Proximal Femur Endoprosthesis Augmentation with Polymethylmethacrylate and Condylar Plate: A Case Series.

Introduction: Aseptic and septic loosening by mechanical failure is one of the main causes of proximal femur endoprosthesis failure with different clinical consequences such as pain, inflammation, progressive loosening of muscular volume, and strength with functional limitation. We present a case series of four patients with aseptic and septic loosening of femur endoprosthesis that was treated with a novel technique that combines different methods to achieve primary stability. Case Report: Four patients with an average age of 49.5 years (35-70 years), two women and two men, were referred to the Orthopedic Surgery Department of the San Ignacio University Hospital of Bogota. Two of them with a history of oncological disease, ruling out a tumor relapse through local images and extension examinations; another patient with a history of firearm injury to the left thigh with multiple secondary complications, which required various reconstructive procedures; and the oldest patient with a history of total hip replacement. The main complaint of the four patients was thigh pain not caused by trauma and initial assessment radiographs of all of them showed signs of loosening of the prosthetic material, with septic etiology in two of the patients and aseptic etiology in the other two. Therefore, they were scheduled for revision surgery of prosthetic components. These patients recover functional status and resolved pain in the early post-operative period and during long-term follow-up. Only one of the patients, time after the surgical procedure, required removal of the osteosynthesis material due to persistent bone infectious process. Conclusion: Septic and aseptic loosening is one of the most common complications of proximal femur endoprosthesis, resulting in significant pain and functional decline in patients. We present a novel surgical technique that allows primary stabilization of the construct that allows early rehabilitation, improvement of functionality, and no signs of new loosening.

Evaluation of immunogenicity and efficacy of the enterobactin conjugate vaccine in protecting chickens from colibacillosis.

Colibacillosis is one of the most common and economically devastating infectious diseases in poultry production worldwide. Innovative universal vaccines are urgently needed to protect chickens from the infections caused by genetically diverse avian pathogenic Escherichia coli (APEC). Enterobactin (Ent) is a highly conserved siderophore required for E. coli iron acquisition and pathogenesis. The Ent-specific antibodies induced by a novel Ent conjugate vaccine significantly inhibited the in vitro growth of diverse APEC strains. In this study, White Leghorn chickens were immunized with the Ent conjugate vaccine using a crossed design with two variables, vaccination (with or without) and APEC challenge (O1, O78, or PBS control), resulting in six study groups (9 to 10 birds/group). The chickens were subcutaneously injected with the vaccine (100 µg per bird) at 7 days of age, followed by booster immunization at 21 days of age. The chickens were intratracheally challenged with an APEC strain (108 CFU/bird) or PBS at 28 days of age. At 5 days post infection, all chickens were euthanized to examine lesions and APEC colonization of the major organs. Immunization of chickens with the Ent vaccine elicited a strong immune response with a 64-fold increase in the level of Ent-specific IgY in serum. The hypervirulent strain O78 caused extensive lesions in lung, air sac, heart, liver, and spleen with significantly reduced lesion scores observed in the vaccinated chickens. Interestingly, the vaccination did not significantly reduce APEC levels in the examined organs. The APEC O1 with low virulence only caused sporadic lesions in the organs in both vaccination and control groups. The Ent conjugate vaccine altered the bacterial community of the ileum and cecum. Taken together, the findings from this study showed the Ent conjugate vaccine could trigger a strong specific immune response and was promising to confer protection against APEC infection.

Identification of novel genes involved in the biofilm formation process of Avian Pathogenic Escherichia coli (APEC).

