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.

Poxvirus infections of avian species worldwide are caused by viruses of a single genus (Avipoxvirus), they belong family Poxviridae. The turkey pox virus (TKPV) is one of the most serious illnesses and a major source of financial losses for the developing turkey industry. Even though the fowl pox virus vaccine is used to prevent the disease, TKPV still creates significant economic difficulties, therefore isolating and adapting the virus is crucial for the development of homologous vaccines that will manage the illness. In the summer of 2021, the TKPV disease spread to numerous farms inside the Giza Governorate. Ten samples of skin lesions from various farms were gathered and given labels. In accordance with Koch's postulates, the collected samples were ground, homogenized, and centrifuged to extract supernatant, which was then injected into healthy birds to confirm the disease symptoms and susceptibility. For more confirmation the virus was isolated on embryonated chicken egg (ECE) through chorio-allantoic membrane (CAM) route at 10 day old after 5 to 6 days of inoculation the pock lesion appeared  A fragment of the P4b gene coding for DNA polymerase core protein of TKPV was amplified by PCR then sequenced. A newly discovered isolate of field-isolated TKPV was revealed by phylogenetic analysis to have high similarity (between 96% and 100%) with the sequences that have been published. The new isolate of Turkey pox virus strain MKP 334 core protein (P4b) gene (with Accession No. MZ983434.1 in GenBank) has 100% to MG787222.1 Turkey pox virus isolate TurPVIR12 p4b gene and differ from the only isolated strain of TKPV that isolated at 2020 in Mansoura (MT219997.1 Turkey pox virus strain mans17T P4b 2020 EGYPT) but they have 99.6 identity.

FowlPox virus (FPV) was detected in eight chickens suffering from a diphtheritic lesion on the oropharynx and trachea with nodular skin lesions around the unfeathered parts in two Egyptian governorates (El-Sharkia and Ismailia governorates) during summer 2023. A variety of serological and molecular methods were performed for identification and characterization of the virus. on specific-pathogen free (SPF) embryonated chicken eggs via chorio-allantoic membranes (CAM), the distinguishing focal pock lesions were detected on CAM. Concerning electron microscopy, FPV appeared as enveloped quadrangular brick shaped Avipoxvirions. The neutralizing antibodies level against FPV were detected in all eight samples. Serum neutralization test showed a neutralization index of ≥ 1.6 in all serum samples, meanwhile ELISA test displayed an S/P ratio of ≥ 1.5 in the affected chickens. Notably, two positive FPV samples were sequenced then submitted to the GenBank (Sharquia-1 and Ismilia-2 with accession numbers; OR920788-OR920789). The phylogenetic tree construction based on the fpv167- (P4b) gene of FPV revealed high nucleotide identity with Elsharqyia_FWPV1, Elsharqyia_FWPV2 and Fowlpox-AN5, FWPVN, FWPVD (Egyptian isolates) with nucleotide identity percentage 100%, 99%, 100%, 99%,99%; respectively. Likewise, FPV isolates were of low homology with VSVRI-Vac (vaccinal strain) with 88% similarity. In context, the local recent our strains can be applied in vaccine production for appropriate vaccination programs in Egypt.

The aim of this study was to examine viruses associated with gastrointestinal illnesses in ducks collected from four governorates in Egypt (Sharkia, Gharbia, Dakahlia, and Qaliobia) during 2021-2022. These ducks underwent comprehensive clinical examinations and post-mortem analyses. All the flocks exhibited various forms of diarrhea. Additionally, 59.5% of the duck flocks manifested respiratory symptoms, while 57 % showed uneven growth, locomotory dysfunction (42.8%). Post-mortem findings consistently included enteritis in all examined flocks. To directly identify viruses associated with digestive illnesses, 42 aseptic intestine samples were obtained from recently deceased or sacrificed ducks. PCR analysis identified four positive samples out of the 42 (9.5%), with one containing Avian Rotavirus (AvRV) and three carrying Muscovy Duck Parvovirus (MDPV). Additionally, histopathological examination of the liver and intestine from PCR-positive flocks showed findings consistent with those typically observed in MDPV infections. This study concluded that the primary viruses associated with digestive illnesses in Egyptian ducks are MDPV and AvRV. Importantly, this research represents the first-ever detection of Rotavirus in ducks in Egypt.

