Search Results (26)

Background: The therapeutic efficiency of cefquinome is currently limited by the low solubility and short half-life of its commercial sulfate form (SCFQ). This study aimed to improve these properties by preparing a novel crystalline cefquinome free acid (CFQ) via a reverse anti-solvent crystallization method. Methods: The optimal crystallization conditions were determined through a single factor test. And the product was characterized using X-ray powder diffraction, Fourier transform infrared spectroscopy, and scanning electron microscopy. Meanwhile, in vitro and in vivo pharmaceutical evaluation were conducted. Results: CFQ was sucessfully obtained and the optimal crystallization conditions were determined. Comparative in vitro studies showed that CFQ exhibited improved water solubility and dissolution rates compared to SCFQ. In vivo pharmacokinetic evaluations in chickens demonstrated that CFQ significantly prolonged the elimination half-life and increased the area under the concentration-time curve, achieving a relative bioavailability of 139.92%. Conclusions: The novel CFQ crystal effectively overcomes the crystallization difficulties of cefquinome and offers a promising alternative formulation with enhanced bioavailability and sustained drug action. Full article

Background/Objectives: Bovine colostrum, the initial milk produced by cows postpartum, contains an array of key nutritional, immune, and microbial components that support the calf’s physiological development, immune maturation, and intestinal colonization. The composition and quality of colostrum can be influenced by multiple factors, including seasonal variation, breed, parity, and farm management practices. This study investigated the microbial profile of Irish bovine colostrum and the influence of antibiotic therapy and parity. Methods: Bovine colostrum samples were collected from five Irish dairy farms that implemented different methods of dry cow therapy (DCT): natural or blanket. For blanket DCT, four of the five farms administered intramammary antibiotics at the start of the drying off period. Two farms administered a fourth-generation cephalosporin, cefquinome, and two farms used an antibiotic of the penicillin class, with the active ingredients consisting of procaine benzylpenicillin, penethamate hydriodide, and framycetin sulphate. One farm did not administer antibiotics but applied a teat sealant (natural DCT). Following calving, colostrum samples from 90 healthy dairy cows were analysed. Results: 16S rRNA sequence analysis revealed Firmicutes, Actinobacteriota, Bacteroidota, and Proteobacteria as the most abundant phyla across all treatment groups, with Acinetobacter, Corynebacterium, Facklamia, Jeotgalicoccus, Lactococcus, Leuconostoc, Psychrobacter, and Staphylococcus dominating at genus level. Parity did not significantly affect the microbial composition in this study, but antibiotic treatment did. Cows receiving no antibiotics showed distinct microbial clustering compared with antibiotic-treated cows (β-diversity, p < 0.001). Microbial diversity also differed between the antibiotic-treated groups, with significant changes in both α-diversity (p < 0.01) and β-diversity (p < 0.001), suggesting that the choice of antibiotic may also influence the microbiota. An influence of farm was also observed. Differential abundance analysis showed no increase in mastitis-associated genera in colostrum following natural DCT, although increased abundance was demonstrated with blanket DCT. Conclusions: Our findings substantiate the diverse and unique microbial composition of bovine colostrum. The data indicate that the microbial profile of bovine colostrum is influenced by antibiotic treatment administered during the dry period and affirms the latest policies inhibiting prophylactic antibiotic administration. Future studies should elucidate strain level changes in the colostrum microbiota following on-farm antibiotic use, assess the associated risks of antimicrobial resistance, and explore non-antibiotic alternatives for drying off cows. Evidently, the microbial composition of bovine colostrum is influenced by farm management strategies and optimizing these measures may further increase the valuable constituents of bovine colostrum and confer added health benefits to the new-born calf. Full article

