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Changes in the physico-chemical attributes through processing of salami made from blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) in comparison to pork
Accepted Manuscript Changes in the physico-chemical attributes through processing of salami made from blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) in comparison to pork
Chido Chakanya, Elodie Arnaud, Voster Muchenje, Louwrens C. Hoffman PII: DOI: Reference:
S0309-1740(18)30409-1 doi:10.1016/j.meatsci.2018.07.034 MESC 7648
To appear in:
Meat Science
Received date: Revised date: Accepted date:
16 April 2018 22 June 2018 30 July 2018
Please cite this article as: Chido Chakanya, Elodie Arnaud, Voster Muchenje, Louwrens C. Hoffman , Changes in the physico-chemical attributes through processing of salami made from blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) in comparison to pork. Mesc (2018), doi:10.1016/j.meatsci.2018.07.034
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ACCEPTED MANUSCRIPT Changes in the physico-chemical attributes through processing of salami made from blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) in
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comparison to pork
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Chido Chakanya1, Elodie Arnaud2,3,4, Voster Muchenje1 and Louwrens C. Hoffman2,* Department of Livestock and Pasture Science, University of Fort Hare, Private Bag X 1314,
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Alice 5700, South Africa
Department of Animal Sciences, Stellenbosch University, Private Bag X1 Matieland,
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Stellenbosch 7602, South Africa
CIRAD, UMR QualiSud, 7602 Stellenbosch, South Africa
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Qualisud, Univ Montpellier, CIRAD, Montpellier SupAgro, Université d'Avignon, Université
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de La Réunion, Montpellier, France
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*Corresponding author: [email protected]
ACCEPTED MANUSCRIPT Abstract Drying kinetics and changes in proximate composition, pH, salt content, water activity (aw) and lipid oxidation through processing of salami made using five different game meat species were evaluated and compared to pork. Eight batches of salami from each species were made and
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sampled for analysis throughout processing. Processing time was a significant factor on all
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measured attributes whilst species affected (P≤0.05) pH and moisture but not drying kinetics.
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Black wildebeest meat exhibited higher (P≤0.05) pH than pork and other game meat (6.30 vs 5.63-5.80), which translated to higher (P≤0.05) salami pH throughout and at the end of
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processing (5.77). Final pH of all other salami ranged from 5.01 to 5.28, aw ranged from 0.88 to
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0.92. TBARS remained lower than 1 mg MDA equivalent/kg. The study suggests that salami from these game species, excluding black wildebeest, can be produced using the same processing
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parameters as conventional pork salami and obtaining similar physico-chemical attributes.
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Key words: fermented sausages; game meat; lipid oxidation; TBARS; drying kinetics
ACCEPTED MANUSCRIPT 1. Introduction The favorable nutritional and physico-chemical profile of meat from game species has been shown over the years (Hoffman & Cawthorn, 2013). Furthermore, an increase in the production of meat from game species has been noted in some regions, giving opportunity for the expansion
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of the game meat industry (Van Schalkwyk, McMillin, Witthuhn, & Hoffman, 2010; Ljung,
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Riley, & Ericsson, 2015). In South Africa, most game meat is a by-product of trophy hunting/and or culling activities (McCrindle, Siegmund-Schultze, Heeb, Zárate, & Ramrajh, 2013). Game
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meat is commonly used for the production of biltong made from salted and dried meat strips (Jones, Arnaud, Gouws, & Hoffman, 2017) and fermented sausage (salami) (Van Schalkwyk et
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al., 2011; Van de Merwe, Saayman, & Rossouw, 2015) as it is sometimes perceived as being
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tough and hard to cook (Hoffman, Muller, Schutte, & Crafford, 2004).
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Fermented sausages are popularly produced using pork meat and fat (Ruiz, 2007; Yilmaz, Velioglu, Lachowicz, & Joanna, 2009). Their production can be separated into three distinct
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phases; formulation, fermentation and drying/ripening. Formulation entails the preparation of
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seasoned, minced raw meat and fat for encasing whilst fermentation is crucial for the production of lactic acid which lowers pH and drying/ripening ensures the lowering of water activity (aw)
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through moisture loss (Ordóñez, Hierro, Bruna, & de la Hoz, 1999). The shelf stability of fermented sausages is based on the combined hurdle techniques of lowering pH and aw in order to create an environment that impedes growth of spoilage bacteria (Toldrá, 2002). Generally, the relationship between aw and pH of fermented products should be inversely proportional to ensure shelf life stability, i.e the higher the final pH, the lower the aw should be (Zukal & Incze, 2010). Meat products are considered shelf stable when they have a pH lower than 5 or aw lower than 0.91 or a combination of pH lower than 5.2 and aw below 0.95 (Leistner & Rodel, 1975).
ACCEPTED MANUSCRIPT However, there is still some debate about the precise pH and aw as noted by Abunyewa, Laing, Hugo, & Viljoen (2000). It is known that the characteristics of the raw material used impacts on the quality of the end product in sausage fermentation (Ruiz, 2007). Game meat is known to have lower intra muscular
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fat, higher amounts of polyunsaturated fatty acids as well as higher heme iron and myoglobin
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contents (Hoffman & Wiklund, 2006) compared to pork. These characteristics may have a significant bearing on its fermentation quality. Production of fermented sausages using game
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meat is relatively unexplored especially on wildlife species found in Southern Africa. Studies in which fermented sausages were produced using game meat and pork as a fat source have focused
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on wild boar and deer (Paulsen, Vali, & Bauer, 2011; Cenci-Goga et al., 2012; Utrilla, García
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Ruiz, & Soriano, 2014; Maskimovic et al., 2018). Todorov et al. (2007) and Van Schalkwyk et al. (2011) looked at the physico-chemical, microbial and sensory quality of fermented sausages
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made using springbok (Antidorcas marsupialis), gemsbok (Oryx gazella), kudu (Tragelaphus
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strepsiceros), zebra (Equus burchelli) and blesbok (Damaliscus pygargus phillipsi). However, little is known on the physico-chemical changes that occur during their processing. These
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changes are critical in developing guidelines for the production of fermented sausages for
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individual species with consistently high quality. Therefore this study aimed at characterizing the changes that occur during production of salami made using species commonly harvested in South Africa ie. blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) and comparing them to pork salami.
ACCEPTED MANUSCRIPT 2. Materials and methods 2.1 Raw meat Meat was sourced from the skeletal muscles and trimmings after removal of major muscles (semitendinosus, biceps femoris, infra and supra spinatus, semimembranosus, longissimus
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thoracis et lamborum) of blesbok, eland, fallow deer, springbok and black wildebeest carcasses.
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All external fat was removed and discarded from the meat of each animal, vacuum packed per animal and frozen at -20°C until production day. Pork meat and pork back fat was sourced from
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Winelands Pork (Cape Town, South Africa) and kept frozen at -20°C until production day. The
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pH of the meat was measured upon thawing on the production days. 2.2 Starter culture and spices
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The starter culture Bitec LS 3 containing Lactobacillus curvatus and Staphylococcus carnosus
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was sourced from Freddy Hirsh (Cape Town, South Africa). A commercial spice pack, Salami Montana (Freddy Hirsh, Cape Town, South Africa) containing salt (46%), dextrose, irradiated
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spices, phosphates, starch (maize), vegetable protein (soya bean), sucrose, monosodium
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glutamate (flavor enhancer), ascorbic acid (1.23%), flavorings and spice extracts and curing agents (8% sodium nitrite, 90% salt, 2% sodium carbonate) packed separately was used
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according to producer’s specifications. 2.3 Salami preparation For each batch (5.211 kg), frozen meat (3.5 kg) and pork back fat (1.5 kg) was thawed for 24 hrs at ± 4°C and then mixed together with Salami Montana main pack (0.2 kg) and curing agents (11 g). The mixture was minced through a 5mm grinding plate before adding the starter culture solution (made by dissolving 1.25 g in 260 mL water for 30 min according to producer’s
ACCEPTED MANUSCRIPT specifications) and minced again. Before stuffing into casings, the raw batter (day 0) was measured for pH and sampled for proximate composition, salt content, aw and lipid oxidation. The raw batter was then stuffed into collagen oxygen and moisture permeable casings (60 mm diameter; ±12 cm length). All sausages were weighed before dipping into a spore solution of
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Bactoferm MOLD-600 containing Penicillium nalgiovense according to the producer’s
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specifications (25 g dissolved in 10 L of water). Eight batches of salami per species were
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produced independently (13 sausages per batch; each sausage weighing between 324 – 461 g before hanging). Each salami sausage was weighed before being randomly hung in a temperature
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and humidity controlled chamber (Stagionello Twin 100+100 kg, Food Transformation Systems,
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Crotone, Italy) for 23 days following the parameters given in Figure 1.
