Bhattacharya, K., I. M. Shamkh, M. S. Khan, M. M. Lotfy, J. B. Nzeyimana, R. F. Abutayeh, N. M. Hamdy, D. Hamza, N. R. Chanu, P. Khanal, et al., "Multi-Epitope Vaccine Design against Monkeypox Virus via Reverse Vaccinology Method Exploiting Immunoinformatic and Bioinformatic Approaches.", Vaccines, vol. 10, issue 12, 2022. Abstract

(1) Background: The monkeypox virus is a zoonotic orthopox DNA virus that is closely linked to the virus. In light of the growing concern about this virus, the current research set out to use bioinformatics and immunoinformatics to develop a potential vaccine against the virus. (2) Methods: A multiepitope vaccine was constructed from the B-cell and T-cell epitopes of the MPXVgp181 strain using adjuvant and different linkers. The constructed vaccine was predicted for antigenicity, allergenicity, toxicity, and population coverage. In silico immune simulation studies were also carried out. Expression analysis and cloning of the constructed vaccine was carried out in the pET-28a(+) vector using snapgene. (3) Results: The constructed vaccine was predicted to be antigenic, non-allergenic, and non-toxic. It was predicted to have excellent global population coverage and produced satisfactory immune response. The in silico expression and cloning studies were successful in , which makes the vaccine construct suitable for mass production in the pharmaceutical industry. (4) Conclusion: The constructed vaccine is based on the B-cell and T-cell epitopes obtained from the MPXVgp181 strain. This research can be useful in developing a vaccine to combat the monkeypox virus globally after performing in-depth in vitro and in vivo studies.

Mohammed, R., S. M. Nader, D. A. Hamza, and M. A. Sabry, "Horse: a potential source of Cryptococcus neoformans and Cryptococcus gattii in Egypt.", BMC veterinary research, vol. 18, issue 1, pp. 17, 2022. Abstract

BACKGROUND: Cryptococcosis is an opportunistic mycozoonosis of global significance in a wide variety of host species. In equines, cryptococcosis is uncommon, and sporadic cases have been reported with rhinitis, sinusitis, pneumonia, and meningitis. Cryptococcus spp. represents a potential risk for immunosuppressed and healthy persons. In Egypt, epidemiological data on cryptococcal infection in horses are limited. The current study was carried out to investigate the occurrence of Cryptococcus spp. in horses and its possible role in the epidemiology of such disease in Egypt. A total of 223 samples was collected from different localities in Egypt included 183 nasal swabs from horses, 28 nasal swabs from humans, and 12 soil samples. Bacteriological examination and the identification of Cryptococcus spp. were performed. Molecular serotyping of Cryptococcus spp. was determined by multiplex PCR using CNa-70S/A-CNb-49S/A. The virulence genes (LAC1, CAP59, and PLB1) of the identified isolates were detected by PCR. Moreover, sequencing and phylogenetic analysis of the C. gattii gene from horses, humans, and soil isolates found nearby were performed.

RESULT: The overall occurrence of Cryptococcus spp. in horses were 9.3, 25, and 10.7% in horses, the soil, and humans, respectively. Molecular serotyping of the Cryptococcus spp. isolates recovered from the nasal passages of horses proved that C. gattii (B), C. neoformans, and two hybrids between C. neoformans (A) and C. gattii (B) were identified. Meanwhile, in case of soil samples, the isolates were identified as C. gattii (B). The human isolates were serotyped as C. gattii in two isolates and C. neoformans in only one isolate. Molecular detection of some virulence genes (LAC1), (CAP59), and (PLB1) were identified in both C. gattii and C. neoformans isolates. The C. gattii gene amplicons of the isolates from horses, humans, and the soil were closely related.

CONCLUSION: This study provides the first insights into the Egyptian horse ecology of Cryptococcus species and highlights the role of horses as asymptomatic carriers in disseminating the potentially pathogenic Cryptococcus spp. It also presents the possible risk of cryptococcosis infection in humans.

