Prevalence, genetic diversity, and antimicrobial susceptibility of Vibrio spp. infected gilthead sea breams from coastal farms at Damietta, Egypt.

BACKGROUND: Vibriosis is one of the most serious bacterial diseases and causes high morbidity and mortality among cultured sea breams. This study was undertaken to track the surveillance of Vibrio infection and its correlation to environmental factors. A total of 115 gilthead sea breams were collected seasonally from a private earthen pond fish farm in the Shatta area of Damietta, Egypt from September 2022 to July 2023. Physicochemical parameters of water were analyzed, and heavy metal levels were measured. The fish samples were subjected to clinical, bacteriological, Enterobacterial Repetitive Intergenic Consensus (ERIC) fingerprinting, and hematoxylin and Eosin histopathological staining. RESULTS: The results revealed significant variations in the water quality parameters over different seasons, in addition to an increase in heavy metals. Naturally infected fish showed external signs and postmortem lesions that were relevant to bacterial infection. Two dominant Vibrio subspecies of bacteria were identified: V. alginolyticus (205 isolates) and V. fluvialis (87 isolates). PCR confirmed the presence of V. alginolyticus using the species-specific primer collagenase at 737 bp. The highest prevalence of V. alginolyticus was detected during the summer season (57.72%), and the lowest prevalence was observed in autumn (39.75%). The correlation analysis revealed a positive relationship between V. alginolyticus and water temperature (r = 0.69). On the other hand, V. fluvialis showed a high prevalence during the autumn season (25.30%) and the lowest prevalence during the summer season (10.56%), where it was negatively correlated with water temperatures (r =-0.03). ERIC fingerprinting showed genetic variation within the Vibrio isolates. Antimicrobial susceptibility testing revealed sensitivity to ciprofloxacin and doxycycline, and resistance to amoxicillin and erythromycin. The multiple antibiotic resistance (MAR) index values for V. alginolyticus and V. fluvialis ranged from 0.3 to 0.7, with a multi-drug resistance pattern to at least three antibiotics. Histopathological alterations in the affected tissues revealed marked hemorrhage, vascular congestion, and hemosiderosis infiltration. CONCLUSION: This study provides insights into the potential propagation of waterborne diseases and antibiotic resistance in the environment. Ensuring that the environment does not serve as a reservoir for virulent and contagious Vibrio species is a critical concern for regional aquaculture industries. Therefore, we recommend implementing environmental context-specific monitoring and surveillance tools for microbial resistance.

Technological solutions to landfill management: Towards recovery of biomethane and carbon neutrality.

Inadequate landfill management poses risks to the environment and human health, necessitating action. Poorly designed and operated landfills release harmful gases, contaminate water, and deplete resources. Aligning landfill management with the Sustainable Development Goals (SDGs) reveals its crucial role in achieving various targets. Urgent transformation of landfill practices is necessary to address challenges like climate change, carbon neutrality, food security, and resource recovery. The scientific community recognizes landfill management's impact on climate change, evidenced by in over 191 published articles (1998-2023). This article presents emerging solutions for sustainable landfill management, including physico-chemical, oxidation, and biological treatments. Each technology is evaluated for practical applications. The article emphasizes landfill management's global significance in pursuing carbon neutrality, prioritizing resource recovery over end-of-pipe treatments. It is important to note that minimizing water, chemical, and energy inputs in nutrient recovery is crucial for achieving carbon neutrality by 2050. Water reuse, energy recovery, and material selection during manufacturing are vital. The potential of water technologies for recovering macro-nutrients from landfill leachate is explored, considering feasibility factors. Integrated waste management approaches, such as recycling and composting, reduce waste and minimize environmental impact. It is conclusively evident that the water technologies not only facilitate the purification of leachate but also enable the recovery of valuable substances such as ammonium, heavy metals, nutrients, and salts. This recovery process holds economic benefits, while the conversion of CH4 and hydrogen into bioenergy and power generation through microbial fuel cells further enhances its potential. Future research should focus on sustainable and cost-effective treatment technologies for landfill leachate. Improving landfill management can mitigate the adverse environmental and health effects of inadequate waste disposal.

Tuning cationic/anionic dyes sorption from aqueous solution onto green algal biomass for biohydrogen production.

