Fungi for Future : Biotechnological application of anaerobic fungi

Der Verwendung Lignocellulose-haltiger Rohstoffe wird eine essentielle Rolle beim Übergang von einer auf fossilen Rohstoffen basierenden zu einer biobasierten Ökonomie beigemessen. Die für die biotechnologische und chemische Nutzbarmachung von Lignocellulose als Ausgangssubstrat nötige Vorbehandlung ist jedoch energieintensiv und/oder produziert toxische Abfälle und Nebenprodukte. Wasserstoff ist ein vielversprechender Energieträger für eine zukünftige Bioökonomie, der jedoch zurzeit hauptsächlich aus fossilen Rohstoffen gewonnen wird. Eine Alternative bietet die biologische Wasserstoffproduktion auf Basis von Lignocellulose, die jedoch zurzeit durch den hohen Energiebedarf der Lignocellulose-Vorbehandlung sowie niedriger Ausbeuten bisher eher unbedeutend ist. Anaerobe Pilze des Phylums Neocallimastigomycota können durch die Sekretion einer Vielzahl von Lignocellulose abbauenden Enzymen auf unvorbehandelter Biomasse wachsen und produzieren hierbei Wasserstoff und andere Plattformchemikalien wie Acetat, Formiat, Lactat, Ethanol und Succinat als metabolische Endprodukte. Trotz des hohen biotechnologischen Potenzials sind diese Organismen und ihre Enzyme weitgehend unerforscht. So sind grundlegende Wachstumsvoraussetzungen nahezu unbekannt und Kenntnisse über ihre Stoffwechselwege und deren Regulierung sind noch begrenzt. Durch Untersuchung dieser Aspekte soll diese Arbeit den Grundstein für eine biotechnologische Anwendung von anaeroben Pilzen legen. Die Nichtverfügbarkeit anaerober Pilz-Stämme in Stammsammlungen stellt eine der Herausforderungen bei der Arbeit mit diesen Organismen da. Der erste Schritt war daher eine Isolation aus dem Kot unterschiedlicher Zootiere, was zum Erhalt von sechs verschiedenen Isolaten führte. Ein aus Alpakakot isolierter Stamm stellte sich als eine neue Pilzgattung und -art heraus, die zuvor nur als SK4-Klade von sequenzierten Umweltproben bekannt war. Interessanterweise wurde der gleiche Stamm gleichzeitig aus dem Pansen eines Mähnenspringers in Texas, USA isoliert. Die Kooperation unserer Forschergruppen mündete in einer gemeinsamen Publikation, welche Kapitel 3 bildet. Isolate von beiden Orten waren nahezu identisch. Sie bildeten mittelgroße (2-5 mm) Kolonien mit einem weißen Zentrum aus Sporangien auf Agar und einen Biofilm aus Myzel in flüssigem Medium. Mikroskopisch waren monozentrische Thalli und sphärische poly-flagellierte Zoosporen mit 7-20 Flagellen zu erkennen. Sowohl die Öffnung einer apikalen Pore des Sporangiums wie auch das Aufbrechen der sporangialen Zellwand führten zur Freisetzung der Zoosporen. Dieser duale Mechanismus ist bisher einzigartig unter allen derzeit beschriebenen anaeroben Pilz-Stämmen. Ein phylogenetischer Vergleich anhand der Sequenzen der D1-D2 28S Untereinheit des rRNA Gens (LSU) und des internal transcribed spacer 1 (ITS1) wies hohe Identitätsübereinstimmungen zwischen allen Isolaten (geringste Identität jeweils 99,07 % und 96,96 %) aber geringe