Enterococcus faecalis is a Gram-positive bacterium that is frequently found in the periapical lesion of patients with apical periodontitis. Its biofilm formation in root canal is closely related to the development of refractory apical periodontitis by providing increased resistance to endodontic treatments.
Phage therapy has recently been considered as an efficient therapeutic strategy in controlling various periodontal pathogens. We previously demonstrated the bactericidal capacities of Enterococcus phage vB_EfaS_HEf13 (phage HEf13) against clinically-isolated E. faecalis strains. Here, we investigated whether phage HEf13 affects biofilm formation and pre-formed biofilm of clinically-isolated E.
faecalis, and its combinatory effect with endodontic treatments, including chlorhexidine (CHX) and penicillin. The phage HEf13 inhibited biofilm formation and disrupted pre-formed biofilms of E. faecalis in a dose- and time-dependent manner. Interestingly, phage HEf13 destroyed E. faecalis biofilm exopolysaccharide (EPS), which is known to be a major component of bacterial biofilm. Furthermore, combined treatment of phage HEf13 with CHX or penicillin more potently inhibited biofilm formation and disrupted pre-formed biofilm than either treatment alone. Confocal laser scanning microscopic examination demonstrated that these additive effects of the combination treatments on disruption of pre-formed biofilm are mediated by relatively enhanced reduction in thickness distribution and biomass of biofilm. Collectively, our results suggest that the effect of phage HEf13 on E. faecalis biofilm is mediated by its EPS-degrading property, and its combination with endodontic treatments more potently suppresses E. faecalis biofilm, implying that phage HEf13 has potential to be used as a combination therapy against E. faecalis infections.
Reactive oxygen species induce DNA strand breaks and DNA oxidation. DNA oxidation leads to DNA mismatches, resulting
in mutations in the genome if not properly repaired. Homologous recombination (HR) and non-homologous end-joining
(NHEJ) are required for DNA strand breaks, whereas the base excision repair system mainly repairs oxidized DNAs, such as
8-oxoguanine and thymine glycol, by cleaving the glycosidic bond, inserting correct nucleotides, and sealing the gap. Our
previous studies revealed that the Rad53-Bdr1 pathway mainly controls DNA strand breaks through the regulation of HRand
NHEJ-related genes. However, the functional roles of genes involved in the base excision repair system remain elusive
in Cryptococcus neoformans. In the present study, we identified OGG1 and NTG1 genes in the base excision repair system
of C. neoformans, which are involved in DNA oxidation repair. The expression of OGG1 was induced in a Hog1-dependent
manner under oxidative stress. On the other hand, the expression of NTG1 was strongly induced by DNA damage stress in a
Rad53-independent manner. We demonstrated that the deletion of NTG1, but not OGG1, resulted in elevated susceptibility
to DNA damage agents and oxidative stress inducers. Notably, the ntg1Δ mutant showed growth defects upon antifungal
drug treatment. Although deletion of OGG1 or NTG1 did not increase mutation rates, the mutation profile of each ogg1Δ
and ntg1Δ mutant was different from that of the wild-type strain. Taken together, we found that DNA N-glycosylase Ntg1
is required for oxidative DNA damage stress and antifungal drug resistance in C. neoformans.
Dark septate endophytes (DSEs) are widely distributed and improve plant growth. DSEs secrete large amounts of enzymes
to mineralize insoluble phosphorus in soil and convert it into soluble phosphorus, promoting plant uptake of phosphorus.
However, the effects of DSEs with phosphate-solubilizing ability on host plants need further study. In this study, phosphorusdissolving
DSEs were screened for growth-promoting effects. We isolated, identified and characterized three DSE species
(Thozetella neonivea, Pezicula ericae and Hyaloscyphaceae sp.) showing phosphate-solubilizing ability. The impact of single,
dual or triple inoculation of DSEs on blueberry plant characteristics was studied. Their effects on colonization intensity,
seedling biomass, nutrients in plants and soil, and activities of plant resistance enzymes and soil enzymes were markedly
upregulated relative to the control (P < 0.05). The available phosphorus and acid phosphatase levels in different combinations
were significantly increased. These findings indicate that the application of the three DSEs may be valuable in facilitating
the cultivation of blueberry with a higher biomass and improved plant quality.
Streptococcus suis serotype 2 (SS2) is an important zoonotic pathogen that can infect humans in contact with infected pigs
or their byproducts. It can employ different types of genes to defend against oxidative stress and ensure its survival. The
thioredoxin (Trx) system is a key antioxidant system that contributes adversity adaptation and pathogenicity. SS2 has been
shown to encode putative thioredoxin genes, but the biological roles, coding sequence, and underlying mechanisms remains
uncharacterized. Here, we demonstrated that SSU05_0237-ORF, from a clinical SS2 strain, ZJ081101, encodes a protein
of 104 amino acids with a canonical CGPC active motif and an identity 70–85% similar to the thioredoxin A (TrxA) in
other microorganisms. Recombinant TrxA efficiently catalyzed the thiol-disulfide oxidoreduction of insulin. The deletion
of TrxA led to a significantly slow growth and markedly compromised tolerance of the pathogen to temperature stress,
as well as impaired adhesion ability to pig intestinal epithelial cells (IPEC-J2). However, it was not involved in H2O2
and
paraquat-induced oxidative stress. Compared with the wild-type strain, the ΔTrxA strain was more susceptible to killing by
macrophages through increasing NO production. Treatment with TrxA mutant strain also significantly attenuated cytotoxic
effects on RAW 264.7 cells by inhibiting inflammatory response and apoptosis. Knockdown of pentraxin 3 in RAW 264.7
cells was more vulnerable to phagocytic activity, and TrxA promoted SS2 survival in phagocytic cells depending on pentraxin
3 activity compared with the wild-type strain. Moreover, a co-inoculation experiment in mice revealed that TrxA mutant
strain is far more easily cleared from the body than the wild type strain in the period from 8–24 h, and exhibits significantly
attenuated oxidative stress and liver injury. In summary, we reveal the important role of TrxA in the pathogenesis of SS2.