Dissertations / Theses on the topic 'Other biological sciences not elsewhere classified'

The use of lipid profiles from the immobilised alga Selenastrum capricornuizim was investigated as a potential indicator of heavy metal pollution in freshwater environments. The toxicity of Cu", Zn 2 and Cd2t on algal growth was determined and the effective concentration inhibiting specific growth rate by 50 % (EC5 0) for each metal was found to be 124 pM Cu, 20 pM Zn and 5.7 pM Cd respectively. The Cu 24 EC50 value for immobilised cells was also shown to be 124 j.tM, suggesting that Cu exhibits similar toxic effects on growth in both free and immobilised cells. Studies of the effects of temperature and heavy metal exposure (Cu21, Zn 2 and Cd2 +) on S. caprzcornutum demonstrated that these factors altered the fatty acid and free sterol composition of free algal cells in batch culture. A shift in temperature from 25°C to 10°C led to an increase in the relative proportion of oleate and decrease in linoleate and parinate (18:4), together with a significant increase in the composition of ergostenol. Exposure to heavy metal ions led to an increase in oleate (with all three metals) and altered relative proportions of linoleate and parinate (changes being metal specific). Metal ion treatment also increased a22 desaturation of chondrillasterol. This characteristic lipid signature when S. capricornutum was exposed to heavy metal ions was significantly different from changes associated with other environmental factors. These changes in lipid composition upon heavy metal treatment were also observed during exposure of S. capricornuiwn to lower metal concentrations typically found in polluted environments. Studies of cells immobilised within alginate beads showed that gel confinement significantly affected the biochemistry and physiology of algal cells, with a reduction in growth rate and final cells numbers. Scanning electron microscopy demonstrated that growth was mainly limited to the bead periphery. Immobilisation altered the lipid of composition of cells as a consequence of alterations in membrane fluidity and membrane disruption. The Cu uptake from solution was greater in immobilised cells than free cells, thus gel confinement did not confer any protection to cells. The characteristic and significant changes within the lipid composition of free cells with Cu treatment were similarly observed in immobilised cells but were only apparent at 124 pM Cu. Thus lipid profiles, especially sterol composition of immobilised algal cells, may potentially be utilised as sensitive and novel indicators of heavy metal pollution in freshwater environments.

Despite improved knowledge and advanced treatments of high-grade gliomas, the overall survival rate of glioma patients remains low due to the recurrences and locations of the tumour. Evidence shows that the existence of a subpopulation of cells - cancer stem cells (CSCs) may be the major obstacle in treating gliomas. CD133 and nestin have been suggested as the markers of CSCs and natural stem cells. The primary focus of this study was to identify CD133+/nestin+ stem-like cells and discover their association with multidrug resistance (MDR) related genes, i.e. multiple drug resistance I (mdrl) gene and anti-apoptotic gene (bcl-2) in human glioma compared to normal brain tissues and cell lines. Glioma and normal astrocyte cell lines have been employed for CD133 isolating purposes to characterise the association with MDR related genotype and phenotype. The chemosensitivity of the isolated CD 133 population was investigated using chemosensitivity assay. Meanwhile, a serum deprivation method was established in this study to enrich and select CD 133+ CSCs in a glioma (GOS-3) cell line. As a secondary focus of this project, the possibility of immortalisation enzyme hTERT being a discriminative masker between normal and cancer brain stem cells and the transcriptional correlation between cd133 and bmi-lIc-myc oncogenes were investigated. For the first time, findings of the current study demonstrated that 1) there was an evident increase of CD133 gene expression in glioma compared to normal brain tissues where the latter expressed low levels of CD133, P-gp and Bcl-2 than glioma tissues, with an exception of nestin expression, 2) serum deprivation enriched CD133 expression and demonstrated a direct coexpression between CD133 and drug resistance in GOS-3 cells, 3) hTERT may not be a discriminative marker for normal and cancer brain stem cells, 4) although there was a strong transcriptional association between bmil and cmyc, there was an inverse transcriptional association between these genes and cd133 in serum deprived glioma cells, suggesting that bmil may not be essential for the maintenance of glioma stem cells, and 5) CD133+ glioma and normal brain cells showed a significantly high resistance towards chemotherapeutic drugs compared to the autologous CD133- cells. In conclusion, an improved understanding of molecules contributing to the maintenance of CSCs may lead to a combined treatment, targeting both CSCs and their protective MDR phenotypes leading eventually to attractive strategies for the treatment of gliomas.

