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FISH ODOUR TRIMETHYLAMINE MIGHT BE A GOOD

CANDIDATE FOR TESTING GENDER DEPENDENCE IN
HUMAN OLFACTION

L’ODORE DI PESCE TRIMETILAMMINA POTREBBE

ESSERE UN BUON CANDIDATO PER DIMOSTRARE LA
DIPENDENZA DAL SESSO DELLA CAPACITA’ OLFATTIVA
NELL’UOMO

BIGNETTI1 Enrico, AIELLO2 Gaetano L., SINESIO3 Fiorella and CANNELLA4 Carlo

Keywords.
Olfaction, gender, trimethylamine, trimethylaminuria, fish consumption.

Parole chiave.

Olfatto, sesso, trimetilammina, trimetilamminuria, consumi di pesce.

Abstract.

One major question was whether odors are perceived by women and men in
the same way or not. A strong evidence in the literature described a wide variation
of olfactory sensitivity in women during menses. Several explanations were given
to account for the variation, none of them convincing. Within the cycle, the greatest
attention was posed only on the olfactory sensitivity increase during the follicular
phase as an efficient odor-based mating strategy. Up to us, the attention should be
posed also towards the mechanism of worsening of the sensitivity during bleeding
days. In this cycle phase, malodorous discharges through body fluids, e.g. trimethyl-
aminuria, are maximal. Thus, olfactory threshold regulation across the cycle might
represent either a physiological mechanism to search sexual partner and a defensive
mechanism to avoid a possible self-refusal. Testosterone and estradiol might be pri-
marily responsible for this complex regulation. The fluctuation of olfactory thresh-
olds might influence also alimentary habits especially towards Fish consumption.

1 Prof. of Clinical Biochemistry and Molecular Biology at the Faculty of Vet. Med. of Parma. Ad-
dress: Lab. “Food Neurochemistry”, Dept. of Physics, University of Parma, Via G.P. Usberti 7a, 43100
Parma, Italy (email: enrico.bignetti@unipr.it)
This work was done in conjunction with INRAN to start up a preliminary study on fish consumption in
Italy.
2 Dept. of Physics & Related Technologies (DIFTER), University of Palermo, Viale delle Scienze, Ed.
#18, 90128 Palermo, Italy
3 “National Research Institute for Food and Nutrition” (INRAN), Viale Ardeatina 546, 00178 Roma,
Italy.
4 Prof. at Sapienza University and President of “National Research Institute for Food and Nutrition”
(INRAN), Viale Ardeatina 546, 00178 Roma, Italy.

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Our hypothesis might be tested by carrying out psychophysical tests that use the fish
odour trimethylamine as odor model.

Riassunto

Un lavoro recente sembra indicare che la capacità olfattiva delle donne du-
rante il ciclo vari sorprendentemente di alcuni ordini di grandezza, dalla fase folli-
colare (massima) alla mestruale (minima). Durante la fase mestruale, pare anche che
le donne vadano soggette alla perdita di trimetilamina (l’odore sgradevole di pesce),
attraverso i liquidi corporei ed in particolare attraverso le urine. Dunque, questo la-
voro propone l’ipotesi che la fluttuazione dell’olfatto nel ciclo sia un meccanismo
fisiologico per una migliore strategia di ricerca sessuale, da un lato, e di auto-difesa
dal disgusto su base olfattiva del proprio corpo, dall’altro. Testosterone ed estradiolo
potrebbero essere coinvolti in questo complesso meccanismo fisiologico. Un test che
utilizzi la Trimetilammina come odore modello, potrebbe dare una risposta definitiva
all’ipotesi di meccanismo proposta. Con questo si potrebbero aprire nuove linee di
ricerca sia sulle strategie di accoppiamento nell’uomo e sul confronto delle attitudini
alimentari tra uomo e donna.

Introduction

Volatile chemical signals play a crucial role in human behaviour associated

with social interaction, comunication and feeding. As a matter of fact, humans can
discriminate a thousand of odorants within an extraordinary vast landscape of syn-
thetic and natural odorants [1].
One major question was whether odors are perceived by women and men
in the same way or not [2-6]. The question as it might provide an answer, once for
all, to the question whether women and men do have equal chances to achieve social
and professional goals. Knowing whether men and women exhibit the same aver-
sion against, or preference for seafood based on their olfactory discriminants might
provide precious hints about the alimentary habits of a family, and/or push a subject
towards a professional carreer as a chef.
The sensory modalities by which fish is perceived by people, beside produ-
cing a specific attitude towards a diet, might reveal several psychological constructs,
such as beliefs, interest, motivation, even social norms, knowledge and other beha-
viours. All this might have a relevance to social sciences and food marketing.
As an example, Olsen [7] shows there is a positive relationship between age
and frequency of seafood consumption. This, and other examples, pointed our atten-
tion to the need to explore how fish smell is perceived by humans of both sexes, at
any stage of their life. Below, we will critically review the state of the art of gender-
dependence in human olfaction, on one side, and some TMA features, on the other.
Then, a series of tests will be proposed which might reveal hidden aspects of this
particular sensory modality.

