Effects of lincomycin hydrochloride on the neurotoxicity of zebrafish.

Lincomycin hydrochloride is one of the commonly used drugs in clinic. However, it has many side effects on patients, and its mechanism is still poorly understood. In this study, 6 h post-fertilization (6 hpf) zebrafish embryos were exposed to several concentrations of lincomycin hydrochloride (15, 30, 60 µg/mL) for up to 24 or 96 hpf to detect their developmental toxicity and neurotoxicity, and to 6 days post-fertilization (6 dpf) to detect their behavioral toxicity. Our results showed that lincomycin hydrochloride could lead to embryonic head deformities (unclear ventricles, smaller ventricles, fewer new neurons). The studies showed that the frequency of spontaneous tail flick of zebrafish embryo increased at 24 hpf, and the lincomycin hydrochloride exposed zebrafish embryos showed increased heart rate, shorter body length, and yolk sac edema with severe pericardial edema at 96 hpf. The studies also showed that lincomycin hydrochloride increased oxidative stress level, Acetylcholinesterase (AChE) activity, ATPase activity and apoptosis in zebrafish larvae. In addition, the swimming behavior of zebrafish larvae decreased with the increase of lincomycin hydrochloride concentration, but the angular velocity and meandering degree increased, which might be due to the decreased activity of AChE and ATPase, as well as the decreased expression of genes related to neurodevelopment and neurotransmitter system, leading to the change of their motor behaviors. In summary, we found that lincomycin hydrochloride induced developmental toxicity and neurotoxicity in zebrafish larvae, contributing to a more comprehensive evaluation of the safety of the drug.

Dosage effects of lincomycin mycelial residues on lincomycin resistance genes and soil microbial communities.

Lincomycin mycelial residues (LMRs) are one kind of byproduct of the pharmaceutical industry. Hydrothermal treatment has been used to dispose of them and land application is an attractive way to reuse the treated LMRs. However, the safe dose for soil amendment remains unclear. In this study, a lab-scale incubation experiment was conducted to investigate the influence of the amendment dosage on lincomycin resistance genes and soil bacterial communities via quantitative PCR and 16S rRNA sequencing. The results showed that introduced lincomycin degraded quickly in soil and became undetectable after 50 days. Degradation rate of the high amendment amount (100 mg kg-1) was almost 4 times faster than that of low amendment amount (10 mg kg-1). Moreover, the introduced LMRs induced the increase of lincomycin resistance genes after incubation for 8 days, and two genes (lmrA and lnuB) showed a dosage-related increase. For example, the abundance of gene lmrA was 17.78, 74.13 and 128.82 copies g-1 soil for lincomycin concentration of 10, 50 and 100 mg kg-1, respectively. However, the abundance of lincomycin resistance genes recovered to the control level as the incubation period extended to 50 days, indicating a low persistence in soil. In addition, LMRs application markedly shifted the bacterial composition and significant difference was found between control soil, 10 mg kg-1 and 50 mg kg-1 lincomycin amended soil. Actually, several genera bacteria were significantly related to the elevation of lincomycin resistance genes. These results provided a comprehensive understanding of the effects of lincomycin dosage on the fate of resistance genes and microbial communities in LMRs applied soil.

Environmental fate and microbial effects of monensin, lincomycin, and sulfamethazine residues in soil.

