Avian Colibacillosis Research Articles

Effect of Solid-State Fermentation Products of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans on Growth Performance of Broilers and Prevention of Avian Colibacillosis.

This study investigates the impact of the solid-state fermentation products of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans (LCBs) on the growth characteristics, immune function, intestinal morphology, cecum microbial community, and prevention of avian colibacillosis in broilers. One hundred and twenty Hyland Brown broilers (aged one day) were divided randomly into three groups (four replicates of ten broilers per group). (1) The CON group was fed a basal diet. (2) The MOD group was fed a basal diet. On day 40, APEC strain SX02 (1.1 × 105 CFU/g) was administered to the breasts of chickens in this group. (3) The LCBs group was fed a basal diet supplemented with fermentation products (98.5% basal diet + 0.5% Lactobacillus plantarum and Candida utilis solid-state fermentation products + 1.0% Bacillus coagulans solid-state fermentation products). On day 40, the LCBs group received the same treatment as the MOD group. The experiment lasted 43 days. This study found that the average daily gain (ADG) of the LCBs group was significantly higher than that of the MOD group (p < 0.05), indicating that LCBs can significantly increase the ADG of broilers and improve the feed conversion ratio. Furthermore, compared to the MOD group, the heart bacterial load was significantly reduced in the LCBs group (p < 0.05), and the lesions less severe in the heart, liver, and jejunum were observed (p < 0.05). Additionally, the detection of intestinal flora showed a significant increase in the abundance of beneficial bacteria in the cecum of the LCBs group, while the number of Escherichia coli and Shigella decreased significantly. In conclusion, the solid fermentation of Lactobacillus plantarum, Candida utilis, and Bacillus coagulans can improve the growth performance of broilers while also protecting against avian pathogenic Escherichia coli infection. This demonstrates the potential usefulness of these LCBs in feed production.

Study of Avian Colibacillosis and Antimicrobial Resistance in Escherichia coli from Chickens (Gallus domesticus) in Maiduguri, Borno State, Nigeria

Bioethanol is a renewable, colorless, less toxic, and readily biodegradable form of fuel from biological sources; that can be used for heat, electricity, and fuel. In this research study, an alternative feedstock known as microalgae was used. Bioethanol was produced from microalgae using enzymatic hydrolysis. The microalgae was pretreated with acid and alkaline. Hydrolysis was carried out by Aspergillus niger and Saccharomyces cerevisiae was used for fermentation. The concentration of the Bioethanol produced was determined using acid Potassium Dichromate. After hydrolysis, the reducing sugar concentration was determined using Dinitrosalicylic acid (DNS) colorimetric method. Results showed that there was significant difference at p&lt;0.05 in the yields of the reducing sugar produced between acid and alkaline pretreated samples after microbial hydrolysis. Microbial hydrolysis of alkaline pretreated samples was higher in yield. Also, the results showed that the highest bioethanol yield of 0.142ml/l was obtained after alkaline pretreated hydrolysates were fermented with Saccharomyces cerevisiae at 40°C and pH of 6.0. The highest bioethanol yield of 0.116ml/l was obtained from the acid pretreated hydrolysates at 35°C and pH of 5.5. The volatile compounds produced by alkaline pre-treated samples using Gas Chromatography Mass Spectroscopy (GCMS) shows that Ethanol and Di-n-decylsulfone were the highest volatile compounds with peak areas of 30.70% and 41.87% respectively while 2-hydroxy-1-(hydroxymethyl) ethyl ester (linolenic acid) was found to be least volatile compound with peak area of 3.34%.Tthe volatile compounds produced by acid pre-treated samples shows that 2-ethyl-2-methyltridecanol was the highest volatile compound with peak area of 23.00% while Heptacosane was found to be the least volatile compound with peak area of 1.90%. Therefore bioethanol production using alkaline pretreated samples produced a large quantity of bioethanol and may be a preferred method. This study concluded that Microalgae is a suitable substrate for bioethanol production and hydrolyzing with Aspergillus niger and optimization of fermentation conditions at pH 6.0 and 40oC yielded higher bioethanol concentration.