To enhance the antibacterial activity of cefquinome (CEF) against Escherichia coli, a Carboxymethylcellulose sodium (CMCNa)/D-Mannosamine hydrochloride (DMH)-based nanogels delivery system capable of protecting CEF from gastric acid degradation while enabling intestinal sustained release and targeted antibacterial enhancement was developed. Systematic research was conducted on the best formulation, physicochemical characteristics, stability, gastrointestinal fluid-responsiveness, and antibacterial activity of the optimal formulation. The results showed that the optimized CEF nanogels demonstrated an enhanced loading capacity (13.0% ± 1.7%) and encapsulation efficiency (52.2% ± 1.0%). CEF nanogels appeared as uniform transparent spheres with a smooth surface under transmission electron microscopy and exhibited a three-dimensional porous network via scanning electron microscopy. More importantly, stability studies revealed that the CEF nanogels hold satisfactory stability. In addition, the formed CEF nanogels could effectively avoid the destruction of CEF by gastric acid in simulated gastric juice. In addition, they had the effect of slow and targeted release in the simulated intestinal tract. Compared to the free CEF, CEF nanogels have stronger antibacterial activity against Escherichia coli. In short, the prepared CEF nanogels had stronger antibacterial activity than CEF through sustained and targeted release. Full article

Bacteriological examination of uterine secretions provides essential information for the prevalence of bovine uterine pathogens and their influence on fertility. The objective of the present study was to determine the uterine pathogens in cases of clinical endometritis in two Holstein-Friesian dairy farms between 21 and 27 days after calving and the minimum inhibitory concentration (MIC) of 14 antimicrobials for Trueperella pyogenes (T. pyogenes). Overall, the prevalence of T. pyogenes (Farms A and B) was 46.3% and 22% (p < 0.01), respectively. In contrast, Farm B had significantly more cases (p < 0.001) of Escherichia coli, but the distribution of uterine pathogens was similar. Regarding the prevalence of any bacteria, Farm B also had significantly more bacteria (p < 0.001) than Farm A. T. pyogenes isolates were highly susceptible to amoxicillin, amoxicillin/clavulanic acid, tylosin, and cephalosporins, such as ceftiofur, cefquinome, and cephalexin with MIC₉₀ of ≤2 μg/mL. At the same time, MIC₉₀ of tulathromycin, lincomycin, and florfenicol were between 4 and 8 μg/mL and of doxycycline, enrofloxacin, oxytetracycline, and gentamicin, were between 16 and 32 μg/mL, respectively. Meanwhile, sulfamethoxazole/trimethoprim showed the highest MIC₉₀ (>32 μg/mL). In summary, T. pyogenes with high MIC₉₀ against oxytetracycline, gentamicin, and sulfamethoxazole/trimethoprim were found, which calls attention to the prudent use of antibiotics. Full article

Cefquinome is used to treat septicemia caused by Escherichia coli (E. coli) and respiratory infections caused by Streptococcus equi subsp. zooepidemicus in foals. However, studies reporting the use of cefquinome to target E. coli as pathogens of sepsis are lacking. Therefore, this study aimed to determine the optimal dosage regimen for cefquinome against E. coli using a PK/PD model. After the administration of 1 mg/kg cefquinome (intramuscularly or intravenously), blood samples were collected at different time points to determine the serum concentration of cefquinome via HPLC. The pharmacokinetic parameters were evaluated via NCA (WinNonlin 5.2.1 software). The main pharmacokinetic parameters of cefquinome in foals were as follows: after intravenous administration, the elimination half-life (T_1/2β) was 2.35 h, the area under the curve (AUC_0–last) was 12.33 μg·h/mL, the mean residence time (MRT_0–last) was 2.67 h, and the clearance rate (CL) was 0.09 L/h/kg. After intramuscular administration, the peak concentration (C_max) was 0.89 μg/mL, the time to reach the maximum serum concentration (T_max) was 2.16 h, T_1/2β was 4.16 h, AUC_0–last was 5.41 μg·h/mL, MRT_0–last was 4.92 h, CL was 0.15 L/h/kg, and the absolute bioavailability (F) was 43.86%. An inhibitory sigmoid Emax model was used to integrate the PK/PD indices with ex vivo antimicrobial effects to identify pharmacodynamic targets (PDTs). According to the dose calculation formula, the doses of intramuscularly administered cefquinome required to achieve bacteriostatic effects, bactericidal effects, and bactericidal elimination were 1.10, 1.66, and 2.28 mg/kg, respectively. However, further studies are warranted to verify the therapeutic efficacy of cefquinome in clinical settings. Full article