In the mammal, anaesthetics are known to act via two distinct mechanisms, either increasing inhibition via GABAA receptors (eg. Na-thiopentone) or decreasing excitation via NIvIDA receptors (eg. ketamine). The aim of this thesis is to investigate the effects of both increased inhibition and decreased excitation at the synaptic level within an invertebrate model system, something which cannot readily be done in vertebrate systems. This was achieved by carrying out experiments using both the above mentioned anaesthetics on the whole animal, isolated brain and cultured neurons. In invertebrates it has been shown that GABA and Glutamate can be both excitatory and inhibitory, and injection of GABA into Lymnaea has been shown to result in behavioural changes in feeding, locomotion, escape reactions, male mating and respiration. Injection of Na-thiopentone into the whole animal was carried out in this investigation, in order to establish the anaesthetic response of the animal model to this barbiturate. The presence of gamma-aminobutyric acid (GABA) receptors has been demonstrated in a respiratory interneuron (RPeD1) both electrophysiologically and via molecular techniques, however inmiunostaining has proved negative in RPeD1 and follower cells VD2/3 (unidirectional excitatory synapse) and VD4 (mutual inhibitory synapse). This suggests that these neurons are not themselves GABAergic, although this investigation shows the responses of these neurons to bath and direct application of GABA. Na-thiopentone did not reliably anaesthetise Lymnaea upon injection into the sole of the foot, suggesting that Na-thiopentone binds to proteins within the snail, andlor has a low affininty for the GABAA receptor in Lymnaea. Other anaesthetic studies using propofol and ketaniine have also demonstrated a lack of anaesthetic response. RPeD1 hyperpolarised and became quiescent in response to the application of high concentrations of GABA (10 3-104M), however at lower doses (1O 8-1O 5M), no effect was observed (p < 0.05). Under these conditions simultaneous recordings from VD4 showed hyperpolarisation in response to the application of GABA, whereas VD2 and VD3 exhibited excitatory responses. Presynaptic picoinjection of GABA also resulted in hyperpolarisation and quiescence in RJeD1, but the simultaneous response in VD3 was not observed. Postsynaptic application of GABA directly to 'VD2, and VD4 however, resulted in responses similar to those seen in the whole brain. VD2 and 3 also receive input 2, which hyperpolarises RPeD1 and elicits an excitatory EPSP in VD2 and 3 as this is similar to the response observed in this experiment it is possible that the effects of input 2 on RPeD1 and VD2 & 3 are mediated by GABA. As RPeDI does not stain positively for GABA and hyperpolarises in response to the drug, it seems unlikely that the postsynaptie effects are due to presynaptic release of GABA. RPeD1 has been shown to form reciprocal synapses with VD4 both in vivo and in vitro. When perfused with GABA (lmJ'i4), both cells hyperpolarised reversibly. The postsynaptic response could be due to the action of GABA presynaptically inhibiting RPeDI, or directly on postsynaptic GABA receptors. However VD4 forms connections with other cells in the brain such as input 3 which may also have resulted in this inhibitory response. RPeD1 would however have received a simultaneous excitatory input from this interneuron. Attempts were made to establish the nature of the RPeD1JVD4 synapse in these experiments, but no synapses were evident. These experiments therefore confirm the presence of GABA receptors in RPeD1 and suggest theft presence in VD2, and VD4. This investigation confirms the findings of previous studies, that injection and bath perfusion of barbiturates does not lead to responses in Lymnaea comparable to that of the mammal. In addition to it's main target site, ketamine (a frequently used intravenous anaesthetic) has also been shown to act at cholinergic receptors. The effects of ketamine on learning and memory and apoptosis in the mammalian CNS are well recognised. Within the Lymnaea CNS, VD4 and LPeD1 form a unidirectional excitatory cholinergic synapse, and this was chosen to investigate the effects of ketamine on excitatory synaptic transmission, short term potentiation and synapse formation in the invertebrate animal model. Ketamine decreased synaptic transmission between VD4 and LPeD1 in a concentration dependent manner, but did not significantly affect short term synaptic plasticity (pc0.05). While neurite outgrowth remained extensive, actual sprouting was diminished by all doses of ketamine. Cells exhibited extensive veiling, which was not present in control cells. Percentage chemical synapse formation was reduced by all doses of ketamine, and in some cases inappropriate inhibitory chemical synapses were formed. Furthermore acute, clinically relevant levels of ketamine reduce excitatory cholinergic transmission between VD4 and LPeD1, but short term plasticity is unaffected. Nerve regeneration was seriously compromised, and formation of appropriate chemical synapses greatly reduced. This data has serious implications for the clinical - use of ketamine, particularly in pregnant women, children or critical care patients where nerve regeneration and synapse formation are of great importance and long term exposure common practice. In conclusion, this work supports that of other studies which have showed that invertebrates appear to be relatively insensitive to barbiturates, whereas ketamine appears to effect excitation in a manner similar to that in the mammal.

