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.