Cefquinome sulfate has a strong killing effect against Staphylococcus aureus (S. aureus), but bacterial resistance has become increasingly widespread. Experiments were conducted to investigate the pattern of adaptive resistance of S. aureus to cefquinome sulfate under different dosage regimens by using pharmacokinetic-pharmacodynamics (PK-PD) modeling, and the adaptive-resistant bacteria in different states were screened and subjected to transcriptomic sequencing. The results showed that the minimum inhibitory concentration of Staphylococcus aureus under the action of cefquinome sulfate was 0.5 μg/mL, the anti-mutation concentration was 1.6 μg/mL, and the mutation selection window range was 0.5~1.6 μg/mL. In the in vitro pharmacokinetic model to simulate different dosing regimens in the animal body, there are certain rules for the emergence of adaptive drug-resistant bacteria: the intensity of bacterial resistance gradually increased with culture time, and the order of emergence was tolerant bacteria (TO) followed by persistent bacteria (PE) and finally resistant bacteria (RE). The sequence reflected the evolution of adaptive drug resistance. Transcriptome Gene Ontology (GO) analysis revealed that differentially expressed genes were involved in cellular respiration, energy derivation by oxidation of organic compounds, and oxidation–reduction processes. The differentially expressed genes identified functioned in the synthesis of cell membranes, cytoplasm, and intracellular parts. A Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway analysis found that 65 genes were differentially expressed after cefquinome sulfate treatment, of which 35 genes were significantly upregulated and 30 genes were significantly downregulated. Five genes, sdhB, sdhA, pdhA, lpdA, and sucC, may be involved in network regulation. This study revealed the cross-regulation of multiple metabolic pathway networks and the targets of network regulation of S. aureus to produce adaptive drug resistance. The results will provide guidance for clinical drug use in animals infected with S. aureus. Full article

Collateral sensitivity is an evolutionary trade-off for bacteria where acquiring resistance to one antibiotic results in an increased sensitivity to another antibiotic. This study was designed to evaluate the collateral sensitivity of methicillin-resistant Staphylococcus aureus (MRSA) to β-lactam antibiotics induced by the evolution of resistance to apramycin. Collateral sensitivity to ampicillin, cephazolin, ceftriaxone, cefotaxime, cefepime and cefquinome occurred after MRSA were exposed to apramycin and induced to acquire resistance. This sensitivity was associated with reduced β-lactamase activity and decreased expression of the mecA gene. We also found a decrease in the proton motive force and decreased efflux activity. These results provide new insights into collateral sensitivity-based strategies for the treatment of MRSA. Full article

Antimicrobial resistance is an important health issue in human and veterinary medicine. The aim of this study was to monitor the antimicrobial resistance of three of the most important bacteria involved in porcine respiratory disease. A total of 465 isolates were tested during the 2017–2022 period for antimicrobial susceptibility for Actinobacillus pleuropneumoniae (n = 137), Streptococcus suis (n = 207), and Pasteurella multocida (n = 121) by disk diffusion method. The results were interpreted by CLSI breakpoints, where available. High rates of susceptibility (from 90 to >99%) were observed for cefquinome, ceftiofur, amoxicillin + clavulanic acid, amoxicillin, penicillin, ampicillin, florfenicol, enrofloxacin, marbofloxacin, and trimethoprim–sulfamethoxazole. A. pleuropneumoniae isolates showed high resistance to streptomycin (77%), gentamycin (45%), tilmicosin (39%) erythromycin (33%), oxytetracycline (19%), and tetracycline (18%). For S. suis, the highest rates of resistance were observed for streptomycin (98%), tetracycline (75%), oxytetracycline (72%), doxycycline (52%), and erythromycin (51%). P. multocida presented a high rate of resistance for streptomycin (63%), tilmicosin (29%), oxytetracycline (13%), and tetracycline (14%). Bacteria isolates maintained high susceptibility against antimicrobial agents usually used against the mainly respiratory tract pathogens of swine. Resistance for streptomycin, tetracycline, oxytetracycline, and tilmicosin was high for all the tested pathogens Full article

