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Antimicrobial resistance (AMR) is one of the greatest threats to global health worldwide and is threatening not only humans, but also animal production systems, including dairy farms. The objective of this paper was to describe risks factors associated with AMR on dairy farms in Québec, Canada. This observational cross-sectional study included 101 commercial dairy farms and took place between the springs of 2017 and 2018 for a one-year period. We explored risk factors such as farm practices and producer's knowledge (measured using a questionnaire), antimicrobial use (quantified using veterinary invoices), and the presence of Salmonella Dublin (tested by serology). We evaluated AMR with fecal Escherichia coli retrieved from pre-weaned calves and lactating cows using the following outcomes: the presence of extended-spectrum-ß-lactamase/AmpC resistance and the number of resistances to antimicrobial classes. We used logistic regression models to evaluate the association between each risk factor and the 2 outcomes for the 2 types of samples (pre-weaned calves and lactating cows). Furthermore, we explored the relationships between these risk factors utilizing data dimensionality reduction and hierarchical clustering. Outputs of these analyses were used as regressors for AMR in regression models. While the results for univariate analyses were ambiguous, the unsupervised analysis naturally categorized the sample of farms according to their health/treatment status (dimension 1, explaining 12.9% of the variance) and herd size (dimension 2, explaining 7.8%). Three clusters of farms were identified (cluster 1: mainly healthy herds and low ceftiofur users, cluster 2: relatively high ceftiofur users, cluster 3: farms with a higher incidence of diseases and higher antimicrobial treatment rates). Dimension 1 and cluster membership were statistically associated with the presence of extended-spectrum-ß-lactamase/AmpC resistance in lactating cows and in pre-weaned calves. Dimension 1 was also statistically associated with the number of resistances in lactating cows and in pre-weaned calves. This study highlights the complexity of analyzing risk factors associated with AMR. Our results suggest that the herd health status and the AMU-related practices used are associated with AMR in dairy farms. However, prospective studies are needed to confirm a causal relation.
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The globalization in plant material trading has caused the emergence of invasive pests in many ecosystems, such as the alder pathogen Phytophthora ×alni in European riparian forests. Due to the ecological importance of alder to the functioning of rivers and the increasing incidence of P. ×alni-induced alder decline, effective and accessible decision tools are required to help managers and stakeholders control the disease. This study proposes a Bayesian belief network methodology to integrate diverse information on the factors affecting the survival and infection ability of P. ×alni in riparian habitats to help predict and manage disease incidence. The resulting Alder Decline Network (ADnet) management tool integrates information about alder decline from scientific literature, expert knowledge and empirical data. Expert knowledge was gathered through elicitation techniques that included 19 experts from 12 institutions and 8 countries. An original dataset was created covering 1189 European locations, from which P. ×alni occurrence was modeled based on bioclimatic variables. ADnet uncertainty was evaluated through its sensitivity to changes in states and three scenario analyses. The ADnet tool indicated that mild temperatures and high precipitation are key factors favoring pathogen survival. Flood timing, water velocity, and soil type have the strongest influence on disease incidence. ADnet can support ecosystem management decisions and knowledge transfer to address P. ×alni-induced alder decline at local or regional levels across Europe. Management actions such as avoiding the planting of potentially infected trees or removing man-made structures that increase the flooding period in disease-affected sites could decrease the incidence of alder disease in riparian forests and limit its spread. The coverage of the ADnet tool can be expanded by updating data on the pathogen's occurrence, particularly from its distributional limits. Research on the role of genetic variability in alder susceptibility and pathogen virulence may also help improve future ADnet versions.
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Alnus , Teorema de Bayes , Doenças das Plantas/microbiologia , Doenças das Plantas/estatística & dados numéricos , Phytophthora , Ecossistema , Europa (Continente)/epidemiologia , Florestas , Conservação dos Recursos NaturaisRESUMO
The continuous improvement of expression platforms is necessary to respond to the increasing demand for recombinant proteins that are required to carry out structural or functional studies as well as for their characterization as biotherapeutics. While transient gene expression (TGE) in mammalian cells constitutes a rapid and well-established approach, non-clonal stably transfected cells, or "pools," represent another option, which is especially attractive when recurring productions of the same protein are required. From a culture volume of just a few liters, stable pools can provide hundreds of milligrams to gram quantities of high-quality secreted recombinant proteins.In this chapter, we describe a highly efficient and cost-effective procedure for the generation of Chinese Hamster Ovary cell stable pools expressing secreted recombinant proteins using commercially available serum-free media and polyethylenimine (PEI) as the transfection reagent. As a specific example of how this protocol can be applied, the production and downstream purification of recombinant His-tagged trimeric SARS-CoV-2 spike protein ectodomain (SmT1) are described.
