Activities of mu- and m-calpain and of calpastatin were measured at four different times during postmortem storage (0, 1, 3, and 10 d) in three muscles from either callipyge or noncallipyge (normal) sheep. The weights of two muscles, the biceps femoris and the longissimus, are greater in the callipyge phenotype, whereas the weight of the infraspinatus is not affected. The activity of m-calpain was greater (P < 0.05) in the biceps femoris and longissimus from callipyge than in those from normal sheep, but it was the same in the infraspinatus in the two phenotypes. The extractable activity of m-calpain did not change (biceps femoris and infraspinatus) or decreased slightly (longissimus) during postmortem storage. Extractable activity of mu-calpain decreased to zero or nearly zero after 10 d postmortem in all muscles from both groups of sheep. The rate of decrease in mu-calpain activity was the same in muscles from the callipyge and normal sheep. At all time points during postmortem storage, calpastatin activity was greater (P < 0.05) in the biceps femoris and longissimus from the callipyge than from the normal sheep, but it was the same in the infraspinatus from callipyge and normal sheep. Calpastatin activity decreased (P < 0.05) in all three muscles from both phenotypes during postmortem storage; the rate of this decrease in the callipyge biceps femoris and longissimus and in the infraspinatus from both the callipyge and normal sheep was slow, especially after the first 24 h postmortem, whereas calpastatin activity in the biceps femoris and longissimus from the normal sheep decreased rapidly. During postmortem storage, the 125-kDa calpastatin polypeptide was degraded, but the 80-kDa subunit of mu-calpain was cleaved only to 76- and 78-kDa polypeptides even though extractable mu-calpain activity declined nearly to zero. Approximately 50 to 60% of total mu-calpain became associated with the nonextractable pellet after 1 d postmortem. The myofibril fragmentation index for the biceps femoris and longissimus from normal sheep increased significantly during postmortem storage. The fragmentation index for the infraspinatus from the callipyge and normal sheep increased to an intermediate extent, whereas the index for the biceps femoris and longissimus from the callipyge did not change during 10-d postmortem storage. The results suggest that postmortem tenderization is related to the rate of calpastatin degradation in postmortem muscle and that calpastatin inhibition of the calpains in postmortem muscle is modulated in some as yet unknown manner.
The knowledge of consumer perception of meat tenderness and taste is essential to forecast a Brazilian quality value-based beef market. This study aimed to verify perception of tender (WBSF ≤ 4.1 kg) from tough (≥ 4.8 kg) strip loin steak or uncharacteristic (calcium-treated/Ca-IM) and normal (noncalcium/NO-Ca) meat taste by consumers according to gender, age, education and income levels. Steaks were previously classified by shear force measurements as tender or tough. Each consumer was served a paired sample of one tender and of one tough steak, which were either Ca-IM or NO-Ca treated before tenderness classification. Three hundred and eight consumers answered a nine-point intensity (tenderness) and hedonic (taste) scales evaluation questionnaire. Among consumers, 82.2% indicated beef as first choice meat products, 75.3% had beef at least four times a week; 39.3% considered taste as the most important meat attribute and 30.2% considered tenderness; 75.8% were males; 73.6% were 21 to 55 years old; 56.7% had college education; 76.6% had monthly income higher than US $ 435,00. Tender steaks were scored highest (P < 0.01), independently of gender, age and income. However, elderly consumers gave higher scores to tender steaks in comparison to middle age consumers (P < 0.05). In the lower education level, scores given to tender and tough meat did not differ. The higher income level responders assigned lower tenderness scores within tender or tough meat (P = 0.10). Differences in taste were perceived by both genders, and by consumers in every income and education level. Males gave higher scores (dislike less) within Ca-IM steaks. Consumers in the lower education level scored taste higher (like most) within untreated samples. The elderly people could not differentiate taste between the Ca-IM and NO-Ca steaks. These are the first indications that Brazilian consumers perceive tender from tough or uncharacteristic taste of beef, but palatability is evaluated differentially depending on gender, age, education and income level.