Übereinstimmung zu den nächsten Verwandten auf (jeweils 92,4 % und 86.7 %). Anhand der LSU konstruierte phylogenetische Bäume ordneten die Isolate als neue monophyletische Gruppe in der Orpinomyces-Neocallimastix-Pecoramyces-Feramyces-Ghazallamyces Klade ein. Sowohl die LSU wie auch die ITS1 Sequenzen der Isolate wiesen eine hohe Sequenzidentität zu Umweltproben aus Schafen und verschiedenen anderen Herbivoren auf. Interessanterweise wurden diese Proben der SK4-Klade hauptsächlich bei Tieren gefunden, die auf Sommerweiden grasten, woraus sich die Benennung der neuen Gattung, Aestipascuomyces (abgeleitet vom lateinischen Wort für „Sommerweide“), und der Spezies, A. dubliciliberans, ableitete. Die übrigen fünf Isolate konnten, wie in Kapitel 4 beschrieben, den bereits bekannten Neocallimastigomycota Gattungen und Spezies Neocallimastix cameroonii, Caecomyces spec., Orpinomyces joyonii, Pecoramyces ruminantium, und Khoyollomyces ramosus zugeordnet werden. Während der phylogenetischen Analyse wurde eine hohe Ähnlichkeit zwischen Spezies der Gattung Neocallimastix bemerkt, was zur Neuzuweisung von Neocallimastix californiae und Neocallimastix lanati zu Neocallimastix cameroonii führte. Die Daten befürworteten ebenfalls eine Neuzuweisung der Gattung Cyllamyces als Spezies der Gattung Caecomyces. Kapitel 4 beschreibt ebenfalls die biochemische Charakterisierung aller Isolate inklusive des in Kapitel 3 beschriebenen. Die Pilze wurden auf verschiedenen Kohlenstoffquellen kultiviert und die Produktion der Metabolite Wasserstoff, Acetat, Formiat, Lactat, Ethanol und Succinat, analysiert. Unter den getesteten Bedingungen produzierte Orpinomyces joyonii kein Succinat und Khoyollomyces ramosus kein Lactat. Die Ergebnisse wiesen des Weiteren auf eine sequenzielle Produktion der Metabolite unter Bevorzugung von Wasserstoff, Acetat und Formiat hin. Beim Vergleich der Metabolitproduktion während des Wachstum auf Monosacchariden und auf Stroh fiel eine höhere Wasserstoffproduktion bei letzterem auf. Dies könnte auf eine Reaktion des Metabolismus auf erhöhte Zuckerkonzentrationen hindeuten. Für das robusteste Isolat, Neocallimastix cameroonii G341, wurde ein Dunkelfermentationsprozess im Rührkessel-Bioreaktor entwickelt, was in Kapitel 5 beschrieben ist. Dazu wurden im Vorfeld weitere Optimierungen bezüglich Medienzusammensetzung und Kultivierungsbedingungen vorgenommen. Beim Testen verschiedener Stickstoffquellen führten nur Glutamin und Ammonium zum Wachstum des Isolates. Der Pilz wuchs optimal in einem Temperaturbereich von 38,5 °C bis 41,5 °C und präferierte initiale pH-Werte zwischen 6,6 und 6,8. Die höchste Wasserstoffproduktion fand jedoch bei einem initialen pH-Wert von 6,4 bis 6,6 statt. Zugabe von Wasserstoff zu Beginn des Experiments verringerte die Wasserstoffproduktion. Im Gegensatz dazu konnte durch Erhöhung des Kopfraumvolumens oder durch Agitation die Wasserstoffproduktion erhöht werden, was mit einem Abfall der gebildeten Lactat- und Ethanol-Konzentrationen einherging. Durch Kombination beider