Antimicrobial resistance is a major concern in the dairy industry. This study investigated the prevalence, antimicrobial resistance phenotypes, and genome sequencing of Gram-negative bacteria isolated from clinical (n = 350) and subclinical (n = 95) bovine mastitis, and raw unpasteurized milk (n = 125). Klebsiella pneumoniae, Aeromonas hydrophila, Enterobacter cloacae (100% each), Escherichia coli (87.78%), and Proteus mirabilis (69.7%) were the most prevalent multidrug-resistant (MDR) species. Extensive drug-resistance (XDR) phenotype was found in P. mirabilis (30.30%) and E. coli (3.33%) isolates. Ten isolates (four E. coli, three Klebsiella species and three P. mirabilis) that displayed the highest multiple antibiotic resistance (MAR) indices (0.54–0.83), were exposed to whole-genome sequencing (WGS). Two multilocus sequence types (MLST): ST2165 and ST7624 were identified among the sequenced E. coli isolates. Three E. coli isolates (two from clinical mastitis and one from raw milk) belonging to ST2165 showed similar profile of plasmid replicon types: IncFIA, IncFIB, IncFII, and IncQ1 with an exception to an isolate that contained IncR, whereas E. coli ST7624 showed a different plasmid profile including IncHI2, IncHI2A, IncI1α, and IncFII replicon types. ResFinder findings revealed the presence of plasmid-mediated colistin mcr-10 and fosfomycin fosA5 resistance genes in a K. pneumoniae (K1) isolate from bovine milk. Sequence analysis of the reconstructed mcr-10 plasmid from WGS of K1 isolate, showed that mcr-10 gene was bracketed by xerC and insertion sequence IS26 on an IncFIB plasmid. Phylogenetic analysis revealed that K1 isolate existed in a clade including mcr-10-harboring isolates from human and environment with different STs and countries [United Kingdom (ST788), Australia (ST323), Malawi (ST2144), Myanmar (ST705), and Laos (ST2355)]. This study reports the first emergence of K. pneumoniae co-harboring mcr-10 and fosA5 genes from bovine milk in the Middle East, which constitutes a public health threat and heralds the penetration of the last-resort antibiotics. Hence, prudent use of antibiotics in both humans and animals and antimicrobial surveillance plans are urgently required.

Bovine respiratory disease (BRD) is a complex syndrome associated with high mortality in young calves and causes severe economic losses in the cattle industry worldwide. The current study investigated the prevalence and molecular characterization of common bacterial pathogens associated with respiratory symptoms in young calves from Sadat City, one of the largest industrial cities in Menoufiya Governorate, Egypt. In between December 2020 and March 2021, 200 mixed-breed young calves of 6–12 months were examined clinically. Of them, sixty (30%) calves showed signs of respiratory manifestations, such as coughing, serous to mucopurulent nasal discharges, fever, and abnormal lung sound. Deep nasal (Nasopharyngeal) swabs were collected from the affected calves for bacteriological investigation. Phenotypic characterization and identification revealed Mycoplasma bovis, Mycoplasma bovigenitalium, Pasteurella multocida, and Staphylococcus aureus in 8.33%, 5%, 5%, and 5% of the tested samples, respectively. The PCR technique using species-specific primer sets successfully amplified the target bacterial DNA in all culture-positive samples, confirming the identity of the isolated bacterial species. Partial gene sequencing of 16S rRNA gene of M. bovigenitalium, P. multocida, and S. aureus, and mb-mp 81 gene of M. bovis revealed high nucleotide similarity and genetic relationship with respective bacterial species reported from Egypt and around the world, suggesting transmission of these bacterial species between animal host species and localities. Our study highlights the four important bacterial strains associated with respiratory disorders in calves and suggests the possible spread of these bacterial pathogens across animal species and different geographic locations. Further studies using WGS and a large number of isolates are required to investigate the realistic lineage of Egyptian isolates and globally.

