Production of Interesting Peptide Fractions by Enzymatic Hydrolysis of Tuna Dark Muscle By-Product Using Alcalase

ABSTRACT

A protein hydrolysate was prepared from proteins of tuna dark muscle by-product. The hydrolysis conditions (time, temperature, pH, and enzyme concentration) using Alcalase was optimized by response surface methodology (RSM). The regression coefficient close to 1.0, observed during experimental and validation runs, indicated the validity of the model. The hydrolysate produced under the optimum conditions determined by RSM has a low rate of peptide fraction of molecular weight of 4–1 kDa. Meanwhile, the results obtained by hydrolysis under optimal conditions determined by a complementary study (temperature 55°C, time 60 min, 1% enzyme concentration, and pH 8.5) show that the hydrolysate produced has a height rate of the peptide fraction of molecular weight of 4–1 kDa. The amino acid composition of the protein hydrolysate prepared proved to have the potential for application as an ingredient in balanced fish diets and as a source of nitrogen in microbial growth media.     

Occupancy pattern of a long-horned beetle in a variegated forest landscape: linkages between tree quality and forest cover across spatial scales

Context

Interactions between landscape-scale processes and fine-grained habitat heterogeneity are usually invoked to explain species occupancy in fragmented landscapes. In variegated landscapes, however, organisms face continuous variation in micro-habitat features, which makes necessary to consider ecologically meaningful estimates of habitat quality at different spatial scales.

Objectives

We evaluated the spatial scales at which forest cover and tree quality make the greatest contribution to the occupancy of the long-horned beetle Microplophorus magellanicus (Coleoptera: Cerambycidae) in a variegated forest landscape.

Methods

We used averaged data of tree quality (as derived from remote sensing estimates of the decay stage of single trees) and spatially independent pheromone-baited traps to model the occurrence probability as a function of multiple cross-scale combinations between forest cover and tree quality (with scales ranging between 50 and 400 m).

Results

Model support and performance increased monotonically with the increasing scale at which tree quality was measured. Forest cover was not significant, and did not exhibit scale-specific effects on the occurrence probability of M. magellanicus. The interactive effect between tree quality and forest cover was stronger than the independent (additive) effects of tree quality and particularly forest cover. Significant interactions included tree quality measured at spatial scales ≥200 m, but cross-scale interactions occurred only in four of the seven best-supported models.

Conclusions

M. magellanicus respond to the high-quality trees available in the landscape rather than to the amount of forest per se. Conservation of viable metapopulations of M. magellanicus should consider the quality of trees at spatial scales >200 m.

     

Performance and carcass characteristics of lambs fed diets with increasing levels of <Emphasis Type="Italic">Mimosa tenuiflora</Emphasis> (Willd.) hay replacing Buffel grass hay

This study evaluated the performance and carcass characteristics of lambs fed diets with increasing levels of Mimosa tenuiflora (Willd.) hay replacing Buffel grass (Cenchrus ciliaris). Twenty-eight Santa Inês male lambs with an average body weight (BW) of 20.3 ± 1.49 kg(mean ± SD) were allocated in individual stalls and distributed in a completely random design with four treatments (0, 20, 40, and 60 g/100 g total DM M. tenuiflora hay replacing Buffel grass hay in diet) with seven replications. M. tenuiflora hay at the level of 20% dry matter (DM) total replacing Buffel grass hay increased final weight (P = 0.006), total weight gain (P < 0.001), average daily weight gain (ADWG; P < 0.001), DM intake (P < 0.001), and feed efficiency (P < 0.001). Intake of crude protein, NDF_ap, ADF_ap, ash, ether extract, total and non-fibrous carbohydrates, and total digestible nutrients presented a positive quadratic effect with M. tenuiflora hay replacing Buffel grass hay and 40 g/100 g total DM level presented greater intake. There were positive quadratic effects by M. tenuiflora hay inclusion at 20 g/100 g total DM level on slaughtering weight (P = 0.005), hot carcass weight (P = 0.002), cold carcass weight (P = 0.002), empty body weight (P = 0.001), hot carcass yield (P = 0.002), cold carcass yield (P = 0.003), and increase linear on biological yield (P = 0.003). There was no influence on cooling weight loss (P = 0.284). M. tenuiflora hay may be included in lamb diets at amounts up to 20 g/100 g total DM substitution of Buffel grass hay because increase in the nutrients intake, growth performance, and carcass characteristics.     

