Since being first reported in the late 1980ies, the Enterobacterium Edwardsiella ictaluri has rose in prevalence to become one of the two most damaging pathogens affecting the channel catfish industry. Despite this significance of the pathogen, understanding of the development of the disease, especially its route of entry into the host and the earlier stages of the infection, is still incomplete. A series of challenges were conducted using bioluminescent E. ictaluri either by infecting fish through immersion or topical application of the bacteria directly on the intact or abraded epithelium. This showed that abraded fish developed septicemia and died faster than non-abraded ones. Furthermore, results from a co-habitation challenge suggested that the bacterium induced septicemia through the skin instead of becoming water-borne. Finally, a histological technique was developed allowing the determination that the bacteria radiated from the initial skin infection site and penetrated deeper into the tissue as the challenge progressed. These results all suggest that site of abrasion on the skin can act as a route of entrance for the pathogen into the fish, a fact never previously reported. Transposon mutagenesis was also performed to construct a library of 1728 mutants. Screening of this library allowed us to identify 16 genes which inactivation lead to a decrease in the bacterium ability to colonize the epithelium or cause mortality. Sequencing of these genes allowed the identification of RstA/B, a regulator of invasion genes in Salmonella enterica Typhimurium, a putative ribonuclease, similar to a Shigella protein regulating the expression of adhesin and a protein that constitutes the second member of a newly discovered adhesin family. Finally, to investigate the development of the infection, fish were infected by bioluminescent E. ictaluri and sampled at various time points. At each time point, nine organs (gills, muscles, intestine, spleen, liver, stomach, heart, head kidney and trunk kidney) were sampled, and their bioluminescence was measured and half of these organs were homogenized, serial diluted, and plate counts determined. This allowed confirmation of a complex disease pathogenesis during ESC involving a period of intense reproduction in the spleen, anterior and posterior kidneys followed by a sharp increase in the levels of bacteria in the blood.

Over the last 3 decades, freshwater aquaculture has become one of the most important food industries. However the constant introduction of a reduced number of very successful species for aquaculture has been identified as one of the main activities related to the alarming decline of fish biodiversity worldwide. This issue has raised awareness amongst the scientific community, governmental authorities and the general public towards freshwater fish biodiversity. This new awareness has promoted the development of “green” markets and environmentally friendly strategies, aiming for a reliable production of protein sources. The development of native species aquaculture has been presented as a strong alternative for sustainable aquaculture and the protection of biodiversity. However, it seems clear that unplanned native species aquaculture developments can be as detrimental on local biodiversity as the introduction of exotic fish, if not more dangerous. Therefore, the advantages and disadvantages of native species aquaculture have to be clearly analysed before any aquaculture development. This study aimed to establish a philosophical background regarding the use of native fish species in aquaculture in contrast to the introduction of exotic species that may compete for a similar niche as food in local markets. The main ecological impacts that exotic fish species may have on natives, such as competition, predation, and hybridization were discussed. In addition, a well planned native species Aquaculture Strategy for the Protection of Biodiversity was produced, at catchment level, within a Geographic Information System (GIS). For the development of the native species aquaculture strategy in central Mexico, four species of Atherinids (Chirostoma estor, C. Jordani, C. promelas and Atherinella balsana) and two species of native Ictalurids (Ictalurus balsanus and Ictalurus dugesii) were included in this study. These six species are relatively new to aquaculture and they were selected on the basis of their importance in local fisheries and markets in their native basins of the Lerma-Santiago and Balsas rivers. Both of these basins are of great importance in central Mexico, not only because of their biodiversity but also because of their high human population densities and socio-economic status. The use of Geographic Information Systems was a fundamental factor in the development of the native species aquaculture strategy at catchment level, consisting of site suitability models (SSM) for each species in their corresponding native catchments. Overall, SSM identified 13,916 km2 and 11,178 km2 highly suitable for aquaculture of the studied Atherinids and Ictalurids respectively, based on Water, Soil and Terrain, Infrastructure and Risk sub-models. A set of predictive species distribution models (PSDM), which related ecological characteristics for each studied species with relevant environmental and topographic parameters into a GIS, were also produced. Such models were developed for the establishment of potential natural ranges of distribution for each species, as well as their potential to become exotic in new environments, as a potential for invasion model (PI). Based on a partial verification, both PSDM and PI models produced results that were satisfactorily consistent with the known distribution of each modelled species. The combination of SSM and PSDM produced an Aquaculture Strategy for the Protection of Biodiversity model (ASPB) which identified the most environmentally friendly suitable areas for aquaculture sites. In contrast, the combination of the SSM with PI models into an ASPB model identified the site suitability potential for non-native species that are genetically close to native ones, in an attempt to reduce the known impacts that exotic species have on local biodiversity. In this way the ASPB model identified 7,651 km2 suitable for aquaculture of I. balsanus in its native Balsas basin and 15,633 km¬2 suitable for aquaculture of the non-native I. dugesii. ASPB models were produced for all the studied species. The final results were used to produce a set of guidelines for the development of sustainable aquaculture of native species at catchment level that cover genetic and ecological implications, as well as a well planned decision making tool produced in a GIS.

