Manure Samples Research Articles

Impact of soil supplemented with pig manure on the abundance of antibiotic resistant bacteria and their associated genes.

This study was conducted to evaluate the abundance of antibiotic resistant bacteria and their resistance genes from agriculture soil supplemented with pig manure. Uncultivable soil sample was supplemented with pig manure samples under microcosm experimental conditions and plated on Luria-Bertani (LB) agar incorporated with commercial antibiotics. The supplementation of soil with 15% pig manure resulted in the highest increase in the population of antibiotic resistant bacteria (ARB)/multiple antibiotic resistant bacteria (MARB). Seven genera that included Pseudomonas, Escherichia, Providencia, Salmonella, Bacillus, Alcaligenes and Paenalcaligenes were the cultivable ARB identified. A total of ten antibiotic resistant bacteria genes (ARGs) frequently used in clinical or veterinary settings and two mobile genetic elements (MGEs) (Class 1 and Class 2 integrons) were detected. Eight heavy metal, copper, cadmium, chromium, manganese, lead, zinc, iron, and cobalt were found in all of the manure samples at different concentrations. Tetracycline resistance genes were widely distributed with prevalence of 50%, while aminoglycoside and quinolone-resistance gene had 16% and 13%, respectively. Eighteen ARB isolates carried more than two ARGs in their genome. Class 1 integron was detected among all the 18 ARB with prevalence of 90-100%, while Class 2 integron was detected among 11 ARB. The two classes of integron were found among 10 ARB. Undoubtedly, pig manure collected from farms in Akure metropolis are rich in ARB and their abundance might play a vital role in the dissemination of resistance genes among clinically-relevant pathogens.

Effects of diets with different fibre content on the performance of rabbit does and on parasitological infection

The effect of two diets with different fibre contents was examined on rabbit does’ performance and parasitological infection. Diet P2 had 1% higher crude fibre, ADF and starch contents than that of diet P1. The performance of 12–12 does and litters per group were examined during two consecutive inseminations. In both dietary groups, according to a 49-day reproduction rhythm, the does were artificially inseminated 18 days after kindling. Most production traits were not affected by the diets (kindling rate, weight of does and their kits, feed intake, mortality; P>0.05). Significant differences were only found in litter sizes as the number of kits was lower in P1 diet fed group at all examined days of lactation. The significant “Diet x Reproduction cycle” interaction showed that the litter sizes of dietary groups did not differ at the first examined insemination, however at the subsequent reproduction cycle 6–8% lower litter sizes were counted in P1 fed does compared to group P2 (litter size at 4 day: 9.00 vs 9.58–9.92, P = 0.004; litter size at 11 day: 8.83 vs 9.58–9.92, P = 0.037 for diet P1 at the 2nd reproductive cycle compared to all the other cases, respectively). The two diets with different compositions did not affect most of the production results of the rabbit does and their kits, but the lower litter size of does consuming P1 diet at the second examined lactation suggests the adverse effect of P1 diet’s long-term use. From the collected manure samples not any parasites were detected which presents a very favourable picture of farm from the point of view of hygiene and animal health.

Assessment of the biochemical methane potential of in-house and outdoor stored pig and dairy cow manure by evaluating chemical composition and storage conditions

Biogas production is a suitable option for producing energy from dairy and pig manure types. During manure storage, organic matter degradation results in methane emissions decreasing the potential biogas yield. The present research advances the understanding of the biochemical methane potential (BMP) and the chemical characteristics of manure collected year-round from sequential stages of the liquid manure management chain of commercial dairy cow and pig farms. To this end, manure samples from six livestock farms in Germany were analyzed. The results showed that changes in chemical composition during storage led to a 20.5% decrease in the BMP of dairy manure from the barn to outdoor storage. For fattening pig manure samples, there was a 39.5% decrease in the BMP from intermediate to outdoor storage. An analysis of BMP according to manure age showed that pig manure degrades faster than dairy manure; the importance of promptly feeding manure to the biogas plant in order to avoid significant CH4 emission losses and reduction in energy producing capacity was highlighted. The best BMP predictors for dairy manure were the contents of dry matter, volatile solids and lignin, whereas best BMP predictors for pig manure were dry matter and volatile fatty acid (VFA) content. Prediction models performed well for samples from outdoor storages; refinements for predicting BMP of less aged samples presenting lower chemical variability would be necessary.