Identification of pathogenic Leptospira strains in tissues of a premature foal by use of polymerase chain reaction analysis.

Studies were carried out to determine the cause of death in a prematurely born Thoroughbred foal that died 24 hours after birth. Necropsy revealed gross lesions suggestive of septicemia. A commercial Leptospira polymerase chain reaction (PCR) assay designed to specifically amplify the hemolysis-associated protein 1 (hap1) gene present only in pathogenic Leptospira strains detected the presence of Leptospira DNA in various tissues of the foal. Histologic examination of lung, liver, kidney, and myocardium revealed numerous spirochetes in Warthin-Starry-stained tissue sections. Results of PCR analysis and histologic examination suggested a leptospiral infection in the newborn foal. At the moment of death, the infection coexisted with a streptococcal-associated aspiration bronchopneumonia and postpartum septicemia. These findings indicate that the PCR assay based on the amplification of the hap1 gene represents a useful tool for specific detection of pathogenic leptospira in field samples taken from horses.     

Alteration of body composition in rats: effect of organic chromium and L-carnitine

A growth trial was carried out to test the effect of organic, trivalent chromium and L-carnitine on the body composition of growing rats. At the same time, an evaluation of different measurement methods (weight of epididymal fat pad, adipocyte morphometry, total body electrical conductivity) was performed. Outbred Wistar rats of 30 days of age were fed diets of different (0, 10 and 20%) protein level. The diets were supplemented with 4 mg/kg Cr as chromium nicotinate, and 100 mg/kg L-carnitine. The experimental feeding lasted 15 days, after a 5-day-long adjustment period. It was found that Cr addition increased feed intake. Both treatments caused changes in body composition, increasing fat and protein deposition. Organic chromium had no effect at either protein level, while L-carnitine improved the protein retention only at an optimum (20%) protein supply. No statistically significant correlation was found between total body electrical conductivity (TOBEC) and body composition, which could be attributed to the great individual differences. A close correlation was found among total body fat percentage, weight of epididymal fat pad and the adipocyte surface. The data suggest that there is an interaction between dietary protein supply and the effect of repartitioning agents.     

Pseudorabies virus infection in mink: a host-specific pathogenesis

Pseudorabies virus (PRV) is an alphaherpesvirus that causes a neurological disease in many wild and domestic animals. The neuropathology elicited by PRV is quite consistent regardless of the host with the only exception of mink, in which it is characterized by a vasculopathy rather than by an encephalitis. In this study, we aimed to investigate the underlying pathogenic mechanism(s) of PRV infection in mink by using immunohistochemistry and laser capture microdissection (LCM) on material from naturally and experimentally infected animals. The inflammatory reaction induced by PRV was minimal or absent not only in the nervous system, where we identified a low number of macrophages and a few T lymphocytes, but also in the primary replication site, the oropharyngeal mucosa; however, the number of PRV-infected cells detected by immunohistochemistry was extremely high both in the peripheral mucosa and in the nervous tissue. On the other hand, the vascular pathology included parenchymal hemorrhages of various degrees and, in specific cortical areas of the brain, fibrinoid degeneration of the capillary walls. Detection of viral antigens by immunohistochemistry revealed infection of endothelial cells of capillaries situated both in the oropharyngeal mucosa and in the brain stem; the presence of PRV DNA in vessels was further demonstrated by PCR performed on LCM samples of brain capillaries. These results can be interpreted as supporting the idea that the different pathology of the disease in mink may be the consequence of an increased endotheliotropism of PRV in this species. Infection of the vessel wall may then lead to vascular pathology and impairment in endothelial cell function, resulting in a weak immune response to infection.     

