Probing the cw-Laser-Induced Fluorescence Enhancement in CsPbBr3 Nanocrystal Thin Films: An Interplay between Photo and Thermal Activation.

Perovskite nanocrystals hold significant promise for a wide range of applications, including solar cells, LEDs, photocatalysts, humidity and temperature sensors, memory devices, and low-cost photodetectors. Such technological potential stems from their exceptional quantum efficiency and charge carrier conduction capability. Nevertheless, the underlying mechanisms of photoexcitation, such as phase segregation, annealing, and ionic diffusion, remain insufficiently understood. In this context, we harnessed hyperspectral fluorescence microspectroscopy to advance our comprehension of fluorescence enhancement triggered by UV continuous-wave (cw) laser irradiation of CsPbBr3 colloidal nanocrystal thin films. Initially, we explored the kinetics of fluorescence enhancement and observed that its efficiency (φph) correlates with the laser power (P), following the relationship φph = 7.7⟨P⟩0.47±0.02. Subsequently, we estimated the local temperature induced by the laser, utilizing the finite-difference method framework, and calculated the activation energy (Ea) required for fluorescence enhancement to occur. Our findings revealed a very low activation energy, Ea ∼ 9 kJ/mol. Moreover, we mapped the fluorescence photoenhancement by spatial scanning and real-time static mode to determine its microscale length. Below a laser power of 60 µW, the photothermal diffusion length exhibited nearly constant values of approximately (22 ± 5) µm, while a significant increase was observed at higher laser power levels. These results were ascribed to the formation of nanocrystal superclusters within the film, which involves the interparticle spacing reduction, creating the so-called quantum dot solid configuration along with laser-induced annealing for higher laser powers.

WGS of intrauterine E. coli from cows with early postpartum uterine infection reveals a non-uterine specific genotype and virulence factors.

Escherichia coli has been attributed to playing a major role in a cascade of events that affect the prevalence and severity of uterine disease in cattle. The objectives of this project were to (i) define the association between the prevalence of specific antimicrobial resistance and virulence factor genes in E. coli with the clinical status related to uterine infection, (ii) identify the genetic relationship between E. coli isolates from cows with diarrhea, with mastitis, and with and without metritis, and (iii) determine the association between the phenotypic and genotypic antimicrobial resistance identified on the E. coli isolated from postpartum cattle. Bacterial isolates (n = 148) were obtained from a larger cross-sectional study. Cows were categorized into one of three clinical groups before enrollment: metritis, cows with purulent discharge, and control cows. For genomic comparison, public genomes (n = 130) from cows with diarrhea, mastitis, and metritis were included in a genome-wide association study, to evaluate differences between the drug classes or the virulence factor category among clinical groups. A distinct E. coli genotype associated with metritis could not be identified. Instead, a high genetic diversity among the isolates from uterine sources was present. A virulence factor previously associated with metritis (fimH) using PCR was not associated with metritis. There was moderate accuracy for whole-genome sequencing to predict phenotypic resistance, which varied depending on the antimicrobial tested. Findings from this study contradict the traditional pathotype classification and the unique intrauterine E. coli genotype associated with metritis in dairy cows.IMPORTANCEMetritis is a common infectious disease in dairy cattle and the second most common reason for treating a cow with antimicrobials. The pathophysiology of the disease is complex and is not completely understood. Specific endometrial pathogenic Escherichia coli have been reported to be adapted to the endometrium and sometimes lead to uterine disease. Unfortunately, the specific genomic details of the endometrial-adapted isolates have not been investigated using enough genomes to represent the genomic diversity of this organism to identify specific virulence genes that are consistently associated with disease development and severity. Results from this study provide key microbial ecological advances by elucidating and challenging accepted concepts for the role of Intrauterine E. coli in metritis in dairy cattle, especially contradicting the existence of a unique intrauterine E. coli genotype associated with metritis in dairy cows, which was not found in our study.

Agreement among deep nasopharyngeal sampling culture results for 3 different swab types in preweaning dairy calves.

Accurate isolation and identification of pathogens for an animal with bovine respiratory disease are of critical importance to direct appropriate decision-making related to the treatment of individual animals, as well as control and prevention options in a herd setting. The objective of this study was to compare nasopharyngeal sampling approaches to evaluate accuracy and agreement for the recovery of Mannheimia haemolytica (MH) and Pasteurella multocida (PM) from deep nasopharyngeal swabs (DNS) using 3 different swabs. Deep nasopharyngeal samples were collected from 45 dairy calves using 3 swabs: (1) double-guarded culture swab (DGS); (2) single-guarded culture swab (SGS); and (3) unguarded culture swab (UGS). To evaluate the degree of agreement between DGS, SGS, and UGS, culture results were compared for each calf sampled by using a kappa agreement test. Overall, findings from our study support that when using either SGS or DGS for DNS sampling of preweaning calves, a high agreement for recovery of PM is observed. A low recovery of MH was observed in the study, limiting the conclusion comparing the 3 DNS methods. Use of UGS is considered a potential alternative; however, a higher percentage of polymicrobial growth was found with UGS samples.

