Multidrug-Resistant Klebsiella variicola Isolated in the Urine of Healthy Bovine Heifers, a Potential Risk as an Emerging Human Pathogen.

Klebsiella variicola, a member of Klebsiella pneumoniae complex, is found to infect plants, insects, and animals and is considered an emerging pathogen in humans. While antibiotic resistance is often prevalent among K. variicola isolates from humans, this has not been thoroughly investigated in isolates from nonhuman sources. Prior evidence suggests that K. variicola can be transmitted between agricultural products as well as between animals, and the use of antibiotics in agriculture has increased antibiotic resistance in other emerging pathogens. Furthermore, in animals that contain K. variicola as a normal member of the rumen microbiota, the same bacteria can also cause infections, such as clinical mastitis in dairy cows. Here, we describe K. variicola UFMG-H9 and UFMG-H10, both isolated from the urine of healthy Gyr heifers. These two genomes represent the first isolates from the urine of cattle and exhibit greater similarity with strains from the human urinary tract than isolates from bovine fecal or milk samples. Unique to the UFMG-H9 genome is the presence of flagellar genes, the first such observation for K. variicola. Neither of the sampled animals had symptoms associated with K. variicola infection, even though genes associated with virulence and antibiotic resistance were identified in both strains. Both strains were resistant to amoxicillin, erythromycin, and vancomycin, and UFMG-H10 is resistant to fosfomycin. The observed resistances emphasize the concern regarding the emergence of this species as a human pathogen given its circulation in healthy livestock animals. IMPORTANCE Klebsiella variicola is an opportunistic pathogen in humans. It also has been associated with bovine mastitis, which can have significant economic effects. While numerous isolates have been sequenced from human infections, only 12 have been sequenced from cattle (fecal and milk samples) to date. Recently, we discovered the presence of K. variicola in the urine of two healthy heifers, the first identification of K. variicola in the bovine urinary tract and the first confirmed K. variicola isolate encoding for flagella-mediated motility. Here, we present the genome sequences and analysis of these isolates. The bovine urinary genomes are more similar to isolates from the human urinary tract than they are to other isolates from cattle, suggesting niche specialization. The presence of antibiotic resistance genes is concerning, as prior studies have found transmission between animals. These findings are important to understand the circulation of K. variicola in healthy livestock animals.

Escherichia coli and Pseudomonas aeruginosa Isolated From Urine of Healthy Bovine Have Potential as Emerging Human and Bovine Pathogens.

The study of livestock microbiota has immediate benefits for animal health as well as mitigating food contamination and emerging pathogens. While prior research has indicated the gastrointestinal tract of cattle as the source for many zoonoses, including Shiga-toxin producing Escherichia coli and antibiotic resistant bacteria, the bovine urinary tract microbiota has yet to be thoroughly investigated. Here, we describe 5 E. coli and 4 Pseudomonas aeruginosa strains isolated from urine of dairy Gyr cattle. While both species are typically associated with urinary tract infections and mastitis, all of the animals sampled were healthy. The bovine urinary strains were compared to E. coli and P. aeruginosa isolates from other bovine samples as well as human urinary samples. While the bovine urinary E. coli isolates had genomic similarity to isolates from the gastrointestinal tract of cattle and other agricultural animals, the bovine urinary P. aeruginosa strains were most similar to human isolates suggesting niche adaptation rather than host adaptation. Examination of prophages harbored by these bovine isolates revealed similarity with prophages within distantly related E. coli and P. aeruginosa isolates from the human urinary tract. This suggests that related urinary phages may persist and/or be shared between mammals. Future studies of the bovine urinary microbiota are needed to ascertain if E. coli and P. aeruginosa are resident members of this niche and/or possible sources for emerging pathogens in humans.

Corynebacterium phoceense, resident member of the urogenital microbiota?

Corynebacterium phoceense is a Gram-positive species previously isolated from human urine. Although other species from the same genus have been associated with urinary tract infections, C. phoceense is currently believed to be a non-pathogenic member of the urogenital microbiota. Prior to our study, only two isolates were described in the literature, and very little is known about the species. Here, we describe C. phoceense UFMG-H7, the first strain of this species isolated from the urine of healthy cattle. The genome for this isolate was produced and compared to the two other publicly available C. phoceense as well as other Corynebacterium genome assemblies. Our in-depth genomic analysis identified four additional publicly available genome assemblies that are representatives of the species, also isolated from the human urogenital tract. Although none of the strains have been associated with symptoms or disease, numerous genes associated with virulence factors are encoded. In contrast to related Corynebacterium species and Corynebacterium species from the bovine vaginal tract, all C. phoceense strains examined code for the SpaD-type pili suggesting adherence is essential for its persistence within the urinary tract. As the other C. phoceense strains analysed were isolated from the human urogenital tract, our results suggest that this species may be specific to this niche.

