Indigenous enteric eosinophils control DCs to initiate a primary Th2 immune response in vivo.

Eosinophils natively inhabit the small intestine, but a functional role for them there has remained elusive. Here, we show that eosinophil-deficient mice were protected from induction of Th2-mediated peanut food allergy and anaphylaxis, and Th2 priming was restored by reconstitution with il4(+/+) or il4(-/-) eosinophils. Eosinophils controlled CD103(+) dendritic cell (DC) activation and migration from the intestine to draining lymph nodes, events necessary for Th2 priming. Eosinophil activation in vitro and in vivo led to degranulation of eosinophil peroxidase, a granule protein whose enzymatic activity promoted DC activation in mice and humans in vitro, and intestinal and extraintestinal mouse DC activation and mobilization to lymph nodes in vivo. Further, eosinophil peroxidase enhanced responses to ovalbumin seen after immunization. Thus, eosinophils can be critical contributors to the intestinal immune system, and granule-mediated shaping of DC responses can promote both intestinal and extraintestinal adaptive immunity.

Intermediate Kinematics Produce Inferior Feeding Performance in a Classic Case of Natural Hybridization

Selection on naturally occurring hybrid individuals is a key component of speciation theory, but few studies examine the functional basis of hybrid performance. We examine the functional consequences of hybridization in nature, using the freshwater sunfishes (Centrarchidae), where natural hybrids have been studied for more than a century and a half. We examined bluegill (Lepomis macrochirus), green sunfish (Lepomis cyanellus), and their naturally occurring hybrid, using prey-capture kinematics and morphology to parameterize suction-feeding simulations on divergent parental resources. Hybrid individuals exhibited kinematics intermediate between those of the two parental species. However, performance assays indicated that hybrids display performance most similar to the worse-performing species for a given parental resource. Our results show that intermediate hybrid phenotypes can be impaired by a less-than-intermediate performance and hence suffer a larger loss in fitness than could be inferred from morphology alone.

The effect of brain size evolution on feeding propensity, digestive efficiency, and juvenile growth

One key hypothesis in the study of brain size evolution is the expensive tissue hypothesis; the idea that increased investment into the brain should be compensated by decreased investment into other costly organs, for instance the gut. Although the hypothesis is supported by both comparative and experimental evidence, little is known about the potential changes in energetic requirements or digestive traits following such evolutionary shifts in brain and gut size. Organisms may meet the greater metabolic requirements of larger brains despite smaller guts via increased food intake or better digestion. But increased investment in the brain may also hamper somatic growth. To test these hypotheses we here used guppy (Poecilia reticulata) brain size selection lines with a pronounced negative association between brain and gut size and investigated feeding propensity, digestive efficiency (DE), and juvenile growth rate. We did not find any difference in feeding propensity or DE between large- and small-brained individuals. Instead, we found that large-brained females had slower growth during the first 10 weeks after birth. Our study provides experimental support that investment into larger brains at the expense of gut tissue carries costs that are not necessarily compensated by a more efficient digestive system.

Nutrient Uptake by a Deposit-Feeding Enteropneust: Nitrogenous Sources

We measured carbon, nitrogen, protein, bacterial and microalgal abundance, and mineral-specific surface area in sediments from the feeding zone of undisturbed Saccoglossus kowalewskyi, as well as in their fresh egesta. Comparison of results using surficial material 1 mm) and the top 3 mm of sediments indicated ingestion of surficial material by the enteropneusts. Assuming the surficial sediment as a food source results in apparent absorption efficiencies of 15% for TOC, 35% for TON, 60% for protein and 86% for microalgae. The C:N ratio of the apparently absorbed material was 4.2, consistent with an amino acid-rich diet. Protein- nitrogen uptake, however, accounted for only about 28% of total nitrogen absorption, indicating a dominant use of non-protein nitrogen . Bacterial and microalgal contributions to dietary nitrogen uptake were no more than 3% and 4% respectively. Active worms maintain 2 foraging areas with an average total foraging volume of 0.9 cm3 and a volume ingestion rate of 0.06 to 0.12 cm3 ind.-1 h-1. If the preferred feeding zone of these enteropneusts is the nitrogen -enriched surficial layer, we estimate that their feeding activities will deplete the available food resources every 8 to 16 h and they may rely on biological and tidal redistribution of surface material.

