Brief Summary of Minutes of Annual Meeting

OBJECTIVE 1: Characterize host mechanisms and pathogenic virulence factors associated with mastitis susceptibility and resistance to improve economic outcomes and animal welfare.

Louisiana State University

Member: William E. Owens

Identify and evaluate bacteria causing bovine mastitis in Louisiana. Test bacteria for antimicrobial resistance to mastitis therapeutics.

Ohio State University

Member: Benjamin D. Enger, Kellie M. Enger

An animal trial was completed that investigated how intramammary infections affect mammary gland growth and development in dairy heifers.  First results indicate that IMI reduce the amount of epithelial tissues present in infected mammary glands and increase the abundance of stromal tissues.

University of Minnesota

Members: S. Godden, E. Royster, L. Caixeta, N. Noyes, and B. Crooker

Contributors: Sam Rowe, Pat Gordon, Daryl Nydam

Patel, S. Godden, E. Royster, B. Crooker, T. Johnson, E. Smith, S. Sreevatsan. Prevalence, antibiotic resistance, virulence and genetic diversity of Staphylococcus aureus isolated from bulk tank milk samples of U.S. dairy herds. The objective of this study was to investigate the prevalence, antibiotic resistance gene content, virulence determinants and genomic diversity of S. aureus in bulk tank milk samples (BTM) from U.S. dairy herds. Of 365 pooled BTM samples collected from 189 herds across the United States, S. aureus was cultured from 170 samples, indicating a sample prevalence of 46.6% in the BTM and a herd prevalence of 62.4% (118 out of 189 herds). Among a subset of 138 S. aureus isolates that were stored for further analysis, 124 were genome sequenced after being confirmed as S. aureus using phenotypic tests. Our findings indicated a high prevalence of S. aureus in BTM, with little evidence of antibiotic resistance but often carrying genes for the various enterotoxins. This study identified predominant genetic clones and provides continued surveillance of S. aureus strains found in U.S. dairy herds. Manuscript published in 2021.

Noyes, L.S. Caixeta, B.J. Heins, V.S. Machado, P. Pinedo. Catalyzing an open-community research and education program to leverage the microbiome for the advancement of organic livestock production, using mastitis as a test case. The two objectives of this study are (1) to describe the teat skin microbiome of dairy heifers in certified organic herds in the US and (2) to investigate the association between teat skin microbiome throughout the transition period and the occurrence of intramammary infections. Approximately 500 nulliparous heifers from 5 organic dairy herds in MN, CO, TX, and NM were enrolled in the summer and fall of 2019. Milk culture for the first 5 weeks post-partum for all the cows enrolled in the study was carried out during fall 2019 and spring 2020. Total DNA was extracted and 16S V4 amplicons were sequenced for ~8,000 gauze samples of the external teat apex epithelium and ~2,000 samples of the internal teat canal epithelium. Sequencing of all samples is nearly complete and full data analysis will occur in late 2021 through 2022.

Dean, L.S. Caixeta, N. Noyes, S. Godden, B. Crooker, F. Pena Mosca. Impact of the cow udder microbiome on the biodiversity of milk. Exploratory project to investigate the microbiome in different parts of the mammary gland, and to better understand how microbiome-host interactions influence response to mastitis challenge. Sample collection and sequencing will be completed in early 2022, and data analysis will be completed by end of 2022.

A. Crooker, S. M. Godden, L. Caixeta, A. Seykora, M. Schutz, J. D. Lippolis, J. B. Cole and B. Rosen. Reducing mastitis in the dairy cow by increasing the prevalence of beneficial polymorphisms in genes associated with mastitis resistance. Our premise is that previous selection practices have successfully increased the presence of genetic polymorphisms associated with increased milk yield but have decreased the presences of polymorphisms associated with disease resistance. We are using contemporary Holsteins (CH) and unique unselected Holsteins (UH) that have not been subjected to selection since 1964 to determine differences in immune and inflammatory responses and mammary gene expression. We will link differential phenotypic responses with whole genome sequencing to identify polymorphisms that impact mastitis resistance. These polymorphisms could enhance gene-assisted selection efforts to increase mastitis resistance. Intramammary challenge studies and data analysis are underway. Intramammary challenge studies with Escherichia coli strain P4 and Streptococcus uberis strain 0140J have been conducted and analyses of collected milk, blood and DNA samples are underway. Impact of Holstein genotype on immune response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) was assessed in an ex-vivo whole blood study demonstrated greater cytokine responses in the UH cows.

University of Missouri

Members: John Middleton, Pamela Adkins

Contributors: Samantha Haw

Ongoing collaborations with University of Montreal and University of Calgary to better define the role of non-aureus staphylococci in bovine mastitis. A collaboration with the University of Maryland has been established to evaluate big-data genomics to improve dairy cattle health, including investigating mastitis resistance.

University of Vermont

Members: John Barlow, Feng-Qi Zhao

In collaborations with Drs. Jana Kraft and Thomas McFadden, Zhao lab has investigated that the effects of intramammary challenge of lipopolysaccharide (LPS) on fatty acid profiles in blood and milk. It was shown that LPS challenge significantly changed the fatty acid profiles in triglyceride and phospholipid fractions in blood, but not in milk. (VT MO)

Oregon State University

Member: Massimo Bionaz

Contributors: Serkan Ates, Hunter Ford

Characterize host mechanisms and pathogenic virulence factors associated with mastitis susceptibility and resistance to improve economic outcomes and animal welfare.

Cornell University

Member: Paolo Moroni

Contributors: Gloria Gioia, Anja Sipka, Paul Virkler, Daryl Nydam

We will continue the research program investigating the opportunity to understand the epidemiology of Mycoplasma spp and prevalence in US farms. In collaborations with different group, we investigated genotyping and antimicrobial susceptibility profiling of Streptococcus uberis isolated from a clinical bovine mastitis outbreak in a dairy farm. Molecular characterization contributed to understanding their modes of dissemination, and revealed that Strep. uberis infections could be related to a restricted number of predominant cow-adapted strains.

University of Tennessee

Member: Oudessa Kerro Dego

Contributors: Benti Deresa Gelalcha, Aga Edema Gelgie, Jessica Vidlund, Barbara Gillespie, Ximin Zeng, Jun Lin

Bovine mycoplasmoses, which is mostly caused by Mycoplasma bovis, is a significant problem in the dairy and beef industry. Mycoplasma is an important etiologic agent of bovine mastitis leading to significant economic losses and affecting animal welfare. It is highly contagious and resistant to antibiotics and lacks cell wall rendering certain classes of antibiotics ineffective. Intramammary Mycoplasma bovis infections cause elevation in milk somatic cell count which consequently result in poor milk quality. Current mastitis control measures are not effective against mycoplasma mastitis. In addition, the current lack of information on virulence factors hampered the development of immunological or other alternatives to control M. bovis mastitis. Thus, the lack of preventive measures or treatment options renders infected animal culling as the recommended practice for controlling this disease. The objectives of this study are 1) determine M. bovis IMI status in East TN dairy farms, 2) identify genes responsible for development of M. bovis mastitis using transposon-based mutagenesis and M. bovis transcriptome analysis during early host-M. bovis interactions and 3) Evaluate detailed transcriptomic, histopathological and immunological changes during pathogenesis of experimentally induced M. bovis mastitis.

Summary preliminary results:

• Transposon (Tn4001) mutant library of bovis PG45 were created (Collaborative BARD project with Kimron Veterinary Institute, Israel)
• Mutant clones’ library were characterized for loss of important virulence by in vitro adhesion and invasion assays
• Clones that showed defect to cause in vitro infection of mammary epithelial cells were selected and we will be tested in vivo by experimental challenge infection

University of Idaho

Members: Pedram Rezamand

The Rezamand lab continues to work on identifying relationships between nutrient metabolism during the periparturient period and health issues (mastitis) in Pacific Northwest dairy herds. An objective of the study is to assess the relationship between the milk fatty acid profile of dairy cows with mastitis status of the cows during the periparturient period. 115 healthy and mastitic and lameness cows were randomly selected for milk fatty acid composition (gas chromatography). Health records were categorized based on the occurrence of postpartum diseases. Data were analyzed using linear mixed models in SAS with significance declared at P ≤ 0.05.  results showed that under specific conditions of the experiment, no significant difference in milk fatty acid composition between diseased and healthy cows was observed

OBJECTIVE 2: Assess and apply new technologies that advance mastitis control, milk quality and/or dairy food safety.

