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.

Retail chicken giblets contaminated with extended-spectrum cephalosporin- and carbapenem-resistant Salmonella enterica carrying blaCMY-2.

Background and Aim: Chickens are considered as the main source of Salmonella, with infection potentially spreading to the public through outlets. The study aimed to investigate poultry shops for Salmonella enterica resistant to extended-spectrum cephalosporins-resistant (ESCR) and carbapenems-resistant (CR). Materials and Methods: Samples were collected from chicken giblets, water tanks, and workers at retail shops. Salmonella was isolated and serotyped; the presence of invA, stn, ompA, and ompF was determined using polymerase chain reaction (PCR). The isolates were tested for ESCR and CR by a disk-diffusion test; a confirmatory extended-spectrum ß-lactamase (ESBL) test was performed by combinational disk-diffusion test with clavulanic acid. The resistant isolates were screened for ESBL (blaTEM, blaSHV, blaCTX-M, and blaOXA-1), AmpC blaCMY-2, and carbapenemase (blaKPC, blaNDM, and blaOXA-48) genes using PCR. Results: S. enterica was isolated from chicken giblets (13/129) and the 13 isolates were ESCR. Based on the confirmatory ESBL test and CR, the 13 isolates were classified into the following resistance phenotypes: ESBL-producing and CR (n=4), ESBL-producing (n=1), non-ESBL-producing and CR (n=6), and non-ESBL-producing (n=2). All the five isolates with ESBL-producing phenotype carried predominantly blaTEM, blaSHV, and blaCMY-2. Regardless of being phenotypically CR, none of these isolates carried any of the tested carbapenemase genes. Surprisingly, the isolates with non-ESBL phenotype were found to carry blaTEM, blaSHV, and blaCMY-2. The blaKPC was present mainly in the isolates with non-ESBL and CR phenotypes. Interestingly, two isolates of the non-ESBL and CR phenotype showed resistance to cefepime, the fourth generation cephalosporins. Salmonella was also recovered from the water tanks (2/7) and the workers (2/16). The four isolates were ESCR and showed a non-ESBL-producing and CR phenotype; they harbored blaTEM, blaSHV, blaOXA-1, and blaKPC. The blaCMY-2 was found in one isolate from water and one from humans. All Salmonella isolates carried invA, stn, ompA, and ompF. Conclusion: Virulent ESCR S. enterica were identified in retail shops. The isolates carried blaCMY-2 and ESBL-genes, with a high proportion showing CR. Transmission of such strains to humans through food leads us to recommend regular inspection of retail outlets for antibiotic-resistant bacteria.

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.

Migratory birds as a potential overseas transmitter of Shiga toxin-producing Escherichia coli.

Shiga toxin-producing Escherichia coli (STEC) has a great public health importance. This study was conducted to investigate the potential role of migratory birds in the transmission of STEC. For this purpose, cloacal swabs were collected from 349 migratory birds (209 ducks and 140 quails) from Damietta governorate, Egypt. The collected swabs were cultured for isolation of STEC using the STEC CHROMagar. STEC isolates were identified based on colonial characteristics, Gram's stain, conventional biochemical tests and molecular detection of stx1, stx2 and eae genes. Positive isolates were serotyped and examined for their antibiotic susceptibility pattern. Furthermore, gene sequencing was performed for genes stx1and stx2. Of the examined birds, two STEC isolates were a obtained with an overall occurrence rate 0.57% (2/349), one isolate carried stx2 gene from a migratory quail 0.71% (1/140), and another isolate from a migratory duck carried stx1 gene 0.48% (1/209), whereas both isolates were negative for eae gene. Moreover, the duck isolate was serotyped O86, while the quail isolate was serotyped O125; both isolates were multidrug resistant. The phylogenetic analysis of the obtained stx1 and stx2 genes revealed high genetic relatedness to those isolated from human cases in the countries where such birds either lived or were in their migratory pathway. In conclusion, this study highlights the potential role of migratory birds in transmitting multidrug-resistant STEC across their migratory pathway.

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.

Extended-spectrum ß-lactamase-producing Salmonella serovars among healthy and diseased chickens and their public health implication.