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One salami from each species’ batch was removed on different sampling days (4, 7, 12, 18 and 23), weighed to determine the weight loss, measured for pH and sampled for aw and lipid
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oxidation. Proximate composition and salt content were also measured at the end of production
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(day 23). Moisture content was calculated for each time point using the weight loss and experimental final moisture content. The experimental moisture content kinetics were fitted to
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the model of Henderson & Pabis (1961): 𝑀𝐶 (% 𝑑𝑚) = 𝑎 exp(−𝑘𝑡)
Where 𝑀𝐶 is the moisture content dry basis expressed in percentage of dry matter (dm), 𝑘 is the rate constant, 𝑎 is the model parameter and 𝑡 is the time. The model parameters were identified by nonlinear regression with a least square minimization procedure using the complement Excel ‘‘Solver”.
ACCEPTED MANUSCRIPT 2.4 Sample preparation One salami sausage from each species batch (n=8), was cut up and randomly allocated for different further analyses, vacuum packed and stored at -20°C (for aw, proximate composition and salt content samples) and -80°C (lipid oxidation). Before all analysis, raw batter and salami
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samples were homogenized (Golasecca (VA) Italy) for 30 secs and analysis done in duplicate.
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2.5 Analysis 2.5.1 pH
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The pH of the raw meat, raw batter before stuffing and salami was determined in triplicate using a Crison 200 pH meter fitted with an electrode probe. For salami, the electrode was inserted into
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the centre of the upper, middle and bottom sections of the salami.
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2.5.2 Proximate composition
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The samples were analyzed for moisture (Method 934.01) and ash (Method 942.05) content according to the AOAC (2002). The protein content was determined according to AOAC (1994)
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procedure 992.15, whereas fat content was determined using the chloroform/methanol (2:1) fat
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extraction method according to Lee, Trevino & Chaiyawat (1996). 2.5.3 Salt content
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A 0.3 g sample was weighed into a closed container and agitated under stirring for at least 2 hrs with 50 mL of 0.3 M nitric acid. The chloride concentration was determined using a 926 Chloride analyser (Sherwood Scientific, Cambridge, UK). 2.5.4 Water activity (aw) aw was measured at 25°C using an AquaLab 4TE water activity meter (Decagon, Washington, USA).
ACCEPTED MANUSCRIPT 2.5.5 Lipid oxidation Lipid oxidation was analysed by measuring the TBARS by acid-precipitation using the technique described by Descalzo et al. (2005). A 2 g sample was homogenized with 6.25 ml of trichloroacetic acid (2.8% w/v) and 6.25 ml of distilled water using a Polytron (Kinematica,
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Höganäs, Sweden) for 20 secs and filtered through a Whatman No. 1 filter paper. Of the filtrate,
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1 ml was transferred into 3 test tubes where 1 ml of 0.02 M thiobarbituric acid was added to 2
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test tubes and 1 ml of distilled water was added to the third test tube to function as the turbidity blank. Samples were vortexed and incubated at 70°C for 1 hr. After cooling, the absorbance was
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measured at 530 nm (Spectrostar Nano, BMG Labtech, Ortenberg, Germany). TBARS were
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calculated in mg equivalent malonaldehyde (MDA) per kg of meat using a calibration curve of
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1,1,3,3-tetramethoxypropan. 2.6 Statistical Analysis
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The experimental design was a 6x6 factorial in a completely randomized design with species
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(blesbok, eland, fallow deer, springbok, black wildebeest and pork) and processing time (0, 4, 7, 12, 18 and 23) as factors and batch (1-8) as a random factor. Data was analysed using MIXED
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PROC procedures of STATISTICA and t-Tests (PDIFF option) used to compare least square
3. Results
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means. Differences were significant at P≤0.05.
The levels of statistical significance (P-values) of the main effects and their interaction on the various chemical and physical parameters measured are indicated in Table 1. No interactions were noted for all measured attributes. Processing time was a significant factor on all measured attributes whilst species only affected (P≤0.05) pH of raw meat, moisture content and pH of salami.
ACCEPTED MANUSCRIPT 3.1 Drying kinetics The average changes in the weight loss during processing of salami produced from pork and different game species is shown in Figure 2 and the moisture content kinetics in Figure 3. There was no species effect on the weight loss and moisture content kinetics of salami. Weight loss
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reached 32.7% on average at the end of drying.
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3.2 pH of raw meat and salami
The pH of raw meat used to make salami and the progressive changes of salami pH thereafter
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and throughout processing is shown in Table 2. The pH of the raw meat did not differ among species except for black wildebeest which had a value of 6.30. Generally, pH dropped in all
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salami types and began increasing again after day 4 until the end of production. The lowest pH
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drop (on day 4) was observed in blesbok, fallow deer and springbok salami and was similar to that of pork salami. Black wildebeest salami consistently showed a higher pH than others
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throughout processing until day 12 when its pH became similar to the pH of eland salami. Then
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both showed higher pH than the others until day 18 when their pH became similar to the pH of pork salami. Final pH of the different types of salami ranged from 5.01 to 5.77 with pH of black
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wildebeest being significantly higher and fallow deer lower compared to pork.
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3.3 Proximate composition, salt content and aw The proximate composition of salami produced from pork and different game species is presented in Table 3. Proximate composition did not differ in raw batter as expected except that the moisture content of the raw batter from springbok was slightly lower than the other ones but similar to the raw batter from eland. The moisture content decreased in all types of salami with the highest being observed in black wildebeest and blesbok salami (38.5 and 37.8%, respectively) at the end of drying. Fat, protein content, ash and salt content increased in all
ACCEPTED MANUSCRIPT salami types from 22.6 to 35.7%; 16.4 to 23.6%; 3.09 to 4.60% and 2.23 to 3.23%, respectively on average. aw decreased throughout drying in all salami types from 0.970 to 0.908 on average (0.881-0.919) (Figure 2) and did not differ between species. 3.4 TBARS
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The average TBARS values of salami are shown in Figure 4. A general increase was noted
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throughout drying. No differences were noted between day 0 and day 4 as well as between day 18 and 23 of production. TBARS reached 0.85 mg MDA equivalent/kg in average.