Global water security and energy demands associated with uncontrollable population growth and rapid industrial progress are one of the utmost serious needs dangerously confronting humanity. On account of waste as a wealth strategy; a multifunctional eco-friendly sorbent (MGAP) from green alga was prepared successfully for remediation of cationic/anionic organic dyes and biohydrogen production. The structural and morphological properties of sorbent were systematically scrutinized by a variety of spectral analyses. The loading capacity of MGAP towards rhodamine B (RhB) and methyl orange (MO) dyes was inclusivity inspected under variable experimental conditions. The adsorption kinetics of both dyes onto MGAP was in good agreement with pseudo-second-order theory, whereas adsorption isotherms could fit well with the Langmuir model, with satisfactory loading capacities of 144.92 and 196.04 mg g-1 for RhB and MO molecules, respectively. Moreover, ultra-sonication treatment admirably decreased the sorption equilibrium time from 180.0 min to 30.0 min. Furthermore, spent sorbent was managed particularly for biohydrogen production with a measured yield of 112.89, 116.59, and 128.17 mL-H2/gVS for MGAP, MGAP-MO, and MGAP-RhB, respectively. Overall, the produced MGAP can potentially be offered up as a promising dye scavenger for wastewater remediation and biohydrogen production, thereby fulfilling waste management and circular economy.

Microarray-based detection of resistance genes in coagulase-negative staphylococci isolated from cattle and buffalo with mastitis in Egypt.

The present study aimed to provide a detailed characterization of coagulase-negative staphylococci (CoNS) isolated from cows and buffaloes with mastitis. The study included seventy-five CoNS isolates (60 came from cattle and 15 from buffaloes) originating from 68 individual quarters of 67 dairy cows (53 cattle and 14 buffaloes). The animals belonged to five different small holding dairy herds (n = 140 cows) that show clinical or subclinical mastitis. CoNS isolates were phenotypically characterized using MALDI-TOF-MS and were further genotypically characterized by microarray-based assays. Furthermore, the antimicrobial susceptibility of CoNS strains which carried the mecA gene was examined by broth microdilution. The occurrence of CoNS in the respective five herds was 10.5%, 14.7%, 14.8%, 12.8%, and 9.9%, with an average of 12.4%. Six different CoNS species were identified: S. sciuri (n = 37; 30 from cattle and 7 from buffaloes), S. chromogenes (n = 14; 8 from cattle and 6 from buffaloes), S. haemolyticus (n = 10; nine from cattle and one buffalo), S. xylosus (n = 10; nine from cattle and one buffalo), S. hyicus (n = 2), S. warneri (n = 1), and unidentified CoNS (n = 1). Twenty percent (20%) of CoNS isolates (17.3% of cattle origin) carried at least one antimicrobial resistance gene, while 4% of the isolate including two isolates of S. haemolyticus and one S. warneri of cattle origin carried the mecA gene and were phenotypically identified as methicillin-resistant strains. The genes detected were blaZ (16%), followed by tet(K) (8%), aacA-aphD (4%), aphA3 (2.6%), msr(A) (2.6%), [far1 (2.6%), and fusC (2.6%)], sat (2.6%), and cat (1.3%) conferring resistance to penicillin, tetracycline, gentamicin, neomycin/kanamycin, erythromycin, fusidic acid, streptothricin, and chloramphenicol, respectively. The majority of investigated CoNS strains displayed considerably low prevalence of resistance genes, while resistance to more than three antibiotics was found in S. haemolyticus and S. warneri. Implementing effective preventive measures is, therefore, important for limiting the transmission of CoNS, rather than using antibiotics to control mastitis in bovines.

Staphylococci in cattle and buffaloes with mastitis in Dakahlia Governorate, Egypt.