OBJECTIVES: This study investigated the occurrence of extended-spectrum ß-lactamase (ESBL)-producing Salmonella and the associated virulence genes among farmed chickens. METHODS: Cloacal swab samples were collected from apparently healthy and diseased chickens and were cultured for Salmonella using conventional methods. The isolates were serotyped using slide agglutination tests and were examined by polymerase chain reaction (PCR) for the virulence genes invA, stn, svpC and pefA and the outer membrane protein-encoding genes ompA and ompF. Screening for ESBL resistance was performed using the disk-diffusion test, the combinational-disk test with clavulanic acid, and multiplex PCR for blaTEM, blaSHV, blaCTX-M and blaOXA. The presence of the AmpC blaCMy-2 was tested among the ESBL-negative isolates by uniplex PCR. The resistant isolates were partially sequenced based on the stn gene. RESULTS: The Salmonella isolation rate was 3.4% (6/175) from healthy and 11.1% (14/126) from diseased chickens. The 20 isolates belong to serotypes with public health significance like Typhimurium, Kentucky and Infantis. All the isolates possess invA, stn, svpC and ompF genes; 16 isolates harboured ompA, and one carried pefA. Of the 20 isolates, 19 were resistant to more than one antibiotic. Of these 19 isolates, 16 were ESBL-producing with the majority carrying blaTEM and blaSHV genes. The four ESBL-negative isolates carried blaCMY-2. Partial-stn-sequencing of the isolates revealed a high genetic relatedness to Salmonella strains from patients in Egypt and Asia. CONCLUSIONS: Virulent ESBL-producing Salmonella was isolated from healthy and diseased chickens; the strains have a close relationship to human strains, posing a public health threat.

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.

Occurrence of ampicillin-resistant Enterococcus faecium carrying esp gene in pet animals: An upcoming threat for pet lovers.

OBJECTIVES: This study was carried out to investigate oral colonisation by Enterococcus faecalis and Enterococcus faecium in pet dogs and cats, with special reference to antibiotic resistance. METHODS: Oral swabs were collected from 63 pet dogs and 57 pet cats with no known history of hospitalisation. All samples were enriched in Kenner Fecal (KF) broth before being cultured on KF agar to isolate enterococci. E. faecalis and E. faecium were identified by biochemical and molecular techniques. Antimicrobial resistance was determined by the disk diffusion method, and ampicillin-resistant strains were further examined by PCR to detect the esp gene. RESULTS: Oral prevalence rates of E. faecalis among pet dogs and cats were 3.2% and 5.3%, respectively, whilst those for E. faecium were 22.2% and 15.8%, respectively. None of the isolated enterococci were resistant to vancomycin. However, ampicillin-resistant E. faecium (AREfm) was detected in the examined dogs and cats at rates of 14.3% and 5.3%, respectively. Moreover, among the isolated enterococci, six isolates showed multidrug resistance (all AREfm). Whilst the esp gene was detected in only two of nine canine AREfm isolates (multidrug-resistant strains), none of feline AREfm isolates harboured esp. CONCLUSIONS: The occurrence of AREfm and the esp gene among oral isolates from pet dogs and cats represents a great public health hazard for pet owners and highlights possible zoonotic transmission of such a nosocomial pathogen outside healthcare facilities.

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.

The burden of Coxiella burnetii among aborted dairy animals in Egypt and its public health implications.

Q fever is a zoonotic disease of mounting public health implications. Dairy animals are major reservoir for such disease whereas abortion is the main clinical outcome. The current study was conducted to investigate the burden of C. burnetii abortions among dairy animals in Egypt to provide more knowledge for better control of such disease. For this purpose, placental cotyledons and vaginal discharges from 108 aborted dairy animals (27 sheep, 29 goats, 26 cattle, 26 buffaloes) were examined for the presence of C. burnetii by nested PCR. Serum samples from 58 human contacts were examined for the presence of C. burnetii IgG antibodies using ELISA. Out of the 108 examined animals only one goat yielded positive result in both placental tissue and vaginal discharges with an overall prevalence 0.9% while that among goats is 3.4%. Moreover, the seroprevalence of C. burnetii IgG antibodies among the examined individuals was 19% whereas the prevalence in farmers is significantly higher than that among veterinarians and veterinary assistants. In conclusion, C. burnetii may play a role in dairy goat abortions rather than other dairy animals in Egypt while its public health implications cannot be ruled out.

Norway rat (Rattus norvegicus) as a potential reservoir for Echinococcus granulosus: A public health implication.

Cystic hydatidosis is a re-emerging parasitic zoonosis with worldwide distribution. The current study was carried out to investigate the possible role of rats in the epidemiology of such disease in urban and suburban areas. For this purpose, a total of 50 feral Norway rats (Rattus norvegicus) were collected from urban and suburban settings, Cairo, Egypt. Rats were examined to be infected with cystic hydatidosis through serological examination by IHA test as well as post-mortem examination of internal organs, histopathological or molecular identification of the collected cysts. Moreover, 42 persons inhabiting suburban areas were tested for cystic hydatidosis by IHA. The overall seroprevalence rates of cystic hydatidosis in the examined rats and persons were 36% and 11.9% respectively. Cysts from 3 rats were identified as E. granulosus hydatid cysts (one via histopathological examination while the others by molecular technique and genotyped as G6 strain). The results of the current study highlight the possible role of Norway rat in the epidemiological cycle of E. granulosus especially in urban and suburban settings.

