Antimicrobials in Eisenia fetida earthworms: A comprehensive study from method development to the assessment of uptake and degradation.

In this work, an accurate analytical method was developed for the simultaneous analysis of twenty-seven antimicrobials (AMs) in earthworms using liquid chromatography coupled to a triple quadrupole mass spectrometry detector (UHPLC-MS/MS). Adequate apparent recoveries (80-120 %) and limits of quantification (LOQ) (1 µg·kg-1 - 10 µg·kg-1) were obtained, with the exception of norfloxacin (34 µg·kg-1). The method was applied to evaluate the accumulation of sulfamethazine (SMZ) and tetracycline (TC) in earthworms after performing OECD-207 toxicity test, in which Eisenia fetida (E. fetida) organisms were exposed to soils spiked with 10 mg·kg-1, 100 mg·kg-1 or 1000 mg·kg-1 of SMZ and TC, individually. The results confirmed the bioaccumulation of both AMs in the organisms, showing a greater tendency to accumulate SMZ since higher bioconcentration factor values were obtained for this compound at the exposure concentrations tested. In addition, the degradation of both AMs in both matrices, soils and earthworms was studied using liquid chromatography coupled to a q-Orbitrap high resolution mass spectrometry detector. Thirteen transformation products (TPs) were successfully identified, eight of them being identified for the first time in soil/earthworm (such as 4-Amino-3-chloro-n-(4,6-dimethylpyrimidin-2-yl)benzenesulfonamide or 4-(dimethylamino)-1,11,12a-trihydroxy-6,6-dimethyl-3,7,10,12-tetraoxo-3,4,4a,5,5a,6,7,10,12,12a-decahydrotetracene-2-carboxamide, among others) and their formation/degradation trend over time was also studied. Regarding the biological effects, only SMZ caused changes in earthworm growth, evidenced by weight loss in earthworms exposed to concentrations of 100 mg·kg-1 and 1000 mg·kg-1. Riboflavin decreased at all concentrations of SMZ, as well as at the highest concentration of TC. This indicates that these antibiotics can potentially alter the immune system of E. fetida. This research represents a significant advance in improving our knowledge about the contamination of soil by AM over time. It investigates the various ways in which earthworms are exposed to AMs, either by skin contact or ingestion. Furthermore, it explores how these substances accumulate in earthworms, the processes by which earthworms break them down or metabolise them, as well as the resulting TPs. Finally, it examines the potential effects of these substances on the environment.

A wearable gloved sensor based on fluorescent Ag nanoparticles and europium complexes for visualized assessment of tetracycline in food samples.

The abuse of tetracycline antibiotics leads to accumulating residues in the human body, seriously affecting human health. Establishing a sensitive, efficient, and reliable method for qualitative and quantitative detection of tetracycline (TC) is necessary. This study integrated silver nanoclusters and europium-based materials into the same nano-detection system to construct a visual and rapid TC sensor with rich fluorescence color changes. The nanosensor has the advantages of a low detection limit (10.5 nM), high detection sensitivity, fast response, and wide linear range (0-30 µM), which can meet the analysis requirements of different types of food samples. In addition, portable devices based on paper and gloves were designed. Through the smartphone's chromaticity acquisition and calculation analysis application (APP), the real-time rapid visual intelligent analysis of TC in the sample can be realized, which guides the intelligent application of multicolor fluorescent nanosensors.

Response of submerged macrophytes and biofilms to coexisting azithromycin and tetracycline: Antibiotic resistance genes removal, toxicity assessment and microbial properties.