Poultry bacterial pathogen is a major problem in poultry farms, with serious consequences for poultry and human. Two hundred samples of apparently health and freshly died broiler were collected from different commercial farms at Sharkia governorate, Egypt. Salmonella spp. detected in (29%) of examined specimens and serologically identified into S. Typhimurium, S. Kentucky, S. Infantis, S. Enteritidis and S. Agama with percentages 9.5%, 6.5%, 5%, 4.5% and 3.5%, respectively. Meanwhile, Pseudomonas spp. detected in (19%), the most prevalent serotype was P. aeruginosa O2, O5, O9, and O11. However, the antimicrobial-resistant strains of pathogens continuously emerge, with ineffective of medical treatments, thus, the isolates were examined for detection of multidrug resistant (MDR), doxycycline exhibited the highest antimicrobial activity against Salmonella spp. (55.17%); and ceftriaxone and doxycycline against Pseudomonas spp. (52.63%). Uniplex PCR examination for ampC, stn, tetA(B), integrase genes on MGEs were detected in all Salmonella spp. isolates, and mexR, tetA (B), integrase genes in all examined MDR P. aeruginosa isolates, meanwhile, exoU detected in 80% on MGEs. A novel antibacterial strategy was achieved to minimize economic burdens and the health associated with antimicrobial resistance which obliterate pathogens without any adverse effects on poultry and human. Therefore, the application of a trial using M. Oleifera nanoemulsion in order to control the multidrug resistant genes expression. These findings demonstrated that M. oleifera nanoemulsion was a good choice to its potential as a drug that can be used against Salmonella and P. aeruginosa in poultry industry.

Heartwater, one of the major tick-borne diseases of some domestic and wild ruminants in Africa, is caused by Ehrlichia ruminantium. The genetic diversity of E. ruminantium isolates renders the available vaccine ineffective against certain virulent isolates. To better understand the E. ruminantium genotypes in South Africa, a total of 1004 Amblyomma hebraeum tick deoxyribonucleic acid (DNA) samples from cattle in three South African provinces were tested by pCS20 Sol1 real-time polymerase chain reaction (qPCR) and characterised by multilocus sequence typing (MLST) using five housekeeping genes. Out of 1004 samples tested, 222 (22%) were positive for E. ruminantium. The occurrence of E. ruminantium in Mpumalanga, KwaZulu-Natal and Limpopo provinces was 19%, 22% and 27%, respectively. The E. ruminantium positive samples were screened for housekeeping genes and sequenced. Phylogenetic analysis revealed three main lineages: clade 1 made up of worldwide isolates (eastern, southern Africa, and Caribbean isolates), clade 2 comprised only West African isolates and clade 3 consisted of Omatjenne, Kümm2 and Riverside. Some study sample sequences were not identical to any of the reference isolates. However, they could all be grouped into the worldwide clade. Genetic variation in the sequenced regions was observed in the form of single nucleotide polymorphisms (SNPs). Using MLST to characterise E. ruminantium field isolates allowed the South African genotypes to be clearly distinguished from the distinct West African isolates. Contribution: Characterisation of E. ruminantium field isolates is important for the control of heartwater and contributes to preliminary knowledge required for the development of a more practical vaccine against heartwater.

This study targeted isolation and characterization of potential bacteriophages (phages) infecting MDR pathogenic bacteria recovered from chickens and analyzed their efficacy as bio-control agents. A total of 45 different bacterial isolates (18 E. coli, 16 Salmonellae spp., 5 Staphylococcus spp., 2 Pseudomonas spp., 1 Proteus mirbalis, 1 Citrobacter spp., 1 Enterobacter aerogenes and 1 Klebsiella pneumonia) were obtained from chickens in the current study and previous studies. The identified isolates were investigated for the presence of virulence genes and MDR using PCR and disc diffusion method, respectively. Nine purified phages classified morphologically into 3 families (Myoviridae, Siphoviridae and Podoviridae) using Transmission Electron Microscope were recovered from chicken intestinal contents and showed viability at wide pH range, resistance to organic solvents and thermostability at high temperatures (up to 80ºC). The potential phages exhibited various bacterial host ranges using the spot test and the efficiency of plating (EOP) assay. The results revealed the prevalent of pathogenic E. coli and salmonella serovars among the recovered isolates with different virulence and genotypic patterns. The lytic phages were highly stable and have the capacity to infect different pathogenic MDR bacterial strains. This study demonstrated that these promising phages of avian origin could be used to control the pathogenic MDR E. coli and Salmonella serovars which possess public concerns on human health and poultry industry.

