"The purpose of this paper is to present a sketch of the life of and a study of the novels written by a man who has used the theme of medicine, in combination with history and religion, to achieve considerable success. Since his first book came off the press in 1941, Frank G. Slaughter has had published eighteen novels that with only two exceptions dealt with medicine. Of the two exceptions, one was concerned with religion and the other with history, thereby maintaining the already assured formula for success"--Introduction. / Typescript. / "August, 1955." / Submitted to the Graduate Council of Florida State University in partial fulfillment of the requirements for the degree of Master of Science." / Advisor: Harold E. Moreland, Professor Directing Paper. / Includes bibliographical references (leaves 38-40).

Myxozoans are cnidarian parasites of primarily freshwater and marine fish, with some being important pathogens of aquacultured fish species worldwide. Their life cycles have waterborne actinospores released from aquatic annelid definitive hosts and myxospore stages in fish intermediate hosts. In the southeastern United States, catfish aquaculture is burdened by annual losses to a myriad of infectious diseases. Henneguya ictaluri, the causative agent of proliferative gill disease in channel catfish Ictalurus punctatus and female channel catfish x male blue catfish Ictalurus furcatus hybrids, is the most commonly diagnosed parasitic disease of catfish in Mississippi. Other myxozoans infect these ictalurid fish, but their impact on catfish production is unknown. Surveys of actinospores from the oligochaete Dero digitata and myxospore stages from fish revealed an unexpected diversity for these production systems. Six genetically distinct actinospores representing four collective groups were observed from D. digitata. Herein, two novel Henneguya spp. are described from the gills and a novel Unicauda sp. is described from the intestinal tract of channel catfish. One Henneguya sp. was linked to its actinospore stage and represents the fourth known life-cycle in the genus. In addition to catfish, smallmouth buffalo Ictiobus bubalus polycultured with catfish were examined and two Myxobolus spp. were characterized from the gills. Phylogenetic analyses strongly support a clade of ictalurid Henneguya spp. and a clade of catostomid Myxobolus spp. Although diverse, H. ictaluri is the only myxozoan in catfish attributed to significant losses. With no feasible method of control or treatment, investigations into less susceptible fish were initiated and showed promise. Infectivity trials characterizing H. ictaluri development in channel, blue, and hybrid catfish were performed. Channel catfish were suitable hosts with myxospores developing in the gills by six weeks and persisting for at least 14 weeks. In hybrid catfish arrested or limited development was observed with no pseudocysts observed during Trial 1 and only two at 14 weeks during Trial 2. These results may suggest a possible way of decreasing losses attributed to PGD through hybrid catfish monoculture or fish crop rotation to reduce the number of infectious myxospores released into the pond.

Proliferative gill disease (PGD) caused by the myxozoan parasite Henneguya ictaluri is one of the most devastating parasitic infections in channel catfish aquaculture. Currently, there is no effective treatment for H. ictaluri and the unpredictable outbreaks can result in 100% mortality. Management strategies have been developed to prevent losses in newly stocked fingerlings by evaluating the PGD status of a pond prior to stocking, which is difficult since resident fish may not show clinical signs even when actinospore levels are lethal to naive fish. Current diagnostic methods are limited to the identification of an active infection and methods of predicting potential outbreaks have several limitations. The PGD status of a pond to be stocked can be determined using sentinel fish exposures which are labor intensive and require a source of parasite free fish. These limitations necessitated the development of more rapid and efficient means of determining actinospore concentrations to determine the risk of losing fish prior to stocking. The development of a quantitative real-time polymerase chain reaction (QPCR) assay provided a more rapid, sensitive and quantitative method of diagnosing active infections and also provides a means to predict potential PD outbreaks and determine the PGD status of a pond prior to stocking. Another approach in the control of this parasite is the identification of a less susceptible culturable species or to identify traits that could be targeted in a selective breeding program. Challenge studies have shown that the closely related blue catfish (Ictalurus furcatus) does not exhibit as severe an inflammatory response to H. ictaluri and mortalities are significantly lower than in channel catfish. Comparisons of PGD severity and H. ictaluri infection in channel catfish, blue catfish and channel x blue catfish backcross hybrids by gross examination, histopathology and the newly developed H. ictaluri real-time PCR (QPCR) assay supported previous research suggesting the life cycle of the parasite can not be completed as efficiently through the blue catfish host. This dissertation describes the development and validation of a QPCR assay to detect H. ictaluri in both fish tissues and environmental samples and the application of this assay in both research and production settings.