Antiulcer agents, notably inhibitors of gastric acid secretion, have been the most successful category of drugs to be discovered in recent years; and moreover, there are currently four such agents in the world list of top 25 best selling drugs. Histamine H2 antagonists have been the number one selling pharmaceutical product for more than a decade and inhibitors of the parietal cell HIC-ATPase, so called "proton pump inhibitors" (PPI), look set to continue this success. The proposed study was designed to establish the relative efficacy and mechanisms of action of three novel agents using both in vitro and in vivo models. The three compounds namely AG-1749 (Lansoprazole), PD-136450 and transforming growth factor alpha (TGF(x) were studied to evaluate their antisecretory and antiulccr activities. Lansoprazole, the second PPI to be developed for clinical use, is a non-competitive inhibitor of the H1C-ATPase and has recently been launched in a number of countries. PD-136450 is a competitive antagonist of central and peripheral cholecystokinin-B (CCK-B) receptors (gastrin receptor) and it under clinical development as an anxiolytic but which has actions on the stomach and pancreas. Anxiolytic drug is otherwise known as anti-anxiety drugs, which are used to treat anxiety disorders, like depression, panic disorders, phobias and many personality disorders. TOFu is a polypeptide growth factor, which acts at the EGF receptor and displays potent mitogenic and antisecretory activity. The initial study deals with the comparison of the three compounds with omeprazole and ranitidine in terms of their ability to inhibit acid secretion and their activity in a range of experimental ulcer models. Potency, duration of action and activity against a range of stimulants of acid secretion (histamine, pentagastrin and basal) was determined in anaesthetized rat models by establishing dose-response relationships. The compounds represent a spectrum of activities in as much as lansoprazole is a potent, long acting inhibitor, PD-136450 is an orally active but selective inhibitor, while TOFu has a very short duration and is only active after parenteral administration. In a view to find out the mechanism of action of these drugs on gastric acid secretion, isolated gastric glands from rabbits were employed as an in vitro technique using radiolabeled 14C-aminopyrine as a marker. The results show that lansoprazole was the most potent antisecretory agent compared to other two drugs. The second phase of the study deals with the activity of the three compounds against gastric ulcers induced by acid hypersecretion, indomethacin and stress. This study enabled us to assess the extent to which antisecretory activity per se compared with other actions such as wound healing (TGFa) or anxiolytic activity (PD-136450) contribute to ulcer healing. As other workers already established that prostaglandins and nitric oxide are involved in the cytoprotective activity, the present study investigated the influence of prostaglandin and nitric oxide by using indomethacin and L-NAME pretreatment on the cytoprotective activity of lansoprazole, PD-136450 and TGFcz. Moreover, the three drugs were tested for their activities on the mucus and bicarbonate production in the stomach. It was found that lansoprazole and TGFc increased the gastric mucus secretion while PD-136450 did not show any change. Moreover it was evidenced from this study that the protective activity of PD-136450 is associated with the influence of bicarbonate secretion from the pancreas. In conclusion, the results of this study have indicated that lansoprazole, PD- 136450 and TGFct are potent antisecretory and antiulcer agents which have great therapeutic importance.

