In order to evaluate the potential risk of Alicyclobacillus contamination in Shaanxi kiwi fruit production line, six strains of Alicyclobacillus isolated from the production line were studied. The contaminant production capacity of six strains and four corresponding standard strains was evaluated, including detection of contaminant production using GC—MS and heat resistance through drawing thermal death curves, and its biodiversity was analyzed, including analysis of carbon source spectrum by API tests and detection of its DNA hybridizations from renaturation rates. The results showed that no 2,6-dibromophenol and 2,6-dichlorophenol were detected, and the guaiacol production of six isolated strains was between 10.56μg/L and 34.74μg/L which was significantly higher than the common sensory detection threshold values in juices, D90℃ values of strains were 27.1~34.2 min and the z values were 12.14~13.66℃ in kiwi fruit juice, with the heat resistance of the three strains of A. acidoterrestris generally being higher than the data of former reports. There were 7~22 different results of API tests between the isolated strains and the data from published articles or their corresponding standard strains. The hybridizations between the isolated strains and their corresponding standard strains were between 74.75% and 104.79%. These results suggested that the isolated strains possessed potential contaminant production capacity and biodiversity to a certain extent which needed to be controlled.