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Cricetulus , Polietilenoimina , Proteínas Recombinantes , Glicoproteína da Espícula de Coronavírus , Transfecção , Células CHO , Animais , Proteínas Recombinantes/genética , Proteínas Recombinantes/biossíntese , Proteínas Recombinantes/isolamento & purificação , Proteínas Recombinantes/metabolismo , Transfecção/métodos , Polietilenoimina/química , Glicoproteína da Espícula de Coronavírus/genética , Glicoproteína da Espícula de Coronavírus/metabolismo , Glicoproteína da Espícula de Coronavírus/biossíntese , Glicoproteína da Espícula de Coronavírus/isolamento & purificação , SARS-CoV-2/genética , SARS-CoV-2/metabolismo , Cricetinae , Meios de Cultura Livres de SoroRESUMO
Mastitis is one of the most common diseases of dairy cattle. It has a high impact on farm economy, farmers' working time, and antimicrobial usage (AMU). Selective dry cow therapy (SDCT) is an effective means of reducing AMU without negatively affecting udder health. The objective of our study was to evaluate the impact of SDCT implementation on farmer's income, working time, and AMU, using a bioeconomic model. A stochastic dairy simulation model (DairyHealthSim) based on a weekly model was used to simulate herd dynamics, reproduction, milk production, culling decisions, health outcomes, and the management of health events. A specific module was developed for the simulation of quarter-level intramammary infection (IMI) acquisition and elimination during the lactation and dry-off periods, and 25 different farm settings were defined to represent herds with various udder health situations. We then defined 20 scenarios of SDCT by combining both the use of different thresholds of somatic cell count and milk bacteriology for treatment allocation and the use of internal teat sealant (ITS). All SDCT protocols had a low impact on farmer's income, and we identified some protocols with a positive farm gross margin (up to 15.83 CA$/dried- cow). We also found that adding an ITS to all cows led to greater economic gain. The application of SDCT had a low impact on farmers' working time, except when milk bacteriology was used for decision-making. Antimicrobial treatment to all cows above 200,000 cells/mL at last control, with the use of ITS on all cows, seems a good choice in most dairy farms. These findings could be used to convince farmers to adopt this strategy at dry-off.
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Despite good manufacturing practices and rigorous cleaning and sanitizing procedures established in dairy processing plants, microbiological contamination remains the main cause of products being non-compliant and/or atypical and hence not fit for human consumption. The objective of this study was to isolate, identify and characterize bacteria, yeasts and molds associated with substandard dairy products in Canada and to create a collection of reference isolates. In addition to conventional microbiological characterization, each isolate was tested for biofilm-forming ability and susceptibility to heat, antimicrobial agents, and common industrial disinfectants. Among the 105 microbial strains isolated from pasteurized milk, cream, and cheese samples, 24 bacterial isolates, belonging mainly to the genus Pseudomonas, were shown to be moderate or strong biofilm producers in 96-well plates and highly resistant to peracetic acid (100 ppm, 5 min contact time) and sodium hypochlorite (70 ppm, 5 min contact time). In addition, 56 bacterial isolates, including Acinetobacter baumannii, Enterobacter bugandensis, Klebsiella pneumoniae and Pseudomonas spp., were found resistant to ampicillin, fosfomycin and/or ceftriaxone, while 14 others, such as Bacillus spp. and Macrococcus spp., withstood a heat treatment equivalent to low-temperature long-time pasteurization (63°C for 30 min). This descriptive study provides new information on potential problematic microorganisms in dairies and will guide the development of novel control strategies intended to prevent and reduce microbiological contamination and the associated economic losses.
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The diagnosis of infectious diseases at herd level can be challenging as different stakeholders can have conflicting priorities. The current study proposes a "proof of concept" of an approach that considers a reasonable number of criteria to rank plausible diagnostic strategies using multi-criteria decision analysis (MCDA) methods. The example of Salmonella Dublin diagnostic in Québec dairy herds is presented according to two epidemiological contexts: (i) in herds with no history of S. Dublin infection and absence of clinical signs, (ii) in herds with a previous history of infection, but absence of clinical signs at the moment of testing. Multiple multiparty exchanges were conducted to determine: 1) stakeholders' groups; 2) the decision problem; 3) solutions to the problem (options) or diagnostic strategies to be ordered; 4) criteria and indicators; 5) criteria weights; 6) the construction of a performance matrix for each option; 7) the multi-criteria analyses using the visual preference ranking organization method for enrichment of evaluations approach; 8) the sensitivity analyses, and 9) the final decision. A total of nine people from four Québec's organizations (the dairy producers provincial association along with the DHI company, the ministry of agriculture, the association of veterinary practitioners, and experts in epidemiology) composed the MCDA team. The decision problem was "What is the optimal diagnostic strategy for establishing the status of a dairy herd for S. Dublin infection when there are no clinical signs of infection?". Fourteen diagnostic strategies composed of the three following parameters were considered: 1) biological samples (bulk tank milk or blood from 10 heifers aged over three months); 2) sampling frequencies (one to three samples collection visits); 3) case definitions to conclude to a positive status using imperfect milk- or blood-ELISA tests. The top-ranking diagnostic strategy was the same in the two contexts: testing the bulk tank milk and the blood samples, all samples collected during one visit and the herd being assigned a S. Dublin positive status if one sample is ELISA-positive. The final decision favored the top-ranking option for both contexts. This MCDA approach and its application to S. Dublin infection in dairy herds allowed a consensual, rational, and transparent ranking of feasible diagnostic strategies while taking into account the diagnostic tests accuracy, socio-economic, logistic, and perception considerations of the key actors in the dairy industry. This promising tool can be applied to other infectious diseases that lack a well-established diagnostic procedure to define a herd status.
