Biological and genomic characterization of 4 novel bacteriophages isolated from sewage or the environment using non-aureus Staphylococci strains.

Non-aureus staphylococci (NAS) are an essential group of bacteria causing antimicrobial resistant intramammary infections in livestock, particularly dairy cows. Therefore, bacteriophages emerge as a potent bactericidal agent for NAS mastitis. This study aimed to obtain NAS-specific bacteriophages using bacterial strains isolated from cows with mastitis, subsequently evaluating their morphological, genomic, and lytic characteristics. Four distinct NAS bacteriophages were recovered from sewage or the environment of Chinese dairy farms; PT1-1, PT94, and PT1-9 were isolated using Staphylococcus chromogenes and PT1-4 using Staphylococcus gallinarum. Both PT1-1 (24/54, 44 %) and PT94 (28/54, 52 %) had broader lysis than PT1-4 (3/54, 6 %) and PT1-9 (10/54, 19 %), but PT1-4 and PT1-9 achieved cross-species lysis. All bacteriophages had a short latency period and good environmental tolerance, including surviving at pH=4-10 and at 30-60℃. Except for PT1-9, all bacteriophages had excellent bactericidal efficacy within 5 h of co-culture with host bacteria in vitro at various multiplicity of infection (MOIs). Based on whole genome sequencing, average nucleotide identity (ANI) analysis of PT1-1 and PT94 can be classified as the same species, consistent with whole-genome synteny analysis. Although motifs shared by the 4 bacteriophages differed little from those of other bacteriophages, a phylogenetic tree based on functional proteins indicated their novelty. Moreover, based on whole genome comparisons, we inferred that cross-species lysis of bacteriophage may be related to the presence of "phage tail fiber." In conclusion 4 novel NAS bacteriophages were isolated; they had good biological properties and unique genomes, with potential for NAS mastitis therapy.

Hailey-Hailey disease: clinical, diagnostic and therapeutic update.

Hailey-Hailey disease is a rare genodermatosis described in 1939, with an autosomal dominant inheritance pattern, characterized by compromised adhesion between epidermal keratinocytes. It has an estimated prevalence of 1/50,000, with no gender or race predilection. It results from a heterozygous mutation in the ATP2C1 gene, which encodes the transmembrane protein hSPA1C, present in all tissues, with preferential expression in keratinocytes. Mutations in the ATP2C1 gene cause changes in the synthesis of junctional proteins, leading to acantholysis. It usually begins in adulthood, with isolated cases at the extremes of life. It manifests as vesico-bullous lesions mainly in the flexural areas, which develop into erosions and crusts. Chronic lesions may form vegetative or verrucous plaques. Pruritus, a burning feeling and pain are common. It evolves with periods of remission and exacerbation, generally triggered by humidity, friction, heat, trauma and secondary infections. The diagnosis is based on clinical and histopathological criteria: marked suprabasal acantholysis, loosely joined keratinocytes, giving the appearance of a "dilapidated brick wall", with a few dyskeratotic cells. The acantholysis affects the epidermis and spares the adnexal epithelia, which helps in the differential diagnosis with pemphigus vulgaris. Direct immunofluorescence is negative. The main differential diagnoses are Darier disease, pemphigus vegetans, intertrigo, contact dermatitis, and inverse psoriasis. There is no cure and the treatment is challenging, including measures to control heat, sweat and friction, topical medications (corticosteroids, calcineurin inhibitors, antibiotics), systemic medications (antibiotics, corticosteroids, immunosuppressants, retinoids and immunobiologicals) and procedures such as botulinum toxin, laser and surgery. There is a lack of controlled clinical trials to support the choice of the best treatment.

Extended-Spectrum Beta-Lactamase-Producing Escherichia coli and Other Antimicrobial-Resistant Gram-Negative Pathogens Isolated from Bovine Mastitis: A One Health Perspective.

