Many Gopher Tortoise populations are in steep decline throughout Florida, and various measures have been attempted to curb the trend. One such measure is to relocate tortoises to protected recipient sites on private lands. The majority of private lands in Florida are used for cattle, however, production and the effect of cattle production on tortoises is not known. Here, I tested six parameters of tortoise behavior by monitoring 1403 gopher tortoises released at the Barthle Brothers Ranch, Pasco County Florida, between August 2009 and December 2012. The parameters tested were (1) burrow density, (2) burrow spacing, (3) burrow relocation, (4) body condition, (5) individual growth rate, and (6) recruitment (addition of young to the population). I used telemetry techniques to observe movement and burrow placement as it related to cattle activity and burrow impacts, and collected morphological data to determine changes in body condition and growth. I used burrow surveys and analyzed movement patterns to interpret the propensity for tortoises to place burrows where cattle may or may not congregate. Lastly, I investigated recruitment of juveniles into the population and followed the mortality of resident and relocated tortoises in all treatment plots. I found that burrow density, distance moved when relocating to new burrows, and avoidance of cattle were not distinguishably different within or between the plots. Burrow relocation, however, was more frequent outside the exclosures. The change in body condition did not differ between males and female or resident and relocated individuals. Females within the exclosure did not grow at a rate different than those outside the exclosure although translocated females grew faster than resident females. Densities of non-adult burrows inside the exclosures were not different then densities outside the exclosure. Eleven percent of tortoises relocated to the ranch died during the project. Although we have no evidence that cattle and tortoise cannot successfully coexist, a number of circumstances prevented rigorous testing of our hypotheses, predominately the failure of the silt fence used to enclose the treatment plots. Using a trespass-proof perimeter fence would allow a better assessment of the actual interaction between the cattle and tortoises and may shed new light on the lack of recruitment and the decline of juvenile tortoises relocated to the ranch. Without recruitment of individuals back into a population, or the persistent of reproducing adults within the population, any efforts to curb the downward trend in gopher tortoise numbers by relocating tortoises to actively grazed pasture is futile.

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Abstract not available

All of New Zealand's five endemic species of mudfish (Neochanna) are threatened, and translocation has been recommended as an option for conservation. This research undertakes a translocation of black mudfish (Neochanna diversus) into wetland margins of Lake Kaituna, in the Waikato region, and addresses research questions applicable to improving translocation success. Results from this research are intended to aid possible future translocations of the more threatened Northland mudfish (Neochanna heleios) and other genetically distinct populations of black mudfish. Captive rearing of juveniles collected from the wild is currently the most feasible option for sourcing translocation stock. Mudfish juveniles (greater than 25 mm T.L.) had greater survival rates, compared to mudfish fry less than 25 mm T.L. Mudfish growth was far greater when fed on a combined diet of brine shrimp (Artemia salina) and white worms (Enchytraeus albidus) than when fed exclusively on brine shrimp. Temperature was found to have a small effect on mudfish growth, with a slightly greater growth in fish at 15 C than those at 10 C. The introduced species Gambusia affinis has been the subject of concern for mudfish conservation and commonly found at wetland sites suitable for mudfish translocation. Found to prey on mudfish fry and eggs in aquaria, it was important to determine the effects of Gambusia density prior to undertaking a translocation to a location where Gambusia were present. Investigations were made into the effect of Gambusia density on black mudfish juveniles in 9 outdoor mesocosms. Increasing Gambusia density was found to have an inhibitory effect on black mudfish growth. This may be due to increased competition for food, a theory supported by analysis of zooplankton communities, where, in the presence of Gambusia, large zooplankton had been removed and smaller rotifers flourished. Monitoring programmes are required to assess any impacts or improvements of mudfish populations, including those created by translocation. A Gee minnow trapping programme in outdoor mesocosms was conducted to test the reliability of traps, finding that water depth, mudfish density, mudfish memory and trap shyness had no effect on the trapability of mudfish. Trap position was found to have the most significant effect, with a greater number of mudfish caught when traps were set overnight at the surface than when set on the bottom of mesocosms. Black mudfish adults and juveniles were translocated into 18 pools (~1 m diameter) on the wetland margins of Lake Kaituna in September 2006, followed by monthly monitoring. Water quality monitoring and an assessment of hydrology and vegetation was undertaken. Habitat characterisation was found to be a key factor, with correlations between water quality data and trapping results finding fewer fish remaining in pools with less suitable characteristics for mudfish (e.g. high turbidity and conductivity). Other species were found to have a large impact, with predation by shortfinned eels (Anguilla australis) thought to have eliminated mudfish from some pools. In addition fewer mudfish were caught in pools with Gambusia, possibly due to increased competition.