129 Case Study: Passage Dynamics of Whole Cottonseed in Beef Cattle

Abstract Whole cottonseed (WCS) has long been a viable supplement in southern beef production systems, but new cultivars and ginning innovations have renewed interest in its use. Ad libitum supplementation of WCS, however, leads to increased fat intake, potentially resulting in reduced fiber digestibility. However, gestating and lactating beef cows have been observed to have adequate or optimum performance under supplementation scenarios as much as 3.6 kg∙animal-1∙day-1. It has been hypothesized that WCS may pass through the gastrointestinal tract without being digested, thereby not interfering with whole diet digestibility. Thus, our objective was to conduct an observational screening of beef cattle offered near ad libitum levels of WCS for passage dynamics. In this case study, we sampled beef cattle (approximately 1,000 head) at a farm in central Alabama offered an average of 3.6 kg WCS∙animal-1∙day-1. Cattle were divided by age and stage of production across seven pastures varying in forage base. Fresh manure samples (n = 22) were collected at random from the herd. Fecal samples were sorted by wet sieving to recover undigested seed material. Counts were conducted to determine whole or fractional seed remaining. Data were analyzed using SAS v. 9.4. Across samples, recovered WCS were approximately normally distributed (W = 0.83) with a mean of 3.3 seed per sample and a range of 2 to 76. Fractional WCS recovered were normally distributed (W = 0.92) with a mean of 31.9 parts per sample and a range of 0 to4. On average, WCS recovered accounted for 0.34 g per sample, or 5.0% of the DM recovered. While caution should be used in broad interpretation of these observational results, there is sufficient evidence to suggest that high levels of WCS supplementation may seed to decreased digestive efficiency (evidenced by undigested seed in manure). These observations serve as a basis for further investigation in designed experiments to evaluate WCS digestion and passage dynamics.

Nitrogen transformation processes catalyzed by manure microbiomes in earthen pit and concrete storages on commercial dairy farms

Storing manure is an essential aspect of nutrient management on dairy farms. It presents the opportunity to use manure efficiently as a fertilizer in crop and pasture production. Typically, the manure storages are constructed as earthen, concrete, or steel-based structures. However, storing manure can potentially emit aerial pollutants to the atmosphere, including nitrogen and greenhouse gases, through microbial and physicochemical processes. We have characterized the composition of the microbiome in two manure storage structures, a clay-lined earthen pit and an aboveground concrete storage tank, on commercial dairy farms, to discern the nitrogen transformation processes, and thereby, inform the development of mitigation practices to preserve the value of manure. First, we analyzed the 16S rRNA-V4 amplicons generated from manure samples collected from several locations and depths (0.3, 1.2, and 2.1–2.75 m below the surface) of the storages, identifying a set of Amplicon Sequence Variant (ASVs) and quantifying their abundances. Then, we inferred the respective metabolic capabilities. These results showed that the manure microbiome composition was more complex and exhibited more location-to-location variation in the earthen pit than in the concrete tank. Further, the inlet and a location with hard surface crust in the earthen pit had unique consortia. The microbiomes in both storages had the potential to generate ammonia but lacked the organisms for oxidizing it to gaseous compounds. However, the microbial conversion of nitrate to gaseous N2, NO, and N2O via denitrification and to stable ammonia via dissimilatory nitrite reduction seemed possible; minor quantities of nitrate was present in manure, potentially originating from oxidative processes occurring on the barn floor. The nitrate-transformation linked ASVs were more prevalent at the near-surface locations and all depths of the inlet. Anammox bacteria and archaeal or bacterial autotrophic nitrifiers were not detected in either storage. Hydrogenotrophic Methanocorpusculum species were the primary methanogens or methane producers, exhibiting higher abundance in the earthen pit. These findings suggested that microbial activities were not the main drivers for nitrogen loss from manure storage, and commonly reported losses are associated with the physicochemical processes. Finally, the microbiomes of stored manure had the potential to emit greenhouse gases such as NO, N2O, and methane.

