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Background: The TetR family of transcriptional regulators (TFRs), serving as crucial regulators of diverse cellular processes, undergo conformational changes induced by small-molecule ligands, which either inhibit or activate them to modulate target gene expression. Some ligands of TFRs in actinomycetes and their regulatory effects have been identified and studied; however, regulatory mechanisms of the TetR family in the lincomycin-producing Streptomyces lincolnensis remain poorly understood. Results: In this study, we found that AbrT (SLCG_1979), a TetR family regulator, plays a pivotal role in regulating lincomycin production and morphological development in S. lincolnensis. Deletion of abrT gene resulted in increased lincomycin A (Lin-A) production, but delayed mycelium formation and sporulation on solid media. AbrT directly or indirectly repressed the expression of lincomycin biosynthetic (lin) cluster genes and activated that of the morphological developmental genes amfC, whiB, and ftsZ. We demonstrated that AbrT bound to two motifs (5′-CGCGTACTCGTA-3′ and 5′-CGTACGATAGCT-3′) present in the bidirectional promoter between abrT and SLCG_1980 genes. This consequently repressed abrT itself and its adjacent gene SLCG_1980 that encodes an arabinose efflux permease. D-arabinose, not naturally occurring as L-arabinose, was identified as the effector molecule of AbrT, reducing its binding affinity to abrT-SLCG_1980 intergenic region. Furthermore, based on functional analysis of the AbrT homologue in Saccharopolyspora erythraea, we inferred that the TetR family regulator AbrT may play an important role in regulating secondary metabolism in actinomycetes. Conclusions: AbrT functions as a regulator for governing lincomycin production and morphological development of S. lincolnensis. Our findings demonstrated that D-arabinose acts as a ligand of AbrT to mediate the regulation of lincomycin biosynthesis in S. lincolnensis. Our findings provide novel insights into ligand-mediated regulation in antibiotic biosynthesis. [ABSTRACT FROM AUTHOR]

SUMMARY: Enhancing the efficiency of photosynthesis represents a promising strategy to improve crop yields, with keeping the steady state of PSII being key to determining the photosynthetic performance. However, the mechanisms whereby the stability of PSII is maintained in oxygenic organisms remain to be explored. Here, we report that the Psb28 protein functions in regulating the homeostasis of PSII under different light conditions in Arabidopsis thaliana. The psb28 mutant is much smaller than the wild‐type plants under normal growth light, which is due to its significantly reduced PSII activity. Similar defects were seen under low light and became more pronounced under photoinhibitory light. Notably, the amounts of PSII core complexes and core subunits are specifically decreased in psb28, whereas the abundance of other representative components of photosynthetic complexes remains largely unaltered. Although the PSII activity of psb28 was severely reduced when subjected to high light, its recovery from photoinactivation was not affected. By contrast, the degradation of PSII core protein subunits is dramatically accelerated in the presence of lincomycin. These results indicate that psb28 is defective in the photoprotection of PSII, which is consistent with the observation that the overall NPQ is much lower in psb28 compared to the wild type. Moreover, the Psb28 protein is associated with PSII core complexes and interacts mainly with the CP47 subunit of PSII core. Taken together, these findings reveal an important role for Psb28 in the protection and stabilization of PSII core in response to changes in light environments. Significance Statement: The small protein Psb28 plays a crucial role in the protection and stabilization of PSII reaction center via direct interaction with the CP47 subunit of PSII core complexes in the model plant Arabidopsis thaliana, providing novel insights into the maintenance of PSII function for boosting photosynthetic capacity in response to changing light environments. [ABSTRACT FROM AUTHOR]

In this study, a liquid chromatographic method was developed for the fast determination of lincomycin, polymyxin and vancomycin in a preservation solution for transplants. A Kinetex EVO C18 (150 × 4.6 mm, 2.6 µm) column was utilized at 45 ◦C. Gradient elution was applied using a mixture of mobile phases A and B, both including 30 mM phosphate buffer at pH 2.0 and acetonitrile, at a ratio of 95:5 (v/v) for A and 50:50 (v/v) for B. A flow rate of 1.0 mL/min, an injection volume of 20 µL and UV detection at 210 nm were used. A degradation study treating the three antibiotics with 0.5 M hydrochloric acid, 0.5 M sodium hydroxide and 3% H2O2 indicated that the developed method was selective toward lincomycin, polymyxin, vancomycin and their degradation products. Other ingredients of the preservation solution, like those from the cell culture medium, did not interfere. The method was validated with good sensitivity, linearity, precision and accuracy. Furthermore, lincomycin, polymyxin and vancomycin were found to be stable in this preservation solution for 4 weeks when stored at −20 ◦C. [ABSTRACT FROM AUTHOR]

Background: Antibiotic residues come from food of animal origin, such as broiler chicken, have a variety of consequences on human health and increase the likelihood of antibiotic resistance. Lincomycin residue investigations in broiler chicken especially in plasma broiler chicken should be undertaken utilising the validation method analysis. Aim: The purpose of this study is to determine the high-performance liquid chromatography (HPLC) as a validation method for calculating the residual concentration of lincomycin in broiler chicken blood plasma and compare it with the Minimum Inhibitor Concentration (MIC) and Maximum Residue Limits (MRL) standards for lincomycin. Methods: Thirty-five-day-old broiler chickens cobb 700 were weighed and randomly allocated to and separated into control (placebo) and six treatment groups of varying doses and duration. The treatment group's suggested dosage of lincomycin was 50, 100, or 150 mg/kg/day given to 18-day-old chicken, along with drinking water for a week (A group) and two weeks (P group). Lincomycin levels in blood plasma were validated using HPLC. The residual lincomycin concentrations 24 hours and one week after injection were compared to the lincomycin MIC and the Indonesian National Standard of MRL. Result: The validation of lincomycin reveals a linear value in blood plasma with an R2 of 0.9983. Precision and accuracy levels indicate promising results for detecting lincomycin. The retention duration for 100 µg/mL lincomycin was 10.0-10.5 minutes. Lincomycin had LOD and LOQ values of 13.98 and 4.86 µg/mL, respectively. After one week of dosing at 50 and 100 mg/kg dosages, lincomycin residue detection was 0.00, which was below the MRL criterion of [Open Vet J 2024; 14(6.000): 1453-1459]

The lincoamide antibiotic lincomycin, derived from Streptomyces lincolnensis, is widely used for the treatment of infections caused by gram-positive bacteria. As a common global regulatory factor of GntR family, DasR usually exists as a regulatory factor that negatively regulates antibiotic synthesis in Streptomyces. However, the regulatory effect of DasR on lincomycin biosynthesis in S. lincolnensis has not been thoroughly investigated. The present study demonstrates that DasR functions as a positive regulator of lincomycin biosynthesis in S. lincolnensis, and its overexpression strain OdasR exhibits a remarkable 7.97-fold increase in lincomycin production compared to the wild-type strain. The effects of DasR overexpression could be attenuated by the addition of GlcNAc in the medium in S. lincolnensis. Combined with transcriptome sequencing and RT-qPCR results, it was found that most structural genes in GlcNAc metabolism and central carbon metabolism were up-regulated, but the lincomycin biosynthetic gene cluster (lmb) were down-regulated after dasR knock-out. However, DasR binding were detected with the DasR responsive elements (dre) of genes involved in GlcNAc metabolism pathway through electrophoretic mobility shift assay, while they were not observed in the lmb. These findings will provide novel insights for the genetic manipulation of S. lincolnensis to enhance lincomycin production. Key points: • DasR is a positive regulator that promotes lincomycin synthesis and does not affect spore production • DasR promotes lincomycin production through indirect regulation • DasR correlates with nutrient perception in S. lincolnensis [ABSTRACT FROM AUTHOR]