The aim of this study was to provide the first detailed insight into the population structure of Staphylococcus aureus in one modern dairy farm (Gamasa) and several household cows and buffaloes in Dakahlia Governorate, Egypt. Eight hundred seventy-two quarter milk samples of 218 dairy cattle and buffaloes with clinical and subclinical mastitis were investigated. Bacteria were identified using matrix-assisted laser desorption/ionization time-of-flight mass spectrometry, and staphylococci were further characterized by DNA sequencing of 16S rRNA genes and microarray analysis. Staphylococcus aureus was present in 5.6% of all collected samples, whereas methicillin-resistant S. aureus (MRSA) represented 24.5% of all identified S. aureus (12/49). Six clonal complexes (CC) of S. aureus were detected. Staphylococcus aureus CC398 (ST291/813)-MSSA (methicillin-susceptible S. aureus) was identified frequently in the Gamasa farm in addition to a few CC5-MRSA-V isolates. However, a small number of different isolates of S. aureus were found in household cattle and buffaloes harboring different CC. The presence of these genotypes of S. aureus in milk might indicate a public health hazard, because all of these CC have previously been isolated from human patients. Thus, a recommendation was given to the owner of the dairy farm to review the hygiene regimen on the farm. In perspective, further investigation regarding S. aureus screening of all lactating cows and personnel on the farm is warranted.

Effect of organic loading rates on the performance of membrane bioreactor for wastewater treatment behaviours, fouling, and economic cost.

Although submerged membrane bioreactor (MBR) are widely used in treating municipal wastewater and recovery of potential resources, membrane operational parameters and membrane fouling control remain debated issues. In this study, the treatment of municipal wastewater by MBR at high-biomass sludge (MLSS (g/L) ranging from 5.4 g/L to 16.1 g/L) was assessed at an organic loading rates (OLRs) ranging from 0.86 to 3.7 kg COD/m3d. The correlation between trans-membrane pressure and total fouling resistance was thoroughly investigated in this study. According to the findings, greater OLRs of 0.86 to 3.7 kg COD/m3d caused a decrease in COD, BOD, and NH4-N removal efficiency, and higher OLRs of 3.7 kg COD/m3d resulted in a higher increase in total fouling resistance (Rt). The economic study of using the MBR system proved that for a designed flow rate of 20 m3/d, the payback period from using the treated wastewater will be 7.98 years, which confirms the economic benefits of using this MBR for treating municipal wastewater. In general, understanding the challenges facing the efficiency of MBR would improve its performance and, consequently, the sustainability of wastewater reclamation.

Mass flow and consumption calculations of pharmaceuticals in sewage treatment plant with emphasis on the fate and risk quotient assessment.

In Egypt, pharmaceuticals consumption increased dramatically owing to the population growth and the unrestricted sale manner. Accordingly, the occurrence and fate of nine common pharmaceutical active compounds (PhACs) were scrutinized at a sewage treatment plant (STP) in Giza, Egypt. The levels of these PhACs were assessed in different the wastewater treatment stages and dewatered sludge phase using high-performance liquid chromatography coupled with photodiode arrays detector. The average concentrations of the total PhACs detected in influent, primary sedimentation effluent (PSE) and final effluent (FE) were 227, 155 and 89 µg L-1, respectively. The overall removal efficiency of the individual PhACs ranged from 18 to 72% removal. The occurrence trend revealed that biodegradation and adsorption are the concurrently removal mechanisms of the studied PhACs. The overall consumption per day in West of Greater Cairo was estimated based on influent concentration of STP. Sulfamethoxazole, paracetamol and diclofenac were detected with the highest levels in the influent of STP, PSE and FE as well as in the dewatered sludge. Furthermore, the high concentrations of these compounds in the sludge confirm the adsorption pathway removal of theses PhACs. The risk quotient (RQ) assessment for the detected PhACs in FE is greatly higher than the predicted non-effect concentration (PNEC). Conclusively, the FE of STP is considered a risky source for PhACs in adjacent surface water.

Prevalence and Morphological Investigation of Parasitic Infection in Freshwater Fish (Nile Tilapia) from Upper Egypt.

