Milk-Clotting Properties and Primary Proteolysis of a Lyophilised Extract from Artichoke Flowers (Cynara scolymus L.).

Research background: A few studies have investigated Cynara scolymus enzymes as a substitute for calf rennet in cheese making. They used aqueous extracts prepared by maceration of plant material and stored by freezing. However, it was indicated that lyophilisation is a better alternative to preserve the coagulant properties of plant extracts over a longer period of time, as it is a more controllable, stable and hygienic alternative with a better shelf life that is easier to transport, store and standardise. Experimental approach: We obtained a lyophilised extract of mature artichoke flowers, named CS, which was characterised for its milk-clotting properties at different pH and temperatures. In addition, the potential yield, whey composition and the primary hydrolysis profile of caseins by urea-polyacrylamide gel electrophoresis (PAGE) of mini curds prepared with different doses of coagulant were determined. Results and conclusions: The lyophilised extract was able to hydrolyse casein and showed stable proteolytic activity at pH=6.4 and 37 °C for 50 min, which decreased when the process temperature was increased to 41 and 45 °C and was lost at 70 °C. On the other hand, milk-clotting activity increased significantly (p<0.001) when the temperature increased from 37 to 45 °C and the pH of the milk decreased from 6.8 to 5.8. Potential yield ​​between 10 and 17 % was obtained for the mini curds prepared with the lyophilised artichoke extract, and the casein degradation pattern obtained by urea-PAGE was similar to that of the commercial coagulant. Novelty and scientific contribution: On a laboratory scale, our work has shown that the lyophilised artichoke extract has sufficient proteolytic and coagulant activity to be used as a milk coagulant, i.e. plant rennet, in cheese making as an alternative to animal rennet. As this extract is lyophilised, it has the advantage of being a better alternative in terms of preservation and shelf-life. It offers an innovative way to diversify cheese products and appeal to consumers with different dietary preferences and needs.

Prediction of tenderness in bovine longissimus thoracis et lumborum muscles using Raman spectroscopy.

OBJECTIVE: This study was conducted to evaluate Raman spectroscopy technique as a noninvasive tool to predict meat quality traits on Braford longissimus thoracis et lumborum muscle. METHODS: Thirty samples of muscle from Braford steers were analyzed by classical meat quality techniques and by Raman spectroscopy with 785 nm laser excitation. Water holding capacity (WHC), intramuscular fat content (IMF), cooking loss (CL), and texture profile analysis recording hardness, cohesiveness, and chewiness were determined, along with fiber diameter and sarcomere length by scanning electron microscopy. Warner-Bratzler shear force (WBSF) analysis was used to differentiate tender and tough meat groups. RESULTS: Higher values of cohesiveness and CL, together with lower values of WHC, IMF, and shorter sarcomere were obtained for tender meat samples than for the tougher ones. Raman spectra analysis allows tender and tough sample differentiation. The correlation between the quality attributes predicted by Raman and the physical measurements resulted in values of R2 = 0.69 for hardness and 0,58 for WBSF. Pearson's correlation coefficient of hardness (r = 0.84) and WBSF (r = 0.79) parameters with the phenylalanine Raman signal at 1,003 cm-1, suggests that the content of this amino acid could explain the differences between samples. CONCLUSION: Raman spectroscopy with 785 nm laser excitation is a suitable and accurate technique to identify beef with different quality attributes.

Is beef quality affected by the inclusion of distiller grains in cattle diets?

This study aimed to analyze the effect of including increasing levels of distiller grains (DG) in beef cattle feeding diets on meat quality from an integral approach. To this end, we analyzed the meat from 36 yearling steers fed with four dietary treatments: 0DG (control corn-based diet), 15DG, 30DG, and 45DG (containing 15%, 30%, or 45% of DG on DM basis). Crude protein, ash, and fat contents of diets increased with DG level. The fatty acid profile of DG diets was reflected in the fatty acid profile of plasma samples. Feeding diets with DG had no effect on the biochemical parameters analyzed in plasma. In addition, it did not influence the water holding capacity, the muscle or fat color at 72 h post mortem or at retail display, or the contents of SFAs, MUFAs, and PUFAs in beef. The DG diets led to lower values of cooking loss. Meat from 15DG and 30DG showed greater percentage of troponin C and fragments of 30 to 27 kDa than meat from 45DG. Also, meat from 30DG showed the lowest values of Warner-Bratzler shear force. Hence, including up to 30% corn DG in beef cattle feeding diets had positive effects on meat quality.

