Preliminary In Vitro Evaluation of Silver, Copper and Gold Nanoparticles as New Antimicrobials for Pathogens That Induce Bovine Locomotion Disorders.

Lameness is a crucial problem in dairy farming. It worsens the welfare of cattle, reduces the milk yield, and causes economic losses. The etiology of lameness is varied and the cattle's condition may be infectious or non-infectious. The aim of this research was to analyze the biocidal properties of silver (AgNPs), gold (AuNPs), and copper (CuNPs) nanoparticles against bacteria causing lameness in cattle. The isolated pathogens used were Aerococcus viridans, Corynebacterium freneyi, Corynebacterium xerosis, and Trueperella pyogenes. The tested concentrations of nanoparticles were 50, 25, 12.5, 6.25, 3.125, and 1.56 mg/L. The methods used included the isolation of pathogens using standard microbiological procedures and their identification using mass spectrometry, physicochemical analysis, transmission electron microscopy, and cytotoxicity tests. Studies have shown that AgNPs at 3.125 and 1.56 mg/L concentrations, and CuNPs at 25 and 12.5 mg/L concentrations, have strong biocidal properties, while AuNPs have the weakest antimicrobial properties. The very limited number of in vivo studies focusing on lameness prevention in cattle indicate that new solutions need to be developed. However, further studies are necessary to evaluate if nanoparticles (NPs) may, in the future, become components of innovative biocides used to prevent lameness in dairy cattle.

Effect of GnRH treatment as a potential solution for ovarian disorders in dairy cows infected with foot and mouth disease in Indonesian smallholder farms.

Background: The outbreak of foot and mouth disease (FMD) in Indonesia induces reproductive disorders in dairy cows that lead to economic losses to smallholder dairy farms. Aim: The study was to assess the influence of FMD on reproductive traits and evaluate the effect of gonadotropin hormone-releasing hormone (GnRH) administrations on the reproductive performance in FMD-infected dairy cows. Methods: The study was conducted in Jemowo village, Taman Sari sub-district, Boyolali district, Central Java, Indonesia. A total of 155 cows were used to identify the reproductive disorders on FMD-infected dairy cows aged 2-10 years old. Cows were raised in similar conditions and fed diets. A single dose of 2 ml GnRH was injected intramuscularly into 96 ovarian disorder cows. Reproductive performance was measured by service per conception (S/C), conception rate (CR), and pregnancy rate (PR). A descriptive study was conducted to demonstrate the results. Results: The study showed that 61.9% of FMD-infected cows had reproductive disorders, whereby 53.5% ovarian hypofunction, 4.52% silent heat, 1.94% repeat breeder, 1.29% ovarian atrophy, and 0.65% endometritis. FMD-infected cows injected with GnRH had a 98% reproductive recovery rate. Moreover, the S/C, CR, and PR of cows injected with GnRH were 2.02%, 51%, and 85%. Conclusion: GnRH administrations enhanced the reproductive traits of FMD-infected dairy cows indicated by the improvement of CR and PR.

Quantifying trade-offs between therapeutic efficacy and resistance dissemination for enrofloxacin dose regimens in cattle.

The use of antimicrobial drugs in food-producing animals contributes to the selection pressure on pathogenic and commensal bacteria to become resistant. This study aims to evaluate the existence of trade-offs between treatment effectiveness, cost, and the dynamics of resistance in gut commensal bacteria. We developed a within-host ordinary differential equation model to track the dynamics of antimicrobial drug concentrations and bacterial populations in the site of infection (lung) and the gut. The model was parameterized to represent enrofloxacin treatment for bovine respiratory disease (BRD) caused by Pastereulla multocida in cattle. Three approved enrofloxacin dosing regimens were compared for their effects on resistance on P. multocida and commensal E. coli: 12.5 mg/kg and 7.5 mg/kg as a single dose, and 5 mg/kg as three doses. Additionally, we explored non-FDA-approved regimes. Our results indicated that both 12.5 mg/kg and 7.5 mg/kg as a single dose scenario increased the most the treatment costs and prevalence of P. multocida resistance in the lungs, while 5 mg/kg as three doses increased resistance in commensal E. coli bacteria in the gut the most out of the approved scenarios. A proposed non-FDA-approved scenario (7.5 mg/kg, two doses 24 h apart) showed low economic costs, minimal P. multocida, and moderate effects on resistant E. coli. Overall, the scenarios that decrease P. multocida, including resistant P. multocida did not coincide with those that decrease resistant E. coli the most, suggesting a trade-off between both outcomes. The sensitivity analysis suggests that bacterial populations were the most sensitive to drug conversion factors into plasma ( ß ), elimination of the drug from the colon ( ϑ ), fifty percent sensitive bacteria (P. multocida) killing effect ( L s50 ), fifty percent of bacteria (E. coli) above ECOFF killing effect ( C r50 ), and net drug transfer rate in the lung ( γ ) parameters.

