Brief Summary of Minutes of Annual Meeting

NE-1028 Multi State Research Project Annual Meeting Mastitis Resistance to Enhance Dairy Food Safety Annual Business Meeting Chicago, Illinois USA November 2, 2011 1. The 2011 business meeting of the NE 1028 Multistate research project was called to order by vice-chair Anne Lichtenwalner, presiding in the absence of the current chair Christina Petersson-Wolfe. 2. Christina Petersson-Wolfe distributed the meeting agenda by email prior to the meeting and during the meeting there was unanimous agreement to proceed with the proposed agenda. 3. An overview of the2010 business meeting minutes was presented (the complete minutes of the 2010 business meeting were sent by email and available on the project web site). A motion for approval was seconded and the 2010 business meeting minutes were approved by unanimous vote. 4. Introduction of members and new members: Attending members introduced themselves. Members present: 12 stations represented Tanya Gressley  University of Delaware Pedram Rezamand  University of Idaho Leo Timms  Iowa State University Bill Owens  Louisiana State University Anne Lichtenwalner  University of Maine Ron Erskine  Michigan State University Sandra Godden  University of Minnesota John Middleton  University of Missouri Dave Wilson  Utah State University John Barlow - University of Vermont Larry Fox  Washington State University Pamela Ruegg  University of Wisconsin 5. Administrative Advisor: Dr. Cameron Faustman was unable to attend due to scheduling conflicts. 6. USDA-CSREES advisor comments: Dr. Gary Sherman, the current advisor of record, was unable to attend due to scheduling conflicts. 7. Report on Mastitis Research Workers (MRW) meeting 2010 in Atlanta: 36 research talks were given with 42 attendees. Members representing 17 stations were present at the business meeting Nov. 4, 2010. This year there were 12 stations represented at the business meeting on Nov. 2, 2011. There were 27 research presentations and 30 attending MRW from Nov. 1 to 3, 2011. 8. Multistate projects: Station reports are due on November 11. A concise report is due from each participant. All participants are encouraged to report if they are not receiving emails as there may be some recent issues with server filtering emails. 9. NE 1028 Proposal re-write: The proposal rewrite is due March 2012 for the next 5 year period. The steering committee is Christina Petersson-Wolfe, Anne Lichtenwalner, and David Kerr. Individual stations will need to provide a report summarizing 5-year summary of reportable scholarly outputs and how work aligns with current objectives. Participants should also outline a 5 year plan of work and how this proposed work aligns for current objectives, and participants should give thought to revision of current proposal objectives for the rewrite. Examples of potential areas to include when revising objectives were issues of animal well-being and strategies which focus on mastitis prevention in order to reduced antimicrobial use. It was suggested that the MASCAP proposal would be an excellent source of material for the project rewrite. Draft of the rewrite will be sent to all members. 10. NE 1028 Termination report: A project termination report will also be due. Again all stations will need to provide a 5 year summary of work (publications and presentations) aligned to the relevant objectives. Annual summary reports can be used to construct the termination report. 11. Collaborative research projects: Project members participated in conference calls regarding the development of collaborative research proposals for the USDA-AFRI RFP Extension driven disease prevention and control in animals. Dr. Peter Johnson participated in one conference call providing members with information on the RFP. 12. Election of new members: Tanya Gressley was recognized as a new member and as already being listed in Appendix E. It was noted that Dr. Gressley needs to be added to the project email list. 13. Election of New Officers: Pradam Rezamond was nominated for Secretary, the nomination was seconded and all attending voted in favor. 14. Next meeting: The timing and location of the next meeting was discussed. Chicago and Minneapolis were proposed as possible meeting locations for next year. There was discussion regarding the location and ease of travel for participants and the increasing cost of meeting facilities at the current location. It was agreed that a vote on the location would be conducted by email. Bill Owens and Sandra Godden will research lodging and facilities fees for the current Chicago and possible Minneapolis locations and this information will be sent with the request for voting by email. The email vote was distributed on Nov. 9, and Chicago was selected by a majority for the 2012 meeting. MRW will be from Nov. 6 to 8 (same format as this year), with the 2012 NE 1028 business meeting on Nov. 7. 15. 2013 meeting  Ghent, Belgium was proposed as the location for the 2013 MRW meeting location. An email vote was conducted and a strong margin voted in favor of Ghent as the location for the 2013 MRW meeting. 16. Seeing no other business the meeting was adjourned after a motion and second. Respectfully submitted by John Barlow, acting as secretary for David Kerr.

Objective 1: Characterization of host mechanisms associated with mastitis susceptibility and resistance. DELAWARE The goal of this study is to evaluate the effect of thymol on function of bovine neutrophils. Thymol is a natural plant-derived compound with antimicrobial and antioxidant activities that may provide an alternative to traditional antibiotics for treatment of mastitis. The antimicrobial activities of thymol may assist neutrophils in destroying pathogens and the antioxidant activities may reduce mammary tissue damage from reactive oxygen species released by neutrophils. IDAHO We are investigating the effect of retinol-binding protein (RBP) status (as regulated by dietary retinol and/or protein) on IMI in periparturient Holstein cows. We have demonstrated a strong positive correlation between mRNA expression of TNF-alpha and that of RBP in the liver and adipose tissues. However, a follow up study showed the relationship between RBP and TNF-alpha is not a cause-effect relationship under our experimental condition. GEORGIA Immune response of dairy heifers and dairy goats to vaccination against Staphylococcus aureus was evaluated at a university dairy herd and a commercial goat herd. Results demonstrated that vaccination reduced new staphylococcal infections, herd somatic cell counts, and culling rates in dairy goats. Daily feeding of a general immunostimulant to dairy heifers resulted in increased serum L-selectin and interleukin-8 levels, increased phagocytic ability of neutrophils and monocytes against S. aureus, which may enhance immunity in heifers vaccinated against mastitis caused by this mammary pathogen. MICHIGAN Continued research has investigated the impact of excessive lipid mobilization and altered adipose tissue function during the periparturient period of dairy cows on susceptibility to mastitis and other inflammatory based diseases. Particular interest has focused on the role of oxidative stress in the initiation and expression of inflammatory pathways. Michigan will continue to investigate lipid mobilization and adipose tissue function on mastitis and other inflammatory diseases. NEW YORK The Cornell team worked on characterization of immune response after intramammary E. coli challenge in late gestation. Immune response profiles showed a predominance of Th2 biased immune response. OHIO The development of endotoxin tolerance in mid-lactation Holstein cows that experience chronic subacute ruminal acidosis or acute and subsequent endotoxin mastitis was investigated in the current trial. Cows that experienced subacute ruminal acidosis and subsequent experimental endotoxin mastitis experience endotoxin tolerance at the level of the mammary gland, but not a systemic reduction in inflammatory mediators. TENNESSEE The CXCR1 gene is highly polymorphic with more than 30 variations, 5 of which cause amino acid changes, 4 which have the potential to influence receptor function. A common naming system utilizing HGVS guidelines was suggested for this gene. Closer examination of the CXCR1c.