Broccoli processing wastes as a source of peroxidase

A peroxidase isozyme (BP) was purified to homogeneity from broccoli stems ( Brassica oleraceae var. maraton) discarded from industrial processing wastes. BP specific activity was 1216 ABTS [2,2'-azinobis(3-ethylbenzthiazoline-6-sulfonic acid)] units/mg, representing 466-fold that of crude extract. BP is a monomeric glycoprotein containing 16% carbohydrates, with a molecular mass of 49 kDa and an isoelectric point close to 4.2. From kinetic data it showed a two-substrate ping-pong mechanism, and the catalytic efficiency measured as the rate-limiting step of free BP regeneration was 3.4 x 10(6) M(-1) s(-1). The ABTS K m value was 0.2 mM, which was about 20 times lower than that reported for acidic commercial horseradish peroxidase (HRP). Assessment of BP secondary structure showed 30% helical character, similar to HRP and cytochrome c peroxidase. BP lost only 25% activity after 10 min of heating at 55 degrees C and pH 6; it was stable in the pH range from 4 to 9 and showed an optimum pH of 4.6 using ABTS as substrate. BP was active on substrates normally involved in lignin biosynthesis, such as caffeic and ferulic acids, and also displayed good catechol oxidation activity in the presence of hydrogen peroxide. Reverse micellar extraction was successfully used as potential large-scale prepurification of broccoli peroxidase, achieving a purification factor of 7, with 60% activity yield. Stems from the broccoli processing industry have a high potential as an alternative for peroxidase purification.     

Multivariate analysis for the evaluation of fiber, sugars, and organic acids in commercial presentations of table olives

Table olives constitute an important part of the Mediterranean diet and the diet of many non-olive-producing countries. The aim of this work was to determine the fiber, sugar, and organic acid contents in Spanish commercial presentations of table olives and characterize them by means of a multivariate analysis. The selection of variables was carried out on the basis of a canonical analysis and their classification, according to processing styles and cultivars, through a linear discriminant analysis. Values of dietary fiber in table olives ranged from 2 to 5 g/100 g edible portion (e.p.). Some stuffing materials (almond, hot red pepper, and hazelnut) or the addition of capers produced a significant increase in the total dietary fiber in green olives. Glucose, fructose, and mannitol were usually found in the ranges of 0-55, 0-70, and 0-107 mg/100 g e.p., respectively. Succinic acid was detected only in green and directly brined olives (0-40 mg/100 g e.p.), while lactic and acetic acids were used within the ranges of 0-681 and 5-492.8 mg/100 g e.p., respectively. A multivariate analysis showed that fiber, mannitol, and succinic, lactic, and acetic acids can be used to discriminate between processing styles (95.5% correct assignations) and cultivars (61.20%). Current data can also be used in the evaluation of the dietary value of table olives.     

Supplemental feeds containing coffee pulp for rearing Tilapia in Central AmericaAlimento suplementario conteniendo pulpa de café para la crianza de Tilapia en Centro America

A supplementary feed containing 30% coffee pulp was evaluated for use in the culture of Tilapia aurea in El Salvador, Central America. A comparison of the coffee pulp feed with a feed containing all of the same ingredients except coffee pulp was made with T. aurea raised in 1.0-m³ cages suspended in a fertilized earthen pond. Survival was high in all treatments and there was no significant difference in average weight gain between groups of fish receiving the two experimental feeds. Production trials were conducted in 100-m² fish pens and in 0.05-ha earthen ponds. Pen-raised fish receiving coffee pulp feed grew faster throughout the experiment, and total production was approximately twice that in control treatments. Highest production in pens was 1.25 kg/m² per year. Results of production pond trials using T. aurea at 9 000/ha and stocked with the piscivorous Cichlasoma managuense yielded an estimated 3 392 kg/ha per year in fed treatments and 2 049 kg/ha per year in controls. Low feed conversion (1.92) and low feed cost resulted in an increase in net annual earnings of $251.00/ha.     