Antimicrobial resistance (AMR) poses an imminent threat to global public health, driven in part by the widespread use of antimicrobials in both humans and animals. Within the dairy cattle industry, Gram-negative coliforms such as Escherichia coli and Klebsiella pneumoniae stand out as major causative agents of clinical mastitis. These same bacterial species are frequently associated with severe infections in humans, including bloodstream and urinary tract infections, and contribute significantly to the alarming surge in antimicrobial-resistant bacterial infections worldwide. Additionally, mastitis-causing coliforms often carry AMR genes akin to those found in hospital-acquired strains, notably the extended-spectrum beta-lactamase genes. This raises concerns regarding the potential transmission of resistant bacteria and AMR from mastitis cases in dairy cattle to humans. In this narrative review, we explore the distinctive characteristics of antimicrobial-resistant E. coli and Klebsiella spp. strains implicated in clinical mastitis and human infections. We focus on the molecular mechanisms underlying AMR in these bacterial populations and critically evaluate the potential for interspecies transmission. Despite some degree of similarity observed in sequence types and mobile genetic elements between strains found in humans and cows, the existing literature does not provide conclusive evidence to assert that coliforms responsible for mastitis in cows pose a direct threat to human health. Finally, we also scrutinize the existing literature, identifying gaps and limitations, and propose avenues for future research to address these pressing challenges comprehensively.

Prevalence and spatial distribution of infectious diseases of dairy cattle in Ontario, Canada.

We investigated the prevalence and spatial distribution of selected pathogens associated with infectious diseases of dairy cattle in Ontario, Canada. The cross-sectional study surveyed bulk tank milk for antibodies against bovine leukemia virus (BLV), Mycobacterium avium ssp. paratuberculosis (MAP), and Salmonella Dublin, and for the presence of mastitis pathogens (Staphylococcus aureus, Streptococcus agalactiae, Mycoplasma bovis). Between October 2021 and June 2022, bulk tank milk samples were obtained from every commercial dairy farm in Ontario (n = 3,286). Samples underwent ELISA testing for the presence of BLV, MAP, and S. Dublin antibodies, and quantitative PCR testing for the detection of specific antigens of pathogens associated with mastitis. Bayesian models were used to estimate prevalence, and spatial analysis was carried out to identify regional clusters of high pathogen prevalence. Prevalence varied for different pathogens, and BLV was widespread across dairy farms in Ontario, with an estimated prevalence of 88.3%. The prevalence of MAP, Staph. aureus and S. Dublin in Ontario dairy herds was 39.8%, 31.5%, and 5.1%, respectively. The vast majority of dairy herds in Ontario were free of intramammary infections caused by Strep. agalactiae and M. bovis. Clusters of increased positive test rates were detected for S. Dublin, MAP, and Staph. aureus, indicating potential geographic risk factors for pathogen transmission. For S. Dublin, an area of increased test positivity rates was detected in southwestern Ontario, close to the Canada-United States border where most of the dairy herds in Ontario are located. Conversely, a localized cluster of positive test outcomes involving 14 subdivisions located in the southeastern region of Ontario was detected for Staph. aureus. Findings from our survey highlight the importance of the testing of aggregated samples and conducting spatial analysis as part of disease surveillance programs, and for implementing risk-based trading approaches among dairy producers.

Accuracy of an AI-based automated plate reading mobile application for the identification of clinical mastitis-causing pathogens in chromogenic culture media.