+735 SNP revealed that neutrophils from cows with a CC genotype have lower F-actin expression and do not travel directly towards IL8. Each of these effects has the potential to influence neutrophil migration and subsequent mastitis resistance. We also have identified that CXCR1 is expressed by mammary epithelial cells under normal and infected conditions. UTAH Contributing author to invited Consensus Statement for the American College of Veterinary Internal Medicine on Mycoplasma bovis infections in cattle. VERMONT Experiments are being conducted using an ex-vivo fibroblast model to evaluate the hypothesis that polymorphisms in genes associated with pathogen detection and intracellular signaling pathways contribute to differences between cows in their innate resistance to mastitis. In vitro responses of the fibroblasts to LPS reveal substantial between-animal variation in IL-8 production allowing classification of cows as high and low responders. This variation appeared to correlate with longer duration of somatic cell count elevation in the high vs. low responding animals following in vivo E. coli challenge. VIRGINIA Our characterization of dendritic cells in the bovine mammary gland were recently published and are the stepping stone to our current work. Currently we are evaluating the immune response of bovine T cells following activation by S. aureus infected dendritic cells. Understanding of the immune activation and suppression during this infection is the only means of identifying novel therapeutics. These studies also allow us to evaluate the S. aureus antigens presented by dendritic cells using proteomic technologies. Objective 2: Characterization and manipulation of virulence factors of mastitis pathogens for enhancing host defenses. GEORGIA Assessment of 46 non-hemolytic staphylococcal isolates from heifers and cows in 2 herds, presumptively identified as coagulase-negative staphylococci (CNS) on blood agar demonstrated that 40% were actually S. aureus based on coagulase production and fermentation of mannitol on mannitol salt agar, 17% were other coagulase-positive staphylococci (CPS), and 43% were CNS. Thus, a significant proportion of S. aureus and CPS infections may be misdiagnosed as CNS, and additional testing should be performed to correctly identify such staphylococcal isolates. Characteristics common to such isolates included a somatic cell count >1 x 106/ml; off-white to gray colonies; an indistinct, narrow zone of clear hemolysis; raised center in colony; and greening/browning of the agar. IOWA Clinical mastitis samples that resulted in "No Growth" utilizing traditional culture techniques and comparative normal quarter milk samples from the same cow were tested utilizing PCR amplification 16 S rRNA genes and sequencing the hypervariable V2-V3 region to identify and quantify all bacterial species in the samples. Our metagenomics analysis shows a distinctly different and very diverse bacterial flora from normal quarters compared to culture negative clinical mastitis samples on the same cow. KENTUCKY A MS student in Microbiology at Eastern Kentucky University (Erica Lynch) is conducting a project to assess antimicrobial resistance in pathogens responsible for causing bovine mastitis in Kentucky. This project is a field survey of multiple herds in Kentucky. LOUISIANA Hill Farm is conducting antimicrobial susceptibility testing on mastitis pathogens to monitor resistance levels. Comparisons will be made between mastitis pathogens, poultry pathogens and human pathogens. A program for monitoring all herds in the state for mycoplasma mastitis continues. MAINE Pasteurization trials of Maine prototheca isolates were conducted to evaluate the sensitivity of these P. zopfii genotype II strains to standard pasteurization methods, and to explore the mechanisms for this resistance. MICHIGAN Michigan has evaluatee the application of RT-qPCR for Mycoplasma bovis detection in milk to herd mastitis programs, to be used as a method of surveillance of herd prevalence. MINNESOTA Farm prevalence of S. aureus, including MRSA, was estimated from bulk tank milk sampled in duplicate over 3 seasons from 50 MN dairy herds. The bulk tank level prevalence of S. aureus and MRSA was 62% and 1.3%, respectively. Antibiotic susceptibility testing revealed that 17.2 % (16 of 93) of S. aureus isolates were multi-drug resistant (MDR), and confirmed the presence of 2 MRSA isolates. Seven isolates, including the two MRSA isolates, produced staphylococcal enterotoxins B, C, D and E on overnight culture in nutrient media. These results suggest that MRSA genotypes associated with hospitals and community are present in MN bulk tank milk, albeit at a very low prevalence. MISSOURI We are continuing to work on the relationship between coagulase negative staphylococcal (CNS) intramammary infection (IMI), milk somatic cell count, and duration of infection. Recently we have begun mapping the genome of 4 CNS species isolated from milk. We are completing studies on risk of IMI in heifers at calving based on pre-partum IMI. NEW YORK The Cornell Team worked on characterization of the pathogenicity of gram-negative coliforms, particularly E. coli and Klebsiella species. Bacterial survival in dry cow secretum was an important predictor of pathogenicity in early lactation. TENNESSEE Tennessee is conducting further validation of the potential for Streptococcus uberis Adhesion Molecule (SUAM) as a realistic vaccine candidate. This includes: predicted antigenic regions of SUAM are involved in adherence to mammary epithelial cells, mutation of the SUA gene reduces adherence to mammary epithelial cells, and antibodies generated against SUAM protect against adhesion in vitro. Also, a mutant library was generated from a Streptococcus uberis strain UT888, which has been partially screened for genes tied to mammary adhesion and evasion of neutrophil killing. Several candidates have been identified. The intracellular fate of chronic vs acute strains of E. coli was evaluated in vitro. (TN, NY) VIRGINIA We are currently using proteomic technology, stable isotope labeling with amino acids in cell culture (SILAC), to compare the expression profiles of commensal and pathogenic strains of Staphylococcus aureus. Our goal is to identify differentially expressed virulence factors that can be targeted to prevent bacterial growth during infection. Objective 3. Assessment and application of new technologies that advance mastitis control, milk quality and dairy food safety. CONNECTICUT A study was conducted to determine the sensitivity and detection limits of the Delvotest® SP-NT and the IDEXX ² Lactam SNAP test for detection of natural antimicrobial compounds in milk and determine if pasteurization of milk containing the compounds affected the sensitivity rates. The IDEXX ² Lactam SNAP test did not detect any of the compounds evaluated. The Delvotest® SP-NT detected caprylic acid, the plant-derived antimicrobials, and nisin at their MIC values with sensitivities of 1.0. The sensitivity rates of the Delvotest® SP-NT for the detection limit (DL) ranged from 0.93 to 1.0 for the compounds evaluated. Although the MIC could not be established for phyto-mast® in this study, the Delvotest® SP-NT detected phyto-mast® at a concentration of 12.5% in milk. Nisin was the most sensitive compound with a DL 99.8% less than its MIC. Trans -cinnamaldehyde was the most sensitive of the plant-derived essential oils. The pasteurization of milk did not have an effect on the outcome of the Delvotest® SP-NT. The Delvotest® SP-NT detected the combination of eugenol and trans-cinnamaldehyde and eugenol and thymol at their respective concentrations below the DL. GEORGIA Evaluation of a non-lactating cow antimicrobial intramammary infusion product (Spectramast DC) against clinical mastitis caused by Streptococcus uberis was conducted at 2 UGA dairies. Treatment of 92 clinically infected quarters of 46 heifers (2 quarters/heifer) approximately 45 days prior to expected parturition resulted in a 100% cure rate. Additionally, all animals calved free of antimicrobial residues with low SCC and normal milk production. IOWA 1) A trial was completed evaluating maintaining effective cephapirin antibiotic levels as well as adherence to label withdrawal times and residue levels when cows were milked 3X and treated once daily in all 4 quarters for 8 days (extended therapy). Effective cephapirin levels were maintained and antibiotic withdrawal times met label specifications. 2) 23 studies evaluating teat health and condition using 15 different lactating cow teat dips and compounds were completed. 3 new teat dips were made commercial with our data on skin conditioning being the final piece with 3 new, novel dips and compounds pending. These projects included 5 different industry partners. 3) A non-inferiority study examining the effects of 3 different dry cow antibiotics and their effects on prevention and cure of IMI across the dry period, calving, and early lactation, and clinical mastitis and production through 100 days in milk was initiated (in conjunction with MN and CA). 