Exploiting membrane vesicles derived from avian pathogenic Escherichia coli as a cross-protective subunit vaccine candidate against avian colibacillosis

Avian pathogenic Escherichia coli (APEC) is a notable pathogen that frequently leads to avian colibacillosis, posing a substantial risk to both the poultry industry and public health. The commercial vaccines against avian colibacillosis are primarily inactivated vaccines, but their effectiveness is limited to specific serotypes. Recent advances have highlighted bacterial membrane vesicles (MV) as a promising candidate in vaccine research. How to produce bacterial MVs vaccines on a large scale is a significant challenge for the industrialization of MVs. The msbB gene encodes an acyltransferase and has been implicated in altering the acylation pattern of lipid A, leading to a decrease in lipid A content in lipopolysaccharides (LPS). Here, we evaluated the immunoprotective efficacy of MVs derived from the LPS low-expressed APEC strain FY26ΔmsbB, which was an APEC mutant strain with a deletion of the msbB gene. The nitrogen cavitation technique was employed to extract APEC MVs, with results indicating a significant increase in MVs yield compared to that obtained under natural culture. The immunization effectiveness was assessed, revealing that FY26ΔmsbB MVs elicited an antibody response of laying hens and facilitated bacterial clearance. Protective efficacy studies demonstrated that immunization with FY26ΔmsbB MVs conferred the immune protection in chickens challenged with the wild-type APEC strain FY26. Notably, LPS low-carried MVs recovered from the mutant FY26ΔmsbB also displayed cross-protective capabilities, and effectively safeguarding against infections caused by O1, O7, O45, O78, and O101 serotypes virulent APEC strains. These findings suggest that MVs generated from the LPS low-expressed APEC strain FY26ΔmsbB represent a novel and empirically validated subunit vaccine for the prevention and control of infections by various APEC serotypes.

High-throughput Oxford Nanopore sequencing-based approach for the multilocus sequence typing analysis of large-scale avian Escherichia coli study in Mississippi

Avian pathogenic Escherichia coli (APEC) cause avian colibacillosis and accurately distinguishing infectious isolates is critical for controlling its transmission. Multilocus sequence typing (MLST) is an accurate and efficient strain identification method for epidemiological surveillance. This research aimed to develop a fast and high-throughput workflow that simultaneously sequences the Achtman typing scheme's 7 housekeeping genes of multiple E. coli isolates using the Oxford Nanopore Technologies (ONT) platform for large-scale APEC study. E. coli strains were isolated from poultry farms, the housekeeping genes were amplified, and amplicons were sequenced on an R9.4 MinION flow cell using the Nanopore GridION sequencer (ONT, Oxford, UK) following the initial workflow (ONT-MLST). Moreover, the workflow was revised by introducing large-scale DNA extraction and multiplex PCR into the ONT-MLST workflow and applied to 242 new isolates, 18 isolates from the previous workflow, and 5 ATCC reference strains using Flongle flow cell on the Nanopore MinION Mk1C sequencer (ONT, Oxford, UK). Finally, the sequence type (ST) results of the 308 isolates collected from infected chickens and poultry farm environments were reported and analyzed. Data indicated that E. coli belonging to ST159, ST8578, and ST355 have the potential to infect multiple organs in broiler. In addition, zoonotic STs, ST69, ST10, ST38, and ST131, were detected from poultry farms. With the advantages of the high throughput of ONT, this study provides a rapid workflow for large-scale E. coli typing and identified frequently isolated sequence types related to APEC infection in poultry.

Assessment of antibacterial activity, modes of action, and synergistic effects of Origanum vulgare hydroethanolic extract with antibiotics against avian pathogenic Escherichia coli

This study evaluates the antibacterial effectiveness of Origanum vulgare hydroethanolic extract, both independently and in combination with antibiotics, against Escherichia coli strains associated with avian colibacillosis—a significant concern for the poultry industry due to the rise of antibiotic-resistant E. coli. The urgent demand for new treatments is addressed by analyzing the extract's phytochemical makeup via High-Performance Liquid Chromatography (HPLC), which identified sixteen phenolic compounds. Antibacterial activity was determined through agar diffusion and the measurement of minimum inhibitory and bactericidal concentrations (MIC and MBC), showing moderate efficacy (MIC: 3.9 to 7.8 mg/mL, MBC: 31.2 to 62.4 mg/mL). Combining the extract with antibiotics like ampicillin and tetracycline amplified antibacterial activity, indicating a synergistic effect and highlighting the importance of combinatory treatments against resistant strains. Further analysis revealed the extract's mechanisms of action include disrupting bacterial cell membrane integrity and inhibiting ATPase/H+ proton pumps, essential for bacterial survival.Moreover, the extract effectively inhibited and eradicated biofilms, crucial for preventing bacterial colonization. Regarding cytotoxicity, the extract showed no hemolytic effect at 1 to 9 mg/mL concentrations. These results suggest Origanum vulgare extract, particularly when used with antibiotics, offers a promising strategy for managing avian colibacillosis, providing both direct antibacterial benefits and moderating antibiotic resistance, thus potentially reducing the economic impact of the disease on the poultry industry.