Properties of the calpain bound to myofibrils in longissimus muscle from callipyge or noncallipyge sheep were examined after 0, 1, 3, and 10 d of postmortem storage at 4 degrees C. Western analysis has shown that most of this calpain is mu-calpain, although the sensitivity of the antibodies used in the earlier studies could not eliminate the possibility that up to 10% of the calpain was m-calpain. The calpain is bound tightly, and very little is removed by washing with the detergent Triton X-100; hence, it is not bound to phospholipids in the myofibril. Over 25% of total mu-calpain was bound to myofibrils from at-death muscle, and this increased to approximately 40% after 1 d postmortem. The amount of myofibril-bound mu-calpain increased only slightly between 1 and 10 d of postmortem storage. The percentage of autolyzed mu-calpain increases with time postmortem until after 10 d postmortem, when all myofibril-bound mu-calpain is autolyzed. The specific activity of the myofibril-bound calpain is very low and is only 6 to 13% as high as the specific activity of extractable mu-calpain from the same muscle. It is unclear whether this low specific activity is the result of unavailability of the active site of the myofibril-bound calpain to exogenous substrate. The myofibril-bound calpain degrades desmin, nebulin, titin, and troponin T in the myofibrils, and also releases undegraded alpha-actinin and undergoes additional autolysis when incubated with Ca2+; all these activities occurred slowly considering the amount of myofibril-bound calpain. Activity of the myofibril-bound calpain was partly (58 to 67%) inhibited by the calpain inhibitors, E-64 and iodoacetate; was more effectively inhibited by a broader-based protease inhibitor, leupeptin (84 to 89%); and was poorly inhibited (43 to 45%) by calpastatin. Release of undegraded alpha-actinin and autolysis are properties specific to the calpains, and it is unclear whether some of the myofibril-bound proteolytic activity originates from proteases other than the calpains or whether the active site of myofibril-bound calpain is shielded from the inhibitors. Activities and properties of the myofibril-bound calpain were identical in longissimus muscle from callipyge and normal sheep, although previous studies had indicated that the "normal" longissimus was much more tender than the callipyge longissimus. Hence, it seems unlikely that the myofibril-bound calpain has a significant role in postmortem tenderization of ovine longissimus.
This study unveils histological features of the intestinal tract of juvenile striped catfish Pseudoplatystoma fasciatum (Linnaeus, 1776) in three size classes (weight, standard length): I – 36.84 ± 10.19 g, 14.52 ± 1.54 cm; II – 59.03 ± 11.47 g, 17.17 ± 1.06 cm; III – 89.72 ± 18.70 g, 20.79 ± 1.55 cm, respectively. Histological organization of the juvenile speckled catfish intestine bears features common to the carnivorous fish, but the organ presents some convolutions that indicate a certain degree of dietary flexibility, a surprising trend, common only to omnivorous Siluriforms. The architecture of the mucosa of the speckled catfish intestine indicates that the species concentrates digestion and absorption of nutrients in the medium intestine, a common feature among carnivorous Teleosts.
Histological responses of the intestine are key for evaluating nutritional value of feed ingredients, since the organ is not only the chief site of feed digestion and nutrient absorption but also plays an important immunological function. Histomorphological alterations were evaluated in the intestine of juvenile striped catfish, Pseudoplatystoma fasciatum, fed diets containing 0 (control), 10 or 20% inclusion of lyophilized bovine colostrum (LBC), as source of protein or bioactive peptides, for either 30 or 60 days. Fish fed 20LBC presented at 60d a distinct pattern of macrophages and, some of them, higher number of vacuoles in rectum mucosa. The thickness of the muscle layer (TML) in fish fed diets with LBC was higher in the first portion of medium intestine than fish fed 0LBC. All fish presented significant increase of TML in the second portion of medium intestine along feeding period, but fish fed 20LBC had smaller values of TML than those of fish fed 0 and 10LBC which might be related to the higher intestinal coefficient found for this group. The TML of rectum was higher just for fish fed 10LBC. Dietary LBC altered morphometrical features of juvenile striped catfish intestine and possibly induced inflammatory reaction in the rectal mucosa, as a function of level of inclusion, feeding period and segment of intestine analyzed. Key words: gut absorption, histology, mammal protein, lyophilized bovine colostrum, striped catfish Histomorfologia intestinal de Pseudoplatystoma fasciatum alimentado com colostro bovino como fonte de proteína e peptídeos bioativos RESUMO: Respostas histológicas do intestino são fundamentais para avaliar o valor nutritivo de ingredientes alimentares, uma vez que o órgão não é só o principal local de digestão e absorção dos nutrientes, mas também exerce uma importante função imunológica. Alterações histomorfológicas foram avaliadas no intestino de juvenis de cachara, Pseudoplatystoma fasciatum, alimentado com dietas contendo 0 (controle), 10 e 20% de inclusão de colostro bovino liofilizado (CBL) como fonte de proteína e peptídeos bioativos, aos 30 e 60 dias. Aos 60 dias, peixes alimentados com 20CBL apresentaram macrófagos de aspecto distinto e alguns, uma grande concentração de vacúolos na mucosa retal. A espessura da camada muscular (ECM) em peixes alimentados com CBL foi maior na primeira porção do intestino médio em relação àqueles alimentados com 0CBL. Ao longo do período experimental, todos os peixes mostraram aumento significativo na ECM na segunda porção do intestino médio, entretanto peixes do grupo 20CBL tiveram menores valores para a ECM do que os demais, o que pode estar relacionado com o maior coeficiente intestinal encontrado para este grupo. A ECM no reto foi maior apenas para os peixes alimentados com 10CBL. A inclusão de CBL na dieta do cachara alterou características morfométricas do intestino e, possivelmente, induziu uma reação inflamatória na mucosa retal em função do nível de inclusão, período de alimentação e porção do intestino analisada. Palavras-chave:...
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