Antimicrobial resistance of Escherichia coli from dairy farms participating in an antimicrobial stewardship educational program for farm employees.

Antimicrobial use in food-producing animals is under increasing scrutiny due to the potential effect on the selection of antimicrobial-resistant bacteria that may be transmitted to humans by direct contact, with the food chain, or the environment. Novel data monitoring commensal Escherichia coli from dairy farms is essential for understanding antimicrobial resistance (AMR) patterns and their association with herd health management practices. The objectives of this study were to: (1) compare the prevalence of antimicrobial resistance in the E. coli isolates from the hospital, fresh, and mid-lactation pens from 18 conventional dairy farms participating in an educational training program in antimicrobial stewardship practices in California and Ohio, and (2) to characterize the prevalence of antimicrobial resistance of commensal E. coli isolated from pooled fecal pat samples before and 3 mo after participating in the educational training program. Pooled fecal pat samples were collected from the hospital pen, the fresh pen (1 to 5 DIM), and the mid-lactation pens (90 to 150 DIM) on conventional dairies in CA (n = 9) and OH (n = 9). Fecal samples were collected as part of a larger study using a quasi-experimental design that assigned farms to the training intervention group (TG; 6 per state) or the control group (CG; 3 per state). For the TG, farm worker(s) identified as having the task of diagnosis and treatment of adult cows on the farm participated in a training program on antimicrobial stewardship practices. Pooled fecal samples (n = 7) were collected at enrollment and 3 mo after completing the intervention on each of the participating farms (n = 18), followed by culture for E. coli isolation and antimicrobial sensitivity testing using the broth microdilution methodology. Logistic regression models were used to evaluate the association between E. coli antimicrobial resistance patterns with the training intervention and farm-level factors. No effect was observed in the prevalence of resistant isolates between the control and intervention farms after the training was delivered. Isolates from the hospital pens were 2.48 (95% CI: 1.06-6.22, P = 0.03) and 5.61 (95% CI: 1.94-16.91, P < 0.001) times, more likely to be resistant to streptomycin and chloramphenicol, respectively, than isolates from the mid-lactation pens. Our findings indicate there was a higher prevalence of AMR in E. coli associated with the hospital pen within the farm, while the training program for 3 mo did not affect the prevalence of AMR in E. coli on the farms participating in the program. Further research efforts should be conducted to identify factors driving AMR at the pen level, as well as approaches that could be used to reduce the risk of disseminating AMR from sick pens to animals being housed and to other pens on the farm.

Unveiling the microbiome during post-partum uterine infection: a deep shotgun sequencing approach to characterize the dairy cow uterine microbiome.

BACKGROUND: The goal of this study was to assess the microbial ecology and diversity present in the uterus of post-partum dairy cows with and without metritis from 24 commercial California dairy farms using shotgun metagenomics. A set subset of 95 intrauterine swab samples, taken from a larger selection of 307 individual cow samples previously collected, were examined for α and ß diversity and differential abundance associated with metritis. Cows within 21 days post-partum were categorized into one of three clinical groups during sample collection: control (CT, n = 32), defined as cows with either no vaginal discharge or a clear, non-purulent mucus vaginal discharge; metritis (MET, n = 33), defined as a cow with watery, red or brown colored, and fetid vaginal discharge; and purulent discharge cows (PUS, n = 31), defined as a non-fetid purulent or mucopurulent vaginal discharge. RESULTS: All three clinical groups (CT, MET, and PUS) were highly diverse, with the top 12 most abundant genera accounting for 10.3%, 8.8%, and 10.1% of mean relative abundance, respectively. The α diversity indices revealed a lower diversity from samples collected from MET and PUS when compared to CT cows. PERMANOVA statistical testing revealed a significant difference (P adjusted < 0.01) in the diversity of genera between CT and MET samples (R2 = 0.112, P = 0.003) and a non-significant difference between MET and PUS samples (R2 = 0.036, P = 0.046). ANCOM-BC analysis revealed that from the top 12 most abundant genera, seven genera were increased in the natural log fold change (LFC) of abundance in MET when compared to CT samples: Bacteroides, Clostridium, Fusobacterium, Phocaeicola, Porphyromonas, Prevotella, and Streptococcus. Two genera, Dietzia and Microbacterium, were decreased in natural LFC of abundance when comparing MET (regardless of treatment) and CT, while no changes in natural LFC of abundance were observed for Escherichia, Histophilus, and Trueperella. CONCLUSIONS: The results presented here, are the current deepest shotgun metagenomic analyses conducted on the bovine uterine microbiome to date (mean of 256,425 genus-level reads per sample). Our findings support that uterine samples from cows without metritis (CT) had increased α-diversity but decreased ß-diversity when compared to metritis or PUS cows, characteristic of dysbiosis. In summary, our findings highlight that MET cows have an increased abundance of Bacteroides, Porphyromonas, and Fusobacterium when compared to CT and PUS, and support the need for further studies to better understand their potential causal role in metritis pathogenesis.