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Gender-dependence in olfaction
The question whether human olfactory threshold could depend on sex and
age was addressed by several authors. Two volatile C-19-steroids, which are conside-
red putative pheromones in humans, were found [8-9] to contribute to mood, memory
and autonomic system responses in a gender-specific manner. A sex-specific differen-
ce in olfactory sensitivity for these two molecules was demonstrated also in primates.
A clear-cut example of sex-specific bimodal olfactory sensitivity was demonstrated
in spider monkeys, with male responding to one molecule only and female to the
other [6]. However, these molecules carry a well-defined physiological information
targeted mainly to sex behaviour, with scarce relevance to the mechanism of odorant
perception and recognition.
By studying specific anosmia to “primary odorants”, Amoore came to the
conclusion that there was a logarithmic deterioration in olfactory perception with
age, which was unaffected by sex [2]. The results of the experiments on olfaction,
however, are always open to debate. One reason is the intrinsic complexity of the
olfactory system, and the lack of standardization of the methods. An emblematic case
is reported by Öberg et al. [10]. By using a large sample of women and men, these
authors investigated gender-related differences in several tasks aimed to study: abso-
lute sensitivity, intensity discrimination, quality discrimination, episodic recognition
memory, and identification. Although a better performance was registered by female
subjects on episodic odor memory capability towards familiar odors, no sex related
differences in sensory acuity were observed.
Collectively, by looking at these experiments, one can always find signifi-
cant drawbacks in the methodologies and protocols used, like large hormonal disho-
mogeneity within subjects in the same sample, which might cause large intraspecific
error. If the sample is made of women, one should consider also the menstrual cycle
as a crucial physiological parameter. Circadian hormonal oscillations (corresponding
to: menstrual, follicular, ovulatory and luteal or progestagenic phases) are known to
profoundly alter her normal lifestyle. In other words, a different way of perceiving
odors during the cycle may affect some abitual choices.
Up to us, the examples reported in the literature untill now exhibited a large
uncertainty in data collection and interpretation. This might stand on two main rea-
sons: either the data collection was rather scarce from a statistical point of view or a
bias prompted by an erroneous model prevision was causing a vitious data interpre-
tation.
In a recent paper, some authors [5] faced olfaction gender-dependence at le-
ast in a statistically-convincing manner. They monitored olfactory thresholds towards
the banana-like odor (amyl-acetate) in different groups of subjects. It was evident that
the threshold varied significantly across the menstrual cycle. In particular, women
exhibited the lowest olfactory thresholds during the ovulatory phase, and the highest
during the bleeding days.
Several explanations were tentatively given by the authors to account for
a variation of the threshold with the phase of the menstrual cycle. None of them,
however, was convincing. As a conclusion, the effect of the cycle on the olfactory
threshold is either a by-product of other parallel physiological events, or it hides an

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important biological event whose teleological meaning is still unclear.

This second hypothesis seems us much more appealing. Thus, we argue the
finalties why olfactory sensitivity for the banana-like odor is low during the men-
strual phase, with median threshold running about 30 times higher than in the ovula-
tory phase, and about 10-time lower than in control men. The reason might lie deep
inside those very mechanisms that specifically differentiate women from men.

