The Biology of Casmara subagronoma (Lepidoptera: Oecophoridae), a Stem-Boring Moth of Rhodomyrtus tomentosa (Myrtaceae): Descriptions of the Previously Unknown Adult Female and Immature Stages, and Its Potential as a Biological Control Candidate.

Rhodomyrtus tomentosa is a perennial shrub native to Southeast Asia and is invasive in South Florida and Hawai'i, USA. During surveys of R. tomentosa in Hong Kong from 2013-2018 for potential biological control agents, we collected larvae of the stem borer, Casmara subagronoma. Larvae were shipped in stems to a USDA-ARS quarantine facility where they were reared and subjected to biology studies and preliminary host range examinations. Casmara subagronoma is the most recent Casmara species to be described from males collected in Vietnam and Indonesia. Because the original species description was based on only two male specimens, we also provide a detailed description of the female, egg, larva, and pupa. Finally, we conducted preliminary host range trials utilizing Myrtus communis, Myrcianthes fragrans, and Camellia sinensis. Casmara subagronoma emerged from M. fragrans, a Florida-native shrub, and larvae were able to survive in non-target stems for over a year (>400 days). Based on these findings and difficulty in rearing, we do not believe C. subagronoma is a suitable insect for biological control of R. tomentosa at this time, but may warrant further study. This investigation also illustrates the importance of host surveys for conservation and taxonomic purposes.

Bromate-induced Changes in p21 DNA Methylation and Histone Acetylation in Renal Cells.

Bromate (BrO3-) is a water disinfection byproduct (DBP) previously shown to induce nephrotoxicity in vitro and in vivo. We recently showed that inhibitors of DNA methyltransferase 5-aza-2'-deoxycytidine (5-Aza) and histone deacetylase trichostatin A (TSA) increased BrO3- nephrotoxicity whereas altering the expression of the cyclin-dependent kinase inhibitor p21. Human embryonic kidney cells (HEK293) and normal rat kidney (NRK) cells were sub-chronically exposed to BrO3- or epigenetic inhibitors for 18 days, followed by 9 days of withdrawal. DNA methylation was studied using a modification of bisulfite amplicon sequencing called targeted gene bisulfite sequencing. Basal promoter methylation in the human p21 promoter region was substantially lower than that of the rat DNA. Furthermore, 5-Aza decreased DNA methylation in HEK293 cells at the sis-inducible element at 3 distinct CpG sites located at 691, 855, and 895 bp upstream of transcription start site (TSS). 5-Aza also decreased methylation at the rat p21 promoter about 250 bp upstream of the p21 TSS. In contrast, sub-chronic BrO3- exposure failed to alter methylation in human or rat renal cells. BrO3- exposure altered histone acetylation in NRK cells at the p21 TSS, but not in HEK293 cells. Interestingly, changes in DNA methylation induced by 5-Aza persisted after its removal; however, TSA- and BrO3--induced histone hyperacetylation returned to basal levels after 3 days of withdrawal. These data demonstrate novel sites within the p21 gene that are epigenetically regulated and further show that significant differences exist in the epigenetic landscape between rat and human p21, especially with regards to toxicant-induced changes in histone acetylation.