Changes in Uterine Metabolome Associated with Metritis Development and Cure in Lactating Holstein Cows.

The objective of this study was to identify alterations in the vaginal discharge (VD) metabolome and potential biomarkers to predict metritis development and a cure in dairy cows. This prospective cohort study was conducted on two dairies located in CA and TX. Vaginal discharge was evaluated and collected using the Metricheck® device. Cows were examined for metritis at 4, 7, and 9 days in milk (DIM). Cows with a fetid, watery, and reddish-brown uterine discharge were classified as having metritis and randomized to receive ceftiofur (n = 10) or remain untreated (n = 7). A cure was defined as the absence of a fetid, watery, reddish-brown uterine discharge at 14 d after enrollment. Vaginal discharge samples were collected from 86 cows within 6 h after parturition, at 4 and 7 DIM, at metritis diagnosis, and at 4 and 7 days after metritis diagnosis. Cows with metritis (MET; n = 17) were paired with counterparts without metritis (HTH) of a similar DIM and parity (n = 34). The uterine metabolome was evaluated using untargeted gas chromatography time-of-flight mass spectrometry (GC-TOF-MS). Metabolomic data were analyzed using the MetaboAnalyst 5.0. Data were log-transformed and auto-scaled for normalization. Univariate analyses, including the fold-change, were performed to identify the metabolites linked to metritis development and its cure and principal component analysis and partial least squares discriminant analysis were performed to explain metabolite variance between animals developing or not developing metritis and being cured or not being cured of metritis. Comparing HTH with MET cows at calving, 12 metabolites were upregulated, and one was downregulated. At four and seven DIM, 51 and 74 metabolites, respectively, were altered between MET and HTH cows. After metritis development, three and five metabolites were upregulated in cows that were cured and in cows that received treatment and were cured, respectively. In all scenarios, the metabolites lignoceric, malic, and maleic acids, ornithine, and hypotaurine, which are associated with arginine/aminoacyl-tRNA biosynthesis and taurine/purine metabolism, were upregulated in HTH cows. Metritis was associated with changes in the uterine metabolome. Cows not being cured of metritis had changes in the uterus metabolome independent of receiving ceftiofur or remaining untreated. Metabolome analysis may be an important tool to understand the vaginal discharge changes during postpartum and the dynamics of metritis development and cures and help to identify biomarkers to predict metritis being cured.

Physicochemical properties of ultrasmall colloidal silver nanoparticles: an experimental and computational approach.

Silver nanoparticles (AgNPs) exhibit very interesting properties that have been employed in several kinds of applications spanning from antibacterial activity to plasmon-polaritons generation. Nanoparticle size strongly influences these applications. However, the characterization of ultrasmall AgNPs (particle diameter < 10 nm) encompassing different aspects such as average size, polydispersion, shape (anisotropy), concentration, and density remains a challenging task. To address these challenges, we combined TEM measurements with a computational framework based on Mie-Gans theory. This allowed us to describe the aforementioned AgNP features accurately. The synthesis of AgNPs in an aqueous medium involved the use of silver nitrate as a chemical precursor and sodium borohydride as a reducing agent, with polyvinylpyrrolidone acting as a stabilizing agent. Our outcomes showed that increasing the concentration of the precursor and reducing agent with a fixed 1 : 2 molar ratio tends to yield ultrasmall AgNPs with low to moderate polydispersion, a nearly spherical shape (low anisotropy), concentration in the nanomolar range and density close to silver bulk. Also, we established an analytical expression that correlates the extinction molar absorptivity to AgNP size considering the nanoparticle shape. Notably, the computational framework proved to be highly effective in extracting crucial information about the AgNPs from UV-vis spectroscopy data. In conclusion, our study sheds light on the unique properties of ultrasmall AgNPs and presents a comprehensive approach for properly characterizing these nanoparticles, paving the way for further advancements in their applications.

