Distribution pattern of antibiotic resistance genes in Escherichia coli isolated from colibacillosis cases in broiler farms of Egypt

Background and Aim: Multidrug resistance (MDR) of Escherichia coli has become an increasing concern in poultry farming worldwide. However, E. coli can accumulate resistance genes through gene transfer. The most problematic resistance mechanism in E. coli is the acquisition of genes encoding broad-spectrum β-lactamases, known as extended-spectrum β-lactamases, that confer resistance to broad-spectrum cephalosporins. Plasmid-mediated quinolone resistance genes (conferring resistance to quinolones) and mcr-1 genes (conferring resistance to colistin) also contribute to antimicrobial resistance. This study aimed to investigate the prevalence of antimicrobial susceptibility and to detect β-lactamase and colistin resistance genes of E. coli isolated from broiler farms in Egypt. Materials and Methods: Samples from 938 broiler farms were bacteriologically examined for E. coli isolation. The antimicrobial resistance profile was evaluated using disk diffusion, and several resistance genes were investigated through polymerase chain reaction amplification. Results: Escherichia coli was isolated and identified from 675/938 farms (72%) from the pooled internal organs (liver, heart, lung, spleen, and yolk) of broilers. Escherichia coli isolates from the most recent 3 years (2018–2020) were serotyped into 13 serotypes; the most prevalent serotype was O125 (n = 8). The highest phenotypic antibiotic resistance profiles during this period were against ampicillin, penicillin, tetracycline, and nalidixic acid. Escherichia coli was sensitive to clinically relevant antibiotics. Twenty-eight selected isolates from the most recent 3 years (2018–2020) were found to have MDR, where the prevalence of the antibiotic resistance genes ctx, tem, and shv was 46% and that of mcr-1 was 64%. Integrons were found in 93% of the isolates. Conclusion: The study showed a high prevalence of E. coli infection in broiler farms associated with MDR, which has a high public health significance because of its zoonotic relevance. These results strengthen the application of continuous surveillance programs

Impact of opioid-free analgesia on pain severity and patient satisfaction after discharge from surgery: multispecialty, prospective cohort study in 25 countries

Background: Balancing opioid stewardship and the need for adequate analgesia following discharge after surgery is challenging. This study aimed to compare the outcomes for patients discharged with opioid versus opioid-free analgesia after common surgical procedures.Methods: This international, multicentre, prospective cohort study collected data from patients undergoing common acute and elective general surgical, urological, gynaecological, and orthopaedic procedures. The primary outcomes were patient-reported time in severe pain measured on a numerical analogue scale from 0 to 100% and patient-reported satisfaction with pain relief during the first week following discharge. Data were collected by in-hospital chart review and patient telephone interview 1 week after discharge.Results: The study recruited 4273 patients from 144 centres in 25 countries; 1311 patients (30.7%) were prescribed opioid analgesia at discharge. Patients reported being in severe pain for 10 (i.q.r. 1-30)% of the first week after discharge and rated satisfaction with analgesia as 90 (i.q.r. 80-100) of 100. After adjustment for confounders, opioid analgesia on discharge was independently associated with increased pain severity (risk ratio 1.52, 95% c.i. 1.31 to 1.76; P < 0.001) and re-presentation to healthcare providers owing to side-effects of medication (OR 2.38, 95% c.i. 1.36 to 4.17; P = 0.004), but not with satisfaction with analgesia (beta coefficient 0.92, 95% c.i. -1.52 to 3.36; P = 0.468) compared with opioid-free analgesia. Although opioid prescribing varied greatly between high-income and low- and middle-income countries, patient-reported outcomes did not.Conclusion: Opioid analgesia prescription on surgical discharge is associated with a higher risk of re-presentation owing to side-effects of medication and increased patient-reported pain, but not with changes in patient-reported satisfaction. Opioid-free discharge analgesia should be adopted routinely

Detection of aerobic bacterial pathogens associated with early embryonic death in pregnant New Zealand female Rabbits in Egypt

