Molekulare Aspekte und chemische Inaktivierung von Influenza H5N1-Viren aus ägyptischen Hühnerbeständen von Ausbrüchen der Jahre 2006 bis 2010

The primary objective of the current study was to identify two of the highly pathogenic avian influenza virus (HPAIV) isolates of subtype H5N1 genotypically using one step Reverse Transcriptase Polymerase Chain Reaction (RT-PCR), followed by sequence and phylogenetic analyses. A further objective was to determine in vitro the virucidal efficacy of four types of chemical disinfectants, namely Formalin, Glutaraldehyde, TH4® and Virkon®S at different concentrations and contact times on the two HPAI isolates. A/chicken/Egypt/0626/2006 (EGY06) and A/chicken/Egypt/1094/2010 (EGY10) were isolated from cloacal and tracheal swabs from broiler during HPAI H5N1 outbreaks in Egypt in 2006 and 2010. The first strain, EGY06, was isolated from a non-vaccinated flock in February 2006 in the Alexandria governorate. The second strain, EGY10, was isolated from a vaccinated flock in November 2010 in the Marsa Matrouh governorate. Classical identification of the two isolates was carried out in the Department of Poultry and Hygiene, Faculty of Veterinary Medicine, Alexandria University, Egypt. Molecular identification and genetic analyses were conducted in the Gene Analysis Unit of the National Laboratory for Veterinary Quality Control on Poultry Production (NLQP), Egypt. Using RT-PCR with specific sets of primers for H5 and N1 genes of AIV it was confirmed that the two isolates belonged to AI subtype H5N1. After molecular characterization and phylogenetic analysis of the HA and NA genes, the strain EGY06 was closely related to the 2006 predecessor Egyptian viruses of 2.2.1 clade, whereas EGY10 clustered within the classic 2.2.1/c group that commonly isolated from small-scale commercial farms and human since 2009. The efficacy of four chemical disinfectants to inactivate both isolates was carried out in accordance to the guidelines of the German Veterinary Medical Society (Deutsche Veterinärmedizinische Gesellschaft, DVG) for testing of disinfection procedures and chemical disinfectants. The experiments were performed using suspension tests without and with protein load (40% Bovine Calf Serum "BCS") as well as wood and gauze as a carriers (also loaded with BCS), at room temperature and incubation times of 15 to 120 min. The obtained results showed that the use of Glutaraldehyde, Formalin or TH4® 0.5% without protein load led to complete inactivation of the virus after 15, 30, 60 or 120 min contact time. Use of Virkon®S 0.5% with and without protein load led to survival of the virus even after 60 min. In contrast, using Formalin and TH4® (1% and 2%) with and without protein load led to complete inactivation of the virus even at the shortest contact time of 15 min. Similar results were obtained after using Glutaraldehyde 1%, while treatment of H5N1 with Glutaraldehyde 2% led to gel formation. After treatment of contaminated carriers (poplar wood and gauze) with Formalin, Glutaraldehyde and TH4® 0.5%, the virus was inactivated after 30 min. Concentration of 1% of the three disinfectants was sufficient to inactivate the two isolates within 15 min contact time, except in case of Virkon®S which required higher concentrations to give similar results. The study indicated that the four chemical disinfectants could efficiently inactivate the two tested H5N1 viruses when used at higher concentration than the manufacturers recommended. The results of the present thesis highlight the sensitivity of HPAIV H5N1 to different disinfectants, which may improve biosecurity measures on the farms and reduce the economic losses caused by HPAIV H5N1.Das primäre Ziel der aktuellen Studie war es, hoch pathogene aviäre Influenza- Viren (HPAIV) des Subtyps H5N1 genotypisch durch eine einschrittige Reverse Transkriptase- Polymerase-Kettenreaktion (RT-PCR) zu identifizieren und anschließend molekularbiologisch zu charakterisieren. Ein weiteres Ziel war, die Wirksamkeit von verschiedenen Konzentrationen und Einwirkungszeiten von vier chemischen Desinfektionsmitteln (Formalin, Glutaraldehyd, TH4® und Virkon®S) auf zwei Stämme (A/chicken/Egypt/0626/2006 "EGY06" und A/chicken/Egypt/1094/2010 "EGY10") des aviären Influenzavirus (AIV) des Subtyps H5N1 in vitro zu prüfen. Die beiden Isolate des AIV-Subtyps H5N1 wurden aus Kloaken- und Trachealtupfern von infizierten Masthühnerherden während der Ausbrüche aviärer Influenza (AI) 2006 und 2010 isoliert. Während der erste Stamm EGY06 aus einer nicht geimpften Herde im Februar 2006 im Gouvernement Alexandria isoliert wurde, wurde der zweite Stamm, EGY10, aus einer geimpften Herde im November 2010 im Gouvernement Marsa Matrouh isoliert. Die klassischen Methoden zur Identifizierung der beiden Isolate wurden in der Abteilung für Geflügel und Hygiene, Veterinärmedizinische Fakultät, Universität Alexandria, Ägypten durchgeführt. Die molekulare Identifizierung und genetische Analyse erfolgten in der Gen- Analyse-Einheit des Nationalen Labors zur Qualitätskontrolle der Geflügelproduktion (NLQP), Ägypten. Mittels RT-PCR unter Verwendung spezifischer Primersets für die H5 und N1 Gene konnte bestätigt werden, dass es sich bei beiden Isolaten um AIV des Subtyps H5N1 handelt. Der molekularen Charakterisierung und der phylogenetischen Analyse der HA und NA zufolge war der Stamm EGY06 sehr eng verwandt mit dem früher im Jahr 2006 isolierten klassischen Stamm und wurde dem Clade 2.2.1 zugeordnet. Im Gegensatz dazu wurde der Stamm EGY10 im klassischen 2.2.1/c Gruppe zugeordnet, welcher häufig von kleinen kommerziellen Farmen und menschlichen seit 2009 isoliert. Die Empfindlichkeit der Viren gegen verschiedene Desinfektionsmittel wurde auf Grundlage der Richtlinien der Deutschen Veterinärmedizinischen Gesellschaft (DVG) für die Prüfung von Desinfektionsverfahren und chemischen Desinfektionsmitteln geprüft. Die Experimente wurden mittels Suspensions-Test ohne und mit Proteinbelastung (40% Bovines Calf Serum (BCS)) sowie auf Keimträgern aus Holz und Gaze, die mit BCS belastet waren, bei Raumtemperatur und Einwirkzeiten von 15 bis 120 Min durchgeführt. Die Verwendung von Glutaraldehyd, Formalin oder TH4® in einer Konzentration von 0,5% führte ohne Proteinbelastung zu einer Inaktivierung der Viren nach allen Einwirkzeiten (15, 30, 60 und 120 Min). Die Verwendung von Virkon®S 0,5% mit und ohne Proteinbelastung führte zum Überleben des Virus sogar nach 60 Min. Demgegenüber führte die Verwendung von Formalin und TH4® in einer Konzentration von 1% und 2% mit und ohne Proteinbelastung zu einer vollständigen Inaktivierung des Virus sogar bei der kürzesten Einwirkungszeit von 15 Min. Ähnliche Ergebnisse wurden nach Verwendung von Glutaraldehyd in einer Konzentration von 1% beobachtet. Die Behandlung von H5N1 mit Glutaraldehyd in einer Konzentration von 2% führte zu einer Gelbildung. Nach der Behandlung von kontaminierten Keimträgern (Pappelholz und Gaze) mit Formalin, Glutaraldehyd und TH4® in Konzentrationen von 0,5% wurde das Virus nach 30 Min inaktiviert. Während eine Konzentration von 1% der drei Desinfektionsmittel ausreichend war, um die beiden Isolate in 15 Min Einwirkzeit zu inaktivieren, konnte dieses Ergebnis im Fall von Virkon®S nicht erreicht werden, und eine höhere Konzentration war erforderlich um ähnliche Ergebnisse zu erzielen. Die Studie zeigte, dass die vier chemischen Desinfektionsmittel, wenn die verwendeten Konzentrationen höher als die vom Hersteller empfohlenen Konzentrationen sind, beide getesteten H5N1 Viren effektiv inaktivieren. Die Ergebnisse dieser Studie bieten einen neuen Ansatz zur Verbesserung der Biosicherheitsmaßnahmen in Geflügelbeständen und können zur Reduzierung der wirtschaftlichen Verluste beitragen