Elkazzaz, M., A. Ahmed, Y. E. - E. Abo-Amer, T. Hydara, A. Haikal, D. A. E. N. Razek, W. A. Eltayb, X. Wang, T. M. Karpiński, D. Hamza, et al., "In Silico Discovery of GPCRs and GnRHRs as Novel Binding Receptors of SARS-CoV-2 Spike Protein Could Explain Neuroendocrine Disorders in COVID-19.", Vaccines, vol. 10, issue 9, 2022. Abstract

Despite the intense research work since the beginning of the pandemic, the pathogenesis of COVID-19 is not yet clearly understood. The previous mechanism of COVID-19, based on ACE2 tropism and explained through a single receptor, is insufficient to explain the pathogenesis due to the absence of angiotensin-converting enzyme 2 (ACE2) receptors in most of the affected organs. In the current study, we used the PatchDock server to run a molecular docking study of both the gonadotropin-releasing hormone receptor (GnRHR) and G-protein-coupled-receptor (GPCR) with the SARS-CoV-2 spike protein. Molecular Dynamics (MD) simulations were run to analyze the stability of the complexes using the GROMACS package. The docking results showed a high affinity between the spike protein with the GnRHR (-1424.9 kcal/mol) and GPCR (-1451.8 kcal/mol). The results of the MD simulations revealed the significant stability of the spike protein with the GnRHR and GPCR up to 100 ns. The SARS-CoV-2 spike protein had strong binding interactions with the GPCRs and GnRHRs, which are highly expressed in the brain, endocrine organs, and olfactory neurons. This study paves the way towards understanding the complex mechanism of neuroendocrine involvement and peripheral organ involvement, may explain the changing symptoms in patients due to new variants, and may lead to the discovery of new drug targets for COVID-19. In vitro studies involving genetic engineering or gene knockdown of the GPCRs and GnRHRs are needed to further investigate the role of these receptors in COVID-19 pathogenesis.

Elhariri, M., R. Elhelw, S. Selim, M. Ibrahim, D. Hamza, and E. Hamza, "Virulence and Antibiotic Resistance Patterns of Extended-Spectrum Beta-Lactamase-Producing serovar Heidelberg Isolated from Broiler Chickens and Poultry Workers: A Potential Hazard.", Foodborne pathogens and disease, vol. 17, issue 6, pp. 373-381, 2020. Abstract

The current study investigated the emergence of multidrug-resistance (MDR), extended-spectrum beta-lactamase (ESBL)-producing serovar Heidelberg in broiler chickens and workers in poultry farms. A total of 33 . Heidelberg isolates were recovered; 24 from the broiler cloacal swabs and 9 from the farm workers. All the . Heidelberg isolates were tested for susceptibility to 11 antimicrobial agents and for the presence of resistance and virulence genes. MDR strains were found in 95.8% (23/24) and 88.8% (8/9) of the broiler and human isolates, respectively. Among the MDR strains, 66.6% of the broiler isolates and 55.5% of the human isolates were ESBL producing. The majority of broiler isolates showed resistance to ampicillin (100%) and ceftriaxone (91.6%), followed by ceftazidime and imipenem, (87.5%) and (75%). The resistance rate of the human isolates to those antibiotics were lower than the broiler isolates; ampicillin (88.8%), ceftriaxone (66.6%), ceftazidime (77.7%), and imipenem (66.6%). The resistance determinant genes found among the isolated strains was , , , , , , and . The most detected ESBL genes for broiler and human isolates were (63.7%) and (56.6%), followed by (48.5%), (39.4%), and (27.3%); whereas and were not detected. The finding of chromosomal and plasmid virulence genes revealed that the A (100%), , C, and (72.8%), C (66.7%), (63.6%), B (54.6%), and A and A (3.0%), while A and R were absent. An elevated rate of MDR Heidelberg in chickens is of potential great health risk. This signifies the role of the food of animal origin as a reservoir of MDR that can affect the human health.