In vitro evaluation of the effect of transport medium, temperature, and time on the recovery of Mannheimia haemolytica and Pasteurella multocida.

Appropriate sample collection, storage conditions, and time for transport to the laboratory are important for an accurate diagnostic result. We evaluated the effects of transport storage medium type, time of storage, and storage temperatures on Mannheimia haemolytica (MH) and Pasteurella multocida (PM) recovery using an in vitro model simulation. A quantitative culture method, using colony-forming units per milliliter, was used to recover MH or PM by an in vitro model with cotton swabs. Three independent trials were conducted, in which cotton swabs were inoculated with MH or PM and placed in either (1) a sterile 15-mL polypropylene tube without transport medium (dry), (2) Amies culture medium with charcoal (ACM), or (3) Cary-Blair transport agar (CBA). Swabs were evaluated for recovery of MH or PM when stored at 3 temperatures (4°C, 23°C, or 36°C) and after storage for 8 h, 24 h, or 48 h. From all study group combinations, a total of 162 individual independent swabs were evaluated. The nonparametric Dunn all-pairs approach was used to compare the proportion of culturable bacteria, between the various storage media, temperature, and time point combinations. The proportion of MH in samples stored at 4°C was significantly higher for ACM and CBA than dry storage at 24 and 48 h. The MH samples stored at 36°C had a significantly higher proportion for ACM and CBA than dry storage at 24 h. The proportion of PM in samples stored at 4°C was significantly lower for ACM compared with dry at 8 h but significantly higher at 48 h. The PM samples stored at 23°C in ACM had a significantly higher proportion than dry samples at 24 h, and, at 48 h, ACM and CBA had a significantly higher proportion than the dry group. All swabs stored at 36°C for 48 h had a proportion close to zero, indicating decreasing diagnostic efficacy. These results support the use of transport media such as ACM and CBA for increasing the detection of PM and MH from samples, especially when samples are exposed to high temperatures. The combination of longer periods from collection of samples to diagnostic evaluation (>24 h) and higher storage temperatures (>23°C) were shown to significantly impair diagnostic accuracy.

Antimicrobial stewardship on the dairy: Evaluating an on-farm framework for training farmworkers.

Dairy farmworkers are commonly responsible for disease diagnosis and routine treatment decisions for cattle. This highlights the importance of farmworkers' knowledge and skills to successfully implement judicious use of antimicrobials in livestock production systems. The main objectives of this project were to develop and evaluate an on-farm educational program for farmworkers in antimicrobial stewardship in adult dairy cattle. A longitudinal quasi-experimental study design was used, by enrolling 12 conventional dairy farms in the United States (6 in California and 6 in Ohio). Farmworkers responsible for treatment decisions on the farm (n = 25) participated in a didactic and hands-on 12-wk antimicrobial stewardship training program led by the investigators. All antimicrobial stewardship training materials were available in Spanish and English. Interactive short videos with audio were developed to cover the learning objectives for each of the 6 teaching modules: antimicrobial resistance, treatment protocols, visual identification of sick animals, clinical mastitis, puerperal metritis, and lameness. Pre- and post-training assessments were administered using an online training assessment tool to evaluate changes in knowledge and attitudes about antimicrobial stewardship practices. Cluster analysis and multiple correspondence analyses were conducted to evaluate the association among categorical variables for participants' level of change in knowledge and its association with language, farm size, and state. A 32% average increase in knowledge was observed through an assessment conducted after completing the antimicrobial stewardship training, compared with the pre-training assessment. A significant improvement in 7 of 13 attitude questions related to antimicrobial stewardship practices on the farm was observed. Knowledge and attitude scores of participants on antimicrobial stewardship and identification of sick animals significantly improved after completing the antimicrobial stewardship training. The results observed in this study support the relevance of antimicrobial stewardship training programs targeting farmworkers to improve antimicrobial drug use knowledge and skills.

Prevalence of antimicrobial resistance in fecal Escherichia coli and Enterococcus spp. isolates from beef cow-calf operations in northern California and associations with farm practices.