Key message: A GLK homologue was identified and functionally characterized in Catharanthus roseus. Silencing CrGLK with VIGS or the chloroplast retrograde signaling inducer lincomycin increased terpenoid indole alkaloid biosynthesis. Catharanthus roseus is the sole source of the chemotherapeutic terpenoid indole alkaloids (TIAs) vinblastine and vincristine. TIA pathway genes, particularly genes in the vindoline pathway, are expressed at higher levels in immature versus mature leaves, but the molecular mechanisms responsible for this developmental regulation are unknown. We investigated the role of GOLDEN2-LIKE (GLK) transcription factors in contributing to this ontogenetic regulation since GLKs are active in seedlings upon light exposure and in the leaf’s early development, but their activity is repressed as leaves age and senesce. We identified a GLK homologue in C. roseus and functionally characterized its role in regulating TIA biosynthesis, with a focus on the vindoline pathway, by transiently reducing its expression through two separate methods: virus-induced gene silencing (VIGS) and application of chloroplast retrograde signaling inducers, norflurazon and lincomycin. Reducing CrGLK levels with each method reduced chlorophyll accumulation and the expression of the light harvesting complex subunit (LHCB2.2), confirming its functional homology with GLKs in other plant species. In contrast, reducing CrGLK via VIGS or lincomycin increased TIA accumulation and TIA pathway gene expression, suggesting that CrGLK may repress TIA biosynthesis. However, norflurazon had no effect on TIA gene expression, indicating that reducing CrGLK alone is not sufficient to induce TIA biosynthesis. Future work is needed to clarify the specific molecular mechanisms leading to increased TIA biosynthesis with CrGLK silencing. This is the first identification and characterization of GLK in C. roseus and the first investigation of how chloroplast retrograde signaling might regulate TIA biosynthesis. [ABSTRACT FROM AUTHOR]

Background and Aim: Lincomycin is an antibiotic used in broiler farming and is commonly combined with other substances to achieve synergistic and complementary effects on the antibacterial spectrum and mechanism. We developed a specific high-performance liquid chromatography (HPLC) method to measure lincomycin levels in broiler tissues. This study aimed to determine the lincomycin level in tissues and compare it with the minimum inhibitory concentration (MIC) and maximum residue limit (MRL) of certain pathogenic bacteria. Materials and Methods: Three groups of broiler chickens were involved in the study (n = 20 in each group): A control group without lincomycin treatment and two groups (each further divided into two sub-groups) that received oral lincomycin at a dose of 1 g/10 kg of body weight daily for 7 and 14 consecutive days. Tissue samples were collected from each group 1 day and 1 week after lincomycin administration (ALA). This study validated the development of a technique for analyzing drug level degradation in tissues using HPLC. Descriptive and statistical analyses were performed for drug levels to assess their therapeutic value and safety based on lincomycin MIC of certain pathogenic bacteria and MRL. Results: The method validation resulted in linear regression and coefficient of determination for tissues with r² > 0.99, with a recovery rate of 90%-110%, precision as the coefficient of variation 15%, and specificity with no peak overlap for lincomycin. The limits of detection for the liver and kidney were 0.01 μg/g, 0.05 µg/g, and 0.1 µg/g for the breast muscle and all tissues. Administration of lincomycin for 7 and 14 days resulted in therapeutic value concentrations. Lincomycin levels in the liver and kidney of ALA exceeded the MRL, whereas breast muscles were below the MRL for a week of ALA treatment. Conclusion: Administration of lincomycin for 7 and 14 consecutive days resulted in therapeutic value; however, after a week, most tissues showed high drug concentrations that exceeded the MRL. It is necessary to carefully consider the prolonged therapeutic dose of lincomycin in broilers. Antibiotic therapy must be guided in such a way as to protect the product from harmful residues. [ABSTRACT FROM AUTHOR]

Antibiotics in the aquatic environment are of great concern as novel contaminants. In this study, we investigated the occurrence, distribution, potential sources, and risk assessment of antibiotics in an interconnected river–wetland–lake system. Thirty-three target antibiotics, including sulfonamides (SAs), macrolides (MLs), fluoroquinolones (FQs), tetracyclines (TCs), and chloramphenicol (CLs) belong to five common groups of antibiotics, were tested from water samples collected in the Panlong River, Xinghai Wetland, and Lake Dian (or Dianchi). Mass spectrophotometry was used to detect the target antibiotics, and the water quality parameters were measured in situ. We found four antibiotics, lincomycin (LIN), trimethoprim (TMP), sulfamethoxazole (SMX), and ofloxacin (OFL), with relatively low concentrations at the ng/L level, and detection rates among sample sites ranged from 42.3% to 76.9%, with maximum concentrations of 0.71 ng/L~5.53 ng/L. TMP was not detected in the Panlong River but appeared in the wetlands and Lake Dian. Midstream urban areas of the Panlong River showed the highest pollution among sites. Antibiotic concentrations were positively correlated with total nitrogen (TN) (p < 0.05) and showed some negative correlation with pH, salinity, and DO. According to the risk assessment, antibiotics in water do not pose a threat to human health and aquatic ecosystems, but a potentially harmful combined effect cannot be excluded. Our research offers a geographical summary of the distribution of antibiotics in urban river, wetland, and lake ecosystems in the plateau (PWL), which is important for predicting the distribution characteristics of antibiotics in the plateau water environment and establishing a standardized antibiotic monitoring and management system for the government. [ABSTRACT FROM AUTHOR]

The biosynthesis of bioactive secondary metabolites, specifically antibiotics, is of great scientific and economic importance. The control of antibiotic production typically involves different processes and molecular mechanism. Despite numerous efforts to improve antibiotic yields, joint engineering strategies for combining genetic manipulation with fermentation optimization remain finite. Lincomycin A (Lin-A), a lincosamide antibiotic, is industrially fermented by Streptomyces lincolnensis. Herein, the leucine-responsive regulatory protein (Lrp)-type regulator SLCG_4846 was confirmed to directly inhibit the lincomycin biosynthesis, whereas indirectly controlled the transcription of SLCG_2919, the first reported repressor in S. lincolnensis. Inactivation of SLCG_4846 in the high-yield S. lincolnensis LA219X (LA219XΔ4846) increases the Lin-A production and deletion of SLCG_2919 in LA219XΔ4846 exhibits superimposed yield increment. Given the effect of the double deletion on cellular primary metabolism of S. lincolnensis, Plackett-Burman design, steepest ascent and response surface methodologies were utilized and employed to optimize the seed medium of this double mutant in shake flask, and Lin-A yield using optimal seed medium was significantly increased over the control. Above strategies were performed in a 15-L fermenter. The maximal yield of Lin-A in LA219XΔ4846-2919 reached 6.56 g/L at 216 h, 55.1 % higher than that in LA219X at the parental cultivation (4.23 g/L). This study not only showcases the potential of this strategy to boost lincomycin production, but also could empower the development of high-performance actinomycetes for other antibiotics.

In this work, the supersensitive and selective determination of lincomycin (Lin) was achieved using a novel electroluminescent (ECL) aptasensor based on the synergistic integration of gold functionalized upconversion nanoparticles (UCNPs) and thiolated 3,4,9,10-perylene tetracarboxylic acid (PTCA). The integration of two luminophores of UCNPs and PTCA combined the merits of the cathodoluminescence stability of UCNPs and the high quantum yield of PTCA, which significantly promoted the ECL signal and analytical performance of the proposed sensor. The introduction of gold nanoparticles in UCNPs can not only improve the conductivity and ECL performance of UCNPs but also cause them to easily integrate with thiolated PTCA (t-PTCA) via an Au-S bond. The ECL signal of UCNPs@Au/t-PTCA/GCE was almost twice as strong as that of t-PTCA/GCE and tenfold higher than that of UCNPs@Au/GCE. Because of the non-conductive protein of the Lin aptamer, the ECL intensity of apt/UCNPs@Au/t-PTCA/GCE noticeably decreased. In the presence of Lin, the aptamer was pulled down from the sensing interface, resulting in the recovery of the ECL intensity of the sensor. Under optimal conditions, our proposed sensor can quantify the concentration of Lin in the range from 1.0 × 10−15 to 1.0 × 10−7 M with a low detection limit of 2.4 × 10−16 M (S/N = 3), exhibiting high sensitivity and specificity for the determination of Lin. [ABSTRACT FROM AUTHOR]