Dietary inclusion of ruminally protected linseed oil as a means to mitigate heat and slaughter-induced stress in feedlot cattle.

There is evidence of a relationship between increased energy intake and the development of metabolic inflammation and insulin resistance (IR), and between the aforementioned metabolic state and impaired tolerance to heat stress. Based on the anti-inflammatory properties and mitigating effects on IR and stress of n-3 polyunsaturated fatty acids (n-3 PUFA), an experiment was performed to evaluate the effect of n-3 PUFA supplementation to feedlot-finished steers during summer on animal performance, physiological and biochemical variables associated with glucose metabolism, heat and preslaughter-induced stress, and meat quality. A total of 48 Angus steers (388 ± 2 kg) were fed one of three corn-based finishing diets containing (dry matter basis) 0% added oil (CON; negative control), or 1.90% of sunflower oil-calcium salt (SUN; positive control), or 1.92% of linseed oil-calcium salt (LIN). There was a trend (P = 0.08) for greater dry matter intake (DMI) and greater (P = 0.02) average daily gain (ADG) in LIN-fed animals compared with the average between those that received the CON or SUN diets, whereas no differences (P ≥ 0.34) were observed between the latter. No other performance, physiological, or carcass variables were affected (P ≥ 0.12) by treatment. Blood glucose and insulin were similar (P ≥ 0.14), though the homeostatic model assessment (HOMA) which gauges IR tended (P = 0.06) to be reduced for LIN-fed animals compared with the average between those that received the CON or SUN diets. Blood insulin and HOMA increased linearly (P ≤ 0.01) with days on feed. An interaction between the study phase (feeding period or slaughter) and treatment was observed (P ≤ 0.05) for glucose and cortisol. While the magnitude of glucose increase (P < 0.01) from the end of the feeding period to slaughter was greater for CON- and SUN-fed animals compared with LIN-fed ones, cortisol increased (P < 0.05) only in animals that received CON or SUN diets. Meat quality attributes were not affected (P ≥ 0.16) by treatment. The concentration of n-3 PUFA was greater (P < 0.01) and n-6:n-3 ratio was lesser (P < 0.01) in meat from LIN-fed animals compared with that resulting from the average between the animals that received the negative (CON) or positive (SUN) control diets. Results suggest that n-3 PUFA supplementation mitigated metabolic alterations associated with IR and preslaughter-related stress. It may have also improved tolerance to heat, resulting in greater DMI and ADG of steers fed a high-energy diet during summer. Results also indicate that glucose metabolism and heat stress tolerance worsen with time when feeding concentrate-based diets.

Polyunsaturated fatty acids (n-3 PUFA) of diets have anti-inflammatory properties and mitigating effects on insulin resistance (IR) and stress in steers. This study evaluated the effect of n-3 PUFA supplementation to feedlot-finished steers during summer on animal performance, physiological and biochemical response, and meat quality. We found that n-3 PUFA supplementation mitigated metabolic alterations associated with IR and slaughter-related stress. It may have also improved tolerance to heat, resulting in greater dry matter intake, and average daily gain of steers fed a high-energy diet during summer.

Effect of Aging and Retail Display Conditions on the Color and Oxidant/Antioxidant Status of Beef from Steers Finished with DG-Supplemented Diets.

The aim of this work was to study the effect of finishing diets including distiller grains (DG) on color and oxidative stability of beef after being exposed to aerobic retail display conditions, with or without previous aging. For this purpose, beef samples from animals fed with finishing diets including 0%, 15%, 30%, and 45% DG (on a dry matter basis), which had been exposed to aerobic retail display conditions, with or without previous aging under vacuum packaging, were evaluated. The content of γ-tocopherol, ß-carotene, and lutein in diet samples increased with the level of DG. Beef evaluated at 72 h post-mortem showed greater content of γ-tocopherol and retinol as the DG level increased. Meat color was not affected by DG inclusion, but color parameters decreased with retail conditions. Meat from animals fed with DG showed the lowest values of thiobarbituric acid reactive substances (TBARS), independently of the retail display conditions. However, all samples were below the threshold associated with rancid aromas and above the a* value related to meat color acceptance. Thus, feeding diets including up to 45% of DG improved the antioxidant status of meat, preserving the color, and delaying lipid oxidation in meat samples under the display conditions evaluated.