Fish are a source of high-quality protein with low cholesterol, but they are susceptible to parasitic infections, which have a significant impact on aquaculture, in addition to their zoonotic potential. The present study estimated parasitic infections and evaluated the diversity of zoonotic parasites in freshwater Nile tilapia (Oreochromis niloticus) in Assiut Governorate, Upper Egypt. A total of 300 samples were randomly collected from the Assiut Governorate. These fish were examined for both ectoparasites and endoparasites, followed by the experimental infection of mice with encysted metacercariae (EMC) for the retrieval of the adult worms. The overall prevalence of the variable parasites was 82% (246 of 300). Both ecto- and endoparasites were detected in 41% (123 of 300) of the examined fish. The identified ectoparasites were Gyrodactylus, Dactylogrus, Cichlidogyrus, Trichodina and Icthyophthirius multifiliis, in 5%, 4%, 22%, 6% and 4% of the fish, respectively. The endoparasites were trematodes (Orientocreadium batrachoides 3%), nematodes (Contracaecum. 2%), acanthocephala (Acanthosentis tilapiae 25%) and protozoa that included Isospora and Eimeria spp., in 1% and 8% of fish, respectively. Myxobolus was detected in 2% of the examined fish. The overall prevalence of encysted metacercariae (EMC) was 95% (285 of 300), while infection with macroscopic EMC had a prevalence of 37% and microscopic EMC had a prevalence of 58%. The adult worms recovered from the experimental infections were Prohemistomum vivax and Mesostephanus spp., which belong to the family Cyathocotylidae. Collectively, these findings reflect the relatively high occurrence of parasites among the studied fish, confirming the necessity of strict measures to control infection.

Management of wastewater from the vegetable dehydration industry in Egypt--a case study.

Management of wastewater from the vegetable dehydration industry was the subject of this study. A continuous monitoring programme for wastewater was carried out for almost four months. The characterization of the wastewater indicated that the vegetable dehydration wastewater contains moderate concentrations of organics, solids and nutrients. The wastewater was subjected to three different treatment processes, namely aerobic treatment, anaerobic treatment and chemical coagulation-flocculation treatment. For aerobic treatment, the removal of chemical oxygen demand (COD), biochemical oxygen demand (BOD5) and total suspended solids (TSS) was accomplished within 5 h, and no further reduction was observed after that, with the steady state COD and BOD5 removal efficiencies being 95% +/- 10% and 97% +/- 8%, respectively. For anaerobic treatment, the removal efficiencies for COD, BOD5 and TSS were 67-81%, 70-86% and 56-69%, respectively at hydraulic retention times (HRTs) of 5, 6 and 8 h. Chemical coagulation-flocculation treatment also achieved good results. The COD removal efficiency was 72%, 51% and 75% for ferric chloride (56 g/m3 of wastewater), lime (140 g/m3 of wastewater) and ferric chloride aided with lime (100 g/m3 for ferric chloride and 200 g/m3 for lime), respectively. The corresponding TSS removal values were 92% +/- 17%, 20% +/- 7% and 93% +/- 9%. Based on the available results and the seasonally operated mode of this industry in Egypt, the chemical coagulation-flocculation process is therefore considered to be moste applicable from a technical point of view and for the simplicity of operation and maintenance.

Integration of microbial electrochemical systems and photocatalysis for sustainable treatment of organic recalcitrant wastewaters: Main mechanisms, recent advances, and present prospects.

In recent years, microbial electrochemical systems (MESs) have demonstrated to be an environmentally friendly technology for wastewater treatment and simultaneous production of value-added products or energy. However, practical applications of MESs for the treatment of recalcitrant wastewater are limited by their low power output and slow rates of pollutant biodegradation. As a novel technology, hybrid MESs integrating biodegradation and photocatalysis have shown great potential to accelerate the degradation of bio-recalcitrant pollutants and increase the system output. In this review, we summarize recent advances of photo-assisted MESs for enhanced removal of recalcitrant pollutants, and present further discussion about the synergistic effect of biodegradation and photocatalysis. In addition, we analyse in detail different set-up configurations, discuss mechanisms of photo-enhanced extracellular electron transfer, and briefly present ongoing research cases. Finally, we highlight the current limitations and corresponding research gaps, and propose insights for future research.

Genetic Relatedness, Antibiotic Resistance, and Effect of Silver Nanoparticle on Biofilm Formation by Clostridium perfringens Isolated from Chickens, Pigeons, Camels, and Human Consumers.