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Background: Antibiotics biosynthesis is usually regulated by the cluster-situated regulatory gene(s) (CSRG(s)), which directly regulate the genes within the corresponding biosynthetic gene cluster (BGC). Previously, we have demonstrated that LmbU functions as a cluster-situated regulator (CSR) of lincomycin. And it has been found that LmbU regulates twenty non-lmb genes through comparative transcriptomic analysis. However, the regulatory mode of CSRs' targets outside the BGC remains unknown. Results: We screened the targets of LmbU in the whole genome of Streptomyces lincolnensis and found fourteen candidate targets, among which, eight targets can bind to LmbU by electrophoretic mobility shift assays (EMSA). Reporter assays in vivo revealed that LmbU repressed the transcription of SLINC_0469 and SLINC_1037 while activating the transcription of SLINC_8097. In addition, disruptions of SLINC_0469, SLINC_1037, and SLINC_8097 promoted the production of lincomycin, and qRT-PCR showed that SLINC_0469, SLINC_1037, and SLINC_8097 inhibited transcription of the lmb genes, indicating that all the three regulators can negatively regulate lincomycin biosynthesis. Conclusions: LmbU can directly regulate genes outside the lmb cluster, and these genes can affect both lincomycin biosynthesis and the transcription of lmb genes. Our results first erected the cascade regulatory circuit of LmbU and regulators outside lmb cluster, which provides the theoretical basis for the functional research of LmbU family proteins. [ABSTRACT FROM AUTHOR]

Over 35 days feeding trial, the effects of lincomycin (LM) and butyrate glycerides (BG) supplementation on the growth performance, blood biochemical changes, immune response, and nutrient absorption related gene expression in broiler chickens were evaluated. A total of 480 one- day-old broiler chicks (Ross 308) were allotted into 6 groups (8 replicates/group with 10 birds/replicate). Experimental groups were arranged as follows; control group (A) fed the basal diet (BD); B, C, and D groups were fed the BD supplemented with 350, 500 and 1000 g BG/ton of feed respectively. While group E was fed the BD with 350 + 500 + 1000 g BG/ton of feed during starter, grower, and finisher periods, respectively, and finally group F which received lincomycin (LM) antibiotic (100 g/ton) in overall periods. Broilers that received BG in group E (birds supplemented with BG at 350 + 500 + 1000 g/ton feed) and LM (F) supplements both gained more body weight and weight growth (P<0.05). Lower feed intake and improved fcr were observed in all the experimental groups that received either BG or lm compared to control birds (P<0.05). Lower serum total cholesterol, triglycerides, and glutamic oxaloacetic transaminase (GOT) (P<0.05) were recorded in the lm or BG supplemented birds compared to the control. Higher immune response represented by enhanced antibody titer against H9N1, and newcastle disease (ND) and lower hepatic malondialdehyde (MDA) was obtained in groups D, E and F (P<0.05) compared to the other groups. Higher gene expression of GSH-Px, Glut 2, PepT1 and FAS was obtained in groups D, E and F (P<0.05) compared to the control. Supplemental BG or LM increased the unsaturated fatty acids (oleic, linoleic and linolenic acids) while lowered the palmitic acid contents in breast muscle (P<0.05) compared to control. In conclusion, supplementing broilers' diets with BG or lincomycin improved their growth performance, immune response and muscle fatty acid profile as well as modulatory role in lipid metabolism. Supplementing starter, grower, and finisher broiler diets with BG at 350 + 500 + 1000 g/ton each may be a useful substitute for lincomycin. [ABSTRACT FROM AUTHOR]

The transcription factor (TF)‐mediated regulatory network controlling lincomycin production in Streptomyces lincolnensis is yet to be fully elucidated despite several types of associated TFs having been reported. SLCG_2919, a tetracycline repressor (TetR)‐type regulator, was the first TF to be characterized outside the lincomycin biosynthetic cluster to directly suppress the lincomycin biosynthesis in S. lincolnensis. In this study, improved genomic systematic evolution of ligands by exponential enrichment (gSELEX), an in vitro technique, was adopted to capture additional SLCG_2919‐targeted sequences harboring the promoter regions of SLCG_6675, SLCG_4123‐4124, SLCG_6579, and SLCG_0139‐0140. The four DNA fragments were confirmed by electrophoretic mobility shift assays (EMSAs). Reverse‐transcription quantitative polymerase chain reaction (RT‐qPCR) showed that the corresponding target genes SLCG_6675 (anthranilate synthase), SLCG_0139 (LysR family transcriptional regulator), SLCG_0140 (beta‐lactamase), SLCG_6579 (cytochrome P450), SLCG_4123 (bifunctional DNA primase/polymerase), and SLCG_4124 (magnesium or magnesium‐dependent protein phosphatase) in ΔSLCGL_2919 were differentially increased by 3.3‐, 4.2‐, 3.2‐, 2.5‐, 4.6‐, and 2.2‐fold relative to those in the parental strain S. lincolnensis LCGL. Furthermore, the individual inactivation of these target genes in LCGL reduced the lincomycin yield to varying degrees. This investigation expands on the known DNA targets of SLCG_2919 to control lincomycin production and lays the foundation for improving industrial lincomycin yields via genetic engineering of this regulatory network. [ABSTRACT FROM AUTHOR]

Ribosome‐associated GTPases are conserved enzymes that participate in ribosome biogenesis and ribosome function. In bacteria, recent studies have identified HflX as a ribosome‐associated GTPase that is involved in both ribosome biogenesis and recycling under stress conditions. Plants possess a chloroplastic HflX homolog, but its function remains unknown. Here, we characterized the role of HflX in the plant Arabidopsis thaliana. Our findings show that HflX does not affect normal plant growth, nor does it play an essential role in acclimation to several different stresses, including heat, manganese, cold, and salt stress under the conditions tested. However, we found that HflX is required for plant resistance to chloroplast translational stress mediated by the antibiotic lincomycin. Our results suggest that HflX is a chloroplast ribosome‐associated protein that may play a role in the surveillance of translation. These findings provide new insight into the function of HflX as a ribosome‐associated GTPase in plants and highlight the importance of investigating conserved proteins in different organisms to gain a comprehensive understanding of their biological roles. [ABSTRACT FROM AUTHOR]

The Actinomycetes Streptomyces lincolnensis is the producer of lincosamide-type antibiotic lincomycin, a widely utilized drug against Gram-positive bacteria and protozoans. In this work, through gene knockout, complementation, and overexpression experiments, we identified LcbR1 (SLINC_1595), a GntR family transcriptional regulator, as a repressor for lincomycin biosynthesis. Deletion of lcbR1 boosted lincomycin production by 3.8-fold, without obvious change in morphological development or cellular growth. The homologues of LcbR1 are widely distributed in Streptomyces. Heterologous expression of SCO1410 from Streptomyces coelicolor resulted in the reduction of lincomycin yield, implying that the function of LcbR1 is conserved across different species. Alignment among sequences upstream of lcbR1 and their homologues revealed a conserved 16-bp palindrome (-TTGAACGATCCTTCAA-), which was further proven to be the recognition motif of LcbR1 by electrophoretic mobility shift assays (EMSAs). Via this motif, LcbR1 suppressed the transcription of lcbR1 and SLINC_1596 sharing the same bi-directional promoter. SLINC_1596, one important target of LcbR1, exerted a positive effect on lincomycin production. As detected by quantitative real-time PCR (qRT-PCR) analyses, the expressions of all selected structural (lmbA, lmbC, lmbJ, lmbV, and lmbW), resistance (lmrA and lmrB) and regulatory genes (lmrC and lmbU) from lincomycin biosynthesis cluster were upregulated in deletion strain ΔlcbR1 at 48 h of fermentation, while the mRNA amounts of bldD, glnR, ramR, SLCG_Lrp, and SLCG_2919, previously characterized as the regulators on lincomycin production, were decreased in strain ΔlcbR1, although the regulatory effects of LcbR1 on the above differential expression genes seemed to be indirect. Besides, indicated by EMSAs, the expression of lcbR1 might be regulated by GlnR, SLCG_Lrp, and SLCG_2919, which shows the complexity of the regulatory network on lincomycin biosynthesis. Key points: • LcbR1 is a novel and conservative GntR family regulator regulating lincomycin production. • LcbR1 modulates the expressions of lcbR1 and SLINC_1596 through a palindromic motif. • GlnR, SLCG_Lrp, and SLCG_2919 can control the expression of lcbR1. [ABSTRACT FROM AUTHOR]

