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Effects of Different Feeding Patterns on the Gut Virome of 6-Month-Old Infants.

The gut microbiome is essential for infant health, and in recent years, the impact of enteroviruses on infant health and disease has received increasing attention. The transmission of breast milk phages to the infant gastrointestinal tract contributes to the shaping of the infant gut virome, while breastfeeding regulates the colonization of the infant gut virome. In this study, we collected fecal samples from healthy infants and analyzed the distribution characteristics of infant viral communities by viral metagenomic analysis, and analyzed the differences in infant viral communities under different feeding practices. Our results indicate that the infant intestinal virome consists of phages and eukaryotic viruses. Caudovirales and Microviridae dominated the phage composition, and except for Siphoviridae, which was more predominant in the intestines of formula-fed infants, there were no significant differences in the overall abundance of other Caudovirales and Microviridae in the intestines of infants with different feeding patterns. Breastfeeding can lead to a higher diversity of infant gut viruses through vertical transmission, and a highly diverse gut virome helps maintain the maturation of the gut microbiome. This study informs the shaping of gut virome in healthy infants by breastfeeding and contributes to further research on infant gut virome characteristics and formation processes.

Humans↗

Hypolipidemic activity of 4-phenyl-5,5-dicarbethoxy-2-pyrrolidinone in rodents.

A series of pyrrolidinones have been reported to possess hypolipidemic activity in mice. The most active agent, 4-phenyl-5-5-dicarbethoxy-2-pyrrolidinone, effectively lowered both serum cholesterol and triglyceride levels at 20-30 mg/kg/d. The agent suppressed liver mitochondrial citrate exchange, phosphatidate phosphohydrolase, and sn-glycerol-3-phosphate acyl transferase activities. Lipid content of the liver, small intestine, and serum lipoprotein fractions was reduced by drug treatment, but lipid levels increased in the bile and fecal samples, suggesting the drug accelerated lipid excretion. The mode of action of the pyrrolidinone appears similar to that of the cyclic imides.

Animals↗

Human rectal mucosal gene expression after consumption of digestible and non-digestible carbohydrates.

The effect of regular consumption of the low-digestible and prebiotic isomalt versus the digestible sucrose on gene expression in rectal mucosa was examined in a randomized double-blind crossover trial. Nineteen healthy volunteers received 30 g isomalt per day or 30 g sucrose as part of a controlled diet over two 4-week test periods with a 4-week washout period in between. At the end of each test phase rectal biopsies were obtained. After RNA extraction mucosal gene expression was assayed using GeneChip microarrays. In addition, expression of cathelicidin hCap18/LL37, cellular detoxification enzymes GSTpi, UGT1A1 and CYP3A4, cyclooxygenase 2 and barrier factors MUC2 and ZO-1 were determined by real-time RT-PCR. Microbiological analyses of fecal samples revealed a shift of the gut flora towards an increase of bifidobacteria following consumption of the diet containing isomalt. Isomalt consumption did not affect rectal mucosal gene expression in microarray analyses as compared to sucrose. In addition, the expression of cathelicidin LL37, GSTpi, UGT1A1, CYP3A4, COX-2, MUC2 and ZO-1 was not changed in rectal biopsies. We conclude that gene expression of the human rectal mucosa can reliably be measured in biopsy material taken at endoscopy. Dietary intervention with the low digestible isomalt compared with the digestible sucrose did not affect gene expression in the lining rectal mucosa.

Adult↗

Comparison of two direct methods for the determination of fatty acids in infant feces.

We have validated and compared two direct methods for the determination of fatty acids in feces by capillary gas chromatography. Method I consisted of esterification of fatty acids using acetyl chloride. Method II used boron trifluoride-methanol as esterification reagent. The two methods were assayed with and without previous freeze-drying of the fecal sample. We found that the two methods could be carried out without sample freeze-drying. Precision and recovery rates were determined and the results were satisfactory. Both methods gave similar results, but Method II has certain advantages over Method I, such as speed, safety, and better recovery rates.

Acetates↗

Fecal glucocorticoids document stress in female Barbary macaques (Macaca sylvanus).

