Emergence of Virulent Extensively Drug-Resistant Vancomycin-Resistant Enterococci Among Diarrheic Pet Animals: A Possible Public Health Threat on the Move.

Background: Vancomycin-resistant enterococci (VRE) have become an increasing public health concern in the past few decades, being associated with serious multidrug-resistant (MDR) infections. This study was conducted to investigate the role of diarrheic pet animals as potential reservoirs for virulent extensively drug-resistant (XDR) VRE and their threat on human health. Materials and Methods: Rectal swabs were collected from 153 diarrheic pet animals (80 dogs and 73 cats). The collected swabs were cultured on CHROMagarTMVRE for the isolation of vancomycin-resistant Enterococcus faecalis and Enterococcus faecium, and then suspected colonies were identified as enterococci after Gram staining, conventional biochemical tests, and molecular techniques. VRE were basically identified using the disk diffusion method; however, molecular identification of vanA and vanB genes was carried out among confirmed VRE isolates. Moreover, three virulence genes (cytolysin A, cylA; enterococcal surface protein, esp; and hyaluronidase, hyl) were investigated in VRE isolates. Thereafter, VRE strains that harbored virulence genes were tested for antimicrobial susceptibility. Results: Eighteen out of 153 animals (11.8%) were positive for VRE, which were obtained from 15% and 8.2% of the examined dogs and cats, respectively. None of the obtained isolates carried the vanA gene, whereas the vanB gene was detected in E. faecalis (4/10) with a prevalence rate (40%). Of the obtained VRE isolates, five possessed esp and/or cylA, while all strains were negative for the hyl gene. Furthermore, four virulent VRE isolates exhibited an XDR pattern, and one isolate was MDR. Conclusion: Diarrheic pet animals could represent a potential zoonotic reservoir for virulent XDR vancomycin-resistant E. faecalis, which may have serious public health implications.

Occurrence of Major Human Extraintestinal Pathogenic Escherichia coli Sequence Types Among Diarrheic Pet Animals: A Potential Public Health Threat.

Background: Extraintestinal pathogenic Escherichia coli (ExPEC) has become a mounting public health concern. The present study was conducted to address the role of diarrheic pet animals as potential reservoirs for major human ExPEC sequence types (STs). Materials and Methods: Rectal swabs were collected from 145 diarrheic pet animals (75 dogs and 70 cats). Samples were processed for isolation and identification of E. coli by culture methods. Afterward, ExPEC isolates were identified on a molecular basis through detection of ExPEC phylogroups (B2 and D) coupled with carriage of two or more of the virulence genes associated with ExPEC (papAH, papC, sfa/focDE, afa/draBC, iutA, and kpsMT II). ExPEC STs 131, 73, 69, and 95 were identified among ExPEC isolates by quadruplex PCR and tested for their antimicrobial susceptibility. Eventually, two isolates underwent gene sequencing for the phylogenetic analysis. Results: Of 145 pet animals, 16 (11%) E. coli strains were identified as ExPEC, in which 15 (10.3%) isolates belonged to phylogroup B2 and 1 (0.69%) strain belonged to phylogroup D. The major human ExPEC STs were detected in 13 (9%) isolates, whereas the prevalence rates were 5.3% and 12.9% for dogs and cats, respectively. The isolation rates of ExPEC STs were 4.8%, 2.8%, 0.69%, and 0.69% for ST73, ST131, ST95, and ST69, respectively. Regarding the prevalence of virulence genes among ExPEC STs, the most prevalent ones were papC and sfa/focDE (92.3%), followed by papAH (76.9%), iutA (53.8%), afa/draBC (30.8%), and kpsMT II (30.8%). Moreover, 38.5% of the obtained human ExPEC STs were multidrug resistant. The phylogenetic analysis of two ExPEC ST73 gene sequences showed high genetic relatedness to those isolated from humans in different countries. Conclusions: The fecal carriage of major human ExPEC STs among diarrheic dogs and cats poses a potential zoonotic hazard with serious public health implications.

The Antibacterial Activity of Egyptian Wasp Chitosan-Based Nanoparticles against Important Antibiotic-Resistant Pathogens.

