoi. The humoral response was quantified through ELISAs using specific antigens, and the cellular response quantifying IL-4 and IFNg through ELISA. Means of seroconversions to the bacterin were always higher in supplemented mice than in controls, varying from 1.6–1.8 for group 2 and from 1.2–1.4 for group 3. Seroconversions against Parvovirus for group 2 were 5.2–12 times higher, and those of group 3 were 6.8–9.1 times higher than those of controls. IL-4 was produced by spleen cells of S. boulardii supplemented mice stimulated with E. coli fimbriae.

Alteration of pharmacokinetics of oxytetracycline following oral administration of Piper longum in hens

The pharmacokinetic profile of orally administered oxytetracycline (10 mg/kg body weight) was studied 7 days post oral treatment of Piper longum (15 mg equivalent/kg) in White Leghorn hens (2-2.8 kg). On the day 8, oxytetracycline (OTC) was administered orally and blood samples were collected from the wing vein in heparinised vials for plasma separation at 0 (pre-treatment), 15, 30, 60, 120, 240, 360, 480 and 600 minutes post OTC administration. Plasma OTC concentrations were determined by microbial assay technique using Bacillus cereus var. mycoides (ATCC 11778) as test organism. The plasma levels of OTC against time were adequately described by one compartment open model. The pharmacokinetic data revealed that P.longum treated animals had significantly higher area under curve (AUC), area under the first moment of plasma drug concentration-time curve (AUMC) and mean residential time (MRT). Prior treatment of P.longum significantly reduced elimination rate constant(beta)and increased elimination half life (t(1/2beta)). The total body clearance (Cl(B)) reduced by 21%whereas total duration of pharmacological effect (t(d)) increased by 29%. The treatment with P. longum reduced loading and maintenance dose by 33.3 and 39%, respectively.

Antimicrobial studies on three species of Hypericum from the Canary Islands

The antimicrobial activity of several extracts and fractions of the aerial parts of Hypericum canariense, Hypericum glandulosum and Hypericum grandifolium was investigated using the disc diffusion and broth microdilution methods against twelve reference microorganisms (eight bacterial and four fungal strains). The methanol extract and chloroform fraction of H. canariense, as well as the methanol extracts, butanol and chloroform fractions of both H. glandulosum and H. grandifolium exhibited a good antibacterial activity against four Gram-positive bacteria (Bacillus cereus var. mycoides, Micrococcus luteus, Staphylococcus aureus and Staphylococcus epidermidis) and the Gram-negative bacterium Bordetella bronchiseptica with the diameters of growth inhibition area in the range 10–25 mm and MICs values between 0.03 and 0.29 mg/ml. Neither the infusions and aqueous fractions of the species studied nor the butanol fraction of H. canariense showed any antibacterial activity against the tested microorganisms. Amongst the active extracts, the minimum inhibitory concentration (MIC) determination showed that the H. canariense chloroform fraction was the most active against M. luteus, S. aureus and S. epidermidis. No antifungal activity was seen with any of the extracts or fractions tested. The results of this study support the use of these species in Canarian traditional medicine to treat skin infections.

Aspects of the pharmacokinetics of doxycycline given to healthy and pneumonic East African dwarf goats by intramuscular injection.

