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The present study aimed to determine some virulence-associated genes and antimicrobial multidrug resistance of Clostridium perfringens recovered from herbs and spices widely distributed in the Egyptian market. C. perfringens virulence and resistance factors were determined using PCR targeting the netB, tpeL, ermB, bla and tetK genes. Thirty three out of 392 samples (8.42%) from herbs and spices submitted to our laboratory for bacteriological screening were positive for presence C. perfringens. PCR results for the tpeL gene in isolated C. perfringens revealed 9 out of 33 (27.3 %) of isolates, while netB was not detected. The isolates were resistant to Clindamycin, Vancomycin, tetracycline, and erythromycin with inhibition zones of 6.28 ± 0.63, 8.78 ± 0.41, 9.63 ± 0.63, and 9.84 ± 0.66 mm, respectively. The genes mentioned above were selected to correspond to the ineffective antimicrobials; ermB for erythromycin, tetK for tetracycline, and bla for the remainder. PCR results for antibacterial resistant genes in isolated C. perfringens revealed their presence. From 33 isolates, bla gene was detected in 21 (63.4 %), tetK in 13 (39.4 %) and ermB in only one isolate (3.03 %). Sequencing analysis was done for the bla gene as an example for the detected genes as detected at the highest incidence (63.4%). No cross-relationship was detected upon comparing incidence data of both studied virulence genes and those of antimicrobial resistance. The present findings may explain the resistance of C. perfringens to the examined antibacterials and recommend avoiding the application of them to control the microbe. In addition, the authors recommend following strict hygienic procedures during the industry of herbs and spices to ensure their clearance from Clostridium perfringens before distributing the products as food additives into the markets.

The progressive expansion of aquaculture practices led to the development of bacterial disease outbreaks, otherwise the continuous and extensive use of antibiotics to overcome these diseases. Vibrio and Aeromonas species are responsible for the most severe losses affecting the aquaculture industry in Egypt. The objective of our study to investigate the antibiotic susceptibility of Vibrio parahemolyticus and Aeromonas hydrophila species isolated from Nile tilapia and Mugil fish farms in Kafr El-Sheikh province, Egypt, and molecular detection of antibiotic resistance genes. A total of 305 samples were gathered from apparently path-genomic lesions in different organs from 100 diseased fishes, 65 isolates of Vibrio species were isolated and V. parahemolyticus was isolated with an incidence of 55.4%. Out of 100 examined fish samples 72 Aeromonas species were isolated, A. hydrophila was isolated with an incidence of 99.3 %. Vibrio parahemolyticus showed high resistance for amoxicillin and colistin followed by cefotaxime and streptomycin. Meanwhile, Aeromonas hydrophila were highly resistant to amoxicillin and tetracycline followed by streptomycin, cefotaxime, and colistin. Five isolates of V. Parahemolyticus and A. hydrophila were screened using PCR for detection of 4 antibiotic resistance genes β-lactamase resistance gene (blaTEM); aminoglycosides (aadA1); tetracycline-resistant A tetA (A) and polymyxin resistance (mcr1) which were distinguished in all five V. parahaemolyticus and A. hydrophila isolates. The high detection of V. parahaemolyticus and A. hydrophila antibiotic resistance genes in our study could pose a potential risk to public health so, control methods should be taken to prevent the emergence of antibiotic resistance.

Necrotic enteritis in chicken is caused by the Clostridium perfringens, which have been found on almost chickensand for decades antimicrobial therapy was the only strategy to control it. However the misuse of antimicrobials linked to many impacts like reduced susceptibility of C. perfringens strains and cause cross-resistance to antimicrobial agents. Thisworkdemonstrates that, the uses of herbal extracts with antibacterial effectsconsider as an effective way to prevent and reducethe Necrotic enteritis.The present study determines antimicrobial activities of methanolic, ethanolic and watery extracts of thymus vulguarisagainst Clostridium Perfringenstype Ain chicken.The antibacterial activity was assessed through measuring sensitivity of isolated strains to different extracts by agar diffusiontechniqueand minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC)by using the broth macrodilution method. Thymus vulgaris different extracts consider as effective anticlostridial herbal agent. The result of agar well diffusion assayand broth macrodilution methodshowed that, the tested strains were more sensitive to theethanolic and methanolic extracts which give the lowest (MIC) and (MBC) (32, 16 respectively) than the watery extracts (512, 256 respectively) .

