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PubMed1984

Urea utilization by the intestinal flora, of infants fed mother's milk and a formula diet, as measured with the 15N-tracer technique.

15N-Incorporation by intestinal bacteria was measured under different feeding conditions in 16 infants after a single oral loading of 165 mg [15N2]urea X kg-1 body weight as a tracer. In five subjects on a mother's milk diet, the 15N-excess in the isolated intestinal bacteria was 1.08 (0.17-1.85) atom-%. The mean 15N-excess in the intestinal flora of five formula-fed subjects did not differ significantly from these values [0.63 (0.17-1.05) atom-%]. A trend to a higher incorporation of 15N from labeled urea by the intestinal flora was seen in four infants, who were adapted to an increased nutritional urea supply on a special formula, containing 14 g of milk protein, 80 g lactose, 36 g fat, and 0.35 g urea X L-1. The same observation was made in two infants with chronic renal failure. The incorporation of urea nitrogen by the putrefactive intestinal flora of infants on a formula diet as well as by the bifidobacterial flora of those on mother's milk feeding indicates the utilization of ureas as a source of bacterial protein and nucleic acid synthesis. The adaptive usage of urea for the bacterial metabolism can be considered as a sign of supportive detoxification by the intestinal flora.

Bacteria↗

PubMed2004

Postmortem production of ethanol and n-propanol in the brain of drowned persons.

We examined endogenous ethanol and n-propanol levels in the brain in 29 drowning cases in which ethanol consumption was excluded. Based on the stage of putrefaction of the brain, our cases were classified into 4 groups: pulpified brain (PB, n = 11), softened brain (SB, n = 6), discolored brain (DB, n = 2), and normal brain (NB, n = 10). The endogenous ethanol and n-propanol levels (mg/g), respectively, in the brains from these groups were 1.06 +/- 0.401 and 0.076 +/- 0.032 in PB, 0.195 +/- 0.136 and 0.012 +/- 0.009 in SB, and 0.053 +/- 0.032 and 0.001 +/- 0.001 in DB. Ethanol and n-propanol were not detected in NB. The concentration ratios of ethanol to n-propanol were 16.2 +/- 7.1 in specimens with ethanol levels > or = 0.50 mg/g (n = 10), and 17.6 +/- 13.5 in specimens with ethanol levels of 0.10 to 0.49 mg/g (n = 9). Drinking may strongly be suspected when (1) ethanol concentration in the brain is > or = 0.50 mg/g and cerebral ethanol to n-propanol ratio is > or = 40; and (2) the concentration of ethanol is 0.10 to 0.49 mg/g and the ethanol to n-propanol ratio is > or = 60.

1-Propanol↗

PubMed2005

Dumping of victims in walls.

A case is described in which a 58-year-old woman was shot and then dumped in a wall. She was immured under the staircase in the basement of a residential house by the offender, who was acting alone. The body was not found until 8 months later. The victim had been reported missing by members of her family. The cool environment delayed the onset of putrefactive changes so that it was not difficult to conduct forensic pathologic investigations into the bullet wound, with traces of gunpowder residue found on the victim's head. The way in which the offense was committed did not provide any indication of diminished responsibility on the part of the 27-year-old offender, which was reflected in the judicial consequences.

Autopsy↗

PubMed2006

Recovery of human remains after shark attack.

Two cases of fatal shark attack are reported where the only tissues recovered were fragments of lung. Case 1: An 18-year-old male who was in the sea behind a boat was observed by friends to be taken by a great white shark (Carcharodon carcharias). The shark dragged him under the water and then, with a second shark, dismembered the body. Witnesses noted a large amount of blood and unrecognizable body parts coming to the surface. The only tissues recovered despite an intensive beach and sea search were 2 fragments of lung. Case 2: A 19-year-old male was attacked by a great white shark while diving. A witness saw the shark swim away with the victim's body in its mouth. Again, despite intensive beach and sea searches, the only tissue recovered was a single piece of lung, along with pieces of wetsuit and diving equipment. These cases indicate that the only tissue to escape being consumed or lost in fatal shark attacks, where there is a significant attack with dismemberment and disruption of the integrity of the body, may be lung. The buoyancy of aerated pulmonary tissue ensures that it rises quickly to the surface, where it may be recovered by searchers soon after the attack. Aeration of the lung would be in keeping with death from trauma rather than from drowning and may be a useful marker in unwitnessed deaths to separate ante- from postmortem injury, using only relatively small amounts of tissues. Early organ recovery enhances the identification of human tissues as the extent of morphologic alterations by putrefactive processes and sea scavengers will have been minimized. DNA testing is also possible on such recovered fragments, enabling confirmation of the identity of the victim.