The impact of commonly-used livestock antibiotics on soil nitrogen transformations under varying redox conditions is largely unknown. Soil column incubations were conducted using three livestock antibiotics (monensin, lincomycin and sulfamethazine) to better understand the fate of the antibiotics, their effect on nitrogen transformation, and their impact on soil microbial communities under aerobic, anoxic, and denitrifying conditions. While monensin was not recovered in the effluent, lincomycin and sulfamethazine concentrations decreased slightly during transport through the columns. Sorption, and to a limited extent degradation, are likely to be the primary processes leading to antibiotic attenuation during leaching. Antibiotics also affected microbial respiration and clearly impacted nitrogen transformation. The occurrence of the three antibiotics as a mixture, as well as the occurrence of lincomycin alone affected, by inhibiting any nitrite reduction, the denitrification process. Discontinuing antibiotics additions restored microbial denitrification. Metagenomic analysis indicated that Proteobacteria, Bacteroidetes, Actinobacteria, and Chloroflexi were the predominant phyla observed throughout the study. Results suggested that episodic occurrence of antibiotics led to a temporal change in microbial community composition in the upper portion of the columns while only transient changes occurred in the lower portion. Thus, the occurrence of high concentrations of veterinary antibiotic residues could impact nitrogen cycling in soils receiving wastewater runoff or manure applications with potential longer-term microbial community changes possible at higher antibiotic concentrations.

Uptake and effects of a mixture of widely used therapeutic drugs in Eruca sativa L. and Zea mays L. plants.

Pharmaceutically active compounds (PACs) are continuously dispersed into the environment due to human and veterinary use, giving rise to their potential accumulation in edible plants. In this study, Eruca sativa L. and Zea mays L. were selected to determine the potential uptake and accumulation of eight different PACs (Salbutamol, Atenolol, Lincomycin, Cyclophosphamide, Carbamazepine, Bezafibrate, Ofloxacin and Ranitidine) designed for human use. To mimic environmental conditions, the plants were grown in pots and irrigated with water spiked with a mixture of PACs at concentrations found in Italian wastewaters and rivers. Moreover, 10× and 100× concentrations of these pharmaceuticals were also tested. The presence of the pharmaceuticals was tested in the edible parts of the plants, namely leaves for E. sativa and grains for Z. mays. Quantification was performed by liquid chromatography mass spectroscopy (LC/MS/MS). In the grains of 100× treated Z. mays, only atenolol, lincomycin and carbamazepine were above the limit of detection (LOD). At the same concentration in E. sativa plants the uptake of all PACs was >LOD. Lincomycin and oflaxacin were above the limit of quantitation in all conditions tested in E. sativa. The results suggest that uptake of some pharmaceuticals from the soil may indeed be a potential transport route to plants and that these environmental pollutants can reach different edible parts of the selected crops. Measurements of the concentrations of these pharmaceuticals in plant materials were used to model potential adult human exposure to these compounds. The results indicate that under the current experimental conditions, crops exposed to the selected pharmaceutical mixture would not have any negative effects on human health. Moreover, no significant differences in the growth of E. sativa or Z. mays plants irrigated with PAC-spiked vs. non-spiked water were observed.

Toxic effect of the combined antibiotics ciprofloxacin, lincomycin, and tylosin on two species of marine diatoms.

The role that antibiotics and other "emerging contaminants" play in shaping environmental microbial communities is of growing interest. The use of the prokaryotic metabolic inhibitors tylosin (T), lincomycin (L), and ciprofloxacin (C) in livestock and humans is both global and extensive. Each of these antibiotic compounds exhibits an affinity for sediment particles, increasing the likelihood of their deposition in the benthos of aquatic systems and each are often present in environmental samples. The purpose of this study was to determine if T, L, and C and their mixtures exhibit significant toxicity to two species of marine diatoms, an algal class comprised of ubiquitous eukaryotic primary producers. Subpopulations from laboratory cultures of Cylindrotheca closterium and Navicula ramosissima were reared in 24-well microtiter plates in the presence of single or combined antibiotics in dilution series. Population growth rates were assessed via epifluorescent microscopic cell counts, from which the half-max inhibitory concentrations (IC(50)) were calculated and used as part of a toxic unit (TU) method for assessing mixture interactions. The single-compound IC(50)'s were, for C. closterium: T = 0.27 mg L(-1), L = 14.16 mg L(-1), C = 55.43 mg L(-1), and for N. ramosissima: T = 0.99 mg L(-1), L = 11.08 mg L(-1), C = 72.12 mg L(-1). These values were generally higher than similar metrics for freshwater species. Mixture IC(50)'s were generally synergistic against C. closterium and additive for N. ramosissima. Both single and combined treatments reduced or eliminated diatom motility. Monochemical responses were similar between species and were not useful for predicting mixture interactions. Mixtures had compound-specific and species-specific effects, favoring N. ramosissima. These results suggest that anthropogenic antibiotics may play a significant role in the ecology of environmental benthic microbial communities. They also suggest single-compound/species studies do not yield useful predictions of the ecological impact of anthropogenic pharmaceuticals.

