Fortified Milk Samples Research Articles

In vitro evaluation of milk incorporated with Eryngium billardieri Delar alcoholic extract for antioxidant and antidiabetic potentials.

The present study evaluates the effect of Eryngium billardieri alcoholic extract (EBAE) at weight/volume levels of 0.5% and 1% on some physicochemical, sensory, and microbial properties of cow milk. Results showed that adding EBAE reduced the samples' pH, lightness index (L*), and whitening index (WI). Besides, it increased the levels of acidity, dry matter, viscosity, color indices a* and b*, total phenol content (TPC), total flavonoid content (TFC), antioxidant activity (AA), and enzyme inhibitory activity (EIA) of the fortified milk samples than the control sample (p < .05). During the storage period, a decrease in TPC, TFC, AA, and EIA was observed in all samples (p < .05). There was a positive direct relationship between the health activity of EBAE and its concentration. Moreover, EBAE was able to reduce the total microbial count (TMC) in the milk samples (p < .05). The sample containing 1% w/v of EBAE had the highest TPC, TFC, AA, and EIA but the lowest TMC, and it was highly acceptable in terms of sensory evaluation. Therefore, this sample was selected as the best treatment in this study. The results of this study confirm that EBAE may be used to improve antioxidant activity and beneficial effects in milk.

Milk fortified with chitooligosaccharide–gallic acid conjugate and its liposome: characterisation, storage stability, and in vitro digestibility

SummaryThe effects of chitooligosaccharide–gallic acid conjugate (CGC) and its liposome (CGLP) at two levels (1.25 and 2.50%, w/v) on quality and shelf‐life of milk kept at 4 °C were studied. Microbial counts including total viable, pathogenic and spoilage bacteria were the lowest for CGC‐fortified milk, especially when 2.50% (w/v) was used (CGC‐2.50) than remaining samples (P &lt; 0.05). Lower thiobarbituric acid reactive substances and peroxide values were obtained for CGC‐2.50 than other samples (P &lt; 0.05). However, higher acidity values and decreased pH and lightness were noticed in the aforementioned sample (P &lt; 0.05). Moreover, all fortified milk samples had higher bioactivities during in vitro digestion, in which CGC‐2.50 had the highest bioactivities (P &lt; 0.05). Higher retention of unsaturated fatty acids content in CGC‐2.50 was obtained than other counterparts (P &lt; 0.05). Therefore, CGC efficiently extended milk shelf‐life with high acceptability at 4 °C for a minimum of 6 days, whereas control had a shelf‐life of less than 4 days.

Effect of Cephalosporin Antibiotics on the Activity of Yoghurt Cultures.

The presence of antibiotics in milk is a significant problem affecting the technological safety of dairy products. The aim of the study was to determine the sensitivity of yoghurt cultures to residual levels of selected cephalosporin antibiotics (cephalexin, cefoperazone, cefquinome, cefazolin, and ceftiofur). Five yoghurt cultures were selected containing strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Artificially fortified milk samples (whole pasteurized milk; 85 °C; 3–5 s) with cephalosporins at a concentration of the maximum residue limit were used to evaluate the sensitivity of the yoghurt cultures by monitoring the pH, titratable acidity, and the concentration of selected organic acids (lactic, pyruvic, citric, acetic, orotic, oxalic, formic, uric, and succinic acids) at the end of fermentation (43 °C; 4–5.5 h; pH ≤ 4.6). The titratable acidity was determined by the Soxhlet–Henkel method and the organic acid concentration was monitored by reversed-phase HPLC. Ceftiofur had the greatest effect on the yoghurt culture activity, with a statistically highly significant effect (p < 0.05) on the pH, titratable acidity, and the content of lactic, pyruvic, and acetic acids in all cultures. Other cephalosporins also showed an inhibitory effect on yoghurt metabolism as seen by the evaluation of the lactic and pyruvic acid concentrations.

