A novel identification method for CPPU-treated kiwifruits based on images.

BACKGROUND: The application of exogenous plant growth regulator, for example forchlorfenuron (CPPU), on kiwifruits has become an important factor that influences kiwifruit economic efficiency and the health development of the kiwifruit industry. Owing to the slight difference in calyx shape between the kiwifruits treated with CPPU (CPPU-treated kiwifruits) and the kiwifruits without CPPU treatment (CPPU-untreated kiwifruits), this study aims to provide a cheap, quick, convenient, and non-destructive method for identifying CPPU-treated kiwifruits based on the images of kiwifruits captured at visible lights. RESULTS: The identification method includes three steps. Firstly, the kiwifruit was extracted from the background by using Otsu algorithm, hole filling operation and 'bwareaopen' function. Secondly, the calyx was extracted by using corrosion, image enhancement, hole filling and closing operations. Finally, the length/width ratio of the minimum enclosing rectangle of calyx region was calculated. The kiwifruit was regarded as a CPPU-treated kiwifruit if the length/width ratio of the rectangle was higher than 1.6. Otherwise, the kiwifruit was regarded as a CPPU-untreated one. The method had the total identification accuracy rate of 90.0% when the kiwifruit images were captured either by utilizing a smartphone at normal lighting condition or by using an image acquisition system. CONCLUSION: The programs run on computer and smartphone were developed, and they could realize kiwifruit identification in 0.6 s and 2 s, respectively. The study makes identifying CPPU-treated kiwifruits in online processing be realizable, and offers a convenient method for kiwifruit consumers. © 2019 Society of Chemical Industry.

HPLC-DAD analysis of florfenicol and thiamphenicol in medicated feedingstuffs.

A simple and reliable method using liquid chromatography with diode array detector was developed for the simultaneous determination of florfenicol and thiamphenicol in medicated feed. The analytes were extracted from the minced feed with methanol and ethyl acetate (1:1, v/v). Next, the extract was further cleaned up by dispersive solid phase extraction using anhydrous magnesium sulfate, PSA and C18 sorbents. Finally, 1 mL of extract was evaporated, the residue resuspended in Milli-Q water, and filtered. The method was validated in-house at medicated levels, in the concentration range 10-300 µg/mL (50-1500 mg/kg). Values of <6.5% and <6.0% were found, respectively, for repeatability and within-laboratory reproducibility. The LODs for the two fenicols were 2.4-5.3 mg/kg, while the LOQs were 3.8-5.6 mg/kg. The expanded uncertainty was estimated to be in the range of 10.0-14.5%, depending on the analyte. Recoveries varied from 81.7% to 97.5%. The methodology was applied to the analysis of animal feedingstuffs collected from poultry and pig farms.

Determination of Chlortetracycline Residues, Antimicrobial Activity and Presence of Resistance Genes in Droppings of Experimentally Treated Broiler Chickens.

Tetracyclines are important antimicrobial drugs for poultry farming that are actively excreted via feces and urine. Droppings are one of the main components in broiler bedding, which is commonly used as an organic fertilizer. Therefore, bedding becomes an unintended carrier of antimicrobial residues into the environment and may pose a highly significant threat to public health. For this depletion study, 60 broiler chickens were treated with 20% chlortetracycline (CTC) under therapeutic conditions. Concentrations of CTC and 4-epi-CTC were then determined in their droppings. Additionally, this work also aimed to detect the antimicrobial activity of these droppings and the phenotypic susceptibility to tetracycline in E. coli isolates, as well as the presence of tet(A), tet(B), and tet(G) resistance genes. CTC and 4-epi-CTC concentrations that were found ranged from 179.5 to 665.8 µg/kg. Based on these data, the depletion time for chicken droppings was calculated and set at 69 days. All samples presented antimicrobial activity, and a resistance to tetracyclines was found in bacterial strains that were isolated from these samples. Resistance genes tet(A) and tet(B) were also found in these samples.

Impacts of feeding preweaned calves milk containing drug residues on the functional profile of the fecal microbiota.

