Contaminated areas resulting from anthropogenic activities have, for the most part, concentrations of contaminants that exceed Tier 1 standards below which the risk is considered acceptable. However, contaminants that have been in soil for a prolonged period can become recalcitrant over time, due to various physico-chemical and biological processes. Sequestered and recalcitrant contaminants are not readily biologically available to living organisms. However, they are easily measured analytically because of the strong acid extractions that are used in the analytical methodologies. Because toxicity is a function of exposure concentration(s), exposure duration, and bioavailability, contaminants in soil can be present at concentrations that exceed established standards but they represent minimal risk to ecological receptors because the contaminants are not fully available. To predict toxicity and estimate risk, it is imperative that an accurate and reliable measure of bioavailability be available. Several surrogate measures of bioavailability were compared to the results of a battery of toxicity tests using Cu, Pb, and Zn-contaminated soils collected from a former industrial area and Cu and Zn-contaminated soils collected from a former mining site. CaCl₂extractions, hydroxypropyl-β-cyclodextrin (cyclodextrin) extractions, Simulated Earthworm Gut (SEG) tests, and bioaccumulation tests were performed using the soils. Overall, SEG-extractable Cu was most predictive of adverse effects in industrial soils, likely due to enzymatic activity and/or increased ionic strength of the solution. For the mining soils, all chemical measures of bioavailability correlated with several biological responses; however, CaCl₂-extractable Cu and SEG-extractable Cu and Zn best predicted earthworm responses. Total Cu concentrations in soil correlated best with adverse effects to plants. No method was a good predictor of all biological effects for a single organism when data from the two sites were combined. The SEG test may provide a good indication of metal toxicity at contaminated sites with varying soil physico-chemical characteristics but further validation is required.

Entrotoxigenic Escherichia coli (ETEC) K88 is a causative agent of post-weaning diarrhea (PWD) in early-weaned pigs. This study investigated the efficacy of two alternative diets, charcoal (0.1, 0.5, 1, and 2%) and a low crude protein (CP) diet (17%) supplemented with probiotic E. coli strains (UM2 and UM7), against PWD infection in ETEC K88 challenged piglets. The present study found that charcoal had no effect on the challenged piglets’ performance, ileal and colonic microbiota or their fermentation end products. There was, however, a correlation between charcoal dosage and fecal consistency score. Charcoal reduced the ileal mucosal attached ETEC K88. Feeding a low-CP diet resulted in a lower ileal ammonia concentration. The low-CP diet reduced the E. coli populations in the ileal digesta as well as lowered mRNA expression of the IL-1ß. We concluded that the use of both 1-2% charcoal diet and a low-CP diet supplemented with probiotic E. coli strains were effective in reducing the incidence and severity of PWD infection.

Vitamin D may have immunomodulatory effects in the intestine. Our objective was to determine if exposure to vitamin D mitigates intestinal inflammation in IL-10 KO mice. Mice were randomized to a diet containing 25 IU (low) or 5000 IU (high) of vitamin D/kg of diet in utero and offspring were maintained on the same diet or switched to the other diet at weaning. Fecal samples were collected at 3 months of age. Vitamin D did not affect intestinal inflammation in male and female mice and did not affect KC cytokine concentration or regulate colonic gene expression in male mice. Vitamin D modulated the gut microbiota in a sex-specific manner and depending on timing of exposure. Females in the HH group had significantly higher fecal counts of C. coccoides than the other vitamin D interventions. Therefore, vitamin D may favourably modulate microbiota composition without attenuating inflammation.

Vitamin D may have immunomodulatory effects in the intestine. Our objective was to determine if exposure to vitamin D mitigates intestinal inflammation in IL-10 KO mice. Mice were randomized to a diet containing 25 IU (low) or 5000 IU (high) of vitamin D/kg of diet in utero and offspring were maintained on the same diet or switched to the other diet at weaning. Fecal samples were collected at 3 months of age. Vitamin D did not affect intestinal inflammation in male and female mice and did not affect KC cytokine concentration or regulate colonic gene expression in male mice. Vitamin D modulated the gut microbiota in a sex-specific manner and depending on timing of exposure. Females in the HH group had significantly higher fecal counts of C. coccoides than the other vitamin D interventions. Therefore, vitamin D may favourably modulate microbiota composition without attenuating inflammation.