Monitoring of Aflatoxin M1 in Various Origins Greek Milk Samples Using Liquid Chromatography Tandem Mass Spectrometry

Aflatoxin M1(AFM1), a major metabolite of Aflatoxin B1(AFB1), has been identified as a potential contaminant in dairy products. Because of its possible carcinogenicity, the legislation limits as set by Commission Regulation (EC) No. 1881/2006 are very strict, namely 0.050 μg kg−1 in milk and 0.025 μg kg−1 in infant formulas. To meet these requirements, a sensitive and accurate method was developed, employing liquid chromatography tandem mass spectrometry (UPLC-MS/MS). Ιmmunoaffinity columns (R-Biopharm) were used for sample purification and preconcentration of the analyte of interest. The quantification of AFM1 was conducted using fortified milk samples, while Aflatoxin B2 (AFB2) was used as an internal standard (IS). The method was validated in terms of linearity, precision, trueness, limits of detection and quantification and uncertainty. The performance criteria for the method were evaluated based on European Commission Regulation (EC) No. 401/2006 and its most recent amendment, as well as the suggested criteria for revision by the EU Reference Laboratory for Mycotoxins and Plant Toxins. The recovery was in the range of 77.9–81.0% for all fortification levels (0.025–0.050–0.075 μg kg−1), with RSDR values (Relative Standard Deviation of intermediate precision) ranging from 6.1% to 12%. The method’s detection and quantification limits were 0.0027 μg kg−1 and 0.0089 μg kg−1, respectively. The occurrence of AFM1 was investigated in 40 samples of different animal origin (cow, goat and sheep milk) provided by Greek producers.

A highly sensitive and dual-readout immunoassay for norfloxacin in milk based on QDs-FM@ALP-SA and click chemistry

A dual-readout immunoassay based on QDs-FM@ALP-SA and click chemistry was developed for quick and sensitive detection of norfloxacin (NOR), which is an important fluoroquinolone antibiotic. In the system, the NOR-biotin conjugate (NOR-Biotin) was synthesized by click chemistry for signal transformation, and alkaline phosphatase–labeled streptavidin (ALP-SA) was attached to quantum dot fluorescence microspheres (QDs-FM) by an activated ester method to form QDs-FM@ALP-SA for signal amplification. Here, QDs-FM was a dual-functional carrier: it was used not only as a chemiluminescence signal amplification carrier but also as a fluorescent signal due to its fluorescence character. The NOR antibody was coated on a 96-well chemiluminescence microtiter plate, and NOR-Biotin was bound to the antibody specifically. Then, QDs-FM@ALP-SA was combined with NOR-Biotin to develop a direct competition chemiluminescence/fluorescence immunoassay (dc-CLIA/FIA). The IC50 values were 0.345 and 1.206 ng/mL for dc-CLIA/FIA, respectively. The linear range was 0.013–12.48 ng/mL and 0.042–39.86 ng/mL, respectively. The recovery from the standard fortified blank milk samples was in the range of 86.44%–101.3%. Therefore, this method could be a useful tool for routine screening of NOR residues in milk.

Development of One-Step Non-Solvent Extraction and Sensitive UHPLC-MS/MS Method for Assessment of N-(n-Butyl) Thiophosphoric Triamide (NBPT) and N-(n-Butyl) Phosphoric Triamide (NBPTo) in Milk.

N-(n-butyl) thiophosphoric triamide (NBPT) is a urease inhibitor utilised in urea-based fertilizers. In Ireland, fertilizer treated with NBPT is applied to pasture to mitigate both ammonia and nitrous oxide emissions, but concerns arise as to the potential for residues in milk products. A quick ultrafiltration extraction and ultra-high performance liquid chromatography coupled with mass spectrometry triple quadrupole (UHPLC-MS/MS) quantitation method was developed and validated in this study. The method was applied in the analysis of samples collected from a field study investigating potential transfer of NBPT residues into milk. NBPT and NBPTo residues, were extracted from fortified milk samples and analysed on a UHPLC-MS/MS with recoveries ranging from 74 to 114%. Validation of the UHPLC-MS/MS method at low (0.0020 mg kg−1) and high (0.0250 mg kg−1) concentration levels in line with SANTE/12682/2019 showed overall trueness in the range of 99 to 104% and precision between 1 and 10%, RSD for both compounds. The limit of quantitation (LOQ) was 0.0020 mg kg−1 and other tested parameters (linearity, sensitivity, specificity, matrix effect, robustness, etc.) satisfied acceptance criteria. Stability assessment using spiked samples revealed the compounds were stable in raw and pasteurised milk for 4 weeks at –80 °C storage temperature. Maintaining samples at pH 8.5–9.0 further improved stability. Analysis of 516 milk samples from the field study found that NBPT and NBPTo concentrations were below the LOQ of 0.0020 mg kg−1, thus suggesting very low risk of residues occurring in the milk. The method developed is quick, robust, and sensitive. The method is deemed fit-for-purpose for the simultaneous determination of NBPT and NBPTo in milk.