Using on-farm microbiological culture (OFC), based on chromogenic culture media, enables the identification of mastitis causing pathogens in about 24 h, allows rapid decision making on selective treatment and control management measures of cows with clinical mastitis (CM). However, accurate interpretation of OFC results requires trained and experienced operators, which could be a limitation for the use of OFC in dairy farms. Our hypothesis was that AI-based automated plate reading mobile application can analyze images of microorganisms' colonies in chromogenic culture media with similar diagnostic performance as a trained specialist evaluator. Therefore, the aim of the present study was to evaluate the diagnostic accuracy of an AI-based application (Rumi; OnFarm, Piracicaba, São Paulo, Brazil) for interpreting images of mastitis causing microorganism colonies grown in chromogenic culture media. For this study two trials were organized to compare the results obtained using an AI-based application Rumi with the interpretation of: (1) a trained specialist, using MALDI-TOF MS as the gold standard; (2) farm personnel users (FPU). In trial 1, a total of 476 CM milk samples, from 11 farms located in São Paulo (n = 7) and Minas Gerais (n = 4), southeast Brazil, were inoculated in chromogenic culture media plates (Smartcolor 2, OnFarm, Piracicaba, São Paulo, Brazil) by specialists under lab conditions, and digital images were recorded 24 h after incubation at 37 °C. After that, all the 476 digital images were analyzed by the Rumi and by another specialist (who only had access to the digital images) and the diagnostic accuracy indicators sensitivity (Se) and specificity (Sp) were calculated using MALDI-TOF MS microbiological identification of the isolates as the reference. In Trial 2, a total of 208 CM milk samples, from 150 farms from Brazil, were inoculated in chromogenic culture media plates by FPU, and the results of microbiological growth were visually interpreted by FPU under on-farm conditions. After visual interpretation, results were recorded using an OnFarmApp application (herd manage application for mastitis by OnFarm, Piracicaba, São Paulo, Brazil), and the images of the chromogenic culture plates were captured by the OnFarmApp to be evaluated by Rumi and Bayesian Latent Class Models were performed to compare Rumi and the FPU. In Trial 1, Rumi presented high and intermediate accuracy results, with the only exception of the low Enterococcus spp.'s Se. In comparison with the specialist, Rumi performed similarly in Se and Sp for most groups of pathogens, with the only exception of non-aureus staphylococci where Se results were lower. Both Rumi and the specialist achieved Sp results > 0.96. In Trial 2, Rumi had similar results as the FPU in the Bayesian Latent Class Model analysis. In conclusion, the use of the AI-based automated plate reading mobile application can be an alternative for visual interpretation of OFC results, simplifying the procedures for selective treatment decisions for CM based on OFC.

Population-based genomic surveillance of Pseudomonas aeruginosa causing bloodstream infections in a large Canadian health region.

PURPOSE: Population-based surveillance was undertaken to determine clinical factors, susceptibility patterns, and incidence rates (IR) of Pseudomonas aeruginosa causing bloodstream infections (BSIs) in a Canadian region (2010-2018). METHODS: We combined clinical data with genomics to characterize P. aeruginosa (BSIs) (n = 167) in a well-defined Canadian (Calgary) human population over a 9-year period (2010-2018). RESULTS: The annual population IR per 100,000 patient years increased from 3.4/100,000 in 2010 to 5.9/100,000 in 2018, with the highest IRs in elderly males from the hospital setting. Over a quarter of patients presented with febrile neutropenia, followed by urinary tract infections and pneumonia. Antimicrobial resistance (AMR) rates and determinants were rare. The P. aeruginosa population was polyclonal consisting of three dominant sequence types (STs), namely ST244, ST111, and ST17. Antimicrobial-susceptible ST244 was the most common clone and belonged to three clades (A, B, C). The ST244 IR/100,000 increased over time due to the expansion of clade C. Multidrug-resistant ST111 was the second most common clone and IR/100,000 decreased over time. ST111 belonged to three clades (A, B, C) with clade C containing blaVIM-2. Different serotypes were linked to various STs. The IR/100,000 of P. aeruginosa that belonged to serotypes O6 increased significantly over time. CONCLUSION: An effective multivalent vaccine consisting of five serotypes (O1, O3, O5, O6, O11) would confer protection to > 70% of Calgary residents with P. aeruginosa BSIs. This study has provided a unique perspective of the population dynamics over time of P. aeruginosa STs, clades, and serotypes responsible for BSIs.

Comparative genomic analyses of Klebsiella pneumoniae K57 capsule serotypes isolated from bovine mastitis in China.

Klebsiella pneumoniae can cause severe clinical mastitis in dairy cows, with K. pneumoniae type K57 (K57-KP) being the most common capsular serotype. To identify virulence factors and antimicrobial-resistance (AMR) genes of K57-KP with varying virulence, Galleria mellonella (greater wax moth) larvae were infected as a screening model to characterize virulence of 90 K57-KP strains, with 10 and 11 strains defined as virulent or attenuated, respectively, based on larval survival rates. Next, virulence of these 21 isolates was subsequently confirmed in adhesion and lactate dehydrogenase release assays, using bovine mammary epithelial cells cultured in vitro. Finally, genes associated with virulence and AMR were characterize with whole-genome sequencing. These 21 K57-KP strains were designated into 16 sequence types based on multi-locus sequence typing and allocated in phylogenetic analysis based on single nucleotide polymorphisms. We found great genetic diversity among isolates. In addition, adhesion-associated genes (e.g., fimA, sfaA, and focA) aminoglycoside-resistance genes (aph(6)-Id, strAB) were associated with virulence. This study provided new knowledge regarding virulence of K57-KP associated with bovine mastitis, which may inform development of novel diagnostic tools and prevention strategies for bovine mastitis.