According to optimal foraging theory, generalist predators, such as spiders, are thought to feed indiscriminately on prey according to its availability, especially when food is scarce. In contrast, generalists can display selective feeding decisions under regimes of high prey abundance, but few studies have tracked changes in prey choice on a seasonal basis under open field conditions. Additionally, adaptations to surviving winter have been largely ignored in the research of foraging behavior. To elucidate this, I monitored prey availability and collected common forest-dwelling wolf spiders for molecular gut-content analysis, in parallel for 18 months, to assess the temporal changes occurring in spider preferences of common leaf litter prey. In addition, to determine if any physiological improvements to resisting low temperature mortality were affecting spider foraging, I also collected spiders monthly to track changes in spider supercooling points. The results revealed that spiders do exhibit selective feeding throughout the year, and appear to do so in a way that diversifies their diets. Also, despite low litter temperatures putting them in severe freezing risk, cold tolerance in these spiders remained unchanged throughout the winter, which suggests opportunity for growth during this uncompetitive period is paramount to accumulating survivorship-increasing, but also mobility-decreasing, cryoprotectants.

Entrotoxigenic Escherichia coli (ETEC) K88 is a causative agent of post-weaning diarrhea (PWD) in early-weaned pigs. This study investigated the efficacy of two alternative diets, charcoal (0.1, 0.5, 1, and 2%) and a low crude protein (CP) diet (17%) supplemented with probiotic E. coli strains (UM2 and UM7), against PWD infection in ETEC K88 challenged piglets. The present study found that charcoal had no effect on the challenged piglets’ performance, ileal and colonic microbiota or their fermentation end products. There was, however, a correlation between charcoal dosage and fecal consistency score. Charcoal reduced the ileal mucosal attached ETEC K88. Feeding a low-CP diet resulted in a lower ileal ammonia concentration. The low-CP diet reduced the E. coli populations in the ileal digesta as well as lowered mRNA expression of the IL-1ß. We concluded that the use of both 1-2% charcoal diet and a low-CP diet supplemented with probiotic E. coli strains were effective in reducing the incidence and severity of PWD infection.

Pastaraisiais dešimtmečiais industrinėse, išsivysčiusiose Vakarų Europos ir Šiaurės Amerikos šalyse ypač išaugo lėtinėmis uždegiminėmis žarnų ligomis (UŽL) sergančių vaikų skaičius. UŽL etiopatogenezė nėra pilnai aiški, tačiau manoma, kad jų išsivystymą sąlygoja pakitęs imuninis atsakas į žarnyno bakterijų antigenų stimuliaciją genetiškai predisponuotiems individams. Genetiniai tyrimai parodė genų polimorfizmo reikšmę atpažįstant bakterijas (NOD2) bei jų įtaką tokiuose ląstelės mechanizmuose, kaip imunoreguliacija (IL23R, IL10) bei autofagija (ATG16L1, IRGM). Naujausių tyrimų, atliktų su UŽL sergančiais individais, duomenimis, pakitusi žarnyno bakterijų sudėtis sukelia imuninės sistemos stimuliaciją, epitelio funkcijos sutrikimą bei gleivinės pralaidumą, dėl ko išsivysto lėtinis žarnyno gleivinės uždegimas. Remiantis klinikinių, radiologinių, endoskopinių bei histologinių tyrimų duomenimis, išskiriamos dvi UŽL: opinis kolitas (OK) ir Krono liga (KRL). OK – lėtinis storosios žarnos uždegimas, kurio metu pažeidžiami paviršiniai, t. y. gleivinės ir pogleivio, žarnos sluoksniai. KRL atveju, uždegiminiai pakitimai apima ne tik paviršinius, bet ir giliuosius žarnos sienelės sluoksnius, kurie gali atsirasti bet kurioje žarnyno dalyje, tačiau dažniausiai stebimi plonosios žarnos galinėje bei storosios žarnos pradinėje dalyse. Neseniai atlikti epidemiologiniai tyrimai parodė, kad vaikų, sergančių UŽL, sergamumas ir paplitimas kito: Krono ligos susirgimų atvejų skaičius laipsniškai... [toliau žr. visą tekstą] / In recent decades the number of children with chronic inflammatory intestinal diseases (IBD) has increased in the industrialized developed countries of Western Europe and North America [Benchimol et al., 2011; Molodecky et al., 2012]. IBD pathogenesis is not completely clear, but it is though it evolves from changes in the immune response to antigen stimulation of intestine bacteria to genetically predisposed individuals [Xavier et al., 2008]. Genetic studies have shown the importance of genetic polymorphism in identifying of bacteria (NOD2) and their influence on cell mechanisms such as immune regulation and autophagy (IRGM, ATG16L1) [Travasso et al., 2009; Cooney et al., 2009]. According to the data of the recent studies with IBD, the changed composition of the intestinal bacteria causes immune stimulation, epithelial dysfunction, and mucosal permeability, as a result, chronic inflammation of the intestinal mucosa develops. Based on the data of clinical, radiological, endoscopic and histological studies, there are two IBD subtypes: ulcerative colitis (UC) and Crohn's disease (CD). UC is a chronic inflammation of the large intestine, when surface layers, i.e. mucosa and submucosa are affected. In CD inflammatory changes include not only the surface but also the deeper layers of the intestinal wall, which can occur in any part of the intestine. These changes are often seen in the end of the small intestine and in the initial part of the large intestine. Recent... [to full text]

