Queen conch (Lobatus gigas) is a marine gastropod endemic to the Caribbean. This species is a cultural symbol, being a significant local food source and the second largest commercial fishery in the region. However, over-exploitation and natural habitat degradation have exerted high survival pressure on this species. This work aims to provide novel proteomic data to highlight the metabolism of the species and to provide an important tool for the understanding of queen conch biology and physiology. Herein, we profiled the whole proteome from 3 organs (gills, digestive gland and muscle) of L. gigas combining gel-free and gel-based techniques. Overall 420 clusters of proteins were identified corresponding to the minimum identification requirement of protein sequence redundancy. Gene ontology and KEGG analysis highlighted 59 metabolic pathways between identified proteins. The most relevant routes according to the number of sequences found per pathway were purine and thiamine metabolism, closely related to nucleotide and carbohydrate metabolism. We also emphasize the high number of proteins associated to the biosynthesis of antibiotics (93 proteins and a total of 28 enzymes), which were among the top-twenty pathways identified by KEGG analysis. The proteomics approach allowed the identification and description of putative markers of oxidative stress, xenobiotic metabolism, heat shock response and respiratory chain for the first time in the species, which could be extremely useful in future investigations for diagnosing and monitoring L. gigas population health.

Early diagnosis of endometritis in dairy cattle is currently requires invasive techniques and specialist expertise. The goal of this study is to utilize a gel-free mass-spectrometry based proteomics approach to compare the plasma proteome of dairy cattle with cytological endometritis to those without. Blood samples were collected from cows (N = 112) seven days postpartum (DPP). Plasma samples from a cohort of 20 animals with cytological endometritis (n = 10) and without (n = 10) as classified 21 DPP were selected for proteomic analysis. Differential abundances of proteins between the two animal groups were determined using both fold change (>= 1.5 fold change) and statistical significance threshold (p < .05). A total of 181 non-redundant proteins were quantified, and 25 proteins were found with differential abundance. These include 4 binding protein alpha and mannose binding lectin 2 involved in immune responses. Differentially abundant proteins between the animals were then processed using PANTHER for gene ontology. Gene ontology included associations with innate immune processes, acute phase responses and immune regulation. A potential marker for disease identified here is the "uncharacterized protein G5E513," a protein previously defined by RNA-transcripts. These proteins may form the basis for endometritis prognosis, the development of which is proceeded by systemic changes in immune function. Significance: Endometritis is a costly reproductive disease of lactating dairy cows that warrants timely diagnosis. We utilized a gel-free mass-spectrometry based proteomics approach to compare the plasma proteome of dairy cattle with cytological endometritis to those without, for the characterization of changes in the proteomic profile associated with uterine disease postpartum. Furthermore, we compared the plasma proteome of healthy and affected cows in the same physiological status of production to better understand the relationship between changes in expression of circulating proteins and to unravel essential biological mechanisms involved in bovine cytological endometritis.

Seasonal weight loss (SWL) is a primary constraint for farmers in the Mediterranean and tropics. One cost-effective solution to SWL is utilizing breeds like the Damara sheep that have adapted to deal with nutritional stress. Previous studies concluded that one of the adaptation mechanisms of SWL is a specialized fatty acid metabolism. Accordingly, hepatic-mitochondrial proteomes were compared across two different breeds (24 sheep total, Merino, n = 12 and Damara, n = 12) and two different diets (restricted vs unrestricted diet, 6 per breed, per diet, 24 total). Mitochondrial-proteins were isolated and relatively quantified using Blue native PAGE / 2D-electrophoresis and then analyzed via mass spectrometry. The tool ReviGO summarized the proteomes' gene-ontology terms. A total of 50 proteins were identified with 7 changing significantly in abundance (ANOVA p-value<0.05). Specific abundance patterns of corticosteroid and inflammatory response-associated proteins such as annexin and glutamate dehydrogenase suggests that the Damara has an unusual inflammation response when subjected to SWL in addition to its unique metabolism. All significant proteins warrant further study; Annexin in particular shows promise as a potentially useful biomarker.