<p> Serotonin positive (5HT+) and tyrosine hydroxylase positive (TH+) cells were identified using fluorescent immunocytochemical methods and quantified in the gill epithelium of six species of fish. 5HT+ cells were located in the filament epithelium in contact with the basal lamina on the efferent side, and in the lamellar epithelium where they were occasionally exposed to the external milieu. Thus, these cells appear to represent two populations of neuroepithelial cells (NEC) as proposed in other studies. In trout, bass and killi fish, NECs were revealed exclusively in the primary epithelium. In tilapia, NECs were located exclusively in the secondary epithelium, whereas in perch and zebrafish they occurred in both epithelial layers. The two types of NECs varied in number both within and among the species. Seasonal comparisons of NECs in perch revealed a decrease in cell density in the filament between July and November, though there was no significant difference in the density of NECs in the lamellae. TH+ cells were identified in perch, zebrafish and killi fish. In zebrafish TH+ cells occurred in similar numbers to 5HT+ cells, and were generally present in similar locations. It is likely that in this case, many of the labelled cells were positive for both markers. In two of the species, perch and killi fish, the density and distribution was such that the TH+ cells and 5HT+ cells were unlikely to be the same. A quantitative comparison of total catecholamine (CA) stores, using high performance liquid chromatography (HPLC), revealed that gill tissues in general contained higher levels of epinephrine (EPI) than norepinephrine (NOR) and dopamine (DOP). Finally, attempts were made to determine whether NECs would survive in 2-4 day old cultures of dispersed gill cells from perch, using immunocytochemical labelling for 5HT. A few successful cases are presented.</p> / Thesis / Master of Science (MSc)

Goldfish undergo an adaptive morphological change in their gills involving the reversible growth and loss of a mass of cells (interlamellar cell mass, ILCM) in between the lamellae depending on oxygen demand, which can be altered by the environment or metabolic demands of the individual. The ILCM contributes to decreased passive Na+ efflux across the gill. Active uptake is maintained by the re-distribution of the ionocytes expressing Na+-uptake relevant genes (NHEs and H+-ATPase) to the outer edge of the ILCM where they can establish contact with the external environment and/or lamellar epithelium. This adaptation is thought to be partly responsible for the extreme anoxia tolerance demonstrated by goldfish, which they experience on a seasonal basis living in a pond environment. Hypoxia and hypercapnia are frequently encountered in such freshwater environments and as such, the effect of the ILCM on the capacity for acid-base regulation was evaluated. Differences in the time course of acid excretion to the environment without effect on systemic pH regulation were likely the result of the ILCM.

Goldfish undergo an adaptive morphological change in their gills involving the reversible growth and loss of a mass of cells (interlamellar cell mass, ILCM) in between the lamellae depending on oxygen demand, which can be altered by the environment or metabolic demands of the individual. The ILCM contributes to decreased passive Na+ efflux across the gill. Active uptake is maintained by the re-distribution of the ionocytes expressing Na+-uptake relevant genes (NHEs and H+-ATPase) to the outer edge of the ILCM where they can establish contact with the external environment and/or lamellar epithelium. This adaptation is thought to be partly responsible for the extreme anoxia tolerance demonstrated by goldfish, which they experience on a seasonal basis living in a pond environment. Hypoxia and hypercapnia are frequently encountered in such freshwater environments and as such, the effect of the ILCM on the capacity for acid-base regulation was evaluated. Differences in the time course of acid excretion to the environment without effect on systemic pH regulation were likely the result of the ILCM.

Goldfish undergo an adaptive morphological change in their gills involving the reversible growth and loss of a mass of cells (interlamellar cell mass, ILCM) in between the lamellae depending on oxygen demand, which can be altered by the environment or metabolic demands of the individual. The ILCM contributes to decreased passive Na+ efflux across the gill. Active uptake is maintained by the re-distribution of the ionocytes expressing Na+-uptake relevant genes (NHEs and H+-ATPase) to the outer edge of the ILCM where they can establish contact with the external environment and/or lamellar epithelium. This adaptation is thought to be partly responsible for the extreme anoxia tolerance demonstrated by goldfish, which they experience on a seasonal basis living in a pond environment. Hypoxia and hypercapnia are frequently encountered in such freshwater environments and as such, the effect of the ILCM on the capacity for acid-base regulation was evaluated. Differences in the time course of acid excretion to the environment without effect on systemic pH regulation were likely the result of the ILCM.

Goldfish undergo an adaptive morphological change in their gills involving the reversible growth and loss of a mass of cells (interlamellar cell mass, ILCM) in between the lamellae depending on oxygen demand, which can be altered by the environment or metabolic demands of the individual. The ILCM contributes to decreased passive Na+ efflux across the gill. Active uptake is maintained by the re-distribution of the ionocytes expressing Na+-uptake relevant genes (NHEs and H+-ATPase) to the outer edge of the ILCM where they can establish contact with the external environment and/or lamellar epithelium. This adaptation is thought to be partly responsible for the extreme anoxia tolerance demonstrated by goldfish, which they experience on a seasonal basis living in a pond environment. Hypoxia and hypercapnia are frequently encountered in such freshwater environments and as such, the effect of the ILCM on the capacity for acid-base regulation was evaluated. Differences in the time course of acid excretion to the environment without effect on systemic pH regulation were likely the result of the ILCM.