Porcine respiratory disease complex (PRDC) has been a major animal health, welfare, and economic problem in Hungary; therefore, great emphasis should be put on both the prevention and control of this complex disease. As antibacterial agents are effective tools for control, antibiotic susceptibility testing is indispensable for the proper implementation of antibacterial therapy and to prevent the spread of resistance. The best method for this is to determine the minimum inhibitory concentration (MIC) by the broth microdilution method. In our study, we measured the MIC values of 164 Actinobacillus pleuropneumoniae, 65 Pasteurella multocida, and 118 Streptococcus suis isolates isolated from clinical cases against the following antibacterial agents: amoxicillin, ceftiofur, cefquinome, oxytetracycline, doxycycline, tylosin, tilmicosin, tylvalosin, tulathromycin, lincomycin, tiamulin, florfenicol, colistin, enrofloxacin, and sulfamethoxazole-trimethoprim. Outstanding efficacy against A. pleuropneumoniae isolates was observed with ceftiofur (100%) and tulathromycin (100%), while high levels of resistance were observed against cefquinome (92.7%) and sulfamethoxazole-trimethoprim (90.8%). Ceftiofur (98.4%), enrofloxacin (100%), florfenicol (100%), and tulathromycin (100%) were found to be highly effective against P. multocida isolates, while 100% resistance was detected against the sulfamethoxazole-trimethoprim combination. For the S. suis isolates, only ceftiofur (100%) was not found to be resistant, while the highest rate of resistance was observed against the sulfamethoxazole-trimethoprim combination (94.3%). An increasing number of studies report multi-resistant strains of all three pathogens, making their monitoring a high priority for animal and public health. Full article

Mannheimia haemolytica is the main pathogen contributing to pneumonic pasteurellosis in sheep. The aim of this study was to investigate the antimicrobial resistance levels in M. haemolytica isolates from the lungs of slaughtered sheep and to examine the genetic resistance mechanisms involved. A total of 256 M. haemolytica isolates, 169 from lungs with pneumonic lesions and 87 from lungs without lesions, were analyzed by the disk diffusion method for 12 antimicrobials, and the whole genome of 14 isolates was sequenced to identify antimicrobial resistance determinants. Levels of phenotypic resistance ranged from <2% for 10 antimicrobials (amoxicillin, amoxicillin-clavulanic, ceftiofur, cefquinome, lincomycin/spectinomycin, gentamicin, erythromycin, florfenicol, enrofloxacin, and doxycycline) to 4.3% for tetracycline and 89.1% for tylosin. Six isolates carried tetH genes and four isolates carried, in addition, the strA and sul2 genes in putative plasmid sequences. No mutations associated with macrolide resistance were identified in 23 rDNA sequences, suggesting that the M. haemolytica phenotypic results for tylosin should be interpreted with care in the absence of well-established epidemiological and clinical breakpoints. The identification of strains phenotypically resistant to tetracycline and of several resistance genes, some of which were present in plasmids, highlights the need for continuous monitoring of susceptibility patterns in Pasteurellaceae isolates from livestock. Full article