A longitudinal study to examine the influence of farming practices and environmental factors on pathogen prevalence using structural equation modeling.

The contamination of fresh produce with foodborne pathogens has been an on-going concern with outbreaks linked to these commodities. Evaluation of farm practices, such as use of manure, irrigation water source, and other factors that could influence pathogen prevalence in the farming environment could lead to improved mitigation strategies to reduce the potential for contamination events. Soil, water, manure, and compost were sampled from farms in Ohio and Georgia to identify the prevalence of Salmonella, Listeria monocytogenes (Lm), Campylobacter, and Shiga-toxin-producing Escherichia coli (STEC), as well as Arcobacter, an emerging human pathogen. This study investigated agricultural practices to determine which influenced pathogen prevalence, i.e., the percent positive samples. These efforts identified a low prevalence of Salmonella, STEC, and Campylobacter in soil and water (< 10%), preventing statistical modeling of these pathogens. However, Lm and Arcobacter were found in soil (13 and 7%, respectively), manure (49 and 32%, respectively), and water samples (18 and 39%, respectively) at a comparatively higher prevalence, suggesting different dynamics are involved in their survival in the farm environment. Lm and Arcobacter prevalence data, soil chemical characteristics, as well as farm practices and weather, were analyzed using structural equation modeling to identify which factors play a role, directly or indirectly, on the prevalence of these pathogens. These analyses identified an association between pathogen prevalence and weather, as well as biological soil amendments of animal origin. Increasing air temperature increased Arcobacter and decreased Lm. Lm prevalence was found to be inversely correlated with the use of surface water for irrigation, despite a high Lm prevalence in surface water suggesting other factors may play a role. Furthermore, Lm prevalence increased when the microbiome's Simpson's Diversity Index decreased, which occurred as soil fertility increased, leading to an indirect positive effect for soil fertility on Lm prevalence. These results suggest that pathogen, environment, and farm management practices, in addition to produce commodities, all need to be considered when developing mitigation strategies. The prevalence of Arcobacter and Lm versus the other pathogens suggests that multiple mitigation strategies may need to be employed to control these pathogens.

Determination of Essential Minerals in Manures of Selected Domestic Animals in Karatina and Their Effects on Soil and Plants

Calcium and Magnesium are part of the macro-nutrients required by plants from the soil for growth and development, they are also necessary for soil health to keep crops healthy. Calcium is needed in plants for the formation of new cells, roots, stems, and leaves. For the soil, calcium plays an important role in ensuring the Pretention capacity of water and nutrients. Magnesium is an important component of photosynthesis in plants and is found in chlorophyll which,the green pigment. The study investigated and compared the presence of calcium and magnesium levels in manures of several animals from different households around the Kagochi area. The samples collected were dried, blended and acid digested. AAS, A-A-700,0 (Atomic Absorption Spectroscopy) was used to analyze the manure samples. The data provided was presented in tables and graphs comparing the manures from both households. The mean levels of calcium and magnesium for both households one and two were found to be Ca 27.99 mg/L and Mg 6.23mg/L for household 1 and Ca 27.26 mg/L and Mg 6.32 mg/L for household 2. A strong correlation was found between manures in household one and those of household two in their calcium and magnesium concentrations. The manures were found to contribute a significant rise in soil pH. The information from the study will form a basis for manure’s importance and application for small-scale and large-scale crop growers and bring more value to manures as a source of plant and soil nutrients. Keywords: Manure, Calcium, Magnesium, Nutrients, PH, Fertilizer. DOI: 10.7176/CMR/15-1-03 Publication date: May 31 st 2023

Risk factors associated with the prevalence of Shiga-toxin-producing Escherichia coli in manured soils on certified organic farms in four regions of the USA

