Monobutyl phthalate (MBP) is the main metabolite of dibutyl phthalate (DBP) in vivo. MBP has a stable structure, can continuously accumulate in living organisms, and has the potentially to harm animal and human reproductive function. In the ovarian follicle microenvironment, MBP may lead to defects in follicular development and steroid production, abnormal meiotic maturation, impaired ovarian function and other reproductive deficits. In this study, SMART-seq was used to investigate the effects of MBP exposure on the in vitro maturation (IVM) and development of porcine oocytes. The results showed that differentially expressed genes after MBP exposure were enriched in the biological processes cytoskeleton, cell apoptosis, endoplasmic reticulum (ER) and mitochondria. Glycine (Gly) improved the developmental potential of porcine oocytes by regulating mitochondrial and ER function. The effect of Gly in protecting oocytes against MBP-induced damage was studied. The results showed that the addition of Gly significantly decreased the rate of MBP-induced spindle abnormalities, decreased the frequency of MBP-induced mitochondria-associated ER membrane (MAM) interactions, and downregulated the protein and gene expression of the linkage molecules Mitofusin 1 (MFN1) and Mitofusin 2 (MFN2) in the MAM. Additionally, treatment with Gly restored the distribution of the 1,4,5-triphosphate receptor 1 (IP₃R1) and voltage-dependent anion channel 1 (VDAC1), further decreasing the intracellular free calcium concentration ([Ca²⁺]ᵢ) levels and mitochondrial Ca²⁺ ([Ca²⁺]ₘ) , increasing the ER Ca²⁺ ([Ca²⁺]ER) levels, and thus significantly increasing the ER levels and mitochondrial membrane potential (ΔΨ m). Gly also decreased the levels of reactive oxygen species (ROS) and increased the levels of Glutathione (GSH), oocyte apoptosis-related indicators (Caspase-3 activity and Annexin V) and oocyte apoptosis-related genes (BAX, Caspase 3 and AIFM1). Our results suggest that Gly can ameliorate microtubule cytoskeleton abnormalities and improve oocyte maturation by reducing the defective mitochondrial–ER interactions caused by MBP exposure in vitro.

Antibiotics usage in animal production is considered a primary driver of the occurrence, supply and spread of antibiotic resistance genes (ARGs) in the environment. Pig farms and fish ponds are important breeding systems in food animal production. In this study, we compared and analyzed broad ARGs profiles, mobile genetic elements (MGEs) and bacterial communities in a representative pig farm and neighboring fish ponds around Poyang Lake, the largest freshwater lake in China. The factors influencing the distribution of ARGs were also explored. The results showed widespread detection of ARGs (from 57 to 110) among 283 targeted ARGs in the collected water samples. The differences in the number and relative abundance of ARGs observed from the pig farm and neighboring fish ponds revealed that ARG contamination was more serious on the pig farm than in the fish ponds and that the water treatment plant on the pig farm was not very effective. Based on the variance partition analysis (VPA), MGEs, bacterial communities and water quality indicators (WIs) codrive the relative abundance of ARGs. Based on network analysis, we found that total phosphorus and Tp614 were the most important WIs and MGEs affecting ARG abundance, respectively. Our findings provide fundamental data on farms in lakeside districts and provide insights into establishing standards for the discharge of aquaculture wastewater.

