D. M. Storey

The Antibacterial Activity of Plant Extracts Containing Polyphenols against Streptococcus mutans

The minimum inhibitory concentrations (MIC) of commercially available and 70% aqueous propanone (P70) extracts from plants chosen for polyphenol content on Streptococcus mutans and other bacteria were determined using a standard susceptibility agar dilution technique to investigate their potential use as anticariogenic agents. The effects on adhesion of S. mutans to glass were also studied. The lowest MICs were for the P70 extracts of red grape skin (0.5 mg ml–1) and green tea and sloe berry skin (2 mg ml–1). The commercial extracts generally had a lower activity with a minimum MIC of 2 mg ml–1 for tea extracts, grape seed extracts and Pynogenol® (extract of maritime pine). All other extracts had MICs of ≧4 mg ml–1. Unfermented cocoa had greater antimicrobial activity than fermented cocoa and the activity of the fractionated extract increased with the extent of epicatechin polymerization. Epicatechin polymer had an MIC of 1 mg ml–1 and an MBC of 64 mg ml–1. Selected extracts were tested against other oral bacteria and showed activity against Gram-positive organisms. P70 extracts of unfermented cocoa, epicatechin polymer fraction, green tea and red grape seed were bacteriostatic and prevented acid production when added at the MIC to cultures of S. mutans grown in a chemically defined medium supplemented with either glucose or sucrose. There was a reduction in viability which was greater when added to washed cells, but there were some viable cells after 24 h. The extracts also reduced adherence of S. mutans to glass.

Polyols in confectionery: the route to sugar-free, reduced sugar and reduced calorie confectionery

Polyol-containing confectionery offers considerable advantages over traditional sucrose-based confectionery in terms of reduced energy content and reduced cariogenicity. However, over-consumption of polyol confectionery may lead to gastrointestinal symptoms in some individuals. Rather than consider this as a distinct disadvantage to the consumer, this article discusses how careful consideration of the physico-chemical properties of polyols and advances in product development and formulation can provide suitable polyol-based products for the consumer. Furthermore. food legislation and ingredient pricing issues are just some of the factors that must be taken into account when designing new polyol-containing products if their functional benefits and good product quality are to be delivered to the consumer.

Evaluation of solid-phase micro-extraction coupled to gas chromatography–mass spectrometry for the headspace analysis of volatile compounds in cocoa products

The aroma profile of cocoa products was investigated by headspace solid-phase micro-extraction (HS-SPME) combined with gas chromatography-mass spectrometry (GC-MS). SPME fibers coated with 100 microm polydimethylsiloxane coating (PDMS), 65 microm polydimethylsiloxane/divinylbenzene coating (PDMS-DVB), 75 microm carboxen/polydimethylsiloxane coating (CAR-PDMS) and 50/30 microm divinylbenzene/carboxen on polydimethylsiloxane on a StableFlex fiber (DVB/CAR-PDMS) were evaluated. Several extraction times and temperature conditions were also tested to achieve optimum recovery. Suspensions of the samples in distilled water or in brine (25% NaCl in distilled water) were investigated to examine their effect on the composition of the headspace. The SPME fiber coated with 50/30 microm DVB/CAR-PDMS afforded the highest extraction efficiency, particularly when the samples were extracted at 60 degrees C for 15 min under dry conditions with toluene as an internal standard. Forty-five compounds were extracted and tentatively identified, most of which have previously been reported as odor-active compounds. The method developed allows sensitive and representative analysis of cocoa products with high reproducibility. Further research is ongoing to study chocolate making processes using this method for the quantitative analysis of volatile compounds contributing to the flavor/odor profile.

Gastrointestinal tolerance of erythritol and xylitol ingested in a liquid.