The FGLa/ASTs are one family of allatostatin peptides and share an amidated C-terminal sequence (FGL-amide). The inhibitory effect of FGLa/ASTs on juvenile hormone (JH) biosynthesis in Diploptera punctata led to their discovery, but there is a lack of allatostatic function across most insect species that suggests this function may not be their primary role. Rather, the FGLa/ASTs are implicated as brain/gut peptides, modulating gut physiology. This thesis demonstrates the pleiotropic nature of FGLa/ASTs in Locusta migratoria and emphasizes the role of FGLa/ASTs as brain/gut peptides involved in homeostatic processes. FGLa/AST-like immunoreactivity (FLI) is associated with the corpus cardiacum (CC) and corpus allatum (CA). FGLa/ASTs increase adipokinetic hormone release from the CC and alter JH biosynthesis from the CA, suggesting roles in energy utilization and in growth and metamorphosis. Each region of the gut exhibits FLI. The gut is dually innervated: neurons in the abdominal ganglia of the central nervous system (CNS) innervate the posterior gut and some contain FLI, while neurons within the stomatogastric nervous system (STNS) that innervate the anterior gut do not seem to contain FLI, indicating that source of FLI on the gut are cells within the CNS, which may release FGLa/ASTs at the gut to alter aspects of gut physiology. FGLa/ASTs are involved in peristalsis, neural control of foregut contractions, and ileal K+ transport. In particular, FGLa/ASTs inhibit contractions of each gut region and also modulate the rhythmic motor output of a central pattern generator within the ventricular ganglion of the STNS. FGLa/ASTs also inhibit ileal K+ efflux, suggesting a diuretic action and implicating FGLa/ASTs in fluid and ion homeostasis. This work provides a comprehensive picture of how FGLa/ASTs play an integral role in nutrient processing, energy mobilization, and growth and metamorphosis to contribute to the overall maintenance of homeostasis. This reinforces the role of FGLa/ASTs as brain/gut peptides and emphasizes their involvement in the flexibility of nervous communication and integration of the endocrine system with the CNS to achieve homeostasis.