Diagnostic utility of acute phase proteins and their ability to guide antibiotic usage in pigs, horses, and cattle: a mapping review.

To mitigate the use of antibiotics for many of the multifactorial diseases seen in pigs, horses and cattle, new diagnostic tools are needed. Acute phase protein (APP) measurements can, in humans, be used to guide antibiotic treatment initiation, evaluate treatment efficacy, and make a prognosis. The aim of this review is to collect evidence on the clinical functionality of APP measurements as a tool to guide antibiotic treatment in pigs, horses, and cattle. Literature was retrieved using Medline, CAB Abstracts and Google Scholar. The acute phase response has been investigated for a plethora of diseases and clinical signs and the major acute phase proteins are elevated in diseased compared to healthy animals. Few studies correlated acute phase response with aetiology, antibiotic treatment efficacy, prognosis, or severity of disease. The existing research does not support that APP can be used to guide antibiotic treatment, but the reported studies indicate that C-reactive protein (CRP) might be able to differentiate between bacterial and non-bacterial causes of disease in pigs. Serum amyloid A (SAA) might reflect underlying aetiology in horses and infectious or non-infectious cases of mastitis in cows.

Effect of recombinant bovine granulocyte colony-stimulating factor treatment during the peripartum period on postpartum diseases, reproductive performance, and milk production in Holstein cattle.

To boost the immune function around parturition, recombinant bovine granulocyte colony-stimulating factor (rbG-CSF) has been used to increase the number of neutrophils. Therefore, the aim of this study was to quantify the effect of rbG-CSF administration on the incidence of postpartum pathologies, reproductive performance, and milk production during the first three months of lactation. A total of 199 Holstein cows from one herd were included and were randomly allocated into two groups: Control (n = 103) and rbG-CSF (n = 96). Cows in the rbG-CSF group received 2 doses of a rbG-CSF injectable formulation, one 7 days before the expected date of calving and the other within 24 h after calving. For 6 weeks following calving, animals were examined weekly to assess the presence of postpartum pathologies. Milk production, protein and fat content, and somatic cell count were determined monthly by the regional dairy herd improvement association. Data about the reproductive performance were collected from on-farm software. To analyse the effect of treatment on the incidence of postpartum pathologies, Pearson's χ2 test and multivariable logistic regressions were performed. The effect on reproductive performance was analysed using Cox proportional hazard regression analysis for days open, binary logistic regression for first service conception rate and Oneway ANOVA test for the number of artificial inseminations. The effects of treatment on milk yield and milk composition were checked using GLM repeated measures analysis. No statistically significant differences were observed between treatment groups for any of the parameters evaluated. Only parity had a significant effect on days open and milk production (p < 0.05). In conclusion, in the present study no evidence was found that rbG-CSF could have an effect on the reproductive and productive parameters evaluated.

Administering the maternal bovine appeasing substance improves overall productivity and health in high-risk cattle during a 60-d feedlot receiving period.

This experiment evaluated health, physiological, and performance responses of high-risk cattle administered the maternal bovine appeasing substance (mBAS) during feedlot receiving. Angus-influenced, newly weaned male calves (n = 120) were purchased from an auction facility. Calves arrived at the research feedyard on day -1 and body weight (BW) was recorded upon arrival (199 ±â€…1 kg). Calves were ranked by arrival BW and received 1 of 2 treatments prior to initial processing (day 0): (1) 10 mL of an mBAS (Ferappease; FERA Diagnostics and Biologicals; College Station, TX) or (2) 10 mL of mineral oil (CON; placebo). Treatments were applied topically to the nuchal skin area (5 mL) and above the muzzle (5 mL). Calves were vaccinated against Clostridium and respiratory pathogens, dewormed, implanted, band-castrated, and received metaphylaxis at initial processing, and then distributed into 10 drylot pens according to arrival BW and treatment (n = 12 calves/pen, 5 pens/treatment). Calves were reapplied treatments (mBAS or CON) concurrently with booster vaccination on d 14. Feed intake and incidence of bovine respiratory disease (BRD) were recorded daily. Blood and hair samples from the tail-switch were collected on days 0, 14, 28, 42, and 60 for analysis of physiological variables. Calves were sampled for nasal microbiota analysis via swab on days 0, 14, and 28. Final shrunk BW was recorded on day 61 after 16 h of feed and water restriction. Calf BW gain and final BW did not differ between treatments (P ≥ 0.40). Incidence of BRD was similar (P = 0.99) between mBAS and CON (56.7% for both treatments; SEM = 6.45). A greater (P = 0.04) proportion of mBAS calves diagnosed with BRD required a single antibiotic treatment to regain health (70.6 vs. 47.0%; SEM = 8.32), and mortality was greater (P = 0.03) in CON calves diagnosed with BRD (17.6 vs. 2.94%; SEM = 5.133). Relative abundance of Mycoplasma in nasal microbiota was reduced (P = 0.04) in mBAS calves compared with CON (34.7 vs. 27.4%; SEM = 2.35). Cortisol concentration in hair from the tail-switch was less (P = 0.01) on day 28 for mBAS calves compared with CON. Administering mBAS improved (P = 0.04) total pen-based liveweight change during the experiment (498 vs. 309 kg/pen; SEM = 65.2) and final pen-based total liveweight (2,676 vs. 2,484 kg/pen; SEM = 65.4). Administration of mBAS to high-risk cattle decreased physiological stress markers, reduced mortality, and increased pen-based productivity during a 60-d receiving period.

