La trazodone est une drogue antagoniste et agoniste dose-dépendante de la sérotonine utilisée en médecine humaine et vétérinaire pour le traitement de troubles reliés à l’anxiété. Le trauma a été identifié comme la cause primaire de morbidité et de mortalité chez plusieurs espèces de ruminants sauvages et peut être exacerbé par le stress. Dans une étude récente chez la chèvre domestique, la trazodone a réduit les niveaux d’activité sans effets secondaires. La trazodone a le potentiel de réduire le stress et de permettre des captures et manipulations plus sécuritaires chez ces espèces. Les objectifs de cette étude étaient d’identifier une dose de trazodone résultant en une diminution du niveau d’activité chez une espèce de ruminant sauvage, le gnou bleu (Connochaetes taurinus), d’évaluer la sécurité de cette dose et ses effets sur les concentrations de cortisol sérique suivant une procédure vétérinaire de routine. Une étude pilote effectuée à l’aide d’éthogrammes a permis d’identifier une dose orale de 15 mg/kg comme étant efficace pour réduire les niveaux d’activité. Cette dose a mené à une augmentation de 111% du temps passé à dormir/se reposer (P = 0.0003) et de 41% du temps à être couché (P = 0.0016) et une diminution 64% du temps passé à bouger (P = 0.005) et de 65% du temps passé à être vigilant (P = 0.026). La concentration systémique de trazodone mesurée 2 heures suivant l’administration était de 95 ± 48 µg/L. Les concentrations de cortisol sérique durant une procédure de prise sanguine n’ont pas été significativement modifiées par l’administration de trazodone (P > 0.05). Le seul effet adverse noté avec la dose de 15 mg/kg était la baisse d’appétit, ce qui n’était pas présent avec la dose de 12 mg/kg évaluée lors du projet pilote. La trazodone apparaît sécuritaire et prometteuse pour la diminution de l’activité chez le gnou bleu et ainsi pour avoir des effets positifs sur leur bien-être et la sécurité des ruminants sauvages lors de procédures vétérinaires. / Trazodone is a dose-dependant serotonin antagonist and agonist used in human and veterinary medicine to treat anxiety-related conditions. Trauma has been identified as the leading cause of morbidity and mortality in several wild ruminant species and can be exacerbated by stress. In a recent study in domestic goats, trazodone reduced activity levels without adverse effects. Trazodone could allow for safer capture and handling in ruminant species. The objectives of the present study were to identify a dose of trazodone that decreases activity levels in captive blue wildebeest (Connochaetes taurinus), to evaluate its effects on serum cortisol levels following a routine veterinary procedure, and its safety. A pilot study using ethograms identified a 15 mg/kg oral dose of trazodone as effective to reduce activity levels. Over 6 hours, this dose resulted in a 111% increase in time spent sleeping/resting (P=0.0003), a 41% increase in time spent lying down (P=0.0016), a 64% reduction in time spent moving (P=0.005), and a 65% reduction in time spent being vigilant (P=0.026). Systemic absorption of trazodone was identified when plasma concentrations were measured after 2 hours (95 ± 48 µg/L). Serum cortisol levels during a routine venipuncture event were not significantly different following trazodone adminstration (P > 0.05). Mild hyporexia was the only adverse effect noted at 15 mg/kg and was absent at a 12 mg/kg dose. Trazodone appears safe and promising to decrease activity in the blue wildebeest and might thus have a positive effect on wild ruminant welfare and the safety of veterinary procedures.

