Degradation of sulfadiazine, sulfachloropyridazine and sulfamethazine in aqueous media.

Antibiotics discharged to the environment constitute a main concern for which different treatment alternatives are being studied, some of them based on antibiotics removal or inactivation using by-products with adsorbent capacity, or which can act as catalyst for photo-degradation. But a preliminary step is to determine the general characteristics and magnitude of the degradation process effectively acting on antibiotics. A specific case is that of sulfonamides (SAs), one of the antibiotic groups most widely used in veterinary medicine, and which are considered the most mobile antibiotics, causing that they are frequently detected in both surface- and ground-waters, facilitating their entry in the food chain and causing public health hazards. In this work we investigated abiotic and biotic degradation of three sulfonamides (sulfadiazine -SDZ-, sulfachloropyridazine -SCP-, and sulfamethazine -SMT-) in aqueous media. The results indicated that, in filtered milliQ water and under simulated sunlight, the degradation sequence was: SCP > SDZ ≈ SMT. Furthermore, the rate of degradation clearly increased with the raise of pH: at pH 4.0, half-lives were 1.2, 70.5 and 84.4 h for SCP, SDZ and SMT, respectively, while at pH 7.2 they were 2.3, 9.4 and 13.2 h for SCP, SMT and SDZ. The addition of a culture medium hardly caused any change in degradation rates as compared to experiments performed in milliQ water at the same pH value (7.2), suggesting that in this case sulfonamides degradation rate was not affected by the presence of some chemical elements and compounds, such as sodium, chloride and phosphate. However, the addition of bacterial suspensions extracted from a soil and from poultry manure increased the rate of degradation of these antibiotics. This increase in degradation cannot be attributed to biodegradation, since there was no degradation in the dark during the time of the experiment (72 h). This indicates that photo-degradation constitutes the main removal mechanism for SAs in aqueous media, a mechanism that in this case was favored by humic acids supplied with the extracts from soil and manure. The overall results could contribute to the understanding of the environmental fate of the three sulfonamides studied, aiding to program actions that could favor their inactivation, which is especially relevant since its dissemination can involve serious environmental and public health risks.

Tissue residue distribution and withdrawal time estimation of trimethoprim and sulfachloropyridazine in Yugan black-bone fowl (Gallus gallus domesticus Brisson).

Black-bone fowl are different from ordinary broilers in appearance and are considered to have rich nutritional properties. However, the metabolism of therapeutic drugs in black-bone fowl remains unclear. This study aimed to determine the tissue residue depletion kinetics of trimethoprim and sulfachloropyridazine in Yugan black-bone fowl, after daily oral administrations for 5 days at 4 mg/kg bw/day trimethoprim and 20 mg/kg bw/day sulfachloropyridazine, and to calculate the withdrawal times. After consecutive oral administrations, the tissues (liver, kidney, muscle and skin/fat) were collected at each of the following time points (0.16, 1, 3, 5, 7, 9, 20, 30 and 40 days). A newly-devised LC-MS/MS method was used to analyse the concentrations of trimethoprim and sulfachlorpyridazine in target tissues. The results showed that sulfachloropyridazine was rapidly metabolised in broilers, and there was no residue in all tissues 3 days post-administration. The concentration of trimethoprim in black-bone fowl skin/fat is the highest, and its metabolism rate is low. After 40 days, the concentration of trimethoprim in skin/fat is still as high as 140.1 ± 58.0 µg/kg, exceeding the maximum residue limit. In order to protect consumers' health, it is suggested that the withdrawal time of TMP in Yugan black-bone fowl is 69 days.

Studies on removing sulfachloropyridazine from groundwater with microbial bioreactors.

Sulfachloropyridazine (SCP), an antibiotic used in aquaculture and in animal husbandry, is a common contaminant in surface and groundwaters. Two types of microbial reactors were evaluated as methods for removing SCP from flowing water. One type of reactor evaluated was a nitrogen-limiting biobarrier; the other a slow-sand-filter. Results showed that the soybean oil-fed, nitrogen-limiting biobarrier was not very effective at removing SCP from flowing water. When supplied with flowing water containing 2.4 mg l(-1) SCP the nitrogen-limiting biobarrier removed ~0.6 mg l(-1) SCP or about 28% of that present. SCP removal by the nitrogen-limiting biobarrier may not have been biological as abiotic removal was not ruled out. More efficient biological removal was obtained with the slow-sand-filter which reduced the SCP levels from 2.35 to 0.048 mg l(-1), a removal efficiency of ~98%. High levels of nitrate nitrogen, 50 mg l(-1) N, did not interfere with the removal processes of either reactor suggesting that SCP was not being degraded as a microbial nitrogen source.

