Farghali, H. A., N. A. AbdElKader, H. O. AbuBakr, S. H. Aljuaydi, M. S. Khattab, R. Elhelw, and M. Elhariri, "Antimicrobial action of autologous platelet-rich plasma on MRSA-infected skin wounds in dogs.", Scientific reports, vol. 9, issue 1, pp. 12722, 2019. Abstractantimicrobial_action_of_autologous.pdf

Effective antimicrobial preparations, other than antibiotics, are important for the treatment of potentially fatal drug-resistant infections. Methicillin-resistant Staphylococcus aureus (MRSA) is one of the leading causes of hospital-acquired and post- operative infections. Fortunately, the antimicrobial properties of platelet-rich plasma (PRP) against various microorganisms enable its potential use as an alternative to conventional antibiotics. The present work was designed to evaluate the hypothesized antimicrobial activity of PRP against MRSA infected skin wounds. Six adult male dogs were divided equally into control and PRP groups. Unilateral circular full-thickness skin wounds were created then a MRSA suspension was injected locally. Treatment started at 1st week post infection with subcutaneous infiltration of autologous activated PRP every week in the PRP group and with topical application of clindamycin cream twice daily in the control group. PRP decreased wound size and significantly increased wound contractility and re-epithelization, as confirmed by histopathological and immunohistochemical findings. Also PRP treated group showed significant decrease in ROS and redox imbalance with over expression of the TNF-α and VEGFA genes that indicate angiogenesis and maximum antibacterial activity after three weeks. In conclusion, CaCl-activated PRP exhibited antimicrobial activity against MRSA infection, which improved the infected wound healing re-epithelization and granulation tissue formation.

Al-Amery, K., M. Elhariri, A. Elsayed, G. El-Moghazy, R. Elhelw, H. El-Mahallawy, M. El Hariri, and D. Hamza, "Vancomycin-resistant isolated from camel meat and slaughterhouse workers in Egypt.", Antimicrobial resistance and infection control, vol. 8, pp. 129, 2019. Abstractvancomycin-resistant_staphylococcus_aureus_isolated_from_camel_meat_and_slaughterhouse_workers_in_egypt.pdf

Background: The emergence of vancomycin-resistant (VRSA) represents a challenge for the treatment of staphylococcal infections in both human and animals worldwide. Although VRSA has been detected in several animal species worldwide, data on the bacterial prevalence in dromedary camels and workers in camel slaughterhouses are scarce.

Methods: We investigated meat samples from 200 dromedary camel carcasses from three different abattoirs that were being prepared to be sent to the markets. Twenty hand swabs were voluntarily collected from the workers in the same abattoirs. Isolation and identification of the bacterial specimens from the samples were performed using conventional cultural techniques and biochemical identification and were confirmed by PCR amplification of the gene. Antimicrobial susceptibility against nine antimicrobial agents commonly used in human and camels was tested using the disc diffusion method, and genetic analysis was performed by evaluating the gene in phenotypically oxacillin (OXA)- and cefoxitin (FOX)-resistant isolates. The resistance of to vancomycin (VAN) was tested by broth microdilution and confirmed by PCR targeting the and genes. The and genes were sequenced.

Result: was detected in both camel meat (29/200, 14.5%) and in abattoir workers (11/20, 55%). Of the collected samples, 27% (8/29, camel) and 54% (6/11, human) were identified as VRSA.All VRSA isolates carried both the and genes. Additionally, all VRSA isolates were also classified as methicillin-resistant (MRSA). The amplicons of the isolates from human and camel meat were homologous and clustered with a Chinese reference isolate sequence.

Conclusion: This study demonstrated that VRSA is present in camel abattoirs in Egypt. Zoonotic transmission between animals and human is probable and reflects both a public health threat and a food safety concern.

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, 2019. 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.

Elhariri, M., D. Hamza, R. Elhelw, and S. M. Dorgham, "Extended-spectrum beta-lactamase-producing Pseudomonas aeruginosa in camel in Egypt: potential human hazard.", Annals of clinical microbiology and antimicrobials, vol. 16, issue 1, pp. 21, 2017. Abstractextended-spectrum_beta-lactamase-producing_pseudomonas_aeruginosa_in_camel_in_egypt_potential_human_hazard..pdf

BACKGROUND: The rapid increase of extended-spectrum beta-lactamase (ESBL) producing bacteria are a potential health hazard. Development of antimicrobial resistance in animal pathogens has serious implications for human health, especially when such strains could be transmitted to human. In this study, the antimicrobial resistance due to ESBL producing Pseudomonas aeruginosa in the camel meat was investigated.

