Quality and Shelf Life of the Adductor Muscle of Lion’s Paw Scallop Nodipecten subnodosus Transported and Stored Whole in Refrigeration

Edgar I. Jiménez-Ruiz, Víctor M. Ocaño-Higuera, Alfonso N. Maeda-Martínez, Francisco J. Castillo-Yáñez, Alejandro Varela-Romero, Abril Z. Graciano-Verdugo and Enrique Márquez-Ríos

SUMMARY

The effect of refrigerated transport during 48h and four days ing to this parameter the product was considered edible until the storage on the quality and shelf life of the adductor muscle of end of the storage period. With respect to TVB-N and TMA-N, whole lion’s paw scallops (Nodipecten subnodosus) was evalu- the allowed limits were not exceeded. The pH level showed no ated. Proximal composition, ATP and related products, K-value, significant variations during storage. According to TVB-N, TMA- total volatile bases (TVB-N) and trimethylamine (TMA-N), pH, N values and microbiological analyses performed, adductor and microbiological analyses were quantified. The muscle was muscle proved to be innocuous after four days under the condi- found to be lean and high in protein. Levels of muscular ATP tions of transportation and storage utilized. were initially low and the K-value increased lineally, but accord-

Introduction species is its high growth in their shells, but are also scallop, maximize its value, rate, as it attains a size of served raw in dishes of the and promote its consumption, The lion’s paw scallop No- 7cm (commercial size) in only sushi or sashimi type (Paust is the quality and innocuous dipecten subnodosus consti­ eight months. For the afore­ and Rice, 2001; Pacheco- nature of the final product. tutes one of the most impor­ mentioned reasons, fish farm­ Agui­lar et al., 2001). Like many other aquatic or­ tant fishery resources on ers have become interested in In terms of the nutritional ganisms, upon their death the Mexico’s Baja California Pe­ producing this species and value of pectinids, the main pectinids undergo a series of ninsula. The commercial im­ significant efforts are under­ edible portion is the adductor biochemical changes that portance of this scallop rests way to consolidate its cultiva­ muscle, which has a high pro­ strongly impact the quality on the size of the organisms tion (Massó-Rojas et al., tein content and a high bio­ and shelf life of finished (22cm tall) and their weight 2001; Pacheco-Aguilar et al., logical value, comparable to products. These postmortem (250g at 5 years’ growth), the 2001; Maldonado-Amparo et that of casein and meat. changes can be studied by price (USD16/kg in the inter­ al., 2004). Moreover, it is classified as monitoring certain parameters national market), flavor, and While in both Mexico and lean food due to the fact that and indicators of quality and the production of their adduc­ the USA the principal product its lipid content is ≤0.7% freshness (Sikorski et al., tor muscle. Its production is consumed is the fresh refrig­ (Oca­ño-Higuera et al., 2001). 1990; Abbas et al., 2008; concentrated in the Ojo de erated adductor muscle, other In addition to the nutrition­ Oca­­ño-Higuera et al., 2009; Liebre Lagoon, state of Baja countries, mainly in Asia and al aspects, another factor that 2011; Liu et al., 2010). California Sur (BCS), and in Europe, have a high demand must be considered in the At present there is no fea­ recent years it has stabilized for the whole shelled bivalves. seafood product market so as sible and established strategy at 100ton/year. Another im­ These products are usually to determine the optimum for exporting and distributing portant characteristic of this consumed after being cooked means of distributing this this particular pectinid in its

KEYWORDS / Lion’s Paw Scallops / Muscle / Quality / Shelf Life / Received: 09/01/2011. Modified: 05/20/2012. Accepted: 05/23/2012.

Edgar Iván Jiménez Ruiz. Bio­ sor, UNISON, Mexico. Ad­ Francisco Javier Castillo Yá­ gy of Aquatic Organisms, chemical Engineer, Instituto dress: Departamento de Cien­ ñez. Degree in Chemical Biol­ CIAD, Mexico. Professor, Tecnológico de Los Mochis, cias Químico Biológicas, Uni­ ogy, Uson, Mexico. M.Sc., In­ UNISON, Mexico. Mexico. M.Sc., Centro de In­ versidad de Sonora. Encinas y stituto Tecnológico de Guay­ Abril Zoraida Graciano Ver­ vestigación en Alimentación y Rosales s/n, Hermosillo, So­ mas, Mexico. Ph.D., CIAD, dugo. Degree in Chemical Bi­ Desarrollo A.C. (CIAD), Mex­ nora 83000, Mexico. e-mail: México. Professor, UNISON, ology, Uson, Mexico. M.Sc. ico. Ph.D. candidate, Universi­ [email protected] Mexico. and Ph.D., CIAD, Mexico. dad de Sonora (UNISON), Alfonso Nivardo Maeda Mar­ Alejandro Varela Romero. Ma­ Professor, UNISON, Mexico. Mexico. tínez. Ph.D., University of rine Biologist, Universidad Enrique Márquez Ríos. Degree Victor Manuel Ocaño Higuera. Southampton, RU. Researcher, Autónoma de Baja California in Chemical Biology, Uson, Degree in Chemical Biology, Centro de Investigaciones Bi­ Sur, Mexico. M.Sc. in Popula­ Mexico. M.Sc. and Ph.D., Uson, Mexico. M.Sc. and ológicas del Noroeste, S.C. tion Genetics, UNISON, Mex­ CIAD, Mexico. Professor, Ph.D., CIAD, Mexico. Profes­ (CIBNOR), Mexico. ico. Ph.D. in Molecular Biolo­ UNISON, Mexico.

