Evaluation of three different microbial inhibition tests for the detection of sulphamethazine residues in the edible tissues of rabbit

The aim of the present study was to evaluate three microbial inhibition tests (MIT) based on inhibition of growth of the test organisms: (a) four plate test (FPT) containing Bacillus subtilis BGA, (b) screening test for antibiotic residues (STAR) containing Bacillus stearothermophilus var. calidolactis_ATCC 10149 and (c) the Premi®Test containing Bacillus stearothermophilus var. calidolactis. The tests were used to determine sulphamethazine (SMZ) residues in edible tissues of rabbit after oral administration up to day 15 of the withdrawal period (WP). A solvent extraction procedure was used to enhance the capability of the tests to detect SMZ residues at or below the maximum residue limit (MRL). Para-aminobenzoic acid (PABA) was employed to previously identify SMZ residues in the first stage of the residue screening. The presence of SMZ residues in the samples was confirmed and quantified by a validated HPLC method. The Premi®Test detected SMZ residues in the muscle and heart tissue up to day 9 of the WP, and in the liver, lungs and kidneys up to day 10 of the WP. The STAR detected SMZ residues in the edible organs of rabbits up to day 8 of the WP. The kidneys were positive up to day 5 of the WP, the liver until day 4 of the WP and the lungs until day 3 of the WP. No SMZ residues were detected in the muscle and heart. By using the solvent extraction procedure, SMZ residues were detected in the muscle extract up to day 10 of the WP and the muscle was positive until day 6 of the WP. No detection sensitivity was observed using the FPT. After solvent extraction, SMZ residues were detected in the muscle extract until day 8 of the WP and the muscle was positive until day 3 of the WP. No positive results were detected after the addition of PABA into/onto the agar medium. PABA at a concentration of 10 µg ml^?1 completely reversed the inhibitory activity of SMZ and enabled reliable identification of SMZ in the examined samples. Using HPLC, SMZ was detected in the muscle samples until day 10 of WP (0.02 mg kg^?1) and in the liver until day 12 of the WP (0.09 mg kg^?1). The results obtained by the HPLC method and the limit of detection (LOD) of screening tests for SMZ (FPT 0.4 µg ml^?1, STAR 0.2 µg ml^?1, Premi® Test 0.05 µg ml^?1) allowed us to state that the most suitable screening tests for the detection of SMZ residues in the edible tissues of rabbits at level corresponding to the MRL of 0.1 mg kg^?1, established for sulphonamides, are the Premi®Test and STAR in conjunction with the solvent-extraction procedure.     

Antimicrobial food packaging film based on the release of LAE from EVOH

The aim of this work was to develop antimicrobial films for active packaging applications containing the natural antimicrobial compound LAE (lauramide arginine ethyl ester) in EVOH copolymers with different mol % ethylene contents (i.e. EVOH-29 and EVOH-44). EVOH-29 and EVOH-44 films were made by casting and incorporating 0.25%, 1%, 5%, and 10% LAE in the film forming solution (w/w with respect to polymer weight). Previously, the minimum inhibitory concentration (MIC) and the minimum bactericidal concentration (MBC) of LAE against Listeria monocytogenes, Escherichia coli, and Salmonella enterica were determined by a microdilution assay. The antimicrobial activity of the resulting films was tested in vitro against these microorganisms in liquid culture media. The activity of the films was also evaluated over time. The results showed that films containing 5% and 10% LAE produced total growth inhibition and viable counts decreased with 0.25% and 1% LAE. Finally, the effectiveness of the films was tested by applying them to an infant formula milk inoculated with L. monocytogenes and S. enterica and stored for 6 days at 4°C. The application of films with LAE to infant formula milk inoculated with L. monocytogenes reduced at the end of storage period about 4 log in case of 10% LAE and with S. enterica reduced 3.74 log and 3.95 log with EVOH 29 5% and 10%, respectively, and EVOH-44 5% and 10% LAE reduced 1 log and 3.27 log, respectively, at the end of storage. The antimicrobial capacity of EVOH-29 films was greater than that of EVOH-44 films in all the cases tested. In general, the films were more effective in inhibiting the growth of L. monocytogenes than S. enterica, this inhibition being more acute at the end of the storage time.     