University of Minnesota

Members: S. Godden, E. Royster, L. Caixeta, N. Noyes, and B. Crooker

Contributors: Sam Rowe, Pat Gordon, Daryl Nydam

Nusrat Annie Jahan, S. Godden, T. Schoenfus, C. Gebhart, E. Royster, S. Wells, J. Timmerman, and R. Fink. Evaluation of the Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) system in the detection of Mastitis pathogens from Bovine milk samples. In the present study, we prospectively compared MALDI-TOF MS (Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry or MALDI-ToF) to the conventional 16S rDNA sequencing method for the identification of  mastitis isolates (481). Among the 481 milk isolates, 445 (93%) were putatively identified to the genus level by MALDI-TOF MS and 355 (74%) were identified to the species level, but no reliable identification was obtained for 16 (3.3%), and 20 (4.2%) discordant results were identified. In conclusion, our results show that MALDI-TOF MS is a fast and reliable technique which has the potential to replace conventional identification methods for most dairy pathogens, routinely isolated from the milk and dairy products. Manuscript published in 2021.

L.S. Caixeta. Applying precision dairy farming and diagnostic technologies to detect mastitis at the time of and following dry off in dairy cows in a field study. The main objective of this study was evaluate accuracy of precision dairy monitoring and diagnostic technologies (i.e. differential somatic cell counter) in collecting data that effectively identifies appropriate animals for dry cow therapy compared to SCC and bacteriology. In this study, milk samples were collected 1d prior to dry-off and 3 and 5 days after calving. Milk samples were used for the measurement of differential SCC, SCC, and milk culture to identify intramammary infection. Cow enrollment and health and performance was completed between fall 2019 and fall 2020. Data analysis and reported will be completed in 2021.

University of Missouri

Members: John Middleton, Pamela Adkins

Contributors: Samantha Haw

Collaborated with University of Montreal and other NE-1748 stations in the U.S. on work led by Simon Dufour and Daryna Kurban (University of Montreal) to define intramammary infection using MALDI-TOF identification of bacteria isolated from bovine milk samples.  Studied the prevalence of mastitis pathogens in Jersey cattle and their association with subclinical mastitis.

University of Vermont

Members: John Barlow, Feng-Qi Zhao

The Barlow lab collaborated with Dr. De Vliegher and Dr. De Visscher from Ghent University and Dr. Fox from Washington State University in the development of a multilocus sequence strain typing scheme for Staphylococcus chromogenes. (VT, WA, Ghent)

The Barlow lab completed a survey of organic dairy herds in Vermont that identified the frequency of different housing and bedding management systems used for winter housing of lactating cows. Tie-stall barns using wood shavings  bedding was the most common system used on this population of farms.

Utah State University

Members: David Wilson

Contributors: Kerry Rood, Justine Britten         

Screen a large panel of blood and serum clinical pathology tests on dairy calves and develop the most practical reduced panel of tests for prognosis and treatment of sick dairy calves during the first 60 days of life.  This will be compared to a clinical scale of calf disease signs as well as the treatment and disease or health outcomes of the calves.  The goal is to develop an affordable, rapid blood test panel that can be used on calves showing early signs of disease for better prognostic and treatment purposes, especially in large scale calf rearing.

Study different concentrations of a preservative for milk, colostrum and milk replacer fed to calves.  First phase will examine association with bacteria counts over time under the conditions which exist when those products are fed to dairy calves, in comparison with untreated milk.  The goal is to determine a safe but effective concentration of preservative that can be added to colostrum and milk fed to dairy calves.

Cornell University

Member: Paolo Moroni

Contributors: Gloria Gioia, Anja Sipka, Paul Virkler, Daryl Nydam

We will continue to investigate new technology for early identification of Mycoplasma from milk and compare different type of on farm culture. Survey and interviews on the use of antibiotics in New York State farm could be also one of our topics.

University of Tennessee

Member: Oudessa Kerro Dego

Contributors: Benti Deresa Gelalcha, Aga Edema Gelgie, Jessica Vidlund, Barbara Gillespie, Ximin Zeng, Jun Lin

       Prevalence of Antimicrobial Resistant and Extended Spectrum Beta-Lactamase producing Escherichia coli in Dairy Cattle Farms in East Tennessee

Antimicrobials have been widely used in dairy farms to prevent and control dairy cattle diseases since the1960s. This led to the emergence of antimicrobial resistant bacteria (ARB) that, along with their antimicrobial resistance genes (ARGs), can spread from dairy farms to humans. Therefore, regular antimicrobial resistance (AMR) monitoring is important to implement proper mitigation measures. The objective of this study was to determine prevalence of extended-spectrum beta-lactamases producing E. coli (ESBL-E. coli) in in East Tennessee dairy  farms. A total of eighty samples consisting of twenty samples each of bulk tank milk, feces, dairy cattle manure amended soil, and prairie soil adjacent to the farms were collected and cultured for the isolation of E. coli.  Tetracycline (TET^r)-, third-generation cephalosporin (TGC^r)- and nalidixic acid (NAL^r)-resistant E. coli (n=88) were isolated and identified on agar media supplemented with tetracycline, cefotaxime, and nalidixic acid, respectively. TGC^r E. coli were tested for ESBLs and other co-selected ARGs.  

Results: TET^r (74%, n=88) was the most common, followed by TGC^r (20%) and NAL^r (8%). Farms had significant (P<0.001) differences: the highest prevalence of TGC^r (55%) and TET^r (100%) were observed in farm D, while all NAL^r isolates were from farm C. Over 83% of TGC^r isolates (n=18) harbored ESBL gene bla_CTX-M. Majority (78%) of the E. coli isolates were multidrug-resistant (MDR) being positive for beta-lactams (bla_CTX-M), tetracyclines tet(A), tet(B), tet(M)), sulphonamides (sul2), aminoglycosides (strA), and phenicols (floR).

Molecular Epidemiology of Extended-Spectrum β-lactamase Producing E. coli in East Tennessee Dairy Farms

Most dairy farms in the USA- use third-generation cephalosporins (TGC), a medically important class of antibiotics, for the treatment and prevention of infectious diseases of dairy cattle (1). Recent reports are indicating that resistance to a third-generation cephalosporin (TGC) is rising among Enterobacteriaceae such as Escherichia coli (E. coli) in dairy operations and human health settings. We hypothesized that ESBLs-E. coli and -Klebsiella spp.  are widespread in dairy farms and their environments, and they co-carry other resistance genes. The objectives of this study are 1) determine the prevalence of ESBLs-E. coli and -Klebsiella spp.  in dairy cattle and manure, 2) determine the genomic relationship among all ESBLs -E. coli and -Klebsiella spp.

Preliminary results

       A total of 512 rectal fecal samples were collected from 512 cattle from 14 dairy  farms

       A total of 22 bulk tank milk, 25 manure (stored manure, pooled manure from farm floor, lagoon, etc.), 13      

       feed samples, and 17 water samples were collected and screened for ESBL-E. coli and -Klebsiella spp.

       From rectal fecal samples, a total of 278 presumptive ESBLs-E. coli and 208 ESBLs-Klebsiella spp. were   

       isolated and stocked for further confirmation and molecular analysis

       From other farm samples (Bulk Tank Milk, manure, feed and water) 27 presumptive ESBL-E. coli, and 26   

       ESBLs-Klebsiella spp. were isolated and stocked for further confirmation and molecular analysis

       Suspected ESBL-E. coli and -Klebsiella spp. were isolated from all sample types (rectal fecal sample, bulk   

        tank milk, manure, feed, and water).

OBJECTIVE 3: Identify and apply new strategies associated with the control of mastitis that can reduce the use of antibiotics in dairy herds.