464 0378-1844/12/06/464-06 $ 3.00/0 JUN 2012, VOL. 37 Nº 6 CALIDAD Y VIDA DE anaquel del mÚsculo aBductor deL PECTÍNIDO Nodipecten subnodosus TRANSPORTADO Y ALMACENADO ENTERO BAJO REFRIGERACIÓN Edgar I. Jiménez-Ruiz, Víctor M. Ocaño-Higuera, Alfonso N. Maeda-Martínez, Francisco J. Castillo-Yáñez, Alejandro Varela-Romero, Abril Z. Graciano-Verdugo y Enrique Márquez-Ríos RESUMEN

Se evaluó el efecto del transporte bajo refrigeración durante tó linealmente, aunque de acuerdo a este parámetro el producto 48h y almacenaje por cuatro días sobre la calidad y vida de ana- se consideró comestible hasta el final del periodo de almacena- quel del péctinido mano de león (Nodipecten subnodosus). Se miento. Los límites permitidos de BVT-N y TMA-N no fueron ex- cuantificó la composición proximal, el ATP y compuestos relacio- cedidos. El pH no mostró variaciones significativas durante el nados, el valor K, bases volátiles totales (BVT-N) y trimetilamina almacenamiento. De acuerdo a los valores de BVT-N y TMA-N (TMA-N) y el pH, y se realizaron análisis microbiológicos. Se obtenidos y a los resultados de los análisis microbiológicos, el encontró que el músculo era magro y de alto contenido proteico. músculo abductor probó ser inocuo tras cuatro días bajo las con- Los niveles de ATP fueron inicialmente bajos y el valor K aumen- diciones de transporte y almacenaje utilizadas.

QUALIDADE E VIDA EM PRATELEIRA dO mÚsculo adutor dO PECTÍNIDO Nodipecten subnodosus TRANSPORTADO E ARAMAZENADO INTEIRO SOB-REFRIGERAÇÃO Edgar I. Jiménez-Ruiz, Víctor M. Ocaño-Higuera, Alfonso N. Maeda-Martínez, Francisco J. Castillo-Yáñez, Alejandro Varela-Romero, Abril Z. Graciano-Verdugo e Enrique Márquez-Ríos RESUMO

Avaliou-se o efeito do transporte sob-refrigeração durante 48h aumentou linearmente, mesmo que de acordo a este parâmetro e armazenagem por quatro dias sobre a qualidade e vida em o produto se considerou comestível até o final do período de prateleira da vieira pata-de-leão (Nodipecten subnodosus). armazenamento. Os limites permitidos de BVT-N e TMA-N não Quantificou-se a composição proximal, o ATP e compostos re- foram excedidos. O pH não mostrou variações significativas du- lacionados, o valor K, bases voláteis totais (BVT-N) e trimetila- rante o armazenamento. De acordo aos valores de BVT-N e mina (TMA-N) e o pH, e se realizaram análises microbiológicas. TMA-N obtidos e aos resultados das análises microbiológicos, Encontrou-se que o músculo era magro e de alto conteúdo pro- o músculo adutor provou ser inócuo após quatro dias sob as téico. Os níveis de ATP foram inicialmente baixos e o valor K condições de transporte e armazenagem utilizadas.

whole, fresh refrigerated, the adductor muscle of whole then cleared in 200 liter fi­ ing the coolers, 5kg of frozen presentation for consumption lion’s paw scallops during berglass tanks for 1h. refrigerant gel (Dolphin Blue on the half shell, as is the refrigerated transport and Crabs) at -20ºC was intro­ case with oysters. In prac­ storage, as a possible alterna­ Experiment on transport and duced and a thermograph was tice, handlers strive to main­ tive for exporting the product storage placed to monitor the internal tain the cold chain right from to international markets. temperature of the coolers the moment when the speci­ The packing of the organ­ during transport, as shown in mens are harvested in their Materials and Methods isms prior to transport in­ Figure 1. The refrigerant gel natural habitat until they are volved placing them in cool­ was separated from the upper processed and/or consumed, Experimental organisms ers made of expanded poly­ layer of scallops by another but this is not always styrene with a capacity of 45 bed of moistened brown pa­ achieved. It is also worth to Adult organisms of lion’s liters. The live organisms per. At that moment, the ad­ note that when delivered paw scallops, N. subnodosus, were arranged dorso-ventrally ductor muscles of six scallops fresh to intermediaries or fi­ with a height of ~10cm were with elastic bands attached to were frozen in liquid N2, to nal sellers, whole organisms utilized, after being harvest­ prevent them from opening be used to determine the ini­ can fetch a price as high as ed from a culture system lo­ their valves and possibly dry­ tial biochemical conditions at 2.00 USD/scallop. It is thus cated in Bahía Tortugas, ing out. A total of 120 organ­ the time of transport. necessary to look for and BCS, in June 2009. Before isms were collected and dis­ Transportation was simu­ design alternative storage transport and storage the tributed into two coolers. lated under the most realistic methods during transport and scallops were cleaned with During the packing process, conditions possible. In the distribution that will assure an oyster machine that func­ the scallops were placed in a first phase, the coolers with that the quality and adequacy tions on the basis of sprayers horizontal position and a bed the refrigerant gel and the of the product will be main­ and brushes (Ets. Bertrand, of brown paper moistened scallops were carried in a tained until offered to the fi­ Marennes, France). The with seawater was placed be­ pick-up truck at ambient nal consumer. The objective cleaning process was com­ tween the layers of mollusks. temperature for 14h. Then a of this study was to evaluate pleted with the specimens Once the organisms were second stage of transporta­ the quality and shelf life of being brushed by hand and packed, and just before seal­ tion was simulated, in which