Avian pathogenic Escherichia coli (APEC) is the etiological agent of avian colibacillosis, a leading cause of economic loss to the poultry industry worldwide. APEC causes disease using a diverse repertoire of virulence factors and has the ability to form biofilms, which contributes to the survival and persistence of APEC in harsh environments. The objective of this study was to identify genes most widespread and important in APEC that contribute to APEC biofilm formation. Using the characterized APEC O18 as the template strain, a total of 15,660 mutants were randomly generated using signature tagged mutagenesis and evaluated for decreased biofilm formation ability using the crystal violet assay. Biofilm deficient mutants were sequenced, and a total of 547 putative biofilm formation genes were identified. Thirty of these genes were analyzed by PCR for prevalence among 109 APEC isolates and 104 avian fecal E. coli (AFEC) isolates, resulting in nine genes with significantly greater prevalence in APEC than AFEC. The expression of these genes was evaluated in the wild-type APEC O18 strain using quantitative real-time PCR (qPCR) in both the exponential growth phase and the mature biofilm phase. To investigate the role of these genes in biofilm formation, isogenic mutants were constructed and evaluated for their biofilm production and planktonic growth abilities. Four of the mutants (rfaY, rfaI, and two uncharacterized genes) displayed significantly decreased biofilm formation, and of those four, one (rfaI) displayed significantly decreased growth compared to the wild type. Overall, this study identified novel genes that may be important in APEC and its biofilm formation. The data generated from this study will benefit further investigation into the mechanisms of APEC biofilm formation.

Necrotizing Hepatitis Associated with Clostridium perfringens in Broiler Chicks.

In this retrospective study we describe unusual cases of clostridial hepatitis associated with high mortality in young broiler chicks. Eleven cases of necrotizing hepatitis in broiler chicks from four companies were submitted to the Poultry Diagnostic and Research Center or the Georgia Poultry Laboratory Network between 2017 and 2020. In most flocks, increased 3-day mortality was followed by an elevated 7-day mortality. Gross lesions included green to dark brown discoloration of the liver, congested lungs, serosanguineous fluid in the caudoventral aspect of the abdomen, and emphysema in the yolk sacs. In birds older than a week of age, disease with neurologic signs became evident and consisted of tremors, stargazing, and incoordination. Histopathologic evaluation revealed multifocal to coalescing fibrinoheterophilic and necrotizing hepatitis associated with gram-positive, long, rod-shaped bacteria. Formalin-fixed liver samples from six cases out of eight cases tested were positive for Clostridium perfringens by immunohistochemistry. Liver samples from two cases were culture positive for Clostridium spp., and C. perfringens was isolated from one sample. Toxinotyping by PCR performed in seven samples revealed the presence of the genes that code for alpha toxin phospholipase C (cpa or plc) and necrotic enteritis toxin B-like (netB) in six samples and as well as C. perfringens large cytotoxin (tpeL) in one sample. Broiler breeders are the suspected source of the infection, and testing revealed C. perfringens in hatchery samples and among broiler breeder flocks. Antimicrobial therapy was coupled with enhanced sanitation at the farm and hatchery in that company, markedly decreasing the mortality and clinical signs. This is the first comprehensive evaluation of clostridial necrotizing hepatitis in newly hatched chicks, and the second ever reported in the literature.

Hepatitis necrotizante asociada con Clostridium perfringens en pollos de engorde En este estudio retrospectivo se describen casos inusuales de hepatitis clostridial asociados con una alta mortalidad en pollos de engorde jóvenes. Once casos de hepatitis necrotizante en pollos de engorde de cuatro empresas se enviaron al Centro de Investigación y Diagnóstico Avícola o a la Red de Laboratorios Avícolas del Estado Georgia entre los años 2017 y 2020. En la mayoría de las parvadas, el aumento de la mortalidad a los tres días fue seguido por una mortalidad elevada a los siete días. Las lesiones macroscópicas incluyeron coloración del hígado de verde a marrón oscuro, pulmones congestionados, líquido serosanguinolento en la cara caudoventral del abdomen y enfisema en los sacos vitelinos. En aves mayores de una semana de edad, la enfermedad con signos neurológicos se hizo evidente y consistía en temblores, torticolis (aves como observando a las estrellas) y falta de coordinación. La evaluación histopatológica reveló hepatitis multifocal a fibrinoheterófila coalescente y necrotizante asociada con bacterias grampositivas largas en forma de bastón. Las muestras de hígado fijadas en formalina de seis casos de los ocho casos analizados dieron positivo para Clostridium perfringens por inmunohistoquímica. Las muestras de hígado de dos casos dieron positivo en cultivo para Clostridium spp., y se aisló C. perfringens de una muestra. La tipificación por el tipo de toxina mediante PCR realizado en siete muestras reveló la presencia de los genes que codifican la toxina alfa fosfolipasa C (cpa, plc) y la toxina de enteritis necrótica similar a la toxina B (netB) en seis muestras, así como la citotoxina grande de C. perfringens (tpeL) en una muestra. Se sospecha que las reproductoras de pollos de engorde son la fuente de la infección, y las pruebas revelaron C. perfringens en las muestras de las incubadoras y entre las parvadas de reproductoras de pollos de engorde. La terapia antimicrobiana se combinó con un saneamiento mejorado en la granja y en la incubadora de esa empresa, lo que redujo notablemente la mortalidad y los signos clínicos. Esta es la primera evaluación exhaustiva de la hepatitis necrosante por clostridios en pollitos recién nacidos y la segunda que se ha informado en la literatura.