Mycoplasma species are the main important causative agents causing pneumonia in cattle, there are about 200 species or more. The current work aimed to investigate various mycoplasma spp. isolated from the respiratory tract of cattle by microbial culture, conventional polymerase chain reaction technique, and target gene sequencing with phylogenetic analysis. An initial screening was done to confirm the presence of mycoplasma spp. by culture on PPLO’s agar, digitonin sensitivity and biochemical tests. 129 isolates were characterized by fried egg with depressed center colonies, digitonin sensitive and negative to glucose fermentation and arginine utilization test. Out of 305 samples, 20 samples were selected for amplification by PCR technique using Mycoplasma 16S rRNA primer. Seven samples were positive to Mycoplasma species and gave amplified band at 1013 bp. subsequently, the seven isolates were sequencing. Four sequenced isolates (EGS1, EGL6, EGL1 and EGS2) were closely related to each other and very close to Mycoplasma bovis strains and far from M. leachii 99/014/6. Two sequenced isolates (EGL2 and EGL5) were closely related to each other and very close to Mycoplasma bovirhinis strains and far from M. leachii 99/014/6. One sequenced isolate (EGL3) was very close to Mycoplasma arginini strain and far from M. bovis 99/014/6. From these results, we conclude that conventional culture methods for diagnosis either by isolation and identification of Mycoplasma is a time-consuming method to diagnose mycoplasma infection So, these methods can be replaced by PCR and genome analysis technology.

Flavobacterium columnare (F.columnare) is an important pathogen affecting gills and skin of freshwater fishes causing columnaris disease. The objective of the study to isolate F.columnare from Nile tilapia and study the prevalence of antibiotics resistance strains with genotypic identification of β-lactamase resistance genes (blaTEM, bla SHV) and tetracycline resistance gene (tetA) among isolated strains of F.columnare. Two hundred diseased fish were randomly collected from different localities throughout year (2018-2019). F. columnare showed seasonal prevalence 0.00%, 10%, 40.00%, and 20.00% in winter, spring, summer and autumn respectively with total prevalence 35/200 (17.5%). F.columnare was isolated in high percent from skin and gills and rare in internal organs. The invitro antibacterial sensitivity test was applied on 20 F.columnare isolated strains and revealed resistant to pencillin, cephalosporin, aminoglycoside, nitrofurans, polymixinB and tetracycline. The isolated strains have three antibacterial resistance genes, bla TEM with amplicon size 516bp, bla SHV with amplicon size 392bp and tetA with amplicon size 576bp. Conclusion: According to this study tested strains were resistant to multiple antibiotics and their genes were widely distributed, so phenotypic and genotypic characterization of antibiotic resistant strains of isolated F. columnare are similar.

Candidiasis is an occasional opportunistic yeast disease of importance in poultry and it has also been reported to be a disease or an intestinal infection. The present study was directed mainly to throw lights firstly on the isolation of C. albicans species of chicken and secondly to the identification of the mentioned isolates by phenotypic and genotypic methods. A total 100 of swab samples collected from a crop of chicken from different localities in Cairo and Giza governorate. The samples were investigated for yeast species contamination and mycological examination, which the isolation of four yeast genera. The most frequent yeast species isolated from such samples followed by Candida spp., Saccharomyces spp., Torulopsis spp. and Rhodotorula spp. The observe incidence of Candida species isolated from all examined samples it seems that Candida species were (n=36/60, 60%) from chicken crop swab.The incidence of Candida albicans isolated from examining samples it seems (n=8/36, 22.2%) from crop swab. Early identification of yeast to the genus and species level is necessary for effective antifungal therapy, and can also facilitate control of infections. Explained that Cross-contamination is a particular problem and several recommendations have been published to control pathogens.

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.

Frequent use of antimicrobial agents may result in bacterial generation resistant to multiple antibiotics that affect on public health hazard. Our Study aimed to isolation of Edwardesiella species from fresh Oreochromis niloticus and Claris gariepinus(50 of each ) which were collected from fish farms and markets in Kafrelsheikh governorate. Atotal of 450 tissue samples involving liver, kidney, spleen, intestine from both fish species and gills from O. niloticus. Isolation and identification of the bacterial pathogens by traditional methods then antibiotic sensitivity test and resistance gene were detected. 23 isolates were obtained and distribution of Edwardesiella spp. among examined organs indicated that 9/100 (9%) isolates were detected from spleen, 6/100 (6%) from liver, 5/100 (5%) from intestine, 2/100 (2%) from kidney and 1/50 (2%) from gills. The predominant species were E. tarda so, 8 isolates were tested for 13antibiotic agents. The resistance were recorded in all 8 strains for Amoxicillin and Flumox, in 7 from 8 strains for Ampicillin, 6 from 8 for Cefotaxime, in 2 from 8 for Oxytetracycline and Streptomycin . Resistance profile genotypically in 3 isolates for β-lactamases( blaTEM, blaCTX) aminoglycosides (aada1)and tetracyclines (tetA(A))genes were 100% in all isolates. In conclusion presence of antibacterial resistance indicates misuse of antibacterial agents and affect on public health.

