The role of turmeric and black pepper oil nanoemulsion in attenuating cytokine storm triggered by duck hepatitis A virus type I (DHAV-I)-induced infection in ducklings.

The cytokine storm induced by duck hepatitis A virus type 1 (DHAV-1) infection significantly contributes to severe, rapid deaths and economic losses in the duck industry in Egypt. This study aimed to investigate the potential inhibitory effect of a nanoemulsion containing turmeric and black pepper oil on the immune response and pathogenesis of DHAV-1 in ducklings. A total of 105 ducklings from nonvaccinated breeders were divided into 5 experimental groups, each comprising 21 birds. The negative control group (G1) remained noninfected with DHAV-1 and nontreated with nanoemulsion, while the positive control group (G2) was infected with DHAV-1 but not treated with nanoemulsion. The other 2 groups (G3, the supplemented group which was noninfected with DHAV-1), and group 4 (the prophylactic group G4) which was infected with DHAV-1, both received nanoemulsion throughout the experiment. Group 5 (G5, the therapeutic group), on the other hand, which was infected with DHAV-1 received nanoemulsion only from the onset of clinical signs. At 5 days old, the ducklings in the positive control (G2), the prophylactic (G4), and the therapeutic group (G5) were infected with DHAV-1. All the ducklings in the infected groups exhibited depression, anorexia, and opisthotonos, and their livers displayed various degrees of ecchymotic hemorrhage, liver enlargement, and microscopic pathological lesions. Notably, the positive control group (G2) experienced the most severe and pronounced effects compared to the other infected groups treated with the nanoemulsion. Meanwhile, the viral RNA loads were lower in the liver tissues of the infected ducklings treated with the nanoemulsion (G4, and G5) compared to the positive control group G2. Additionally, the nanoemulsion effectively modulated proinflammatory cytokine expression, antioxidant enzymes, liver enzymes, and lipid profile of treated ducklings. In conclusion, the turmeric and black pepper oil nanoemulsion has the potential to be a therapeutic agent for regulating and modulating the immune response, decreasing DHAV-1-induced cytokine storms, and minimizing mortality and economic losses in the duck business. More research is needed to understand how turmeric and black pepper oil nanoemulsion alleviates DHVA-1-induced cytokine storms and lowers duckling mortality.

Detection and molecular characterization of Trichomonas gallinae recovered from domestic pigeons in Egypt.

Trichomonas gallinae is a protozoan parasite that causes canker in pigeons. Squabs (young pigeons) are frequently infected with T. gallinae and can die because of the infection, while adult pigeons can act as carriers showing no clinical signs. In the present study, 50 squabs, up to 1-month-old, were purchased from pigeon markets in different regions of the Giza governorate, Egypt. Direct wet mount preparations of the oral excretions of the squabs (mouth wash) and Giemsa staining revealed that 64% (32/50) were positive for T. gallinae. Experimental infection of ten squabs with 103 T. gallinae trophozoites/ml resulted in oral lesions on the mouth, tongue, and soft palate, with the presence of yellowish-white nodules (cheese-like) in the oral cavity on the sixth day post-infection in all squabs. A subset of five samples were cultured in modified Diamond's media, their DNA was extracted, and a portion of the ribosomal internal transcribed spacer region (ITS1/5.8S/ITS2) was amplified by polymerase chain reaction (PCR) followed by sequencing. Phylogenetic analysis of the five isolates revealed 64-91% homology with some reference isolates circulating in Egypt and related countries.

Molecular characterization of some isolates of rabbit viral hemorrhagic disease (VHD) in Egypt from 2014 to 2019.

OBJECTIVE: Rabbit viral hemorrhagic disease (VHD) is a transmittable and lethal viral illness of rabbits. In this study, genetic identification and genetic analysis of the rabbit hemorrhagic disease virus (RHDV) was made in three governorates in Egypt from 2014 to 2019. MATERIALS AND METHODS: Livers from 18 freshly dead rabbits, which was guessed to be VHD epidemics in Egypt (Giza, Menofia, and Fayoum governorates) from 2014 to 2019, were examined for RHDV. The examination was based on the hemagglutination assay (HA) test against different mammalian (human O-type and sheep) and avian (chicken and pigeon) erythrocytes, reverse transcriptase-polymerase chain reaction (RT-PCR), and sequencing of the segment of VP60. RESULTS: 33% of the examined samples' virus titers were 5 log2 to 8 log2 hemagglutination of human O-type erythrocytes when compared to 28%, 11%, and 28% of sheep, chicken, and pigeon erythrocytes, respectively. Four RHDV isolates out of eight RT-PCR positives were sequenced and phylogenetically analyzed. Sequenced isolates were designed and submitted to GenBank with accession numbers MN904506, MN904507, MN904508, and MN904509. These four RHDV isolates were related to classic G3 (GI.1d/RHDV). Twelve amino acid differences were detected between the vaccine strain sequence (Giza-2006) and RHDV isolates. Amino acid differences at 416, 423, and 476 positions seem interesting as they changed polarity that could change the protein structure and affect host interaction. CONCLUSIONS: There is antigenic variation between circulating RHVD strains and the vaccinal strain. This may be the leading cause of vaccination failure and may increase the need to check out the vaccination program against RHVD.