Examination of Staphylococcus aureus Prophages Circulating in Egypt.

Staphylococcus aureus infections are of growing concern given the increased incidence of antibiotic resistant strains. Egypt, like several other countries, has seen alarming increases in methicillin-resistant S. aureus (MRSA) infections. This species can rapidly acquire genes associated with resistance, as well as virulence factors, through mobile genetic elements, including phages. Recently, we sequenced 56 S. aureus genomes from Alexandria Main University Hospital in Alexandria, Egypt, complementing 17 S. aureus genomes publicly available from other sites in Egypt. In the current study, we found that the majority (73.6%) of these strains contain intact prophages, including Biseptimaviruses, Phietaviruses, and Triaviruses. Further investigation of these prophages revealed evidence of horizontal exchange of the integrase for two of the prophages. These Egyptian S. aureus prophages are predicted to encode numerous virulence factors, including genes associated with immune evasion and toxins, including the Panton-Valentine leukocidin (PVL)-associated genes lukF-PV/lukS-PV. Thus, prophages are likely to be a major contributor to the virulence of S. aureus strains in circulation in Egypt.

Vagococcus fluvialis isolation and sequencing from urine of healthy cattle.

While the gram-positive bacterium Vagococcus fluvialis has been isolated from the environment as well as fish, birds, and mammals, very little is known about the species. V. fluvialis is believed to be a probiotic in fishes. However, within mammals, it is more frequently isolated from infectious tissue, including on rare occasions human and livestock lesions. Prior to the study described here, V. fluvialis had never been found in healthy bovine animals. Here, we present the complete genomes of V. fluvialis UFMG-H6, UFMG-H6B, and UFMG-H7, novel strains isolated from urine samples from healthy bovine females. These are the first genomes of mammalian isolates and the first description of V. fluvialis from urine. The genomes did not encode for any known virulence genes, suggesting that they may be commensal members of the urine microbiota.

Draft Genome Sequence of Citrobacter freundii UFMG-H9, Isolated from Urine from a Healthy Bovine Heifer (Gyr Breed).

Citrobacter freundii is a pathogen associated with antibiotic resistance and severe infections in humans. Here, we report the draft genome sequence of C. freundii strain UFMG-H9, an isolate from urine from a healthy Gyr heifer.

Draft Genome Sequence of Aeromonas caviae UFMG-H8, Isolated from Urine from a Healthy Bovine Heifer (Gyr Breed).

Aeromonas caviae is an emerging pathogen in humans, causing intestinal infections. Here, we report Aeromonas caviae strain UFMG-H8, isolated from the urine of a healthy heifer (Gyr breed).

Draft Genome Sequence of Enterobacter asburiae UFMG-H9, Isolated from Urine from a Healthy Bovine Heifer (Gyr Breed).

Enterobacter asburiae is part of the Enterobacter cloacae complex, related to nosocomial opportunistic infections in humans. Here, we report the draft genome of E. asburiae strain UFMG-H9, an isolate from urine from a healthy Gyr heifer.

Draft Genome Sequences of Three Enterococcus casseliflavus Strains Isolated from the Urine of Healthy Bovine Heifers (Gyr Breed).

Enterococcus casseliflavus is a commensal bacterium present in the intestinal microbiota of different animals. Previous studies have found that strains isolated from livestock are often resistant to many different antibiotics. Here, we present three E. casseliflavus strains, UFMG-H7, UFMG-H8, and UFMG-H9, isolated from urine collected from healthy dairy heifers in Brazil.

Three Draft Genome Sequences of Staphylococcus Species Isolated from the Urine of Healthy Bovine Heifers (Gyr Breed).

Members of the Staphylococcus genus are known pathogens causing mastitis in dairy cows, which results in major economic losses. Here, we present Staphylococcus epidermidis UFMG-H7, Staphylococcus hominis UFMG-H7B, and Staphylococcus sciuri UFMG-H6, isolated from the urine of healthy purebred Gyr heifers.

A common vaginal microbiota composition among breeds of Bos taurus indicus (Gyr and Nellore).