Louisiana State University

Member: William E. Owens

Continue evaluation of botanical formulations from plants for antimicrobial activity against mastitis

University of Minnesota

Members: S. Godden, E. Royster, L. Caixeta, N. Noyes, and B. Crooker

Contributors: Sam Rowe, Pat Gordon, Daryl Nydam

S.M. Rowe, S.M. Godden, D.V. Nydam, P.J. Gorden, A. Lago, A.K. Vasquez, E. Royster, J. Timmerman, M.J. Thomas. Randomized controlled non-inferiority trial investigating the effect of two selective dry cow therapy protocols on antibiotic use and udder health: quarter-level outcomes. The objective of this study was to compare culture- and algorithm-guided SDCT programs to blanket dry cow therapy (BDCT) in a multi-site, randomized, natural exposure, non-inferiority trial on quarter-level infection dynamics during the dry period and on cow health and performance during the first 120 DIM. Results showed that the two SDCT approaches (culture- and algorithm-guided) each reduced antibiotic use at dry-off by 55%, without causing any negative impacts on intramammary infection dynamics during the dry period. An economic analysis showed an estimated positive economic return, on average, with either SDCT program. We conclude that SDCT can be used in appropriate U.S. dairy herds to reduce antibiotic use while maintaining udder health.

Godden, S., S. Wells, E. Royster, B. Crooker. Management of Recycled Manure Solids Bedding on Midwest Dairy Farms. Part 1 - Relationship between Processing Method, Udder Health and Production, and Bedding Bacteria Counts. Godden. Our objective was to investigate if recycled manure solids (RMS) processing method was associated with udder health, milk production, and with bedding bacteria counts (BBC) in ready-to-use RMS bedding samples. Twenty-nine premises in MN and WI were enrolled. Processing systems represented included green (GRN; n=7), drum composted (COM; n=4), digested (DIG; n=6) or mechanically dried (DRY; n=12). Farms were visited twice (summer 2019/winter 2020) to collect samples and data. Ready-to-use bedding was cultured to describe BBC. Results showed that herds using DRY or COM RMS processing systems generally had improved udder health and, for DRY, improved milk production, as compared to herds using DIG or GRN solids. Also, BBC were most consistently lowest in ready-to-use DRY or COM samples.  

Godden, S., S. Wells, E. Royster, B. Crooker. Investigation of the Relationship between Method for Processing Recycled Manure Solids and Pathogen Control on Midwest Dairy Farms. Our objectives were to Investigate if method of recycled manure solids (RMS) processing reduces or eliminates viable mastitis pathogens as well as zoonotic pathogens in RMS. In summer of 2021 slurry and pre- and post-processed RMS were sampled from 27 herds in MN and WI.  Laboratory analysis of samples is ongoing.

Godden, S. and E. Royster. Pilot Study: Refining a Culture-Guided Selective Dry Cow Therapy Program (SDCT) to Reduce Antibiotic Use on Dairy Farms. We hypothesize that by identifying and selectively treating only IMI caused by SSLO, we can dramatically reduce antibiotic use at dry-off by as much as 90%, while maintaining udder health, cow health and performance in the subsequent lactation (versus blanket DCT).  In summer, 2020, we conducted a pilot study in 2 commercial herds to evaluate the effect of implementing a SDCT program that identifies and treats only IMI caused by SSLO on measures of quarter health, cow health and performance and antibiotic use (vs. BDCT). Lab culture of samples is ongoing.

Noyes, L. Caixeta, B. Heins, V. Machado, P. Pinedo, A. Formiga, D. Van Nydam, J. Velez, H. Karremann. Catalyzing an open-community research and education program to leverage the microbiome for the advancement of organic livestock production, using mastitis as a test case. The long-term goal of this proposal is to foster application-driven microbiome research activity within the dairy scientific community, thus supporting the discovery and development of novel, evidence-based tools to combat dairy disease and production challenges, with a focus on mastitis. Project objectives described above. 

Rowe, L.S. Caixeta, E. Royster, S. Godden. Clinical trial evaluating the health effects of reducing antibiotic use at dry-off in dairy cows. Randomized clinical trial comparing blanket dry cow therapy and no dry cow therapy in a low input commercial dairy farm. Enrollment of cows started in the summer 2021 and will continue in the spring of 2022. Lactation health and performance for the first 100 DIM will continue until winter 2022. Data analysis and reporting will be completed in 2023.

L.S. Caixeta, S. Godden, W. Knauer, J. Hadrich. G. Cramer. Randomized clinical trial evaluating the effect of administering Bovikalc® dry at dry off on udder health, early lactation health and performance, and economics in dairy cows. Randomized clinical trial investigating the effects of pre dry-off administration of acidogenic bolus on udder health during the dry period and in early lactation. Enrollment of cows was completed in the summer and fall of 2021. Cows will be follow until the spring of 2022. Data analysis and reporting will be completed in 2022.

Oregon State University

Member: Massimo Bionaz

Contributors: Serkan Ates, Hunter Ford

The objective was to assess the potential of using chicory silage in combination with organic selenium in improving the repose to intramammary infection. For the purpose we used 30 Polypay lactating sheep in a 2x2 experimental design where animals were fed for 40 days during pregnancy and 45 days during lactation with isocaloric and isonitrogenous rations containing either chicory silage or grass silage and receiving 0 or 3.6 mg of Se/day as Se-yeast into the concentrate. At 35 days in lactation 2´10⁷ CFU of Strep uberis were infused into both mammary glands. Animals were left untreated for 10 days after which intramammary antibiotics were infused and the sheep dried off. Daily feed and water intake were recorded, milk yield and rectal temperature were measured twice daily, blood was collected prior to and for 10 days after IMI for a complete blood count, leukocyte migration and phagocytosis, and large metabolic, oxidative, liver function, and immune profiling, and milk was collected for milk component analysis, including fatty acid profiling. Preliminary data indicated a minor effect of Se-yeast on the response to mastitis and a more prominent effect of feeding chicory silage.

Ohio State University

Member: Benjamin D. Enger, Kellie M. Enger

An animal trial was completed that investigated the ability of teat sealants to prevent intramammary infections is non lactating heifers. Results indicate that properly administered teat sealants can provide a significant protective effect and are likely to improve first lactation performance given fewer animals begin lactation with infections. Animals with fewer infections are likely to require less antibiotic treatments.

University of Missouri

Members: John Middleton, Pamela Adkins

Contributors: Samantha Haw

Co-authorship on invited reviews led by Herman Barkema and his team at University of Calgary on selective dry cow therapy and selective treatment of clinical mastitis along with authors from other NE-1748 stations in the U.S.A, Canada, and Europe.

Cornell University

Member: Paolo Moroni

Contributors: Gloria Gioia, Anja Sipka, Paul Virkler, Daryl Nydam

Co-authorship on invited reviews led by Sandra Godden  and his team at University of Minnesota on selective dry cow therapy and selective treatment of clinical mastitis. Validation of commercial PCR for identification of Mycoplasma spp and Staphylococcus aureus

University of Tennessee

Member: Oudessa Kerro Dego

Contributors: Benti Deresa Gelalcha, Aga Edema Gelgie, Jessica Vidlund, Barbara Gillespie, Ximin Zeng, Jun Lin

University of Tennessee

Member: Oudessa Kerro Dego

Contributors: Benti Deresa Gelalcha, Aga Edema Gelgie, Jessica Vidlund, Barbara Gillespie, Ximin Zeng, Jun Lin

      Evaluation of Efficacy of Staphylococcal Surface Proteins Vaccine to Control Mastitis in Dairy Cows

Bovine staphylococcal mastitis is a major cause of economic losses in the dairy industry throughout the world. Coagulase-positive Staphylococcus aureus is the most common pathogen that causes mastitis in dairy cows. Staphylococcus chromogenes is one of coagulase-negative staphylococci frequently isolated from intramammary infection (IMI) characterized by high somatic cell count (SCC). Sustainable alternatives to antimicrobials such as improved effective vaccines are critically required to control mastitis.