Antimicrobial resistance is considered a problem for public health globally. New forms of resistance mechanisms are developing and spreading daily around the world. For that reason, a potential method for overcoming the antimicrobial resistance of several pathogens that cause deadly infections is the use of silver nanoparticles. In the present study, silver nanoparticles (AgNPs) and AgNPs conjugated with lincomycin were synthesized and characterized. The antimicrobial activity of AgNPs alone and after conjugating with lincomycin was also evaluated. Transmission electron microscopy (TEM) and scanning electron microscopy (SEM) were employed to determine the morphological properties of AgNPs and AgNPs-lincomycin, which showed the average mean size was ±26.73 nm for AgNPs and ±28.31 nm for AgNPslincomycin with a spherical shape. In addition, the dynamic light scattering (DLS) and zeta potential were assessed. The results showed an improvement in the antimicrobial activity of lincomycin after conjugation with AgNPs, as the inhibition zone diameter reached 45.66±1.52 mm for Staphylococcus warneri, 17±2 mm for Serratia marcescens, and 22.33±1.52 mm for Candida guilliermondii. It also showed a synergistic effect of AgNPs-lincomycin against the biofilm formation of Candida guilliermondii, whereas Serratia marcescens was slightly affected. The MIC of AgNPs was 25 μg/ml for Staphylococcus warneri and Candida guilliermondii, whereas 100 μg/ml for Serratia marcescens, when the MIC of AgNPs-lincomycin was 12.5 μg/ml for Staphylococcus warneri and Candida guilliermondii, and 100 μg/ml for Serratia marcescens. In the experiment of the effect of AgNPs on gene expression of the antibiotic resistance genes, including the blaZ gene in Staphylococcus warneri, the aac(6´)-Ib-cr gene in Serratia marcescens, and the CDR1 gene in Candida guilliermondii, the results showed that there was a decrease in gene expression after being treated with AgNPs in each gene. [ABSTRACT FROM AUTHOR]

Lincomycin is a widely used aminoglycoside antibiotic. For its separation from fermentation broth in production, solvent extraction is usually applied because of its low cost and high capacity compared to other bioseparation methods. The multistage mixer-settler is a common extraction equipment in commercial production, but it occupies a large area and causes pollution. In this study, a fully enclosed turbine tower was designed and applied in order to replace the mixer-settler. Its structure parameters (turbine diameter, tray porosity) were optimized on the basis of the extraction effect of lincomycin. The results showed that with 35% tray porosity and 28/26 mm turbine diameter of the tower, the extraction rate was kept above 99.0% steadily under 375 rpm/min rotating speed and 60 °C temperature. The extraction effect is much better than mixer-settler and such turbine tower is expected to be applied in the commercial production of lincomycin. [ABSTRACT FROM AUTHOR]

The antibiotic lincomycin binds to the 23S ribosomal RNA peptidyl transferase loop region to inhibit protein synthesis. However, lincomycin can also stimulate the growth and secondary metabolism of actinomycetes in a concentrationdependent manner. In Streptomyces coelicolor A3(2), lincomycin stimulates the production of the blue-pigmented antibiotic actinorhodin at concentrations below the minimum inhibitory concentration. To better understand the molecular mechanism underlying these concentration-dependent positive effects, this study investigated how the target molecule, the ribosome, undergoes dynamic changes in the presence of lincomycin and explored the ribosome-related factors involved. Lincomycin, at a concentration that stimulates actinorhodin production of S. coelicolor A3(2), could restore temporarily arrested ribosome function by utilizing ribosome-related proteins and translation factors, presumably under the control of the transcription factor WblC protein that confers intrinsic resistance to multiple translation-inhibiting antibiotics, to eventually produce stable and active ribosomes even during the late growth phase. This qualitatively and quantitatively positive ribosome alteration can be advantageous for producing actinorhodin biosynthetic enzymes. A series of gene expression and biochemical analyses revealed that lincomycin at the concentration that induces ribosomal stabilization in S. coelicolor A3(2) could influence the localization of the 20S proteasome-related proteins, resulting in reduced proteasome activity. These findings suggest that the functional analysis of 20S proteasome represents a potential pivotal challenge for understanding the molecular mechanism of ribosome stabilization induced by lincomycin. Therefore, as lincomycin can dynamically alter its target molecule, the ribosome, we discuss the future issues and prospects for an increased understanding of the concentration-dependent properties of antibiotics. [ABSTRACT FROM AUTHOR]

Lincomycin, a clinically important antibiotic against Gram-positive bacteria, is a sulfur-containing metabolite produced by Streptomyces lincolnensis. The main problem in the fermentation is the high content of lincomycin B, which is a by-product of insufficient methylation by S-adenosylmethionine (SAM) dependent methyltransferase. In this study, the content of lincomycin B was decreased from 4.04% to 0.75% and from 8.20% to 5.78% by the addition of NaCl as an osmotic regulator in flask and 15 L bioreactor fermentation, respectively. By analysis of the sulfur-containing metabolites (cysteine, homocysteine, SAM, and ergothioneine) and high-throughput RNA sequencing, the mechanism of osmotic regulation on lincomycin fermentation was studied. The results showed that the synthesis of methyl donor (SAM) and sulfur donor of lincomycin (ergothioneine and mycothiol) were increased at the metabolite level and transcription level under osmotic stimulation; the transcription of genes involved in sulfur assimilation (tauA, ssuA1B1C1) and lincomycin biosynthesis were also up-regulated significantly. This study provides a cost-effective method to reduce lincomycin B and increase the production of lincomycin A. The mechanisms of osmotic pressure regulation on lincomycin fermentation was also elucidated, which may provide more genetically modified targets for overproduction of lincomycin and other sulfur containing secondary metabolites. [ABSTRACT FROM AUTHOR]

ABSTRACT: Lincomycin (LCM) is an antibiotic used to treat severe bacterial infections in livestock and companion animals. In this study, we aimed to investigate the oral bioavailability of LCM with PK data after IV and PO administration and to compare differences in drug residue patterns in eggs. To ensure food safety, an additional study on egg residue was conducted using 3 different commercial LCM drugs. For bioavailability study, laying hens were divided into oral and intravenous (n = 8/group) groups and received single dose (10 mg/kg) of LCM. The limits of quantification for LCM were 0.729 μg/mL and 0.009 mg/kg in plasma and eggs, respectively. The oral group exhibited a significantly lower average serum drug concentration than the IV group, with a bioavailability of 2.6%. Furthermore, the egg residue profiles confirmed reduced systemic drug exposure after oral administration. For the commercial LCM drug egg residue experiment, laying hens were divided into low- and high-dose groups (n = 12/group) for each drug and treated with the recommended dosage and administration method for each respective drug. The eggs were collected and analyzed until 14 d after the last drug treatment. Despite differences in the LCM content and formulation among commercial drugs, all the tested commercial drugs showed average concentrations below the MRL in eggs within approximately 3 d after the last drug treatment. In this study, we have confirmed that LCM has a low oral absorption rate in laying hens, and this was consistent with the findings from the egg residue profiles. Further studies are requested to elucidate the exact reasons for evidently low oral drug absorption in laying hens.

The excessive accumulation of antibiotic residue in animal-derived foods pose a threat to our heath. Therefore, it is necessary to establish a rapid, simple, effective, safe and highly sensitive lincomycin (LIN) residues detection method by high-performance liquid chromatography equipped with ultraviolet. Here, due to the weak UV absorption of LIN, the p-Toluenesulfonyl isocyanate (PTSI, has a strong UV absorption) was binded to the hydroxyl group of LINs by using pre-column derivatization method, improving the UV absorption of LIN. This 9-min-run analytical method detects and quantifies LIN residues with acceptable validation performance parameters in terms of sensitivity, selectivity, linearity, the limit of quantification, accuracy, and precision.Our result showed that the limits of detection and quantification for LIN in samples were 25–40 μg/kg and 40–60 μg/kg, respectively, under the optimized conditions. Furthermore, the LIN recoveries ranged from 72.4 % to 96.7 %; the determination coefficients were 0.9912–0.9996, and the relative standard deviations were 5.6 %-12.7 %. Therefore, the HPLC-UVD method for detecting LIN residues in animal-derived food products has been established.