The effect of experimentally induced Pasteurella haemolytica pneumonia on the pharmacokinetics of doxycycline (Doxycen Retard) administered intramuscularly was studied in seven East African dwarf goats. The study was conducted in two consecutive phases, separated by a washout period of four weeks. The experimental infection, induced by intratracheal administration of 5 ml of 10(7) to 10(9) cfu/ml of Pasteurella haemolytica, produced a temperature rise, depression and laboured breathing within 6-12 days after inoculation. The concentrations of doxycycline in the serum were determined by a quantitative microbiological assay using an agar-gel diffusion method employing Bacillus cereus var mycoides (ATCC 11778) as the test organism, with a level of detectability of approximately 0.05 micrograms/ml. The concentration-time curve of doxycycline in the serum after intramuscular injection of 20 mg/kg bodyweight of the long-acting formulation before and after experimental infection was adequately described by a one-compartment open model. The maximum serum concentrations (Cmax) of doxycycline were lower in pneumonic goats than in healthy goats (3.87 +/- 0.52 and 5.56 +/- 0.213 micrograms/ml, respectively), suggesting an increased distribution volume in the peripheral compartment. The mean +/- SEM absorption rate (ka) before infection (1.13 +/- 0.02 h-1) was smaller than the after infection (8.23 +/- 3.81 h-1), but the difference was not significant. The apparent elimination half-life (t 1/2 beta) (24.51 +/- 0.02 h) after infection was significantly increased (p < 0.05), while the corresponding rate constant (beta) was decreased (p < 0.01). The absorption half-life (t 1/2(alpha)) (0.137 +/- 0.03 h) was significantly decreased (p < 0.01) after infection. The distribution volume (Vd(beta)) was significantly increased after infection (p < 0.05). It is concluded that, although experimental infection had an effect on the disposition kinetics of doxycycline, this was not sufficiently pronounced to require alteration of the dosage during disease.

Microbiology of activated sludge treating wastewater containing Ni(II) and Cr(VI)

The species of microorganisms dominating a laboratory-scale once-through activated sludge unit treating simulated wastewater with different influent strengths (650 and 1300 mgl−1 COD) and containing 5 to 10 mgl−1 Ni(II) and 1 to 50 mgl−1 Cr(VI) were identified by standard biochemical tests. Within this framework, combinations of metals and influent COD concentrations were tried. Organisms, namely Proteus rettgeri, Acinetobacter sp., Flavobacterium lutescens, Bacillus cereus var. albolactis, Klebsiella sp., Arthrobacter simplex, and Pseudomonas mendocina, were identified at different stages of the experiments.

Some pharmacokinetic parameters of doxycycline in east African goats after intramuscular administration of a long-acting formulation.

A compartmental and non-compartmental study was carried out on five adult goats following intramuscular administration of doxycycline at 20 mg/kg bodyweight. The concentration of the drug in serum was determined by a microbiological assay employing Bacillus cereus var mycoides (ATCC 11778) as the test organism. The mean serum concentration (Cmax) and the time of maximum concentration (Tmax) were 1.87 micrograms/ml and 0.85 h, respectively. Using compartmental analysis, the plasma concentration-time curve of doxycycline best fitted a three-compartment open model with first-order absorption. A three-phase disposition of doxycycline was found, the terminal elimination half-life being approximately 40 h. The statistical moment theory was mainly used for non-compartmental analysis. The value obtained for the mean residence time (MRT) was 16.4 h. The mean values for the volume of distribution at steady state (Vdss), determined by compartmental and non-compartmental analyses, were 8.73 and 13.19 L/kg, respectively. There were no statistically significant differences when the major pharmacokinetic parameters were compared. It was concluded that the pharmacokinetic behaviour of doxycycline in goats after intramuscular administration is characterized by a three-compartment model with a slow terminal elimination phase. Based on current knowledge, this could be due to enterohepatic recycling and/or flip-flop kinetics. The study indicated that a single intramuscular administration of 20 mg/kg of doxycycline may only provide therapeutic concentrations for up to 24 h owing to slow absorption at the injection site.

Quantitative determination of oxytetracycline in honey by cylinder plate microbioassay

A cyclinder plate microbioassay is described for the presumptive detection of oxytetracycline (OTC) in honey, using Bacillus cereus var. mycoidesas the test organism. The inhibition of this organism due to naturally occurring inhibitory substances (NOIS) in honey has been reduced to &lt;0.09 8g/g OTC equivalents. In honey with low NOIS level, this procedure detected 0.04 8g OTC/g honey. Most of the NOIS effect in honey was due to its acidic pH. Confirmation of OTC was made using the OTC resistant organism, Staphylococcus epidermidis. The use of the Difco B. cereus spore suspension method proved to be more reproducible and more sensitive than the B. cereus var. mycoicles vegetative cell suspension method. The microbioassay detects OTC in honey with greater sensitivity than other methods presently used.

