132 Acta argiculturae Slovenica, Supplement 5, 132–137, Ljubljana 2016 24th Int. Symp. “Animal Science Days”, Ptuj, Slovenia, Sept. 21st−23rd, 2016. COBISS: 1.08 Agris category code: Q02 EFFECTS OF SALTING DURATION AND BOAR TAINT LEVEL ON QUALITY OF DRY-CURED HAMS Tadej KALTNEKAR 1, 2, Martin ŠKRLEP 3, Nina BATOREK LUKAČ 3, Urška TOMAŽIN 3, Maja PREVOLNIK POVŠE 3, 4, Etienne LABUSSIERE 5, Lea DEMŠAR 6, Marjeta ČANDEK- POTOKAR 3, 4, 7 Effects of salting duration and boar taint level on quality of dry-cured hams 1 Pršutarna Lokev na Krasu, Lokev 9, 6219 Lokev, Slovenia, e-mail: prsutarna.lokev@gmail.si 2 The study issues from the graduation thesis of Tadej Kaltnekar (supervisor prof. dr. Lea Demšar). 3 Agricultural Institute of Slovenia, Hacquetova ulica 17, 1000 Ljubljana, Slovenia, e-mail: martin.skrlep@kis.si, nina.batorek@kis.si, urska.tomazin@kis.si 4 University of Maribor, Faculty of Agriculture and Life Sciences, Pivola 10, 2311 Hoče, Slovenia, e-mail: maja.prevolnik@um.si 5 INRA, UMR 1348 Pegase, 35590 Saint-Gilles, France and Agrocampus-Ouest, UMR 1348 Pegase, 35000 Rennes, France, e-mail: etienne.labussiere@rennes.inra.fr 6 University of Ljubljana, Biotechnical Faculty, Food Science and Technology Dpt., Jamnikarjeva 101, 1000 Ljubljana, e-mail: lea.demsar@bf.uni-lj.si 7 Corresponding author: e-mail: meta.candek-potokar@kis.si ABSTRACT Meat from entire males (EM) may take an important market share in the EU in case surgical castration is aban- doned. The aim of the present study was to assess the effect of salting regime and boar taint level on dry-cured ham quality. Standard (18 days, HS) and shortened salting (6 days, LS) was applied to 16 hams from 8 EM, being also evalu- ated according to boar taint; low (LBT) or high (HBT). Compared to standard procedure, shorter salting resulted in 40  % lower salt content, higher proteolysis, higher aw, softer texture, accompanied by lower sensorial saltiness and sourness and higher meat colour intensity, sweetness, bitterness, pastiness and off-flavour perception. HBT hams were more proteolysed, with softer texture, higher perception of bitterness, pastiness and juiciness with more pronounced off- flavours and poorer typical cured odour than LBT hams. A significant interaction between boar taint level and salting on perceived off flavours denotes that either salt covers off-flavours or there are additive effects of boar taint substances and proteolysis products when salt content is low. Key words: pigs, entire male pigs, meat, dry-cured ham quality, boar taint, salting regime 1 INTRODUCTION Traditional dry-cured ham Kraški pršut is the most appreciated dry-cured meat product among Slovenian consumers (Čandek-Potokar and Arh, 2004). Appropri- ate raw material quality is essential for high quality prod- uct. For Kraški pršut, the origin of raw material is not prescribed, thus the producers are strongly bound to the supply of green hams from the European markets. Due to the initiative to stop surgical castration of piglets, it is ex- pected that rearing of entire males (EM) will take signifi- cant share in European pig production. Besides accumu- lation of malodorous boar taint substances (skatole and androstenone) in fat tissue, EM deposit less fat (subcuta- neous as well as intramuscular – IMF) and exhibit lower water holding capacity (WHC) of meat (Batorek et al., 2012; Pauly et al., 2012) which may reduce dry-cured ham quality. Namely, poor WHC causes lower season- ing yields and higher salt uptake, resulting in dry, firm and salty product with poor aroma (Čandek-Potokar and Škrlep, 2012). Proper amount of fat prevents excessive