In this study, we determined the prevalence and toxin types of antibiotic-resistant Clostridium perfringens in chicken, pigeons, camels, and humans. We investigated the inhibitory effects of AgNPs on biofilm formation ability of the isolates and the genetic relatedness of the isolates from various sources determined using RAPD-PCR. Fifty isolates were identified using PCR, and all the isolates were of type A. The cpe and cpb2 genes were detected in 12% and 56% of the isolates, respectively. The effect of AgNPs on biofilm production of six representative isolates indicated that at the highest concentration of AgNPs (100 µg/mL), the inhibition percentages were 80.8-82.8%. The RAPD-PCR patterns of the 50 C. perfringens isolates from various sources revealed 33 profiles and four clusters, and the discriminatory power of RAPD-PCR was high. Multidrug-resistant C. perfringens isolates are predominant in the study area. The inhibition of biofilm formation by C. perfringens isolates was dose-dependent, and RAPD-PCR is a promising method for studying the genetic relatedness between the isolates from various sources. This is the first report of AgNPs' anti-biofilm activity against C. perfringens from chickens, pigeons, camels, and humans, to the best of our knowledge.

Parasitological, Molecular, and Epidemiological Investigation of Cryptosporidium Infection Among Cattle and Buffalo Calves From Assiut Governorate, Upper Egypt: Current Status and Zoonotic Implications.

Details about the epidemiological patterns and real contributions of different reservoir animals in maintaining the transmission cycle of Cryptosporidium spp. in Upper Egypt remain lacking. This study was designed to investigate the occurrence of Cryptosporidium spp. in cattle and buffalo (n = 608) from Upper Egypt. The parasite for the resulting positive samples by fecal examination was molecularly identified using nested PCR targeting the small subunit rRNA. Moreover, several explanatory variables, including animals' age, sex, condition, seasonal variations, were examined to describe the epidemiological pattern of the disease. Interestingly, the fecal examination revealed that 33.55% (204/608) of the animals under study were infected with Cryptosporidium, including 38.27% among cattle and 28.16% among buffalo. The parasite was molecularly identified using nested PCR, and their amplicons were identified in almost all fecal samples using microscopy (202/204). According to age as an individual variable factor, the infection rates of Cryptosporidium spp. in cattle calves with ages of <1, 1-3, and >3 months were 39.13, 34.04, and 54.54%, respectively. Meanwhile, in buffalo calves, the occurrence rates were 28.57, 27.27, and 29.41%, respectively. Regarding sex, female cattle calves were more susceptible to Cryptosporidium infection (51.28%) than males (26.19%) (p < 0.05), whereas male buffalo calves had a higher infection rate (32.25%) than females (25%). According to seasonal variations, the infection rates of Cryptosporidium spp. in cattle calves during spring, summer, autumn, and winter were 42.11, 30.43, 30, and 52.63%, respectively. In contrast, lower infection rates of 30, 21.42, 23.52, and 35% were reported in buffalo calves during spring, summer, autumn, and winter, respectively. The rate of infection was 45.16% in diarrheic cattle calves and 15.78% in non-diarrheic ones (p < 0.05). Meanwhile, the infection rate was 33.96% in diarrheic buffalo calves and 11.11% in non-diarrheic ones (p < 0.05). This study reported a higher occurrence of Cryptosporidium infection among the animals under study and revealed that buffalos and cattle can contribute to maintaining the transmission cycle of this zoonotic parasite in Upper Egypt.

Acaricidal Properties of Four Neem Seed Extracts (Azadirachta indica) on the Camel Tick Hyalomma dromedarii (Acari: Ixodidae).

Tick infestation remains one of the major health problems that affect the productivity and comfort of camels. The control of ticks mainly relies on using chemical acaracides. Limited information is available on the potential benefits and activity of various neem extracts on Hyalomma ticks. The present study investigated the acaricidal activity of neem seed extracts at different concentrations against developmental stages of the camel tick Hyalomma dromedarii in comparison to Butox and diazinon. The acaricidal activity of three extracts, namely, hexane extract (HE), methyl chloride extract (MCE), and methanol extract (ME), of neem seeds (Azadirachta indica) were tested at varying concentrations of 5, 10, 15, and 20% on engorged H. dromedarii female ticks at days 1, 3, 5, 7, 12, 16, 20, 28, 37, and 43 after treatment (DPT). Interestingly, results of applying different neem seed extracts to engorged H. dromedarii female ticks showed that the most effective extract was hexane at concentration 20%, causing 100% mortality at 1st day post-application, while methanol extract at 20% and dichloromethane extract at 20% caused the death of all ticks at 28th day posttreatment as compared to Butox® 5.0 and Diazinon-60, which resulted in mortality of all ticks at 3 and 5 DPT, respectively. In addition, no mortality was reported with the application of aqueous extract (AE), which served as the control group. Furthermore, the neem hexane extract exhibited high efficacy against reproductive performance of female ticks, whereas no fertility or oviposition was reported at all of their concentrations. Additionally, no hatchability occurred using all neem extracts, except the aqueous extract, which showing no effect. In the present study, larvae responded more rapidly to the plant extracts, whereas mortality of all larvae was recorded at 24 h after treatment with 5% hexane. Taken together, this study pointed out that the acaricidal effect of hexane extract of neem seeds was more effective and could be economically used for controlling H. dromedarii ticks.

