Compound organic acid could improve the growth performance, immunity and antioxidant properties, and intestinal health by altering the microbiota profile of weaned piglets.

This study aimed to investigate the impact of compound organic acid (COA) and chlortetracycline (CTC) on serum biochemical parameters, intestinal health, and growth performance of weaned piglets. Twenty-four piglets (24 d of age) were randomly allocated into three treatments with eight replicate pens (one piglet per pen). Feed the basal diet or a diet containing 3,000 mg/kg COA or 75 mg/kg CTC, respectively. Results showed that both COA and CTC significantly increased average daily gain and reduced diarrhea rates (P < 0.05). They also upregulated serum total antioxidant capacity and downregulated serum interleukin (IL-10) levels (P < 0.05), increased crude protein digestibility and propionic acid concentration in the colon, and decreased spermidine and putrescine contents (P < 0.05). Intestinal microbiota analysis revealed that both COA and CTC increased the Shannon and Chao1 index and decreased the relative abundance of Blautia and Roseburia, but increased the relative abundance of Clostridium-sensu-stricto-1. Correlation analysis indicated that Clostridium-sensu-stricto-1 may be closely related to inflammation levels and microbial metabolites in piglets. Based on the results, COA may be a potential substitute for CTC to reduce antibiotic use and biogenic amine emission while improving piglet growth and intestinal health.

Weaned piglets face challenges due to their underdeveloped digestive system, resulting in high gastrointestinal tract pH and insufficient enzyme secretion. To address this issue, we found supplementing piglet diets with 3,000 mg/kg of compound organic acid positively impacted the immune and antioxidant levels of piglets, promoted their intestinal health, improved nutrient digestibility, and enhanced their overall growth performance. These benefits were attributed to the regulation of intestinal microbiota by the compound organic acid. By improving piglet health and growth, this research offers a potential solution to the challenges of piglet weaning stress.

Adsorption and transformation of tetracyclines on alpha alumina particles with surface modification by anionic surfactant.

The adsorption and transformation of tetracyclines (TCs) antibiotics, including oxytetracycline (OTC), chlortetracycline (CTC), and tetracycline (TC), on the sodium dodecyl sulfate (SDS) surfactant-modified α-Al2O3 particles were comprehensively investigated in this study. The TCs adsorption was significantly enhanced by using the modified adsorbents compared with the use of the unmodified adsorbents. The experimental conditions were systematically optimized and found to be pH 4, NaCl 1 mM, the contact time of 180 min, and the adsorbent dosage of 25 mg. mL-1. The high maximum adsorption capacities were approximately 320, 85, and 91 mg. g-1 for TC, OTC, and CTC, respectively. Meanwhile, the great removal efficiencies of the three antibiotics TC, OTC, and CTC were correspondingly 91.85, 88.4, and 98.3%. The TCs adsorption isotherm and kinetics on the SDS-modified α-Al2O3 particles mainly governed by the electrostatic and hydrophobic interactions were clarified by a suitable two-step model, the Fourier transform infrared spectroscopy (FT-IR) and zeta potential measurements. Meanwhile, the TCs structural transformation determined by the liquid chromatography with tandem mass spectrometry (LC-MS/MS) measurement was promoted through the adsorption on the α-Al2O3 surface. The TCs transformation rates strongly affected by the TCs adsorption were in the order of CTC > TC > OTC. The found results are promised that the SDS-modified α-Al2O3 particles might behave as high-performance adsorbents to remove the TCs from aqueous solutions.

Enhanced chlortetracycline removal by iron oxide modified spent coffee grounds biochar and persulfate system.

