All of the new cases are linked to previously reported cases and 11 were in quarantine throughout their entire infectious period.
Victorian Health Minister Martin Foley urged regional Victorians to stay alert for symptoms in a press conference on Saturday.
"There have been new regional detections in the Shepparton regions," he said.
"With the catchment includes the suburbs of Shepparton, Kialla and small parts of both Shepparton north and Orrvale."
Anyone who lives, works in or has visited Shepparton from August 9 to 11 is urged to watch for the "slightest symptoms" of COVID-19 and get tested if any symptoms arise.
There are also positive wastewater detections in Lakes Entrance, metropolitan Melbourne in the Camberwell and Balwyn area and the western suburbs.
Health authorities said the detections were "significant" neither Shepparton nor Lakes Entrance had any active COVID-19 cases or current exposure sites.
"People in areas with recent wastewater detections need to be especially vigilant in getting tested should they develop even the mildest of symptoms," a Department of Health and Human Services (DHHS) statement read.
In the past month in regional Victoria, positive readings for COVID-19 wastewater fragments have been detected in Bacchus Marsh, Benalla, Bendigo, Black Rock, Healesville, Koorlong, Lakes Entrance, Somers and Wangaratta.
No known cases in Benalla, Healesville or Wangaratta have been found.
Exposure sites swelled above 450 overnight but none had been listed in regional areas.
Saturday's cases were detected from 33,675 tests, with 29,490 vaccine doses administered across state-run sites.
More to come.
