Tracking sero-molecular trends of swine brucellosis in Hawai'i and the central Pacific.

Introduction: Brucellosis is a zoonotic disease of mammals caused by bacterial species of the Brucella genus. The reservoir for disease is typically mammals, with species of Brucella found infecting amphibians, bats, and marine mammals. Brucella spp. can pass directly to humans through contact with infected animals or their products. Brucella spp. can cause chronic debilitating infections in mammals, including humans, and is associated with spontaneous abortions in infected animals, causing reduced fecundity. In Hawai'i, terrestrial species that could harbor Brucella spp. include swine, cattle, horses, and axis deer among others. The numerous feral swine in Hawai'i are known to carry Brucella suis, with evidence supporting infections in cattle. Brucella suis also poses infection risk to humans, dogs, and potentially horses across the state. Methods: In this study, 3,274 feral swine serum samples collected from 5 of the 8 main islands over a 15-year span were analyzed for exposure to B. suis. Of the 558 watersheds in the state, 77 were sampled as part of this effort. Spatial analysis was used to identify watersheds of concern. MLVA and whole genome SNP analysis was used for molecular epidemiological analysis. Results: Statewide seropositivity rates were triple that of feral swine found in the conterminous United States. Smoothed positivity rates were highest on Maui, followed by O'ahu, and the island of Hawai'i. Island-by-island analysis found high brucellosis positivity levels associated with specific watersheds and agricultural areas. Local spatial autocorrelation identified hot spots on O'ahu and Hawai'i. MLVA analysis of available B. suis from Hawai'i found molecular epidemiological connections with B. suis found in French Polynesia and the mainland US while differing from those in Tonga, Western Polynesia. Strains from Hawai'i are phylogenetically closest to strains from the United States. MLVA and SNP analysis found B. suis strains from Hawai'i fell into the genetic group that contains biovar 1 B. suis. Discussion: This work identified islands and watersheds of high brucellosis seropositivity in feral swine of Hawai'i, highlighting the magnitude of the zoonotic risk. Introduction of strains in recent history is unlikely due to modern animal trade and disease control practices. Genomic analysis of strains in Hawai'i and the Pacific area can provide hidden historical and local clues to brucellosis epidemiology in the state.

Mutation of the toxR or mshA genes from Vibrio coralliilyticus strain OCN014 reduces infection of the coral Acropora cytherea.

Thermal stress increases the incidence of coral disease, which is predicted to become more common with climate change, even on pristine reefs such as those surrounding Palmyra Atoll in the Northern Line Islands that experience minimal anthropogenic stress. Here we describe a strain of Vibrio coralliilyticus, OCN014, which was isolated from Acropora cytherea during an outbreak of Acropora white syndrome (AWS), a tissue loss disease that infected 25% of the A. cytherea population at Palmyra Atoll in 2009. OCN014 recreated signs of disease in experimentally infected corals in a temperature-dependent manner. Genes in OCN014 with expression levels positively correlated with temperature were identified using a transposon-mediated genetic screen. Mutant strains harbouring transposon insertions in two such genes, toxR (a toxin regulator) and mshA (the 11th gene of the 16-gene mannose-sensitive hemagglutinin (MSHA) type IV pilus operon), had reduced infectivity of A. cytherea. Deletion of toxR and the MSHA operon in a second strain of V. coralliilyticus, OCN008, that induces acute Montipora white syndrome in a temperature-independent manner had similarly reduced virulence. This work provides a link between temperature-dependent expression of virulence factors in a pathogen and infection of its coral host.

The trpE gene negatively regulates differentiation of heterocysts at the level of induction in Anabaena sp. strain PCC 7120.

Levels of 2-oxoglutarate (2-OG) reflect nitrogen status in many bacteria. In heterocystous cyanobacteria, a spike in the 2-OG level occurs shortly after the removal of combined nitrogen from cultures and is an integral part of the induction of heterocyst differentiation. In this work, deletion of one of the two annotated trpE genes in Anabaena sp. strain PCC 7120 resulted in a spike in the 2-OG level and subsequent differentiation of a wild-type pattern of heterocysts when filaments of the mutant were transferred from growth on ammonia to growth on nitrate. In contrast, 2-OG levels were unaffected in the wild type, which did not differentiate under the same conditions. An inverted-repeat sequence located upstream of trpE bound a central regulator of differentiation, HetR, in vitro and was necessary for HetR-dependent transcription of a reporter fusion and complementation of the mutant phenotype in vivo. Functional complementation of the mutant phenotype with the addition of tryptophan suggested that levels of tryptophan, rather than the demonstrated anthranilate synthase activity of TrpE, mediated the developmental response of the wild type to nitrate. A model is presented for the observed increase in 2-OG in the trpE mutant.

Vibrio coralliilyticus strain OCN008 is an etiological agent of acute Montipora white syndrome.

Identification of a pathogen is a critical first step in the epidemiology and subsequent management of a disease. A limited number of pathogens have been identified for diseases contributing to the global decline of coral populations. Here we describe Vibrio coralliilyticus strain OCN008, which induces acute Montipora white syndrome (aMWS), a tissue loss disease responsible for substantial mortality of the coral Montipora capitata in Kane'ohe Bay, Hawai'i. OCN008 was grown in pure culture, recreated signs of disease in experimentally infected corals, and could be recovered after infection. In addition, strains similar to OCN008 were isolated from diseased coral from the field but not from healthy M. capitata. OCN008 repeatedly induced the loss of healthy M. capitata tissue from fragments under laboratory conditions with a minimum infectious dose of between 10(7) and 10(8) CFU/ml of water. In contrast, Porites compressa was not infected by OCN008, indicating the host specificity of the pathogen. A decrease in water temperature from 27 to 23°C affected the time to disease onset, but the risk of infection was not significantly reduced. Temperature-dependent bleaching, which has been observed with the V. coralliilyticus type strain BAA-450, was not observed during infection with OCN008. A comparison of the OCN008 genome to the genomes of pathogenic V. coralliilyticus strains BAA-450 and P1 revealed similar virulence-associated genes and quorum-sensing systems. Despite this genetic similarity, infections of M. capitata by OCN008 do not follow the paradigm for V. coralliilyticus infections established by the type strain.