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 kobiba–shami 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.

The present study was performed on 250 random samples of fresh meat and meat products viz: Beef burger, kofta; minced meat and sausage (50 for each), were collected from different shops (25gm from each sample) at Kaliobia Governorate, Egypt, for detection 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, isolation of 90 (36.0%) isolates of S. aureus isolates (41/16.4%); E. coli (25 /10.0%); B. cereus (21/8.4%) and Salmonella (3/1.2%) were recovered from 250 samples. ,Seven serogroups (O55:H7; O111:H4; O125:H18; O126:H7; O128:H27; O142:H2and O158:H2) were detected in 25 isolated E. coli strains. The antibiotic sensitivity tests for the isolated strains show multiple antibiotic resistances (ampicillin; methicillin; oxytetracycline; amoxicillin; streptomycin; erythromycin; doxycycline and cefotaxime). Therefore, E. coli; S. aureus and B. cereus strains especially antibiotic resistances ones are meat-borne pathogens of public health important.

Campylobacteriosis incidence and proportion of Campylobacter strains resistant to antibiotic has been increasing 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 is isolation and biochemical characterization of Campylobacter from poultry. Campylobaceter isolation have been done on MCCDA media, motility have been detected by phase contrast microscope , morphology have been detected by Gram stain, and confirmed by MALDI-TOF MS and molecular confirmation by using cPCR by amplification of 16SrRNA gene. A total of 102 isolate of Campylobacter were isolated from (farm and small backyards located in different sources in El-Kalyobia , El-Monofia and El-sharkia Governorate) by conventional bacteriological methods from which 40 were C.jejuni and 62 were C.coli. All isolats have been confirmed as Campylobacter by MALDI-TOF MS and detection of 16SrRNA were detected by cPCR in all isolates. PCR is a useful molecular tool for identification and confirmation of Campylobacter, its Rapid, sensitive and specific than the culture methods the only disadvantage is that is expensive.

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, Egyptfor inspection of Lactobacillusspecies 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 produce 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 rRNAgenes 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 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 Saurus and Mackerel (50 of each) were collected from different fish markets in El- Menofia governorate. They were subjected to bacteriological examination for isolation of Psychrotrophic and Pseudomonas species. The bacteriological examination revealed the mean values of total Psychrotrophic count in the examined samples of Saurus was 1.05×109±1.4×108 and 1.013×109±1.6×108in Mackerel. The incidence of Pseudomonas species was 40%, 50% of the examined samples of Saurus and Mackerel, respectively. The incidence of identified Pseudomonas spp isolated from the examined samples of frozen fish were Ps. aeruginosa, Ps. dimenuta, Ps. fluorescence, Ps. putida and Ps. fragi. The Pseudomonas spp were resistant to Chloramphenicol and Nalidixic acid. In contrast they were sensitive to Gentamycin except Ps. fluorescence .

The present study was conducted on 250 diseased Nile tilapia (Oreochromis niloticus) fish of various sizes were 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 pathognomic 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 samples then 35 kidney samples ; 32 intestine samples 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.

Bovine mastitis caused by Mycoplasma bovis represents a major problem for dairy industry all over the world. Although Mycoplasmas were identified in Egypt decades ago, the prevalence of Mycoplasma bovis mastitis is not frequently investigated. The current study was designed to monitor the prevalence of Mycoplasma bovis in clinical bovine mastitis milk in Egypt. Clinical mastitis milk samples (n=703) were collected between 2016 to 2019 from different dairy farms located in different governorates in Egypt including, Giza (227), Alexandria (357), Dakahlia (78), Buhayrah (27), Ismailia (14) and presented for Mycoplasma isolation using conventional cultural method followed by molecular identification of Mycoplasma bovis using PCR targeting mb-mp 81 gene of M. bovis. From the examined samples (n=703), Sixty-three (8.96%) were positive for Mycoplasma isolation. Among the 63 Mycoplasma isolates, 53 were identified by PCR as Mycoplasma bovis representing 84.12% of the recovered Mycoplasma isolates (n=63) and 7.53% from the total examined mastitis milk samples (n=703). From the 53 Mycoplasma bovis isolates, 17/227 (7.48%) were obtained from Giza, 13/357 (3.64%) from Alexandria, 18/78 (23.07%) from Dakahlia, 1/27 (3.70%) from Al-Buhayrah and 4/14 (28.57%) from Ismailia.