Hamza, D., S. Dorgham, E. Ismael, S. I. A. El-Moez, M. Elhariri, R. Elhelw, and E. Hamza, "Emergence of β-lactamase- and carbapenemase- producing Enterobacteriaceae at integrated fish farms.", Antimicrobial resistance and infection control, vol. 9, issue 1, pp. 67, 2020. Abstract

BACKGROUND: Epidemiological studies suggested that determinants for antibiotic resistance have originated in aquaculture. Recently, the integrated agriculture-aquaculture system has been implemented, where fish are raised in ponds that receive agriculture drainage water. The present study aims to investigate the occurrence of β-lactamase and carbapenemase-producing Enterobacteriaceae in the integrated agriculture-aquaculture and the consequent public health implication.

METHODS: Samples were collected from fish, fishpond water inlets, tap water, outlet water, and workers at sites of integrated agriculture-aquacultures. Samples were also taken from inhabitants of the aquaculture surrounding areas. All samples were cultured on MacConkey agar, the Enterobacteriaceae isolates were tested for susceptibility to cephalosporins and carbapenems, and screened for bla, bla, bla, bla, bla, bla, bla, and bla. Strains having similar resistance phenotype and genotype were examined for the presence of Incompatible (Inc) plasmids.

RESULTS: A major proportion of the Enterobacteriaceae isolates were resistant to cephalosporins and carbapenems. Among the 66 isolates from fish, 34 were resistant to both cephalosporin and carbapenem groups, 26 to carbapenems alone, and 4 to cephalosporins alone. Of the 15 isolates from fishpond water inlets, 8 showed resistance to both groups, 1 to carbapenems alone, and 5 to cephalosporins alone. Out of the 33 isolates from tap water, 17 were resistant to both groups, and 16 to cephalosporins alone. Similarly, of the 16 outlet water isolates, 10 were resistant to both groups, and 6 to cephalosporins alone. Furthermore, of the 30 examined workers, 15 carried Enterobacteriaceae resistant strains, 10 to both groups, and 5 to cephalosporins alone. Similar strains were isolated from the inhabitants of the aquaculture surrounding areas. Irrespective of source of samples, strains resistant to all examined antibiotics, carried predominantly the carbapenemase gene bla either alone or with the β-lactamase genes (bla, bla, bla, and bla). The isolates from fish, water, and workers harboured a wide-range of multi-drug-resistance Inc. plasmids, which were similar among all isolates.

CONCLUSION: The present findings suggest transmission of the resistance genes among Enterobacteriaceae strains from different sources. This reiterates the need for control strategies that focus on humans, animals, water, and sewage systems to solve the antibiotic resistance problem.

Morsy, E. A., H. M. Salem, M. S. Khattab, D. A. Hamza, and M. M. Abuowarda, "Encephalitozoon cuniculi infection in farmed rabbits in Egypt.", Acta veterinaria Scandinavica, vol. 62, issue 1, pp. 11, 2020. Abstract

BACKGROUND: Encephalitozoon cuniculi is an important microsporidian parasite with zoonotic potential. The present study highlights the impact of encephalitozoonosis on rabbit health in Egypt. Three rabbit farms in Giza, with a total of 16,400 rabbits were investigated due to occurrence of rabbits displaying clinical signs consistent with encephalitozoonosis.

RESULTS: Clinical signs observed during a 4 months observation period in 2018 included vestibular disease, paresis, limb paralysis, cataracts, phacoclastic uveitis, frequent urination, marked decrease in body weight and in some pregnant females, also repeated abortions. The total morbidity rates in adult and young rabbits were 76.7% and 81.5%, respectively. The highest mortality rate was recorded in offspring (12.3%), followed by dams (5.6%), and the lowest recorded mortality rate was in males (0.04%). Post-mortem findings included enteritis, pale enlarged kidneys, congested leptomeninges, focal brain necrosis, and endometrial congestion. Histopathological examination revealed nonsuppurative meningoencephalitis and glial nodules with central necrosis in the brain, vacuolation and necrosis of renal tubular epithelium, and corneal ulceration and ruptured lens capsule with fragmentation of lenticular fibres. E. cuniculi were observed in the brain, retinal ganglion cells, kidneys, and liver. Transmission electron microscopy examination revealed the presence of different developmental stages of E. cuniculi in the brain and kidney. Presence of E. cuniculi was confirmed by conventional polymerase chain reaction using a universal 16S gene for Encephalitozoon spp. followed by sequencing and sequence analysis.