Adolescent↗

PubMed1976

Bifidobacteria in the intestinal tract of infants: an in-vivo study.

Weekly faecal specimens from 18 babies were examined during the first 8 weeks of life. Eight infants were breast fed, ten were bottle-fed. All suckling infants received supplementary feeds for the first 8 days. A buffer consisting of acetic acid and acetate was demonstrated in the faeces of all the breast-fed infants at some time during the period of examination. This buffer was rarely detected during the 1st week of life when supplementary feeds were given, and buffer already present gradually disappeared with the introduction of mixed feeding. In contrast, at no time was an acetate buffer demonstrated in the faeces of bottle-fed infants. Babies receiving breast milk produced faeces with low pH, high counts of saccharolytic organisms including bifidobacteria and Streptococcus faecium, and low counts of Escherichia coli, bacteroides and clostridia. Bottle-fed infants on the other hand produced faeces with a high pH and high counts of E. coli and putrefactive bacteria, but with low counts of bifidobacteria.

Acetates↗

PubMed1987

Evaluation of a monoclonal antibody-based immunoassay for detecting type A Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system.

A monoclonal antibody-based amplified enzyme-linked immunosorbent assay (ELISA) method for detecting Clostridium botulinum type A toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5-4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated, 80 degrees C/5 min + 70 degrees C/2 h, before storage at 15 degrees, 20 degrees or 27 degrees C. The presence of specific toxin was confirmed by mouse bioassay and results compared with those of the amplified ELISA method. A total of 49 strains, 39 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 95 slurry samples were tested. Fourteen of 15 strains of type A Cl. botulinum and 34 of 36 slurry samples containing type A toxin were positive by ELISA. No false positive reactions occurred with Cl. botulinum types B, C, D, E and F, or with the 10 strains of Cl. sporogenes. However, toxin produced by one strain of Cl. botulinum type A (NCTC 2012) was not detected by the amplified ELISA.

Animals↗

PubMed1988

Evaluation of a monoclonal antibody-based immunoassay for detecting type B Clostridium botulinum toxin produced in pure culture and an inoculated model cured meat system.

A monoclonal antibody-based amplified ELISA method for detecting Clostridium botulinum type B toxin was evaluated for its ability to detect the toxin in the supernatant fluid of pure cultures and after growth from Cl. botulinum spores inoculated into pork slurries. Slurries containing NaCl (1.5-4.5% w/v) and polyphosphate (0.3% w/v) were either unheated or heated 80 degrees C/5 min followed by 70 degrees C/2 h before incubation at 15 degrees, 20 degrees or 27 degrees C. Presence of specific toxin was confirmed by mouse bioassay and results were compared with those of the amplified ELISA method. A total of 48 strains, consisting of 38 Cl. botulinum and 10 Cl. sporogenes (putrefactive anaerobes), and 140 slurry samples were tested. Cultures of eight out of nine strains of type B Cl botulinum and 73 of 101 slurry samples containing type B toxin were positive by ELISA; the remaining 28 slurry samples contained type B toxin at levels below or close to the detection limit (20 LD50/ml) of the type B ELISA. No false-positive reactions occurred with Cl. botulinum types A, C, D, E or F, or with the 10 strains of Cl. sporogenes. Toxin produced by one strain of Cl. botulinum type B (NCTC 3807) was not detected by this single monoclonal antibody-based amplified ELISA. With a mixture of two monoclonal antibodies, however, the toxin from NCTC 3807 could be detected without reducing the sensitivity of the ELISA.

Animals↗

PubMed1994

Responses of the sheep blowflies Lucilia sericata and L. cuprina to odour and the development of semiochemical baits.

