A total of 150 random samples were collected from different supermarkets and retailers of differe... more A total of 150 random samples were collected from different supermarkets and retailers of different sanitation levels at Gharbeia governorate, Egypt as follows 70 samples of minced meat, 40 samples of sausage and 40 samples of beef burger. The aforementioned samples were subjected to bacteriological and serological applications to assess the prevalence of S. Enteritidis. The obtained results revealed that the incidence of S. Enteritidis in the examined samples of minced meat, sausage and beef burger were 1/70 (1.4%), 1/40(2.5%) and 0/40(0%) respectively. The isolates were submitted for serological analysis and revealed that Salmonella Enteritidis O 1,9,12 ad monophasic H:g, m. The antibiogram sensitivity test was applied upon the two isolates of S. Enteritidis and revealed that they are sensitive to chloramphinicol, amoxicillin, levofloxacin, ciprofloxacin, enrofloxacin and gentamycin but they were resistant to oxytetracycline. The virulence genes of S. Enteritidis isolates were determined by using multiple PCR technique for the two serologically detected Salmonella Enteritidis by using the following genes inv A, sef A, sop B and bcf C at 284bp, 310bp, 517bp, 467bp, respectively.
The present study was performed on 250 random samples of fresh meat and meat products. Beef burge... more The present study was performed on 250 random samples of fresh meat and meat products. Beef burger, kofta, minced meat and sausage (50 for each) were collected from different shops (25gm of each sample) at Kaliobia Governorate, Egypt, to detect the prevalence of some toxigenic food-borne bacteria, beside the phenotypic characterization and detection of some virulence genes. Bacteriological examination of the collected samples resulted in an isolation of Staph. aureus isolates (41/16.4%), E. coli (25 /10.0%), B. cereus (21/8.4%) and Salmonella (3/1.2%). The antibiotic sensitivity tests for the isolated strains showed multiple antibiotic resistances (ampicillin, methicillin, oxytetracycline, amoxicillin, streptomycin, erythromycin, doxycycline and cefotaxime). Therefore, E. coli; Staph. aureus and B. cereus strains especially antibiotic resistances ones are meat-borne pathogens of public health important Antibiotic resistant Food borne pathogen Meat products
In this study the incidence of Bacillus cereus (B. cereus) and antibiotic sensitivity test was un... more In this study the incidence of Bacillus cereus (B. cereus) and antibiotic sensitivity test was undertaken. A total of one hundred random samples of meat products of rice kofta, kobeba, chicken pane and chicken nuggets (25 of each) were collected from different supermarkets at different times in Menofiea and Kalyobia governorates, Egypt and tested for occurrence of Bacillus group. Bacillus cereus were detected in 24%, 12%, 20% and 10% of kobiba-shami, rice kofta, chicken pane and chicken nuggets, respectively. Bacillus mycoids failed to be isolated from kobiba-shami and isolated from 4%, 16% and 4% of rice kofta, chicken pane and chicken nuggets, respectively. Also, B. thuringenesis failed to be isolated from kobibashami and rice kofta while it isolated from 8% of both chicken pane and chicken nuggets. Fourteen strains of B. cereus were examined for antimicrobial susceptibility testing. The foremost common drug resistance was to penicillin, amoxicillin and amoxicillin + clavulanic (100% for each). On the other hand, B. cereus was completely prone to vancomycin and gentamycin (100% for each).Using PCR, all tested B. cereus isolates harbored bla gene B. cereus bla resistance gene Chicken and Meat products
The present study was carried out on 120 examined cultured Nile tilapia, Oreochromis niloticus (O... more The present study was carried out on 120 examined cultured Nile tilapia, Oreochromis niloticus (O. niloticus) (80 diseased fish and 40 apparently healthy) were collected alive from private fish farms at Kafr El Sheikh Governorates from April to October 2018. Grossly, examined diseased O. niloticus revealed eroded fins, redness on the skin, congested protruded anal opening and sometimes darkness on skin, ulcers with different sizes, and eye turbidity especially in fish naturally infected with Aspergillus, Fusarium and Pencillium. Mycological examination revealed isolation of 7 Genera of fungi from total incidence of 300 fungal isolates; 240 fungal isolates from diseased O. niloticus and 60 isolates from apparently healthy one. Aspergillus flavus and Aspergillus niger were the most predominant fungi isolated from either apparently healthy or even diseased one mainly from internal organs. Molecular identification was performed using inter-transcriped spacer (ITS) gene for A. flavus and A. niger. Developed PCR assay specific primer detect A. flavus and A. niger showing clear bands at 595 and 600 bp molecular weight, respectively.
Candidiasis is an occasional opportunistic yeast disease of importance in poultry has been report... more Candidiasis is an occasional opportunistic yeast disease of importance in poultry has been reported to be in intestinal infection. The present study was directed mainly to throw lights on the isolation of C. albicans species from chicken and the identification of the isolates by phenotypic and genotypic methods. A total of 100 swabs samples collected from a crop of chicken from different localities in Cairo and Giza Governorates. The samples were investigated for mycological examination. The most frequent yeast species isolated from such swabs were Candida, Saccharomyces, Torulopsis and Rhodotorula species. The incidence of Candida species was (n=36/60, 60%), and the incidence of Candida albicans was (n=8/36, 22.2%) from crop swab. Early identification of yeast to the genus and species level was necessary for effective antifungal therapy and can also facilitate control such pathogens infections.