Mature skeletal muscle fibres can be classified as type I, type IIa, type IIx or type IIb fibres according to the myosin heavy chain (MHC) isoform that they express. More broadly, type I fibres are classified as slow fibres and type IIa, IIx and IIb fibres as fast fibres. However, the phenotype of an adult skeletal muscle fibre is not fixed: it displays plasticity being capable of adapting to changing activity and loading levels by either transition towards a slower phenotype or transition towards a faster phenotype. Overall, the aims of these studies were to further investigate and define the signal transduction pathways implicated in the control of skeletal muscle fibre phenotype. The ability of a fast muscle to undergo a transition towards a slower phenotype in response to chronic low-frequency stimulation (CLFS) was assessed, via metabolic enzyme activity assays and NADH-TR staining, following blockade of the calcineurin signalling pathway. Metabolic enzyme assays and northern blots were employed to assess the changes in enzyme activities and MEC isoform expression levels following blockade of the calcineurin and ERK1/2 signalling pathways in primary cultures of rat myotubes. Differences in the levels of various signal transduction proteins/transcription factors between slow and fast muscle were investigated using western blotting. The nuclear translocation kinetics of NFAT and NF-κB in response to treatment with the calcium ionophore A23187 were investigated in L6 myotubes using immunocytochernistry. Calcineurin blockade using cyclosporin A failed to prevent a decrease in lactate dehydrogenase activity and an increase in NADH-TR staining intensity, both characteristics of a transition towards a slower phenotype, following CLFS of the fast rat tibialis anterior muscle. Blockade of the ERK1/2 pathway in primary cultures of rat myotubes using U0126 significantly decreased MHC Iβ mRNA levels and significantly increased MIHC IIx, MEC IIb and perinatal MHC mRNA levels. Calcineurin blockade significantly decreased MHC Iβ and embryonal mRNA levels and significantly increased MHC IIx mRNA levels. Calcineurin blockade also significantly increased the activities of lactate dehydrogenase and creatine kinase while ERK1/2 blockade significantly increased the activities of lactate dehydrogenase, creatine kinase, hexokinase, malate dehydrogenase and β-hydroxyacyl-CoA deydrogenase. ERK1/2 and NF-κB levels were found to be higher in slow muscle compared to fast muscle while calcineurin and p38α,β levels were higher in fast muscle compared to slow muscle. No nuclear translocation of NF-κB and only limited evidence for NFAT nuclear translocation was seen in L6 myotubes following treatment with A23187. Overall these studies further characterize the roles of the ERK1/2 and calcineurin pathways in the regulation of muscle phenotype suggesting that each pathway controls some, but not all, of the genes that are differentially expressed between slow and fast muscle fibres. Western blotting suggests further signal transduction protein/transcription factor targets, the functions of which may be explored in the future.

The purpose of this research is to the investigate the effectiveness of the Local Ischemic Compressiontreatment given to the patients who suffered from a cerebrovascular event, and present themusculoskeletal disorders characterized by the presence of trigger points, specifically regardingelliptical motor performance.There exist studies that demonstrate the kinematic characteristics of linear and elliptical movement inpeople with trigger points without neurological injury, however, these characteristics have not beendescribed in patients who have suffered a cerebrovascular event. Thus, this study is novel whenanalyzing the improper motor functioning in non-paretic upper limb movement control. These typesof patients have been approached from the perspective of neurological injury, and the musculoskeletalinjury that could have an impact on movement control has been neglected.This study provides new evidence on the motor performance of the non-paretic upper limb in peoplewho are suffering from musculoskeletal pain. The obtained results helped us to understand the motorfunctioning in the study sample and offers new insights based on the kinematic characteristics andpower law analysis of the movement performed.Analysis being carried out by considering the 2/3 power law, an empirical law which helps in studyingthe relationship between the tangential hand speed and the curvature of its trajectory during curvedmotion, is widely acknowledged to be an invariant of upper-limb movement. Therefore, the study isfocused on this law to interpret the effectiveness of the Local Ischemic Compression treatment givento the patients suffering with musculoskeletal injury resulting myofascial trigger points. The obtainedresults shows that power law is not much altered by the treatment which means that the ischemiccompression has only a peripherical effect (i.e., musculoskeletal system) and does not affect the motorprogramme of the human being (i.e., central nervous system). The one possible reason of having suchresults could be small sample size. Further research is suggested with large study sample.

