Nuk Przyrod Technologie ISSN 1897-7820 http://www.npt.up-poznn.net Dził: Nuki o Żywności i Żywieniu Copyright Wydwnictwo Uniwersytetu Przyrodniczego w Poznniu 2009 Tom 3 Zeszyt 4 HANNA GAJEWSKA-SZCZERBAL 1, MIROSŁAWA KRZYWDZIŃSKA-BARTKOWIAK 1, DONATA JARMOŁOWSKA-JURCZYSZYN 2 1 Institute of Met Technology Poznń University of Life Sciences 2 Chir nd Section of Clinicl Pthomorphology Poznń University of Medicl Sciences CHANGES IN PORK MUSCLES STRUCTURE PARAMETERS IN THE COURSE OF CURING AND PASTEURISATION Summry. The im of the study ws to perform comprtive evlution of the histologicl structure of the following pork hm muscles: the semimembrnosus muscle (musculus semimembrnosus) nd the qudriceps muscle of thigh (musculus qudriceps femoris) nd to compre the extent of these chnges following the injection with curing brine under the pressure of 0.5 MP, plsticiztion (mssging) in vcuum conditions nd psteuristion. Muscles for investigtions were collected from crcsses 24 h fter slughter. The injected curing brine contined, mong others, sodium chloride nd polyphosphtes. In order to perform histometric mesurements t ech phse of the experiment, histologicl preprtions were mde which were stined with the ssistnce of the HE method (hemtoxylin nd eosin). Mesurements were conducted with the ssistnce of the computer imge nlysis (CIA) using for this purpose MultiScn softwre. Microscope pictures were chrcterised on the bsis of the following muscle fibres prmeters: cell cross-section re, cell circumference, s well s Feret s H nd V dimeters. In ddition, percentge proportion of muscle fibres nd their quntity in the nlysed field of vision were clculted. The shpe of the fibres cross-section ws determined on the bsis of the rtio of Feret s H nd V dimeters. The results of ll mesurements were tested for the significnce level t p = 0.05. The performed comprtive nlysis of structure prmeters showed tht the cross-section re of cells of the qudriceps muscle of thigh ws smller thn tht of the semimembrnosus muscle. Its increse in the former ws cused, primrily, by the psteuristion process, while in the second one by the injection with brine. The mssging process resulted in more regulr shpes of cells of the qudriceps muscle of thigh thn of those of the semimembrnosus muscle s evidenced by the vlue of the H/V quotient close to 1. Following psteuristion, both the re nd the circumference of the muscle fibres of the m. qudriceps femoris incresed further, while the vlue of H/V in both muscles did not differ sttisticlly significntly despite significnt differences fter mssging. Key words: computer imge nlysis, muscle structure, curing, plsticiztion, psteuristion
2 Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. Introduction Muscle fibres prmeters provide one of the bsic elements of met histologicl structure ssocited with the qulity of finl products mnufctured from whole muscles (AGUILERA 2005). Their size is connected with the ntomicl position of given muscle in the niml crcss. There re differences between individul niml muscles regrding their histologicl structure (WIKLUND et AL. 1998), texture (SHACKELFORD et AL. 1995) s well s in their susceptibility to the process of plsticiztion (MOTYCKA nd BECHTEL 1983, SHACKELFORD et AL. 1989.). Morphologicl prmeters of structurl muscle prmeters chnge under the influence of fter slughter mturtion (MESTRE PRATES et AL. 2002, SOTELO et AL. 2004), s result of technologicl processes to which met is subjected in the course of processing s well s types of chemicl compounds mking up the curing mixture. The technologicl nd technicl curing conditions (LA- CHOWICZ et AL. 2003, SOBCZAK et AL. 2004.) nd therml tretment (PALKA 2004, TORN- BERG 2005) exert significnt impct on the dynmics nd direction of met