water loss and serves as a barrier for salt uptake, with IMF improving the development of typical aromatic and tex- tural properties. Furthermore, EM exhibit higher protein deposition capacity, which could affect the proteolytic potential of EM meat and induce changes associated to the process of proteolysis during the lengthy dry-curing process (Toldra, 2002). As the information about the use of EM for dry-cured ham production is scarce, the aim of this study was to investigate the influences of such raw material on ham processing and final quality considering different salting regimes and boar taint levels. Acta agriculturae Slovenica, Supplement 5 – 2016 133 EFFECTS OF SALTING DURATION AND BOAR TAINT LEVEL ON QUALITY OF DRY-CURED HAMS Salting Boar taint p-value RMSE HS LS LBT HBT Salting Boar taint Salting × boar taint Green ham traits Ham weight (kg) 12.9 12.7 12.9 12.6 0.768 0.688 0.948 1.5 pH SM 5.49 5.47 5.50 5.46 0.664 0.305 0.492 0.07 Fat thickness (mm) 16 13 16 13 0.235 0.108 0.973 4 Processing losses (%) Salting 6 days 2.0 1.9 1.8 2.1 0.856 0.182 0.871 0.5 Salting 18 days 3.9 - - - - - - - Resting 20.1 20.0 19.2 20.9 0.990 0.207 0.976 2.5 Drying 27.6 27.2 26.4 28.3 0.809 0.234 0.944 3.1 Ripening 36.6 36.6 35.2 38.0 0.984 0.237 0.902 4.6 Table 1: Raw material traits and processing losses (LS means) according to salting duration and boar taint HS – salting 18 days; LS – salting 6 days; HBT – androstenone content in subcutaneous fat > 0.78 µg/g; LBT – androstenone content in subcutaneous fat < 0.78 µg/g; SM – semimembranosus muscle; RMSE – root-mean-square error Salting Boar taint p-value RMSE HS LS LBT HBT Salting Boar taint Salting × boar taint SM muscle Salt (g/kg) 67.0 37.8 50.8 54.0 0.000 0.472 0.948 8.7 Dry matter (g/kg) 489.3 471.1 470.5 489.9 0.214 0.186 0.894 27.6 IMF (g/kg) 44.1 39.6 40.8 43.0 0.270 0.574 0.900 7.8 NPN (g/kg) 13.2 15.2 13.6 14.8 0.002 0.045 0.332 1.1 Proteolysis index (%) 22.1 25.3 23.1 24.3 0.003 0.183 0.283 1.8 aw 0.894 0.940 0.921 0.913 0.0001 0.397 0.919 0.017 BF muscle Salt (g/kg) 77.4 45.8 58.9 64.4 0.000 0.217 0.813 8.4 Dry matter (g/kg) 405.5 389.4 390.5 404.4 0.139 0.198 0.895 20.3 IMF (g/kg) 29.1 30.3 29.9 29.5 0.634 0.874 0.874 4.6 NPN (g/kg) 13.7 15.6 13.8 15.5 0.034 0.048 0.659 1.5 Proteolysis index (%) 29.8 33.5 30.0 33.3 0.024 0.042 0.666 2.9 aw 0.895 0.937 0.920 0.912 0.000 0.227 0.815 0.014 Subcutaneous fat Androstenone (µg/g) 0.97 1.01 0.65 1.32 0.846 0.005 0.885 0.39 Skatole (µg/g) 0.17 0.15 0.08 0.24 0.811 0.013 0.838 0.11 Table 2: Effect of salting duration and boar taint on chemical traits (LS means) of dry-cured hams HS – salting 18 days; LS – salting 6 days; HBT – androstenone in subcutaneous fat > 0.78 µg/g; LBT – androstenone in subcutaneous fat < 0.78 µg/g; SM – semimembranosus muscle; BF – biceps femoris muscle; IMF – intramuscular fat; NPN – non-protein nitrogen; aw – water activity; RMSE – root-mean-square error Acta agriculturae Slovenica, Supplement 5 – 2016134 T. KALTNEKAR et al. 2 MATERIALS AND METHODS Green hams (n = 16; 12.8 ± 1.4 kg) from 8 crossbred boars fed commercial feed mixture were processed in a commercial facility according to the rules of consor- tium Kraški pršut. Measurements of green ham weight, subcutaneous fat thickness and pH in semimembranosus (SM) muscle were recorded prior to salting at 2–4 °C for 6 days (LS – shortened salting, right hams) or 18 days (HS – standard salting regime, left hams). After the salt- ing, the hams were washed and left to rest at 4–6 °C and 70–85 % relative humidity (RH) for 89 and 77 days for LS and HS, respectively. Following resting, the hams were left to dry and