Rapid and selective extraction of norfloxacin from milk using magnetic molecular imprinting polymers nanoparticles

The magnetic molecular imprinting polymers nanoparticles (MMIPs NPs) for the extraction of norfloxacin (NOR) were prepared by self-polymerization of dopamine onto the surface of (3-Aminopropyl)triethoxysilane (APTES)-modified Fe3O4 NPs. The MMIPs NPs were characterized by FT-IR spectroscopy, XRD, VSM and TEM. The characterization results show that the MMIPs NPs possess superparamagnetic characteristic with the saturated magnetization value of 38.6 emu/g, and the average particle size of about 14.3 nm. Furthermore, the adsorption performance of the MMIPs NPs was evaluated by the static, dynamic, selective adsorption and reusability experiments. The adsorption capacity of MMINs NPs for NOR is 14.2 mg/g with an imprinted factor of 2.1. About 82.4–102.4% of the added NOR (20, 40, 60 μg/mL) were recovered from fortified milk samples with MMIPs NPs as sorbent. Moreover, the MMIPs NPs can be reused for at least six times.

Detection of antibacterial residues in milk by HPLC-DAD and microbial inhibitor tests

The objective of this study was to assess the detection sensitivity of four commercial microbial inhibition assays (MIAs) in comparison with the results by High Pressure Liquid Chromatography with Diode Array Detection (HPLC-DAD) for residues of three tetracyclines, trimethoprim and sulfadiazine in fortified raw milk samples. MIAs showed fairly good results at detecting sulfadiazine residues, whereas concerning tetracyclines and trimethoprim residues, most of the kits showed insufficient results. HPLC analysis revealed high recoveries of examined compounds with detection limits below their respective maximum residue levels (MRLs). The HPLC method was validated determining linearity, accuracy, precision, detection capability and decision limit. Subsequently, both methods were used to monitor several cow and sheep milk samples for the presence of antibacterial agents. Results obtained showed that MIAs could be successfully used for the analysis of different milk types. However, milk spoilage which was observed in some cases could decrease the sensitivity of the kits, a fact that does not seem to influence the effectiveness of HPLC-DAD method.&lt;br /&gt;&lt;br /&gt;

Determination of cholecalciferol (vitamin D3) in bovine milk by dispersive micro-solid phase extraction based on the magnetic three-dimensional graphene-sporopollenin sorbent

In this research, a new magnetic sorbent material composed of three-dimensional graphene aerogel decorated with Fe3O4 nanoparticles attached to hollow sporopollenin exine capsules was synthesized and applied for extraction of vitamin D3 before HPLC-UV analysis. The adsorbent was characterized by FT-IR, SEM, VSM, and zeta potential techniques. The important parameters of the extraction process, including adsorbent dosage, desorption conditions, adsorption time, pH, and salt concentration, were investigated. Under the optimized condition, the method analytical figures of merit were evaluated as follows: linear dynamic range, 10–500 μg L−1; limit of detection, 3.01 μg L−1; determination coefficient (R2), 0.9960; intra-day RSD, 5.28%; and inter-day RSD, 8.17%. The applicability of the method was assessed for the determination of vitamin D3 in different unfortified and fortified bovine milk samples, and the recoveries in the 71.8–113.3% range with the RSDs within 1.4–7.0% were obtained.

Determination of Tetracyclines in Milk with a Molecularly Imprinted Polymer-Based Microtiter Chemiluminescence Sensor

A molecularly imprinted polymer (MIP) capable of recognizing five tetracyclines using minocycline as the template was synthesized for the first time. The MIP was employed as the recognition reagent to prepare a chemiluminescence sensor on a conventional microtiter plate. The light signal was initiated using the highly efficient bis(2,4,6-trichlorophenyl) oxalate-hydrogen peroxide-imidazole chemiluminescence system. After optimization of several appropriate factors, the sensor was employed to determine five tetracyclines in milk. The developed assay contained only one sample-loading step, so each measurement was completed within 12 min. The limits of detection for these analytes were in the range from 0.5 to 2.0 pg/mL, while the recoveries from the fortified milk samples were between 78.1 and 105%. In addition, the sensor was shown to be reusable for up to four measurements. Hence, this sensor has been demonstrated to be a simple, rapid, sensitive, and durable tool for the determination of tetracyclines in animal-derived food.