Evaluation of endotoxaemia in the prognosis and treatment of scouring merino lambs

This study looked at measurement of endotoxaemia as a tool in determining prognosis and probable response to treatment in scouring lambs. One hundred eighty-three lambs in the first 15-20 days of life, from eight Merino sheep farms located in the region of La Serena, south-west Spain, were used in this experiment. Scouring and normal/control lambs were selected following a clinical examination, the scouring group was further divided into subgroups, specifically those that did or did not survive 72 h following treatment. At the time of the clinical examination, faecal and blood samples were taken. Faecal culture and commercial faecal antigen tests for detection of enteropathogens in faeces and serum endotoxin measurement using chromogenic lymulus amoebocyte lysate (LAL) were carried out. Scouring lambs received 0.07 mg/kg liveweight halofuginone once a day for 3 days, a single oral dose of 0.20 mg/kg liveweight of spectinomycin and oral rehydration fluid. The pathogens isolated were Cryptosporidium spp. and Escherichia coli. The case fatality rate was 51% in the scouring lambs. Postmortem findings were consistent with enterotoxigenic E. coli infection. The concentration of endotoxin was 0.18 +/- 0.12 ng/ml in the control group, 0.35 +/- 0.17 ng/ml in the surviving lambs and 0.46 +/- 0.14 ng/ml in the non-surviving lambs. Significant differences between groups were found. Case fatality rate of the scouring lambs with endotoxaemia below 0.30 ng/ml was 0%, while it was 100% above 0.50 ng/ml. These results may be utilized as a prognostic indicator in lambs affected by E. coli and Cryptosporidium that will help aid in decision-making as to whether to treat a lamb or not based on its chances of survival.     

Effect of different fat sources on in vitro degradation of nutrients and certain blood parameters in sheep

This study was designed to determine the effects of calcium salt of palm oil fatty acids (CS), hydroxyethylsoyamide (HESA), butylsoyamide (BSA) and soybean oil (SO) on degradation of crude protein and fibre in vitro, and on the blood plasma lipid parameters in vivo. Five mature wethers (body weight 75 kg) were fed five diets in a 5 x 5 Latin square experiment. The control diet consisted of 50% meadow hay and 50% concentrate with no added fat. The control diet was supplemented with CS, HESA, BSA, or SO. Fat was added at 3.5% of dietary dry matter (DM). The final ether extract content of the ration was near 6%. Each period lasted 20 days. Fat supplements, except HESA, consistently decreased the in vitro DM disappearance of soybean meal as compared to control. In contrast to the effect of other treatments, crude protein degradation was greatest in the test tubes with inocula obtained from sheep fed diet with HESA. Fat supplements equally inhibited the DM and fibre breakdown of alfalfa pellet. CS and HESA seemed to be less detrimental to in vitro fermentation of neutral detergent fibre (NDF) than BSA and SO. All fat supplements increased blood plasma triglyceride, cholesterol and total lipid content. Plasma concentration of cholesterol and total lipid was highest with SO. The inclusion of CS in the diet increased 16:0, while all fat supplements increased plasma 18:0 and decreased 16:1 and 18:1 fatty acid content. Plasma 18:2n-6 was not changed by feeding CS and SO. However, compared to the control diet, 18:2n-6 increased with 12 and 41% in plasma fatty acids when sheep were fed HESA and BSA, respectively. The results showed that plasma concentration of linoleic acid was enhanced more when the amide was synthesised from butylamine than when from ethanolamine.     

Evaluation of the on-line electrical conductivity of milk in mastitic dairy cows

Mastitis is a persistent, inflammatory reaction of the udder tissue, which entails a decline in potassium, and is also responsible for a higher somatic cell count (SCC) and electrical conductivity (EC) of milk. The measurement of EC is an indirect, rapid method to detect subclinical mastitis from milk. The aim of this study was to analyse the EC of milk throughout the lactation, around the day of mastitis detection, and also to estimate its heritability based on data from a large-scale dairy farm. Shortly after calving the EC value generally decreases; however, it was discovered that from the thirteenth week onwards, substantial differences arise between the mastitic and healthy groups of cows. The authors observed a significant (P < 0.001) increase in EC before the detection of clinical mastitis. This higher value (around 11 mS) persisted for 4 days, then it gradually returned to the normal level. The EC of milk from daughters sired by different bulls responded differently in case of mastitis. The heritability of EC seems to be high (h2 = 0.56). Therefore, the EC trait can be a beneficial indicator in detecting mastitis and should be considered in sire selection.