Major Gram-negative bacterial causes isolated from apparent Healthy and diarrheic foals in Egypt, prevalence, identification and antibiotic susceptibility profiles

Diarrhea is one of the most significant illnesses affecting young foals and may be manifested in more than half of foals up to 3 months of age. Numerous infectious causes have been involved but bacterial pathogens are concerned. This study aimed to assess and identify the prime Gram-negative bacteriological etiology of Arabian horse foals’ diarrhea in Egypt and to designate the antibiotic susceptibility profiles of the isolated microorganisms.  Rectal swabs and internal organs were obtained from 216 foals (89 diarrheic and 127 apparently healthy), aged 1 week to 1 year old, reared in Arabian horse farms in Great Cairo, Egypt during a period extended from March 2022 to December 2022. Conventional bacteriological examination was performed using selective media persuaded by routine and advanced biochemical tests. The isolation displayed 648 bacterial isolates; 452 (68.6%) were Gram-negative with the mixed isolation representing about 28%. Escherichia coli constituted the most prevalent; 110 / 452 (24.34%); 65 in apparently healthy foals and 45 among diarrheic ones. The second was Klebsiella pneumoniae (72, 15.92%) at which 50, (15.67%) were found to expose the highest causative agent recovered from diarrheic foals. Imipenem, quinolones, and trimethoprim/sulfamethoxazole were the most effective versus Gram-negative species isolated from diarrheic foals while ampicillin and tetracycline were of no effect. The phenotypic antibiotic susceptibility test revealed a picture of multidrug resistance (MDR) as 111 isolates (28.71%) showed resistance to three or more antibiotics belonging to different groups. The results demonstrated that Klebsiella pneumonia was the highest MDR species (54.16%) followed by Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli isolates by 43.9, 32.1 and 30.9 % respectively. In conclusion, Gram-negative bacteria constituted the major causative agents of diarrhea in the Arabian foals. The most effective antibacterial drugs were imipenem, quinolones, and trimethoprim/ sulfamethoxazole. The issue of the existence of multidrug resistance isolates should be considered for proper therapy of foal diarrhea

Major Gram-negative bacterial causes isolated from apparent Healthy and diarrheic foals in Egypt, prevalence, identification and antibiotic susceptibility profiles

Diarrhea is one of the most significant illnesses affecting young foals and may be manifested in more than half of foals up to 3 months of age. Numerous infectious causes have been involved but bacterial pathogens are concerned. This study aimed to assess and identify the prime Gram-negative bacteriological etiology of Arabian horse foals’ diarrhea in Egypt and to designate the antibiotic susceptibility profiles of the isolated microorganisms.  Rectal swabs and internal organs were obtained from 216 foals (89 diarrheic and 127 apparently healthy), aged 1 week to 1 year old, reared in Arabian horse farms in Great Cairo, Egypt during a period extended from March 2022 to December 2022. Conventional bacteriological examination was performed using selective media persuaded by routine and advanced biochemical tests. The isolation displayed 648 bacterial isolates; 452 (68.6%) were Gram-negative with the mixed isolation representing about 28%. Escherichia coli constituted the most prevalent; 110 / 452 (24.34%); 65 in apparently healthy foals and 45 among diarrheic ones. The second was Klebsiella pneumoniae (72, 15.92%) at which 50, (15.67%) were found to expose the highest causative agent recovered from diarrheic foals. Imipenem, quinolones, and trimethoprim/sulfamethoxazole were the most effective versus Gram-negative species isolated from diarrheic foals while ampicillin and tetracycline were of no effect. The phenotypic antibiotic susceptibility test revealed a picture of multidrug resistance (MDR) as 111 isolates (28.71%) showed resistance to three or more antibiotics belonging to different groups. The results demonstrated that Klebsiella pneumonia was the highest MDR species (54.16%) followed by Pseudomonas aeruginosa, Salmonella enterica, and Escherichia coli isolates by 43.9, 32.1 and 30.9 % respectively. In conclusion, Gram-negative bacteria constituted the major causative agents of diarrhea in the Arabian foals. The most effective antibacterial drugs were imipenem, quinolones, and trimethoprim/ sulfamethoxazole. The issue of the existence of multidrug resistance isolates should be considered for proper therapy of foal diarrhea

Multidrug-resistant strains of Mycobacterium complex species in Egyptian farm animals, veterinarians, and farm and abattoir workers