Beef cattle are exposed to several stressors during the initial days in the feedlot, which impair their immunity and lead to bovine respiratory disease (BRD). Strategies to mitigate stress upon feedlot arrival are warranted, including administration of the maternal bovine appeasing substance (mBAS). The mBAS is a mixture of fatty acids that replicate the composition of the original bovine appeasing pheromone. Administering mBAS has been shown to alleviate the physiological consequences elicited by stressful management procedures in beef cattle. In this study, male beef calves were purchased from a commercial auction facility soon after weaning, transported to the research feedyard, and assigned initial processing within a 48-h period. Calves were castrated and received a metaphylactic antibiotic treatment during initial processing, which are relevant management practices in US feedyards. In general, mBAS administration at initial processing and 14 d later decreased physiological stress markers, improved immunity parameters, and reduced mortality by 83% during a 60-d feedlot receiving period. Calf growth and BRD incidence were not affected, but the reduced mortality from mBAS administration increased total liveweight production by the end of the experiment. Therefore, this study provides additional evidence of the benefits from administering mBAS to beef cattle upon feedlot arrival.

Anthelmintic resistance of gastrointestinal nematodes in cattle in Brazil and Argentina - current status and global perspectives.

This review outlines the current state of anthelmintic resistance (AHR) of gastrointestinal nematodes (GINs) among cattle in Argentina and Brazil, emphasizing the economic repercussions, animal health and welfare. The analysis explores factors associated with AHR and proposes a potential solution: the use of drug combinations. Both countries are grappling with a severe AHR scenario in cattle, having progressed through incipient, established, and advanced phases, leading to extreme cases of animal mortality due to ineffective control strategies. Genera such as Cooperia and Haemonchus have the highest reports of resistance, with Oesophagostomum radiatum also posing significant problems. While oral benzimidazoles and levamisole remain effective in most herds, moxidectin is entering an advanced resistance phase, and avermectins are increasingly deemed ineffective. The review explores the impact ofclimate, mixed grazing, animal movement and other husbandry practices, and the relationship between ectoparasite control and the emergence of resistant helminths. Notably, the discussion includes the strategic use of drug combinations as a valuable approach to address resistant GINs control in livestock, highlighting its significant potential to mitigate the challenges posed by AHR in the cattle industry of these countries.

A registry-based comparative analysis of antibiotic usage reporting for adult cattle on Danish dairy farms.

BACKGROUND: Antimicrobial resistance (AMR) is a significant global health concern, necessitating the monitoring of antimicrobial usage (AMU). However, there is a lack of consensus on the standardized collection and reporting of AMU data in the veterinary field. In Denmark, the Danish Cattle Database (DCDB) contains treatment information on animal level, which allows counting of number of treatments carried out, used daily doses (UDD). The Danish VetStat database (VetStat) contains information on veterinary medicinal prescriptions at farm level and uses fixed standard doses of each product to calculate number of daily treatments, animal daily doses (ADD). This study aimed to compare two different numerators, UDD and ADD, used to describe AMU on Danish cattle farms, and estimate their correlation. RESULTS: Routinely collected registry data from conventional dairy farms in Denmark for 2019 were used, including a total of 2,197 conventional dairy farms. The data from VetStat and the DCDB were aggregated and analysed, and treatment frequencies (TF) were calculated for both UDD and ADD, adjusting for farm size. Spearman correlation analysis and Bland-Altman plots were employed to assess the relationship and agreement between TF for ADD and UDD, respectively. The results showed a high correlation between TF for ADD and UDD for most prescription groups, i.e., groups used to categorise antibiotics based on target organs. An exception is found for the Udder prescription group, where a systematic underreporting of UDD compared to ADD was observed. This discrepancy may be due to combination treatments, and potential missing or grouped registrations in the DCDB. CONCLUSIONS: Our UDD and ADD comparison yields valuable insights on farm-level AMU. We observe strong correlations between UDD and ADD, except for udder treatments, where some farms report only 1/3 UDD compared to ADD, indicating potential underreporting. Further investigations are needed to understand the factors contributing to these patterns and to ensure the accuracy and completeness of recorded information. Standardizing AMU data collection and reporting remains crucial to tackle the global challenge of AMR effectively.