Parameter sanken Lactat- und Ethanolproduktion soweit, dass Wasserstoff, Acetat und Formiat nahezu die einzigen Produkte waren. Succinat konnte, wenn überhaupt, nur in Spuren nachgewiesen werden. Nach Etablierung des Prozesses im Rührkesselreaktor konnten die Ergebnisse aus dem Flaschenversuch bestätigt werden. Die Wasserstoffproduktion stieg mit steigender Rührgeschwindigkeit an. Während der Anstieg von 0 rpm auf 250 rpm mit einem Anstieg der Geschwindigkeit der Wasserstoffproduktion einherging, verlangsamte eine weitere Steigerung auf 600 rpm dessen Produktion. Ein ähnlicher Effekt trat beim Vergleich von Fermentationsprozessen mit und ohne pH Regulation auf. Wurde der pH-Wert konstant bei 6,8 gehalten, verdoppelte sich im direkten Vergleich zu nicht-pH regulierten Prozessen die Produktion aller Metabolite mit Ausnahme von Lactat. Diese Ergebnisse führten zu dem Schluss, dass Wasserstoff der präferierte Elektronenendakzeptor von Neocallimastix cameroonii sein könnte. In diesem Fall würden Lactat und Ethanol nur produziert, falls eine thermodynamische Beeinträchtigung der Wasserstoffbildung vorliegt. Die höchste Wasserstoffausbeute (2,406 mmol/g = 58,84 ml/g) resultierte aus Flaschenversuchen mit 5 g/l Stroh als einziger Kohlenstoffquelle mit erhöhtem Kopfraumvolumen und Agitation. Im Literaturvergleich entspricht dies ungefähr 33-50 % der für die Dunkelfermentation angegebenen Wasserstoffausbeuten auf Basis von Stroh. Allerdings wurden bei diesen Studien diese Ausbeuten nur nach einer intensiven Vorbehandlung des Strohs erzielt. Weiterhin ist der hier beschriebene Prozess mit anaeroben Pilzen noch nicht optimiert. Auch wenn das Potential anaerober Pilze, lignocellulose-haltige Biomasse effizient zu hydrolysieren, weithin bekannt ist, und eine Vielzahl ihrer Enzyme bereits heterolog exprimiert wurden, sind die wenigsten biochemisch charakterisiert. Dies betrifft besonders hemicellulolytische Enzyme, welche verglichen mit cellulolytischen Enzymen weit weniger erforscht sind. Kapitel 6 behandelt daher die Expression von Neocallimastigomycota Hemicellulasen und deren biochemische Charakterisierung. Nach ihrer Identifikation im Pilzproteom von Neocallimastix californiae konnten die GH43 Xylosidase Xyl43Nc und die GH11 Endoxylanase X11Nc in Escherichia coli exprimiert werden. Xyl43Nc wies eine hohe Aktivität gegen 4-Nitrophenol-Xylopyranosid (Km = 0,72 mM und kcat = 29,28 s-1) bei einem Temperaturoptimum von 32 °C und einem pH Optimum von 6 auf. Bei gemeinsamer Verwendung von Xyl43Nc und X11Nc konnte Xylose aus Buchenholzxylan und Weizen-Arabinoxylan freigesetzt werden. Phylogenetische Analysen von Xyl43Nc zeigten einen gemeinsamen Ursprung mit Enzymen aus Spirochaeten. Das Enzym bildete eine monophyletische Gruppe abseits der Sequenzen von Ascomyceten im phylogenetischen Baum. Hieraus ließ sich ein Ursprung des Enzyms aus horizontalem Gentransfer ableiten. Zusammengefasst legt diese Arbeit den Grundstein für eine zukünftige biotechnologische Anwendung anaerober Pilze und deren Enzyme

