Pennsylvania responded to the COVID-19 pandemic by closing schools and moving to online instruction in March 2020. We surveyed Pennsylvania school nurses ( N = 350) in May 2020 to assess the impact of COVID-19 on nurses’ concerns about returning to school and impact on practice. Data were analyzed using χ2 tests and regression analyses. Urban school nurses were more concerned about returning to the school building without a COVID-19 vaccine than rural nurses ( OR = 1.58, 95% CI [1.05, 2.38]). Nurses in urban locales were more likely to report being asked for guidance on COVID-19 ( OR = 1.69, 95% CI [1.06, 2.68]), modify communication practices ( OR = 2.33, 95% CI [1.42, 3.82]), and be “very/extremely concerned” about their safety ( OR = 2.16, 95% CI [1.35, 3.44]). Locale and student density are important factors to consider when resuming in-person instruction; however, schools should recognize school nurses for their vital role in health communication to assist in pandemic preparedness and response.

25Stable associations between insects and bacterial species are widespread in nature. This 26 is the case for many economically important insects, such as tsetse flies. Tsetse flies are 27 the vectors of Trypanosoma brucei, the etiological agent of African trypanosomiasis-a 28 zoonotic disease that incurs a high socioeconomic cost in endemic regions. Populations 29 of tsetse flies are often infected with the bacterium Sodalis glossinidius. Following 30 infection, S. glossinidius establishes a chronic, stable association characterized by vertical 31 (maternal) and horizontal (paternal) modes of transmission. Due to the stable nature of 32 this association, S. glossinidius has been long sought as a means for the implementation 33 of anti-Trypanosoma paratransgenesis in tsetse flies. However, the lack of tools for the 34 genetic modification of S. glossinidius has hindered progress in this area. Here we 35 establish that S. glossinidius is amenable to DNA uptake by conjugation. We show that 36 conjugation can be used as a DNA delivery method to conduct forward and reverse 37 genetic experiments in this bacterium. This study serves as an important step in the 38 development of genetic tools for S. glossinidius. The methods highlighted here should 39 guide the implementation of genetics for the study of the tsetse-Sodalis association and 40 the evaluation of S. glossinidius-based tsetse fly paratransgenesis strategies. 41 42 Importance 43 Tsetse flies are the insect vectors of T. brucei, the causative agent of African sleeping 44 sickness-a zoonotic disease that inflicts a substantial economic cost to a broad region 45 of sub-Saharan Africa. Notably, tsetse flies can be infected with the bacterium S. 46 glossinidius to establish an asymptomatic chronic infection. This infection can be 47 inherited by future generations of tsetse flies allowing S. glossinidius to spread and 48 persist within populations. To this effect, S. glossinidius has been considered as a 49 3 potential expression platform to create flies which reduce T. brucei stasis and lower 50overall parasite transmission to humans and animals. However, the efficient genetic 51 manipulation of S. glossinidius has remained a technical challenge due to its complex 52 growth requirements and uncharacterized physiology. Here we exploit a natural 53 mechanism of DNA transfer among bacteria and develop an efficient technique to 54 genetically manipulate S. glossinidius for future studies in reducing trypanosome 55 transmission. 56 57 Results 100

Stable associations between insects and bacterial species are widespread in nature. This is the case for many economically important insects, such as tsetse flies. Tsetse flies are the vectors of Trypanosoma brucei, the etiological agent of African trypanosomiasis—a zoonotic disease that incurs a high socioeconomic cost in regions of endemicity. Populations of tsetse flies are often infected with the bacterium Sodalis glossinidius. Following infection, S. glossinidius establishes a chronic, stable association characterized by vertical (maternal) and horizontal (paternal) modes of transmission. Due to the stable nature of this association, S. glossinidius has been long sought as a means for the implementation of anti-Trypanosoma paratransgenesis in tsetse flies. However, the lack of tools for the genetic modification of S. glossinidius has hindered progress in this area. Here, we establish that S. glossinidius is amenable to DNA uptake by conjugation. We show that conjugation can be used as a DNA delivery method to conduct forward and reverse genetic experiments in this bacterium. This study serves as an important step in the development of genetic tools for S. glossinidius. The methods highlighted here should guide the implementation of genetics for the study of the tsetse-Sodalis association and the evaluation of S. glossinidius-based tsetse fly paratransgenesis strategies. IMPORTANCE Tsetse flies are the insect vectors of T. brucei, the causative agent of African sleeping sickness—a zoonotic disease that inflicts a substantial economic cost on a broad region of sub-Saharan Africa. Notably, tsetse flies can be infected with the bacterium S. glossinidius to establish an asymptomatic chronic infection. This infection can be inherited by future generations of tsetse flies, allowing S. glossinidius to spread and persist within populations. To this effect, S. glossinidius has been considered a potential expression platform to create flies which reduce T. brucei stasis and lower overall parasite transmission to humans and animals. However, the efficient genetic manipulation of S. glossinidius has remained a technical challenge due to its complex growth requirements and uncharacterized physiology. Here, we exploit a natural mechanism of DNA transfer among bacteria and develop an efficient technique to genetically manipulate S. glossinidius for future studies in reducing trypanosome transmission.