La toxine diphtérique (DT) une toxine bactérienne sécrétée par Corynebacterium diphtheriae. Lors de l'intoxication d'une cellule, le domaine de translocation (T) de la DT s'insère dans la membrane à pH acide et assiste la translocation du domaine catalytique (C) dans le cytosol. Le domaine T adopte une conformation en molten globule (MG) et devient compétent pour l'interaction membranaire. Nous avons identifié par mutagenèse les résidus du domaine T dont la protonation favorise la formation de l'état MG en solution et l'interaction membranaire. Les résultats montrent que la protonation concertée des six histidines du domaine T est impliquée dans la formation de l'état MG en solution. La paire His223-257 et l'His251 ont un effet prépondérant dans la formation de l'état molten globule en solution, alors que la paire His322-323 (mais également His251) est davantage impliquée dans l'interaction avec la membrane, et en particulier la liaison à la membrane.<br />Nous avons étudié les changements de conformation des deux domaines C et T dans une protéine CT correspondant à la DT tronquée de son domaine R, afin de comprendre les effets du lien covalent sur les conformations respectives de C et T en fonction du pH. Les mutants CTW50/153F et CTW206/281F ont permis de suivre la fluorescence des Trp de chaque domaine dans la protéine CT. Les<br />résultats montrent que le domaine T dirige la translocation de C dans les premières étapes de la translocation (formation de l'état MG, liaison et insertion membranaire), et qu'il pourrait jouer un rôle de chaperon en stabilisant l'état MG du domaine C.

Il existe différentes techniques de manipulation et de détection de molécules uniques. Parmi elles, la mesure du courant ionique traversant un pore nanométrique unique est la plus ancienne. Le travail effectué dans cette thèse utilise cette technique et s'articule autour de deux objectifs. Tout d'abord, l'utilisation d'un nanopore unique obtenu par attaque de trace permet d'imiter les expériences déjà faites dans la littérature avec d'autres types de pores. Certains phénomènes, tels que la rectification par exemple, ont pu être observés, mais d'autres comme la translocation n'ont pu l'être. Ensuite, une étude du bruit de conductance électrique démontre l'existence d'un phénomène qui n'avait jusqu'alors pas été soulevé dans la littérature. En effet, ce bruit ne peut pas être imputé aux fluctuations des caractéristiques de la géométrie du pore ou de sa paroi, mais plutôt à des effets coopératifs sur la mobilité des ions dans un milieu confiné.