Efficacy and utilization of an acceptance and commitment therapy-based smartphone application for smoking cessation among Black adults: secondary analysis of the iCanQuit randomized trial.

BACKGROUND AND AIMS: Black adults who smoke are less likely to seek treatment and to succeed in quitting compared with other racial groups. The lack of efficacious and engaging trials for smoking cessation further contributes to this disparity. This study explored whether an acceptance and commitment therapy (ACT)-based smartphone application (iCanQuit) was more efficacious for smoking cessation than a United States Clinical Practice Guidelines (USCPG)-based smartphone application (QuitGuide) among Black adults. DESIGN: Secondary analysis of a two-arm randomized trial with 12-month follow-up. SETTING: United States (US). PARTICIPANTS: A total of 554 Black adults who smoke daily were recruited from 34 US states and enrolled between May 2017 and September 2018. INTERVENTIONS: Participants were randomized to receive iCanQuit (n = 274) or QuitGuide (n = 280) for 12 months. MEASUREMENTS: Smoking cessation outcomes were measured at 3, 6, and 12 months. The primary outcome was self-reported complete-case 30-day point prevalence abstinence (PPA) at 12 months. Secondary outcomes were 7-day PPA, missing-as-smoking imputation, multiple imputation, prolonged abstinence, and cessation of all tobacco products at 12 months. Study retention, treatment engagement, and change in ACT-based processes were also compared between arms. FINDINGS: Study retention was 89% at 12 months and did not differ by arm (P > 0.05). The complete-case 30-day PPA was 28% for iCanQuit versus 20% for QuitGuide at 12 months (odds ratio [OR] = 1.60; 95% confidence interval [CI] = 1.03, 2.46). Similar associations were observed for the missing-as-smoking imputation, although non-significant (25% iCanQuit vs 18% QuitGuide; OR = 1.50; 95% CI = 0.98, 2.30). iCanQuit vs QuitGuide participants were significantly more engaged with iCanQuit application as measured by the number of logins from baseline to 6 months (incidence rate ratio = 3.26; 95% CI = 2.58, 4.13). Increased acceptance of cues to smoke mediated the effect of treatment on cessation (indirect effect: OR = 0.20; 95% CI = 0.05, 0.29). CONCLUSIONS: Among Black adults, an acceptance and commitment therapy-based smartphone application appeared to be more efficacious and engaging for smoking cessation than the United States Clinical Practice Guidelines-based QuitGuide application.

Severe Acute Respiratory Syndrome Coronavirus 2: Vaccine Hesitancy Among Underrepresented Racial and Ethnic Groups With HIV in Miami, Florida.

