Bimol Chandra Roy

Effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle of male broilers

1. The effects of nutritional level on muscle development, histochemical properties of myofibre and collagen architecture in the pectoralis muscle were evaluated using male broilers of Red Cornish × New Hampshire stock, reared on diets of high nutritional value for up to 80 d (H80d) and low nutritional value for up to 80 d (L80d, same age as H80d) or 95 d (L95d, same body weight as H80d). 2. The total live weight and the weight of pectoralis muscle were lower in L80d than in both H80d and L95d. The muscle weight as a percentage of live weight was 8·7% in L80d, 10·7% in H80d and 11·5% in L95d. 3. Pectoralis muscle was composed only of type IIB myofibres and showed no differences in myofibre type composition among the chicken groups. The largest diameter of type IIB myofibres was observed in L95d, followed by H80d and the smallest in L80d. 4. The total amount of intramuscular collagen did not differ among the chicken groups (1·92 to 1·99 mg/g). Types I and III collagens were immunohistochemically detected in both the perimysia and endomysia. The thin perimysia around the primary myofibre fascicles showed larger width in H80d than L80d and L95d, and also the thick perimysia around the secondary fascicles in H80d than L80d. 5. The collagen structure of the perimysium was most developed in H80d, followed by L95d and on the least in L80d. The development of perimysial collagen fibres could be enhanced by a rapid growth rate of the muscle induced by high nutritional level and depressed by a slow growth rate with low nutritional foods. 6. The endomysial collagen architecture was observed as a felt-like tissue of the fibril bundles with many slits. The thinnest endomysial wall was observed in L80d, followed by H80d and the thickest in L95d. 7. From these results, it was indicated that foods of high nutritional value could enhance growth of the pectoralis muscle of broilers, and this is accompanied by hypertrophy of the type IIB myofibres and development of the perimysial collagen architecture.

Structural and ultrastructural changes of full-cycle cultured Pacific bluefin tuna (Thunnus orientalis) muscle slices during chilled storage.

BACKGROUND This study examined the structural and ultrastructural changes of dorsal and ventral muscle tissues of full-cycle cultured Pacific bluefin tuna (PBT), Thunnus orientalis Temminck & Schlegel 1844, cut into slices simulating sashimi and placed in chilled storage for varying periods. Structural and ultrastructural changes were determined in order to understand the physical texture by breaking strength measurement. RESULTS Progressive deterioration of myofibril structure was observed during chilled storage (4 °C) of PBT muscle slices over 5 days post mortem. Muscle degradation included detachment between myofibres, detachment of the plasmalemma, disruption of mitochondria, loss of Z-line density and alignment, cementation of myofibrils, loss of the hexagonal arrangement of thick versus thin myofilaments and migration of subsarcolemmal nuclei to intermyofibrillar spaces. CONCLUSION Loss of myofibre-myofibre adhesion, detachment of the plasmalemma and disruption of other components did not lower the breaking strength of PBT muscle. This provides evidence that the muscle breaking strength of PBT is not only associated with the detachment of myofibres or detachment of the plasmalemma. Other factors that produce cement-like substances, such as cementation of the myofibrillar components and degradation of the sarcoplasmic reticulum, may also increase breaking strength.

Collagen content and architecture of the pectoralis muscle in male chicks and broilers reared under various nutritional conditions

Varying chicken growth rates were induced with different nutritional regimes, and the collagen content and architecture of M. pectoralis (PT) were compared among 21-day-old chicks and broilers at 80 or 95 days of age. The percentage of muscle weight to live weight was higher in rapid growing chicks (8.4%) than slow growing chicks (6.3%). The 80-day-old broilers engaged in compensatory growth after the early slow growth period producing PT muscle at 11% of live weight. The 80- and 95-day-old chicks with restricted late growth after an early rapid growth period showed PT weight at 8% and 9% of live weight, respectively. Collagen content of the PT muscle markedly decreased from the chicks to the broilers. The collagen concentration was higher in the late-growth restricted broilers (1.67-1.88 mg/g) than the compensatory growth broilers (1.01-1.10 mg/g). Collagen concentration did not differ between the rapid and slow growing chicks (2.72 and 2.94 mg/g). Scanning electron micrographs showed thick and thin perimysia, and honeycomb endomysia. In the perimysia, a stack layer of collagen platelets and a reticular layer of collagen fiber cords were distinguished and collagen baskets of adipocytes were observed. The perimysial collagen fibers became thicker during growth of the chicks to broilers. However, in the late-growth restricted broilers, the perimysial collagen fibers seemed to have retarded development compared with the compensatory growth birds. The PT muscle of chickens develops optimally when body growth is enhanced. The PT muscle of the compensatory growth broilers had improved collagen architecture regardless of the marked decrease in collagen content.