4) 24 new dry cow teat dip sealant prototypes were evaluated for teat protection persistency compared to available commercial industry dry cow teat dip sealants in a randomized within udder model (4 dips simultaneously randomized within cows across quarters). All dips showed lower average persistency times and higher % of teats unprotected at 72 hours. KENTUCKY A survey of Kentucky herds with annual SCC <250,000 was conducted in 2011 to identify management practices associated with low somatic cell count. An economic dashboard decision support tool was developed to help dairy producers understand the economic impact of lowering SCC (www2.ca.uky.edu/afsdairy/MilkQualityCalculator). A field survey of compost bedded pack barns was conducted including bacterial analysis of bedding materials. MAINE We continue to apply the nested PCR developed to detect prototheca in bulk tank filters as a screening tool. We continue to provide mastitis detection and treatment information to Maine farms, and to interact with Extension colleagues to solve mastitis-related farm problems. MINNESOTA 1. An observational study, conducted to assess the effect of sampling interval and variation of the BMSCC data on the accuracy to predict BMSCC, concluded that the length of the sampling interval greatly influences the usefulness of BMSCC data to monitor trends in udder health at the herd level. 2. An observational study, designed to investigate the relationship between herd SCC level and performance rank for mastitis and milk quality benchmarking on dairy farms, found that herd SCC was associated with the percent infected, percent of current cows with new infections, percent of fresh cows with chronic infections and percent infected less than 30 days in milk. 3. An observational study, designed to investigate the chemical and bacteriological characteristics of digested, composted, and separated raw manure solids in Midwest dairy herds, reported that anaerobic digestion of manure may affect the characteristics of recycled solids prior to use as bedding for freestalls. 4. An observational study, designed to evaluate mortality and herd turnover rates in Midwest dairy herds utilizing recycled manure solids in deep bedded or mattress based freestalls, reported that stall surface had a relatively minor association with mortality and turnover rates in dairy herds utilizing recycled manure solids as bedding. 5. An observational study, designed to identify and quantify the months where herds have the highest percent of fresh cows with new infections (FNI) and percent of current lactating cows with new infections (CNI), reported that the greatest challenge for FNI comes during the spring when cows may be exposed to the dampest environments. The summer months are the worst for CNI, with the addition of the deep winter months for the poorest milk quality producers. 6. A randomized multistate clinical trial reported that use of an on-farm culture system to guide the strategic treatment of clinical mastitis reduced intramammary antibiotic use by half and tended to reduce milk withholding time by one day, without significant differences in days to clinical cure or bacteriological cure risk, and no differences in long-term outcomes such as recurrence of clinical mastitis in the same quarter, somatic cell count, milk production, and cow survival for the rest of the lactation after clinical mastitis. (Project completed between UMN, UWI and U of Guelph) 7. We propose the development of two parallel curriculums for web delivery to veterinary audiences: i) Mastitis: biology and economics, control, treatment, pharmacology, regulatory issues, milk quality protection. We will also seek out suitable on-line opportunities for people who wish to learn more about fundamental lactation biology, and ii) Milking system function and analysis. The project will develop a series of short content modules with appropriate text, audio, pictures, and video that would allow an online user to work through the curriculum one small piece at a time. Work will be completed in 2011 and 2012. 8. This project is a multi-state, multi-herd randomized clinical trial designed to compare the risk for presence of an IMI after calving, the risk for cure of an IMI during the dry period, the risk for developing a new IMI during the dry period, and lactation udder health and performance in the first 100 days of lactation, for quarters and cows previously treated at dry off with ToMORROW® Dry Cow, QUARTERMASTER®, or SPECTRAMAST® DC at dry off.. Cow enrollment and follow-up will be completed in 2011. (Completed in cooperation between UMN, ISU and UC-Davis) MISSOURI We are continuing to evaluate internal teat sealants during the dry period. To date we have developed radiographic and ultrasonographic methods for documenting the presence of sealants within the teat cistern during the early dry period. We have evaluated several milker hand hygiene protocols and teat end aseptic preparation techniques. NEW YORK The Cornell team worked on Listeria monocytogenes contamination of raw milk. A risk assessment was developed as well as risk factors for Listeria persistence in the milking equipment. TENNESSEE The Tennessee team continued work on evaluation of bulk tank milk quality. UTAH Multiple Mycoplasma spp. were detected in bulk tank milk samples using real-time PCR, including the first report of M. gateae in bovine milk. Results were also compared with conventional mycoplasma milk culture methods. VERMONT A split herd negative controlled field trial was conducted to evaluate the direct and indirect effects of treating chronic subclinical Staphylococcus aureus mastitis. Pulsed-field gel electrophoresis (PFGE) and multilocus sequence typing (MLST) were used to determine strain specific infection dynamics in treated and control groups. Results of this study provide strain specific estimates of the transmission parameter for S. aureus MLST clonal complexes in 2 commercial dairy herds, and evidence that treatment of subclinical S. aureus mastitis during lactation may impact transmission dynamics. VIRGINIA We collected data to evaluate the use of a non-steroidal anti-inflammatory drug (NSAID) during E. coli mastitis. NSAID administration alleviated the negative impacts on feed intake and also milk production. Animal activity was not affected by NSAID administration but was significantly altered when compared to a negative control. Data has been collected to examine the changes in animal activity around dystocia, subclinical ketosis, clinical mastitis, and milk fever in the transition period. Our results show that on the day of calving, rest bouts increased in animals that experienced dystocia over those who did not experience dystocia. Further, cows experiencing subclinical ketosis displayed increased rest bouts on the day prior to disease and decreased daily steps 6 days before the disease was diagnosed. Additionally, cows experiencing clinical mastitis had decreased rest times beginning 3 days prior to the onset of disease as compared to animals without mastitis. Cows with milk fever displayed more rest bouts with decreased daily steps on the day prior to and the day after disease diagnosis and increased overall rest duration and time after the clinical diagnosis of disease compared to cows that were not diseased. WASHINGTON We hypothesized that teat disinfection would reduce the incidence of IMI by CNS, but the reduction would not be uniformly observed in all species. Findings supported the hypothesis as Staphylococcus chromogenes and Staphylococcus xylosus were the most prevalent CNS causing IMI without disinfection; but with disinfection prevalence of S. xylosus IMI was reduced while S. chromogenes IMI was unchanged. In aggregate, post-milking teat disinfection reduced teat canal colonization and IMI by CNS. The purpose of this study was to develop real-time PCR assays to speciate three common mycoplasma mastitis agents: M. bovis, M. californicum and M. bovigenitalium. Partial 16S rRNA gene sequencing of isolates was used as the gold standard to determine agreement with the real-time PCR assay. To ascertain the specificity of the real-time PCR assay, reference strains of Mycoplasma species, Acholeplasma axanthum and common gram positive and negative mastitis pathogens were tested. No cross-reactivity of the developed real-time PCR assays with M. bovis, M. californicum and M. bovigenitalium and these reference strains were observed. Mycoplasma isolates from milk samples (n = 228) and other organ sites (n = 40) were tested by the developed real-time PCR assays and the gold standard assay. The novel real-time PCR assays is a promising diagnostic technique that may be used either as a screening or a confirmation test for mycoplasma mastitis. WORK IN PROGRESS OBJECTIVE 1: Characterization of host mechanisms associated with mastitis susceptibility and resistance. CONNECTICUT A study is planned to investigate the effect of antibiotic use against mastitis on the occurrence of antibiotic resistance in dairy farm manure. This study is a collaboration between The University of Connecticut and Yale University, Biological and Medical Sciences. DELAWARE Delaware will complete experimental work by evaluating effect of thymol on bactericidal function of neutrophils. GEORGIA Study of the immune response of dairy cows to vaccination against Staphylococcus aureus will continue. Heifers vaccinated against S. aureus and receiving an oral immunostimulant daily will be challenged with S. aureus to determine if they are more resistant to challenge compared with unvaccinated