The current work discusses the production and characterization of new biodegradable nanoparticles for biomedical applications based on insect chitosan. Chitosan has numerous features due to the presence of primary amine groups in repeating units, such as antibacterial and anticancer activities. When polyanion tripolyphosphate is added to chitosan, it creates nanoparticles with higher antibacterial activity than the original chitosan. In this study, the ionic gelation technique was used to make wasp chitosan nanoparticles (WCSNPs) in which TEM and FTIR were used to investigate the physicochemical properties of the nanoparticles. In addition, the antibacterial activities of chitosan nanoparticles against extended-spectrum beta-lactamase (ESBL)- and carbapenemase-producing Klebsiella pneumoniae, Escherichia coli, and Pseudomonas aeruginosa were evaluated. The extracted wasp chitosan exhibited high solubility in acetic acid and met all standard criteria of all characterization testes for nanoparticles; the zeta potential indicated stable WCSNPs capable of binding to cellular membrane and increasing the cellular uptake. The produced WCSNPs showed growth inhibition activity against all tested strains, and the bacterial count was lower than the initial count. The inhibition percent of WCSNPs showed that the lowest concentration of WCSNPs was found to be effective against tested strains. WCSNPs' antibacterial activity implies that they could be used as novel, highly effective antibacterial agents in a variety of biological applications requiring antibacterial characteristics.

The Public Health Burden of Virulent Extended-Spectrum ß-Lactamase-Producing Klebsiella pneumoniae Strains Isolated from Diseased Horses.

Background:Klebsiella pneumoniae has been associated with both nosocomial and community-acquired infections with mounting public health concern throughout the world. The purpose of this study was to investigate the burden of virulent extended-spectrum ß-lactamase (ESBL)-producing K. pneumoniae among diarrheic horses or those with respiratory illness to underscore the public health implication of such strains. Materials and Methods: Rectal and nasal swabs were gathered from 100 diseased horses (50 diarrheic and 50 with respiratory illness). The collected swabs were processed for isolation of ESBL-producing K. pneumoniae using a selective medium followed by phenotypic and molecular identification of the isolates. All ESBL-producing K. pneumoniae strains were investigated for six virulence genes (type 3 fimbrial adhesin [mrkD], enterobactin [entB], regulator of mucoid phenotype A [rmpA], Klebsiella ferric iron uptake [kfu], mucoviscosity-associated gene A [magA], and type 2 capsular polysaccharide [K2]). Results: Of the 100 examined animals, ESBL-producing K. pneumoniae was recovered from 13 (13%), with isolation rates in horses suffering from diarrhea and respiratory illness being 20% and 6%, respectively. Among the obtained isolates, bla TEM and bla SHV were found in all strains (100%) followed by bla CTX-M in 92.3%, while none of the isolates had bla OXA. In addition, 13 ESBL-producing K. pneumoniae strains exhibited a multidrug resistance (MDR) pattern. Regarding the occurrence of virulence genes among the isolates, mrkD (100%) and entB (100%) were the most predominant virulence genes followed by rmpA (76.9%) and kfu (46.2%). On the contrary, magA and K2 were negative in all ESBL-producing strains. Furthermore, this work provides four K. pneumoniae mrkD partial sequences that displayed high genetic relatedness with those obtained from human to clarify the public health burden of such isolates. Conclusion: The occurrence of virulent ESBL-producing K. pneumoniae among diseased horses highlights the potential role of this animal in the epidemiology of such virulent and antimicrobial-resistant strains, which may have great public health threat.

Molecular Detection of Toxigenic Clostridioides difficile among Diarrheic Dogs and Cats: A Mounting Public Health Concern.

Nowadays, pet animals are known to be asymptomatic carriers of Clostridioidesdifficile. This study was conducted to investigate the burden of toxigenic C. difficile among diarrheic dogs and cats using direct PCR on fecal samples to reveal better insights about the epidemiology of such toxigenic strains referring to its public health significance. For this purpose, fecal samples were obtained from 58 dogs and 42 cats experiencing diarrhea. Following DNA extraction, the extracted DNA was examined for the occurrence of C. difficile as well as toxigenic strains through the detection of C. difficile 16S rRNA and toxin encoding genes (tcdA, tcdB, cdtA and cdtB) using PCR. Moreover, partial DNA sequencing of toxigenic strains retrieved from dog and cat was carried out. Of 100 examined diarrheic animals, 90 (90%) were C. difficile positive, including 93.1% and 85.7% of dogs and cats, respectively. In addition, toxigenic strains were detected in 13 animals, giving an overall prevalence 13% with the following prevalence rates among dogs and cats 12.1% and 14.3%, respectively. Furthermore, the phylogenetic analysis of the obtained sequence revealed high genetic relatedness of tcdA sequence obtained from a cat to strains of human diarrheic cases to point out the public health threat of such sequence. In conclusion, the direct detection of toxigenic C. difficile using PCR among dogs and cats highlights the potential role of household pets as a source for such strains to human contacts.