A survey of the community of helminths in Emydura krefftii Gray in central and northern Queensland was undertaken. Then a detailed study of Sychnocotyle kholo Ferguson, Cribb and Smales, 1999, a newly described aspidogastrean parasite of E. krefftii, was conducted. The principal aim of this study was to examine the life history, fine morphology and phylogenetic relationships of this species. Specimens of Emydura krefftii were sampled from six coastal catchments from Cooktown to Bundaberg in northern and central Queensland. Concurrently data were collated on the community structure of helminths of E. krefftii. To examine the life history of S. kholo, molluscs were collected and examined for juvenile aspidogastreans from areas sampled for turtles and S. kholo eggs were collected from the faeces of collected turtles. The study of morphology of this helminth involved staining whole mounts and sections for light microscopy for gross morphology and examination of organ systems. Scanning electron microscopy was used to examine the outer surface of the worm in detail. For fine morphology of organs, transmission electron microscopy and confocal microscopy were applied. Finally characters were identified for a phylogenetic analysis and a data matrix was constructed and executed with Phylogenetic Analysis Using Parsimony (PAUP) software.

Mining for gold and copper was undertaken for nearly 100 years from 1882 at Mount Morgan in Central Queensland. Re -processing of. tailings ceased in the early 1990s and no

mining has been conducted since. The legacy of the historical mining practices is an open cut (threatening to overflow) and acid mine drainage (AMD) extending for a considerable distance along the -Dee River.

The first -18 km- of the river downstream- of- the mine, to its junction with Fletcher Creek, is severely impacted with pH consistently below 3.5. Water metal concentrations are many times higher than the water quality standards for freshwater biota. For example, the filtered mean concentrations of Al and Cu at 4 sites in this section of the river were 87.3 and 6.45 mg/L, respectively. Fish, molluscs and shrimp were absent and macroinvertebrate species richness was limited to only insects in this severely impacted region. Water quality of the river improves at the junction with Fletcher Creek and downstream the water quality is only severely impacted during periods of flow following rain events.

This study examined the biological impacts downstream of the mine including the response of biota to flows in the river. Whilst previous studies had investigated spatial variations in macroinvertebrate communities this study also examined temporal variation and response to flow. The metal content of fish and mussels from the river were determined for the first time.

Around the world, organisms are moved from one location to another, unintentionally or intentionally, through their own means of movement or by relocation by other organisms. The movements of these organisms pose risks to their new environments and the management of these risks are the focus of biosecurity. Marine ornamental organisms are a group of these relocated organisms, that are intentionally moved around the world for the purposes of being sold to hobbyists and public displays as part of the Global Marine Aquarium Trade (GMAT). The GMAT is a network of people and businesses that all play a part in the global supply chain for marine ornamental organisms. Australia’s role in this supply chain is typically that of an exporter; shipping native and endemic invertebrates and vertebrates out of the country; however, Australia also imports marine ornamental vertebrates from other countries for the Australian Marine Aquarium Trade. In the past, aspects of the Australian marine aquarium supply chain have been analysed for the purposes of the pre-border biosecurity risks, and to estimate the value of the marine aquarium trade in Australia, yet little is known about the post-border Australian marine ornamental supply chain, how the organisms move through the network of businesses and hobbyists, and the release and epidemiological biosecurity risks associated with their movements and potential release.

