Depletion study of enrofloxacin and its metabolite ciprofloxacin in edible tissues and feathers of white leghorn hens by liquid chromatography coupled with tandem mass spectrometry

To ensure delivery of safe foods to consumers, withdrawal times for drugs must be respected according to the maximum residual limits established by regulatory agencies. Because of availability and price, feather meal is currently incorporated into animal feed as a protein source for farm species. Few data are available on residual drugs in feathers from treated animals. A depletion study was performed with laying hens treated intramuscularly with 5% enrofloxacin (Enromic) at 10 mg/kg body weight over 3 days. Thirty-three birds were treated and slaughtered at different times between 6 and 216 h after treatment; and samples of muscle plus skin, liver, kidney, and feathers were collected. High-performance liquid chromatography coupled with a tandem mass spectrometry method was validated before sample analysis to determine the decision limit, detection capability, recovery, and precision. Liver was the edible tissue with the slowest drug depletion. A withdrawal time of 6 days was calculated based on European Union maximum residual limits (100 microg/kg). A withdrawal time of 9 days was calculated based on Japan maximum residual limits (10 microg/kg). Enrofloxacin plus ciprofloxacin concentrations in feathers remained high through all sampling periods. Thus, feathers from treated animals should not be fed to food-producing animals.     

Assessment of safety of enterococci isolated throughout traditional Terrincho cheesemaking: virulence factors and antibiotic susceptibility

Enterococci account for an important fraction of the adventitious microflora of traditional cheeses manufactured in Mediterranean countries from small ruminants' raw milk and play an important role in the development of suitable organoleptic characteristics of the final product. It has been suggested that animals used for food or animals that supply edible products are a reservoir of antibiotic-resistant enterococci. The main purpose of this research effort was thus to identify, to the species level, a total of 73 enterococci with high tolerance to acidic pH and bile salts (as prevailing environmental conditions in the first portion of the gastrointestinal tract), which were previously isolated from the milk feedstock to the final product of Terrincho cheesemaking, and to determine their profiles of antibiotic susceptibility, coupled with the occurrence of specific virulence factors (especially in those that might eventually be claimed to exhibit suitable probiotic and technological performances). Isolates, identified by both API 20 STREP and PCR methods, were found to belong to the following Enterococcus species: E. casseliflavus, E. durans, E. faecalis, E. faecium, and E. gallinarum. Susceptibility of those isolates was observed to most antibiotics tested, whereas none harbored aminoglycoside resistance genes. PCR screenings for cytolysin genes (cylL(L), cylL(s), cylM, cylB, and cylA), surface adhesin genes (efaA(fs), efaA(fm), and esp), the aggregation protein gene (agg), and the extracellular metalloendopeptidase gene (gelE) were performed. All isolates proved negative for cylL(L), cylM, cylB, and agg genes. Both E. faecalis strains were positive for the cell wall-associated protein Esp and the cell wall adhesin efaA(fs), whereas the cell wall adhesin efaA(fm) was detected in 11 of the 12 E. faecium strains. Only one strain possessed the cylL(s) determinant, and another possessed the cylA gene. Incidence of virulence determinants was thus very low; hence, the enterococcal adventitious microflora tested is essentially safe.     

Antimicrobial susceptibility of Listeria monocytogenes isolated from food and clinical cases in Navarra, Spain

The susceptibility of 440 Listeria monocytogenes strains isolated from food (n=401) and clinical cases (n=39) between 1995 and 2005 was determined by standard agar dilution and E-test methods. Antimicrobial drugs currently used in veterinary and human therapy were tested, and they included penicillin G, ampicillin, cephalothin, gentamicin, chloramphenicol, tetracycline, doxycycline, trimethoprim, erythromycin, and clindamycin. The sensitivity of strains was established using Clinical and Laboratory Standards Institute (formerly NCCLS) breakpoints and MIC50 (the MIC for 50% of the strains) to MIC90 values. In general, isolates were susceptible to the majority of the antimicrobials tested, including beta-lactamics and aminoglycosides, which are normally used in the treatment of listeriosis. Resistance to tetracycline and doxycycline was found in five strains isolated from fresh trout belonging to the same fish farm. Molecular analysis by restriction endonuclease analysis showed a similar profile, suggesting the persistence of a strain well adapted to the presence of tetracycline in the environment of a fish farm, which is frequently used in aquaculture in order to prevent infections of fish.     