Effects of sulfachlorpyridazine in MS.3-arable land: a multispecies soil system for assessing the environmental fate and effects of veterinary medicines.

A multispecies soil system (MS.3) has been used to evaluate the ecological effects of veterinary pharmaceuticals in soil as a result of routine agricultural practices. Different experimental conditions were tested and the variation of the different parameters was evaluated for a final design. A protocol for the MS.3-arable land is presented here. Emergence of seedlings, plant elongation and biomass, earthworm mortality, and soil microbial enzymatic activities have been selected as toxicological endpoints for soil organisms. Toxicity tests were conducted with the leachate on aquatic organisms (in vitro fish cell lines, daphnids, and algae). The system was used for assessing the effects of the antimicrobial sulfachlorpyridazine that was tested in triplicate at concentrations of 0.01, 1, and 100 mg/kg. The chemical was mixed uniformly with a 20-cm depth soil column to resemble the distribution of manure within arable soil. Reversible and nonreversible effects on soil enzymatic activities were observed at 1 and 100 mg/kg, respectively. Earthworms were not affected. Significant reduction of plant elongation and biomass was observed at the highest concentration. Degradation and leaching contributed to the dissipation of sulfachlorpyridazine from the soil column. The undiluted leachate was highly toxic to Daphnia magna. The parent chemical was assumed responsible for the leachate toxicity although the role of mobile metabolites could not be excluded fully. No significant effects were observed for green algae Chlorella vulgaris and for the rainbow trout established cell lines RTG-2 (rainbow trout gonads) and RTL-WI (rainbow trout liver). The MS.3 system offers a cost-effective experimental approach to measure simultaneously fate and effects of chemicals on a realistic soil system under controlled laboratory conditions. The advantages of using MS.3-effect endpoints are discussed.

The sorption and transport of a sulphonamide antibiotic in soil systems.

Veterinary medicines are administered to animals to treat disease and protect their health. After administration, the substances can be metabolised and a mixture of the parent compound and metabolites may be excreted in the urine and faeces. For animals on pasture, the excreta will be released directly to soil whereas for intensively reared animals, the main route of entry will be through slurry and manure spreading. Whilst the behaviour of other classes of substance (e.g. pesticides and nutrients) that are applied to soil is well understood, limited information is available on the transport and fate of veterinary medicines applied to soils. Laboratory and field studies were, therefore, performed to investigate the sorption behaviour of the sulfonamide antibiotic, sulfachloropyridazine, in soil and to assess the potential for sulfachloropyridazine to move from soil to surface waters and groundwaters. Sorption coefficients (K(D)) for the compound in soil and soil/slurry mixtures were low (ranging from 0.9 to 1.8 l kg(-1)) and indicated that the substance would be highly mobile. Field studies on a clay field supported these observations and demonstrated that, after application, the compound was rapidly transported to surface waters, concentrations of up to 590 microg l(-1) being observed in drainage waters. Leaching studies at a sandy site indicated that the substance had a low potential to leach to groundwaters, concentrations in the soil pore water being below or close to analytical detection limits. An assessment of currently available models for predicting concentrations of veterinary medicines entering surface waters indicated that for sulfachloropyridazine, the methods provide reasonable estimates, predicted concentrations being within a factor of two of the maximum measured concentrations. The approaches may not, however, be appropriate for use on highly hydrophobic substances or for predicting groundwater concentrations.

Fate of veterinary antibiotics in a macroporous tile drained clay soil.

The environment may be exposed to veterinary medicines administered to livestock through the application of organic fertilizers to land. For other groups of substances that are applied to agricultural land (e.g., pesticides), preferential flow in underdrained clay soils has been identified as an extremely important mechanism by which pollution of surface waters can occur. This study, therefore, was performed to investigate the fate of three antibiotics from the sulfonamide, tetracycline, and macrolide groups. Pig slurry was applied to a field in arable production in two consecutive years and the fate of the compounds was monitored in the soil and drainage water. Both sulfachloropyridazine and oxytetracycline were detected in soil at concentrations up to 365 and 1691 microg/kg, respectively. Subsequently, peak concentrations of the two substances in drainflow were 613.2 and 36.1 microg/L, although mass losses to the receiving water were less than 0.5%. In contrast, tylosin was not detected at all. These findings could be explained by the persistence and sorption characteristics of the antibiotics, while preferential flow via desiccation cracks and worm channels to the tile drains was found to be the most important route for translocation of the chemicals. Thus, when the soil was disced prior to slurry application, losses were reduced significantly. It is evident that processes governing pesticide fate also apply to veterinary antibiotics.