In the current study, a total of 50 sheep, 23 goat and 60 human blood serum samples were collected for serological tests. A total of 30 (22 ewes and 8 does) milk samples, 43(25 ewes and 18 does) vaginal swabs, 14 preputial swabs from rams and 60 human citrated plasma were collected for bacteriological and molecular studies. All examined sheep and goat were non-vaccinated. Ewes and does had history of abortion, while rams had no clinical signs. Samples were collected from different localities at Minia governorate. Serological results using buffered acidified plate antigen test (BAPAT), rose Bengal test (RBT), rivanol test (RT), tube agglutination test (TAT), and indirect enzyme linked immunosorbent assay (iELISA) gave 32(64%), 30(60%), 26(52%), 31 (62%), and 28 (56%) in sheep respectively, where in goat revealed 23(100%), 23(100%), 22(95.7%), 23(100%), and 22(95.7%) respectively. While human serum samples were 33(55%), 29(48.3%), 33(55%), 27(45%), and 23(38.3%) respectively. The bacteriological results revealed 35 isolates [10(45.5%), 4(50%), 4(16%), 10(55.6%), 2(14.3%), and 5(8.3%)] from ewes' milk samples, does' milk samples, ewes' vaginal swabs, does' vaginal swabs, rams' preputial swabs and human citrated plasma respectively. All 35 isolates were molecularly identified using polymerase chain reaction (PCR) as B. melitensis biovar3. Conclusively, brucellosis was endemic at Minia governorate with a high level especially in goat. Rams act as chronic carriers leading to public health hazards. B. melitensis is the prevalent serotype in Egypt. More effective preventive and control measures should be applied

Brucellosis is a major infectious zoonotic disease affecting animal and human.In animal, the disease is causing great economic losses such as abortion, still birth as well as infertility in both male and female.A cross sectional study carried out from November 2016 to May 2017 to estimate the seroprevalence of brucellosis in sheep and goatsin Benha.A total of 38 animals (20 sheep, 18 goats)were investigated. 19 serum samples (8 from sheep and 11 from goats) subjected for serological examination. The results showed that,5/8(62.5%) and 9/11(81.8%) from sheep and goats, respectively, were positive by Rose Bengal Plat test(RBPT), Buffered Acidified Plat Antigen test(BABA)and Rivanol test (RIV test).Atotal of 34 lymph nodes from seropositive sheep and goats were examined bacteriologically. 3/20 (15%) and 4/14 (28.57%) were culture positive for Brucellarespectively. None of vaginal swabs revealed culture positive results.All isolates were confirmed genetically as Brucella spp. by PCR and all strains identified asB. melitensis by AMOS- PCR.

A total of 200 random samples of fresh dairy milk (80), soft cheese (40), kariesh cheese (40) and ice cream (40) were collected from small retails and different shops at Qaliopia and Giza governorates during the period of October 2016 to January 2017 and transferred with minimum delay to laboratory for detection the presence of Listeria species. The bacteriological results revealed that, 5/200(2.5%) were Listeria monocytogenes (L. monocytogenes) includes 3/80 (3.75 %) from raw milk, 1/40 (2.5%) from each kariesh cheese and ice cream samples and 0/40(0%) from soft cheese. The results of Microgen TM Listeria-ID System revealed that all isolates were L. monocytogenes (99.92%). The PCR results for L. monocytogenes showed that all 16S rRNA were detected in five studied strains (100.0%) i.e., all studied strains were L. monocytogenes.

The present work aimed to isolate and characterize E. coli bacteria that transmitted from fish to fishhandlers in markets and farm workers, for this 200 apparently healthy Oreochromis niloticus werecollected randomly from different markets and farms and 50 human skin swabs from sellers atmarkets and workers at farms were obtained at Kafr El-sheikh governorate, Egypt. These sampleswere cultured and biochemical characters was studied. Serological identification, antibiogramactivity and molecular characterization of some virulence factors were detected. Results, theprevalence of Escherichia coli was 17.5% in Tilapia while its prevalence in human was 24%.Escherichia coli serotypes from fish were O153 :H2, O1 :H7, O125 :H21 and O78. Only one from fourisolates of Escherichia coli was positive to eaeA gene and the four isolates were negative to hlygene.