La contamination des équipements d’alimentation des veaux en pré-sevrage est une préoccupation de santé importante dans l’industrie laitière, et peut être associée à une augmentation des taux de morbidité et de mortalité. Les producteurs laitiers et intervenants ont donc besoin d’un outil pratique, rapide et fiable pour évaluer la propreté des équipements, compte tenu que l’évaluation visuelle seule est insuffisante. Les objectifs de cette étude sont de développer une méthode standardisée pour évaluer la propreté de plusieurs équipements grâce à la luminométrie, d’estimer la prévalence de contamination et décrire les pratiques de gestion recommandées pour le nettoyage. Un total de 7 troupeaux laitiers du Québec a été choisi par convenance. Après l’évaluation visuelle de l’hygiène, la propreté des équipements a été évaluée par écouvillonnage direct pour les seaux et les tétines avec des écouvillons Hygiena UltraSnap®. Une technique de rinçage a été utilisée pour les tubes à gaver, les biberons et les distributrices automatiques de lait (AMF) avec des écouvillons UltraSnap®, AquaSnap® et MicroSnap®. Pour valider la technique d’écouvillonnage direct des seaux, une étape avec le même opérateur et entre plusieurs opérateurs a été réalisée, ainsi qu’une culture bactérienne conventionnelle. Au total, 519 écouvillons ont été prélevés sur 201 équipements. La contamination médiane (intervalle interquartile) en RLU pour le biberon, le tube à gaver, l’AMF, le seau et la tétine est de 2 (1;6), 2 (0;12), 52 (19;269), 886 (128;7,230) et 899 (142;6 928), respectivement. La technique d’écouvillonnage direct, qui consiste à écouvillonner directement la surface d’un équipement, a montré une excellente corrélation pour la fidélité intra-opérateur (corrélation intra-classe (ICC) = 0,93; IC95%: 0,88-0,96). La fidélité inter-opérateur (2 sessions avec 3 opérateurs différents) a montré une corrélation élevée (ICC = 0,88; IC95%: 0,78-0,94 pour la 1ère session et ICC = 0,89; IC95%: 0,79-0,95 pour la 2e). Le score visuel (qui grade de 1 à 4 la propreté des équipements selon la quantité de matières organiques) des tubes à gaver, de l’AMF et des seaux a été positivement associé aux valeurs du luminomètre. Une corrélation positive entre la culture bactérienne et l’écouvillonnage direct des seaux a été trouvée pour l’UltraSnap (rho de Spearman (rs) = 0,653; IC95%: 0,283-0,873; P = 0,0003) et le MicroSnap (rs = 0,569; IC95%: 0,309-0,765; P = 0,002). Cette étude décrit une technique standardisée d’écouvillonnage à la ferme pour évaluer le statut hygiénique des équipements par luminométrie, qui peut être intégrée dans la gestion des problèmes de santé des veaux laitiers en pré-sevrage. / The contamination of equipment used to feed pre-weaned calves is an important health issue for the dairy industry, which can be associated with increased morbidity and mortality rates. Therefore, dairy producers and consultants need a practical, quick, and reliable tool to assess equipment cleanliness, since visual assessment alone is insufficient. The objectives of this cross-sectional study are to develop a standardized robust method to evaluate the cleanliness of several types of feeding equipment on-farm by using luminometry, with special emphasis on a direct swabbing technique to sample buckets, and to estimate the prevalence of contamination and the recommended management practices for cleaning and describing equipment. A total of 7 Quebec commercial dairy herds were selected conveniently. Following visual hygiene scoring, the cleanliness of available piece of feeding equipment was assessed using direct surface swabbing for buckets and nipples with Hygiena Ultra-Snap® swabs. A liquid rinsing technique was used for esophageal feeders, bottles, and automatic milk feeders (AMF) with UltraSnap®, AquaSnap®, and MicroSnap® swabs. To validate the direct swabbing technique of buckets, a comparison within and between operators was realized, as well as a conventional bacterial culture. A total of 519 swab samples were obtained from 201 pieces of equipment. The median (interquartile range) contamination in RLU for a bottle, esophageal feeder, AMF, bucket and nipple was 2 (1;6), 2 (0;12), 52 (19;269), 886 (128;7,230) and 899 (142;6,928), respectively. The direct swabbing technique, which consists in swabbing directly the surface of an equipment, showed an excellent correlation for inter-rater reliability (intraclass correlation (ICC) = 0.93; 95%CI: 0.88–0.96). The inter-operator (2 sessions with 3 different operators) reliability showed high correlation (ICC = 0.88; 95%CI: 0.78–0.94 for the 1st session, and ICC = 0.89; 95%CI: 0.79–0.95 for the 2nd session). The visual score (who ranks equipment cleanliness from 1 to 4 according to the quantity of organic matter) of esophageal feeders, AMF and buckets was positively associated with luminometer values. A positive correlation between bacterial culture and direct swabbing of buckets was found for the UltraSnap (Spearman’s rho (rs) = 0.653; 95%CI: 0.283–0.873; P = 0.0003) and MicroSnap (rs = 0.569, 95%CI: 0.309–0.765; P = 0.002). This study describes a standardized and practical on-farm swabbing technique for assessing the hygienic status of feeding equipment by luminometry, which can be integrated in the investigation of preweaning dairy calves’ health problems.