Objectives:To determine and compare the gastrointestinal (GI) responses of young adults following consumption of 45 g sucrose, 20, 35 and 50 g xylitol or erythritol given as a single oral, bolus dose in a liquid.Design:The study was a randomized, double-blind, placebo-controlled study.Subjects:Seventy healthy adult volunteers aged 18–24 years were recruited from the student population of the University of Salford. Sixty-four subjects completed the study.Interventions:Subjects consumed at home without supervision and in random order, either 45 g sucrose or 20, 35 and 50 g erythritol or xylitol in water on individual test days, while maintaining their normal diet. Test days were separated by 7-day washout periods. Subjects reported the prevalence and magnitude of flatulence, borborygmi, bloating, colic, bowel movements and the passage of faeces of an abnormally watery consistency.Results:Compared with 45 g sucrose, consumption of a single oral, bolus dose of 50 g xylitol in water significantly increased the number of subjects reporting nausea (P<0.01), bloating (P<0.05), borborygmi (P<0.005), colic (P<0.05), watery faeces (P<0.05) and total bowel movement frequency (P<0.01). Also 35 g of xylitol increased significantly bowel movement frequency to pass watery faeces (P<0.05). In contrast, 50 g erythritol only significantly increased the number of subjects reporting nausea (P<0.01) and borborygmi (P<0.05). Lower doses of 20 and 35 g erythritol did not provoke a significant increase in GI symptoms. At all levels of intake, xylitol produced significantly more watery faeces than erythritol: resp. 50 g xylitol vs 35 g erythritol (P<0.001), 50 g xylitol vs 20 g erythritol (P<0.001) and 35 g xylitol vs 20 g erythritol (P<0.05).Conclusions:When consumed in water, 35 and 50 g xylitol was associated with significant intestinal symptom scores and watery faeces, compared to the sucrose control, whereas at all levels studied erythritol scored significantly less symptoms. Consumption of 20 and 35 g erythritol by healthy volunteers, in a liquid, is tolerated well, without any symptoms. At the highest level of erythritol intake (50 g), only a significant increase in borborygmi and nausea was observed, whereas xylitol intake at this level induced a significant increase in watery faeces.Sponsorship:Cerestar R&D Center, Vilvoorde, Belgium.

The host-parasite relationships in pyridoxine (vitamin B6) deficient cotton rats infected with Litomosoides carinii (Nematoda: Filaroidea)

This paper demonstrates that the establishment and growth of the filarial nematode parasite, Litomosoides carinii, is reduced in pyridoxine-deficient cotton rats. Young cotton rats were assigned to one of three dietary: vitamin B6-deficient cotton rats (B6-AL) were fed a pyridoxine-free diet ad libitum; pair-fed controls (B6 + PF) were fed the same amount of pyridoxine-free diet as animals in the deficient group and given daily oral supplements of 100 micrograms pyridoxine; and pyridoxine-sufficient controls (B6 + AL) were fed the pyridoxine-free diet ad libitum and supplemented daily with 100 micrograms pyridoxine. Half of each group was infected with 50 L3 of L. carinii by subcutaneous injection 8 weeks after the start of the experimental feeding period. B6-deficient cotton rats ate less (P < 0.001) and gained less weight (P < 0.001) than B6-supplemented controls. The levels of microfilaraemia in deficient animals, measured weekly throughout the experiment by taking blood smears, was significantly lower than in supplemented animals (P < 0.001). The deficient rats became latent for L. carinii at 20 weeks post-infection, whereas there was patent microfilaraemia in rats in the other dietary groups until the end of the experiment. Smaller (P < 0.001) and fewer (P < 0.05) adult worms were recovered from the pleural and abdominal cavities of deficient animals than from either pair-fed or sufficient controls at autopsy 28 week post-infection.

Evaluation of Chromocult coliform agar for the detection and enumeration of Enterobacteriaceae from faecal samples from healthy subjects.