ripen (14–20 °C, 60–80 % RH) until reach- ing the final processing age of 68 weeks. In order to mon- itor processing losses, ham weight was recorded at the end of each phase. Boar taint substances androstenone and skatole were determined by HPLC (as described by Batorek et al., 2012) in samples of subcutaneous fat taken at the level of the last rib at slaughter and from the cen- tral part of the ham at the end of the ripening. Chemical analysis (salt, total nitrogen content, non-protein nitro- gen – NPN, proteolysis index – IP, IMF) and rheologi- cal measurements (force decay coefficient, texture profile analysis) were performed in SM and biceps femoris (BF) muscle samples as described in Pugliese et al. (2015). Sensory analysis (quantitative descriptive analysis) was conducted with 11 trained panellists evaluating sensory descriptors on entire ham slice (marbling, colour homo- geneity and intensity, typical cured odour), subcutaneous fat (whiteness, rancidity, sweetness, off-flavours) and sep- arately SM and BF muscles (saltiness, acidity, sweetness, bitterness, off-flavours, solubility, juiciness, pastiness). Data was statistically analysed using GLM procedure of the SAS/STAT module (SAS Institute, Cary, NC, USA). The median of androstenone concentration (0.78  µg/g liquid fat) measured in subcutaneous fat of dry-cured hams was used to classify the hams into low (LBT; an- drostenone < 0.78 µg/g) and high boar taint (HBT; an- drostenone > 0.78 µg/g). This benchmark agrees with the level considered as threshold for sensorial perception i.e. 0.5−1.0 µg/g fat (Walstra et al., 1999). Salting time, boar taint level, and their interaction were included as fixed effects. Differences between groups were considered sig- nificant at p < 0.05. 3 RESULTS AND DISCUSSION Green ham traits were recorded due to their im- portance for dehydration, salt intake and biochemical changes. No effect (p > 0.10) of salting regime or boar Salting Boar taint p-value RMSE HS LS LBT HBT Salting Boar taint Salting × boar taint SM muscle Force decay coefficient 0.62 0.67 0.63 0.65 0.005 0.148 0.980 0.03 Hardness (N) 90.1 54.3 69.9 74.5 0.006 0.671 0.433 21.4 Cohesiveness 0.49 0.43 0.49 0.43 0.111 0.119 0.597 0.07 Gumminess (N) 54.2 24.0 35.2 43.0 0.008 0.426 0.143 19.0 Springiness (mm) 3.5 3.1 3.4 3.2 0.190 0.377 0.935 0.5 Chewiness (N) 191.4 77.7 128.3 140.8 0.009 0.737 0.226 72.7 Adhesiveness (N*mm) −2.4 −3.3 −2.1 −3.5 0.027 0.003 0.267 0.8 BF muscle Force decay coefficient 0.68 0.71 0.69 0.70 0.083 0.890 0.980 0.04 Hardness (N) 42.3 25.8 38.7 29.3 0.004 0.067 0.931 9.3 Cohesiveness 0.51 0.41 0.52 0.40 0.029 0.016 0.841 0.08 Gumminess (N) 23.1 11.2 21.4 12.9 0.009 0.047 0.716 7.7 Springiness (mm) 3.6 3.6 3.6 3.6 0.892 0.964 0.102 0.7 Chewiness (N) 87.1 38.5 81.7 43.9 0.016 0.049 0.369 34.6 Adhesiveness (N*mm) −1.0 −1.5 −0.9 −1.5 0.188 0.127 0.299 0.8 Table 3: Effect of salting duration and boar taint on rheological traits (LS means) of dry-cured hams HS – salting 18 days; LS – salting 6 days; HBT – androstenone in subcutaneous fat > 0.78 µg/g; LBT – androstenone in subcutaneous fat < 0.78 µg/g; SM – semimembranosus muscle; BF – biceps femoris muscle; RMSE – root-mean-square error Acta agriculturae Slovenica, Supplement 5 – 2016 135 EFFECTS OF SALTING DURATION AND BOAR TAINT LEVEL ON QUALITY OF DRY-CURED HAMS taint level was observed confirming that groups were equilibrated in terms of raw material properties (Table 1). Reducing salting from 18 to 6 days led to 40 % salt reduction (in BF and SM; Table 2), and resulted in higher aw and proteolysis index (p < 0.05) in LS than HS hams, which corroborates inversely