Acaricide effect of plants from the Brazilian savanna on a population of Rhipicephalus microplus with phenotypic resistance to cypermethrin and trichlorfon.

Rhipicephalus microplus is among the most important ectoparasites for livestock. The use of synthetic acaricides has raised some concerns due to the selection of tick populations that are resistant to acaricides and environmental contamination. Therefore, plant extracts have been used as alternatives for the treatment of animals infested with ticks. In this study, R. microplus populations from seven different dairy farms were collected and assessed for their resistance to the acaricides cypermethrin or trichlorfon. Larvae of the most resistant population were used in assays to evaluate the acaricide effect of leaf extracts from plants of the Brazilian savanna. The most active extracts were also tested against fully engorged females. Among seven tick populations, five and three showed resistance level ≥ III for cypermethrin or trichlorfon, respectively. The most resistant tick population was evaluated in mortality assays with the plants Piptadenia viridiflora, Annona crassiflora, Caryocar brasiliense, Ximenia americana, and Schinopsis brasilienses. The ethanolic extracts of C. brasiliense, X. americana and S. brasilienses showed higher larvicidal effects in comparison to the other extracts and cypermethrin. The ethanolic extract of X. americana showed 60.79 % efficacy against fully engorged females of the acaricide resistant tick strain. The ethanolic extracts of C. brasiliense, X. americana, and S. brasilienses showed peaks in HPLC-DAD, indicating the presence of tannins and flavonoids. Three of the plants showed promising results and should be explored in further studies to develop novel tools to control R. microplus in cattle.

Histological changes of oocytes of the cattle tick Rhipicephalus microplus (Canestrini, 1888) treated with copper solutions.

Infections caused by the ectoparasite Rhipicephalus microplus can cause major health problems in cattle, including death. Tick control is regularly made using a range of acaricide products. As a consequence, tick populations have been heavily selected for drug resistance. The objective of this work was to determine the in vitro efficacy of copper chloride and sulfate (CuCl2 and CuSO4) solutions against R. microplus. The adult immersion test (AIT), which measures the egg-laying and egg-hatch effects, was used for the Cu-II solutions at 30, 60, 120, 240, 480, and 1000 mM, in triplicates. Distilled water and the combination of cypermethrin 20% and chlorpyrifos 50% were used as controls. Histological sections were performed from the ovaries of adult engorged female ticks treated with 240, 480, and 1000 mM of CuCl2 and CuSO4. We have established a histological index of the damage caused by the solutions to the tick oocytes. The overall efficacy (egg laying & egg hatch) for CuCl2 and CuSO4 was 81.3, 82.5, 89.8, 84.5, 100.0, and 100%, and 61.7, 43.4, 62.5, 93.1, 100.0, and 98.5% respectively. Smaller oocytes were found in the Cu-II groups compared to the negative control. The histological data showed a concentration-dependent degenerative lesion of oocytes, described as cytoplasmic vacuolation and nuclear disorganization. The combination of cypermethrin and chlorpyriphos showed 100% efficacy. Cu-II solutions showed in vitro efficacy against adult engorged ticks being particularly harmful to oocytes. Thus, bioactive metals could be a complementary biofriendly treatment to control R. microplus and these injuries could be responsible for preventing egg hatch, and reducing pasture contamination. Safety studies are underway demonstrating the Cu-II potential in naturally infected cattle and their persistence in the environment.

Lentinan Ameliorates ß-Hydroxybutyrate-Induced Lipid Metabolism Disorder in Bovine Hepatocytes by Upregulating the Expression of Acetyl-coenzyme A Acetyltransferase 2.