Fortification of milk with phytosterol and its effect on sensory and physicochemical properties

AbstractPhytosterols are a group of lipophilic steroid alcohols found in plants, which have been shown to lower cholesterol when supplemented in the diet. A commercial phytosterol preparation was added to milk in the form of an oil-in-water emulsion. For the preparation of an emulsion, diacetyl tartaric acid ester of mono- and diglycerides was used as an emulsifier and butteroil was used as a source of fat. Three emulsion formulations, i.e. A (8% phytosterols), B (10% phytosterols) and C (12% phytosterols), were prepared in which the levels of emulsifier (6.5%) and butteroil (10%) were kept constant, and each emulsion was added to milk at a rate of 5% (w/w). Based on sensory evaluation, B-emulsion formulation was selected for fortification of milk. The phytosterol content of the fortified milk determined by reverse-phase high-performance liquid chromatography was 410.8 mg/100 g. No significant loss in the initial content of phytosterol was observed after 1 week of storage. Sensory and physicochemical analyses indicated that significant differences were not observed between control and fortified milk samples up to 7 days of refrigerated storage. The present study suggests that it is feasible to add phytosterol as a functional ingredient in milk in the form of water-soluble emulsion to enhance health benefits of consumers. Two servings of such fortified milk per day provide almost the entire recommended daily requirement of phytosterol.

Detection of β-Lactams and Chloramphenicol Residues in Raw Milk-Development and Application of an HPLC-DAD Method in Comparison with Microbial Inhibition Assays.

The present study was carried out to assess the detection sensitivity of four microbial inhibition assays (MIAs) in comparison with the results obtained by the High Performance Liquid Chromatography with Diode-Array Detection (HPLC-DAD) method for antibiotics of the β-lactam group and chloramphenicol in fortified raw milk samples. MIAs presented fairly good results when detecting β-lactams, whereas none were able to detect chloramphenicol at or above the permissible limits. HPLC analysis revealed high recoveries of examined compounds, whereas all detection limits observed were lower than their respective maximum residue limits (MRL) values. The extraction and clean-up procedure of antibiotics was performed by a modified matrix solid phase dispersion procedure using a mixture of Plexa by Agilent and QuEChERS as a sorbent. The HPLC method developed was validated, determining the accuracy, precision, linearity, decision limit, and detection capability. Both methods were used to monitor raw milk samples of several cows and sheep, obtained from producers in different regions of Greece, for the presence of examined antibiotic residues. Results obtained showed that MIAs could be used effectively and routinely to detect antibiotic residues in several milk types. However, in some cases, spoilage of milk samples revealed that the kits’ sensitivity could be strongly affected, whereas this fact does not affect the effectiveness of HPLC-DAD analysis.

Oxidative stability of alpha‐linolenic acid (ω‐3) in flaxseed oil microcapsules fortified market milk

The oxidative stability of α‐linolenic acid (ALA) was investigated in market milk fortified with flaxseed oil microcapsules. Milk was fortified at levels of 1, 2 and 3 g/100 mL using flaxseed oil powder, and optimised on the basis of sensory scores. No significant difference was observed between the control, 1 and 2 g/100 mL fortified milk samples. Therefore, 2 g/100 mL fortified milk was further evaluated for oxidative stability, fatty acids profile and sensory acceptability during 5 days of storage. The fortified milk was oxidatively stable and sensorially acceptable, retaining ~10.35% ALA, which indicated that 250 mL of milk could meet ~46% of the RDA for ω‐3 fatty acids.

Impact of fortification with iron salts and vitamin A on the physicochemical properties of laboratory‐pasteurised toned milk and bioaccessibility of the added nutrients

The sensory and physicochemical attributes of pasteurised milk fortified with soluble ferric pyrophosphate (FPP) (25 mg/L iron) and vitamin A (2500 IU) were comparable to unfortified control samples. The heat coagulation time (HCT) of fortified milks was comparable to the control and exhibited a type A curve with HCT–pH maxima at the acidic side of the natural pH. The distribution pattern of iron in different milk fractions was similar in control and fortified milk samples. The bioaccessibility of iron, estimated through in vitro digestibility of milk fortified with vitamin A and iron, was found to be slightly higher than that of milk fortified with iron alone.