Epidemiological study and identification of Escherichia coli strains associated with clinical events in Avian farming

Avian pathogenic Escherichia coli (APEC) represents a major challenge for the poultry industry, causing significant economic losses. This problem is exacerbated by the misuse use of antibiotics in Veterinary Medicine, leading to the emergence of resistant strains and thus creating a significant risk to Public Health. This study, carried out on 38 poultry farms in Algeria, involved the collection of 200 samples for the isolation of E. coli strains. The resistance of these strains to frequently used antibiotics was assessed using the agar diffusion method. Multiple Correspondence Analysis (MCA) was used to determine potential risk factors. The obtained results revealed that E. coli was present in 30% of samples. Alarming levels of resistance were observed against Tetracycline (81.6%), Ampicillin (78.3%), Ciprofloxacin (68.3%) and Nalidixic acid (60%). Stressful environmental conditions in poultry houses, such as temperature variations, high humidity, poor ventilation and stocking density were identified as key factors in the development of avian colibacillosis. In conclusion, the current study highlights the urgent need to strictly monitor and regulate the use of antibiotics in Veterinary Medicine and improve animal welfare in order to minimize the risk it pose to Public Health originated in the farms. In addition, it is essential that farmers maintain optimal environmental conditions in chicken rearing.

CATH-2-derived antimicrobial peptide inhibits multidrug-resistant Escherichia coli infection in chickens.

Avian colibacillosis (AC), caused by infection with Escherichia coli (E. coli), is a major threat to poultry health, food safety and public health, and results in high mortality and significant economic losses. Currently, new drugs are urgently needed to replace antibiotics due to the continuous emergence and increasing resistance of multidrug-resistant (MDR) strains of E. coli caused by the irrational use of antibiotics in agriculture and animal husbandry. In recent years, antimicrobial peptides (AMPs), which uniquely evolved to protect the host, have emerged as a leading alternative to antibiotics in clinical settings. CATH-2, a member of the antimicrobial cathelicidin peptide family, has been reported to have antibacterial activity. To enhance the antimicrobial potency and reduce the adverse effects on animals, we designed five novel AMPs, named C2-1, C2-2, C2-3, C2-4 and C2-5, based on chicken CATH-2, the secondary structures of these AMPs were consistently α-helical and had an altered net charge and hydrophobicity compared to those of the CATH-2 (1-15) sequences. Subsequently, the antimicrobial activities of CATH-2 (1-15) and five designed peptides against MDR E. coli were evaluated in vitro. Specifically, C2-2 showed excellent antimicrobial activity against either the ATCC standard strain or veterinary clinical isolates of MDR E. coli, with concentrations ranging from 2-8 μg/mL. Furthermore, C2-2 maintained its strong antibacterial efficacy under high temperature and saline conditions, demonstrating significant stability. Similarly, C2-2 retained a high level of safety with no significant hemolytic activity on chicken mature red blood cells or cytotoxicity on chicken kidney cells over the concentration range of 0-64 μg/mL. Moreover, the administration of C2-2 improved the survival rate and reduced the bacterial load in the heart, liver and spleen during MDR E. coli infection in chickens. Additionally, pathological damage to the heart, liver and intestine was prevented when MDR E. coli infected chickens were treated with C2-2. Together, our study showed that C2-2 may be a promising novel therapeutic agent for the treatment of MDR E. coli infections and AC.

Virulence-associated genes in faecal and clinical Escherichia coli isolates cultured from broiler chickens in Australia.