Increasing 2 -Bio- (H2 and CH4) production from food waste by combining two-stage anaerobic digestion and electrodialysis for continuous volatile fatty acids removal.

A novel approach of using two stage anaerobic digestion coupled with electrodialysis technology has been investigated. This approach was used to improving bio hydrogen and methane yields from food waste while simultaneously producing a green chemical feedstock. The first digester was used for hydrogen production and the second digester was used for methane production. The first digester was combined with continuous separation of volatile fatty acids using electrodialysis. The concentrations of carbohydrates, proteins and fats in the prepared food waste were 22.7%, 5.7% and 5.2% respectively. Continuous removal of volatile fatty acids during fermentation in the hydrogen digester not only increased hydrogen yields but also increased the production rate of volatile fatty acids. As a result of continuous VFA separation, hydrogen yields increased from 17.3 mL H2/g VS fermenter to 33.68 mL H2/g VS fermenter. Methane yields also increased from 28.94 mL CH4/g VS fermenter to 43.94 mL CH4/g VS fermenter. This represents a total increase in bio-energy yields of 77.1%. COD reduced by 73% after using two stage anaerobic digestion, however, this reduction increased to 86.7% after using electrodialysis technology for separation of volatile fatty acids. Electrodialysis technology coupled with anaerobic digestion improved substrate utilization, increased bioenergy yields and looks to be promising for treating complex wastes such as food waste.

Epidemiological, Morphometric, and Molecular Investigation of Cystic Echinococcosis in Camel and Cattle From Upper Egypt: Current Status and Zoonotic Implications.

Cystic echinococcosis has been considered one of the major parasitic zoonoses which is associated with severe economic losses. The present study was undertaken to investigate the occurrence, organ distribution, cyst fertility, and viability of cystic echinococcosis in slaughtered camels and cattle from various abattoirs in Assiut Governorate, Egypt. The work also involved morphological, morphometric, and molecular identification of the parasite. The occurrence of hydatid cysts was investigated in total number of 100 lungs of camels and 574 liver and lungs of cattle admitted to three slaughterhouses at Assiut Governorate, Egypt. Moreover, several individual variable factors, including organ involvement, age, sex, and hydatid cyst characteristics, were studied to identify their possible association with the occurrence of the disease. Genomic DNA was extracted from the hydatid cysts, followed by molecular identification of the parasite through amplification of ribosomal DNA internal transcribed spacer (ITS) regions. Hydatid cysts were found in 6 camels (6%) out of 100 inspected camels, while 5 hydatid cysts (0.87%) were detected in a total number of 574 cattle examined. The parasite was detected exclusively in lungs of camels, while lungs were the main organ infected by the parasite in cattle and one hydatid cyst was found in the liver (0.17%). In camel, 66.7, 16.65, and 16.65%of detected cysts were fertile, sterile, and calcified, respectively, while in cattle, these percentages were 60, 20, and 20%, respectively. None of the studied variable factors were significantly associated with the occurrence of the disease in camels, with the exception that all cysts were found in the lung. Conversely, we found a significant association (P < 0.05) between the age and sex of the slaughtered cattle and the occurrence of hydatid cysts. In this respect, the rate of infection was higher in female cattle and those cattle more than 5 years (P < 0.05). The morphological, morphometric, and molecular studies confirmed the presence of the parasite. Taken together, our results concluded that camels and cattle play a potential role in maintaining the transmission cycle of this zoonotic parasite.