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4. Discussion
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4.1 Composition
Moisture and salt content of the salami were in the range given by Zanardi, Ghidini, Conter, &
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Ianieri (2010) on commercial Milano type pork salami (n=27; 24.3-53.0% and 3.5-5.2% for
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moisture and salt content, respectively). Fermented products are known to contain fat contents
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varying from 25-45% in order to meet consumer expectations (Yilmaz et al., 2009). Zanardi et al. (2010) reported a range of 12 to 30% on commercial pork fermented sausages. The final fat
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content recorded for all salami types ranged from 32.5 to 37.3% (mean of 52.8-58.8% dm),
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which was higher than the results of Cenci-Goga et al. (2012) on swine and venison fermented sausages (n=5; 43.1-46.1% dm). On the other hand, it was lower than the results on wild boar salami (n=8; 45.9-51.1%) recorded by Paulsen et al. (2011). However, this is due to their lower moisture content (19.5% in average) and are not higher when expressed on a dry mass basis (57.0-63.4% dm). The fat content in this study is also in the range recorded on commercial venison and wild boar sausages (n=10; 19.4-52.6% dm) by Soriano, Cruz, Gómez, Mariscal, & García Ruiz (2006). Similar fat and moisture contents were found in gemsbok, kudu, springbok
ACCEPTED MANUSCRIPT and zebra salami (30.7% and 36.3% on average respectively) (Van Schalkwyk et al., 2011). Protein content at the end of drying (23.6% and 36.3% dm on average) is in the range (24.437.4%) reported in commercial pork fermented sausages by Zanardi et al. (2010). It was also in the range (32.4-57.5% dm) reported by Soriano et al. (2006) on commercial game fermented
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sausages and similar to the results (22.3% in average) on game salami of Van Schalkwyk et al.
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(2011) but lower than the results of Cenci-Goga et al. (2012) who found values ranging from
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44.9 to 49.8% dm.
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4.2 Impact of species on drying
During the production of fermented sausages, the progressive loss of weight is expected and
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depicts a decrease in moisture content. This moisture loss facilitates in reduction of the products’
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aw, which is an important safety hurdle against bacterial spoilage. In this study, the weight loss and moisture kinetics of pork salami did not differ from other salami types. However, despite
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similar drying kinetics, some difference in the final moisture content of salami was noted while it
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did not reflect in the aw or salt content or proximate composition. There is no data in literature comparing drying kinetics of salami from different game species or comparing them to pork.
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However, Van Schalkwyk et al. (2011) reported no differences in proximate composition and aw
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between gemsbok, kudu, springbok and zebra salami formulated the same way and dried under the same conditions which allows the hypothesis that they would have measured similar drying kinetics.
4.3 Impact of species on pH pH in fermented sausages decreases during fermentation due to the action of fermentative bacteria (Ordóñez et al., 1999). The extent and rate of pH decline depends on the amount of fermentable sugars used, starter culture types and fermenting temperatures. As noted in this
ACCEPTED MANUSCRIPT study, when drying progresses, pH begins to rise, indicating lactate and acetate utilization as well as ammonia production from amino acid break down (Ordóñez et al., 1999). The final pH attained by the salami in this study differed amongst species and ranged from 5.01 to 5.77. Other studies on fermented sausages from game have recorded pH values in the range of 4.4 to 6.8
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(Soriano et al., 2006; Todorov et al., 2007; García Ruiz, Mariscal, González Vinas, & Soriano,
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2010; Paulsen et al. 2011; Van Schalkwyk et al., 2011; Cenci-Goga et al., 2012; Utrilla et al.,
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2014; Maksimovic et al., 2018). Van Schalkwyk et al. (2011) noted species effect in the pH of game salami, with springbok salami showing higher pH (5.46) than gemsbok, kudu and zebra
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salami (4.98-5.00).
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The pH of all the salami types in this study fell below 5.3 with the exception of black wildebeest
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salami. This may be attributed to the initial high pH of black wildebeest meat. Game meat has a tendency to produce high pH meat during harvesting due to the sometimes stressful nature of
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harvesting (Hoffman & Wiklund, 2006). Black wildebeest is known to be easily stressed and in
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one study 58% of the harvested animals (n=12) where found to have a pH>6.00 (Shange, Makasi, Gouws, & Hoffman, 2018). Meat with high pH is considered difficult to ferment as it
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contains little lactate and sugar; impeding fermentation and providing a good medium for growth
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of spoilage bacteria (Lücke, 1994). The shelf life stability of black wildebeest salami (which showed aw and pH of 0.91 and 5.77, respectively) investigated in this study maybe questionable (Zukal & Incze, 2010). To ensure shelf life stability, black wildebeest may require longer drying periods to further reduce the aw. Alternatively, the addition of more fermentable sugars, longer fermentation periods or other starter cultures to ensure pH decline may be employed; the effects of these factors on high pH meat and the use thereof in salami production warrant further research.
ACCEPTED MANUSCRIPT 4.4 Impact of species on lipid oxidation Lipid oxidation is one of the major processes responsible for quality degradation in food products and leads to undesirable changes in flavor, colour and texture (Kanner, 2007). Products with TBARS values less than 1.0 mg MDA equivalent/kg are considered to be in an acceptable
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condition (Ospina-E, Rojano, Ochoa, Pérez-Álvarez, & Fernández-López, 2015) although this
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may depend on the method of analysis used and rancidity detection levels which may differ
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amongst products. Thus there may be need for sensory evaluation to determine rancidity detection levels. The TBARS values noted in this study are higher than those reported for
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pork/beef salchichon by Bruna, Ordóñez, Fernandez, Herranz, & de la Hoz (2001) and Zanardi et
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al. (2010) in Milano type pork sausages which ranged from 0.35 to 0.76 mg MDA equivalent/kg. However, similar TBARS values were reported by Utrilla et al. (2014) in fermented deer
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sausages made with pork meat (0.73 to 1.26 mg MDA equivalent/kg). It is worth mentioning that
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Bruna et al. (2001) and Utrilla et al. (2014) had different moisture content values, making the comparison difficult and thus differences in TBARS may not be attributed to species differences.
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There was no effect of game species and pork on TBARS. This may be attributed to the fact that
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pork back fat, which is the major source of lipids for oxidation in salami, was used for all salami types. However, the known high levels of heme iron in game meat which has pro-oxidant
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activities may have enhanced lipid oxidation in game salami, thus the need to investigate this aspect further.
5. Conclusion This study showed minimal species differences (P≤0.05) in the production of salami from different game meat species and the salami showed similar drying behavior and physicochemical attributes to the conventional pork based salami except in moisture content and pH.
ACCEPTED MANUSCRIPT Although moisture content differed slightly amongst species in both raw batter and end product, it was not reflected in the aw and thus may not have an effect on the shelf life of the salami. All salami types in this study were mainly characterized by a final aw of 0.91 in average and pH ranging from 5.01 to 5.28 with the exception of black wildebeest salami which showed a pH of
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5.77. When compared to pork, black wildebeest raw meat and salami consistently had a higher
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pH throughout drying, although it was similar to some game species at times. It is recommended
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that further work be done on salami from black wildebeest (meat with a high pH) to ensure shelf life stability. Future research should focus on evaluating the sensory characteristics of fermented
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game sausages as well as determining the flavor formation mechanisms (lipolysis, proteolysis) in
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order to identify and characterize any differences in salami from different game meat species and or specificity in comparison to conventional pork ones, as based on the findings in this study,
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applying the same processing techniques used for pork salami does not necessarily produce
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similar physico-chemical attributes when used on some game meat species.
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6. Acknowledgements
This research was funded by the South African Research Chairs Initiative (SARChI) and funded
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by the South African Department of Science and Technology, as administered by the National
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Research Foundation (NRF) of South Africa (UID: 84633). Any opinion, finding and conclusion or recommendation expressed in this material is that of the author(s) and the National Research Foundation does not accept any liability in this regard.