Behavioral and cellular analysis of classical conditioning of feeding behavior in Aplysia

In classical conditioning, an associative form of learning, animals learn to associate two stimuli. Cellular and molecular mechanisms for the induction and consolidation of associative learning and memory at the level of single cells and synaptic connections have been studied in both vertebrate and invertebrate animals. The majority of studies, however, relied on aversive stimuli to induce learning. This bias may limit the extent to which identified mechanisms generalize to other forms of associative learning and memory, such as appetitive forms. The goal of the present study was to develop a classical conditioning procedure for the marine mollusk Aplysia californica using appetitive reinforcement, and to analyze associative learning using behavioral and electrophysiological techniques. ^ Using tactile stimulation of the lips as the conditional stimulus (CS) and food as the unconditional stimulus (US) a training protocol was developed that reliably induced classical conditioning of feeding behavior. Memory persisted for at least 24 hours. The gross organization of reinforcement-mediating pathways was analyzed in additional behavioral experiments. Moreover, neurophysiological correlates of classical conditioning were identified and characterized in an in vitro preparation containing the circuitry for feeding behavior. In vitro stimulation of a nerve (AT4) that may mediate the CS during training, resulted in a greater number of buccal motor patterns (BMPs) in brains from conditioned animals, as compared to control animals. The majority of these BMPs were ingestion-like, consistent with the increased number of bites in response to the CS after classical conditioning. Moreover, classical conditioning correlated with increased excitatory synaptic input to BMP-initiating neuron B31/32, in response to stimulation of AT 4, as compared to controls. The expression of the correlates of classical conditioning identified in this study was specific to stimulation of AT 4, which is consistent the stimulus specificity that is characteristic for classical conditioning. ^ The identification of cellular correlates of classical conditioning documented here provides the basis for future, more detailed analyses of an appetitive form of associative learning and memory, that may extend the working knowledge of the cellular and molecular mechanisms for associative plasticity in general. ^

The results of a barrier analysis on complementary feeding practices among mothers in the Central River Division of the Gambia

Currently, the barriers to appropriate infant feeding practices are largely unknown in the Central River Division of the Gambia. A questionnaire was developed and implemented by a local Non Governmental Organization (NGO), the Gambia Food and Nutrition Agency, in order to gain more information and ultimately to improve the child mortality rate of the country. There were two participant groups: 88 Doers who are women who had adopted the appropriate complementary feeding practice guidelines as defined by the World Health Organization and 87 Non Doers who are women who had in some way strayed from the appropriate complementary feeding practice guidelines. The questionnaire included aspects of the Health Belief Model which can be used in the development of a future intervention. The Yes/No questions were analyzed using the Chi-square statistical method and the open-ended questions used a descriptive analysis method of evaluation. The constructs for perceived susceptibility, perceived action efficacy, perceived self efficacy, cues for action and perception of divine showed significant differences between the Doers and the Non Doers (p<0.05). The descriptive analysis revealed that both participant groups had a limited understanding of the preventative qualities of the adoption of appropriate complementary feeding practices. The women in both of groups also showed a strong perception of divine will. Women in the Central River Division perceive their husband and in-laws to be the most influential in the decision-making process regarding infant feeding practices. Recommendations for future interventions must acknowledge the importance and influence of the community surrounding the women in their adoption of the appropriate infant feeding practices. It would also be important to educate women about of the specific guidelines of the appropriate complementary feeding practices, specifically the delay in early initiation of complementary feeding. The results of this barrier analysis provide useful information to plan and implement an effective intervention to improve the child mortality rate in the Gambia. ^

A PCR based assay for the detection of enteric pathogens from Hemoccult(RTM) cards