Patterns of received aggression and the endocrine response were related to an increase in fecal glucocorticoid metabolites in an intact semifree-ranging group of Barbary macaque females in order to quantify the social stress incurred over a 20-week observation period. The novel result showed that immunoreactive cortisol and 11-oxoetiocholanolone found in the feces can indeed determine the endocrine response of the adrenal gland after a social stressor. After HPLC separation of pooled fecal samples, EIA analyses using three different assays (corticosterone, cortisol, and 11-oxoetiocholanolone) to quantify immunoreactive steroids showed that the corticosterone EIA had no distinctive immunoreactive peaks. Cortisol and 11-oxoetiocholanolone immunoassays showed respectively four and two immunoreactive substances. Time series analyses revealed a behaviorally initiated increase in concentrations of cortisol and 11-oxoetiocholanolone equivalents. Furthermore, both hormone curves exhibit comparable time functions. Either antibody is very suitable for determining glucocorticoid secretion after periods of stress.

Animals↗

Excreted metabolites of gonadal steroid hormones and corticosterone in greylag geese (Anser anser) from hatching to fledging.

Steroid hormones play major roles in the organization of the phenotype and in the activation of behavior. From hatching to fledging, they are involved in growth, development, and learning. We investigated the relationship between the ontogenetic patterns of steroid hormones and the sexual and social development of greylag goslings (Anser anser). Two groups of individually marked goslings (n = 10/5) were hand-raised under near-field conditions. 17beta-OH-androgen (AM), estrogen (EM), and corticosterone (BM) immunoreactive metabolites were measured noninvasively by enzyme immunoassay of individual fecal samples. Feces were regularly sampled from hatching to fledging. All excreted steroids were found to peak at hatching and to decrease thereafter. Gonadal steroids fluctuated more than BM, which remained at low levels throughout ontogeny after a slow decrease during the first 20 days. The pattern of BM is discussed in relation to learning processes (i.e., filial imprinting) and social stress. It is suggested that high initial BM may constrain energy allocation to growth. AM increased around the age of 20 days, when the feathers start growing, and later, together with EM, at the age of 40 days. These elevated values of gonadal steroids are discussed in relation to the sensitive phase of sexual imprinting. Females show higher EM levels than males throughout ontogeny. Furthermore, the ratio of excreted estrogen to androgen (EM/AM) of females before fledging correlates with the number of hatched and fledged goslings in their first years of reproduction. In conclusion, our data suggest a role for steroid hormones in the modulation of behavioral and morphological development in the precocial greylag geese, in agreement with the organizational-activational hypothesis.

Age Factors↗

Individual and seasonal variation in fecal testosterone and cortisol levels of wild male tufted capuchin monkeys, Cebus apella nigritus.

This study tested the "challenge hypothesis" and rank-based predictions for temporal steroid production in male tufted capuchin monkeys, Cebus apella. Fecal samples (n = 209) collected from six wild males were analyzed for testosterone and cortisol concentration by enzyme immunoassay. The temporal pattern in male steroid production was compared to female sexual activity and rates of male aggression. The top-ranking adult male did not differ from other adult males in testosterone or cortisol concentration. Mean adult testosterone was significantly higher than mean subadult testosterone throughout the year. There was a clear elevation of testosterone and cortisol in both adult and subadult males during the peak of adult female sexual activity after the birth season. In fact, the magnitude of increase in testosterone was higher than predicted for a species with low male-male aggression. However, there was no difference between nonbreeding baseline testosterone levels during the birth season, and the "breeding" baseline of testosterone in males found during asynchronous female sexual activity. Of all behavioral indices examined, the distribution of female-maintained consortships was the best predictor of mean adult male testosterone concentrations. Although in many species, elevated testosterone coincides with increased male-male aggression, in the present study, the sustained high-magnitude increase in steroids during the peak of adult female sexual activity was associated with a relatively low rate of male-male intragroup aggression.

Age Factors↗

Endocrine correlates of rank, reproduction, and female-directed aggression in male Japanese macaques (Macaca fuscata).

