Relationship Between Facility Type and Bulk Tank Milk Bacteriology, Udder Health, Udder Hygiene, and Milk Production on Vermont Organic Dairy Farms.

The primary objective of this cross-sectional observational study was to determine whether bulk tank milk quality, udder health, udder hygiene and milk production outcomes were associated with facility type on organic dairies. A secondary objective was to identify other management-related risk factors associated with bulk tank milk quality, udder health, udder hygiene, and milk production on organic dairy herds in Vermont. We aimed to enroll 40 farms, to compare herds using the 2 most common systems (freestalls, tiestalls) for housing organic dairy cattle in the state with those using a bedded pack during the non-grazing season (typically November-May). Two general styles of bedded packs were observed: cultivated bedded packs and untilled deep bedded packs. Due to the limited number of herds using bedded packs to house lactating dairy cattle in Vermont, we combined untilled and cultivated bedded packs to describe udder hygiene, milk quality, and udder health on these loose-housing systems deeply-bedded with organic material. The study was completed on 21 farms (5 bedded packs, 6 freestalls, 10 tiestalls) before interruption due to the COVID-19 pandemic. Data captured from Dairy Herd Improvement Association records from the test closest to the date of the farm visit included average somatic cell score (SCS), standardized 150-d milk (pounds), % cows with current high SCS (SCS ≥ 4.0), % cows with newly elevated SCS (previous SCS < 4.0 to current ≥ 4.0), and % cows with chronically elevated SCS (SCS ≥ 4.0 last 2 tests). Multivariable linear regression models were used to describe outcomes by facility type, but suffered from limited statistical power due to small group sample sizes. Unconditional comparisons failed to find statistically significant differences between farms grouped by facility type in metrics captured from Dairy Herd Improvement Association test data, bulk tank milk somatic cell count (BTSCC) and aerobic culture data, or udder hygiene scores. A secondary analysis was conducted using univariate linear regression to identify associations between herd management factors and outcomes for all 21 farms combined. Although not all differences found were statistically significant in this secondary analysis combining all farms, numeric differences that may be biologically important are reported showing farms with deeper bedding had a lower BTSCC, lower newly elevated SCS, lower chronically elevated SCS, lower elevated current SCS, lower average SCS, and better udder hygiene metrics. Farms with lower mean udder hygiene scores had numerically lower chronically elevated SCS, lower elevated current SCS, and lower average SCS. We could not reject the null hypothesis that milk quality and udder health outcomes did not differ by facility type, and this does not preclude the existence of biological differences in these outcomes between facility types. The current study provides insight on factors affecting bulk tank milk quality, udder health and hygiene measures on organic dairy farms in Vermont. Bedded packs may be a viable option for confinement housing during the winter non-grazing season for pasture-based herds interested in a loose-housing system in the Northeastern US, but more research such as longitudinal studies with a larger sample size is needed to test this hypothesis.

Anaerobic Soil Disinfestation and Vermicompost to Manage Bottom Rot in Organic Lettuce.

Rhizoctonia solani Kühn (teleomorph: Thanatephorus cucumeris [Frank] Donk) is an aggressive soilborne pathogen with a wide host range that survives saprophytically between crops, presenting a challenge for organic vegetable farmers who lack effective management tools. A 2-year field experiment was conducted at two organic farms to compare anaerobic soil disinfestation (ASD) and worm-cured compost (vermicompost) to manage bottom rot caused by R. solani subspecies AG1-IB in field-grown organic lettuce (Lactuca sativa). At each farm, four replicate plots of seven treatments were arranged in a randomized complete block design. Randomization was restricted by grouping treatments to evaluate ASD, and treatments to evaluate vermicompost in starter plugs. ASD experiment treatments were three different ASD carbon sources that are commonly used and widely available to local farmers in Vermont: compost, cover crop residues, and poultry manure fertilizer, as well as a tarped control. Vermicompost experimental treatments were vermicompost compared with two types of controls: a commercial biocontrol product (RootShield PLUS + G), and unamended (untarped control). This study demonstrated that the ASD method is achievable in a field setting on Vermont farms. However, neither ASD nor vermicompost produced significant disease suppression or resulted in higher marketable yields than standard growing practices. Given the laborious nature of ASD, it is likely more appropriate in a greenhouse setting with high-value crops that could especially benefit from being grown in plastic tarped beds (e.g., tomatoes and strawberries). This study is the first known attempt of field-implemented ASD for soil pathogen control in the northeastern United States.