Physiology of Sporolactobacillus strains isolated from different habitats and the indication of in vitro antagonism against Bacillus species

In an ecological study only low numbers of Sporolactobacillus were found in habitats such as the faeces of herbivores, the rumen of cattle and the final waste water of an abattoir. Their presence in the final waste water of an abattoir indicates their possible association with food, and, more specifically, with meat. Differences were found in some physiological characteristics. One isolate (L2404) differed from the authentic Sporolactobacillus ATCC 15538 by its inability to ferment inulin, its growth in presence of 6.5% NaCl and in 0.2% tellurite, by the isomer(s) of lactic acid produced and the mol% G + G in the DNA. One Sporolactobacillus isolate (L2407) showed antagonism against Bacillus cereus, Bacillus cereus var. mycoides, Bacillus megaterium and Bacillus subtilis.

A novel Bacillus pullulanase. Its properties and application in the glucose syrups industry.

The majority of starches used in the manu-facture of glucose syrups contain 75-85% amylopectin.1) Amylopectin is a highly branched polysaccharide consisting of linear chains of 1, 4-α-linked D-glucose residues, joined together by 1, 6-α-glucosidic linkages. The branch points occur on average every 20-25 D-glucose units, so that amylopectin contains 4-5% of 1, 6-α-glucosidic linkages.2-4) The 1, 6-α-glucosidic linkages act as a kind of barrier to the action of exo-acting, saccharif ying amylases such as glucoamylases or maltogenic /3-amylases. Endo-acting α-amylases are able to by-pass the branch points, 5) but in general are not capable cf hydrolyzing the 1, 6-α-glucosidic linkage. Recent work by Kobayashi and co-workers has shown that at least one α-amylase (from Thermoactinomyces vulgaris) can hydro-lyze 1, 6-α-glucosidic linkages, in addition to 1, 4-α-glucosidic linkages.6) Glucoamylases can slowly hydrolyze 1, 6-α-glucosidic linkages in amylopectin and partially hydrolyzed amylopectin, 7) but the action of maltogenic exo-amylases ceases as a branch point is approached.8) It is therefore obvious that the efficiency of the saccharification reaction could be improved by incorporating a specific amylopectin debranchina enzyme in the system. Debranching enzyme such as isoamylase [EC 3.2.1.68, glycogen 6-glucanohydrolase] and pullulanase [EC 3.2.1.41, pullulan 6-glucanohy-drolase] have been known for many years, 9) but their use in the glucose syrups industry is far from widespread. Pullulanases from Klebsiella pneumoniae, 10, 11) Streptomyces sp., 12) and Bacillus cereus var, mycoides13) and isoamylases from Pseudomonas amyloderamosa14, 15) and Cy-tophaga sp.16) are not sufficiently thermostable to be used at 60°C. Moreover the Pseudomonas isoamylase was the only debranching enzyme sufficiently acidophilic to be used at a pH of around 4. 5.17, 18) After an extensive screening programme, our research laboratories succeeded in isolating a species of Bacillus which produced a thermostable, acidophilic pullulanase which was free from glucoamylase, β-amylase and a-amylase side activities. Some of the properties of this enzyme will now be described.