We developed Staphylococcus aureus surface proteins (SASP) and Staphylococcus chromogenes surface proteins (SCSP) based vaccines. Previously, we completed two evaluations of the protective efficacy of these vaccines using controlled experimental vaccination and challenge studies. Our results showed that both SASP and SCSP vaccines are immunogenic and the induced immunity partially protected vaccinated cows from S. aureus mastitis.  In the current study, we evaluated the immunological responses and protection from mastitis in dairy cows vaccinated with  SCSP and  SASP vaccines over one full lactation of 300 days. A total of 45 dairy cows in their 1^st or 2^nd lactation from the East Tennessee Research and Education Center dairy herd were randomly enrolled in vaccination with SCSP (n=16) or SASP (n=15) with Emulsigin-D® adjuvant (Phibro Animal Health)  at the drying off (D0), 21, and 42 days after drying off (D21 and D42). Control cows (n=14) were injected with PBS (pH 7.4) mixed with Emulsigin-D® at similar time points. Cows in the SCSP and SASP groups were vaccinated with 1.2 mg of SCSP and SASP vaccine, respectively. All cows were vaccinated subcutaneously (SC) in the neck area. The 1st dose was administered on the left side of the neck, the 2nd dose on the right side of the neck, and the 3rd dose on the lower left side of the neck.Serum and milk antibody titers (IgG, IgG1, IgG2, and IgA) induced by SCSP or SASP were measured by ELISA. We monitored them for antibody titers in milk and serum, mastitis status (clinical, subclinical, no mastitis), somatic cell count (SCC), milk yield, bacterial count in CFU/mL of milk over one full lactation cycle.

Results: Of the total 45 cows, 9 were removed from the study due to non-vaccine-related complications. All vaccinated cows had no allergic reactions or vaccine-related complications, so these vaccines are safe. Similar to the previous two studies, SASP and SCSP vaccines induced significantly increased antibody titers. The subsequent experimental challenge results showed that both vaccines confer partial protection, but SCSP conferred better protection than SASP. The duration of induced immunity was about 4 months. A total of 4  vaccine injections are required throughout lactation periods at drying off (D0), at days 21 and 42 after drying off, and at 120 days in milk or 120 days after calving.

Evaluation of Immunogenicity of Enterobactin Conjugate Vaccine for the Control of E. coli Mastitis in Dairy Cows

Coliform bacteria, such as Escherichia coli, Klebsiella, and Enterobacter, widely exist in the environment and are common causes of clinical mastitis in dairy cows. Escherichia coli is the most common species isolated in more than 80% of cases of coliform mastitis. Despite decades of vaccine trials to develop an effective vaccine against E. coli mastitis in dairy cows, the efficacy of the current J5 based vaccine is limited only to reducing clinical disease and production losses. The limited effectiveness is probably due to antibody inaccessibility and antigenic diversity/variation issues. Therefore, a novel vaccine, with a new target of less antigenic changes, is highly warranted. Recently, a novel essential nutrient deprivation-based immunization approach, which targets Enterobactin (Ent), has been under investigation.

The objective of this study was to determine Ent-specific immune response to Ent-KLH vaccine in dairy cows during early lactation. A total of 12 healthy Holstein dairy cows from East Tennessee Research and Education Center Little River Animal and Environmental Unit Herd (ETREC-LAEU) were divided into two groups of 6 cows each. Cows in group 1 were vaccinated subcutaneously with 200 μg Ent-KLH vaccine with Emulsigen-D (first vaccination) and Freund’s incomplete adjuvant (2^nd and 3^rd vaccination) in the total volume of 5 mL. Cows in group 2 were injected with PBS (pH7.4) with Emulsigen-D (first injection) and Freund’s incomplete adjuvant (2^nd and 3^rd injections). The serum and milk samples were analyzed for the Ent-specific IgG1, IgG2, and IgA immune responses by indirect ELISA.

Results: KLH-Ent vaccine induced significantly higher antibody in serum and milk of vaccinates compared with control cows. In particular, in vaccinated cows, the IgG and IgG2 response against KLH-Ent conjugate and Ent reached the peak at calving (C), and continued increasing until 30 days after calving (C30). Ent specific IgG1 and IgA peaked at D42 and remained high at C and C14, and then quickly declined. The KLH-Ent successfully triggered strong Ent-specific immune response in dairy cows without significantly affecting the microbiota diversity and gut health. Thus, Ent conjugate vaccine may serve as effective vaccine against E. coli mastitis in dairy cows.

Developing a universal nanoparticle-based vaccine delivery system against infectious diseases (collaborative project between Dr. Paul Dalhaimer from Department of Chemical and Biomolecular Engineering, Tickle College of Engineering, UTK, Knoxville and Dr. Oudessa Kerro Dego, Department of Animal Science, UTIA, Knoxville). The objective of this study was to develop nanoparticles that have antigens of interest on the same nanoparticle. we propose to use the spherical nanoparticles (SNPs), diblock copolymers: poly(ethylene-oxide)-b-poly(butadiene) (PEO-b-PBD). PEO is FDA-approved for use in the body. Current nanoparticles tend to have just one functionality group – usually the antigen. Our nanoparticles have many attributes that make them ideal for use as vaccines. First, they are made from benign polymers that have been extensively administered to rodents and shown low toxicity. Second, they are stable even at temperatures ~80ºC. This is a very important requirement for vaccines to be used in refrigeration-poor countries. Third, they have ideal physical dimensions. They are ~50 nm in diameter, which is ideal for reaching the lymph system from site of administration.  Our nanoparticle allows for the attachment of multiple combinations of antigens and targeting peptides which makes it highly advantageous because you can attach multiple antigens and targeting peptides on one nanoparticle.

Summary of preliminary results:

We constructed nanoparticle-Staphylococcus aureus surface protein peptide (SASP-peptide) Vaccinated two groups of mice with Nanopartilce alone (Group 1) and nanoparticle-attached to SASP- peptide at days 0, 14 and 28. Blood samples were collected at days 0, 14, 28 before each vaccination and at day 42  two weeks after last vaccination.We measured induced immune responses (IgG, IgM and IgA) against nanoparticle and nanoparticle-SASP-peptide conjugate by ELISA. Serum  IgG antibody specific to  nanoparticle-SASP peptide increased  significantly from base line at D28 and D42. Both Nanoparticle alone and Nanoparticle-SASP peptide induced increased IgM antibody in serum at D28. We are currently running second vaccination and immune response evaluation in mice.

WORK PLANNED FOR THE COMING YEAR, LISTED BY OBJECTIVE:

OBJECTIVE 1

University of Missouri

Members: John Middleton, Pamela Adkins

Contributors: Samantha Haw

We will be investigating the antibacterial activity of Staphylococcus chromogenes isolates originating from dairy cattle, specifically looking at their in vitro ability to inhibit growth of Gram-positive mastitis pathogens. We will also begin a collaborative project with the University of Vermont, working to do whole genome sequencing and strain typing of S. chromogenes isolates, looking to determine genetic determinants of phenotypic behavior. 

Louisiana State University

Member: William E. Owens

Identify and evaluate bacteria causing bovine mastitis in Louisiana. Test bacteria for antimicrobial resistance to mastitis therapeutics.

University of Minnesota

Members: S. Godden, E. Royster, L. Caixeta, N. Noyes, and B. Crooker

Contributors: Sam Rowe, Pat Gordon, Daryl NydamA. Crooker, S. M. Godden, L. Caixeta, A. Seykora, M. Schutz, J. D. Lippolis and B. Rosen. Reducing mastitis in the dairy cow by increasing the prevalence of beneficial polymorphisms in genes associated with mastitis resistance. Analysis of collected milk, blood and DNA samples from unselected and contemporary Holsteins subjected to intramammary challenges of Escherichia coli strain P4 or Streptococcus uberis strain 0140J will continue. Impact of Holstein genotype on immune response to gram negative and gram positive bacteria that commonly cause mastitis will be assessed using ex-vivo incubation models.

Caixeta, S. Dow, N. Noyes, B. Crooker, S. Godden, D. Nydam, B. Walcheck. Effect of mucosal immune stimulation on mammary gland immune responses during the dry period and early lactation period in dairy cows. The two objectives of this project are to determine the effect of MucosImmune on (1) the innate immune response in the mammary gland during the period immediately before and after parturition, and (2) the resistance to bacterial infection of the mammary gland. Initial dose titration experiment will be carried out in the spring of 2022. The three main experiments proposed in this project will be completed between the summer of 2022 and winter 2023.