The literature relating to the attraction of the sheep blowflies Lucilia sericata and Lucilia cuprina to their ovine hosts is reviewed. The responses of the two species are similar and different components of the behaviour leading to host location and oviposition appear to involve at least two distinct sets of semiochemical cues. Activation, upwind orientation and landing appear to occur in response to putrefactive sulphur-rich volatiles, originating from bacterial decomposition products. Oviposition is elicited primarily by the presence of decomposition products. Oviposition is elicited primarily by the presence of ammonia-rich compounds; moisture, pheromones and tactile stimuli may also act as oviposition stimuli. There is a pronounced sex difference in the response of Lucilia to semiochemicals with a higher proportion of females attracted than males and a higher proportion of gravid than non-gravid females. While the mechanisms of host location by Lucilia are of intrinsic interest, understanding the responses to semiochemicals is important in the attempt to develop powerful synthetic baits for deployment with the traps or targets used for population sampling or suppression. The literature is discussed with respect to the development of synthetic semiochemical baits.

Animals↗

PubMed2000

Antibiotic production in relation to bacterial growth and nematode development in Photorhabdus--Heterorhabditis infected Galleria mellonella larvae.

The population of Photorhabdus luminescens C9, bacterial symbiont of the entomopathogenic nematode, Heterorhabditis megidis 90, increased rapidly to 1.2-2.6x10(9) cells g(-1) wet Galleria mellonella larvae within 24 h of nematode infection of the larvae, and maintained a relatively constant level (1.2-2.0x10(10) cells g(-1)) through the entire 14-day period of nematode development. The antibiotic, 3, 5-dihydroxy-4-isopropylstilbene, was produced by P. luminescens C9 after 24 h of nematode infection, increased rapidly at 2-5 days postinfection and remained at a level of 3000-3600 microg g(-1) wet larvae until about 21 days, decreasing gradually thereafter. The early production and continued presence of a relatively large amount of 3,5-dihydroxy-4-isopropylstilbene in the infected insect supports the hypothesis that the antibiotics produced by the bacterial symbiont help minimize competition from other microorganisms and prevents the putrefaction of the nematode-infected insect cadaver.

Animals↗

PubMed1997

Breath odor: etiopathogenesis, assessment and management.

Oral malodor is a significant social disability which usually has a local cause, particularly arising from the oral flora. Malodor may arise from the lingual or periodontal flora, gram-negative anaerobes being the main organisms capable of releasing sulphur compounds from the putrefaction of debris and other material. Volatile sulphur compounds, cadaverine and other substances appear responsible for much of the malodor. Systemic disease may underlie some cases of oral malodor. The objective assessment of malodor is still best performed by the human sense of smell (organoleptic method) but more quantifiable measures are being developed.

Bacterial Physiological Phenomena↗

PubMed2004

Insoluble zinc, cupric and tin pyrophosphates inhibit the formation of volatile sulphur compounds.

Oral malodour is mainly a result of the production of volatile sulphur compounds (VSC). The present study was concerned with investigating the anti-VSC effect of insoluble pyrophosphates (PP) of zinc, copper(II) and tin(II). The hypothesis to be tested was that the sulphide anions produced when VSC are solubilized in water have a higher affinity for the respective metal ions than the PP anion. The anti-VSC effects of insoluble PP were compared with the corresponding soluble metal salts using three in vitro methods: saliva putrefaction; dialysis of a suspension of PP and saliva against water; and analysis of water containing hydrogen sulphide and methyl mercaptan gases, and gases in the headspace. The levels of VSC were analysed by gas chromatography in the first and third methods, and released metal ions were analysed by atomic absorption spectroscopy in the second. The results showed that: the insoluble metal PP inhibited VSC formation in saliva by 99-100%; under dialysis, only minute amounts of metal ions are released from the combination of PP and saliva; and the PP lost their metal cations in water containing dissolved gases and inhibited VSC formation. Hence, the results support the experimental hypothesis. Sulphide ions are obviously very strong ligands for these metal ions.

Adult↗

PubMed1992

Volatile sulfur compounds in mouth air from clinically healthy subjects and patients with periodontal disease.