METHODS: In this study meat samples from 200 healthy camels at two major abattoirs in Egypt (Cairo and Giza) were collected. Following culture on cetrimide agar, suspected P. aeruginosa colonies were confirmed with a Vitek 2 system (bioMe´rieux). P. aeruginosa isolates were phenotypically identified as ESBL by double disk synergy test. Additionally antimicrobial susceptibility testing of ESBL producing P. aeruginosa isolates were done against 11 antimicrobial drugs and carried out by disk diffusion method. The ESBL genotypes were determined by polymerase chain reaction according to the presence of the bla , bla , bla , and bla .

RESULTS: Pseudomonas aeruginosa was isolated from 45 camel meat sample (22.5%). The total percentage of ESBL producing P. aeruginosa was 45% (21/45) from camel meat isolates. Antibiogram results revealed the highest resistance was for c, ceftriaxone and rifampicin followed by cefepime and aztreonam. The prevalence rates of β-lactamase genes were recorded (bla 28.5%, bla 38%, bla 33.3% and bla 23.8%).

CONCLUSIONS: This study illustrates the presence of high rates of ESBL-P. aeruginosa in camels that represents an increasing alarming for the risk of transmission to human and opens the door for current and future antibiotics therapy failure. Livestock associated ESBL-P. aeruginosa is a growing disaster, therefore, attention has to be fully given to livestock associated ESBL-bacteria which try to find its way to human beings.

Elhariri, M., D. Hamza, R. Elhelw, and E. Hamza, "Occurrence of cagA vacA s1a m1 i1 Helicobacter pylori in farm animals in Egypt and ability to survive in experimentally contaminated UHT milk.", Scientific reports, vol. 8, issue 1, pp. 14260, 2018. Abstractoccurrence_of_caga_vaca_s1a_m1_i1.pdf

Cases of human gastric cancer due to Helicobacter pylori have been reported worldwide and animals might act as a reservoir of infection in certain circumstances. The recent few decades showed a rapid decline in the incidence of gastric cancer, which was mainly due to the decrease in H. pylori infection. The aims of the present study were to determine the prevalence of H. pylori among livestock and investigate whether the animal isolates can be transmitted through contaminated milk causing gastric infection. Feces and milk samples were collected from apparently healthy cows, buffaloes, and sheep, and were examined by nested PCR and genotyping. The PCR positive samples were further subjected to bacterial culture followed by partial 16s sequencing of the isolates. Twenty-nine percent of the animals showed the presence of H. pylori, mainly the virulent cagAvacAs1a m1 i1 genotype, which is known to be associated with serious diseases in humans. The spiral viable culturable form (SVCF) of this strain was inoculated into UHT (ultra-high temperature) milk and remained viable for up to 10 days at 4 °C. Increasing period of storage and or temperature led to a decrease in the number of the SVCF and occurrence of the coccoid viable non-culturable form (CVNCF). The infectivity of the survived forms was determined by feeding healthy groups of laboratory mice with the contaminated UHT milk containing SVCF or CVNCF for 40 days. The gastric mucosa of the two mice groups showed similar levels of H. pylori load. This highlights that H. pylori can persist in contaminated milk by entering a non-culturable state, which can induce gastric infection.

Hamza, D., R. Elhelw, M. Elhariri, and E. Ragab, "Genotyping and antimicrobial resistance patterns of Helicobacter pylori in human and dogs associated with A2142G and A2143G point mutations in clarithromycin resistance.", Microbial pathogenesis, vol. 123, pp. 330-338, 2018. Abstractgenotyping_and_antimicrobial_resistance_patterns_of_helicobacter_pylori_in.pdf

BACKGROUND: Routes of transmission of Helicobacter pylori a class I carcinogen bacterium and the roles of animals have not been yet well determined. This study was carried out to investigate H. pylori phenotypically and genotypically in human and dogs to determine the antibiotic resistance patterns. As eradication therapy depends mainly on clarithromycin we evaluated 23S rRNA gene mutations associated with its resistance.

RESULTS: A total of 150 human stool samples and 60 canine gastric biopsies were examined by nested PCR for the presence of H. pylori, 60% and 76.6% were positive respectively. Only 20 (22.2%) and 41 (89.1%) isolates were successfully cultured from human and canine samples respectively. Genotyping revealed a total of cagAvacA combinations 76.6% (69/90) and 65.2% (30/46) in human and dogs, respectively. Allelic diversity in vacA gene was obviously observed, while cagAvacA combinations were 23.3% (21/90) and 34.7% (16/46) in human and dogs, respectively. The antimicrobial susceptibility patterns of human exhibited the highest levels of resistance against Clarithromycin (60%), Trimethoprim (55%), metronidazole (45%), amoxicillin (45%) and cefsulodin (60%) antibiotics and comparatively lower for spiramycin (10%) and tetracycline (15%). Dogs strains showed the highest levels of resistance against Clarithromycin (53.6%), metronidazole (51.2%) and erythromycin (43.9%) antibiotics, on the other hand, the percent of resistant canine strains were comparatively lower for spiramycin (9.7%). Single point mutation of A2143G was detected as 25% (3/12), 18.1% (4/22) in human and dogs respectively. Single point mutation of A2142G was detected as 16.6% (2/12), 13.6% (3/22) in human and dogs, respectively. While dual mutations of both A2142G and A2143G were detected as 50% (6/12), 40.9% (9/22) in human and dogs, respectively.

