Coping with formalin banning in pathology: under vacuum long-term tissue storage with no added formalin.

Formalin is toxic and has recently been classified as carcinogenic leading to a proposed European formalin ban. But, the pathology use of formalin has however been completely overlooked, and this is proving to be a relevant issue, as no alternative, reliable, tissue fixative is available. Various systems have been proposed to reduce formalin use and exposure; long-term storage and disposal of formalin is also a problem. With this in mind, under vacuum sealing (UVS) systems have been proposed for transportation/storage, however, for how long tissue retains its characteristics (morphological and molecular) is unknown. This study aims to compare histology specimens stored by formalin immersion (FI) and specimens stored after fixation with UVS technique with no additional formalin, at different time periods. Twenty tissue samples (10FI; 10UVS) were stored for different time periods (15 days, 1-2-3-6-12 months) for a total of 120 samples, compared with regard to their morphology, histochemistry, immunoreactivity (24 specific antibodies) and DNA status. All samples showed well-preserved morphology and overlapping staining quality. A significant reduction in immunoreactivity was however identified in the various time periods, particularly for heat pre-treated nuclear antigens, and this commenced earlier (1 month) for FI. UVS storage showed higher DNA content than FI but slightly poorer DNA integrity. These results add important knowledge to the use of UVS in daily practice, as long-term storage of pre-fixed tissue in UVS is not detrimental to the quality of tissue while having the boon of using very little formalin with less operator exposure and lower disposal costs.

Collagenase production in an antarctic strain of Arthrobotrys tortor Jarowaja.

This paper describes the results of a comparative screening between the nematophagous Antarctic fungus Arthrobotrys tortor and other species of that genus for the production of extracellular collagenases. The nematode species used in this study was Caenorhabditis elegans, feeding on Escherichia coli cultures. Determination of collagenase activity was made using insoluble collagen from bovine Achilles tendon and determining the amount of solubilized hydroxyproline produced. The results show that the total amount of collagenase produced by the Antarctic strain of A. tortor was about threefold higher than that observed for the other species. In the Antarctic strain, collagenase was shown to be a constitutive enzyme.