control heifers receiving the oral immunostimulant. IDAHO Idaho will continue conducting a study entitled  Interaction among energy status, retinol-binding protein status and intra-mammary infection in periparturient dairy cows. Over the past ~ 11 months 56 multiparous periparturient Holstein cows have completed the feeding trial and necessary samples have been obtained (blood, milk, PMN for killing assay, PBMC for gene expression studies, feed, liver, adipose tissue). We anticipate that completion of this project will take ~ 2 yr from now. We finished testing the effect of pro-inflammatory mediators on hepatic tissues mRNA expression of retinol-binding protein and now are in the process of establishing gene expression of RBP during the periparturient period. MICHIGAN Michigan will continue to investigate lipid mobilization and adipose tissue function on mastitis and other inflammatory diseases. MISSOURI In the coming year we will be evaluating a novel therapeutic regime for chronic Staphylococcus aureus IMI. Our heifer mastitis project evaluating risk of IMI post-calving will be completed. NEW YORK We will continue our work on immune profiling the response to an intramammary challenge in late gestation. Cellular immune mechanisms will be studied in more detail. We will also be evaluating immune response profiles in mid lactation using the same challenge model as the one we use in late gestation. TENNESSEE We will continue to assess the role of CXCR1 and its sister receptor CXCR2 in relation to immune function and subsequent resistance to mastitis. VERMONT We will evaluate the between-animal variation in the innate immune response of dermal fibroblasts from 50 Holstein cows and then challenge the four lowest and four highest ranked animals with Staphylococcus aureus to determine the relationship between in vitro ranking and in vivo mastitis resistance. VIRGINIA In the upcoming year we will continue to collect data on the immune activation and suppression during S. aureus infection. Our goal is to define the pathways regulated by this pathogen so that we can therapeutically manipulate them to change the course of infection. WASHINGTON The effect of dexamethasone induced alterations of the bovine immune system and associated mycoplasma colonization and infection will be studied. Calves will be split into treatment (dexamethasone treatment) and control. Pre-weaned calves will be challenged with M. bovis, a wild type strain, through their feeding of milk replacer. Neutrophil function will be monitored before and after day 0, the initiation of the trial. Colonization of mucosal surfaces of the nasal, vaginal, eye, ear, and rectum will be assessed throughout the trial. Colonization of the inner ear and upper and lower respiratory system will be assessed from postmortem samples. OBJECTIVE 2: Characterization and manipulation of virulence factors of mastitis pathogens for enhancing host defense. GA Assessment of S. aureus isolates misdiagnosed as coagulase-negative staphylococci will continue with emphasis on identifying specific characteristics that allow correct presumptive identification of such isolates based on specificity and sensitivity values. IA Work will center on metagenomic analysis of milk secretions from low SCC quarter and animals; and evaluate shifts in bacterial flora of the milk of normal quarters in response to new intramammary infections and challenge. LA Monitoring of Louisiana dairies for mycoplasma mastitis will continue. ME 1. Continue to test new isolates for heat resistance using standard methods of pasteurization, and investigate the phenotypic properties associated with resistance. 2. Evaluate prototheca in vitro sensitivity to available disinfectant and antibiotic compounds. MO We will be doing further evaluation of the potential differences in virulence between species of coagulase negative staphylococci use whole genome sequencing of selected strains. We will continue evaluating the risk of IMI post-calving in heifers. NY The Cornell team will continue to look at virulence characteristics of gram-negative organisms, particularly Klebsiella spp associated with clinical mastitis. TN We will continue with our research on pathogenesis of IMI caused by environmental pathogens, particularly S. uberis and E. coli, with ultimate goal to identify virulence factors with key role in the pathogenic process. VT We are investigating potential associations between carriage of antimicrobial resistance genes and response to antimicrobial therapy among different strain types of Staphylococcus aureus. VA Having identified differentially up-regulated proteins in pathogenic strains of S. aureus we will know genetically modify strains to knock out these genes and evaluate their role in pathogenesis. OBJECTIVE 3: Assessment and application of new technologies that advance mastitis control, milk quality and dairy food safety. GA A Direct Cell Counter (DCC) will be evaluated to determine its ability to correctly to enumerate somatic cells in goat milk in fresh versus frozen samples. IA 1) Continue to evaluate teat condition and health associated with new prototype lactating dips and compounds through our intense observational half udder design model, including histological evaluation of all tissue abnormalities. 2) Complete data collection, summarize, and publish results from the non-inferiority study examining the effects of 3 different dry cow antibiotics and their effects on prevention and cure of IMI across the dry period, calving, and early lactation, and clinical mastitis and production through 100 days in milk (in conjunction with MN and CA). 3) Continue to evaluate new compounds and dips for dry period mastitis prevention in dry cows and springing heifers. 4) We will begin a trial to prove efficacy of a new mastitis vaccine. KY A two-year study to examine the mastitis detection potential of multiple Precision Dairy Farming Technologies (MilkLine P4C individual quarter conductivity, IceRobotics IceQube lying behavior monitors, DVM Systems temperature boluses, SCR Rumination Tags) has started at the University of Kentucky. ME 1. Expand prototheca bulk tank evaluations to include a wider area: Collaborations with other investigators are being developed to recruit new dairies from outside the northeast region into the bulk tank filter project. 2. Continue validation of prototheca nested PCR: Split sampling between 2 or 3 laboratories for assay validation has been proposed with other investigators. MI Michigan will evaluate the application of RT-qPCR for Mycoplasma bovis detection in milk to herd mastitis programs, to be used as a method of surveillance of herd prevalence. MN 1. Continue working on the study: Development of an on-line curriculum in milk quality, mastitis control and treatment, and the analysis of milking equipment function on dairy farms. 2. Continue working on the study: Randomized Non-Inferiority Clinical Trial Evaluating Three Commercial Dry Cow Mastitis Preparations. MO In the coming year we will be evaluating a novel therapeutic regime for chronic Staphylococcus aureus IMI. NY The Cornell team will continue its work on monitoring and reducing food safety pathogens in bulk milk. UT Utah will continue surveillance for prevalence of infectious diseases of dairy cattle in Utah including Mycoplasma spp. Utah will conduct a statewide survey of Utah dairy farms for several bovine diseases using a repeated testing scheme for bulk tank milk samples. Utah will continue speciation using PCR of Mycoplasma spp. including M. gateae in milk samples from individual cows with subclinical and clinical mastitis and comparison to clinical severity and economic outcomes. VT We are investigating contagious mastitis transmission dynamics and control in automated milking systems. VA Data is being collected to evaluate the use of milk lactose concentration as an indicator of mastitis during naturally occurring mastitis. This data collection will be complete in 2012, followed by analysis and reporting. WA Develop and deliver an innovative outreach program to reduce the impact of mastitis on Western U.S. dairies.