A healthy chicken's intestinal flora harbours a rich reservoir of Escherichia coli as part of the commensal microbiota. However, some strains, known as avian pathogenic E. coli (APEC), carry specific virulence genes (VGs) that enable them to invade and cause extraintestinal infections such as avian colibacillosis. Although several VG combinations have been identified, the pathogenic mechanisms associated with APEC are ill-defined. The current study screened a subset of 88 E. coli isolates selected from 237 pre-existing isolates obtained from commercial poultry flocks in Australia. The 88 isolates were selected based on their enterobacterial repetitive intergenic consensus (ERIC) and antimicrobial resistance (AMR) profiles and included 29 E. coli isolates cultured from chickens with colibacillosis (referred to as clinical E. coli or CEC) and 59 faecal E. coli (FEC) isolates cultured from clinically healthy chickens. The isolates were screened for the presence of 35 previously reported VGs. Of these, 34 were identified, with iucA not being detected. VGs focG, hlyA and sfa/foc were only detected in FEC isolates. Eight VGs had a prevalence of 90% or above in the CEC isolates. Specifically, astA (100%); feoB (96.6%); iutA, iss, ompT, iroN and hlyF (all 93.1%); and vat (89.7%). The prevalence of these were significantly lower in FEC isolates (astA 79.7%, feoB 77.9%, iutA 52.5%, iss 45.8%, ompT 50.9%, iroN 37.3%, hlyF 50.9% and vat 42.4%). The odds ratios that each of these eight VGs were more likely to be associated with CEC than FEC ranged from 7.8 to 21.9. These eight VGs may be used to better define APEC and diagnostically detect APEC in Australia. Further investigations are needed to identify the roles of these VGs in pathogenicity.

Study of the Prevalence of E. Coli in Poultry and Selection of Specific Bacteriophages

Avian colibacillosis is one of the leading causes of mortality and morbidity associated with economic losses in the industry throughout the world. The problem is enhanced by the growing resistance of pathogenic pathogens towards antibiotics. This situation has strongly stimulated a renewal of scientists’ interest in bacteriophages (phages), viruses of bacteria. They are abundant in nature, and accompany bacteria in each environment they colonize, including human microbiota. The aim of our work was to study the prevalence of pathogenic E-coli in poultry, isolate, characterize, and evaluate the potential use of isolated bacteriophages to control E. coli infections in poultry. 35 pathogenic E. coli strains were isolated from 90 sample of poultry excreta. The most common phylogenetic groups were A1 (25%), A2 (15%), B1 (35%), and B2 (25%). E. coli was 100% resistant to five antibiotics (Bacitracin, Clindamycin, Carbapenem, Cephalexin, Clarithromycin). E. coli was least resistant to oxacillin (40%), followed by tetracycline (48.6%). The prevalence of multidrug resistance was 91.4%. Such high levels of resistance in E. coli isolated from poultry excreta could pose a serious threat to humans. Five novel phages against pathogenic E. coli were isolated from sewage water and characterized in vitro. The electron microscopic analysis showed that two phages belonged to the Myoviridae family and three bacteriophages belonged to the Siphoviridae family, in the order Caudovirales

Genome-wide transcriptional profiling and functional analysis of long noncoding RNAs and mRNAs in chicken macrophages associated with the infection of avian pathogenic E. coli

BackgroundAvian pathogenic E. coli (APEC) can cause localized or systemic infections, collectively known as avian colibacillosis, resulting in huge economic losses to poultry industry globally per year. In addition, increasing evidence indicates that long non-coding RNAs (lncRNAs) play a critical role in regulating host inflammation in response to bacterial infection. However, the role of lncRNAs in the host response to APEC infection remains unclear.ResultsHere, we found 816 differentially expressed (DE) lncRNAs and 1,798 DE mRNAs in APEC infected chicken macrophages by RNAseq. The identified DE lncRNA-mRNAs were involved in Toll like receptor signaling pathway, VEGF signaling pathway, fatty acid metabolism, phosphatidylinositol signaling system, and other types of O-glycan biosynthesis. Furthermore, we found the novel lncRNA TCONS_00007391 as an important immune regulator in APEC infection was able to regulate the inflammatory response by directly targeting CD86.ConclusionThese findings provided a better understanding of host response to APEC infection and also offered the potential drug targets for therapy development against APEC infection.