Fecal testosterone and cortisol levels in six wild male Japanese macaques (Macaca fuscata), three of high rank and three of low, were analyzed to investigate the hormonal correlates of rank, reproduction, and female-directed aggression. The study encompassed the 6-month mating season, from October 1999 to March 2000, during which time 251 fecal samples and approximately 550 h of behavioral data were collected. Dominant males were not found to differ from subordinate males in overall rates of aggressive or copulatory behavior. Likewise, testosterone excretion, which peaked in the early part of the mating season and declined gradually thereafter, did not differ significantly by rank. High-ranking males, however, were observed to excrete significantly higher levels of cortisol than low-ranking males, suggesting that dominance may carry costs. The two hormones were found to be inversely correlated in the two most dominant males, but independent in all others. Rate of noncontact aggression was significantly correlated with testosterone, while no significant relationships were observed between testosterone and contact aggression nor any aspect of copulatory behavior. These data further support the contention that social subordinance and stress are not inexorably linked, as well as suggest that elevated glucocorticoid concentrations in high-ranking males may reflect increased metabolic costs associated with dominant male reproductive strategy.

Aggression↗

Parent-daughter relationships and social controls on fertility in female common marmosets, Callithrix jacchus.

This study examined the reproductive status of mothers and daughters to determine if parent-eldest daughter interactions would influence the daughter's fertility in the cooperatively breeding common marmoset, Callithrix jacchus. Twice weekly fecal samples were collected for hormonal analyses from seven daughters and their mothers and analyzed for cortisol, estradiol, and progesterone by EIA. Behavioral data were collected three times weekly while the daughters were under three conditions: natal family, after removing from the family and paring with a novel male, and after removing from the paired condition and placed back with the family (renatal). Under the natal condition, five of the seven daughters exhibited ovarian cycles while their mother was pregnant or displaying ovarian cycling. The five cycling daughters spent significantly more time initiating affilative behaviors with their mothers than with their fathers and showed significant changes in their behavior over the length of the ovarian cycle. However, aggression, submission, and sexual behaviors were very low for daughters in the natal phase. No differences between cortisol levels were found for a daughter and her mother. Cortisol levels showed a significant and sustained increase upon pairing within the first 20 days. All females but one cycled while paired. Six of the seven females were accepted back into the family after pairing and five of the seven females were ovulating. These results suggest that marmoset daughters are not necessarily reproductively suppressed while living with the family. Additionally, these data suggest that female marmosets that leave their family to explore mating opportunities with other groups may return to their family without reproductive or aggressive consequences.

Adrenal Glands↗

Characterization of wild lambdoid bacteriophages: detection of a wide distribution of phage immunity groups and identification of a nus-dependent, nonlambdoid phage group.

Temperate phages were isolated from fresh human fecal samples. Lambdoid phages were screened for growth on Nus+ but not Nus- bacteria. Approximately 100 independent lysogens of Nus-dependent phages were constructed and tested for immunity to superinfection by the same Nus-dependent phages. This identified 20 different phage immunity groups, 18 of which belonged to the lambdoid phage family. The DNA from the majority of these phages hybridized with a lambda DNA probe, and approximately 50% were recognized by anti-lambda antibodies. Furthermore most were inducible by UV light. Eleven phage recombinants with different immunity were obtained when a phage from each group was coinfected with lambda or its derivative lambdaBLK20. We also identified another immunity group with 48 members. None of these hybridized with either lambda or phi80 DNA probes nor were they recognized by anti-lambda serum. Most were not induced by UV light treatment, and no recombinants were obtained when crossed with either lambda or lambdaBLK20. Consequently, this group of Nus-dependent phages represent a new nonlambdoid phage family.

Animals↗

The influenza virus gene pool in a poultry market in South central china.

We surveyed influenza activity in a live poultry market in Central China for 16 months, isolating viruses from 1% of 6360 fecal samples. We obtained multiple H3N6, H9N2, H2N9, H3N3, and H4N6 isolates and single H1N1 and H3N2 isolates. Two distinct H3 molecules were identified; other hemagglutinin subtypes were phylogenetically homogeneous. The H3N6 viruses (9 genotypes) and H9N2 viruses (4 genotypes) were genetically heterogeneous, whereas the H2N9, H3N3 and H4N6 viruses had single genotypes. Thirteen representative viruses were tested for their ability to replicate in quail and chickens. All tested viruses replicated in the respiratory tract of quail. Only nine of the viruses were shed in detectable levels in infected chickens, and four of these were detected in less than 50% of infected birds. A single H4N6 isolate caused disease and systemic spread in chickens. These findings show that quail are broadly susceptible to different subtypes of influenza A virus.