Echinococcus est un parasite zoonotique présent mondialement circulant entre les canidés et les rongeurs ou les cervidés. L’humain est un hôte accidentel et la maladie qu’il peut développer à la suite d’une infection peut entraîner de graves signes cliniques en l’absence de traitement. Dans la faune au Québec, on reconnaît E. canadensis, responsable de l’échinococcose kystique, alors qu’E. multilocularis, associé à l’échinococcose alvéolaire, n’a pas encore été détecté. Cependant, le diagnostic récent au Québec d’un cas humain d’échinococcose alvéolaire porte à croire que le parasite circule sur le territoire. Cette étude de type transversale menée dans 12 régions administratives au Québec d’octobre à mars 2020–2021 en collaboration avec des trappeurs a pour but d’estimer la prévalence du parasite chez les coyotes et les renards roux à proximité des zones habitées et de déterminer les principaux foyers d’infection. Un test RT-PCR sur contenu intestinal a été effectué sur 707 prélèvements à partir de carcasses récupérées par les trappeurs (423 coyotes et 284 renards roux). Au total, ce sont 38 échantillons qui sont revenus positifs à Echinococcus spp. (24 coyotes et 14 renards roux) et 25 à E. multilocularis (14 coyotes et 11 renards roux). Deux zones d’infection ont aussi été identifiées, soit en Montérégie et au Bas-St-Laurent, où le risque d’infection est de 5.4 à 14.4 fois plus élevé (p < 0.05). Des analyses de régression logistique n’ont pas permis de déterminer une association statistiquement significative entre le sexe, l’espèce, la localisation géographique et le statut d’infection. Les analyses effectuées ont permis de comparer les tests de diagnostic de coproscopie et RT-PCR pour Taenia spp. et l’identification des échinocoques. Cette étude a permis de démontrer la circulation du parasite sur le territoire québécois dans la majorité des régions administratives étudiées. / Echinococcus is a zoonotic parasite present worldwide circulating between canids as definitive hosts and rodents or cervids as intermediate hosts. Humans are accidental hosts and the disease they may develop because of an infection can lead to serious clinical signs if left untreated. In Quebec, the presence of E. canadensis, responsible of cystic echinococcosis is recognized in wildlife, but E. multilocularis has not yet been identified. The recent diagnosis of a human case in Quebec of alveolar echinococcosis, the disease caused by E. multilocularis, leads us to consider that the parasite may circulate on the territory. This cross-sectional study conducted in 12 administrative regions in Quebec from October 2020 to March 2021 in collaboration with trappers aims to estimate the prevalence of the parasite in coyotes (Canis Iatrans) and red foxes (Vulpes vulpes) near populated areas and to detect high-risk areas of infection. An RT-PCR test on intestinal contents was carried out on 707 samples from carcasses recovered by trappers (423 coyotes and 284 red foxes). A total of 38 samples were positive for Echinococcus spp. (24 coyotes and 14 red foxes) and 25 for E. multilocularis (14 coyotes and 11 red foxes). Two high-risk areas of infection have also been identified, in Montérégie and Bas-St-Laurent, where the risk of infection is 5.4 to 14.4 times higher (p < 0.05). Logistic regression analyzes failed to determine a statistically significant association between sex, species, geographic location and infection status. The analyzes allowed us to compare the diagnostic tests of coproscopy and RT-PCR for Taenia spp. and identification of Echinococcus. This study demonstrated the circulation of Echinococcus spp. and E. multilocularis on the Quebec territory in most of the administrative regions studied.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Ronette Gehring / Milk composition has a dynamic nature, and the composition varies with stage of lactation, age, breed, nutrition and health status of the udder. The changes in milk composition seem to match the changes in the expression of membrane proteins in secretory mammary epithelial cells that are needed for the movement of molecules from blood to milk and vice versa (Nouws and Ziv, 1982). Thus, an understanding of transporter expression, function and regulation in mammary epithelial cells can provide insight into mammary gland function and regulation. The goal of this project was to elucidate (molecularly and functionally) the role of drug transporters in the barrier function of an epithelial monolayer cultured from an immortalized bovine mammary epithelial cell line (BME-UV). To characterize the regulation (expression and function) of these drug transporters in BME-UV cells after exposure to cytokine TNF-α for selected periods of time. Representative members of drug transporters of the SLC (OCT and OAT) and ABC (P-glycoprotein) superfamilies were chosen for this project. In the first study, the involvement of a carrier-mediated transport system in the passage of organic cation (TEA) and anion (EsS) compounds was elucidated across the BME-UV monolayer. In the second study, molecular and functional expression of bOAT isoforms in BME-UV cells were studied. The final study characterized the effects of cytokine TNF-α on the expression and function of P-glycoprotein, an efflux pump, in BME-UV cells. Cytokine TNF-α exposure induced the expression of ABCB1 mRNA and increased P-glycoprotein production in BME-UV cells, resulting in a greater efflux of digoxin, a known P-glycoprotein substrate, back into the apical fluid. The expression, function, and regulation of these transporters in the mammary gland has important implications for understanding the barrier function of the mammary epithelium and, in more specific, for characterizing the role of these transporters in the accumulation and/or removal of specific substrates from milk and/or plasma. Moreover, this study provides an in vitro cell culture model of mammary epithelium to characterize mammary epithelial cell function during inflammation.