Chlortetracycline (CTC) is a tetracycline derivative antibiotic that has been widely used in the livestock industry for prophylactic and therapeutic purposes. Effective measures should be taken to decrease the environmental risks associated with CTC-rich waste. Biochar produced by biomass waste showed great potential for organic contaminants removal by adsorption and catalytic degradation. This study prepared iron oxide-modified coffee grounds biochar (CGF) at different temperatures for enhanced CTC removal by adsorption and degradation. The main mechanism for CTC removal was found to be electrostatic interaction. In addition, pore diffusion, hydrogen bonds, and π-π bonds also contributed to CTC adsorption. Maximum CTC adsorption capacity was 223.63 mg/g for CGF800 (CGF prepared at 800 °C pyrolysis). The free radical content of CGF600 (CFG prepared at 600 °C pyrolysis) was higher than CGF800, and there were no significant advantages in using biochar prepared at a higher temperature for persulfate activation. The ion mass-to-charge ratio (M/z) is used to describe the ratio of mass to charge of an ion or peak, which can infer compound structure. The structure of CTC degradation products was analyzed by UPLC-MS, and the M/z values were determined as 444, 273, and 154. Thus, pyrolysis of coffee grounds at higher temperatures increased CTC adsorption capacity, and CGF can indirectly assist in CTC degradation by persulfate activation.

Antibiotic Chlortetracycline Causes Transgenerational Immunosuppression via NF-κB.

The extensive and increasing global use of antibiotics results in the ubiquitous presence of antibiotics in the environment, which has made them "pseudo persistent organic contaminants." Despite numerous studies showing wide adverse effects of antibiotics on organisms, the chronic environmental risk of their exposure is unknown, and the molecular and cellular mechanisms of antibiotic toxicity remain unclear. Here, we systematically quantified transgenerational immune disturbances after chronic parental exposure to environmental levels of a common antibiotic, chlortetracycline (CTC), using zebrafish as a model. CTC strongly reduced the antibacterial activities of fish offspring by transgenerational immunosuppression. Both innate and adaptive immunities of the offspring were suppressed, showing significant perturbation of macrophages and neutrophils, expression of immune-related genes, and other immune functions. Moreover, these CTC-induced immune effects were either prevented or alleviated by the supplementation with PDTC, an antagonist of nuclear factor-κB (NF-κB), uncovering a seminal role of NF-κB in CTC immunotoxicity. Our results provide the evidence in fish that CTC at environmentally relevant concentrations can be transmitted over multiple generations and weaken the immune defense of offspring, raising concerns on the population hazards and ecological risk of antibiotics in the natural environment.

Removal of chlortetracycline from water using spent tea leaves-based biochar as adsorption-enhanced persulfate activator.

Antibiotic compounds have caused serious environmental concerns. In this study, we developed an effective technology for treatment of chlortetracycline (CTC), a widely used antibiotic compound. A natural heteroatom-doped spent tea leaves-based biochar (STLB) with excellent adsorption and catalytic property was prepared by simple thermal treatment. An adsorption-promoted persulfate-based advanced oxidation process (PS-AOP) using STLB was studied for CTC removal. The results showed that the as-prepared STLB presented favorable adsorption affinity towards CTC with the maximum adsorption capacity of 627 mg g-1. Meanwhile, CTC enriched on the surface of STLB was good for in-situ decomposition of CTC and nearly 97.4 % of CTC was removed within 30 min of pre-adsorption and 60 min of subsequent degradation. The STLB had excellent recyclability and wide pH tolerance range of 3.0-9.0 in combined pre-adsorption and PS-AOP. Reactive oxygen species analysis confirmed that CTC degradation was mainly due to non-radical (singlet oxygen, 1O2) and radicals (SO4- and OH). This study suggests that STLB is a promising adsorption-enhanced PS activator for the treatment of refractory wastewater and also provides a strategy of waste control by spent tea leaves.

Effects of chlortetracycline on growth performance and intestinal functions in weaned piglets.