Ketosis in dairy cows is often accompanied by the dysregulation of lipid homeostasis in the liver. Acetyl-coenzyme A acetyltransferase 2 (ACAT2) is specifically expressed in the liver and is important for regulating lipid homeostasis in ketotic cows. Lentinan (LNT) has a wide range of pharmacological activities, and this study investigates the protective effects of LNT on ß-hydroxybutyrate (BHBA)-induced lipid metabolism disorder in bovine hepatocytes (BHECs) and elucidates the underlying mechanisms. BHECs were first pretreated with LNT to investigate the effect of LNT on BHBA-induced lipid metabolism disorder in BHECs. ACAT2 was then silenced or overexpressed to investigate whether this mediated the protective action of LNT against BHBA-induced lipid metabolism disorder in BHECs. Finally, BHECs were treated with LNT after silencing ACAT2 to investigate the interaction between LNT and ACAT2. LNT pretreatment effectively enhanced the synthesis and absorption of cholesterol, inhibited the synthesis of triglycerides, increased the expression of ACAT2, and elevated the contents of very low-density lipoprotein and low-density lipoprotein cholesterol, thereby ameliorating BHBA-induced lipid metabolism disorder in BHECs. The overexpression of ACAT2 achieved a comparable effect to LNT pretreatment, whereas the silencing of ACAT2 aggravated the effect of BHBA on inducing disorder in lipid metabolism in BHECs. Moreover, the protective effect of LNT against lipid metabolism disorder in BHBA-induced BHECs was abrogated upon silencing of ACAT2. Thus, LNT, as a natural protective agent, can enhance the regulatory capacity of BHECs in maintaining lipid homeostasis by upregulating ACAT2 expression, thereby ameliorating the BHBA-induced lipid metabolism disorder.

Early stage Fasciola hepatica infection in growing cattle: impact on weight gain and liver weight determined in flukicide efficacy studies.

Although the negative impact of liver fluke (Fasciola hepatica) infection on production and health in cattle is generally accepted, results of individual research have been variable, ranging from important negative impacts on the animal to minimal or no impact. To add information on the impact of F. hepatica infection in growing cattle, weight gain and liver weight of young experimentally infected animals from seven controlled efficacy studies were analyzed. In each study, fluke naïve animals were inoculated with approximately 450 to 500 F. hepatica encysted metacercariae, blocked on body weight and randomly assigned into one untreated group (controls) and groups which were administered an experimental flukicide when the flukes were 4 weeks old (migrating) and sacrificed 8 weeks thereafter (12 weeks after inoculation). Data of groups which demonstrated >90% reduction of fluke counts following treatment and groups left untreated (total 103 and 47 animals, respectively) were compared. There was a significant (p < 0.0001) negative association between fluke count and weight gain while fluke count and liver weight and fluke count and relative liver weight were positively associated (p < 0.0001). Over the 8-week post-treatment period, flukicide-treated cattle had almost 15% more weight gain than the controls (50.9 kg vs. 44.4 kg; p = 0.0003). Absolute and relative liver weight was significantly (p < 0.0001) lower in flukicide-treated compared to untreated cattle. Overall, this analysis provided evidence of a substantial negative effect of early (migrating) liver fluke infection on the growth of young cattle, likely due to pathology of the liver and associated reduction in its function as the central organ for bioenergy and protein metabolism.

Anomaly detection in the veterinary antibiotic prescription surveillance system (IS ABV).

Antibiotic resistance is one of the major concerns in veterinary and human medicine and poses a considerable threat to both human and animal health. It has been shown that over- or misuse of antibiotics is one of the primary drivers of antibiotic resistance. To develop the surveillance of antibiotic use, Switzerland introduced the "Informationssystem Antibiotika in der Veterinärmedizin" (IS ABV) in 2019, mandating electronic registration of antibiotic prescriptions by all veterinarians in Switzerland. However, initial data analysis revealed a considerable amount of implausible data entries, potentially compromising data quality and reliability. These anomalies may be caused by input errors, inaccuracies, incorrect or aberrant master data or data transmission and make analysis impossible. To address this issue efficiently, we propose a two-stage anomaly detection framework utilizing machine learning algorithms. In this study, our primary focus was on cattle treatments with either single or group therapy, as they were the species with the highest prescription volume. However, not all outliers are necessarily incorrect; some may be legitimate but unusual antibiotic treatments. Thus, expert review plays a crucial role in distinguishing outliers, that are correct from actual errors. Initially, relevant prescription variables were extracted and pre-processed with a custom-built scaler. A set of unsupervised algorithms calculated the probability of each data point and identified the most likely outliers. In collaboration with experts, we annotated anomalies and established anomaly thresholds for each production type and active substance. These expert-annotated labels were then used to fine-tune the final supervised classification algorithms. With this methodology, we identified 22,816 anomalies from a total of 1,994,170 prescriptions in cattle (1.1 %). Cattle with no further specified production type had the most (2 %) anomalies with 7758 out of 379,995. The anomalies were consistently identified and comprised prescriptions with too high and too low dosages. Random Forest achieved a ROC-AUC score of 0.994, (95 % CI: 0.992, 0.995) and a F1-Score of 0.962 (95 % CI: 0.958, 0.966) for single treatments. The versatility of this framework allows its adaptation to other species within IS ABV and potentially to other prescription-based surveillance systems. If applied regularly to uploaded prescriptions, it should reduce input errors over time, improving the validity of the data in the long term.

