Extended-spectrum beta-lactamase (ESBL)-producing Enterobacteriaceae are a universal public health alarm frequently identified among humans, animals, and poultry. Livestock and poultry production are a possible source of multidrug-resistant microorganisms, including ESBL-producing Enterobacteriaceae, which confer antimicrobial resistance to different β-lactam antimicrobial agents. From January to May 2020, a cross-sectional study was carried out in three dairy cattle farms and four poultry farms in different districts of northern Egypt to assess the prevalence of ESBLs, AmpC beta-lactamase-producing E. coli and Klebsiella in livestock, poultry, and human contacts, and to investigate the genetic relatedness of the recovered isolates. In total, 140 samples were collected, including human fecal samples (n = 20) of workers with intimate livestock contact, cattle rectal swabs (n = 34), milk (n = 14), milking machine swabs (n = 8), rations (n = 2), and water (n = 2) from different cattle farms, as well as cloacal swabs (n = 45), rations (n = 5), water (n = 5) and litter (n = 5) from poultry farms. The specimens were investigated for ESBL-producing E. coli and Klebsiella using HiCrome ESBL media agar. The agar disk diffusion method characterized the isolated strains for their phenotypic antimicrobial susceptibility. The prevalence of ESBL-producing Enterobacteriaceae was 30.0%, 20.0%, and 25.0% in humans, cattle, and poultry, respectively. Further genotypic characterization was performed using conventional and multiplex PCR assays for the molecular identification of ESBL and AmpC genes. The majority of the ESBL-producing Enterobacteriaceae showed a multi-drug resistant phenotype. Additionally, bla_SHV was the predominant ESBL genotype (n = 31; 93.94%), and was mainly identified in humans (n = 6), cattle (n = 11), and poultry (14); its existence in various reservoirs is a concern, and highlights the necessity of the development of definite control strategies to limit the abuse of antimicrobial agents.

Enteropathogenic Escherichia coli (EPEC) is a leading cause of diarrhoeagenic diseases in humans and cattle worldwide. The emergence of multidrug-resistant (MDR) EPEC from cattle sources is a public health concern. A total of 240 samples (75 diarrhoeic calves, 150 milk samples, and 15 workers) were examined for prevalence of EPEC in three dairy farms in Egypt. Antimicrobial resistance (AMR) traits were determined by antibiogram and polymerase chain reaction (PCR) detection of β-lactamase-encoding genes, plasmid-mediated quinolone resistance genes, and carbapenemase-encoding genes. The genetic relatedness of the isolates was assessed using repetitive extragenic palindromic sequence-based PCR (REP-PCR). EPEC isolates were detected in 22.7% (17/75) of diarrhoeic calves, 5.3% (8/150) of milk samples, and 20% (3/15) of worker samples. The detected serovars were O26 (5%), O111 (3.3%), O124 (1.6%), O126 (0.8%), and O55 (0.8%). AMR-EPEC (harbouring any AMR gene) was detected in 9.2% of samples. Among isolates, bla_TEM was the most detected gene (39.3%), followed by bla_SHV (32.1%) and bla_CTX-M-1 (25%). The qnrA, qnrB, and qnrS genes were detected in 21.4%, 10.7%, and 7.1% of isolates, respectively. The bla_VIM gene was detected in 14.3% of isolates. All EPEC (100%) isolates were MDR. High resistance rates were reported for ampicillin (100%), tetracycline (89.3%), cefazolin (71%), and ciprofloxacin (64.3%). Three O26 isolates and two O111 isolates showed the highest multiple-antibiotic resistance (MAR) indices (0.85–0.92); these isolates harboured bla_SHV-12 and bla_CTX-M-15 genes, respectively. REP-PCR genotyping showed high genetic diversity of EPEC, although isolates belonging to the same serotype or farm were clustered together. Two worker isolates (O111 and O26) showed high genetic similarity (80–95%) with diarrhoeic calf isolates of matched serotypes/farms. This may highlight potential inter-species transmission within the farm. This study highlights the potential high risk of cattle (especially diarrhoeic calves) as disseminators of MDR-EPEC and/or their AMR genes in the study area. Prohibition of non-prescribed use of antibiotics in dairy farms in Egypt is strongly warranted.

Helicobacter pylori (H. pylori) and Helicobacter pullorum (H. pullorum) are frequently reported pathogens in humans and poultry, respectively. Nevertheless, the source of H. pylori is still unclear. This study aimed to detect Helicobacter spp. in chicken carcasses and to assess the antibiogram and the virulence genes of Helicobacter isolates. Three hundred chicken meat samples (100 each of chicken breast, liver, and gizzard), besides 60 swab samples from chicken processing surfaces, were collected from retail shops in Qalyubia Governorate, Egypt, and examined for the prevalence of H. pylori and H. pullorum. The 16S rRNA of three H. pylori and two H. pullorum isolates were sequenced to determine the genetic relationship between these two Helicobacter spp. Of the 300 chicken samples tested, 16 (5.33%) and 14 (4.67%) were positive for H. pylori and H. pullorum, respectively. Multiplex PCR revealed that the virulence genes vacuolating cytotoxin A (vacA)s1, cytotoxin-associated gene A (cagA), and restriction endonuclease-replacing gene A (hrgA) were detected in 66.7%, 77.8%, and 100% of H. pylori strains tested, respectively. H. pylori showed the highest resistance for clarithromycin, while H. pullorum exhibited the highest resistance towards erythromycin and ciprofloxacin. The study concluded that the chicken meat and giblets are potential sources of the virulent and antimicrobial-resistant strains of H. pylori of human origin.