Gene expression of ribosomal protein mRNA in Chironomus riparius: effects of endocrine disruptor chemicals and antibiotics.

Ribosomal protein genes are essential for cellular development. To examine the effects of ribosomal protein genes under various cellular stress conditions in chironomids, ribosomal protein S3 (RpS3) and S6 (RpS6) cDNA from Chironomus riparius were characterized and their expression was analyzed during development. A comparative and phylogenetic study among different orders of insects was carried out by analysis of sequence databases. C. riparius RpS3 was highly conserved at the protein level and shared over 85% amino acid identity with homologous sequences from other insects. RpS6 also showed approximately 80% amino acid identity. The RpS3 and S6 transcripts were present during different developmental stages but were most abundant during the embryonic stage. Furthermore, expression of the previously reported ribosomal proteins RpL11, L13, and L15, as well as RpS3 and S6 was analyzed following exposure to various concentrations of three endocrine disruptor chemicals (EDCs), di(2-ethylhexyl) phthalate, bisphenol A, and 4-nonylphenol (4NP), and the veterinary antibiotics (VAs) fenbendazole, sulfathiazole, and lincomycin. Only RpS3 gene expression was up-regulated significantly in response to EDCs and fenbendazole. However, the C. riparius ribosomal proteins showed a limited response to cellular stress, following exposure to EDCs and VAs.

Effect of chronic exposure to acetaminophen and lincomycin on Japanese medaka (Oryzias latipes) and freshwater cladocerans Daphnia magna and Moina macrocopa, and potential mechanisms of endocrine disruption.

Chronic toxicity of acetaminophen and lincomycin were evaluated using freshwater organisms including two crustaceans (Daphnia magna and Moina macrocopa) and a fish (Oryzias latipes). H295R, a human adrenal cell was also used to understand the effects on steroidogenesis. In 21 d D. magna exposure, survival NOEC was found at 5.72 mg L(-1) and no reproduction related effects were noted at this level of exposure to acetaminophen, while 21 d survival or growth effects were not observed even at the highest exposure levels (153 mg L(-1)) for lincomycin. In the chronic fish toxicity test, significant reduction in juvenile survival was observed at 30 d post-hatch (dph) at 95 mg L(-1) of acetaminophen, and 0.42 mg L(-1) of lincomycin. After the exposure to both pharmaceuticals, vitellogenin levels tended to increase in male fish at 90 dph. In the eggs which were prenatally exposed to 9.5 mg L(-1) of acetaminophen, reduced hatchability was observed. The results of H295R cell assay showed that both pharmaceuticals could alter steroidogenic pathway and increase estrogenicity. Endocrine disruption potentials and their ecological implication may deserve further studies. Our observations suggest however that ecological risks of both pharmaceuticals are negligible at the concentrations currently found in the environment.

Micronucleus test and comet assay for the evaluation of zebrafish genomic damage induced by erythromycin and lincomycin.

An enormous quantity of pharmacologically active principles are currently being introduced into the environment, with consequent escalation of environmental problems, but only a small number of studies are focusing on an assessment of their genotoxic effects. The aim of this article is to assess the genotoxic effects of erythromycin, lincomycin, and of a combination of these two antibiotics on the genome of the zebrafish. The genotoxicity of the two antibiotics was assessed by applying the micronucleus test to erythrocytes and performing a Comet assay on erythrocytes and hepatocytes. The fish were exposed to antibiotics at different concentrations and times of exposure, under standard laboratory conditions. Depending on the different experimental conditions, erythromycin and lincomycin induced a significant increase in DNA migration (tail moment) and a significant increase in micronuleus frequency. We also conducted an analysis on the activation of repair mechanisms when the genotoxic agent was removed. Only a few of the cells displayed a decrease in damage under these test conditions.