Campylobacteriosis incidence and proportion of Campylobacter strains resistant to antibiotic have... more Campylobacteriosis incidence and proportion of Campylobacter strains resistant to antibiotic have been increased worldwide in the last decades. Campylobacteriosis is considered as one of the major important zoonotic gastrointestinal diseases around the world. The aim of this study was isolation and biochemical characterization of Campylobacter from poultry. Campylobacter isolation have been done on MCCDA media, motility has been detected by phase contrast microscope, morphology has been detected by Gram stain, and confirmed by MALDI-TOF MS and molecular confirmation by using cPCR by amplification of 16S rRNA gene. A total of 102 isolates of Campylobacter were isolated from farm and small backyards located in different sources in El-Kalyobia , El-Monofia and El-sharkia Governorates by conventional bacteriological methods from which 40 were C. jejuni and 62 were C. coli. All isolates have been confirmed as Campylobacter by MALDI-TOF MS and detection of 16S rRNA by cPCR. PCR is a useful molecular tool for identification and confirmation of Campylobacter. It is rapid, sensitive, and specific than the culture methods, but the only disadvantage is that is expensive.
Escherichia coli (E. coli) is the most significant cause of neonatal diarrhea in ruminants. The c... more Escherichia coli (E. coli) is the most significant cause of neonatal diarrhea in ruminants. The current study was designed to monitor the prevalence of multi-drug (MDR) resistant E. coli in diarrheic ruminants in Egypt. Rectal swab samples (n=150) were collected between November 2018 and April 2019 from diarrheic calves (n=35), lambs (n=35) and goat kids (n=80) up to 3 months from Gimmeza animal production researches station, Agriculture Research Centre (ARC), Egypt. Samples were submitted for isolation and identification of E. coli by conventional culture methods. From the examined samples , 82 (54.67%) were positive for E. coli. Among the E. coli isolates, 48 were identified as pathogenic E. coli by cultivation on Congo Red Agar and were submitted for antibiogram. Sensitivity tests revealed that 10/48 (20.83%) isolates were MDR. The 10 MDR E. coli isolates were serologically identified as O157:H7 (n=4; two isolated from calves and two from goat kids), O125 (n=3; two isolated from calves and one from lambs), O44 (n=3; two isolated from goat kids and one from lambs).
This study aimed to detect C. burnetii antibodies in milk and serum of dairy cattle using PCR com... more This study aimed to detect C. burnetii antibodies in milk and serum of dairy cattle using PCR compared with other immunological techniques namely IFAT and ELISA. These samples were collected from apparently healthy cows (n=300) from Giza, Fayoum and Beni Suief Governorates. Real-time PCR for detection of C. burnetii DNA was performed on samples using two individual PCR assays with specific primers and probes to increase specificity of the result using Roche kits. IFAT revealed detection of 6% IgM antibodies in milk and 7.3% in sera. While IgG was 10.6% of milk and 24% in sera. The main value of positive samples by ELISA test were 25.7%; in serum 34% and in milk 17.3%. These results showed that sensitivity and specificity of the ELISA test and IFAT were highly comparable. The result of Real time PCR from 77 positive samples of C. burnetii was 45.4%. The highest results for real time PCR recorded 57.1% in Beni Suief followed by 44% in Giza and 33.3% in Fayoum. It was concluded that the apparently healthy dairy cows are an important reservoir of C. burnetii infection. Investigations on C. burnetii using PCR as well as serological surveys of animals are important methods for diagnosis and control of Q-fever. Awareness is needed for animal owners, veterinarians, physicians and authorities.
The present study was conducted on 250 diseased Nile tilapia (Oreochromis niloticus) of various s... more The present study was conducted on 250 diseased Nile tilapia (Oreochromis niloticus) of various sizes collected from different fish markets at Kaliobia Governorate, Egypt, during the period from May 2017 to January 2019 for inspection of Aeromonas and Pseudomonas strains. Samples were collected from apparently pathognomonic lesions in muscle, liver, kidney, intestine and spleen for bacteriological examination. The results revealed that, 161 Aeromonas and Pseudomonas species; 118 Aeromonas (73.3%) and 43 Pseudomonas species (26.7%) were isolated mostly from 50 muscle lesion samples followed by 41 liver, 35 kidney, 32 intestine and 3 spleen lesion samples. Aeromonas strains were highly sensitive to meropenem followed by ciprofloxacin, norfloxacin, gentamycin and florphenicol. Meanwhile, they were highly resistant for ampicillin and methicillin followed by oxacillin, penicillin-G, amoxicillin, cefotaxime, oxytetracycline and streptomycin. In addition, Pseudomonas strains were highly sensitive to meropenem followed by gentamycin, norfloxacin, ciprofloxacin and florphenicol. In contrast, they were highly resistant for ampicillin, methicillin and penicillin-G followed by amoxicillin, oxacillin and cefotaxime.
Enterococcus species are considered as a major etiological agent of nosocomial infections, they a... more Enterococcus species are considered as a major etiological agent of nosocomial infections, they are commonly isolated from different sources of food. So, this study was conducted for detection of Enterococcus spp. from fish (n=10), herbs (n=30), drinking water (n=20), dairy products (n=30), and meat products (n=20) samples from Cairo Governorate, Egypt. PCR was done for detection of some antibiotic resistant genes (mphC, norA, tetK, floR, vanA). The results revealed that from 110 food samples analyzed, 11.8% were positive for Enterococci. Moreover, E. faecalis was detected by percentage 53.8%. E. faecalis isolates were resistant to vancomycin (100%), erythromycin (57.1%), ciprofloxacin (14.3%), chloramphenicol (42.9%), tetracycline (57.1%), while none of the E. faecalis isolates were resistant to ampicillin, penicillin or norfloxacin. Genotypic characterization revealed that tetK and floR genes were present in the all 7 E. faecalis isolates. While, vanA gene was detected in 3 isolates, and mphC gene was detected only in 2 isolates. The results of our investigation indicated high levels of contamination with multi-resistant E. faecalis strains of serious concern with the isolation of strains resistant to vancomycin, considering that vancomycin is the alternative agent for patients who are intolerant to penicillin or who have Enterococci infections with high level resistance to penicillin.