Trimethylamine: an odor that might unveil the question of olfactory sensitivity

changes in cycling women
Methylamine, Dimethylamine and Trimethylamine are derivatives of am-
monia with a typical rotten fish odor at lower concentration and of ammonia-like
odor at higher concentrations. Human studies conducted to assess odor perception
of these compounds demonstrated that all three are considered maleodorant, and can
be perceived dissolved in water at rather high dilutions, TMA being the one with a
threshold 3-4 orders of magnitude lower than the others [11]. A significant amount of
human subjects exhibited a pronounced anosmia to the low-molecular-weight tertiary
amines (a specific TMA anosmia was monitored in about 7% of them) and in a lesser
degree to primary and secondary amines [12].
The above substances are present in the decomposition of plants and animals
as microbial degradation of animal byproducts. As far as fish is concerned, TMA de-
rives from enzymatic reduction of the non-volatile precursor Trimethylamine Oxide
(TMAO), carried out by bacteria on the skin [13]. Among the cultures isolated from
spoiling fish, Shewanella putrefaciens were identified as the major agent responsible
of TMA and off-odour production [14]. Bacterial spoilage and associated malodor
production in fish can cause a natural avoidance behaviour of the consumers. Thus,
controlling these processes was of primarily importance for industrial fish meal plants.
On one end, new packaging methods and new additives were proposed in mitigation
strategies, succesfull even at long distances from the industrial plants (Chaen et al.,
“http://www.freepatentsonline.com/6576281.html”). On the other end, the research
tried to further improve the ways of detecting fish odors instrumentally, with detec-
tion thresholds down to human nose. To this aim, a TMA gas sensor was developed
and utilized to detect fish freshness [15]. More recently, specific chemical analyses of
malodorous compounds downwind of a fish meal were carried out by means of gas
chromatography in tandem with mass spectrometry, and the results compared with
human panelist data [16].
TMA can be found not only on spoiling fish but also in gastric juices, in the
saliva and in blood of humans and other mammalians. It can be considered as a nor-
mal constituent of mammalian wastes like feces, urines and exhaled air. The amount
normally excreted can be partially influenced by the diet, especially when it contains
fish [17].
More rarely, the excretion may become abnormal depending on specific pa-
tological conditions. Striking examples are the cases of “Bacterial Vaginosis” (BV)
(see for a review: [18]) and of “Primary Trimethylaminuria” (also known as “Fish-
odor-syndrome”) [19].
Actually, menstruations in women can be an additional factor causing tran-

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sient trimethylaminuria in self-reported subjects suffering from malodor, and even

in healthy women harboring functionally active monooxygenase. Infact, beside the
autosomal recessive defect producing a totally inactive FMO3, there is also a poli-
morphic landscape of mild mutations of FMO3 gene causing variable degrees of tri-
methylaminuria during menses. The evidence of trimethylaminuria during bleeding
days in women was discovered since long ago [20], but the enzymatic mechanism
that regulate odor discharge was explained only recently by Shimitzu et al. [21].

The conclusive model and the experimental proposal

In conclusion, more or less all cycling women can undergo a peak of malo-
dorous trimethylaminuria coinciding with bleeding days, due to a specific FMO3 de-
ficiency in those days. By observing the dynamics of olfaction and FMO3 in cycling
women, it looked as if these systems were undergoing a common regulatory fate, i.e.
as though both systems were under control of the same hormones regulating menses.
Thus, lowering the olfaction during bleeding days might reflect a self-defensive me-
chanism. Conversely, the extraordinary olfaction switched on at the ovulatory phase,
might be used as a contrivance to refine one of the most sophisticated tool for mating
search. By this, we do not exclusively or necessarily refer to a “simple” interplay
of pheromones but also to other refined strategies based on smell, e.g. the use of an
agreeable perfume, the choice of the partner with the best smell or the avoidance of
other competing women in the proximity etc..
Moreover, the positive FMO3 induction observed in pregnant women might
be easily explained as an adapting reponse of mother’s liver against the need of a
surplus of detoxifying work required by the son.
In conclusion, there might be a hidden functional cross-correlation between
cycling phases, liver detoxification and olfactory threshold variation. These physio-
logical processes, although apparently far from each other since pertaining to very
different anatomical districts, might actually work under the control of a same hor-
monal system.
Now, the question relevant to us is whether we can support these assertions
with experimental evidences. On the basis of the above evidences, it should be fe-
asible to use TMA odor as a probe to reveal whether our model is correct. Up to
now, to challenge the effect of menses on female olfaction it was used amyl-acetate,
a strongly attractive odor. In our opinion, if these results were confirmed also with
TMA, one of the most aversive odor excreted with body fluids during menstruations,
then a further step could be done towards the understanding of cyclic behaviour of
olfaction in women.
The protocol used by Navarrete-Palacios et al. [5] looks the most appropria-
te, mainly because of the large groups engaged in the tests, the odor used, the sniff-
bottle technique, and the procedure of ascending concentrations. Thus, we could
adopt the above protocol almost entirely, except for the replacement of the pleasant
amyl-acetate with the unpleasant TMA.
As mentioned above the mean values of amylacetate olfactory thresholds
in cycling women fluctuate by several orders of magnitude accross the steady value
which was instead observed in men. Therefore, the experiments with TMA as a mo-

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del odour instead of amylacetate, should again be carried out with men as reference
subjects. These results should possibly lead to extrapolate interesting correlations
with fish consumption and in particular with the frequency of it, either in women
and in men. We are aware that such a project will necessary need a multidisciplinary
approach

Acknowledgements

We would like to thank Prof P. Pelosi of the University of Pisa, Italy, for helpful
discussion.

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