Ten best practices for taking experiential learning online.

Like many institutions around the world, the COVID-19 pandemic prompted us to shift our summer 2020 in-person undergraduate experiential learning program to a remote, virtual format. Here, we present our observations, summarized in 10 best practices, for moving a STEM-focused research experience for undergraduates, experiential learning program or research-based course online. We will also discuss how our program was originally designed and implemented, and how we adapted our activities to deliver an at-home research experience that maintained student engagement, mentorship, and a shared sense of community.

Exploring Rain as Source of Biological Control Agents for Fire Blight on Apple.

Poor survival on plants can limit the efficacy of Biological Control Agents (BCAs) in the field. Yet bacteria survive in the atmosphere, despite their exposure to high solar radiation and extreme temperatures. If conditions in the atmosphere are similar to, or more extreme than, the environmental conditions on the plant surface, then precipitation may serve as a reservoir of robust BCAs. To test this hypothesis, two hundred and fifty-four rain-borne isolates were screened for in vitro inhibition of Erwinia amylovora, the causal agent of fire blight, as well as of other plant pathogenic bacteria, fungi and oomycetes. Two isolates showed strong activity against E. amylovora and other plant pathogenic bacteria, while other isolates showed activity against fungal and oomycete pathogens. Survival assays suggested that the two isolates that inhibited E. amylovora were able to survive on apple blossoms and branches similarly to E. amylovora. Pathogen population size and associated fire blight symptoms were significantly reduced when detached apple blossoms were treated with the two isolates before pathogen inoculation, however, disease reduction on attached blossoms within an orchard was inconsistent. Using whole genome sequencing, the isolates were identified as Pantoea agglomerans and P. ananatis, respectively. A UV-mutagenesis screen pointed to a phenazine antibiotic D-alanylgriseoluteic acid synthesis gene cluster as being at the base of the antimicrobial activity of the P. agglomerans isolate. Our work reveals the potential of precipitation as an under-explored source of BCAs, whole genome sequencing as an effective approach to precisely identify BCAs, and UV-mutagenesis as a technically simple screen to investigate the genetic basis of BCAs. More field trials are needed to determine the efficacy of the identified BCAs in fire blight control.

Drip Line Flushing with Chlorine May Not Be Effective in Reducing Bacterial Loads in Irrigation Water Distribution Systems.

Irrigation water distribution systems are used to supply water to produce crops, but the system may also provide a protected environment for the growth of human pathogens present in irrigation water. In this study, the effects of drip tape installation depth and sanitization on the microbial quality of irrigation groundwater were evaluated. Drip tape lines were installed on the soil surface or 5 or 10 cm below the soil surface. Water samples were collected from the irrigation source and the end of each drip line every 2 weeks over an 11-week period, and the levels of Escherichia coli, total coliforms, aerobic mesophilic bacteria, and enterococci were quantified. Half of the lines installed at each depth were flushed with sodium hypochlorite for 1 h during week 6 to achieve a residual of 10 ppm at the end of the line. There was a statistically significant (P = 0.01) effect of drip tape installation depth and sanitizer application on the recovery of E. coli, with increased levels measured at the 5-cm depth and in nonsanitized lines, although the levels were at the limit of detection, potentially confounding the results. There was no significant effect of drip tape depth on total coliforms, aerobic mesophiles, or enterococci. In contrast, a statistically significant increase (P < 0.01) in the recovery of total coliforms was recorded from the ends of lines that received chlorine. This may be indicative of shedding of cells owing to degradation of biofilms that formed on the inner walls of the lines. These findings emphasize the need to better understand conditions that may lead to corrosion and increases in bacterial loads inside drip lines during flushing. Recommendations to growers should suggest collecting groundwater samples for testing at the end of drip lines rather than at the source. Guidelines on flushing drip lines with chlorine may need to include water pH monitoring, a parameter that influences the corrosive properties of chlorine.

Effects of Cover Crop Species and Season on Population Dynamics of Escherichia coli and Listeria innocua in Soil.

Cover crops provide several ecosystem services, but their impact on enteric bacterial survival remains unexplored. The influence of cover cropping on foodborne pathogen indicator bacteria was assessed in five cover crop/green manure systems: cereal rye, hairy vetch, crimson clover, hairy vetch-rye and crimson clover-rye mixtures, and bare ground. Cover crop plots were inoculated with Escherichia coli and Listeria innocua in the fall of 2013 and 2014 and tilled into the soil in the spring to form green manure. Soil samples were collected and the bacteria enumerated. Time was a factor for all bacterial populations studied in all fields (P < 0.001). E. coli levels declined when soil temperatures dipped to <5°C and were detected only sporadically the following spring. L. innocua diminished somewhat but persisted, independently of season. In an organic field, the cover crop was a factor for E. coli in year 1 (P = 0.004) and for L. innocua in year 2 (P = 0.011). In year 1, E. coli levels were highest in the rye and hairy vetch-rye plots. In year 2, L. innocua levels were higher in hairy vetch-rye (P = 0.01) and hairy vetch (P = 0.03) plots than in the rye plot. Bacterial populations grew (P < 0.05) or remained the same 4 weeks after green manure incorporation, although initial reductions in L. innocua numbers were observed after tilling (P < 0.05). Green manure type was a factor only for L. innocua abundance in a transitional field (P < 0.05). Overall, the impacts of cover crops/green manures on bacterial population dynamics in soil varied, being influenced by bacterial species, time from inoculation, soil temperature, rainfall, and tillage; this reveals the need for long-term studies.