University of Vermont

Members: John Barlow, Feng-Qi Zhao

Zhao lab will study the effects of LPS and different cytokines on milk synthesis in mammary epithelia cells.

Ohio State University

Member: Benjamin D. Enger, Kellie M. Enger

Continue to investigate how mammary growth is altered by mastitis by assessing proliferation and apoptosis of epithelial cells in heifer mammary gland tissues and also assess how myoepithelial cells and their migration into the fat pad are affected by inflammatory processes.

Cornell University

Member: Paolo Moroni

Contributors: Gloria Gioia, Anja Sipka, Paul Virkler, Daryl Nydam

We will be investigating the antibacterial activity of Lactococcus garvieae  isolates originating from milk bedding and water.

University of Idaho

Members: Pedram Rezamand

We will continue to examine the relationship between nutrient metabolism and periparturient diseases (i.e., mastitis). 

OBJECTIVE 2

University of Minnesota

Members: S. Godden, E. Royster, L. Caixeta, N. Noyes, and B. Crooker

Contributors: Sam Rowe, Pat Gordon, Daryl Nydam

Godden, S., S. Wells, E. Royster, B. Crooker. Management of Recycled Manure Solids Bedding on Midwest Dairy Farms. Part 1 - Relationship between Processing Method, Udder Health and Production, and Bedding Bacteria Counts.  Final results will be analyzed and manuscripts submitted in 2022.  

Godden, S., S. Wells, E. Royster, B. Crooker. Investigation of the Relationship between Method for Processing Recycled Manure Solids and Pathogen Control on Midwest Dairy Farms. Our objectives were to Investigate if method of recycled manure solids (RMS) processing reduces or eliminates viable mastitis pathogens as well as zoonotic pathogens in RMS. In summer of 2021 slurry and pre- and post-processed RMS were sampled from 27 herds in MN and WI.  Laboratory analysis of samples is ongoing. Data analysis and reporting will occur in 2022.

Oregon State University

Member: Massimo Bionaz

Contributors: Serkan Ates, Hunter Ford

Complete the analyses including blood profiling and milk fatty acid profiling and submit two manuscripts to peer-reviewed journals

University of Tennessee

Member: Oudessa Kerro Dego

Contributors: Benti Deresa Gelalcha, Aga Edema Gelgie, Jessica Vidlund, Barbara Gillespie, Ximin Zeng, Jun Lin

Molecular Epidemiology of Extended-Spectrum β-lactamase Producing E. coli in East Tennessee Dairy Farms

Identification of Virulence Factors & Pathogenesis of Mycoplasma bovis Mastitis (MBM) in Dairy

University of Vermont

Members: John Barlow, Feng-Qi Zhao

The Barlow lab will collaborate with Dr. Pamela Adkins at the University of Missouri to explore the epidemiology and pathogenesis of Staphylococcus chromogenes intramammary infections. The Barlow lab will complete whole genome sequencing of Staphylococcus species isolates to identify potential virulence factors and antimicrobial resistance genes, and explore the phylogeny and ecology of Staphylococcus species and mobile genetic elements among these species. The Barlow lab will continue to explore the epidemiology of Staphylococcus aureus and non-aureus staphylococci in small to medium sized dairy farms.

Cornell University

Member: Paolo Moroni

Contributors: Gloria Gioia, Anja Sipka, Paul Virkler, Daryl Nydam

Investigation of the Relationship between different type of bedding and control of mastitis.

OBJECTIVE 3

University of Vermont

Members: John Barlow, Feng-Qi Zhao

The Barlow lab will continue to explore the relationship between housing and bedding management practices and mastitis risk using culture-based and culture independent (metagenomic) methods. Barlow lab will continue to explore the potential role of endogenous inhibitor bacteria and bacteriocins in the epidemiology of Staphylococcus mastitis. We will use in vitro co-culture systems and metagenomic methods to describe bacterial factors influencing colonization and infection of mammary glands.

University of Tennessee

Member: Oudessa Kerro Dego

Contributors: Benti Deresa Gelalcha, Aga Edema Gelgie, Jessica Vidlund, Barbara Gillespie, Ximin Zeng, Jun Lin

Optimization of vaccination regimen for staphylocococcal, streptococcal and E. coli vaccines:

Evaluation of Efficacy of Staphylococcal and Streptococcal Surface Proteins Vaccine to Control Mastitis in Dairy Cows

Evaluation of Immunogenicity of Enterobactin Conjugate Vaccine for the Control of E. coli Mastitis in Dairy Cows

University of Minnesota

Members: S. Godden, E. Royster, L. Caixeta, N. Noyes, and B. Crooker

Contributors: Sam Rowe, Pat Gordon, Daryl Nydam

Godden, S. and E. Royster. Pilot Study: Refining a Culture-Guided Selective Dry Cow Therapy Program (SDCT) to Reduce Antibiotic Use on Dairy Farms. Data analysis and reporting will occur in 2022.

Louisiana State University

Member: William E. Owens

Continue evaluation of botanical formulations from plants for antimicrobial activity against mastitis pathogens.

PUBLICATION LIST: 

Peer-Reviewed Literature

Ohio State University

L.R. Larsen, P.H. Baker, K.M. Enger, L.E. Moraes, P.R.F. Adkins, J.A. Pempek, C.A. Zimmerly, S.M. Gauta, R.L. Bond and B.D. Enger. Administration of internal teat sealant in primigravid dairy heifers at different times of gestation to prevent intramammary infections at calving. J. Dairy Sci. 104: 12773-12784.

Enger, B.D. and J. R. Middleton. 2021. Letter to the Editor: Comments on “Mammary microbial dysbiosis leads to the zoonosis of bovine mastitis: a One-Health perspective” by Maity and Ambatipudi. FEMS Microbiol. Ecol. 97:fiab076.

Enger, K.M., N.R. Hardy, E.M. Hist, and B.D. Enger. 2021. Relationship between intramammary infection and antibody concentrations in Jersey and Holstein colostrum. J. Dairy Sci. 104:6124-6133.

Hardy, N.R., K.M. Enger, J. Hanson, M.L. Eastridge, L.E. Moraes, and B.D. Enger. 2021. Organization of mammary blood vessels as affected by mammary parenchymal region and estradiol administration in Holstein heifer calves. J. Dairy Sci. 104:6200-6211.

University of Tennessee

Kerro Dego O, Almeida R, Ivey S, Agga GE. 2021. Evaluation of Streptococcus uberis Surface Proteins as Vaccine Antigens to Control S. uberis Mastitis in Dairy Cows. MDPI Vaccines 9:868.  https://doi.org/10.3390/vaccines9080868

Abdi RD, Gillespie BE, Ivey S, Pighetti GM, Almeida RA, Kerro Dego O. 2021. Antimicrobial Resistance of Major Bacterial Pathogens from Dairy Cows with High Somatic Cell Count and Clinical Mastitis. Animals (Basel) 11. DOI: 10.3390/ani11010131

Balemi A, Gumi B, Amenu K, Girma S, Gebru M, Tekle M, Rius AA, D'Souza DH, Agga GE, Kerro Dego O. 2021. Prevalence of Mastitis and Antibiotic Resistance of Bacterial Isolates from CMT Positive Milk Samples Obtained from Dairy Cows, Camels, and Goats in Two Pastoral Districts in Southern Ethiopia. Animals (Basel) 11.DOI: 10.3390/ani11061530

Gelalcha, B. D., D. B. Ensermu, G. E. Agga, M. Vancuren, B. E. Gillespie, D. H. D’Souza, C. Okafor, and O. Kerro Dego. 2021. Prevalence of Antimicrobial Resistant and Extended Spectrum Beta-Lactamase producing Escherichia coli in Dairy Cattle Farms in East Tennessee, Foodborne pathogens and disease, In press.