JUN 2012, VOL. 37 Nº 6 465 the sealed coolers were TMA-N placed in a refrigerated room at the installations The determination of TMA- of CIBNOR, La Paz, N was also conducted follow­ BCS. This procedure was ing the method described by meant to simulate the sec­ Woyewoda et al. (1986) with ond stage of transporta­ only slight modifications of tion, as when the organ­ the weights and volumes used isms are transported in in the extraction. The adduc­ refrigerated trucks. The tor muscle (3g) was homoge­ total simulated transporta­ nized in 5ml of 7.5% trichlo­ tion time was 48h. This Figure 1. Preservation of organisms in refrigeration conditions, indicating roacetic acid in a T18 Basic simulation of the stages of temperature during the process. Ultra-turrax (IKA Works). transportation was de­ The homogenate was centri­ signed in accordance with fuged at 3200g (4ºC) in a the conditions assumed nec­ in order to determine wheth­ to separate the compounds. model IEC-MULTI RF Ther­ essary to deliver this product er or not the parts were apt The mobile phase used was a mo Electron refrigerated cen­ to an international market, for human consumption. In phosphate buffer made up of trifuge (Thermo Fisher Scien­ in this case that of the USA. this procedure, all of the 0.04M KH2PO4 and 0.06M tific, Asheville, NC, USA) for In the last phase, the organ­ organisms’ moist tissues K2HPO4. The flow rate uti­ 15min, and the supernatant isms were removed from the were extracted and homoge­ lized was 1ml·min‑1, and de­ then filtered through What­ coolers and placed in refrig­ nized together with the in­ tection was carried out at man Nº 4 paper and stored at erated storage (3-6ºC) in tervalvular liquid. 254nm in a UV-Vis Varian -80ºC until analysis. polyethylene bags to simu­ Prostar 325 detector. The val­ For quantification, 1ml of late the actual storage facili­ Proximal analysis ues obtained were expressed the filtrate was taken and ties utilized by the final re­ as μmol·g‑1 of sample. The transferred to a screw cap test ceiver, who would then un­ Quantification of moisture, K-value (%) was calculated tube, diluted to 2ml with dis­ dertake to sell the product. proteins, lipids, carbohydrates according to the equation de­ tilled water, and 0.5ml of 10% At that moment, the adduc­ and ash content of the adduc­ scribed by Saito et al. (1959): formaldehyde was added. tor muscles of six organisms tor muscle were carried out Next, 1.5ml of 25% KOH and were dissected and frozen in following the methodologies K-value (%)= 5ml dry toluene were added. liquid N2 for later determi­ described in AOAC (2005). ((HxR+Hx)/(ATP+ADP The tubes were shaken for nation of the effects of +AMP+IMP+HxR+Hx))×100 15min in a Vortex G-560 agi­ transportation and to estab­ pH tator (Scientific Industries lish the quality and shelf life where ATP: adenosine 5′tri­ Inc., Bohemia, NY, USA) and at day 0. Finally, a thermo­ The determination of pH phosphate, ADP: adenosine a pipette was used to remove graph was placed to monitor was conducted following 5′diphosphate, AMP: adenos­ 3.5ml of the toluene layer to the temperature of the re­ Woyewoda et al. (1986), ine 5′monophosphate, IMP: be transferred to another test frigerated room throughout where a homogenate of the inosine 5′monophosphate, tube, this one with 0.3g of product storage (Figure 1). adductor muscle was prepared HxR: inosine, and Hx: hypo­ anhydrous Na2SO4. The mix­ with distilled water and pH xanthine. ture was shaken again until it Sampling for the study of was then measured using an turned transparent. Then, refrigeration Orion 420 potentiometer (Ter­ TVB-N 2.5ml of this solution were mo Electron Corp.), calibrated taken and added to a test tube The organisms transported daily using pH 4 and 7 stan­ The determination of TVB- containing 2.5ml of a solution under the simulated condi­ dards as references. N was conducted following of 0.02% picric acid in dry tions described above re­ the method described by toluene, which was then shak­ mained in the refrigerated ATP, its breakdown products, Woyewoda et al. (1986). Here, en to obtain the final solution. storage area for a period of and the K-value 2g of the sample were taken Measurements were conducted four days, which is a com­ and homogenized in a distill­ in a Varian UV-Visible Cary mon time to market these The extraction, identifica­ ing flask with 300ml water 100 Bio spectrophotometer products. During this time, tion, and quantification of the and 2g MgO, using a T18 Ba­ (Varian Australia Pty Ltd.) at the adductor muscles of six ATP (adenosine 5′triphos­ sic Ultra-turrax (IKA Works, 410nm, using a standard organisms were collected, phate) and the products of its Inc, Wilmington, NC, USA). curve in the range of 0.1- dissected, and frozen in liq­ breakdown in the adductor Twenty drops of vegetable oil 10mg of TMA-N. Results uid N2 every 24h, and then muscle were conducted were added as anti-foaming were expressed as mg TMA- stored at -80ºC until analy­ through high resolution liquid agent and the solution was N/100g of muscle. sis. At each point of the chromatography. The method then distilled for 25min. The sampling program pH, ATP described by Ryder (1985) distillate was collected in Microbiological analyses and related products, K-val­ was followed, where 20μl of 15ml of 2% boric acid and ue, total volatile bases the diluted extract were in­ then titrated with 0.05N Microbiological analyses (TVB-N) and trimethylamine jected into a Varian Prostar H2SO4. A control sample was were carried out according to (TMA-N) were monitored. 240 chromatograph (Varian distilled under the same con­ the recommended methods set Similarly, throughout the Inc., Lake Forest, CA, USA) ditions and the values ob­ out in NOM-031-SSA1-1993 storage period, microbiologi­ using a 4.6×150mm C18 re­ tained were expressed as mg (NOM, 1995) every 24h for 4 cal analyses were conducted versed phase column (Varian) TVB-N/100g of sample. days. A total of six organisms