NE-1028 ANNUAL REPORT  2011 ACCOMPLISHMENTS PEER-REVIEWED LITERATURE CONNECTICUT Ananda Baskaran, S., M. A. Roshni Amalaradjou, M. Procopio, Thomas Hoagland, G. Kazmer, S. M. Andrew and K Venkitanarayanan. 2012. Determining the efficacy of octenidine hydrochloride as a teat dip using excised teat model. J. Dairy Sci. (Submitted 12/28/2011) DELAWARE Nebzydoski, S. J., S. Pozzo, L. M. Nemec, M. K. Rankin, and T. F. Gressley. 2010. The effect of dexamethasone on clock gene mRNA levels in bovine neutrophils and lymphocytes. Vet. Immunol. Immunopathol. 138:183-192. GEORGIA Akers, R.M. and S. C. Nickerson. Mastitis and its impact on structure and function in the ruminant mammary gland. 2011. Journal of Mammary Gland Biology and Neoplasia. 16: Online First", 4 October 2011. Nickerson, S. C. Managing mastitis in heifers: An initial step in improving dairy herd health. 2011. In Milk Production / Book 1 (ISBN 979-953-307-116-8), InTech, Rijeka, Croatia. In press. KENTUCKY Russell, R.A. and J.M. Bewley. 2011. Producer assessment of dairy extension programming in Kentucky. J. Dairy Sci. 94: 2637-2647. MICHIGAN Erskine R.J., C. M. Corl, J. C. Gandy, and L. M. Sordillo. 2011. Effect of infection with bovine leukosis virus on lymphocyte proliferation and apoptosis in dairy cattle. Am J Vet Res. Aug;72(8):1059-64. Erskine RJ, P.C. Bartlett, K.M. Sabo, and L.M. Sordillo. Bovine Leukemia Virus Infection in Dairy Cattle: Effect on Serological Response to Immunization against J5 Escherichia coli Bacterin. Vet Med Intl. Volume 2011 (2011), Article ID 915747, 5 pages doi:10.4061/2011/915747 Mattmiller, S.A., C.M. Corl, J.C. Gandy, J.J. Loor, and L.M. Sordillo. 2011. Glucose transporter and hypoxia-associated gene expression in the mammary gland of transition dairy cattle. J. Dairy Sci. 94:2912-2922. Contreras, G.A. and L.M. Sordillo. 2011. Lipid mobilization and inflammatory responses during the transition period. Comp. Immunol. Microbiol. Infectious Dis. 34:281-289. Aitken, S.L., C.M. Corl, and L.M. Sordillo. 2011. Pro-inflammatory and pro-apoptotic gene expression of TNF-a stimulated bovine mammary endothelial cells. Vet. Immunol. Immunopathol. 140:282-290. Hill, T.M., M.J. VandeHaar, L.M. Sordillo, D.R. Catherman, H.G. Bateman, and R.L. Schlotterbeck. 2011. Fatty acid intake alters growth and immunity in milk-fed calves. J. Dairy Sci. 94:3936-3948. Aitken, S.L., C.M. Corl, and L.M. Sordillo. 2011. Immunopathology of Mastitis: Insights into disease recognition and resolution. J. Mammary Gland Neoplasia. Epub ahead of print: DOI: 10.1007/s10911-011-9230-4 Sordillo, L.M. 2011. New concepts in the causes and control of mastitis. J. Mammary Gland Neoplasia. Epub ahead of print: DOI: 10.1007/s10911-011-9239-8 Sordillo, L.M. and S. L. Aitken 2011. Mammary Resistance Mechanisms - Endogenous. In: Fuquay, J.W., Fox, P.F. and McSweeney, P.L.H. (eds), Encyclopedia of Dairy Sciences, Second Edition, vol. 3, pp. 386-391. San Diego: Academic Press. Contreras, G.A. and L.M. Sordillo. 2011. Lipid mobilization and inflammatory responses during the transition period. Comp. Immunol. Microbiol. Infectious Dis. 34:281-289. Contreras, G.A. and J.M. Rodriguez. 2011. Mastitis: Comparative Etiology and Epidemiology. J. Mammary Gland Neoplasia. DOI: 10.1007/s10911-011-9234-0 Contreras, G.A. 2011Mycoplasma bovis in dairy cattle. Mycoplasma bovis en el ganado lechero. Albeitar 26:28-30. MINNESOTA Haran, K.P., S.M. Godden, D. Boxrud, J.B. Bender, S. Sreevatsan. Isolation and Characterization of Methicillin Resistant Staphylococcus aureus from Bulk Tank Milk in Minnesota Dairy Farms. Submitted to J. Submitted: Clinical Microbiology. Lago, A., S.M. Godden, R. Bey, P.L. Ruegg, K. Leslie. The selective treatment of clinical mastitis based on on-farm culture results I: Effects on antibiotic use, milk withholding time and short-term clinical and bacteriological outcomes. Accepted J. Dairy Sci. Lago, A., S.M. Godden, R. Bey, P.L. Ruegg, K. Leslie. The selective treatment of clinical mastitis based on on-farm culture results II: Effects on lactation performance including, clinical mastitis recurrence, somatic cell count, milk production and cow survival. Accepted J. Dairy Sci. Lievaart, J.J., J.K. Reneau, W.D.J. Kremer and H.W. Barkema. 2011. Short communication: Influence of sampling interval on the accuracy of predicting bulk milk somatic cell count. JDSci. 94:804-807. MISSOURI Zadoks RN, Middleton JR, McDougall S, Katholm J, Schukken YH. Molecular epidemiology of mastitis pathogens of dairy cattle and comparative relevance to humans. J Mammary Gland Biol Neoplasia. Epub ahead of print Oct 2011. NEW YORK Schukken YH, Bennett GJ, Zurakowski MJ, Sharkey HL, Rauch BJ, Thomas MJ, Ceglowski B, Saltman RL, Belomestnykh N, Zadoks RN. Randomized clinical trial to evaluate the efficacy of a 5-day ceftiofur hydrochloride intramammary treatment on nonsevere gram-negative clinical mastitis. J Dairy Sci. 2011 Dec;94(12):6203-15. 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Vet Microbiol. 2011 May 12;150(1-2):173-9. Epub 2011 Jan 28. PubMed PMID: 21333468. Zadoks RN, Griffiths HM, Munoz MA, Ahlstrom C, Bennett GJ, Thomas E, Schukken YH. Sources of Klebsiella and Raoultella species on dairy farms: be careful where you walk. J Dairy Sci. 2011 Feb;94(2):1045-51. PubMed PMID: 21257074. Almeida RA, Dogan B, Klaessing S, Schukken YH, Oliver SP. Intracellular fate of strains of Escherichia coli isolated from dairy cows with acute or chronic mastitis. Vet Res Commun. 2011 Feb;35(2):89-101. Epub 2011 Jan 5. PubMed PMID: 21207146. Dohoo IR, Smith J, Andersen S, Kelton DF, Godden S; Mastitis Research Workers' Conference. Diagnosing intramammary infections: evaluation of definitions based on a single milk sample. J Dairy Sci. 2011 Jan;94(1):250-61. PubMed PMID: 21183035. Schukken YH, Bar D, Hertl J, Gröhn YT. Correlated time to event data: Modeling repeated clinical mastitis data from dairy cattle in New York State. Prev Vet Med. 2010 Dec 1;97(3-4):150-6. Epub 2010 Oct 28. PubMed PMID: 21035216. Andersen S, Dohoo IR, Olde Riekerink R, Stryhn H; Mastitis Research Workers' Conference. Diagnosing intramammary infections: evaluating expert opinions on the definition of intramammary infection using conjoint analysis. J Dairy Sci. 2010 Jul;93(7):2966-75. PubMed PMID: 20630213. Latorre AA, Van Kessel JS, Karns JS, Zurakowski MJ, Pradhan AK, Boor KJ, Jayarao BM, Houser BA, Daugherty CS, Schukken YH. Biofilm in milking equipment on a dairy farm as a potential source of bulk tank milk contamination with Listeria monocytogenes. J Dairy Sci. 2010 Jun;93(6):2792-802. PubMed PMID: 20494189. Zanella R, Settles ML, McKay SD, Schnabel R, Taylor J, Whitlock RH, Schukken Y, Van Kessel JS, Smith JM, Neibergs HL. Identification of loci associated with tolerance to Johne's disease in Holstein cattle. Anim Genet. 2011 Feb;42(1):28-38. doi: 10.1111/j.1365-2052.2010.02076.x. PubMed PMID: 20477805. Scaccabarozzi L, Locatelli C, Pisoni G, Manarolla G, Casula A, Bronzo V, Moroni P. Short communication: Epidemiology and genotyping of Candida rugosa strains responsible for persistent intramammary infections in dairy cows. J Dairy Sci. 2011 Sep;94(9):4574-7. PubMed PMID: 21854930. Zonca A, Gallo M, Locatelli C, Carli S, Moroni P, Villa R, Cagnardi P. Cefquinome sulfate behavior after intramammary administration in healthy and infected cows. J Dairy Sci. 2011 Jul;94(7):3455-61. PubMed PMID: 21700031. Genini S, Badaoui B, Sclep G, Bishop SC, Waddington D, Pinard van der Laan MH, Klopp C, Cabau C, Seyfert HM, Petzl W, Jensen K, Glass EJ, de Greeff A, Smith HE, Smits MA, Olsaker I, Boman GM, Pisoni G, Moroni P, Castiglioni B, Cremonesi P, Del Corvo M, Foulon E, Foucras G, Rupp R, Giuffra E. Strengthening insights into host responses to mastitis infection in ruminants by combining heterogeneous microarray data sources. BMC Genomics. 2011 May 11;12(1):225. PubMed PMID: 21569310; PubMed Central PMCID: PMC3118214. Cuccuru C, Meloni M, Sala E, Scaccabarozzi L, Locatelli C, Moroni P, Bronzo V. Effects of intramammary infections on somatic cell score and milk yield in Sarda sheep. N Z Vet J. 2011 May;59(3):128-31. PubMed PMID: 21541886. Ricchi M, Goretti M, Branda E, Cammi G, Garbarino CA, Turchetti B, Moroni P, Arrigoni N, Buzzini P. Molecular characterization of Prototheca strains isolated from Italian dairy herds. J Dairy Sci. 2010 Oct;93(10):4625-31. PubMed PMID: 20854996. Cagnardi P, Villa R, Gallo M, Locatelli C, Carli S, Moroni P, Zonca A. Cefoperazone sodium preparation behavior after intramammary administration in healthy and infected cows. J Dairy Sci. 2010 Sep;93(9):4105-10. PubMed PMID: 20723685. Locatelli C, Scaccabarozzi L, Pisoni G, Moroni P. CTX-M1 ESBL-producing Klebsiella pneumoniae subsp. pneumoniae isolated from cases of bovine mastitis. J Clin Microbiol. 2010 Oct;48(10):3822-3. Epub 2010 Aug 18. PubMed PMID: 20720020; PubMed Central PMCID: PMC2953115. Pisoni G, Moroni P, Genini S, Stella A, Boettcher PJ, Cremonesi P, Scaccabarozzi L, Giuffra E, Castiglioni B. Differentially expressed genes associated with Staphylococcus aureus mastitis in dairy goats. Vet Immunol Immunopathol. 2010 Jun 15;135(3-4):208-17. Epub 2009 Dec 21. PubMed PMID: 20060596. OHIO Zimov, J.L., N.A. Botheras, W.P. Weiss, and J.S. Hogan. 2011. Associations among behavioral and acute physiological responses to lipopolysaccharide induced clinical mastitis. Amer. J. Vet. Res. 72:620-627. Petersson-Wolfe, C.S., S. Masiello, and J.S. Hogan. 2011. The ability of mastitis-causing pathogens to survive as ensiling period. J. Dairy Sci. 94:5027-5032. TENNESSEE Almeida, R. A., D. A. Luther, D. Patel, and S. P. Oliver. 2011. Predicted antigenic regions of Streptococcus uberis adhesion molecule (SUAM) are involved in adherence to and internalization into mammary epithelial cells. Vet. Microbiol. 148:323-28. Almeida, R.A., B. Dogan, S. Klaessing, Y. H. Schukken and S. P. Oliver. 