IntroductionBiological soil amendments of animal origin (BSAAO), including untreated amendments are often used to improve soil fertility and are particularly important in organic agriculture. However, application of untreated manure on cropland can potentially introduce foodborne pathogens into the soil and onto produce. Certified organic farms follow the USDA National Organic Program (NOP) standards that stipulate a 90- or 120-day interval between application of untreated manure and crop harvest, depending on whether the edible portion of the crop directly contacts the soil. This time-interval metric is based on environmental factors and does not consider a multitude of factors that might affect the survival of the main pathogens of concern. The objective of this study was to assess predictors for the prevalence of Shiga-toxin-producing Escherichia coli (non-O157 STEC) in soils amended with untreated manure on USDA-NOP certified farms.MethodsA longitudinal, multi-regional study was conducted on 19 farms in four USA regions for two growing seasons (2017–2018). Untreated manure (cattle, horse, and poultry), soil, and irrigation water samples were collected and enrichment cultured for non-O157 STEC. Mixed effects logistic regression models were used to analyze the predictors of non-O157 STEC in the soil up to 180 days post-manure application.Results and discussionResults show that farm management practices (previous use with livestock, presence of animal feces on the field, season of manure application) and soil characteristics (presence of generic E. coli in the soil, soil moisture, sodium) increased the odds of STEC-positive soil samples. Manure application method and snowfall decreased the odds of detecting STEC in the soil. Time-variant predictors (year and sampling day) affected the presence of STEC. This study shows that a single metric, such as the time interval between application of untreated manure and crop harvest, may not be sufficient to reduce the food safety risks from untreated manure, and additional environmental and farm-management practices should also be considered. These findings are of particular importance because they provide multi-regional baseline data relating to current NOP wait-time standards. They can therefore contribute to the development of strategies to reduce pathogen persistence that may contribute to contamination of fresh produce typically eaten raw from NOP-certified farms using untreated manure.

Comparison of antibiotic resistance genes in swine manure storage pits of Iowa, USA

Antimicrobial resistance (AMR) can develop in deep-pit swine manure storage when bacteria are selectively pressured by unmetabolized antibiotics. Subsequent manure application on row crops is then a source of AMR into soil and downstream runoff water. Therefore, understanding the patterns of diverse antibiotic resistance genes (ARGs) in manure among different farms is important for both interpreting the results of the detection of these genes from previous studies and for the use of these genes as bioindicators of manure borne antibiotic resistance in the environment. Previous studies of manure-associated ARGs are based on limited samples of manures. To better understand the distribution of ARGs between manures, we characterized manures from 48 geographically independent swine farms across Iowa. The objectives of this study were to characterize the distribution of ARGs among these manures and to evaluate what factors in manure management may influence the presence of ARGs in manures. Our analysis included quantification of two commonly found ARGs in swine manure, ermB and tetM. Additionally, we characterized a broader suite of 31 ARGs which allowed for simultaneous assays of the presence or absence of multiple genes. We found the company integrator had a significant effect on both ermB (P=0.0007) and tetM gene concentrations (P=0.0425). Our broad analysis on ARG profiles found that the tet(36) gene was broadly present in swine manures, followed by the detection of tetT, tetM, erm(35), ermF, ermB, str, aadD, and intl3 in samples from 14 farms. Finally, we provide a comparison of methods to detect ARGs in manures, specifically comparing conventional and high-throughput qPCR and discuss their role in ARG environmental monitoring efforts. Results of this study provide insight into commonalities of ARG presence in manure holding pits and provide supporting evidence that company integrator decisions may impact ARG concentrations.

Draft Genome Sequences of 110 Shiga Toxin-Producing Escherichia coli Isolates Collected from Bovine Manure Samples in the Southern United States.

Shiga toxin-producing Escherichia coli (STEC) bacteria continue to impact the food industry. Environmental sampling of potential sources of contamination is important to aid epidemiologic efforts in tracking foodborne illnesses throughout the United States. Here, the draft genome sequences of 110 STEC isolates from bovine manure collected in Florida and Texas are reported.

Genotyping-driven diversity assessment of biocontrol potent Bacillus spp. strain collection as a potential method for the development of strain-specific biomarkers.