Antibiotics, such as azithromycin (AZ), tetracycline (TC), and their related antibiotic resistance genes (ARGs), create serious ecological risks to aquatic organisms. This study examined the response mechanisms of submerged macrophytes and periphytic biofilms to a mixture of AZ and TC pollution and determined the antibiotic removal efficiencies and fate of ARGs. The results showed that the plant-biofilm system had a significant capacity for removing both single and combined antibiotics with removal efficiencies of 93.06% ∼99.80% for AZ and 73.35% ∼97.74% for TC. Higher ARG (tetA, tetC, tetW, ermF, ermX, and ermB) abundances were observed in the biofilm, and subsequent exposure to the antibiotic mixture increased the abundances of these genes. Both single and combined antibiotics triggered antioxidant stress, but antagonistic effects were induced only with mixed AZ and TC exposure. Furthermore, the antibiotics changed the structural characteristics of extracellular polysaccharides and induced alterations in the structure of the biofilm microbial community. Increased N-acylated-l-homoserine lactone confirmed alternations in microbial quorum-sensing. The results extend the understanding of the fate of antibiotics and ARGs when aquatic plants and biofilms are exposed to antibiotic mixtures, as well as the organism's response mechanisms.

Assessment of agricultural waste biochars for remediation of degraded water-soil environment: Dissolved organic carbon release and immobilization of impurities in one- or two-adsorbate systems.

This paper presents a method of agricultural waste management - the production of two biochars (BC) from potato and raspberry stems. It defines the potential of these materials for remediation of degraded water and soil environments. The performed study included analyses of BC physicochemistry, dissolved organic carbon (DOC) release and ability to immobilize copper (Cu), tetracycline (TC) and carboxin (CB) in one- and two-adsorbate systems. The BCs were obtained with pyrolysis at 600 °C for 30 min in a nitrogen atmosphere. Their DOC was predominantly constituted of substances with large molecular weights and high aromaticity, meaning that both BCs can be safely applied as soil additives. Potato-biochar (P-BC) had a more developed surface than raspberry-biochar (R-BC). The specific surface area (SBET) of P-BC was 122 m2/g, whilst of R-BC was 87 m2/g. As a result, the efficiency of impurity adsorption in the one-adsorbate systems was higher for P-BC (61.75% for Cu, 73.84% for TC, and 54.43% for CB). In the two-adsorbate systems, organic impurities improved the immobilization of heavy metal ions on BCs. The efficiency of Cu adsorption on P-BC when TC was present was 88.29%. Desorption of Cu from BC was highest using HCl, whilst that of TC and CB was highest using NaOH. Maximum desorption was observed in a two-adsorbate system with TC + CB (up to 63.6% for TC). These results confirmed that potato and raspberry stems can be used to produce highly effective BCs with large application potential, especially for remediation of degraded soils and polluted waters.

Tetracyclines in pasteurized milk commercialized in São Paulo, Brazil: occurrence and dietary risk assessment based on single and combined exposure with polyether ionophores.

Tetracyclines and polyether ionophores are veterinary drugs frequently used in animals reared for food production. Despite the benefits, residues can be harmful for human health if levels are higher than those considered safe. The aim of this study was to evaluate, by LC-MS/MS, tetracyclines residues in pasteurised milk samples marketed in São Paulo state, Brazil and estimate the exposure of the adult/adolescent/children population. Within the eight antibiotics monitored two were detected, tetracycline and 4-epitetracycline, found in 22.5% and 12.5% of the samples, respectively. Since the same samples had been previously tested for polyether ionophore residues, risk assessment based on dietary exposure to tetracyclines when considered individually and in combination with the ionophores was performed. The daily intakes were below the acceptable daily intake (ADI) for tetracycline. Considering the levels of monensin reported in a previous study, the combined exposure with tetracycline and 4-epi-tetracycline also indicated low potential health concerns.

Health risk assessment of antimicrobial residues in sheep carcasses marketed in Kuwait.

A total of 450 samples comprising 150 each of muscles, livers and kidneys were collected from 150 sheep carcasses in Kuwait and tested by both Rapid Premi®Test kits for rapid detection of antimicrobial-positive samples, and the High-performance liquid chromatography (HPLC) for residual determination of amoxicillin, tetracycline, oxytetracycline, and tylosin. Premi test revealed that 82%, 64% and 100% of the muscle, liver and kidney samples examined, respectively were positive for antimicrobials. HPLC analysis revealed mean concentrations (µg/kg) of 45.26, 148.17, 103.18, and 71.80 for amoxicillin, oxytetracycline, tetracycline, and tylosin respectively in muscles; 64.43, 263.15, 177.04, and 112.94, respectively in livers; and 53.12, 368.21, 196.40, and 138.63, respectively in kidneys. Although many samples exceeded the maximum residue limit (MRL) of Codex Alimentarius, the assessment of dietary exposure to the antimicrobials tested through consumption of sheep meat and organs did not constitute any health hazards in the different age groups of Kuwaiti population.