The occurrence of ichthyofauna on the territory of the Jizera Mountains (Northern Bohemia, Czech Republic) is limited, besides other factors, by the water chemistry. pH value and pH-related concentration of inorganic monomeric aluminum (Ali) are crucial parameters of the water, determining the survival of fish in streams. Low pH and high concentrations of Ali cause severe damages to fish gills. The thesis deals with the influence of the water chemistry on distribution of ichthyofauna in the Jizera Mts and gill morphology in brook charr (Salvelinus fontinalis), a dominant fish species of the upper plateau of the mountains. The results show that the species diversity of fish in acidified streams is lower than that in streams without meaningful acidic episodes. Histopathological analyses of gills of brook charr individuals from an acidified stream confirm, that low pH values and high concentrations of Ali have a cummulative effect and lead to degenerative changes on gills. Key words: The Jizera Mountains, ichthyofauna, brook charr, gill morphology, water chemistry, toxic aluminium, acidification

O Ucides cordatus é um caranguejo que habita mangues. Estas regioes podem ser contaminadas com o cádmio, através de dejetos de indústrias e descarte de pilhas e baterias, atingindo o animal por meio de suas brânquias. Assim, o objetivo da dissertação foi verificar a cinética de transporte do cádmio com o uso de inibidores específicos, e sua interação com o cálcio, zinco e ATP. Para isso, as brânquias foram separadas em anteriores e posteriores, e as células foram dissociadas pelo método de dissociação enzimática; a seguir, as células foram marcadas com Fluo 3 am e realizada a cinética de transporte no espectrofluorímetro, adicionando-se concentrações de 0,5, 1,0 e 1,5&mu;M de sulfato de cádmio (CdSO4), cloreto de cádmio (CdCl2), cádmio + cálcio, cádmio + zinco, cálcio e zinco para verificar a interação do Cd com cálcio ou zinco. Posteriormente, houve a adição dos inibidores como vanadato, amilorida, ouabaina e BAPTA às células, e, novamente, efetuou-se o transporte de cádmio nas concentrações 0,5, 1,0 e 1,5&mu;M. Em seguida, adicionou-se ATP e efetuou-se o transporte de cádmio nas mesmas concentrações citadas anteriormente. Os resultados mostraram um transporte de cádmio sigmoidal, quando em presença de CdSO4, e um pico de 5 segundos de entrada de cádmio e uma posterior queda, quando em presença de CdCl2; além de uma provável competição entre o cádmio e o cálcio e entre o cádmio e o zinco. Observou-se também uma efetiva ação dos inibidores, indicando que o cádmio possivelmente entrou na célula através de canais de cálcio, trocadores Na+/H+ e trocadores aniônicos dependentes ou não de Na+. Percebeu-se ainda que o ATP não influenciou na entrada de cádmio, ou seja, aparentemente não existe uma Cd ATPase nas células branquiais de Ucides cordatus / Ucides cordatus is a crab that lives in mangroves. These regions may be contaminated with cadmium by industrial wastes and disposal of batteries, reaching the animal through its gills, being a toxic metal. The aim of the study was to investigate the kinetics of cadmium transport using specific inhibitors, and its interaction with calcium, zinc and ATP. For this, the gills were separated in anterior and posterior regions, and their cells were dissociated by enzymatic dissociation. After, the cells were labeled with Fluo 3 am and the kinetics of transport of cadmium were performed in a spectrofluorimeter by adding CdSO4 and CdCl2 at concentrations of 0.5, 1.0 and 1.5&mu;M. For the interaction experiments Cd was added as, cadmium + calcium, cadmium + zinc, calcium and zinc alone. A subsequent experiment, addition of inhibitors such as vanadate, amiloride, ouabain and BAPTA were performed, and again, the transport of cadmium was studied at concentrations of 0.5, 1.0 and 1.5 &mu;M. Then, ATP was added and the transport of cadmium was performed in the same concentrations mentioned above. Results showed a sigmoidal curve, when in presence of CdSO4, and a peak of 5 seconds for cadmium uptake and a posterior decline when in the presence of CdCl2. In addition, probably occurred a competition between calcium and cadmium and between cadmium and zinc, and here was also an effective action of some inhibitors, indicating that cadmium probably entered the cell by calcium channels, Na+/H+ exchangers and exchangers anionic inorganic dependent of Na+ or not. ATP had no effect for cadmium influx, suggesting that there is no Cd ATPase present in gill cells of Ucides cordatus