Reduction of Clostridium sporogenes spore outgrowth in natural sausage casings using nisin.

Preservation of natural sausage casings using dry salt or saturated brine is regarded as sufficient to inactivate vegetative pathogenic non-spore-forming bacteria present on the casings. Although the outgrowth of bacterial spores is prevented by salt or saturated brine preservation, these spores will remain present and develop into vegetative cells when conditions are more favourable. To prevent subsequent outgrowth additional preservation measures should be implemented. In the experiments described the use of nisin was evaluated to reduce outgrowth of spores in desalinated casings. The bacteriocin nisin was chosen because of its known efficacy against spore-forming bacteria and their spores in various foodstuffs. Clostridium spore suspensions (Clostridium sporogenes, ATCC 3584) were used in two concentrations to inoculate three nisin concentrations (10, 50, 100 µg/mL) in water containing gamma-irradiated casings. Additionally, the binding of nisin to casings, using (14)C-labeled nisin Z and subsequent availability of nisin were evaluated. Results demonstrate that nisin is partly reversibly bound to casings and can reduce the outgrowth of Clostridium spores in the model used by approximately 1 log(10) (90%). However, the biological relevance of these results needs to be determined further by conducting industrial trials before any recommendation can be made on the practical implementation of nisin in the preservation of natural sausage casings.

Phosphate analysis of natural sausage casings preserved in brines with phosphate additives as inactivating agent - Method validation.

Certain phosphates have been identified as suitable additives for the improvement of the microbial and mechanical properties of processed natural sausage casings. When mixed with NaCl (sodium chloride) and used under specific treatment and storage conditions, these phosphates are found to prevent the spread of foot-and-mouth disease and classical swine fever via treated casings. The commercially available Quantichrom™ phosphate assay kit has been evaluated as to whether it can serve as a reliable and low-tech method for routine analysis of casings treated with phosphate. The outcome of this study indicates that this particular assay kit has sufficient sensitivity to qualitatively determine the presence of phosphate in treated casings without interference of naturally occurring phosphate in salt used for brines in which casings are preserved.

Inactivation of classical swine fever virus in porcine casing preserved in salt.

Pig intestines used for the production of natural sausage casings may carry classical swine fever (CSF) virus. Feeding pigs with human food waste that contains pig casings may then spread the virus to CSF-free animals. Casings derived from a pig experimentally infected with CSF by dosing with 10(6) tissue culture infectious doses (TCID50) of the highly virulent CSF virus strain "Koslov", were treated with phosphate supplemented or citrate supplemented NaCl, instead of with NaCl alone, which is the standard preservation treatment for casings. Treated casings were stored for 30 days at either 4 degrees C or 20 degrees C. After storage the casings were fed to 16 susceptible pigs. CSF infection was confirmed in the four animals that had been fed casings treated with citrate supplemented salt and stored at 4 degrees C. All other animals remained healthy. It is therefore possible to avoid the inadvertent spread of CSF virus via porcine sausage casings by treating casings with phosphate supplemented salt and storing them for 30 days at temperatures over 4 degrees C.

Removal of foot-and-mouth disease virus infectivity in salted natural casings by minor adaptation of standardized industrial procedures.

Intestines are used for the production of natural casings as edible sausage containers. Derived from animals (pigs and sheep) experimentally infected with FMDV (initial dosage 10(7.3) PFU/ml, strain O(1Kaufbeuren)), these natural casings were treated with sodium chloride or a phosphate salts/sodium chloride mixture and the residual FMDV titres measured. After storage at about 20 degrees C, no remaining infectivity was found after either treatment, whereas casings stored at 4 degrees C still contained infectivity. Storage of salted casings at about 20 degrees C for 30 days is already part of the Standard Operating Procedures (included in HACCP) of the international casing industry and can therefore be considered as a protective measure for the international trade in natural casings.

Antimicrobial properties of salt (NaCl) used for the preservation of natural casings.

The antimicrobial properties of salt (NaCl) used for the preservation of natural casings were studied by investigating the survival of six bacterial species in natural casings at different water activity (aw) levels. Individual sheep casings were inoculated with ca. 10(5) colony-forming units (cfu) g(-1) of Escherichia coli, Salmonella typhimurium, Listeria monocytogenes, Staphylococcus aureus, Clostridium perfringens and 10(2)cfu g(-1) of E. coli O157:H7. The casings were stored at 20+/-1.5 degrees C in different brines and dry salt, giving aw-levels of 0.90 aw, 0.87aw, 0.85 aw, 0.83 aw and 0.75 aw. Samples were taken at day 1, 3, 6, 8, 13, 20, 27 and 30 after inoculation and the number of bacteria present was determined. Based on survival curves, death rates (day(-1)) were calculated to quantify the reduction in log10 cfu g(-1) per day. The influence of aw on death rates was higher for Gram-negative bacteria than for Gram-positive bacteria. The death rates were overall higher for Gram-negatives than for Gram-positives. No clear reduction in the survival of C. perfringens in relation to any aw level was observed in this study. These results indicate that the antimicrobial properties of salt used for the preservation of natural casings are sufficient to reduce the bacterial contamination (except for Clostridium spores) well below acceptable levels at a water activity level of 0.85 or lower during a 30-day storage period.