Sulfonamide and tetracycline in landfill leachates from seven municipal solid waste (MSW) landfills: Seasonal variation and risk assessment.

Antibiotics have received increased attention as emerging contaminants due to their toxicity and potential risk. Landfills serve as one of the important reservoirs of antibiotics. The antibiotics in landfills leaching to nearby environment by leachate may threat ecosystem health. The present study aimed to evaluate the levels of tetracyclines (TCs) and sulfonamides (SAs) in seven Chinese Municipal Solid Waste (MSW) landfill leachates over two years (2017-2018). Seven target antibiotics, TC, oxytetracycline (OTC), doxycycline (DXC), sulfonamide sulfadiazine (SD), sulfamerazine (SM), sulfamethazine (SMX), and sulfamethoxazole (SMT), were detected in 56 landfill leachate samples. Among these, SMT had the highest mean concentration at 654 ng/L (n = 45), followed by OTC (219.58 ng/L, n = 47), and SD (209.98 ng/L, n = 49). The temporal trend showed that antibiotic concentrations were higher in 2017 than in 2018. Furthermore, physicochemical properties were significantly correlated with SAs (p < 0.05), whereas no significant correlation was found for TCs. Seasonal variation analysis revealed that antibiotic levels were higher in spring and winter compared to summer and fall seasons, which might be attributed to the higher waterfall levels in these seasons. Risk assessment revealed that SAs (SM, SMX, SMT) are associated with high risk, and the RQs follow the order of: SMX > SMT > SM. In contrast, TCs had insignificant risk. The findings of this two-year comprehensive monitoring project have produced positive results regarding antibiotic pollution at landfill sites, which can be applied to antibiotics management in landfill and further ensure public health.

Toxicity assessment and microbial response to soil antibiotic exposure: differences between individual and mixed antibiotics.

Increasing amounts of antibiotics are introduced into soils, raising great concerns on their ecotoxicological impacts on the soil environment. This work investigated the individual and joint toxicity of three antibiotics, tetracycline (TC), sulfonamide (SD) and erythromycin (EM) via a whole-cell bioreporter assay. TC, SD and EM in aqueous solution demonstrated cytotoxicity, whilst soil exposure showed genotoxicity, indicating that soil particles possibly affected the bioavailability of antibiotics. Toxicity of soils exposed to TC, SD and EM changed over time, demonstrating cytotoxic effects within 14-d exposure and genotoxic effects after 30 days. Joint toxicity of TC, SD and EM in soils instead showed cytotoxicity, suggesting a synergetic effect. High-throughput sequencing suggested that the soil microbial response to individual antibiotics and their mixtures showed a different pattern. Soil microbial community composition was more sensitive to TC, in which the abundance of Pseudomonas, Pirellula, Subdivision3_genera_incertae_sedis and Gemmata varied significantly. Microbial community functions were significantly shifted by EM amendments, including signal transduction mechanisms, cytoskeleton, cell wall/membrane/envelope biogenesis, transcription, chromatin structure and dynamics, and carbohydrate transport and metabolism. This work contributes to a better understanding of the ecological effects and potential risks of individual and joint antibiotics on the soil environment.

Computational and experimental assessment of pH and specific ions on the solute solvent interactions of clay-biochar composites towards tetracycline adsorption: Implications on wastewater treatment.