Bacillus species are among the most researched and frequently applied biocontrol agents. To estimate their potential as environmentally friendly microbial-based products, reliable and rapid plant colonization monitoring methods are essential. We evaluated repetitive element-based (rep) and Random Amplified Polymorphic DNA (RAPD) PCR (Polymerase Chain Reaction) genotyping in a diversity assessment of 251 strains from bulk soil, straw, and manure samples across Serbia, highlighting their discriminative force and the presence of unique bands. RAPD 272, OPG 5, and (GTG)5 primers were most potent in revealing the high diversity of a sizable environmental Bacillus spp. collection. RAPD 272 also amplified a unique band for a proven biocontrol strain, opening the possibility of Sequence Characterized Amplified Region (SCAR) marker design. That will enable colonization studies using the SCAR marker for its specific detection. This study provides a guide for primer selection for diversity and monitoring studies of environmental Bacillus spp. isolates.

Metagenomic analysis reveals patterns and hosts of antibiotic resistance in different pig farms.

In actual production environments, antibiotic-resistant genes (ARGs) are abundant in pig manure, which can form transmission chains through animals, the environment, and humans, thereby threatening human health. Therefore, based on metagenomic analysis methods, ARGs and mobile genetic elements (MGEs) were annotated in pig manure samples from 6 pig farms in 3 regions of Shanxi Province, and the potential hosts of ARGs were analyzed. The results showed that a total of 14 ARG types were detected, including 182 ARG subtypes, among which tetracycline, phenol, aminoglycoside, and macrolide resistance genes were the main ones. ARG profiles, MGE composition, and microbial communities were significantly different in different regions as well as between different pig farms. In addition, Anaerobutyricum, Butyrivibrio, and Turicibacter were significantly associated with multiple ARGs, and bacteria such as Prevotella, Bacteroides, and the family Oscillospiraceae carried multiple ARGs, suggesting that these bacteria are potential ARG hosts in pig manure. Procrustes analysis showed that bacterial communities and MGEs were significantly correlated with ARG profiles. Variation partitioning analysis results indicated that the combined effect of MGEs and bacterial communities accounted for 64.08% of resistance variation and played an important role in ARG profiles. These findings contribute to our understanding of the dissemination and persistence of ARGs in actual production settings, and offer some guidance for the prevention and control of ARGs contamination.

Evaluation of Environmental Sampling for Detection of Mycobacterium avium subspecies paratuberculosis in the Pre-Weaned Calf Area and Calving Area of Infected Dairy Farms Enrolled in a Voluntary Johne’s Disease Control Programme

One of the pillars of Johne's disease control is to break the cycle of transmission from infectious adult animals to young susceptible animals. Many control programmes utilise a risk assessment and management plan (RAMP) to identify specific risks for transmission of infection on individual farms and then recommend tailored biosecurity measures to mitigate the risk. It is important that infected farms in particular, practice effective biocontainment in the calving area and pre-weaned calf area. The objectives of this study were: (1) to determine what proportion of MAP-infected farms (PCR-positive) enrolled in a control programme had detectable MAP in their calf environment; (2) to compare RAMP scores and apparent within-herd prevalence (aWHP) of PCR-positive and PCR-negative farms; (3) to evaluate the correlation between RAMP scores, aWHP and levels of contamination based on PCR test Ct value. A novel environmental sampling protocol combining manure samples and boot swab samples was conducted in the calving area and pre-weaned calf area on 28 dairy farms with PCR-confirmed MAP infection. All samples were tested with PCR. Logistic regression modelling was used to evaluate the association between RAMP score and aWHP and PCR outcome. Overall, 46% of farms had positive PCR samples in either the calving area, pre-weaned calf area or both. The calving area was positive in 36% of farms and the pre-weaned calf area was positive in 21% of farms. There was a moderate, negative correlation (r = -0.45; p = 0.08) observed between RAMP scores and Ct value at the sample-level that slightly missed the required significance level. No significant association was identified between RAMP scores or aWHP and PCR test outcome (positive or negative). It was concluded that environmental sampling of the calving area and pre-weaned calf area has potential applications in the assessment of transmission risk on infected farms and could be used to monitor the efficacy of biosecurity measures over time. Further research conducted on a larger scale is required to support these results.