The heterogeneous Fenton process is a strategy for overcoming the greatest shortcomings of traditional homogeneous Fenton, i.e. the high generation of ferric hydroxide sludge and effectivity in a limited pH range. In this study, we constructed a heterogeneous Fenton system with natural iron-bearing clay mineral (nontronite) and dimethoxyhydroquinone (DMHQ) to degrade lincomycin (LCM) without the addition of H2O2. The degradation mechanism was derived from the hydroxyl radicals (•OH) produced from the oxygenation of Fe(II) in nontronites, which was reduced by DMHQ. Acidic conditions and low concentrations of LCM were favourable for LCM degradation. When the solution pH increased from 3 to 7, the final LCM removal ratio decreased from 95 to 46%. However, LCM can still be degraded by 46% under neutral conditions and 20% at the LCM concentration of 500 μmol/L. The nontronite has good reusability, and the LCM degradation efficiency in the fourth cycle still exceeded 90% of the original efficiency. The degradation sites of LCM mainly occurred in the methyl thioether moiety and the aliphatic amine group on the pyrrolidine ring, with the final product of CO2. This research presents a new eco-friendly and cost-effective method for the heterogenous Fenton process without external H2O2. HIGHLIGHTS 2,6-DMHQ could reduce structural Fe(III) to form Fe(II) to react with O2 and yield •OH.; LCM could be effectively degraded in the nontronite/2,6-DMHQ reaction system.; The nontronite was efficient with good reusability material.; The clay mineral not only provides iron but also a constrained reaction environment.; The reaction system is efficient, simple, and does not need H2O2 addition.;

BackgroundLincomycin, a natural antibiotic, is widely used by animal and fishery husbandries to prevent infections and treat diseases. It endangers people’s health when they eat foods containing lincomycin residue, especially the frequent consumption of milk and chicken products containing lincomycin. Hence, it is extremely important to evaluate the content of lincomycin in food samples. However, a direct analysis of lincomycin in milk and chicken is quite difficult because of its very low concentration level and the presence of undesirable matrix effects. Therefore, selective and efficient extraction of lincomycin from complex food samples prior to its quantification is required.ResultsIn this study, lincomycin-imprinted silica nanoparticles were prepared according to boronate affinity-based template-immobilized surface imprinting. Silica nanoparticles and boronic acid ligands 3-fluoro-4-formylphenylboronic acid were used as supporting materials and functional monomers, respectively. The prepared lincomycin-imprinted silica nanoparticles exhibited several significant results, such as good specificity, high binding capacity (19.45 mg/g), fast kinetics (6 min), and low binding pH (pH 5.0) toward lincomycin. The reproducibility of lincomycin-imprinted silica nanoparticles was satisfactory. The lincomycin-imprinted silica nanoparticles could still be reused after seven adsorption–desorption cycles, which indicated high chemical stability. In addition, the recoveries of the proposed method for lincomycin at three spiked levels of analysis in milk and chicken were 93.3–103.3% and 90.0–100.0%, respectively.ConclusionThe prepared lincomycin-imprinted silica nanoparticles are feasible for the recognition of target lincomycin with low concentrations in real food samples such as milk and chicken. Our approach makes sample pre-preparation simple, fast, selective, and efficient.

Background: Sodium butyrate (SB) is a short-chain fatty acid and a safe antibiotic alternative. During 35 days, this study compared the impact of coated SB (Butirex C4) and lincomycin (Lincomix) on broiler growth, gut health, and litter hygiene in 1200 one-day-old Ross-308 broiler chicks that were randomly assigned into 5-dietary groups with 5-replications each. Groups divided as follows: T1: Basal diet (control), T2: Basal diet with buffered SB (1 kg/ton starter feed, 0.5 kg/ton grower-finisher feeds), T3: Basal diet with 100 g/ton lincomycin, T4: Basal diet with buffered SB (0.5 kg/ton starter feed, 0.25 kg/ton grower-finisher feeds) + 50 g/ton lincomycin, and T5: Basal diet with buffered SB (1 kg/ton starter feed, 0.5 kg/ton grower-finisher feeds) + 50 g/ton lincomycin. Birds were housed in a semi-closed deep litter house, where feed and water were available ad libitum. Results were statistically analyzed using ANOVA and Tukey's post hoc tests. Results: Combined dietary supplementation with SB and lincomycin (T4 and T5) significantly enhanced body weights, weight gains, feed conversion ratio, and profitability index. Also, carcasses in T4 and T5 exhibited the highest dressing, breast, thigh, and liver yields. T5 revealed the best blood biochemical indices, while T3 showed significantly elevated liver and kidney function indices. T4 and T5 exhibited the highest expression levels of IGF-1 and TLR4 genes, the greatest villi length of the intestinal mucosa, and the lowest levels of litter moisture and nitrogen. Clostridia perfringens type A alpha-toxin gene was confirmed in birds' caeca, with the lowest clostridial counts defined in T4. Conclusions: Replacing half the dose of lincomycin (50 g/ton) with 0.5 or 1 kg/ton coated SB as a dietary supplement mixture showed the most efficient privileges concerning birds' performance and health. [ABSTRACT FROM AUTHOR]

Lincomycin (LC) is an extensively applied broad-spectrum antibiotic, and its considerable residues in wastewater have caused a series of environmental problems, which makes degradation of LC wastewater extremely urgent. In this work, we have constructed a novel boron nitride (BN) and samarium (Sm) co-modified Ti/PbO2 as anode for high-performance degradation of LC wastewater. Compared with Ti/PbO2, Ti/PbO2-Sm, and Ti/PbO2-BN electrodes, Ti/PbO2-BN-Sm electrode with smaller pyramidal particles possesses higher oxygen evolution potential (2.32 V), excellent accelerated service life (103 h), and outstanding electrocatalytic activity. The single-factor experiments demonstrate that under optimized conditions (current density of 20 mA.cm-2, 6.0 g L-1 Na2SO4, pH 9, and temperature of 30°C), removal rate and COD degradation rate of LC at 3 h have reached 92.85% and 89.11%, respectively. At the same time, degradation of LC is in accordance with the primary kinetic model. Based on the analysis of high-performance liquid chromatography-mass spectrometry (HPLC-MS), four possible degradation pathways are hypothesized. Therefore, efficient electrochemical degradation of LC by using an extremely long-life Ti/PbO2 electrode with high catalytic activity may be a promising method. [ABSTRACT FROM AUTHOR]

Antibiotic drug residues can adversely affect the human body. Lincomycin is a common veterinary drug that can form residues in foods of animal origin. However, the detection of trace residue levels of lincomycin residues in real samples is challenging. Here, a simple solid phase extraction (SPE) method was developed for the enrichment of lincomycin from cow milk samples before its detection by high-performance liquid chromatography-tandem mass spectrometry (HPLC-MS/MS). The adsorbent used in the SPE was a Cu-based metal-organic framework (Cu-MOF) prepared by the solvothermal synthesis approach. The prepared MOFs were characterized using scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), X-ray diffractometry (XRD), differential thermogravimetric analysis (TG-DTA), and N2 adsorption-desorption experiments. The adsorption capacity (adsorption equilibrium, extraction time, pH), and elution solvent parameters were investigated. Under the optimized conditions of the HPLC-MS/MS method, lincomycin was detected in the linear range of 10–200 g/L with a detection limit of 0.013 ng/mL. Commercial milk samples were spiked with lincomycin, and a recovery rate between 92.3% and 97.2% was achieved. Therefore, the current method can be successfully applied for the enrichment and determination of lincomycin from milk samples. [ABSTRACT FROM AUTHOR]

AtrA belongs to the TetR family and has been well characterized for its roles in antibiotic biosynthesis regulation. Here, we identified an AtrA homolog (AtrA-lin) in Streptomyces lincolnensis. Disruption of atrA-lin resulted in reduced lincomycin production, whereas the complement restored the lincomycin production level to that of the wild-type. In addition, atrA-lin disruption did not affect cell growth and morphological differentiation. Furthermore, atrA-lin disruption hindered the transcription of regulatory gene lmbU, structural genes lmbA and lmbW inside the lincomycin biosynthesis gene cluster, and 2 other regulatory genes, adpA and bldA. Completement of atrA-lin restored the transcription of these genes to varying degrees. Notably, we found that AtrA-lin directly binds to the promoter region of lmbU. Collectively, AtrA-lin positively modulated lincomycin production via both pathway-specific and global regulators. This study offers further insights into the functional diversity of AtrA homologs and the mechanism of lincomycin biosynthesis regulation. [ABSTRACT FROM AUTHOR]

