Isolation and Antibiogram Profiles of Staphylococcus aureus Isolates from Cow milk and Dog samples

Staphylococcus aureus (S. aureus) is a commensal bacterium associated with serious infections in men and animals. Recently, multidrug-resistant (MDR) strains of S. aureus especially the so-called methicillin-resistant S. aureus (MRSA), represent a serious challenge that hinders the control of infections in man and animals. This study aimed to monitor milk samples from mastitic cows and vaginal and nasal swabs from dogs for the incidence of S. aureus. In addition, the isolates' antibiogram profiles were assessed to determine the extent of MDR and MRSA existence among the recovered isolates. Out of 260 samples, 29 (11%) S. aureus isolates were recovered with the highest incidence in milk samples (15/90, 17%), followed by vaginal swabs (8/90, 9%) and nasal swabs (6/80, 7%). Identification of the isolates was confirmed by PCR amplification of 16S rRNA gene sequence. Twenty S. aureus isolates were tested against seven antibacterial agents. Surprisingly, all the twenty isolates were MRSA and three bitch vaginal isolates were MDR. The findings of this study call for more research and cooperation between authors interested in assessing the MRSA and MRD bacterial incidence in both medical and veterinary fields. The cooperation will augment the challenge of disseminating MRSA and MDR staphylococci from animals to humans and vice versa

Incidence, Bacterial causes and Antibiotic Resistance Patterns of Urinary Tract Infection in Pet Animals

T The primary goal of the study was to determine the prevalence and various bacterial risks of lower urinary tract infections (UTI) in diseased and seemingly healthy pet animals with and without urine retention whether they were catheterized or not. The bacterial isolates were in vitro tested for their antibiotic resistance and antibiotic resistance genes were investigated. Between October 2020 and January 2022, 128 urine samples were randomly collected from pets recruited to veterinary hospitals and clinics in Cairo and Giza. Samples were cultivated for bacteriological isolation. Escherichia coli (E. coli), Staphylococcus aureus (S. aureus) and Klebsiella spp. were found to be the most common bacterial causes of urinary tract infections in pets, with prevalence rates of 32.9%, 28%, and 19.5%, respectively followed by Proteus mirabilis (P. mirabilis) and Pseudomonas aeruginosa (P. aeruginosa) with incidences of 18.2% and 1.2%, respectively. Based on bacterial types and their virulence genes, antibiotic resistance and multi-drug resistance (MDR) behaviour varied. Epidemiology, diagnosis, and control of the urinary tract infection would benefit from the identification and characterization of isolated bacterial species

Cinnamon oil downregulates virulence genes of poultry respiratory bacterial agents and revealed significant bacterial inhibition: An in vitro perspective

Background and Aim: Respiratory bacterial agents represent one of the most harmful factors that ordinarily threaten the poultry industry and usually lead to great economic losses. Meanwhile, there is a global demand to avoid the highly emerging antibiotic resistance and antibiotic residues in edible meat. Whereas, the use of alternatives became of great priority, especially for those substances extracted from natural plant origin. The study aimed to evaluate the antibacterial effect of cinnamon oil as a herbal extract on different respiratory bacterial agents. Materials and Methods: One hundred and fifty biological samples were collected through targeted surveillance for respiratory diseased poultry farms representing three governorates, from which bacterial isolation and identification, DNA sequencing of representative strains were performed. Furtherly, phenotypic and genotypic evaluation of the antibacterial effect of cinnamon oil was performed by minimum inhibitory concentration, agar disk diffusion, and virulence genes expression real-time polymerase chain reaction. Results: Cinnamon oil gave rise to acceptable degrees of virulence genes downregulation of 0.15, 0.19, 0.37, 0.41, 0.77, and 0.85 for Staphylococcus aureus sed gene, Escherichia coli stx1 gene, Avibacterium paragallinarum HPG-2 gene, Pasteurella multocida ptfA gene, Mycoplasma gallisepticum Mgc2 gene, and Ornithobacterium rhinotracheale adk gene, respectively. Phenotypically, using agar disk diffusion assay and broth microdilution susceptibility, cinnamon oil showed also tolerable results as it stopped the growth of S. aureus, E. coli, P. multocida, and A. paragallinarum with varying zones of inhibition. Conclusion: The encountered results declared the successful in vitro effect of cinnamon oil that recommends its application for living birds for future use as a safe antibacterial in the poultry industry