Animals↗

Reverse transcription-polymerase chain reaction detection and sequence analysis of small round-structured viruses in Japan.

Between 1985 and 1995, mass outbreaks of acute gastroenteritis caused by small round-structured virus (SRSV), occurred in eight prefectures in Japan. Fecal samples from 59 patients ill during these outbreaks were recently examined in our laboratory by electron microscopy (EM) and by reverse transcription-polymerase chain reaction (RT-PCR). For RT-PCR, we prepared two sets of primers, a set corresponding to the polymerase region of open reading frame 1 (ORF-1) and a set corresponding to the capsid region of ORF-2 of Norwalk virus (NV). The SRSV nucleic acid detection rate with these primers was more than double that achieved with EM. Most samples found by EM to contain virus particles were also positive by PCR. When the two sets of primers were used separately, the virus detection rate differed depending on the primer used, suggesting that the viral strains examined were not genetically not homogeneous. We then selected nine strains of the virus, cloned their PCR products and analyzed their base sequences. The base sequences of these strains were compared with those of reference strains including prototype NV and Snow Mountain agent (SMA). This comparison yielded the following findings: (1) SRSVs that cause mass outbreaks of gastroenteritis in Japan are genetically variable; (2) SRSV strains that are genetically similar to SMA and SRSV-OTH 25/89/J(OTH25) are dominant in Japan, but strains similar to NV are also present in this country; and (3) a strain (MI1/94) which is genetically identical to Southampton virus (SHV) was detected. Detection of SRSV using sensitive RT-PCR and analysis of the sequences of the amplification products seems to provide a useful means of studying the molecular epidemiology of SRSV.

Adult↗

The incidence of cryptosporidiosis in children: a one-year prospective survey in a general hospital in Spain.

The importance of Cryptosporidium species as a cause of diarrhea in the childhood population has increased since the examination of fecal samples has been introduced in microbiological laboratories. A one-year prospective survey carried out on 699 children from rural and urban districts examined at the University Hospital in Salamanca (Spain) revealed 8 cases of cryptosporidiosis (1.1% of the children studied). Most of the cases (5 patients) were observed in the January-April period. Seven of the eight cases occurred in children younger than 3 years of age. All the children with cryptosporidiosis were immunocompetent. Diarrhea (75.0%) and vomiting (50.0%) were the most frequent clinical features of the cryptosporidiosis. In one quarter of the cases, other pathogens were associated. A comparison is made with the incidence of giardiasis, which is the commonest intestinal parasitosis in our community.

Child↗

Epidemiology and clinical significance of Blastocystis hominis in different population groups in Salamanca (Spain).

A prospective study was carried out to investigate the epidemiology and clinical significance of Blastocystis hominis in the following groups of the population of the city of Salamanca (Spain): in children attending 11 day care centres and 7 primary schools, two fecal samples were obtained from each child, and in 1231 patients attending the Clinical Hospital. A B. hominis incidence of 5.3-10.3% was found in the day care centres and an incidence rate of 13.4-19.4% was found in the primary schools. All the cases were observed in asymptomatic children. The incidence of B. hominis was greater in children older than 3 years in the day care centres and in the 10-14 year-old group in the primary schools. A heavier parasitization was observed in the boys than in the girls and in the students of schools in areas of low socio-economic level. B. hominis was identified in 40 patients attending the Clinical Hospital (3.25% of all those studied). The maximum peak of incidence was found in subjects with ages between 10 and 14 years. A follow up study was performed on 18 patients parasitized exclusively by B. hominis; 7 of these were considered to have acute gastroenteritis and one chronic gastroenteritis associated with the protozoan. No statistically significant association was observed between the number of B. hominis cells and the presence of diarrhoea. Our results show that despite the high number of asymptomatic carriers of B. hominis in the juvenile population, this protozoan may be, on other occasions, responsible for gastrointestinal symptoms.

