Caenorhabditis elegans naturally thrives in a soil environment where they feed on bacteria and are in constant association with a diverse range of microbes (Barker et al. 1994). C. elegans egg laying is delayed or reduced when animals are infected with Burkholderia pseudomallei, Burkholderia thailandensis, Staphylococcus aureus, and Serratia marcescens (Mallo et al. 2002; OQuinn et al. 2002; Irazoqui et al. 2010). These changes in egg laying may be a protective response to pathogenic bacteria. Mutants of TGF-β-like DBL-1 signaling pathway also display reduced brood size (Luo et al. 2009; Roberts et al. 2010). While the peak of egg-laying activity seen in normal animals between days two and four is depressed in
dbl-1 pathway mutants, the reproductive span of these
dbl-1 pathway mutants is increased to up to 13 days (Luo et al. 2009). To determine if the egg-laying observed during infection is DBL-1 pathway-dependent, we tested the effect of the DBL-1 signaling pathway on egg laying when C. elegans were fed on representative Gram-negative (S. marcescens) and Gram-positive bacteria (Staphylococcus epidermidis). Similar to previously published reports, we found that loss of DBL-1 pathway signaling decreases brood size and increases reproductive span in normal laboratory conditions (E. coli strain OP50 and 20C incubation) (Figure 1A and B) (Luo et al. 2009; Roberts et al. 2010). Here, we report three new results. First, brood size reductions caused by infection and by loss of DBL-1 signaling are independent (Figure 1A). Wild-type and
dbl-1(
nk3) animals both significantly decrease their brood size when grown on S. marcescens (p= 0.005 and p< 0.001, respectively).
dbl-1 mutant animals laid even fewer eggs than the wild-type animals on S. marcescens, suggesting that the reduced brood size phenotype is independently affected by both S. marcescens exposure and by loss of DBL-1 (p< 0.001). While the decrease in brood size of wild-type animals on S. epidermidis was not significant (p= 0.115), the decreased brood size of
dbl-1(
nk3) animals was significant on this pathogenic bacterial strain (p= 0.045). Indeed, the decreases in brood size upon infection with either S. marcescens or S. epidermidis in both wild-type and
dbl-1(
nk3) populations are similar (p= 0.57), suggesting the pathogenic bacteria affect brood size independent of DBL-1. Because
dbl-1(
nk3) populations display a further reduced brood size upon infection by either pathogen compared to the wild type, the negative effects of pathogen exposure and loss of DBL-1 signaling on brood size appear to be additive (p< 0.05). Second, while the wild-type population on S. marcescens survived until all animals ceased laying eggs, all
dbl-1(
nk3) animals died on S. marcescens by Day 5. These results explain why the extended reproductive span normally seen in
dbl-1(
nk3) populations was not observed on S. marcescens. These results also support previous reports of decreased viability of
dbl-1 mutant animals on another variety of S. marcescens, Db11 (Mallo et al. 2002). Third, S. epidermidis affects egg-laying patterns similar to loss of
dbl-1 function. Initially, both wild-type and
dbl-1(
nk3) strains on S. epidermidis have reduced eggs laid in the first four days compared to strains grown on the E. coli control. Loss of DBL-1 further reduced the number of eggs laid during each of these days, suggesting that this phenotype is independently affected by both S. epidermidis exposure and by loss of DBL-1 (p= 0.004). Then, both wild-type and
dbl-1(
nk3) strains on S. epidermidis have similar extended reproductive spans, extending to at least day 13 (one tenacious wild-type hermaphrodite laid embryos until day 15). This S. epidermidis-induced reproductive span extension appears to be independent of DBL-1 signaling, because the numbers of eggs laid by both wild-type and
dbl-1(
nk3) populations between days 5 and 16 were similar at these time points (p= 0.509).