Shiga toxin-producing Escherichia coli (STEC) is a pathotype of E. coli that causes enteric and systemic diseases ranging from diarrhoea to severe hemorrhagic colitis (HC) and hemolytic uremic syndrome (HUS). The emergence of multidrug-resistant (MDR) STEC from cattle sources has increased public health risk and limited treatment options. The prevalence of STEC was investigated in 200 raw food samples (milk and beef samples) and 200 diarrheic samples (cattle and human samples) in a matched region. The presence of stx genes (stx1 and stx2), carbapenemase-encoding genes (blaVIM, blaNDM-1, and blaIMP), and extended-spectrum β-lactamase (ESBL)-encoding genes (blaTEM group, blaCTX-M1 group, and blaOXA-1 group) was screened by polymerase chain reaction (PCR). Antibiogram and Enterobacterial repetitive intergenic consensus (ERIC)-PCR were also conducted. STEC isolates were identified in 6.5% (13/200) of food samples [6% (6/100) of milk and 7% (7/100) of beef samples] and in 11% (22/200) of diarrheic cases [12% (12/100) of cattle and 10% (10/100) of human samples]. We found that O26 (4.5%, 18/400) and O111 (1.5%, 6/400) were the most prevalent STEC serovars and were found more commonly in diarrheic samples. STEC strains with both stx genes, stx2 only, and stx1 only genotypes were present in 62.9% (22/35), 20% (7/35), and 17.1% (6/35) of isolates, respectively. Carbapenemase-producing STEC (CP STEC) isolates were found in 1.8% (7/400) of samples [0.5% (1/200) of foods and 3% (6/200) of diarrheic cases]. The blaVIM gene was detected in all CP STEC isolates, and one human isolate carried the blaNDM-1 gene. ESBL-producing STEC strains were detected in 4.3% (17/400) of samples [1.5% (3/200) of food samples and 7% (14/200) of diarrheic cases]. The blaTEM, blaCTX-M1, and blaOXA-1 genes were detected in 42.9% (15/35), 28.6% (10/35), and 2.9% (1/35) of STEC isolates, respectively. Approximately half (51.4%, 18/35) of STEC isolates were MDR STEC; all CP STEC and ESBL-producing STEC were also MDR STEC. The highest antimicrobial resistance rates were found against nalidixic acid (51.4%) and ampicillin (48.6%), whereas the lowest rates were reported against gentamicin (5.7%) and ciprofloxacin (11.4%). MDR STEC strains were 5.3 times more likely to be found in diarrheic cases than in foods (P = 0.009, 95% CI 1.5–18.7). ERIC-PCR was used for genotyping STEC isolates into 27 different ERIC-types (ETs) with a discrimination index of 0.979. Five ETs showed clusters of 2–4 identical isolates that shared the same virulence and antibiotic resistance genetic profile. Human isolates matched food isolates in two of these ET clusters (the O26 CP STEC cluster and the O111 STEC cluster), highlighting the potential cross-species zoonotic transmission of these pathogens and/or their genes in the study region. This is the first detection of CP STEC in milk and diarrheic cattle in Egypt.
Dermatophytosis is considered as one of the most important fungal skin disease affecting both humans and animals otherwise its importance as zoonotic disease. Treatment of ringworm in cattle is expensive and time consuming especially on herd level. Vaccination can be a giant step forward to prevent the occurrence of mycozoonosis between the affected animals with ringworm and susceptible man. Fifty skins scraping samples were collected from animals (cattle, sheep and calves) suffering from skin lesions suspected to be ringworm. Then these collected samples were subjected to mycological examination and identification. T. verrucosum is the main causative agent of collected skin scrapings. Two types of vaccines were prepared from this strain (Culture filtrate and formalin inactivated vaccines). Rabbit model was used for challenge testing of the vaccines. The rabbits were separated into four groups as follow: Each group contained 3 rabbits each one was injected subcutaneously with 1ml of culture filtrate with adjuvant twice for seven days intervals. Group No. 2 rabbits each one was injected intramuscularly with 1 mL formalin-inactivated with adjuvant. The 3rd group is control positive and 4th one is control negative. Vaccination programme is done via injection twice with one week interval and the challenge by scarification of the skin with 0.2 mL of 5×10 7 fungal elements/mL. For 3 days. Humeral immune response was assessed by measuring anti-T. verrucosum antibodies using ELISA technique (direct method). By using statistical analysis of ANOVA, the mean optical density of anti-T. verrucosum specific IgG level in rabbits artificially immunized with culture filtrate with adjuvant vaccine significantly increase from (1.97) post first vaccination to reach (2.43) post second vaccination and still high after challenge with the virulent T. verrucosum strain (2.32) when compared with control positive (1.57). The results revealed that both type of vaccines induce good humeral immune response in vaccinated animals. Culture filtrate vaccine used as a treatment in infected farm animals by two doses within 2 weeks and noted that healing occurred.