Listeria monocytogenes has become increasingly important as a food-associated pathogen. It cancause a rare but serious disease called listeriosis with high fatality rates (20–30%) compared withother foodborne microbial pathogens. To estimate the incidence and levels of Listeria spp. indifferent food sources, A total of 200 random samples which collected from different sources 65samples isolated from poultry , 45 from poultry byproducts (Frozen chicken meat balls (kofta),Frozen chicken burger and frozen chicken sausages), 20 from pasteurized milk ,30 from milkproducts (Feta cheese ,cream cheese )and 40 from poultry eggs .Only 7 of total 200 samples (14%)that collected from suspected isolates and detected on Oxford agar, PALCAM agar and ALOA agarwere suspected to be Listeria spp. In poultry, poultry byproducts, pasteurized milk, milk byproductsand poultry eggs, listeria percentage were 0/65(0%), 4/45(1.8), 0/20(0%), 0/30(0%), 3/40(1.2%)isolates, respectively. The in-vitro antimicrobial sensitivity test showed that the isolated L.monocytogenes were sensitive to Sulphamethoxazole /trimethoprim, Gentamycin, Ceftazidime andCephazoline followed by Sulphamethoxazole, ceftriaxone , weak sensitivity to Penicillin . Whilethey were resistant to Erythromycin, Clindamycin, Ampicillin. The results of virulence tests forisolated listeria strains appeared that all of L .monocytogenes were virulent strains as all of themwere positive to CAMP test; showed narrow zone of β–hemolysis on sheep blood agar .The PCRresults for Listeria isolates showed that all strains are L. monoctogenes and have Inl A and hly Agenes positive as virulence genes while negative Inl B, iap and prf A genes.

Antimicrobial activity of methanol piper nigrum extract in combination with some antibiotics was investigated againstEscherichia coli isolated from chicken samples. The in-vitro antimicrobial sensitivity pattern of these isolates was doneand the intermediate isolates were checked for combination with methanol piper nigrum extract after detecting theirantimicrobial activities against these isolates by both disc diffusion test and MIC and the obtained data showed thatantimicrobial activities of amoxicillin, doxycycline, gentamicin, erythromycin and cefotaxime were enhanced incombination with methanol piper nigrum extract. The obtained data showed that enhancement by both disc diffusion testand MIC for methanol piper nigrum extract and each of the mentioned antibiotics in decimal assay of additivity ratio was(0.5 for amoxicillin, cefotaxime and 0.5 for extract and 0.4 for doxycycline, gentamicin, and erythromycin and 0.6 forextract) against E. coli isolates.

The contamination of milk and cheese with different types of fungi would constitute a public health hazard . These fungi may cause mycotoxicosis in human and/or spoilage of these products. This study aimed to the isolation and identification of mould and yeast in raw milk and some cheese. Sixty samples were collected from Giza Governorate, Egypt including; 20 raw milk, 20 processed cheese and 20 soft white cheese. The results reported that, overall incidence rate of the moulds in processed cheese, soft white cheese and raw milk were; 18/20 (90%), 6/20 (30%) and 16/20 (80 %), ,respectively while the mean mould counts were 1.3 × 103 ± 1.8× 10² cfu/g ,6.7 × 103 ± 2.6× 103 cfu/g and 5.9 × 103 ± 1.3× 103cfu/ml,,respectively .The isolated mould species from these samples were Aspergillus spp., Penicillium spp., Eupenicillium spp. ,Phoma spp., Geotrichum spp. and Fusarium spp. while, the yeast contamination was detected in soft white cheese and raw milk samples with incidence rate of 10/20 (50%) and 16/20 (80%); respectively, while not detected in processed cheese samples. The most predominant yeast species isolated from soft white cheese and raw milk samples were Debaryomyces hansenii, Pichia anomala, Sacharomyces cervisioe ,Candida tropicalis, Candida pseudotropicalis, Torulopsis and Rhodotorula. In conclusion, the presence of high incidence of fungi in raw milk and cheese samples would lead to a public health hazards and/or economic losses due to spoilage of these products, therefore, strict hygienic conditions are required. Keywords: Moulds, Yeasts, Raw milk , Cheese, Egypt.