Gelalcha, B. D., S. Brown, H. Crocker, G. E. Agga, and O. Kerro Dego. 2021. Regulation Mechanisms of Virulence Genes in Enterohemorrhagic Escherichia coli (EHEC): A review, Foodborne pathogens and disease, In press.

University of Minnesota

Fernandes LM, Guimaraes I, Noyes NR, Caixeta L, Machado, V. Impact of subclinical mastitis detected in the first month of lactation on somatic cell count linear scores, milk yield, fertility, and culling of dairy cows in certified organic herds. Journal of Dairy Science. February 2021. https://doi.org/10.3168/jds.2020-19153

Rowe, S. , S. Godden, E. Royster, J. Timmerman, M. Boyle. 2021. Post-calving udder health and productivity in cows approaching dry-off with intramammary infections caused by non-aureus Staphylococcus, Aerococcus, Enterococcus, Lactococcus and Streptococcus species. J. Dairy Sci. 104(5) 6061-6079

Rowe, S. , A. Vasquez, S. Godden, D. Nydam, E. Royster, J. Timmerman, M. Boyle. 2021. Evaluation of four predictive algorithms for intramammary infection status in late lactation cows. J. Dairy Sci. 104(10): 11035-11046.

Rowe, S. , D. Nydam, S. Godden, P. Gorden, A. Lago, A. Vasquez, E. Royster, J. Timmerman, M. Thomas, R. Lynch. 2021. Partial budget analysis of culture and algorithm guided selective dry cow therapy. J. Dairy Sci. 104:5652–5664  https://doi.org/10.3168/jds.2020-19366

Patel, K., S. Godden, E. Royster, B. Crooker, T. Johnson, E. Smith, S. Sreevatsan. 2021. Prevalence, antibiotic resistance, virulence and genetic diversity of Staphylococcus aureus isolated from bulk tank milk samples of U.S. dairy herds. BMC Genomics 22:367https://doi.org/10.1186/s12864-021-07603-4

Jahan, N. , S. Godden, E. Royster, T. Schoenfuss, C. Gebhart, J. Timmerman and R. Fink. 2021. Evaluation of the Matrix-Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) System in the Detection of Mastitis Pathogens from Bovine Milk Samples. J. Microbiol Methods. 182: 106168. https://doi.org/10.1016/j.mimet.2021.106168

Lippolis, J. D., E. J. Putz, T. A. Reinhardt, E. Casas, W. J. Weber and B. A. Crooker. Submitted. Effect of Holstein genotype on immune response to an intramammary Escherichia. coli challenge. Submitted. J. Dairy Sci. JDS.2021-21166. Under revision.

University of Missouri

DeBuck J, Ha V, Naushad S, Nobrega DB, Luby C, Middleton JR, De Vliegher S, Barkema HW.  2021.  Non-aureus staphylococci and udder health.  Front Vet Sci.  April 15;8:658031. Invited Review.  https://doi.org/10.3389/fvets.2021.658031  

Enger BD, Middleton JR.  2021.  Letter to the Editor – Reply to the article: “Mammary microbial dysbiosis leads to the zoonosis of bovine mastitis: a One-Health perspective” FEMS Microbiology Ecology.  2021 Jun 9: https://doi.org/10.1093/femsec/fiab076

University of Vermont

Miyuki Takashima, Christian Lalonde, Laura Olszanski, and Feng-Qi Zhao. (2021). Localized and systemic inflammation mediators in a murine acute mastitis model. Journal of Inflammation Research 14:4053–4067. doi: 10.2147/JIR.S313799

Andrews T, Jeffrey Cc, Gilker R, Neher D, Barlow JWa. 2021. Survey design and implementation quantifies winter housing and bedding types used on Vermont organic dairy farms. J. Dairy Science Apr 14;S0022-0302(21)00517-8 DOI: 10.3168/jds.2020-19832

Huebner R, Mugabi Rc, Hetesy Gd, Fox L, De Vliegher S, De Visscher A, Barlow JWa, Sensabaugh G. 2021. Characterization of genetic diversity and population structure within Staphylococcus chromogenes by multilocus sequence typing. PLoS One. Mar 15; 6(3): e0243688. https://www.biorxiv.org/content/10.1101/2020.11.30.403683v1 DOI: 10.1371/journal.pone.0243688

Cornell University

Hoekstra J, Zomer AL, Rutten VPMG, Benedictus L, Stegeman A, Spaninks MP, Bennedsgaard TW, Biggs A, De Vliegher S, Mateo DH, Huber-Schlenstedt R, Katholm J, Kovács P, Krömker V, Lequeux G, Moroni P, Pinho L, Smulski S, Supré K, Swinkels JM, Holmes MA, Lam TJGM, Koop G. Genomic analysis of European bovine Staphylococcus aureus from clinical versus subclinical mastitis. Sci Rep. 2020 Oct 23;10(1):18172. doi: 10.1038/s41598-020-75179-2. PMID: 33097797; PMCID: PMC7584570.

Kumar R, Register K, Christopher-Hennings J, Moroni P, Gioia G, Garcia- Fernandez N, Nelson J, Jelinski MD, Lysnyansky I, Bayles D, Alt D, Scaria J. Population Genomic Analysis of Mycoplasma bovis Elucidates Geographical Variations and Genes associated with Host-Types. Microorganisms. 2020 Oct 10;8(10):1561. doi: 10.3390/microorganisms8101561. PMID: 33050495; PMCID: PMC7650767.

Gioia G, Addis MF, Santisteban C, Gross B, Nydam DV, Sipka AS, Virkler PD, Watters RD, Wieland M, Zurakowski MJ, Moroni P. Mycoplasma species isolated from bovine milk collected from US dairy herds between 2016 and 2019. J Dairy Sci. 2021 Apr;104(4):4813-4821. doi: 10.3168/jds.2020-19171. Epub 2021 Feb 19. PMID: 33612245.

Wieland M, Geary CM, Gioia G, Case KL, Moroni P, Sipka A. Vacuum Dynamics as an Alternative Method for Detection of Bimodal Milk Ejection in Dairy Cows. Animals (Basel). 2021 Jun 23;11(7):1860. doi: 10.3390/ani11071860. PMID: 34201426; PMCID: PMC8300128.

Gioia G, Addis MF, Goodman LB, Mitchell PK, Thompson B, Goodrich E, Moroni P. Draft Genome Sequence of Acholeplasma laidlawii Isolated from the Conjunctiva of a Heifer with Infectious Bovine Keratoconjunctivitis. Microbiol Resour Announc. 2021 Jan 28;10(4):e01345-20. doi: 10.1128/MRA.01345-20. PMID: 33509992; PMCID: PMC7844077.

Bulut E, Stout A, Wemette M, Llanos-Soto S, Schell RC, Greiner Safi A, Shapiro MA, Moroni P, Ivanek R. How does public perception of antibiotic use on dairy farms contribute to self-reported purchasing of organic? J Food Sci. 2021 May;86(5):2045-2060. doi: 10.1111/1750-3841.15720. Epub 2021 May 6. PMID: 33955540; PMCID: PMC8251749.

Monistero V, Barberio A, Cremonesi P, Castiglioni B, Morandi S, Lassen DCK, Astrup LB, Locatelli C, Piccinini R, Addis MF, Bronzo V, Moroni P. Genotyping and Antimicrobial Susceptibility Profiling of <i>Streptococcus uberis</i> Isolated from a Clinical Bovine Mastitis Outbreak in a Dairy Farm. Antibiotics (Basel). 2021 May 28;10(6):644. doi: 10.3390/antibiotics10060644. PMID: 34071296; PMCID: PMC8229259.

Mann S, Curone G, Chandler TL, Moroni P, Cha J, Bhawal R, Zhang S. Heat treatment of bovine colostrum: I. Effects on bacterial and somatic cell counts, immunoglobulin, insulin, and IGF-I concentrations, as well as the colostrum proteome. J Dairy Sci. 2020 Oct;103(10):9368-9383. doi: 10.3168/jds.2020-18618. Epub 2020 Aug 20. PMID: 32828510.