The primary objective of this study was to conduct an initial analysis of the proteome of exhaled breath condensate or blow in aquarium-based bottlenose dolphins (Tursiops truncatus) and, secondarily, to determine the commonality of proteins identified in blow with those in plasma of the same animals as recently documented. Exhaled breath condensate was collected from four young (2 to 6 y old), male dolphins using a 50-mL Falcon tube held above the blowhole for ten sequential exhalations; total volume ranged from 60 to 122 mu L. Subsequent to analysis of total protein, 20 mu g of protein from each dolphin was separated by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SUS-PAGE). Of the four samples, three produced sufficient resolution of 15 bands that were excised from respective gels and subject to liquid chromatography-tandem mass spectrometry (LC-MS/MS) and used for protein identification. Mass spectra data were used to search the National Center for Biotechnology Information (NCBI) database restricted to all mammalian proteins. Based on proteins having >= 2 peptides and present in at least two of the three dolphins, a total of 220 blow proteins were identified. While a majority (38 to 51%) of proteins could not be categorized, gene ontology indicated protein binding (26%), cytoplasm constituents (16%), and immune response (16%) dominated the molecular function, cellular component location, and biological process domains, respectively. From noncontemporaneous samples, NCBI Accession numbers of 220 blow proteins described herein and 196 plasma proteins previously identified by LC-MS/MS in the same dolphins were matched. Results indicated a commonality of 21 proteins (5%), with ten (48%) related to the immune system (e.g., complement- and immunoglobulin-related proteins) and the remainder to other various biological systems. Although preliminary, the novelty of these results provides additional support that exhaled breath condensate can be a relevant, less invasive alternative to blood collection to assess or monitor physiological health and pathological states. More specifically, the commonality of several immune-related proteins between the circulatory and respiratory systems provides a foundation for future investigations to determine the potential of these blow proteins as biomarkers that may be diagnostic or prognostic of respiratory health in bottlenose dolphins and, perhaps, other cetaceans.

The effect of feed restriction on the liver protein profiles of two different breeds of sheep was studied. We compared Merino with the Damara, breeds with respectively low and high tolerance to nutritional stress. Each breed was grouped into two nutritional treatments: restricted (12-14% loss of live weight) and control (maintenance). The trial lasted 42 days. Animals were sacrificed and liver samples subjected to label free shotgun proteomics. The resultant proteins had both their fold change and statistical significance in an unpaired t-test calculated to identify differential protein abundance. The tool WebGestalt was utilized to perform an Overrepresentation Enrichment Analysis (ORA) for gene ontology terms associated with the significant proteins. We further validated shotgun proteomics findings using a selected reaction monitoring (SRM)-based targeted proteomics approach, where similar trends in regulation were obtained for a subset of relevant proteins across an independent cohort of animals. Results confirm that Damara has adapted to nutritional stress by mobilizing stored fatty acids within adipose tissue and converting them to energy more efficiently than Merino. Finally, Merino had an overabundance pattern primarily directed to protein synthesis pathways. Regulated proteins identified may be used as a basis for marker selection towards tolerance to nutritional stress. Biological significance: Sheep are one of the most important livestock animals. They remain however poorly understudied and described. Seasonal weight loss (SWL) due to pasture scarcity during the dry season is one of the most limiting conditions to ruminant production in the tropics. To counter SWL, farmers may use supple. mentation, expensive or difficult to implement. A more suitable long-term solution would be to use breeds that are naturally adapted to nutritional stress. This work contrasts two breeds with different levels of tolerance to SWL, the Damara and the Merino, respectively well and poorly adapted. Comparison is conducted at the level of the hepatic tissue and using label free proteomics. This work identifies a series of pathways in the liver of the Damara via label free proteomics that suggest a unique fatty acid metabolic process within this breed. Proteins that have increased abundance in the Damara in association with fatty acid metabolism may be used as potential markers of tolerance to nutritional stress. This research will pave the way for more viable, long-term solutions for farmers facing annual production problems due to drought in the tropics and the Mediterranean region.

This study was designed to initially characterize and evaluate the proteome and other biochemical factors in plasma samples collected in association with performance-based physical activity involving four young male aquarium-based bottlenose dolphins (Tursiops truncates). Blood samples were collected from the tail flukes approximately 15 min before (N = 4 samples) and 15 min after (N = 4 samples) the first public swim interaction of the day, which was reinforced with a daily scheduled feeding regimen and individually prescribed diets. Plasma extracted and trypsin-treated protein extracts were subjected to liquid chromatography-tandem mass spectrometry (LC-MS/MS) and conventional clinical biochemistry analysis. Mass spectra data were, used to search the National Center for Biotechnology Information (NCBI) database restricted to. Tursiops truncates. The search resulted in the identification of 196 unique proteins with a broad range of functional roles based on gene ontology (GO) analysis. Differential regulation of proteins was based on log mean fold change (FC) and statistical probability such that the abundance of lysozyme (FC -1.2036; p < 0.058), an immune-related protein, and Flavin reductase (FC-0.9702; p < 0.004), a metabolic-related protein, were highest before compared to after the swim interaction; both proteins decreased 58 and 52%, respectively. Correspondingly, 15 known proteins and other biochemical factors were highest before compared to after the swim interaction. Glucose, creatinine, alkaline phosphatase, blood urea nitrogen, calcium, and magnesium decreased 3 to 26% (p < 0.004 to 0.07). In conclusion, down regulation of immune- and metabolic-related proteins and multiple other biochemical factors after the swim interaction may represent a homeostatic response to high values in the absence of substantial food in-between the daily feeding regimen or in anticipation of food before the first swim interaction of the day. Thereafter, low values may represent a response to food consumption or satisfaction. The novelty of these initial results suggests that performance-based physical activity is not immunologically or metabolically challenging in bottle-nose dolphins conditioned for swim interactive programs; however, future studies are required for clarification.