AIM: Weaning stress can cause serious damage to piglet's health. Chlortetracycline (CTC) is widely used to ameliorate weaning stress and prevent infectious diseases in weaned piglets. However, antibiotics as growth promoters have to be limited because of increased antimicrobial resistance. In this study, we evaluated the effects of CTC on growth performance and intestinal functions in order to provide evidence for seeking antibiotic substitutes in weaned piglets. METHODS AND RESULTS: A total of 20 weaned piglets were fed a basal diet or a diet supplemented with 75 mg/kg CTC. CTC decreased the crypt depth and increased the ratio of villus height to crypt depth, whilst failing to affect growth performance and serum biochemical parameters and cytokines. 16S rRNA sequencing suggested that CTC supplementation had no effect on the diversity and composition of colonic microbiota. CONCLUSION: We speculated that gut microbiota is no longer sensitive to a low concentration of CTC due to the long-term use and low bioavailability of CTC in weaned piglets.

Chlortetracycline alters microbiota of gut or faeces in pigs and leads to accumulation and migration of antibiotic resistance genes.

In this study, we investigated the effect of long-term use of chlortetracycline (CTC) on the gut microbiota composition and metabolism profiles in pigs, and the variation of antibiotic resistance genes (ARGs) and microbial communities in faeces and manure during aerobic composting (AC) and anaerobic digestion (AD). The pigs were fed the same basal diet supplemented with or without 75 mg/kg CTC, and fresh faeces of 30-, 60-, 90-, and 120-day-old pigs were collected from the CTC group. The results showed that CTC reduced the diversity of the gut microbiota significantly and changed its structure. Metabolomics analysis of intestinal contents revealed 23 differentially abundant metabolites, mainly organic acids, carbohydrates, and amino acids. Metabolic pathways, such as the TCA cycle, propionate metabolism, and pyruvate metabolism, were changed. From 30 to 120 days of age, the amount of CTC residues in faeces and the abundance of 3 tetracycline resistance genes increased significantly, and it was positively correlated with tetC, tetG, tetW, sul1 and intI2. CTC residue levels and ARGs abundance gradually decreased with fermentation time, and AC was better than AD at reducing ARGs abundance. The results suggest that in-feed CTC can reduce the diversity of the gut microbiota, change the structure, function and metabolism of the bacterial community, and increase the abundance of ARGs in faeces.

Soybean oligosaccharides attenuate odour compounds in excreta by modulating the caecal microbiota in broilers.

Abatement of odour emissions in poultry production is important to ensure the quality and safety in the poultry industry as well as for the benefit of the environment. This study was conducted to compare the effects of supplementation with different amounts of dietary soybean oligosaccharides (SBO) and chlortetracycline (CHL) on the major odour-causing compounds in the excreta and on the caecal microbiota in broiler chickens. One-day-old broiler chickens were subjected to a 42-day experiment involving 6 treatments with 6 replicates per treatment (10 birds/cage). The treatments were as follows: negative control (NC; basal diet), positive control (PC; basal diet supplemented with CHL) and basal diet supplemented with 0.5, 2.0, 3.5 and 5.0 g/kg SBO (0.5SBO, 2.0SBO, 3.5SBO and 5.0SBO, respectively). Fresh excreta were sampled for the analysis of odour compounds by HPLC. Caecum content was collected for the caecal microbiota analysis through 16S rRNA sequencing. Results showed that on day 42, the excreta indole concentration in the broilers fed with 2.0SBO, 3.5SBO and 5.0SBO and PC was significantly decreased (P < 0.01) compared with that in the NC-fed broilers. The excreta skatole concentration (P < 0.001) and pH (P < 0.05) were also decreased by SBO and CHL, and they were lowest in birds fed with 3.5SBO. The formate concentrations in birds fed with 3.5SBO and 5.0SBO were higher than those in birds fed with the other diets (P < 0.001). Similarly, acetate concentration (P = 0.003) was increased in birds fed with 3.5SBO. Deep sequencing of 16S rRNA revealed that the composition of the caecal microbial digesta was slightly or significantly changed by the SBO or by the CHL supplementation, respectively. The SBO supplementation decreased the abundance of Bacteroides, Bilophilaand Escherichia, which were associated with the skatole and indole concentrations in the excreta. By contrast, the CHL supplementation demonstrated a strong tendency to enrich Ruminococcus and to reduce the abundance of Rikenella. These results indicated that supplementation with dietary SBO is beneficial in attenuating the concentration of odour-causing compounds and that it modulates the composition of caecal microbiota in broiler chickens.