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Doenças dos Bovinos , Indústria de Laticínios , Técnicas de Apoio para a Decisão , Salmonelose Animal , Animais , Bovinos , Salmonelose Animal/diagnóstico , Salmonelose Animal/epidemiologia , Quebeque/epidemiologia , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/microbiologia , Feminino , Salmonella enterica/isolamento & purificaçãoRESUMO
Klebsiella pneumoniae species complex members, particularly K. pnemoniae sensu stricto, are common bovine clinical mastitis pathogens and often the cause of hospital- and community-acquired infections in humans. Here, we present 148 draft genome assemblies and annotations of K. pneumoniae species complex members from bovine and human hosts in Canada.
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The global food animal industry faces a growing concern regarding antimicrobial resistance (AMR), primarily driven by the use of antimicrobials (AMs) for the treatment, control, and prevention of diseases. Addressing this challenge requires promoting responsible antimicrobial use (AMU) practices. In 2019, the province of Québec, Canada, took a significant step by implementing a regulation that limits the use of AMs of very high importance for human medicine (category I AMs as defined by Health Canada) in the food animal industry. However, the implementation of such regulation can significantly influence behavioral shifts among producers, contributing to the wider effort against AMR. Therefore, the objective of this observational study was to describe the perceived changes in knowledge of dairy producers and on-farm practices following the implementation of this regulation, using a cohort design. Data collection involved administering questionnaires to 87 dairy producers from 3 regions of the province of Québec (Estrie, Montérégie, Centre-Du-Québec) before (2017-2018) and after (2020-2021) the implementation of the regulation. The questionnaires explored the descriptive characteristics of farms, the knowledge of producers about the categorization of AMs, their on-farm treatment practices, and the perceived impacts of the regulation. Statistical analysis included t-tests and McNemar tests to compare the paired data obtained using the 2 questionnaires. The results indicated an increase in the knowledge score (the number of AMs correctly categorized by the producers by their importance for human medicine) after the implementation of the regulation, suggesting an improved understanding of the categorization of AMs based on their importance for human medicine. Trends in AMU practices for treating clinical mastitis and reproductive diseases suggested that category I AMs were less likely to be reported as the primary treatment after the regulation, while category II AMs were more often reported as primary treatment. Adoption of the selective dry cow therapy method significantly increased, while the use of teat sealants remained unchanged. Moreover, producers had divergent perceptions regarding the effect of the regulation on the cure rates and disease frequencies. This disparity emphasizes the need for comprehensive data collection to discern the risks associated with such regulatory shifts. The study acknowledges several limitations, including the potential for recall bias, confirmation bias, and desirability bias. Despite these limitations, this study shows that implementing regulations to encourage responsible AMU drives positive transformations in producers' knowledge and on-farm practices. This underscores the pivotal impact of proactive interventions in combating the escalating threat of AMR within the global food animal industry.
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This study aimed to estimate the prevalence of poultry aspergillosis and evaluate the accuracy of histopathology (test under evaluation) and mycological culture (an imperfect reference test). Farms raising layer and breeder or broiler birds, with suspected aspergillosis cases, clinical or subclinical, were eligible and visited for sampling. After necropsy, histopathology and mycological culture examinations were conducted by two evaluators. A Bayesian latent class model was used to estimate the accuracy of histopathology when compared to the imperfect reference test, mycological culture. A total of 142 chicken farms, 96 laying and breeding hen farms, and 46 broiler farms were used for the study. True aspergillosis median prevalence was estimated at 63.7% (95% credibility intervals, CrI: 53.8%, 73.0%) in layers and breeders and at 65.2% (95% CrI: 50.2%, 78.3%) in the broiler farms' population. The median diagnostic sensitivity of histopathology and culture were estimated at, respectively, 98.8% (95% CrI: 94.6%, 100.0%) and 90.4% (95% CrI: 83.6%, 95.3%). Tests' diagnostic specificity was estimated at, respectively, 97.3% (95% CrI: 87.7%, 99.9%) and 95.7% (95% CrI: 91.8%, 98.2%). Both tests had very high and comparable positive predictive values, but, in a population where disease prevalence was 25%, histopathology had a higher negative predictive value than culture.