Vancomycin and florfenicol resistant Enterococcus faecalis and Enterococcus faecium isolated from human urine in an Egyptian urban-rural community.

Multidrug resistance is one of the top three threats to global public health. Understanding resistance of bacteria is important to help decrease resistance and improve the development of novel antimicrobial agents or other alternative tools to combat public health challenges. Thus, the goal of this study was to investigate the vancomycin and florfenicol resistance genes of five E. faecalis and 15 E. faecium isolated from patients with urinary tract infections. There were 20 Enterococcus obtained from the library collection of randomly selected private hospitals located in the city of El Qanater El Khayreya; these samples were isolated during 2017. Samples were evaluated for their phenotypic characterization of virulence factors, antimicrobial resistance and PCR was conducted to detect the prescence of the vancomycin vanABC and florfenicol resistance genes encoding the catAB, fexAB and cfu. There were six different antibiotic resistance profiles observed. The 20 isolates showed resistance to clindamycin, oxytetracycline and gentamycin. Resistance was evident to ciprofloxacin, norfloxacin and florfenicol in the absence of the cfr gene in all of the 20 Enterococcus isolates. In addition, all isolates produced biofilms and were classified as extensive drug resistant. MARindices of the isolates were >0.6. The MARindex of human isolates of enterococci suggest these pathogens originate from a high-risk source of contamination where antibiotics are often used. This information highlights a possible public health concern to the Egyptian community. The results also suggest the emergence of a linezolid sensitive-vancomycin resistant E. faecium and E. faecalis in the absence of the cfr gene.

Poultry as a vector for emerging multidrug resistant Enterococcus spp.: First report of vancomycin (van) and the chloramphenicol-florfenicol (cat-fex-cfr) resistance genes from pigeon and duck faeces.

Although commonly regarded as human and animal intestinal tract commensals, Enterococcus spp. have emerged as important nosocomial pathogens due to their intrinsic or acquired resistance to a number of antibiotics. Poultry has been suggested to be a reservoir for antibiotic resistance that may aggravate the problem of transmission of enterococci infections. Between January and December 2016, 106 Enterococcus spp. were isolated from a total of three poultry species. The collection included isolates recovered from chickens (n = 30), ducks (n = 35) and pigeons (n = 41). All enterococci isolates were screened for their ability to form biofilm. The antibiotic susceptibility was determined against 13 antibiotics using the disc diffusion method. The presence of the eight resistance genes, vanA, vanB, vanC, catA, catB, fexA, fexB and cfr was determined by PCR. All 106 isolates were resistant to clindamycin, whereas majority of isolates (>90%) were resistant to erythromycin, oxytetracycline, doxycycline, gentamycin, ciprofloxacin, norfloxacin, and vancomycin. All isolates produced biofilms and were classified as extensive drug-resistant. MARindices for all isolates was determined to be > 0.8, indicating that they have been recovered from high risk contamination sources. The cfr resistance gene was not detected in any of the 106 enterococci isolates, whereas the chloramphenicol resistance genes catA and catB were found in 18.9% (20/106) of the isolates. Interestengly, fexA 11.9% (15/106), fexB 8.7% (11/106), vanA 18.9% (20/106), vanB 25.5% (27/106), and vanC 33% (35/106) genes were also determined in our study. The present study highlights the emergence of a linezolid sensitive-vancomycin resistant enterococci, which lacks the cfr gene reporting also for the first time the detection of van, fex and cat -genes in Enterococcus species recovered from chickens, ducks and pigeons in Egypt suggesting that poultry species could be potential vectors for transmission of multidrug resistant enterococci posing a public health risk.

Prevalence of the Antibiotic Resistance Genes in Coagulase-Positive-and Negative-Staphylococcus in Chicken Meat Retailed to Consumers.