Rhizoctonia solani AG1-IB, AG1-IC, and AG4-HGII cause bottom rot of field lettuce in Vermont.

Members of Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) species complex cause bottom rot on lettuce (Latuca sativa) and yield losses up to 70% (Subbarao et al. 2017). Severe symptoms include necrosis, stem rot, and/or discoloration especially on the leaf midrib. In Vermont, vegetable farms are small (0.5-30 acres) and grow lettuce concurrently with other vegetable crops in the same field but the AG(s) that causes the disease in Vermont has not been determined. Isolates (n = 157) were collected from 31 fields with reported history of bottom rot between July 10 and October 8, 2019, across Addison, Caledonia, Chittenden, Franklin, Lamoille, and Orleans counties. Isolates were collected from lettuce tissue or potato (Solanum tuberosum), a common rotation crop, or uncropped soil baited using radish (Raphanus sativus). Pieces of tissue (5-10 mm) were cut from the leading margin of lesions, surface disinfested with 0.1% NaClO for 1 min followed by 2 rinses with sterile water, blotted dry, and plated onto acidified 2% water agar (0.085% lactic acid, pH 4.8). After incubation for 48 to 72 h, mycelia resembling Rhizoctonia were examined for morphological characteristics including hyphal branching at ca. 90o angles, a septum near the branching point, multiple nuclei per cell, and lack of both clamp connections and conidia (Sneh et al. 1991). Colonies were white to dark brown, and some produced small sclerotia. Koch's postulates were performed by inoculating nine 8-week-old (9 leaf pairs) romaine lettuce plants (Johnny's Seeds, Winslow, ME, cv. Monte Carlo) per isolate. Isolates were grown on 2% potato dextrose agar for 1 week, from which a 5-mm agar plug was placed on the adaxial leaf surface at the base of a petiole. Plants were enclosed in a plastic bag to maintain high humidity and grown under a 16-hour photoperiod at 24 °C. Disease severity was rated 4 days after inoculation (0: healthy, 1: isolated lesions, 2: lesions across multiple petioles, and 3: systemic disease). Putative AG were determined by Sanger sequencing of the internal transcribed spacer (ITS) region using the ITS1F and ITS4B primer pair (758 bp) (Gardes and Bruns 1993). Contigs were assembled using CAP3 software (Huang and Madan 1999). Taxonomy was assigned to each OTU via the NCBI BLASTn database with criteria as 0.0 E and nucleotide match of at least 97%. Of the 10 isolates sequenced with sufficient coverage (735 to 784 bp alignment length) and definitive resolution (96.7 to 99.9% identity), 5 were putative AG 1-IB (Genbank Accession HG934430.1), 2 AG 1-IC (Genbank Accession AF354058.1), 2 AG 3 (Genbank Accession AF354064.1), and 1 AG 4-HGII (Genbank Accession AF354074.1). Fasta files and metadata are archived at 10.6084/m9.figshare.20301324, 10.6084/m9.figshare.20301375. Putative AG 1-IB was highly virulent on lettuce plants whether it originated from potato (mean 2.6) or lettuce (mean 1.3 to 3). AG 4-HGII and AG 1-IC isolated from lettuce and radish, respectively, were moderately severe (mean 1.4 to 2.2) on lettuce with identical symptoms. The two potato isolates (AG3) were not pathogenic on lettuce. Similarly, higher incidence of AG 1-IB is reported on lettuce in Quebec (Wallon et al. 2021), Ohio (Herr 1993), and Germany (Grosch et al. 2004). Because AG vary in their host range (Sneh et al. 1991), knowing the AG will inform management decisions such as crop rotation and weed control. This is the first report of the causal agent of bottom rot of lettuce or any AG of R. solani in Vermont.

Organic Amendments Alter Soil Hydrology and Belowground Microbiome of Tomato (Solanum lycopersicum).