The present study was conducted on 250 random samples of meat and meat products (minced meat, kof... more The present study was conducted on 250 random samples of meat and meat products (minced meat, kofta, beef burger and sausage, 50 for each) collected from small retails and different supermarkets at Kaliobia Governorate, Egypt for inspection of Lactobacillus (LAB) species and studying the bioactivities of them. The results revealed that 252 strains of LAB strains were recovered from 250 examined samples. P. acidilactici were the most isolated (102 = 40.5%) followed by L. plantarum (76=30.2%), L. acidophilus (38=15.1%); L. brevis (19=7.5%) and P. pentosaceus (17=6.7%). All of them produced bacteriocin and biosurfactant that inhibited the growth of tested pathogenic bacteria. In addition, most isolated strains had the ability for biofilm production which able to inhibit the biofilm formation of tested pathogenic strains. Moreover, the sequences obtained for 16S rRNA genes had accession numbers MK850930 and MK852180, MK852397 and MK852398, MK806485 and MK850564, MK871658 and MK871674, MK852683 and MK852691, and were 91% to 98% identical to the corresponding GenBank sequences.
Frozen fish are exposed to many risks of contamination during long chain of catching, transportat... more 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. fluorescence AB susceptibility
In this comparative study, five Brucella strains were morphologically, serologically, and biochem... more In this comparative study, five Brucella strains were morphologically, serologically, and biochemically identified using media different stains and antibiotics. Multiplex PCR was used to differentiate between the Brucella strains and Yersinia enterocolitica O: 9. five primer sets were designed to the most specific variable regions of the different Brucella species. The results of multiplex PCR were very successful and accurate in terms of characterization and typing of the Brucella vaccine, reference and wild Brucella strains from Yersinia which gave false positive while using tests based on anti-LPS antibodies but with PCR for DNA of Yersinia gave negative. The molecular typing of the Brucella strains by multiplex PCR had several advantages over the use of the conventional methods being very fast, precise, easier, more sensitive, and economic and could be applied on minimal sample preparation. The development of this PCR method is the first step toward the development of a novel kit for the molecular identification of Brucella strains from other Gram-negative bacteria.
The aim of this study was to isolate Salmonellae from 579 birds (348 chickens, 104 ducks, 30 turk... more The aim of this study was to isolate Salmonellae from 579 birds (348 chickens, 104 ducks, 30 turkeys, 50 quail, 30 pigeons and 17 geese) from 4 Egyptian Governorates. The Samples collected from internal organs (liver, cecum, spleen and heart) were examined bacteriologically and serologically. Sixty-three (10.9%) out of 579 birds were found positive while 516 (89.1%) birds were negative for Salmonella isolation. The number and percentage of positive chickens, ducks, turkeys, quails, pigeons and geese were 43 (12.4%), 10 (9.6%), 3 (10%), 5 (10%), 2 (6.7%) and 0 (0%) respectively. In this study,
A total of 150 random samples were collected from different supermarkets and retailers of differe... more A total of 150 random samples were collected from different supermarkets and retailers of different sanitation levels at Gharbeia governorate, Egypt as follows 70 samples of minced meat, 40 samples of sausage and 40 samples of beef burger. The aforementioned samples were subjected to bacteriological and serological applications to assess the prevalence of S. Enteritidis. The obtained results revealed that the incidence of S. Enteritidis in the examined samples of minced meat, sausage and beef burger were 1/70 (1.4%), 1/40(2.5%) and 0/40(0%) respectively. The isolates were submitted for serological analysis and revealed that Salmonella Enteritidis O 1,9,12 ad monophasic H:g, m. The antibiogram sensitivity test was applied upon the two isolates of S. Enteritidis and revealed that they are sensitive to chloramphinicol, amoxicillin, levofloxacin, ciprofloxacin, enrofloxacin and gentamycin but they were resistant to oxytetracycline. The virulence genes of S. Enteritidis isolates were determined by using multiple PCR technique for the two serologically detected Salmonella Enteritidis by using the following genes inv A, sef A, sop B and bcf C at 284bp, 310bp, 517bp, 467bp, respectively.