Effects and interactions in an environmentally relevant mixture of pharmaceuticals.

With the goal of assessing the environmental risk of pharmaceuticals, we have previously observed that a mixture of 13 different drugs at environmentally relevant concentrations had adverse consequences on human and zebra fish cells in vitro. Here we aimed to identify both main and interaction effects within the same environmentally relevant mixture of pharmaceuticals. We studied in vitro cytotoxicity in Escherichia coli, human embryonic HEK293, and estrogen-responsive OVCAR3 tumor cells using fractional-factorial experimental design. Our approach identified a subset of compounds of primary environmental concern, namely atenolol, bezafibrate, ciprofloxacin, and lincomycin, that had statistically significant effects on prokaryotic and eukaryotic cells at environmentally relevant exposure levels (ng/l). Drugs could interact and behave as chemosensitizers, with joint effects representing a statistically significant element of mixture toxicity. Effects and interactions were concentration dependent, confirming the difficulty of dose extrapolation in mixture toxicity data. This study suggests that a thorough investigation of mixture effects can direct environmental concerns toward a handful of pharmaceuticals, which may represent an actual risk at environmental concentrations. We indicate that risk identification may strongly depend on the use of environmentally relevant exposure scenarios. Antagonistic-synergistic interactions and dose dependency of effects may hamper the modeling and prediction of mixture toxicity with pharmaceuticals. Hazard identification for micropollutants depends heavily on appropriate study designs, and we indicate the use of in vitro cytotoxicity threshold and statistical design of experiments (DOEs) as a valid approach.

Biodegradability and toxicity of pharmaceuticals in biological wastewater treatment plants.

In this experimental study both biological treatability of pharmaceuticals and their potential toxic effect in biological processes were evaluated. The pharmaceuticals were selected among those that are present at higher concentration in the Italian wastewater treatment plant effluents and widely used as antiulcer (ranitidine), beta-blocker (atenolol) and antibiotic (lincomycin). The present paper is the continuation of a work already presented,[1] which used a synthetic wastewater fed to laboratory scale SBR (Sequencing Batch Reactor) operated with different sludge ages (8 and 14 days), different biochemical conditions (aerobic or anoxic-aerobic mode) and several influent drug concentrations (2, 3 and 5 mg/L). In this case a real municipal wastewater was used as influent to the SBR. In parallel, batch tests were conducted to determine the removal kinetics of drugs and nitrogen. Toxicity tests using a titrimetric biosensor to verify possible inhibition on microorganisms were also performed. Finally, the possible adsorption of the pharmaceuticals on activated sludge was evaluated. The drugs under investigation showed different behaviours in terms of both biodegradability and toxicity effect on nitrifiers. Ranitidine showed generally low removal efficiencies (17-26%) and a chronic inhibition on nitrification. Atenolol showed generally higher removal efficiencies than ranitidine, even if the fairly good efficiency obtained in the previous experimentation with synthetic wastewater (up to 90%) was not attained with real wastewater (36%). No inhibition on nitrification was observed on both acclimated and non acclimated microorganisms with a high nitrification activity, whilst it was present with activated sludge characterised by a lower nitrification activity. Consistently with his pharmaceutical properties, lincomycin showed significant inhibition on nitrification activity.

Lincomycin solar photodegradation, algal toxicity and removal from wastewaters by means of ozonation.

Antibiotic molecules have been reported among the xenobiotics present at trace levels in sewage treatment plant (STP) effluents and aquatic environment. Lincomycin, one of the most used in clinical practices whose presence in the STP effluents has been often documented, is submitted to an extensive investigation to assess its persistence in the environment and toxicity towards different algal strains. The possibility to remove the lincomycin from water by means of ozonation is demonstrated and a reduction of toxicity of ozonated solutions on S. leopoliensis, with respect to untreated solutions containing this compound, is obtained even just for 1h of treatment. Kinetic constants for the attack to lincomycin of ozone (from 1.53 x 10(5) M(-1)s(-1) at pH = 3.0 and 4.93 x 10(5) M(-1)s(-1) at pH = 6.7) and OH radicals (4.37 x 10(9) M(-1)s(-1) at pH = 5.5 and 4.59 x 10(9) M(-1)s(-1) at pH = 7.5) are also evaluated.