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ACCEPTED MANUSCRIPT Lee, C. M., Trevino, B., & Chaiyawat, M. (1996). A simple and rapid solvent extractionmethod for determining total lipids in fish tissue. Journal of AOAC International, 79, 487–492. Leistner, L., & Rodel, W. (1975). The significance of water activity for microorganisms in meats. In R. B. Duckworth (Ed.), Water relations of food (pp. 309–323). London: Academic
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Lin, Y., Huang, M., Zhou, G., Zou, Y., & Xu, X. (2011). Prooxidant Effects of the Combination of Green Tea Extract and Sodium Nitrite for Accelerating Lipolysis and Lipid Oxidation in
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Pepperoni during Storage. Journal of Food Science, 76(5).
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Ljung, P. E., Riley, S. J., & Ericsson, G. (2015). Game Meat Consumption Feeds Urban Support
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of Traditional Use of Natural Resources. Society and Natural Resources, 28(6), 657–669.
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Lorenzo, J. M., Montes, R., Purriños, L., & Franco, D. (2012). Effect of pork fat addition on the
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volatile compounds of foal dry-cured sausage. Meat Science, 91(4), 506–512.
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Lücke, F. (1994). Fermented meat products. Food Research International, 27, 299–307. Maksimovic, Z. A., Zunabovic-pichler, M., Kos, I., Mayrhofer, S., Hulak, N., Domig, K. J., &
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Mrkonjic, M. (2018). LWT - Food Science and Technology Microbiological hazards and potential of spontaneously fermented game meat sausages : A focus on lactic acid bacteria diversity. LWT - Food Science and Technology, 89 (November 2017), 418–426. McCrindle, C. M. E., Siegmund-Schultze, M., Heeb, A. W., Zárate, A. V., & Ramrajh, S. (2013). Improving food security and safety through use of edible by-products from wild game. Environment, Development and Sustainability, 15(5), 1245–1257.
ACCEPTED MANUSCRIPT Ordóñez, J. A., Hierro, E. M., Bruna, J. M., & de la Hoz, L. (1999). Changes in the Components of Dry-Fermented Sausages during Ripening. Critical Reviews in Food Science and Nutrition, 34(9), 329–367. Ospina-E, J. C., Rojano, B., Ochoa, O., Pérez-Álvarez, J. A., & Fernández-López, J. (2015).
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Development of frankfurter-type sausages with healthy lipid formulation and their
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nutritional, sensory and stability properties. European Journal of Lipid Science and Technology, 117(1), 122–131.
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Paulsen, P., Vali, S., & Bauer, F. (2011). Quality traits of wild boar mould-ripened salami
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manufactured with different selections of meat and fat tissue, and with and without bacterial
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starter cultures. Meat Science, 89(4), 486–490.
Ruiz, J. (2007). Ingredients. In F. Toldrá, Y. H. Hui, I. Astiasaran, W.-K. Nip, J. G. Sebranek,
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E.-T. Silveira, … R. Talon (Eds.), Handbook of fermented meat and Poultry (1st ed., pp.
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59–76). 2121 State Avenue, Ames, Iowa 50014, USA: Blackwell Publishing Ltd.
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Shange, N., Makasi, T. N., Gouws, P. A., & Ho, L. C. (2018). The influence of normal and high ultimate muscle pH on the microbiology and colour stability of previously frozen black
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wildebeest (Connochaetes gnou) meat, 135, 14–19. Soriano, A., Cruz, B., Gómez, L., Mariscal, C., & García Ruiz, A. (2006). Proteolysis, physicochemical characteristics and free fatty acid composition of dry sausages made with deer (Cervus elaphus) or wild boar (Sus scrofa) meat: A preliminary study. Food Chemistry, 96(2), 173–184. Todorov, S. D., Koep, K. S. C., Van Reenen, C., Hoffman, L. C., Slinde, E., & Dicks, L. M. T.
ACCEPTED MANUSCRIPT (2007). Production of salami from beef, horse, mutton, Blesbok (Damaliscus dorcas phillipsi) and Springbok (Antidorcas marsupialis) with bacteriocinogenic strains of Lactobacillus plantarum and Lactobacillus curvatus. Meat Science, 77(3), 405–12. Toldrá, F. (2002). Dry-cured meat products. (F. Toldrá, Ed.)Publications in Food Science and
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Nutrition (2nd ed.). Trumbull, Connecticut 06611 USA: Food and Nutrition Press, Inc.
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Utrilla, M. C., García Ruiz, A., & Soriano, A. (2014). Effect of partial reduction of pork meat on the physicochemical and sensory quality of dry ripened sausages: Development of a healthy
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venison salchichon. Meat Science, 98(4), 785–791.
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Van de Merwe, P. Van Der, Saayman, M., & Rossouw, R. (2015). The Economic Impact of
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Hunting in the Limpopo Province. Journal of Economic and Financial Sciences, 8, 223–
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242.
Van Schalkwyk, D. L., McMillin, K. W., Booyse, M., Witthuhn, R. C., & Hoffman, L. C.
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(2011). Physico-chemical, microbiological, textural and sensory attributes of matured game
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salami produced from springbok (Antidorcas marsupialis), gemsbok (Oryx gazella), kudu (Tragelaphus strepsiceros) and zebra (Equus burchelli) harvested in Namibia. Meat
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Science, 88(1), 36–44.
Van Schalkwyk, D. L., McMillin, K. W., Witthuhn, R. C., & Hoffman, L. C. (2010). The Contribution of Wildlife to Sustainable Natural Resource Utilization in Namibia: A Review. Sustainability, 2(11), 3479–3499. Yilmaz, I., Velioglu, H. M., Lachowicz, K., & Joanna, Ż. (2009). Fermented meat products. In I. Yilmaz (Ed.), Quality of meat and meat products (2nd ed., Vol. 661, pp. 1–16). Kerala:
ACCEPTED MANUSCRIPT Transworld Research Network. Zanardi, E., Ghidini, S., Conter, M., & Ianieri, A. (2010). Mineral composition of Italian salami and effect of NaCl partial replacement on compositional , physico-chemical and sensory
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parameters. Meat Science, 86(3), 742–747.
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Zukal, E., & Incze, K. (2010). Drying. In F. Toldrá (Ed.), Handbook of meat processing (pp.
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219–229). Singapore: Markomo Print Media Pte Ltd.