Antibiotic resistance genes (ARGs) are abundantly shed in feces. Thus, it is crucial to identify their host sources so that ARG pollution can be effectively mitigated and aquatic ecosystems can be properly conserved. Here, spatiotemporal variations and sources of ARGs in the Longjiang watershed of South China were investigated by linking them with microbial source tracker (MST) indicators. The most frequently detected ARGs (>90%) were sulI, sulII, blaTEM, tetW, ermF, and the mobile element intI1. Spatial distribution analyses showed that tributaries contributed significantly more sulI, sulII, and ermF contamination to the Longjiang watershed than the main channel. MST indicator analysis revealed that the Longjiang watershed was contaminated mainly by human fecal pollution. Livestock- and poultry-associated fecal pollution significantly declined after the swine fever outbreak. The occurrence of most ARGs is largely explained by human fecal pollution. In contrast, pig fecal pollution might account for the prevalence of tetO. Moreover, combined human-pig fecal pollution contributed to the observed blaNDM₋₁ distribution in the Longjiang watershed. Subsequent analysis of the characteristics of MST markers disclosed that the relatively lower specificities of BacHum and Rum-2-Bac may lead to inaccurate results of tracking ARG pollution source. The present study determined spatiotemporal variations and ARG origins in the Longjiang watershed by combining MST markers. It also underscored the necessity of using multiple MST markers simultaneously to identify and characterize ARG pollution sources accurately.

We determined the distribution and temporal variation of Carbapenem Resistant Enterobacterales (CRE), carbapenemase-encoding genes and other antibiotic resistance genes (ARGs) in a highly polluted river (Lis River; Portugal), also assessing the potential influence of water quality to this distribution. Water samples were collected in two sampling campaigns performed one year apart (2018/2019) from fifteen sites and water quality was analyzed. CRE were isolated and characterized. The abundance of four ARGs (blaNDM, blaKPC, tetA, blaCTX₋M), two Microbial Source Tracking (MST) indicators (HF183 and Pig-2-Bac) and the class 1 integrase gene (IntI1) was measured by qPCR. confirmed the poor quality of the Lis River water, particularly in sites near pig farms. A collection of 23 CRE was obtained: Klebsiella (n = 19), Enterobacter (n = 2) and Raoultella (n = 2). PFGE analysis revealed a clonal relationship between isolates obtained in different sampling years and sites. All CRE isolates exhibited multidrug resistance profiles. Klebsiella and Raoultella isolates carried blaKPC while Enterobacter harbored blaNDM. Conjugation experiments were successful for only four Klebsiella isolates. All ARGs were detected by qPCR on both sampling campaigns. An increase in ARGs and IntI1 abundances was detected in sites located downstream of wastewater treatment plants. Strong correlations were observed between blaCTX₋M, IntI1 and the human-pollution marker HF183, and also between tetA and the pig-pollution marker Pig-2-bac, suggesting that both human- and animal-derived pollution in the Lis River are a potential source of ARGs. Plus, water quality parameters related to eutrophication and land use were significantly correlated with ARGs abundances. Our findings demonstrated that the Lis River encloses high levels of antibiotic resistant bacteria and ARGs, including CRE and carbapenemase-encoding genes. Overall, this study provides a better understanding on the impacts of water pollution resulting from human and animal activities on the resistome of natural aquatic systems.

Veterinary pharmaceuticals, including antibiotics, are emerging contaminants of concern worldwide. Avian scavengers are exposed to pharmaceuticals through consumption of livestock carcasses used for feeding wildlife for conservation purposes at supplementary feeding stations. Here we tested the hypothesis that griffon vultures (Gyps fulvus) would be more exposed to antibiotics (i.e., quinolones) when feeding on livestock carcasses from intensive farming than when they rely on carcasses from extensive farming or wild animals. We sampled 657 adult griffon vultures captured between 2008 and 2012. In addition, we sampled tissues from domestic livestock supplied at feeding stations in the study area between 2009 and 2019; pig (n = 114), sheep (n = 28), cow (n = 1) and goat (n = 2). Samples were analysed by liquid chromatography with electrospray ionization mass spectrometry (LC-ESI-MS). Quinolones were detected in plasma from 12.9% of the griffon vultures analysed. Quinolone prevalence in griffon vultures varied significantly among feeding stations but was also affected by the total amount of carcasses supplemented, especially the mass of pig carcasses. These results aligned with a 21.1% quinolone prevalence in pig carcasses sampled at feeding stations, wherein enrofloxacin and ciprofloxacin levels of up to 3359 ng/g and 1550 ng/g, respectively, were found. Given enrofloxacin pharmacokinetics in pig tissues, 5.3% of the analysed pigs may have died no more than one day after treatment. Quinolone presence in vultures was negatively associated with blood lead levels, which mostly originates from lead ammunition and indicates a higher consumption of game animal carcasses. Carcass disposal for feeding avian scavengers must always assess and manage the risks posed by veterinary pharmaceuticals, especially when livestock provided may have died soon after treatment.