Macrolides and lincomycin susceptibility of Mycoplasma hyorhinis and variable mutation of domain II and V in 23S ribosomal RNA.

A total of 151 strains of Mycoplasma hyorhinis isolated from porcine lung lesions (weaned pigs, n=71, and finishers, n=80) were investigated for their in vitro susceptibility to 10 antimicrobial agents. Thirty-one strains (28 from weaned pigs and 3 from finishers) showed resistance to 16-membered macrolide antibiotics and lincomycin. The prevalence of the 16-membered macrolide-resistant M. hyorhinis strain in weaned pigs from Japanese herds has approximately quadrupled in the past 10 years. Several of the 31 strains were examined for mutations in the 23S ribosomal RNA (rRNA). All field strains tested showed a transition of A to G at position 2059 of 23S rRNA-rendered Escherichia coli. On the other hand, individual tylosin- and lincomycin-resistant mutants of M. hyorhinis were selected in vitro from the susceptible type strain BTS7 by 3 to 9 serial passages in subinhibitory concentrations of each antibiotic. The 23S rRNA sequences of both tylosin and lincomycin-resistant mutants were compared with that of the radical BTS7 strain. The BTS7 mutant strain selected by tylosin showed the same transition as the field-isolated strains of A2059G. However, the transition selected in lincomycin showed mutations in domains II and V of 23S rRNA, G2597U, C2611U in domain V, and the addition of an adenine at the pentameric adenine loop in domain II. The strain selected by lincomycin showed an additional point mutation of A2062G selected by tylosin.

Antibiotic sensitivity patterns among Indian strains of avian Pasteurella multocida.

An investigation was carried out to study the antibiotic sensitivity of avian strains of Pasteurella multocida and to select an effective antimicrobial agent for control of avian pasteurellosis in India. A total of 123 strains of P. multocida recently isolated from different avian species (chicken, duck, turkey, quail, and goose), from different regions of India were subjected to antibiotic sensitivity tests using 20 different antibiotics. Absolute resistance was observed against sulfadiazine. The studies indicated that the strains were most sensitive to chloramphenicol (73.98%), followed by enrofloxacin (71.54%), lincomycin (64.23%), norfloxacin (61.79%) and doxycycline-HCl (56.91%). The majority of the strains were found to exhibit intermediate sensitivity. Chloramphenicol was selected and suggested for treatment. Antibiogram studies also revealed the emergence of multidrug-resistant strains of P. multocida among Indian poultry.

Antibiotic susceptibility of Mycoplasma hyopneumoniae isolated from swine.

The antibiotic susceptibility of thirty-nine strains of Mycoplasma hyopneumoniae isolated from swine between 1970-1981 and 1989-1990 was investigated. From the present results, it is suggested that the susceptibility to chlortetracycline has been decreasing in Japan. On the other hand, all the strains were sensitive to lincomycin, thiamphenicol and macrolides. Newly developed macrolides such as tilmicosin, acetyl-isovaleryl-tylosin and mirosamycin had equal or higher activity than general macrolides.

[Substantiation of hygienic standard of lincomycin in the air of the work area]. / Obosnovanie gigienicheskogo standarta linkomitsina v vozdukhe rabochei zony.