Volatile sulfur compounds (VSC) in mouth air were estimated by gas chromatography. The amount of VSC and the methyl mercaptan/hydrogen sulfide ratio were significantly increased in patients with periodontal disease. These two parameters also increased in proportion to the bleeding index and probing depth. A study was also done on the effect of removal of tongue coating on VSC concentrations in mouth air from patients with periodontal involvement. VSC and the methyl mercaptan/hydrogen sulfide ratio were reduced to 49% and 35%, respectively, by removal of the tongue coating. The average amount of tongue coating removed from patients with periodontal disease was significantly higher than from controls (90.1 mg vs. 14.6 mg, p less than 0.01). Estimated production of VSC from tongue coating was 4 times higher than the control value, and the methyl mercaptan/hydrogen sulfide ratio was also markedly increased. However, a saliva putrefaction study suggested that saliva does not contribute to the elevated ratio of methyl mercaptan in mouth air. These results strongly suggest that, in addition to periodontal pockets, tongue coating has an important role in VSC production, in particular leading to an elevated concentration of methyl mercaptan, which is more pathogenic than hydrogen sulfide.

Adult↗

PubMed1995

Influence of two infant formulas and human milk on the development of the faecal flora in newborn infants.

The establishment of the faecal flora of 39 full-term infants fed exclusively on breast milk (n = 20) or with two different modern adapted cow's milk formulas (n = 19) was studied during the first 3 months of life. One formula investigated was based on 100% bovine casein as the protein source whereas the other formula contained bovine milk proteins with a whey/casein ratio of 60:40. A faecal flora rich in bifidobacteria was found in all study groups; the growth of putrefactive bacteria (especially Bacteroides spp.), however, was limited. In formula-fed infants, significantly higher bacterial counts of enterococci and clostridia were detected compared to breast milk-fed infants. Similarities and differences due to the feeding regimen were particularly reflected in the pattern of the anaerobic bacterial species. Bifidobacterium bifidum, B. infantis and B. breve constituted the majority of the bifidobacterial flora independent of the type of milk feeding. Other bifidobacterial species such as B. longum, B. adolescentis, B. parabifidum and B. pseudo-catenulatum were detected in high numbers and at low frequencies in breastfed infants. The latter three were observed in infants fed the whey/casein formula as well. It seems that infants fed a casein formula develop a faecal flora more like that of breastfed infants concerning Lactobacillus spp. (especially L. fermentum and L. brevis).

Animals↗

PubMed1976

Bacterial fermentation of cheese whey for production of a ruminant feed supplement rich in curde protein.

A simple and efficient process for the production of a ruminant feed supplement, rich in crude protein (defined as total N X 6.25), by bacterial fermentation of cheese whey has been developed. The lactose in unpasteurized whey is fermented to lactate acid by Lactobacillus bulgaricus at a temperature of 43 degrees C and pH 5.5. The lactic acid produced is continually neutralized with ammonia to form ammonium lactate. The fermented product is concentrated by evaporation to a solids content of about 70% and adjusted to pH 6.8 with additional ammonia. The concentrated product contains about 55% crude protein. Approximately 6 to 8% of the crude protein is derived from bacterial cells. 17% from whey proteins, and 75 to 77% from ammonium lactate. The efficiency of conversion of lactose to lactic acid usually exceeds 95%. The fermentation time is greatly reduced upon the addition of 0.2% yeast extract or 0.1% corn steep liquor as a source of growth factors. Whey containing lactose at concentrations up to 7% can be fermented efficiently, but at higher concentrations lactose is fermented incompletely. The process has been scaled up to a pilot plant level, and 40 tons of concentrated product were produced fro animal feeding trials, without ever encountering putrefactive spoilage.

Animal Feed↗

PubMed1979

Separation of botulinum-positive and -negative fish samples by means of a pattern recognition method applied to headspace gas chromatograms.

A gas chromatographic headspace technique was used to analyze the gas produced during putrefaction of pond-raised, degutted trout, incubated in evacuated plastic pouches. The following samples were analyzed; 10 samples which, due to natural contamination with Clostridium botulinum, were toxic when injected into mice, 10 samples which were nontoxic when injected, and 9 samples inoculated with one strain of C. botulinum type E. The gas chromatograms showed the presence of 118 compounds in most samples. Quantitative differences among most chromatograms could be observed, but no compound was unique to any of the three groups. By means of a specific pattern recognition method, all negative samples were shown to fall into one group and were distinctly separated from the toxic samples. No differences could be observed between the two groups of inoculated and naturally contaminated trout samples. The results suggest that headspace analysis combined with pattern recognition analysis might prove to be a valuable method for screening studies of foods containing living cells of C. botulinum.