Molecular Characterization of Velogenic Newcastle Disease Virus (Sub-Genotype VII.1.1) from Wild Birds, with Assessment of Its Pathogenicity in Susceptible Chickens.

Newcastle disease (ND) is considered to be one of the most economically significant avian viral diseases. It has a worldwide distribution and a continuous diversity of genotypes. Despite its limited zoonotic potential, Newcastle disease virus (NDV) outbreaks in Egypt occur frequently and result in serious economic losses in the poultry industry. In this study, we investigated and characterized NDV in wild cattle egrets and house sparrows. Fifty cattle egrets and fifty house sparrows were collected from the vicinity of chicken farms in Kafrelsheikh Governorate, Egypt, which has a history of NDV infection. Lung, spleen, and brain tissue samples were pooled from each bird and screened for NDV by real-time reverse transcriptase polymerase chain reaction (RRT-PCR) and reverse transcriptase polymerase chain reaction (RT-PCR) to amplify the 370 bp NDV F gene fragment. NDV was detected by RRT-PCR in 22 of 50 (44%) cattle egrets and 13 of 50 (26%) house sparrows, while the conventional RT-PCR detected NDV in 18 of 50 (36%) cattle egrets and 10 of 50 (20%) of house sparrows. Phylogenic analysis revealed that the NDV strains identified in the present study are closely related to other Egyptian class II, sub-genotype VII.1.1 NDV strains from GenBank, having 99.7-98.5% identity. The pathogenicity of the wild-bird-origin NDV sub-genotype VII.1.1 NDV strains were assessed by experimental inoculation of identified strains (KFS-Motobas-2, KFS-Elhamoul-1, and KFS-Elhamoul-3) in 28-day-old specific-pathogen-free (SPF) Cobb chickens. The clinical signs and post-mortem changes of velogenic NDV genotype VII (GVII) were observed in inoculated chickens 3 to 7 days post-inoculation, with 67.5-70% mortality rates. NDV was detected in all NDV-inoculated chickens by RRT-PCR and RT-PCR at 3, 7, and 10 days post-inoculation. The histopathological findings of the experimentally infected chickens showed marked pulmonary congestion and pneumonia associated with complete bronchial stenosis. The spleen showed histocytic cell proliferation with marked lymphoid depletion, while the brain had malacia and diffuse gliosis. These findings provide interesting data about the characterization of NDV in wild birds from Egypt and add to our understanding of their possible role in the transmission dynamics of the disease in Egypt. Further research is needed to explore the role of other species of wild birds in the epidemiology of this disease and to compare the strains circulating in wild birds with those found in poultry.

Contamination Pathways can Be Traced along the Poultry Processing Chain by Whole Genome Sequencing of Listeria innocua.

Foodborne infection with Listeria causes potentially life-threatening disease listeriosis. Listeria monocytogenes is widely recognized as the only species of public health concern, and the closely related species Listeria innocua is commonly used by the food industry as an indicator to identify environmental conditions that allow for presence, growth, and persistence of Listeria spp. in general. In our study, we analyze the occurrence of Listeria spp. in a farm-to-fork approach in a poultry production chain in Egypt and identify bacterial entry gates and transmission systems. Prevalence of Listeria innocua at the three production stages (farm, slaughterhouse, food products) ranged from 11% to 28%. The pathogenic species Listeria monocytogenes was not detected, and Listeria innocua strains under study did not show genetic virulence determinants. However, the close genetic relatedness of Listeria innocua isolates (maximum 63 SNP differences) indicated cross-contamination between all stages from farm to final food product. Based on these results, chicken can be seen as a natural source of Listeria. Last but not least, sanitary measures during production should be reassessed to prevent bacterial contamination from entering the food chain and to consequently prevent human listeriosis infections. For this purpose, surveillance must not be restricted to pathogenic species.

Antimicrobial resistance pattern and virulence profile of S. aureus isolated from household cattle and buffalo with mastitis in Egypt.