2011. Intracellular fate of strains of Escherichia coli isolated from dairy cows with acute or chronic mastitis. Vet. Res. Comm. 35:89101. Chen, Xueyan, Oudessa Kerro Dego, Troy E. Fuller, Raul A. Almeida, Douglas A. Luther, and Stephen P. Oliver. 2011. Deletion of sua gene reduces the ability of Streptococcus uberis to adhere to and internalize into bovine mammary epithelial cells. Vet. Microbiol. 147:426-434. Kerro Dego, O., R. A. Almeida, and S.P. Oliver. 2011. Presence of ISS1-like insertion sequence in wild type Streptococcus uberis strains isolated from cases of bovine mastitis. Vet. Microbiol. 151: 315320 Kerro Dego, O., S. P. Oliver, and R. A. Almeida. 2011. Hostpathogen gene expression profiles during infection of primary bovine mammary epithelial cells with Escherichia coli strains associated with acute or persistent bovine mastitis. Vet. Microbiol. doi:10.1016/j.vetmic.2011.08.016. Kerro-Dego O, M. E. Prado, X. Chen, D. A. Luther D.A., R. A. Almeida RA, and S. P. Oliver. 2011. pGh9:ISS1 transpositional mutations in Streptococcus uberis UT888 causes reduced bacterial adherence to and internalization into bovine mammary epithelial cells. Vet Microbiol. 151: 379-85. Epub 2011 Apr 12. Oliver, S. P., S. E. Murinda, and B. M. Jayarao. 2011. Impact of antibiotic use in adult dairy cows on antimicrobial resistance of veterinary and human pathogens: A comprehensive review. Foodborne Pathogens & Disease 8(3): 337-355. Oliver, S. P., and S. E. Murinda. 2011. Milk and raw milk consumption as a vector for human disease. p. 99-118 In: Zoonotic Pathogens in the Food Chain, Denis Krause and Steve Hendrick Eds., CAB International, United Kingdom. Prado ME, Almeida R.A., Ozen C., Luther D.A., Lewis M.J., Headrick S.J., Oliver S.P. 2011. Vaccination of dairy cows with recombinant Streptococcus uberis adhesion molecule induces antibodies that reduce adherence to and internalization of S. uberis into bovine mammary epithelial cells. Vet Immunol Immunopathol. 141: 201-8. Pighetti GM and AA Elliott. 2011. Gene polymorphisms: The keys for marker assisted selection and unraveling core regulatory pathways for mastitis resistance. J Mammary Gland Biol Neoplasia. 16:421-432. PMID 22056270. Pighetti GM, C Kojima, L. Wojakiewicz, and M Rambeaud. 2011. The bovine CXCR1 gene is highly polymorphic. Vet Immunol Immunopathol. http://dx.doi.org/10.1016/j.vetimm.2011.09.012 PMID 22056270. Schrick, F. N., S. P. Oliver, and J. L. Edwards. 2011. Mastitis during early lactation alters reproductive performance of dairy cows. Alteración del rendimiento reproductive en la lactación temprana. Invited Review. Albéitar 145:14-17. UTAH Wilson DJ, Justice-Allen A, Goodell G, Baldwin TJ, Skirpstunas RT, Cavender KB: Risk of Mycoplasma bovis transmission from contaminated sand bedding to naive dairy calves. J Dy Sci 94:3:1318-1324, 2011. Justice-Allen A, Trujillo J, Goodell G, Wilson D: Detection of multiple Mycoplasma species in bulk tank milk samples using real-time PCR and conventional culture and comparison of test sensitivities. J Dy Sci 94:3411-3419, 2011. Maunsell FP, Woolums AR, Francoz D, Rosenbusch RF, Step DL, Wilson DJ, Janzen ED: Mycoplasma bovis infections in cattle. J Vet Intern Med 25:4:772-783, 2011. Wilson DJ, Terra RL, Muller FJ, Sears PM, Anderson KL: Milk quality and udder health on organic dairies. Am Assoc Bov Pract Fact Sheet: 1-3, May, 2011. VERMONT Kandasamy, S., B.B. Green, A.L. Benjamin, and D.E. Kerr. 2011. Between-cow variation in dermal fibroblast response to lipopolysaccharide reflected in resolution of inflammation during Escherichia coli mastitis. Journal of Dairy Science. In press. Green, B.B., S. Kandasamy, T.H. Elsasser, and D.E. Kerr. 2011. The use of dermal fibroblasts as a predictive tool of the toll-like receptor 4 response pathway and its development in Holstein heifers. Journal of Dairy Science. 94:5502-5514. Becker, S.C., J. Foster-Frey, A. Powell, D.E. Kerr, and D. M. Donovan. 2011. Lysostaphin: molecular changes that preserve staphylolytic activity. In "Science and Technology against Microbial Pathogens. Research, Development and Evaluation" A. Mendez-Vilas ed. World Scientific Publishing Co. pp18-22. VIRGINIA Petersson-Wolfe C. S., S. Masiello and J. S. Hogan. 2011. The ability of common mastitis-causing pathogens to survive an ensiling period. J. Dairy Sci. 94(10):5027-32. Petersson K. H., L. A. Connor, C. S. Petersson-Wolfe and K. A. Rego. 2011. Evaluation of confirmatory stains used for direct microscopic somatic cell counting of sheep milk. J. Dairy Sci. 94(4):1908-12. Maxymiv N.G., M. Bharathan, I.K. Mullarky. Bovine mammary dendritic cells: A heterogeneous population, distinct from macrophages and similar in phenotype to afferent lymph veiled cells. Comp Immunol Microbiol Infect Dis. 2011 Oct 20. PMID: 22019401. Bharathan, M., I.K. Mullarky. Targeting mucosal immunity in the battle to develop a mastitis vaccine. Journal: Journal of Mammary Gland Biology and Neoplasia. Dec;16(4):409-19. Epub 2011 Oct 4. PMID: 21968537. Sordillo LM, I.K. Mullarky. Cutting edge technologies in animalomics. Vet Immunol Immunopathol. 2010 Dec 15;138(4):239-40. Epub 2010 Oct 14. PMID: 21036405 (1.963) WASHINGTON Madhusoodanan J., K.S. Seo, B. Remortel, J.Y. Park, S.Y. Hwang, L.K. Fox LK, Y.H. Park, C.F. Deobald, D. Wang S. Liu , S.C. Daugherty, A.L. Gill, G.A.Bohach,and S.R. Gill, 2011. An enterotoxin-bearing pathogenicity island in Staphlococcus epidermidis. J. bacteriology. 193:1854-62. Punyaportwithaya, V., L.K. Fox, D. D. Hancock, J.M. Gay, J.R. Wenz, J. R. Alldredge. 2011. Incidence and transmission of Mycoplasma bovis mastitis in Holstein dairy cows in a hospital pen: A case study. Prev. Vet. Med. 98:74-78. Hunt, K.M., Hunt, K.M., J. A. Foster, L.J. Forney, U.M.E. Shute, D.L. Beck, Z. Abdo, L,K. Fox, J.E. Williams, M.K. McGuire, M.A. McGuire. 2011. Characterization of the Diversity and Temporal Stability of Bacterial Communities inHuman Milk. PLoS ONE 6(6): e21313. doi:10.1371/journal.pone.0021313 Wenz, J.R., L.K. Fox, F.J. Muller, M. Rinaldi, R. Zeng, and D. D. Bannerman. 2010. Factors associated with concentrations of select cytokine and acute phase proteins in dairy cows with naturally occurring clinical mastitis. J. Dairy Sci. 93:2458-2470. Adhikari, B., T.E. Besser, J.M. Gay, L.K. Fox, D. D. Hancock, and M. A. Davis. 2010. Multilocus variable-number-tandem-repeat analysis and plasmid profiling to study the occurrence of blacmy-2 within a pulsed field gel electrophoresis defined cade of Salmonella enterica serovar Typhimurium. Appl. Envir. Microbiol. 76:69-74. Park, J.Y., L.K. Fox, K.S. Seo, M. A. McGuire, Y.H. Park, F.R. Rurangirwa, W.M. Sischo, G. A. Bohach. 2010. Detection of classical and newly described staphylococcal superantigen genes in coagulase-negative staphylococci isolated from bovine intramammary infections. Vet. Micro. 147:149-154. Park, J.Y., L.K. Fox, K.S. Seo, M. A. McGuire, Y.H. Park, F.R. Rurangirwa, W.M. Sischo, G. A. Bohach. 2010. Comparison of phenotypic and genotypic methods for the species identification of coagulase-negative staphyloccal isolates from bovine intramammary infections. Vet. Micro. 147:142-148. Boonyayatra, S., LK. Fox, T.E. Besser, A. Sawant, and J.M. Gay. 2010. Effects of storage methods on the recovery of Mycoplasma species from milk samples. Vet. Micro. 144:210-213. Punyapornwithaya, V., L.K. Fox, J.M. Gay, D.D. Hancock, and J.R. Alldredge. 2010. Association between an outbreak strain causing mycoplasma bovis mastitis and its asymptomatic carriage in the herd: A case study from Idaho, USA. Prev. Vet. Med. 93:66-70. ABSTRACTS DELAWARE Nemec L.M., C. Wu, S. Cordova, K. Davison, and T. F. Gressley. 2010. The effect of thymol on intracellular and extracellular reactive oxygen species production by bovine neutrophils. J. Dairy Sci. 93(E-Suppl. 1):313. GEORGIA Jackson, K.A.*, S.C. Nickerson, F.M. Kautz, and D.J. Hurley. 2011. Experimentally induced Streptococcus uberis mastitis in dairy heifers: A challenge model incident to a vaccine trial. Abstract. ADSA Annual Meeting. New Orleans, LA. Journal of Dairy Science 94, E-supplement 1: 701. Eubanks, V.J.*, N.E. Forsberg, Y.Q. Wang, K.P. Zanzalari, J.D. Chapman, D.J. Hurley, F.M. Kautz, L.O. Ely, and S.C. Nickerson. 2011. Effects of OmniGen-AF on enhancing immunity in dairy heifers vaccinated with a Staphylococcus aureus bacterin. Abstract. ADSA Annual Meeting. New Orleans, LA. Journal of Dairy Science 94, E-supplement 1: 702-703. IDAHO Rezamand. P., K. M. Hunt, R. D. Schramm, and M. A. McGuire. 2010. Bovine hepatic and adipose retinol binding protein gene expression. J. Dairy Sci. Vol. 93 (E-Suppl. 1):313 (Abstr.). KENTUCKY Bewley, J.M. 2011. Extension programming in Kentucky to address somatic cell count challenges and opportunities. Abstract 170. American Dairy Science Association Annual Meeting. New Orleans, Louisiana. Sterrett, A.E. and J.M. Bewley. 2011. Characterization of management practices utilized by low somatic cell count Kentucky dairy herds. Abstract M160. American Dairy Science Association Annual Meeting. New Orleans, Louisiana. Black, R.A., J.L. Taraba, G.B. Day, F. A. Damasceno, M. C. Newman. K. A. Akers, and J.M. Bewley. 