466 JUN 2012, VOL. 37 Nº 6 were collected for each 24h TABLE I period. The moist tissues VALUES OF pH, NUCLEOTIDES, NUCLEOSIDES AND BASES (µmol·g-1), BVT-N were extracted from the scal­ AND TMA-N (mg/100) OF THE LION’S PAW SCALLOP N. subnodosus ADDUCTOR MUSCLE lops and homogenized to ob­ TRANSPORTED AND STORED WHOLE IN REFRIGERATION CONDITIONS (3-6ºC) tain a representative sample. These procedures were con­ Storage day Parameter Initial Pos-transport ducted at the CIBNOR Micro­ organisms (day 0) biological Diagnostics Labora­ 1 2 3 4 tory in La Paz, BCS, where pH 7.08 ±0.02 a 6.43 ±0.05 b 6.29 ±0.10 c 6.29 ±0.06 c 6.27 ±0.04 c 6.30 ±0.09 c the number of aerobic bacte­ ATP 0.03 ±0.00 bd 0.02 ±0.00 cde 0.05 ±0.01 a 0.04 ±0.02 abe 0.04 ±0.01 ab 0.05 ±0.02 ab ria and fecal coliforms were ADP 0.33 ±0.01 a 0.25 ±0.03 b 0.26 ±0.08 b 0.23 ±0.05 b 0.21 ±0.01 b 0.18 ±0.02 b determined in tryptone yeast AMP 3.95 ±0.37 a 3.61 ±0.68 a 1.44 ±0.46 b 1.08 ±0.39 b 0.99 ±0.31 b 0.80 ±0.05 b extract agar media (48h at 35 IMP 0.05 ±0.00 b 0.08 ±0.01 a 0.12 ±0.05 a 0.15 ±0.07 a 0.13 ±0.02 a 0.14 ±0.04 a ±2ºC) and EC media (44 HxR 0.10 ±0.02 e 0.76 ±0.1 d 1.43 ±0.30 c 2.00 ±0.39 ab 2.04 ±0.30 a 1.59 ±0.05 bc ±2ºC), respectively. Also, Hx 0.12 ±0.03 c 0.10 ±0.09 c 0.37 ±0.07 b 0.49 ±0.07 ab 0.57 ±0.16 a 0.58 ±0.10 a samples were taken to be ana­ TBV-N 6.19 ±1.62 b 6.97 ±2.35 b 11.66 ±1.76 a 12.45 ±2.23 a 12.44 ±1.93 a 13.89 ±2.01 a lyzed for Vibrio cholerae and TMA-N 0.16 ±0.00 c 0.19 ±0.01 a 0.17 ±0.01b c 0.17 ±0.01 bc 0.17 ±0.01 b 0.17 ±0.01 b Salmonella spp. (six whole Values are mean and standard deviation of n= 6. organisms) for days 1 and 4 Means in the same row with different superscript letters are statistically different (P<0.05). of refrigerated storage. The V. cholerae and Salmonella spp. tests were carried out in TCBS media (24h at 35-37ºC) 1.40 ±0.10 and, as was the (Durán et al., 2008; Mørkøre and while stored whole in re­ and brilliant green (24h at 35 case for the values of the rest et al., 2008). frigeration. The first organ­ ±0.5ºC), respectively. of the components, similar to The initial neutral pH values isms obtained immediately that reported by Beltrán-Lugo recorded in the recently col­ after the cleaning process and Statistical analysis et al. (2006) and Pacheco- lected organisms diminished those collected after the 48h Aguilar et al. (2008) for N. significantly (P<0.05) during period of simulated transport A one-way variance analy­ subnodosus and by Ocaño- the stage of simulated trans­ showed low levels of muscu­ sis (ANOVA) was conducted. Higuera et al. (2006) for the port and on the first day of lar ATP, values that may be When significant differences catarina scallop Argopecten refrigerated storage. However, the result of the stress gener­ were found, the multiple com­ ventricosus. after that day and up to the ated during the conditioning parison Tukey test was ap­ end of the study, no further and cleaning of the organ­ plied using the NCSS Ver. pH changes were observed. Ac­ isms, which fostered the 2000 program (Hintze, 2001). cording to Riaz and Qadri breakdown of this energy me­ Also, in some cases regres­ Table I presents the data on (1985), an increase >0.1 units tabolite. The concentration of sion analyses were run. A the pH of the adductor muscle during storage indicates dete­ this metabolite increased on significance level of 5% was of the lion’s paw scallop rioration and loss of freshness, the first day (P<0.05), but set for all of the analyses con­ transported in immersion and as occurs when alkaline-type then remained with no sig­ ducted. stored whole under refrigera­ metabolites are produced as a nificant variation during the tion. The initial value of this result of bacterial proliferation. rest of the storage period Results and Discussion parameter in the organisms In the present study, the pH of (P>0.05). This increase on immediately after collection the adductor muscle did not day 1 may be due to the re­ Proximal analysis and before conditioning and increase during storage, so it generation of ATP through storage (pH= 7.02) is similar can be said that in relation to the action of the adenylate The percentage of moisture to that reported by Kimura et this parameter the adductor kinase enzyme, or to the uti­ in the adductor muscle was al. (1999) for P. yessoensis, muscle retained its edible lization of the phosphoargi­ 75.18 ±0.94 and was charac­ but higher than the value of quality throughout the period nine phosphagen in a reaction terized by the presence of a 6.8 reported by Beltrán-Lugo and under the conditions used catalyzed by the arginine ki­ high protein content of 17.88 et al. (2006), also for N. in this study. This behavior nase enzyme, as has been re­ ±0.46% and a low lipid con­ subnodosus. The difference and similar values have been ported for other species, such tent of 0.11 ±0.01%. For between the latter figure and reported for N. subnodosus as the abalone Haliotis discus this reason, and following the initial value found in the and for A. ventricosus by Pa­ (Watanabe et al., 1992) and the classification reported present study may be due to checo-Aguilar et al. (2008) the scallop Zygochlamys pata- by Domínguez and Gutiér­ the conditions employed by and by Ocaño-Higuera et al. gonica (Massa et al., 2001). rez (1993), the muscle was Beltrán-Lugo et al. (2006) (2006), respectively. The concentration of muscu­ considered lean (<5% lip­ during transport to the site lar ADP showed a significant ids). As is the case with where analyses were conduct­ ATP, Products of its variation during transport most pectinids, N. subnodo- ed, as the specimens spent Breakdown, and the K-value (P<0.05), but no important sus also contains consider­ 30min in a simple cooler con­ variations during the storage able amounts of carbohy­ ditioned with seawater. Under Table 1 shows the values (P<0.05), whereas AMP di­ drates, mostly glycogen, that those conditions, a first stage for ATP and related products minished significantly (P<0.05) serve as an energy reserve in of anaerobic glycolysis may obtained in the adductor mus­ after day 1 of storage, but then the adductor muscle; the val­ have taken place, with the cle of the lion’s paw scallop became stable at ~1μmol·g‑1, ue was 7.08 ±1.58%. Finally, concomitant lactic acid pro­ at the moment of collection, and IMP was found in concen­ the percentage of ash was duction and pH reduction during transport in emersion, trations ≤0.15μmol·g‑1 through­