Experimental and computational investigations have been conducted in this study to assess the influence of municipal waste pyrolyzed biochar impregnated clay composites on antibiotic removal as a material for wastewater treatment and simultaneous value-addition for waste. The surface potential (zeta potential) of the pristine biochar and composite samples are found to be within the range ~10 to ~ -40 mV in the pH range 2-10. The presence of different inorganic salt solutions influences the electrophoretic mobility of the dispersed phase in a suspension, as well as its zeta potential. In addition of Na+ salt solutions, the Na+ ions undergo electrostatic interaction with the negatively charged biochar samples and form a double layer at the interface of biochar and ionic salt solution. Molecular dynamics simulations have been employed to understand experimental findings, ions adsorption and solute-solvent interactions at the molecular level of two biochar B7 (seven benzene rings, one methoxy, one aldehyde and two hydroxyls groups) and B17 (seventeen benzene rings, one methoxy, two hydroxyls and two carboxylic acid groups) in salts aqueous solutions. The results confirm that hydroxyls and carboxylate groups of biochar are responsible for solute-solvent interactions. Successful removal of tetracycline antibiotics is observed with 26 mg/g maximum adsorption capacity with montmorillonite biochar composite. This study confirms that interactions between amide and hydroxyl groups of tetracycline with hydroxyl and carboxylate groups of biochar play the key role in the adsorption process. The solution pH and presence of different background electrolytes effectively influence the process of solute-solvent interactions as well as adsorption efficacy towards tetracycline adsorption.

The residual tetracycline in pharmaceutical wastewater was effectively removed by using MnO2/graphene nanocomposite.

The object of this study was to remove the tetracycline (TC) residue in pharmaceutical wastewater after flocculation treatment. MnO2/graphene nanocomposite was synthesized by an in situ hydrothermal method and its TC removal rate was up to 99.4%. This nanocomposite had excellent water solubility. More importantly, the introduction of MnO2 nanorods allowed the avoidance of excessive stacking of treated graphene sheets during the adsorption process, which made the TC molecules to have more opportunities to make contact with the adsorbents. In order to eliminate the interference factors, the adsorption isotherm, kinetics, thermodynamics and mechanism were all studied in TC aqueous solution. The influence of solution pH, contact time, MnO2 loading amount, temperature and solution concentration on the adsorption process were also assessed. The main adsorption mechanism contributed to the complexation of Mn(IV) and π-π interactions of the benzene ring structure on treated graphene sheets with TC molecules.

Evaluation of animal feeds for the presence of three important antibiotic classes in Bhutan.

OBJECTIVES: An alarming rate of emergence of antimicrobial resistance (AMR) globally has urged Bhutan to take firm steps towards antibiotic stewardship through rational use in humans, animals and agriculture. This study aimed to investigate the use of three important antibiotic classes in animal feeds in Bhutan. METHODS: Thirteen samples were selected randomly from a pool of feed samples submitted by bidders for Annual Animal Feed Tender (fiscal year 2015-2016) of the Department of Livestock. The 13 samples were tested for nitrofurans and their metabolites, tetracyclines and sulphonamides. The objectives of the study were to evaluate the feed samples for the presence of three antibiotic classes and, if present, to understand the level of presence (mg/kg) in terms of AMR emergence and spread in a Bhutanese context. RESULTS: Two feed samples, belonging to Indian Feed Agents, tested positive for oxytetracycline at levels of 0.072mg/kg and 0.037mg/kg, respectively. Compared with the Feed Additive Compendium recommended dose (10-50g/ton) of oxytetracycline for poultry, the levels found in these two feeds were low. Samples testing positive for antibiotics were rejected in the bidding procedure as being medicated at prophylactic levels and could be the cause of emergence and spread of AMR in the country. CONCLUSION: Although the samples containing antibiotics were rejected by the Department of Livestock for use in government farms, some farmers may still use medicated feeds. Therefore, more screening needs to be conducted to further confirm and to prevent antibiotic abuse in animal feed in Bhutan.

Assessment of veterinary drugs in plants using pharmacokinetic approaches: The absorption, distribution and elimination of tetracycline and sulfamethoxazole in ephemeral vegetables.