2011. Relationships among temperature, moisture, bacterial counts, and animal hygiene in compost bedded pack barns. Abstract 234. American Dairy Science Association Annual Meeting. New Orleans, Louisiana MINNESOTA Husfeldt, A.W., M.I. Endres, K. A. Janni, J.A. Salfer, J.K. Reneau. Chemical and bacteriological characteristics of digested, composted, and separated raw manure solids prior to use as freestall bedding. Annu mtg ADSA. July 10-14, 2011. New Orleans, LA. Husfeldt, A.W., M.I. Endres, J.A. Salfer, and J.K. Reneau. Mortality and herd turnover rates in dairy herds utilizing recycled manure solids for bedding freestalls. Annu mtg ADSA. July 10-14, 2011. New Orleans, LA. Leuer, R.F. and J. K. Reneau. Effect of season on four categories of fresh and current new mastitis infections in Minnesota. Annu mtg ADSA. July 10-14, 2011. New Orleans, LA. Leuer, R.F. and J. K. Reneau. Somatic Cell Count and Management Benchmarks in Minnesota Dairy Herds. Annu mtg ADSA. July 10-14, 2011. New Orleans, LA. Haran, K.P., S. M. Godden, J. B. Bender, and S. Sreevatsan. Prevalence of Methicillin Resistant Staphylococcus aureus in Bulk Tank Milk of Minnesota Dairy Farms  Significance and Risk Factor Analysis. Annu mtg MN Dairy Health Conference. May 17-19, 2011. Bloomington, MN MISSOURI Middleton JR, Perry J, Zadoks RN, Dufour S, Paradis ME, Scholl DT. 2011. A new look at the importance of coagulase negative staphylococci in bovine mastitis. ACVIM Forum. Denver, Colorado. June 15-18, 2011. Ringen DR, Middleton JR, Suntrup D, Markovich T. 2011. Epidemiology of mastitis pathogens in heifers on a grazing dairy. Abstract FA-2. ACVIM Forum. Denver, Colorado June. 15-18, 2011. Dank A, Middleton JR. 2011. Efficacy of two teat skin disinfection techniques. Veterinary Research Scholars Symposium. Orlanda, Florida. August 2011. Paradis, M.-È., Haine, D., Messier, S., Middleton, J., Perry, J., Ramirez, A.I. and Scholl, D.T. 2011. Identification of coagulase-negative Staphylococcus species by gas chromatography. International Conference on Udder Health and Communication. Utrecht, Netherlands. Oct 25-57, 2011. TENNESSEE Almeida , R. A., X. Chen, M. E. Prado, and S. P. Oliver. 2011. Role of Streptococcus uberis adhesion molecule in adherence to and internalization of Streptococcus uberis into bovine mammary epithelial cells. AABP Third International Symposium on Mastitis and Milk Quality. St. Louis, Missouri USA. Kerro Dego, O., R. A. Almeida, and S. P. Oliver. 2011. Identification of virulence-associated genes and their expression patterns in strains of Escherichia coli associated with bovine mastitis. AABP Third International Symposium on Mastitis and Milk Quality. St. Louis, Missouri USA Kerro Dego, O., R. A. Almeida, and S.P. Oliver. 2011 Gene expression of bovine mammary epithelial cells infected with Escherichia coli associated with acute or persistent bovine mastitis. AABP Third International Symposium on Mastitis and Milk Quality. St. Louis, Missouri USA. Kerro Dego, O., S. P. Oliver and R. A. Almeida. 2011. The Expression Patterns of Virulence Associated Genes of E. coli Strains Associated with Acute and Chronic Bovine Intramammary Infections after Co-culture with Mammary Epithelial Cells. Paradis, Marie-Ève, Denis Haine, Stephen P. Oliver, Barbara Gillespie, Sylvain Messier, Josaphat Comeau, and Daniel T. Scholl. 2011. Estimation of the diagnostic accuracy of a multiplex real-time pcr assay and bacteriological culture for four bovine intramammary pathogens. Proc. Canadian Assoc. Vet. Epidemiol. Prev. Med. In press. Prado, Maria E., Glenis Moore, Oudessa Kerro-Dego, Raul Almeida, and Stephen Oliver. 2011. Streptococcus uberis mastitis isolates vary in their ability to form biofilms. Proc. NMC/AABP Intl. Meeting on Mastitis and Milk Quality. Submitted. Siebert LJ, J Lippolis, and GM Pighetti. 2011. Expression of CXCR1 and CXCR2 in bovine mammary tissue. In Proc. Conference of Research Workers in Animal Diseases. Warren, J O. Kerro Dego, R.A. Almeida, S.P. Oliver, and G.M. Pighetti. 2011. Identification of Streptococcus uberis mutants susceptible to neutrophil killing. 92th CRWAD Annual Meeting, Chicago, IL UTAH Wilson DJ, Justice-Allen A, Baldwin TJ, Skirpstunas RT, Cavender KB, Goodell G: An experiment in transmission of Mycoplasma bovis in sand bedding to naive dairy calves. J Dy Sci Vol. 93, E-Suppl. 1:253, 2010. (Published since last NE-1028 report.) Wilson DJ, Justice-Allen A, Trujillo JD, Goodell G: Multiple Mycoplasma spp. detected in bulk tank milk samples using real-time PCR and conventional culture, and agreement between test methods. J Dy Sci Vol. 94, E-Suppl. 1:701, 2011. VERMONT Green B.B., S. Kandasamy, and D.E. Kerr. 2010. Use of dermal fibroblasts to evaluate developmental, and between-animal variation in innate immune response. 91st Annual Meeting of the Conference of Research Workers in Animal Diseases. Chicago, IL. Kandasamy, S., B. B., Green, and D.E. Kerr. 2010. Response of cultured fibroblasts to LPS reflects in vivo response to E. coli mastitis. 91st Annual Meeting of the Conference of Research Workers in Animal Diseases. Chicago, IL. Kerr, D.E. and S. Kandasamy. 2010. Genomic analysis of LPS-challenged bovine dermal fibroblasts to evaluate inter-animal variation in innate immunity. 2nd International Symposium on Animal Health Genomics for Animal Health. Paris, France. VIRGINIA Yeiser, E. E., K. E. Leslie and C. S. Petersson-Wolfe. 2011. The effects of flunixin meglumine treatment on activity measures and feed intake during experimentally induced Escherichia coli mastitis. National Mastitis Council 3rd International Symposium on Mastitis and Milk Quality, St. Louis, MO. Pp. 32-36. Fitzpatrick, C. E., K. E. Leslie, T. F. Duffield, D. F. Kelton, T. J. DeVries and C. S. Petersson-Wolfe. 2011. Objective assessment of pan in dairy cattle with clinical mastitis. National Mastitis Council 3rd International Symposium on Mastitis and Milk Quality, St. Louis, MO. Pp. 27-31. Ortiz Marty, R.J., W. Wark, I. Mullarky. 2011. Staphylococcus aureus virulence cluster agr- regulated hemolysins dictate bovine polymorphonuclear leukocyte inflammatory signaling. Proceedings of the 92nd Annual Conference of Research Workers in Animal Disease. Manickam, M., I.K. Mullarky. 2011 Detecting differential proteome expression between pathogenic and commensal Staphylococcus aureus strains using SILAC. Proceedings of the 92nd Annual Conference of Research Workers in Animal Disease. WASHINGTON Alabdullah, H., S.S. Schneider, and L. Fox. 2011. Effect of dexamethasone administration on shedding of Mycoplasma bovis in calves. Third International Symposium on Mastitis and Milk Quality, St. Louis, MO, Septmeber 22-24. Boonyayatra, S., L.K. Fox, T.E. Besser, A. Sawant, J. M. Gay, and Z. Raviv. 2011. Identification of Mycoplasma sp. using novel reatl-time PCR assays. Third International Symposium on Mastitis and Milk Quality, St. Louis, MO, Septmeber 22-24. Scheneider, C.S., L,K. Fox, and A. N. Hristov. 2011. The effects of lauric acid on mastitis pathogens. Proceedings of the NMC 50th Annual Meeting, Arlington, VA. pages 199-200. CONFERENCE PROCEEDINGS GEORGIA Jackson, K.A., S.C. Nickerson, F.M. Kautz, and D.J. Hurley. 2011. Experimentally induced Streptococcus uberis mastitis in dairy heifers. Page 164 in 3rd International Symposium on Mastitis and Milk Quality and American Association of Bovine Practitioners 44th Annual Conference, September 22-24, 2011, St. Louis, MO. Nickerson, S. C. 2011. Development of mastitis. in Current Concepts of Bovine Mastitis. National Mastitis Council, Inc. Madison, WI. In Press. Nickerson, S.C. and R.M. Akers. 2011. Mammary Gland | Anatomy. In: Fuquay, J.W., P. F. Fox, and P.L.H. McSweeney (eds.), Encyclopedia of Dairy Sciences, Second Edition, vol. 3, pp. 328337. San Diego: Academic Press. Nickerson, S.C. 2011. Mammary Resistance Mechanisms | Anatomical. In: Fuquay, J.W., P. F. Fox, and P.L.H. McSweeney (eds.), Encyclopedia of Dairy Sciences, Second Edition, vol. 3, pp. 381385. San Diego: Academic Press. Nickerson, S.C. 2011. Mastitis Pathogens | Contagious Pathogens. In: Fuquay, J.W., P. F. Fox, and P.L.H. McSweeney (eds.), Encyclopedia of Dairy Sciences, Second Edition, vol. 3, pp. 408414. San Diego: Academic Press. Nickerson, S.C. and W.E. Owens. (2011) Mastitis Therapy and Control | Management Control Options. In: Fuquay JW, Fox PF and McSweeney PLH (eds.), Encyclopedia of Dairy Sciences, Second Edition, vol. 3, pp. 429434. San Diego: Academic Press. Owens, W. E. and S. C. Nickerson. 2011. Mastitis Therapy and Control | Medical Therapy Options. In: Fuquay JW, Fox PF and McSweeney PLH (eds.), Encyclopedia of Dairy Sciences, Second Edition, vol. 3, pp. 435439. San Diego: Academic Press. IDAHO Rezamand, P., M. A. McGuire, and S. M. Andrew. 