Background. Large field studies in travelers' diarrhea (TD) in multiple destinations are limited by the need to perform stool cultures on site in a timely manner. A method for the collection, transport and storage of fecal specimens that does not require immediate processing, refrigeration and is stable for months would be advantageous. ^ Objectives. Determine if enteric pathogen bacterial DNA can be identified in cards routinely used for evaluation of fecal occult blood. ^ Methods. U.S. students traveling to Mexico in 2005-07 were followed for occurrence of diarrheal illness. When ill, students provided a stool specimen for culture and occult blood by the standard method. Cards were then stored at room temperature prior to DNA extraction. A multiplex fecal PCR was performed to identify enterotoxigenic Escherichia coli and enteroaggregative E. coli (EAEC) in DNA extracted from stools and occult blood cards. ^ Results. Significantly more EAEC cases were identified by PCR done in DNA extracted from cards (49%) or from frozen feces (40%) than by culture followed by HEp-2 adherence assays (13%). Similarly more ETEC cases were detected in card DNA (38%) than fecal DNA (30%) or culture followed by hybridization (10%). Sensitivity and specificity of the card test was 75% and 62%, respectively, and 50% and 63%, respectively, when compared to EAEC and ETEC culture, respectively, and 53% and 51%, respectively compared to EAEC multiplex fecal PCR and 56% and 70%, respectively, compared to ETEC multiplex fecal PCR. ^ Conclusions. DNA extracted from fecal cards used for detection of occult blood is of use in detecting enteric pathogens. ^

MODULATION OF HOST INTESTINAL MYOELECTRIC ACTIVITY BY LOCAL ANAPHYLAXIS: A COMPONENT OF PROTECTIVE IMMUNITY TO AN ENTERIC PARASITE (TRICHINELLA SPIRALIS, EIMERIA NIESCHULZI, BOWEL MOTILITY)

The hypothesis tested was that rapid rejection of Trichinella spiralis infective larvae from immunized rats following a challenge infection is associated with a local anaphylactic reaction, and this response should be reflected in altered small intestinal motility. The objective was to determine if altered gut smooth muscle function accompanies worm rejection based on the assumption that anaphylaxis in vivo could be detected by changes in intestinal smooth muscle contractile activity (ie. an equivalent of the Schultz-Dale reaction or in vitro anaphylaxis). The aims were to (1) characterize motility changes by monitoring intestinal myoelectric activity in conscious rats during the enteric phase of T. spiralis infection in immunized hosts, (2) detect the onset and magnitude of myoelectric changes caused by challenge infection in immunized rats, (3) determine the parasite stimulus causing changes, and (4) determine the specificity of host response to stimulation. Electrical slow wave frequency, spiking activity, normal interdigestive migrating myoelectric complexes and abnormal migrating action potential complexes were measured. Changes in myoelectric parameters induced by larvae inoculated into the duodenum of immune hosts differed from those associated with primary infection with respect to time of onset, magnitude and duration. Myoelectric changes elicited by live larvae could not be reproduced by inoculation of hosts with dead larvae, larval excretory-secretory products, or by challenge with a heterologous parasite, Eimeria nieschulzi. These results indicate that (1) local anaphylaxis is a component of the initial response to T. spiralis in immune hosts, since the rapid onset of altered smooth muscle function parallels in time the expression of rapid rejection of infective larvae, and (2) an active mucosal penetration attempt by the worm is necessary to elicit this host response. These findings provide evidence that worm rejection is a consequence of, or sequel to, an immediate hypersensitivity reaction elicited when parasites attempt to invade the gut mucosa of immunized hosts. ^

Relationship between parental feeding practices and weight status in a lower socioeconomic pre-school population

High prevalence of overweight and obesity among preschool children in the low income population is consistently documented in research with one of every seven low-income, preschool-aged children classified as obese. Parental feeding practices have the potential to be contributing factors to the obesity epidemic. However, the impact of parental feeding practices on obesity in preschool age children has not been well explored. The purpose of this study was to determine relationships between the parental feeding practices of using dessert, sweets or candy as a reward for finishing foods, restricting dessert if the child does not finish their plate at dinner, asking the child to consume everything on their plate at dinner, and having family dinners to obesity in low income, preschool age children.^ A cross-sectional secondary data analysis was completed using the STATA 11 statistical software. Descriptive statistics were completed to summarize demographic and BMI data of participants, as well as parental feeding behavior variables. Pearson’s correlation was implemented to determine a correlation between parental feeding behavior variables and BMI z scores. Predictive relationships between the variables were explored through multivariable linear regression analysis. Regression analyses were also completed factoring in the confounders of gender, age, and ethnicity.^ Results revealed (1) no significant correlations or predictive trends between the use of rewards, forced consumption, or family dinner and BMI in low income preschool age children, and (2) a significant negative correlation and predictive trend between restriction of desserts and BMI in low income preschool age children. Since the analysis supported the null hypothesis for the practices of reward use, forced consumption, and family dinner, these practices are not considered risk factors for obese level BMIs. The inverse association found for practice of restriction and BMI suggests it is unnecessary to discourage parents from using restriction. Limitations of the study included the sample size, reliability of the answers provided on the Healthy Home Survey by participant guardians, and generalizability of the sample to the larger population.^