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Our objective was to assess potassium monopersulfate as a disinfectant used in footbath to control digital dermatitis (DD) in dairy cows. We hypothesized that a potassium monopersulfate solution would control DD. A 180-day randomized negative controlled trial was conducted in a 265-Holstein free-stall facility. Throughout the trial, foot bathing was performed bi-weekly using a split (left vs. right feet) footbath: one tub filled with 1% potassium monopersulfate (treatment), the other with tap water (control). Digital dermatitis lesions were scored during trimming chute examinations of the unwashed hind heels every 90 days using the modified M-scoring system. Digital dermatitis lesions were re-categorized into four variables: 1) inactive; 2) active; 3) any; 4) inactive or absence of DD lesions. Three longitudinal outcomes were characterized: risks of 1) developing a DD lesion; 2) reactivating an inactive DD lesion; 3) development of an inactive or the absence of the DD lesion. A generalized linear model was used to compare the variables and longitudinal outcomes between treated and control groups. Prevalence of active DD lesions increased from 12.5% to 39.9% between days 0 and 90. This significant increase in prevalence justified the discontinuation of the study on day 90 for ethical reasons. There was no statistical difference between treated and control groups for the first outcome (RR: 1.0; 95% CI: 0.62, 1.7), the second outcome (RR: 1.0; 95% CI: 0.62,-1.7); or the third outcome (RR: 0.88; 95% CI: 0.37, 2.1). A 1% potassium monopersulfate footbath solution appears ineffective to control DD in this study.
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Doenças dos Bovinos , Dermatite Digital , Doenças do Pé , Casco e Garras , Compostos de Potássio , Sulfatos , Feminino , Bovinos , Animais , Lactação , Dermatite Digital/tratamento farmacológico , Dermatite Digital/prevenção & controle , Dermatite Digital/epidemiologia , Doenças dos Bovinos/tratamento farmacológico , Doenças dos Bovinos/prevenção & controle , Doenças dos Bovinos/epidemiologia , Casco e Garras/patologia , Indústria de Laticínios , Doenças do Pé/prevenção & controle , Doenças do Pé/veterináriaRESUMO
Morbidity and mortality remain important concerns for veal production which traditionally involves very young calves being transported directly from dairy farms to calf-raising facilities or sold through auction markets from dairy farms. There are few studies that examine the relationship between transfer of passive immunity and health outcomes in veal production in Québec. The objective of this study was to investigate the risk factors associated with morbidity and mortality in milk and grain-fed veal facilities in Québec. Between October 2017 and December 2018, a prospective cohort study was conducted on 59 batches of milk- and grain-fed veal calves from Montérégie and Centre du Québec regions in Québec, Canada. A total of 30 calves per batch were randomly sampled for the transfer of passive immunity (TPI) status using the Brix refractometer (serum threshold ≥ 8.4%). Throughout the production cycle, arrival, treatment, mortality, and slaughter records of each batch of calves were extracted and used to quantify individual- and group-level risk factors. Morbidity and mortality were modelled through generalized linear mixed models (GLMMs) using a logit link, as function of categorical variables (individual inadequate TPI, arrival season, purchasing sites, and arrival weights) and a continuous variable (proportion of inadequate TPI in the batch). Given that the health and performance data are usually collected retrospectively under commercial settings, incomplete data was recognized as a potential issue. Thus, multiple imputation models were used. From 59 batches where 1729 calves were sampled for measuring TPI, 1084 calves had a serum Brix% < 8.4% giving a prevalence of 62.7% of inadequate TPI. The prevalence of morbidity and mortality in those 59 batches were not associated with the batch-level inadequate TPI prevalence. On the other hand, individual calves with inadequate TPI had higher odds of morbidity (OR: 1.56; 95% CI: 1.2-2.05, P = 0.001) compared to those calves with adequate TPI. Odds of mortality, on the other hand, were not significantly affected by inadequate TPI (OR:1.27; 95% CI: 0.74-2.18, P = 0.3). Calves arriving to the facility during the winter season had lower odds of mortality than those arriving in fall (OR:0.16; 95% CI: 0.03-0.78, P = 0.02). In conclusion, this study suggests that inadequate TPI, as assessed upon arrival in veal farms, remains an important concern for negative health outcomes in veal calves.