Survey of temperature and consumption patterns of fresh-cut leafy green salads: risk factors for listeriosis

Increasing demand for fresh-cut or ready-to-eat fruits and vegetables, developed to meet the consumer need for quick and convenient products, has prompted extensive research into their microbiological quality, safety, processing, and packaging. The microbial ecology of Listeria monocytogenes is recognized as a major safety concern for fresh-cut produce. A survey was performed to collect information on consumption patterns of fresh-cut leafy green salads and the temperature of domestic refrigerators. Salad consumption was low-moderate: 24.3% of respondents never purchased fresh-cut leafy green salads; of those who reported buying these products, 7.41% did so more than twice a week, 17.28% once or twice a week, 29.63% once or twice a month, and 45.68% occasionally. Saving time and convenience were the advantages most widely reported by consumers. A total of 9.9% of respondents did not always respect the "use-by" date of fresh-cut salads, a negative practice that could contribute to the risk of listeriosis. Temperatures reported in domestic refrigerators were compatible with the growth of L. monocytogenes on ready-to-eat salads. Variations in average temperature followed a normal distribution, N(6.62, 2.56), while the variability of temperature variance was described by a gamma distribution, G(2.00, 1.00). As expected, when a time of day-temperature profile was plotted over a 24-h period, changes corresponding to the transition between day and night were observed. Knowledge of consumption patterns and consumer hygiene practices is essential, first in assessing the risk of listeriosis (risk assessment) and second in taking measures to manage that risk (risk management).     

Effect of the inoculation level of Lactobacillus acidophilus in probiotic cheese on the physicochemical features and sensory performance compared with commercial cheeses

The complex metabolism of probiotic bacteria requires several technological options to guarantee the functionally of probiotic dairy foods during the shelf life. This research aimed to evaluate the effect of the supplementation of increasing amounts of Lactobacillus acidophilus (0, 0.4, or 0.8 g/L of milk) on the physicochemical parameters and sensory acceptance of Minas fresh cheese. In addition, the sensory acceptance of probiotic cheeses was assessed using a consumer test and compared with commercial cheeses (conventional and probiotic). High counts (9.11 to 9.42 log cfu/g) of L. acidophilus were observed throughout the shelf life, which contributed to the maintenance of its probiotic status and resulted in lower pH values and greater production of organic acids. The probiotic cheeses presented lower scores for appearance, aroma, and texture compared with conventional cheeses. Internal preference mapping explained almost 60% of the total variation of the data and showed a large number of consumers concentrated near the conventional cheeses, demonstrating greater preference for these samples. The findings indicated that some negative sensory effects could occur when high level of supplementation with L. acidophilus is used in probiotic cheese processing.     

Precipitation of low molecular weight phenolic compounds of grape seeds cv. Carménère (<Emphasis Type=Italic>Vitis vinifera</Emphasis> L.) by whole saliva

Low molecular weight phenolic compounds (LMWP) contribute to astringency and bitterness, two important sensory attributes. This work aimed to study the interactions between human saliva and a group of LMWP from a grape seed extract, namely, monomers, dimers and trimers of flavan-3-ol plus gallic acid. LMWP in the sediment that was produced by mixing whole saliva with the grape seed extract were identified by high-performance liquid chromatography analysis (HPLC–DAD). Two assays to produce LMWP-salivary protein sediments were carried out. An indirect assay consisted of in vitro mixing aliquots of saliva with a constant volume of the grape seed extract followed by centrifugation and sediment analysis. In a direct assay, aliquots of the grape seed extract were mixed in-mouth with saliva, returned to a vessel and centrifuged. In each assay, polyphenol composition of the sediments varied according to the tannin/saliva vol/vol ratio although in both cases monomeric LMWP were the most abundant polyphenol components of the sediments. The study also suggested the need for a strict control of the in vitro experimental conditions used to mimic the in vivo conditions in which tannin–protein interactions do occur and produce astringency perception.     

Evaluation of aeration and substrate concentration on the production of carotenoids by <Emphasis Type="Italic">Sporidiobolus salmonicolor</Emphasis> (CBS 2636) in bioreactor

This study aimed at optimising the cultivation conditions for the production of carotenoids by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor. The maximum content of total carotenoids in the full factorial design 22 was 3131.3 μg/L in synthetic medium with 80 g/L of glucose, 15 g/L of peptone, 5 g/L of malt extract, aeration of 1.5 vvm, agitation of 180 rpm, initial pH of 4.0 at 25 °C. In the kinetic study, we could observe that the bioproduction of carotenoids is associated with cell growth in the exponential phase, and the average specific growth (μ) in bioreactor is 0.046 h⁻¹ with a maximum yield of 0.19 g cells/L h. The maximum yield of carotenoids (60.0 μg/Lh) is observed at 50-h bioproduction. The conversion factor for total organic carbon (TOC) in cells (Y_X/_SCOT) was 2.97 g/g (0–50 h) and 0.254 g/g (50–100 h), the conversion factor glucose into cells (Y_X/Sglicose) was 0.168 g/g (0–100 h). The specific production of carotenoids (Y_P/_X) was 390 μg of carotenoids per gram of cells, the conversion factor of carbon in the product (Y_P/SCOT) was 107.8 μg/g (0–50 h) and 34.4 μg/g (50–100 h), whereas the factor Y_P/_Sglicose was 69.59 μg/g. The agitation and aeration provided better homogeneity in the culture medium, and hence greater availability of nutrients and oxygen, leading to higher production of carotenoids.     