Value Generation in Organisations

Monograph

On the Impact of IT on Value Generating Activities in Organisations: an Ontology Based Approch.

The identification of Value Generating Activities in an organisation: the Business Model. IT resources and business processes: the OLPIT Ontology. Integrating the OLPIT with the BMO. Possible applications of the proposed approach. Limitations.The identification of Value Generating Activities in an organisation: the Business Model. IT resources and business processes: the OLPIT Ontology. Integrating the OLPIT with the BMO. Possible applications of the proposed approach. Limitations.LUISS PhD Thesi

Mycobacteria in rabbits: Searching for an efficient paratuberculosis infection model

238 p.La paratuberculosis es una enfermedad intestinal granulomatosa crónica, producida por Mycobacterium avium subsp paratuberculosis (Map) perteneciente al complejo mycobacterium avium (MAC). Esta enfermedad que afecta principalmente a los rumiantes causa cuantiosas pérdidas económicas y podría estar relacionada con la enfermedad de Crohn en humanos. El avance en el estudio de la paratuberculosis se ha visto limitado debido a la falta de un modelo de laboratorio adecuado.En esta Tesis Doctoral se ha avanzado en el estudio de la susceptibilidad del conejo a la infección por MAC, así como en la utilización del conejo como modelo animal de laboratorio, lo que ha permitido progresar en el conocimiento de la patogenia y la vacunación frente a Map. Así, se ha demostrado que la utilización de diferentes estrategias alimentarias a través de la modificación de la microbiota intestinal modula la infección por Map. Además se ha observado que la vacunación frente a Map disminuye los índices de infección a través del aumento de la respuesta inmune humoral y celular. Por otra parte, la vacunación tras el desafío ha mostrado cierto grado de efecto terapéutico. Estos hallazgos sugieren que estas estrategias deberían ensayarse con un enfoque holístico, teniendo en cuenta las posibles interacciones y/o efectos aditivos. En el presente trabajo hemos demostrado que el conejo es un modelo animal de infección de Map útil, que permite avanzar en el estudio de la infección subclínica y por ende podría ser de utilidad en el estudio de otras enfermedades inflamatorias intestinales en humanos.Neiker/Tecnali

Detection of Johne\u27s disease in an Iowa (United States) dairy herd : comparisons of the milk ELISA, serum ELISA, Gamma-Interferon and fecal culture tests and the effect of a skin-test using a cell-free sonicate of Mycobacterium avium subsp. paratuberculosis (19698) on the production of Gamma-Interferon

Analysis using kappa returned a value of -0.116 indicated a very low percentage of agreement between the two testing means. With the SELISA being the regulatory US standard it is not recommended to use the MELISA for purposes of culling decisions. Results of the skin testing portion of the study indicates that inoculation of animals with 19698 MpS significantly increased production of [Gamma]-IFN in cattle declared to have positive (MpS-P) baseline [Gamma]-IFN responses for Johne\u27s disease. Peak production of [Gamma]-IFN occurred at 144 hr following skin testing and was statistically significant P = 0.0005 as compared to day 0. Levels of [Gamma]-IFN dropped on day 9, but remained at levels that were significantly higher than day 0 (P = 0.0325). Levels of [Gamma]-IFN again significantly increased on day 27 when compared to day 15 (P = 0.0362), and was significantly higher than day 0 (P = 0.0004). Further research is warranted to determine if the addition of skin testing to the [Gamma]-IFN assay could increase test sensitivity or durability.Cattle from the Iowa State University, Ames dairy herd were characterized for Johne\u27s disease using results from the milk ELISA (MELISA) (Dairy Lab Services; Dubuque, IA), serum ELISA (SELISA) (IDEXX Laboratories, Inc., Westbrook, ME), [Gamma]-IFN assay (CSL Limited Parkville Victoria Australia) and fecal culture. Using the results of the initial [Gamma]-IFN assay and the SELISA determination, all herd members were divided into four groups consisting of: a negative SELISA and [Gamma]-IFN, suspect [Gamma]-IFN, positive [Gamma]-IFN and SELISA positive individuals. The first three groups were enrolled in the skin testing trial and were randomly assigned within group to one of two treatments. This consisted of an injection of either 100[Mu]g (0.1m1) of a M. paratuberculosis strain 19698 cell free sonicate (19698 MpS) or 0.1 ml of 0.9% saline solution as an intradermal injection. Blood samples were obtained on days 0, 3, 6, 9, 15, and 27 days post-injection for [Gamma]-IFN assay, while skin lesion measurements were made on days 0, 3, 6 and 9. The MELISA test returned the greatest number of test positive individuals at 12.96%, The SELISA was 7.41% test positive, [Gamma]-IFN assay was 4.40%, and fecal culture yielded 0% positive. Further evaluation indicated the SELISA had a significantly lower age (37.2 vs. 50.5 and 48.3 months respectively) at positive test (P\u3e [T] = 0.0192) than did MELISA or [Gamma]-IFN. Due to the low number of fecal culture and [Gamma]-IFN positive individuals, and the significantly lower age at positive test it could be suspect that the SELISA had an inordinate number of false positive individuals. MELISA and SELISA results were compared using an XY plot. No common individuals were identified as test positive by both tests