Viability of enteric bacterial pathogens in transport media

Bacterial pathogens such as enterotoxigenic Escherichia coli, Salmonella, and Campylobacter spp. are associated with up to 80% of diarrheal illness to travelers from developed countries to developing countries. In order to study acute gastrointestinal diseases, researchers from developed countries such as the United States rely on transporting clinical specimens from the developing countries to laboratories in the U.S. in transport media systems. There are few commercially available transport media systems cited in the literature or designated by transport system manufacturers for the transport of enteric bacteria. Therefore a laboratory-based study was conducted to assess three commercial available transport media systems, two gel swabs and one liquid vial, to determine the most appropriate for the maintenance and recovery of common enteric bacterial pathogens. A total of 13 bacterial enteropathogens were recovered from 25°C and 4°C storage temperatures at time points up to 21 days. The results demonstrated that the gel swab and liquid vial transport systems performed similarly for all isolates at both temperatures. All three transport media systems struggled to maintain the isolates at recoverable concentrations when stored at 4°C and it is recommended that isolates be stored at 25°C in transport media systems. Lastly, swab transport systems are recommend for transport since they are small and easy to pack, resist leakage, and are less expensive than similarly performing liquid vial transport media systems.^

Feeding practices and their associations with diarrhea incidence in a cohort of rural Egyptian infants

A study was conducted in 4 villages in Bilbeis, Egypt, to document the infant feeding practices and identify their determinants, and examine the associations between feeding practices and diarrhea incidence in infants. A cohort of 152 infants were followed from birth with twice-weekly home visits to record feeding practices and diarrheal illness. Cross-sectional information was obtained about child birth; early neonatal feeding practices; and the socioeconomic, demographic, and water and sanitation characteristics of study families.^ Prelacteal fees were given to 60% of the infants. Nineteen percent of the infants were wet nursed at least once during the first week of life. Breast-feeding prevalence declined from 100% among infants aged less than 12 weeks to 84% among those aged 44-47 weeks. The prevalence of exclusive breast-feeding among breast-fed infants was 38% in those aged less than 4 weeks, increased to 54% in age period 4-7 weeks, and then declined rapidly to 4% in age period 24-27 weeks. The patterns and determinants of consumption by breast-fed infants of specific supplements were examined in detail.^ Between birth and age 47 weeks, the diarrhea incidence rate per person-year among breast-fed infants (6.84 episodes) was identical to the rate among all infants (6.89 episodes). In age period 0-11 weeks, the diarrhea incidence rate among breast-fed infants receiving supplements was 1.3 times (95% confidence interval: 0.9-2.0) higher than the rate among those exclusively breast-fed. In other age periods, diarrhea incidence was generally nonsignificantly higher among exclusively breast-fed infants than among those partially breast-fed and those completely weaned.^ Both univariate and multivariate analyses were done to examine the associations between diarrhea incidence and the consumption by breast-fed infants of specific supplements. After multivariate adjustment, supplements that showed significant, borderline, or suggestive positive associations with diarrhea incidence were cereal-water, cheese, raw vegetables, and 'other' foods. Significant, borderline, or suggestive negative associations were observed between diarrhea incidence and the intake of fresh animal milk, and potatoes.^ To reduce the risk of diarrhea, indiscriminate use of supplements among Bilbeis infants aged less than 12 weeks should be strongly discouraged. While mothers in this area should be educated about methods of safer preparation, handling, storage, and administration of all weaning foods, their attention should be particularly drawn to the 4 foods that were found to be positively associated with diarrhea incidence among infants in this study. ^