Describing the bovine vaginal microbiota is essential to better understand its physiology and its impact on health maintenance. Despite the economic importance of reproduction of these animals, bovine vaginal microbial community is still poorly described in comparison with rumen microbiome. Previous studies of our group described the vaginal microbiota of Nellore, an important Bos taurus indicus breed, using metagenomics. In order to better understand this microbiota, the present work aims to investigate another important breed, Gyr. Results have shown bacterial dominance over Archaea and Fungi was observed, with the most abundant bacterial phylum (Firmicutes) representing 40-50% of bacterial population, followed by Bacteroidetes, Proteobacteria, and Actinobacteria. The Fungi kingdom had the Mycosphaerella genus as its main representative, followed by Cladosporium. Archaea were observed at a very low abundance in all animals, with a high relative abundance of Methanobrevibacter genus. These results demonstrate a high microbial diversity on vaginal tract of Gyr, as demonstrated for Nellore and different from the previously described for other species. Our results indicate a great similarity between vaginal microbiota of Nellore and Gyr despite the differences in animal handling and genetic improvement. As observed for both breeds, individual variation is the largest source of microbial diversity between animals.

Prospective comparative study of endoscopic submucosal dissection and gastrectomy for early neoplastic lesions including patients' perspectives.

BACKGROUND: There are no prospective studies comparing endoscopic submucosal dissection (ESD) and gastrectomy, especially evaluating patient-reported outcomes. Our aim was to compare the safety and impact on quality of life (QoL) of ESD and gastrectomy in patients with early gastric neoplasia. METHODS: This prospective study included consecutive patients presenting with early gastric neoplasia in a tertiary center from January 2015 to August 2016. Data collection included curative resection, adverse events (AEs), and patient-reported outcomes (questionnaires: EORTC QLQ-C30, EORTC STO-22, EQ-5D-5 L, and Assessment of Survivor Concerns) before and after interventions (after 1 month, 3 - 6 months, and 1 year). RESULTS: 254 patients with early lesions were included: 153 managed by ESD and 101 by gastrectomy, the former being significantly older and with less advanced lesions. Mean procedural time and length of stay were significantly higher in the surgery group (164 vs. 72 minutes and 16.3 vs. 3.5 days; P < 0.001). Complete resection was higher in the surgical group (99 % vs. 90 %; P = 0.02); ESD was curative in 79 % of patients. Severe AEs and surgical re-intervention were significantly more frequent in the gastrectomy group (21.8 % vs. 7.8 % and 11 % vs. 1 %, respectively). Endoscopic treatment was associated with a positive impact on global health-related QoL at 1 year (net difference + 9.9; P = 0.006), role function and symptom scales (fatigue, pain, appetite, eating restrictions, dysphagia, and body image). Concerns about recurrence did not differ between the groups. CONCLUSIONS: In patients with early gastric neoplasia, ESD is safer and is associated with a positive impact on health-related QoL when compared with gastrectomy, without increasing fear of recurrence and new lesions.

Vaginal Microbiome Characterization of Nellore Cattle Using Metagenomic Analysis.

Understanding of microbial communities inhabiting cattle vaginal tract may lead to a better comprehension of bovine physiology and reproductive health being of great economic interest. Up to date, studies involving cattle microbiota are focused on the gastrointestinal tract, and little is known about the vaginal microbiota. This study aimed to investigate the vaginal microbiome in Nellore cattle, heifers and cows, pregnant and non-pregnant, using a culture independent approach. The main bacterial phyla found were Firmicutes (~40-50%), Bacteroidetes (~15-25%) and Proteobacteria (~5-25%), in addition to ~10-20% of non-classified bacteria. 45-55% of the samples were represented by only ten OTUs: Aeribacillus, Bacteroides, Clostridium, Ruminococcus, Rikenella, Alistipes, Bacillus, Eubacterium, Prevotella and non-classified bacteria. Interestingly, microbiota from all 20 animals could be grouped according to the respiratory metabolism of the main OTUs found, creating three groups of vaginal microbiota in cattle. Archaeal samples were dominated by the Methanobrevibacter genus (Euryarchaeota, ~55-70%). Ascomycota was the main fungal phylum (~80-95%) and Mycosphaerella the most abundant genus (~70-85%). Hormonal influence was not clear, but a tendency for the reduction of bacterial and increase of archaeal populations in pregnant animals was observed. Eukaryotes did not vary significantly between pregnant and non-pregnant animals, but tended to be more abundant on cows than on heifers. The present work describes a great microbial variability in the vaginal community among the evaluated animals and groups (heifers and cows, pregnant and non-pregnant), which is significantly different from the findings previously reported using culture dependent methods, pointing out the need for further studies on this issue. The microbiome found also indicates that the vaginal colonization appears to be influenced by the gastrointestinal community.