Although () is a highly significant pathogen, its source remains unclear. Many people consume chicken daily as a source of animal protein worldwide; thus, hygienic methods of supplying chickens for consumption are critical for public health. Therefore, our study examined the distribution of the (), , and virulence genes in strains in chicken meat and giblets (gizzards and livers) and the resistance of the strains to various antibiotics. Ninety chicken meat, gizzard and liver samples were obtained from a semi-automatic abattoir in Sadat City, Egypt, and were cultured and preliminarily analyzed using biochemical tests. The presence of the ,, and genotypes was tested for in samples positive for by multiplex polymerase chain reaction (Multiplex-PCR). The resistance of to various antimicrobial drugs was tested using the disc diffusion method. In total, 7 of the 90 chicken samples were positive for (7.78%); in 3/7 (42.85%) samples, the bacteria were found in the chicken liver, while the bacteria were found in the meat in 2/7 (28.57%) and in the gizzard in 2/7 (28.57%) samples. The total prevalence of both the and genes in the isolated strains was 100%, while the prevalence of the and genes was 57.1% and 42.9%, respectively. The resistance of to the antibiotics utilized in our study was 100% for streptomycin; 85.7% for amoxicillin and penicillin; 71.4% for oxytetracycline, nalidixic acid and ampicillin; 57.1% for sulfamethoxazole and erythromycin; and 42.9% for neomycin, chloramphenicol and norfloxacin. In conclusion, the chicken meat and giblets were tainted by, with a higher occurrence of the ,, and genotypes. Future investigations should investigate the resistance of to various antimicrobial agents in Egypt.
Mastitis is one of the most economic disease affecting dairy cows worldwide. Identification of mastitis pathogens still depends principally on culture and phenotypical method, which is a difficult and timeconsuming. Newly, microbiologists have focused their attention on the use of Mass Spectrometry (MS) for microbial identification, especially Matrix Assisted Laser Desorption Ionization Time-Of-Flight (MALDI-TOF). Therefore, this study was designated to evaluate the ability of MALDI-TOF to identify some contagious mastitis pathogens comparing with phenotypical methods such as API panels and VITEK 2 system. A total of one hundred twenty of Staphylococcus aureus (S. aureus), Coagulase Negative Staphylococci (CNS) and Streptococcus agalactiae (Strept. agalactiae) strains isolated from milk of cows affected by clinical and subclinical mastitis were used in the study. According to the results, ~95% of S. aureus, 100% of CNS and Strept. agalactiae were correctly identified by MALDI TOF MS. All S. aureus isolates were then confirmed by a nuc-based PCR technique. While ~92% of S. aureus, 87% of Strept. agalactiae and 76% of CNS were identified by VITEK 2 system. Moreover, ~89% of S. aureus, 80% of Strept. agalactiae and 72% of CNS were identified by API system. In brief, the results demonstrated that MALDI-TOF is a fast and truthful technique which has the capability to replace conventional identification of several bacterial strains usually isolated in clinical laboratories of microbiology. Therefore, it is recomended that MALDI-TOF MS technology can be regularly used in veterinary laboratories for identification of different species of bacteria, particularly when failure of phenotypic methods forces clinical microbiologists.
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