Llanos-Soto SG, Vezeau N, Wemette M, Bulut E, Greiner Safi A, Moroni P, Shapiro MA, Ivanek R. Survey of perceptions and attitudes of an international group of veterinarians regarding antibiotic use and resistance on dairy cattle farms. Prev Vet Med. 2021 Mar;188:105253. doi: 10.1016/j.prevetmed.2020.105253. Epub 2021 Jan 15. PMID: 33524793.

Penati M, Sala G, Biscarini F, Boccardo A, Bronzo V, Castiglioni B, Cremonesi P, Moroni P, Pravettoni D, Addis MF. Feeding Pre-weaned Calves With Waste Milk Containing Antibiotic Residues Is Related to a Higher Incidence of Diarrhea and Alterations in the Fecal Microbiota. Front Vet Sci. 2021 Jul 8;8:650150. doi: 10.3389/fvets.2021.650150. PMID: 34307516; PMCID: PMC8298036.

Capoferri R, Cremonesi P, Castiglioni B, Pisoni G, Roccabianca P, Riva F, Filipe J, Del Corvo M, Stella A, Williams JL, Rupp R, Moroni P. Comparison of the response of mammary gland tissue from two divergent lines of goat with high and low milk somatic cell scores to an experimental Staphylococcus aureus

infection. Vet Immunol Immunopathol. 2021 Apr;234:110208. doi: 10.1016/j.vetimm.2021.110208. Epub 2021 Feb 19. PMID: 33640660.

Utah State University

Britten JE, Rood KA, Wilson DJ: Comparison of bovine mammary involution and intramammary infections following intramammary treatment with casein hydrolysate and other conventional treatments at dry-off. Animals 11:8:1-12, 2021

Abstracts

Oregon State University

Hunter Ford, Massimo Bionaz, Serkan Ates, Joe Klopfenstein, Jorge Vanegas, Gracia Puerto Hernandez, Callan Stowell, Paige Tafoya, Michele McCann, Mustafa Bozkus, 64 Feeding Chicory-plantain Silage and Se-yeast on Lactating Ewe Subjected to Intramammary Infection: Effect on Performance and Milk Quality, Journal of Animal Science, Volume 99, Issue Supplement_3, November 2021, Pages 34–35, https://doi.org/10.1093/jas/skab235.060

Hunter Ford, Massimo Bionaz, Serkan Ates, Joe Klopfenstein, Jorge Vanegas, Sebastiano Busato, Daniella Hasan, Shelby Park, Carolyn Pearce, Tyler Quinones, Kate Ness, Mariangel Aleman Torres, 62 Feeding Chicory-plantain Silage and Se-yeast to Lactating Ewes Subjected to Intramammary Infection: Effect on the Immune System, Journal of Animal Science, Volume 99, Issue Supplement_3, November 2021, Pages 32–33, https://doi.org/10.1093/jas/skab235.056

Ohio State University

Enger, B.D., P.H. Baker, M.A. McGuire, R.M. Akers, S.K. Jacobi, and K.M. Enger. 2021. Effect of intramammary infections on heifer mammary gland growth and development. Conference of Research Workers in Animal Disease Proceedings:244

Larsen, L.R., P.H. Baker, K.M. Enger, L.E. Moraes, and B.D. Enger. 2021. Intramammary infection prevalence and secretion characteristics of primigravid dairy heifers. J. Dairy Sci. 104 (Suppl. 1):256.

Larsen, L.R., P.H. Baker, K.M. Enger, L.E. Moraes, and B.D. Enger. 2021. Earlier administration of an internal teat sealant in primigravid dairy heifers to prevent intramammary infections at calving. J. Dairy Sci. 104 (Suppl. 1):15.

Rodriguez, M., W.P. Weiss, B.D. Enger, K. Lee, and C. Lee. 2021. Effects of different levels of vitamin A supply on production and blood parameters of transition cows. J. Dairy Sci. 104 (Suppl. 1):310.

Larsen, L.R., P.H. Baker, K.M. Enger, L.E. Moraes, and B.D. Enger. 2021. Earlier administration of internal teat sealant in dairy heifers to prevent intramammary infections at calving and lessen potential impacts on performance. Proc. 2021 Tri-State Nutrition Conference:169.

University of Minnesota

Rowe, S.M., M. Dziuba, B. Boyum, S. Godden, E. Royster, L. Caixeta. 2021. Negatively Controlled Trial Investigating the Effect of Dry Cow Therapy on Clinical Mastitis and Culling. ADSA /Annu Mtg. (Virtual) July 11-14, 2021.

Rowe, S.M., S.M. Godden, D.V. Nydam, A. Vasquez. 2021. Selective dry-cow therapy protocols to enhance udder health from dry-off to calving. ADSA Annu Mtg. July 11-14, 2021.

Brink, A.A., W.J. Weber, J.D. Lippolis, J.B. Cole, S.M. Godden, P.A. Crooker. Effect of Holstein milk yield genotype on ex-vivo innate immune response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) during the periparturient period. ADSA Annu Mtg (virtual). July 11-14, 2021.

Rowe, S.M., A. Vasquez, S.M. Godden, D.V. Nydam, E. Royster, J. Timmerman, M. Boyle. Evaluation of four predictivealgorithms for intramammmary infection status in late lactation cows. Proc. 60^th Annu. Conf National Mastitis Council. Jan. 25-28, 2021 (virtual). 

Rowe, S.M., D.V. Nydam, S.M. Godden, P. Gorden, A. Lago., E. Royster, A. Vasquez, M. Ghomas, R. Lynch. Partial budget analysis of selective dry cow strategies. Proc. 60^th Annu. Conf National Mastitis Council. Jan. 25-28, 2021 (virtual).

Vasquez, D.V. Nydam, S.M. Rowe, S.M. Godden, P. Gorden, A. Lago, E. Royster, M. Thomas, R. Lynch. Impact of dry cow antimicrobial use on the milk microbiome: Caracterization of dry-off and fresh-cow samples from 6 U.S. dairy farms. Proc. 60^th Annu. Conf National Mastitis Council. Jan. 25-28, 2021 (virtual).

Godden, S., F. Peña Mosca, E. Royster, B. Crooker, P. Raynor, and K. Janni. 2021. Management of Recycled Manure Solids Bedding on Midwest Dairy Farms. Part 1 - Relationship between Processing Method, Udder Health and Production, and Bedding Bacteria Counts. Proc. 60^th Annu. Conf National Mastitis Council. Jan. 25-28, 2021 (virtual).

Peña Mosca, F., S. Godden, E. Royster, B. Crooker, P. Raynor, and K. Janni. 2021. Management of Recycled Manure Solids Bedding on Midwest Dairy Farms. Part 2 - Relationship between Processing Method, Bedding Characteristics and Bedding Bacterial Count. Proc 60^th Annu. Conf National Mastitis Council. Jan. 25-28, 2021 (virtual).

Brink, A. A., W. J. Weber, J. D. Lippolis, J. B. Cole, S. M. Godden and B. A. Crooker. 2021. Effect of Holstein milk yield genotype on ex-vivo innate immune response to lipopolysaccharide (LPS) and lipoteichoic acid (LTA) during the periparturient period. ADSA Annual Meeting. J. Dairy Sci. July 11-14. (virtual).

University of Missouri

Kurban D, Roy JP, Kabera F, Frechette A, Um MM, Albaaj A, Rowe S, Godden S, Adkins PRF, Middleton JR, Gauthier ML, Keefe G, DeVries TJ, Kelton DF, Moroni P, Veiga dos Santos M, Barkema HW, Dufour S.  2020.  Diagnosing intramammary infection: a scoping review and meta-analysis on frequency and udder health relevance of microorganism species retrieved in bovine milk samples.  Journée de recherché, Faculty of Veterinary Medicine, University of Montreal, Quebec, Canada.  15-18 March 2021.

Resendiz-Biermaier K5, Ringen DR, Middleton JR.  2021.  Epidemiology of staphylococcal mastitis in primiparous heifers on a pasture-based dairy.  Proceedings of the 43rd Annual CVM Research Day. May 7, 2021.  Abstract #48.Adkins PRF. Heat stress and the microbiome of dairy cows. American College of Veterinary Internal Medicine Annual Forum, June, 2021

University of Tennessee

Gelalcha, B. D., B. Gillespie and O. Kerro Dego. Extended-Spectrum Beta-Lactamases Producing E. coli in Dairy Cattle Farms, MRW 2021  annual meeting Nov. 3 – 4, Virtual.