The present study was carried out to demonstrate novel use of pharmacokinetic approaches to characterize drug behaviors/movements in the vegetables with implications to food safety. The absorption, distribution, metabolism and most importantly, the elimination of tetracycline (TC) and sulfamethoxazole (SMX) in edible plants Brassica rapa chinensis and Ipomoea aquatica grown hydroponically were demonstrated and studied using non-compartmental pharmacokinetic analysis. The results revealed drug-dependent and vegetable-dependent pharmacokinetic differences and indicated that ephemeral vegetables could have high capacity accumulating antibiotics (up to 160 µg g-1 for TC and 38 µg g-1 for SMX) within hours. TC concentration in the root (Cmax) could reach 11 times higher than that in the cultivation fluid and 3-28 times higher than the petioles/stems. Based on the volume of distribution (Vss), SMX was 3-6 times more extensively distributed than TC. Both antibiotics showed evident, albeit slow elimination phase with elimination half-lives ranging from 22 to 88 hours. For the first time drug elimination through the roots of a plant was demonstrated, and by viewing the root as a central compartment and continuous infusion without a loading dose as drug administration mode, it is possible to pharmacokinetically monitor the movement of antibiotics and their fate in the vegetables with more detailed information not previously available. Phyto-pharmacokinetic could be a new area worth developing new models for the assessment of veterinary drugs in edible plants.

Assessment of oxytetracycline and tetracycline antibiotics in manure samples in different cities of Khuzestan Province, Iran.

Tetracyclines (TCs), a class of antibiotics with a broad spectrum, are the most frequently used antibiotics in animal production. The major concern is that the widespread use of the antibiotics may lead to the emergence of new strains of bacteria that are resistant to these antibiotics. The objective of this study was to determine the residual levels of oxytetracycline and tetracycline in 80 animal manure samples that were collected from the livestock and poultry feedlots in Khuzestan Province. The residual levels of the antibiotics in the samples were extracted by using solid-phase extraction (SPE) method and subsequently were measured by liquid chromatography. Recoveries from the spiked poultry manure samples ranged from 65 to 113% for tetracycline and 86 to 132% for oxytetracycline. Relative standard deviations of the recoveries were less than 5.7% within the same day. Method detection limit (MDL) measured for oxytetracycline and tetracycline in the manure were 0.011 and 0.01 mg/kg, respectively. Analysis of the collected 50 chickens and 30 cow manure samples showed that the highest concentration of tetracycline was related to Behbahan City (5.36 mg/kg) and the lowest concentration was detected for Ramhormoz (0.05 mg/kg). The highest and lowest concentrations of oxytetracycline were respectively observed for Behbahan (13.77 mg/kg) and Ramhormoz (0.047 mg/kg). Based on the results, in chicken manure, there was significant statistical difference between the residual TC concentrations among five cities (p(value) < 0.05). However, no significant relationship was observed between oxytetracyclin (OTC) residual concentrations among five cities (p(value) > 0.05).

In Vivo Transmission of an IncA/C Plasmid in Escherichia coli Depends on Tetracycline Concentration, and Acquisition of the Plasmid Results in a Variable Cost of Fitness.

IncA/C plasmids are broad-host-range plasmids enabling multidrug resistance that have emerged worldwide among bacterial pathogens of humans and animals. Although antibiotic usage is suspected to be a driving force in the emergence of such strains, few studies have examined the impact of different types of antibiotic administration on the selection of plasmid-containing multidrug resistant isolates. In this study, chlortetracycline treatment at different concentrations in pig feed was examined for its impact on selection and dissemination of an IncA/C plasmid introduced orally via a commensal Escherichia coli host. Continuous low-dose administration of chlortetracycline at 50 g per ton had no observable impact on the proportions of IncA/C plasmid-containing E. coli from pig feces over the course of 35 days. In contrast, high-dose administration of chlortetracycline at 350 g per ton significantly increased IncA/C plasmid-containing E. coli in pig feces (P < 0.001) and increased movement of the IncA/C plasmid to other indigenous E. coli hosts. There was no evidence of conjugal transfer of the IncA/C plasmid to bacterial species other than E. coli. In vitro competition assays demonstrated that bacterial host background substantially impacted the cost of IncA/C plasmid carriage in E. coli and Salmonella. In vitro transfer and selection experiments demonstrated that tetracycline at 32 µg/ml was necessary to enhance IncA/C plasmid conjugative transfer, while subinhibitory concentrations of tetracycline in vitro strongly selected for IncA/C plasmid-containing E. coli. Together, these experiments improve our knowledge on the impact of differing concentrations of tetracycline on the selection of IncA/C-type plasmids.