Parturient Cat As a Potential Reservoir for Coxiella burnetii: A Hidden Threat to Pet Owners.

Background: Q fever is a worldwide zoonosis caused by Coxiella burnetii. This study was carried out to investigate the occurrence of C. burnetii among apparently healthy pregnant, parturient, and postparturient dogs and cats to highlight their role in the transmission of such disease to humans. Materials and Methods: A total of 88 apparently healthy pet animals (48 dogs and 40 cats) were enrolled in this study, vaginal swabs were obtained from pregnant and postparturient animals while birth fluids were collected from parturient ones. All samples were subjected to DNA extraction followed by nested PCR for molecular detection of C. burnetii. Results: Out of 40 cats, 3 were positive for C. burnetii with an overall prevalence of 7.5%, all positive samples were birth fluids of parturient queens with a prevalence of 15.8% (3/19) while all pregnant and postparturient animals were negative. In contrast, none of 48 dogs yielded positive result. Moreover, the phylogenetic analysis and sequence identity matrix of the obtained sequence from a parturient cat showed high genetic relatedness to strains derived from human cases rather than those of ruminants to indicate the public health burden of such strain. Conclusion: This study underscores the occurrence of C. burnetii among parturient cats to point out the possible zoonotic transmission to human contacts.

The Nasal Carriage of Coagulase-Negative Staphylococci Among Animals and Its Public Health Implication.

Background: The research scope toward nasal colonization of coagulase-negative staphylococci (CoNS) in animals is largely ignored for many years. Therefore, this study was conducted to investigate the nasal carriage of CoNS among different animals and its public health implication. Materials and Methods: Nasal swabs were gathered from 152 animals (36 cats, 31 dogs, 29 sheep, 32 goats, and 24 cattle). These samples were subjected for isolation and identification of CoNS by conventional bacteriological methods, then molecular confirmation was carried out using Staphylococcus genus-specific 16S rRNA PCR. All CoNS isolates were screened for the presence of antibiotic resistance (mecA and blaZ) and virulence (lukS/F-PV and tsst-1) genes. Moreover, strains carrying resistance and/or virulence genes were identified to species level by 16S rRNA gene sequencing approach. Results: CoNS were identified in 14.5% (22/152) of the examined animals, whereas the prevalence rates among different animals were 27.8%, 3.2%, 8.3%, 10.3%, and 18.8% for cats, dogs, cattle, sheep, and goats, respectively. Of all isolates, two strains (Staphylococcus epidermidis and Staphylococcus warneri) harbored mecA gene, which carried on staphylococcal cassette chromosome mec type I in S. epidermidis and type V in S. warneri, while blaZ gene has been found in one strain (Staphylococcus felis). Importantly, two isolates (S. epidermidis and S. felis) had tsst-1 gene but all of CoNS isolates were negative for Panton-Valentine leukocidin gene. The phylogenetic analysis of the obtained sequences of CoNS of the current study revealed high similarity to those of serious human clinical cases to underscore the public health significance of such isolates. Conclusion: The nasal carriage of antibiotic-resistant and toxigenic CoNS among different animals highlights the potential zoonotic link with great public health implication.

Emergence of penicillin-macrolide-resistant Streptococcus pyogenes among pet animals: An ongoing public health threat.

Macrolide-resistant Streptococcus pyogenes is an emerging problem with a great public health concern throughout the world. The current study was carried out in order to investigate the possible role of pet animals in the epidemiology of such pathogen. For this purpose, nasal or oral swabs were collected from 115 pets (40 dogs and 75 cats) with respiratory illness. The collected swabs were cultured for isolation and identification of S. pyogenes. Macrolide-resistant S. pyogenes strains were initially identified after antibiotic susceptibility testing of the all obtained S. pyogenes isolates, then the phenotypic and molecular identification were done using the double-disk test and the detection of macrolide resistance genes, respectively. Of the 115 examined pet animals, S. pyogenes was recovered from 11 (9.6 %), from which, the isolation rates among dogs and cats were 15 % and 6.7 %, respectively. Macrolide-resistant S. pyogenes was isolated from dogs and cats in the following rates 10 % and 5.3 %, respectively. All macrolide-resistant S. pyogenes strains were assigned to cMLS resistance phenotype while all of them carried ermB gene only, except one strain from a cat possessed both ermB and ermTR genes. The phylogenetic analysis of 4 ermB gene sequences showed high genetic relatedness with those carried by bacteria isolated from human cases to underline the public health impact of such strains. Seriously, all macrolide-resistant S. pyogenes strains were resistant to penicillin. The emergence of penicillin-macrolide resistant S. pyogenes among pet animals underscores not only an emerging veterinary pathogen, but also an ongoing public health threat.