DYNAMICS OF MYCOBACTERIUM AVIUM SUBSP. PARATUBERCULOSIS IN DAIRY HERDS: INSIGHTS INTO TRANSMISSION RISKS, BULK-MILK CONTAMINATION, AND ASSOCIATIONS BETWEEN DIAGNOSTIC TESTS

Johne’s disease, a severe granulomatous enteritis of ruminant animals, is caused by Mycobacterium avium subsp. paratuberculosis (MAP). MAP infections have detrimental consequences for animal health and reduce dairy-herd productivity. Bacterial fastidiousness and slow generation time encumber diagnostic testing strategies. MAP is also a potential etiologic agent of human Crohn’s disease, with the bulk-milk supply serving as a possible transmission vector. The objective of this dissertation is to explore MAP infection dynamics on dairy farms, with an emphasis on the routes of bulk-milk contamination, transmission risk across production type, and the interplay between diagnostic testing outcomes. Accordingly, we have applied statistical and mathematical approaches to both cross-sectional and longitudinal datasets. Using questionnaire data from 292 U.S. dairies, we conducted a comparative risk assessment of organic vs. conventional management and determined that organic herds were at higher risk for new MAP infections. We concluded, empirically, that organic farms were more susceptible to a synergism of risk factors within the maternity pen and should improve calving-area hygiene if electing to permit cow-calf contact. Bulk-milk testing was also conducted for these herds. Most high ELISA tanks were PCR negative, implying that ELISA is not a perfect predictor of bulk-milk MAP status; for accurate risk assessment, bulk-milk ELISA should be used in tandem with PCR. A combination of ELISA and PCR may also aid in determining the specific route of bulk-milk contamination (either environmental or direct shedding). To extend the investigation to individual animals, longitudinal data were obtained from 14 MAP-positive cows in 2 low-prevalence herds. Robust relationships between culture, fecal qPCR, and milk ELISA were revealed, using mixed linear modeling to adjust for cow characteristics. We explored temporal relationships and observed that spikes in fecal shedding were predictive of subsequent high milk ELISA results. We also noted that disease “Progressors,” (infected animals with increasing fecal MAP CFU over time) had higher antibody titers overall. Interestingly, the paucity of positive milk samples, from both individual and bulk- tank sources, suggests that milk contamination is not a chief concern in low-prevalence herds. Armed with insights from these studies, in addition to published literature, we developed a mathematical model to explore the interaction between categories of infection, environmental MAP burden, and bulk-tank contamination. Direct shedding into milk accounted for < 1% of the MAP CFU in the tank, with environmental contamination from high shedders as the primary driver of bulk-milk MAP burden. Culling of high shedders, cleaning of the maternity pen, and adherence to milking parlor cleanliness each had a strong influence on lowering the bulk-milk MAP load. A combination of these initiatives served to drive the MAP level below an acceptable threshold (< 103 CFU/L). While complete elimination of MAP may be an unrealistic target for high-prevalence herds, the production of bulk milk with a low MAP load appears feasible. In this work, we assess the significance of a variety of contamination routes, transmission risks, and intervention strategies. These efforts are directed toward improved understanding of testing schemes and an ultimate refinement of control measures and milk quality programs. The conclusions from the studies presented in this dissertation may be applied to mitigate the spread of MAP in dairy herds, reduce prevalence, and lower or eliminate MAP in the bulk-milk supply