Adult↗

Epidemiology of human rotaviruses in a maternity unit as studied by electrophoresis of genomic RNA.

An epidemiological survey of Rotavirus infections in a maternity unit was carried out between May 1983 and August 1984. Rotaviruses have been detected throughout the survey in 184 of 528 fecal samples obtained from 3 and 6 day-old newborns. Rotavirus excretion could be detected neither from the mothers nor from the medical attendants. Rotavirus RNA was analyzed by polyacrylamide gel electrophoresis. All the strains isolated exhibited the same electropherotype. This result contrasts with the genomic variability amongst Rotavirus strains usually observed in pediatric wards, and is in agreement with some other studies conducted in maternity wards; in these locations Rotavirus infection seems to have a special epidemic status as compared with other communities.

Adult↗

Aerobactin production of serotyped Escherichia coli from urinary tract infections.

Aerobactin production was examined by a bioassay in 467 Escherichia coli urinary strains from girls. All strains were of known O:K:H serotype. 139, 119 and 112 strains were isolates from pyelonephritis (Py), cystitis (Cy) and asymptomatic bacteriuria (ABU), respectively, and 97 were from fecal samples of healthy girls (FN). The incidence of aerobactin production was significantly higher among Py strains than among ABU and FN strains (P less than 0.001) and also significantly higher than among Cy strains (P less than 0.01). Aerobactin production was associated with serotype, e.g. the majority of O6:K2:H1 strains and of O16:K1:H6 were positive while e.g. the O6:K13:H1 strains were negative. There was no consistent pattern of coappearance of aerobactin and hemolysin.

Bacteriuria↗

Differentiation of ruminal and human Streptococcus bovis strains by DNA homology and 16s rRNA probes.

Streptococcus bovis is commonly present in the rumen, but strains of S. bovis have also occasionally been isolated from human blood or fecal samples. Studies were undertaken with 16s rRNA gene sequences and DNA hybridizations to define the genetic relationships between these two groups of strains. Ruminal strains were found to yield genomic DNA restriction endonuclease digest patterns different from human strains when either the 16s rRNA gene amplified from ruminal S. bovis strain JB1 or a conserved universal 23s rRNA fragment was used as probes. A DNA probe based on the V1 region of the 16s rRNA of S. bovis JB1 was found to hybridize to DNAs of other ruminal S. bovis strains (K27FF4, 21-09-6C, five new ruminal isolates, and weak hybridization was found with DNAs from S. bovis 33317 (type strain), S. equinus 9812, and six other ruminal isolates. No hybridization occurred with strains representing different major human biotypes/homology groups (43143, 43144, 27960, V1387). All ruminal S. bovis strains had a guanosine plus cytosine DNA content of 37.4-38.8 mol% and, based on DNA-DNA genomic hybridizations, could be separated into two homology groups, one of which included S. equinus 9812 and S. bovis 33317. Both ruminal groups had less than 38% DNA homology to the human strains, indicating ruminal strains are clearly two separate species distinct from the human strains.

Animals↗

A note on the experimental uptake and clearance of Candida albicans in a young captive gull (Larus sp.).

An immature gull (Larus sp.) was placed in captivity after having been kept as a pet for several weeks and allowed to roam freely. On day one of captivity, bird feces showed the presence of Candida albicans but the yeast was absent for the next 16 days. The gull was fed only yeast-free water and fish. On day 17 only, the bird was fed fish containing C. albicans which had been isolated from a wild gull. The yeast was present in all fecal samples (2-4 per day) for the next 13 days. Beginning on day 26 and irregularly thereafter the bird was fed fish containing 100-200 mg of ketoconazole. Feces continued to show the presence of C. albicans but only sporadically because the bird continued to reinfect itself, probably by contaminating the water supply via feet or feces. After protecting the water, yeast presence in feces decreased. The gull was released on day 57; feces that day were negative for C. albicans. During the experimental period the bird displayed no clinical symptoms of candidiasis. The observations indicated that one exposure to C. albicans was sufficient to establish the carrier state and that the possibility exists for shedding a potentially dangerous microorganism over a large geographical area.

Animals↗