An exploration of descriptive machine learning approaches for antimicrobial resistance: Multidrug resistance patterns in Salmonella enterica.

Salmonellosis is one of the most common foodborne diseases worldwide, with the ability to infect humans and animals. Antimicrobial resistance (AMR) and, particularly, multidrug resistance (MDR) among Salmonella enterica poses a risk to human health. Antimicrobial use (AMU) regulations in livestock have been implemented to reduce AMR and MDR in foodborne pathogens. In this study, we used an integrated machine learning approach to investigate Salmonella AMR and MDR patterns before and after the implementation of AMU restrictions in agriculture in the United States. For this purpose, Salmonella isolates from cattle in the National Antimicrobial Resistance Monitoring System (NARMS) dataset were analysed using three descriptive models consisting of hierarchical clustering, network analysis, and association rule mining. The analysis showed the impact of the United States' 2012 extra-label cephalosporin regulations on AMR trends and revealed a distinctive MDR pattern in the Dublin serotype. The results also indicated that each descriptive model provides insights on a specific aspect of resistance patterns and, therefore, combining these approaches make it possible to gain a deeper understanding of AMR.

Identification of genetic markers of resistance to macrolide class antibiotics in Mannheimia haemolytica isolates from a Saskatchewan feedlot.

Mannheimia haemolytica is a major contributor to bovine respiratory disease (BRD), which causes substantial economic losses to the beef industry, and there is an urgent need for rapid and accurate diagnostic tests to provide evidence for treatment decisions and support antimicrobial stewardship. Diagnostic sequencing can provide information about antimicrobial resistance genes in M. haemolytica more rapidly than conventional diagnostics. Realizing the full potential of diagnostic sequencing requires a comprehensive understanding of the genetic markers of antimicrobial resistance. We identified genetic markers of resistance in M. haemolytica to macrolide class antibiotics commonly used for control of BRD. Genome sequences were determined for 99 M. haemolytica isolates with six different susceptibility phenotypes collected over 2 years from a feedlot in Saskatchewan, Canada. Known macrolide resistance genes estT, msr(E), and mph(E) were identified in most resistant isolates within predicted integrative and conjugative elements (ICEs). ICE sequences lacking antibiotic resistance genes were detected in 10 of 47 susceptible isolates. No resistance-associated polymorphisms were detected in ribosomal RNA genes, although previously unreported mutations in the L22 and L23 ribosomal proteins were identified in 12 and 27 resistant isolates, respectively. Pangenome analysis led to the identification of 79 genes associated with resistance to gamithromycin, of which 95% (75 of 79) had no functional annotation. Most of the observed phenotypic resistance was explained by previously identified antibiotic resistance genes, although resistance to the macrolides gamithromycin and tulathromycin was not explained in 39 of 47 isolates, demonstrating the need for continued surveillance for novel determinants of macrolide resistance.IMPORTANCEBovine respiratory disease is the costliest disease of beef cattle in North America and the most common reason for injectable antibiotic use in beef cattle. Metagenomic sequencing offers the potential to make economically significant reductions in turnaround time for diagnostic information for evidence-based selection of antibiotics for use in the feedlot. The success of diagnostic sequencing depends on a comprehensive catalog of antimicrobial resistance genes and other genome features associated with reduced susceptibility. We analyzed the genome sequences of isolates of Mannheimia haemolytica, a major bovine respiratory disease pathogen, and identified both previously known and novel genes associated with reduced susceptibility to macrolide class antimicrobials. These findings reinforce the need for ongoing surveillance for markers of antimicrobial resistance to support improved diagnostics and antimicrobial stewardship.

Randomized clinical trial of ketoprofen or ceftiofur for treatment of metritis in dairy cows.