BACKGROUND: Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and human immunodeficiency virus (HIV) disproportionally affect underrepresented ethnoracial groups in the United States. Medical mistrust and vaccine hesitancy will likely impact acceptability of SARS-CoV-2 vaccines. This study examined SARS-CoV-2 vaccine hesitancy among underrepresented ethnoracial groups with HIV and identified factors that may reduce vaccine uptake. METHODS: We conducted a cross-sectional study of adults ≥18 years of age with HIV residing in Miami, Florida. Participants were invited to participate in the ACTION (A Comprehensive Translational Initiative on Novel Coronavirus) cohort study. A baseline survey was administered from April to August 2020 and followed by a coronavirus disease 2019 (COVID-19) vaccine hesitancy survey from August to November 2020. The COVID-19 vaccine hesitancy survey was adapted from the Strategic Advisory Group of Experts survey. Comparisons by race and ethnicity were performed using the Freedman-Haltmann extension of the Fisher exact test. RESULTS: A total of 94 participants were enrolled; mean age was 54.4 years, 52% were female, 60% were Black non-Latinx, and 40% were non-Black Latinx. Black non-Latinx participants were less likely to agree that vaccinations are important for health when compared to non-Black Latinx (67.8% vs 92.1%, P = .009), less likely to agree that vaccines are effective in preventing disease (67.8% vs 84.2%, P = .029), less likely to believe that vaccine information is reliable and trustworthy (35.7% vs 71.1%, P = .002), and less likely to believe vaccines were unnecessary because COVID-19 would disappear soon (11% vs 21%, P = .049). CONCLUSIONS: Medical mistrust, vaccine hesitancy, and negative sentiments about SARS-CoV-2 vaccines are prevalent among underrepresented ethnoracial groups with HIV, particularly Black non-Latinx. Targeted strategies to increase vaccine uptake in this population are warranted.

Complete Genome Sequence of the Neonatal Meningitis Escherichia coli Serotype O18:K1 Strain NMEC15.

Neonatal meningitis Escherichia coli (NMEC) is the second leading cause of sepsis and meningitis in neonates worldwide. Here, we report the genome sequence of NMEC15, belonging to serotype O18:K1, isolated from the cerebrospinal fluid (CSF) of an infant with neonatal bacterial meningitis (NBM) in the Netherlands.

Evaluation of the Immunogenic Response of a Novel Enterobactin Conjugate Vaccine in Chickens for the Production of Enterobactin-Specific Egg Yolk Antibodies.

Passive immunization with specific egg yolk antibodies (immunoglobulin Y, IgY) is emerging as a promising alternative to antibiotics to control bacterial infections. Recently, we developed a novel conjugate vaccine that could trigger a strong immune response in rabbits directed against enterobactin (Ent), a highly conserved siderophore molecule utilized by different Gram-negative pathogens. However, induction of Ent-specific antibodies appeared to be affected by the choice of animal host and vaccination regimen. It is still unknown if the Ent conjugate vaccine can trigger a specific immune response in layers for the purpose of production of anti-Ent egg yolk IgY. In this study, three chicken vaccination trials with different regimens were performed to determine conditions for efficient production of anti-Ent egg yolk IgY. Purified Ent was conjugated to three carrier proteins, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA) and CmeC (a subunit vaccine candidate), respectively. Intramuscular immunization of Barred Rock layers with KLH-Ent conjugate four times induced strong immune response against whole conjugate vaccine but the titer of Ent-specific IgY did not change in yolk with only a 4 fold increase detected in serum. In the second trial, three different Ent conjugate vaccines were evaluated in Rhode Island Red pullets with four subcutaneous injections. The KLH-Ent or CmeC-Ent conjugate consistently induced high level of Ent-specific IgY in both serum (up to 2,048 fold) and yolk (up to 1,024 fold) in each individual chicken. However, the Ent-specific immune response was only temporarily and moderately induced using a BSA-Ent vaccination. In the third trial, ten White Leghorn layers were subcutaneously immunized three times with KLH-Ent, leading to consistent and strong immune response against both whole conjugate and the Ent molecule in each chicken; the mean titer of Ent-specific IgY increased approximately 32 and 256 fold in serum and yolk, respectively. Consistent with its potent binding to various Ent derivatives, the Ent-specific egg yolk IgY also inhibited in vitro growth of a representative Escherichia coli strain. Together, this study demonstrated that the novel Ent conjugate vaccine could induce strong, specific, and robust immune response in chickens. The Ent-specific hyperimmune egg yolk IgY has potential for passive immune intervention against Gram-negative infections.

Characterizing avian pathogenic Escherichia coli (APEC) from colibacillosis cases, 2018.