Background and Aims The photoprotective role of foliar anthocyanins has long been ambiguous: exacerbating, being indifferent to or ameliorating the photoinhibition of photosynthesis. The photoinhibitory light spectrum and failure to separate photo-resistance from repair, as well as the different methods used to quantify the photo-susceptibility of the photosystems, could lead to such a discrepancy. Methods We selected two congeneric deciduous shrubs, Prunus cerasifera with anthocyanic leaves and Prunus triloba with green leaves, grown under identical growth conditions in an open field. The photo-susceptibilities of photosystem II (PSII) and photosystem I (PSI) to red light and blue light, in the presence of lincomycin (to block the repair), of exposed leaves were quantified by a non-intrusive P700+ signal from PSI. Leaf absorption, pigments, gas exchange and Chl a fluorescence were also measured. Key Results The content of anthocyanins in red leaves (P. cerasifera) was >13 times greater than that in green leaves (P. triloba). With no difference in maximum quantum efficiency of PSII photochemistry (F v/ F m) and apparent CO2 quantum yield (AQY) in red light, anthocyanic leaves (P. cerasifera) showed some shade-acclimated suites, including lower Chl a / b ratio, lower photosynthesis rate, lower stomatal conductance and lower PSII/PSI ratio (on an arbitrary scale), compared with green leaves (P. triloba). In the absence of repair of PSII, anthocyanic leaves (P. cerasifera) showed a rate coefficient of PSII photoinactivation (k i) that was 1.8 times higher than that of green leaves (P. triloba) under red light, but significantly lower (−18 %) under blue light. PSI of both types of leaves was not photoinactivated under blue or red light. Conclusions In the absence of repair, anthocyanic leaves exhibited an exacerbation of PSII photoinactivation under red light and a mitigation under blue light, which can partially reconcile the existing controversy in terms of the photoprotection by anthocyanins. Overall, the results demonstrate that appropriate methodology applied to test the photoprotection hypothesis of anthocyanins is critical. [ABSTRACT FROM AUTHOR]

In this study, we evaluated the biotransformation mechanisms of lincomycin (LIN) and three fluoroquinolone antibiotics (FQs), ciprofloxacin (CFX), norfloxacin (NFX), and ofloxacin (OFX), which regularly enter aquatic environments through human activities, by different ammonia-oxidizing microorganisms (AOM). The organisms included a pure culture of the complete ammonia oxidizer (comammox) Nitrospira inopinata, an ammonia oxidizing archaeon (AOA) Nitrososphaera gargensis, and an ammonia-oxidizing bacterium (AOB) Nitrosomonas nitrosa Nm90. The removal of these antibiotics by the pure microbial cultures and the protein-normalized biotransformation rate constants indicated that LIN was significantly co-metabolically biotransformed by AOA and comammox, but not by AOB. CFX and NFX were significantly co-metabolized by AOA and AOB, but not by comammox. None of the tested cultures transformed OFX effectively. Generally, AOA showed the best biotransformation capability for LIN and FQs, followed by comammox and AOB. The transformation products and their related biotransformation mechanisms were also elucidated. i) The AOA performed hydroxylation, S-oxidation, and demethylation of LIN, as well as nitrosation and cleavage of the piperazine moiety of CFX and NFX; ii) the AOB utilized nitrosation to biotransform CFX and NFX; and iii) the comammox carried out hydroxylation, demethylation, and demethylthioation of LIN. Hydroxylamine, an intermediate of ammonia oxidation, chemically reacted with LIN and the selected FQs, with removals exceeding 90%. Collectively, these findings provide important fundamental insights into the roles of different ammonia oxidizers and their intermediates on LIN and FQ biotransformation in nitrifying environments including wastewater treatment systems.

In order to preliminarily explore the distribution of antibiotic pollution in the coastal waters of eastern China, the concentrations of 13 antibiotics in 5 representative coastal rivers in Jiangsu and 21 sampling sites in the coastal waters of Jiangsu were analyzed. The total antibiotic concentrations in the 5 rivers ranged from 33.14 to 417.78 ng L−1, and the total antibiotic concentrations in the 21 sampling sites ranged from 0.90 to 86.33 ng L−1. Macrolides exhibited the highest total concentration and the maximum detection frequency in both coastal rivers and the coastal waters. The concentrations of antibiotics in a sampling site decreased as the distance of the sampling site from the coastline increased, indicating that river inputs are important sources of antibiotic pollution in the coastal waters of Jiangsu. The detection frequencies of roxithromycin, lincomycin, azithromycin, and sulfamethoxazole in the rivers and sampling sites were above 70%. Correlation analysis showed that the concentrations of antibiotics were positively correlated with the levels of chemical oxygen demand, total phosphorus, and total nitrogen. Risk assessments revealed that roxithromycin and ofloxacin posed medium ecological and resistance risks, respectively, to the most sensitive aquatic organisms in the coastal waters of Jiangsu. The results of this study highlight the significance of monitoring and controlling the concentrations of antibiotic contaminants in the coastal waters of Jiangsu. [ABSTRACT FROM AUTHOR]

Solvent exchange-induced in situ forming gel (ISG) has emerged as a versatile drug delivery system, particularly for periodontal pocket applications. In this study, we developed lincomycin HCl-loaded ISGs using a 40% borneol-based matrix and N-methyl pyrrolidone (NMP) as a solvent. The physicochemical properties and antimicrobial activities of the ISGs were evaluated. The prepared ISGs exhibited low viscosity and reduced surface tension, allowing for easy injection and spreadability. Gel formation increased the contact angle on agarose gel, while higher lincomycin HCl content decreased water tolerance and facilitated phase separation. The drug-loading influenced solvent exchange and matrix formation, resulting in thinner and inhomogeneous borneol matrices with slower gel formation and lower gel hardness. The lincomycin HCl-loaded borneol-based ISGs demonstrated sustained drug release above the minimum inhibitory concentration (MIC) for 8 days, following Fickian diffusion and fitting well with Higuchi's equation. These formulations exhibited dose-dependent inhibition of Staphylococcus aureus ATCC 25923, Escherichia coli ATCC 8739, and Prophyromonas gingivalis ATCC 33277, and the release of NMP effectively inhibited Candida albicans ATCC 10231. Overall, the 7.5% lincomycin HCl-loaded 40% borneol-based ISGs hold promise as localized drug delivery systems for periodontitis treatment. [ABSTRACT FROM AUTHOR]

Lincomycin is one of the most important antibiotics. However, transcriptional regulation network of secondary metabolism in Streptomyces lincolnensis, the lincomycin producer, remained obscure. AdpA from S. lincolnensis (namely AdpAlin) has been proved to activate lincomycin biosynthesis. Here we found that both lincomycin and melanin took l‐tyrosine as precursor, and AdpAlin activated melanin biosynthesis as well. Three tyrosinases, MelC2, MelD2, and MelE, and one tyrosine peroxygenase, LmbB2, participated in lincomycin and melanin biosynthesis in different ways. For melanin biosynthesis, MelC2 was the only key enzyme required. For lincomycin biosynthesis, MelD2 and LmbB2 were positive factors and were suggested to convert l‐tyrosine to l‐dihydroxyphenylalanine (l‐DOPA). Otherwise, MelC2 and MelE were negative factors for lincomycin biosynthesis and they were supposed to oxidize l‐DOPA to generate melanin and certain unknown metabolite, respectively. Based on in silico analysis combined with electrophoretic mobility shift assays (EMSAs), we proved that AdpAlin directly interacted with promoters of melC, melD, and melE by binding to putative AdpA‐binding sites in vitro. Moreover, in vivo experiments revealed that AdpAlin positively regulated the transcription of melC and melE, but negatively regulated melD. In conclusion, AdpAlin was the switch of secondary metabolism in S. lincolnensis, and it modulated precursor flux of lincomycin and melanin biosynthesis by directly activating melC, melE, and lmbB1/lmbB2 or repressing melD. [ABSTRACT FROM AUTHOR]