Relationship Between Celiac Disease and Dental Caries in Children and Adolescents: A Systematic Review and Meta-Analysis

ABSTRACT Objective: To assess the relationship between celiac disease and the prevalence of dental caries in children and adolescents and also evaluate if dental enamel defects in celiac children predispose to dental caries. Material and Methods: Searches were performed in the following databases: PubMed, Scopus, Cochrane Library, Latin American and Caribbean Literature on Health Sciences, and OpenGrey. The risk of bias was assessed using the Newcastle-Ottawa Scale. DMFT/dmft and DMFS/dmfs data of observational studies that compared the prevalence of caries between children and adolescents with celiac disease and healthy individuals. Meta-analysis was performed using a random effects model. Heterogeneity between studies was estimated using Cochran's Q test, and inconsistency was measured using I2 statistics. Results: Of the 121 studies retrieved, 17 were selected, and 12 were included in the meta-analysis. The prevalence of caries in the primary dentition (dmft) did not differ between celiac patients and controls [SMD = -0.35; 95% CI (-0.83; 0.13); p = 0,15; I2 = 89%]. There was also no difference in the prevalence of caries in permanent teeth (DMFT) between groups [SMD = -0.44; 95% CI (-1.02; 0.14); p = 0.14; I2 = 95%]. Conclusion: Celiac disease is not a determinant factor in the development of dental caries in children and adolescents compared to the control group.

Chapter 9.1: The Use of Fluorides in the Control of Coronal Caries.

Fluoride is the main agent used to control dental caries, with a very successful history after its widespread implementation worldwide, resulting in significant caries declines around the globe. In this chapter, the mechanism of action and peculiarities of different methods of fluoride use are revised. Fluoride controls caries progression by interfering with the processes of mineral exchanges (demineralization, remineralization) to which teeth are frequently subjected upon exposure to dental biofilm and a diet rich in fermentable sugars. Because caries is a ubiquitous disease of modern societies, different strategies have been developed to successfully keep fluoride available in oral fluids: (1) community-based approaches, such as water fluoridation, in which fluoride is delivered to communities at very low levels; (2) individual-based approaches, such as fluoride toothpastes, in which fluoride is delivered at higher concentrations, directly into the oral cavity, in a convenient combination with the act of brushing teeth to remove dental biofilm; (3) professionally-delivered approaches, such as fluoride gels and varnishes, in which very-high concentration products react with teeth resulting in local fluoride reservoirs that release the ion for weeks/months; and (4) combinations of these approaches, such as the concomitant use of fluoridated water and toothpaste, or the use of professional fluorides in caries-active patients. In conclusion, this chapter reviews the different modalities of fluoride use for coronal caries control, and summarizes the combined application of these methods according to the best evidence available.

A scoping review of the testing of bulk tank milk to detect nonbacterial pathogens or herd exposure to nonbacterial pathogens in dairy cattle.