The present study was carried out to evaluate the antimicrobial activity of ethanol plant extract... more The present study was carried out to evaluate the antimicrobial activity of ethanol plant extract of rosemary (Rosmarinus officinalis) and / with five antimicrobial agents of different mechanisms (oxytetracycline Hcl, amoxicillin, cefquinome, sulphaquinoxaline and danofloxacin) against field strain of S. aureus. By using agar well diffusion method, the mean zone of inhibition (mm) of ethanol extract
The study was performed on 225 chicken (72 diseased and 153 freshly dead ones) from different 45 ... more The study was performed on 225 chicken (72 diseased and 153 freshly dead ones) from different 45 commercial broiler farms (1-30 days old) at Dakahlia Governorate for mycotic infection. Samples were taken from these chickens (lung; air sac; crop; liver and brain from each chicken) after clinical and postmortem examination for mycological examination. The results revealed that, fungi were isolated from 503 positive samples (44.7%); represented as 119 positive samples (10.6%) from diseased chickens and 384 positive samples (34.1 %) from freshly dead ones.651 fungal isolates (moulds and yeasts) were isolated from 1125 samples, where 219(33.6%) were isolated from lung samples followed by 199(30.6%); 119(18.3%); 91(14.0%) and 23(3.5%) from air sac, liver, crop and brain samples respectively. Moreover, A. fumigatus was the most isolated one 141(21.7%) followed by A. flavus 126(19.4%); A. niger 111(17.1%); Rhizopus spp. 67(10.3%); C. albicans 58(8.9%); Mucor spp. 42(6.5%); Penicillium spp. 32(4.9%); A. terreus 20(3.1%); Fusarium spp.19 (2.9%); C. krusei 13(2.0%); C. tropicalis 9(1.4%); A. candidus 9(1.4%) and finally A. ochraceus 4(0.6%). PCR using fungus-specific universal primer pairs (ITS1 and ITS4) was used for identification and genotypic characters of Aspergillus species (A. fumigatus; A. flavus and A. niger).They were able to successfully amplify the ITS1-5.8S rDNA region of all tested Aspergillus isolates, providing a single PCR product of about 600 bp for all tested isolates PCR using fungus-specific universal primer pairs (ITS and RPS) was used for identification and genotypic characters of C. providing a single PCR product of about 109 bp .Sequence of the internal transcribed spacer 1 reigon (ITS-1) of isolated A. flavus Gene Bank accession number for studied nucleotide sequence (Bankit 1867303) is KM 983253. The sequences obtained for ITS-1 region were more than 98% identical to the corresponding GenBank sequences (accession no. DQ467968.1; KC994648.1; LN482516.1 and KP689246.1). Sequence of the internal transcribed spacer 1 region (ITS-1) of isolated C. albicans. Gene Bank accession number for studied nucleotide sequence (Bankit 1867955) is KM 983254. The sequences obtained for ITS-1 region obtained were more than 95% identical to the corresponding GenBank (accession no. M87288.1; gb|S71769.1|S71769; emb (AL033396.1) and AP006852.1).
A total of (236 swabs) fecal samples from El-Menofiya and El-Kalubia Governorates, as sporadic ca... more A total of (236 swabs) fecal samples from El-Menofiya and El-Kalubia Governorates, as sporadic cases of were subjected to bacteriological, biochemical, serotyping, sensitivity testing and PCR detection of resistance genes for β-Lactames and Quinolones. Ten isolates of Salmonella species were identified, as 7 isolates (4.7%) from calves (Salmonella Typhimurium, Salmonella Enteritidis, Salmonella saintpaul, Salmonella Langeveld and Salmonella Havana), 2 isolates (3.6%) from lambs (Salmonella Typhimurium, Salmonella Bardo) and one isolate (3.3%) from goat-kids (Salmonella Enteritidis) Salmonella isolates sensitivity to β-lacatmes was 0.00% for ampicillin, penicillin G, piperacillin, cephalexin, cefoxitin, ampicillin-sulbactam, ceftazidime, 10% for ceftriaxone and 20% for amoxicillinclavulanic acid, increased to 60% and 100% for aztreonam, imipenem respectively. Susceptibility of isolates to quinolones were 10% for ciprofluxacin, 20% to nalidixic acid, and 100% for each of norfloxacin and levofloxacin. PCR study showed beta-lactamase encoding gene, bla (TEM-1), was identified in 90% and the extended-spectrum beta-lactamase, (CIT) in 20% while bla (SHV) and acc in 80% of isolates. The detection was 50%, 40%, and 0.00% for FOX, MOX, and bla (OXA-1) genes, respectively. Plasmid-mediated quinolone resistance, qnrB, qnrS were detected in 80% while aac(6')-Ib-cr, was only in 50% of the isolates. Results showed a high incidence of β-lacatmase than quinolones resistance genes and higher isolate susceptibility to quinolone than to β-lacatmes which indicates higher efficacy and validity of quinolones. Results indicates quinolones LEV and NOR in addition to βlacatmes imipenem are the drugs of choice for suspected salmonella cases.
Ten isolates of Salmonella species were obtained from sporadic cases of profuse diarrhea in new b... more Ten isolates of Salmonella species were obtained from sporadic cases of profuse diarrhea in new born ruminants from El-Menofiya and El-Kalubia Governorates in Egypt. These isolates were as follow 7 isolates from calves (Salmonella Typhimurium, Salmonella Enteritidis,two were Salmonella Saintpaul, two were Salmonella Langeveld and Salmonella Havana) ,two isolates from lambs (Salmonella Typhimurium, Salmonella Bardo) and one isolate from goat-kids(Salmonella Enteritidis). Salmonella isolates resistance to aminoglycosoides was 0.00% for Amikacin, 30% for tobramycin and 50% for streptomycin. Susceptibility of isolates to tetracycline was 50% for doxycycline. Results showed a high incidence of aminoglycosoides resistance gene aadB in 100% of the isolates while aadA2 genes in 40% of the isolates. Incidence of tetracycline resistance genes was 0.00% for tetA(B) and 80% for tetA(A). The difference between the results of this study and those from other regions in Egypt necessitate a complete survey overall the country to make a complete and clear map of salmonella servoars, their antibiotics susceptibility and in molecular characterization of resistance determinants in each region.