Integrated environmental characterization and assessment of an exposed historic manure repository

Historic manure stockpiled (active from 1935 through 2018) in a repository mound approximately 15 m high with a 31,415.93 m2 footprint was sampled from various depths at six locations in an environmental assessment framework. The manure samples were analyzed for nutrient content to investigate potential application as a soil amendment to local fields in combination with biowaste disposal regulations. Results indicate that manure can be used as a soil amendment; however, different crops and land conditions require specific nutrients, and application must be determined accordingly. Likewise, the manure analysis did not indicate any negative issues that would disallow land application as a disposal option. In addition to limiting environmental soil boring into the manure repository, two-dimensional geophysical electrical resistivity imaging was performed to characterize and quantify the deposited manure. Based on those efforts, the material volume within the site's manure repository was calculated to be 611,942.354 cubic meters (m3). Finally, based on the geophysical results and the historical information about the manure's deposited volume in the study area, an estimation of the released landfill gases and its expected produced energy is presented.

Potential output and risk of commonly administered veterinary antibiotics from small-scale livestock farms to surrounding areas in Northwest China

The concern over antibiotic pollution from animal husbandry has significantly increased over recent years. However, few studies on output and environmental risk of veterinary antibiotics (VAs) throughout different exposure matrices from small-scale livestock farms (SSLFs) have been explored. This study explored the output and environmental risk of three classes of VAs (sulfonamides (SAs), tetracyclines (TCs), fluoroquinolones (FQs)) in three different types of environmental media (manure, soil, and plants/vegetables) derived from four livestock feedlots in the Hexi Corridor of Northwest China. Following, a risk assessment was conducted to identify the hazardous potential of these VAs on the ecological health of the surrounding environment. A total of 108 soil, 36 manure, 12 plants/vegetables, and 15 animal product samples were collected from the animal feedlots for analysis. The results showed that each of the three groups of VAs were detected in the soil, manure and plant samples derived from all four feedlots in varying levels. In the soil samples, the detection rate of SAs (68%) was higher than the TCs (57%) and the FQs (27%). The total concentration of VAs ranged from not detected (n.d.) to 275 ng/g, while chlortetracycline (CTC) was the most abundant (275 ng/g) of the VAs in soil samples. The SAs had the highest detection rate (100%), followed by TCs (89%), and FQs (78%) in manure samples. The total concentration of VAs residues ranged from n. d. to 105 ng/g, of which CTC was as high as 91 ng/g in manures. In the plant/vegetable samples, the TCs had the highest detection rate (58%), while sulfamethazine (SDM) was the most abundant (32 ng/g). The total concentration of the VAs ranged from n. d. to 65 ng/g in the plant/vegetable samples. The target VAs were not detected in animal products. Measurements of the composition of VAs in soil samples at different vertical depths as well as horizontal distances from the manure accumulation sites showed that VAs were partially retained in the soil of the feedlots and were distributed into the surrounding environment both horizontally and vertically. It is suspected that the detected VAs could be accumulated in agricultural soils since they could be found in most of the sampled manures and soils in SSLFs. These results highlighted the necessity of considering SSLF practices to mange the accumulation and disposal of manure mitigating and controlling VA pollution.

High Occurrence of Multiple-Drug Resistance Mediated by Integron in Aeromonas Isolated from Fish-Livestock Integrated Farms