Doctor of Philosophy / Department of Diagnostic Medicine/Pathobiology / Steven S. Dritz / Vaccination against porcine circovirus type 2 (PCV2) has become a standard practice to improve pig mortality and growth rate in PCV2-affected herds. Unfortunately, there has been little field-based research evaluating factors which affect circovirus vaccination. The focus of this research was on potential vaccination-affecting factors such as age, dosing strategy, pig genetic makeup, and interaction with other vaccines. A total of 6,275 pigs were used to determine factors which affect circovirus vaccination and the effects of vaccination on average daily gain (ADG), immune responses, and viral circulation under field conditions. In the first study evaluating circovirus vaccination effects on PCV2 antibody titer, regardless of age and dose administration protocol, pigs vaccinated with a 2-dose circovirus vaccine had increased (P ≤ 0.008) antibody titers compared with non-vaccinates. In a second study, dosing strategy failed (P = 0.31) to affect antibody titers. However, product and time after vaccination did affect (P = 0.005) antibody titers. In another 130-d study across the nursery and finishing phases, pigs vaccinated with a 2-dose circovirus vaccine had decreased (P < 0.001) serum PCV2 viral load compared with non-vaccinates and ADG of vaccinates was better than non-vaccinates. However, the effect was more pronounced (vaccination-by-genetic interaction, P ≤ 0.05) in Duroc-based compared to Pietrain-based pigs. In a study limited to the nursery phase, vaccination for PCV2 and Mycoplasma hyopneumoniae independently reduced ADG and consumption, but the effect was product-dependent. In a 155-d study across the nursery and finishing phases, vaccination with a 2-dose, 2-vaccine program for PCV2 and Mycoplasma hyopneumoniae decreased (P < 0.001) nursery ADG but tended to increase (P = 0.06) finishing ADG compared to a 1-dose, 2-vaccine program, with no difference (P = 0.66) observed between final pig weights. Finally, circovirus vaccination affected PCV2-circulation in high-health research herds but not in a commercial herd where PCV2 DNA was detected in the environment. These results indicate that finishing performance was improved by a 2-dose circovirus vaccine; however, nursery performance was negatively affected by the same product. Circovirus vaccination responses of growth, viral load, and antibody titer were affected by pig genetic makeup, product, and PCV2-exposure status.