Toxoplasmosis has been associated with economic and public health concerns due to abortion in humans and animals. Detection for the presence of anti-Toxoplasma gondii antibodies in sera of 174 healthy sheep (IgG) and sera of 89 aborted women (IgG, IgM, and IgG avidity) using enzyme-linked immunosorbent assay and T. gondii in 59 sheep milk samples identified by PCR. The primary risk factors for sheep and women seropositive samples were also analyzed. Relatively higher seroprevalence of T. gondii, 62.6%, was recorded in sheep than 57.3% in aborted women. The IgM/IgG antibody responses in aborted women were 42.7%, 41.6%, 15.7%, and 0% for IgM -ve/IgG -ve (negative toxoplasmosis), IgM -ve/IgG +ve (chronic toxoplasmosis), IgM +ve/IgG +ve (maternal infection) and IgM +ve/IgG -ve (acute infection), respectively. IgM and IgG-positive samples showed a low avidity rate 14.3%, denoting acute maternal infection, while a much higher rate 78.6%, denoting chronic maternal infection. Molecular investigation of T. gondii in sheep milk samples revealed that 28.8% were positive by PCR at 470-bp. The respectable significant association between many risk factors and the seroprevalence of T. gondii in humans and sheep was recorded. The current study indicates elevated toxoplasmosis antibodies in sheep and aborted women from El-Beheira and Alexandria governorates, Egypt. Additionally, it demonstrates a correlation between age, mutton intake, miscarriage, cat exposure, drinking water source, and breeding practices with seropositivity to T. gondii. These results largely confirm the need for greater in-depth toxoplasmosis epidemiological research and public health education initiatives.

Abortion results from a Toxoplasma gondii infection in both humans and mammals, causing substantial losses in terms of quality-adjusted life years and financial consequences. The incidence of toxoplasmosis in the current study in aborted women and sheep was determined using serological, immunological, and molecular assays. Blood samples were taken from sheep that appeared to be healthy and from women who had aborted in the governorates of Alexandria and El-Beheira on the west coast of Egypt. The specific immunogenetic bands in the crude tachyzoite antigens from both RH and local T. gondii strains were characterized using SDS-PAGE and immunoblotting assays. An indirect ELISA was performed to detect the prevalence in aborted women and sheep. The confirmatory molecular identification of toxoplasmosis was adopted using conventional polymerase chain reaction (cPCR). The results revealed that the two RH and local T. gondii antigens showed extensive electrophoretic similarity and identified two common immunogenic bands (67 and 58 kDa) in naturally infected human and sheep sera with T. gondii. The rate of toxoplasmosis sero-prevalence recorded by ELISA in aborted women was 60.8% (76/125), while in sheep it was 67.3% (66/98). Verified PCR results of toxoplasmosis in aborted women and sheep revealed the expected positive bands, 194 bp and 161 bp using B1 and Tox-9 & Tox-11 gene primers, respectively. This study concluded that the higher T. gondii prevalence might be responsible for abortion and economic losses in humans and sheep in our studied region. These findings reinforce the necessity of more epidemiological studies on toxoplasmosis and public health education initiatives.

Klebsiella oxytoca is an emerging opportunistic pathogen capable of causing bovine mastitis and neonatal colonization. This study provides a comprehensive molecular and phenotypic characterization of K. oxytoca across dairy supply nodes and pediatric host matrices while evaluating a probiotic-based bio-intervention strategy. K. oxytoca was isolated from 27.9% (39/140) of examined samples across dairy supply nodes and infant stool in Egypt, with equal peak prevalence in clinical mastitic milk (40.0%, 8/20) and infant fecal specimens (40.0%, 8/20). High rates of multidrug resistance (MDR; 66.7%, 26/39) were observed, with all isolates (100%) confirmed as extended-spectrum β-lactamase (ESBL) producers. Colistin (CT) and azithromycin resistance reached 46.2% (18/39) overall, peaking in commercial milk (69.2% for CT). Molecular screening revealed high frequencies of the mobile colistin resistance gene mcr-1 (41.0%, 16/39) and the ESBL determinant blaCTX-M (66.7%, 26/39), alongside fimbrial adhesion (mrkA, 76.9%) and iron acquisition (kfuBC, 82.1%) loci. All isolates (100%) exhibited strong biofilm-forming capacity (mean OD₅₇₀ = 0.63 ± 0.016), while toxin-antitoxin (TA) modules (symE, ccdB, relE) correlated positively with MDR profiles (p = 0.024). Bacterial virulence gene burden showed strong statistical associations with host tissue damage and inflammatory markers, including elevated milk lactate dehydrogenase (LDH; r = 0.918), haptoglobin (r = 0.954), and neonatal fecal calprotectin (FC; 83.34 ± 21.13 μg/g vs. 22.4 ± 2.0 μg/g in controls, p = 0.02). Crucially, milk-based cell-free metabolic supernatant (CFS) from Lactobacillus acidophilus demonstrated superior antagonistic activity compared to broth-based CFS (inhibition zones up to 40 mm vs. 12 mm against CT-resistant isolates). In silico molecular docking predicted strong binding affinities of putative L. acidophilus bioactive peptides (Acidocin 8912 score: −108.07 kcal/mol; Acidocin B score: 80.63 kcal/mol) to K. oxytoca outer membrane porin CymA and transport targets. These findings highlight the co-occurrence of resilient K. oxytoca lineages across food and pediatric matrices and demonstrate the therapeutic potential of probiotic-derived CFS as a non-antibiotic control strategy within a One Health framework.