Feeding strategies and ageing time alter calpain system proteins activities and meat quality of Braford steers.

OBJECTIVE: The aim of this study was to evaluate the effect of ageing and feeding strategies on the calpain protease system and meat quality traits in Braford steers. METHODS: Thirty Braford steers were employed; 15 animals were supplemented with corn silage during finishing and 15 were kept only on pasture. Meat quality traits and calpain system protein activity were evaluated in longissimus thoracis et lumborum (LTL) steaks aged for 2, 7, 14, and 21 days. RESULTS: Aged meat showed higher pH and calcium content, while Warner Bratzler shear force (WBSF) decreased to day 21. No interaction between ageing and diet was seen for quality traits. Steers finished with corn silage showed higher values of water holding capacity, WBSF and free calcium, and lower values of pH and cooking loss. Calpain and calpastatin activities decreased with ageing. Finishing steers on pasture produced higher values of calpains and lower values of calpastatin activities. The higher values of calpain 1 activity were observed in muscles aged 2 days from pasture finished animals, and the lower activity of the inhibitor in the 21 days aged samples of the same group. CONCLUSION: These results suggest a diet by ageing interaction in calpains and calpastatin and this interaction impact in Warner Bratzler Shear Force in Braford LTL muscle.

Color and Marbling as Predictors of Meat Quality Perception of Argentinian Consumers.

In Argentina, color and intramuscular fat are the main attributes of raw beef quality; however, it is necessary to clarify how consumers use them, in order to establish different marketing strategies. Consumer preferences are affected by multiple factors. Thus, the objective of the present study was to identify the Argentinian consumer's choice behavior regarding beef color and fat content. An online survey was carried out in Argentina. It inquired about socio-demographic characteristics, purchase and consumption habits and beliefs, showing pictures related to color, marbling and the amount of fat. Choice behavior was evaluated by asking why consumers chose a particular picture out of the ones shown. Several Kruskal-Wallis tests evaluated the different hypotheses. Three different decision trees using the CHAID analysis method were created. Multifactorial analysis was carried out for clustering consumers. Regarding consumer beliefs, 90% of the respondents agreed with the sentence, "The two main characteristics defining beef quality at purchase time are meat color and marbling". Socio-demographic characteristics affected purchase habits and beliefs; they also affected perceptions about meat color and marbling. It was possible to build three consumer groups for future marketing strategies: "hedonic" focused on a pleasing sensory experience, "appearance" prioritized the visual aspects, and the "health-conscious" consumers were interested in their healthy nutrition.

Combination of organic acids and low-dose gamma irradiation as antimicrobial treatment to inactivate Shiga toxin-producing Escherichia coli inoculated in beef trimmings: Lack of benefits in relation to single treatments.

The aim of this study was to assess the efficacy of lactic acid (LA), caprylic acid (CA), high- (HDI) and low- (LDI) dose gamma irradiation and LDI combined with LA or CA on the inactivation of a pool of Shiga toxin-producing Escherichia coli (STEC) strains inoculated on beef trimmings. The three most efficacious treatments were selected to study their effect on meat quality parameters and sensory attributes. The inoculum included five native STEC serogroups (O26, O103, O111, O145 and O157). The treatments applied were 0.5% LA, 0.04% CA, 0.5 kGy LDI, 2 kGy HDI, LDI+LA and LDI+CA. Beef trimmings were divided into two groups; one was inoculated with high (7 log CFU/g) and the other with low (1 log CFU/g) level of inoculum. Efficacy was assessed by estimating log reduction and reduction of stx- and eae-positive samples after enrichment, respectively. Results showed that treatments with organic acids alone were not effective in reducing STEC populations. For high inoculum samples, the most effective treatment was HDI followed by LDI+LA and LDI alone or combined with CA. For low inoculum samples, the most effective treatment was HDI followed by LDI alone or combined with organic acids. Concerning meat quality parameters and sensory attributes, irradiation treatments (LDI and HDI) caused minimal changes, while LDI+LA modified them significantly compared with the control. Therefore, based on our results, no benefits were observed after combining organic acids with gamma irradiation.