The FGLa/ASTs are one family of allatostatin peptides and share an amidated C-terminal sequence (FGL-amide). The inhibitory effect of FGLa/ASTs on juvenile hormone (JH) biosynthesis in Diploptera punctata led to their discovery, but there is a lack of allatostatic function across most insect species that suggests this function may not be their primary role. Rather, the FGLa/ASTs are implicated as brain/gut peptides, modulating gut physiology. This thesis demonstrates the pleiotropic nature of FGLa/ASTs in Locusta migratoria and emphasizes the role of FGLa/ASTs as brain/gut peptides involved in homeostatic processes. FGLa/AST-like immunoreactivity (FLI) is associated with the corpus cardiacum (CC) and corpus allatum (CA). FGLa/ASTs increase adipokinetic hormone release from the CC and alter JH biosynthesis from the CA, suggesting roles in energy utilization and in growth and metamorphosis. Each region of the gut exhibits FLI. The gut is dually innervated: neurons in the abdominal ganglia of the central nervous system (CNS) innervate the posterior gut and some contain FLI, while neurons within the stomatogastric nervous system (STNS) that innervate the anterior gut do not seem to contain FLI, indicating that source of FLI on the gut are cells within the CNS, which may release FGLa/ASTs at the gut to alter aspects of gut physiology. FGLa/ASTs are involved in peristalsis, neural control of foregut contractions, and ileal K+ transport. In particular, FGLa/ASTs inhibit contractions of each gut region and also modulate the rhythmic motor output of a central pattern generator within the ventricular ganglion of the STNS. FGLa/ASTs also inhibit ileal K+ efflux, suggesting a diuretic action and implicating FGLa/ASTs in fluid and ion homeostasis. This work provides a comprehensive picture of how FGLa/ASTs play an integral role in nutrient processing, energy mobilization, and growth and metamorphosis to contribute to the overall maintenance of homeostasis. This reinforces the role of FGLa/ASTs as brain/gut peptides and emphasizes their involvement in the flexibility of nervous communication and integration of the endocrine system with the CNS to achieve homeostasis.

This thesis aims to assess the human health impact of exposure to estrogenic compounds from the diet. A multi-disciplinary approach is taken to address various aspects of this issue. An introduction to xenoestrogens, including international research priorities, wildlife and human health effects, mechanisms of action, structure activity relationships and additivity of estrogenic effects is provided as background information. An assessment of exposure to a range of naturally occurring and synthetic estrogenic compounds found in food is derived in Chapter 2. The assessment combines new and existing data on food concentration, food consumption and serum levels for each xenoestrogen. Exposure is combined with relative estrogenic potency data from published bioassasy data to estimate risk relative to normal circulating levels of estradiol. Assuming additivity of xenoestrogens, for an average New Zealand male and for post-menopausal women, xenoestrogens in the diet contribute an additional 12-90% of estrogenicity above normal circulating levels. For a pre-menopausal female, the contribution from the diet represents in the order of an additional 2%. The level of exposure determined in this thesis would seem to be of pharmacological relevance, especially for men with low levels of estrogen and for post-menopausal women. Bisphenol A (BPA) is an important monomer used in the manufacture of epoxy resins for internal food can linings. A survey of the BPA content of a range of 80 canned foods available to the New Zealand consumer was undertaken and the results used in the exposure and risk assessments. BPA was detected in all foods analysed except soft drinks, at concentrations ranging from <10-29 µg/kg, except for individual samples of tuna, corned beef and coconut cream that were 109, 98 and 191 µg/kg respectively. None, of over 4000 individual exposure scenarios, exceeded the temporary Tolerable Daily Intake (TDI) of 10 µg/kg body weight per day set by the Scientific Committee on Food in 2002. Intestinal microflora influence the bioavailability of the naturally occurring xenoestrogens genistein and daidzein that contribute significantly to total estrogenicity from the diet. The degradation of genistein and daidzein by the faecal microfloral of 5 human subjects was variable and unpredictable between individuals and within an individual. These findings have important implications for the promotion and prescription of soy foods and supplements for disease prevention and health benefits. The "yeast assay" is one of a number of methods available to measure estrogenicity. This assay was established and validated. In utero exposure to estrogenic compounds at critical periods of sexual differentiation and endocrine development may imprint for health effects observed later in life. Placental transfer of estrogenicity, from 17β-estradiol was studied using the human placental perfusion model and the yeast assay. The placenta provides a protective barrier to the transfer of estrogenicity. Experiments with genistein showed that 5-15% placental transfer occurred, suggesting that in utero exposure might be in the order of 10% of maternal exposure. The thesis concludes with consideration of a genomic approach to substantiate, or refute, the mechanistic link between exposure to xenoestrogens and claimed human health effect. Such an approach offers exciting opportunity to clarify the mode of action of the synthetic versus the naturally occurring xenoestrogens, to confirm or dispute additivity of effect that is an important premise of the exposure assessment, to identify key genes involved in the many possible health effects and thence risk to the individual from dietary exposure to xenoestrogens.