Perception in real and artificial insects: a robotic investigation of cricket phonotaxis

The aim of this thesis is to investigate a methodology for studying percep¬ tual systems by building artificial ones. It is proposed that useful results can be obtained from detailed robotic modelling of specific sensorimotor mechanisms in lower animals. By looking at the sensory control of behaviour in simple biological organisms, and in working robots, it is argued that proper appreciation of the physical interaction of the system with the environment and the task is essential for discovering how perceptual mechanisms function. Although links to biology, and concern with perceptual competence, are fields of growing interest in Artificial Intelligence, much of the current research fails to adequately address these issues, as the model systems being built do not represent real sensorimotor problems.By analyzing what is required for a model of a system to contribute to ex¬ plaining that system, a particular approach to modeling perceptual systems is suggested. This involves choosing an appropriate target system to model, building a system that validly represents the target with respect to a particular hypothesis, and properly evaluating the behaviour of the model system to draw conclusions about the target. The viability and potential contribution of this approach is demonstrated in the design, implementation and evaluation of a mobile robot model of a hypothesised mechanism for phonotaxis in the cricket.The result is a robot that successfully locates a specific sound source under a variety of conditions, with a range of behaviour that resembles the cricket in many ways. This provides some support for the hypothesis that the neural mechanism for phonotaxis in crickets does not involve separate processing for recognition and location of the signal, as is generally supposed. It also shows the importance of un¬ derstanding the physical interaction of the system's structure with its environment in devising and implementing perceptual systems. Both these results vindicate the proposed methodology

Normal And Epilepsy-Associated Pathologic Function Of The Dentate Gyrus

The dentate gyrus plays critical roles both in cognitive processing and in regulating propagation of pathological, synchronous activity through the limbic system. The cellular and circuit mechanisms underlying these diverse functions overlap extensively. At the cellular level, the intrinsic properties of dentate granule cells combine to make these neurons fundamentally reluctant to activate, one of their hallmark traits. At the circuit level, the dentate gyrus is one of the more heavily inhibited regions of the brain, with powerful feedforward and feedback GABAergic inhibition dominating responses to afferent activation. In pathologic states such as epilepsy, disease-associated alterations within the dentate gyrus combine to compromise this circuit’s regulatory properties, culminating in a collapse of its normal function. Through the use of dynamic circuit imaging and electrophysiological brain slice recordings, pharmacology, immunohistochemistry, and a pilocarpine model of epilepsy, I characterize the emergence of dentate granule cell firing properties during brain development and then examine how the circuit’s normal activation properties become corrupted as epilepsy develops. I find that, in the perinatal brain, dentate granule cells activate in large numbers. As animals mature, these cells become less excitable and activate in extremely sparse populations in a precise, repeatable, frequency-dependent manner. This sparse activation is mediated by local circuit inhibition and not by alterations in afferent innervation of granule cells. Later, in a pilocarpine model of epilepsy, I demonstrate that normally sparse granule cell activation is massively enhanced during both epilepsy development and expression. This augmentation in excitability is mediated primarily by local disinhibition, and the mechanistic cause of this compromised inhibitory function varies over time following epileptogenic injury. My results implicate a reduction in chloride ion extrusion as a mechanism compromising inhibitory function and contributing to granule cell hyperactivation specifically during early epilepsy development. In contrast, we demonstrate that sparse dentate granule cell activation in chronically epileptic mice is rescued by glutamine application, implicating compromised GABA synthesis as a mechanism of disinhibition in chronic epilepsy. We conclude that compromised feedforward inhibition within the local circuit is the predominant mediator of the massive dentate gyrus circuit hyperactivation evident in animals during and following epilepsy development

Development and initial validation of a dairy biological risk management assessment tool