The main parameters of lincomycin toxicometry were studied in animals. It was shown that the compound was low toxic after its oral or intraperitoneal administration in single doses, had no local irritant and skin resorptive effects and did not accumulate. The allergenic properties were slightly pronounced. The intoxication picture after a single inhalation was characterized by renal dysfunction, erythropenia, neutrophilia, lymphopenia and impairment of the normal intestinal microflora. The zone of the specific antimicrobial effect was equal to 8. On chronic inhalation, the signs of the specific antimicrobial effect were of the paramount importance: Limch am was equal to 4.7 mg/m3 and Limch exceeded 18.3 mg/m3. In the concentrations used, the substance had no embryotoxic and gonadotropic effects. The level of 0.5 mg/m3 (for Hazard Class 2) was recommended and approved as the maximum allowable concentration.

[Toxicological and pharmacokinetic research on lincomycin hydrochloride in broiler chickens and layer hens]. / Toksikologichni i farmakokinetichni prouchvaniia na linkomitsinov khidrokhlorid pri pileta broileri i kokoshki.

Tested was the activity of linkomycin hydrochloride at the rate of 820 IU/mg. The LD50 of the preparation was found to exceed 12.0 g/kg body mass at intracrop application to broiler birds. When offered with the drinking water or the feed in amounts that were 3 to 5 times higher than the therapeutic ones for broilers in the course of 45 days it did not cause mortality nor induced changes in the white and red blood cell counts, in the total protein, and in the levels of urea, blood sugar, AlT, and AcT. Neither were there any structural changes in the internal organs. Upon the single intracrop treatment of broilers with linkomycin at 50 and 100 mg/kg most of the amount introduced was established in the blood serum at the second hour and up to the 6th and 8th hour, respectively. Studies on the effect of age on the serum concentrations at this route of introduction revealed that the preparation was absorbed at higher rates in the broilers than in the layers, while with the persistence of the levels there were no essential differences. At the single oral administration with broilers in amounts of 100 mg/kg linkomycin was chiefly retained in the kidneys, liver, and lungs as well in the content of the small and large intestines. At intracrop application to layers in doses of 100 mg/kg in the course of 7 days the antibiotic was excreted with the eggs at the time of treatment and in the course of 4 days later in bacteriostatic concentrations of 0.6-5.9 micrograms/cm3.

[Embryotoxic and teratogenic effects of Pharmachim's lincomycin hydrochloride]. / Embriotoksichen i teratogenen efekt na linkomitsinov khidrokhlorid "Farmakhim".

Tested was the teratogenic and embryotoxic action of linkomycin hydrochloride with both placental and nonplacental animals. It was found that when applied twice, orally, to albino rats on the 4th day of pregnancy at the rate of 250 mg/kg body mass (=5 ED50), and once to chick embryos at 50 mg per egg the antibiotic raised the percent of pre- and postimplantation loss of embryos. In a series of applications (oral) from the 3rd up to the 19th day at 50 mg/kg body mass to rats as well as in a single application to chick embryos at 10, 20, and 50 mg per egg linkomycin showed no teratogenic effects.

[Experimental study of lincomycin ointment and gel]. / Eksperimental'noe izuchenie mazi i gelia linkomitsina.

The pharmacokinetics and safety of the lincomycin ointment and gel were studied. It was shown that diffusion of lincomycin through the skin was satisfactory. Investigation of their general toxicity and organotropic properties revealed neither irritating effect nor changes in the internal organs associated with the toxic effect of the drugs. On the basis of the data on the stability of the lincomycin ointment and gel obtained on their storage the lincomycin ointment was recommended for industrial production.

Effects of lincomycin and tetracycline on production and properties of enterotoxins of enterotoxigenic Escherichia coli.

Enterotoxigenic Escherichia coli grown in the presence of lincomycin and tetracycline produced an increased amount of heat-labile enterotoxin (LT). These antibiotics increased the production of not only extracellular LT but also intracellular LT. On the other hand, lincomycin did not stimulate the production of heat-stable enterotoxin by enterotoxigenic E. coli. The extracellular LTs produced in the presence of lincomycin and tetracycline were purified and analyzed by sodium dodecyl sulfate-polyacrylamide gel disc electrophoresis. Results showed that the A subunits of the purified LTs were not nicked, unlike that of extracellular LT produced in the absence of the antibiotics.