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Isolation and Molecular Identification of Avibacterium paragallinarum in Suspected Cases of Poultry
2022
Heba Badr | Heba Roshdy | Walid Kilany | Wael Elfeil | Ahmed Sedik | Wafaa Mohammed | Azhar Shalaby
Infectious Coryza (IC) is an infectious upper respiratory disease of chickens and birds, caused by Avibacterium paragallinarum (Av. Paragallinarum). It is characterized by facial swelling and nasal discharge. Here, the study aimed to identify the causative agent of infectious coryza from diseased cases using conventional and molecular methods, forty-one different samples (swollen heads and nasal discharges) from layers and broiler chickens were examined bacteriologically for isolation of Av. Paragallinarum. The total isolation percentage was 9.75%, and the percentage was 66.7% from total layer samples. The samples underwent examination by polymerase chain reaction (PCR) using HPG-2. Three samples were confirmed by PCR with a percentage of 7.3% (3/41). Multiplex PCR was used for typing the tested strains using HMTp210 gene thus, all strains were of type B. sequence analysis showed a maximum identity percentage (100%) between the tested strains of Av. paragallinarum. In conclusion, typical signs for IC diseases were appeared and identified bacteriologically as Av. paragallinarum also three strains were only confirmed by PCR and identified type B strains. PCR technique is a reliable and sensitive test for confirmation.
Show more [+] Less [-]Genetic Assessment of Shiga Toxin and Antibiotic Resistance of E. coli Isolated from Milk of Cows infected with Sub-clinical Mastitis
2022
Khaled A.S. El-Khabaz | Lamia M.T. Elshrief | Enas Elmeligy
Bovine subclinical mastitis was one of the most important health problems facing dairy industry, its impact exceeded the economic aspects and extended to potential negative effects on human health. The current study aimed to determine the prevalence of E. coli as an important mastitic pathogen and identify some of its most important virulence gene as well as their antimicrobial resistance profile. In the present study E. coli was isolated and biochemically identified whereas out of 100 subclinically mastitic milk samples was nine samples were positive for E. coli with 9% prevalence rate. Serotyping of these isolates declared that 3 isolates were serotype O26:H11, 2 isolates in serotype O91:H21 and 1 isolate in each of serotypes O55:H7, O128:H2, O146:H21 and O124. Antimicrobial resistance profile of the obtained isolates showing that all the isolates were 100% resistant to both erythromycin and streptomycin, while 88.9% (8/9) were sensitive to gentamicin. The presence of 3 important virulence factors including shiga toxin1(stx1), shiga toxin 2 (stx2) and intimin (eae) genes, among the obtained isolates was reported using PCR. Molecular investigation revealing that 2 isolates contain all studied virulence genes (stx1, stx2 and eae), 3 isolates contain (stx1 and stx2), while stx1 was detected solely in 2 isolates, also 1 isolate contain only stx2 and lastly 1 isolate was negative for any of the studied virulence factors. In a conclusion, there was a 9% prevalence rate of E. coli in subclinically mastitic milk samples in the current study, indicating its importance as a mastitic pathogen. The shiga toxin genes (stx1 & stx2) are widely distributed among E. coli isolates, while the intimin (eae) gene is less prevalent in comparison to shiga toxin genes. Also the recorded high multidrug resistance rate among the isolates posing threat to human health though entrance of these strains into the human being food chain whereas the isolated E. coli strains had the highest resistance to erythromycin and Streptomycin (100%), followed by Clindamycin (77.8%), Nalidixic acid (66.7%), and Gentamicin (11.1%) was the lowest.