Validation of a lateral flow test (MRLAFMQ) for the detection of aflatoxin M1 at 50 ng l−1 in raw commingled milk

Aflatoxin M1 contamination in dairy products is a risk when feedstuff contaminated with aflatoxin B1 produced by moulds is consumed by milk-producing animals. Milk can be screened for aflatoxin M1 at the European Union maximum limit of 50 ng l−1 by a lateral flow test, the MRLAFMQ (Aflatoxin M1) Test. The method takes 15 min with no milk dilution or a sample preparation step. The lateral flow assay was validated at the Technology and Food Science Unit of the Institute for Agricultural and Fisheries Research (ILVO-T&V) according to European Union guidelines using fortified raw milk samples. A detection capability of 50 ng l−1 was demonstrated with a false negative rate lower than 2% at 50 ng l−1 and a false positive rate of less than 0.3%. Quantitative readings had a mean bias of +2 to 6 ng l−1 at 50 ng l−1 with a standard deviation of 5–8 ng l−1. Based on the validation results, the test could be considered appropriate for milk screening prior to milk unload at dairies.

Development of a Broad Specific Monoclonal Antibody for Fluoroquinolone Analysis

Fast and sensitive screening of antibiotics is a great need in food quality assurance. A monoclonal antibody specific to fluoroquinolones (FQ) was obtained using ciprofloxacin (CPF) derivant as hapten. The antibody was characterized with broad recognition to CPF (100 %), enrofloxacin (ENF, 73.89 %), norfloxacin (73.57 %), nadifloxacin (67.28 %), danofloxacin (53.09 %), pefloxacin (50.26 %), lomefloxacin (35.66 %), enoxacin (12.40 %), and sarafloxacin (3.23 %). Then, an enzyme linked immunosorbent assay (ELISA) was established. The optimized concentrations of coating antigen and antibody were 0.1 and 0.5 μg/ml, respectively. Various parameters including pH values, ionic strength and the concentration of antibody and antigen were optimized for this assay. The ELISA was found to have the best sensitivity in assay buffer of pH 6 and sodium chloride content 1.6 % (m/v) using CPF as tested compound. Fortified milk samples with CPF and ENF (50 and 250 μg/l) and fortified chicken samples (10 and 50 μg/kg) were analyzed with the proposed measure. The ELISA was examined with recovery range of 94–104 % for milk detection and 93–108 % for chicken detection. The coefficient variation data for intra-assay and inter-assay ranged from 4.24 % to 12.16 %. All results indicate the potential extensive application prospects of this ELISA in FQ monitoring for food safety.

Development of an analytical protocol for the estimation of vitamin D2 in fortified toned milk

A simple and rapid method has been developed for the isolation and determination of vitamin D2 in fortified milk samples by HPLC. The advantage of the proposed method is that sample preparation time is reduced and both crystalline and encapsulated forms of vitamin D2 can be estimated. The developed protocol involves a few extraction steps and requires less amounts of reagents. The HPLC separation of vitamin D2 was carried out on a reverse phase C18 column with photo diode array detector at 254nm. Some additional steps were required for extraction of microencapsulated vitamin D2 for breaking down coating material in comparison to crystalline vitamin D2. The recovery of vitamin D2 in fortified toned milk by the proposed method ranged from 96.46% to 99.05%. Non significant degradation was observed in vitamin D2 during seven days storage and also under light exposure (1485 lux) for 32h.

Effect of β-lactoglobulin A and B whey protein variants on cheese yield potential of a model milk system