ACCEPTED MANUSCRIPT Table 1: Level of statistical significance (P-values) for the main effects of species, processing time and their interactions Species
Processing time
Raw meat pH Weight loss Moisture Fat Protein Ash Salt content Water activity pH TBARS
<.0001 0.2364 0.0100 0.1422 0.6950 0.5144 0.4359 0.5758 <.0001 0.0663
<.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001
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P-values in bold indicate a significant difference at P≤0.05
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Species x Processing time 0.3505 0.4500 0.1732 0.8054 0.6351 0.6079 0.0962 0.3603 0.7040
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Attributes
ACCEPTED MANUSCRIPT Table 2: Means and standard errors (n=8) for pH of game and pork raw meat and salami during processing Pork
Blesbok
Eland
Fallow
0.07 Day 4
4.93bc 0.06
Day 7
5.03b 0.07
Day 12
5.01b
Day 23
0.06 ± 4.78bc 0.05 ± 4.86b 0.04
5.20ab
± 4.97b
0.07 5.19b
0.04
± 5.02b 0.07
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0.07 abc
± 4.70c
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Day 18
0.08
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0.09
± 5.75b
0.09 ± 5.86b
0.04 ± 5.78b
0.06 ± 5.05b 0.09 ± 5.07b 0.07
± 5.20a 0.11
± 5.29a 0.07 ± 5.28ab 0.06
wildebeest ± 5.63b
± 5.78b
0.08
0.07
± 4.87c
± 4.76c
0.04
± 4.88c 0.11
± 4.97b 0.10 ± 5.09b 0.09 ± 5.01c 0.18
± 6.30a ± 0.10
0.05
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5.88b
± 5.66b
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Day 0
0.05
± 5.73b
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0.09
± 5.64b
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5.80b
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Raw meat
deer
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time
Springbok Black
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Sampling
± 6.24a ± 0.08
± 5.36a ± 0.20
0.08 ± 4.85bc
± 5.31a ± 0.22
0.07 ± 4.86b
± 5.44a ± 0.25
0.07 ± 4.96b
± 5.51a ± 0.23
0.06 ± 5.04bc
± 5.77a ± 0.26
0.09
Means with different superscripts across rows are significantly different (P≤0.05) between
species at each sampling time
ACCEPTED MANUSCRIPT Table 3: Means and standard errors (n=8) for proximate and salt content of game and pork salami during processing
58.8a±0.8 0
58.4a±0.4 0
57.9a±0.4 2
56.2ab±1.3 2
23
34.3b±0.9 6
37.8a± 0.96
35.1b±0.5 9
0
23.2± 0.51
20.7± 1.80
20.7± 1.29
23
36.1± 0.48
36.4± 2.71
0
16.2± 0.93
16.2± 1.29
23
25.8±2.30 21.2± 2.13
0
Salt content (%)
3.04± 0.07
3.02± 0.04
Springbo k
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0
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Ash (%)
Fallow deer
57.9a± 0.69
33.2b±0.9 4
38.5a± 0.66
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55.8b±0.6 3
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34.4b±0.3 3
Black wildebees t
23.5± 3.18 23.2± 1.61
24.0± 2.49
37.2± 2.96 37.3± 3.68
32.5± 3.07
17.0±0.97 16.1± 1.74 16.6± 1.50
16.0± 1.83
25.0± 1.96
24.4± 2.90
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Protein (%)
Eland
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Fat (%)
Blesbok
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Moisture (%)
Pork
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Attribute Samplin s g day
34.4± 2.56
22.2± 2.65 23.1±3.11
3.03±0.04 3.14± 0.11 3.04± 0.05
3.29± 0.29 4.57± 0.03
23
4.94±0.12 4.51± 0.06
4.46± 0.07
0
2.22± 0.11
2.19±0.06 2.24± 0.10 2.24± 0.06
2.11± 0.13
23
3.29±0.03 3.24± 0.05
3.10± 0.06
3.33± 0.08
2.38± 0.08
4.54± 0.04 4.57±0.10
3.27± 0.04 3.14±0.11
ACCEPTED MANUSCRIPT ab
Means with different superscripts across rows are significantly different (P≤0.05) between
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species at each sampling day
ACCEPTED MANUSCRIPT Highlights
T IP CR US AN M ED PT
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Different game meat species were used to produce independent batches of salami and compared to a conventional pork based salami. Minimal species differences were noted as well as similar drying kinetics and physico-chemical attributes to the conventional pork salami. Black wildebeest meat had a higher initial pH which translated to differences in some physicochemical attributes of the salami with conventional pork salami.
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Figure 1
Figure 2
Figure 3
Figure 4
Chido Chakanya, Elodie Arnaud, Voster Muchenje, Louwrens C. Hoffman PII: DOI: Reference:
S0309-1740(18)30409-1 doi:10.1016/j.meatsci.2018.07.034 MESC 7648
To appear in:
Meat Science
Received date: Revised date: Accepted date:
16 April 2018 22 June 2018 30 July 2018
Please cite this article as: Chido Chakanya, Elodie Arnaud, Voster Muchenje, Louwrens C. Hoffman , Changes in the physico-chemical attributes through processing of salami made from blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) in comparison to pork. Mesc (2018), doi:10.1016/j.meatsci.2018.07.034
This is a PDF file of an unedited manuscript that has been accepted for publication. As a service to our customers we are providing this early version of the manuscript. The manuscript will undergo copyediting, typesetting, and review of the resulting proof before it is published in its final form. Please note that during the production process errors may be discovered which could affect the content, and all legal disclaimers that apply to the journal pertain.
ACCEPTED MANUSCRIPT Changes in the physico-chemical attributes through processing of salami made from blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) in
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comparison to pork
1
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Chido Chakanya1, Elodie Arnaud2,3,4, Voster Muchenje1 and Louwrens C. Hoffman2,* Department of Livestock and Pasture Science, University of Fort Hare, Private Bag X 1314,
2
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Alice 5700, South Africa
Department of Animal Sciences, Stellenbosch University, Private Bag X1 Matieland,
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Stellenbosch 7602, South Africa
CIRAD, UMR QualiSud, 7602 Stellenbosch, South Africa
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Qualisud, Univ Montpellier, CIRAD, Montpellier SupAgro, Université d'Avignon, Université
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3
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de La Réunion, Montpellier, France
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*Corresponding author: [email protected]
ACCEPTED MANUSCRIPT Abstract Drying kinetics and changes in proximate composition, pH, salt content, water activity (aw) and lipid oxidation through processing of salami made using five different game meat species were evaluated and compared to pork. Eight batches of salami from each species were made and
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sampled for analysis throughout processing. Processing time was a significant factor on all
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measured attributes whilst species affected (P≤0.05) pH and moisture but not drying kinetics.
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Black wildebeest meat exhibited higher (P≤0.05) pH than pork and other game meat (6.30 vs 5.63-5.80), which translated to higher (P≤0.05) salami pH throughout and at the end of
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processing (5.77). Final pH of all other salami ranged from 5.01 to 5.28, aw ranged from 0.88 to
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0.92. TBARS remained lower than 1 mg MDA equivalent/kg. The study suggests that salami from these game species, excluding black wildebeest, can be produced using the same processing
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parameters as conventional pork salami and obtaining similar physico-chemical attributes.
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Key words: fermented sausages; game meat; lipid oxidation; TBARS; drying kinetics
ACCEPTED MANUSCRIPT 1. Introduction The favorable nutritional and physico-chemical profile of meat from game species has been shown over the years (Hoffman & Cawthorn, 2013). Furthermore, an increase in the production of meat from game species has been noted in some regions, giving opportunity for the expansion
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of the game meat industry (Van Schalkwyk, McMillin, Witthuhn, & Hoffman, 2010; Ljung,
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Riley, & Ericsson, 2015). In South Africa, most game meat is a by-product of trophy hunting/and or culling activities (McCrindle, Siegmund-Schultze, Heeb, Zárate, & Ramrajh, 2013). Game
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meat is commonly used for the production of biltong made from salted and dried meat strips (Jones, Arnaud, Gouws, & Hoffman, 2017) and fermented sausage (salami) (Van Schalkwyk et
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al., 2011; Van de Merwe, Saayman, & Rossouw, 2015) as it is sometimes perceived as being
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tough and hard to cook (Hoffman, Muller, Schutte, & Crafford, 2004).
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Fermented sausages are popularly produced using pork meat and fat (Ruiz, 2007; Yilmaz, Velioglu, Lachowicz, & Joanna, 2009). Their production can be separated into three distinct
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phases; formulation, fermentation and drying/ripening. Formulation entails the preparation of
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seasoned, minced raw meat and fat for encasing whilst fermentation is crucial for the production of lactic acid which lowers pH and drying/ripening ensures the lowering of water activity (aw)
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through moisture loss (Ordóñez, Hierro, Bruna, & de la Hoz, 1999). The shelf stability of fermented sausages is based on the combined hurdle techniques of lowering pH and aw in order to create an environment that impedes growth of spoilage bacteria (Toldrá, 2002). Generally, the relationship between aw and pH of fermented products should be inversely proportional to ensure shelf life stability, i.e the higher the final pH, the lower the aw should be (Zukal & Incze, 2010). Meat products are considered shelf stable when they have a pH lower than 5 or aw lower than 0.91 or a combination of pH lower than 5.2 and aw below 0.95 (Leistner & Rodel, 1975).