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Doenças dos Bovinos , Carne Vermelha , Animais , Bovinos , Estudos Prospectivos , Quebeque/epidemiologia , Estudos Retrospectivos , Canadá , Doenças dos Bovinos/epidemiologiaRESUMO
Although recycled manure solids (RMS) bedding is used on dairy farms, it could allow bacterial growth when contaminated by feces and thus increase the incidence of clinical mastitis in cows. The objective of this study was to describe bacterial growth in three different types of RMS bedding, as well as in sand, when samples were experimentally inoculated with Escherichia coli and Klebsiella pneumoniae. Two 3-day trials were conducted, during which treatments included inoculating bedding samples with E. coli and K. pneumoniae, as well as no inoculation. The trial was repeated 3 times for each bedding sample on each day. Samples were incubated at 15°C for 3 d and bacterial counts were measured every day. After inoculation, there was no significant K. pneumoniae or E. coli growth phase during the trial in those RMS samples that were prepared either in a container or in a heap. Recycled manure solids and sand samples prepared in a rotary drum, however, showed a similar active growth phase of K. pneumoniae during the first 24 h of the trial. Moreover, a significant E. coli growth phase was observed in the samples of sand bedding in the first 24 h. The 3 different types of RMS bedding samples did not react in a similar manner to coliform inoculation. No active growth phase was observed in bedding samples already containing a high bacterial concentration following inoculation with coliforms.
Bien que la litière de fumier recyclé (LFR) soit utilisée dans les fermes laitières, elle pourrait permettre la croissance bactérienne lorsqu'elle est contaminée par des matières fécales et augmenter ainsi l'incidence de mammite clinique chez les vaches. L'objectif de cette étude était de décrire la croissance bactérienne dans trois types de LFR, ainsi que dans du sable, lorsque des échantillons étaient inoculés expérimentalement avec Escherichia coli et Klebsiella pneumoniae. Deux essais de trois jours ont été réalisés, au cours desquels les échantillons de litière ont été inoculés ou non avec E. coli et K. pneumoniae. L'essai a été répété trois fois pour chaque échantillon de litière, chaque jour. Les échantillons ont été incubés à 15 °C pendant 3 jours et la numération bactérienne a été mesurée chaque jour. Après inoculation, il n'y a pas eu de phase de croissance significative de K. pneumoniae ou d'E. coli au cours de l'essai dans les échantillons de LFR préparés dans un conteneur ou en tas. Les échantillons de sable et de LFR préparés dans un tambour rotatif ont cependant montré une phase de croissance active similaire de K. pneumoniae pendant les premières 24 heures de l'essai. En outre, une phase de croissance significative d'E. coli a été observée dans les échantillons de litière de sable au cours des premières 24 h. Les trois différents types d'échantillons de LFR n'ont pas réagi de la même manière à l'inoculation de coliformes. Aucune phase de croissance active n'a été observée dans les échantillons de litière contenant déjà une concentration bactérienne élevée après l'inoculation de coliformes.(Traduit par Docteur Simon Dufour).
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Doenças dos Bovinos , Mastite Bovina , Feminino , Bovinos , Animais , Klebsiella pneumoniae , Esterco/microbiologia , Escherichia coli , Areia , Abrigo para Animais , Roupas de Cama, Mesa e Banho/veterinária , Mastite Bovina/microbiologiaRESUMO
As clinical mastitis (CM) treatments are responsible for a large portion of antimicrobial use on dairy farms, many selective CM treatment protocols have been developed and evaluated against a blanket treatment approach of CM cases. Selective treatment protocols use outcomes of diagnostic tests to exclude CM cases from antimicrobial treatment when they are unlikely to benefit. To tailor interventions to increase uptake of selective treatment strategies, a comprehension of current on-farm treatment practices and factors affecting treatment decisions is vital. Two questionnaires were conducted among 142 farms across 5 provinces participating in the Canadian Dairy Network for Antimicrobial Stewardship and Resistance in this cross-sectional study. Self-reported adoption of selective CM treatments by dairy farmers was 64%, with median of 82% of cows treated in those herds using selective treatment. Using logistic regression models, the odds to implement a selective CM treatment protocol increased with a decreasing average cow somatic cell count. No other associations were identified between use of a selective CM treatment protocol and farm characteristics (herd size, CM incidence, province, milking system, and housing system). Three subsets of farmers making cow-level CM treatment decisions were identified using a cluster analysis approach: those who based decisions almost exclusively on severity of clinical signs, those who used various udder health indicators, and farmers who also incorporated more general cow information such as production, age, and genetics. When somatic cell count was considered, the median threshold used for treating was >300,000 cells/mL at the last Dairy Herd Improvement test. Various thresholds were present among those considering CM case history. Veterinary laboratories were most frequently used for bacteriological testing. Test results were used to start, change, and stop treatments. Regardless of protocol, reasons for antimicrobial treatment withheld included cow being on a cull list, having a chronic intramammary infection, or being at end of lactation (i.e., close to dry off). If clinical signs persisted after treatment, farmers indicated that they would ask veterinarians for advice, stop treatment, or continue with the same or different antibiotics. Results of this study can be used to design interventions targeting judicious mastitis-related antimicrobial use, and aid discussions between veterinarians and dairy producers regarding CM-related antimicrobial use.