Two hundred random samples of milk, kareish cheese, yoghurt and ice-cream (50 for each) were examinedmicrobiologically for the presence of Clostridium perfringens, their enterotoxigencity and their antibiotic sensitivity.Clostridium perfringens was isolated from 3 (6%) milk samples, 4 (8%) kareish cheese samples and it could not beisolated from any examined samples of yoghurt and ice-cream. The majority of C. perfringens isolates recovered frommilk and milk products were susceptible to ofloxacin, ampicillin + sulbactam and norfloxacin (100%), vancomycin,tetracycline, metronidazole and amoxicillin + clavulinic acid (83.3%) and clindamycin (66.7%). The majority wereresistant to cephalothin (100%), sulphamethoxazole + trimethoprim (83.3%), oxacillin and chloramphenicol (66.7%).Molecular studies using multiplex PCR technique for detection of alpha toxin gene and C.perfringens types "A"enterotoxin gene revealed that the 7 isolates of C. perfringens (100%) were positive for alpha toxin gene and only 2 outof 7 isolates (28.57%) were positive for enterotoxin gene

A total of 102 chicken samples were collected from different farms in Qalubia, Behera, Cairo, Assuit and MenofiaGovernorates. The samples were represented by liver, spleen, lungs, heart blood, intestine and kidneys and subjected forisolation and identification of Escherichia coli. The bacteriological examination of the samples indicated the isolation of55 E. coli represented as 31, 6, 8, 5, and 5 from Qalubia, Behera, Cairo, Assuit and Menofia, respectively. Differentserotypes of E. coli (O158, O125, O111, O27, O20, O6, O25, O26, O145, and O159) and12 untyped strains were demonstrated.Thirty-Two E. coli isolates were subjected to initial screening test for extended-spectrum beta-lactamases (ESBLs)production by disc diffusion method with various cephalosporins. The results showed that 46.9% of samples weresensitive to ceftriaxone and cefotaxime and 53.1% to ceftazidime. By double disc synergy test, 19 E. coli strains wereidentified as ESBL producers. PCR results of 32 E. coli isolates showed that blaTEM gene was detected in the genomicDNA of all isolates and in plasmid DNA of 18 isolates. While, blaSHV was detected in the genomic DNA of 12 isolatesand in plasmid DNA of 9 isolates. We can conclude from the current results that amplification of both genomic andplasmid DNA increase the positivity of detection in comparison with amplification of each of DNAs alone

Mycobacterium bovis (M. bovis) BCG belongs to Mycobacterium tuberculosis complex (MTBC), highly relatedorganisms, which are 99.9 % similar at nucleotide level and phenotypically similar. Its differentiation from other membersof MTBC by conventional methods is laborious and time consuming. PCR provides a rapid alternative method fordifferentiation between members of MTBC. It depends on identification of region of difference (RD) which have beenlost during attenuation of M. bovis. Two different BCG strains (from two sources) were confirmed as a member of M.tuberculosis (MTB) complex (MTC) and as BCG strains by PCR using primers to a region of the 16S rRNA gene that isconserved in all mycobacteria and region of difference (RD1, RD4, RD9 and RD12) respectively. DNA of Mycobacteriumtuberculosis was used as a control to compare with BCG strains. The results showed that 16S rRNA gene was present inall tested strains, while the RD1, RD4, RD9 and RD12 were absent only in BCG strains

Mycobacterium bovis BCG is attenuated strain derived from M. bovis. It has been known to prevent childhood pulmonaryand meningeal tuberculosis and to treat superficial bladder cancer as immunotherapy. Administration of BCG has beenrestricted to immune-deficient individuals since 2007 because of its combined complications. Which treated withantituberculosis drugs based on assumption is fully responsive to these drugs. BCG Connaught showed moresusceptibility to the activity of isoniazid, ethambutol, kanamycin, ofloxacin, streptomycin, amikacin and capreomycinthan BCG Pasteur growing in J774A.1 cells. While the activity of rifampicin and levofloxacin was the same between thetwo tested strains. The spectrum of intracellular drug action can be ordered based on a decreasing order of inhibitoryactivity, as following rifampin > isoniazid > ethambutol > streptomycin > kanamycin> amikacin> ofloxacin>capreomycin> levofloxacin against BCG Connaught. While rifampin > isoniazid > ethambutol > kanamycin =streptomycin > ofloxacin> amikacin> levofloxacin> capreomycin for BCG Pasteur. There is a very limited publicationsdescribe the activity of antituberculosis drugs against intracellular growing BCG strains

Aim: Participants in an unorganized gathering are potential hosts of diseases, bringing diseases from around the world to be introduced to a large at-risk population. Therefore, we investigated the gene repertoire in 29 Escherichia coli strains linked to urinary tract infection isolated from patients transferred to the hospital after attending an unorganized gathering in Cairo. Materials & methods: Virulence and resistance determinants, phenotypic antibiotic resistance, biofilm formation, their serotypes and phylogenetic relationships were analyzed. Results: The 29 tested serovars were phenotypically virulent, with the prevalence of group B2, and resistant to tetracycline, naldixic acid, ampicillin, trimethoprim, neomycin, oxytetracycline and erythromycin encoding the iss virulent gene. Conclusion: A One Health approach is a must to monitor and control E. coli urinary tract infections.