ACCEPTED MANUSCRIPT However, there is still some debate about the precise pH and aw as noted by Abunyewa, Laing, Hugo, & Viljoen (2000). It is known that the characteristics of the raw material used impacts on the quality of the end product in sausage fermentation (Ruiz, 2007). Game meat is known to have lower intra muscular
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fat, higher amounts of polyunsaturated fatty acids as well as higher heme iron and myoglobin
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contents (Hoffman & Wiklund, 2006) compared to pork. These characteristics may have a significant bearing on its fermentation quality. Production of fermented sausages using game
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meat is relatively unexplored especially on wildlife species found in Southern Africa. Studies in which fermented sausages were produced using game meat and pork as a fat source have focused
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on wild boar and deer (Paulsen, Vali, & Bauer, 2011; Cenci-Goga et al., 2012; Utrilla, García
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Ruiz, & Soriano, 2014; Maskimovic et al., 2018). Todorov et al. (2007) and Van Schalkwyk et al. (2011) looked at the physico-chemical, microbial and sensory quality of fermented sausages
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made using springbok (Antidorcas marsupialis), gemsbok (Oryx gazella), kudu (Tragelaphus
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strepsiceros), zebra (Equus burchelli) and blesbok (Damaliscus pygargus phillipsi). However, little is known on the physico-chemical changes that occur during their processing. These
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changes are critical in developing guidelines for the production of fermented sausages for
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individual species with consistently high quality. Therefore this study aimed at characterizing the changes that occur during production of salami made using species commonly harvested in South Africa ie. blesbok (Damaliscus pygargus phillipsi), eland (Taurotragus oryx), fallow deer (Dama dama), springbok (Antidorcas marsupialis) and black wildebeest (Connochaetes gnou) and comparing them to pork salami.
ACCEPTED MANUSCRIPT 2. Materials and methods 2.1 Raw meat Meat was sourced from the skeletal muscles and trimmings after removal of major muscles (semitendinosus, biceps femoris, infra and supra spinatus, semimembranosus, longissimus
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thoracis et lamborum) of blesbok, eland, fallow deer, springbok and black wildebeest carcasses.
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All external fat was removed and discarded from the meat of each animal, vacuum packed per animal and frozen at -20°C until production day. Pork meat and pork back fat was sourced from
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Winelands Pork (Cape Town, South Africa) and kept frozen at -20°C until production day. The
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pH of the meat was measured upon thawing on the production days. 2.2 Starter culture and spices
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The starter culture Bitec LS 3 containing Lactobacillus curvatus and Staphylococcus carnosus
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was sourced from Freddy Hirsh (Cape Town, South Africa). A commercial spice pack, Salami Montana (Freddy Hirsh, Cape Town, South Africa) containing salt (46%), dextrose, irradiated
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spices, phosphates, starch (maize), vegetable protein (soya bean), sucrose, monosodium
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glutamate (flavor enhancer), ascorbic acid (1.23%), flavorings and spice extracts and curing agents (8% sodium nitrite, 90% salt, 2% sodium carbonate) packed separately was used
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according to producer’s specifications. 2.3 Salami preparation For each batch (5.211 kg), frozen meat (3.5 kg) and pork back fat (1.5 kg) was thawed for 24 hrs at ± 4°C and then mixed together with Salami Montana main pack (0.2 kg) and curing agents (11 g). The mixture was minced through a 5mm grinding plate before adding the starter culture solution (made by dissolving 1.25 g in 260 mL water for 30 min according to producer’s
ACCEPTED MANUSCRIPT specifications) and minced again. Before stuffing into casings, the raw batter (day 0) was measured for pH and sampled for proximate composition, salt content, aw and lipid oxidation. The raw batter was then stuffed into collagen oxygen and moisture permeable casings (60 mm diameter; ±12 cm length). All sausages were weighed before dipping into a spore solution of
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Bactoferm MOLD-600 containing Penicillium nalgiovense according to the producer’s
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specifications (25 g dissolved in 10 L of water). Eight batches of salami per species were
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produced independently (13 sausages per batch; each sausage weighing between 324 – 461 g before hanging). Each salami sausage was weighed before being randomly hung in a temperature
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and humidity controlled chamber (Stagionello Twin 100+100 kg, Food Transformation Systems,
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Crotone, Italy) for 23 days following the parameters given in Figure 1.
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One salami from each species’ batch was removed on different sampling days (4, 7, 12, 18 and 23), weighed to determine the weight loss, measured for pH and sampled for aw and lipid
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oxidation. Proximate composition and salt content were also measured at the end of production
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(day 23). Moisture content was calculated for each time point using the weight loss and experimental final moisture content. The experimental moisture content kinetics were fitted to
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the model of Henderson & Pabis (1961): 𝑀𝐶 (% 𝑑𝑚) = 𝑎 exp(−𝑘𝑡)
Where 𝑀𝐶 is the moisture content dry basis expressed in percentage of dry matter (dm), 𝑘 is the rate constant, 𝑎 is the model parameter and 𝑡 is the time. The model parameters were identified by nonlinear regression with a least square minimization procedure using the complement Excel ‘‘Solver”.
ACCEPTED MANUSCRIPT 2.4 Sample preparation One salami sausage from each species batch (n=8), was cut up and randomly allocated for different further analyses, vacuum packed and stored at -20°C (for aw, proximate composition and salt content samples) and -80°C (lipid oxidation). Before all analysis, raw batter and salami
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samples were homogenized (Golasecca (VA) Italy) for 30 secs and analysis done in duplicate.
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2.5 Analysis 2.5.1 pH
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The pH of the raw meat, raw batter before stuffing and salami was determined in triplicate using a Crison 200 pH meter fitted with an electrode probe. For salami, the electrode was inserted into
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the centre of the upper, middle and bottom sections of the salami.
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2.5.2 Proximate composition
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The samples were analyzed for moisture (Method 934.01) and ash (Method 942.05) content according to the AOAC (2002). The protein content was determined according to AOAC (1994)
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procedure 992.15, whereas fat content was determined using the chloroform/methanol (2:1) fat
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extraction method according to Lee, Trevino & Chaiyawat (1996). 2.5.3 Salt content
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A 0.3 g sample was weighed into a closed container and agitated under stirring for at least 2 hrs with 50 mL of 0.3 M nitric acid. The chloride concentration was determined using a 926 Chloride analyser (Sherwood Scientific, Cambridge, UK). 2.5.4 Water activity (aw) aw was measured at 25°C using an AquaLab 4TE water activity meter (Decagon, Washington, USA).
ACCEPTED MANUSCRIPT 2.5.5 Lipid oxidation Lipid oxidation was analysed by measuring the TBARS by acid-precipitation using the technique described by Descalzo et al. (2005). A 2 g sample was homogenized with 6.25 ml of trichloroacetic acid (2.8% w/v) and 6.25 ml of distilled water using a Polytron (Kinematica,
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Höganäs, Sweden) for 20 secs and filtered through a Whatman No. 1 filter paper. Of the filtrate,
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1 ml was transferred into 3 test tubes where 1 ml of 0.02 M thiobarbituric acid was added to 2
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test tubes and 1 ml of distilled water was added to the third test tube to function as the turbidity blank. Samples were vortexed and incubated at 70°C for 1 hr. After cooling, the absorbance was
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measured at 530 nm (Spectrostar Nano, BMG Labtech, Ortenberg, Germany). TBARS were
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calculated in mg equivalent malonaldehyde (MDA) per kg of meat using a calibration curve of
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1,1,3,3-tetramethoxypropan. 2.6 Statistical Analysis
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The experimental design was a 6x6 factorial in a completely randomized design with species
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(blesbok, eland, fallow deer, springbok, black wildebeest and pork) and processing time (0, 4, 7, 12, 18 and 23) as factors and batch (1-8) as a random factor. Data was analysed using MIXED
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PROC procedures of STATISTICA and t-Tests (PDIFF option) used to compare least square
3. Results
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means. Differences were significant at P≤0.05.