An overview of the public health challenges in diagnosing and controlling human foodborne pathogens

Pathogens found in food are believed to be the leading cause of foodborne illnesses; and they are considered a serious problem with global ramifications. During the last few decades, a lot of attention has been paid to determining the microorganisms that cause foodborne illnesses and developing new methods to identify them. Foodborne pathogen identification technologies have evolved rapidly over the last few decades, with the newer technologies focusing on immunoassays, genome-wide approaches, biosensors, and mass spectrometry as the primary methods of identification. Bacteriophages (phages), probiotics and prebiotics were known to have the ability to combat bacterial diseases since the turn of the 20th century. A primary focus of phage use was the development of medical therapies; however, its use quickly expanded to other applications in biotechnology and industry. A similar argument can be made with regards to the food safety industry, as diseases directly endanger the health of customers. Recently, a lot of attention has been paid to bacteriophages, probiotics and prebiotics most likely due to the exhaustion of traditional antibiotics. Reviewing a variety of current quick identification techniques is the purpose of this study. Using these techniques, we are able to quickly identify foodborne pathogenic bacteria, which forms the basis for future research advances. A review of recent studies on the use of phages, probiotics and prebiotics as a means of combating significant foodborne diseases is also presented. Furthermore, we discussed the advantages of using phages as well as the challenges they face, especially given their prevalent application in food safety

Burnout among surgeons before and during the SARS-CoV-2 pandemic: an international survey

Background: SARS-CoV-2 pandemic has had many significant impacts within the surgical realm, and surgeons have been obligated to reconsider almost every aspect of daily clinical practice. Methods: This is a cross-sectional study reported in compliance with the CHERRIES guidelines and conducted through an online platform from June 14th to July 15th, 2020. The primary outcome was the burden of burnout during the pandemic indicated by the validated Shirom-Melamed Burnout Measure. Results: Nine hundred fifty-four surgeons completed the survey. The median length of practice was 10 years; 78.2% included were male with a median age of 37 years old, 39.5% were consultants, 68.9% were general surgeons, and 55.7% were affiliated with an academic institution. Overall, there was a significant increase in the mean burnout score during the pandemic; longer years of practice and older age were significantly associated with less burnout. There were significant reductions in the median number of outpatient visits, operated cases, on-call hours, emergency visits, and research work, so, 48.2% of respondents felt that the training resources were insufficient. The majority (81.3%) of respondents reported that their hospitals were included in the management of COVID-19, 66.5% felt their roles had been minimized; 41% were asked to assist in non-surgical medical practices, and 37.6% of respondents were included in COVID-19 management. Conclusions: There was a significant burnout among trainees. Almost all aspects of clinical and research activities were affected with a significant reduction in the volume of research, outpatient clinic visits, surgical procedures, on-call hours, and emergency cases hindering the training. Trial registration: The study was registered on clicaltrials.gov "NCT04433286" on 16/06/2020

Evaluation of Imaging-Guided Peritoneal Biopsy in Diagnosis of Ascites of Undetermined Origin

