Investigating an increase in Florida manatee mortalities using a proteomic approach.

Two large-scale Florida manatee (Trichechus manatus latirostris) mortality episodes were reported on separate coasts of Florida in 2013. The east coast mortality episode was associated with an unknown etiology in the Indian River Lagoon (IRL). The west coast mortality episode was attributed to a persistent Karenia brevis algal bloom or 'red tide' centered in Southwest Florida. Manatees from the IRL also had signs of cold stress. To investigate these two mortality episodes, two proteomic experiments were performed, using two-dimensional difference in gel electrophoresis (2D-DIGE) and isobaric tags for relative and absolute quantification (iTRAQ) LC-MS/MS. Manatees from the IRL displayed increased levels of several proteins in their serum samples compared to controls, including kininogen-1 isoform 1, alpha-1-microglobulin/bikunen precursor, histidine-rich glycoprotein, properdin, and complement C4-A isoform 1. In the red tide group, the following proteins were increased: ceruloplasmin, pyruvate kinase isozymes M1/M2 isoform 3, angiotensinogen, complement C4-A isoform 1, and complement C3. These proteins are associated with acute-phase response, amyloid formation and accumulation, copper and iron homeostasis, the complement cascade pathway, and other important cellular functions. The increased level of complement C4 protein observed in the red tide group was confirmed through the use of Western Blot.

Investigating the gene expression profiles of rehabilitated Florida manatees (Trichechus manatus latirostris) following red tide exposure.

To investigate a Florida manatee (Trichechus manatus latirostris) mortality event following a red tide bloom in Southwest Florida, an RNA sequencing experiment was conducted. Gene expression changes in white blood cells were assessed in manatees rescued from a red tide affected area (n = 4) and a control group (n = 7) using RNA sequencing. The genes with the largest fold changes were compared between the two groups to identify molecular pathways related to cellular and disease processes. In total, 591 genes (false discovery rate <0.05) were differentially expressed in the red tide group. Of these, 158 were upregulated and 433 were downregulated. This suggests major changes in white blood cell composition following an exposure to red tide. The most highly upregulated gene, Osteoclast associated 2C immunoglobulin-like receptor (OSCAR), was upregulated 12-fold. This gene is involved in initiating the immune response and maintaining a role in adaptive and innate immunity. The most highly downregulated gene, Piccolo presynaptic cytomatrix protein (PCLO), was downregulated by a factor of 977-fold. This gene is associated with cognitive functioning and neurotransmitter release. Downregulation of this gene in other studies was associated with neuronal loss and neuron synapse dysfunction. Among the cellular pathways that were most affected, immune response, including inflammation, wounds and injuries, cell proliferation, and apoptosis were the most predominant. The pathway with the most differentially expressed genes was the immune response pathway with 98 genes involved, many of them downregulated. Assessing the changes in gene expression associated with red tide exposure enhances our understanding of manatee immune response to the red tide toxins and will aid in the development of red tide biomarkers.

Using the Electronic Foodborne Outbreak Reporting System (eFORS) to improve foodborne outbreak surveillance, investigations, and program evaluation.

Challenges exist in comparing foodborne disease outbreaks (FBDOs) across states due to important differences in reporting practices and investigations. Variables such as FBDO size, population size, number of tourists, and suspected etiology are important to consider when interpreting FBDO data. Analysis of eFORS data can be valuable in improving state FBDO investigations. From 2000 to 2005, Florida reported a greater proportion of FBDOs, with two cases per outbreak, than the U.S. as a whole (40.4% in Florida vs. 17.2% in the U.S.). Reporting a higher rate of small FBDOs provided more opportunities for public health interventions but contributed to a lower agent confirmation rate (17.0% in Florida vs. 42.2% in the U.S.). While the Electronic Foodborne Outbreak Reporting System's (eFORS) database brought great improvements in national FBDO surveillance, as with any complex surveillance system, considerable knowledge and specialized expertise is required to properly analyze and interpret the data, especially because there is a large variation in state reports to eFORS.

Cryptosporidiosis outbreak in a Nassau County, Florida, return travel group from Ireland, May 24, 2006-June 4, 2006.

The Nassau County Health Department (NCHD) in Florida investigated an outbreak of gastrointestinal (GI) illness in a returning choral group who toured Ireland from May 24 to June 4, 2006. The travel group, consisting predominantly of retirees, had performed at several churches and at a dinner theater in Ireland. The NCHD administered a telephone questionnaire to 40 of the 41 group members to examine possible water exposures; common meals; and food, travel, and clinical histories. The results of the questionnaire showed that 29 people met the case definition for the outbreak. Five stool samples from travel group members tested positive for Cryptosporidium parvum, a species that is animal in origin and often spread through an environmental contamination with animal feces. All five positive samples were subtyped 11aA16G1R1b, a strain that scientists at the Centers for Disease Control and Prevention (CDC) Division of Parasitic Diseases detected twice in 2006 in other human specimens from Northern Ireland.