Tetracyclines, sulphonamides, and quinolones are families of antimicrobials (AMs) widely used in the poultry industry and can excrete up to 90% of AMs administrated, which accumulate in poultry litter. Worryingly, poultry litter is widely used as an agriculture fertilizer, contributing to the spread AMs residues in the environment. The aim of this research was to develop a method that could simultaneously identify and quantify three AMs families in poultry litter by high-performance liquid chromatography–tandem mass spectrometry (HPLC–MS/MS). Samples of AMs free poultry litter were used to validate the method according to 657/2002/EC and VICH GL49. Results indicate that limit of detection (LOD) ranged from 8.95 to 20.86 μg kg⁻¹, while limits of quantitation (LOQ) values were between 26.85 and 62.58 µg kg⁻¹ of tetracycline, 4-epi-tetracycline, oxytetracycline, 4-epi-oxytetracycline, enrofloxacin, ciprofloxacin, flumequine, sulfachloropyridazine, and sulfadiazine. Recoveries obtained ranged from 93 to 108%. The analysis of field samples obtained from seven commercial poultry flocks confirmed the adequacy of the method since it detected means concentrations ranging from 20 to 10,364 μg kg⁻¹. This provides us an accurate and reliable tool to monitor AMs residues in poultry litter and control its use as agricultural fertilizer.

The monk parakeet (Myiopsitta monachus) is considered to be one of the most invasive bird species because its unique ability among parrots to build their own communal nests. Currently, they are considered an invasive species in 19 countries and a pest—even in their native distribution—because of economic losses derived from their impacts. During the reproductive seasons of 2017 and 2018, we registered interactions between invasive monk parakeets and resident bird species in Santiago, Chile. We observed agonistic and affiliative interactions, and further, we described monk parakeets’ nest occupancy by nine bird species, two invasive and seven native. For this reason, we consider that the monk parakeet is an allogenic ecosystem engineer with the potential to shape distribution and richness of sympatric species in urban environments. Our results contribute to an assessment of the implications of the monk parakeet’s ecological invasion to other synanthropic species, and raise concern of other potential impacts, such as pathogen transmission derived from these interactions.

Tetracyclines are important antimicrobial drugs for poultry farming that are actively excreted via feces and urine. Droppings are one of the main components in broiler bedding, which is commonly used as an organic fertilizer. Therefore, bedding becomes an unintended carrier of antimicrobial residues into the environment and may pose a highly significant threat to public health. For this depletion study, 60 broiler chickens were treated with 20% chlortetracycline (CTC) under therapeutic conditions. Concentrations of CTC and 4-epi-CTC were then determined in their droppings. Additionally, this work also aimed to detect the antimicrobial activity of these droppings and the phenotypic susceptibility to tetracycline in E. coli isolates, as well as the presence of tet(A), tet(B), and tet(G) resistance genes. CTC and 4-epi-CTC concentrations that were found ranged from 179.5 to 665.8 µg/kg. Based on these data, the depletion time for chicken droppings was calculated and set at 69 days. All samples presented antimicrobial activity, and a resistance to tetracyclines was found in bacterial strains that were isolated from these samples. Resistance genes tet(A) and tet(B) were also found in these samples.

Oxytetracycline (OTC), enrofloxacin (EFX), and sulfachloropyridazine (SCP) are critically important antimicrobials (AMs) in both human and veterinary medicine, where they are widely used in farm animals. Lettuce has become a matrix of choice for studying the presence of residues of these AMs in plants, as the concentrations of residues detected in lettuce can range from ng to mg. While several analytical methodologies have been developed for the purpose of detecting AMs in lettuce, these currently do not detect both the parent compound and its active metabolites or epimers, such as in the case of ciprofloxacin (CFX) and 4-epi-oxitetracycline (4-epi-OTC), which also pose a risk to public health and the environment due to their AM activity. In light of this situation, this work proposes an analytical method that was developed specifically to allow for the detection of OTC, 4-epi-OTC, EFX, CFX, and SCP in a lettuce matrix. This method uses acetonitrile, methanol, 0.5% formic acid, and McIlvaine-EDTA buffer as extraction solvents, and dispersive solid-phase extraction (dSPE) for the clean-up. The analytes were detected using a liquid chromatography technique coupled to mass spectrometry (HPLC-MS/MS). Parameters such as the specificity, linearity, recovery, precision, limit of detection, and limit (LOD) of quantification (LOQ) were calculated according to the recommendations established in the European Union decision 2021/808/EC and VICH GL2: Validation of analytical procedures. The LOQ for the analytes OTC, 4-epi-OTC, CFX, and SCP was 1 μg·kg⁻¹, whereas for EFX, it was 5 μg·kg⁻¹ dry weight. All calibration curves showed a coefficient of determination (R²) of >0.99. The recovery levels ranged from 93.0 to 110.5% and the precision met the acceptance criteria, with a coefficient of variation of ≤14.02%. Therefore, this methodology allows for the precise and reliable detection and quantification of these analytes. The analysis of commercial samples confirmed the suitability of this method.

Oxytetracycline (OTC) is widely used in broiler chickens. During and after treatment a fraction of OTC is excreted in its original form and as its epimer, 4-epi-OTC in droppings. To address the transfer of OTC into the environment, we evaluated the dissemination of OTC and 4-epi-OTC from treated birds to the environment and sentinels, through the simultaneous analysis of broiler droppings and litter. Male broiler chickens were bred in controlled conditions. One group was treated by orogastric tube with 80 mg kg⁻¹ of OTC and two groups received no treatment (sentinels). OTC+4-epi-OTC were analyzed and detected by a HPLC-MS/MS post the end of treatment. The highest concentrations of OTC+4-epi-OTC were detected in the droppings of treated birds 14-days following the end of treatment (2244.66 µg kg⁻¹), and one day following the end of treatment in the litter (22,741.68 µg kg⁻¹). Traces of OTC+4-epi-OTC were detected in the sentinels’ droppings and litter (<12.2 µg kg⁻¹). OTC+4-epi-OTC can be transferred from treated birds to the environment and to other untreated birds. The presence and persistence of OTC+4-epi-OTC in litter could contribute to the selection of resistant bacteria in the environment, increasing the potential hazard to public and animal health.

Monk Parakeets (Myiopsitta monachus) are medium-sized parrots that due to international pet trade currently exist as invasive species in 19 countries globally. Such is the case of Chile, where Monk Parakeets have thrived in the city of Santiago. Cryptosporidium spp. and Giardia spp. are worldwide distributed gastrointestinal parasites whose potential hosts include birds and humans. The present study sought to determine the presence of these pathogens in Monk Parakeets from Santiago. During the austral summers of 2017 and 2018, 207 Monk Parakeet nestlings were captured, and fecal samples were studied via microscopical analyses. Environmental data related to the trees in which the nestlings were captured were analyzed to establish the existence of infection clusters. Associations between spatial clusters, environmental variables, and the presence or absence of these pathogens were explored. In total, 33 samples were positive to the presence of one or both protozoa. Of the 33, Cryptosporidium spp. oocysts were detected in 10 nestlings (30%) while Giardia spp. cysts were detected in 25 (76%). Two nestlings presented poly-parasitism (6%). Statistical analyses established pruned trees as a potential protective factor against infection with these parasites. The present study corresponds to the second report of Cryptosporidium spp. in Monk Parakeets in Chile and the first worldwide report of Giardia spp. in these birds, emphasizing Monk Parakeet’s potential role as a reservoir and pathogen disseminator, especially in urban environments.