tissue structure chnges. Brine which is pplied intrmusculrly remins in interfibrillr spces in quntities which depend on the type of the rw mteril (GAJEWSKA-SZCZERBAL nd KRZYWDZIŃSKA-BARTKOWIAK 2005, KRZYWDZIŃSKA-BARTKOWIAK nd GAJEWSKA- -SZCZERBAL 2007). The plsticiztion process by loosening nd destroying muscle structures increses the sorption of the curing brine nd diffusion of intrfibrillr proteins into intrcellulr spces (LACHOWICZ et AL. 2003, XARAGAỳO et AL. 1998). The im of this study ws comprtive ssessment of the histologicl structure of swine hm muscles, nmely: the semimembrnosus (musculus semimembrnosus) nd thigh qudriceps (musculus qudriceps femoris) muscles nd the rnge of its chnges s result of the introduction of the curing brine, plsticiztion nd therml tretment. Mteril nd methods Muscles for nlyses were collected 24 h fter slughter nd divided into two prts, one of which ws treted s the control, while the other the experimentl smple ws injected with curing brine which contined, mong others, 5.0% sodium chloride nd 0.5% polyphosphtes converted into P 2 O 5. The experimentl smples injected with the brine were plsticized in 95% vcuum nd then psteurised in steel cns t the temperture of 72 C in the cn geometric centre mesuring the temperture with the ssistnce of thermocouple sensor. Histologicl preprtions were prepred t ech of the four phses of the experiment stining them with the ssistnce of hemtoxylin nd eosin (HE). Histometric mesurements were performed with the id of the computer imge nlysis (CIA) using for this purpose the MultiScn softwre. The chrcteristion of the microscopic imges ws crried out on the bsis of the following muscle fibres prmeters: cell cross-section re, their circumference nd Feret s H nd V dimeters. The fibres cross-section shpe ws determined on the bsis of the rtio of the Feret s H nd V dimeters. In ddition, the percentge proportion of muscle fibres nd their quntity in the nlysed field of vision were clculted. All the mesurements were crried out three times from 10 fields of vision of the microscope eyepiece. The obtined results were tested for the significnce level of p = 0.05.
Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. 3 Results nd discussion The results of prmeter mesurements of the porcine musculus semimembrnosus nd musculus qudriceps femoris structures re presented in Figures 1-6 s well s in Tble 1. Using visul ssessment system, it ws found tht the scope of chnges depended on the evluted muscle nd the stge of experiment. A B C Fig. 1. Microstructure of the musculus qudriceps femoris: A rw, B injected with brine, C plsticized, D psteurised Rys. 1. Mikrostruktur musculus qudriceps femoris: A surowy, B nstrzyknięty solnką peklującą, C plstyfikowny, D psteryzowny D Differences in the met histologicl structure nd its constitution depending on the niml species nd/or type of muscle were reported erlier by other reserchers, mong others: LIU et AL. (1996), ORYL (2004), GAJOWIECKI et AL. (2001) nd LACHOWICZ et AL. (1997). Anlyzing the results obtined with the ssistnce of the computer imge nlysis it ws found tht the rw semimembrnosus muscle ws chrcterised by greter re nd circumference of muscle fibres nd firly regulr shpe (H/V = 0.97) in reltion to the thigh qudriceps muscle (Tble 1). Similr experiments for rw muscles were conducted erlier nd similr correltion ws obtined (GAJEWSKA-SZCZERBAL et AL. 2007). The process of injection with the curing brine incresed the cell cpcity in the semimembrnosus muscle in comprison with the rw muscle (Figs. 2, 3). This is confirmed by the mesurement results of the cross section re nd circumference of muscle fibres crried out in rw nd injected muscles between which sttisticlly significnt differences were observed (Figs. 3, 4).