Determination of plasma-chlortetracycline (CTC) concentrations in grazing beef cattle fed one of four FDA approved free-choice CTC-medicated minerals.

Control of active bovine anaplasmosis in the United States is predicated on the use of chlortetracycline (CTC)-medicated feed throughout the vector season. However, data describing population pharmacokinetics of chlortetracycline in cows, on pasture, having free-choice access to CTC-medicated mineral for consecutive months is lacking. This study documented plasma-CTC concentrations in grazing cows during peak vector season in an anaplasmosis endemic herd. Each pasture was administered one of the four Food and Drug Administration approved CTC-medicated mineral formulations and were assigned as follows: 0.77 g/kg, Aureo Anaplaz C700 Pressed (Sweetlix Livestock Supplements, Mankato, MN); 5.5 g/kg, Purina Anaplasmosis Block (Purina Animal Nutrition, Gray Summit, MO); 6.6 g/kg, Stockmaster Aureo FC C6000 Mineral (Hubbard Feeds, Mankato, MN); 8.8 g/kg, MoorMan's Special Range Minerals AU 168XFE (ADM Animal Nutrition, Quincy, IL). Blood samples were collected monthly for determining plasma drug concentration by Ultra performance liquid chromatography (UPLC) and mass spectrometry. Continued plasma-CTC monitoring allowed for characterization of trends between treatment groups (pastures), age groups (<3 yr or >4 yr), and sampling times (June to October). Results indicate formulation (pasture) and time were significant factors affecting concentrations of CTC in plasma. Cows exposed to 5.5 g/kg block formulation recorded higher CTC plasma concentrations compared with other pasture groups (P = 0.037). Plasma-CTC concentrations increased over time (month of measurement; P = 0.0005). Specifically, concentrations measured after 5 months of continuous CTC treatment were higher than those measured in earlier months.

Exploring microbial community structure and biological function in manured soil during ten repeated treatments with chlortetracycline and ciprofloxacin.

The changes of enzyme activities, microbial community structure and function, and the diversity and resistance level of antibiotic-resistant bacteria (ARB) were studied in soil during ten repeated treatments with chlortetracycline (CTC) and/or ciprofloxacin (CIP) together with organic manure (OM) under laboratory conditions. The activities of neutral phosphatase (NPA) and catalase (CAT) displayed the suppression-recovery-stimulation trend in the OM&CTC treatment but the stimulation trend in the OM&CTC&CIP treatment. The NPA was stimulated but the CAT was little affected in the OM&CIP treatment. Soil microbial functional diversity displayed the suppression-recovery-stimulation trend in the OM&CTC and OM&CTC&CIP treatments and the stimulation-suppression trend in the OM&CIP treatment with the treatment frequency. Metagenomic analysis showed that the relative abundances of Actinobacteria and Firmicutes in the antibiotic treatment significantly increased by 0.5-235.6%, but that of Proteobacteria decreased by 0.2-27.3% compared to the control with the treatment frequency. Furthermore, the relative abundances of dominant bacterial genera including Streptomyces, Actinomadura, Mycobacterium, and Streptococcus in the antibiotic treatment significantly increased by 1.1-10433.3% compared to the control. Meanwhile, repeated antibiotic treatments induced a significant increase in the diversity and resistance level of ARB isolates, especially in the OM&CTC treatment. It is concluded that repeated treatments with CTC and/or CIP can alter enzyme activities, microbial community structure and function, and increase the diversity and resistance level of ARB isolates.