Our objectives were to compare the efficacy of ketoprofen or ceftiofur for treatment of metritis in dairy cows considering subsequent health, production, and reproduction. Cows from 2 commercial dairy farms in Ontario, Canada were examined with a Metricheck device 3 times per week from 2 to 14 DIM. Cows with metritis (fetid vaginal discharge; n = 193) were blocked by parity and fever (rectal temperature ≥39.5°C or <39.5°C) and within each block per farm, randomly assigned to receive 3 mg/kg BW of ketoprofen (KET) or 2.2 mg/kg of ceftiofur hydrochloride (CEF), once a day for 3 d. Day of enrollment was considered study d 0. Rectal temperature and attitude were evaluated in cows with metritis on study d 0, 3, 4, 7, 10, and 13, and vaginal discharge was evaluated on study d 4, 7, 10, and 13. Body condition was scored at enrollment and at 35 DIM, and serum concentration of haptoglobin was measured at d 0, 2, 4, and 7. Cows with rectal temperature ≥39.5°C or a depressed attitude on d 3 were classified as clinical failure and received treatment with ceftiofur for 3 d (KET), or 2 additional days (CEF), to a maximum of 5 d of treatment with ceftiofur. At 35 ± 3 DIM cows were examined for uterine involution via transrectal palpation, purulent vaginal discharge (PVD) via Metricheck, and endometritis via endometrial cytology. Time to onset of cyclicity was assessed by serum progesterone (P4) measurements at 28, 42, and 56 DIM. Contemporary cows from the same farms without metritis (NOMET; n = 1,043) were used for comparison. Data were analyzed with mixed linear or logistic regression or Cox's proportional hazard models, including herd as a random effect. The proportion of clinical resolution of metritis on d 3 (96% vs. 92%), of cows with fever (from d 3 to d 13 after enrollment) or fetid discharge (from d 4 to d 13 after enrollment), and the number of medical treatments (3.1 vs. 3.3) were not different between CEF and KET, respectively. Cows in KET received fewer antibiotic treatments than cows in CEF (0.3 vs. 3.1). Uterine involution, prevalence of PVD (50% vs. 47%) and subclinical endometritis (6.6% vs. 4.3%), and proportion of cyclic cows (82% vs. 86%) did not differ between CEF and KET. Cows in KET had greater serum haptoglobin concentration from d 2 to 7 after enrollment. The incidence of mastitis, lameness, or displaced abomasum to 60 DIM and subclinical ketosis to 21 DIM did not differ among CEF, KET, and NOMET. There were no differences in median time to first AI (CEF = 68 d, 95% CI: 65-70; KET = 69 d, 95% CI: 68-72; NOMET = 69 d, 95% CI: 68-70), median time to pregnancy (CEF = 118 d, 95% CI: 92-145; KET = 113 d, 95% CI: 90-135; NOMET = 105 d, 95% CI: 101-109), pregnancy at first AI at 33 d after insemination (CEF = 42%; KET = 41%; NOMET = 41%), pregnancy loss after first AI (CEF = 8%; KET = 11%; NOMET = 8%), hazard of pregnancy, or hazard of culling up to 300 DIM. Milk yield was not different between CEF and KET during the first 10 wk, but was lesser in KET at wk 2 and 4 and CEF at wk 2, 4, and 6 than in NOMET. In this pilot-scale study, given early detection, we did not detect differences in subsequent health, milk yield, or reproductive performance in cows with metritis initially treated for 3 d with CEF or KET. Additional, larger studies are warranted.

Virulent-MDR-ESBL E. coli and Klebsiella pneumoniae report from North Sinai calves diarrhea and in vitro antimicrobial by Moringa oleifera.

The health of calves has a significant impact on the production of cows and livestock. Some desert plants have pharmacological importance, as they can be used to reduce antibiotic resistance. Our hypothesis is designed to detect Virulent- Multidrug-Resistant and Extended- spectrum Beta- lactamase Enterobacteriaceae (Virulent-MDR-ESBL Enterobacteriaceae and to determine whether Moringa oleifera has antibacterial activity against the detected isolates. A total of 39 Enterobacteriaceae isolates from 28 diarrheic samples were collected from calves aged between 20 days and 20 months from 3 different flocks in North Sinai, Sahl-Eltina region, Egypt. E.coli 46% (18/39), O157 13% (5/39), Klebsiella pneumoniae 41% (16/39). MDR members accounted for 87%, while ESBL isolates accounted for 43%. The antibacterial activity is represented by microdilution. Minimum inhibition concentration (MIC) for the methanol extract of Moringa oleifera ranged from 2.5,5,10, and 25mg/ ml among E.coli isolates, and O157 was susceptible to (2.5mg/ ml), Klebsiella pneumoniae isolates were susceptible to (5-50mg/ ml). Analysis of the methanol extract revealed that ferulic acid was the dominant phenolic compound with a concentration of 29,832 parts per million (ppm). In silico docking study expected the active site of ferulic acid to act on the tyrosine bacterial enzyme through Pi-alkyl, Pi-anion, Carbon hydrogen bonds, and extra ionic attractive interactions with copper ions which can stabilize ferulic acid inside the targeted pocket Diverse virulent gene profiles were observed in E. coli. The Shiga toxin-producing Escherichia coli (STEC) was reported in 83% of the isolated E. coli, while the DNA gyrase (gyrA) was harbored in 100% of Klebsiella pneumoniae isolates. Various profiles of antibiotic resistance genes for both E. coli and Klebsiella pneumoniae isolates were distinguished. blaTEM genes were detected in 99% of E. coli and 100% of Klebsiella pneumoniae. Sequence analysis for E. coli strain DRC-North Sinai-Eg was placed in accession numbers (OP955786) for the Shiga toxin 2 gene (Stx2A), (OP997748) and (OP997749) for the Adhesion to host cell gene (Eae). For the hemolysine gene (hylA), the accession number was (OP946183). Klebsiella pneumoniae strain DRC-North Sinai-Eg was placed in (OP946180) for (gyrA). This study has proven the broad range of Moringa oliefera's antibacterial effects in vitro against the virulent-MDR- ESBL E. coli and Klebsiella pneumoniae isolated from North Sinai calves diarrhea. These are congruent with the disability effect on bacterial tyrosinase enzyme through docking study therefore, we recommend the usage of this desert plant as a prospective feed additive, we endorse this as an antibacterial new insight natural source and for the medication of considered pathogens with zoonotic impacts.

