Antimicrobial resistance pattern of avian pathogenic Escherichia coli with detection of extended-spectrum ß-lactamase-producing isolates in broilers in east Algeria.

Background and Aim: Avian pathogenic Escherichia coli (APEC) is the causative agent of colibacillosis, one of the most prevalent bacterial diseases responsible for significant economic losses in the poultry industry worldwide. This study aimed to assess the antimicrobial resistance (AMR) patterns of APEC isolates recovered from poultry in east Algeria and estimate the prevalence of extended-spectrum ß-lactamase (ESBL)-producing isolates. Materials and Methods: In the slaughterhouse of Batna City (Algeria), livers indicating colibacillosis were sampled from 204 suspected carcasses with growth retardation and generalized congestion. Escherichia coli isolation and identification were performed on MacConkey agar using conventional methods and the API 20E system. Antimicrobial resistance susceptibility was tested by the disk diffusion method according to the Clinical Laboratory Standards Institute Guidelines. Extended-spectrum ß-lactamase detection was carried out using the double-disk confirmation test. Results: One hundred sixty E. coli isolates were recovered (one isolate per sample). Avian pathogenic Escherichia coli isolates showed high levels of resistance to ampicillin and tetracycline (100%), nalidixic acid (95%), ofloxacin (93.75%), doxycycline (91.87), ciprofloxacin (87.50%), trimethoprim/sulfamethoxazole (62.50%), gentamycin (32.50%), chloramphenicol (27.50%), amoxicillin/clavulanic acid (16.25%), colistin (14.37%), and nitrofurantoin (10.62%). All strains were multidrug-resistant to at least three antibiotics, and more than half (52.52%) of the isolates were resistant to at least seven antibiotics. All isolates were susceptible to ceftriaxone, ceftazidime, and aztreonam. Two E. coli isolates were ESBL producers (1.25%). Conclusion: Avian pathogenic Escherichia coli resistance to most antimicrobial agents used in poultry may lead to antimicrobial therapy failure.

Indicators and risk factors of infectious laryngotracheitis in layer hen flocks in Algeria.

BACKGROUND AND AIM: Since 2017, there have been epidemics with respiratory disorders in the laying hen farms in Algeria, as signs and lesions, respiratory difficulties, and hemorrhagic tracheitis, which closely like laryngotracheitis. This study aimed to analyze the epidemiological, serological, and clinical indicators, as well as the risk factors, of infectious laryngotracheitis (ILT) in layer hen flocks in Algeria. MATERIALS AND METHODS: A total of 1728 layer hens were sampled randomly from 48 poultry houses. Blood samples were collected from each hen at the wing vein area, and an indirect enzyme-linked immunosorbent assay was done using an IDvet® kit. RESULTS: The flocks showed 56.25% seroprevalence. Clinical signs and gross lesions of ILT suspect cases included respiratory signs characterized by hemorrhagic tracheitis and sinusitis; conjunctivitis; egg drop; and a low mortality rate varying from 5% to 20%. Statistical analyses showed the effect of risk factors on the seropositivity for ILT in 48 layer flocks. When the vaccination was not applied, flocks were significantly more seropositive by 54% (odds ratio OR=1.54, p=0.01) compared to vaccinated flocks. Furthermore, flocks with poor hygiene were more seropositive by 68% (OR=1.68, p=0.002) compared to those with good hygiene. Finally, flocks with decreased egg production between 10% and 30% were significantly more seropositive by 42% (OR=1.42, p=0.04) than those with egg production >30%. CONCLUSION: The serological survey revealed anti-ILT virus antibodies, signifying the circulation of this virus in layer hen farms in Algeria. Correct vaccination protocol, strict biosecurity measures, rapid diagnosis, and detection of latent carriers are necessary to control and eradicate the disease from layer farms.

Detection of methicillin-resistant coagulase-negative staphylococci and PVL/mecA genes in cefoxitin-susceptible Staphylococcus aureus (t044/ST80) from unpasteurized milk sold in stores in Djelfa, Algeria.