The current study was designed to examine the antibacterial effect of thyme (thymus vulgaris) on Escherichia coli in diarrheic calves and also the effect of the thyme as immunostimulant. Collection of 100 rectal swabs from 100 diarrheic calves was done for isolation and identification of E.coli and serotyping of the bacterial isolates followed by their antibiotic sensitivity testing and determination of MIC and MBC of thyme oil on them. Supplementation of thyme extract to friesian calves milk was done and examined its effect of on their immunity by antibodies titration of IgM, IgG and IgA in their serum. Results revealed that the incidence of E.coli was 58% and pathogenic E.coli was 46%.The most predominant serotype was O125. The highly resistance of the strains was against oxytetracycline (OT) and ampicillin (AMP). The thyme had antibacterial effect against E.coli strains by MIC ranged from 5 to 320 µg /ml but the MBC was negative for all strains and the thyme extract at 40 mg/kg body weight improved the immunity as it significantly (P≤0.05) increased IgM, IgG and IgA titre in calves serum.

The study was performed on 225 chicken (72 diseased and 153 freshly dead ones) from different 45commercial broiler farms (1- 30 days old) at Dakahlia Governorate for mycotic infection. Samples weretaken from these chickens (lung; air sac; crop; liver and brain from each chicken) after clinical andpostmortem examination for mycological examination. The results revealed that, fungi were isolatedfrom 503 positive samples (44.7%); represented as 119 positive samples (10.6%) from diseased chickensand 384 positive samples (34.1 %) from freshly dead ones.651 fungal isolates (moulds and yeasts) wereisolated from 1125 samples, where 219(33.6%) were isolated from lung samples followed by199(30.6%); 119(18.3%); 91(14.0%) and 23(3.5%) from air sac, liver, crop and brain samplesrespectively. Moreover, A. fumigatus was the most isolated one 141(21.7%) followed by A. flavus126(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-specificuniversal primer pairs (ITS1 and ITS4) was used for identification and genotypic characters ofAspergillus species (A. fumigatus; A. flavus and A. niger).They were able to successfully amplify theITS1–5.8S rDNA region of all tested Aspergillus isolates, providing a single PCR product of about 600bp for all tested isolates PCR using fungus-specific universal primer pairs (ITS and RPS) was used foridentification and genotypic characters of C. providing a single PCR product of about 109 bp .Sequenceof the internal transcribed spacer 1 reigon (ITS-1) of isolated A. flavus Gene Bank accession numberfor studied nucleotide sequence (Bankit 1867303) is KM 983253. The sequences obtained for ITS-1region 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 spacer1 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 than95% identical to the corresponding GenBank (accession no. M87288.1; gb|S71769.1|S71769; emb(AL033396.1) and AP006852.1).

A total of 120 random samples of meat products including luncheon, sausage, minced meat and beefburger (30 from each) were collected from different supermarkets in Gharbia Governorate. The collectedsamples were transferred directly to the laboratory to be examined bacteriologically for the detection ofB. cereus. All samples were cultured at 37°C for 24 hours aerobically on selective media (PEMBA) forisolation and purification and on sheep blood agar to observe the hemolysis properties. The resultsrevealed that the incidence of B. cereus was 20%, 36.37%, 40% and 56.67% in luncheon, beef burger,sausage and Minced meat, respectively. The suspected colonies were examined for their colonialmorphology, microscopical examination, and biochemical reactions.VITEK2 BCL Card was used formore identification of B. cereus strains. The public health hazards of the isolated strains were discussed