The behaviour of waste biomass materials, specifically bagasse* and sawdust, in deep fluidized beds was investigated. The bagasse used was dry (less than 1% moisture by mass). Sawdust was from mixed eucalyptus hardwoods with moisture contents up to 25% by mass. A series of cold flow visualisation tests were completed in a bed of 190mm diameter using graded river sand of surface mean particle diameters of 180 and 490 microns. Bagasse was added to the bed in various quantities and the ingestion and mixing phenomena observed. The influence of distributor design, cones, and draft tubes on mixing rates were investigated for use in the combustor design. Combustion Tests using Sawdust and Bagasse were completed in a Combustor of 489mm diameter with graded river sands of surface mean particle diameters of 300,490 and 530 microns. Various configurations were tested including a shallow bed of depth 130mm, deep beds of depths up to 460mm, a Reverse Circulation Bed, and Modified Spouted Beds of depths up to 740mm. Fuel feeding systems included above bed chutes, an ingestor tube, a direct bed wall screw feeder, and a pressurised screw feeder fitted to the air supply of a Modified Spouted Bed. Bagasse was not successfully fed through the screw feeder systems used. Sawdust, which has similar fluidized bed combustion characteristics to bagasse, was used in screw feeders to indicate the possible results that could be obtained from bagasse using below bed feed systems. Configurations utilising direct below bed surface screw feed, Ingestor tube feed, and pressurised screw feed to the fluidizing air were all successful in increasing the percentage of combustion occurring below the bed surface. The best results were obtained from pre-mixed air and fuel particles entering the modified spouted bed giving combustion efficiencies of up to 60% comparable to coal. Higher efficiencies would be possible with further optimisation of the design. The results of the investigation open several avenues of development including partial gasification/combustion systems and further development of the ingestor tube, reverse circulation bed and modified spouted bed concepts. The problems encountered with the combustion of lightweight, particulate biomass fuels are now reduced to finding practical methods of fuel feeding and rate control. *Bagasse is the cellulose residue from sugar cane stalks which remains after crushing.It is particulate, fibrous, tangled and irregular in size, length and aspect ratio.

The research presented here analyses and compares textual narratives generated within policy, scholarly and popular media to discern how the sexuality or intimacy needs of people categorised as ‘learning disabled’ have been historically and more recently managed in Australia and the United Kingdom. The research uses a modified critical discourse analytical approach which, in order to clarify the distinct role of power in the construction of discourses of sexuality, is mediated by the more recent ideas of progressive phronesis offered by Flyvbjerg (1998a; 2001). The analysis identifies three broad stages in the historical development of the discourses about the sexuality of learning disabled people. The first phase was prior to the 1800s, when these people were labelled non-derogatorily as ‘idiots’, and perceived as childlike, innocent and asexual. The second stage was in the 1800s when, with the emergence of scientific rationality and medicalisation, so-called idiots became medicalised and categorised as ‘feeble-minded’ or ‘moral imbeciles’. This thesis demonstrates that, during this period, an emphasis on sexual self-denial, anxieties about venereal disease and non-procreative erotic pleasure helped to inform discourses of eugenics and learning disabled people became perceived as a sexual threat to the society. This underpinned policies of sexual control through institutionalisation, gender segregation and sterilisation. The third period in the development of discourses relating to the sexuality of learning disabled people paralleled the ‘sexual revolution’ of the late- 1900s and the move towards deinstitutionalisation and human rights. This analysis shows that, although the principle of ‘sexual freedom’ was ostensibly incorporated in modern policy discourse, the sexuality of learning disabled people continues to be influenced by significant barriers of sexual intolerance, demonstrated by continuing practices of sexual segregation, sterilisation, criminal labelling and imprisonment. The analysis indicates that a discourse of sexuality, which has legitimised the control and management of learning disabled people in varying forms since the Enlightenment, continues to be encountered in policy and popular narratives. Robust sexuality awareness and education programs for carers of these people, and society in general, are necessary so that intimacy and/or sexual desires are accepted as a normal need for all human beings.