2009. Interaction among energy status, retinol-binding protein and intra-mammary infections in periparturient dairy cows. In: Proceedings of the 44th Annual Pacific Northwest Animal Nutrition Conference. pp. 71-83. IOWA Lopez, M., K. Knutson, L. Timms, M. Henderson, and T. Hemling. 2011. Evaluation of teat conditioning using liquid or powder dips in winter. NMC Annual Proceedings: 181. MICHIGAN RJ Erskine Mastitis Therapy: Quality Milk Short Course for Dairy Practitioners, - St Peters, MN (Univ of Minnesota) July, 2011. RJ Erskine - Milk Quality and Mastitis Control: Annual National Conference of the American Dairy Goat Association, Grand Rapids, MI, October, 2011. RJ Erskine - Mastitis Therapy and Mastitis Vaccination: Quality Milk Short Course of the Dairy Production Medicine Program for Dairy Practitioners, State College, PA (Pennylvania State University) October, 2011. RJ Erskine - Mastitis Pharmacology: NOSAI Japan- Veterinary Shortcourse, East Lansing, MI, Oct, 2011. RJ Erskine - Antimicrobial Resistance in the Dairy Industry: Presented to the Antimicrobial Stewardship in Canadian Agriculture and Veterinary Medicine Conference, Toronto, ONT, Nov, 2011. LM Sordillo - Presented invited seminar entitled Nutrient metabolism and oxidative stress in periparturient dairy cows: impact on disease susceptibility at the Annual Conference of the Japan Veterinary Medical Association, Gifu, Japan. February 2011. LM Sordillo - Presented invited seminar entitled Oxidative stress and mastitis susceptibility: The immunology Link at the Pacific Northwest Animal Nutrition and Management Conference, Portland, Oregon. October 2011. LM Sordillo - Presented invited seminar entitled Inflammation in the transition cow: Is it harmful or helpful at the Pacific Northwest Animal Nutrition and Management Conference, Portland, Oregon. October 2011. MINNESOTA Lago A., Godden S.M., Bey R., Ruegg P., Leslie K. (2011). Efficacy of the Selective Treatment of Clinical Mastitis Based on On-Farm Culture Results. In: Proceedings of the International Conference on Udder Health and Communication, Utrecht, The Netherlands, pp 277-284. Lago A., Godden S.M., Bey R., Leslie K., Ruegg P. (2011). Efficacy of Two Fresh Cow Subclinical Mastitis Treatment Programs. In: In: Proceedings of the International Conference on Udder Health and Communication, Utrecht, The Netherlands, pp 293. MISSOURI Ringen DR, Suntrup D, Middleton JR. 2011. Molecular characterization of isolates from pre-partum secretions, teat skin swabs and post-partum milk samples from replacement heifers on a grazing dairy. Proc. 50th Annual Meeting of the Natl. Mast. Counc. Arlington, VA. Jan 23-26, 2010. 127-128. Ringen DR, Middleton JR, Suntrup D, Markovich T. 2011. Epidemiology of mastitis pathogens in heifers on a grazing dairy. Proc. 3rd Intl. Mastitis and Milk Quality Symposium. St. Louis, MO. Sept 22-24, 2011. Chamberlin W, Middleton JR, Spain JN, Johnson GC, Ellersieck MR. 2011. Association Between Subclinical Hypocalcemia, Milk Somatic Cell Count, and Clinical Mastitis. Proc. 3rd Intl. Mastitis and Milk Quality Symposium. St. Louis, MO. Sept 22-24, 2011. TENNESSEE Hogeveen, H., S. Pyorala, Karin Persson-Waller, J. S. Hogan. T. J. G. M. Lam, S. P. Oliver, Y. H. Schukken, H. W. Barkema, and J. E. Hillerton. 2011. Current status and future challenges in mastitis research. In: Proc. Natl. Mastitis Council, pp. 36-48. Paradis, M. E., S. P. Oliver, B. Gillepsie, S. Messier, J. Comeau, and D.T. Scholl. 2011. Estimation of the diagnostic accuracy of a multiplex real-time PCR assay and bacteriological culture for four bovine intramammary pathogens. In: Proc. Natl. Mastitis Council, pp.117-118. UTAH Wilson DJ, Justice-Allen A, Goodell G, Baldwin TJ, Skirpstunas RT, Cavender KB: Is Mycoplasma bovis in bedding sand transmissible to naive dairy calves? Proc 43rd Ann Conv Am Assoc Bov Pract: 201, 2010. (Published since last NE-1028 report.) Wilson DJ, Justice-Allen A, Goodell G, Baldwin TJ, Skirpstunas RT, Cavender KB: Experimental exposure of naive dairy calves to Mycoplasma bovis in naturally contaminated bedding sand  risk of transmission. Proc 53rd Ann Conf Am Assoc Vet Lab Diag: 131, 2010. (Published since last NE-1028 report.) Wilson DJ, Justice-Allen AE, Trujillo JD, Goodell G: Detection of Mycoplasma spp. in bulk tank milk samples using conventional culture and real-time PCR. Proc Natl Mast Council - Am Assoc Bov Pract 3rd International Symposium on Mastitis and Milk Quality, 125-129, 2011. VERMONT Kerr, D.E., B.B. Green, and S. Kandasamy. 2011. Variation in cow response to mastitis predicted by laboratory testing of skin cells. Proceedings of the 50th Annual Meeting of the National Mastitis Council. Arlington, VA. pages 64-69. VIRGINIA Yeiser, E. E., K. E. Leslie and C. S. Petersson-Wolfe. 2011. The effects of flunixin meglumine treatment on activity measures and feed intake during experimentally induced Escherichia coli mastitis. National Mastitis Council Annual Meeting 2011, Arlington, VA. Pg 133-134. Fitzpatrick, C. E., K. E. Leslie, T. F. Duffield, D. F. Kelton, T. J. DeVries and C. S. Petersson-Wolfe. 2011. Objective assessment of pan in dairy cattle with clinical mastitis. National Mastitis Council Annual Meeting 2011, Arlington, VA. Pg 163-164. WEB PRESENTATIONS POSTER PRESENTATIONS GEORGIA Eubanks V*., N. Forsberg, Y. Wang, K. Zanzalari, J. Chapman, L. Ely, and S.C. Nickerson. 2011. Immunostimulating effects of OmniGen-AF on vaccinating dairy heifers with a Staphylococcus aureus bacterin: Preliminary report. Exhibit (Poster). Georgia Farm Bureau Commodity Conference Tour, Athens, GA. M. Upchurch*, M., F. Kautz, and S.C. Nickerson. Local leukocyte response of the heifer mammary gland to mastitis-causing bacterial pathogens. (Poster) CAES Young Scholars Program. July 6, 2011. IDAHO Rezamand. P., K. M. Hunt, R. D. Schramm, and M. A. McGuire. 2010. Bovine hepatic and adipose retinol binding protein gene expression. ADSA- ASAS joint annual meeting, Denver CO IOWA Lopez, M., K. Knutson, L. Timms, M. Henderson, and T. Hemling. 2011. Evaluation of teat conditioning using liquid or powder dips in winter. NMC Annual Meeting. Arlington, VA. MAINE Farrington-Thomason, T and Lichtenwalner, AB. Pasteurization: Effective Control of Prototheca spp. Human Health Risks? University of Maine Graduate Student poster competition, May 2011. Prototheca mastitis in Maine: prevalence and pasteurization studies of a food-borne pathogen. Anne Lichtenwalner, Nirajan Adhikari, Tanya Farrington-Thomason. Annual Infectious Diseases Conference: Maine CDC, Nov. 2011. MINNESOTA Haran, K.P., S. M. Godden, D. Boxrud, S. Jawahir, J. B. Bender, and S. Sreevatsan. Characterization of methicillin resistant Staphylococcus aureus from bulk tank milk from MN dairy farms. ASM-ESCMID meeting. 2011. K. P. Haran 1, S. M. Godden 1, J. B. Bender 1, S. Sreevatsan. Isolation and Characterization of Methicillin Resistant Staphylococcus aureus from Bulk Tank Milk in Minnesota Dairy Farms. Third International Symposium on Mastitis and Milk Quality. Sept. 22-24, 2011. St. Louis, MO. MISSOURI Ringen DR, Suntrup D, Middleton JR. 2011. Molecular characterization of isolates from pre-partum secretions, teat skin swabs and post-partum milk samples from replacement heifers on a grazing dairy. Proc. 50th Annual Meeting of the Natl. Mast. Counc. Arlington, VA. Jan 23-26, 2010. 127-128. Dank A, Middleton JR. 2011. Efficacy of two teat skin disinfection techniques. Veterinary Research Scholars Symposium. Orlando, Florida. August 2011. TENNESSEE Almeida RA , X Chen, ME Prado, and SP Oliver. 2011. Role of Streptococcus uberis adhesion molecule in adherence to and internalization of Streptococcus uberis into bovine mammary epithelial cells. Third International Symposium on Mastitis and Milk Quality. St. Louis, Missouri USA. Kerro Dego O, RA Almeida, and SP Oliver. 2011. Gene expression of bovine mammary epithelial cells infected with Escherichia coli associated with acute or persistent bovine mastitis. Third International Symposium on Mastitis and Milk Quality. St. Louis, Missouri USA. Kerro Dego Ol, RA Almeida, and SP Oliver. 2011. Identification of virulence-associated genes and their expression patterns in strains of Escherichia coli associated with bovine mastitis. Third International Symposium on Mastitis and Milk Quality. St. Louis, Missouri USA. VIRGINIA Yeiser, E. E., K. E. Leslie and C. S. Petersson-Wolfe. 2011. The effects of flunixin meglumine treatment on activity measures and feed intake during experimentally induced Escherichia coli mastitis. National Mastitis Council Annual Meeting 2011, Arlington, VA. Pg 133-134. Fitzpatrick, C. E., K. E. Leslie, T. F. Duffield, D. F. Kelton, T. J. DeVries and C. S. Petersson-Wolfe. 2011. Objective assessment of pan in dairy cattle with clinical mastitis. National Mastitis Council Annual Meeting 2011, Arlington, VA. Pg 163-164. Neal S.M., W.A. Wark, S.N. Garst, C.S. Petersson-Wolfe, I. Kanevsky-Mullarky. 2011. Deciphering the impact of maternal cells in neonatal health and immune development. Proceedings of the 92nd Annual Conference of Research Workers in Animal Disease.