Abstract Background Regarding ascites of unknown origin, diagnostic laparoscopy is an invasive procedure, there are certain complications reported with this procedure i.e. haemorrhage, infection and air embolism. Ultrasound-guided percutaneous biopsies are easy to perform in an outpatient clinic. This procedure is safe, has a low incidence of injury and does not cause serious complications. Computed tomography (CT)-guided percutaneous biopsy is not a real-time operation, and it involves quite a few complicated procedures. Aim To evaluate the role of imaging guided peritoneal biopsy in diagnosis of ascites of undetermined origin (ascites of local cause). 2ry aim was to present the role of imaging-guided biopsy of the omentum or other extravisceral masses as a minimally invasive procedure compared to laparoscopy in the diagnosis of these difficult-to-diagnose group of patients. Patients and methods Patients with clinically suspected and radiologically confirmed ascites of unknown etiology represented the population of our study. These patients were referred to the ascites study group (ASG) and admitted to Tropical Medicine Department, Ain Shams University Hospitals in the period from June 2017 to November 2019. The study was conducted on 63 patients with ascites of unknown etiology fulfilling the inclusion criteria. They underwent ultrasound-guided cytology/biopsy of peritoneum, omentum or extravisceral masses. CT guided percutaneous peritoneal biopsy was done in cases of failure of ultrasound guided technique. Laparoscopy was needed when the imaging-guided biopsy was not diagnostic. Results 54 patients (85.7%) underwent US guided biopsies, 48 patients of them (76.2%) were successfully diagnosed, while the other six patients (9.5%) were sent for laparoscopy after nonconclusive histopathological examination of the biopsies taken US guided. The patients underwent laparoscopy were successfully diagnosed except for one patient who died intraoperatively. The other nine patients (14.3%) underwent CT guided biopsies (not accessible by US guided modality) and all of them were successfully diagnosed. Imaging guided biopsies had perfect sensitivity (100%) and NPV (100%) in differentiating neoplastic lesions. We found that imaging-guided procedures had a high diagnostic accuracy of 88.8% &amp; 100% done US &amp; CT guided respectively with a sensitivity of 100%, specificity of 83.3% and NPV of 100%, which could distinguish malignant from benign ascites. Complications were most frequent in laparocopy, followed by CT guided biopsies and least in US guided biopsies with P-value &amp;lt;0.001. Conclusion Percutaneous imaging-guided biopsy (US/CT guided) of the peritoneum, omenta, and mesentery has been established as a safe, well-tolerated procedure with high diagnostic accuracy. It can minimize further unnecessary invasive procedures e.g laparoscopy. It can help in directing the management, shortening the patient’s hospital stay and reducing the costs and complications. </jats:sec

Pseudomonas Species Prevalence, Protein Analysis, and Antibiotic Resistance: An Evolving Public Health Challenge

Abstract Psychrotrophic Pseudomonas is one of the significant microbes that lead to putrefaction in chilled meat. One of the biggest problems in the detection of Pseudomonas is that several species are seemingly identical. Antibiotic resistance is an alternative considerable challenge worldwide. Therefore, this study was designed to apply an accurate technique for the discrepancy of Pseudomonas species and to study their resistance against various antimicrobials. A total of 320 chicken meat specimens were cultivated, and phenotypically recognized the isolated bacteria. Protein analysis was carried out for cultured isolates via Microflex LT. The resistance of Pseudomonas isolates was recorded through Vitek®ฏ 2 AST-GN83 cards. Overall, 69 samples were identified as Pseudomonas spp. and included 18 Pseudomonas lundensis (P. lundensis), 16 Pseudomonas fragi (P. fragi), 13 Pseudomonas oryzihabitans (P. oryzihabitans), 10 Pseudomonas stutzeri (P. stutzeri), 5 Pseudomonas fluorescens (P. fluorescens), 4 Pseudomonas putida (P. putida), and 3 Pseudomonas aeruginosa (P. aeruginosa) isolates. Microflex LT identified all Pseudomonas isolates (100%) correctly with a score value ≥2.00. PCA positively discriminated the identified isolates into various groups. The antimicrobial resistance levels against Pseudomonas isolates were 81.16% for nitrofurantoin, 71% for ampicillin and ampicillin/sulbactam, 65.22% for cefuroxime and ceftriaxone, 55% for aztreonam, and 49.28% for ciprofloxacin. The susceptibilities were 100% for cefotaxime, 98.55% for ceftazidime, 94.20% for each piperacillin/tazobactam and cefepime, 91.3% for cefazolin. In conclusion, chicken meat was found to be contaminated with different Pseudomonas spp., with high incidence rates of P. lundensis. Microflex LT is a potent tool for distinguishing Pseudomonads at the species level.</jats:p