<p>Metal contaminated stormwater, i.e. surface runoff in urban areas, can be treated in percolation systems, ponds, or wetlands to prevent the release of metals into receiving waters. Plants in such systems can, for example, attenuate water flow, bind sediment, and directly accumulate metals. By these actions plants affect metal mobility. This study aimed to examine the accumulation of Zn, Cu, Cd, and Pb in roots and shoots of plant species common in stormwater areas. Furthermore, submersed plants were used to examine the fate of metals: uptake, translocation, and leakage. Factors known to influence metal accumulation, such as metal ion competition, water salinity, and temperature, were also examined. The following plant species were collected in the field: terrestrial plants – <i>Impatiens parviflora</i>, <i>Filipendula ulmaria</i>, and <i>Urtica dioica</i>; emergent plants –<i> Alisma-plantago aquatica</i>, <i>Juncus effusus</i>, <i>Lythrum salicaria</i>, <i>Sagittaria sagittifolia</i>, and <i>Phalaris arundinacea</i>; free-floating plants – <i>Lemna gibba</i> and <i>Lemna minor</i>; and submersed plants – <i>Elodea canadensis</i> and <i>Potamogeton natans</i>. Furthermore, the two submersed plants, <i>E. canadensis</i> and <i>P. natans</i>, were used in climate chamber experiments to study the fate of the metals in the plant–water system.</p><p>Emergent and terrestrial plant species accumulated high concentrations of metals in their roots under natural conditions but much less so in their shoots, and the accumulation increased further with increased external concentration. The submersed and free-floating species accumulated high levels of metals in both their roots and shoots. Metals accumulated in the shoots of <i>E. canadensis</i> and <i>P. natans</i> derived mostly from direct metal uptake from the water column.</p><p>The accumulation of Zn, Cu, Cd, and Pb in submersed species was in general high, the highest concentrations being measured in the roots, followed by the leaves and stems, <i>E. canadensis</i> having higher accumulation capacity than <i>P. natans</i>. In <i>E. canadensis</i> the Cd uptake was passive, and the accumulation in dead plants exceeded the of living with time. The capacity to quickly accumulate Cd in the apoplast decreased with successive treatments. Some of the Cd accumulated was readily available for leakage. In <i>P. natans,</i> the presence of mixtures of metal ions, common in stormwater, did not alter the accumulation of the individual metals compared to when presented separately. It is therefore, proposed that the site of uptake is specific for each metal ion. In addition cell wall-bound fraction increased with increasing external concentration. Further, decreasing the temperature from 20ºC to 5ºC and increasing the salinity from 0‰ to 5‰ S reduced Zn and Cd uptake by a factor of two.</p><p>In <i>P. natans</i> the metals were not translocated within the plant, while in<i> E. canadensis </i>Cd moved between roots and shoots. Thus,<i> E. canadensis</i> as opposed to <i>P. natans</i> may increase the dispersion of metals from sediment via acropetal translocation. The low basipetal translocation implies that neither <i>E. canadensis</i> nor <i>P. natans</i> will directly mediate the immobilisation of metal to the sediment via translocation.</p><p>To conclude, emergent and terrestrial plant species seem to enhance metal stabilization in the soil/sediment. The submersed plants, when present, slightly increase the retention of metals via shoot accumulation.</p>

Metal contaminated stormwater, i.e. surface runoff in urban areas, can be treated in percolation systems, ponds, or wetlands to prevent the release of metals into receiving waters. Plants in such systems can, for example, attenuate water flow, bind sediment, and directly accumulate metals. By these actions plants affect metal mobility. This study aimed to examine the accumulation of Zn, Cu, Cd, and Pb in roots and shoots of plant species common in stormwater areas. Furthermore, submersed plants were used to examine the fate of metals: uptake, translocation, and leakage. Factors known to influence metal accumulation, such as metal ion competition, water salinity, and temperature, were also examined. The following plant species were collected in the field: terrestrial plants – Impatiens parviflora, Filipendula ulmaria, and Urtica dioica; emergent plants – Alisma-plantago aquatica, Juncus effusus, Lythrum salicaria, Sagittaria sagittifolia, and Phalaris arundinacea; free-floating plants – Lemna gibba and Lemna minor; and submersed plants – Elodea canadensis and Potamogeton natans. Furthermore, the two submersed plants, E. canadensis and P. natans, were used in climate chamber experiments to study the fate of the metals in the plant–water system. Emergent and terrestrial plant species accumulated high concentrations of metals in their roots under natural conditions but much less so in their shoots, and the accumulation increased further with increased external concentration. The submersed and free-floating species accumulated high levels of metals in both their roots and shoots. Metals accumulated in the shoots of E. canadensis and P. natans derived mostly from direct metal uptake from the water column. The accumulation of Zn, Cu, Cd, and Pb in submersed species was in general high, the highest concentrations being measured in the roots, followed by the leaves and stems, E. canadensis having higher accumulation capacity than P. natans. In E. canadensis the Cd uptake was passive, and the accumulation in dead plants exceeded the of living with time. The capacity to quickly accumulate Cd in the apoplast decreased with successive treatments. Some of the Cd accumulated was readily available for leakage. In P. natans, the presence of mixtures of metal ions, common in stormwater, did not alter the accumulation of the individual metals compared to when presented separately. It is therefore, proposed that the site of uptake is specific for each metal ion. In addition cell wall-bound fraction increased with increasing external concentration. Further, decreasing the temperature from 20ºC to 5ºC and increasing the salinity from 0‰ to 5‰ S reduced Zn and Cd uptake by a factor of two. In P. natans the metals were not translocated within the plant, while in E. canadensis Cd moved between roots and shoots. Thus, E. canadensis as opposed to P. natans may increase the dispersion of metals from sediment via acropetal translocation. The low basipetal translocation implies that neither E. canadensis nor P. natans will directly mediate the immobilisation of metal to the sediment via translocation. To conclude, emergent and terrestrial plant species seem to enhance metal stabilization in the soil/sediment. The submersed plants, when present, slightly increase the retention of metals via shoot accumulation.