The aim of the current study was to investigate the prevalence of Clostridium perfringens (C. perfringens) recovered from animal faeces, as well as to determine the antimicrobial susceptibility of such isolates. A total of 14 (14/100; 14%) C. perfringens isolates were isolated from the 100 analysed samples (twelve recovered from faecal samples collected from pigs and two from veal calves' faecal samples). The preponderant genotype was type A, with all isolates being cpa-positive. The most potent antimicrobial agents against C. perfringens proved to be vancomycin, rifampicin and lincomycin. A strong resistance to tetracycline (71.4%), penicillin (64.2%), erythromycin (42.8%) and enrofloxacin (35.7%) was also observed. To the best of our knowledge, this is the first analysis regarding the prevalence, characterization and antimicrobial susceptibility of C. perfringens in food-producing animals in Romania, adding further evidence for the probable role of animals as a source of resistant C. perfringens strains. [ABSTRACT FROM AUTHOR]

Lincomycin is a broad-spectrum antibiotic and particularly effective against Gram-positive pathogens. Albeit familiar with the biosynthetic mechanism of lincomycin, we know less about its regulation, limiting the rational design for strain improvement. We therefore analyzed two-component systems (TCSs) in Streptomyces lincolnensis, and selected eight TCS gene(s) to construct their deletion mutants utilizing CRISPR/Cas9 system. Among them, lincomycin yield increased in two strains (Δ3900–3901 and Δ5290–5291) while decreased in other four strains (Δ3415–3416, Δ4153–4154, Δ4985, and Δ7949). Considering the conspicuous effect, SLINC_5291-5290 (AflQ1-Q2) was subsequently studied in detail. Its repression on lincomycin biosynthesis was further proved by gene complementation and overexpression. By binding to a 16-bp palindromic motif, the response regulator AflQ1 inhibits the transcription of its encoding gene and the expression of eight operons inside the lincomycin synthetic cluster (headed by lmbA, lmbJ, lmbK, lmbV, lmbW, lmbU, lmrA, and lmrC), as demonstrated by quantitative RT-PCR and electrophoretic mobility shift assays. Besides, the regulatory genes including bldD, glnR, lcbR1, and ramR are also regulated by the TCS. According to the screening towards nitrogen sources, aspartate affects the regulatory behavior of histidine kinase AflQ2. And in return, AflQ1 accelerates aspartate metabolism via ask-asd, asd2, and thrA. In summary, we acquired six novel regulators related to lincomycin biosynthesis, and elucidated the regulatory mechanism of AflQ1-Q2. This highly conserved TCS is a promising target for the construction of antibiotic high-yield strains. Key points: • AflQ1-Q2 is a repressor for lincomycin production. • AflQ1 modulates the expression of lincomycin biosynthetic and regulatory genes. • Aspartate affects the behavior of AflQ2, and its metabolism is promoted by AflQ1. [ABSTRACT FROM AUTHOR]

In this study, an immunochromatographic test (using the Charm QUAD2® Test) was used to screen for residual macrolides and lincosamides in raw cow's milk. The validation parameters (selectivity/specificity, detection capability (CCβ), and ruggedness) were in agreement with the requirements of[EC] 2021. The selectivity of the immunochromatographic test was verified by the negative results of microbiological tests. The false-positive rate was 0%. The CCβ values of the immunochromatographic test for various antibiotics in milk were as follows: erythromycin 0.02 mg/kg, spiramycin 0.1 mg/kg, tilmicosin 0.025 mg/kg, tylosin 0.05 mg/kg, lincomycin 0.15 mg/kg, and pirlimycin 0.15 mg/kg. The determined CCβ values were lower than the respective maximum residue limits (MRLs; regulatory limits in Japan) for milk, except for lincomycin (equal to the MRL). The presence of antibiotic groups other than macrolides and lincosamides did not interfere with the specificity of the test. It showed no significant difference in lot-to-lot repeatability. The results obtained by the two researchers showed no significant differences. Finally, the test was applied to milk samples obtained from a tylosin-treated cow. The outcome was positive and in agreement with the results of the chemical analytical and microbiological methods. Therefore, this validated immunochromatographic test is expected to be suitable for routine analysis to ensure milk safety. [ABSTRACT FROM AUTHOR]

Lincomycin industrially produced by Streptomyces lincolnensis can be adopted to treat infections caused by Gram-positive bacteria. SLCG_Lrp, a transcriptional regulator of the Lrp family, was first identified to positively regulate lincomycin biosynthesis. However, the regulatory role of SLCG_Lrp is yet to be elucidated. This study utilized RNA-seq for comparing the transcriptome profile of original-strain LCGL and the ΔSLCGL_Lrp mutant. A total of 244 genes comprising 116 downregulated and 128 upregulated genes were differentially expressed between LCGL and ΔSLCGL_Lrp. An in-depth analysis revealed that SLCG_Lrp promotes nitrate assimilation but inhibits fatty acid metabolism, as well as directly regulates five regulators participating in the modulation of multiple cellular processes. With individual inactivation of those regulatory genes in S. lincolnensis LCGL, we confirmed the FadR transcriptional regulator SLCG_2185 was obviously correlated with lincomycin production and found it to transcriptionally stimulate the lincomycin biosynthetic cluster. Furthermore, SLCG_2185 overexpression in the high-yield S. lincolnensis LA219X promoted lincomycin production by 17.8%, and SLCG_2185 being co-overexpressed with SLCG_Lrp in LA219X increased lincomycin production by 28.1% compared to LA219X. Therefore, this investigation not only provides a direction for further investigations regarding the regulation mechanism of SLCG_Lrp, but also provides a basis for guiding the further improvement of lincomycin levels. [ABSTRACT FROM AUTHOR]

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Pharmaceutical plants are an essential source of antibiotics emitted into the aqueous environment. The monitoring of target antibiotics in pharmaceutical plants through various regions is vital to optimize contaminant release. The occurrence, distribution, removal, and ecological risk of 30 kinds of selected antibiotics in 15 pharmaceutical plants in the Pearl River Delta (PRD) were investigated in this study. Lincomycin (LIN) showed the highest concentration (up to 56,258.3 ng/L) in the pharmaceutical plant influents from Zhongshan city. Norfloxacin (NFX) showed a higher detection frequency than other antibiotics. In addition, the spatial distribution of antibiotics in pharmaceutical plants showed significant differences, with higher concentrations of total antibiotics found in pharmaceutical plant influents in Shenzhen City than those of different regions in PRD. The treatment processes adopted by pharmaceutical plants were commonly ineffective in removing antibiotics, with only 26.7% of antibiotics being effectively removed (average removal greater than 70%), while 55.6% of antibiotics had removal rates of below 60%. The anaerobic/anoxic/oxic (AAO)-membrane bioreactor (MBR) combined process exhibited better treatment performance than the single treatment process. Sulfamethoxazole (SMX), ofloxacin (OFL), erythromycin-H2O (ETM-H2O), sulfadiazine (SDZ), sulfamethazine (SMZ), norfloxacin (NFX), and ciprofloxacin (CIP) in pharmaceutical plant effluents posed high or moderate ecological risk and deserve particular attention. [ABSTRACT FROM AUTHOR]

Five hundred and sixty broilers were placed into seven groups. The first negative control (A) was fed the baseline diet (BD); the second positive control (B) was fed BD and exposed to clostridium infection; and the third and fourth groups (C, D) were the same as the positive control with 500 and 1000 g/ton of feed, respectively. The fifth group (E) served as a positive control for the addition of lincomycin, whereas the sixth and seventh groups (F, G) received the same designed diet as the E group but supplemented with HA at 500 and 1000 g/ton of feed, respectively. Birds infected with Clostridium perfringens had considerably decreased body weight. However, a dietary combination of HA and lincomycin resulted in a greater improvement in growth. Body weight increased after 35 days, but feed intake dropped, therefore HA and lincomycin supplementation enhanced feed conversion ratio. Supplementing with HA and lincomycin increased crude protein retention. Furthermore, these additions mitigated the detrimental effects of clostridial infection on the gut by reducing degenerative changes in intestinal villi and increasing villi length, particularly at higher HA/lincomycin doses. In conclusion, nutritional supplementation with humic acid and lincomycin improved blood biochemistry, and gut morphology in broilers infected with Clostridium difficile.