Histochemical properties and collagen architecture of M. iliotibialis lateralis and M. puboischiofemoralis in male broilers with different growth rates induced by feeding at different planes of nutrition

1. The histochemical properties and the collagen content and architecture of the iliotibialis lateralis (ITL) and puboischiofemoralis (PIF) muscles were assessed in Red Cornish × New Hampshire cockerels reared on a high nutrient plane for 80 d (H80d), or a low nutrient plane for 80 d (L80d) or 95 d (L95d). 2. Final live weights were 3410 g in H80d, 2810 g in L80d and 3467 g in L95d. Both ITL and PIF muscle weights were lowest in L80d and did not differ between H80d and L95d. 3. ITL muscle was composed of fast-twitch myofibres such as IIA (high reduced nicotinamide adenine dinucleotide dehydrogenase, NADH-DH activity), IIB (low NADH-DH activity) and IIC (intermediate NADH-DH activity). The high percentage of type IIB myofibres in H80d (76·6%) and L95d (76·2%) birds were reflected in low percentages of type IIC myofibres (12·2%) in H80d birds and type IIA myofibres (8·2%) in L95d birds. Percentages of IIA, IIB and IIC myofibres in L80d cockerels were 12·4, 69·8 and 17·6%, respectively. 4. The myofibres in PIF muscle were divided into two basic types, I and IIA, and a transitional form (I-tr) from IIA to I. In the caudal region, all myofibres in H80d and L95d cockerels were type I but in L80d cockerels 15% of myofibres were categorised as type I-tr. In the cranial region, the great majority (52 to 63%) of myofibres were type IIA. Type I myofibres occurred at a higher percentage in H80d (30·5%) than L95d (21·8%) and type I-tr in L95d (15·7%) than H80d (7·3%) and L80d (11·5%). 5. The total amount of collagen was higher in ITL than PIF muscle in every bird group. In both muscles the highest collagen content was in L95d cockerels but the content did not differ between H80d and L80d birds. The thickness of thick and thin perimysia increased with muscle size. The circular collagen fibre in the thick perimysium was larger in ITL (6·1 to 7·0 µm) than PIF (3·7 to 3·8 µm) muscle but did not differ among the bird groups. 6. From these results, it was concluded that feeding on a high nutritional plane promotes growth of the thigh muscles, with accompanying enlargement of the perimysial thickness, no increase in collagen content and various changes of histochemical properties.

Modification of mature non-reducible collagen cross-link concentrations in bovine m. gluteus medius and semitendinosus with steer age at slaughter, breed cross and growth promotants.

Increased meat toughness with animal age has been attributed to mature trivalent collagen cross-link formation. Intramuscular trivalent collagen cross-link content may be decreased by reducing animal age at slaughter and/or inducing muscle re-modeling with growth promotants. This hypothesis was tested in m. gluteus medius (GM) and m. semitendinosus (ST) from 112 beef steers finished at either 12 to 13 (rapid growth) or 18 to 20 (slow growth) months of age. Hereford-Aberdeen Angus (HAA) or Charolais-Red Angus (CRA) steers were randomly assigned to receive implants (IMP), ractopamine (RAC), both IMP and RAC, or none (control). RAC decreased pyridinoline (mol/mol collagen) and IMP increased Ehrlich chromogen (EC) (mol/mol collagen) in the GM. In the ST, RAC increased EC (mol/mol collagen) but decreased EC (nmol/g raw muscle) in slow growing CRA steers. Also, IMP increased ST pyridinoline (nmol/g raw muscle) of slow-growing HAA steers. Results indicated alteration of perimysium collagen cross-links content in muscle in response to growth promotants.

Proximate and Fatty Acid Compositions in Different Flesh Cuts of Cultured, Cultured Fasted, and Wild Pacific Bluefin Tuna (Thunnus orientalis)

This study was carried out to determine the differences in proximate and fatty acid (FA) compositions in different flesh cuts of the most valuable cephalic portion of cultured (full-cycle), cultured fasted (full-cycle), and wild Pacific bluefin tuna (PBT; Thunnus orientalis). Cultured fasted PBTs were fasted for 6 days to ensure empty stomachs during harvesting and to determine the influence of fasting on the lipid and FA composition in different flesh cuts. Significantly higher lipid contents were observed (p < 0.05) in the dorsal akami (lean part) and the ventral o-toro (very fatty part) flesh cuts in cultured and cultured fasted PBTs than in wild ones. However, monounsaturated fatty acid (MUFA) contents in all flesh cuts were significantly higher in cultured and cultured fasted PBTs than in wild PBT. Wild PBTs contained significantly higher (p < 0.05) polyunsaturated fatty acid (PUFA) contents than other cultured PBTs in all flesh cuts except for the dorsal akami and wakaremi (fatty part near dorsal fin) flesh parts, and contained higher n-3 FA content than other PBTs—especially docosahexaenoic acid (DHA, C22:6n-3)—in the lipid fraction from the dorsal chu-toro (medium fatty part), ventral akami, ventral chu-toro, o-toro, and in dark flesh cuts. The flesh quality of cultured or cultured fasted PBTs has the advantage of having a higher total amount of essential FAs than that of wild PBT.

