The Portuguese champions emerged triumphant by a 2-1 scoreline in last week's opening encounter, where a pair of second-half Angel di Maria penalties did the damage.
Match preview
As Liverpool rode off into the Europa League sunset, Toulouse's second-placed finish in the group stage would force the Coupe de France holders to jump through one more hoop if they were to prolong their second-tier European journey, but that hurdle may now be too high to overcome.
Carles Martinez's side were ostensibly on course to travel home with an applaudable draw as Mikkel Desler cancelled out Di Maria's first effort from 12 yards, but with six minutes of added time on the clock, the ex-Real Madrid man found the mark from the spot again after Christian Mawissa felled Marcos Leonardo.
Mawissa being made to take the walk of shame for the challenge added insult to injury for a despondent Violets contingent, but the visitors to Stadium de Toulouse are far from home and hosed; Martinez's men took seven points from nine at home in the group phase, gleaning three of them in their memorable 3-2 beating of Liverpool.
Furthermore, Toulouse regained a measure of morale in Sunday's Ligue 1 encounter with Monaco, leaving the Stade Louis II with a 2-1 success to their name, as Logan Costa - culpable of the handball which led to Di Maria's first goal last Thursday - atoned for that mistake with a 70th-minute winner.
However, Martinez's men are now clean sheet-less in six successive matches and have failed to triumph in a single home game since their triumph over the Reds in November, drawing their following three games before falling to a trio of defeats in their most recent Stadium Municipal fixtures.
In that time, Toulouse have made a habit of scoring once and then failing to score in their next home fixture, and with the hosts' last match on familiar soil ending in a 2-1 loss to Nantes, that sequence bodes well for a Benfica side who need only hold what they have.
A matchday six triumph over Red Bull Salzburg came too late to save the Eagles' Champions League bacon - they had failed to win any of their first five group fixtures versus the Austrian champions, Real Sociedad or Inter Milan in the continent's premier competition - but the visitors have never been eliminated from the Europa League knockout round playoffs when winning the first leg.
Benfica's journey did end at this point versus Shakhtar Donetsk in 2019-20 and Arsenal in 2020-21, but neither of those ties saw the Eagles come up trumps in the opening encounter, and Roger Schmidt's men also have the wind in their sails domestically heading into Thursday's return fixture.
Three days after edging past Toulouse the first time, Schmidt's troops demolished Vizela 6-1 in Primeira Liga action - notching five of their efforts before the half-time whistle sounded - to cling onto first place ahead of Sporting Lisbon thanks to the head-to-head metric.
Each of Benfica's last six matches has seen the Eagles notch at least two goals, and the visitors also have a seven-game unbeaten run on the road to boast since going down in November's Champions League tie to Sociedad, but they are yet to test their mettle on Toulouse's turf.
Toulouse Europa League form:
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L
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D
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Toulouse form (all competitions):
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Benfica Europa League form:
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Benfica form (all competitions):
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D
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Team News
By virtue of his sending off for two bookable offences in the first leg, Toulouse defender Mawissa will miss Thursday's reunion through suspension, although the 18-year-old was only introduced in the 70th minute of last week's clash anyway.
Mawissa's disastrous cameo should therefore have no effect on Martinez's starting lineup, especially with left-back Gabriel Suazo returning from a domestic ban in time for the second leg; Mawissa deputised for the Chile international against Monaco.
Denis Genreau (groin) and Zakaria Aboukhlal (knee) are still missing for the hosts, who should also recall first-leg goalscorer Desler to his right wing-back role in place of Warren Kamanzi.
Sticking with the full-back theme, Benfica remain without on-loan Paris Saint-Germain defender Juan Bernat due to the Spaniard's muscular problem, so Manchester United-owned Alvaro Carreras is primed to bomb down the left-hand side once again.
Rafa Silva was withdrawn at half time in the destruction of Vizela, but that alteration was seemingly a tactical decision from Schmidt, who dropped several big-hitters from the first XI at the weekend with the second leg in mind.
Di Maria, Antonio Silva, Fredrik Aursnes and Arthur Cabral should be among those reinstated to the hosts' starting lineup on Thursday, while midfield fulcrum Orkun Kokcu is available again after a Primeira Liga suspension.
Toulouse possible starting lineup:
Restes; Costa, Nicolaisen, Diarra; Desler, Spierings, Sierro, Suazo; Donnum, Dallinga, Gboho
Benfica possible starting lineup:
Trubin; Aursnes, Silva, Otamendi, Carreras; Neves, Kokcu; Di Maria, Rafa, Neres; Cabral
We say: Toulouse 1-2 Benfica (Benfica win 4-2 on aggregate)
Sitting back and waiting for the right moment to strike should not be the approach Toulouse take on Thursday evening, where the French side must go for broke lest they exit the Europa League with a whimper.