Cyclists at the Giro d’Italia have been warned about urinating in bottles during the three-week stage race.

The 109th edition of the Italian Grand Tour began on May 8 and finishes on May 31 after 3,459 kilometres across 21 days of racing.

Monday marked the first of two rest days — but Sunday evening brought a statement from the UCI, cycling’s world governing body, and the race organisers about riders not urinating in their bottles and discarding them.

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It said: “To respect the image of cycling and the Giro d’Italia, the Organiser and the Commissaires’ Panel inform all riders that urinating into a bottle and subsequently discarding it is strictly prohibited.”

It has not been confirmed which riders were responsible for the warning being issued.

But following Sunday’s ninth stage, penalties were handed out for urination and “unseemly behaviour”.

The punishments dished out consisted of fines to the respective teams.

Persistent rule breaking could lead to more serious penalties such as time deductions, point deductions, or even disqualification from the race.

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Portuguese rider Afonso Eulalio currently leads the overall classification by two minutes and 24 seconds.

However, he is not expected to keep the leader’s pink jersey until the final stage in Rome.

Two-time Tour de France winner Jonas Vingegaard is the heavy favourite to claim his first Giro title having won stages seven and nine.

If successful, Dane Vingegaard would become just the eighth male rider to win all three Grand Tours — the Giro d’Italia, Tour de France and Vuelta a Espana.

The others are Jacques Anquetil, Eddy Merckx, Felice Gimondi, Bernard Hinault, Alberto Contador, Vincenzo Nibali and Chris Froome.