Feeding drug residue-containing milk to calves is common worldwide and no information is currently available on the impact on the functional profile of the fecal microbiota. Our objective was to characterize the functional profile of the fecal microbiota of preweaned dairy calves fed raw milk with residual concentrations of antimicrobials commonly found in waste milk from birth to weaning. Calves were assigned to a controlled feeding trial being fed milk with no drug residues or milk with antibiotic residues. Fecal samples collected from each calf once a week starting at birth, prior to the first feeding in the trial, until 6 weeks of age. Antibiotic residues resulted in a significant difference in relative abundance of microbial cell functions, especially with genes linked with stress response, regulation and cell signaling, and nitrogen metabolism. These changes could directly impacts selection and dissemination of virulence and antimicrobial. Our data also identified a strong association between age in weeks and abundance of Resistance to Antibiotics and Toxic Compounds. Findings from this study support the hypothesis that drug residues, even at very low concentrations, impact the gut microbiota of calves and result in changes in the functional profile of microbial populations.

High occurrence rates of enrofloxacin and ciprofloxacin residues in retail poultry meat revealed by an ultra-sensitive mass-spectrometric method, and antimicrobial resistance to fluoroquinolones in Campylobacter spp.

An ultra-sensitive mass spectrometric confirmation and quantification method for the determination of selected fluoroquinolones-enrofloxacin and its main metabolite ciprofloxacin-was developed and validated in poultry meat samples. The achieved limits of quantification were 1 ng kg-1 for enrofloxacin and 10 ng kg-1 for ciprofloxacin. The analysis of 40 retail poultry samples originating from Estonia, Latvia, Lithuania, Poland and France revealed that 93% of samples contained residues of enrofloxacin in the range from 3.3 to 1126 ng kg-1. Previous studies have shown high levels of antimicrobial resistance to fluoroquinolones, particularly in Campylobacter spp. and various faecal indicators isolated from broiler meat. Consequently, the revealed widespread usage of fluoroquinolones in the poultry industry may result in the further emergence of antimicrobial resistance of Campylobacter in the food chain.

Psychotropic in the environment: risperidone residues affect the behavior of fish larvae.

The ability to avoid and escape from predators are clearly relevant behaviors from the ecological perspective and directly interfere with the survival of organisms. Detected in the aquatic environment, risperidone can alter the behavior of exposed species. Considering the risk of exposure in the early stages of life, we exposed zebrafish embryos to risperidone during the first 5 days of life. Risperidone caused hyperactivity in exposed larvae, which in an environmental context, the animals may be more vulnerable to predation due to greater visibility or less perception of risk areas.

Effects of residual levels of tetracycline on the barrier functions of human intestinal epithelial cells.

Tetracyclines are frequently used in food-producing animals to treat, control, and prevent microbial diseases. Concerns are raised regarding the effects of residual levels of tetracycline, which may be present in the food supply, for emergence of drug-resistance and transfer of antibiotic-resistance gene in intestinal microbiota. In contrast, no information is available regarding the possible effects of residual tetracycline on the gastrointestinal epithelial layer barrier-disruption. This study investigates the outcome of tetracycline treatment on intestinal epithelial cells integrity. Intestinal epithelial cells (T84) were treated at concentrations of 0.015, 0.15, 1.5, 15 and 150 µg/ml for 48 h in an in vitro cell culture model. The permeability study revealed that 15 and 150 µg/ml of tetracycline causes barrier disruption. Whereas the altered mRNA expression of notch-3, notch-4, claudin-2, claudin-8, claudin-10, claudin-15, gap junction alpha 8 and delta 2 and integrin, alpha 3 and alpha L, which are cell-integrity-related genes starts at 1.5 µg/ml tetracycline after 48 h treatment. Translocation of GFP-labeled bacteria from apical to basal comportment provides proof of concept to intestinal barrier disruption. This study is the first to evaluate whether residual concentrations of tetracycline impact epithelial cell integrity.

An update discussion on the current assessment of the safety of veterinary antimicrobial drug residues in food with regard to their impact on the human intestinal microbiome.

The human gastrointestinal tract ecosystem consists of complex and diverse microbial communities that have now been collectively termed the intestinal microbiome. Recent scientific breakthroughs and research endeavours have increased our understanding of the important role the intestinal microbiome plays in human health and disease. The use of antimicrobial new animal drugs in food-producing animals may result in the presence of low levels of drug residues in edible foodstuffs. There is concern that antimicrobial new animal drugs in or on animal-derived food products at residue-level concentrations could disrupt the colonization barrier and/or modify the antimicrobial resistance profile of human intestinal bacteria. Therapeutic doses of antimicrobial drugs have been shown to promote shifts in the intestinal microbiome, and these disruptions promote the emergence of antimicrobial-resistant bacteria. To assess the effects of antimicrobial new animal drug residues in food on human intestinal bacteria, many national regulatory agencies and international committees follow a harmonized process, VICH GL36(R), which was issued by a trilateral organization of the European Union, the USA, and Japan called the International Cooperation on Harmonization of Technical Requirements for Veterinary Medicinal Products (VICH). The guidance describes a general approach currently used by national regulatory agencies and international committees to assess the effects of antimicrobial new animal drug residues in animal-derived food on human intestinal bacteria. The purpose of this review is to provide an overview of this current approach as part of the antimicrobial new animal drug approval process in participating countries, give insights on the microbiological endpoints used in this safety evaluation, and discuss the availability of new information. Copyright © 2016 John Wiley & Sons, Ltd.