Antimicrobials are necessary for the treatment of bacterial infections in animals, but increased antimicrobial resistance (AMR) is becoming a concern for veterinarians and livestock producers. This cross-sectional study was conducted on cow-calf operations in northern California to assess prevalence of AMR in Escherichia coli and Enterococcus spp. shed in feces of beef cattle of different life stages, breeds, and past antimicrobial exposures and to evaluate if any significant factors could be identified that are associated with AMR status of the isolates. A total of 244 E. coli and 238 Enterococcus isolates were obtained from cow and calf fecal samples, tested for susceptibility to 19 antimicrobials, and classified as resistant or non-susceptible to the antimicrobials for which breakpoints were available. For E. coli, percent of resistant isolates by antimicrobial were as follows: ampicillin 100% (244/244), sulfadimethoxine 25.4% (62/244), trimethoprim-sulfamethoxazole 4.9% (12/244), and ceftiofur 0.4% (1/244) while percent of non-susceptible isolates by antimicrobial were: tetracycline 13.1% (32/244), and florfenicol 19.3% (47/244). For Enterococcus spp., percent of resistant isolates by antimicrobial were as follows: ampicillin 0.4% (1/238) while percent of non-susceptible isolates by antimicrobial were tetracycline 12.6% (30/238) and penicillin 1.7% (4/238). No animal level or farm level management practices, including antimicrobial exposures, were significantly associated with differences in isolate resistant or non-susceptible status for either E. coli or Enterococcus isolates. This is contrary to the suggestion that administration of antibiotics is solely responsible for development of AMR in exposed bacteria and demonstrates that there are other factors involved, either not captured in this study or not currently well understood. In addition, the overall use of antimicrobials in this cow-calf study was lower than other sectors of the livestock industry. Limited information is available on cow-calf AMR from fecal bacteria, and the results of this study serve as a reference for future studies to support a better understanding and estimation of drivers and trends for AMR in cow-calf operations.

Salmonella enterica Serovar Dublin from Cattle in California from 1993 to 2019: Characterization and Analysis of Antimicrobial Resistance Diversity.

For this study, antimicrobial susceptibility data for Salmonella enterica subsp. enterica serovar Dublin (S. Dublin)-a well-known cattle-adapted pathogen with current concerns for multidrug resistance-were recovered from cattle at the California Animal Health and Food Safety Laboratory System (CAHFS) over the last three decades (1993-2019) and were evaluated using tools to capture diversity in antimicrobial resistance. For this purpose, minimum inhibitory concentration (MIC) testing was conducted for 247 clinical S. Dublin isolates. Antimicrobial resistance (AMR) profiles revealed a predominant core multidrug-resistant pattern in the three most common AMR profiles observed. Antimicrobial resistance richness, diversity, and similarity analysis revealed patterns for changes in AMR profiles for different age groups. Discriminant analysis using MIC log2-transformed data revealed changes in MIC for year groups, with a time-sequence pattern observed. Drivers for reduced susceptibility were observed for 3rd generation cephalosporins and quinolones observed for more recent year groups (2011-2015 and 2016-2019) when compared to older year groups (1993-1999 and 2000-2005). Together, these results highlight the changes in the diversity of AMR profiles, as well as changes in susceptibility of S. Dublin over time for critical antimicrobials of importance to both animals and humans, and support the need for continued monitoring and efforts that will support judicious use of antimicrobials, especially for these two drug classes.

Assessment of the knowledge and behavior of backyard and small-scale producers in California regarding disease prevention, biosecurity practices and antibiotics use.

The number and popularity of backyard poultry and livestock farming have rapidly increased in California as well as other states in the United States following consumers' preference for local and organic products in the last few years. This study aimed to investigate current on-farm management and farmers' understanding of Veterinary Feed Directive (VFD) and California Senate Bill (SB) 27 implications for disease prevention, biosecurity procedures, and antimicrobial use in small-scale and backyard farms in California. The survey consisted of 38 questions. The responses of 242 backyard and small-scale livestock owners were investigated in this study. Descriptive statistics summarized survey responses, and multivariable logistic regression evaluated the association of antibiotics purchase and use, and the impact of VFD and SB27 on antibiotic use with demographics and on-farm management. Backyard and small-scale farmers in California mostly raised chickens or small ruminants with small herd sizes kept for personal use. Antibiotics were generally used for individual treatment of a sick animal with the guidance of a veterinarian. VFD and SB27 implementation promoted the judicious use of antibiotics, specifically, by enhancing the relationship between backyard and small-scale farmers with veterinarians and treating fewer animals with antibiotics under veterinary oversight. Therefore, better access to veterinary service in backyard and small-scale farms will improve the farmer's knowledge of good husbandry practices with judicious antimicrobial use in livestock and finally contribute to reducing the risk of antimicrobial resistance in California.