The purpose of this study was to examine the use of Chromocult agar medium for isolation and enumeration of Enterobacteriaceae from human faecal samples, to compare it to MacConkey agar and to evaluate its usefulness as a possible alternative selective medium in human faecal studies. The medium was shown to be effective in identifying Escherichia coli and coliforms in faeces without the need for extensive accompanying biochemical tests for confirmation of identity. A positive correlation (r=0.86) was found between the recovery of Enterobacteriaceae on the two media, and no significant difference (P>0.05) between overall mean bacterial counts for the whole study group or at different intervals of faecal collection were observed. Chromocult agar is an effective replacement for MacConkey agar in human faecal studies and has the advantage of differentiating E. coli from other coliforms.

Vitamin A deficiency and the development of Litomosoides carinii (Nematoda, Filarioidea) in cotton rats.

SummaryThis paper demonstrates that vitamin A deficient (A−) cotton rats are more susceptible to infection withLitomosoides carinii than normal (A+) cotton rats. However, embryogenesis is frequently retarded in female worms from A− hosts.Groups of young cotton rats were depleted of their liver stores of vitamin A by feeding them a vitamin A free diet from weaning. From 6 weeks of age this diet was supplemented with either 2, 5, 10, 25 or 100 IU vitamin A per 100 g body weight per day and half the animals in each group were infected withL. carinii. At autopsy, 51 days post-infection, serum vitamin A levels were proportional to vitamin A intake but were unaffected by infection (P<0.05). A+ cotton rats grew more rapidly and gained more weight than A− ones; infected animals gained less weight than their uninfected controls. More parasites developed in rats fed either 2 or 5 IU vitamin A than those fed either 10, 25 or 100 IU (P<0.02) and parasites from the 5 IU group were significantly longer than those from all other groups (P<0.02).However, microfilarial development was retarded in female worms from A− animals as compared with A+ ones.

Gastrointestinal responses following acute and medium term intake of retrograded resistant maltodextrins, classified as type 3 resistant starch.

Design:Study part 1 was executed as a randomized double-blind placebo-controlled crossover study and study part 2 as a longitudinal study.Subjects:Forty-one healthy adult volunteers aged 18–24 years were recruited from the student population of the University of Salford. All subjects enrolled and completed study part 1 and 39 subjects enrolled and completed study part 2.Interventions:In study part 1, individuals consumed, in random order 0, 20, 40, 60, 80, 100 or 120 g of a RRM containing starch product incorporated in pre-prepared foods on individual test days. Assuming a minimum content of 50% RRM in the starch product this delivered respectively 0, 10, 20, 30, 40, 50 or 60 g of RRM. All foods were prepared and coded by personnel not involved in carrying out the tests. Test days were separated by 7 day washout periods. In study part 2, consumption of RRM was increased from 3.6 g at day 1 in incremental doses up to each subjects MNED as determined in study 1, to be achieved at day 14. Subsequently, RRM intake was from day 15–21 in a way that the final intake at day 21 was at least 10 g above the individual MNED. In both parts of the study, subjects reported the prevalence and magnitude of GI symptoms.Results:No significant change was observed in either defecation frequency and faecal consistency or the number of subjects experiencing any GI symptoms, following consumption of foods containing 0–60 g RRM. The individual MNED at which an increase in symptoms did not occur was determined as 60 g RRM for 71% of the subjects who participated in study part 1. Regression analysis showed that consumption of gradually increasing doses of RRM in food products over 21 days was associated with a significant increase in the mean symptom score for flatulence (P=1.5 × 10−4), total bowel movement frequency (P=0.023) and bowel movement frequency to pass watery faeces (P=0.0157). Increasing the ingested dose of RRM by 10 g above the predetermined MNED, however, did not provoke significant increases in GI symptoms. In both studies, the majority of symptom responses were classified by the subjects as ‘little more than usual’.Conclusions:Consumption of up to 60 g RRM is tolerated well by most individuals with no evidence of any significant dose-dependent increase in the magnitude of symptoms or the occurrence of multiple GI symptoms. However, a mild laxative effect when consuming >60 g RRM is suggested. Although there was no change in GI responses following consumption of increasing doses of RRM over 21 days, generally a dose of 10 g RRM above the MNED level was tolerated well during medium term intake.Sponsorship:Cerestar-Cargill R&D Center, Vilvoorde, Belgium.The gastrointestinal (GI) responses to ingested digestion resistant retrograded maltodextrins, having a structural similarity to type 3 resistant starch (RS) but a lower molecular weight, were studied after acute single bolus ingestion as well as during a medium-term period of daily ingestion. The overall study was split into two parts: part 1 aimed (1) to determine the GI responses of young adults following consumption of 0–60 g resistant maltodextrin; (2) to define the maximum non-effective dose (MNED) at which a considerable increase in symptoms did not occur. Part 2 aimed to determine whether a gradual increase in the daily dose of retrograded resistant maltodextrin (RRM) to a level finally exceeding by at least 10 g the individually determined MNED to acute ingestion in part 1 of the study, modified tolerance over time.