Master of Science / Department of Clinical Sciences / David E. Anderson / Drug delivery systems for time release of recombinant human bone morphogenetic protein-2 (rhBMP-2) and antibiotics in orthopedic surgeries continue to be developed. Recently, a biodegradable novel polymeric matrix has been developed for this purpose. We hypothesized that impregnation of the matrix with rhBMP-2 would enhance bone healing. The objectives of the study were to characterize elution of rhBMP-2 and two antimicrobials (tigecycline, tobramycin) from the matrix, and bone response to the matrix in the presence or absence of rhBMP-2 and antimicrobials. In vitro elution of tigecycline, tobramycin, and rhBMP-2 from the matrix was investigated. Drug concentration in media were measured on days 1-6, 8, 10, 13, 15, 17, 21, 25, 28, and 30 using high pressure liquid chromatography/mass spectrometry/mass spectrometry (HPLC/MS/MS; antimicrobials) and ELISA (rhBMP-2). In vivo testing was done using a unicortical defect created into each tibia of twenty adult goats. Animals were randomly assigned to one of 5 groups: 1) control (untreated defect); 2) matrix; 3) matrix+ antimicrobials (tigecycline+tobramycin); 4) matrix+rhBMP-2; and 5) matrix+antimicrobials+rhBMP-2. Plasma concentration of tigecycline and tobramycin and serum concentration of rhBMP-2 were measured by the above techniques on days 1-7, 9, 11, 13, 15, 17, 22, 26, and 30. Bone response was assessed on days 0, 14, and 30 using radiographic scoring and dual energy X-ray absorptiometry (bone mineral density [BMD]). After euthanasia on day 30, histomorphologic analyses of the bone defects were done. Categorical variables were analyzed using a generalized linear model, and continuous variables using an ANOVA with P < 0.05 considered significant. In vitro elution was characterized by a rapid release on day 1 followed by a slow release until day 30 for both antimicrobials and rhBMP-2. Plasma antimicrobial concentrations showed continued release throughout the study period. Serum rhBMP-2 concentration, radiographic scores and BMD were not significantly different between groups. Periosteal and endosteal reaction surface areas were significantly greater surrounding the defects in group 4 (matrix+rhBMP-2). There was no significant difference between the groups for the percent of bone filling the defect. The matrix served as an appropriate antimicrobial and rhBMP-2 delivery system and successfully stimulated bone production when rhBMP-2 was present.

Master of Science / Department of Diagnostic Medicine/Pathobiology / Roman Reddy R. Ganta / Ehrlichia and Anaplasma species are rickettsial organisms which infect a variety of mammalian species. The organisms are transmitted from ticks and are maintained in reservoir hosts. Several pathogens have been identified in recent years as the causative agents for emerging infections in people. One of the primary reservoir hosts for the pathogens is the white tailed deer. In this study, 147 deer blood samples and 37 ticks were evaluated for the prevalence of Ehrlichia/Anaplasma species by TaqMan-based real time amplification assay and DNA sequence analysis. One hundred and thirteen (74%) samples tested positive with the Ehrlichia/Anaplasma genera-specific probe. Further analysis of the samples with the probes specific for human ehrlichiosis agents, E. chaffeensis and E. ewingii identified 4 (2.7%) and 7 (4.7%) positives, respectively. Test positives from 24 randomly selected samples were further evaluated by sequence analysis targeting to a 450 bp segment of 16S rRNA gene. All 24 samples were confirmed as positive for the Ehrlichia GA isolate # 4 (GenBank #U27104.1). DNAs from 37 pools of ticks collected from the white tailed deer were also evaluated. The TaqMan-based real time PCR assay with Anaplasma/Ehrlichia common probe identified 29 (78%) tick pools as positives whereas E. chaffeensis- and E. ewingii-specific probes identified three (8%) and one (3%) positives, respectively. The PCR and sequence analysis of tick samples identified Gram-negative bacteria species which included one endosymbiont of Rickettsia species (one tick pool), one Alcaligenes faecalis strain (three tick pools), five different Pseudomonas species (9 tick pools) and five different uncultured bacteria organisms (7 tick pools). Although the pathogenic potential of the white-tailed deer isolates of Anaplasma and Ehrlichia agents remains to be established, their high prevalence and the presence of human ehrlichiosis pathogens in white-tailed deer is similar to earlier findings. The high prevalence of the deer isolates of Anaplasma and Ehrlichia species demonstrates the need for further assessment of the pathogenic potential of these organisms to people and domestic animals.

Doctor of Philosophy / Department of Diagnostic Medicine and Pathobiology / Raymond R. R. Rowland / Porcine Reproductive and Respiratory Syndrome Virus (PRRSV) is the most costly virus to the swine industry, worldwide. This study explored the application of deep sequencing techniques to understand better the virus-host interaction. On the virus side, PRRSV exists as a quasispecies. The first application of deep sequencing was to investigate amino acid substitutions in hypervariable regions during acute infection and after virus rebound. The appearance and disappearance of mutations, especially the generation of a new N-glycosylation site in GP5, indicated they are likely the result of immune selection. The second application of deep sequencing was to investigate the quasispecies makeup in pigs with severe combined immunodeficiency (SCID) that lack B and T cells. The results showed the same pattern of amino acid substitutions in SCID and normal littermates and no different mutations were identified between SCID and normal littermates. This suggests the mutations that appear during the early stages of infection are the product of the virus becoming adapted to replication in pigs. The third application of deep sequencing was to investigate the locations of recombination events between GFP-expressing PRRSV infectious clones. The results identified different cross-over occurred within three conserved regions between EGFP and GFPm genes. And finally, the fourth goal was applied to develop a set of sequencing tools for analyzing the host antibody repertoire. A simple method was developed to amplify swine VDJ repertoires. Shared and abundant VDJ sequences that are likely expressed by PRRSV-activated B cells were determined in pigs that had different neutralization activities. These sequences are potentially correlated with different antibody responses.