The levels of statistical significance (P-values) of the main effects and their interaction on the various chemical and physical parameters measured are indicated in Table 1. No interactions were noted for all measured attributes. Processing time was a significant factor on all measured attributes whilst species only affected (P≤0.05) pH of raw meat, moisture content and pH of salami.
ACCEPTED MANUSCRIPT 3.1 Drying kinetics The average changes in the weight loss during processing of salami produced from pork and different game species is shown in Figure 2 and the moisture content kinetics in Figure 3. There was no species effect on the weight loss and moisture content kinetics of salami. Weight loss
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reached 32.7% on average at the end of drying.
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3.2 pH of raw meat and salami
The pH of raw meat used to make salami and the progressive changes of salami pH thereafter
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and throughout processing is shown in Table 2. The pH of the raw meat did not differ among species except for black wildebeest which had a value of 6.30. Generally, pH dropped in all
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salami types and began increasing again after day 4 until the end of production. The lowest pH
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drop (on day 4) was observed in blesbok, fallow deer and springbok salami and was similar to that of pork salami. Black wildebeest salami consistently showed a higher pH than others
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throughout processing until day 12 when its pH became similar to the pH of eland salami. Then
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both showed higher pH than the others until day 18 when their pH became similar to the pH of pork salami. Final pH of the different types of salami ranged from 5.01 to 5.77 with pH of black
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wildebeest being significantly higher and fallow deer lower compared to pork.
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3.3 Proximate composition, salt content and aw The proximate composition of salami produced from pork and different game species is presented in Table 3. Proximate composition did not differ in raw batter as expected except that the moisture content of the raw batter from springbok was slightly lower than the other ones but similar to the raw batter from eland. The moisture content decreased in all types of salami with the highest being observed in black wildebeest and blesbok salami (38.5 and 37.8%, respectively) at the end of drying. Fat, protein content, ash and salt content increased in all
ACCEPTED MANUSCRIPT salami types from 22.6 to 35.7%; 16.4 to 23.6%; 3.09 to 4.60% and 2.23 to 3.23%, respectively on average. aw decreased throughout drying in all salami types from 0.970 to 0.908 on average (0.881-0.919) (Figure 2) and did not differ between species. 3.4 TBARS
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The average TBARS values of salami are shown in Figure 4. A general increase was noted
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throughout drying. No differences were noted between day 0 and day 4 as well as between day 18 and 23 of production. TBARS reached 0.85 mg MDA equivalent/kg in average.
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4. Discussion
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4.1 Composition
Moisture and salt content of the salami were in the range given by Zanardi, Ghidini, Conter, &
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Ianieri (2010) on commercial Milano type pork salami (n=27; 24.3-53.0% and 3.5-5.2% for
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moisture and salt content, respectively). Fermented products are known to contain fat contents
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varying from 25-45% in order to meet consumer expectations (Yilmaz et al., 2009). Zanardi et al. (2010) reported a range of 12 to 30% on commercial pork fermented sausages. The final fat
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content recorded for all salami types ranged from 32.5 to 37.3% (mean of 52.8-58.8% dm),
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which was higher than the results of Cenci-Goga et al. (2012) on swine and venison fermented sausages (n=5; 43.1-46.1% dm). On the other hand, it was lower than the results on wild boar salami (n=8; 45.9-51.1%) recorded by Paulsen et al. (2011). However, this is due to their lower moisture content (19.5% in average) and are not higher when expressed on a dry mass basis (57.0-63.4% dm). The fat content in this study is also in the range recorded on commercial venison and wild boar sausages (n=10; 19.4-52.6% dm) by Soriano, Cruz, Gómez, Mariscal, & García Ruiz (2006). Similar fat and moisture contents were found in gemsbok, kudu, springbok
ACCEPTED MANUSCRIPT and zebra salami (30.7% and 36.3% on average respectively) (Van Schalkwyk et al., 2011). Protein content at the end of drying (23.6% and 36.3% dm on average) is in the range (24.437.4%) reported in commercial pork fermented sausages by Zanardi et al. (2010). It was also in the range (32.4-57.5% dm) reported by Soriano et al. (2006) on commercial game fermented
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sausages and similar to the results (22.3% in average) on game salami of Van Schalkwyk et al.
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(2011) but lower than the results of Cenci-Goga et al. (2012) who found values ranging from
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44.9 to 49.8% dm.
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4.2 Impact of species on drying
During the production of fermented sausages, the progressive loss of weight is expected and
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depicts a decrease in moisture content. This moisture loss facilitates in reduction of the products’
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aw, which is an important safety hurdle against bacterial spoilage. In this study, the weight loss and moisture kinetics of pork salami did not differ from other salami types. However, despite
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similar drying kinetics, some difference in the final moisture content of salami was noted while it
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did not reflect in the aw or salt content or proximate composition. There is no data in literature comparing drying kinetics of salami from different game species or comparing them to pork.
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However, Van Schalkwyk et al. (2011) reported no differences in proximate composition and aw
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between gemsbok, kudu, springbok and zebra salami formulated the same way and dried under the same conditions which allows the hypothesis that they would have measured similar drying kinetics.
4.3 Impact of species on pH pH in fermented sausages decreases during fermentation due to the action of fermentative bacteria (Ordóñez et al., 1999). The extent and rate of pH decline depends on the amount of fermentable sugars used, starter culture types and fermenting temperatures. As noted in this
ACCEPTED MANUSCRIPT study, when drying progresses, pH begins to rise, indicating lactate and acetate utilization as well as ammonia production from amino acid break down (Ordóñez et al., 1999). The final pH attained by the salami in this study differed amongst species and ranged from 5.01 to 5.77. Other studies on fermented sausages from game have recorded pH values in the range of 4.4 to 6.8
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(Soriano et al., 2006; Todorov et al., 2007; García Ruiz, Mariscal, González Vinas, & Soriano,
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2010; Paulsen et al. 2011; Van Schalkwyk et al., 2011; Cenci-Goga et al., 2012; Utrilla et al.,
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2014; Maksimovic et al., 2018). Van Schalkwyk et al. (2011) noted species effect in the pH of game salami, with springbok salami showing higher pH (5.46) than gemsbok, kudu and zebra
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salami (4.98-5.00).
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The pH of all the salami types in this study fell below 5.3 with the exception of black wildebeest
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salami. This may be attributed to the initial high pH of black wildebeest meat. Game meat has a tendency to produce high pH meat during harvesting due to the sometimes stressful nature of
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harvesting (Hoffman & Wiklund, 2006). Black wildebeest is known to be easily stressed and in
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one study 58% of the harvested animals (n=12) where found to have a pH>6.00 (Shange, Makasi, Gouws, & Hoffman, 2018). Meat with high pH is considered difficult to ferment as it
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contains little lactate and sugar; impeding fermentation and providing a good medium for growth
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of spoilage bacteria (Lücke, 1994). The shelf life stability of black wildebeest salami (which showed aw and pH of 0.91 and 5.77, respectively) investigated in this study maybe questionable (Zukal & Incze, 2010). To ensure shelf life stability, black wildebeest may require longer drying periods to further reduce the aw. Alternatively, the addition of more fermentable sugars, longer fermentation periods or other starter cultures to ensure pH decline may be employed; the effects of these factors on high pH meat and the use thereof in salami production warrant further research.