Survey of Management Practices and Farmers' Perceptions of Diseases on Organic Dairy Cattle Farms in California.

In 2019, California accounted for approximately 40% of organic products in the US, and dairy products and milk were the top organic commodity in the state. The objective of this study was to describe organic dairy cattle farmers' management practices and perceptions of diseases in California. A survey inquiring about farm history and demographics, animal diseases, parasite problems, housing and pasture management, and organic education, was mailed to 160 organic dairy farms, of which 36 (22.5%) responded. Among respondents, the majority (83.9%) were located in Northern California; median farm size was 310 cows, and the dominant breed was Holstein (60.0%). Respondents were more likely to report mastitis in cows (45.2%), pinkeye in heifers (31.3%), and digestive problems in calves (47.0%), as issues affecting their stock "often" or "almost always" in the last 12 months. Although most farmers vaccinated their cattle against Bovine Viral Diarrhea (86.1%) and Escherichia coli mastitis (80.6%), they still described that these diseases frequently impacted their animals. Over half of the farmers did not perceive gastrointestinal parasites or biting flies to be a problem and did not observe signs of lice and mites. According to the results, the management of disease in all age classes is a concern; options and efficacies of alternative therapeutic methods, as well as preventive measures for organic dairies need to be further explored.

Comparison of florfenicol depletion in dairy goat milk using ultra-performance liquid chromatography with tandem mass spectrometry and a commercial on-farm test.

Florfenicol is a broad-spectrum antibiotic commonly prescribed in an extra-label manner for treating meat and dairy goats. Scientific data in support of a milk withdrawal interval recommendation is limited to plasma pharmacokinetic data and minimal milk residue data that is limited to cattle. Therefore, a rapid residue detection test (RRDT) could be a useful resource to determine if milk samples are free of drug residues and acceptable for sale. This study compared a commercially available RRDT (Charm® FLT strips) to detect florfenicol residues in fresh milk samples from healthy adult dairy breed goats treated with florfenicol (40 mg/kg subcutaneously twice 4 days apart) with quantitative analysis of florfenicol concentrations using ultra-performance liquid chromatography with tandem mass spectrometry (UPLC-MS/MS). In addition, storage claims for testing bovine milk using the RRDT were assessed using stored goat milk samples. Milk samples were collected every 12 h for a minimum of 26 days. Commercial RRDT strips remained positive in individual goats ranging from 528 to 792 h (22-33 days) after the second dose, whereas, UPLC-MS/MS indicated the last detectable florfenicol concentration in milk samples ranged from 504 to 720 h (21-30 days) after the second dose. Results from stored milk samples from treated goats indicate that samples can be stored for up to 5 days in the refrigerator and 60 days in the freezer after milking prior to being tested with a low risk of false-negative test results due to drug degradation. Elevated somatic cell counts and bacterial colony were noted in some of the milk samples in this study, but further study is required to understand the impact of these quality factors on RRDT results.

Evaluation of antimicrobial resistance and risk factors for recovery of intrauterine Escherichia coli from cows with metritis on California commercial dairy farms.

The goals of this study were to evaluate factors affecting recovery and antimicrobial resistance (AMR) in intrauterine E. coli in post-partum dairy cows with and without metritis from commercial California dairy farms. Using a cross-sectional study design, a total of 307 cows were sampled from 25 farms throughout California, from which a total of 162 intrauterine E. coli isolates were recovered. During farm visits, cows within 21 days post-partum were categorized in one of three clinical presentation groups before enrollment: metritis (MET, n = 86), defined as a cow with watery, red or brown colored, and fetid vaginal discharge; cows with purulent discharge (PUS, n = 106), defined as a non-fetid purulent or mucopurulent vaginal discharge; and control cows, (CTL, n = 115) defined as cows with either no vaginal discharge or a clear, non-purulent mucus vaginal discharge. Cows diagnosed as MET had significantly higher odds for recovery of E. coli compared to cows diagnosed as CTL (OR = 2.16, 95% CI: 1.17-3.96), with no significant difference observed between PUS and CTL, and PUS and MET. An increase in days in milk (DIM) at the time of sampling was significantly associated with a decrease in the odds ratio for E. coli recovery from intrauterine swabs (OR = 0.94, 95% CI: 0.89-0.98). All intrauterine E. coli were resistant to ampicillin (AMP), with an AMR prevalence of 30.2% and 33.9% observed for chlortetracycline and oxytetracycline, respectively. Only 8.6% of isolates were resistant to ceftiofur (CEFT), one of the most common drugs used to treat cows on farms sampled. No significant difference in the prevalence of AMR was observed among clinical groups at the individual cow level. At the farm level, a significantly higher odds for isolating intrauterine E. coli resistant to chlortetracycline (OR: 2.6; 95% CI: 3.7-58.0) or oxytetracycline (OR: 1.9; 95% CI: 1.4-33.8) was observed at farms that used an intrauterine infusion of oxytetracycline as a treatment for metritis when compared to those farms that did not use this practice. Findings from this study indicate the need for further research supporting a broader understanding of farm practices driving AMR in cows with metritis, as well as data to increase the accuracy of breakpoints for AMR classification of intrauterine E. coli from cattle.