Effect of caffeic acid grafted chitosan loaded quercetin lyophilized powder formulation on avian colibacillosis and tissue distribution.

Quercetin (QR), recognized as a natural antibacterial ingredient, has found widespread application in the poultry industry. This study investigated the bacteriostatic mechanism and evaluated the in vivo inhibitory impact of caffeic acid-grafted chitosan self-assembled micelles loaded quercetin (CA-g-CS/QR) on avian Escherichia coli (E. coli). The findings indicate that the bactericidal mechanism of CA-g-CS/QR exhibits enhanced efficacy compared to QR alone, disrupting bacterial cell walls, disassembling biofilm structures, and impeding essential components necessary for bacterial growth. Following an avian E. coli attack in broilers, CA-g-CS/QR demonstrated the capacity to enhance the population of beneficial bacteria while concurrently decreasing harmful bacteria within the intestinal tract. Moreover, within 3 days of oral administration of CA-g-CS/QR, a significant decrease in Escherichia spp. count was evident, resulting in the restoration of broilers to a healthy state. CA-g-CS/QR proved to be a significant and more efficacious solution than QR alone for avian E. coli disease. Furthermore, CA-g-CS/QR displayed a broader distribution range and higher concentration within the body. Ten metabolites have been identified in the liver for both QR and CA-g-CS/QR. In conclusion, CA-g-CS/QR has demonstrated a notable capacity to enhance in vitro and in vivo bacterial inhibitory effects, providing foundation for the clinical application of QR in combating avian E. coli infections in broilers.

Characteristics, Whole-Genome Sequencing and Pathogenicity Analysis of Escherichia coli from a White Feather Broiler Farm.

Avian colibacillosis, caused by avian Escherichia coli (E. coli), has historically been one of the most prevalent infectious diseases in large-scale poultry production, causing growth delays and mortality in chickens, resulting in huge economic losses. In recent years, the widespread use of antibiotics has led to the emergence of multidrug resistance in E. coli as a significant global problem and long-term challenge. Resistant E. coli can be transmitted to humans through animal products or the environment, which presents significant public health concerns and food safety issues. In this study, we analyzed the features of 135 E. coli strains obtained from a white feather broiler farm in Shandong, China, including antimicrobial susceptibility tests, detection of class 1 integrons, drug resistance genes, virulence genes, and phylogenetic subgroups. It is particularly worrying that all 135 E. coli strains were resistant to at least five antibiotic agents, and 100% of them were multidrug-resistant (MDR). Notably, the resistance genes of blaTEM, blaCTX-M, qnrS, aaC4, tetA, and tetB exhibited a high prevalence of carriage among the tested resistance genes. However, mcr-2~mcr-9 were not detected, while the prevalence of mcr-1 was found to be 2.96%. The most common virulence genes detected were EAST1 (14.07%, encoding enterotoxins) and fyuA (14.81%, encoding biofilm formation). Phylogenetic subgroup analysis revealed that E. coli belonging to groups B2 and D, which are commonly associated with high virulence, constituted 2.22% and 11.11%, respectively. The positive rate of class 1 integrons was 31.1%. Whole-genome sequencing (WGS) and animal experiments were performed on a unique isolated strain called 21EC78 with an extremely strong membrane-forming capacity. The WGS results showed that 21EC78 carried 11 drug resistance genes and 16 virulence genes. Animal experiments showed that intraperitoneal injection with 2 × 105 CFU could cause the death of one-day-old SPF chickens in 3 days. However, the mortality of Luhua chickens was comparatively lower than that of SPF chickens. This study reports the isolation of multidrug-resistant E. coli strains in poultry, which may pose a potential threat to human health via the food chain. Furthermore, the findings of this study enhance our comprehension of the frequency and characteristics of multidrug-resistant E. coli in poultry farms, emphasizing the urgent need for improved and effective continuous surveillance to control its dissemination.