The use of antibiotics in farm management (growing crops and raising animals) has become a major area of concern. Its implications is the consequent emergence of antibiotic resistant bacteria (ARB) and accordingly their access into the human food chain with passage of antibiotic resistance genes (ARG) to the normal human intestinal microbiota and hence to other pathogenic bacteria causative human disease. Therefore, we pursued in this study to unravel the frequency and the quinolone resistance determining region, mecA and cfr genes of methicillin-susceptible Staphylococcus aureus (MSSA), methicillin-resistant S. aureus (MRSA), methicillin-resistant coagulase-negative staphylococci (MRCNS) and methicillin-susceptible coagulase-negative staphylococci (MSCNS) isolated from the retail trade of ready-to-eat raw chicken meat samples collected during 1 year and sold across the Great Cairo area. The 50 Staphylococcus isolated from retail raw chicken meat were analyzed for their antibiotic resistance phenotypic profile on 12 antibiotics (penicillin, oxacillin, methicillin, ampicillin-sulbactam, erythromycin, tetracycline, clindamycin, gentamicin, ciprofloxacin, chloramphenicol, sulfamethoxazole-trimethoprim, and vancomycin) and their endorsement of the quinolone resistance determining region, mecA and cfr genes. The isolation results revealed 50 isolates, CPS (14) and CNS (36), representing ten species (S. aureus, S. hyicus, S. epidermedius, S. lugdunensis, S. haemolyticus, S. hominus, S. schleiferi, S. cohnii, S. intermedius, and S. lentus). Twenty seven isolates were methicillin-resistant. Out of the characterized 50 staphylococcal isolates, three were MRSA but only 2/3 carried the mecA gene. The ARG that bestows resistance to quinolones, ß-lactams, macrolides, lincosamides, and streptogramin B [MLS(B)] in MRSA and MR-CNS were perceived. According to the available literature, the present investigation was a unique endeavor into the identification of the quinolone-resistance-determining-regions, the identification of MRSA and MR-CNS from retail chicken meat in Egypt. In addition, these isolates might indicate the promulgation of methicillin, oxacillin and vancomycin resistance in the community and imply food safety hazards.

Antimicrobial Resistance, Biofilm Formation and mecA Characterization of Methicillin-Susceptible S. aureus and Non-S. aureus of Beef Meat Origin in Egypt.

Methicillin-resistant Staphylococcus aureus (MRSA) have been found in various farm animal species throughout the world. Yet, methicillin-susceptible S. aureus (MSSA), methicillin-susceptible non-S. aureus (MS-NSA), and methicillin-resistant non-S. aureus (MR-NSA) were not investigated. Therefore, we persued to determine the diversity in their phenotypic virulence assay, phenotypic antimicrobial resistance profile and molecular characterization in one of the food chains in Egypt. Samples were collected during 2013 from beef meat at retail. Twenty seven isolates comprising five species (S. hyicus, S. aureus, S. schleiferi subsp. coagulans, S. intermedius, and S. lentus) were characterized for their antibiotic resistance phenotypic profile and antibiotic resistance genes (mecA, cfr, gyrA, gyrB, and grlA). Out of the 27 Staphylococcus isolates only one isolate was resistant to the 12 antibiotics representing nine classes. Raw beef meat sold across the Great Cairo zone, contains 66.7% of MRS, with highest prevalence was reported in S. aureus (66.7%), while the MRS non-S. aureus strains constituted 66.7% from which S. hyicus (60%), S. intermedius (33.3%), S. schleiferi subsp. coagulans (100%), and S. lentus (100%) were MRS. Seven S. aureus, six S. hyicus, four S. schleiferi subsp. coagulans, three S. intermedius, and one S. lentus isolates although being resistant to oxacillin yet, 11/27 (40.7%) carried the mecA gene. At the same time, the cfr gene was present in 2 of the nine S. aureus isolates, and totally undetectable in S. hyicus, S. schleiferi subsp. coagulans, S. intermedius, and S. lentus. Although, global researches largely focused into MRSA and MR-NSA in animals on pigs, the analysis of our results stipulates, that buffaloes and cattle could be MRSA dispersers and that this theme is not specific to pigs. Detection of MSSA virulence determinants is a must, as although oxacillin resistance may be absent yet, the MSSA may carry the virulence determinants which could be a source of perilous S. aureus for the human community.

Prevalence and antimicrobial resistance profile of Staphylococcus species in chicken and beef raw meat in Egypt.

Coagulase-positive (CPS) and coagulase-negative (CNS) staphylococci cause staphylococcal food poisoning. Recently, CPS and CNS have received increasing attention due to their potential role in the dissemination of antibiotic resistance markers. The present study aimed to evaluate CPS and CNS species distribution and their antibiotic resistance profile isolated from chicken and beef meat. Fifty fresh, uncooked chicken parts and 50 beef meat cuts (local n=27; imported n=23) were used. One hundred staphylococcal isolates belonging to 11 species were isolated and identified from chicken (n=50) and beef (n=50) raw meat samples. Staphylococcus hyicus (26/100), lugdunensis (18/100), aureus (15/100) and epidermidis (14/100) were dominant. S. aureus was 100% resistant to penicillin and sulfamethoxazole/trimethoprim. Vancomycin-resistant S. aureus showed intermediate resistance (51%), which might indicate the dissemination of vancomycin resistance in the community and imply food safety hazards. The percentage of resistance to ß-lactams was variable, with the highest resistance being to penicillin (94%) and lowest to ampicillin-sulbactam (22%). Antimicrobial resistance was mainly against penicillin (94%), clindamycin (90%) and sulfamethoxazole/trimethoprim (82%). The results indicate that chicken and beef raw meat are an important source of antibiotic-resistant CPS and CNS.