Berkshire, Buckinghamshire & Oxfordshire Wildlife Trust (BBOWT) has this year recorded the highest number of glow worms ever at a nature reserve.
The Trust also saw record amounts of orchids at their nature reserves, as well as a rare golden hoverfly and goshawks breeding at a woodland for the first time.
Colin Williams, senior ecology officer at the Trust, said: "I am so pleased with some of the fantastic wildlife we've recorded on our nature reserves this year.
"Getting record numbers of glow worms or seeing new dragonflies is great - but this is almost entirely down to decades of incredible work by our staff and thousands of tireless unpaid volunteers."
Mr Williams added: "However the wider picture for wildlife in our three counties is incredibly distressing, and nature is in crisis across our region.
"I've been at BBOWT for 15 years, and this has been another year that shows the sad decline of wildlife."
303 glow worms were recorded at the BBOWT's Whitecross Green Wood nature reserve, revealing the success of staff and volunteer efforts in nurture the perfect grassland habitat for these creatures.
The same nature reserve also noted a rare sighting of a southern migrant hawker dragonfly laying eggs, marking the first recorded breeding of this species in Oxfordshire.
In the Dancersend reserve in Buckinghamshire, a golden long-horned hoverfly was spotted by volunteer invertebrate expert Sue Taylor.
This marks the first sighting of this specie at the site, and one of the few in the county.
The Trust's Homefield Wood reserve near Marlow had a notable "excellent year" following record-breaking orchid sightings, with as many as 1,111 rare military orchids being seen.
However, the survey results were not all positive.
Chief executive, Estelle Bailey said: "Wildlife across our three counties is in crisis and desperately needs our help."
The Trust recorded a reduction in the hazel dormice numbers, previously recorded at over 100 in 2004, to just two this year.
Butterflies also had an average or below-average year in BBOWT's surveys.
Mr Williams said: "The decline in butterflies is caused by a host of human actions such as destruction of their habitat and pollution.
"The warmer winters we are now getting mean the frosts that are essential to kill parasites that predate butterfly eggs are not happening enough."
The Foxholes Wood nature reserve near Chipping Norton observed a pair of breeding goshawks for the first time.
BBOWT recently started its most ambitious fundraising appeal, the Nature Recovery Fund.
The appeal aims to raise £3 million over the next three years to fund every aspect of the Trust's work on nature and climate crisis.
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