An outbreak of brucellosis in a mixed dairy farm with 508 animals (370 cows, 120 sheep, and 18 camels) at Fayoum governorate, North Upper Egypt, was investigated. A storm of abortion and several cases of retained placenta were reported among cows and ewes in April 2020. Serodiagnosis of brucellosis was done using Rose Bengal Test (RBT) and Buffered Acidified Plate Antigen Test (BAPAT). The Milk Ring Test (MRT) was applied to the milk of seropositive animals. A total of 89 samples were used for isolation of Brucella and isolates were confirmed using Abortus, Melitensis, Ovis, Suis-PCR (AMOS-PCR). Test and slaughter strategy was applied to eradicate brucellosis from the farm based on RBT every month until three successive negative tests were obtained. Results showed that the seroprevalences of brucellosis based on RBT and BAPAT were 9.5%, 35%, and 50% in cattle, sheep, and camels, respectively. Despite 50% of male camels being seropositive, no clinical signs have been reported. The MRT identified fewer positive cases than BAPAT and RBT, thus, it cannot be used alone to eliminate the infection from the farm. A total of 31 Brucella isolates were recovered from cows and sheep on the farm. All isolates were confirmed as Brucella melitensis bv3 based on bacteriological examination and Brucella AMOS-PCR confirmed all isolates as Brucella melitensis. No positive reactors at the 6th, 7th, 8th, 9th, and 10th examinations were reported after the implementation of the test and slaughter strategy. In conclusion, extensive animal farming and mixed breeding are potential risk factors for interspecies transmission of brucellosis. Additionally, the test and slaughter strategy could be helpful to release the herd out of quarantine, however, application of biosecurity practices and fair compensation policy for owners should be implemented.

Keywordsklebsiella species are one of the key issues that have been steadily rising in intensive poultry production, causing great economic losses. The main cause of this uncomfortable condition is the increasing resistance of bacteria to antibiotics and has food safety risks because it can act as a source of contamination for chicken meat and eggs. In the present study, we aim to isolation and identification of klebsiella spp. and Molecular screening of antibiotics resistance genes that present in isolated Klebsiella spp. Twenty-Eight klebsiella species obtained from 50 healthy chickens differentiated into 21/28 (75%) Klebsiella pneumoniae and 7/28 (25%) Klebsiella oxytoca. Antimicrobial sensitivity testing against 12 commonly used antibiotics in chicken farms revealed that klebsiella species were fully resistant to oxytetracycline (100%) and penicillin (100%) and trimethoprim (100%) and moderate resistance to cefadroxil, doxycycline, amoxicillin/clavulanic acid, chloramphenicol, cefotaxime and lowest resistance to meropenem and klebsiella species extremely susceptible to amikacin (100%), tobramycin (100%) and norfloxacin (100%). A total of 12 klebsiella species differentiated into 9 klebsiella pneumoniae and 3 klebsiella oxytoca screened to find ESBL coding gene in the klebsiella species. The isolates were found to have bla SHV (100%), bla TEM (91.7%), and bla CTX-M (83.4%).

Frozen fish are exposed to many risks of contamination during long chain of catching, transportation, dressing and freezing from different sources. Till they reach to consumers which make it harmful or unfit for human consumption. A total of 100 random samples of frozen Saurus and Mackerel (50 of each) were collected from different fish markets at El -Menofia governorate. They were subjected to bacteriological examination for isolation of Psychotropic and Pseudomonas species. The bacteriological examination revealed that the mean values of total Psychotropic count in the examined samples of Saurus were1.07×10 7 ±2.82×10 6 and 1.14×10 7 ±3.2×10 6 in Mackerel. The incidence of Pseudomonas species was 40%and50% of the examined samples of Saurus and Mackerel, respectively. The incidence of identified Pseudomonas species isolated from the examined samples of frozen fish were Ps. aeruginosa, Ps. dimenuta, Ps. fluorescence, Ps. putida and Ps. fragi. The Pseudomonas species were resistant to chloramphenicol and nalidixic acid. In contrast they were sensitive to gentamycin except Ps.