Methicillin resistant S. aureus from cows with mastitis has received a growing interest worldwide. The present study aimed to provide a detailed description of the resistance and virulence traits of isolates from bovine mastitis samples. A total of 550 quarter milk samples were collected from 140 mastitic household dairy cows and buffalo from five herds at Dakahlia Governorate, Egypt, during 2017 and 2018. Staphylococcus spp. were isolated and differentiated using MALDI-TOF MS. A genotypic characterization was performed for S. aureus isolates using DNA-microarray and staphylococcal protein A (spa) typing. Furthermore, antibiotic resistances were phenotypically confirmed using broth microdilution. Six different clonal lineages (CC1-MRSA, CC5-MRSA, CC45-MRSA, CC97-MSSA, CC50-MSSA and CC1153-MSSA), including seven spa types (t127, t688, t132, t267, t521, t224 and t903) were identified. Spa type t267 was the most dominant among the investigated herds. This is the first report of the occurrence of clonal lineages CC97, CC1, CC45, CC50 and CC1153 from bovine mastitis in Egypt. All MRSA isolates and 33.3 % of MSSA were multi-resistant (i.e. resistant to more than three classes of compounds). Various virulence determinants were also observed including leukocidins, hemolysins, and enterotoxins. The study demonstrates a low diversity of S. aureus isolates recovered from several dairy herds. The findings of the observed virulotypes can be useful for future studies on anti-virulence therapies, immunogenicity and vaccine development.

New Insights Into Potential Benefits of Bioactive Compounds of Bee Products on COVID-19: A Review and Assessment of Recent Research.

The recent emergence of COVID-19 represents one of the biggest challenges facing the world today. Despite the recent attempts to understand the epidemiological pattern and pathogenesis of the disease, detailed data about the physiology and pathology of the disease is still out of reach. Moreover, the lack of a widespread vaccine prompts an urgent call for developing a proper intervention strategy against the virus. Importantly, identification of novel molecules that target replication of the virus represents one of the promising strategies for the control this pandemic crisis. Among others, honey bee products contain numerous bioactive compounds such as propolis and several phenolic compounds that possess a wide range of therapeutic properties for combating various pathological disorders and infectious agents. The intention of the present review is to highlight the stages of SARS-CoV-2 lifecycle, the molecular mechanisms explaining the health benefits of honey bee products on COVID-19 physiology and pathology and the possible limitations. Further future research is suggested to explore more about bee natural bioactive compounds as potential candidates against SARS-CoV-2.

Pathological and Molecular Characterization of H5 Avian Influenza Virus in Poultry Flocks from Egypt over a Ten-Year Period (2009-2019).

Avian influenza virus (AIV) remains one of the enzootic zoonotic diseases that challenges the poultry industry in Egypt. In the present study, a total of 500 tissue samples were collected from 100 chicken farms (broilers and layers) suspected to be infected with AIV through the period from 2009 to 2019 from Dakahlia governorate, Egypt. These samples were pooled in 100 working samples and screened for AIV then the positive samples were subjected to histopathological examination combined with real time-polymerase chain reaction (RRT-PCR). RRT-PCR positive samples were also subjected to conventional reverse transcriptase-polymerase chain reaction (RT-PCR) for detection of H5 AIV and some of these resulting positive samples were sequenced for detection of the molecular nature of the studied virus. Interestingly, the histopathological examination revealed necrotic liver with leukocytic infiltration with degenerative changes with necrotic pancreatitis, edema, and intense lymphoid depletion of splenic tissue and hyperplastic tracheal epithelium. Likewise, edema and congested sub mucosal blood vessels and intense bronchial necrosis with hyalinized wall vascular wall and heterophils infiltration were reported. Pneumonic areas with intense leukocytic aggregation mainly and vasculitis of the pulmonary blood vessels were also detected in lung. Collectively, these significant pathological changes in examined tissues cohered with AIV infection. Regarding the molecular characterization, 66 samples were positive for AIV by RRT-PCR and 52 of them were positive for H5 AIV by RT-PCR. The phylogenetic analysis revealed that the H5 viruses identified in this study were aligned with other Egyptian H5N1 AIVs in the Egyptian sub clade 2.2.1, while some of the identified strains were aligned with other Egyptian H5N8 strains in the new Egyptian sub clade 2.3.4.4. Taken together, our present findings emphasize the wide spread of AIV in Egypt and the importance of developing an efficient surveillance and periodical screening program for controlling such disease of public health concern.