Characterization and finding the origin of off-flavor compounds in Nile tilapia cultured in net cages in hydroelectric reservoirs, São Paulo State, Brazil.

An increasing demand for fish products has led to an intensive aquaculture production in Brazil, and cultivation of fish constituted 860 × 103 tons in 2022, contributing to the 87% of total fish consumption. Nile tilapia constitutes almost half of the aquaculture production, and most tilapia farms use floating net cages. One of the major constraints of intensive fish production is production of off-flavors. Release of nutrients by the fish leads to deterioration of the water quality and stimulates growth of microorganisms, also including off-flavor producing species. The objective of this study was to determine levels of taste and odor compounds (geosmin, 2-MIB and a selection of volatile compounds) and their impact on the flavor quality of Nile tilapia produced in net cages in reservoirs in São Paulo State, Brazil. GC-MS analysis of fish and water from six different farms showed concentrations of geosmin in the water from 1 to 8 ng/L, while geosmin in fish flesh ranged from 40 to 750 ng/kg. The level of 2-MIB in water was 2 to 25 ng/L, and 0 to 800 ng/kg fish. The GC-MS analysis also revealed presence of more than 100 volatile organic compounds in the fish flesh, consisting of aldehydes, alcohols, benzene derivatives, hydrocarbons, ketones and few other compounds. Geosmin and 2-MIB related flavor notes were detected in all fish by a sensory panel, and a high correlation between the chemical and sensory analyses was found. The potential impact of the volatile organic compounds on the fish flavor is discussed. Analysis of the water quality in the reservoirs indicated that levels of geosmin and 2-MIB levels were highly influenced by the nutrient levels in the water.

Monitoring of saxitoxin production in lakes in Denmark by molecular, chromatographic and microscopic approaches.

Diversity of phytoplankton in three eutrophic and two mesotrophic lakes in Denmark was determined by microscopic and HPLC pigment analyses to identify and quantify potential saxitoxin (STX) producing cyanobacteria. Potential dominant STX-producers were identified to the filamentous genera Dolichospermum, Cuspidothrix, Phormidium and Planktolyngbya. Presence of STX production was documented by extraction of five intracellular STXs that included (in declining concentration in the cyanobacteria) dc-neo-STX, neo-STX, dc-STX, STX and GTX. Total concentrations of the five STXs varied from 9 to 142 fg per potential STX producer, corresponding to 87 to 985 ng L-1 in the lakes. For molecular detection of the STX-producers, a quantitative PCR method was developed by design of a new robust primer set with broad coverage to target the sxtA gene that is common to all STX-producing cyanobacteria. After validation, copy numbers of the sxtA gene were determined to vary from about 104 (mesotrophic lakes) to 108 per mL (the most eutrophic lake). A moderate but significant correlation was observed between abundance of the sxtA copies and concentrations of the five intracellular STXs. The qPCR assay was found to be a rapid and robust procedure for quantification of STX producers. Saxitoxin and its analogs appeared not to cause health concerns in the lakes, but commercial fishing for pike perch in the most eutrophic lake should be monitored to test for food web accumulation of STXs.

Microbial Production of the Off-Flavor Geosmin in Tilapia Production in Brazilian Water Reservoirs: Importance of Bacteria in the Intestine and Other Fish-Associated Environments.

AIM: To determine major sources of microbially produced geosmin in the commercially important aquaculture fish species tilapia. METHODS AND RESULTS: Abundance and composition of geosmin-producing bacteria in water and fish biosphere (intestine, digesta, and fins) of Nile tilapia (Oreachromis niloticus) raised in net cages in Brazilian freshwater farms were examined. By combining qPCR of the geosmin synthase geoA gene and 16S rRNA gene amplicon sequencing to identify potential geosmin-producing organisms, we observed that the proportion and composition of geosmin producers appeared to be rather similar in the water, digesta, intestinal mucous, and on skin, making up about 0.1-0.2% of the total bacterial densities. A high proportion of Cyanobacteria and other putative geosmin producers affiliated to the Actinomycetales were identified in the intestinal mucous layer. The main uptake site for geosmin in fish is traditionally assumed to be through the gill surface, but the present results suggest that uptake by the intestinal tract may represent a major source of geosmin uptake in fish. CONCLUSION: The high abundance of geosmin-producing bacteria in the intestinal mucous layer and digesta may indicate that the digestive system in fish is an important, but hitherto overlooked, source of geosmin and likely other off-flavors in fish. SIGNIFICANCE AND IMPACT OF STUDY: Tainting of fish by microbially produced off-flavors spoils fish quality and lowers consumer preferences for aquaculture-produced freshwater fish. Our results highlight the potential for the application of probiotic microorganisms for management of the intestinal microflora to improve the fish quality. HIGHLIGHTS: -Off-flavor producing bacteria are widely abundant in aquaculture.-Off-flavor producers found on skin surface of fish.-Off-flavor producing bacteria accumulate in the digestive system.-Off-flavor producers might release significant amounts of off-flavor during lysis in the gut.-Off-flavor uptake through the digestive system might be quantitatively significant.

Conidia-based fluorescence quantification of Streptomyces.

Determination of cell numbers in filamentous bacteria, such as Streptomyces, is challenging due to the tangled and twisted structure of the filaments and formation of cell clumps in liquid cultures. Here, we developed a conidia-based approach, in which fluorescence of conidia, after staining with the DNA-binding stain SYBR Green 1, was related to SYBR Green 1 fluorescence of DNA in Streptomyces. When cell number in Streptomyces filaments, determined by the conidia assay, was compared to number obtained by a qPCR assay, 34 to 62% of cells in the Streptomyces filaments were recovered. The difference in numbers probably reflects an insufficient extraction of DNA from the Gram-positive bacteria, rather than underestimation of the actual cell number by the conidia-based determination. The conidia-based approach appears to be a fast and reliable procedure for counting cell numbers in Streptomyces filaments but it can also be used for other filamentous bacteria, if proper standard curves can be made.

Potential Contribution of Fish Feed and Phytoplankton to the Content of Volatile Terpenes in Cultured Pangasius (Pangasianodon hypophthalmus) and Tilapia (Oreochromis niloticus).

Geosmin and 2-methylisoborneol are the most recognized off-flavors in freshwater fish, but terpenes may also contribute off-flavor in fish. We identified six monoterpenes, 11 sesquiterpenes, and three terpene-related compounds in pangasius and tilapia from aquaculture farms in Bangladesh. The concentrations of most of the volatiles were below published odor thresholds, except for α-pinene, limonene, ß-caryophyllene, α-humulene, and ß-ionone in tilapia, and limonene and ß-ionone in pangasius. To identify sources of the terpenes, terpene profiles of fish feed and phytoplankton in the ponds were analyzed. In feed and mustard cake (feed ingredient), five monoterpenes and two sesquiterpenes were identified, and five of these compounds were also detected in the fish. In phytoplankton, 11 monoterpenes were found and three also occurred in the fish. The higher number of terpenes common to both fish and feed, than to fish and phytoplankton, suggests that feed was a more abundant source of odor-active terpenes in the fish than phytoplankton.