Helminth parasites of the wild rabbit Oryctolagus cuniculus near Malham Tarn, Yorkshire, UK.

Between 1992 and 1996, 95 rabbits from the immediate locality of Malham Tarn, North Yorkshire, UK were examined for the presence of helminth parasites. All the examinations took place in late September or October. Three species of nematodes, Graphidium strigosum, Passalurus ambiguus and Trichostrongylus retortaeformis and two species of cestodes, Taenia pisiformis and Cittotaenia pectinata were identified. There were no associations between helminth species richness and year of sampling, host weight or sex. A logistic model was fitted to the prevalence data from these helminths as was an over-dispersed Poisson model to the worm burden data. Graphidium strigosum was the most frequently identified species with an average prevalence of 78%. The mean prevalence and intensity of Graphidium infection were significantly effected by sampling year. The lower than normal rainfall recorded at the Tarn during the years 1995 and 1996 may have be one reason for this pattern. The worm burden of G. strigosum was significantly positively associated with rabbit body weight. The intensity of infection with P. ambiguus was significantly higher in female rabbits. There was a significant non-linear relationship between P. ambiguus worm burden and rabbit weight (P = 0.002) with worm burdens being highest in the 1000 g to 1499 g weight cohort. Trichostrongylus retortaeformis was only identified in 1994 and male rabbits harboured significantly higher worm burdens than females (48 vs. 7, P = 0.022). Over the five years, the average Taenia pisiformis prevalence was 31% and there was a significant positive association between worm burden and rabbit weight (P = 0.001). Cittotaenia pectinata had a prevalence of 37% over the whole study period with no interactions between prevalence or intensity and body weight, year of sampling or rabbit sex. All five helminths showed an overdispersed distribution with k values less than 1.

Filariasis: nutritional interactions in human and animal hosts

Vector-borne nematodes of the Order Filarioidea produce chronic, debilitating human infections which are usually nonfatal but are associated with a high degree of severe morbidity. Weight loss often accompanies infection and is probably a consequence of the increased energy cost associated with filarial fever, lymphangitis and lymphadenitis. In onchocerciasis, weight loss is associated with heavy worm loads as assessed by abundant nodules and large numbers of skin microfilariae. Experimental infections using rodent filariae have confirmed these observations and have also shown that low protein diets render hosts more susceptible to infection; nevertheless, parasite growth and embryogenesis is retarded in stunted female worms from protein deficient animals. In the absence of appropriate evidence, studies of experimental filariasis suggest that human protein-energy malnutrition may delay the development of stage-specific acquired immunity with a corresponding prolongation of patency. Epidemiological and experimental evidence shows that filarial nematodes acquire certain nutrients directly from their hosts. Of major importance in this respect is vitamin A which is taken up preferentially by human and rodent filariae; in humans, symptoms of hypovitaminosis A often accompany infection and could be an aggravating factor in onchocerciasis. Filariae also appear to require other specific nutrients such as iodine, thiamine and pyridoxine; dietary levels of these nutrients affect the host-parasite relationships in filariasis and pyridoxine seems to be of particular importance in this respect. Filarial parasites obviously compete with their hosts for available nutrients and, in the real world, human filariasis is often associated with a deterioration in the plane of nutrition of infected individuals.