Metabolic effects of changes in the dietary protein: carbohydrate ratio in eel (Angilla anguilla) and trout (Oncorhynchus mykiss)

The metabolic consequences of the isoenergetic replacement of dietaryprotein by carbohydrates (CHO) were compared in studies of European eel andrainbow trout. Diets with 45/20, 37/30, 29/40 and 21/50 percentprotein/carbohydrate were assessed during a 12-week experiment. The CHO sourcewas pre-gelatinized corn starch. Fish of initial average weight 45gwere fed to satiation twice daily. Weight-gain and feed-efficiency data weregenerally better in trout than in eel, presumably because trout digestibilitycoefficients for protein and energy were higher than in eel. In both species,regardless of physiological differences in digestion and absorption, feedintakeand nutrient digestibility were lower when dietary CHO was higher, but theinter-species differences decreased with increasing dietary CHO.Pyruvate kinase activity (PK) in trout was not affected by dietary CHO content,while blood-glucose and liver-glycogen levels significantly rose withincreasingdietary CHO. In contrast, blood-glucose levels appeared to be regulated ineels,perhaps by an accelerated glycolysis rate, revealed by changes in PK.Gluconeogenic activity was inhibited in trout fed a diet containing20–30%CHO, while in eels this activity was not inhibited by dietary replacement ofprotein by carbohydrate. Liver glucose 6-phosphate dehydrogenase activity(G6PDH) was inhibited in trout fed increasing dietary CHO, whereas in eelsG6PDHoperated at a high rate regardless of dietary composition. These results mayindicate that eels have a better capacity for metabolising high-CHO/low-proteindiets than do trout.     

Expression of a glycoprotein hormone receptor gene in the ovary of the bullseye puffer (Sphoeroides annulatus)

A fragment of a glycoprotein hormone receptor (GpHR) gene was isolated from the ovary of the bullseye puffer. This cDNA fragment showed similarity to the thyrotropin (TSH) and gonadotropin (LH and FSH) receptors. GpHR mRNA expression in the ovary gradually increased during oocyte maturation.     

Marketing of meat sheep with intensive finishing in southern state of Mexico

The objective of the present study was to quantify the marketing margins of cold sheep carcass and barbacoa meat in the southern state of Mexico. Data were obtained from the production chain links (production, industrialization, and marketing) where the marketing margins were calculated along with the benefit/cost (B/C) ratio. In the absolute marketing margin of the final consumer price per kilogram of carcass meat, the producer obtained US$2.7/kg (47 %) of the utilities, while the intermediaries obtained US$3.1/kg (53 %). Considering the final cooked product in barbacoa (typical dish), the margin was US$6.3/kg (29 %) for the producer and US$15.2/kg (71 %) for the intermediaries. The B/C ratio was 1.0, 1.1, 2, and 1.3 for the producer, stocker, butcher, and barbacoa seller, respectively. It is concluded that the best marketing channel for the producer was the producer-stocker-processor (butcher and barbacoa seller). The highest marketing margin was for the intermediaries followed by the producer. The order of importance of the B/C kilogram ratio of meat was for the butcher first, then barbacoa seller, and lastly stocker and producer.     

Detection of anti-Leishmania infantum antibodies in sylvatic lagomorphs from an epidemic area of Madrid using the indirect immunofluorescence antibody test

An outbreak of human leishmaniosis was confirmed in the southwest of the province of Madrid, Spain, between July 2009 and December 2012. Incidence of Leishmania infection in dogs was unchanged in this period, prompting a search for alternative sylvatic infection reservoirs. We evaluated exposure to Leishmania in serum samples from animals in the area with an indirect immunofluorescence test (IFAT). Using promastigotes from six culture passages and a 1/25 threshold titer, we found anti-Leishmania infantum seroreactivity in 9.3% of cats (4 of 43), 45.7% of rabbits (16/35) and 74.1% of hares (63/85). Use of promastigotes from >10 in vitro passages resulted in a notably IFAT lower titer, suggesting antigenic changes during extended culture. Postmortem inspection of seropositive animals showed no clinical signs of infection. The results clearly suggest that asymptomatic hares were the main reservoir in the outbreak, and corroborate IFAT as a sensitive serological surveillance method to detect such cryptic Leishmania infections.