Bacteria that infect humans and animals face a hard combat with the host´s immune system and in order to establish infection, pathogenic bacteria has evolved mechanisms to avoid being cleared from the host tissue. Many Gram-negatives carry a Type 3 secretion (T3S) system that is used to deliver effector proteins (toxins) into host cells. The toxins exhibit a broad range of intra cellular effects that has in common that they increase the chances of the bacteria to establish infection, multiply in infected tissue or spread to other tissues or hosts. The object of this study was to analyse the mechanisms behind the T3S effectors delivery into target cells. Two bacterial proteins, LcrV and YopD, which are involved in the translocation of effectors were analyzed by mutagenesis. LcrV was found to affect the efficiency of the translocation, probably by determining the size of the pore in the target cell membrane through which the effectors pass. Truncated variants of the multi-functional YopD revealed that defined regions of the protein were important for pore-formation and translocation. Effectors and translocators were demonstrated to form complexes with free acyl chains (lipids) at the bacterial surface. These complexes –termed Yop-lipid complexes, (YLC)– are released from the surface and can act as discrete units that are able to promote translocation of effectors even when separated from the bacterium from which they originate. These findings shed new light on the process of effector translocation by Yersinia and possibly by other gram-negative bacterial pathogens with a similar T3S setup.

The Gram-negative bacterium Yersinia pseudotuberculosis employs a type 3 secretion system (T3SS) to establish infections. The T3SS translocates a diverse set of effector proteins directly into the host cells. The coordinate action of the translocated effectors blocks the innate immune system of the host and ensures extracellular proliferation of the bacterium. YopE is an essential effector that disrupts the actin cytoskeleton of infected host cells. This cytotoxicity is caused by the inactivation of RhoGTPases by the GTPase Activating Protein (GAP) activity of YopE. YopE was demonstrated to inactivate the RhoGTPases Rac1 and RhoA in vivo. However, Rac1 and RhoA inactivation was not a prerequisite for cytotoxicity or virulence. Thus, YopE must have additional targets during infection. Surprisingly, avirulent yopE mutants had lost the control of Yop expression in the presence of target cells and they all overtranslocated effectors. It appeared as if translocated YopE was able to control Yop expression and effector translocation via a feedback inhibition mechanism. This feedback inhibition was dependent on functional GAP activity. Translocation control could also be mediated by exogenous GAP activity, suggesting that effector translocation control might be a general property of all bacterial GAP proteins. Besides YopE, the regulatory protein YopK was also found to be involved in the effector translocation control process. Clearly, as demonstrated in virulence, the roles for YopE and YopK are intimately related.                       Further, YopE possesses a membrane localization domain (MLD) required for proper localization. A yopE∆MLD mutant had lost the feedback inhibition of YopE expression and was avirulent. Hence, the effector translocation control of YopE requires both proper localization as well as functional GAP activity.                                           In addition, fish keratocytes were established as a novel model system for Y. pseudotuberculosis infections. YopE was found to be the sole effector responsible for cytotoxicity towards the keratocytes. Further, induction of cytotoxicity required fully native YopE protein which indicated that the keratocytes would be useful as a sensitive model system for further studies of YopE mediated phenotypes. In summary, this thesis work has sought to unravel the multiple functions of translocated YopE. A novel role was elucidated where Yersinia utilizes translocated YopE to control the process of effector translocation into host cells. This regulatory control was connected to virulence in the mouse model of disease. Thus, perhaps YopE should be considered also as a regulatory protein besides being a classical effector.