Animals↗

PubMed1980

Effect of moderately acidic pH on heat resistance of Clostridium sporogenes spores in phosphate buffer and in buffered pea puree.

The effect of pH in the range 5.0 to 7.0 on the thermal destruction of spores of Clostridium sporogenes putrefactive anaerobe 3679 was examined by three methods: a capillary tube method in which spores were suspended in phosphate buffers, a thermoresistometer method in which spores were suspended in buffered pea puree adjusted to the same set of pH values, and a thermal death time can method in which spores were again suspended in buffered pea puree. The results indicated that increasing acidity is, in general, accompanied by decreasing heat resistance, although the pH effect was more pronounced at the higher than at the lower processing temperatures. Certain pH values appear to be critical, as they produced, in all three sets of experiments, effects which would not be predicted by the overall relationship between acidity and spore heat resistance. Differences between heat resistance in phosphate buffer as compared with that in pea puree adjusted to the same pH were also noted. D-values in buffer were found to be lower than those in pea puree, except at the highest temperatures coupled with the lowest pH values. The differences between buffer D-value and pea puree D-value were found to increase with increasing pH and with decreasing temperature. On the other hand, at all pH values examined, z-values determined in buffer were somewhat higher than those determined in pea puree adjusted to the same pH.

Buffers↗

PubMed1988

Factors influencing Clostridium botulinum spore germination, outgrowth, and toxin formation in acidified media.

Clostridium botulinum type A spores were inoculated at a level of 10(7) spores per ml into sterile beef media with protein concentrations of 1, 2, 3, 4, or 6% and acidified to pH values of 2.01 to 4.75 with hydrochloric acid or 4.19 to 4.60 with citric acid. All experimental manipulations, including blending, acidification, inoculation, incubation (30 degrees C), and analyses, were conducted in an anaerobic chamber-incubator in which atmospheric oxygen levels were maintained below 2 ppm (2 microliters/liter). Under these strict anaerobic conditions (oxidation-reduction values in media ranging from -370 to -391 mV), C. botulinum spores were consistently found to germinate, grow, and produce toxin below pH 4.6. The boundary between toxic and atoxic samples in HC1-acidified beef media was mediated by titratable acidity, pH, and protein concentration. A limiting acidity was not established for the citrate-acidified samples; all blends tested (1, 2, 3, and 4% protein and titratable acidities of 0.091 to 0.453%) became toxic within 5 weeks. At the same pH and protein concentration, citric acid was less effective than HC1 in preventing the germination of C. botulinum spores. Higher levels of cell proliferation in the beef protein, as well as enhanced gas production and putrefactive degradation, indicated that beef was a better substrate than soy for C. botulinum spores under these conditions. Reducing the inoculum to 10(4) delayed but did not prevent spore outgrowth and toxin release at pH levels below 4.6.

Anaerobiosis↗

PubMed1992

Nisin dissipates the proton motive force of the obligate anaerobe Clostridium sporogenes PA 3679.

The influence of nisin on the proton motive force (delta p) generated by glucose-energized cells of the obligate putrefactive anaerobe Clostridium sporogenes PA 3679 was determined. The components of delta p, the transmembrane potential (delta psi) and the pH gradient (delta pH), were determined from the distributions of the lipophilic cation [3H]TPP+ ([3H]tetraphenylphosphonium bromide) and [14C]salicylic acid, respectively. The cells maintained a constant delta p of -111 mV, consisting of a delta pH of 0.4 to 1.0 pH units at an external pH of 5 to 7 and a delta psi of -60 to -88 mV. Nisin, carbonyl cyanide m-chlorophenylhydrazone (CCCP), and N,N'-dicyclohexylcarbodiimide (DCCD) at pH 6.0 elicited the complete release of preaccumulated [3H]tetraphenylphosphonium bromide and [14C]salicylic acid, with a concomitant depletion of delta psi and delta pH. Nisin and DCCD caused rapid drops in intracellular ATP levels from 1.2 to 0.01 and 0.06 nmol/mg of cells (dry weight), respectively. Cells exposed to nisin and DCCD lost the ability to form colonies, thus suggesting that delta psi and delta pH are necessary for cell viability. The data suggest that depletion of delta p and exhaustion of cellular ATP reserves are the basis for nisin inhibition of C. sporogenes PA 3679.

Adenosine Triphosphate↗