Colibacillosis caused by avian pathogenic Escherichia coli (APEC) is a devastating disease of poultry that results in multi-million-dollar losses annually to the poultry industry. Disease syndromes associated with APEC includes colisepticemia, cellulitis, air sac disease, peritonitis, salpingitis, omphalitis, and osteomyelitis among others. A total of 61 APEC isolates collected during the Fall of 2018 (Aug-Dec) from submitted diagnostic cases of poultry diagnosed with colibacillosis were assessed for the presence of 44 virulence-associated genes, 24 antimicrobial resistance genes and 17 plasmid replicon types. Each isolate was also screened for its ability to form biofilm using the crystal violet assay and antimicrobial susceptibility to 14 antimicrobials using the NARMS panel. Overall, the prevalence of virulence genes ranged from 1.6% to >90% with almost all strains harboring genes that are associated with the ColV plasmid-the defining trait of the APEC pathotype. Overall, 58 strains were able to form biofilms and only three strains formed negligible biofilms. Forty isolates displayed resistance to antimicrobials of the NARMS panel ranging from one to nine agents. This study highlights that current APEC causing disease in poultry possess virulence and resistance traits and form biofilms which could potentially lead to challenges in colibacillosis control.

Citrate utilization under anaerobic environment in Escherichia coli is under direct control of Fnr and indirect control of ArcA and Fnr via CitA-CitB system.

Most Escherichia coli (E. coli) strains do not cause disease, naturally living in the lower intestine and is expelled into the environment within faecal matter. Escherichia coli can utilize citrate under anaerobic conditions but not aerobic conditions. However, the underlying regulatory mechanisms are poorly understood. In this study, we explored regulatory mechanisms of citrate fermentation genes by global regulators ArcA and Fnr under anaerobic conditions. A gel mobility shift assay showed that the regulator proteins ArcA and Fnr binded to the promoter region localized between the citAB and citCDEFXGT operons. Subsequent assays confirmed that ArcA indirectly controled the expression of citrate fermentation genes via regulating CitA-CitB system, while Fnr directly regulated but also indirectly modulated citrate fermentation genes via controling CitA-CitB system. Deletions of arcA and fnr significantly reduced the growth of Escherichia coli in M9 medium with a citrate carbon source. We conclude that both ArcA and Fnr can indirectly control the citrate utilization via CitA-CitB system, while Fnr can also directly regulate the expression of citrate fermentation genes in E. coli under anaerobic conditions.

Motivational Interviewing Training for HIV Care Physicians in Argentina: Uptake and Sustainability of an Effective Behavior Change Intervention.

Motivational interviewing (MI) utilizes a patient-centered approach to address patient ambivalence about treatment and has been found to improve treatment ART adherence among patients living with HIV disengaged from care. This study examined MI training for clinicians, uptake, and sustainability over time. Clinics (n = 7) with N = 38 physicians were randomized to condition (MI, Enhanced Standard of Care). Physicians completed video- recorded patient consultations at baseline and 6, 12, and 18-month follow-up. MI condition physicians had greater relational and technical scores over time and were more likely to adhere to and sustain MI over time. Overall, physicians found the MI training highly acceptable and were able to sustain their skills. Results illustrate the feasibility of MI training, implementation, and sustainment over 18 months. Findings support previous research in Argentina in which trained physicians found MI useful with challenging patients. Broader implementation of MI among HIV care physicians in Argentina is merited.

RESUMEN: La entrevista motivacional (EM) es una intervención con enfoque en el paciente que sirve para abordar la ambivalencia del paciente sobre el tratamiento. Se ha descubierto que EM mejora la adherencia al tratamiento antirretroviral en pacientes con VIH y que no están recibiendo tratamiento. En este estudio se examinó la capacitación de EM entre médicos, como también la aceptación y la sostenibilidad de EM a lo largo del tiempo. Las clínicas (n = 7) con N = 38 médicos se asignaron al azar a la condición (EM o estándar de atención mejorada). Los médicos hicieron consultas de pacientes, las cuales fueron grabadas en video al inicio del estudio y a los 6, 12 y 18 meses de seguimiento. Los médicos de la condición de EM obtuvieron puntuaciones relacionales y técnicas más altas a lo largo del tiempo y fueron más propensos ha adherirse y usar la EM con el tiempo. En general, los médicos consideraron que la formación en EM era muy aceptable y pudieron mantener sus habilidades. Los resultados ilustran la viabilidad de la capacitación, implementación y mantenimiento de la EM durante 18 meses. Los descubrimientos sustentan investigaciones anteriores en Argentina en las que médicos capacitados encontraron que la EM era útil para pacientes desafiantes. Una implementación más amplia de la EM entre los médicos de atención del VIH en Argentina es necesaria para mejorar el tratamiento de personas con VIH.