Sire variation in fatty acid composition of crossbred Wagyu steers and heifers

Effects of sires on lipid composition of subcutaneous adipose tissue and longissimus dorsi muscle were studied using 113 F(1) heifer and steer calves sired by eight Wagyu bulls out of three different cow herds. Wagyu sires were used and grouped as Old (n=6) and New (n =2) sires, respectively, based on the chronological order in which they were imported into the US. Animals were fed a backgrounding diet for 112 days consisting of an 80:20 ratio of roughage:concentrate, then grazed on orchard grass and bluegrass for 84 days, and finished on a 10:90 ratio of roughage:concentrate diet for 231 days in a feedlot. For longissimus dorsi muscle, progeny from Old sires had higher (P<0.05) monounsaturated to saturated fatty acid ratios (MUFA:SFA) than progeny of New sires. There were also differences (P<0.05) among individual sires for polyunsaturated to saturated fatty acid ratio (PUFA:SFA) (0.05-0.08) and MUFA:SFA (1.03-1.21). Progeny of Angus cows at Washington State University (WSU) had lower (P<0.05) MUFA:SFA and lower SFA than progeny of WSU crossbred and commercial cows. Steers had lower (P<0.05) MUFA:SFA and higher (P<0.05) SFA than heifers. For subcutaneous fat, heifers had higher levels (P<0.05) of linoleic acid (C18:2) and PUFA:SFA than the steers. Means for ether extractable fat in longissimus dorsi muscle differed among sires (P<0.05) and ranged from 7.58 to 13.13%. Progeny from WSU Angus cows had higher (P<0.05) ether extractable fat than WSU crossbred and commercial cows. Cholesterol content of longissimus dorsi muscle was not influenced by sire, cow herd or sex (P>0.05).     

Fatty acid composition and sensory characteristics of lamb carcasses from Britain and Spain

Fatty acid composition of intramuscular fat in M. Longissimus was measured in four groups of lambs representing two Spanish breeds: Rasa Aragonesa (RA) and Merino (ME) and two British lamb types: the Welsh Mountain breed (WM), whose carcasses were purchased in Spain and typical early lambs (EL) purchased in Britain. The lambs grown in Spain were concentrate fed and slaughtered at a lighter weight and lower age than those grown in Britain. The British lamb carcasses purchased in Spain were of a similar weight to the Spanish lambs but were lighter than the lamb carcasses purchased in Britain. The British lambs were grass fed. Relationships between fatty acid (FA) composition and sensory attributes were examined following sensory testing of all lambs by both British and Spanish taste panels. The production system was shown to be more important than breed in determining FA composition. British lambs had higher percentages, within total fatty acids, of 18:0, 18:3 (n-3) and long chain polyunsaturated n-3 FA and lower percentages of 18:2 (n-6) and long chain polyunsaturated n-6 FA than Spanish lambs. These differences were due to the different feeding systems used. The amounts of these FA in muscle (mg/100 g) were also different between the British and Spanish groups, not only because the British lambs had more total fat. For both taste panels, odour and flavour intensity were positively correlated with the amounts and percentages of 18:0 and 18:3 and negatively correlated with those of 18:2. This was explained by the fact that both panels gave higher odour and flavour intensity scores to the grass-fed British lamb with high 18:3 levels and lower scores to the concentrate-fed Spanish lamb with high 18:2 levels. However, 18:0 and 18:3 were positively correlated with flavour quality and overall appraisal for the British panel and negatively for the Spanish panel. Conversely, 18:2 was a positive contributor to flavour and overall preference for the Spanish panel and was negative for the British panel. The results show that the production system affects muscle fatty acid composition and the flavour of lamb. However, people's preference is determined to a large extent by their past experience.     

Relationships between meat quality measurements in rabbits fed with three diets of different fat type and content

Two hundred and eighty-four, genetically similar (a three-way cross), young rabbits were fed ad libitum, from weaning, either a commercial diet (group C, ether extract 2.6%) or a diet containing vegetable fat (group V, ether extract, 9.9%) or animal fat (group A, ether extract 11.7%). A principal component (PC) analysis was performed with the variables: ultimate pH at 24 h post mortem measured in the longissimus dorsi (LD) and in the biceps femoris (BF) muscles, colour measured on the surface of the loin, fatty acid composition of perirenal fat, meat fat content of the hind leg, water holding capacity and cooking losses of the meat, and sensory variables determined by a trained panel test. The four first PC explained 62% of the total variation (27, 13, 11 and 11%, respectively). The first PC grouped the fatty acids, the second PC grouped the sensorial variables, and the third and fourth PCs grouped the pHs and the water holding capacity. The projection of the data in the first two PCs showed three separate groups of points. Animals fed with diet V were on the left side of the graph, where the variable C18:2 lies, whereas animals fed with diets A and C lay on the right side of the graph, where the saturated acids were grouped. These were slightly separated by the higher content of oleic acid in the animals fed with diet A. The second PC, where the sensorial variables were grouped, did not separate the animals fed with diets A, V and C. The diets used in this experiment had only a slight influence on the organoleptic characteristics of rabbit meat.