TRANSMISSION DYNAMICS OF ENTERIC BACTERIA IN DAY-CARE CENTERS, HOUSTON, TEXAS

An investigation was undertaken to evaluate the role of fomites in the transmission of diarrhea in day-care centers (DCC) and to elucidate the paths by which enteric organisms spread within this setting.^ During a nine-month period (December 1980-August 1981) extensive culturing of inanimate objects, as well as children and staff was done routinely each month and again repeated during diarrhea outbreaks. Air was sampled from the classrooms and toilets using a Single-Stage Sieve Sampler (Ross Industries, Midland, VA.). Stool samples were collected from both ill and well children and staff in the affected rooms only during outbreaks. Environmental samples were processed for Shigella, salmonella and fecal coliforms while stools were screened for miscellaneous enteropathogens.^ A total of 11 outbreaks occurred in the 5 DCC during the study period. Enteric pathogens were recovered in 7 (64%) of the outbreaks. Multiple pathogens were identified in 3 outbreaks. The most frequently identified pathogen in stools was Giardia lamblia which was recovered in 5 (45%) of the outbreaks. Ten of the 11 (91%) outbreaks occurred in children less than 12 months of age.^ Environmental microbiology studies together with epidemiologic information revealed that enteric organisms were transmitted from person-to-person. On routine sampling, fecal coliforms were most frequently isolated from tap handles and diaper change areas. Contamination with fetal coliforms was wide-spread during diarrhea outbreaks. Fecal coliforms were recovered with significantly greater frequency from hands, toys and other classroom objects during outbreaks than during non-outbreak period. Salmonella typhimurium was recovered from a table top during an outbreak of Salmonellosis. There was no association between the level of enteric microbial contamination in the toilet areas and the occurrence of outbreaks. No evidence was found to indicate that enteric organisms were spread by the airborne route via aerosols.^ Toys, other classroom objects and contaminated hands probably play a major role in the transmission of enteropathogens during day-care center outbreaks. The presence of many enteric agents in the environment undoubtedly explains the polymicrobial etiology of the day-care center associated diarrhea outbreaks. ^

The evidence for groundwater as a source of enteric illness in developing countries: A systematic review of the literature

Groundwater constitutes approximately 30% of freshwater globally and serves as a source of drinking water in many regions. Groundwater sources are subject to contamination with human pathogens (viruses, bacteria and protozoa) from a variety of sources that can cause diarrhea and contribute to the devastating global burden of this disease. To attempt to describe the extent of this public health concern in developing countries, a systematic review of the evidence for groundwater microbially-contaminated at its source as risk factor for enteric illness under endemic (non-outbreak) conditions in these countries was conducted. Epidemiologic studies published in English language journals between January 2000 and January 2011, and meeting certain other criteria, were selected, resulting in eleven studies reviewed. Data were extracted on microbes detected (and their concentrations if reported) and on associations measured between microbial quality of, or consumption of, groundwater and enteric illness; other relevant findings are also reported. In groundwater samples, several studies found bacterial indicators of fecal contamination (total coliforms, fecal coliforms, fecal streptococci, enterococci and E. coli), all in a wide range of concentrations. Rotavirus and a number of enteropathogenic bacteria and parasites were found in stool samples from study subjects who had consumed groundwater, but no concentrations were reported. Consumption of groundwater was associated with increased risk of diarrhea, with odds ratios ranging from 1.9 to 6.1. However, limitations of the selected studies, especially potential confounding factors, limited the conclusions that could be drawn from them. These results support the contention that microbial contamination of groundwater reservoirs—including with human enteropathogens and from a variety of sources—is a reality in developing countries. While microbially-contaminated groundwaters pose risk for diarrhea, other factors are also important, including water treatment, water storage practices, consumption of other water sources, water quantity and access to it, sanitation and hygiene, housing conditions, and socio-economic status. Further understanding of the interrelationships between, and the relative contributions to disease risk of, the various sources of microbial contamination of groundwater can guide the allocation of resources to interventions with the greatest public health benefit. Several recommendations for future research, and for practitioners and policymakers, are presented.^