The development of drug-resistance in the opportunistic pathogen Escherichia coli has become a global public health concern. Due to the share of similar flora between pets and their owners, the detection of pet-origin antibiotic-resistant E. coli is necessary. This study aimed to detect the prevalence of feline-origin ESBL E. coli in China and to explore the resistance elimination effect of garlic oil to cefquinome on ESBL E. coli. Cat fecal samples were collected from animal hospitals. The E. coli isolates were separated and purified by indicator media and polymerase chain reaction (PCR). ESBL genes were detected by PCR and Sanger sequencing. The MICs were determined. The synergistic effect of garlic oil and cefquinome against ESBL E. coli was investigated by checkerboard assays, time-kill and growth curves, drug-resistance curves, PI and NPN staining, and a scanning electronic microscope. A total of 80 E. coli strains were isolated from 101 fecal samples. The rate of ESBL E. coli was 52.5% (42/80). The prevailing ESBL genotypes in China were CTX-M-1, CTX-M-14, and TEM-116. In ESBL E. coli, garlic oil increased the susceptibility to cefquinome with FICIs from 0.2 to 0.7 and enhanced the killing effect of cefquinome with membrane destruction. Resistance to cefquinome decreased with treatment of garlic oil after 15 generations. Our study indicates that ESBL E. coli has been detected in cats kept as pets. The sensitivity of ESBL E. coli to cefquinome was enhanced by garlic oil, indicating that garlic oil may be a potential antibiotic enhancer. Full article

(1) Background: The resistance levels of Escherichia coli, Salmonella spp., Pseudomonas spp., Staphylococcus spp., etc., isolated from the nasal cavity and the rectum of Dama dama deer from three hunting grounds in Western Romania were assessed. (2) Methods: The analysis was completed using the diffusimetric method, compliant with CLSI reference standards, and with Vitek-2 (BioMérieux, France), on 240 samples. (3) Results: The results were statistically analyzed (by one-way ANOVA) revealing that in four of the ten E. coli strains isolated from animals, 87.5% (p < 0.001) resistance was found. E. coli strains were resistant to cephalexin (100%); seven strains were resistant to cephalothin and ampicillin; six were resistant to cefquinome and cefoperazone; five were resistant to amoxicillin/clavulanic acid; and four were resistant to ceftiofur. However, E. coli was sensitive to amikacin (100%). The most efficient structures were beta-lactams, amikacin, and imipenem, to which all 47 strains studied (100%) were sensitive, followed by nitrofurantoin, to which 45 strains (95.7%) were sensitive, neomycin, to which 44 strains (93.6%) were sensitive, ceftiofur, to which 43 strains (91.5%) were sensitive, and trimethoprim/sulfamethoxazole and marbofloxacin, to which 42 strains (89.4%) were sensitive. (4) Conclusions: In wild animal populations, where a human presence is frequently reported, including a constant presence of domestic animals, despite the perceived low risk of emerging resistance to antimicrobials, resistance is likely to develop frequently. Full article

As set in the maximum residue limit regulations of the European Commission, this study aimed to obtain the residual parameters in milk with optimized UPLC-MS/MS conditions and to determine the conclusive drug withdrawal period to ensure food safety. In this research, an ultra-high performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) method was developed to study cefquinome sulfate’s residue elimination in milk and to calculate cefquinome’s withdrawal period. Twelve healthy cows free of endometritis were selected for the experiment. Before using the drug, the vaginal orifice and perineum of each cow was disinfected. One dose of intrauterine perfusion was used for each cow, followed by an additional dose after 72 h. Before administration and 12 h, 18 h, 24 h, 36 h, 42 h, 48 h, 60 h, 66 h, 72 h, 84 h, 90 h, and 96 h after the last dose, milk (10 mL) was gathered from each cow’s teat and pooled. For the measurement of cefquinome concentrations in milk, UPLC-MS/MS was performed. A calibration curve was generated using linear regression as follows: Y = 250.86X − 102.29, with a correlation coefficient of 0.9996; the limits of detection and the limits of quantitation were 0.1 μg·kg⁻¹ and 0.2 μg·kg⁻¹, respectively. The average recovery of cefquinome was 88.60 ± 16.33% at 0.2 μg·kg⁻¹, 100.95 ± 2.54% at 10 μg·kg⁻¹, and 97.29 ± 1.77% at 50 μg·kg⁻¹. For 5 consecutive days at the three spiking levels, the intra and inter-day relative standard deviations (RSD) were 1.28%–13.73% and 1.81%–18.44%, respectively; the residual amount of cefquinome was less than the maximum residue limit of 20 μg·kg⁻¹, 36 h after administration; and the residual amount was less than the limit of detection (0.1 μg·kg⁻¹) 48 h after administration. The withdrawal time of cefquinome in cow’s milk was 39.8 h, as calculated using WTM1.4 software. In terms of clinical practical use, the withdrawal period of milk was temporarily set at 48 h after the administration of the cefquinome sulfate uterus injection to cows, in accordance with the recommended dose and course. Full article