ACCEPTED MANUSCRIPT 4.4 Impact of species on lipid oxidation Lipid oxidation is one of the major processes responsible for quality degradation in food products and leads to undesirable changes in flavor, colour and texture (Kanner, 2007). Products with TBARS values less than 1.0 mg MDA equivalent/kg are considered to be in an acceptable
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condition (Ospina-E, Rojano, Ochoa, Pérez-Álvarez, & Fernández-López, 2015) although this
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may depend on the method of analysis used and rancidity detection levels which may differ
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amongst products. Thus there may be need for sensory evaluation to determine rancidity detection levels. The TBARS values noted in this study are higher than those reported for
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pork/beef salchichon by Bruna, Ordóñez, Fernandez, Herranz, & de la Hoz (2001) and Zanardi et
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al. (2010) in Milano type pork sausages which ranged from 0.35 to 0.76 mg MDA equivalent/kg. However, similar TBARS values were reported by Utrilla et al. (2014) in fermented deer
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sausages made with pork meat (0.73 to 1.26 mg MDA equivalent/kg). It is worth mentioning that
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Bruna et al. (2001) and Utrilla et al. (2014) had different moisture content values, making the comparison difficult and thus differences in TBARS may not be attributed to species differences.
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There was no effect of game species and pork on TBARS. This may be attributed to the fact that
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pork back fat, which is the major source of lipids for oxidation in salami, was used for all salami types. However, the known high levels of heme iron in game meat which has pro-oxidant
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activities may have enhanced lipid oxidation in game salami, thus the need to investigate this aspect further.
5. Conclusion This study showed minimal species differences (P≤0.05) in the production of salami from different game meat species and the salami showed similar drying behavior and physicochemical attributes to the conventional pork based salami except in moisture content and pH.
ACCEPTED MANUSCRIPT Although moisture content differed slightly amongst species in both raw batter and end product, it was not reflected in the aw and thus may not have an effect on the shelf life of the salami. All salami types in this study were mainly characterized by a final aw of 0.91 in average and pH ranging from 5.01 to 5.28 with the exception of black wildebeest salami which showed a pH of
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5.77. When compared to pork, black wildebeest raw meat and salami consistently had a higher
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pH throughout drying, although it was similar to some game species at times. It is recommended
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that further work be done on salami from black wildebeest (meat with a high pH) to ensure shelf life stability. Future research should focus on evaluating the sensory characteristics of fermented
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game sausages as well as determining the flavor formation mechanisms (lipolysis, proteolysis) in
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order to identify and characterize any differences in salami from different game meat species and or specificity in comparison to conventional pork ones, as based on the findings in this study,
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applying the same processing techniques used for pork salami does not necessarily produce
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similar physico-chemical attributes when used on some game meat species.
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6. Acknowledgements
This research was funded by the South African Research Chairs Initiative (SARChI) and funded
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by the South African Department of Science and Technology, as administered by the National
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Research Foundation (NRF) of South Africa (UID: 84633). Any opinion, finding and conclusion or recommendation expressed in this material is that of the author(s) and the National Research Foundation does not accept any liability in this regard.
ACCEPTED MANUSCRIPT 7. References Abunyewa, A. A. O., Laing, E., Hugo, A., & Viljoen, B. (2000). The population change of yeasts in commercial salami. Food Microbiology, 17(4), 429–438.
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ACCEPTED MANUSCRIPT Table 1: Level of statistical significance (P-values) for the main effects of species, processing time and their interactions Species
Processing time
Raw meat pH Weight loss Moisture Fat Protein Ash Salt content Water activity pH TBARS
<.0001 0.2364 0.0100 0.1422 0.6950 0.5144 0.4359 0.5758 <.0001 0.0663
<.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001 <.0001
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IP CR
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PT
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M
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P-values in bold indicate a significant difference at P≤0.05
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Species x Processing time 0.3505 0.4500 0.1732 0.8054 0.6351 0.6079 0.0962 0.3603 0.7040
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Attributes
ACCEPTED MANUSCRIPT Table 2: Means and standard errors (n=8) for pH of game and pork raw meat and salami during processing Pork
Blesbok
Eland
Fallow
0.07 Day 4
4.93bc 0.06
Day 7
5.03b 0.07
Day 12
5.01b
Day 23
0.06 ± 4.78bc 0.05 ± 4.86b 0.04
5.20ab
± 4.97b
0.07 5.19b
0.04
± 5.02b 0.07
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0.07 abc
± 4.70c
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Day 18
0.08
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0.09
± 5.75b
0.09 ± 5.86b
0.04 ± 5.78b
0.06 ± 5.05b 0.09 ± 5.07b 0.07
± 5.20a 0.11
± 5.29a 0.07 ± 5.28ab 0.06
wildebeest ± 5.63b
± 5.78b
0.08
0.07
± 4.87c
± 4.76c
0.04
± 4.88c 0.11
± 4.97b 0.10 ± 5.09b 0.09 ± 5.01c 0.18
± 6.30a ± 0.10
0.05
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5.88b
± 5.66b
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Day 0
0.05
± 5.73b
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0.09
± 5.64b
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5.80b
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Raw meat
deer
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time
Springbok Black
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Sampling
± 6.24a ± 0.08
± 5.36a ± 0.20
0.08 ± 4.85bc
± 5.31a ± 0.22
0.07 ± 4.86b
± 5.44a ± 0.25
0.07 ± 4.96b
± 5.51a ± 0.23
0.06 ± 5.04bc
± 5.77a ± 0.26
0.09
Means with different superscripts across rows are significantly different (P≤0.05) between
species at each sampling time
ACCEPTED MANUSCRIPT Table 3: Means and standard errors (n=8) for proximate and salt content of game and pork salami during processing
58.8a±0.8 0
58.4a±0.4 0
57.9a±0.4 2
56.2ab±1.3 2
23
34.3b±0.9 6
37.8a± 0.96
35.1b±0.5 9
0
23.2± 0.51
20.7± 1.80
20.7± 1.29
23
36.1± 0.48
36.4± 2.71
0
16.2± 0.93
16.2± 1.29
23
25.8±2.30 21.2± 2.13
0
Salt content (%)
3.04± 0.07
3.02± 0.04
Springbo k
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0
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Ash (%)
Fallow deer
57.9a± 0.69
33.2b±0.9 4
38.5a± 0.66
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55.8b±0.6 3
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34.4b±0.3 3
Black wildebees t
23.5± 3.18 23.2± 1.61
24.0± 2.49
37.2± 2.96 37.3± 3.68
32.5± 3.07
17.0±0.97 16.1± 1.74 16.6± 1.50
16.0± 1.83
25.0± 1.96
24.4± 2.90
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M
Protein (%)
Eland
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Fat (%)
Blesbok
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Moisture (%)
Pork
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Attribute Samplin s g day
34.4± 2.56
22.2± 2.65 23.1±3.11
3.03±0.04 3.14± 0.11 3.04± 0.05
3.29± 0.29 4.57± 0.03
23
4.94±0.12 4.51± 0.06
4.46± 0.07
0
2.22± 0.11
2.19±0.06 2.24± 0.10 2.24± 0.06
2.11± 0.13
23
3.29±0.03 3.24± 0.05
3.10± 0.06
3.33± 0.08
2.38± 0.08
4.54± 0.04 4.57±0.10
3.27± 0.04 3.14±0.11
ACCEPTED MANUSCRIPT ab
Means with different superscripts across rows are significantly different (P≤0.05) between
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PT
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M
AN
US
CR
IP
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species at each sampling day
ACCEPTED MANUSCRIPT Highlights
T IP CR US AN M ED PT
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Different game meat species were used to produce independent batches of salami and compared to a conventional pork based salami. Minimal species differences were noted as well as similar drying kinetics and physico-chemical attributes to the conventional pork salami. Black wildebeest meat had a higher initial pH which translated to differences in some physicochemical attributes of the salami with conventional pork salami.
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Figure 1
Figure 2
Figure 3
Figure 4