Characterizing the Type 6 Secretion System (T6SS) and its role in the virulence of avian pathogenic Escherichia coli strain APECO18.

Avian pathogenic E. coli is the causative agent of extra-intestinal infections in birds known as colibacillosis, which can manifest as localized or systemic infections. The disease affects all stages of poultry production, resulting in economic losses that occur due to morbidity, carcass condemnation and increased mortality of the birds. APEC strains have a diverse virulence trait repertoire, which includes virulence factors involved in adherence to and invasion of the host cells, serum resistance factors, and toxins. However, the pathogenesis of APEC infections remains to be fully elucidated. The Type 6 secretion (T6SS) system has recently gained attention due to its role in the infection process and protection of bacteria from host defenses in human and animal pathogens. Previous work has shown that T6SS components are involved in the adherence to and invasion of host cells, as well as in the formation of biofilm, and intramacrophage bacterial replication. Here, we analyzed the frequency of T6SS genes hcp, impK, evpB, vasK and icmF in a collection of APEC strains and their potential role in virulence-associated phenotypes of APECO18. The T6SS genes were found to be significantly more prevalent in APEC than in fecal E. coli isolates from healthy birds. Expression of T6SS genes was analyzed in culture media and upon contact with host cells. Mutants were generated for hcp, impK, evpB, and icmF and characterized for their impact on virulence-associated phenotypes, including adherence to and invasion of host model cells, and resistance to predation by Dictyostelium discoideum. Deletion of the aforementioned genes did not significantly affect adherence and invasion capabilities of APECO18. Deletion of hcp reduced resistance of APECO18 to predation by D. discoideum, suggesting that T6SS is involved in the virulence of APECO18.

Effects of Sugar-Sweetened, Artificially Sweetened, and Unsweetened Beverages on Cardiometabolic Risk Factors, Body Composition, and Sweet Taste Preference: A Randomized Controlled Trial.

Background A 2018 American Heart Association science advisory indicated that, pending further research, artificially sweetened beverages (ASBs) may be an appropriate initial replacement for sugar-sweetened beverages (SSBs) during transition to unsweetened beverages (USBs). Methods and Results We randomly assigned 203 adults (121 males, 82 females; 91.6% retention), who habitually consumed SSBs, to 3 groups and delivered free SSBs, ASBs, or USBs to their homes for 12 months. Outcomes included serum triglyceride to high-density lipoprotein cholesterol ratio (primary), body weight, and sweet taste preference (experimental assessment, 0%-18% sucrose solutions). Change in serum triglyceride to high-density lipoprotein cholesterol ratio was not different between groups. Although overall change in weight also was not different between groups, we found effect modification (P=0.006) by central adiposity. Among participants in the highest tertile of baseline trunk fat but not other tertiles, weight gain was greater (P=0.002) for the SSB (4.4±1.0 kg, estimate±SE) compared with ASB (0.5±0.9 kg) or USB (-0.2±0.9 kg) group. Both sweetness threshold (-1.0±0.2% m/v; P=0.005) and favorite concentration (-2.3±0.4% m/v; P<0.0001) decreased in the USB group; neither changed in the SSB group. In the ASB group, sweetness threshold did not change, and favorite concentration decreased (-1.1±0.5% m/v; P=0.02). Pairwise comparison between the ASB and USB groups indicated a difference in sweetness threshold (P=0.015). Conclusions Replacing SSBs with noncaloric beverages for 12 months did not affect serum triglyceride to high-density lipoprotein cholesterol ratio. Among individuals with central adiposity, replacing SSBs with either ASBs or USBs lowered body weight. However, USBs may have the most favorable effect on sweet taste preference. Registration URL: https://www.clinicaltrials.gov; unique identifier: NCT01295671.