The poultry industry has rapidly grown in the last few years with a focus in improving growth and productivity of broiler chickens, with performance assessed on measures such as feed conversion ratio, body weight gain and good immunity. The incorporation of antibiotics and feed additives in poultry diets, have been implemented for years to ensure the maintenance of poultry health with a focus on the control and reduction of zoonotic pathogens. In the last few years, however, key issues surrounding the antimicrobial resistance of antibiotics have urged for alternative supplementations. Nanoparticles (NPs) of silver and other metals have been heavily used in the poultry industry to improve the growth and performance of birds. Whilst successful, metal NPs exhibited higher toxicity at the higher surface to volume ratio, especially with the use of silver. This study proposes the use of NPs of essential metals and natural compounds to safely deliver nutrients, resulting in positive impacts on health and productivity with little to no toxic effects. Selenium is an essential mineral, required for the proper functioning of the immunity and is an important element in the first cell line of defence in the body. The work described in this thesis explores the ability of selenium NPs to improve the health and growth of broiler chickens by modulating their gut microbiome and metabolome, without the toxic effects observed with silver. Selenium NPs were synthesised using a simple chemical reduction method and a full characterisation was performed, assessing the physicochemical properties of the NP. Selenium NPs were then compared in an animal trial against two commonly used selenium additives in the poultry industry, sodium selenite (inorganic selenium) and selenomethionine (organic selenium). The performance of the birds was assessed based on body weight gain, the gut microbial composition and metabolite production. The toxicity of NPs was further investigated by quantifying selenium concentration in various tissues, along with a detailed histopathological assessment. Results show selenium NPs completely altered the gut microbial ecology at high concentration, with a strong correlation observed between Faecalibacterium prausnitzii abundance and increasing concentration of selenium NPs. Selenium NPs additionally increased villus height/crypt ratio associated with enhanced absorption in the small intestine and an overall increase of healthy colonic metabolites. Finally, an in vitro study demonstrated the ability of selenium NPs to reduce emerging pathogenic Enterococcus cecorum species. This thesis demonstrates the prospective ability of selenium NPs as alternatives to antibiotics and bulk supplementation, resulting in an improvement of health and performance of broiler chickens in the poultry industry.

New synthetic methods are explored for the synthesis of protoberberines using carbanionimine (CAI) cycloaddition, aza Diels-Alder (DA) cycloaddition.. Protoberberine alkaloids are widely distributed in numerous plant families, and commonly found as active components in many folklore medicines in Asia, Europe and North America. Due to their broad diversity of biological activities, there are great efforts about chemical, biogenetic and enzymatic synthesis, transformation, pharmacology, and culture development studies of these alkaloids. In this thesis, new synthetic methods are explored for the synthesis of protoberberines using the following approaches a) carbanionimine (CAI) cycloaddition, b) aza Diels-Alder (DA) cycloaddition. The structure elucidation of products using various analytical techniques, such as IR, UV-vis., MS, NMR and X-ray, are broadly discussed. Detailed stereostructure and conformation analysis have been studied using various one and two dimensional NMR techniques, which show great advantage for structure analysis in the liquid state, and give the consistent conclusion as the solid state structure as determined by the single crystal X-ray diffraction method. The carbanion-cyclic imine approach has been most successful in the synthesis of protoberberines, which contain a four-ring system (ABCD). Cycloaddition of 3,4-dihydroisoquinoline (DHIQ, eg A-I) with phthalide anion (eg A-2) gives stereo and regia selective 13-hydroxy protoberberines (eg A-3) with single diastereomers observed and isolated, which are transformed to various functionalised derivatives. The novel reaction of 3-methoxyphthalide anion (A-4) with DHIQ, where a 2+1 adduct (A-6) is formed, is broadly studied. The mechanisms are studied in details and highly suggestive via tandem nucleophilic condensation of the anion, in which the 8,13-dioxo intermediate (eg A-5) plays an important role. The geometry change due to the replace of angular 13a-H with methyl group accounts for the different facial selectivity of 13-keto to nucleophiles in A-5a A-5b. These are supported by theoretical AMl calculation and Surface Probe Analysis on model compounds (A-8). The phthalide anion-acyclic imine approach has been used to provide open chain products (eg A-10 and A-11). NMR and AMl calculation prove trans stereochemistry of major isomers (eg A-10). The conclusion is in contrast to previous assignments by MacLean, Cushman, and Prager on similar compounds. The direct asymmetric synthesis is also explored by using a chiral o-toluamide anion (eg A-12) instead of a phthalide counterpart. A series of chiral amine auxiliary agents have been studied and the tertiary o-toluamide with a bulky cyclohexyl group in addition to the chiral substituent on the amide nitrogen atom (A-12) gives the highest enantioselectivity (>96% ee). This approach established a new, fast entry to both enantiomers of protoberberines and exhibits predictable regiochemistry. This is an obvious advantage in all anion-imine cycloaddition reactions in our study and others. DA methodology is also studied, however only the reaction between DHIQ (AB synthon) and phthalazine (A-14, CD synthon) works. This reaction is the first example of aza DA reaction with phthalazine as an aza diene with an aza dienophile. The other AB+CD approaches with DHIQ (AB) as dienophile allowed to react with different dienes (CD), such as isobenzofuran and cyclopentadienone, do not give any adducts under thermal, Lewis acid catalyst or high pressure conditions. While the first step of an AB+C+D approach which involves sequential DA reaction of DHIQ with s-tetrazine (A-16) was successful, the second step involving benzyne or cyclic enamine cycloaddition failed.