The Development of Diagnostic and Vaccine Strategies for Early Detection and Control of Human Brucellosis, Particularly in Endemic Areas

Brucellosis is considered one of the most serious zoonotic diseases worldwide. This disease affects both human and animal health, in addition to being one of the most widespread zoonotic illnesses in the Middle East and Northern Africa. Human brucellosis generally presents in a diverse and non-specific manner, making laboratory confirmation of the diagnosis critical to the patient’s recovery. A coordinated strategy for diagnosing and controlling brucellosis throughout the Middle East is required, as this disease cannot be known to occur without reliable microbiological, molecular, and epidemiological evidence. Consequently, the current review focuses on the current and emerging microbiological diagnostic tools for the early detection and control of human brucellosis. Laboratory assays such as culturing, serology, and molecular analysis can frequently be used to diagnose brucellosis. Although serological markers and nucleic acid amplification techniques are extremely sensitive, and extensive experience has been gained with these techniques in the laboratory diagnosis of brucellosis, a culture is still considered to be the “gold standard” due to the importance of this aspect of public health and clinical care. In endemic regions, however, serological tests remain the primary method of diagnosis due to their low cost, user-friendliness, and strong ability to provide a negative prediction, so they are commonly used. A nucleic acid amplification assay, which is highly sensitive, specific, and safe, is capable of enabling rapid disease diagnosis. Patients who have reportedly fully healed may continue to have positive molecular test results for a long time. Therefore, cultures and serological methods will continue to be the main tools for diagnosing and following up on human brucellosis for as long as no commercial tests or studies demonstrate adequate interlaboratory reproducibility. As there is no approved vaccine that prevents human brucellosis, vaccination-based control of animal brucellosis has become an important part of the management of human brucellosis. Over the past few decades, several studies have been conducted to develop Brucella vaccines, but the problem of controlling brucellosis in both humans and animals remains challenging. Therefore, this review also aims to present an updated overview of the different types of brucellosis vaccines that are currently available.</jats:p

The Development of Diagnostic and Vaccine Strategies for Early Detection and Control of Human Brucellosis, Particularly in Endemic Areas

Brucellosis is considered one of the most serious zoonotic diseases worldwide. This disease affects both human and animal health, in addition to being one of the most widespread zoonotic illnesses in the Middle East and Northern Africa. Human brucellosis generally presents in a diverse and non-specific manner, making laboratory confirmation of the diagnosis critical to the patient&rsquo;s recovery. A coordinated strategy for diagnosing and controlling brucellosis throughout the Middle East is required, as this disease cannot be known to occur without reliable microbiological, molecular, and epidemiological evidence. Consequently, the current review focuses on the current and emerging microbiological diagnostic tools for the early detection and control of human brucellosis. Laboratory assays such as culturing, serology, and molecular analysis can frequently be used to diagnose brucellosis. Although serological markers and nucleic acid amplification techniques are extremely sensitive, and extensive experience has been gained with these techniques in the laboratory diagnosis of brucellosis, a culture is still considered to be the &ldquo;gold standard&rdquo; due to the importance of this aspect of public health and clinical care. In endemic regions, however, serological tests remain the primary method of diagnosis due to their low cost, user-friendliness, and strong ability to provide a negative prediction, so they are commonly used. A nucleic acid amplification assay, which is highly sensitive, specific, and safe, is capable of enabling rapid disease diagnosis. Patients who have reportedly fully healed may continue to have positive molecular test results for a long time. Therefore, cultures and serological methods will continue to be the main tools for diagnosing and following up on human brucellosis for as long as no commercial tests or studies demonstrate adequate interlaboratory reproducibility. As there is no approved vaccine that prevents human brucellosis, vaccination-based control of animal brucellosis has become an important part of the management of human brucellosis. Over the past few decades, several studies have been conducted to develop Brucella vaccines, but the problem of controlling brucellosis in both humans and animals remains challenging. Therefore, this review also aims to present an updated overview of the different types of brucellosis vaccines that are currently available