A high demand exists in bisphenols (BPs) screening studies for quick, reliable and straightforward analytical methods that generate data faster and simultaneously. Herein, we describe a combination of enzymatic probe sonication (EPS) and ultra-performance liquid chromatography-tandem mass spectrometry (UPLC-MS/MS) for quick extraction and simultaneous quantification of eight important BPs in animal-derived foods. Results obtained demonstrated that the ultrasonic probe power could not only enhance the enzymatic hydrolysis efficiency, but also accelerate the liquid-liquid extraction procedure. Under optimized EPS parameters, one sample could be exhaustively extracted within 120 s, as compared with 12 h needed for the conventional enzymatic extraction which is more suitable for high-throughput analysis. The method was successfully applied to analyze residual BPs in animal-derived foods collected from Beijing, China. Widespread occurrence of BPA, BPS, BPF, BPAF, BPP, and BPB were found, with detection frequencies of 65.2%, 42.4%, 33.7%, 29.4%, 28.3%, and 27.2%, respectively. The highest total concentration levels of BPs (sum of the eight BPs analyzed, ΣBPs) were found in chicken liver (mean 12.2 μg/kg), followed by swine liver (6.37 μg/kg), bovine muscle (3.24 μg/kg), egg (2.03 μg/kg), sheep muscle (2.03 μg/kg), chicken muscle (1.45 μg/kg), swine muscle (1.42 μg/kg), and milk (1.17 μg/kg). The estimated daily intake (EDI) of BPs, based on the mean and 95th percentile concentrations and daily food consumptions, was estimated to be 5.687 ng/kg bw/d and 22.71 ng/kg bw/d, respectively. The human health risk assessment in this work suggests that currently BPs do not pose significant risks to the consumers because the hazard index (HI) was <1.

The rapid expansion of organic rice cultivation areas have been accompanied by increased application of organic fertilizers. The high prevalence of soil antibiotic resistance caused by organic fertilizer application poses a severe threat to the agricultural and soil ecosystems. To date, research efforts and understanding of the effects and mechanism of action of the various organic fertilizers on antibiotic resistance in paddy soils remain poorly investigated. Tetracycline resistance genes (TRGs, including tetB, tetC, tetL, tetZ, tetM, tetO, tetT, and tetX), class 1 integron-integrase gene (intI1) and bacterial communities were characterized using quantitative-PCR and Illumina MiSeq sequencing, in paddy soils exposed to inorganic fertilizer (NPK), animal-derived organic fertilizer (AOF, composted swine and/or chicken manure), plant-derived organic fertilizer (POF, rapeseed cake and/or astragalus) and commercial organic fertilizer (COF, composted of animal manure mix with crop residues) applications. Compared with NPK, AOF applications significantly increased the relative abundance of TRGs, which was predominantly expressed in the increase of the relative abundance of tetC, tetM, tetO, tetT, and tetX, while POF and COF had no significant effect on the relative abundance of TRGs. Principal coordinate analysis revealed that AOF and POF significantly altered bacterial communities in paddy soils relative to NPK, while COF had no significant change of bacterial communities. Variation partitioning analysis indicated that soil physicochemical properties were the decisive factors for the changes of TRGs in organic paddy fields. Furthermore, redundancy analysis and the Mantel test showed that TRG profiles in AOF applied paddy soils were strongly influenced by electrical conductivity (EC). Total nitrogen (TN) and organic matter (OM) affected the distribution of TRGs in COF and POF applied paddy soils through a different mechanism. This study provides insights into the impacts of different types of organic fertilizer on the profiles of TRGs in paddy soils.