proportional relationship between salt content and protein breakdown (Martín et al., 1998). Proteolysis index in BF muscle of LS hams exceeded (in average) the level of 30 %, which has been associated with flavour and texture defects (Virgili et al., 1995). Substantial salt reduction can be problematic also for product safety. LS hams in our study were microbio- logically tested and were proven safe. However, average aw (SM and BF) was slightly above 0.93, a prescribed limit for dry meat products (UL RS, 59/2012). Salt reduc- tion is positive for lower risk of cardiovascular diseases, but stability of such products may be compromised as high aw facilitates microbial growth (Bem et al., 2003) and thus lowers product shelf life. Notably higher NPN (both muscles) and IP (BF) (p < 0.05) observed in HBT than LBT can be explained with androstenone which re- flects higher anabolic potential, higher protein turnover Salting Boar taint p-value RMSE HS LS LBT HBT Salting Boar taint Salting × boar taint Entire slice Meat colour uniformity 6.3 6.4 6.3 6.4 0.524 0.647 0.247 0.4 Meat colour intensity 5.0 5.5 5.2 5.3 0.032 0.667 0.205 0.4 Marbling 2.2 2.5 2.4 2.3 0.400 0.783 0.974 0.6 Typical cured odour 5.4 5.2 5.6 5.0 0.384 0.038 0.234 0.5 Subcutaneous fat Fat whiteness 5.8 5.3 5.5 5.6 0.298 0.929 0.867 1.0 Fat sweetness 4.1 3.5 3.8 3.7 0.004 0.531 0.808 0.3 Fat off-flavour 1.0 1.3 0.9 1.4 0.254 0.081 0.330 0.4 Fat rancidity 1.6 1.7 1.7 1.6 0.642 0.974 0.822 0.5 SM muscle Bitterness 0.6 1.3 0.8 1.1 0.001 0.053 0.114 0.3 Sourness 1.8 1.5 1.6 1.6 0.030 0.822 0.475 0.3 Pastiness 1.1 2.1 1.0 2.2 0.032 0.017 0.143 0.8 Sweetness 0.6 1.3 0.9 1.0 <0.0001 0.662 0.226 0.2 Saltiness 6.0 4.0 5.2 4.8 <0.0001 0.227 0.299 0.6 Juiciness 3.8 4.4 4.3 4.0 0.046 0.357 0.205 0.5 Solubility 4.9 5.2 4.9 5.2 0.251 0.109 0.182 0.4 Off-flavour 0.7 1.7 0.9 1.5 0.0001 0.006 0.071 0.3 BF muscle Bitterness 0.7 1.3 0.8 1.2 0.001 0.014 0.227 0.3 Sourness 2.3 2.0 2.1 2.2 0.140 0.438 0.890 0.3 Pastiness 1.5 2.8 1.4 2.9 0.039 0.019 0.504 1.0 Sweetness 0.6 1.2 0.9 1.0 <0.0001 0.138 0.180 0.1 Saltiness 6.8 4.8 5.9 5.7 <0.0001 0.421 0.655 0.5 Juiciness 5.2 5.4 5.3 5.3 0.188 0.856 0.051 0.4 Solubility 5.2 5.2 5.0 5.5 0.952 0.019 0.177 0.4 Off-flavour 0.9 1.9 1.0 1.7 0.0001 0.002 0.014 0.3 Table 4: Effect of salting duration and boar taint on sensory traits (LS means) of dry-cured hams HS – salting 18 days; LS – salting 6 days; HBT – androstenone content in subcutaneous fat > 0.78 µg/g; LBT – androstenone content in subcutaneous fat < 0.78 µg/g; SM – semimembranosus muscle; BF – biceps femoris muscle; RMSE – root-mean-square error Acta agriculturae Slovenica, Supplement 5 – 2016136 T. KALTNEKAR et al. and thus also increased proteolytic activity (Claus et al. 1994). There was no effect of salting on the level of boar taint compounds. LBT was in average below and HBT above benchmark considered a threshold for sensory perception (Walstra et al., 1999). Salting duration affected majority of evaluated rhe- ological parameters (Table 3). Compared to HS, hams of the LS group had softer, less gummy and chewy texture (p < 0.05) with higher force decay coefficient (p < 0.10) in both investigated muscles, along with reduced cohe- siveness (p < 0.05) and increased adhesiveness (p < 0.05) in BF and SM, respectively. Noted impacts are in line with chemical traits namely the effect of low salt con- tent and higher proteolysis and/or aw on increased pro- tein breakdown and consequently softer texture (Ruiz- Ramírez et al., 2006). Considering the effect of boar taint, HBT had higher adhesiveness (p < 0.05) in SM and lower hardness (p < 