Characterization of Avian Pathogenic Escherichia coli (APEC) Associated With Turkey Cellulitis in Iowa.

Turkey cellulitis, also known as clostridial dermatitis is a significant cause of morbidity, mortality, and carcass condemnation at slaughter resulting in considerable losses for turkey producers. Here, we assessed the potential role of Avian Pathogenic Escherichia coli (APEC) in a cellulitis outbreak on a turkey farm in Iowa. Birds from one farm with a history of cellulitis and one farm with no history of disease (for comparison) were followed from the age of 10 weeks (before the outbreak) to 18 weeks (just prior to slaughter). E. coli recovered from the litter, from skin lesions of birds with cellulitis, and from systemic lesions of birds submitted for necropsy, were assessed. A total of 333 isolates were analyzed and screened for virulence-associated genes, antimicrobial resistance genes including heavy metal resistance, adhesins, invasins, and protectins, iron acquisition systems and their phylogenetic group through multiplex PCR. In addition, PCR was used to serogroup the isolates, and pulsed field gel electrophoresis (PFGE) was used to analyze a subset of strains from the farm environment (litter) and birds at 17 and 18 weeks of age when the cellulitis infection appeared to peak. Overall, E. coli isolates recovered from cellulitis lesions and systemic infection were identified as APEC, while a lower prevalence of E. coli recovered from the litter met the criteria of APEC-like. Direct comparison of E. coli isolates from the litter, lesions, and systemic strains using PFGE failed to find identical clones across all three sources reflecting the diversity of strains present in the poultry environment causing disease. This study highlights the role of APEC in turkey cellulitis and should not be overlooked as a significant contributor to the disease in turkeys.

Outer membrane protein A (OmpA) of extraintestinal pathogenic Escherichia coli.

OBJECTIVE: Extraintestinal Pathogenic E. coli (ExPEC), are responsible for host diseases such as Neonatal Meningitis Escherichia coli (NMEC), the second-leading cause of neonatal bacterial meningitis, Avian Pathogenic E. coli (APEC), a cause of extraintestinal disease in poultry, and Uropathogenic E. coli (UPEC), the most common cause of urinary tract infections. Virulence factors associated with NMEC include outer membrane protein A (OmpA) and type I fimbriae (FimH), which also occur in APEC and UPEC. OmpA contributes to NMEC's ability to cross the blood-brain barrier, persist in the bloodstream and has been identified as a potential vaccine target for ExPEC, however the protein has amino acid variants, which may influence virulence of strains or alter vaccine efficacy. Although OmpA is present in virtually all E. coli, differences in its amino acid residues have yet to be surveyed in ExPEC. RESULTS: Here the ompA gene (n = 399) from ExPEC collections were sequenced and translated in silico. Twenty-five different OmpA polymorphism patterns were identified. Seven polymorphism patterns were significantly associated with an ExPEC subpathotype, but chromosomal history most likely accounts for most differences found. The differences in OmpA protein sequences suggest that OmpA may influence variation in virulence and host specificity within ExPEC subpathotypes.

Accuracy of PCR universal primer for methicillin-resistant Staphylococcus and comparison of different phenotypic screening assays.

Characterization of methicillin-resistant Staphylococcus (MRS) is a continuous challenge at diagnostic laboratories. The phenotypic methods present heterogeneous results and the occurrence of variants of mecA gene turned this goal even more challenging to achieve. The present study provided an accurate and highly discriminatory screening tool for MRS, improving its detection.

Assessing the Ability of Maternal Antibodies to Protect Broiler Chicks Against Colonization by Salmonella Heidelberg.