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Anti-Infecciosos , Doenças dos Bovinos , Mastite Bovina , Animais , Bovinos , Feminino , Humanos , Antibacterianos/uso terapêutico , Anti-Infecciosos/uso terapêutico , Canadá , Doenças dos Bovinos/tratamento farmacológico , Contagem de Células/veterinária , Estudos Transversais , Indústria de Laticínios/métodos , Fazendas , Glândulas Mamárias Animais , Mastite Bovina/tratamento farmacológico , Mastite Bovina/epidemiologia , LeiteRESUMO
Salmonellosis is one of the leading causes of gastrointestinal infections in humans. In Canada, it is estimated that approximately 87,500 cases of salmonellosis occur every year in humans, resulting in 17 deaths. In the United States, it is estimated that 26,500 hospitalizations and 420 deaths occur every year. In dairy cattle, infections caused by nontyphoidal Salmonella enterica can cause mild to severe disease, including enteritis, pneumonia, and septicemia. Our study objectives were to determine the proportion of fecal samples positive for Salmonella in dairy cattle in Canada and determine the resistance pattern of these isolates. We used data collected through the Canadian Dairy Network for Antimicrobial Stewardship and Resistance (CaDNetASR). Pooled fecal samples from preweaning calves, postweaning heifers, lactating cows, and manure storage were cultured for Salmonella, and the isolates were identified using matrix-assisted laser desorption/ionization-time of flight mass spectrometry. Antimicrobial susceptibilities were determined using the minimum inhibitory concentration test, and resistance interpretation was made according to the Clinical and Laboratory Standards Institute. A 2-level, multivariable logistic regression model was built to determine the probability of recovering Salmonella from a sample, accounting for province, year, and sample source. The proportion of farms with at least one positive sample were 12% (17/140), 19% (28/144), and 17% (24/144) for the sampling years 2019, 2020, and 2021, respectively. Out of the 113 Salmonella isolates, 23 different serovars were identified. The occurrence of Salmonella appeared to be clustered by farms and provinces. The most common serovars identified were Infantis (14%) and Typhimurium (14%). Overall, 21% (24/113) of the Salmonella isolates were resistant to at least one antimicrobial. Resistance to tetracycline was commonly observed (17%); however, very limited resistance to category I antimicrobials (categorization according to Health Canada that includes third-generation cephalosporins, fluoroquinolones, polymyxins, and carbapenems) was observed, with one isolate resistant to amoxicillin and clavulanic acid. The proportion of Salmonella isolates resistant to 2 and 3 antimicrobial classes was 3.5% and 8.8%, respectively. Our study provided valuable information on the proportion of fecal samples positive for Salmonella, the serovars identified, and the associated resistance patterns across CaDNetASR herds, at regional and national levels.
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Anti-Infecciosos , Salmonelose Animal , Salmonella enterica , Humanos , Bovinos , Animais , Feminino , Antibacterianos/farmacologia , Antibacterianos/uso terapêutico , Farmacorresistência Bacteriana , Lactação , Canadá , Salmonelose Animal/epidemiologia , Indústria de Laticínios/métodos , Fezes , Salmonella , Anti-Infecciosos/farmacologia , Testes de Sensibilidade Microbiana/veterinária , Farmacorresistência Bacteriana MúltiplaRESUMO
SARS-CoV-2 subunit vaccines continue to be the focus of intense clinical development worldwide. Protein antigens in these vaccines most commonly consist of the spike ectodomain fused to a heterologous trimerization sequence, designed to mimic the compact, prefusion conformation of the spike on the virus surface. Since 2020, we have produced dozens of such constructs in CHO cells, consisting of spike variants with different mutations fused to different trimerization sequences. This set of constructs displayed notable conformational heterogeneity, with two distinct trimer species consistently detected by analytical size exclusion chromatography. A recent report showed that spike ectodomain fusion constructs can adopt an alternative trimer conformation consisting of loosely associated ectodomain protomers. Here, we applied multiple biophysical and immunological techniques to demonstrate that this alternative conformation is formed to a significant extent by several SARS-CoV-2 variant spike proteins. We have also examined the influence of temperature and pH, which can induce inter-conversion of the two forms. The substantial structural differences between these trimer types may impact their performance as vaccine antigens.