Updated knowledge about the presence of phenolic compounds in wine

Phenolic compounds are partly responsible for the color, astringency, and bitterness of wine, as well as for numerous physiological properties associated with wine consumption. Mass spectrometry has allowed for great progress in the identification and characterization of wine polyphenols. The aim of the present article is to summarize the numerous advances recently achieved in this field. The main type of phenolic compounds found in wine, including hydroxybenzoic and hydroxycinnamic acids, stilbenes, flavones, flavonols, flavanonols, flavanols, and anthocyanins, are firstly described. Chemical reactions and mechanisms involving phenolic compounds during winemaking are also extensively discussed, including enzymatic and chemical oxidation reactions, direct and acetaldehyde-mediated anthocyanin-tannin condensation reactions, acetaldehydemediated and glyoxylic acid-mediated tannin-tannin condensation reactions and, C-4/C-5 anthocyanin cycloaddition reactions with 4-vinylphenols, vinylflavanols and pyruvic acid, among others, leading to the formation of pyranoanthocyanins. Useful mass spectral data of well-known and novel phenolic compounds recently identified in wine, and details related to their fragmentation pathway according to different ionization techniques, are given.     

The influences of carcass weight and depot on the fatty acid composition of fats of suckling Manchego lambs

The effects of carcass weight (<5.5 kg, 5.5–6.5 kg, ⩾6.5 kg) of Manchego suckling lambs, type of fat depot of the leg (subcutaneous, intermuscular and intramuscular) and muscle (m. longissimus dorsi (LD) or m. quadriceps femoris (QF)) on the fatty acid composition were studied. Carcass fatness increased with increasing carcass weight. However, few differences in fatty acid proportion were observed between the three carcass weight groups. The lightest carcasses had lower proportions of myristic (C14:0) and palmitoleic (C16:1) acid and higher proportions of stearic (C18:0) acid, desirable fatty acids (DFA) (C18:0 + total unsaturated fatty acid (TUFA)) and a greater nutritive value ((C18:0 + C18:1)/C16:0). The fatty acid composition of subcutaneous and intermuscular fat depots of the leg were similar, whereas the intramuscular fat depot was different from both of these, since it had a lower proportion of saturated fatty acids (SFA), a greater proportion of TUFA, higher PUFA/SFA and n − 6/n − 3 ratios, and greater DFA and nutritive values. The intramuscular fat of the LD was more saturated than QF, higher SFA, and had lower value of PUFA/SFA ratio and DFA. These results indicated that the intramuscular fat quality of the leg was better than subcutaneous and intermuscular fat depots and also that intramuscular fat displayed better attributes of quality (lower SFA, higher TUFA, and greater PUFA/SFA ratio and DFA as well) than intramuscular fat of the LD from the consumer health standpoint. Despite carcass weight affected carcass fatness, it did not seem to influence the proportions of saturated or polyunsaturated fatty acids. For this reason, carcass weight probably had little effect on the organoleptic characteristics of the meat.     

Characterization and Technological Properties of Lactic Acid Bacteria Isolated from Traditional Argentinean Goat’s Milk Products

A total of 286 lactic acid bacteria isolated from goat’s dairy products in northwest of Argentina were characterized. Lactobacilli (38%) and cocci (62%) were identified according to morphological, physiological, and relevant technological properties. L. plantarum (14%) and Enterococcus (34%) were the predominant species. S. thermophilus, Pediococcus and L. plantarum were the highest acid producers. Eight strains of L. fermentum produced bacteriocins or metabolites similar to bacteriocins. The API-ZYM test was applied to 39 isolates. Eight strains were selected from their both technological properties and enzymatic activities for use as starter or adjunct culture in the manufacture of artisanal goat cheeses.     

Microencapsulation of β‐carotene using native pinhão starch,modified pinhão starch and gelatin by freeze‐drying

Beta‐carotene was microencapsulated by freeze‐drying using native pinhão starch, hydrolysed pinhão starch 6 dextrose equivalent (DE), hydrolysed pinhão starch 12 DE and the mixture of these materials with gelatin as coating material. The purpose of this research was to produce and characterize these microcapsules. The capsules’ efficiency, surface content, moisture, morphology, solubility, particle size and glass transition temperature were analysed. The hydrolysed pinhão starch 12 DE showed the highest total β‐carotene content and the lowest surface β‐carotene content, unlike the native starch. Using scanning electron microscopy, it was observed that all microcapsules presented undefined shapes. The samples with gelatin had wider particle size distribution, higher diameters, lower solubility and higher glass transition temperature when compared with other the samples. Results obtained suggest that the modified pinhão starch can be considered as potential wall material for encapsulation of β‐carotene.     

Changes in the volatile composition of red wines during aging in oak barrels due to microoxygenation treatment applied before malolactic fermentation

Microoxygenation is a wine-making technique consisting in the addition of small and controlled amounts of oxygen. This study has examined the effect of this technique on the volatile composition of two red single variety wines during two successive vintages. The microoxygenation treatment was applied at the end of alcoholic fermentation and before beginning malolactic fermentation. Once the microoxygenation treatment had finished, wines were aged in new American oak barrels for 12 months. The results obtained showed that the microoxygenation treatment did not cause significant changes in the varietal and fermentation volatile compounds, however microoxygenation slowed down the extraction of some of the volatile compounds extracted from wood. A varietal and vintage effect was also observed for some of the compounds studied.     