Cheese yield mainly depends on the amount and proportion of milk constituents; however, genetic variants of the proteins present in milk may also have an important effect. The objective of this research was to study the effect of the variants A and B of β-lactoglobulin (LG) on cheese yield using a model system consisting of skim milk powder fortified with different levels of a mixture containing α-lactalbumin and β-LG genetic variants (A, B, or A-B) in a 1:2 ratio. Fortified milk samples were subjected to pasteurization at 65°C for 30min. Miniature cheeses were made by acidifying (pH=5.9) fortified milk and incubating with rennet for 1h at 32°C. The clot formed was cut, centrifuged at 2,600×g for 30min at 20°C and drained for determining cheese yield. Cheese-yielding capacity was expressed as actual yield (grams of cheese curd per 100g of milk) and dry weight yield (grams of dried cheese curd per 100g of milk). Free-zone capillary electrophoresis was used for determining β-LG A or B recovery in the curd during rennet-induced coagulation. The presence of β-LG variant B resulted in a significantly higher actual and dried weight cheese yield than when A or A-B were present at levels ≤0.675% of whey protein (WP) addition. Results of free-zone capillary electrophoresis allowed us to infer that β-LG B associates with the casein micelles during renneting, as shown by an increase in the recovery of this variant in the curd when β-LG B was added up to a maximum at 0.45% (equivalent to 0.675% WP). In general, actual or dried weight cheese yield increased as WP addition was increased from 0.225 to 0.675%. However, when WP addition ranged from 0.675 to 0.90%, a drastic drop in cheese yield was observed. This behavior may be because an increase in the aggregation of casein micelles with a concomitant inclusion of whey protein in the gel occurs at low levels of WP addition, whereas once the association of WP with the casein micelles reach a saturation point at addition levels higher than 0.675%, rearrangements of the gel network result in larger whey expulsion and syneresis. This knowledge is expected to be useful to maximize cheese yield and optimize processing conditions during cheese and cheese analogs manufacturing.

Semiautomated solid-phase extraction manifold with a solvent-level sensor

A semiautomated solid-phase extraction manifold for multiple extractions is presented. The manifold utilizes commercial solid-phase syringe cartridges and automatically introduces and elutes all the solvents during the extraction, reducing the typical workload and stress of the analyst. The manifold consists of a peristaltic pump with solenoid valves in a flow circuit that contains transmissive photomicrosensors. The photomicrosensors were used to control the solvent dispenser and the solvent level inside the cartridge. As solvent-level sensors, the photomicrosensors determined the exact time the solvent reached the top frit to avoid sorbent drying and accurately perform the solvent exchange. The repeatability of the manifold to introduce a particular volume of solvent into the cartridges was measured, and the precisions were between 0.05 and 2.89% (RSD). To evaluate the manifold, the amount of two fluoroquinolones in a fortified blank milk sample was determined. The results of the intra- and inter-day precision of multiple extractions from the fortified milk samples resulted in precisions better than 9.0% (RSD) and confirmed that the arrangement of the semiautomated manifold could adequately be used in solid-phase extraction with commercial cartridges.

Comparison of detection sensitivity of five microbial inhibition tests for the screening of aminoglycoside residues in fortified milk

The assessment of detection sensitivity of five microbial inhibition tests (MITs), STAR (screening test for antibiotic residues) with the test strain Bacillus subtilis BGA, Delvotest&lt;sup&gt;&amp;reg; &lt;/sup&gt;S&amp;nbsp; P-NT, Total Antibiotics, Kalidos TB, and Kalidos MP with the test strain Bacillus stearothermophilus var. calidolactis to five aminoglycosides (AMGs), gentamicin, neomycin, streptomycin, kanamycin, and spectinomycin in fortified milk samples were studied. The sensitivity of MITs to AMGs was evaluated on the basis of experimental determination of detection limits (LODs) of MITs for AMGs. The LODs of these tests were compared with the maximum residue limits (MRLs) established for milk by the Commission Regulation (EU) No. 37/2010. LODs of STAR for AMGs in fortified milk samples were at the levels of MRL for neomycin (1.50 &amp;micro;g/g), gentamicin (0.10&amp;nbsp;&amp;micro;g/g), streptomycin (0.20 &amp;micro;g/g) and kanamycin (0.15 &amp;micro;g/g). Spectinomycin (0.20 &amp;micro;g/g) was not detected at the level of MRL. The LODs determined by Delvotest&lt;sup&gt;&amp;reg; &lt;/sup&gt;SP-NT, Total Antibiotics and Kalidos MP were comparable, but only gentamicin and neomycin were reliably detected at the levels of MRL. Kalidos TB was more sensitive to AMGs than Delvotest&lt;sup&gt;&amp;reg; &lt;/sup&gt;SP-NT, Total Antibiotics and Kalidos MP. Gentamicin, neomycin and streptomycin were detected at the levels of MRL. &amp;nbsp;