Master of Science / Division of Biology / Martha E. Mather / Understanding how and why fish distribution is related to specific habitat characteristics underlies many ecological patterns and is crucial for effective research and management. Blue Catfish, Ictalurus furcatus, are an important concern for many fisheries agencies; however, lack of information about their distribution and habitat use remains a hindrance to proper management. Here, over all time periods and across months, I quantified Blue Catfish distribution and environmental correlates of distribution in Milford Reservoir, the largest reservoir in Kansas. I tested relationships among acoustically tagged Blue Catfish and three groups of variables postulated to influence Blue Catfish distribution in the literature (i. localized microhabitat variables, ii. larger-scale mesohabitat variables, iii. biotic variables). Blue Catfish were consistently aggregated in two locations of the reservoir across five months during summer and fall, 2013. Using multiple linear regression and an information theoretic model selection approach, consistent correlates of distribution included localized, microhabitat variables (i.e., dissolved oxygen, slope) larger-scale, mesohabitat variables (i.e., distance to channel, river kilometer from the dam) and a biotic variable (i.e., Secchi depth). This research identified which 5 of the 12 variables identified in the literature were most influential in determining Blue Catfish distribution. As a guide for future hypothesis generation and research, I propose that Blue Catfish distribution was driven by three ecologically-relevant tiers of influence. First, Blue Catfish avoided extremely low dissolved oxygen concentrations that cause physiological stress. Second, Blue Catfish aggregated near the channel, an area of bathymetric heterogeneity that may offer a foraging advantage. Third, Blue Catfish aggregated near low Secchi depths, shown here to be associated with increased productivity and prey abundance. Building on my results, future research into the distribution and habitat use of Blue Catfish should incorporate aggregated distributions of fish into research designs, focus on how both small and large scale relationships interact to produce patterns of distribution, and explore further the mechanisms, consequences, and interactions among the three tiers of influence identified here.

Acoustically, teleost swimbladders have been considered resonant underwater bubbles. Contrary results indicating that bladders are tuned less sharply than such a bubble have been explained by damping of surrounding fish tissue. Recent findings in toadfish and weakfish, however, suggest that the bladder is a highly damped structure and that the frequency of the fish sounds is deternlined as a forced response to sonic muscle movement rather than by resonance of the bladder. In this study I examined acoustics and material properties of swimbladders in Tilapia (an auditory generalist) and the channel catfish (an auditory specialist). The swimbladder was struck with a piezoelectric impact hammer. Amplitude and timing characteristics of bladder sound and displacement were compared for strikes of different amplitudes. Most of the first cycle of sound occurs during swimbladder compression, indicating that the bladder rapidly contracts and expands as force increases during the strike. Harder hits are shorter in duration generate a similar displacement duration with an increasing number of shorter cycles and a 12-15 dB increase in sound amplitude. The frequency spectrum is broad, and the dominant frequency is driven by the strike and not the natural frequency of the bladder. The displacement waveform varies between species catfish exhibit a greater structural stiffness and lower amplitude movement and higher sound amplitude for an equivalent hammer strike. Material properties (peak load, stress, strain, Young's modulus), fiber direction (catfish only), and structural stiffiless of bladders exhibit various patterns suggesting that the bladder walls are not uniform structures. Additionally thickness varies regionally. Notably Young's modulus in both species is similar despite large difference in stress and strain. The modulus of catfish bladder increased 1600-fold when dried. Finally the bladder of both species had a high water content averaging about 70%. These data suggest that viscous damping caused by water in the bladder wall is a major factor responsible for acoustic properties of the teleost swimbladder.