A total of 150 random samples were collected from different supermarkets and retailers of differe... more A total of 150 random samples were collected from different supermarkets and retailers of different sanitation levels at Gharbeia governorate, Egypt as follows 70 samples of minced meat, 40 samples of sausage and 40 samples of beef burger. The aforementioned samples were subjected to bacteriological and serological applications to assess the prevalence of S. Enteritidis. The obtained results revealed that the incidence of S. Enteritidis in the examined samples of minced meat, sausage and beef burger were 1/70 (1.4%), 1/40(2.5%) and 0/40(0%) respectively. The isolates were submitted for serological analysis and revealed that Salmonella Enteritidis O 1,9,12 ad monophasic H:g, m. The antibiogram sensitivity test was applied upon the two isolates of S. Enteritidis and revealed that they are sensitive to chloramphinicol, amoxicillin, levofloxacin, ciprofloxacin, enrofloxacin and gentamycin but they were resistant to oxytetracycline. The virulence genes of S. Enteritidis isolates were determined by using multiple PCR technique for the two serologically detected Salmonella Enteritidis by using the following genes inv A, sef A, sop B and bcf C at 284bp, 310bp, 517bp, 467bp, respectively.
The present study was performed on 250 random samples of fresh meat and meat products. Beef burge... more The present study was performed on 250 random samples of fresh meat and meat products. Beef burger, kofta, minced meat and sausage (50 for each) were collected from different shops (25gm of each sample) at Kaliobia Governorate, Egypt, to detect the prevalence of some toxigenic food-borne bacteria, beside the phenotypic characterization and detection of some virulence genes. Bacteriological examination of the collected samples resulted in an isolation of Staph. aureus isolates (41/16.4%), E. coli (25 /10.0%), B. cereus (21/8.4%) and Salmonella (3/1.2%). The antibiotic sensitivity tests for the isolated strains showed multiple antibiotic resistances (ampicillin, methicillin, oxytetracycline, amoxicillin, streptomycin, erythromycin, doxycycline and cefotaxime). Therefore, E. coli; Staph. aureus and B. cereus strains especially antibiotic resistances ones are meat-borne pathogens of public health important Antibiotic resistant Food borne pathogen Meat products
In this study the incidence of Bacillus cereus (B. cereus) and antibiotic sensitivity test was un... more In this study the incidence of Bacillus cereus (B. cereus) and antibiotic sensitivity test was undertaken. A total of one hundred random samples of meat products of rice kofta, kobeba, chicken pane and chicken nuggets (25 of each) were collected from different supermarkets at different times in Menofiea and Kalyobia governorates, Egypt and tested for occurrence of Bacillus group. Bacillus cereus were detected in 24%, 12%, 20% and 10% of kobiba-shami, rice kofta, chicken pane and chicken nuggets, respectively. Bacillus mycoids failed to be isolated from kobiba-shami and isolated from 4%, 16% and 4% of rice kofta, chicken pane and chicken nuggets, respectively. Also, B. thuringenesis failed to be isolated from kobibashami and rice kofta while it isolated from 8% of both chicken pane and chicken nuggets. Fourteen strains of B. cereus were examined for antimicrobial susceptibility testing. The foremost common drug resistance was to penicillin, amoxicillin and amoxicillin + clavulanic (100% for each). On the other hand, B. cereus was completely prone to vancomycin and gentamycin (100% for each).Using PCR, all tested B. cereus isolates harbored bla gene B. cereus bla resistance gene Chicken and Meat products
The present study was carried out on 120 examined cultured Nile tilapia, Oreochromis niloticus (O... more The present study was carried out on 120 examined cultured Nile tilapia, Oreochromis niloticus (O. niloticus) (80 diseased fish and 40 apparently healthy) were collected alive from private fish farms at Kafr El Sheikh Governorates from April to October 2018. Grossly, examined diseased O. niloticus revealed eroded fins, redness on the skin, congested protruded anal opening and sometimes darkness on skin, ulcers with different sizes, and eye turbidity especially in fish naturally infected with Aspergillus, Fusarium and Pencillium. Mycological examination revealed isolation of 7 Genera of fungi from total incidence of 300 fungal isolates; 240 fungal isolates from diseased O. niloticus and 60 isolates from apparently healthy one. Aspergillus flavus and Aspergillus niger were the most predominant fungi isolated from either apparently healthy or even diseased one mainly from internal organs. Molecular identification was performed using inter-transcriped spacer (ITS) gene for A. flavus and A. niger. Developed PCR assay specific primer detect A. flavus and A. niger showing clear bands at 595 and 600 bp molecular weight, respectively.
Candidiasis is an occasional opportunistic yeast disease of importance in poultry has been report... more Candidiasis is an occasional opportunistic yeast disease of importance in poultry has been reported to be in intestinal infection. The present study was directed mainly to throw lights on the isolation of C. albicans species from chicken and the identification of the isolates by phenotypic and genotypic methods. A total of 100 swabs samples collected from a crop of chicken from different localities in Cairo and Giza Governorates. The samples were investigated for mycological examination. The most frequent yeast species isolated from such swabs were Candida, Saccharomyces, Torulopsis and Rhodotorula species. The incidence of Candida species was (n=36/60, 60%), and the incidence of Candida albicans was (n=8/36, 22.2%) from crop swab. Early identification of yeast to the genus and species level was necessary for effective antifungal therapy and can also facilitate control such pathogens infections.