Fish-livestock integrated farming is a traditional practice in South China and Southeast Asia. The administration of antimicrobial agents in livestock may potentially facilitate the spread of antimicrobial resistance to aquaculture through the drainage of livestock manure. Aeromonas spp. are fish pathogens that are the predominant bacteria in water bodies. The aim of this study was to characterize multiple-drug-resistantAeromonas spp. isolated from integrated and nonintegrated fish-livestock farms and to investigate the occurrence of class 1 integrons. Of the 481 Aeromonas strains examined in this study, isolates from pig manure, fish, and environmental samples from pig-fish or goose-fish integrated farms were significantly more resistant to various antimicrobial agents than those from nonintegrated farms P &lt; 0.05 . High rates of resistance to nalidixic acid and sulfamonomethoxine were observed in the isolates from the integrated farm. Fifty Aeromonas isolates (10.4%) from integrated farms contained class 1 integrons. In addition, 96.0% (48/50) of the integron-positive strains displayed multiple-drug resistance. Ten types of gene cassette arrays were determined by sequencing, including dfrA17, dfrA12-orfF-aadA2, catB8, dfrB4-catB3-aadA1, aac6(6′)-Ib-cr-arr-3, aac-II-blaOXA-21-catB3, aar2-aacA4-drfA1-orfC, aac(6′)-Ib-cr, dfrA15, and dfrB4-catB3-blaOXA-10-aadA1. Notably, among the 50 integron-positive isolates, twenty isolates showed integrons located in plasmids, which may facilitate the transmission of resistance in integrated farms. Our investigation confirmed the high prevalence of multiple antimicrobial resistances mediated by class 1 integrons in Aeromonas isolates from integrated farms. Therefore, it is necessary to establish a risk assessment method for antimicrobial resistance in aquaculture.

Characterization of Escherichia coli and Other Enterobacterales Resistant to Extended-Spectrum Cephalosporins Isolated from Dairy Manure in Ontario, Canada.

Extended-spectrum cephalosporins (ESCs) resistance genes, such as blaCTX-M, blaCMY, and blaSHV, have been found regularly in bacteria from livestock. However, information on their distribution in dairy cattle in Canada and on the associated genome sequences of ESC-resistant Enterobacterales is sparse. In this study, the diversity and distribution of ESC-resistant Escherichia coli throughout manure treatments in six farms in Southern Ontario were assessed over a one-year period, and their ESC-resistance plasmids were characterized. The manure samples were enriched using selective media. The resulting isolates were screened via polymerase chain reaction for blaCTX-M, blaCMY, and blaSHV. No E. coli carrying blaSHV were detected. Escherichia coli (n = 248) carrying blaCTX-M or blaCMY underwent whole-genome sequencing using an Illumina MiSeq/NextSeq. These isolates were typed using multilocus sequence typing (MLST) and their resistance gene profiles. A subset of E. coli (n = 28) were sequenced using Oxford Nanopore Technologies. Plasmids were assembled using Unicycler and characterized via the resistance genes pattern, replicon type, plasmid MLST, phylogenetic analysis, and Mauve alignments. The recovery of ESC-resistant Enterobacterales (18 species, 8 genera) was drastically reduced in manure outputs. However, multiple treatment stages were needed to attain a significant reduction. 62 sequence types were identified, with ST10, ST46, ST58, ST155, ST190, ST398, ST685, and ST8761 being detected throughout the treatment pipeline. These STs overlapped with those found on multiple farms. The ESC-resistance determinants included CTX-M-1, -14, -15, -17, -24, -32, -55, and CMY-2. The plasmids carrying blaCTX-M were more diverse than were the plasmids carrying blaCMY. Known "epidemic plasmids" were detected for both blaCTX-M and blaCMY. IMPORTANCE The increase in antimicrobial resistance is of concern for human and animal health, especially when resistance is conferred to extended-spectrum cephalosporins, which are used to treat serious infections in both human and veterinary medicine. Bacteria carrying extended-spectrum cephalosporin resistance genes, including blaCTX-M and blaCMY, are frequently found in dairy manure. Manure treatment influences the loads and diversity of bacteria, including those carrying antimicrobial resistance genes, such as Enterobacterales and Escherichia coli. Any bacteria that survive the treatment process are subsequently applied to the environment. Enterobacterales carrying blaCTX-M or blaCMY can contaminate soil and crops consumed by humans and animals, thereby increasing the potential for antimicrobial resistance genes to integrate into the human gut microflora through horizontal gene transfer. This furthers the dissemination of resistance. Therefore, it is imperative to understand the effects manure treatments have on ESC-resistance in environmentally applied manure.