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COVID-19 , SARS-CoV-2 , Animais , Cricetinae , Humanos , Vacinas contra COVID-19/genética , Temperatura , Cricetulus , Antígenos , Mutação , Concentração de Íons de Hidrogênio , Anticorpos NeutralizantesRESUMO
There is currently no perfect test for determining herd-level status for Salmonella Dublin in dairy cattle herds. Our objectives were to evaluate the accuracy, predictive ability, and misclassification cost term of different testing scenarios using repeated measurements for establishing the S. Dublin herd status. Diagnostic strategies investigated used repeated bulk tank milk antibody-ELISA tests, repeated rounds of blood antibody-ELISA tests on non-lactating animals or a combination of both approaches. Two populations hypothesized to have different S. Dublin prevalences were included: (i) a convenience sample of 302 herds with unknown history of infection; and (ii) a cohort of 58 herds that previously tested positive to S. Dublin. Bulk milk samples were collected monthly for 6-7 months and serum were obtained from 10 young animals on two occasions, at the beginning and end of bulk milk sampling period. A series of Bayesian latent class models for two populations and comparing two tests were used to compare bulk milk-based to serum-based strategies. Moreover, Monte Carlo simulations were used to compared diagnostic strategies combining both types of samples. For each diagnostic strategy, we estimated the predictive values using two theoretical prevalences (0.05 and 0.25). Misclassification cost term was also estimated for each strategy using these two prevalences and a few relevant false-negative to false-positive cost ratios. When used for screening a population with an expected low prevalence of disease, for instance for screening herds with no clinical signs and no previous S. Dublin history, a diagnostic strategy consisting of two visits at 6 months interval, and with herd considered positive if bulk milk PP% ≥ 35 and/or ≥ 1/10 animals are positive on one or both visits could be used to confidently rule-out S. Dublin infection (median negative predictive value of 0.99; 95% Bayesian credible intervals, 95BCI: 0.98, 1.0). With this approach, however, positive results should later be confirmed with more specific tests to confirm whether S. Dublin is truly present (median positive predictive value of 0.36; 95BCI: 0.22, 0.57). The same diagnostic strategy could also be used confidently to reassess the S. Dublin status in herds with a previous S. Dublin history. When use for such a purpose, the predictive value of a positive result could be greatly improved, from 0.78 (95BCI: 0.65, 0.90) to 0.99 (95BCI: 0.94, 1.0) by requiring ≥ 1 positive result on both visits, rather than at any of the two visits.
Assuntos
Doenças dos Bovinos , Salmonelose Animal , Humanos , Bovinos , Animais , Leite/química , Teorema de Bayes , Anticorpos Antibacterianos/análise , Doenças dos Bovinos/diagnóstico , Doenças dos Bovinos/epidemiologia , Salmonelose Animal/diagnóstico , Salmonelose Animal/epidemiologia , Salmonella , Ensaio de Imunoadsorção Enzimática/veterinária , Ensaio de Imunoadsorção Enzimática/métodos , ImunoglobulinasRESUMO
Antimicrobial resistance in pathogenic bacteria is one of the preeminent concerns for the future of global health. There is a dose-dependent relationship between antimicrobial use (AMU) and the prevalence of antimicrobial-resistant pathogens. As most AMU in Canada is related to animal agriculture, there is a need to reduce overall AMU, which could be accomplished through surveillance of AMU in animal agriculture, including the dairy industry. The objective of this study was to quantify AMU on dairy farms across Canada. This study had two parts: a description of data collected in 2019-2020, and a meta-analysis comparing this data to previous estimates of AMU in the Canadian dairy industry. The first included a garbage can audit (GCA) on 107 farms in four Canadian provinces (British Columbia, Alberta, Ontario, and Nova Scotia) in 2020; AMU data were converted to the dose-based metrics of defined course doses (DCD) and defined daily doses (DDD). Mixed-effect linear models were fit to determine the relationship between province and use of different classes of antimicrobials. On average, for every 100 animals on the farm, 117 DCD of antimicrobials were administered per year (IQR: 55, 158). These treatments amounted to 623 DDD / 100 animal-yr (IQR: 302, 677 DDD/100 animal-years). Penicillins were the most used class of antimicrobials, followed by first-and third-generation cephalosporins. Farms in Ontario used more third-generation cephalosporins than other provinces. The second part of this study compared AMU in 2020 to previously reported Canadian studies through a meta-analysis. A GCA was conducted in 2007-2008 in Alberta, Ontario, Québec, and the Maritime provinces (Prince Edward Island, New Brunswick and Nova Scotia); another GCA was conducted in Québec in 2018. Overall, AMU was lower in 2018-2020 than in 2007-2008, with the exception of third-generation cephalosporin use, which increased.