Evaluation of Rotational Biopesticide Programs for Disease Management in Organic Cucurbit Production.

Downy mildew (Pseudoperonospora cubensis) and powdery mildew (Podosphaera xanthii) are two of the most economically important and widespread cucurbit diseases. Disease management relies primarily on fungicide use, but frequent fungicide applications can lead to the development of resistant pathogen populations. In addition, more vegetables are being produced with organic practices, which prohibit the use of many fungicides. Incorporating biorational products into a disease management program may help mitigate the risk of fungicide resistance development while being compatible with organic production. Field trials were conducted for two years on organically managed land in Maryland with cucumber, muskmelon, pumpkin, and butternut squash to evaluate the efficacy of four biorational products (i.e., Actinovate AG, OxiDate, Regalia, and Serenade Soil) when applied in a rotational program with copper against foliar cucurbit diseases. Generally, all biorational treatments resulted in significantly lower downy and powdery mildew severity compared with the nontreated plants, but the level of disease management was not significantly different than that provided by copper alone. However, Actinovate AG, OxiDate, and Serenade Soil each improved disease management on at least one crop, as compared with copper alone. Rotational programs with biopesticides are a viable disease management option for organic production of field-grown cucurbits in Maryland.

The growing season, but not the farming system, is a food safety risk determinant for leafy greens in the mid-Atlantic region of the United States.

Small- and medium-size farms in the mid-Atlantic region of the United States use varied agricultural practices to produce leafy greens during spring and fall, but the impact of preharvest practices on food safety risk remains unclear. To assess farm-level risk factors, bacterial indicators, Salmonella enterica, and Shiga toxin-producing Escherichia coli (STEC) from 32 organic and conventional farms were analyzed. A total of 577 leafy greens, irrigation water, compost, field soil, and pond sediment samples were collected. Salmonella was recovered from 2.2% of leafy greens (n = 369) and 7.7% of sediment (n = 13) samples. There was an association between Salmonella recovery and growing season (fall versus spring) (P = 0.006) but not farming system (organic or conventional) (P = 0.920) or region (P = 0.991). No STEC was isolated. In all, 10% of samples were positive for E. coli: 6% of leafy greens, 18% of irrigation water, 10% of soil, 38% of sediment, and 27% of compost samples. Farming system was not a significant factor for levels of E. coli or aerobic mesophiles on leafy greens but was a significant factor for total coliforms (TC) (P < 0.001), with higher counts from organic farm samples. Growing season was a factor for aerobic mesophiles on leafy greens (P = 0.004), with higher levels in fall than in spring. Water source was a factor for all indicator bacteria (P < 0.001), and end-of-line groundwater had marginally higher TC counts than source samples (P = 0.059). Overall, the data suggest that seasonal events, weather conditions, and proximity of compost piles might be important factors contributing to microbial contamination on farms growing leafy greens.

Assessment of region, farming system, irrigation source and sampling time as food safety risk factors for tomatoes.

In the mid-Atlantic region of the United States, small- and medium-sized farmers use varied farm management methods and water sources to produce tomatoes. It is unclear whether these practices affect the food safety risk for tomatoes. This study was conducted to determine the prevalence, and assess risk factors for Salmonella enterica, Shiga toxin-producing Escherichia coli (STEC) and bacterial indicators in pre-harvest tomatoes and their production areas. A total of 24 organic and conventional, small- to medium-sized farms were sampled for six weeks in Maryland (MD), Delaware (DE) and New Jersey (NJ) between July and September 2012, and analyzed for indicator bacteria, Salmonella and STEC. A total of 422 samples--tomato fruit, irrigation water, compost, field soil and pond sediment samples--were collected, 259 of which were tomato samples. A low level of Salmonella-specific invA and Shiga toxin genes (stx1 or stx2) were detected, but no Salmonella or STEC isolates were recovered. Of the 422 samples analyzed, 9.5% were positive for generic E. coli, found in 5.4% (n=259) of tomato fruits, 22.5% (n=102) of irrigation water, 8.9% (n=45) of soil, 3/9 of pond sediment and 0/7 of compost samples. For tomato fruit, farming system (organic versus conventional) was not a significant factor for levels of indicator bacteria. However, the total number of organic tomato samples positive for generic E. coli (1.6%; 2/129) was significantly lower than for conventional tomatoes (6.9% (9/130); (χ(2) (1)=4.60, p=0.032)). Region was a significant factor for levels of Total Coliforms (TC) (p=0.046), although differences were marginal, with western MD having the highest TC counts (2.6 log CFU/g) and NJ having the lowest (2.0 log CFU/g). Tomatoes touching the ground or plastic mulch harbored significantly higher levels of TC compared to vine tomatoes, signaling a potential risk factor. Source of irrigation water was a significant factor for all indicator bacteria (p<0.0001), and groundwater had lower bacterial levels than surface water. End of line surface water samples were not significantly different from source water samples, but end of line groundwater samples had significantly higher bacterial counts than source (p<0.0001), suggesting that Good Agricultural Practices that focus on irrigation line maintenance might be beneficial. In general, local effects other than cropping practices, including topography, land use and adjacent industries, might be important factors contributing to microbiological inputs on small- and medium-sized farms in the mid-Atlantic region.