Characteristics and source analysis of polycyclic aromatic hydrocarbons in organisms and manure near Ardley Island, Antarctica

In order to better understand the migration and accumulation behavior of polycyclic aromatic hydrocarbons (PAHs) in biological chains in the cold environment around the Ardley Island, a variety of biological and penguin manure samples during China's 36th Antarctic Scientific expedition have been collected and studied. A certain difference in PAHs concentration was observed in the environmental samples, and the order of size was as follows: fish > limpets > krill > manure > brown alga > mosses. The percentage of PAHs with different ring numbers in brown alga, moss, and krill was in the following order: three rings > two rings > four rings > five rings > six rings. The proportion of HMW-PAHs in limpets and fish samples was higher than that in brown alga, moss, manure, and krill samples. The main source of PAHs in environmental samples near Ardley Island is oil, which may be due to the development of tourism in Antarctica, the number of ships and human activities around the region. Therefore, it is imperative to strengthen the protection of the ecological environment in the area around Ardley Island.

Bioaugmentation removal and microbiome analysis of the synthetic estrogen 17α-ethynylestradiol from hostile conditions and environmental samples by Pseudomonas citronellolis SJTE-3

Synthetic estrogens are emerging environmental contaminants with great estrogenic activities and stable structures that are widespread in various ecological systems and significantly threaten the health of organisms. Pseudomonas citronellolis SJTE-3 is reported to degrade the synthetic estrogen 17α-ethynylestradiol (EE2) efficiently in laboratory conditions. In this work, the environmental adaptability, the EE2-degrading properties, and the ecological effects of P. citronellolis SJTE-3 under different hostile conditions (heavy metals and surfactants) and various natural environment samples (solid soil, lake water, and pig manure) were studied. Strain SJTE-3 can tolerate high concentrations of Zn2+ and Cr3+, but is relatively sensitive to Cu2+. Tween 80 of low concentration can significantly promote EE2 degradation by strain SJTE-3, different from the repressing effect of Triton X-100. High concentration of Tween 80 prolonged the lagging phase of EE2-degrading process, while the final EE2 removal efficiency was improved. More importantly, strain SJTE-3 can grow normally and degrade estrogen stably in various environmental samples. Inoculation of strain SJTE-3 removed the intrinsic synthetic and natural estrogens (EE2 and estrone) in lake water samples in 4 days, and eliminated over 90% of the amended 1 mg/L EE2 in 2 days. Bioaugmentation of strain SJTE-3 in EE2-supplied solid soil and pig manure samples achieved a removal rate of over 55% and 70% of 1 mg/kg EE2 within 2 weeks. Notably, the bioaugmentation of extrinsic strain SJTE-3 had a slight influence on indigenous bacterial community in pig manure samples, and its relative abundance decreased significantly after EE2 removal. Amendment of EE2 or strain SJTE-3 in manure samples enhanced the abundance of Proteobacteria and Actinobacteria, implying their potential in utilizing EE2 or its metabolites. These findings not only shed a light on the environment adaptability and degradation efficiency of strain SJTE-3, but also provide insights for bioremediation application in complex and synthetic estrogen polluted environments.

Evaluation of Bacterial Composition and Viability of Equine Feces after Processing for Transplantation

Fecal microbiota transplantation (FMT) has been used empirically for decades in equine medicine to treat intestinal dysbiosis but evidence-based information is scarce. This in vitro study aimed at assessing the effect of a commonly used pre-FMT processing method on the bacterial composition and viability of the fecal filtrate. Three samples of fresh equine manure (T0) were processed identically: the initial manure was mixed with 1 L of lukewarm water and chopped using an immersion blender to obtain a mixture (T1), which was left uncovered during 30 min (T2) and percolated through a sieve to obtain a fecal filtrate (T3). Samples were taken throughout the procedure (Tn) and immediately stored at 4 °C until processing. The 16S rDNA amplicon profiling associated with propidium monoazide treatment was performed on each sample to select live bacteria. Analyses of α and β diversity and main bacterial populations and quantitative (qPCR) analysis were performed and statistically compared (significance p < 0.05) between time points (T0-T3). No significant differences in ecological indices or mean estimated total living bacteria were found in the final fecal filtrate (T3) in regard to the original manure (T0); however, relative abundances of some minor genera (Fibrobacter, WCHB1-41_ge and Akkermansia) were significantly different in the final filtrate. In conclusion, the results support the viability of the major bacterial populations in equine feces when using the described pre-FMT protocol.