Novel bioassay using Bacillus megaterium to detect tetracycline in milk.

Tetracyclines are used for the prevention and control of dairy cattle diseases. Residues of these drugs can be excreted into milk. Thus, the aim of this study was to develop a microbiological method using Bacillus megaterium to detect tetracyclines (chlortetracycline, oxytetracycline and tetracycline) in milk. In order to approximate the limits of detection of the bioassay to the Maximum Residue Limit (100µg/l) for milk tetracycline, different concentrations of chloramphenicol (0, 1000, 1500 and 2000µg/l) were tested. The detection limits calculated were similar to the Maximum Residue Limits when a bioassay using B. megaterium ATCC 9885 spores (2.8×10(8)spores/ml) and chloramphenicol (2000µg/l) was utilized. This bioassay detects 105µg/l of chlortetracycline, 100µg/l of oxytetracycline and 134µg/l of tetracycline in 5h. Therefore, this method is suitable to be incorporated into a microbiological multi-residue system for the identification of tetracyclines in milk.

Modeling the Impact of Ingoing Sodium Nitrite, Sodium Ascorbate, and Residual Nitrite Concentrations on Growth Parameters of Listeria monocytogenes in Cooked, Cured Pork Sausage.

Sodium nitrite has been identified as a key antimicrobial ingredient to control pathogens in ready-to-eat (RTE) meat and poultry products, including Listeria monocytogenes. This study was designed to more clearly elucidate the relationship between chemical factors (ingoing nitrite, ascorbate, and residual nitrite) and L. monocytogenes growth in RTE meats. Treatments of cooked, cured pork sausage (65% moisture, 1.8% salt, pH 6.6, and water activity 0.98) were based on response surface methodology with ingoing nitrite and ascorbate concentrations as the two main factors. Concentrations of nitrite and ascorbate, including star points, ranged from 0 to 352 and 0 to 643 ppm, respectively. At one of two time points after manufacturing (days 0 and 28), half of each treatment was surface inoculated to target 3 log CFU/g of a five-strain L. monocytogenes cocktail, vacuum packaged, and stored at 7°C for up to 4 weeks. Growth of L. monocytogenes was measured twice per week, and enumerations were used to estimate lag time and growth rates for each treatment. Residual nitrite concentrations were measured on days 0, 4, 7, 14, 21, and 28, and nitrite depletion rate was estimated by using first-order kinetics. The response surface methodology was used to model L. monocytogenes lag time and growth rate based on ingoing nitrite, ascorbate, and the residual nitrite remaining at the point of inoculation. Modeling results showed that lag time was impacted by residual nitrite concentration remaining at inoculation, as well as the squared term of ingoing nitrite, whereas growth rate was affected by ingoing nitrite concentration but not by the remaining residual nitrite at the point of inoculation. Residual nitrite depletion rate was dependent upon ingoing nitrite concentration and was only slightly affected by ascorbate concentration. This study confirmed that ingoing nitrite concentration influences L. monocytogenes growth in RTE products, yet residual nitrite concentration contributes to the antimicrobial impact of nitrite as well.

[Development and validation of an analytical method to quantify residues of cleaning products able to inactivate prion]. / Développement et validation d'une méthode de dosage des traces de détergents inactivants totaux du prion.

OBJECTIVES: In this study, a novel analytical method to quantify prion inactivating detergent in rinsing waters coming from the washer-disinfector of a hospital sterilization unit has been developed. The final aim was to obtain an easy and functional method in a routine hospital process which does not need the cleaning product manufacturer services. METHODS: An ICP-MS method based on the potassium dosage of the washer-disinfector's rinsing waters was developed. Potassium hydroxide is present on the composition of the three prion inactivating detergent currently on the French market. The detergent used in this study was the Actanios LDI(®) (Anios laboratories). A Passing and Bablok regression compares concentrations measured with this developed method and with the HPLC-UV manufacturer method. RESULTS: According to results obtained, the developed method is easy to use in a routine hospital process. The Passing and Bablok regression showed that there is no statistical difference between the two analytical methods during the second rinsing step. Besides, both methods were linear on the third rinsing step, with a 1.5ppm difference between the concentrations measured for each method. CONCLUSIONS: This study shows that the ICP-MS method developed is nonspecific for the detergent, but specific for the potassium element which is present in all prion inactivating detergent currently on the French market. This method should be functional for all the prion inactivating detergent containing potassium, if the sensibility of the method is sufficient when the potassium concentration is very low in the prion inactivating detergent formulation.