4 Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. A B C Fig. 2 Microstructure of the musculus semimembrnosus: A rw, B injected with brine, C plsticized, D psteurised Rys. 2. Mikrostruktur musculus semimembrnosus: A surowy, B nstrzyknięty solnką peklującą, C plstyfikowny, D psteryzowny D Surfce re (μm 2 ) 260 240 220 200 180 160 140 c b b b 120 100 Rw Injected with brine Plsticized Psteurised m. qudriceps femoris m. semimembrnosus Fig. 3. Chnges in the surfce re (CSA) of muscle fibres cused by: the injection with brine, mssging nd psteuristion Rys. 3. Zminy powierzchni przekroju włókien mięśniowych spowodowne przez nstrzyk solnką peklującą, msownie i psteryzcję
Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. 5 Circumference (μm) 260 240 220 200 180 160 140 c b b b 120 100 Rw Injected with brine Plsticized Psteurised m. qudriceps femoris m. semimembrnosus Fig. 4. Chnges in the circumference of muscle fibres cused by: the injection with brine, mssging nd psteuristion Rys. 4. Zminy obwodu włókien mięśniowych spowodowne poprzez nstrzyk solnką peklującą, msownie i psteryzcję Number of muscle cells 80 70 60 50 40 30 20 10 c c b d b bc c b 0 Rw Injected with brine Plsticized Psteurised m. qudriceps femoris m. semimembrnosus Fig. 5. Number of muscle cells in the field of view of the exmined muscles: rw, injected with brine, mssged nd psteurised Rys. 5. Liczb komórek mięśniowych w polu obrzu bdnych mięśni: surowych, nstrzykniętych solnką peklującą, msownych i psteryzownych The incresed re of muscle fibres in the semimembrnosus muscle cused simultneous decline of their numbers in the microscopic field of vision (Fig. 5). The pplied mssging nd psteuristion processes cused shrinkge of fibres nd incresed the mount of muscle cells counted in the field of vision nd the re of the exmined picture occupied by them. In contrst to the semimembrnosus muscle, the injection of the thigh qudriceps muscle with the curing brine cused shrinkge nd decrese of the cross-section re of its cells (Fig. 1). On the other hnd, their numbers, circumference
6 Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. Proportion of muscle cells (%) 85 80 75 70 65 60 55 50 d Rw d d Injected with brine bc c Plsticized b bc Psteurised m. qudriceps femoris m. semimembrnosus Fig. 6. Proportion of muscle cells in the field of view of the exmined muscles: rw, injected with brine, mssged nd psteurised Rys. 6. Zwrtość komórek mięśniowych w polu obrzu bdnych mięśni: surowych, nstrzykniętych solnką peklującą, msownych i psteryzownych Tble 1. The effect of injection, mssging nd psteuristion on chnges in structure prmeters of musculus qudriceps femoris (I) nd musculus semimembrnosus (II) Tbel 1. Wpływ nstrzyku, msowni i psteryzcji n zminy prmetrów struktury musculus qudriceps femoris (I) i musculus semimembrnosus (II) Experiment s phse The kind of muscle Feret s dimeter H (μm) V (μm) H/V Rw muscles I 54.9 b ±4.1 72.7 ±6.9 0.75 II 63.6 bc ±6.9 65.7 bc ±3.7 0.97 Muscles injected with brine I 52.9 ±3.4 59.5 de ±5.2 0.93 II 76.2 d ±12.9 68.4 b ±8.2 1.11 Mssged muscles I 65.2 bc ±2.8 59.5 de ±5.2 1.09 II 68.1 cd ±11.9 62.2 bcde ±6.4 0.91 Psteurised muscles I 65.7 bcd ±1.9 64.0 bcd ±2.4 1.02 II 64.1 bc ±13.2 59.2 cde ±5.8 1.1 The sme letters designte men vlues which do not differ significntly t the level of p 0.05. nd res were found incresed following the plsticiztion process. This fct ppers to indicte tht the penetrtion of the mjority of brine into the cells of the semimembrnosus muscle took plce fter the injection with the curing brine, wheres in the cse of the thigh qudriceps muscle, only during the mssging process. However, the vlues of the bove-mentioned histometric chrcteristics filed to differ sttisticlly significntly in comprison with rw muscles (Figs. 3, 4, 5, 6).
Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. 7 Muscle fibres exposed to het shrink s wter derived both from intrcellulr spces s well s from muscle fibres themselves re liberted. This phenomenon is cused by the denturtion nd cogultion of srcoplsmtic nd myofibrillrs proteins, s well s by reduced cpbility to hold wter immobilized within myofibrils (SHACKELFORD et AL. 1989). In the presented experiments, the pplied psteuristion process consolidted the structure which developed in the course of the plsticiztion process. Conclusions 1. A significnt influence ws determined of the curing brine constituents, s well s of the mechnicl tretments nd therml processing on chnges of the histometric prmeters of the exmined muscle structure. 2. When compring the structure elements of both hm muscles, i.e.: the semimembrnosus (musculus semimembrnosus) nd thigh qudriceps (musculus qudriceps femoris) muscles, it ws found tht the first of them ws chrcterised by higher vlues of the exmined chrcteristics in comprison with the second one. 3. Fibres of the semimembrnosus muscle were more susceptible to the brine bsorption during injection nd plsticiztion processes thn the cells of the thigh qudriceps muscle. References AGUILERA J.M., 2005. Why food microstructure? J. Food Eng. 67: 3-11. GAJEWSKA-SZCZERBAL H., KRZYWDZIŃSKA-BARTKOWIAK M., KORBIK T., 2007. Anlysis of chnges of the histologicl structure of hm muscles s ffected by curing nd therml tretment. Pol. J. Food Nutr. Sci. 57, 2, 227-232. GAJEWSKA-SZCZERBAL H., KRZYWDZIŃSKA-BARTKOWIAK M., 2005. Chnges in histologicl structure of porcine nd bovine semimembrnosus muscles under the influence of mechnicl nd therml procedures. Annu. Anim. Sci. 2: 25-29. GAJOWIECKI L., LACHOWICZ K., ŻYCH A., SOBCZAK M., KOTOWICZ M., ŻOCHOWSKA J., KŁOS B., 2001. A comprtive nlysis of technologicl utility of selected chicken muscles in mssged product mnufcture. Foli Univ. Agric. Stetin. 220, Sci. Alim. 1: 29-34. KOŁCZAK T., 1983. Biologiczne podstwy technologii mięs. Wyd. AR, Krków. KRZYWDZIŃSKA-BARTKOWIAK M., GAJEWSKA-SZCZERBAL H., 2007. Effect of curing by injection method, mssging nd psteuristion on histologicl chnges in bovine muscles. Electr. J. Pol. Agric. Univ. Ser. Food Sci. Technol. 10, 2. LACHOWICZ K., GAJOWIECKI L., KLEMIE A., 1997. Effect of polyphosphte nd soy protein on texture nd rheologicl properties of smoked loin obtined from ple, soft, exudtive (PSE)- -met. Pol. J. Food Nutr. Sci. 6, 47, 4: 93-101. LACHOWICZ K., SOBCZAK M., GAJOWIECKI L., ŻYCH A., 2003. Effects of mssging time on texture, rheologicl properties, nd structure of three pork hm muscles. Met Sci. 63: 225-253. LIU A., NISHIMURA T., TAKAHASHI K., 1996. Reltionship between structurl properties of intrmusculr connective tissue nd toughness of vrious chicken skeletl muscles. Met Sci. 43: 93-96.
8 Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. MESTRE PRATES J.A., GARCIA E COSTA F.J.S., RIBEIRO A.M.R., DIAS CORREIA A.A., 2002. Contribution of mjor structurl chnges in myofibrils to rbbit met tenderiztion during geing. Met Sci. 61: 103-113. MOTYCKA R.R., BECHTEL P.J., 1983. Influence of pre-rigor processing, mechnicl tenderiztion, tumbling method nd processing time on the qulity nd yield of hm. J. Food Sci. 48: 1532- -1536. ORYL B., 2004. Structure nd hrdness of selected muscles in young bulls. Pol. J. Food Nutr. Sci. 13/54, 1: 57-63. PALKA K., 2004. The influence of post-mortem geing nd rosting on the microstructure, texture nd collgen solubility of bovine semitendinosus muscle. Met Sci. 68: 191-198. SHACKELFORD S.D., REAGAN J.O., MANN T.F., LYON C.E., MILLER M.F., 1989. Effect of blde tenderiztion, vcuum