Salmonella enterica Serovar Dublin from Cattle in California from 1993-2019: Antimicrobial Resistance Trends of Clinical Relevance.

Salmonella enterica subsp. enterica serovar Dublin (S.Dublin) is a cattle-adapted pathogen that has emerged as one of the most commonly isolated and multidrug resistant (MDR) serovars in cattle. S.Dublin may be shed in feces, milk, and colostrum and persist in asymptomatic cattle, leading to spread and outbreaks in herds. Though infections with S.Dublin in humans are rare, they are frequently severe, with extraintestinal spread that requires hospitalization and antimicrobial therapy. To determine minimum inhibitory concentration (MIC) and antimicrobial resistance (AMR) patterns and trends in cattle in California, broth microdilution testing was performed on 247 clinical S. Dublin isolates recovered from cattle at the California Animal Health and Food Safety Laboratory System (CAHFS) over the last three decades (1993-2019). Mean MICs and classification of resistance to antimicrobial drugs using a clinical livestock panel and the National Antimicrobial Resistance Monitoring System (NARMS) Gram-negative drug panels were utilized to assess prevalence and trends in AMR. Findings indicate an increase in AMR for the years 1993 to 2015. Notably, compared to the baseline year interval (1993-1999), there was an increase in resistance among quinolone and cephalosporin drugs, as well as an increased number of isolates with an MDR profile.

Effect of Intramammary Dry Cow Antimicrobial Treatment on Fresh Cow's Milk Microbiota in California Commercial Dairies.

This study used 16S rRNA sequencing to evaluate the effects of dry cow antimicrobial therapy on the udder milk microbiota by comparing the microbial populations in milk at dry-off (DRY) (~60 days before calving) and post-partum (FRESH) (4-11 days after calving) from cows receiving an intramammary antibiotic infusion prior to dry-off (IMT) and cows that did not receive treatment (CTL). Milk was collected from 23 cows from the IMT group and 27 cows from the CTL group. IMT and DRY samples had a greater correlation with the genera Brevibacterium and Amaricoccus, and the family Micrococcaceae, when compared to IMT and FRESH samples. CTL group samples collected at DRY had a greater correlation with the genera Akkermansia and Syntrophus, when compared to FRESH samples; no bacterial taxa were observed to have a significant correlation with FRESH samples in the CTL group. DRY samples collected from the CTL group had a greater correlation with the genus Mogibacterium when compared to IMT and CTL samples. For DRY samples collected from the IMT group, a greater correlation with the genus Alkalibacterium when compared to DRY and CTL samples, was observed. The lack of a correlation for FRESH samples between the CTL and IMT treatment groups indicated that intramammary antimicrobial dry cow therapy had no significant effect on the udder milk microbiota post-partum.

Defining clinical diagnosis and treatment of puerperal metritis in dairy cows: A scoping review.