Campylobacteriosis incidence and proportion of Campylobacter strains resistant to antibiotic have... more Campylobacteriosis incidence and proportion of Campylobacter strains resistant to antibiotic have been increased worldwide in the last decades. Campylobacteriosis is considered as one of the major important zoonotic gastrointestinal diseases around the world. The aim of this study was isolation and biochemical characterization of Campylobacter from poultry. Campylobacter isolation have been done on MCCDA media, motility has been detected by phase contrast microscope, morphology has been detected by Gram stain, and confirmed by MALDI-TOF MS and molecular confirmation by using cPCR by amplification of 16S rRNA gene. A total of 102 isolates of Campylobacter were isolated from farm and small backyards located in different sources in El-Kalyobia , El-Monofia and El-sharkia Governorates by conventional bacteriological methods from which 40 were C. jejuni and 62 were C. coli. All isolates have been confirmed as Campylobacter by MALDI-TOF MS and detection of 16S rRNA by cPCR. PCR is a useful molecular tool for identification and confirmation of Campylobacter. It is rapid, sensitive, and specific than the culture methods, but the only disadvantage is that is expensive.
Escherichia coli (E. coli) is the most significant cause of neonatal diarrhea in ruminants. The c... more Escherichia coli (E. coli) is the most significant cause of neonatal diarrhea in ruminants. The current study was designed to monitor the prevalence of multi-drug (MDR) resistant E. coli in diarrheic ruminants in Egypt. Rectal swab samples (n=150) were collected between November 2018 and April 2019 from diarrheic calves (n=35), lambs (n=35) and goat kids (n=80) up to 3 months from Gimmeza animal production researches station, Agriculture Research Centre (ARC), Egypt. Samples were submitted for isolation and identification of E. coli by conventional culture methods. From the examined samples , 82 (54.67%) were positive for E. coli. Among the E. coli isolates, 48 were identified as pathogenic E. coli by cultivation on Congo Red Agar and were submitted for antibiogram. Sensitivity tests revealed that 10/48 (20.83%) isolates were MDR. The 10 MDR E. coli isolates were serologically identified as O157:H7 (n=4; two isolated from calves and two from goat kids), O125 (n=3; two isolated from calves and one from lambs), O44 (n=3; two isolated from goat kids and one from lambs).
This study aimed to detect C. burnetii antibodies in milk and serum of dairy cattle using PCR com... more This study aimed to detect C. burnetii antibodies in milk and serum of dairy cattle using PCR compared with other immunological techniques namely IFAT and ELISA. These samples were collected from apparently healthy cows (n=300) from Giza, Fayoum and Beni Suief Governorates. Real-time PCR for detection of C. burnetii DNA was performed on samples using two individual PCR assays with specific primers and probes to increase specificity of the result using Roche kits. IFAT revealed detection of 6% IgM antibodies in milk and 7.3% in sera. While IgG was 10.6% of milk and 24% in sera. The main value of positive samples by ELISA test were 25.7%; in serum 34% and in milk 17.3%. These results showed that sensitivity and specificity of the ELISA test and IFAT were highly comparable. The result of Real time PCR from 77 positive samples of C. burnetii was 45.4%. The highest results for real time PCR recorded 57.1% in Beni Suief followed by 44% in Giza and 33.3% in Fayoum. It was concluded that the apparently healthy dairy cows are an important reservoir of C. burnetii infection. Investigations on C. burnetii using PCR as well as serological surveys of animals are important methods for diagnosis and control of Q-fever. Awareness is needed for animal owners, veterinarians, physicians and authorities.
The present study was conducted on 250 diseased Nile tilapia (Oreochromis niloticus) of various s... more The present study was conducted on 250 diseased Nile tilapia (Oreochromis niloticus) of various sizes collected from different fish markets at Kaliobia Governorate, Egypt, during the period from May 2017 to January 2019 for inspection of Aeromonas and Pseudomonas strains. Samples were collected from apparently pathognomonic lesions in muscle, liver, kidney, intestine and spleen for bacteriological examination. The results revealed that, 161 Aeromonas and Pseudomonas species; 118 Aeromonas (73.3%) and 43 Pseudomonas species (26.7%) were isolated mostly from 50 muscle lesion samples followed by 41 liver, 35 kidney, 32 intestine and 3 spleen lesion samples. Aeromonas strains were highly sensitive to meropenem followed by ciprofloxacin, norfloxacin, gentamycin and florphenicol. Meanwhile, they were highly resistant for ampicillin and methicillin followed by oxacillin, penicillin-G, amoxicillin, cefotaxime, oxytetracycline and streptomycin. In addition, Pseudomonas strains were highly sensitive to meropenem followed by gentamycin, norfloxacin, ciprofloxacin and florphenicol. In contrast, they were highly resistant for ampicillin, methicillin and penicillin-G followed by amoxicillin, oxacillin and cefotaxime.
Enterococcus species are considered as a major etiological agent of nosocomial infections, they a... more Enterococcus species are considered as a major etiological agent of nosocomial infections, they are commonly isolated from different sources of food. So, this study was conducted for detection of Enterococcus spp. from fish (n=10), herbs (n=30), drinking water (n=20), dairy products (n=30), and meat products (n=20) samples from Cairo Governorate, Egypt. PCR was done for detection of some antibiotic resistant genes (mphC, norA, tetK, floR, vanA). The results revealed that from 110 food samples analyzed, 11.8% were positive for Enterococci. Moreover, E. faecalis was detected by percentage 53.8%. E. faecalis isolates were resistant to vancomycin (100%), erythromycin (57.1%), ciprofloxacin (14.3%), chloramphenicol (42.9%), tetracycline (57.1%), while none of the E. faecalis isolates were resistant to ampicillin, penicillin or norfloxacin. Genotypic characterization revealed that tetK and floR genes were present in the all 7 E. faecalis isolates. While, vanA gene was detected in 3 isolates, and mphC gene was detected only in 2 isolates. The results of our investigation indicated high levels of contamination with multi-resistant E. faecalis strains of serious concern with the isolation of strains resistant to vancomycin, considering that vancomycin is the alternative agent for patients who are intolerant to penicillin or who have Enterococci infections with high level resistance to penicillin.