Feeding strategies alter gene expression of the calpain system and meat quality in the longissimus muscle of Braford steers.

OBJECTIVE: The aim of the present study was to determine the effect of supplementing pasture-finished steers with corn silage on the expression level of the calpain system proteins and beef tenderization. METHODS: Thirty Braford steers grazing on summer pasture were used for the study. For 120 days fifteen animals were supplemented with corn silage at 1% of body weight per head per day (Suppl) whereas the remaining 15 steers only received pasture (Contr). Carcass and meat traits were evaluated and compared between groups. Gene expression and activities of proteases (calpain 1 and calpain 2) and inhibitor (calpastatin) were measured using real-time polymerase chain reaction and casein zymography. RESULTS: Carcass and meat traits were significantly different between feeding systems. Supplemented steers showed higher hot carcass weight (p<0.01), fat content (p = 0.02), and Warner-Bratzler shear force (p = 0.03). Furthermore, the control group showed higher protease:inhibitor ratios, at mRNA (p = 0.01) and protein levels (p<0.10). Warner-Bratzler shear force and mRNA calpains:calpastatin ratio were associated in both feeding systems (p<0.01). CONCLUSION: Based on the results obtained in the study, beef tenderness differences among finishing strategies could be modulated through differential expression of the calpain system proteins.

A Contribution of Beef to Human Health: A Review of the Role of the Animal Production Systems.

Meat and meat products constitute important source of protein, fat, and several functional compounds. Although beef consumption may implicate possible negative impacts on human health, its consumption can also contribute to human health. Quality traits of beef, as well as its nutritional properties, depend on animal genetics, feeding, livestock practices, and post mortem procedures. Available data show that emerging beef production systems are able to improve both, quality and nutritional traits of beef in a sustainable way. In this context, Argentina's actions are aimed at maximising beef beneficial effects and minimising its negative impact on human health, in a way of contributing to global food security.

Galacturonosyltransferase 4 silencing alters pectin composition and carbon partitioning in tomato.

Pectin is a main component of the plant cell wall and is the most complex family of polysaccharides in nature. Its composition is essential for the normal growth and morphology pattern, as demonstrated by pectin-defective mutant phenotypes. Besides this basic role in plant physiology, in tomato, pectin structure contributes to very important quality traits such as fruit firmness. Sixty-seven different enzymatic activities have been suggested to be required for pectin biosynthesis, but only a few genes have been identified and studied so far. This study characterized the tomato galacturonosyltransferase (GAUT) family and performed a detailed functional study of the GAUT4 gene. The tomato genome harbours all genes orthologous to those described previously in Arabidopsis thaliana, and a transcriptional profile revealed that the GAUT4 gene was expressed at higher levels in developing organs. GAUT4-silenced tomato plants exhibited an increment in vegetative biomass associated with palisade parenchyma enlargement. Silenced fruits showed an altered pectin composition and accumulated less starch along with a reduced amount of pectin, which coincided with an increase in firmness. Moreover, the harvest index was dramatically reduced as a consequence of the reduction in the fruit weight and number. Altogether, these results suggest that, beyond its role in pectin biosynthesis, GAUT4 interferes with carbon metabolism, partitioning, and allocation. Hence, this cell-wall-related gene seems to be key in determining plant growth and fruit production in tomato.

Effect of aging on the characteristics of meat from water buffalo grown in the Delta del Paraná region of Argentina.

Fifteen crossbreed water buffalos were selected from a farm in Delta del Paraná, Entre Ríos Province, Argentina. Animals were castrated males 20-24 months old reaching final live weights of 400-420kg. They were predominantly of Mediterranean and Murrah breeds and were feed in naturally grown pastures. Tenderness and chewiness increased with postmortem aging (p<0.05). Aging did not affect flavour and odour scores, even though certain off-flavours and off-odours were reported. Changes in colour with aging were similar to those seen in beef.