RESUMO
Infections with antimicrobial resistant pathogens are a major threat to human and animal health worldwide. Further, reduction of livestock-associated antimicrobial use (AMU) is often identified as an area of focus. Selective dry cow therapy (DCT) warrants consideration as an important way to decrease AMU on Canadian dairy farms. In addition, teat sealants (TS) are a nonantimicrobial alternative for prevention of intramammary infection during the dry period. Therefore, objectives of this study were to determine how antimicrobials and TS are used at dry-off on Canadian dairy farms to determine selective DCT uptake and enacted selection protocols. It was expected that these data will provide a baseline understanding of DCT practices and highlight areas for future intervention to further reduce AMU. An observational study was conducted utilizing 2 in-person questionnaires conducted between July 2019 and September 2021 on 144 participating dairy farms in 5 Canadian provinces (British Columbia = 30, Alberta = 30, Ontario = 31, Québec = 29, and Nova Scotia = 24). Overall, 45 farms (31%) reported adopting selective DCT, 95 (66%) enacted blanket DCT, and 4 (3%) did not provide antimicrobial DCT. Farms enacting selective DCT had approximately 50% less intramammary antimicrobials used at dry-off compared with blanket DCT farms. Cow somatic cell count history was the most common criterion for selective DCT decision-making, followed by previous clinical mastitis history, bacteriological culture, and milk production. A slight majority of farms (56%) applied TS to all cows at dry-off, whereas 17 farms (12%) used TS selectively, and 46 farms (32%) did not use TS. Larger herds more often used TS, and farms with an automatic milking system more often used TS selectively than applied to all cows. Results highlighted the variability in antimicrobial treatment and TS use protocols at dry-off on Canadian dairy farms, and the potential for further antimicrobial reduction with increased adoption of selective DCT.
Assuntos
Anti-Infecciosos , Mastite Bovina , Feminino , Animais , Bovinos , Humanos , Fazendas , Seleção de Pacientes , Mastite Bovina/prevenção & controle , Mastite Bovina/tratamento farmacológico , Anti-Infecciosos/uso terapêutico , Leite , Ontário , Glândulas Mamárias Animais , Contagem de Células/veterinária , Antibacterianos/uso terapêutico , Lactação , Indústria de Laticínios/métodosRESUMO
Antimicrobial resistance (AMR) in animals, including dairy cattle, is a significant concern for animal and public health worldwide. In this study, we used data collected through the Canadian Dairy Network for Antimicrobial Stewardship and Resistance (CaDNetASR) to: (1) describe the proportions of AMR in fecal E. coli, and (2) investigate the relationship between antimicrobial use (AMU) (intramammary and systemic routes, while accounting for confounding by other variables) and AMR/multidrug resistance (MDR - resistance to ≥ 3 antimicrobial classes) in fecal E. coli from Canadian dairy farms. We hypothesized that an increase of the AMU was associated with an increase in AMR in E. coli isolates. A total of 140 dairy farms across five provinces in Canada were included in the study. Fecal samples from pre-weaned calves, post-weaned heifers, lactating cows, and farm manure storage were cultured, and E. coli isolates were identified using MALDI-TOF MS. The minimum inhibitory concentrations (MIC) to 14 antimicrobials were evaluated using a microbroth dilution methodology. AMU was quantified in Defined Course Dose (DCD - the dose for a standardized complete treatment course on a standard size animal) and converted to a rate indicator - DCD/100 animal-years. Of 1134 fecal samples collected, the proportion of samples positive for E. coli in 2019 and 2020 was 97.1% (544/560) and 94.4% (542/574), respectively. Overall, 24.5% (266/1086) of the E. coli isolates were resistant to at least one antimicrobial. Resistance towards tetracycline was commonly observed (20.7%), whereas resistance to third-generation cephalosporins, fluoroquinolones, and carbapenems was found in 2.2%, 1.4%, and 0.1% of E. coli isolates, respectively. E. coli isolates resistant to two or ≥ 3 antimicrobial classes (MDR) was 2.7% and 15%, respectively. Two multilevel models were built to explore risk factors associated with AMR with AMU being the main exposure. Systemic AMU was associated with increased E. coli resistance. For an increase in systemic AMU equivalent to its IQR, the odds of resistance to any antimicrobial in the model increased by 18%. Fecal samples from calves had higher odds of being resistant to any antimicrobial when compared to other production ages and farm manure storage. The samples collected in 2020 were less likely to be resistant when compared to samples collected in 2019. Compared to previous studies in dairy cattle in North America, AMR in E. coli was lower.