The chicken gut metagenome and the modulatory effects of plant-derived benzylisoquinoline alkaloids.

BACKGROUND: Sub-therapeutic antibiotics are widely used as growth promoters in the poultry industry; however, the resulting antibiotic resistance threatens public health. A plant-derived growth promoter, Macleaya cordata extract (MCE), with effective ingredients of benzylisoquinoline alkaloids, is a potential alternative to antibiotic growth promoters. Altered intestinal microbiota play important roles in growth promotion, but the underlying mechanism remains unknown. RESULTS: We generated 1.64 terabases of metagenomic data from 495 chicken intestinal digesta samples and constructed a comprehensive chicken gut microbial gene catalog (9.04 million genes), which is also the first gene catalog of an animal's gut microbiome that covers all intestinal compartments. Then, we identified the distinctive characteristics and temporal changes in the foregut and hindgut microbiota. Next, we assessed the impact of MCE on chickens and gut microbiota. Chickens fed with MCE had improved growth performance, and major microbial changes were confined to the foregut, with the predominant role of Lactobacillus being enhanced, and the amino acids, vitamins, and secondary bile acids biosynthesis pathways being upregulated, but lacked the accumulation of antibiotic-resistance genes. In comparison, treatment with chlortetracycline similarly enriched some biosynthesis pathways of nutrients in the foregut microbiota, but elicited an increase in antibiotic-producing bacteria and antibiotic-resistance genes. CONCLUSION: The reference gene catalog of the chicken gut microbiome is an important supplement to animal gut metagenomes. Metagenomic analysis provides insights into the growth-promoting mechanism of MCE, and underscored the importance of utilizing safe and effective growth promoters.

Macleaya cordata helps improve the growth-promoting effect of chlortetracycline on broiler chickens.

Chlortetracycline (CTC), one kind of common antibiotic for prevention and treatment of various diseases, also exhibits good performance in accelerating the growth of livestock. Macleaya cordata, a traditional Chinese medicine, is usually used as a natural additive in livestock because of its anti-microbial, anti-fungal, anti-inflammatory, and pesticidal activity. In this work, we studied whether M. cordata helps regulate the growth-promoting effect of CTC on broiler chickens. It is demonstrated that M. cordata improves the growth-promoting effect of CTC on growth performance indices of broiler chickens, such as survival rate, daily weight, and feed to weight rate. M. cordata also delays the maximum of CTC residues in plasma. It may depend on the higher values of operational taxonomic unit (OTU) and the indices of α diversity driven by simultaneous use of CTC and M. cordata.

Illumina Miseq platform analysis caecum bacterial communities of rex rabbits fed with different antibiotics.

Antibiotics have been widely used for the prevention and the treatment of diseases to humans and animals, and they have fed additives for agricultural animals to promote growth. However, there is a growing concern over the practice due to its side effects on intestinal microbial communities which plays a vital role in animals' health. To investigate the effect of antibiotics on the bacterial population of the caecum in rex rabbits, 80 rex rabbits were randomly divided into four groups: control group (B, basal diet), chlortetracycline group (C, 50 mg/kg), colistin sulfate group (S, 20 mg/kg) and zinc bacitracin group (Z, 40 mg/kg). Caecum microbial communities of rex rabbits from the four groups were analyzed through Illumina Miseq platform after being fed 28 days. The results showed that most obtained sequences belongs to Firmicutes followed by Bacteroidetes, and the ratio of Bacteroidetes/Firmicutes in C group (42.31 %) was higher than that in Z group (21.84 %). Zinc bacitracin supplementation caused a significant decreased of the Proteobacteria phylum and Lactobacillus spp. (P < 0.05), while the Lactobacillus spp. significantly increased in S group (P < 0.05). In addition, Ruminococcus spp., especially Ruminococcus albus were the predominant bacterial species found in both S and Z groups. The proportion of Coprococcus spp. significantly increased in Z group (P < 0.05). These findings suggested that the antibiotics used may cause significant changes in the caecum microbiota of rex rabbits, and we also found C group had a similarity caecum bacteria structure with B group which was probably due to the high levels of chlortetracycline resistance.