JUN 2012, VOL. 37 Nº 6 467 uct remained edible until establishes a maximum limit the end of the storage pe­ of 30mg/100g. According to riod. In a regression anal­ this parameter and our analy­ ysis, the K-value of the sis, the adductor muscle was adductor muscle during considered apt for human con­ the refrigerated storage sumption at the end of the phase showed a linear in­ study in refrigeration. crease (y= 12.07x +27.04, r 2= 0.90, P<0.05). This TMA-N behavior was reported pre­ viously by Ocaño-Higuera One indicator that is com­ Figure 2. K-value in adductor muscle of the lion’s paw scallop N. subnodosus stored whole in refrigeration conditions (3-6ºC). Data shown are mean of n=6. et al. (2006) for A. ventri- monly used to evaluate bacte­ Bars represent standard deviation. cosus. rial deterioration in seafood products is the determination TVB-N of TMA-N, a compound with out the experiment. For the rectly from the culture site, the a characteristic ammonia odor latter two compounds, which value obtained was low, 4.58 Determining TVB-N al­ that may reflect bacterial dete­ are related to the sweet, pleas­ ±0.41% (data not shown on the lowed to quantify the low mo­ rioration (Ryder et al., 1984). ant flavor of seafood products, graph), although it increased lecular weight volatile bases As the results obtained from a behavior similar to that re­ significantly (P<0.05) during and amino compounds pro­ the muscle shown in Table I ported in P. yessoensis (Ka­ simulated transport, reaching a duced by the microbial decar­ reveal, the initial values of washima and Yamanaka, value of 19.77 ±7.11% on day 0 boxylation of amino acids and TMA-N of organisms obtained 1992), Z. patagonica (Massa et of the refrigerated storage ex­ nitrogenated compounds, directly from the lagoon in­ al., 2001), A. ventricosus (Oca­ periment. This is due to the which are widely used as indi­ creased during the transport ño-Higuera et al., 2006), and increased HxR concentration cators of shelf life and, more stage, and variations were also N. subnodosus (Pacheco-Agui­ at this time. specifically, to measure stages observed during storage lar et al., 2008) was observed. Saito et al. (1959) and of deterioration or loss of (P<0.05). However, the TMA- On the other hand, HxR and Sikorski et al. (1990) elabo­ freshness. Table I presents the N concentrations found for the Hx showed significant varia­ rated a rather general classifi­ TVB-N values for the adduc­ muscle under the conditions of tions during the experiment, cation of the quality of sea­ tor muscle of lion’s paw scal­ transport and storage used but their concentrations during food products with respect to lop N. subnodosus transported were lower than those report­ the storage period were always their K-value. In it, a K-value in immersion and stored whole ed in other studies conducted below those reported by Oca­ <20% indicates a ‘very fresh’ in refrigerated conditions. It with this same species and ño-Higuera et al. (2006) and product, while products with shows the initial low values of with A. ventricosus, even Pacheco-Aguilar et al. (2008) values between 20 and 50% TVB-N (6.97 ±2.35mg/100g), though the latter included for A. ventricosus and N. are considered ‘moderately which are below those found only adductor muscles stored subnodosus, respectively. This fresh’, and those >70% are in this same species by Pache­ at a temperature of 0°C (Oca­ finding is relevant, since these ‘not fresh’. In terms of this co-Aguilar et al. (2008) and ño-Higuera et al., 2006; Pa­ compounds have been related classification, the object of this those obtained by Ocaño- checo-Aguilar et al., 2008). to a bitter taste in seafood study can be considered ‘very Higuera et al. (2006) for A. This indicator has been asso­ products. The concentrations fresh’ after two days of simu­ ventricosus. In the present ciated with unpleasant odors found in this experiment could lated transport and, indeed, study, an increase (P<0.05) of in fish and other seafood spe­ be the result of low autolytic maintained that standard until TVB-N was seen on day 1 of cies when it exceeds certain and bacterial activity in the the first day of refrigerated storage in refrigeration, but no limits. In this regard, limits adductor muscle. storage, when it reached 44.87 additional significant change of 5-10mg/100g of muscle In the case of the K-value ±3.24%. After that, and de­ was observed (P<0.05). In this have been established (Sikor­ for the adductor muscle (Fig­ spite the significant increase in regard, it is important to point ski et al., 1990), which are ure 2), as was to be expected the K-value (P<0.05), accord­ out that the NOM-029- higher than those found in the in the organisms analyzed di­ ing to this parameter the prod­ SSA1-1993 norm (NOM, 1994) present study.