Vaccine regimes and maternal antibody protection are important in ensuring the health of poultry and critically important in decreasing contamination of poultry products with foodborne pathogens that threaten human health. Here, we assessed the role of passive immunity on the resistance of progeny to early colonization when challenged with Salmonella Heidelberg. Two broiler breeder hen flocks which had received a Salmonella vaccine regime consisting of two live attenuated and two killed vaccines during rearing were selected for study. ELISA titers were used to assess antibody levels in the parent flocks, with one low- and one high- titer flock selected for study. Progeny chicks (1 day of age) were taken from each flock and challenged with Salmonella Heidelberg at low (103) or high doses (105) at 3 days of age. At 14 days postinoculation, all birds were euthanatized and their liver, spleen, and ceca collected for culture. ELISA analysis found Flock A (30-wk flock) demonstrated higher Salmonella antibody titers in the parents as well as yolk titers in the progeny, resulting in greater early protection from colonization by Salmonella Heidelberg while Salmonella colonization rates were higher in the progeny of the older parent Flock (B) that demonstrated lower antibody titers in the parents and the yolks of the progeny. These results suggest that as the breeder hens' protective antibodies wane with age; the maternal antibody protection in their progeny also becomes less effective in preventing cecal colonization by Salmonella early in life, which has the potential for affecting the health of the bird and contamination of meat products destined for the consumer.

Evaluación de la capacidad de los anticuerpos maternos para proteger a pollos de engorde contra la colonización por Salmonella Heidelberg. Los esquemas de vacunas y la protección por anticuerpos maternos son importantes para garantizar la salud de las aves comerciales y críticamente importantes para reducir la contaminación de los productos avícolas con patógenos transmitidos a través de los alimentos que amenazan la salud humana. En este estudio, se evaluó el papel de la inmunidad pasiva en la resistencia de la progenie contra la colonización temprana cuando se la desafió con Salmonella Heidelberg. Dos parvadas de gallinas reproductoras de pollos de engorde que habían recibido un esquema de vacuna de Salmonella que consistía en dos vacunas vivas atenuadas y dos muertas durante la crianza se seleccionaron para el estudio. Los títulos de ELISA se utilizaron para evaluar los niveles de anticuerpos en las parvadas parentales y se seleccionó para el estudio una parvada con títulos bajos y una con títulos altos. La progenie (pollos de un día de edad) de cada parvada se recolectó y se desafiaron con Salmonella Heidelberg a dosis bajas (103) y altas (105), a los tres días de edad. A los 14 días posteriores a la inoculación, todas las aves se sometieron a eutanasia y se recolectaron hígado, bazo y ciego para el cultivo. El análisis por ELISA demostró que la parvada A (lote de 30 semanas) mostraba mayores títulos de anticuerpos de Salmonella en las reproductoras, así como títulos en el saco vitelino de la progenie, lo que resultó en una mayor protección temprana contra la colonización por Salmonella Heidelberg, mientras que las tasas de colonización de Salmonella fueron mayores en la progenie de la parvada de reproductoras (B) que demostró títulos de anticuerpos más bajos en las reproductoras y en el saco vitelino de la progenie. Estos resultados sugieren que a medida que los anticuerpos protectores de las gallinas reproductoras disminuyen con la edad, la protección de los anticuerpos maternos en su progenie también se vuelve menos efectiva en la prevención de la colonización cecal por Salmonella en edades tempranas, lo que tiene el potencial de afectar la salud de las aves y la contaminación de los productos cárnicos destinados al consumidor.

Complete Genome Sequence of the Multidrug-Resistant Neonatal Meningitis Escherichia coli Serotype O75:H5:K1 Strain mcjchv-1 (NMEC-O75).

Neonatal meningitis Escherichia coli (NMEC) is the second leading cause of neonatal bacterial meningitis worldwide. We report the genome sequence of the multidrug-resistant NMEC serotype O75:H5:K1 strain mcjchv-1, which resulted in an infant's death. The O75 serogroup is rare among NMEC isolates; therefore, this strain is considered an emergent pathogen.

Complete Genome Sequence of Avian Pathogenic Escherichia coli Strain APEC O2-211.

Avian pathogenic Escherichia coli (APEC) is the causative agent of colibacillosis, a disease that affects poultry production worldwide and leads to multimillion-dollar losses annually. Here, we report the genome sequence of APEC O2-211, a sequence type 117 (ST117) strain isolated from a diseased chicken.