Understanding the quality of typical and atypical dark cutting beef from heifers and steers

Dark cutting beef with pH <6.0 may have glucidic potential approaching that of normal beef but has increased toughness that may also be influenced by cattle sex and carcass characteristics. Beef longissimus thoracis (LT) from normal (Canada AA; n=24), and typical (TB4; pH>5.9, n=20) and atypical (AB4; pH<5.9, n=20) dark cutting carcasses were analyzed to investigate relationships between beef quality, pH, glucidic potential and carcass characteristics. Results indicated that reduced lactate and glucidic potential were accompanied by increased pH and reduced L*, a* and b* values. Mean glucidic potential was lowest (P<0.0001) for TB4 whereas glucidic potentials for heifers and steers AB4 LT were sufficient to attain normal pH, substantiating the existence of atypical dark cutting. Warner-Bratzler shear force of AB4 remained higher (P<0.05) than that of Canada AA LT confirming AB4 as the toughest beef irrespective of sex and carcass characteristics.

Pretreatment with formic acid enhances the production of small peptides from highly cross-linked collagen of spent hens

Small collagen peptides are associated with various benefits, such as bone and skin health. However, preparation of small collagen peptides from terrestrial vertebrate origins remains a challenge. Here, we show that pretreatment with formic acid enhanced the production of small collagen peptides from spent hen skin. After pretreatment with formic acid, the percentage of small peptides below 2 kDa increased to 48.92% and 43.34% from 33.79% and 36.32% for heat-soluble collagen (HSC) and pepsin-soluble collagen (PSC), respectively. Pretreatment with formic acid degraded telopeptides and released the cross-links (pyrrole and pyridinoline), which made hen collagen more susceptible to papain hydrolysis. LC-MS/MS results revealed that none of the peptides identified from HSC-FA (formic acid)-Papain and PSC-FA-Papain were derived from cross-linked telopeptides. These results demonstrated that formic acid assisted the hydrolysis of highly cross-linked collagen of spent hens, and it might also be used to produce small collagen peptides from other aged, vertebrate collagens.

Removing Cross-Linked Telopeptides Enhances the Production of Low-Molecular-Weight Collagen Peptides from Spent Hens

The low-molecular-weight (LMW) peptides derived from collagen have shown a potential for various nutritional and pharmaceutical applications. However, production of LMW peptides from vertebrate collagen remains a challenge. Herein, we report a new method to produce LMW collagen peptides using pepsin pretreatment that removed cross-linked telopeptides in collagen molecules. After the pretreatment, the proportion of LMW collagen peptides (<1.4 kDa) that were obtained from pepsin-soluble collagen increased to 32.59% compared to heat-soluble collagen peptides (16.10%). Fourier transform infrared spectroscopy results indicated that telopeptide cleavage retained the triple-helical conformation of collagen. Liquid chromatography-tandem mass spectrometry analysis suggested that Gly-X-Y (X is often proline, while Y is either hydroxyproline or hydroxylysine) repeats were not the main factors that hindered the enzymatic hydrolysis of collagen molecules. However, cross-link quantification demonstrated that trivalent cross-links that included pyridinolines and pyrroles were the primary obstacles to producing small peptides from collagen of spent hens. This study demonstrated for the first time that removing cross-linked telopeptides could enhance the production of LMW peptides from spent hen collagen, which is also of interest to manufacturers who produce LMW collagen peptides from other vertebrate animals, such as bovids and porcids.

Effects of Brief Summer or Winter Fasting on the Muscle Quality of Cultured Pacific Bluefin Tuna (Thunnus orientalis)

The effects of fasting on the quality of the dorsal and ventral ordinary muscles from cultured Pacific bluefin tuna (Thunnus orientalis) during chilled storage were investigated. Tuna were subjected to fasting for 2 days in the summer or 6 days in the winter prior to harvesting. The breaking strength of the dorsal ordinary muscle sampled in the summer increased until 24 h and then decreased. There were no significant differences in the lipid and glycogen content of the ordinary muscle after 9 h of storage between the controls and either fasting group. The pH of the ordinary muscle subjected to summer and winter fasting was higher than in the controls after 24–48 h of storage. However, the relationship between the pH and glycogen content was unclear. The metmyoglobin content during chilled storage was lower in the ordinary muscles from either fasting group than in the controls. In conclusion, fasting for 6 days in the winter improved the color stability of the ordinary muscle without a decline in its lipid content.