TABLE II Microbiological Analyses MICROBIOLOGICAL ANALYSIS OF THE TOTAL WET BIOMASS IN THE LION’S PAW SCALLOP N. subnodosus TRANSPORTED AND STORED The results of the microbio­ WHOLE IN REFRIGERATION CONDITIONS (3-6ºC) logical analyses carried out on lion’s paw scallop during re­ Storage time Aerobic mesophiles Faecal coliforms frigerated storage, shown in (MPN/100 g)** Vibrio cholerae Salmonella spp (days) (log10 CFU/g)* Table II, reveal differences in 0 2.42 ±0.08a < 30 ND ND the amount of mesophilic aero­ 1 3.16 ±0.02b < 30 Absent Absent bic bacteria on different days 2 4.32 ±0.09d 40 ND ND of storage (P<0.05). These in­ 3 2.24 ±0.09a 40 ND ND dexes did not reach at any mo­ 4 3.49 ±0.06c 40 Absent Absent ment of the experiment the maximal limit of 5×105 CFU/g *Values are mean and standard deviation of n=2. Means in the same column with different superscript letters are statistically different (P<0.05). (5.69 log10 CFU/g) allowed for **Statistical values calculated from Table 4 of the NMX-AA-042-1987 (NMX, 1987). human consumption according ND= Not Determined. to the NOM-031-SSA1-1993