Background and Aim: Mycobacterium tuberculosis complex (MTBC) is a group of mycobacteria that are important human pathogens. Mycobacterium tuberculosis and Mycobacterium bovis cause serious chronic life-threatening disease and also significant economic losses in both production and remedication. Recently, emergence of multidrug-resistant tuberculosis (MDR-TB) complex has generated global recognition of the need for rapid and sensitive diagnosis and development of new treatments. The current study illustrates the isolation/identification of MTBC strains in specimens obtained from cows and humans by conventional and real-time polymerase chain reaction (RT-PCR) techniques. Further, the study assesses sensitivity to antituberculosis drugs in isolated MDR strains. Materials and Methods: A total of 1464 samples from cattle (1285 raw milk and 179 lymph node), and 149 human sputum samples, were collected from farms and abattoirs in Delta Egypt. Conventional methods (culture and Ziehl–Neelsen staining) were implemented as were RT-PCR using MTBC universal DNA. The effect of some antituberculosis drugs on obtained isolates was assayed using drug susceptibility proportion and qualitative suspension techniques. Results: The MBTC detection rate using the culture method was higher than for Ziehl–Neelsen staining; raw cow milk (2.56 vs. 1.63%), lymph nodes (51.59 vs. 48.04%), and human sputum (5.36 vs. 4.02%). A total of 135 isolates were obtained. Application of RT-PCR detected 138 isolates from the same set of samples. MBTC isolates were resistant to first-line antituberculosis drugs, such as pyrazinamide, isoniazid, rifampicin, and ethambutol by 78.5, 59.3, 40.7, and 31.8%, respectively, and could be highly resistant to kanamycin (82.3%) and amikacin (80.7%). However, isolates remained sensitive to ciprofloxacin (71.1%) and clarithromycin (73.3%) as second-line drugs. Conclusion: There is a growing risk for isolation of MDR-TB from raw milk and lymph nodes of field tuberculin positive cattle as well as sputum of veterinarians and workers existed in farms and abattoirs. PCR-based techniques have become the gold standard for the identification of mycobacterial species, showing high efficiency compared to bacteriological and microscopic examination. Application of the first- and second-line antituberculosis drugs in combination could counter the MDR-TB concern once infections are identified

Advanced molecular characterization of enteropathogenic Escherichia coli isolated from diarrheic camel neonates in Egypt

Background and Aim: Camels are important livestock in Egypt on cultural and economic bases, but studies of etiological agents of camelid diseases are limited. The enteropathogen Escherichia coli is a cause of broad spectrum gastrointestinal infections among humans and animals, especially in developing countries. Severe infections can lead to death. The current study aimed to identify pathogenic E. coli strains that cause diarrhea in camel calves and characterize their virulence and drug resistance at a molecular level. Materials and Methods: Seventy fecal samples were collected from diarrheic neonatal camel calves in Giza Governorate during 2018-2019. Samples were cultured on a selective medium for E. coli, and positive colonies were confirmed biochemically, serotyped, and tested for antibiotic susceptibility. E. coli isolates were further confirmed through detection of the housekeeping gene, yaiO, and examined for the presence of virulence genes; traT and fimH and for genes responsible for antibiotic resistance, ampC, aadB, and mphA. The isolates in the important isolated serotype, E. coli O26, were examined for toxigenic genes and sequenced. Results: The bacteriological and biochemical examination identified 12 E. coli isolates from 70 fecal samples (17.1%). Serotyping of these isolates showed four types: O26, four isolates, 33.3%; O103, O111, three isolates each, 25%; and O45, two isolates, 16.7%. The isolates showed resistance to vancomycin (75%) and ampicillin (66.6%), but were highly susceptible to ciprofloxacin, norfloxacin, and tetracycline (100%). The structural gene, yaiO (115 bp), was amplified from all 12 E. coli isolates and traT and fimH genes were amplified from 10 and 8 isolates, respectively. Antibiotic resistance genes, ampC, mphA, and aadB, were harbored in 9 (75%), 8 (66.6%), and 5 (41.7%), respectively. Seven isolates (58.3%) were MDR. Real-time-polymerase chain reaction of the O26 isolates identified one isolate harboring vt1, two with vt2, and one isolate with neither gene. Sequencing of the isolates revealed similarities to E. coli O157 strains. Conclusion: Camels and other livestock suffer various diseases, including diarrhea often caused by microbial pathogens. Enteropathogenic E. coli serotypes were isolated from diarrheic neonatal camel calves. These isolates exhibited virulence and multiple drug resistance genes

Antigenic evaluation of extracted fimbrial protein obtained from pathogenic Escherichia coli isolated from diarrheic camel neonates

Enteropathogenic Escherichia coli (EPEC) were considered prime reason of diarrhea among neonatal livestock animals in developing countries and could be of public health importance via contaminated milk and meat.  Continual attempts were conducted to combat this illness using various antigenic determinants. This study was performed on four E. coli serotypes O26, O45, O103 and O111, which were previously recovered from diarrheic camel calves in Giza, Egypt. Extraction of fimbrial proteins was carried out through dialysis then evaluation of their immunogenic activity was preceded. SDS-PAGE electrophoresis was performed on crude extracted fimbrial proteins and revealed single band for each isolate ranged from 22 to 33kDa. Immunobloting was implemented on the extracted crude fimbrial proteins against E.coli O26 antisera formerly prepared in rabbit.These findings suggested that the fimbrial proteins are of immunogenicity importance and can serve as a protective passive vaccine antigen in prevention of diarrhea caused by EPEC and ETEC infection in camel calves