mssge time nd slt level on chemicl, texturl nd sensory chrcteristics of precooked chuck rost. J. Food Sci. 54: 843-845. SHACKELFORD S.D.,WHEELER T.L., KOHMARAIE M., 1995. Reltionship between sher force nd trined sensory pnel tenderness rtings of 10 mjor muscles from Bos indicus nd Bos turus cttle. J. Anim. Sci. 73: 3333-3340. SOBCZAK M., LACHOWICZ K., CZARNECKI R., GAJOWIECKI L., KLEMKE A., ŻOCHOWSKA J., 2004. Comprtive nlysis of the susceptibility of selected muscles of pietrin, duroc nd Polish lrge white Polish lndrce pigs to mssge-induced chnges. Pol. J. Food Nutr. Sci. 13/54, 2: 179-184. SOTELO I., PÉREZ-MUNUERA I., QUILES I., HERNANDO I., LARREA V., LLUCH M.A., 2004. Microstructurl chnges in rbbit met wrpped with Pteridium guilinum fern during postmortem storge. Met Sci. 66: 823-829. TORNBERG E., 2005. Effects of het on met proteins-implictions on structure nd qulity of met products. Rev. Met Sci. 70: 493-508. WIKLUND E., MALMFORS G., LUNGSTRÖM K., 1998. The effects of exercise on muscles fibre composition nd oxidtive cpcity in eight bovine skeletl muscles. Swed. J. Agric. Res. 28: 111- -116. XARAGAỳO M., FREIXANTE L., LAGARES J., FERNANDEZ E., DE JAGER-PONET P., 1998. Wirkung der Vormssge bei der Herstellung gegrter Fleischerzeugnisse us gnzen Muskeln. Fleischwirtschft 78: 953-995. ZMIANY PARAMETRÓW STRUKTURY MIĘSA WIEPRZOWEGO PODCZAS PEKLOWANIA I PASTERYZACJI Streszczenie. Celem prcy był porównwcz ocen struktury histologicznej świńskich mięśni szynkowych: półbłonistego (musculus semimembrnosus) i czterogłowego ud (musculus qudriceps femoris) orz zkresu jej zmin podczs nstrzyku solnką peklującą pod ciśnieniem 0,5 MP, plstyfikcji w tmosferze próżni orz psteryzcji. Mięśnie pobierno z tusz po 24 h od uboju. W skłd solnki nstrzykowej wchodziły m.in. chlorek sodu i wielofosforny. W celu wykonni pomirów histometrycznych w kżdej fzie doświdczeni sporządzno preprty histologiczne, wybrwione metodą HE (hemtoksyliną i eozyną). Pomiry wykonywno z pomocą komputerowej nlizy obrzu (KAO), stosując progrm MultiScn. Obrzy mikroskopowe schrkteryzowno n podstwie nstępujących prmetrów włókien mięśniowych: powierzchni przekroju poprzecznego komórek, ich obwód orz średnice Feret H i V. Oprócz tego obliczono procentowy udził włókien mięśniowych orz ich liczbę w nlizownym polu widzeni. Ze stosunku średnic Feret H i V określono ksztłt przekroju włókien. Wyniki wszystkich pomirów
Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127. 9 testowno dl poziomu istotności p = 0,05. Anliz porównwcz prmetrów struktury wykzł, że powierzchni przekroju komórek mięśni czterogłowego ud był mniejsz niż półbłonistego. N jej zwiększenie w pierwszym z mięśni wpłynął przede wszystkim proces psteryzcji, ntomist w drugim nstrzyk solnką. Proces msowni spowodowł uzysknie brdziej regulrnych ksztłtów komórek mięśni czterogłowego ud niż w mięśniu półbłonistym, czego wyrzem był wrtość ilorzu H/V zbliżon do 1. Po psteryzcji nstąpiło dlsze zwiększenie powierzchni i obwodu włókien mięśniowych m. qudriceps femoris, wrtości H/V w obu mięśnich nie różniły się sttystycznie mimo istotnych różnic po msowniu. Słow kluczowe: komputerow nliz obrzu, struktur mięśni, peklownie, plstyfikcj, psteryzcj Corresponding ddress Adres do korespondencji: Hnn Gjewsk-Szczerbl, Instytut Technologii Mięs, Uniwersytet Przyrodniczy w Poznniu, ul. Wojsk Polskiego 31/33, 60-624 Poznń, Polnd, e-mil: hgszcz@up.poznn.pl Accepted for print Zkceptowno do druku: 7.10.2009 For cittion Do cytowni: Gjewsk-Szczerbl H., Krzywdzińsk-Brtkowik M., Jrmołowsk-Jurczyszyn D., 2009. Chnges in pork muscles structure prmeters in the course of curing nd psteuristion. Nuk Przyr. Technol. 3, 4, #127.