In this scoping review, we characterized the literature reporting on the testing of bulk milk samples to detect microorganisms other than bacteria that can cause diseases in dairy cattle, including viruses, helminths, algae, and protozoa. A search strategy was completed by screening databases, conference proceedings, animal health agency websites, disease surveillance program websites, and handbooks of cattle-related diagnostic tests for potentially relevant articles. Two reviewers independently screened articles in English, Portuguese, or Spanish; original studies reporting on the testing of farm-level, unprocessed bulk milk samples for presence of pathogens or specific antibodies against agents other than bacteria that can cause diseases in cows were retained. From all studies, we used spreadsheets to extract relevant information, including pathogen screened, test used, and country of origin of bulk milk samples. Additionally, for studies reporting sufficient data to estimate test characteristics, we extracted detailed information about herd eligibility, testing protocol, and herd-level infection definition. A total of 8,829 records were identified, from which 1,592 were retained and assessed for eligibility, and 306 were included. Bovine viral diarrhea virus, Fasciola hepatica, Ostertagia ostertagi, and bovine herpesvirus 1 were the most frequently screened agents, reported from 107, 45, 45, and 33 studies, respectively. Sensitivity of bulk milk ELISA to detect herds with animals infected by bovine herpesvirus 1 ranged from 2 to 100%, and was affected mostly by antigen selection, cut-off adopted, herd vaccination status, and seroprevalence of lactating cows. Bulk milk ELISA had very high specificity to detect herds free of bovine leukemia virus, and varying sensitivity to detect herds with infected animals, which depended on the within-herd seroprevalence of lactating cattle. As for bovine viral diarrhea virus, in general, the sensitivity of bulk milk ELISA was moderate to high (>80%) when infection status was defined based on presence of persistently infected cattle or a high proportion of seropositive lactating cattle. Nevertheless, bulk milk ELISA was not able to distinguish infected and noninfected herds based on presence of seropositive unvaccinated weanlings. The PCR or quantitative PCR protocols employed had very low sensitivities (<40%) and very high specificities (>95%) to classify bovine viral diarrhea virus infection status of dairy herds. Sensitivity and specificity of bulk milk ELISA to classify herds with regards to presence of F. hepatica- or O. ostertagi-parasitized cattle were generally high and driven mostly by the definition of herd infection status. Conversely, bulk milk ELISA demonstrated varying characteristics to detect herds with or without Dictyocaulus viviparus-parasitized cattle, depending primarily on the antigen selected and presence of cattle with clinical signs of lungworm infection.

Invited review: Selective treatment of clinical mastitis in dairy cattle.

Treatment of clinical mastitis (CM) and use of antimicrobials for dry cow therapy are responsible for the majority of animal-defined daily doses of antimicrobial use (AMU) on dairy farms. However, advancements made in the last decade have enabled excluding nonsevere CM cases from antimicrobial treatment that have a high probability of cure without antimicrobials (no bacterial causes or gram-negative, excluding Klebsiella spp.) and cases with a low bacteriological cure rate (chronic cases). These advancements include availability of rapid diagnostic tests and improved udder health management practices, which reduced the incidence and infection pressure of contagious CM pathogens. This review informed an evidence-based protocol for selective CM treatment decisions based on a combination of rapid diagnostic test results, review of somatic cell count and CM records, and elucidated consequences in terms of udder health, AMU, and farm economics. Relatively fast identification of the causative agent is the most important factor in selective CM treatment protocols. Many reported studies did not indicate detrimental udder health consequences (e.g., reduced clinical or bacteriological cures, increased somatic cell count, increased culling rate, or increased recurrence of CM later in lactation) after initiating selective CM treatment protocols using on-farm testing. The magnitude of AMU reduction following a selective CM treatment protocol implementation depended on the causal pathogen distribution and protocol characteristics. Uptake of selective treatment of nonsevere CM cases differs across regions and is dependent on management systems and adoption of udder health programs. No economic losses or animal welfare issues are expected when adopting a selective versus blanket CM treatment protocol. Therefore, selective CM treatment of nonsevere cases can be a practical tool to aid AMU reduction on dairy farms.

Molecular Epidemiology of Global Carbapenemase-Producing Citrobacter spp. (2015-2017).