The present study was conducted on 250 random samples of meat and meat products (minced meat, kof... more The present study was conducted on 250 random samples of meat and meat products (minced meat, kofta, beef burger and sausage, 50 for each) collected from small retails and different supermarkets at Kaliobia Governorate, Egypt for inspection of Lactobacillus (LAB) species and studying the bioactivities of them. The results revealed that 252 strains of LAB strains were recovered from 250 examined samples. P. acidilactici were the most isolated (102 = 40.5%) followed by L. plantarum (76=30.2%), L. acidophilus (38=15.1%); L. brevis (19=7.5%) and P. pentosaceus (17=6.7%). All of them produced bacteriocin and biosurfactant that inhibited the growth of tested pathogenic bacteria. In addition, most isolated strains had the ability for biofilm production which able to inhibit the biofilm formation of tested pathogenic strains. Moreover, the sequences obtained for 16S rRNA genes had accession numbers MK850930 and MK852180, MK852397 and MK852398, MK806485 and MK850564, MK871658 and MK871674, MK852683 and MK852691, and were 91% to 98% identical to the corresponding GenBank sequences.
Frozen fish are exposed to many risks of contamination during long chain of catching, transportat... more 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. fluorescence AB susceptibility
In this comparative study, five Brucella strains were morphologically, serologically, and biochem... more In this comparative study, five Brucella strains were morphologically, serologically, and biochemically identified using media different stains and antibiotics. Multiplex PCR was used to differentiate between the Brucella strains and Yersinia enterocolitica O: 9. five primer sets were designed to the most specific variable regions of the different Brucella species. The results of multiplex PCR were very successful and accurate in terms of characterization and typing of the Brucella vaccine, reference and wild Brucella strains from Yersinia which gave false positive while using tests based on anti-LPS antibodies but with PCR for DNA of Yersinia gave negative. The molecular typing of the Brucella strains by multiplex PCR had several advantages over the use of the conventional methods being very fast, precise, easier, more sensitive, and economic and could be applied on minimal sample preparation. The development of this PCR method is the first step toward the development of a novel kit for the molecular identification of Brucella strains from other Gram-negative bacteria.
The aim of this study was to isolate Salmonellae from 579 birds (348 chickens, 104 ducks, 30 turk... more The aim of this study was to isolate Salmonellae from 579 birds (348 chickens, 104 ducks, 30 turkeys, 50 quail, 30 pigeons and 17 geese) from 4 Egyptian Governorates. The Samples collected from internal organs (liver, cecum, spleen and heart) were examined bacteriologically and serologically. Sixty-three (10.9%) out of 579 birds were found positive while 516 (89.1%) birds were negative for Salmonella isolation. The number and percentage of positive chickens, ducks, turkeys, quails, pigeons and geese were 43 (12.4%), 10 (9.6%), 3 (10%), 5 (10%), 2 (6.7%) and 0 (0%) respectively. In this study,
A total of 150 random samples were collected from different supermarkets and retailers of differe... more A total of 150 random samples were collected from different supermarkets and retailers of different sanitation levels at Gharbeia governorate, Egypt as follows 70 samples of minced meat, 40 samples of sausage and 40 samples of beef burger. The aforementioned samples were subjected to bacteriological and serological applications to assess the prevalence of S. Enteritidis. The obtained results revealed that the incidence of S. Enteritidis in the examined samples of minced meat, sausage and beef burger were 1/70 (1.4%), 1/40(2.5%) and 0/40(0%) respectively. The isolates were submitted for serological analysis and revealed that Salmonella Enteritidis O 1,9,12 ad monophasic H:g, m. The antibiogram sensitivity test was applied upon the two isolates of S. Enteritidis and revealed that they are sensitive to chloramphinicol, amoxicillin, levofloxacin, ciprofloxacin, enrofloxacin and gentamycin but they were resistant to oxytetracycline. The virulence genes of S. Enteritidis isolates were determined by using multiple PCR technique for the two serologically detected Salmonella Enteritidis by using the following genes inv A, sef A, sop B and bcf C at 284bp, 310bp, 517bp, 467bp, respectively.