Background The objectives of this study were to characterize the diversity and magnitude of antimicrobial resistance among Staphylococcus species recovered from imported beef meat sold in the Egyptian market and the potential mechanisms underlying the antimicrobial resistance phenotypes including harboring of resistance genes (mecA, cfr, gyrA, gyrB, and grlA) and biofilm formation. Results The resistance gene mecA was detected in 50% of methicillin-resistant non-Staphylococcus aureus isolates (4/8). Interestingly, our results showed that: (i) resistance genes mecA, gyrA, gyrB, grlA, and cfr were absent in Staphylococcus hominis and Staphylococcus hemolyticus isolates, although S. hominis was phenotypically resistant to methicillin (MR-non-S. aureus) while S. hemolyticus was resistant to vancomycin only; (ii) S. aureus isolates did not carry the mecA gene (100%) and were phenotypically characterized as methicillin- susceptible S. aureus (MSS); and (iii) the resistance gene mecA was present in one isolate (1/3) of Staphylococcus lugdunensis that was phenotypically characterized as methicillin-susceptible non-S. aureus (MSNSA). Conclusions Our findings highlight the potential risk for consumers, in the absence of actionable risk management information systems, of imported foods and advice a strict implementation of international standards by different venues such as CODEX to avoid the increase in prevalence of coagulase positive and coagulase negative Staphylococcus isolates and their antibiotic resistance genes in imported beef meat at the Egyptian market.

Although normally regarded harmless commensals, enterococci may cause a range of different infections in humans, including urinary tract infections, sepsis, and endocarditis. The acquisition of vancomycin resistance by enterococci (VRE) has seriously affected the treatment and infection control of these organisms. VRE are frequently resistant to all antibiotics that are effective treatment for vancomycin-susceptible enterococci, which leaves clinicians treating VRE infections with limited therapeutic options. With VRE emerging as a global threat to public health, we aimed to isolate, identify enterococci species from tilapia and their resistance to van-mediated glycopeptide (vanA and vanC) as well as the presence of enterococcal surface protein (esp) using conventional and molecular methods. The cultural, biochemical (Vitek 2 system) and polymerase chain reaction results revealed eight Enterococcus isolates from the 80 fish samples (10%) to be further identified as E. faecalis (6/8, 75%) and E gallinarum (2/8, 25%). Intraperitoneal injection of healthy Nile tilapia with the eight Enterococcus isolates caused significant morbidity (70%) within 3 days and 100% mortality at 6 days post-injection with general signs of septicemia. All of the eight Enterococcus isolates were found to be resistant to tetracycline. The 6/6 E. faecalis isolates were susceptible for penicillin, nitrofurantoin, gentamicin, and streptomycin. On the other hand 5/6 were susceptible for ampicillin, vancomycin, chloramphenicol, and ciprofloxacin. The two isolates of E. gallinarum were sensitive to rifampicin and ciprofloxacin and resistant to vancomycin, chloramphenicol, and erythromycin. Molecular characterization proved that they all presented the prototypic vanC element. On the whole, one of the two vancomycin resistance gene was present in 3/8 of the enterococci isolates, while the esp virulence gene was present in 1/8 of the enterococci isolates. The results in this study emphasize the potential role that aquatic environments are correlated to proximity to anthropogenic activities in determining the antimicrobial resistance patterns of Enterococcus spp. recovered from fish in the river Nile in Giza, Elmounib, Egypt as a continuation of our larger study on the reservoirs of antibiotic resistance in the environment.

Foodborne pathogens are leading causes of illness especially in developing countries. The current study aimed to characterize virulence-associated genes and antimicrobial resistance in 30 Salmonella Typhimurium isolates of chicken and human origin at Mansoura, Egypt. The results showed that invA, avrA, mgtC, stn, and bcfC genes were identified in all the examined isolates, while 96.7% and 6.7% were positive for sopB and pef genes, respectively. The highest resistance frequencies of the isolates were to chloramphenicol and trimethoprim-sulfamethoxazole (73.3%, each), followed by streptomycin (56.7%), tetracycline and ampicillin (53.3%, each), and gentamicin (30%). However, only 2.7% of the isolates were resistant to cefotaxime and ceftriaxone each. Different resistance-associated genes, including blaTEM, aadB, aadC, aadA1, aadA2, floR, tetA(A), tetA(B), and sul1, were identified in Salmonella Typhimurium isolates with the respective frequencies of 53.3%, 6.7%, 23.3%, 46.7%, 63.3%, 73.3%, 60%, 20%, and 96.7%. None of the isolates was positive for blaSHV, blaOXA, and blaCMY genes. The results showed that the intI1 gene was detected in 24 (80%) of the examined Salmonella Typhimurium isolates. Class 1 integrons were found in 19 (79.2%) isolates that were intI1 positive. Seven integron profiles (namely: P-I to P-VII) were identified with P-V (gene cassette dfrA15, aadA2), the most prevalent profile. To the best of our knowledge, this is the first study to characterize the unusual gene cassette array dfrA12-OrfF-aadA27 from Salmonella Typhimurium isolates in Egypt.