Gelgie, A. E., R. Almeida and O. Kerro Dego. 2021. Identification of Virulence Factors and Pathogenesis of Mycoplasma bovis Mastitis in Dairy Cattle, A Proposal, MRW 2021  Annual meeting Nov. 3 – 4, Virtual.

Vidlund, J. B. Gillespie and o. Kerro Dego. 2021. Efficacy of Staphylococcal surface associated protein Vaccines against Mastitis. MRW 2021  Annual meeting Nov. 3 – 4, Virtual.

Kerro Dego, O., X. Zeng, J. Vidlund, B. Gillespie, L. Cao, H. Wang and J. Lin. 2021. Evaluation of Immunogenicity of Enterobactin Conjugate Vaccine for Controlling E. coli Mastitis in Dairy Cows, MRW 2021  Annual meeting Nov. 3 – 4, Virtual.

Cornell University

Addis F., Pisanu S., Penati M., Monistero V., Gazzola A., Castiglioni B., Cremonesi P., Moroni P., Pagnozzi D., and R. Piccinini. Comparative Secretome Analysis of Staphylococcus aureus Strains Belonging to Sequence Types with Different Within-herd Mastitis Prevalence Proceeding of the 60^nd National Mastitis Council Annual Meeting pp.90-91. January 26-28, 2021 Virtual.

Monistero V., Cremonesi P., Morandi S., Barberio A., Castiglioni B., Locatelli C., Piccinini R., and P. Moroni.  Molecular Characterization of Streptococcus uberis Strains Isolated from a Clinical Bovine Mastitis Outbreak in an Italian Dairy Farm. Proceeding of the 60^nd National Mastitis Council Annual Meeting pp.122-123. January 26-28, 2021 Virtual.

Penati M., Sala G., Biscarini F., Boccardo A., Bronzo V., Castiglioni B., Cremonesi P., Pravettoni D., Locatelli C., Moroni P., and M. F. Addis. Waste Milk Containing Antimicrobials Affects Gut Health and Microbiota Diversity in Calves. Proceeding of the 60^nd National Mastitis Council Annual Meeting pp.124-125. January 26-28, 2021 Virtual.

University of Idaho

Tsai C. Y., H. H. Hung, T. Weber, Q. Huo, and P. Rezamand. 2021. Relationship between serum

metabolites and milk fatty acid with periparturient diseases in Pacific Northwest dairy farms. J.

Dairy Sci. Vol. 104, Suppl. 1): 37 (abstr.).

Conference Proceedings

University of Minnesota

Fernandes, L., Celestino, M. L., Menta, P. R., Silva, T. H., Paiva, D., Ribeiro, T. L., Caixeta, L. S., Noyes, N. R., Machado, V. S. (2021). Risk factors associated with intramammary infections in primiparous dairy cows in organic herds. (J. Dairy Sci, vol. 104, Suppl. 1., pp. 215--216).

Mosca FP, Dean CJ, Heins BJ, Machado VS, Pinedo PJ, Noyes NR, Caixeta LR. Description of Staphylococcus sp. intramammary infections in early lactation heifers on organic dairy farms. American Dairy Science Association. Virtual, July 2021. Abstract 399.

Mosca FP, Dean CJ, Ray T, Doster E, Fernandes L, Antunes A, Sharpe K, Feijoo V, Baumann C, Wehri T, Heins BJ, Pinedo PJ, Machado VS, Caixeta LR, Noyes NR. Associations Between Intramammary Infections Caused by Non-aureus Staphylococci, Streptococcus spp. and Streptococcus-like organisms and Staphylococcus aureus in Early Lactation in Organic Dairy Cows. National Mastitis Council Annual Meeting. Virtual, January 2021.

2020. Fernandes, I. Guimaraes, N. Noyes, L. Caixeta, V.S. Machado. Impact of subclinical mastitis detected in the first month of lactation on performance of organic dairy cows. Proceedings of the 101st Conference of Research Workers in Animal Diseases, Virtual, December 2020. Abstract 410.

Dean, C, Felipe Pena Mosca, Tui Ray, Bradley Heins, Pablo Pinedo, Vinicius Machado, Luciano Caixeta, Noelle Noyes. What Is the Microbiome, and Why Is It Important for Organic Livestock Production? 2020 ASA-CSSA-SSSA International Annual Meeting. Virtual, November 2020.

Pena Mosca, F., Dean, C.J., Caixeta, L.S., Ray, T., Heins, B., Machado, V.S., Pinedo, P.J., Noyes, N.R. “Description of the intramammary infections dynamics in early lactation heifer on organic dairy farms.” 54th Annual Conference of the American Association of Bovine Practitioners. Salt Lake City, UT, 2021

University of Tennessee

Zeng, X., J. Vidlund, B. Gillespie, L. Cao, H. Wang,  J. Lin  and O. Kerro Dego. 2021. Immunogenicity of the Novel Enterobactin Conjugate Vaccine in Dairy Cows, Conference of Research Workers On Animal Diseases (CRWAD), December 3 – 7, Chicago, IL. USA.

Gelalcha, B. D.,  D. B. Ensermu, S. Brown, B. E. Gillespie, G. E. Agga, D. H. D’Souza, C. Okafor, and O. Kerro Dego. Detection of Multidrug-Resistant Extended-Spectrum β-lactamase Producing E. coli from Bulk tank Milk Obtained from Dairy Cattle Farms, Conference of Research Workers On Animal Diseases (CRWAD), December 3 – 7, Chicago, IL. USA.

Vidlund, J., B. E. Gillespie, G. E. Agga and O. Kerro Dego. Efficacy of Novel Staphylococcal Surface Protein Vaccines Against Mastitis in Dairy Cattle, Conference of Research Workers On Animal Diseases (CRWAD), December 3 – 7, Chicago, IL. USA.

University of Vermont

Lalonde C, Kraft J, Choudhary RK, Bourne DE, Shangraw EM, McFadden TB, Zhao F-Q (2021) Intramammary lipopolysaccharide infusion alters the fatty acid profile in blood triacylglycerides and phospholipids of lactating dairy cows. ADSA Annul Meeting 2021.

Cornell University

Virtual MEX ConneXion Event “Selective Treatment of Mastitis Science into practice from a US perspective  22^th June 2021 Ghent University. Virtual

Workshop ”Prevention and control of environmental mastitis”. April 21^th 2021.  Meeting con liberi professionisti scandinavi. Virtual.

1^st Master Class in Udder Health. Onfarm Academy “Manejo de ambiente para controle da mastite” February 2^th 2021. Virtual.

Poster Presentations

University of Minnesota

Using Rapid Culture Systems to Guide Selective Treatment of Clinical Mastitis and at Dry-off. E. Royster, S. Godden, E. Royster and S. Rowe. 3 hrs Short Course #1 at 60^th Annu Meeting of the National Mastitis Council. Jan 20, 2021. (virtual)

Decision support tools for selective dry cow therapy. Rowe, S.M., A. Vasquez, S.M. Godden, D.V. Nydam. Proc. 60^th Annu. Conf National Mastitis Council. Jan. 25-28, 2021 (virtual).   

3 Bs of Udder Health: Bedding, Bacteria Load and Better Management. S. Godden, F. Pena Mosca, E. Royster, S. Rowe, J. Timmeran, B. Crooker. National Mastitis Council Webinar. Nov. 19, 2020 (virtual).

3 B’s of Udder Health: Bedding, Bacterial Load and Better Management. IFRAMIX CCPA Czech Producer Webinar. 1.5 hr. Mar. 18, 2021 (virtual)

Is selective dry cow therapy an opportunity for your herd?  Carver County Dairy Expo. Feb. 15, 2021 (virtual)

Relationship between recycled manure solids processing methods, bedding bacteria counts and udder health. Carver County Dairy Expo. Feb. 15, 2021. (virtual)