Pasteurellaceae members with similar morphological patterns associated with respiratory manifestations in ducks

Aim: A total of 112 freshly dead ducks aged from 2 to 20 weeks old with a history of respiratory manifestations were investigated for the implication of Pasteurellaceae family members.. Materials and Methods: Isolation and identification to the family level were conducted by conventional bacteriological methods, including microscopic examination and biochemical characterization. Identification to the species level was conducted by polymerase chain reaction (PCR) and analytical profile index (API) 20E kits. Results: Conventional bacteriological isolation and biochemical characterization revealed the infection of 16/112 examined birds with a prevalence rate of 14.3%. PCR confirmed the detection of Pasteurellaceae family conserved genes RpoB and Bootz in 16/16 (100%) isolates. PCR was also used for genus and species identification of the isolated Pasteurellaceae members; the results revealed that 5/16 (31.3%) of isolates were Gallibacterium anatis and 2/16 of isolates (12.5%) were Pasteurella multocida. Riemerella anatipestifer, Mannheimia haemolytica, and Avibacterium paragallinarum were not detected by PCR. Biotyping by API 20E successfully identified 5/16 (31.3%) isolates that could not be typed by PCR and confirmed their belonging to Pasteurella pneumotropica. Neither the available PCR primer sets nor API 20E succeeded for species identification of 4/16 (25%) isolates. Antibiotic susceptibility profiling of isolates revealed that 16/16 (100%) of isolates demonstrated multidrug resistance (MDR) phenotypes. Moreover, 16/16 (100%) of isolates demonstrated a phenotypic resistance pattern to neomycin. Conclusion: Combined genotypic, phenotypic, biotyping, and virulence characterizations are required for laboratory identification of pathogenic Pasteurellaceae. Moreover, P. multocida was not the prevailed member implicated in respiratory problems in ducks as P. pneumotropica, G. anatis, and unidentified strains were involved with higher prevalence. Chloramphenicol and ampicillin demonstrated the highest in vivo effects on the studied Pasteurellaceae. Furthermore, the prevalence of multidrug-resistant isolates signified the demand to implement targeted surveillance in the ducks' production sector, and MDR survey in poultry sectors in Egypt to apply effective control measures

Phenotypic and Genotypic Characteristics of Antimicrobial Resistance of Gram-negative Bacteria isolated From Pet Animal

Most animal feeds are set from protein-rich raw materials. These protein constituents may possess various hazards, particularly highly drug-resistant pathogens, causing a bad impact not only on the pet's health, but also on their owners. In the current study, a total of 2100 pet food and 100 pets’ fecal swabs were collected and bacteriologically examined from 2017 to 2020. It was revealed that the percentage of Gram-negative bacteria isolated from pet food and fecal swabs was 49% and 56% respectively. E. coli, Proteus sp., and K. pneumoniae were the most isolated bacteria in percentages of 12.4%, 8.4%, and 4.9% respectively from Pet food and 25%, 7%, 12% respectively from pet fecal swabs. In addition, Enterobacter cloacae, P. aeruginosa, Aeromonas hydrophila, Citrobacter sp., P. fluorecens, and Y. enterocolitica were isolated from pet food in order to 3.8%, 3.5%, 3.2%, 2.6%, 2.6% and 2.1% respectively. Salmonella sp. isolated from pet food was 0.6% while it was 5% from pet fecal swabs. The most predominant salmonella serotype isolated from pet food and pet fecal swabs was S. Typhimurium. Furthermore, S. Virchow, S. Anatum, S. Kentucky, S. Kedougou and S. Infantis were isolated serotypes from Pet food in percentages of 15.7%, 23.1%, 15.4%, 7.7%, and 7.7% respectively. While S. Nitra, S. Ibargi, S. Enteritidis and S. Boecker were isolated from pet fecal swabs at a percentage of 20% for each. On the other hand, O158 was the most predominant E. coli serogroup isolated from pet food and pet fecal swabs in percentages of 30.4% and 30.8% respectively followed by O157 in percentages of 21.7% and 26.9% respectively. O26 was isolated from pet food and pet fecal swabs in percentages of 13% and 7.7% for each. O119 was isolated from pet food and pet fecal swabs in percentages of 4.3% and 3.8% respectively. O86, O27, O44, O55, and O78 were isolated from pet food in the percentage of 4.3%, 8.7%, 4.3%, 4.3%, and 8.7%respectively. While O114, O111, and O125 were isolated serotypes from pet fecal swabs in percentages of 15.4%, 3.8%, and 11.5% respectively. This study revealed that the antimicrobial sensitivity test of 80% of Salmonellae were resistant to Cefotaxime and Colistin sulphate while 50%, 30, and 20% of isolates were resistant to Gentamicin, Tetracycline, and Cefepime respectively, while 40% of Salmonellae were resistant to Chloramphenicol, Enrofloxacin, and Amoxicillin-clavulanate. Also 60% of Salmonellae showed resistance to Trimethoprim sulfamethoxazole and Ciprofloxacin. Detection of Extended-spectrum ß-lactamase resistance genes (blaTEM, blaSHV, and blaCTX-M) in Pets using Polymerase chain reaction (PCR) showed the presence of blaTEM and blaSHV genes in all tested isolates in 12 samples out of 12 (100%) and has shown that the ratio of blaCTX-M is 5 out of 12samples (41.6 %)