The emergence of carbapenem resistance is a significant public health concern. The rate of infections caused by carbapenemase-producing Citrobacter spp., particularly C. freundii, is increasing. Concomitantly, comprehensive global genomic data on carbapenemase-producing Citrobacter spp. are scarce. We used short read whole-genome sequencing to describe the molecular epidemiology and international distribution of eighty-six carbapenemase-producing Citrobacter spp. obtained from two surveillance programs (2015 to 17). The common carbapenemases were KPC-2 (26%), VIM-1 (17%), IMP-4 (14%) and NDM-1 (10%). C. freundii and C. portucalensis were the principal species. C. freundii consisted of multiple clones obtained mainly from Colombia (with KPC-2), the United States (with KPC-2, -3), and Italy (with VIM-1). Two dominant C. freundii clones were identified: ST98 was linked with blaIMP-8 from Taiwan and blaKPC-2 from the United States, and ST22 was linked with blaKPC-2 from Colombia and blaVIM-1 from Italy. C. portucalensis consisted mainly of two clones: ST493 with blaIMP-4 which was limited to Australia, and ST545 with blaVIM-31 which was limited to Turkey. Class I integron (In916) with blaVIM-1 was circulating between multiple sequence types (STs) in Italy, Poland, and Portugal. In73 with blaIMP-8 was circulating between various STs in Taiwan, while In809 with blaIMP-4 was circulating between different STs in Australia. The global carbapenemase-producing Citrobacter spp. population is dominated by diverse STs with different characteristics and varied geographical distribution and thus requires continued monitoring. Ongoing genomic surveillance should use methodologies able to distinguish between C. freundii and C. portucalensis. IMPORTANCE Citrobacter spp. are gaining recognition as important causes of hospital-acquired infections in humans. Among Citrobacter spp., carbapenemase-producing strains are cause of utmost concern to health care services globally due to their ability to resist therapy with virtually any beta-lactam antibiotic. Here, we described the molecular characteristics of a global collection of carbapenemase-producing Citrobacter spp. C. freundii and C. portucalensis were the most common species among Citrobacter spp. with carbapenemases from this survey. Importantly, C. portucalensis was misidentified as C. freundii when using Vitek 2.0/MALDI-TOF MS (matrix-assisted laser desorption/ionization-time of flight mass spectrometry) phenotypic identification, which has important implications for future surveys. Among C. freundii, we identified two dominant clones: ST98 with blaIMP-8 from Taiwan and blaKPC-2 from the United States, and ST22 with blaKPC-2 from Colombia and blaVIM-1 from Italy. As for C. portucalensis, the dominant clones consisted of ST493 with blaIMP-4 from Australia and ST545 with blaVIM-31 from Turkey.

A scoping review of the testing of bulk milk to detect infectious diseases of dairy cattle: Diseases caused by bacteria.

Testing of bulk milk (BM) samples is a convenient, cost-effective strategy that can easily be implemented as part of disease surveillance programs on dairy farms. Here, we performed a scoping review to summarize the literature reporting on the testing of BM samples to detect infectious diseases of dairy cattle caused by bacteria. We also provide a non-exhaustive, albeit significant, list of diagnostic tests that are marketed for BM samples, as well as a list of disease surveillance activities that included testing of BM samples. A literature search was carried out in 5 databases, yielding 8,829 records from which 474 were retained. Overall, 575 eligible bacterial pathogens were screened for using BM samples, ranging from 1 to 6 individual pathogens per study. Staphylococcus aureus, including methicillin-resistant Staph. aureus, were the most studied bacteria (n = 179 studies), followed by Streptococcus agalactiae (86), Mycobacterium avium ssp. paratuberculosis (79), Coxiella burnetii (79), and Mycoplasma spp. (67). Overall, culture-based protocols, ELISA, real-time PCR, and PCR were the most commonly adopted methodologies to screen BM samples. Sensitivity of BM testing for bovine paratuberculosis was generally low and varied greatly according to the ELISA cut-offs adopted and herd-level definition of disease. In general, protocols had low to moderate sensitivities (<50%), which increased for herds with high within-herd seroprevalence. Specificity of BM testing for paratuberculosis was generally high. With respect to mastitis pathogens, BM testing demonstrated high sensitivity and specificity for Strep. agalactiae, in general. However, we observed inconsistency among studies with respect to the sensitivity of BM culture to detect infected herds, which was notably higher if enrolled herds were heavily infected or had history of clinical disease. Among Salmonella spp. pathogens, Salmonella Dublin was the most frequently studied bacterium for which BM testing has been validated. Specificity of BM ELISA was high, ranging from 89.0 to 99.4. In contrast, sensitivity varied greatly among studies, ranging from 50.6% to 100%. Our findings support that one of most important factors affecting sensitivity of BM ELISA for Salmonella Dublin is whether nonlactating cattle are considered in the definition of herd infection status. In general, protocols analyzed in this review suffered from very low sensitivities, which hardly justifies their use as part of disease surveillance as single testing. Nevertheless, test sensitivity can be increased by the adoption of more inclusive definitions of disease-free herds. Further, low-sensitivity and high-specificity methods can be valuable tools for surveillance when used repeatedly over time.

Improved mineralization of dental enamel by electrokinetic delivery of F- and Ca2+ ions.