Utilizing environmental DNA (eDNA) for the detection of species in the field is a potentially cost-effective and time-saving technology that may be useful in understanding the distribution and abundance of threatened or endangered species such as the Eastern Massasauga (Sistrurus catenatus). I describe the development of an eDNA assay for the species and evaluate its ability to detect eDNA in laboratory and field conditions. In the field samples, I also investigated the potential for abiotic conditions to influence eDNA detection. Species-specific primers and probe were designed to amplify a 152 bp segment of the massasauga COI gene. Target eDNA could be detected in samples containing as little as 100 copies of target DNA/μL. Water samples collected from laboratory housed snakes indicated that eDNA can be detected in water 56 days after massasauga removal. Field samples were taken from crayfish burrows, known overwintering habitat for the species, from four sites that vary in snake use as ascertained by traditional visual surveys. Of the 60 burrows sampled, seven had a positive detection for massasauga eDNA with no difference in detection rate between DNA extracted from burrow water and burrow sediment. Occupancy models fitted to burrow water indicated that larger amounts of total DNA in a sample may increase the probability of detection of a massasauga eDNA. Large confidence intervals in site occupancy (ѱ) and burrow detection (Θ) values suggest that a larger sample size is needed for more reliable occupancy models. Abiotic conditions within crayfish burrows varied among sites but correlation with eDNA detection was not supported. Estimates of qPCR detection within a burrow with eDNA (ρ) suggest that up to 10 qPCR replicates per burrow sample may be necessary. Further studies need to examine eDNA degradation in the field, improve upon the limit of detection, and sample a larger number of sites for eDNA sampling to be a stand-alone survey method for Eastern Massasaugas.

Environmental DNA (eDNA) is being used increasingly for biomonitoring of communities (e.g., microbes, macroinvertebrates, fish species) across terrestrial and aquatic ecosystems. Developing methods that combine eDNA approaches with metagenomic barcoded amplicon sequencing (eDNA-metabarcoding) are now providing a powerful noninvasive and cost-effective means for comprehensively surveying biodiversity in a wide range of habitats. Invasive species have a substantial impact on the ecology and economics of the Great Lakes region, and eDNAmetabarcoding methods have recently been applied in monitoring non-native, as well as native, fish populations in the freshwater systems there. In this research, we validated an eDNAmetabarcoding approach that uses established platforms, the MiFish/MitoFish pipeline, for fish community monitoring on Lake Michigan. For validation, we compared survey results from our eDNA-metabarcoding approach to those obtained using traditional surveys (e.g., electrofishing and seining). We also sampled a closed 180,000-gallon freshwater fish tank system to see how well our methods characterized a known native fish population that resided in the tank. Finally, we applied the approach to monitoring invasive and native fish populations in southern Lake Michigan at a site that is currently undergoing restoration to improve the aquatic habitats.. We were able to reliably capture the fish community structure of the native fish tank as well as those of open waters on the lake using our methods. Diversity patterns detected at the restoration site using our eDNA-metabarcoding approach accurately reflected those of the historical record, which have taken many years to establish by conventional means. Overall, this study suggests eDNAmetabarcoding is an efficient, credible, and powerful approach to biomonitoring.