Epidemiology of total hip and knee replacement: a cross-sectional study.

OBJECTIVE: To describe the epidemiologic characteristics and adverse events of patients submitted to total hip and total knee replacement. METHODS: A cross-sectional study retrospectively assessing medical chart data of all total hip and total knee replacements performed at a private hospital, between January 2007 and December 2010 Patients submitted to total hip and total knee replacement, with consent of surgeons were included. Incomplete records and/or missing data of the hospital database were excluded. The categorical variables analyzed were age, gender, type of arthroplasty (primary or secondary), type of procedure, duration of surgery, use of drains, risk of infection, compliance to protocol for prevention of deep venous thrombosis and embolism pulmonary, and compliance to the protocol for prevention of infection. The outcomes assessed were adverse events after surgery. RESULTS: A total of 510 patients were included; in that, 166 admissions for knee replacements (92 male) and 344 admissions for hip replacements (176 female). The mean age of patients was 71 years (range 31-99 years). Adverse events were reported in 76 patients (14.9%); there was no correlation between assessed variables and number of complications. CONCLUSION: The results showed no individual factors favoring complications in patients submitted to total hip and total knee replacement; hence, surgeons should consider prophylaxis to avoid complications.

Epidemiology of total hip and knee replacement: a cross-sectional study / Epidemiologia da artroplastia total de quadril e de joelho: estudo transversal

OBJECTIVE: To describe the epidemiologic characteristics and adverse events of patients submitted to total hip and total knee replacement. METHODS: A cross-sectional study retrospectively assessing medical chart data of all total hip and total knee replacements performed at a private hospital, between January 2007 and December 2010 Patients submitted to total hip and total knee replacement, with consent of surgeons were included. Incomplete records and/or missing data of the hospital database were excluded. The categorical variables analyzed were age, gender, type of arthroplasty (primary or secondary), type of procedure, duration of surgery, use of drains, risk of infection, compliance to protocol for prevention of deep venous thrombosis and embolism pulmonary, and compliance to the protocol for prevention of infection. The outcomes assessed were adverse events after surgery. RESULTS: A total of 510 patients were included; in that, 166 admissions for knee replacements (92 male) and 344 admissions for hip replacements (176 female). The mean age of patients was 71 years (range 31-99 years). Adverse events were reported in 76 patients (14.9%); there was no correlation between assessed variables and number of complications. CONCLUSION: The results showed no individual factors favoring complications in patients submitted to total hip and total knee replacement; hence, surgeons should consider prophylaxis to avoid complications.

OBJETIVO: Descrever as características epidemiológicas e os eventos adversos dos pacientes submetidos aos procedimentos de artroplastia de quadril e joelho. MÉTODOS: Estudo transversal, com avaliação retrospectiva de prontuários médicos de todas artroplastias de quadril e de joelho realizadas entre janeiro de 2007 e dezembro de 2010, em hospital privado. Foram incluídos pacientes adultos submetidos à artroplastia total do quadril ou do joelho, com anuência dos cirurgiões. Foram excluídos os pacientes com prontuários incompletos ou perdidos na base de dados do hospital. As variáveis categóricas analisadas foram: idade, gênero, tipo de artroplastia (primária ou secundária), tipo de procedimento, tempo de cirurgia, uso de drenos, risco de infecção, adesão ao protocolo para prevenção de trombose venosa profunda e embolia pulmonar, e adesão ao protocolo para prevenção de infecção. Os desfechos avaliados dos pacientes foram eventos adversos ocorridos após a cirurgia. RESULTADOS: Foram incluídos 510 pacientes, sendo 166 hospitalizações para artroplastia do joelho (92 do gênero masculino) e 344 hospitalizações para artroplastia do quadril (176 do gênero feminino). A média da idade dos pacientes foi 71 anos (variação 31 a 99 anos). Os eventos adversos ocorreram em 76 pacientes (14,9%); não foi encontrada correlação entre as variáveis avaliadas e o número de complicações. CONCLUSÃO: Os dados não demonstraram fatores individuais que favorecessem complicações de pacientes submetidos à artroplastia do quadril e joelho; portanto, os cirurgiões têm que considerar administrações profiláticas para evitar complicações.

Platelet-rich plasma for long bone healing.