Show more [+] Less [-]The Occurrence of Low Pathogenic Avian Influenza H9 Viruses in Broiler Farms Within Ismailia Province, Egypt
2022
Mostafa M.S. Shelkamy | Hanan M.F. Abdien | Dalia M. Hamed | Mohamed A. Soltan | Mona S. Abdallah
The Egyptian broiler farms are easy prey for the low pathogenic avian influenza (LPAI) H9 viruses that cause great economic losses despite their weak nature. This study focused on the occurrence and molecular characterization of the LPAI H9 viruses circulating in commercial broiler farms in different villages of Ismailia Province during 2020-2021. Tracheal, cloacal swabs, and tissue pools were collected from 34 broiler farms suffering from respiratory signs. In this study, LPAI H9 was detected in 41.17% of farms (14/34) using real-time PCR. The positive samples were propagated in 11-day-old embryonated chicken eggs (ECEs) and three isolates were selected for partial hemagglutinin (HA) gene sequencing. Sequence analysis results showed an obvious genetic evolution in comparison with the original virus (A/quail /Egypt/113413v /2011) that shared 94-94.3% and 96.7-97.1% homology at the level of nucleotides and amino acids respectively. The identity percent of the isolates with five selected commercial vaccine seed viruses revealed a higher identity with the local vaccines than the imported ones. The molecular analysis revealed 14-15 mutations in the amino acid residues with genetic stability in the main sites in comparison with the reference Egyptian strain. Phylogenetically, the H9 isolates were grouped in the G1 lineage similar to the Middle East circulating viruses with close phylogeny to Israeli viruses. The continuous genetic evolution of H9 viruses detected in this study necessitates regular virus monitoring for better control.
Show more [+] Less [-]Prevalence of Antibiotic-Resistant Vibrio Isolated from Some Marketed Fish in Egypt with a Decontamination Trial by Lemon juice
2022
Alaa Eldin M.A. Morshdy | Ahmed R. El-Ghandour | Mohamed A. Hussein | Rasha M. El Bayomi
Vibrio species are major sea foodborne pathogens that cause gastroenteritis as a serious disease of human public health due to the consumption of undercooked or raw fish. In the current study, a total of 100 fish samples (Nile tilapia, Nile perch, Meagre, and Sea bass) were collected randomly from retail markets in Egypt to investigate the prevalence of Vibrio species. The results revealed that Vibrio species isolated with an overall percentage of 52% of all examined fish. Bacteriological and chemical examinations revealed 42.3% V. parahaemolyticus, 26.92% V. mimicus, 19.23% V. alginolyticus, 9.62% V. vulnificus and 1.92%V. cholera. Antibiotic sensitivity declared high resistance of the isolates to different antimicrobial agents used in Egypt including Ampicillin (100%), Nalidixic acid (88.3%), Streptomycin (84.2%), Sulphamethoxazol (70.7%) and Oxytetracycline (64.8%) and it had sensitivity to Amikacin (94%), Ciprofloxacin (70.5%), Gentamicin (58.9%) with an average MAR index of 0.576. By polymerase chain reaction, all examined Vibrio isolates were positive for 16SrRNA specific for Vibrio spp. and harbored toxR gene virulence gene. Finally, dipping of tilapia in lemon juice 5% for 2 h reduced V. parahaemolyticus count by 0.42log cfu/g (62.08%). Consequently, hygienic measures should be approved to control the contamination of fish in the markets and the aquatic environment. Regular monitoring of fish for antibiotic resistance by Vibrio species, and their molecular characterization is necessary to improve the safety of seafood. Dipping fish in lemon juice is an efficient strategy for reducing V. parahaemolyticus load in fish.
Show more [+] Less [-]Antioxidant and Histopathological Effect of Galantamine Against Paracetamol-Induced Side Effect
2022
Gamal Shams | Shahd El-Mesiry
The goal of this research was to find out more about the possible protective role of galantamine (GAL) (0.3 mg/kg P.O) for 28 successive days against paracetamol (PCM) toxicity that was administrated on day 29 of the experiment, at day 30, blood sample were collected for evaluation of antioxidant enzymes and tissues for histopathological studies. Oxidative stress biomarkers which included serum superoxide dismutase, glutathione peroxidase, catalase, and malondialdehyde and histopathological studies indicated that GAL has protective effects by prohibiting the improvement in oxidative stress biomarkers and improved the histopathological lesions seen in livers and kidneys in vivo model against hepatic and renal toxicity induced by PCM in rats.