This in vitro study evaluated the effects of the infiltration of F- and Ca2+ ions into human enamel by electrokinetic flow (EKF) on the enamel microhardness and F- content. Sound human enamel ground sections of unerupted third molars were infiltrated with de-ionized water by EKF and with F- ion by EKF respectively. All samples were submitted to two successive transverse acid-etch biopsies (etching times of 30 s and 20 min) to quantify F- ion infiltrated deep into enamel. Remarkably, sound enamel showed a large increase in microhardness (MH) after infiltration of NaF (p < 0.00001) and CaCl2 (p = 0.013) by EKF. Additionally, NaF-EKF increased the remineralization in the lesion body of artificial enamel caries lesions compared to controls (p < 0.01). With the enamel biopsy technique, at both etching times, more F- ions were found in the EKF-treated group than the control group (p << 0.05), and more fluoride was extracted from deeper biopsies in the NaF-EKF group. In conclusion, our results show that EKF treatment is superior in transporting Ca2+ and F- ions into sound enamel when compared to molecular diffusion, enhancing both the mineralization of sound enamel and the remineralization of artificial enamel caries.

Escherichia coli sequence type 73 bloodstream infections in a centralized Canadian region and their association with companion animals: an ecological study.

PURPOSE: Extraintestinal pathogenic E. coli (ExPEC) are important pathogens causing community-acquired infections in humans, including bloodstream infections (BSIs), and may also colonize and infect animals. Our aim was to investigate associations between incidence rates (IRs) of BSIs caused by ExPEC and number of dogs and cats in communities in Calgary. METHODS: We used a well-characterized collection of blood isolates (n = 685) from Calgary, Alberta, Canada (2016). We used a combination of a seven-single-nucleotide-polymorphism quantitative PCR to type ExPEC into sequence types (STs). Calgary census data were used to estimate IRs per city community, as well as to investigate associations between number of companion animals per community, as obtained from licensing data, and IR of BSIs caused by each dominant ST. RESULTS: From the 685 isolates available, ExPEC ST131 was most prevalent (21.3% of included isolates), followed by ST73 (13.7%), ST69 (8.2%), ST95 (6.7%), and ST1193 (5.3%), respectively. Incidence of BSIs caused by ExPECs among Calgary residents was 48.8 cases per 100,000 resident-years, whereas communities had on average of 1.7 companion animals per 10 residents. No association between the number of dogs and IR of BSIs caused by ExPECs was detected for any ST. Conversely, the incidence rate of BSIs caused by ST73 was 3.6 times higher (95%CI 1.3-9.99) for every increase of 1 cat per 10 habitants in communities. CONCLUSIONS: Number of cats per habitant was positively associated with the incidence of BSIs caused by ExPEC ST73.

Genomic Epidemiology of Global Carbapenemase-Producing Escherichia coli, 2015-2017.

We describe the global molecular epidemiology of 229 carbapenemase-producing Escherichia coli in 36 countries during 2015-2017. Common carbapenemases were oxacillinase (OXA) 181 (23%), New Delhi metallo-ß-lactamase (NDM) 5 (20%), OXA-48 (17%), Klebsiella pneumoniae carbapenemase 2 (15%), and NDM-1 (10%). We identified 5 dominant sequence types (STs); 4 were global (ST410, ST131, ST167, and ST405), and 1 (ST1284) was limited to Turkey. OXA-181 was frequent in Jordan (because of the ST410-B4/H24RxC subclade) and Turkey (because of ST1284). We found nearly identical IncX3-blaOXA-181 plasmids among 11 STs from 12 countries. NDM-5 was frequent in Egypt, Thailand (linked with ST410-B4/H24RxC and ST167-B subclades), and Vietnam (because of ST448). OXA-48 was common in Turkey (linked with ST11260). Global K. pneumoniae carbapenemases were linked with ST131 C1/H30 subclade and NDM-1 with various STs. The global carbapenemase E. coli population is dominated by diverse STs with different characteristics and varied geographic distributions, requiring ongoing genomic surveillance.

Population-based surveillance of Enterobacter cloacae complex causing blood stream infections in a centralized Canadian region.