468 JUN 2012, VOL. 37 Nº 6 norm (NOM, 1994). The high­ CIBNOR and Marimex del Massa AE, Paredi ME, Crupkin M Canizales-Rodríguez DF, Cas­ est values, of 22892 CFU/g Pacífico SA de CV for their (2001) Nucleotide catabolism in tillo-Yáñez FJ, Ruiz-Bustos E, cold stored addcutor muscle of Graciano-Verdugo AZ, Plas­ (4.32 log10 CFU/g) were found support. This work is part of scallop (Zygochlamys patagoni- cencia-Jatomea M (2011) Fresh­ on day 2 of sampling. As in the Ph.D. thesis of the first au­ ca). J. Food Biochem. 26: 295- ness assessment of ray fish the case of aerobic mesophiles, thor at the Universidad de So­ 305. stored in ice by biochemical, the fecal coliforms bacteria did nora, Mexico. Massó-Rojas JA, Morales-Bojórquez chemical and physical methods. not reach in any of the sam­ E, Talavera-Mayer J, Fajardo- Food Chem. 125: 49-54. pling days the maximum limit References León M, Hernández-Valenzuela Pacheco-Aguilar R, Ocaño-Higuera (230 MPN/100g), indicated by R (2001) Almeja mano de león. VM, Maeda-Martínez AN In Cisneros-Mata MA, Belén­ (2001) Manejo y procesado de the norm. The highest values, AOAC (2005) Official Methods of th dez-Moreno, LF, Zárate-Becerra pectínidos. In Maeda-Martínez 40 MPN/100g, were found as Analysis of the AOAC, 18 ed. E, Gaspar-Dillanes MT, López- Association of Official Analyti­ AN (Ed.) Los Moluscos Pec- of day 3, but these levels were González L, Saucedo-Ruiz L, tínidos de Iberómerica: Ciencia cal Chemists, Washington, DC, Tovar-Ávila J (Eds.) Sustentabi- maintained until the end of the USA. 2590 pp. y Acuicultura. Limusa. México. lidad y Pesca Responsable en pp. 431-450. experiment. The foregoing Abbas KA, Mohamed A, Jamilah B, México. Evaluación y Manejo. Pacheco-Aguilar R, Márquez-Ríos demonstrates the innocuous Ebrahimian M (2008) A review 1999-2000. Instituto Nacional de E, Lugo-Sánchez ME, García- nature of the product and the on correlations between fish la Pesca. México. pp. 349-366. freshness and pH during cold Sánchez G, Maeda-Martínez adequacy of the conditions em­ Mørkøre T, Pablo I, Mazo T, Vil­ AN, Ocaño-Higuera VM (2008) storage. Am. J. Biochem. Bio- dana T, Einen O (2008) Impact ployed in the handling of the technol. 4: 416-421. Postmortem changes in the ad­ organisms throughout the ex­ of starvation and handling ductor muscle of Pacific lions- periment. Beltrán-Lugo AI, Maeda-Martínez stress on rigor development paw scallop (Nodipecten subno- AN, Pacheco-Aguilar R, Nolas­ and quality of Atlantic salmon dosus) during ice storage. J. Other important bacteria that co-Soria HG (2006) Seasonal (Salmon salar L). Aquaculture Food Chem. 106: 253-259. must be determined and con­ variations in chemical, physical, 277: 231-238. Paust BC, Rice AA (Eds.) (2001) trolled according to the NOM- textural, and microstructural NOM (1994) Bienes y Servicios. properties of adductor muscles Marketing and Shipping Live 031-SSA1-1993 norm (NOM, Productos de la Pesca. Crustá- Aquatic Products: Proc. 2nd Int. of Pacific lions-paw scallop ceos Fresco-Refrigerados y 1995) are the species V. chol- (Nodipecten subnodosus). Aqua- Conference and Exhibition, No- erae and Salmonella spp. In Congelados. Especificaciones vember 1999. University of culture 258: 619-632. Sanitarias. Norma Oficial Mex­ these two cases, a zero toler­ Alaska Report Nº AK-SG-01-03. Domínguez GT, Gutiérrez V (1993) icana NOM-029-SSA1-1993. Fairbanks, AK, USA. 311 pp. ance level is established for Elaboración y Evaluación de la Diario Oficial de la Federación products of this type. The sam­ Estabilidad de Salchichas Ahu- 01/02/1994. México. Riaz F, Qadri RB (1985) Quality madas de Tilapia. Thesis. Uni­ changes in lobster (Panulirus ples analyzed on day 1 and day NOM (1995) Bienes y Servicios. poliphagus) muscle during stor­ versidad de Sonora. México. Productos de la Pesca. Molus- 4 did not show the presence of 142 pp. age in ice. J. Agr. Food Chem. these bacteria; thus, the product cos Bivalvos Frescos-Refriger- 33: 117-122. Durán A, Erdemli U, Karakaya M, ados y Congelados. Especifica- does not represent a risk for Tyilmaz M (2008) Effects of ciones