Effect of Oxytetracycline and Benzimidazole Treatments on Blight-affected Citrus Trees1

Abstract Citrus blight is a vascular wilt disease of unknown etiology. Blight-affected ‘Pineapple’ sweet orange [Citrus sinensis (L.) Osb.] trees on rough lemon (C. jambhiri Lush.) rootstock in early stages of decline were treated with benzimidazole fungicides or with oxytetracycline (OTC) by trunk injection, soil drench, or injection plus drench. The distribution and persistence of the materials were monitored by bioassay using Bacillus cereus var. mycoides for OTC and Penicillium expansum L. for benzimidazoles. Both materials were well-distributed in the canopy following trunk injections, in the root systems following soil drenches, and though most of the tree after injection plus drench treatments. Benzimidazoles and OTC persisted within the trees for several months and relatively high levels of activity were maintained for over a year by 3 applications. OTC persisted in soil for more than 6 months; whereas, benomyl disappeared from the soil in 3 months or less. Although high levels of bactericidal or fungicidal activity were maintained in the treated trees, most treated trees declined as rapidly as the untreated control trees. The injection plus drench group treated with OTC showed a slight improvement after 2 years. None of the treatments increased tree growth or resulted in increased water uptake. High levels of zinc in trunk wood, an internal symptom of blight, were unaffected by the benzimidazole treatments, but injection plus drench treatment with OTC significantly reduced zinc levels in trunk wood. Since neither OTC nor benzimidazoles completely reversed symptoms of blight, we were unable to conclusively confirm or refute proposed bacterial or fungal etiologies for citrus blight.

Disappearance Pattern of Chlortetracycline from the Liver, Bile and Tibia Bone of Growing Chicks

After feeding a diet containing 8, 000ppm equivalent of chlortetracycline (CTC) for 4 weeks and withdrawing CTC from the diet, 5 chicks each of male meat-type chicks were sacrificed at 0, 0.125, 0.25, 0.375, 1, 2, 3, 4, 6 and 8 days after the withdrawal to obtain samples of the liver, bile and left tibia bone. Contents of CTC in the liver and bile were determined microbiologically, using Bacillus cereus var. mycoides ATCC 11778, and that in the bone spectrophotofluorometrically as described by BUYSKE et al.Following Equation 4 was found suitable to describe the change in contents of CTC in the liver and bile of growing chicks, y=A1e-3.942T+A2e-0.3531T……(4)where, y is the content of CTC (ppm equivalent) and T is days after the withdrawal of dietary CTC, A1 and A2 are constants, which are 0.626 and 0.150 for the liver, respectively, and 7.67 and 2.33 for the bile, respectively. The disappearance pattern expressed by Equation 4 can be understood by two-compartment model shown in Fig. 1. The disappearance pattern of CTC from the tibia bone can be described by simple expotential curve of Equation 5, y=43.82e-0.1281T……(5)where, y is CTC content in the bone and T is as in Equation 4.Difference in disappearance patterns from the liver, bile and egg white between CTC and OTC, and between growing chicks and laying hens is discussed. It is also discussed that the bone is suspected to be the storage site in two-compartment model and that part of CTC released from the bone is deposited again in the bone making recycling shunt in two-compartment model as shown in Fig. 3. Similarity of individual error variance of CTC and OTC contents after logarithm transformation is discussed.

Residue and Disappearance of Dietary Oxytetracycline in the Blood, Muscle, Liver and Bile of Growing Chicks

Diets containing various levels from 0 to 4, 000ppm equivalent of oxytetracycline (OTC) were fed to meat-type male chicks and after 8 weeks of age dietary OTC was withdrawn. Chicks were sacrificed at 2, 4, 6 and 8 weeks of age during feeding OTC, and at 0, 3, 6 and 9 hours and 1-8 days after the withdrawal of OTC for sampling the blood, breast muscle, liver and bile. Content of OTC in the organs was determined microbiologically using Bacillus cereus var. mycoides ATCC 11778.No residue was detected in the blood, breast muscle and lived of the chicks given OTC at the level of 20ppm.Residue of OTC in the blood, breast muscle and liver on the dietary levels over 500ppm was in proportion to the dietary level of OTC. More OTC residue was found at 2 weeks of age than at older age.Disappearance pattern from the liver and bile of the chicks after the withdrawal of 4, 000ppm of OTC can be described well by the following equation, y=A1e-c1T+A2e-c2Twhere, y is residue in the organ, T is days after the withdrawal of dietary OTC, A1 and A2 are constant, c1 and c2 are constants indicating the initial and later speed of disapperance. The constants, A1, A2, c1 and c2 were estimated from the data obtained. Application on two-compartment model to explain the disappeance pattern was discussed.