CTX-M-55-type ESBL-producing fluoroquinolone-resistant Escherichia coli sequence type 23 repeatedly caused avian colibacillosis in Kagoshima Prefecture, Japan

The production of expanded-spectrum beta-lactamases (ESBLs) and fluoroquinolone resistance in Enterobacteriaceae has become a global concern. The aim of this study was to investigate the spread of ESBL-producing and fluoroquinolone-resistant avian pathogenic Escherichia coli (APEC) in Kagoshima, a prefecture with the largest amount of poultry in Japan. The antimicrobial susceptibility and genetic characteristics of 228 APEC strains isolated from 57 farms in Kagoshima Prefecture, Japan, between 2005 and 2017 were analysed. Information about the companies with hatcheries connected to the farms was also collected, and the epidemiologic relatedness of APEC strains and the processes of adopting chicks were compared. Seven CTX-M-type ESBL genes, blaCTX-M-1, blaCTX-M-2, blaCTX-M-14, blaCTX-M-15, blaCTX-M-25, blaCTX-M-55, and blaCTX-M-65, were found in 60 (26.3%) of the 228 APEC strains. The ciprofloxacin-resistant strains belonged to 10 different sequence types (ST10, ST23, ST93, ST155, ST156, ST350, ST359, ST602, ST648, and ST9479), and the two ST602 strains showed remarkably high ciprofloxacin resistance (MIC: 128 µg/mL) and had amino acid mutations in GyrA (S83L and D87N), ParC (S80I), and ParE (E460A). A CTX-M-55-type ESBL-producing fluoroquinolone-resistant Og78-ST23 strain was isolated multiple times over two years on a farm. Furthermore, epidemiologically closely related strains were isolated from different farms that used the same common hatcheries. APEC is often transferred from hatcheries to farms via healthy chicks, and the prudent use of antimicrobials and careful monitoring of resistant strains on poultry farms and hatcheries are important in preventing the selection and spread of high-risk APEC strains such as CTX-M-55-type ESBL-producing Og78-ST23.

A Comparative Study on Two Infection Models of Colibacillosis in Broilers: Clinical Features, Pathogenesis, and Response to Therapy.

Knowledge on the characteristics of different infection models of avian colibacillosis is mandatory for performing reliable experimental studies. This study compares the characteristics of two infection models of colibacillosis (by intratracheal and subcutaneous routes) in broilers. Broilers (125), 35 days old, were randomly allocated to four experimental (n = 20) and three control (n = 15) groups: 1) negative control; 2) intratracheal infection control; 3) subcutaneous infection control; 4) intratracheal infection (IT); 5) intratracheal infection with florfenicol administration (ITF); 6) subcutaneous infection (SC); and 7) subcutaneous infection with florfenicol administration (SCF). Clinical signs and mortalities were recorded, and gross examination of carcasses was performed. On days 3 and 5 postchallenge (PC), 5 birds/group were euthanatized. Despite similar onset and type of clinical signs and mortality of birds in the IT and SC groups, especially on early days PC, birds in the SC group had lower body weight than IT birds. Lung injuries were more prominent in the IT group than the SC group. Birds of the SC group showed cellulitis and severely injured kidneys in contrast to the IT group. On day 3 PC, the number of positive heart blood samples for bacterial growth in the ITF group was half that of the IT group, whereas SC and SCF birds had similar positive numbers. Infection by the subcutaneous route results in more severe disease. The intratracheal route produces results more similar to colibacillosis under field conditions with regard to clinical signs, pathogenesis, and response to therapy, albeit it is more difficult to perform than a subcutaneous challenge.

A comparative study on histopathological features of duodenum, jejunum, and ileum from broiler chicken with avian pathogenic Escherichia coli infection

Avian colibacillosis is a high-prevalence chicken farm disease caused by avian pathogen-ic Escherichia coli (APEC). It is necessary to identify and look up to the bacterial activity to stave off a decline in chicken performance. This study aimed to identify and determine the pathogenetic activity of APEC within broiler chickens on the duodenum, jejunum, and ileum histopathology comparison. The samples were collected from a total of 40 broilers, obtained from a highest-colibacillosis historical chicken house, identified with bacterial isolation swab-technique, and analyzed with nonparametric statistical lesions scoring. The identification result showed 82.5% E. coli with a 62.5% APEC con-firmed. The histopathological feature showed a significant difference (p&lt;0.05), where the jejunum ap-peared to encounter the most significant damage compared to the duodenum and ileum. Thus, it is con-cluded that APEC field infection varies in characteristics and pathogenicity, which affects intestinal organs based on the histology changes, mainly in the jejunum part of the intestine.