Globally, cephalosporin therapy failure is a serious problem for infection control. One causative agent of cephalosporin-resistant infections is multidrug-resistant (MDR) E. coli producing extended-spectrum β-lactamases (ESBLs) and/or plasmid-encoded AmpC (pAmpC) β-lactamases. We evaluated the occurrence of ESBL/pAmpC genetic determinants in phenotypically MDR E. coli isolated from clinical samples of blood, faeces, ear effusion, urine and sputum from a UK hospital. Phenotypic resistance profiling for 18 antibiotics (from seven classes) showed that 32/35 isolates were MDR, with resistance to 4–16 of the tested antibiotics. Of the isolates, 97.1% showed resistance to ampicillin, 71.4% showed resistance to co-amoxiclav, cefotaxime, ceftazidime and ceftiofur, and 68.5% showed resistance to cefquinome. bla_CTX-M, bla_TEM and bla_OXA-1 genes were detected in 23, 13 and 12 strains, respectively, and Intl1 was detected in 17 isolates. The most common subtypes among the definite sequence types were CTX-M-15 (40%) and TEM-1 (75%). No E. coli isolates carried pAmpC genes. Significant correlations were seen between CTX-M carriage and cefotaxime, ceftiofur, aztreonam, ceftazidime and cefquinome resistance; between bla_CTX-M, bla_TEM and bla_OXA-1 carriage and ciprofloxacin resistance; and between Intl1 carriage and trimethoprim/sulfamethoxazole resistance. Thus, MDR phenotypes may be conferred by a relatively small number of genes. The level and pattern of antibiotic resistance highlight the need for better antibiotic therapy guidelines, including reduced use and improved surveillance. Full article

The presence of antibiotics in milk is a significant problem affecting the technological safety of dairy products. The aim of the study was to determine the sensitivity of yoghurt cultures to residual levels of selected cephalosporin antibiotics (cephalexin, cefoperazone, cefquinome, cefazolin, and ceftiofur). Five yoghurt cultures were selected containing strains of Lactobacillus delbrueckii subsp. bulgaricus and Streptococcus thermophilus. Artificially fortified milk samples (whole pasteurized milk; 85 °C; 3–5 s) with cephalosporins at a concentration of the maximum residue limit were used to evaluate the sensitivity of the yoghurt cultures by monitoring the pH, titratable acidity, and the concentration of selected organic acids (lactic, pyruvic, citric, acetic, orotic, oxalic, formic, uric, and succinic acids) at the end of fermentation (43 °C; 4–5.5 h; pH ≤ 4.6). The titratable acidity was determined by the Soxhlet–Henkel method and the organic acid concentration was monitored by reversed-phase HPLC. Ceftiofur had the greatest effect on the yoghurt culture activity, with a statistically highly significant effect (p < 0.05) on the pH, titratable acidity, and the content of lactic, pyruvic, and acetic acids in all cultures. Other cephalosporins also showed an inhibitory effect on yoghurt metabolism as seen by the evaluation of the lactic and pyruvic acid concentrations. Full article