Toxicity of five antibiotics and their mixtures towards photosynthetic aquatic organisms: implications for environmental risk assessment.

The individual and combined toxicities of amoxicillin, erythromycin, levofloxacin, norfloxacin and tetracycline have been examined in two organisms representative of the aquatic environment, the cyanobacterium Anabaena CPB4337 as a target organism and the green alga Pseudokirchneriella subcapitata as a non-target organism. The cyanobacterium was more sensitive than the green alga to the toxic effect of antibiotics. Erythromycin was highly toxic for both organisms; tetracycline was more toxic to the green algae whereas the quinolones levofloxacin and norfloxacin were more toxic to the cyanobacterium than to the green alga. Amoxicillin also displayed toxicity to the cyanobacterium but showed no toxicity to the green alga. The toxicological interactions of antibiotics in the whole range of effect levels either in binary or multicomponent mixtures were analyzed using the Combination Index (CI) method. In most cases, synergism clearly predominated both for the green alga and the cyanobacterium. The CI method was compared with the classical models of additivity Concentration Addition (CA) and Independent Action (IA) finding that CI could accurately predict deviations from additivity. Risk assessment was performed by calculating the ratio between Measured Environmental Concentration (MEC) and the Predicted No Effect Concentration (PNEC). A MEC/PNEC ratio higher than 1 was found for the binary erythromycin and tetracycline mixture in wastewater effluents, a combination which showed a strong synergism at low effect levels in both organisms. From the tested antibiotic mixtures, it can be concluded that certain specific combinations may pose a potential ecological risk for aquatic ecosystems with the present environmentally measured concentrations.

Determination of tetracycline residues in chicken meat by liquid chromatography-tandem mass spectrometry.

Analysis of residual levels of tetracyclines (TCs) in chicken meat was performed using a validated liquid chromatography coupled with a tandem mass spectrometry (LC-MS/MS) technique. Overall, the recoveries for TCs ranged from 56.9% to 101.2%, with standard deviations of 4.5-13.2%. Detection limits ranged from 7.9 to 14.6 µg kg⁻¹. In four of 60 samples, doxycycline (DXC) was determined in a range from 19.9 to 35.6 µg kg⁻¹; and in one sample tetracycline was detected at 17.2 µg kg⁻¹. Chlortetracycline (CTC) and oxytetracycline (OTC) were not detected in any of the tested samples. This study indicates that chicken meat sold in Bursa, Turkey, contained some residues of TCs. Therefore, stricter regulations for the use of antibiotics in the poultry industry and the monitoring of drug residues in chicken meat prior to marketing are needed. Finally, this method has been applied successfully for the confirmation of TCs in chicken meat.

Dietary exposure assessment of streptomycin and tetracycline in food of animal origin on the Croatian market.

Residual antibacterials in food constitute a risk to human health, particularly because they can contribute to the transmission of antibiotic-resistant pathogenic bacteria through the food chain. This paper presents dietary exposure assessment of streptomycin and tetracycline, based on combining food consumption data in Croatia with data on the concentration of veterinary drugs in analysed samples of food of animal origin. According to the median values, the estimated daily intake of streptomycin and tetracycline through food is 11.9 and 0.7 µg/person/day, respectively. The largest contribution to streptomycin intake comes from meat (4.8 µg/person/day, i.e. 41%), but milk is the largest source for tetracycline (0.3 µg/person/day, i.e. 46%). The estimated dietary exposure to these veterinary drugs does not exceed relevant toxicological reference values and the level of exposure is assessed to be acceptable.

Assessment of tetracycline, lead and cadmium residues in frozen chicken vended in Lagos and Ibadan, Nigeria.