Occurrence of multidrug-resistant methicillin-resistant Staphylococcus aureus among healthy farm animals: a public health concern.

Methicillin-resistant Staphylococcus aureus (MRSA) is an emerging pathogen causing serious public health threats. This study was conducted to investigate the occurrence of multidrug-resistant MRSA among apparently healthy farm animals to shed the light on the potential role of these animals as a reservoir for such pathogen. For this purpose, 195 nasal swabs from apparently healthy farm animals (52 sheep, 51 goats, 47 cattle and 45 buffalo) were screened for multidrug-resistant MRSA. MRSA was isolated using a selective chromogenic medium and identified by colonial characters, Gram's stain films, conventional biochemical tests, coagulase test, resistance to cefoxitin and amplification of nuc and mecA genes. The antimicrobial susceptibility testing profile was performed by disk diffusion method to identify multidrug-resistant MRSA. Of 195 samples, 7 yielded MRSA with an overall prevalence 3.6%, whereas the prevalence rates were 3.8%, 3.9%, 4.3% and 2.2% for sheep, goats, cattle and buffalo, respectively. All MRSA isolates were multidrug-resistant strains. The phylogenetic analysis of 2 mecA gene sequences from the obtained isolates revealed that both sequences were clustered in the same clade with those derived from human clinical cases from different countries to highlight the public health burden of such strains. The distribution of multidrug-resistant MRSA among all examined farm animal species being apparently healthy points out that farm animals could represent a potential reservoir for multidrug-resistant MRSA with public health implications.

Camel as a transboundary vector for emerging exotic Salmonella serovars.

The current study was conducted to shed light on the role of imported camels as a transboundary vector for emerging exotic Salmonella serovars. Fecal samples were collected from 206 camels directly after slaughtering including 25 local camels and 181 imported ones as well as stool specimens were obtained from 50 slaughterhouse workers at the same abattoir. The obtained samples were cultured while Salmonella serovars were identified through Gram's stain films, biochemical tests and serotyping with antisera kit. Moreover, the obtained Salmonella serovars were examined by PCR for the presence of invA and stn genes. The overall prevalence of Salmonella serovars among the examined camels was 8.3%. Stn gene was detected in the vast majority of exotic strains (11/14) 78.6% including emerging serovars such as Salmonella Saintpaul, S. Chester, S. Typhimurium whereas only one isolate from local camels carried stn gene (1/3) 33.3%. On the other hand, none of the examined humans yielded positive result. Our findings highlight the potential role of imported camels as a transboundary vector for exotic emerging Salomenella serovars.

Molecular Detection of Francisella spp. Among Ticks Attached to Camels in Egypt.

This study was conducted to investigate the possible role of camels and attached ticks in the epidemiology of Francisella spp. including Francisella tularensis. For this purpose, a total of 319 ticks (248 Hyalomma dromedarii and 71 Amblyomma spp.) as well as 100 blood and 50 fecal samples collected from camels were screened for the presence of Francisella spp. by PCR through amplification of Francisella 16S rRNA gene. Positive samples were then tested for F. tularensis by PCR. In addition, serum samples from 75 camel abattoir workers were examined for the presence of IgG antibodies against F. tularensis using enzyme-linked immunosorbent assay (ELISA). Of the examined ticks, 15 were positive for Francisella spp. with prevalence of 4.7%, all positive results were recorded in Hyalomma dromedarii (6%). Neither blood nor fecal samples from camels yielded Francisella spp. even camels which carried Francisella spp. positive ticks. Moreover, F. tularensis could not be detected among Francisella-positive ticks. Phylogenetic analysis of some Francisella 16S rRNA gene sequences obtained in this study points out that these sequences are closely related to Francisella-like endosymbionts. In contrast, seroprevalence of F. tularensis antibodies among examined abattoir workers was 9.3% with significantly high prevalence among workers frequently exposed to tick bites (20.7%) rather than occasionally exposed workers (2.2%). In conclusion, however, F. tularensis could not be detected in this study; the high seroprevalence among camel abattoir workers especially those frequently exposed to tick bites underlines the possible role of ticks attached to camels in transmission of tularemia to humans.