Acta Limnologica Brasiliensia
http://www.alb.periodikos.com.br/article/doi/10.1590/S2179-975X7624
Acta Limnologica Brasiliensia
Seção Temática: Métodos

Effects of freezing on the carbon and nitrogen stable isotopic compositions of fish tissues

Efeitos do congelamento nas composições isotópicas estáveis de carbono e nitrogênio nos tecidos de peixes

Luciana Mulero Beloto; Thiago Simon Marques; Barbara Protocevich; Ronnie Von Mateus Ferreira; Plinio Barbosa de Camargo

http://dx.doi.org/10.1590/S2179-975X7624 Acta Limnol. Bras. (Online), vol.37, e32, 2025
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Abstract

Stable isotope analysis is a vital tool for elucidating nutrient flow within food chains. Studies utilizing this methodology must consider potential variables that could influence the δ13C and δ15N values of the species being examined. One such variable is the preservation of samples in a frozen state during field studies, particularly when immediate sample processing is not possible.

Aim: The objective of this study is to investigate the long-term effects of freeze-thaw at intervals of 1, 2, 7, 15, 30, 90 and 180 days on the δ13C, δ15N, and C:N ratios in biological samples from three fish species.

Methods: Tissues from three estuarine fish species (Cathorops spixii, Genidens barbus and Chloroscombrus chrysurus) were frozen and aliquots were subjected to thawing for sample collection aimed at isotopic determination.

Results: The discrepancies observed in this study were related to the δ13C and C:N ratios in the skin samples of C. spixii and G. barbus, as well as the muscle samples of C. chrysurus. These variations seem to be more closely linked to the potential volatility of the 12C isotope within the samples, resulting in δ13C enrichment and an increase in C:N values.

Conclusions: This suggests that these changes are not solely attributable to the freeze-thaw process, but may also be influenced by other factors like the lipid oxidation. Our findings suggest that this process significantly influences the δ13C and C:N ratios of skin and muscle tissues. Furthermore, we advocate for prompt processing of samples to minimize the impacts of freeze-thaw events.

Keywords

marine fish; experiment; preservation of sample; ratios; isotopes

Resumo

A análise de isótopos estáveis é uma ferramenta essencial para elucidar o fluxo de nutrientes nas cadeias alimentares. Estudos que utilizam essa metodologia devem considerar as variáveis potenciais que podem influenciar os valores de δ13C e δ15N das espécies analisadas. Uma dessas variáveis é a preservação das amostras em estado congelado durante os estudos de campo, especialmente quando o processamento imediato das amostras não é possível.


Objetivo: O objetivo deste estudo é investigar os efeitos a longo prazo o congelamento e descongelamento em intervalos de 1, 2, 7, 15, 30, 90 e 180 dias sobre os valores de δ13C, δ15N e as razões C:N em amostras biológicas de três espécies de peixes.

Métodos: Tecidos de três espécies de peixes estuarinos (Cathorops spixii, Genidens barbus e Chloroscombrus chrysurus) foram congelados e alíquotas foram submetidas ao descongelamento para coleta de amostras destinadas à determinação isotópica.

Resultados: As discrepâncias observadas neste estudo estavam relacionadas aos valores de δ13C e às razões C:N nas amostras de pele de C. spixii e G. barbus, bem como nas amostras de músculo de C. chrysurus. Essas variações parecem estar mais fortemente ligadas à potencial volatilidade do isótopo 12C nas amostras, resultando em um enriquecimento de δ13C e um aumento nos valores de C:N.

Conclusões: Isso sugere que essas alterações não são exclusivamente atribuíveis ao processo de congelamento e descongelamento, mas também podem ser influenciadas por outros fatores presentes nas amostras. Nossos resultados indicam que esse processo influencia significativamente os valores de δ13C e as razões C:N nos tecidos de pele e músculo. Além disso, recomendamos o processamento imediato das amostras para minimizar os efeitos do congelamento e descongelamento.

Palavras-chave

peixes marinhos; experimento; preservação de amostras; razões; isótopos

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Submetido em:
29/08/2025

Aceito em:
14/10/2025

Publicado em:
17/12/2025

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