BACKGROUND All 50 states have school‐entry immunization requirements, and many also allow exemptions based on medical and non‐medical reasons. School nurses are responsible for managing student immunization compliance based on state policies, but lack standardized resources and guidance. METHODS Pennsylvania school nurses (N = 21) participated in semi‐structured interviews regarding their strategies for communication and management of student immunization information, along with resources needed for practice improvement. Data were analyzed using descriptive content analysis. RESULTS Nurses reported similarities in timelines used for communication of immunization requirements, but differences in mechanisms used to secure and manage immunization records. Nurses reported a need for clarity regarding exclusions and exemption policy implementation and requested standardized resources and guidance for navigating immunization compliance. CONCLUSIONS A need exists for standardized processes that support immunization compliance. Furthermore, nurses highlighted a need for additional training and enhanced networks to develop creative strategies for promoting immunization uptake among families.

Bacteriophages (phages) are ubiquitous in nature. These viruses play a number of central roles in microbial ecology and evolution by, for instance, promoting horizontal gene transfer (HGT) among bacterial species. The ability of phages to mediate HGT through transduction has been widely exploited as an experimental tool for the genetic study of bacteria. As such, bacteriophage P1 represents a prototypical generalized transducing phage with a broad host range that has been extensively employed in the genetic manipulation of Escherichia coli and a number of other model bacterial species. Here we demonstrate that P1 is capable of infecting, lysogenizing, and promoting transduction in members of the bacterial genus Sodalis, including the maternally inherited insect endosymbiont Sodalis glossinidius. While establishing new tools for the genetic study of these bacterial species, our results suggest that P1 may be used to deliver DNA to many Gram-negative endosymbionts in their insect host, thereby circumventing a culturing requirement to genetically manipulate these organisms. IMPORTANCE A large number of economically important insects maintain intimate associations with maternally inherited endosymbiotic bacteria. Due to the inherent nature of these associations, insect endosymbionts cannot be usually isolated in pure culture or genetically manipulated. Here we use a broad-host-range bacteriophage to deliver exogenous DNA to an insect endosymbiont and a closely related free-living species. Our results suggest that broad-host-range bacteriophages can be used to genetically alter insect endosymbionts in their insect host and, as a result, bypass a culturing requirement to genetically alter these bacteria.

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BackgroundThe whole school, whole community, whole child (WSCC) model suggests wellness councils, ongoing review of wellness policy, and a plan for evaluating set objectives are some of the key features needed to support school wellness infrastructure. This study explored the relationship between implementation of these infrastructure features and overall school wellness environment assessment scores among a sampling of Pennsylvania schools.MethodsThe Healthy Champions program provides Pennsylvania schools an opportunity to self‐assess their wellness environments across several school wellness topics. Staff enrolled their school in the program by completing a self‐report electronic assessment. Enrollment data from the 2020/2021 program year were analyzed using the Kruskal‐Wallis test and linear fixed model to identify the impact of varied implementation levels across 3 wellness infrastructure activities. Interactions between these variables and overall assessment score were also analyzed.ResultsOf the 645 Pennsylvania schools enrolled and analyzed, we observed higher mean wellness environment assessment scores (∆ 0.74 95% CI 0.40‐1.07; p < 0.001) among schools that reported some frequency of all 3 wellness infrastructure activities, compared to schools that reported no frequency for the activities.ImplicationsSchools with existing policies and practices related to the 3 wellness infrastructure activities should consider the degree of implementation to best support overall wellness in their school setting. Additional research to explore implementation barriers and supports is needed.ConclusionsAnalyses indicated that overall wellness environment assessment scores are impacted by implementation thresholds for wellness council meeting frequency, revision of wellness policy, and review of student health promotion objectives.

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