Influence of hepatic and renal disorders on the pharmacokinetics of sulfachloropyridacine.

The pharmacokinetics of sulfachloropyridacine were studied in a series of 30 adult patients (healthy volunteers, patients with moderate hepatic impairment and patients with renal impairment). In all cases a 500 mg dose of sodium sulfachloropyridacine was administered orally. The drug follows a single compartment pharmacokinetic model. In healthy patients the following pharmacokinetic parameters were established: Ka = 5.130 h-1, Ke = 0.205 h-1, tmax = 40 min, Vd = 7.94 liters, and in patients diagnosed with cirrhosis a decrease is appreciable in the absorption constant and in the area below the blood-time levels curve. A linear relationship is established between the elimination constant and creatinine clearance. A dosage regimen, applicable to patients with renal impairment, is established as a function of the pharmacokinetic parameters. The degree of plasma protein binding of sulfachloropyridacine diminishes significantly in patients with renal impairment.

Pharmacokinetic analysis of the level of sulfonamide-trimethoprim combination in calves.

Pharmacokinetic parameters which describe biological availability (AUC, Cmax, tmax) and elimination (kel, t 1/2) of sulfonamides and trimethoprim were determined in calves. Pharmacokinetic analysis was based upon "one compartment model". The average +/- SD half-life in plasma was for sulfametoxazole 12.8 +/- 0.32 h, for sulfamerazine 17.0 +/- 0.75 h and for sulfachloropyridazine 13.1 +/- 0.86 h. The average half-life of trimethoprim in plasma was 11.3 +/- 0.56 h. In fasted calves the concentrations of sulfachloropyridazine and trimethoprim in plasma were higher as compared with fed group.

Postnatal development of renal function in piglets: changes in excretory pattern of sulphachlorpyridazine.

The renal excretion of sulphachlorpyridazine (SCP) was measured in 34 unanaesthetized piglets 5-74 days of age. The renal handling of SCP was found to include: glomerular filtration, active tubular secretion and passive reabsorption. During the age period studied the clearance of ultrafiltrable SCP increased several times more than GFR (inulin clearance) indicating that the balance between filtration, secretion and reabsorption is changed during postnatal maturation. The age-related change in clearance ratio possibly reflects both a greater increase in tubular secretory capacity than in glomerular filtration and a relative decrease in the passive reabsorption.

In vitro and in vivo binding of trimethoprim and sulphachlorpyridazine to equine food and digesta and their stability in caecal contents.

Binding of antibiotics to food has received little attention in equine medicine, although such binding could potentially reduce the bioavailability and clinical efficacy. In the present study, binding of trimethoprim (TMP) and sulphachlorpyridazine (SCP) to hay, grass silage and concentrate was investigated in vitro in buffer at pH 6.8 at different concentrations. The binding of TMP and SCP to caecal contents was also studied. In addition, the degradation of TMP and SCP by the caecal microflora was investigated by incubating sterilized and non-sterilized caecal contents for 3 h at 37 degrees C under anaerobic conditions and comparing the TMP and SCP contents. Further, a TMP/SCP powder formulation was adminstered orally with concentrate at a dose rate of 5 mg/kg TMP and 25 mg/kg SCP to three ponies with a caecum fistula; the animals were deprived of food for 8 h before administration. Blood samples, caecal contents samples and faecal samples were collected and analysed for TMP and SCP concentrations by means of high performance liquid chromatography (HPLC). Three non-fistulated ponies, acting as control animals, were fed the same dose of TMP/SCP with concentrate after 8 h of food deprivation and blood samples were taken. The percentage of in vitro binding of TMP as well as SCP to hay, grass silage and concentrate at concentrations of 4 micrograms/mL to 10 micrograms/mL was high (60-90%). TMP and SCP were also extensively bound to caecal contents (50-70%). At spiking concentrations above 10 micrograms/mL the percentage of binding decreased. There was no evidence of biodegradation of TMP or SCP in caecal contents. In vivo, both drugs could be detected in the caecal contents and in the faeces of three fistulated ponies. However, the fistulated ponies differed from the control ponies in that their TMP and SCP plasma concentrations were higher, and two fistulated ponies did not show double peaks in their plasma concentration-time curves. Therefore, the fistulated ponies did not provide an optimal model for in vivo binding studies. Despite this limitation, it can be concluded that binding of TMP and SCP to food is a major cause of the limited bioavailability of these drugs in the horse. It is hypothesized that the binding is reversible, and that a second absorption phase occurs in the large intestine, but part of the administered dose remains bound as both drugs were found in the faeces.