0.10), cohesiveness, gumminess and chewiness (p < 0.05) than LBT hams. As in the case of salting regime, the effect can be related to differences in proteolysis observed between HBT and LBT groups. Salting duration showed notable effect (p < 0.05) on a good number of investigated sensory traits (Table 4). Compared to standard procedure, shorter salting was as- sociated with higher colour intensity (entire ham slice), lower saltiness, higher bitterness, pastiness, sweetness (BF and SM) and higher juiciness (SM) along with lower fat sweetness. In addition, higher degree of off-flavours was perceived in both LS ham muscles. While traits like saltiness and sourness perception can be related to the level of salt i.e. chloride ions (Buscailhon et al., 1995), other effects are most probably related to the differences in proteolysis. More intensive colour of LS hams could be explained by the higher level of stable red pigment, related to the degree of proteolytic degradation (Grossi et al., 2014). Other traits like increased pastiness, higher bitterness and off-flavour presence are indicating exces- sive protein degradation, resulting in their structural breakdown as well as numerous free amino acids and short peptides, which can negatively affect sensory per- ception (Virgili and Schivazappa, 2002). Regarding the association of boar taint level with sensory traits, both muscles of HBT group were evalu- ated as more pasty and bitter with higher degree of off- flavours in addition to higher BF solubility (p < 0.05) and off-flavour presence in ham subcutaneous fat (p < 0.10). Despite lipophilic nature of boar taint substances, the dif- ference in off-flavour was much more evident in muscle (with low IMF) than fat tissue indicating that increased protein breakdown could be an important source of the off-flavours in HBT hams. Although a negative effect of boar taint substances on off-flavour perception could be anticipated, the effect of increased proteolysis may play a significant role in this case. An interaction between salting and boar taint (Ta- ble 5) was observed for off-flavours in BF (p < 0.05) and SM (p < 0.10). In HS hams slightly higher score of off- flavour perception in HBT than LBT was not significant, whereas in LS hams HBT had notably higher off-flavour score than LBT in muscle (p < 0.05) and in fat (p > 0.10) tissue. This indicates that either salt covers foreign fla- vours when present in higher amounts (Breslin and Beauchamp, 1997) or there are additive effects of boar taint and proteolysis on products when the amount of salt is low. 4 CONCLUSIONS Substantial reduction of salting duration consider- ably reduced salt content and affected other dry-cured ham properties, but not always in a favourable manner. Interestingly, higher boar taint level was associated with increased proteolytic activity, pointing out an additional issue for dry-cured hams, processed from EM meat, es- pecially in combination with lower salt content. Salting HS LS p-value RMSE Boar taint LBT HBT LBT HBT Salting Boar taint Salting × boar taint Off-flavour BF 0.8a 1.0a 1.3a 2.5b 0.0001 0.002 0.014 0.3 SM 0.6a 0.8a 1.2a 2.1b 0.001 0.006 0.071 0.3 Fat 0.9 1.1 1.0 1.6 0.254 0.081 0.330 0.4 Table 5: Interacting effect of salting and boar taint on off-flavours in different dry-cured ham tissues HS – salting 18 days; LS – salting 6 days; HBT – androstenone content in subcutaneous fat > 0.78 µg/g; LBT – androstenone content in subcutaneous fat < 0.78 µg/g; SM – semimembranosus muscle; BF – biceps femoris muscle; RMSE – root-mean-square error; a,b Means within a row with different superscripts differ at p < 0.05 Acta agriculturae Slovenica, Supplement 5 – 2016 137 EFFECTS OF SALTING DURATION AND BOAR TAINT LEVEL ON QUALITY OF DRY-CURED HAMS 5 ACKNOWLEDGMENTS The authors acknowledge financial support of the Slovenian Research Agency and Ministry of Agriculture, Food and Forestry (grants P4-0133, L4-5521 and V4- 1417). 