Dissipation of Fungicide Residues during Winemaking and Their Effects on Fermentation and the Volatile Composition of Wines.

The effects of four fungicides commonly used for the control of fungal diseases in vines and grapes in the course of winemaking were tested. The concentration of fungicide residues was monitored throughout the process to establish their kinetics of dissipation. In all cases the percentages of dissipation were >68%, which shows the detoxificant effect of the winemaking process. On the other hand, the effect of the fungicide residues on the aroma composition of Tempranillo red wines was tested. To evaluate possible modifications on the aroma profile of wines, seven odorant series (ripe fruits, fresh fruits, lactic, floral, spicy, vinous, and herbaceous) were built from the odor activity values (OAVs) obtained for each volatile compound. Ripe fruits and fresh fruits were the major aromatic attributes in all Tempranillo red wines. These two odorant series registered the highest variations in their total OAVs with respect to the control wine, especially with the application of boscalid + kresoxim-methyl into vines, leading to a decrease in the ripe fruit and fresh fruit nuances of the resulting wines. Moreover, when the effect of these fungicides on the aroma of Tempranillo red wines was compared throughout two years (2012 and 2013), wines elaborated from grapes treated in the field with boscalid + kresoxim-methyl in 2013 displayed the highest variation in aroma profile with respect to control wine.

Soil-borne reservoirs of antibiotic-resistant bacteria are established following therapeutic treatment of dairy calves.

We determined if antibiotics residues that are excreted from treated animals can contribute to persistence of resistant bacteria in agricultural environments. Administration of ceftiofur, a third-generation cephalosporin, resulted in a ∼ 3 log increase in ceftiofur-resistant Escherichia coli found in the faeces and pen soils by day 10 (P = 0.005). This resistant population quickly subsided in faeces, but was sustained in the pen soil (∼ 4.5 log bacteria g(-1)) throughout the trial (1 month). Florfenicol treatment resulted in a similar pattern although the loss of florfenicol-resistant E. coli was slower for faeces and remained stable at ∼ 6 log bacteria g(-1) in the soil. Calves were treated in pens where eGFP-labelled E. coli were present in the bedding (∼ 2 log g(-1)) resulting in amplification of the eGFP E. coli population ∼ 2.1 log more than eGFP E. coli populations in pens with untreated calves (day 4; P < 0.005). Excreted residues accounted for > 10-fold greater contribution to the bedding reservoir compared with shedding of resistant bacteria in faeces. Treatment with therapeutic doses of ceftiofur or florfenicol resulted in 2-3 log g(-1) more bacteria than the estimated ID50 (2.83 CFU g(-1)), consistent with a soil-borne reservoir emerging after antibiotic treatment that can contribute to the long-term persistence of antibiotic resistance in animal agriculture.

Adaption of the microbial community to continuous exposures of multiple residual antibiotics in sediments from a salt-water aquacultural farm.

Residual antibiotics from aquacultural farming may alter microbial community structure in aquatic environments in ways that may adversely or positively impact microbially-mediated ecological functions. This study investigated 26 ponds (26 composited samples) used to produce fish, razor clam and shrimp (farming and drying) and 2 channels (10 samples) in a saltwater aquacultural farm in southern China to characterize microbial community structure (represented by phospholipid fatty acids) in surface sediments (0-10 cm) with long-term exposure to residual antibiotics. 11 out of 14 widely-used antibiotics were quantifiable at µg kg(-1) levels in sediments but their concentrations did not statistically differ among ponds and channels, except norfloxacin in drying shrimp ponds and thiamphenicol in razor clam ponds. Concentrations of protozoan PLFAs were significantly increased in sediments from razor clam ponds while other microbial groups were similar among ponds and channels. Both canonical-correlation and stepwise-multiple-regression analyses on microbial community and residual antibiotics suggested that roxithromycin residuals were significantly related to shifts in microbial community structure in sediments. This study provided field evidence that multiple residual antibiotics at low environmental levels from aquacultural farming do not produce fundamental shifts in microbial community structure.

