The green turtle (Chelonia mydas) is listed as a globally endangered species and is vulnerable to anthropogenic threats, including environmental pollution. This study investigated the antimicrobial resistance of Gram-negative bacteria isolated from wild green turtles admitted to a sea turtle rehabilitation center in Taiwan. For this investigation, cloacal and nasal swab samples were collected from 28 green turtles between 2018 and 2020, from which a total of 47 Gram-negative bacterial isolates were identified. Among these, Vibrio spp. were the most dominant isolate (31.91%), and 89.36% of the 47 isolates showed resistance to at least one of 18 antimicrobial agents tested. Isolates resistant to one (6.38%), two (8.51%), and multiple (74.47%) antimicrobials were observed. The antimicrobial agents to which isolates showed the greatest resistance were penicillin (74.47%), followed by spiramycin, amoxicillin, and cephalexin. The antimicrobial-resistance profiles identified in this study provide useful information for the clinical treatment of sea turtles in rehabilitation facilities. The results of our study also imply that wild green turtles may be exposed to polluting effluents containing antimicrobials when the turtles traverse migratory corridors or forage in feeding habitats. To benefit sea turtle conservation, future research should focus on (1) how to prevent pollution from antimicrobials in major green turtle activity areas and (2) identifying sources of antimicrobial-resistant bacterial strains in coastal waters of Taiwan.

Spiramycin is a widely used macrolide antibiotic and exists at high concentration in production wastewater. A thermal-acid hydrolytic pretreatment using silicotungstic acid (STA) under microwave (MW) irradiation was suggested to mitigate spiramycin from production wastewater. Positive correlations were observed between STA dosage, MW power, interaction time and the hydrolytic removal efficiencies, and an integrative equation was generalized quantitively. Rapid and complete removal 100 mg/L of spiramycin was achieved after 8 min of reaction with 1.0 g/L of STA under 200 W of MW irradiation, comparing to 30.1% by MW irradiation or 15.9% by STA alone. The synergetic effects of STA and MW irradiation were originated from the dissociated-proton catalysis by STA and the dipolar rotation heating effect of MW. STA performed much better than the mineral acid H2SO4 under MW, due to the much stronger Brönsted acidity and higher Hammett acidity. After 8 min, 98.0% of antibacterial potency was also reduced. The m/z 558.8614 fragment (P1) and m/z 448.1323 fragment (P2) were identified as the primary products, which were formed by breaking glucosidic bonds and losing mycarose and forosamine for P1 and further mycaminose moiety for P2. Finally, production wastewater with 433 mg/L of spiramycin was effectively treated using this thermal-acid hydrolytic method. Spiramycin and its antibacterial potency both dropped to 0 after 6 min. The potency drop was supposed from the losing of mycarose and/or forosamine. To decrease both the concentration of spiramycin and its antibacterial potency, combinedly using STA and MW was suggested in this work to break down the structural bonds of the functional groups rather than to destroy the whole antibiotic molecules. It is promising for pretreating spiramycin-contained production wastewater to mitigate both the antibiotic and its antibacterial potency.

Nitrogen significantly regulated (p < 0.05) the effects of spiramycin on the growth and antioxidant responses of Microcystis aeruginosa as well as the biodegradation of spiramycin by M. aeruginosa during a 7-day exposure test. At a nitrogen level of 0.5 mg L⁻¹, the activities of superoxide dismutase and catalase were stimulated by 100–400 ng L⁻¹of spiramycin to protect algal cells from oxidative damage, resulting in alleviated toxicity of spiramycin and low malondialdehyde content in M. aeruginosa. The catalase activity was inhibited by 400 ng L⁻¹of spiramycin at higher nitrogen levels of 5–50 mg L⁻¹, leading to significant growth inhibition (p < 0.05) and higher malondialdehyde content through accumulation of hydrogen peroxide. Stimulated glutathione content and glutathione S-transferase activity were coupled to the biodegradation of spiramycin in M. aeruginosa. The 7-day biodegradation percentage of spiramycin varied from 8.9 to 29.6 %, which was enhanced by increased nitrogen concentration and decreased spiramycin concentration. Due to the regulation of algal growth, the toxicity of M. aeruginosa were significantly enhanced (p < 0.05) by 100 ng L⁻¹of spiramycin at a nitrogen concentration of 0.5 mg L⁻¹while significantly reduced (p < 0.05) by 400 ng L⁻¹of spiramycin at nitrogen levels of 5–50 mg L⁻¹, according to the luminescent bacteria test. Low concentration of coexisting spiramycin contaminant should be considered during the control of M. aeruginosa bloom, especially under nitrogen deficient condition.