Biological treatment is one of the most widely used methods to treat swine wastewater in wastewater treatment plants. The microbial community plays an important role in the swine slurry treatment system. However, limited information is available regarding the correlation between pollutant concentration and dominant microbial community in swine wastewater. This work aimed to study the profiling of microbial communities and their abundance in the 40 M³/day large-scale and step-by-step treatment pools of swine wastewater. Metagenome sequencing was applied to study the changes of microbial community structure in biochemical reaction pools. The results showed that in the heavily polluted pools, it was mainly Proteobacteria, Cyanobacteria, Chlorella and other strains that could tolerate high concentration of ammonia nitrogen to remove nitrogen and absorb chemical oxygen demand (COD). In the moderately polluted pools, Nitrospirae, Actinobacteria and other strains further cooperated to purify swine wastewater. In the later stage, the emergence of Brachionus indicated the reduction of water pollution. The dominant microbes and their abundance changed with the purification of swine wastewater in different stages. Moreover, the dominant microflora of swine wastewater treatment pools at all levels reflected little difference in phylum classification level, while in genus classification level, the dominant microflora manifested great difference. Findings demonstrated that the microorganisms maintained ecological balance and absorbed the nutrients in the swine wastewater treatment pools, so as to play the role of purifying sewage. Therefore, the stepwise purification of swine wastewater can be realized by adding bacteria and microalgae of different genera.

In many countries, the adverse impact of agriculture on water sources has been discussed with more attention recently by the water footprint estimation. Brazil is the second largest animal protein’ exporter, and this demand has a tendency to increase significantly until 2050, and in this context the water management will be crucial. Thus, the present study aimed to evaluate the water footprint and productivity in the broiler and swine slaughtered in Brazil from 2008 to 2018. The results showed that the herds of broiler and swine were concentrated in three main regions: Midwest, Southeast and South, representing 97.1% of the broilers and 99.7% of the swine slaughtered in Brazil. During the studied decade, the slaughter of broiler and swine increased 9.1 and 25.8%, respectively. The broiler and swine water footprint decreased by 15.4 and 3.5%, respectively. The average volume of water needed for the production of broiler and swine meat was 2533 L kg⁻¹ and 3754 L kg⁻¹, respectively. The average water productivity per kg of broiler meat was 0.397 kg m⁻³, while for swine it was 0.269 kg m⁻³. The average water productivity for soybean was 0.497 kg m⁻³, and for corn it was 1.18 kg m⁻³. The decrease in the water footprint is a result of the improvement of management practices, highlighting that it is necessary to improve the knowledge about the use of the water footprint methodology as a tool for water management to help public policies.

Swine wastewater (SW) poses a great threat to the environment due to its high-nutrient profiles if not properly managed. Advanced biological treatment method is an efficient method to treat SW by screening potent microalgae or bacterial strains. In this study, activated sludge, three locally isolated heterotrophic nitrification bacteria and one microalgal strain (Chlorella) were used as inoculums in treating a local SW. Their treatment efficiencies were compared, while the nitrogen removal mechanisms and microbiome profile were explored in detail. It was found that certain heterotrophic nitrification strains had a slight advantage in removing chemical oxygen demand and phosphorus from SW, with the highest removal efficiencies of 83.9% and 76.2%, respectively. The removal efficiencies of ammonia nitrogen and total nitrogen in wastewater by microalgae reached 80.9% and 66.0% respectively, which were far higher than all the heterotrophic nitrification strains. Biological assimilation was the main pathway of nitrogen conversion by microalgae and heterotrophic nitrifying bacteria; especially microalgae showed excellent biological assimilation performance. Correlation analysis showed that Comamonas was highly positively correlated with nitrogen assimilation, while Acidovorax was closely correlated with simultaneous nitrification and denitrification. This study gives a comparison of microalgae and heterotrophic nitrifying bacteria on the nitrogen transfer and transformation pathways.