In vivo selection of resistant E. coli after ingestion of milk with added drug residues.

Antimicrobial resistance represents a major global threat to modern medicine. In vitro studies have shown that very low concentrations of drugs, as frequently identified in the environment, and in foods and water for human and animal consumption, can select for resistant bacteria. However, limited information is currently available on the in vivo impact of ingested drug residues. The objective of our study was to evaluate the effect of feeding preweaned calves milk containing antimicrobial drug residues (below the minimum inhibitory concentration), similar to concentrations detected in milk commonly fed to dairy calves, on selection of resistant fecal E. coli in calves from birth to weaning. At birth, thirty calves were randomly assigned to a controlled feeding trial where: 15 calves were fed raw milk with no drug residues (NR), and 15 calves were fed raw milk with drug residues (DR) by adding ceftiofur, penicillin, ampicillin, and oxytetracycline at final concentrations in the milk of 0.1, 0.005, 0.01, and 0.3 µg/ml, respectively. Fecal samples were rectally collected from each calf once a week starting at birth prior to the first feeding in the trial (pre-treatment) until 6 weeks of age. A significantly greater proportion of E. coli resistant to ampicillin, cefoxitin, ceftiofur, streptomycin and tetracycline was observed in DR calves when compared to NR calves. Additionally, isolates from DR calves had a significant decrease in susceptibility to ceftriaxone and ceftiofur when compared to isolates from NR calves. A greater proportion of E. coli isolates from calves in the DR group were resistant to 3 or more antimicrobial drugs when compared to calves in the ND group. These findings highlight the role that low concentrations of antimicrobial drugs have on the evolution and selection of resistance to multiple antimicrobial drugs in vivo.

Ciprofloxacin residues in municipal biosolid compost do not selectively enrich populations of resistant bacteria.

Biosolids and livestock manure are valuable high-carbon soil amendments, but they commonly contain antibiotic residues that might persist after land application. While composting reduces the concentration of extractable antibiotics in these materials, if the starting concentration is sufficiently high then remaining residues could impact microbial communities in the compost and soil to which these materials are applied. To examine this issue, ciprofloxacin was added to biosolid compost feedstock to achieve a total concentration of 19 ppm, approximately 5-fold higher than that normally detected by liquid chromatography-tandem mass spectrometry (LC-MS/MS) (1 to 3.5 ppm). This feedstock was placed into mesh bags that were buried in aerated compost bays. Once a week, a set of bags was removed and analyzed (treated and untreated, three replicates of each; 4 weeks). Addition of ciprofloxacin had no effect on the recovery of resistant bacteria at any time point (P = 0.86), and a separate bioassay showed that aqueous extractions from materials with an estimated 59 ppm ciprofloxacin had no effect on the growth of a susceptible strain of Escherichia coli (P = 0.28). Regression analysis showed that growth of the susceptible strain of E. coli can be reduced given a sufficiently high concentration of ciprofloxacin (P < 0.007), a result that is consistent with adsorption being the primary mechanism of sequestration. While analytical methods detected biologically significant concentrations of ciprofloxacin in the materials tested here, the culture-based methods were consistent with the materials having sufficient adsorptive capacity to prevent typical concentrations of ciprofloxacin residues from selectively enriching populations of resistant bacteria.

High-performance thin-layer chromatography screening of multi class antibiotics in animal food by bioluminescent bioautography and electrospray ionization mass spectrometry.

The world-wide usage and partly abuse of veterinary antibiotics resulted in a pressing need to control residues in animal-derived foods. Large-scale screening for residues of antibiotics is typically performed by microbial agar diffusion tests. This work employing high-performance thin-layer chromatography (HPTLC) combined with bioautography and electrospray ionization mass spectrometry introduces a rapid and efficient method for a multi-class screening of antibiotic residues. The viability of the bioluminescent bacterium Aliivibrio fischeri to the studied antibiotics (16 species of 5 groups) was optimized on amino plates, enabling detection sensitivity down to the strictest maximum residue limits. The HPTLC method was developed not to separate the individual antibiotics, but for cleanup of sample extracts. The studied antibiotics either remained at the start zones (tetracyclines, aminoglycosides, fluoroquinolones, and macrolides) or migrated into the front (amphenicols), while interfering co-extracted matrix compounds were dispersed at hRf 20-80. Only after a few hours, the multi-sample plate image clearly revealed the presence or absence of antibiotic residues. Moreover, molecular information as to the suspected findings was rapidly achieved by HPTLC-mass spectrometry. Showing remarkable sensitivity and matrix-tolerance, the established method was successfully applied to milk and kidney samples.