Manure-derived organic amendments are a cost-effective tool that provide many potential benefits to plant and soil health including fertility, water retention, and disease suppression. A greenhouse experiment was conducted to evaluate how dairy manure compost (DMC), dairy manure compost-derived vermicompost (VC), and dehydrated poultry manure pellets (PP) impact the tripartite relationship among plant growth, soil physiochemical properties, and microbial community composition. Of tomato plants with manure-derived fertilizers amendments, only VC led to vigorous growth through the duration of the experiment, whereas DMC had mixed impacts on plant growth and PP was detrimental. Organic amendments increased soil porosity and soil water holding capacity, but delayed plant maturation and decreased plant biomass. Composition of bacterial communities were affected more by organic amendment than fungal communities in all microhabitats. Composition of communities outside roots (bulk soil, rhizosphere, rhizoplane) contrasted those within roots (endosphere). Distinct microbial communities were detected for each treatment, with an abundance of Massilia, Chryseolinea, Scedosporium, and Acinetobacter distinguishing the control, vermicompost, dairy manure compost, and dehydrated poultry manure pellet treatments, respectively. This study suggests that plant growth is affected by the application of organic amendments not only because of the soil microbial communities introduced, but also due to a synergistic effect on the physical soil environment. Furthermore, there is a strong interaction between root growth and the spatial heterogeneity of soil and root-associated microbial communities.

Design and implementation of a survey quantifying winter housing and bedding types used on Vermont organic dairy farms.

We conducted a descriptive observational study to quantify the frequency and diversity of winter housing and bedding types used by organic dairy farmers in Vermont. This report describes the survey methods, results, successes, limitations, and lessons learned from administering the survey. Beginning in December 2018, a short questionnaire was administered by web, mail, and telephone to a source population defined as all producers of organic dairy cow milk in Vermont (n = 177) listed in the United States Department of Agriculture Organic Integrity database. Our approach yielded an 82% (n = 145) response from certified organic farms producing cow milk in Vermont at the time of the survey. The 3 most common housing and bedding material combinations used by respondents were tiestall housing with wood (sawdust or shavings) bedding materials (45%), freestall housing with wood bedding materials (14%), and freestall housing with sand bedding (12%). Fifteen percent of respondents reported using more than one type of facility for winter housing of lactating cattle. The median number of lactating cows on farms among respondents was 59.5 (range: 2-400), and the odds of using more than one type of facility to house lactating cows increased positively with the number of lactating cows reported for a herd. Breed distribution was similar across the housing and bedding type categories. An association between frequency of individual cow milk somatic cell count testing and housing type was identified; respondents using freestall sand facilities tested less frequently than respondents using tiestalls with wood bedding. Although the questionnaire length limited the amount of information gathered, the response proportion was exceptional, and overall our survey results provide valuable insight on Vermont organic dairy housing and bedding practices that should inform future extension and outreach efforts for this sector of the dairy industry.

Mammary microbiome of lactating organic dairy cows varies by time, tissue site, and infection status.

Infections of the cow udder leading to mastitis and reducing milk quality are a critical challenge facing all dairy farmers. Mastitis may be linked to the ecological disruption of an endogenous mammary microbial community, suggesting an ecosystems approach to management and prevention of this disease. The teat end skin represents a first point of host contact with mastitis pathogens and may offer an opportunity for microbially mediated resistance to infection, yet we know little about the microbial community of teat end skin or its potential interaction with the microbial community of intramammary milk of organic dairy cattle. High-throughput sequencing of marker genes for bacterial and fungal communities was used to characterize the skin and milk microbiome of cows with both a healthy and infected gland (i.e., udder quarter) and to assess the sharing of microbial DNA between these tissue habitat sites. The mammary microbiome varied among cows, through time, and between skin and milk. Microbiomes of milk from healthy and infected quarters reflected a diverse group of microbial DNA sequences, though milk had far fewer operational taxonomic units (OTUs) than skin. Milk microbiomes of infected quarters were generally more variable than healthy quarters and were frequently dominated by a single OTU; teat end skin microbiomes were relatively similar between healthy and infected quarters. Commonly occurring genera that were shared between skin and milk of infected glands included Staphylococcus spp. bacteria and Debaryomyces spp. fungi. Commonly occurring genera that were shared between skin and milk of healthy glands included bacteria SMB53 (Clostridiaceae) and Penicillium spp. fungi. Results support an ecological interpretation of the mammary gland and the notion that mastitis can be described as a dysbiosis, an imbalance of the healthy mammary gland microbiome.