CONCLUSIONS: The presence of E. cuniculi in rabbits was confirmed at three farms in Egypt. Nervous signs and ocular lesions were the most predominant findings in these farms.

Hamza, D. A., R. M. Abd-Elsalam, S. M. Nader, M. Elhariri, R. Elhelw, and H. S. El-Mahallawy, "Pathways of Methicillin-Resistant in Animal Model: New Insights Regarding Public Health.", Infection and drug resistance, vol. 13, pp. 1593-1600, 2020. Abstract

Background: is considered one of the major threats regarding food safety worldwide. Methicillin-resistant (MRSA) strains in livestock, companion animals, and wild animals continue to be a potential risk to people working with them.

Aim: The current research aims to investigate the potential pathways of livestock-associated methicillin-resistant (LA-MRSA) strains in the body after oral infection using the experimental mouse model.

Methods: Seven groups of SPF male mice were purchased and housed. On day 1, six groups of mice were infected orally by the sterile gastric probe using 100 μL/mice of LA-MRSA bacterial suspension (1 × 10 colony-forming units (CFU)/mL). The remaining group was kept as negative controls. Over 15 days, these animals have been monitored. Fresh fecal samples were screened for LA-MRSA at day 0, day 7 and day 14 following oral administration of MRSA strains. All animals were sacrificed at day 15, and internal organs (liver, lung, kidney, and intestine) were harvested aseptically and divided into two sections. The first part was histopathologically investigated, while the other half has been tested for LA-MRSA re-isolation.

Result: The oral challenge of mice by MRSA strains showed that MRSA was re-isolated from feces and intestines of all inoculated mice groups and from internal organs (liver, lung, kidney and intestine) of most mice. Results were confirmed by the detection of the bacteria in gram-stained tissue sections and changes in H&E-stained histopathological tissue sections from these organs.

Conclusion: Data from the present study indicate the possible colonization of livestock-associated methicillin-resistant (LA-MRSA) in internal organs following oral infection and thus posing a risk for food-borne infection of MRSA. Infected animals could pass LA-MRSA through feces again, resulting in increased dispersion and environmental contamination.

Elshazly, M. O., S. S. A. El-Rahman, D. A. Hamza, and M. E. Ali, "Pathological and bacteriological studies on reproductive tract abnormalities of she-camels (), emphasizing on zoonotic importance.", Journal of advanced veterinary and animal research, vol. 7, issue 4, pp. 633-646, 2020. Abstract

Objective: Infertility caused by reproductive pathologies plays a significant role in animal breeding and could result in massive economic losses to livestock owners. Hence, this study was designed to allocate various pathological lesions in the female reproductive tract of she-camels () slaughtered in Egypt and isolate the causative agents associated with those pathologies.

Materials and Methods: A total of 500 genitalia of adult nonpregnant she-camels aged between 6 and 15 years old were collected from three slaughterhouses at the Giza Governorate, Egypt, from August 2017 to August 2019. The uterus, cervix, and vagina were examined pathologically and microbiologically.

Results: The uteri of 152 cases (30.4%), cervices of 24 cases (4.8%), and vaginae of 20 cases (4.2%) showed pathological abnormalities. The uterine inflammatory lesions were detected in 119 cases (23.8%), and the non-inflammatory lesions were detected in 58 cases (11.6%). Pathological changes of the cervix comprised 4.8%, whereas vaginal abnormalities represented 4%. The total microbial recovery rate was 28.4%, and the isolated organisms included , , , , and , in addition to . Trials to isolate and species were negative; however, virological examination revealed the isolation of bovine herpesvirus type-1 in two cases.

Conclusion: Inflammatory lesions were the most prevailing pathological lesions observed along the genital tract of she-camels, and was the most prevalent isolate. The microbiological burden from the genital discharge could be of zoonotic importance to the examiner and could be a contaminant to the environment and, consequently, human. In addition, attention should be paid toward the possibility of infected she-camels to transmit such infections to farm animals in contact.

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