Show more [+] Less [-]Effect of Food Safety Management Practices on Milk Quality and Subclinical Mastitis in Dairy Cow Farms
2022
Eman Maher Tawfik | Mohammed Elsherbini | Nahed Gomaa Kasem | Marwa Fawzy ElMetwaly Ahmed | Adel Abdelkhalek
The study aimed to investigate the effect of management practices based on the principles of hazard analysis critical control points system application in dairy farms on bulk milk tank quality and the subclinical mastitis prevalence. The study was conducted on two dairy farms located in Dakahlia Governorate, Egypt using observation and questionnaire. Furthermore, cow hygiene scoring, subclinical mastitis prevalence using California Mastitis Testing, and electrical conductivity were evaluated. In addition, the organoleptic, chemical, and microbiological quality of bulk milk tanks were assessed. The results showed that farm I had better adoption of farm management practices (66.19%) than farm II (33.80%). The mean of udder and leg hygiene scores for cows showed no significant variation between both farms. The prevalence of subclinical mastitis in farm I was 0% (0/108), while it reached 6.25% (6/96) in farm II. No evidence of any abnormality during organoleptic examination on both farms. Referring to the chemical analyses, there was a higher significant difference between protein and SNF (p<0.05) in farm I than in farm II. However, this was not the case for fat, in which farm II showed a higher significance (p<0.05). Furthermore, farm I showed a significantly lower (p<0.05) somatic cell count. On the other hand, the total bacterial count (TBC), titratable acidity, and pH had no significant difference in both farms. Finally, these ensure the importance of hygiene management practices for udder health and milk quality.
Show more [+] Less [-]Microbial Profile of Imported Carcass under Chilled Storage
2022
Esraa F. Hussein | Ali Meawad Ahmed | Hanan A. Elghayaty | Heba Mohamed Shaheen
Bacterial contamination has been proven to be common in a variety of foods, especially meats. For this reason, this study was conducted to evaluate the bacteriological quality of imported chilled meat traded in Port-Said markets where 64 random samples of chilled meats represented by 28 imported chilled beef meat samples from lots arrived at Port-Said port (un-marketed) and 36 imported chilled beef meat samples collected from retailed markets at Port-Said governorate (marketed). Samples were analyzed for their total aerobic count., Enterobacteriaceae count, E. coli, total staphylococcus, and S. aureus counts and detection of salmonellae. The total bacterial count recorded an average of 10.73x104 and 2.5x106 in un-marketed and marketed chilled meat respectively. The results showed that 18 out of 64 meat samples were positive for Enterobacteriaceae and 6 samples out of them were unaccepted for human consumption. The incidence of E. coli was in 6 samples from the examined chilled samples, and the 6 were unaccepted. For staphylococcus, there were 24 positive samples, and 13 out of them were unaccepted and for S. aureus, 4 samples out of 64 samples were positive and 4 samples were unaccepted for consumption. Two samples out of 64 were positive for salmonella and considered unfit for human consumption. The obtained results confirmed the poor bacteriological quality of some imported chilled meat that is marketed in Port-Said retailed markets which is related to unhygienic transportation methods until reach the retailed markets.
Show more [+] Less [-]Molecular Studies on Some Emerging Pathogens in Dairy Products Retailed in Dakahlia Governorate, Egypt
2022
Dalia I. Makkia | Adel H. Mahmoud | Ali A. Bahout | Mohamed A. Bayoumi | Mohamed E. Alnakip
In this study, one hundred and ten samples (25 raw buffalo milk, 25 kareish cheese, 25 small-scale yoghurt, and 35 small-scale ice cream) were collected randomly from local markets, dairy shops, and supermarkets from different localities in Dakahlia Governorate, Egypt during October 2019 to March 2020. The prevalence rates of foodborne pathogens including E. coli, Salmonella spp., and Aeromonas hydrophila (A. hydrophila) were examined. Molecular confirmation and detection of toxin-producing and drug-resistance-related genes were carried out using PCR. E. coli was isolated from the examined raw milk, ice cream, Kareish cheese, and yoghurt at 72%, 25.71%, 40%, and 44%, respectively, while Salmonella spp. was isolated at 36%, 4%, and 24% from the examined raw milk, Kareish cheese, and yoghurt samples, respectively. Aeromonas hydrophila was isolated at 36% from the examined raw milk samples, 8% from Kareish cheese samples, and 76% from the examined yoghurt samples. Twelve out of 98 E. coli isolates, 18 out of 30 Salmonella isolates, and 24 out of 50 A. hydrophila were multidrug-resistant, respectively. The most resistant antibiotics were ceftriaxone and tetracycline. All examined E. coli, Salmonella spp., and A. hydrophila isolates contained blaTEM and TetA(A) resistance genes. 66.7% of the examined E. coli isolates harbored stx1, while stx2 was absent in all examined E. coli isolates. All examined Salmonella spp. isolates contained both stn and avrA virulence genes. All examined A. hydrophila contained the Aerolysin, but just 40% contained the hemolysin virulence genes. Therefore, it is necessary to reduce the excessive usage of antibiotics in dairy farms and to apply strict hygienic measures to inhibit microbial contamination of dairy products intended for human consumption.