The occurrence of disinfectant and antibiotic-resistant genes in Escherichia coli isolated from chickens in Egypt

Aim: This work aimed to determine the occurrence of antibiotic and disinfectant resistance genes in Escherichia coli isolated from chickens in Egypt. Materials and Methods: Organs (liver, lung, heart, yolk sac, and bone marrow) of 1500 chicken samples were collected from diseased chickens suffered from colibacillosis with PM findings as CRD, diarrhea and omphalitis from different governorates of Egypt as: Giza, EL-Bahira, Fayoum, El-Dakahlia, El-Ismalia, and El-Sharkia during 2015-2016. These samples were labeled and transported immediately on ice to the Reference laboratory for quality control on poultry production (RLQP). The samples were cultured onto MacConkey agar and Eosin Methylene Blue Agar. Isolation and identification of the E. coli were performed based on morphology, cultural, staining, and biochemical properties. Antimicrobial resistance test was carried out using disk diffusion method. The PCR employing tetA, qacED1 and qacA/B were carried out for detection of these genes in isolated E.coli. Results: The prevalence of E. coli in chicken was 34%. Predominant serotypes of E. coli which serologically identified were O128, O111, O44, O158, and O2. Antibiotic susceptibility test of E. coli revealed that 100% of isolates were resistant to ampicillin, erythromycin, and sulfamethoxazole-trimethoprim, while 73.53% and 38.23% of them were sensitive for colistin sulfate and levofloxacin, respectively. Antibiotic resistance genes as tetA gene were tested for isolated E. coli and detected by incidence rate of 91.18%. qac resistance genes resembling as qacED1 and qacA/B genes were detected in isolated E. coli 70.6% and 14.7%, respectively. Conclusion: E. coli isolated from chickens in Egypt was carried qac and antibiotic-resistant genes that affect the poultry industry

Molecular detection of multidrug-resistant Pseudomonas aeruginosa of different avian sources with pathogenicity testing and in vitro evaluation of antibacterial efficacy of silver nanoparticles against multidrug-resistant P. aeruginosa