Prevalence and Risk Factors of Multidrug Resistant Salmonella spp and Escherichia coli in Poultry from Doba Commune, Chad

Introduction: Avian colibacillosis and salmonellosis are diseases that have a major economic impact on poultry farming and public health worldwide. This study aims to determine the prevalence and associated risk factors of multidrug resistant Salmonellosis and Escherichia coli in Doba commune, Chad&#x0D; Methodology: This is an experimental study that took place in Doba, the survey of which was carried out among 41 poultry farming households in 15 districts. The samples of faeces were taken with an average of 13 samples per district and were cultured at the IRED Laboratory in N'Djamena. The strains of Salmonella spp and E. coli isolated were identified by the Enterosystem 18R gallery. Their antibiotic sensitivities were tested against 12 selected antibiotics. The Epi Info 7 ™ software was used to perform the statistical analyses.&#x0D; Results: For this study, 193 samples of faeces were cultured. However, 24 (12.44%) strains were isolated and identified, including 13 (6.74%) Salmonella spp and 11 (5.70%) Escherichia coli. These strains of Salmonella spp were more sensitive to Imipenem (92.31%). They were more resistant to Clotrimazole (84.62%). For E. coli, the highest sensitivity was observed in Imipenem (76.92%). It was more resistant to Amoxicillin + Clavunalic Acid (AMC) 69.23%. Non-compliance with food hygiene, lack of maintenance of the habitat, non-compliance with prophylactic and sanitary measures were the risk factors most linked to the presence of salmonellosis and E. coli. According to the profile, the resistance of Salmonella spp to antibiotics was more observed in Béraba (23.08%) and that of Escherichia coli in Bédokassa (27.27%).&#x0D; Conclusion: This study allowed the identification of multidrug resistant Salmonella spp and Escherichia coli in poultry farming households. Non-compliance with food hygiene, lack of maintenance of the habitat, non-compliance with prophylactic and sanitary measures were the risk factors most linked to the presence of salmonellosis and E. coli.

Therapeutics and prophylactic efficacy of novel lytic Escherichia phage vB_EcoS_PJ16 against multidrug-resistant avian pathogenic E. coli using in vivo study.

Avian pathogenic Escherichia coli (APEC) is the causative agent of avian colibacillosis, which causes significant economic losses to the poultry industry. The growing resistance of bacteria to antibiotics is a major global public health concern. However, there is limited data on the efficacy of phage therapy in effectively controlling and treating APEC infections. In this study, a novel lytic Escherichia phage, vB_EcoS_PJ16, was isolated from poultry farm wastewater and characterized in both in vitro and in vivo conditions. Transmission electron microscopy analysis revealed the presence of an icosahedral head and a long non-contractile tail, classifying the phage under the Caudoviricetes class. Host range determination showed that Escherichia phage vB_EcoS_PJ16 exhibited lytic activity against multiple strains of pathogenic E. coli, while no significant signs of lysis for Klebsiella pneumoniae, Salmonella Typhimurium, Listeria monocytogenes, and Staphylococcus aureus. Biophysical characterization revealed that the isolated phage was sturdy, as it remained viable for up to 300days at temperatures of 30°C, 37°C, and 42°C and for up to 24h at pH 5 to 11, with only minor changes in titer. Kinetic analysis at multiplicity of infection (MOI) 0.1 showed a latency period of about 20min and a burst size of 26.5 phage particles per infected cell for phage vB_EcoS_PJ16. Whole genome sequencing unveiled that the phage vB_EcoS_PJ16 genome consists of a double-stranded linear DNA molecule with 57,756bp and a GC content of 43.58%. The Escherichia phage vB_EcoS_PJ16 genome consisted of 98 predicted putative ORFs, with no transfer RNA identified in the genome. Among these 98 genes, 34 genes were predicted to have known functions. A significant reduction in APEC viability was observed at MOI 100 during in vitro bacterial challenge tests conducted at different MOIs (0.01, 1, and 100). In vivo oral evaluation of the isolated phage to limit E. coli infections in day-old chicks indicated a decrease in mortality within both the therapeutic (20%) and prophylactic (30%) groups, when compared to the control group. The findings of this study contribute to our current knowledge of Escherichia phages and suggest a potentially effective role of phages in the therapeutic and prophylactic control of antibiotic-resistant APEC strains.