Background and Aim: Rabbits are a highly sensitive species and susceptible to various bacterial pathogens that may be causative agents for early embryonic death. This study aimed to explore the administration of different bacterial agents in does suffering from early embryonic death. Furthermore, identification of genes associated with virulence was performed to identify the phenotypic and genotypic antimicrobial resistance patterns that may increase the virulence of pathogens and lead to early embryonic death. Materials and Methods: We isolated and identified bacterial agents in 106 samples from live and dead female rabbits that had undergone early embryonic death, including liver and intestine tissue, aborted fetuses, discharges, and vaginal swabs. Conventional polymerase chain reaction (PCR) was conducted to confirm the identity of the isolated bacterial strains and their virulence. Moreover, antibiotic resistance was studied phenotypically and genotypically. Results: We isolated Escherichia coli, Salmonella, Staphylococcus aureus, Pasteurella multocida, and Listeria monocytogenes. PCR confirmed typical identification except in P. multocida, which was confirmed as Gallibacterium spp. in some cases. The final percentage of detection was 34%, 30.2%, 16.9%, 13.2%, and 11.3%, respectively. Virulence properties were investigated using different designated genes. All Salmonella strains harbored invA, stn, avrA, and ompf genes, while the sopE gene was identified in 31.25%. E. coli strains harboring the iss gene lacked the shiga toxin (stx1) gene. L. monocytogenes and S. aureus strains harbored the hemolysin gene (66.7% and 33.4%, respectively). Multidrug resistance was detected phenotypically and genotypically in most strains. Each bacterial pathogen had a different antibiotic resistance profile. Conclusion: Multiple bacterial species may contribute to early embryonic death in does. Furthermore, the combined infection could be the main cause of early embryonic death. Thus, monitoring programs should bear this in mind and focus on the early detection of these bacterial agents in female rabbits to avoid embryonic death

Immunohistochemical Expression of HER2/neu, Ki-67 and MUC1 in Benign and Malignant Gall Bladder Lesions and its Association with Clinicopathological Parameters

Introduction: Gall Bladder Carcinoma (GBC) is a diagnostic and a therapeutic challenge. Although it is increasing, chronic cholecystitis remains the most worldwide gall bladder lesions, harbouring many epithelial changes that may end in carcinoma. Aim: To investigate the expression of HER2/neu (Human Epidermal Growth Factor Receptor 2), Ki-67 and MUC1 (Mucin 1) in malignant and non-malignant gall bladder lesions, and to evaluate its relation with clinicopathologic parameters of GBC. Materials and Methods: This retrospective study included 40 cases of GBC, eight cases of gall bladder dysplasia, 10 cases of gall bladder metaplastic changes and 25 cases of chronic cholecystitis as a control group. The blocks were collected from the Department of Pathology of Benha University Hospital, from January 2012 to December 2019. Immunohistochemical staining results of HER2/neu, Ki-67 and MUC1 were analysed and correlated by Statistical Package for the Social Sciences (SPSS) version 16 and Chi-square test or Fisher’s-exact tests. Results: Positive HER2/neu expression (+2, +3) was detected in 47.5% (19/40) of malignant cases and 12.5% (1/8) of dyspastic group, at the same time it was completely absent in the metaplastic and cholecystitis cases (p<0.01). Similarly, Ki-67 Labeling Index (LI) (≥20%) expression was found in 55% (22/40) of malignant group, while it was completely absent in the other three studied groups. All cases of malignant group 100% (40/40), 50% (4/8) of dysplastic one, one case of metaplastic (1/10) showed cytoplasmic expression of MUC1, at the same time it was completely absent in control group (0/25) (p<0.01). High MUC1 expression was found in 75% of both malignant (30/40) and dysplastic (6/8) studied cases and only one case (10%) of metaplastic group (p<0.01). There was a significant correlation between MUC1 expression and studied parameters of GBC. Conclusion: HER2/neu, and Ki-67 are overexpressed in GBC cases compared with control and dysplastic group. The study also highlights that MUC1 would be a better marker of malignant transformation of gall bladder epithelium and its depolarised expression would be reliable for detection of invasive carcinoma, so a new therapeutic agent can target these cell surface adhesion molecule (MUC1). HER2/neu can be considered as a candidate for targeted therapy in GBC treatment strategy

Isolation and Molecular Identification of Avibacterium paragallinarum in Suspected Cases of Poultry

Infectious Coryza (IC) is an infectious upper respiratory disease of chickens and birds, caused by Avibacterium paragallinarum (Av. Paragallinarum). It is characterized by facial swelling and nasal discharge. Here, the study aimed to identify the causative agent of infectious coryza from diseased cases using conventional and molecular methods, forty-one different samples (swollen heads and nasal discharges) from layers and broiler chickens were examined bacteriologically for isolation of Av. Paragallinarum.  The total isolation percentage was 9.75%, and the percentage was 66.7% from total layer samples. The samples underwent examination by polymerase chain reaction (PCR) using HPG-2. Three samples were confirmed by PCR with a percentage of 7.3% (3/41). Multiplex PCR was used for typing the tested strains using HMTp210 gene thus, all strains were of type B. sequence analysis showed a maximum identity percentage (100%) between the tested strains of Av. paragallinarum. In conclusion, typical signs for IC diseases were appeared and identified bacteriologically as Av. paragallinarum also three strains were only confirmed by PCR and identified type B strains. PCR technique is a reliable and sensitive test for confirmation

Green quality by design HPLC approach for the simultaneous determination of Bilastine and Montelukast