Detection of Residual Antibiotics from Slaughtered Swine Carcases

Detection of residual antibiotics, such as penicillin (PC), streptomycin (SM), and oxytetracyclin (OTC) in slaughtered swine carcases was made by the standard bioassay technique, using Sarcina lutea ATCC 9341, Bacillus subtilis ATCC 6633, and Bacillus cereus var. mycoides ATCC 11778, respectively.Specimens obtained for the examination were neck muscle, liver and kidney of 29 pigs which had been treated with the above mentioned antibiotics. In addition to this field survey, an experimental study on the reliability of disk method, for the detection of penicillin incorporated in pork extract, was also carried out comparing with the standard cup method.As the results it was found that 1 (3.7%) out of 27 muscles, 6 (29.0%) of 21 livers, and 4 (20.0%) kidney contained 0.16-1.1IU/g of PC, while 3 of 21 (14.2%) liver and 2 of 21 (9.5%) kidney contained 0.15-0.75μg/g of SM. However, none of the specimen tested contained any detectable amount of OTC.As for the reliability of the disk method, it was found that almost the same sensitivity with that of the cup method could be achieved, especially at a lower concentration of the antibiotics. Therefore, it is recommeded to use disk method as a routine screenig procedure for the detection of residual penicillin in animal tissues.

Microorganisms associated with immature Prionus laticollis (Coleoptera: Cerambycidae)

Alcaligenes sp., Bacillus cereus var. mycoides, Bacillus circulans, Bacillus sp., Escherichia intermedia, Enterobacter aerogenes, Flavobacterium sp., Streptomyces sp., and 2 unidentified gram-negative rods were isolated from the digestive tracts of 24 field-collected larvae of Prionus laticollis kept in the laboratory for 1 week or more. B. cereus var. mycoides, E. aerogenes, E. intermedia, and the unidentified rods were isolated from each larva examined within 24 hr of collection. Other types of micro-organisms were not isolated on the culture media used. Attempts to culture anaerobic bacteria from 12 larvae gave no results. B. cereus var. mycoides and a species of Cephalosporium were isolated from dead pupae and Paecilomyces (= Isaria) farinosa was found on dead last stage larvae (prepupae).

Studies on Biologically Active Substances Formed by Bacilli

The products of several Bacillus strains were investigated on rabbit serum calcium decreasing, oxytocic and toad heart function promoting activities. These products were obtained from the clear supernatant fluid of the culture medium after the cells were removed by centrifugation.For the production of rabbit serum calcium decreasing substance, Bacillus subtilis K and Bacillus natto No. 8 were found to be usefull, Bacillus megaterium KM, Bacillus cereus var. mycoides and Bacillus subtilis K produced oxytocic principle. Bacillus subtilis K, Bacillus brevis and Bacillus megaterium KM also produced toad heart function promoting factor.A procedure was developed to obtain the electrophoretically homogenous rabbit serum calcium decreasing substance from culture filtrate of Bacillus subtilis K. The crude substance was obtained as isoelectric precipitate by adjusting the culture filtrate to pH 3.0. The crude substance was purified by gel filtration on a Sephadex G-75 column, isoelectric fractionation and chromatog...

Predation By Mononchoides Potohikus (Nematoda: Diplogasteridae) in Laboratory Culture

Cultures of Mononchoides potohikus have been maintained on agar plates with a bacterium (Bacillus cereus var. mycoides). Also M. potohikus has been found to be a facultative predator, preying on a wide range of nematode species. The predation rate of females is independent of the prey species, the presence of old tracks of the prey species, and the presence of a bacterial film. Predation rate is not affected by satiation but is dependent on prey density and is affected by the presence of water on the agar plate. The predation rate of the apparently non-functional males is significantly greater than that of females.