Disease prevention protocols on dairies, either aimed at keeping disease out (biosecurity), preventing spread of disease on the farm (biocontainment), or reducing the infectious burden have always been a concern. There are a myriad of recommendations available to dairy producers to help minimize disease threats. Dairy operations differ in management style and tolerance of risk, thus there is not a one–size–fits–all answer to minimize disease entry and spread. Risks must first be identified before they can be managed. Dairy biological risk management (BRM) materials were developed to educate producers and their advisors about identifying disease risk management practices and preventing disease entry and spread to the animals in their care using the concepts of risk analysis: risk perception, risk assessment, risk management and risk communication. The BRM toolbox contains a background document reviewing published disease management protocols for dairy operations, risk management assessment questions to identify various strengths and weaknesses of disease introduction and spread, management protocols for each identified risk, and risk communication tools, all based on disease prevention through five routes of transmission (aerosol, direct contact, fomite, oral and vector-borne). The outcome was a set of peer-reviewed resources available online, free of charge, for dairy producers and their advisors to utilize. One objective of this study was to report the current biological risk management practices of California and Midwest dairies of different sizes and management styles. This was accomplished by ascertaining producer-reported prevention practices through on-farm interviews utilizing two questionnaires on 80 dairy operations in California and the Midwest. Herd size ranged from 92 to 3,550 head (average 772). There were 64 Holstein herds, seven Jersey herds, one Guernsey herd and eight mixed herds. Production (305 day mature equivalent) ranged from 15,564 to 30,586 pounds (average 24,113) and somatic cell count (SCC) ranged from 110,000 to 954,000 cells/mL (average 284,873). Reported management practices on a majority of the dairy operations included examining all feedstuffs closely for manure, mold, foreign material, and overall quality (95%), investigating animals that will not eat or do not consume all of their feed (95%), humanely and promptly euthanizing animals that are not going to recover (93.7%), keeping stalls clean (scraped at least one time daily) (92.3%), inspecting animals daily for signs of illness (90.0%), keeping alley ways clean (scraped or flushed at least one time daily) (87.5%), knowing the origin of all replacement heifers (86.3%), having a fly control program (81.3%), and regularly maintaining the dry lot area to prevent manure buildup and areas of stagnant water (80.0%). The top three responses for the biggest perceived disease risk/challenge included mastitis—all types (30 herds), FMD (11 herds), and Johne\u27s disease (nine herds). Most farms (70%) introduced animals and the highest SCC were in the herds that introduced lactating and dry cows. Very few of the herds had isolation facilities (22.5%) or utilized quarantine (22.0%) for newly introduced or returning animals. Visitors were reported to exceed 10 per week on 60% of the operations yet only 30% had any type of protocol regarding boots, animal contact, or signing a visitor log. Only 16.3% of dairies utilized their veterinarian\u27s training and skills to necropsy animals that died of undetermined causes. A majority of dairy operations (71.3%) complied with removing calves at birth prior to nursing. Only 36.3% of herds reported collecting colostrum within 2 hours of calving but nearly 74% of herds fed colostrum by six hours of age. Thirty-five herds (43.7%) in this study pooled colostrum from multiple cows; large herds (\u3e506 head) were more than twice as likely to pool colostrum as compared to smaller (\u3c505 head) herds. Scientific data that correlates management practices to production parameters is sparse. The overarching goal of this project was to identify disease prevention practices that correlated with positive outcomes on dairy operations (higher milk production, lower somatic cell count). Introducing animals to a herd did not prove significant when multiple prevention practices were included in the final model, but it remains a critical control point as an independent prevention practice for both milk production and quality. Prevention practices that correlated with higher milk production and lower somatic cell count included management styles characterized as ’attention to detail’. For instance, fly control, having a SCC less than 200,000 cells/mL, inspecting animals daily, cleaning alleyways, and preventing young animals from contacting manure from older animals were associated with higher than breed average 305 day mature equivalent milk production. The four disease prevention practices that were associated with a lower SCC included removing calves at birth prior to nursing, collecting colostrum within two hours of calving, giving a second dose (1/2 to y gallon) of colostrum 12 hours after the first feeding, and having a fly control program