High Frequency of Histamine-Producing Bacteria in the Enological Environment and Instability of the Histidine Decarboxylase Production Phenotype

Abstract

Lactic acid bacteria contribute to wine transformation during malolactic fermentation. They generally improve the sensorial properties of wine, but some strains produce histamine, a toxic substance that causes health issues. Histamine-producing strains belong to species of the genera Oenococcus, Lactobacillus, and Pediococcus. All carry an hdcA gene coding for a histidine decarboxylase that converts histidine into histamine. For this study, a method based on quantitative PCR and targeting hdcA was developed to enumerate these bacteria in wine. This method was efficient for determining populations of 1 to 10 7 CFU per ml. An analysis of 264 samples collected from 116 wineries of the same region during malolactic fermentation revealed that these bacteria were present in almost all wines and at important levels, exceeding 10 3 CFU per ml in 70% of the samples. Histamine occurred at an often important level in wines containing populations of the above-mentioned bacteria. Fifty-four colonies of histamine producers isolated from four wines were characterized at the genetic level. All were strains of Oenococcus oeni that grouped into eight strain types by randomly amplified polymorphic DNA analysis. Some strains were isolated from wines collected in distant wineries. Moreover, hdcA was detected on a large and possibly unstable plasmid in these strains of O. oeni. Taken together, the results suggest that the risk of histamine production exists in almost all wines and is important when the population of histamine-producing bacteria exceeds 10 3 per ml. Strains of O. oeni producing histamine are frequent in wine during malolactic fermentation, but they may lose this capacity during subcultures in the laboratory.

Scientific Opinion on risk based control of biogenic amine formation in fermented foods

“…For example, a multiplex PCR method for the simultaneous detection of LAB with the potential to produce histamine-, tyramine and putrescine-has been described (Marcobal et al, 2005). Furthermore, a quantitative real-time PCR for histamine-producing LAB (Fernández et al, 2006;Lucas et al, 2008) and for tyramine-producing bacteria has also been successfully applied to different stages of cheese manufacture including the final product (Ladero et al, 2008;Ladero et al, 2010b;Gardini et al, 2008). Molecular methods have also been designed to study histidine-and tyrosine-decarboxylase gene expression under conditions relevant for food fermentation (Rossi et al, 2011), including cheese making and sausage fermentation , respectively.…”

Biogenic amines in fermented foods

“…A multiplex PCR method for the simultaneous detection of histamine-, tyramine-and putrescine-producing LAB has recently been proposed in order to identify BA-producing strains in wine and cider (Marcobal et al, 2005). In addition to end-point PCR analysis, a realtime quantitative PCR has been developed for detecting histamine-producing LAB in cheese and wine (Fernández et al, 2006;Lucas et al, 2008) or successfully used in the different steps of cheese manufacture and wine fermentation (Ladero et al, 2008;Nannelli et al, 2008).…”

Malolactic Fermentation: The ABC’s of MLF

“…MorenoArribas et al (2000) identified O. oeni as the main LAB responsible for histamine formation and lactobacilli for tyramine formation. Lucas et al (2008) identified 54 colonies of histamine producing isolates as O. oeni and despite the fact that histamine producing O. oeni are frequently found in wine, it was also found that LAB may lose this ability due to instability of the phenotype. Histamine producing LAB all carry an hdcA gene coding for a histidine decarboxylase (HDC) that converts histidine to histamine.…”