Abstract For the simultaneous estimation of two co-formulated antihistaminic drugs (Bilastine and Montelukast), a novel and eco-friendly reversed-phase HPLC approach with both diode array and fluorescence detection modes was designed. Rather than using the routine methodology, the Quality by Design (QbD) approach was adopted to speed up the method development and to test robustness of the method. To evaluate the effect of variable factors on chromatographic response, a full factorial design was used. The chromatographic separation was performed using isocratic elution on the C18 column. The mobile phase consists of 92% methanol, 6% acetonitrile, and 2% phosphate buffer with 0.1 (v/v) triethylamine adjusted to pH 3, it was pumped at a flow rate of 0.8 mL/min with an injection volume of 20 μL. The developed stability indicating HPLC approach was used to assess the stability of montelukast (MNT). It was subjected to a variety of stress conditions, including hydrolytic (acid–base), oxidative, thermal, and photolytic stress conditions. All of these conditions were found to have relevant degradation pathways. Under the described experimental conditions, MNT degradation followed pseudo-first-order kinetics. The kinetic parameters of its degradation (rate constant and t1/2) were calculated and a proposal for the degradation pathway was postulated

Antibiotic resistance and extended-spectrum β-lactamase in Escherichia coli isolates from imported 1-day-old chicks, ducklings, and turkey poults

Aim: Antimicrobial resistance is a global health threat. This study investigated the prevalence of Escherichia coli in imported 1-day-old chicks, ducklings, and turkey poults. Materials and Methods: The liver, heart, lungs, and yolk sacs of 148 imported batches of 1-day-old flocks (chicks, 45; ducklings, 63; and turkey poults, 40) were bacteriologically examined for the presence of E. coli. Results: We isolated 38 E. coli strains from 13.5%, 6.7%, and 5.4% of imported batches of 1-day-old chicks, ducklings, and turkey poults, respectively. They were serotyped as O91, O125, O145, O78, O44, O36, O169, O124, O15, O26, and untyped in the imported chicks; O91, O119, O145, O15, O169, and untyped in the imported ducklings; and O78, O28, O29, O168, O125, O158, and O115 in the imported turkey poults. The E. coli isolates were investigated for antibiotic resistance against 16 antibiotics using the disk diffusion method and were found resistant to cefotaxime (60.5%), nalidixic acid (44.7%), tetracycline (44.7%), and trimethoprim-sulfamethoxazole (42.1%). The distribution of extended-spectrum β-lactamase (ESBL) and ampC β-lactamase genes was blaTEM (52.6%), blaSHV (28.9%), blaCTX-M (39.5%), blaOXA-1 (13.1%), and ampC (28.9%). Conclusion: Imported 1-day-old poultry flocks may be a potential source for the dissemination of antibiotic-resistant E. coli and the ESBL genes in poultry production

Virulence Range and New Pathological Pictures of Salmonella enteridits and Salmonella typhimurium Isolated from Ducklings in Experimental Infected Chicks

Salmonellosis is a major global pathogen in the poultry industry and is a significant public health concern. Ducks are known to be carriers of Salmonella. Therefore, monitoring salmonellosis is the most important strategy for preventing the disease. An experimental design was planned to study the pathogenicity of two Salmonella strains. One hundred and fifty chicks were divided into three groups; group one was inoculated with the Salmonella enteritidis strain, group two was inoculated with the Salmonella typhimurium strain, and group three was UN inoculated. Symptoms, postmortem lesions and mortality rate were recorded. The chick growth performance parameters were also determined. Using ANOVA for statistical analysis, there was a significant difference in body weight, body gain, feed consumption, and feed conversion ratio between the two infected groups and the blank group (uninoculated group). In this study, the prevalence of Salmonella enteritidis was (1.73%) and Salmonella typhimurium (0.43%) in imported ducklings in Egypt. Both Salmonella strains were subjected to an antimicrobial sensitivity test. It showed that Salmonella enteritidis had a 60% antimicrobial resistance profile and Salmonella typhimurium had a 20% antimicrobial resistance profile. Furthermore, genotypic characterization was performed and the seven virulence genes(stn, avrA, sopB, ompF, invA, Mgtc, Ssaq) were found. New pathological lesions of Salmonella infection were discovered, such as skull hemorrhage at 3 days and 6 days of age, and a liver similar to a button shape in necropsied infected chicks with Salmonella typhimurium at 21 days of age. Furthermore, hemorrhagic spots were observed on the duodenum. In the presence of Salmonella, Clostridium perferingens was discovered in a bacteriological investigation of duodenal lesions samples from infected chicks. At 30 days of age, administration of acetic acid (1%) as an alternative tool for controlling Salmonella. In conclusion, salmonellosis is a risk factor for necrotic enteritis, and using acetic acid to eliminate salmonella infection is insufficient