Although puerperal metritis (PM) is a common infectious disease in dairy cattle, there are currently discrepancies between clinical case definitions within and between available peer-reviewed literature and on-farms practices. The inconsistent use of PM criteria across studies and on-farms practices can result in disparities related to recommendations for treating cows, affecting judicious use of antimicrobials. The objective of this study was to systematically review the peer-reviewed literature for clinical signs used for case definition of PM. The criteria used included local (e.g., vaginal discharge) and systemic clinical signs of infection (e.g., fever, drop in milk). The Preferred Reporting Items for Systematic Review and Meta-Analysis extension for scoping reviews protocols were used to screen commonly used databases. Following this protocol, one reviewer screened title and abstract for eligibility (n = 2,096), followed by full-text screening of selected articles (n = 396) by 2 reviewers to confirm eligible articles (n = 174). The most frequently cited reference article (37.5%) for the definition of PM was published in 2006, followed by articles published between 1998 and 2009 (13%). In 40.2% of articles, no reference was provided for definition of PM; vaginal discharge was described in terms of color, odor, and viscosity when related to the PM definition. Terms used for description of vaginal discharge color were red-brown (61.4%), red (5.1%), brown (8.6%), chocolate (4%), white (1.7%), yellow (0.5%), pink (5.7%), or gray (0.5%); vaginal discharge color was not reported in 24.1% articles. The vaginal discharge odor was described as fetid (75.8%), putrid (5.1%), foul (10.3%), or other (5.7%; e.g., abnormal, malodorous, odoriferous); odor was not mentioned in 7.4% of articles. The vaginal discharge viscosity was described as watery (74.1%), purulent (27%), mucopurulent (8.6%), thin (4%), serous (2.8%), or abnormal (2.3%) and was not mentioned in 11.5% of articles. Fever was included in 59.7% of articles as a criterion for PM diagnosis. The most used rectal temperature threshold was ≥39.5°C (56.8%), followed by ≥39.2°C (2.8%). Approaches used for vaginal discharge evaluation included rectal palpation (37.3%), intravaginal exploration with a gloved hand (18.4%), Metricheck (9.8%), or speculum (5.7%); and in 28.7% of articles, diagnostic tools used were not mentioned. Many of the color and odor vaginal discharge descriptions observed in the literature, used synonymous words to describe the same vaginal discharge sample, highlighting a lack of terminology consensus that could result in disagreements, especially due to the subjective character of these clinical evaluations of vaginal discharge color and odor. Although select consensus articles are available, it is common for studies to disregard a reference when defining PM cases. Furthermore, our findings highlight the need for a robust and clear consensus on criteria and terminology used to diagnose PM.

Estimating the Rates of Acquisition and loss of Resistance of Enterobacteriaceae to Antimicrobial Drugs in Pre-Weaned Dairy Calves.

The objective of this study was to investigate the effect of the antimicrobial drugs (AMD) on the shedding of resistant Enterobacteriaceae in feces of pre-weaned dairy calves. The AMD considered were ceftiofur, administered parenterally, and neomycin sulfate added in milk replacer and fed to calves during the first 20 days of life. Fifty-five calves, aged one to three days, were enrolled and followed to 64 days. Fecal samples were collected three times/week and treatments recorded daily. Enterobacteriaceae were quantified for a subset of 33 calves using spiral plating on plain, ceftiofur supplemented, and neomycin supplemented MacConkey agar. Negative binomial models were used to predict the association between treatment with AMD and the gain and loss of Enterobacteriaceae resistance over time. Acquisition of resistance by the Enterobacteriaceae occurred during treatment and peaked between days three to four post-treatment before decreasing to below treatment levels at days seven to eight post-treatment. Acquisition of neomycin resistance was observed on the first sampling day (day four from the start of feeding medicated milk replacer) to day eight, followed by cyclical peaks until day 29, when the Enterobacteriaceae counts decreased below pre-treatment. Enterobacteriaceae resistance against both AMD increased after AMD administration and didn't return to pre-therapeutic status until seven or more days after therapy had been discontinued. The study findings provide valuable insights into the dynamics of Enterobacteriaceae under routine AMD use in calves.

Antibiotic Resistance Genes and Associated Phenotypes in Escherichia coli and Enterococcus from Cattle at Different Production Stages on a Dairy Farm in Central California.

The objectives of this study were to characterize overall genomic antibiotic resistance profiles of fecal Escherichia coli and Enterococcus spp. from dairy cattle at different production stages using whole-genome sequencing and to determine the association between antimicrobial resistance (AMR) phenotypes and their corresponding genotypes. The Comprehensive Antibiotic Resistance Database (CARD) and ResFinder, two publicly available databases of antimicrobial resistance genes, were used to annotate isolates. Based on the ResFinder database, 27.5% and 20.0% of tested E. coli isolates (n = 40) harbored single and ≥3 antimicrobial resistance genes, respectively; for Enterococcus spp., we observed 87.8% and 8.2%, respectively. The highest prevalence of AMR genes in E. coli was for resistance to tetracycline (27.5%), followed by sulphonamide (22.5%) and aminoglycoside (20.0%); the predominant antimicrobial resistance genes in Enterococcus spp. targeted macrolide drugs (77.6%). Based on the CARD database, resistance to ≥3 antimicrobial classes was observed in all E. coli and 77.6% in Enterococcus spp. isolates. A high degree of agreement existed between the resistance phenotype and the presence of resistance genes for various antimicrobial classes for E. coli but much less so for isolates of Enterococcus. Consistent with prior work, fecal E. coli and Enterococcus spp. isolates from calves harbored a wide spectrum of resistance genes, compared to those from cattle at other production stages, based on the cross-sectional samples from the studied farm.