Sanitarias. Norma Ofi­ Ryder JM, Buisson DH, Scott DN, consumers. slaughter methods on physical, cial Mexicana NOM-031- Fletcher GC (1984) Storage of biochemical and microbiologi­ SSA1-1993. Diario Oficial de la New Zeland jack mackerel cal quality of rainbow trout Federación 03/03/1995. México. (Trachurus novaezelaniae) in Conclusions ice: Chemical, microbiological Oncorhynchus mykiss and mir­ NMX (1987) Calidad del Agua. ror carp Cyprinus carpio fil­ and sensory assessment. J. The biochemical, chemical, Determinación del Número más Food Sci. 49: 1453-1456, 1477. leted in pre-, in- or post-rigor Probable (NMP) de Coliformes and microbiological parameters periods. Fish. Sci. 74: 1146- Totales, Coliformes Fecales Ryder JM (1985) Determination of used in this study were useful 1156. (Termotolerantes) y Escherichia adenosine triphosphate and its in assessing the quality of the Hintze J (2001) NCSS and PASS, coli Presuntiva. Norma Mexi­ breakdown products in fish adductor muscle of lion’s paw Number Cruncher Statistical cana NMX-AA-042-1987. Dia­ muscle by high performance liquid chromatography. J. Agr. scallop Nodipecten subnodosus Systems. Kaysvill, Utah, USA. rio Oficial de la Federación 22/06/1987. México. Food Chem. 33: 678-680. transported and stored whole in Kawashima K, Yamanaka H (1992) Saito T, Arai K, Matsuyoshi M refrigeration. According to the Effects of storage temperatures Ocaño-Higuera VM, Pacheco-Agui­ on the post-mortem biochemical lar R, Maeda-Martínez AN (1959) A new method for esti­ analyses performed, the adduc­ changes in scallop adductor (2001) Bioquímica posmortem mating the freshness of fish. tor muscle can be considered muscle. Nipp. Suis. Gakk. 58: en pectínidos. In Maeda-Mar­ B. Jap. Soc. Sci. Fish. 24: edible until day 4 under the 2175-2180. tínez AN (Ed.) Los Moluscos 749-750. conditions of transport and Kimura M, Narita M, Nomata H, Pectínidos de Iberómerica: Sikorski ZE, Kolakowska A, Burt R Ushio H, Yamanaka H (1999). Ciencia y Acuicultura. Limusa. (1990) Postharvest biochemical storage utilized in the study. México. pp. 405-429. These results can be achieved Effects of washing methods on and microbial changes. In the rigor of scallop adductor Ocaño-Higuera VM, Maeda-Mar­ Sikorski ZE (Ed.) Seafood: Re- by maintaining the cold chain muscle. Nipp. Suis. Gakk. 65: tínez AN, Lugo-Sánchez ME, sources, Nutritional Composi- and hygienic conditions of the 103-107. Pacheco-Aguilar R (2006) Post- tion and Preservation. CRC organisms from the moment Liu S, Fan W, Zhong S, Ma C, Li P, mortem biochemical and tex­ Press. Boca Raton, FL, USA. they are harvested until their Zhou K, Zhu M (2010) Quality tural changes in the adductor pp. 55-75. commercialization. evaluation of tray-packed tilapia muscle of catarina scallop Watanabe H, Yamanaka H, Ya­ fillets stored at C based on sen­ stored at 0ºC. J. Food Biochem. makawa H (1992) Post-mortem sory, microbiological, biochem­ 30: 373-389. biochemical changes in the Acknowledgements ical and physical attributes. Afr. Ocaño-Higuera VM, Márquez-Ríos muscle of disk abalone during J. Biotechnol. 9: 692-701. E, Canizales-Dávila M, Casti­ storage. Nipp. Suiss. Gakk. 58: The authors thank Ramón Maldonado-Amparo R, Ramírez JL, llo-Yáñez FJ, Pacheco-Aguilar 2081-2088. García Arce for his technical Ávila S, Ibarra AM (2004) Trip­ R, Lugo-Sánchez ME, García- Woyewoda AD, Shaw SJ, Ke PJ, assistance during sample har­ loid lion-paw scallop (Nodipecten Orozco KD, Graciano-Verdugo Burns BG (1986) Recommended AZ (2009) Postmortem changes vest at culture site, the Consejo subnodosus Sowerby); growth, Laboratory Methods for Assess- gametogenesis, and gametic cell in cazon fish muscle stored on ment of Fish Quality. Canadian Nacional de Ciencia y Tec­ frequencies when grown at a ice. Food Chem. 116: 933-938. Technical Report of Fisheries nología (CONACYT) for fi­ high food availability site. Aqua- Ocaño-Higuera VM, Maeda-Mar­ and Aquatic Sciences Nº 1448. nancing project 59898, and culture 235: 185-205. tínez AN, Márquez-Ríos E, Nova Scotia, Canada. 143 pp.

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