CONCLUSION: occurrence of elevated rates of A2142G and A2143G point mutations in clarithromycin resistant H. pylori isolates from human and dogs causing failure in treatment and eradication of the pathogen. The roles of animals need attention and further investigations.

Elhariri, M., R. Elhelw, D. Hamza, and H. S. El-Mahallawy, "Serologic evidence and risk factors for Helicobacter pylori infection in animals and humans.", Journal of infection in developing countries, vol. 11, issue 5, pp. 414-419, 2017. Abstractserologic_evidence_and_risk_factors_for_helicobacter_pylori_infection_in_animals_and_humans.pdf

INTRODUCTION: Helicobacter pylori (H. pylori) is one of the most common bacterial infections among humans worldwide. Although many records imply its interfamilial acquisition, the role of animals remains poorly understood. This study was undertaken to investigate the seroprevalence of H. pylori in animals and their human contacts in Cairo and Giza governorates, Egypt.

METHODOLOGY: Commercial enzyme-linked immunosorbent assay (ELISA) kits were used to detect IgG antibodies to H. pylori in dogs, cattle, and humans.

RESULTS: Seropositive dogs (35/94; 37.2%), cattle (24/80; 30%) and humans (40/90; 44.4%) were found. Seroprevalence in animals significantly varied in different areas of sample collection, but there was no association with sex or age. Human seropositivity rates were associated with increasing age; moreover, seropositive dog owners (51.7%; 15/29), had seropositive dogs. However, infection was not associated with subject's sex, occupation, or history of animal contact.

CONCLUSIONS: Our findings indicate H. pylori is widely distributed in cattle and dogs and their human contacts in Cairo and Giza, Egypt. Further studies to determine infection in other occupational groups are needed. This study provides baseline information on the seroprevalence of H. pylori, which may be required to begin prevention control programs in our area.

Hamza, D., S. M. Dorgham, M. Elhariri, R. Elhelw, and E. Ismael, "New Insight of Apparently Healthy Animals as a Potential Reservoir for : A Public Health Implication.", Journal of veterinary research, vol. 62, issue 4, pp. 457-462, 2018. Abstractnew_insight_of_apparently_healthy_animals_as_a_potential_reservoir_for_clostridium_perfringens_a_public_health_implication.pdf

Introduction: is commonly found in the gastrointestinal tract of animals and humans and continues to cause one of the most prevalent foodborne diseases in man.

Material and Methods: A total of 355 samples were examined for the occurrence of : rectal swabs from cattle, sheep, and goats, fresh stool samples from diarrhoea sufferers having been in contact with these animals, irrigation water and soil samples from the husbandry sites, and pre-harvesting fresh produce from farms irrigated with the sampled water. All samples were collected from Cairo and Giza governorates, Egypt. PCR analysis was carried out with positive isolates using the α-toxin gene. Sequence analysis of the gene of isolates was performed using the neighbour-joining approach. Bootstrap analysis was executed with 1,000 resamplings.

Results: 174 strains were isolated with a 49.01% prevalence. The highest prevalence of in apparently healthy animals was found in sheep (65.45%) followed by goats (58%), buffaloes (55%), and cattle (47.1%). Its prevalence in humans being in contact with these animals was 47.5%. The bacterium's isolation from the soil and irrigation water was achieved in 40% and 31.7% of samples, respectively, posing a risk, particularly when the water and soil contact food in the field, shown by the fresh produce isolation of 40%. A significant relationship between the prevalence of in animal and environmental samples was identified (P < 0.05). A significant relationship was identified neither between animal species and prevalence, nor between the environmental source and prevalence (P > 0.05). All isolates were positive for the α-toxin gene by PCR. The sequence analysis and the phylogenetic relationship of the α-toxin genes from different samples revealed that from faeces of apparently healthy cattle, buffaloes, sheep, and goats is a significant threat in places where it can contaminate the soil and water. In addition, the sequence of from humans suffering from diarrhoea was found in the same cluster with the sequence from cows, goats, and sheep.

Conclusion: The role of apparently healthy animals in transmitting to humans, either through being in direct or indirect contact water or soil in the cultivation of vegetables and fruits, was demonstrated.

Heidy Abo El-Yazeed, A. Nader, E. Fathy, M. Elhariri, R. Elhelw, and R. Soliman, "Molecular characterization of Clostridium perfringens isolated from broiler chickens in Egypt.", Bioscience research, vol. 15, issue 3, pp. 2312-2317, 2018.