Neat stallion semen can contain a variety of microorganisms, some of which may impair sperm quality and/or cause infection of the mares' reproductive tract. For this reason, antibiotics are commonly added to semen extenders. A combination of gentamicin, tylosin, lincomycin and spectinomycin (GTLS) has been recommended for use, but there are no reports on the use of this mixture in equine semen extender. Penicillin and amikacin (PA) are safe for preserving sperm quality while effectively controlling bacterial growth in equine cooled stored semen, but data on frozen semen are scarce. Therefore, a bioequivalence study was performed to assess the bactericidal activity of GTLS and PA in equine frozen semen. Nine mature, healthy stallions were used in the study. Split ejaculates were processed using media without antibiotics (Control) or with different antibiotics. For the GTLS group, centrifugation medium and freezing extender were prepared with gentamicin 250 μg/ml, tylosin 50 μg/ml, lincomycin 150 μg/ml and spectinomycin 300 μg/ml. For the PA group, the centrifugation medium was prepared with potassium penicillin G (PPG) 1200 units/ml and the freezing extender was prepared with PPG 1200 units/ml and amikacin 500 μg/ml. Semen processed in extenders without antibiotics had higher (p <.005) bacterial loads throughout all cryopreservation processing steps than semen samples processed using antibiotics. There were no differences in semen bacterial load after centrifugation, 15 and 30 min after final extension, and after thawing between GTLS and PA groups, but PA had faster (p <.05) kill‐time kinetics than GTLS. Only minor differences in sperm kinetic parameters were observed among groups. In conclusion, this study demonstrated bioequivalence between GTLS and PA in mitigating end‐point bacterial loads. Prudent concentrations of the antibiotic mixtures evaluated in this study can be considered both effective and sperm‐safe for equine frozen semen. [ABSTRACT FROM AUTHOR]

Lincomycin produced by Streptomyces lincolnensis is a critical antibacterial antibiotic in the clinical. To further understand the regulatory mechanism of lincomycin biosynthesis, we identified an alternative σ factor, σLsl, in Streptomyces lincolnensis NRRL 2936. Deletion of sigLsl resulted in an increase in cell growth but a decrease in lincomycin production. σLsl boosted lincomycin biosynthesis by directly stimulating the transcription of four genes (lmbD, lmbV, lmrC, and lmbU) within the lincomycin biosynthetic lmb gene cluster. Besides, σLsl participated in lincomycin biosynthesis by directly stimulating the transcription of mshC, a gene responsible for MSH synthesis. In conclusion, our findings demonstrated that σLsl plays a direct regulatory role in lincomycin biosynthesis. This study extends the understanding of molecular mechanisms of lincomycin biosynthetic regulation. [ABSTRACT FROM AUTHOR]

Streptomyces albus J1074 is one of the most popular heterologous expression platforms among streptomycetes. Identification of new genes and mutations that influence specialized metabolism in this species is therefore of great applied interest. Here, we describe S. albus KO-1304 that was isolated as a spontaneous lincomycin-resistant variant of double rpsLR94GrsmGR15SG40E mutant KO-1295. Besides altered antibiotic resistance profile, KO-1304 exhibited increased antibiotic activity as compared to its parental strains. KO-1304 genome sequencing revealed mutations within gene XNR_2147 encoding putative TetR-like protein. Gene XNR_2146 for efflux protein is the most likely target of repressing action of Xnr_2147. Our data agree with the scenario where lincomycin resistance phenotype of KO-1304 arose from inability of mutated Xnr_2147 protein to repress XNR_2146. Introduction of additional copy of XNR_2146 into wild type strain increased antibiotic activity of the latter, attesting to the practical value of transporter genes for strain improvement. [ABSTRACT FROM AUTHOR]

Erysipelothrix rhusiopathiae is a bacterial pathogen that is the causative agent of erysipelas in a variety of animals, including swine, emus, turkeys, muskox, caribou, moose, and humans. This study aims to investigate the population structure and genomic features of Australian isolates of E. rhusiopathiae in the Australian pig industry and compare them to the broader scope of isolates worldwide. A total of 178 isolates (154 Australian, seven vaccine isolates, six international isolates, and 11 of unknown origin) in this study were screened against an MLST scheme and publicly available reference isolates, identifying 59 new alleles, with isolates separating into two main single locus variant groups. Investigation with BLASTn revealed the presence of the spaA gene in 171 (96%) of the isolates, with three main groups of SpaA protein sequences observed amongst the isolates. Novel SpaA protein sequences, categorised here as group 3 sequences, consisted of two sequence types forming separate clades to groups 1 and 2, with amino acid variants at positions 195 (D/A), 303 (G/E) and 323(P/L). In addition to the newly identified groups, five new variant positions were identified, 124 (S/N), 307 (Q/R), 323 (P/L), 379 (M/I), and 400 (V/I). Resistance screening identified genes related to lincomycin, streptomycin, erythromycin, and tetracycline resistance. Of the 29 isolates carrying these resistance genes, 82% belonged to SpaA group 2-N101S (n = 22) or 2-N101S-I257L (n = 2). In addition, 79% (n = 23) of these 29 isolates belonged to MLST group ST 5. Our results illustrate that Australia appears to have a unique diversity of E. rhusiopathiae isolates in pig production industries within the wider global context of isolates. [ABSTRACT FROM AUTHOR]

The South Saskatchewan River Basin (SSRB) is considered one of the most intensively farmed regions in Canada, with high densities of livestock and expansive areas of irrigated cropland. We measured concentrations of seven veterinary antimicrobials (VAs) in 114 surface water samples from feedlot environs and 219 samples from irrigation conveyances in the SSRB. Overall, detection frequencies in feedlot environs were 100% for chlortetracycline (CTC) and tetracycline (TC), 94% for monensin (MON), 84% for tylosin (TYL), 72% for lincomycin (LIN), 66% for erythromycin (ERY), and 23% for sulfamethazine (SMZ). For irrigation conveyances, detection frequencies for CTC and TC remained high (94–100%), but dropped to 18% for ERY, 15% for TYL, 10% for MON, and 4% for SMZ. Lincomycin was not detected in irrigation conveyance water. Maximum concentrations of VAs ranged from 1384 µg L−1 (TC) to 17 ng L−1 (SMZ) in feedlot environs while those in irrigation conveyances were 155 ng L−1 (TC) to 29 ng L−1 (ERY). High detection frequencies and median concentrations of VAs in both feedlot environs and irrigation conveyances were associated with high amounts of precipitation. However, an irrigation district (ID) with high livestock density (Lethbridge Northern) did not exhibit higher concentrations of VAs compared to IDs with less livestock, while levels of VAs in irrigation conveyances were less influenced by the degree of surface runoff. The ubiquity of CTC and TC in our study is likely a reflection of its widespread use in intensive livestock operations. Additional investigation is required to link environmental concentrations of VAs with livestock densities and increase our understanding of potential antimicrobial resistance in high-intensity agroecosystems. [ABSTRACT FROM AUTHOR]

This study aimed to investigate the presence and persistence of antibiotics in wastewater of four typical pharmaceutical manufactories in China and receiving water bodies and suggest the removal of antibiotics by the wastewater treatment process. It also evaluated the environmental impact of antibiotic residues through wastewater discharge into receiving water bodies. The results indicated that thirteen antibiotics were detected in wastewater samples with concentrations ranging from 57.03 to 726.79 ng/L. Fluoroquinolones and macrolides were the most abundant antibiotic classes found in wastewater samples, accounting for 42.5% and 38.7% of total antibiotic concentrations, respectively, followed by sulfonamides (16.4%) and tetracyclines (2.4%). Erythromycin-H2O, lincomycin, ofloxacin, and trimethoprim were the most frequently detected antibiotics; among these antibiotics, the concentration of ofloxacin was the highest in most wastewater samples. No significant difference was found in different treatment processes used to remove antibiotics in wastewater samples. More than 50% of antibiotics were not completely removed with a removal efficiency of less than 70%. The concentration of detected antibiotics in the receiving water bodies was an order of magnitude lower than that in the wastewater sample due to dilution. An environmental risk assessment showed that lincomycin and ofloxacin could pose a high risk at the concentrations detected in effluents and a medium risk in their receiving water bodies, highlighting a potential hazard to the health of the aquatic ecosystem. Overall, The investigation was aimed to determine and monitor the concentration of selected antibiotics in 4 typical PMFs and their receiving water bodies, and to study the removal of these substances in PMFs. This study will provide significant data and findings for future studies on antibiotics-related pollution control and management in water bodies. [ABSTRACT FROM AUTHOR]