This study was designed to determine antimicrobial resistance phenotypes and genotypes and virulence factors in Staphylococcus aureus and coagulase-negative staphylococci (CNS) in unpasteurized milk sold in Djelfa, Algeria. Eighty-two unpasteurized cow milk samples were randomly obtained from 82 retail stores in Djelfa and tested to detect staphylococci. Species were identified by biochemical tests and MALDI-TOF. Antimicrobial resistance phenotypes and genotypes were determined by disk diffusion test, PCR, and sequencing. The Staph. aureus isolates were subjected to spa typing, multilocus sequence typing, and detection of virulence genes and the scn gene by PCR and sequencing. Forty-five (54.9%) milk samples were contaminated by staphylococci and 45 isolates were recovered: 10 Staph. aureus (12.2% of total samples) and 35 CNS (42.7%). Resistance to penicillin (blaZ), tetracycline (tetL/tetK), and erythromycin (ermB/msrA/ermC) were the most common phenotypes (genotypes). Three CNS were methicillin-resistant and all were mecA-positive. The Staph. aureus isolates were ascribed to the following lineages [spa type/sequence type/associated clonal complex (number of isolates)]: t267/ST479/CC479 (n = 6), t1510/ST5651/CC45 (n = 1), t359/ST97/CC97/ (n = 1), t346/ST15/CC15 (n = 1), and t044/ST80 (n = 1). The mecA gene was detected in the cefoxitin-susceptible t044/ST80 isolate and co-harbored the lukF/lukS-PV and scn genes. The detection of mecA-PVL-positive Staph. aureus, methicillin-resistant CNS, and multidrug-resistant staphylococcal species indicates a potentially serious health issue and reveals that unpasteurized milk sold in Djelfa city could be a potential vehicle for pathogenic and antimicrobial-resistant staphylococci.

Extended Spectrum ß-Lactamase-Producing Escherichia coli and Klebsiella pneumoniae from Broiler Liver in the Center of Algeria, with Detection of CTX-M-55 and B2/ST131-CTX-M-15 in Escherichia coli.

This study aimed to determine the prevalence and diversity of extended-spectrum ß-lactamase (ESBL)-producing and multidrug-resistant (MDR) Escherichia coli and Klebsiella pneumoniae isolates from 136 broiler livers randomly purchased in 136 retail markets in Djelfa (Algeria). Isolation was performed on Hektoen agar and bacterial identification was carried out by API20E system and Maldi-TOF-MS (matrix-assisted laser desorption/ionization time-of-flight mass spectrometry). Antimicrobial susceptibility was tested by the disk diffusion and agar dilution methods. Detection of ESBLs and other resistance and integron genes, phylogenetic grouping, and molecular typing was performed by PCR and sequencing. Seventy-eight isolates (one per positive sample) were recovered: 73 E. coli and 5 K. pneumoniae. Among E. coli, 86.3% of isolates were MDR. ESBL activity was revealed in eight E. coli and five K. pneumoniae isolates (rates of 5.9% and 3.7% in analyzed samples, respectively). ESBL genes detected among E. coli were as follows (number of isolates): blaCTX-M-15 (3), blaCTX-M-1 (3), blaCTX-M-55 (1), and blaSHV-12 (1); all ESBL-producing K. pneumoniae isolates carried the blaCTX-M-15 gene. ESBL-producing E. coli isolates were assigned to lineages (phylogroup/sequence type and number of isolates in parenthesis): A/ST48 (1), B1/ST6448 (1), B1/ST5087 (3), B1/ST23 (1), and B2/ST131 (two blaCTX-M-15 E. coli isolates). K. pneumoniae isolates were ascribed to sequence types ST2010 and ST3483. Regarding the 65 non-ESBL E. coli isolates, the most observed resistance genes were as follows: tet(A) (75%), blaTEM (57.1%), and sul2 (43.5%). Class1 integrons were revealed in seven non-ESBL E. coli isolates (10.7%) and two gene-cassette arrays were identified: dfrA1 and aadA1+dfrA1. Our study provides evidence that broiler-derived food from Center of Algeria constitutes a source of ESBL and/or MDR-producing Enterobacteriaceae, with detection of relevant ESBL genes and epidemic clones.

Serological, clinical, and risk factors of the Newcastle disease on broilers flocks in Algeria.

AIM: The work aimed at studying the serological and clinical factors, as well as the risk factors of the Newcastle disease (ND) on broilers herds in Algeria. MATERIALS AND METHODS: A sample of 1248 birds was randomly selected from 52 broiler flocks. We took blood samples from each bird at the level of the wing vein area where an indirect enzyme-linked immunosorbent assay technique was carried out through the use of an IDvet kit. RESULTS: The flocks showed 82.69% of seroprevalence. Clinically speaking, the most common symptoms were sneezing, rale, greenish diarrhea, torticollis, and motor discords. Most commonly observed postmortem lesions were the proventriculitis, tracheitis, and enteritis. Especially, the caeca are hemorrhagic. The scores show the effect of risk factors. There was a significant effect on the mortality, the hygiene and vaccination groups on antibody titers in time 2. The antibody titers were elevated in the herd that recorded a high mortality (more than 10%) compared with those which recorded a low mortality (<10%) (p=0.002). Therefore, the antibody titers were elevated in herds with bad hygiene, compared with the ones with good hygiene (p=0.04). At last, when broiler chicken were not boosted by ND vaccine, flocks appeared to be more seropositive (p=0.02). CONCLUSION: The serological survey conducted in this study provided an important scope for ND as a dominant viral disease in broilers. Many factors are responsible for the onset of these diseases; correct biosecurity measures are needed to reduce the impact of this pathology in poultry farms.