6 REFERENCES Batorek, N., Škrlep, M., Prunier, A., Louveau, I., Noblet, J., Bon- neau, M., Čandek-Potokar M. (2012). Effect of feed restric- tion on hormones, performance, carcass traits, and meat quality in immunocastrated pigs. Journal of Animal Science, 90, 4593–4603. Bem, Z., Adamič, J., Žlender, B., Smole Možina, S., Gašperlin L. (2003). Mikrobiologija živil živalskega izvora. Ljubljana: Biotehniška fakulteta, Oddelek za živilstvo. Breslin, P.A.S., Beauchamp, G.K. (1997). Salt enhances flavour by suppressing bitterness. Nature, 387: 563. Buscailhon, S., Touraille, C., Girard, J.P., Monin, G. (1995). Re- lationships between muscle tissue characteristics and sen- sory qualities of dry-cured ham. Journal of Muscle Foods, 6, 9−22. Claus, R., Weiler, U., Herzog, A. (1994). Physiological aspects of androstenone and skatole formation in the boar – a review with experimental data. Meat Science, 38, 289–305. Čandek-Potokar, M., Arh, M. (2004). Evaluating market pros- pects for Prekmurje dry ham in relation to consumption characteristics of drymeat products in Slovenia. In A. Audi- ot, F. Casabianca, & G. Monin (Eds.), Proceedings of the 5th International Symposium on the Mediterranean Pig, Tarbes (France), 16–19 November 2004 (Options Méditerranéennes, No. 76) (pp. 327–332). Zaragoza, Spain: Instituto Agro- nomico Mediterraneo de Zaragoza. Čandek-Potokar, M., Škrlep, M. (2012). Factors in pig produc- tion that impact the quality of dry-cured ham: A review. Animal, 6, 327–338. Grossi, A.B., do Nascimento, E.S.P., Cardoso, D.R., Skibsted, L.H. (2014). Proteolysis involvment in zinc-protoporphy- rin IX formation during Parma ham formation. Food Re- search International, 56, 252–259. Martín, L., Cordoba, J.J., Antequera, T., Timón, M., Ventanas, J. (1998). Effects of salt and temperature on proteolysis dur- ing ripening of Iberian ham. Meat Science, 49, 145−153. Pauly, K., Luginbühl, W., Ampuero, S., Bee, G. (2012). Expected effects on carcass and pork quality when surgical castration is omitted. Meat Science, 92, 858–862. Pravilnik o kakovosti mesnih izdelkov. (2012). Ur. L. RS, št. 59/2012. Pugliese, C., Sirtori, F., Škrlep, M., Piasentier, E., Calamai, L., Franci, O., Čandek-Potokar, M. (2015). The effect of rip- ening time on the chemical, textural, volatile and sensorial traits of biceps femoris and semimembranosus muscles of the Slovenian dry-cured ham Kraški pršut. Meat Science, 100, 58−68. Ruiz-Ramírez J., Arnau J., Serra X., Gou P. (2005). Relationship between water content, NaCl content, pH and texture pa- rameters in dry-cured muscles. Meat Science, 70: 579−587. Toldra F. 2002. Dry-cured meat products. Trumbull: Food and Nutrition Press, Inc. Virgili R., Parolari, G., Schivazappa, C., Soresi-Bordini, M., Bor- ri, M. (1995). Sensory and texture quality of dry-cured ham as affected by endogenous cathepsin B activity and muscle composition. Journal of Food Science, 60, 1183−1186. Virgili, R., Schivazappa, C. (2002). Muscle traits for long ma- tured dried meats. Meat Science, 62, 331−343. Walstra, P., Claudi-Magnussen, C., Chevillon, P., von Seth, G., Diestre, A., Matthew,s K.R., Homer, D.B. et al. (1999). An international study on the importance of androstenone and skatole for boar taint: levels of androstenone and skatole by country and season. Livestock Production Science, 62, 15−28.