OBJECTIVE: To evaluate effectiveness of the use of platelet-rich plasma as coadjuvant for union of long bones. METHODS: The search strategy included the Cochrane Library (via Central) and MEDLINE (via PubMed). There were no limits as to language or publication media. The latest search strategy was conducted in December 2011. It included randomized clinical trials that evaluated the use of platelet-rich plasma as coadjuvant medication to accelerate union of long bones (acute fractures, pseudoarthrosis and bone defects). The outcomes of interest for this review include bone regeneration, adverse events, costs, pain, and quality of life. The authors selected eligible studies, evaluated the methodological quality, and extracted the data. It was not possible to perform quantitative analysis of the grouped studies (meta-analyses). RESULTS: Two randomized prospective clinical trials were included, with a total of 148 participants. One of them compared recombinant human morphogenic bone protein-7 versus platelet-rich plasma for the treatment of pseudoarthrosis; the other evaluated the effects of three coadjuvant treatments for union of valgising tibial osteotomies (platelet-rich plasma, platelet-rich plasma plus bone marrow stromal cells, and no coadjuvant treatment). Both had low statistical power and moderate to high risk of bias. CONCLUSION: There was no conclusive evidence that sustained the use of platelet-rich plasma as a coadjuvant to aid bone regeneration of fractures, pseudoarthrosis, or bone defects.

Plasma rico em plaquetas para consolidação de ossos longos / Platelet-rich plasma for long bone healing

OBJETIVO: Avaliar a efetividade do uso de plasma rico em plaquetas como coadjuvante para a consolidação óssea de ossos longos. MÉTODOS: A estratégia de busca abrangeu a Cochrane Library (via Central) e o MEDLINE (via PubMed). Não houve restrições de idioma ou meios de publicações. A última estratégia de busca foi realizada em dezembro de 2011. Foram incluídos ensaios clínicos randomizados que avaliaram o uso do plasma rico em plaquetas como medicação coadjuvante para acelerar a consolidação dos ossos longos (fraturas agudas, pseudoartroses e defeitos ósseos). Os desfechos de interesse para esta revisão compreenderam: consolidação óssea, eventos adversos, custos, dor e qualidade de vida. Os autores selecionaram os estudos elegíveis, avaliaram a qualidade metodológica e extraíram os dados. Não foi possível realizar análise quantitativa dos estudos agrupados (meta-análises). RESULTADOS: Foram incluídos dois ensaios clínicos prospectivos randomizados, envolvendo um total de 148 participantes. Um deles comparou proteína morfogenética óssea recombinante humana 7 versus PRP para o tratamento de pseudoartroses; o outro avaliou os efeitos de três tratamentos coadjuvantes para a consolidação de osteotomias valgizantes da tíbia (plasma rico em plaquetas, plasma rico em plaquetas mais células estromais da medula óssea e sem tratamento coadjuvante). Ambos possuíam baixo poder estatístico e moderado a alto risco de viés. CONCLUSÃO: Não houve evidências conclusivas que sustentassem o uso de plasma rico em plaquetas como coadjuvante para auxiliar a consolidação óssea de fraturas, pseudoartrose ou defeitos ósseos.

OBJECTIVE: To evaluate effectiveness of the use of platelet-rich plasma as coadjuvant for union of long bones. METHODS: The search strategy included the Cochrane Library (via Central) and MEDLINE (via PubMed). There were no limits as to language or publication media. The latest search strategy was conducted in December 2011. It included randomized clinical trials that evaluated the use of platelet-rich plasma as coadjuvant medication to accelerate union of long bones (acute fractures, pseudoarthrosis and bone defects). The outcomes of interest for this review include bone regeneration, adverse events, costs, pain, and quality of life. The authors selected eligible studies, evaluated the methodological quality, and extracted the data. It was not possible to perform quantitative analysis of the grouped studies (meta-analyses). RESULTS: Two randomized prospective clinical trials were included, with a total of 148 participants. One of them compared recombinant human morphogenic bone protein-7 versus platelet-rich plasma for the treatment of pseudoarthrosis; the other evaluated the effects of three coadjuvant treatments for union of valgising tibial osteotomies (platelet-rich plasma, platelet-rich plasma plus bone marrow stromal cells, and no coadjuvant treatment). Both had low statistical power and moderate to high risk of bias. CONCLUSION: There was no conclusive evidence that sustained the use of platelet-rich plasma as a coadjuvant to aid bone regeneration of fractures, pseudoarthrosis, or bone defects.