FADV has caused high economic losses in poultry industry in Egypt in the last few years. The study aimed to detect and genetically characterize the fowl adenovirus (FAdV) species prevalent in Egyptian commercial broiler chicken flocks during 2023. The 63 suspected samples were collected from Egyptian broiler chickens from 5 governorates during 2023. The molecular characterization was performed by using polymerase chain reaction (PCR) and the positive samples was isolated in primary chicken embryo liver (CEL) cells. The genetic characterization of 8 selected samples represented different governorates by sequencing of loop 1 (L1) of the hexon gene. Clinically, the poultry suffered from depression, watery diarrhea, and ascites and decreased body weight with a mortality rate of 10–30%. The post-mortem inspection showed liver was pale, enlarged with petechial haemorrhage. 27 out of 63 samples (42.8%) were positive by PCR. The molecular charctersation of the L1 hexon gene’s revealed that the FADV (from Eg-ANY1-2023 to EG-ANY4-2023) genetically charcterized as FADV-D 2/11 strains, the FADV-EG-ANY5-2023 to FADV-EG-ANY8-2023 genetically characterized as FADV E/8a and FADV E/8b. By mutation analysis, the strains in our study related to FADV-E/8a (FADV-EG-ANY5, ANY6) had R171K in the HVR4 and strain related to 8b (FADV-EG-ANY6, ANY7) had S95N in the HVR2 and A91T between HVR1 and HVR2 compared to other reference strains. Thus, these findings demonstrate that many mutated virus genotypes are circulating in commercial chicken flocks. Further research is needed to study the pathogencity of these strains and implement control measures and vaccine production to prevent economic loss in the poultry industry.

Equine herpes virus-1 (EHV-1) is a major cause of economic losses in horse industry all over the world especially in the Arabian countries. The aim of the study is the isolation and identification of the local equine herpesvirus-1 at northern Egypt Governorates (Cairo, Dakahlyia, Qalyubia) during 2021- 2022. A total of 50 tissue organs were collected from aborted fetuses of affected mares recently after abortion. The collected samples were screened using consensus PCR for detection of equine herpesviruses (EHVs) using specific primers targeting DNA polymerase gene which revealed that 26 out of 50 samples were positive with percentage of 52%.The molecular positive samples were subjected to conventional PCR for further molecular identification and typing of EHVs using specific primers targeting glycoprotein (gB) gene which confirmed that all amplified products were for EHV-1. Attempts for isolation of EHV-1 were performed through propagation of molecular identified samples on Madin-Darby bovine kidney (MDBK) cell line which revealed that 19 samples developed characteristic cytopathic effect in form of cell rounding and rapidly enlarging grapes like cell aggregation, and then detached of the cell, while 13 samples induced pock lesions on CAMs of SPF-ECE after inoculation for three successive passages. Serological identification of EHV-1 isolates was confirmed by immunofluorescent antibody test (IFAT) which showed that 19 samples induced intracytoplasmic greenish yellow fluorescence. Selected EHV-1 isolate was subjected for transmission electron microscopy (TEM) for demonstration of virus morphology. The obtained results confirmed that EHV-1 is the major cause of abortion among Arabian mares which disrupts breeding and showcasing events. Future considerations about management strategies together with regular and effective vaccine trials are recommended for control of EHV-1.

Fasciolosis is a disease caused by the genus Fasciola which attacks wild animals, livestock and humans. Molecular characterization of Fasciola sp. important for identifying the species Fasciola sp. The aim of this research was to analyze the characteristics of the worm Fasciola sp. molecularly on samples of cattle slaughtered at the Kediri City Slaughterhouse Regional Technical Implementation Unit. This research was carried out from January to February 2023. The samples were adult worms Fasciola sp. taken from one beef cattle and one dairy cattle. The PCR used is conventional PCR with primers from mitochondrial DNA genes. PCR products are passed to the sequencing stage. The PCR test results read on 2% agarose gel electrophoresis showed that the PCR product had high specificity, namely forming a single band at position 752 bp showed positive for F. gigantica. the results of the phylogenetic tree of Fasciola sp. with data in GenBank showing Fasciola sp. beef and dairy cattle isolates in Kediri City are closely related to Fasciola sp. dairy cattle isolate from China with Accession Number KF543343.1 and Fasciola sp. long-haired cattle (yak) isolate from China Accession Number MH621335.1. The existence of international livestock trade can result in the introduction of various diseases, one of which is Fasciolosis. Livestock import-export activities from China allow the introduction of Fasciola sp. to Indonesia.