Internal extracellular bacteria of Diaphorina citri Kuwayama (Hemiptera: Psyllidae), the Asian citrus psyllid.

The Asian Citrus Psyllid, Diaphorina citri Kuwayama (Hemiptera: Psyllidae), is an invasive insect pest that transmits Candidatus Liberibacter spp. This insect/pathogen system was first identified in North America in the early 2000's and has become the top threat to the citrus industry. Limited options for management of this problem exist; therefore, innovative pest management strategies are being developed. In this study, we describe the first step toward a paratransgenic approach (also referred to symbiotic control) for control of the insect vector or the pathogen. Culturable bacteria from the gut of Asian Citrus Psyllids were identified using standard culture techniques followed by sequencing of the cultured microorganisms. Further, 454 pyrosequencing of the gut was performed to audit bacterial presence in order to begin to identify any relationship between psyllid symbionts and C. Liberibacter spp.

Bacterial community composition of three candidate insect vectors of palm phytoplasma (Texas Phoenix Palm Decline and Lethal Yellowing).

Texas Phoenix Palm Decline (TPPD) and Lethal Yellowing are two phytoplasma-linked diseases in palms. The phytoplasma causing TPPD is thought to be transmitted by three putative planthopper vectors, Ormenaria rufifascia, Omolicna joi, and Haplaxius crudus. These insects have been morphologically and molecularly described, and have screened positive for Candidatus Phytoplasma palmae. Individuals from each species were subjected to 16S bacterial community sequencing using the Roche 454 platform, providing new information regarding the previously unexplored bacterial communities present in putative vectors.

Propagation of Homalodisca coagulata virus-01 via Homalodisca vitripennis cell culture.

The glassy-winged sharpshooter (Homalodisca vitripennis) is a highly vagile and polyphagous insect found throughout the southwestern United States. These insects are the predominant vectors of Xylella fastidiosa (X. fastidiosa), a xylem-limited bacterium that is the causal agent of Pierce's disease (PD) of grapevine. Pierce's disease is economically damaging; thus, H. vitripennis have become a target for pathogen management strategies. A dicistrovirus identified as Homalodisca coagulata virus-01 (HoCV-01) has been associated with an increased mortality in H. vitripennis populations. Because a host cell is required for HoCV-01 replication, cell culture provides a uniform environment for targeted replication that is logistically and economically valuable for biopesticide production. In this study, a system for large-scale propagation of H. vitripennis cells via tissue culture was developed, providing a viral replication mechanism. HoCV-01 was extracted from whole body insects and used to inoculate cultured H. vitripennis cells at varying levels. The culture medium was removed every 24 hr for 168 hr, RNA extracted and analyzed with qRT-PCR. Cells were stained with trypan blue and counted to quantify cell survivability using light microscopy. Whole virus particles were extracted up to 96 hr after infection, which was the time point determined to be before total cell culture collapse occurred. Cells were also subjected to fluorescent staining and viewed using confocal microscopy to investigate viral activity on F-actin attachment and nuclei integrity. The conclusion of this study is that H. vitripennis cells are capable of being cultured and used for mass production of HoCV-01 at a suitable level to allow production of a biopesticide.

Bacterial community survey of Solenopsis invicta Buren (red imported fire ant) colonies in the presence and absence of Solenopsis invicta virus (SINV).

Insect bacterial symbionts contribute to many essential biological functions of their hosts and can also influence host fecundity and fitness. The physiological contribution symbionts provide can aid in immune response and xenobiotic detoxification. Both of these immune factors can directly impact strategies aimed at managing insect populations. One biological control strategy that shows promise in insects is the use of single-stranded RNA viruses within the group Dicistroviridae. The Solenopsis invicta Virus (SINV; Dicistroviridae), a ssRNA virus, has been proposed as a potential biological control agent for the urban pest S. invicta Buren or red imported fire ant (RIFA). SINV has been shown to be prevalent in RIFA populations of Texas and Florida; however, mortality is associated with high viral load. In other insect microbe systems, presence of particular bacteria induced resistance against Dicistrovirus. If this type of relationship is present in the RIFA-SINV system, their bacterial community could reduce the effectiveness of SINV as a biological control system. The advantage of 454 pyro-sequencing is that it enables classification of unculturable bacteria. This study examines the bacterial community in brood, workers, and reproductive cast members from colonies with and without SINV infection. Manipulation of the bacterial community may alter virus infection and replication within the mid-gut. Understanding the differences in the microbial community of ant colonies may provide insights that will refine current efforts designing control strategies for this important urban pest.

Recreational boating traffic: a chronic source of anthropogenic noise in the Wilmington, North Carolina Intracoastal Waterway.

The majority of attention on the impact of anthropogenic noise on marine mammals has focused on low-frequency episodic activities. Persistent sources of mid-frequency noise pollution are less well studied. To address this data gap, the contribution of 25 physical, biological and anthropogenic factors to the ambient noise levels in the Wilmington, North Carolina Intracoastal Waterway were analyzed using a principal components analysis and least squares regression. The total number of recreational vessels passing through the waterway per hour is the factor that had the single greatest influence on environmental noise levels. During times of high boat traffic, anthropogenic noise is continuous rather than episodic, and occurs at frequencies that are biologically relevant to bottlenose dolphins. As a daily part of resident bottlenose dolphins' acoustic environment, recreational boating traffic may represent a chronic source of acoustic harassment.