Active population-based surveillance determined clinical factors, susceptibility patterns, incidence rates (IR), and genomics among Enterobacter cloacae complex (n = 154) causing blood stream infections in a centralized Canadian region (2015-2017). The annual population IR was 1.2/100,000 (95% CI 0.9-16) in 2015, 1.4/100,000 (95% CI 1.1-1.9) in 2016, and 1.5/100,000 (95% CI 1.2-2.0) in 2017, affecting mainly elderly males with underlying comorbid conditions in the hospital setting. E. cloacae complex was dominated by polyclonal subspecies (i.e., E. hormaechei subsp. steigerwaltii, subsp. hoffmanni and subsp. xiangfangesis). Antimicrobial resistant determinants were rare. This study provided novel information about Enterobacter genomics in a well-defined human population.

Knowledge Gaps in the Understanding of Antimicrobial Resistance in Canada.

Current limitations in the understanding and control of antimicrobial resistance (AMR) in Canada are described through a comprehensive review focusing on: (1) treatment optimization; (2) surveillance of antimicrobial use and AMR; and (3) prevention of transmission of AMR. Without addressing gaps in identified areas, sustained progress in AMR mitigation is unlikely. Expert opinions and perspectives contributed to prioritizing identified gaps. Using Canada as an example, this review emphasizes the importance and necessity of a One Health approach for understanding and mitigating AMR. Specifically, antimicrobial use in human, animal, crop, and environmental sectors cannot be regarded as independent; therefore, a One Health approach is needed in AMR research and understanding, current surveillance efforts, and policy. Discussions regarding addressing described knowledge gaps are separated into four categories: (1) further research; (2) increased capacity/resources; (3) increased prescriber/end-user knowledge; and (4) policy development/enforcement. This review highlights the research and increased capacity and resources to generate new knowledge and implement recommendations needed to address all identified gaps, including economic, social, and environmental considerations. More prescriber/end-user knowledge and policy development/enforcement are needed, but must be informed by realistic recommendations, with input from all relevant stakeholders. For most knowledge gaps, important next steps are uncertain. In conclusion, identified knowledge gaps underlined the need for AMR policy decisions to be considered in a One Health framework, while highlighting critical needs to achieve realistic and meaningful progress.

Prevalence, Risk Factors, and Antimicrobial Resistance Profile of Respiratory Pathogens Isolated From Suckling Beef Calves to Reprocessing at the Feedlot: A Longitudinal Study.

Here, we investigated the prevalence and risk factors for the presence of Histophilus somni, Mannheimia haemolytica, Mycoplasma bovis, and Pasteurella multocida in the respiratory tract of calves from the spring processing to the reprocessing at feedlots. Additionally, we characterized, phenotypically and genotypically, the antimicrobial resistance (AMR) profile of the four species. Calves from 22 cow-calf operations were enrolled in the study (n = 30 calves per operation) and sampled by deep nasopharyngeal swabs at three time points: spring processing, weaning, or induction into feedlots, and at reprocessing at the feedlot. Isolates were tested for susceptibility using the minimum inhibitory concentration (MIC) test against commonly administered antimicrobials. Additionally, a subset of isolates underwent whole-genome sequencing to infer presence of AMR genes and resistance determinants. Among studied pathogens, P. multocida was the most prevalent species, regardless of time point, followed by M. haemolytica, M. bovis, and H. somni. For M. bovis, a sharp increase in prevalence was detected at the reprocessing sampling, whereas for P. multocida, an increase in prevalence was observed at the weaning/induction sampling. Comingling and co-location of feedlots were not associated with prevalence of any respiratory pathogen. In terms of AMR, resistance against macrolides was prevalent in M. bovis, with most isolates resistant against tildipirosin, tilmicosin, and tylosin. In general, there was limited evidence to support an increase in resistance rates of respiratory bacteria from the spring processing to reprocessing at feedlots, with the exception of florfenicol resistance in M. bovis, which increased at reprocessing. Metaphylactic administration of tetracyclines at feedlot induction was not associated with the MIC of tetracyclines in any respiratory bacteria. Conversely, there were clear associations between the parenteral use of macrolides as metaphylaxis at the feedlot induction, and increased MIC against macrolides in P. multocida, M. haemolytica, and H. somni. Overall, the AMR phenotypes were corroborated by presence of AMR genes. We hypothesize that the administration of macrolides such as tulathromycin at feedlot induction contributes to historical changes in macrolides MIC data of respiratory bacteria of beef cattle.