ABSTRACT: Pseudomonas aeruginosa (P. aeruginosa) is a serious zoonotic pathogen threaten the poultry industry causing severe economic losses therefor, this study aimed to isolation, phenotypic, molecular identification of P. aeruginosa from different avian sources (chickens, turkey, pigeons, table eggs, and dead in shell chicken embryos), from different Egyptian governorates (Giza, Qalubia, Beheira, El-Minya, and Al-Sharqia) with applying of antibiotic sensitivity test on all P. aeruginosa isolates. Highly resistant isolates (n = 49) were subjected to molecular identification of P. aeruginosa with detection of resistant genes including carbapenemase-encoding genes blaKPC, blaOXA-48, and blaNDM. On the base of molecular results, a highly resistant P. aeruginosa strain was tested for its pathogenicity on day old specific pathogen free (SPF) chicks. Also, in vitro experiment was adopted to evaluate the efficacy of silver nanoparticles (Ag-NPs) against highly antibiotic-resistant P. aeruginosa strains. The overall isolation percentage was from all examined samples were 36.2% (571/1,576) representing 45.2% (532/1,176) from different birds' tissues and 39/400 (9.7%) from total egg samples. Some of isolated strains showed multidrug resistance (MDR) against kanamycin, amoxicillin, amoxicillin-clavulanic acid, neomycin, chloramphenicol, vancomycin, cefotaxime clavulanic acid, lincomycin-spectinomycin, co-trimoxazole, cefoxitin, gentamycin, and doxycycline. These MDR strains were also molecularly positive for ESBL and carbapenemase-encoding genes. MDR strain showed high pathogenicity with histopathological alterations in different organs in challenged birds. Main histopathological lesions were necrosis of hepatocytes, renal tubular epithelium, and heart muscle bundles. The MDR strain showed in vitro sensitivity to Ag-NPs. In conclusion, MDR P. aeruginosa is a serious pathogen causing high morbidity, mortality, and pathological tissue alterations. Ag NPs revealed a promising in vitro antimicrobial sensitivity against MDR P. aeruginosa and further in vivo studies were recommended

Pathogens Removal in a Sustainable and Economic High-Rate Algal Pond Wastewater Treatment System

This study evaluates the efficiency of a sustainable technology represented in an integrated pilot-scale system, which includes a facultative pond (FP), a high-rate algal pond (HRAP), and a rock filter (RF) for wastewater treatment to produce water that complies with the Egyptian standards for treated wastewater reuse. Still, limited data are available on pathogen removal through HRAP systems. Thus, in this study, the performance of the integrated system was investigated for the removal of Escherichia coli (E. coli), coliform bacteria, eukaryotic pathogens (Cryptosporidium spp., Giardia intestinalis, and helminth ova), somatic coliphages (SOMCPH), and human adenovirus (HAdV). Furthermore, physicochemical parameters were determined in order to evaluate the performance of the integrated system. The principal component analysis and non-metric multidimensional scaling analysis showed a strong significant effect of the integrated system on changing the physicochemical and microbial parameters from inlet to outlet. The mean log10 removal values for total coliform, fecal coliform, and E. coli were 5.67, 5.62, and 5.69, respectively, while 0.88 log10 and 1.65 log10 reductions were observed for HAdV and SOMCPH, respectively. The mean removal of Cryptosporidium spp. and Giardia intestinalis was 0.52 and 2.42 log10, respectively. The integrated system achieved 100% removal of helminth ova. The results demonstrated that the system was able to improve the chemical and microbial characteristics of the outlet to acceptable levels for non-food crops irrigation. Such findings together with low operation and construction costs of HRAPs should facilitate wider implementation of these nature-based systems in remote and rural communities. Overall, this study provides a novel insight into the performance of such systems to eliminate multiple microbial pathogens from wastewater

Zoonotic risk and public health hazards of companion animals in the transmission of Helicobacter species

Objective: Helicobacteriosis is worldwide infection caused by Helicobacter species that affects both humans and animals. The current work correlated the zoonotic and public health repertoire of Helicobacter species in companion animals (dogs and cats). Methods: Samples were collected from apparently healthy dogs (70), cats (65), and 70 human patients who had been in contact with these animals in the Cairo and Giza governorates. The samples included serum, feces, and stool samples and biopsies of gastric fundus fragments (~5 mm). All samples were examined by culture, biochemical analysis, serology, and molecular identification. Results: Helicobacter species were detected at a rate of 43.4% by PCR. H. heilmannii was more predominant, with a rate of 16%, whereas H. pylori was detected at 6%. H. pylori and H. heilmannii were isolated from both human and companion samples, whereas all samples were negative for H. felis. Conclusion: Dogs and cats were reservoirs and played a major source in human helicobacters infection