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Participants responded to the Saint George’s Respiratory Questionnaire (SGRQ) at Baseline, Week 2, 200 and Week 4 to evaluate the supplement’s effect on quality of life as measured through symptoms, 201 impact, and total score (Supplemental Table 5). Across total score and the subcategories of 202 symptoms, activity, and impact there was no significant change from baseline to Week 4 in either the 203 healthy or asthmatic participants (Table 4). However, impact scores in asthmatic participants trended 204 towards significance ( P =0.065). Of the participants that smoke, have asthma, or have asthma and 205 smoke, 36% noted an improvement in their overall health, 43% noted less frequent coughing, 43% 206 noted fewer instances of feeling short of breath, and 29% noted fewer cough or breathing-related 207 sleep disturbances. 208

3.4 Majority of participants would recommend probiotic and herbal blend 209

In an end of trial survey, 100% of participants who smoke would recommend the probiotic and 210 herbal blend to friends and family. 90% of asthmatic participants and 82% of healthy participants 211 would recommend the same. 212

3.5 Probiotic blend improves serum short chain fatty acid levels in asthmatic subjects 213

Serum was analyzed for changes in short chain fatty acid (SCFA) levels between baseline and Week 214 4. Participants with asthma showed significant changes in serum SCFA levels. Propionic acid 215 increased across all asthmatics (Figure 2A), propionic acid and isovaleric acid increased significantly 216 in asthmatic non-smokers (Figure 2B), and acetic acid and butyric acid significantly increased in 217 subjects with asthma who smoked (Figure 2C). Across all groups, isobutyric acid and valeric acid did 218 not show a significant change, and hexanoic acid levels were too low for detection. 219

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3.6

Gut microbiome profile differs in asthmatic subjects

To examine the effects of the probiotic on the intestinal microbiome for potentially adverse signals, 221 we collected stool samples at baseline and Week 4 and analyzed for participants’ gut microbiome 222 signatures. The overall alpha (intra-sample) and beta (inter-sample) diversity was not significantly 223 altered between baseline and after 4 weeks of probiotic administration (Shannon Diversity, P =0.644, 224 T-test; Chao1 Richness Index, P =0.665, T-test, Figure 3A. P=1, R2=0.007, PERMANOVA; 225 P=0.279, PERMDISP, Figure 3B), with only modest differences in colonization at the genus level 226 (Supplemental Table 6). 227 However, the patient’s asthma status did associate with differences in alpha and beta diversity 228 (Chao1, P =0.033 Figure 3C, and P =0.017, R2 = 0.0464, PERMANOVA; P=0.52805, PERMDISP, 229 Figure 3D). The most prominent differences at the genus level were higher relative abundance of E. 230 coli (Log 2 FC 26.6), Bacteroidetes dorei (Log 2 FC 24.2), and B. ovatus (Log 2 FC 21.7) (Supplemental 231 Table 7). 232

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3.7

Probiotic Lactobacillus strains detected in stool

We performed qPCR using primers validated for strain-specificity. Most samples were not colonized 234 with L. plantarum at baseline (19%), but this signature was augmented in most samples by 235 administration of the probiotic and herbal blend (81%, chi 2 , 12.5, P =0.0004). L. acidophilus 236 colonization was more common at baseline (37%) and was detected in most of the post-exposure 237 samples (62%, chi 2 2, P =0.157). Similarly, L. rhamnosus was detected in most individuals before and 238 after probiotic administration (62% versus 81%, chi 2 , 1.39, P =0.238). 239

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This is a provisional file, not the final typeset article

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