Insights into the relationship between antimicrobial residues and bacterial populations in a hospital-urban wastewater treatment plant system.

The relationship between antimicrobial residues, antibiotic resistance prevalence and bacterial community composition in hospital effluent and in the receiving wastewater treatment plant was studied. Samples from hospital effluent, raw inflow and final effluent of the receiving wastewater treatment plant were characterized for amoxicillin and ciprofloxacin resistance prevalence, content of heavy metals and antimicrobial residues and bacterial community structure, based on 16S rRNA gene PCR-DGGE analysis. The concentration of fluoroquinolones, arsenic and mercury was in general higher in hospital effluent than in raw inflow, while the opposite was observed for tetracyclines, sulfonamides and penicillin G. The prevalence of ciprofloxacin resistance was significantly higher in hospital effluent than in raw inflow. The concentration of antimicrobial residues was observed to be significantly correlated with the prevalence of antibiotic resistant bacteria and with variations in the bacterial community. Hospital effluent was confirmed as a relevant, although not unique, source of antimicrobial residues and antibiotic resistant bacteria to the wastewater treatment plant. Moreover, given the high loads of antibiotic residues and antibiotic resistant bacteria that may occur in hospital effluents, these wastewater habitats may represent useful models to study and predict the impact of antibiotic residues on bacterial communities.

Detection of maduramycin residues in the tissues of chickens and pheasants by the screening test for antibiotic residues (STAR).

Maduramycin is a coccidiostat authorized as a feed additive in poultry. Council Directive 96/23/EC stipulates that monitoring of foods of animal origin for residues of coccidiostats is mandatory. The aim of the present study was to evaluate the STAR for the screening of maduramycin residues in the tissues of broiler chickens and pheasants. Both animal species were supplied feed medicated with Cygro l% premix according to recommendations for use (5 mg kg(-1) of complete feed). The residues were investigated for a period of 7 days: day 0 (the last day of the administration of maduramycin), days 1-5 (the days of the withdrawal period) and day 6 (the first day after elapse of the withdrawal period). According to STAR the positivity of the sample (the presence of residues of antibacterial substances) is indicated by a zone of inhibition exceeding 2 or 4 mm in width, depending on the test organism. Maduramycin residues were detected only on the plates seeded with the test organism Bacillus stearothermophilus var. calidolactis ATCC 10149. The results showed that there was higher potential for the presence of maduramycin residues in broiler chickens than in pheasants. All chicken tissues (muscle/thigh and breast/gizzard, liver, heart, kidneys, spleen, lungs) were positive for maduramycin (inhibition zones ≥4 mm) not only throughout the withdrawal period, but also even 5 days after elapse of the withdrawal period. In the case of pheasants the positive results were detected in the gizzard, liver, heart, kidneys, spleen and lungs. On day 5 of the withdrawal period no positive results were detected; however, on day 6 the heart and spleen were positive again.

Heat treatment effects on the antimicrobial activity of macrolide and lincosamide antibiotics in milk.

Antibiotic residues in milk can cause serious problems for consumers and the dairy industry. Heat treatment of milk may diminish the antimicrobial activity of these antibiotic residues. This study analyzed the effect of milk processing (60 °C for 30 min, 120 °C for 20 min, and 140 °C for 10 s) on the antimicrobial activity of milk samples fortified with three concentrations of three macrolides (erythromycin: 20, 40 and 80 µg/liter; spiramycin: 100, 200, and 400 µg/liter; and tylosin: 500, 1,000, and 2,000 µg/liter) and one lincosamide (lincomycin: 1,000, 2,000, and 4,000 µg/liter). To measure the loss of antimicrobial activity, a bioassay based on the growth inhibition of Micrococcus luteus was done. The data were analyzed using a multiple linear regression model. The results indicate that treatment at 120 °C for 20 min produces inactivation percentages of 93% (erythromycin), 64% (spiramycin), 51% (tylosin), and 5% (lincomycin), while treatment at 140 °C for 10 s results in generally lower percentages (30% erythromycin, 35% spiramycin, 12% tylosin, and 5% lincomycin). The lowest loss or lowest reduction of antimicrobial activity (21% erythromycin and 13% spiramycin) was obtained by treatment at 60 °C for 30 min.