Development potential of selenium in the prevention and treatment of bovine endometritis.

Endometritis is a common postpartum disease in cows. It delays uterine involution and impairs normal physiological function. This can result in long-term or even lifelong infertility and cause significant losses to the dairy farming industry. Traditional treatments like antibiotics possess certain shortcomings, such as antibiotic residues, the abuse of antibiotics, and increased antimicrobial resistance of pathogens. Alternative treatment strategies are needed to minimize the utilization of antibiotics in dairy production. As an essential trace element in animals, selenium (Se) plays a vital role in regulating immune function, the inflammatory response, and oxidative stress, affecting the speed and completeness of tissue repair. This paper reviewed previous studies to analyse the potential of Se in the prevention and treatment of bovine endometritis, aiming to provide a new direction to increase production capacity in the future.

Knowledge, attitudes, practices and risk perception of cattle farmers in Namibia to antibiotic use and resistance.

The misuse of antibiotics in livestock contributes to the emergence of antibiotic resistant bacteria. The development of antibiotic resistance in bacteria of animal origin is of public health concern since drug resistant pathogens in animals can spread to humans. Several countries have therefore introduced regulations to restrict the use of antibiotics in livestock. Namibia was the first African country to ban the use of antibiotics as growth promoters and restrict the use of antibiotics to the treatment of bacterial diseases with a prescription from a veterinarian. However, there are limited studies on antibiotic stewardship among farmers in the country. The aim of this study was to determine the antibiotic awareness and usage of cattle farmers in Oshikoto region Namibia. A questionnaire on antibiotic knowledge, attitudes and practices (KAP) was administered to 274 randomly selected cattle farmers in Namibia. To calculate the KAP scores, one mark was awarded for each correct answer and zero was awarded for incorrect or unsure responses. Respondents having ≥ 50% correct answers were considered as having good KAP scores. The KAP score ranged from 8 to 73%. Forty-three per cent of the respondents had good KAP scores. Good KAP was associated with age, farm type and education status. Semi-commercial/commercial farmers had significantly better KAP than post and homestead farmers. The findings of this study indicate that there is a need to improve antibiotic usage and resistance education programmes in order to promote rational antibiotic usage.

Inefficacy of ivermectin and moxidectin treatments against Dictyocaulus viviparus in dairy calves.

BACKGROUND: The bovine lungworm Dictyocaulus viviparus negatively impacts bovine health and leads to substantial economic losses. Lungworm infections can be difficult to manage due to the unpredictable and severe nature of clinical outbreaks. Despite the widespread use of macrocyclic lactones (MLs) in grazing cattle in the UK, there have been no confirmed reports of resistant lungworms to date, with only one case of anthelmintic-resistant (ML) lungworm confirmed worldwide. METHODS: Lungworm Baermann filtrations were conducted on first-season grazing dairy calves as part of a wider study investigating anthelmintic resistance in gastrointestinal nematodes in Scotland using the faecal egg count reduction test. RESULTS: Clinical signs and significant numbers of lungworm larvae in faeces were observed after treatment with either ivermectin or moxidectin. LIMITATIONS: There are no established guidelines for the diagnosis of resistant lungworms in the field. Currently, resistance can only be diagnosed after a controlled efficacy test has been conducted. This limits the conclusions that can be drawn; however, they are highly suggestive of resistance. CONCLUSION: This short report describes the inefficacy of ivermectin and moxidectin against D. viviparus and is highly suggestive of ML resistance.