Doctor of Philosophy / Department of Anatomy and Physiology / Chris R. Ross / The immediate antiviral defense residing in the innate immune system of multicellular organisms critically determines the outcome of viral infection. This dissertation presents a study of the "effectors" and "receptors" of porcine innate immunity in infection caused by porcine reproductive and respiratory syndrome virus (PRRSV), which is the most devastating pathogen impacting the swine industry. In the first investigation, eleven novel porcine host defense peptides (HDPs), [Beta]-defensins (pBDs), were identified and characterized. All of these peptides have a consensus [Beta]-defensin motif and phylogenetically are similar to orthologs from other species. A differential expression pattern for these 11 newly identified genes was found. For example, pBD-2 and pBD-3 were expressed in bone marrow, lung, skin and other lymphoid tissues. pBD-2 and pBD-3 were further characterized for their gene structure, and antimicrobial activity of synthetic peptides. The second study was conducted to evaluate PRRSV-induced differential expression of porcine HDPs and direct antiviral activity of selected HDPs against PRRSV. In vitro incubation of PRRSV with synthetic pBD-3 or protegrin-4 (PG-4) significantly inhibited viral infectivity. Using nine protegrin-derived peptides, it was determined that cyclization of PG-4 increased anti-PRRSV activity and mutation of some residues in PG-4 diminished some of the activity. These findings suggest the potential role of porcine HDPs as a group of innate antiviral effectors. In the third and fourth investigations, porcine Toll-like receptor (TLR) 3 and TLR7 were identified and functionally expressed. Increased expression of TLR3 was observed in PRRSV-infected porcine lungs. Stimulation of porcine alovelar macrophages with poly (I:C), a synthetic TLR3 ligand, increased expression of interferon-[Beta] and suppressed PRRSV infectivity. Activation of porcine TLR3 overexpressed in a PRRSV-sensitive cell line, elicited antiviral responses to PRRSV infection. Partial silencing of TLR3 in PAMs resulted in increased PRRSV infection. In summary, these data provide molecular information on porcine TLR3 and TLR7, and their involvement in PRRSV pathogenesis, which may elicit new strategies to prevent this costly swine disease.

Master of Science / Department of Anatomy and Physiology / Bruce D. Schultz / Epithelial cells lining the vas deferens modify the luminal contents to which sperm are exposed in response to neuroendocrine, autocrine and lumicrine transmitters. The role and identity of vas deferens epithelial potassium channels that provide the correct luminal environment for sperm maturation and delivery have not yet been determined. Cultures of vas deferens epithelial cells isolated from adult pigs were employed to investigate contributions of selected ion channels to net flux. A two-pore potassium channel, TASK-2, was identified on the apical membrane of cultured primary porcine vas deferens epithelial cells (1°PVD). Bupivacaine, a known TASK-2 inhibitor, when added to the apical bathing solution, inhibited forskolin- stimulated short circuit current, Isc, in a concentration dependent manner with a maximum inhibition of 72 ± 6% and an IC50 of 7.4 ± 2.2 µM. Apical exposure of 1°PVD cells to quinidine, lidocaine, and clofilium (other known TASK-2 blockers) inhibited forskolin-stimulated Isc in a concentration dependent manner. Fitting a modified Michalis-Menten function to the data revealed IC50 values of 274 µM, 531 µM, and 925 µM, respectively. Riluzole, a two-pore potassium channel activator, stimulated bupivacaine-sensitive Isc, further confirming the contribution of TASK-2 to net ion flux. Western blotting demonstrated the presence of TASK-2 immunoreactivity in 1°PVD cell lysates, while immunocytochemistry demonstrated apical localization of the targeted epitope in virtually all cells lining native porcine vas deferens. These results suggest that TASK-2 likely plays a role in vas deferens epithelial ion transport that may account for the reportedly high concentration of potassium in the male reproductive duct lumen. TASK-2 likely contributes to male fertility as an integral member of the regulated transport processes that account for the luminal environment to which sperm are exposed.