Efficacy of activated bentonite, activated carbon, and natural phosphate under experimental conditions was tested as low-cost adsorbents for spiramycin antibiotic removal from aqueous solution. Equilibrium kinetic and isotherm adsorption process are well described by pseudo-second order and Langmuir isotherm models for activated bentonite and activated carbon, while natural phosphate follows pseudo-first order and Freundlich models, respectively. Obtained results revealed that activated bentonite has the highest adsorption capacity (260.3 mg/g) as compared to activated carbon (80.3 mg/g) and natural phosphate (1.7 mg/g). The adsorption capacity decreases for all adsorbents in the presence of NaCl. The adsorption processes are facilitated in the alkaline pH range for activated bentonite and activated carbon, whereas, for natural phosphate, the acidic pH range is favorable. They are involving ion exchange and hydrogen bond mechanisms as well as Van der Waals forces and also π interactions for activated carbon. Thermodynamic calculation shows that spiramycin adsorption is endothermic and spontaneous on all adsorbents. The activated bentonite reusability is more efficient by more than 95% in two-step desorption using NaOH and HCl eluents compared to activated carbon. Thus, activated bentonite is a promising adsorbent for spiramycin removal from aqueous solution.

The main goal of this preliminary study was to quantify airborne particles and characterize the dominant cultivable bacterial species as well as some Gram-positive species, and their antibiotic resistance pattern, from environmental samples taken inside and outside of a dairy milking parlour. Sampling was performed over 2 days, in different seasons. The small viable particulate matter < 10 μm (bioaerosols) and cultivable bacteria reached their highest concentrations in the milking parlour. The majority of airborne bacteria in the milking parlour belonged to the genera Staphylococcus (41.9%) and Bacillus (20.9%). A total of 32 different bacterial species of Staphylococcus, Aerococcus, Bacillus, Pseudomonas, Serratia and Acinetobacter were identified. Many of these bacteria may be opportunistic pathogens, causing disease in humans or animals. We found low levels of acquired resistance to the antibiotics commonly used in human or animal infections caused by these opportunistic bacteria. More specifically, resistance to tetracyclines (13.4%), penicillin G (13.4%) and macrolides (7.5%) was identified in Staphylococcus sp. as was a methicillin-resistant S. hominis and resistance to spiramycin (n = 1), lincomycin (n = 1) and streptomycin (n = 2) in Aerococcus sp. An assessment of the occupational risk run by dairy farmers for contracting infections after long- or short-term exposure to micro-organisms requires further studies on the concentration of opportunistic pathogenic bacteria in dairy farm environments.

The interactive effects of binary antibiotic mixtures of spiramycin (SP) and ampicillin (AMP) on Microcystis aeruginosa (MA) in terms of growth as well as microcystin production and extracellular release were investigated through the response surface methodology (RSM). SP with higher 50 and 5% effective concentrations in MA growth was more toxic to MA than AMP. RSM model for toxic unit approach suggested that the combined toxicity of SP and AMP varied from synergism to antagonism with SP/AMP mixture ratio decreasing from reversed equitoxic ratio (5:1) to equitoxic ratio (1:5). Deviations from the prediction of concentration addition (CA) and independent action (IA) model further indicated that combined toxicity of target antibiotics mixed in equivalent ratio (1:1) varied from synergism to antagonism with increasing total dose of SP and AMP. With the increase of SP/AMP mixture ratio, combined effect of mixed antibiotics on MA growth changed from stimulation to inhibition due to the variation of the combined toxicity and the increasing proportion of higher toxic component (SP) in the mixture. The mixture of target antibiotics at their environmentally relevant concentrations with increased total dose and SP/AMP mixture ratio stimulated intracellular microcystin synthesis and facilitated MA cell lysis, thus leading to the increase of microcystin productivity and extracellular release.