This study determined the levels of tetracycline and heavy metals (lead and cadmium) levels in frozen chicken. One hundred frozen chicken muscle samples were sourced from major markets in Lagos and Ibadan (fifty samples each). The samples were analyzed using high power liquid chromatography (HPLC) for tetracycline residue determination while atomic absorption spectroscopy (AAS) was used to determine the levels of lead and cadmium residues in the samples. Mean concentrations of tetracycline residue levels in the frozen chicken sampled ranged from 1.1589-1.0463ppm which is higher than the maximum residue limit set by international food safety agencies. Pb contents were higher in chicken muscles sampled from markets in Ibadan (0.0227 +/- 0.0069 microg dL(-2)) than Lagos (0.0207 +/- 0.0082 microg dL(-1)), while Cd levels were 0.0013 microg dL(-1) higher than in the Lagos samples (0.0065 +/- 0.0026 microg dL(-1)). These values were within maximum residue limits. There were no significant differences (p < 0.05) in levels of tetracycline, lead and cadmium levels from the two market locations (Lagos and Ibadan) and parts (wings and thigh muscles). However, significant differences occurred in tetracycline and Pb levels in frozen chicken sourced from Cotonou. Though not significant, tetracycline contents in the thigh muscles of the frozen chicken samples was higher than that of the wings muscles and this was attributed to the site of administration of antibiotic injection and failure to observe the pre-slaughter withdrawal period by the farmers. This study is of public health importance as the presence of these residues above the maximum residue limit in frozen chicken predisposes consumers to drug resistance, allergic reactions and poisoning as a result of toxicity.

Risk assessment of streptomycin and tetracycline residues in meat and milk on Croatian market.

In addition to biological hazards like bacteria, viruses, parasites, the occurrence of chemical hazards is another characteristic of modern food production. The use of veterinary medicines in intensive production of animals is conditio sine qua non so it is impossible to avoid in full the presence of their residues in food. This paper presents quantitative risk assessment of streptomycin and tetracycline based on acceptable daily intake, daily consumption of milk and meat in Croatia (0.222 and 0.126 kg/person, respectively) and residues of these two veterinary drugs in this type of food. The median value for streptomycin in milk and meat was 11.50 and 38.00 µg/kg, respectively (milk: average: 15.57 µg/kg; range from 0 to 73.82 µg/kg; meat: average 44.14 µg/kg; range from 0 to 278.35 µg/kg). The median value for tetracycline in milk and meat was 1.50 µg/kg (milk: average 1.5 µg/kg; range, from 0 to 4.26 µg/kg; meat: average 1.62 µg/kg; range from 0 to 5.35 µg/kg). Based on the median value it can be concluded that the estimated daily intake of streptomycin and tetracycline through milk and meat in Croatia is low (streptomycin: 7.33 µg/person/day; tetracycline: 0.52 µg/person/day), and the risk is assessed as negligible.

[Spectral control of metal admixtures in tetracycline].

Analysis of circular dichroism spectra made it possible to offer a method for estimation of tetracycline solutions contamination with metal ions. By its sensitivity the method is much superior to the spectrophotometric one used at present for determination of the antibiotic purity. In the latter method formation of complexes with metals is traced by batochromic displacement of the absorption spectra. The new method is rapid, relatively selective and requires comparatively small quantities of the substance for the analysis, which provides its use under both laboratory and manufacture conditions. The method is based on identification of the circular dichroism spectra of tetracycline complexes with metals in the long wavelength region. The presence of the circular dichroism concervative bands with strictly defined extremums in the spectra of tetracycline low acid solutions contaminated by multiply charged metal ions allowed vs. the circular dichroism spectra of pure tetracycline sample to conclude that the solution contained admixtures and to suggest their nature. It was shown that the charge, ion radius and tetracycline:metal relation were the factors defining the mark and location of the dichroism band extremums. At lambda(extr)-410-415 nm the tetracycline complexes with light metal ions such as Mg2+, Al3+ and Ca2+ were detected by the circular dichroism negative band in the spectra, while the complexes with heavy metal ions such as Sc3+, Sr3+, Cu3+, Cd3+, Ba2+, Y3+ and the cerium subgroup lanthanides were detected by the circular dichroism positive band. The tetracycline complexes with the lanthanides of the second half of the yttrium subgroup (Ho(3+)-Lu3+) were characterized by the presence of the circular dichroism minimum at lambda(min)-425 nm. When the tetracycline concentration was above 1.5 x 10(-3) M, multiligand complexes with circular dichroism negative extremum at lambda(min)-400 nm formed.