Emerging Technologies and Integrated Strategies for Microbial Detection and Control in Fresh Produce

The global consumption of fresh and ready-to-eat (RTE) fruits and vegetables has surged due to increasing awareness of their nutritional benefits. However, this trend has been accompanied by a rise in foodborne illness outbreaks linked to microbial contamination. This narrative review synthesizes current knowledge on the prevalence and diversity of foodborne pathogens in fresh produce, including bacterial, viral, and fungal agents. It critically evaluates both conventional and emerging detection methods, ranging from culture-based techniques and immunoassays to advanced molecular diagnostics, biosensors, flow cytometry (FC), and hyperspectral imaging (HSI). Additionally, this review discusses cutting-edge control strategies, such as natural antifungal agents, essential oils, biocontrol methods, and non-thermal technologies like cold plasma and UV-C treatment. Emphasis is placed on sampling methodologies, sustainability, One Health perspectives, and regulatory considerations. By highlighting recent technological advances and their limitations, this review aims to support the development of integrated, effective, and safe microbial control approaches for the fresh produce supply chain

Perceived clinical competence and its related factors among registered nurses employed at selected outpatient clinics in Egypt.

Nurses' clinical competence is a significant concern in all healthcare settings due to the necessity of delivering high-quality patient care. Understanding and addressing the factors related to competence are crucial for promoting nurses' clinical competence and ultimately improving patient outcomes. Producing and maintaining a skilled nursing workforce is essential to protect communities. This study aimed to assess the level of self-evaluated clinical competence and its correlation with demographic and occupational variables among registered nurses employed at selected outpatient clinics in Egypt. The study utilized a descriptive cross-sectional design with a self-administered, two-part questionnaire that assessed participants' demographic and occupational variables as well as perceived clinical competence in various healthcare settings. It took place at outpatient clinics of two governmental hospitals and five primary healthcare centers in Mansoura City, Egypt between January, and June 2023. A purposive sample of 450 nurses took part in this study. The average score of nurses' clinical competence was 155.3±7.2 out of 230, indicating a "moderate level". In terms of professional behaviors and general performance, the average score for clinical competence was 48.4±3.6 and 40.7±4.1 respectively. Additionally, the average score for clinical competence regarding core and advanced nursing skills were 43.4±3.0 and 22.8±1.5 respectively. Among the domains of clinical competence, the highest average score was associated with "professional behaviors" as it forms the backbone of nursing practice. There was a highly significant relationship between the average score of clinical competence and the participant's age, sex, level of education, and years of clinical work experience (P<0.001). Nurses perceived their level of clinical competence as moderate. To enhance nurses' clinical competence, future studies and interventions should focus on promoting supportive work environments, providing ongoing education and training in advanced nursing skills, and the fostering development of critical thinking skills in nurses. Healthcare organizations should implement educational interventions to enhance nurses' clinical competence. These interventions should include continuous professional development opportunities, mentorship programs, inclusive training initiatives, and structured feedback mechanisms. These measures will help nurses stay up-to-date with the latest practices and technologies, create a supportive learning atmosphere, and address the unique needs and challenges faced by nurses of different genders and specialties