Thesis (Ph.D.)--Mississippi State University. Department of Agricultural and Biological Engineering. / Title from title screen. Includes bibliographical references.

The commercial production of channel catfish (Ictalurus punctatus) is major industry in Mississippi. Infections of channel catfish with the digenetic trematode Bolbophorus damnificus have often been associated with heavy economic losses in the industry. To efficiently control transmission of this trematode, the avian hosts need to be identified. In the first study, two American white pelicans, two double-crested cormorants, two great blue herons, and two great egrets were fed channel catfish infected with B. damnificus metacercariae. The presence of Bolbophorus damnificus ova in pelican feces at three days post infection (dpi) indicated the pelicans had patent infections. Mature B. damnificus were recovered from the intestines of both pelicans at 21 dpi. No B. damnificus infections were observed in the other bird species. In a second study, 33 American white pelicans, 34 double-crested cormorants, 35 great blue herons, and 32 great egrets were collected in the Mississippi Delta. The prevalence of B. damnificus in the American white pelican was 93.9%, with an average of 158 B. damnificus found per bird (range 0-681). Bolbophorus damnificus was not found in any of the other bird species. The results of these two studies confirm that the AWPE is the only proven natural host for B. damnificus. In a third study, two previously undescribed cercariae were found infecting rams-horn snails in commercial catfish ponds. In challenge studies, channel catfish were exposed to both cercariae types. Only one type of cercariae (type I) was infective to channel catfish. The first evidence of type I metacercariae was seen histologically at 14 dpi and grossly at 21 dpi. Development continued until 120 dpi, when both gross examination and histology suggested that the metacercariae were mature. The type I metacercariae appeared to cause little host damage. Molecular analysis of the 18S rRNA gene region indicated that the type I cercariae and metacercariae may be a species of Clinostomum. The data generated in these three studies provides additional information that can be used in the development of efficacious management schemes to control digenetic trematodes infecting commercial catfish.

The catfish industry suffers losses primarily due to enteric septicemia of catfish and columnaris disease caused by Edwardsiella ictaluri and Flavobacterium columnare, respectively. Understanding the host-pathogen interactions is vital for prevention and eradication of these diseases. Hence, the overall objective of this study was to analyze whole cell proteomes of these two bacteria, and to determine the changes in E. ictaluri protein expression against in vitro iron-restriction and host serum treatment. High-throughput proteomic analysis of these bacteria was conducted using two-dimensional liquid chromatography followed by electrospray ionization tandem mass spectrometry (2-D LC ESI MS/MS) and two-dimentional gel electrophoresis coupled with matrix-assisted laser desorption/ionization time-oflight mass spectrometry (2-DE MALDI TOF/TOF). Identified proteins were clustered into functional groups using clusters of orthologous groups, and subcellular locations as well as possible functional relationships were determined. A total of 788 unique E. ictaluri and 621 unique F. columnare proteins were identified, which represented 12 and 28 pathways, respectively. Vertebrate hosts tend to chelate free iron of their body and make the environment hostile for bacteria. Hence, reduced availability of iron may cause significant stress for pathogens and is considered a signal that leads to alteration in virulent gene expression. Similarly, E. ictaluri might use the catfish blood stream effectively for quick systemic invasion. Hence, exposure to catfish serum components might reveal the ability of E. ictaluri to protect against host defense mechanisms. Using two-dimensional difference gel electrophoresis, responses of E. ictaluri due to in vitro iron-restriction and host serum treatment were determined. A total of 50 and 19 proteins were identified to be differentially expressed due to in vitro iron-restriction and catfish serum treatment, respectively. Among the differentially expressed proteins, several putative virulent determinants, immunogenic proteins, chaperones, and housekeeping genes were noted. To initiate functional studies, four differentially expressed E. ictaluri genes (lamB, glyS, malE, and sdhA) were mutated by inrame deletion. Results from this study provided experimental evidence for many predicted proteins. In addition, identification of differentially expressed proteins provided targets for further functional analysis, which could help elucidate pathogenic mechanisms of E. ictaluri.