Modifications in membrane fatty acid composition of Salmonella typhimurium in response to growth conditions and their effect on heat resistance

The effects of growth temperature (in the range 10-45 degrees C) and acidification up to pH 4.5 of the culture medium (Brain Heart Infusion, BHI) with different organic acids (acetic, citric and lactic) and hydrochloric acid on membrane fatty acid composition and heat resistance of Salmonella typhimurium CECT 443 were studied. The heat resistance was maximal in cells grown at 45 degrees C (cells grown in non-acidified BHI showed a D58-value of 0.90 min) and decreased with decreasing growth temperature up to 10 degrees C (D58-value of 0.09 min). The growth of cells in acidified media caused an increase in their heat resistance. In general, acid adapted cells showed D-values of between 1.5 and 2 times higher than the corresponding for non-acid adapted control cells. This cross-protection response, which has important implications in food processing, was not dependent on the pH value and the acid used to acidify the growth medium. A membrane adaptation corresponding to an increase in the unsaturated to saturated fatty acids ratio (UFA/SFA) and membrane fluidity was observed at low growth temperature. Moreover, the acidification of the growth medium caused a decrease in UFA/SFA ratio and in the C18:1 relative concentration, and an increase in cyclopropane fatty acids (CFA) content mainly due to the increase in cyc19 relative concentration. Thus, acid adapted cells showed CFA levels 1.5 times higher than non-acid adapted control cells. A significant proportion of unsaturated fatty acids were converted to their cyclopropane derivatives during acid adaptation. These changes in membrane fatty acid composition result in cells with decreased membrane fluidity. A clear relation between membrane fatty acid composition and heat resistance was observed. In general, D-values were maximum for cells with low UFA/SFA ratio, and, consequently, with low membrane fluidity. Moreover, CFA formation played a major role in protecting acid adapted cells from heat inactivation. However, changes observed in membrane fatty acid composition are not enough to explain the great thermotolerance of cells grown at 45 degrees C. Thus, other mechanisms, such as the synthesis of Heat Shock Proteins, could be responsible for this increase in the bacterial heat resistance.     

Genomic merit for reproductive traits. I: Estrous characteristics and fertility in Holstein heifers

Genetic selection of dairy cattle in the United States has included reproductive traits (daughter pregnancy rate, DPR; heifer conception rate, HCR), which is believed to have partly contributed to halting the decline in reproductive performance. The objectives of the current study were to evaluate the association among genomic merit for DPR (GDPR) and HCR (GHCR) with estrous characteristics measured by an automated device. Holstein heifers (n = 1,005) were genotyped at 2 mo of age and were classified into quartiles (Q1 = lowest, Q4 = highest) according to the GDPR and GHCR values of the study population. At 10 to 11 mo of age, heifers were fitted with a collar that recorded activity and rumination and determined the occurrence of estrus according to changes in activity and rumination compared with the individual's baseline values. Estrous characteristics of spontaneous estruses (SPE) and PGF_2α-synchronized estruses (PGSE) were recorded. Heifers had their estrous cycle synchronized with PGF_2α and following detection of estrus received either artificial insemination or embryo transfer according to the herd's genetic selection program. Heifers in Q2 (17.7 ± 0.3 h) of GHCR tended to have longer SPE than heifers in Q4 (16.7 ± 0.3 h). The interaction between GDPR and GHCR was associated with the likelihood of activity peak (0 = no estrus, 100 = maximum activity) ≥80 at SPE because, among heifers in Q3 and Q4 of GHCR, those in Q1 of GDPR were less likely to have an activity peak ≥80. Heifers in Q1 and Q2 of GDPR had reduced hazard of estrus within 7 d of the first PGF_2α treatment compared with heifers in Q4 of GDPR. Heifers in Q1 (16.1 ± 0.4 h) of GDPR had shorter PGSE than heifers in Q2 (17.6 ± 0.4 h) and Q4 (17.4 ± 0.4 h) and tended to have shorter PGSE than heifers in Q3 (17.4 ± 0.4 h). Rumination nadir on the day of PGSE was greater for heifers in Q1 (?30.1 ± 0.9 min/d) of GDPR compared with heifers in Q4 (?33.7 ± 0.9 min/d). Among heifers receiving only artificial insemination, those in Q1 of GHCR (adjusted hazard ratio = 0.65; 95% confidence interval = 0.48–0.88) became pregnant at a slower rate than heifers in Q4. Genomic merit for HCR was negatively associated with SPE but tended to be positively associated with hazard of pregnancy, whereas GDPR was positively associated with PGSE and hazard of estrus. Selection of dairy cattle for DPR and HCR may improve reproductive performance through different pathways, namely estrous characteristics and pregnancy establishment.