Effects of chlortetracycline and copper supplementation on the prevalence, distribution, and quantity of antimicrobial resistance genes in the fecal metagenome of weaned pigs.

Use of in-feed antibiotics such as chlortetracycline (CTC) in food animals is fiercely debated as a cause of antimicrobial resistance in human pathogens; as a result, alternatives to antibiotics such as heavy metals have been proposed. We used a total community DNA approach to experimentally investigate the effects of CTC and copper supplementation on the presence and quantity of antimicrobial resistance elements in the gut microbial ecology of pigs. Total community DNA was extracted from 569 fecal samples collected weekly over a 6-week period from groups of 5 pigs housed in 32 pens that were randomized to receive either control, CTC, copper, or copper plus CTC regimens. Qualitative and quantitative PCR were used to detect the presence of 14 tetracycline resistance (tet) genes and to quantify gene copies of tetA, tetB, blaCMY-2 (a 3rd generation cephalosporin resistance gene), and pcoD (a copper resistance gene), respectively. The detection of tetA and tetB decreased over the subsequent sampling periods, whereas the prevalence of tetC and tetP increased. CTC and copper plus CTC supplementation increased both the prevalence and gene copy numbers of tetA, while decreasing both the prevalence and gene copies of tetB. In summary, tet gene presence was initially very diverse in the gut bacterial community of weaned pigs; thereafter, copper and CTC supplementation differentially impacted the prevalence and quantity of the various tetracycline, ceftiofur and copper resistance genes resulting in a less diverse gene population.

Effects of chlortetracycline and copper supplementation on antimicrobial resistance of fecal Escherichia coli from weaned pigs.

Feed-grade chlortetracycline (CTC) and copper are both widely utilized in U.S. pig production. Cluster randomized experiment was conducted to evaluate the effects of CTC and copper supplementation in weaned pigs on antimicrobial resistance (AMR) among fecal Escherichia coli. Four treatment groups: control, copper, CTC, or copper plus CTC were randomly allocated to 32 pens with five pigs per pen. Fecal samples were collected weekly from three pigs per pen for six weeks. Two E. coli isolates per fecal sample were tested for phenotypic and genotypic resistance against antibiotics and copper. Data were analyzed with multilevel mixed effects logistic regression, multivariate probit analysis and discrete time survival analysis. CTC-supplementation was significantly (99% [95% CI=98-100%]) associated with increased tetracycline resistance compared to the control group (95% [95% CI=94-97%]). Copper supplementation was associated with decreased resistance to most of the antibiotics tested, including cephalosporins, over the treatment period. Overall, 91% of the E. coli isolates were multidrug resistant (MDR) (resistant to ≥3 antimicrobial classes). tetA and blaCMY-2 genes were positively associated (P<0.05) with MDR categorization, while tetB and pcoD were negatively associated with MDR. tetA and blaCMY-2 were positively associated with each other and in turn, these were negatively associated with both tetB and pcoD genes; which were also positively associated with one another. Copper minimum inhibitory concentration was not affected by copper supplementation or by pcoD gene carriage. CTC supplementation was significantly associated with increased susceptibilities of E. coli to copper (HR=7 [95% CI=2.5-19.5]) during treatment period. In conclusion, E. coli isolates from the nursery pigs exhibited high levels of antibiotic resistance, with diverse multi-resistant phenotypic profiles. The roles of copper supplementation in pig production, and pco-mediated copper resistance among E. coli in particular, need to be further explored since a strong negative association of pco with both tetA and blaCMY-2 points to opportunities for selecting a more innocuous resistance profile.