Show more [+] Less [-]Control of Aflatoxin Residues in Broiler Chicken Using Saccharomyces cerevisiae Fortified Ration
2022
Fadwa Fathy Mahmoud | Eman Abdelnaby Ahmed | Ali Meawad Ahmed | Nada Ibrahim Hussein Ahmed
The current research was designed to examine the protective effect of probiotic-fortified ration against aflatoxin B1 (AFB1) toxicity and its residual level in broilers' edible tissues. Sensitive high-performance liquid chromatography coupled with a fluorescence detector (HPLC-FLD) was used for measuring the toxin. Ninety, one-day-old Cobb chicks were allocated into three equal groups (n=30) with three replicates per group. The first control group (G1) was fed a balanced basal diet only and the second group (G2) received AFB1 (2 mg/kg basal diet), while the third group (G3) received a combination of AFB1 (2 mg/kg basal diet) with Saccharomyces cerevisiae (SC; 1.5 g/Kg basal diet). Experimental birds were monitored for 6 weeks, their growth performance was then compared. AFB1 residue was assessed in the meat and liver sample. AFB1 resulted in a significant (P<0.05) reduction of growth performance parameters such as body weight and carcass yield in comparison to the control and SC supplemented groups. Moreover, AFB1 residue significantly (P<0.05) diminished in SC fortified group when compared with the AFB1 group. In conclusion, probiotics such as Saccharomyces cerevisiae could be considered as a potential feed additive and a growth promoter. Besides, its role in controlling AFB1 residue in the edible tissues of boiler chicken.
Show more [+] Less [-]Virulence Genes of Multi-drug Resistance Pseudomonas species Isolated from Milk and Some Dairy Products
2022
Heba Gamal | Mohamed El-Diasty | Amany Dapgh | Mohammed El-Sherbini | Amira El-Baz | Adel Abdelkhalek
Pseudomonas species is one of the psychotropic bacteria that can survive in low-tempered milk and dairy products besides producing heat-resistant spoilage enzymes. In this study, one hundred and fifty samples of milk and some dairy products were analyzed. The overall prevalence of Pseudomonas spp. was 44.66% (0% pasteurized milk, 16% butter, 20% pasteurized cream, 48.5% Talaga cheese, 50% bulk milk tank, 66.6% raw market milk, and 70% in raw cream). From 67 positive samples, eighty-three isolates were confirmed biochemically as Pseudomonas spp. The most prominent species were P. aeruginosa, then P. fluorescence, P. Fragi, P. psychrophile, P. proteolytica, P. alcaligens, P. lundensis, and P. brenneri by a percent of 38.5%, 37.5%, 10.8%, 6%, 2.4%, 2.4%, 1.2%, and 1.2%, respectively. Fourteen antibiotic discs were selected to measure the antimicrobial susceptibility of 59 isolates of Pseudomonas spp. The higher antimicrobial resistance was against Ampicillin (100%) followed by Colistin (98%), while the antibiotic sensitivity was higher against Imipenem (96.6%) then Meropenem (91.5%). The average MAR index of isolated Pseudomonas spp. was 0.462. Ten isolates of antimicrobial resistance serotypes of P. aeruginosa were O11: E, O8: C, O5: B, O4: F, and O2: B. Molecular identification of P. aeruginosa, P. fluorescence, and P. fragi was carried out using polymerase chain reaction (PCR) to determine their virulence genes (LasB, ExoS, pilB for P. aeruginosa, aprX for P. fluorescence and carA gene for P. fragi). High levels of antimicrobial-resistant (AMR) Pseudomonas spp. threaten public health and cause global concern. The economic and public health impacts were discussed.
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