In ovo administration of a phage cocktail partially prevents colibacillosis in chicks

Avian pathogenic Escherichia coli (APEC) causes colibacillosis, the main bacterial disease in poultry leading to significant economic losses worldwide. Antibiotic treatments favor the emergence of multidrug-resistant bacteria, and preventive measures are insufficient to control the disease. There is increasing interest in using the potential of bacteriophages, not only for phage therapy but also for prevention and biocontrol. This study aimed to evaluate the efficacy of a phage cocktail administered in ovo to prevent avian colibacillosis in chicks. When 4 different phages (REC, ESCO3, ESCO47, and ESCO58), stable under avian physiological conditions, were combined and inoculated at 17 embryogenic days (ED), they were transmitted to the newly hatched chicks. In a second trial, the 4-phage cocktail was inoculated into the allantoic fluid at ED16 and after hatch 1-day-old chicks were challenged with the O2 APEC strain BEN4358 inoculated subcutaneously. Two phages (REC and ESCO3) were still detected in the ceca of surviving chicks at the end of the experiment (7-days postinfection). Chicks that received the phages in ovo did not develop colibacillosis lesions and showed a significant decrease in intestinal BEN4358 load (8.00×107 CFU/g) compared to the challenged chicks (4.52×108 CFU/g). The majority of the reisolated bacteria from the ceca of surviving chicks had developed full resistance to ESCO3 phage, and only 3 were resistant to REC phage. The partially or complete resistance of REC phage induced a considerable cost to bacterial virulence. Here, we showed that phages inoculated in ovo can partially prevent colibacillosis in 1-wk-old chicks. The reduction in the APEC load in the gut and the decreased virulence of some resistant isolates could also contribute to control the disease.

Detection of high-risk Avian Pathogenic Escherichia coli (APEC) isolated from broilers in São Paulo, Brazil.

Some high-risk Avian Pathogenic Escherichia coli (APEC) clones have been associated with increased economic losses caused by avian colibacillosis. They may represent an additional food consumption concern due to the potential zoonotic role causing urinary tract infections mainly related to E. coli ST73 and ST95 lineages. This study aimed to characterize APEC isolated from slaughterhouse carcasses presenting lesions compatible with avian colibacillosis. We analyzed about 6500 broilers carcasses, and 48 showed lesions consistent with colibacillosis. Forty-four strains of E. coli were isolated, with 77.27% (n = 34/44) classified as APEC. The isolates belonged to the phylogenetic groups B2 (41.17%, n = 14/34), G (20.59%, n = 7/34), A (17.65%, n = 6/34), B1 (8.82%, n = 3/34), and E (5.88%, n = 2/34). Determining the phylogenetic group of 5.88% (n = 2/34) of the strains was impossible. Moreover, 20.59% (n = 7/34) were positive to the clonal groups ST117, 8.82% (n = 3/34) to ST95, and 8.82% (n = 3/34) were classified as belonging to serogroup O78 by PCR screening. Strains of APEC from O78 serogroup and ST117 are considered high-risk clones for poultry, and our data reinforced the need for surveillance of these pathogens in poultry farms and slaughterhouses.

The manganese carbonyl complex [Mn(CO)3(tqa-κ3N)]Br: A novel antimicrobial agent with the potential to treat avian pathogenic Escherichia coli (APEC) infections

The development of alternatives to antibiotics is essential for the treatment of animal infections and as a measure to reduce the selective pressure on antibiotics that are critical for human medicine. Metal complexes have been highlighted for their antimicrobial activity against several bacterial pathogens. In particular, manganese carbonyl complexes have shown efficacy against multidrug-resistant Gram-negative pathogens, and relatively low cytotoxicity against avian macrophages and in wax moth larval models. They are thus potential candidates for deployment against Avian Pathogenic Escherichia coli (APEC), the aetiological agent of avian colibacillosis, which results in severe animal welfare issues and financial losses worldwide. This study aimed to determine the efficacy of [Mn(CO)3(tqa-κ3N)]Br in Galleria mellonella and chick models of infection against APEC. The results demonstrated in vitro and in vivo antibacterial activity against all antibiotic-resistant APEC test isolates screened in the study.