The present study was carried out to evaluate the antimicrobial activity of ethanol plant extract... more The present study was carried out to evaluate the antimicrobial activity of ethanol plant extract of rosemary (Rosmarinus officinalis) and / with five antimicrobial agents of different mechanisms (oxytetracycline Hcl, amoxicillin, cefquinome, sulphaquinoxaline and danofloxacin) against field strain of S. aureus. By using agar well diffusion method, the mean zone of inhibition (mm) of ethanol extract
The study was performed on 225 chicken (72 diseased and 153 freshly dead ones) from different 45 ... more The study was performed on 225 chicken (72 diseased and 153 freshly dead ones) from different 45 commercial broiler farms (1-30 days old) at Dakahlia Governorate for mycotic infection. Samples were taken from these chickens (lung; air sac; crop; liver and brain from each chicken) after clinical and postmortem examination for mycological examination. The results revealed that, fungi were isolated from 503 positive samples (44.7%); represented as 119 positive samples (10.6%) from diseased chickens and 384 positive samples (34.1 %) from freshly dead ones.651 fungal isolates (moulds and yeasts) were isolated from 1125 samples, where 219(33.6%) were isolated from lung samples followed by 199(30.6%); 119(18.3%); 91(14.0%) and 23(3.5%) from air sac, liver, crop and brain samples respectively. Moreover, A. fumigatus was the most isolated one 141(21.7%) followed by A. flavus 126(19.4%); A. niger 111(17.1%); Rhizopus spp. 67(10.3%); C. albicans 58(8.9%); Mucor spp. 42(6.5%); Penicillium spp. 32(4.9%); A. terreus 20(3.1%); Fusarium spp.19 (2.9%); C. krusei 13(2.0%); C. tropicalis 9(1.4%); A. candidus 9(1.4%) and finally A. ochraceus 4(0.6%). PCR using fungus-specific universal primer pairs (ITS1 and ITS4) was used for identification and genotypic characters of Aspergillus species (A. fumigatus; A. flavus and A. niger).They were able to successfully amplify the ITS1-5.8S rDNA region of all tested Aspergillus isolates, providing a single PCR product of about 600 bp for all tested isolates PCR using fungus-specific universal primer pairs (ITS and RPS) was used for identification and genotypic characters of C. providing a single PCR product of about 109 bp .Sequence of the internal transcribed spacer 1 reigon (ITS-1) of isolated A. flavus Gene Bank accession number for studied nucleotide sequence (Bankit 1867303) is KM 983253. The sequences obtained for ITS-1 region were more than 98% identical to the corresponding GenBank sequences (accession no. DQ467968.1; KC994648.1; LN482516.1 and KP689246.1). Sequence of the internal transcribed spacer 1 region (ITS-1) of isolated C. albicans. Gene Bank accession number for studied nucleotide sequence (Bankit 1867955) is KM 983254. The sequences obtained for ITS-1 region obtained were more than 95% identical to the corresponding GenBank (accession no. M87288.1; gb|S71769.1|S71769; emb (AL033396.1) and AP006852.1).
A total of (236 swabs) fecal samples from El-Menofiya and El-Kalubia Governorates, as sporadic ca... more A total of (236 swabs) fecal samples from El-Menofiya and El-Kalubia Governorates, as sporadic cases of were subjected to bacteriological, biochemical, serotyping, sensitivity testing and PCR detection of resistance genes for β-Lactames and Quinolones. Ten isolates of Salmonella species were identified, as 7 isolates (4.7%) from calves (Salmonella Typhimurium, Salmonella Enteritidis, Salmonella saintpaul, Salmonella Langeveld and Salmonella Havana), 2 isolates (3.6%) from lambs (Salmonella Typhimurium, Salmonella Bardo) and one isolate (3.3%) from goat-kids (Salmonella Enteritidis) Salmonella isolates sensitivity to β-lacatmes was 0.00% for ampicillin, penicillin G, piperacillin, cephalexin, cefoxitin, ampicillin-sulbactam, ceftazidime, 10% for ceftriaxone and 20% for amoxicillinclavulanic acid, increased to 60% and 100% for aztreonam, imipenem respectively. Susceptibility of isolates to quinolones were 10% for ciprofluxacin, 20% to nalidixic acid, and 100% for each of norfloxacin and levofloxacin. PCR study showed beta-lactamase encoding gene, bla (TEM-1), was identified in 90% and the extended-spectrum beta-lactamase, (CIT) in 20% while bla (SHV) and acc in 80% of isolates. The detection was 50%, 40%, and 0.00% for FOX, MOX, and bla (OXA-1) genes, respectively. Plasmid-mediated quinolone resistance, qnrB, qnrS were detected in 80% while aac(6')-Ib-cr, was only in 50% of the isolates. Results showed a high incidence of β-lacatmase than quinolones resistance genes and higher isolate susceptibility to quinolone than to β-lacatmes which indicates higher efficacy and validity of quinolones. Results indicates quinolones LEV and NOR in addition to βlacatmes imipenem are the drugs of choice for suspected salmonella cases.
Ten isolates of Salmonella species were obtained from sporadic cases of profuse diarrhea in new b... more Ten isolates of Salmonella species were obtained from sporadic cases of profuse diarrhea in new born ruminants from El-Menofiya and El-Kalubia Governorates in Egypt. These isolates were as follow 7 isolates from calves (Salmonella Typhimurium, Salmonella Enteritidis,two were Salmonella Saintpaul, two were Salmonella Langeveld and Salmonella Havana) ,two isolates from lambs (Salmonella Typhimurium, Salmonella Bardo) and one isolate from goat-kids(Salmonella Enteritidis). Salmonella isolates resistance to aminoglycosoides was 0.00% for Amikacin, 30% for tobramycin and 50% for streptomycin. Susceptibility of isolates to tetracycline was 50% for doxycycline. Results showed a high incidence of aminoglycosoides resistance gene aadB in 100% of the isolates while aadA2 genes in 40% of the isolates. Incidence of tetracycline resistance genes was 0.00% for tetA(B) and 80% for tetA(A). The difference between the results of this study and those from other regions in Egypt necessitate a complete survey overall the country to make a complete and clear map of salmonella servoars, their antibiotics susceptibility and in molecular characterization of resistance determinants in each region.
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