Evidence of Zoonotic Transmission of Helicobacter canis Between Sheep and Human Contacts.

Helicobacter species are newly emerging bacteria with great public implications but till now its epidemiology is not fully understood; so, this study was conducted to investigate the possible role of ruminants in the epidemiology of these pathogens. For this purpose, fecal samples were collected from 149 animals (76 sheep, 33 goats, 21 cattle, and 19 buffaloes) and stool specimens from 10 animal caretakers in intimate contact with the examined animals. All samples were examined for the presence of Helicobacter species through detection of Helicobacter genus specific 16S rRNA using PCR. Then, all positive Helicobacter spp. amplicons were sequenced to recognize their species through BLAST analysis at GenBank. The overall prevalence of Helicobacter spp. was 14.8% while the distribution among the different animals was 26.3%, 3%, 4.8%, and 0% in sheep, goats, cattle, and buffaloes respectively. Helicobacter canis was the predominant species and detected only in sheep (21%) and goats (3%). Moreover, Helicobacter winghamensis and Helicobacter canadensis were also detected in sheep but not in other animals, whereas the only positive bovine sample was identified as Helicobacter bovis. On the other hand, 4 out of 10 humans were positive for Helicobacter spp. and all sequences were identified as H. canis. The sequences identity matrix and phylogenetic analysis of H. canis sequences from humans and sheep contacts revealed that one human sequence was identical to that of sheep and making sister group clade, which prove the zoonotic transmission of this pathogen between sheep and human contacts. However, our findings highlight sheep as a potential reservoir for H. canis, further researches are needed to address the potential role of sheep in the food-borne transmission of such emerging pathogen.

Molecular detection of Mycobacterium tuberculosis in cattle and buffaloes: a cause for public health concern.

Tuberculosis is a re-emerging disease causing a growing public health burden. The current study was conducted to investigate the occurrence of Mycobacterium tuberculosis among cattle and buffaloes with tuberculous lesions. Typical tuberculous lesions were collected from 34 cattle and 34 buffaloes (Bubalus bubalis) through postmortem examination of slaughtered animals in abattoirs. DNAs were extracted from samples, and M. tuberculosis was identified by PCR. Positive samples were examined for resistance against rifampicin and isoniazid using GenoType MTBDRplus. Moreover, sera from 90 slaughterhouse workers, butchers, or meat inspectors were examined for the presence of M. tuberculosis antibodies using ELISA. Five cattle (14.7 %) and three buffaloes (8.8 %) tested positive. M. tuberculosis from one cattle was resistant to rifampicin and another was resistant to isoniazid. In addition, the seroprevalence of M. tuberculosis IgG among examined humans was 5.6 %. The occurrence of M. tuberculosis in cattle and buffaloes is a public health concern.

Occurrence of human pathogenic Clostridium botulinum among healthy dairy animals: an emerging public health hazard.

The current study was conducted to investigate the occurrence of human pathogenic Clostridium botulinum in the feces of dairy animals. Fecal samples were collected from 203 apparently healthy dairy animals (50 cattle, 50 buffaloes, 52 sheep, 51 goats). Samples were cultured to recover C. botulinum while human pathogenic C. botulinum strains were identified after screening of all C. botulinum isolates for the presence of genes that encode toxins type A, B, E, F. The overall prevalence of C. botulinum was 18.7% whereas human pathogenic C. botulinum strains (only type A) were isolated from six animals at the rates of 2, 2, 5.8, and 2% for cattle, buffaloes, sheep, and goats, respectively. High fecal carriage rates of C. botulinum among apparently healthy dairy animals especially type A alarm both veterinary and public health communities for a potential role which may be played by dairy animals in the epidemiology of such pathogen.

The zoonotic potential of Giardia intestinalis assemblage E in rural settings.

Giardiasis is a globally re-emerging protozoan disease with veterinary and public health implications. The current study was carried out to investigate the zoonotic potential of livestock-specific assemblage E in rural settings. For this purpose, a total of 40 microscopically positive Giardia stool samples from children with gastrointestinal complaints with or without diarrhea were enrolled in the study as well as fecal samples from 46 diarrheic cattle (18 dairy cows and 28 calves). Animal samples were examined by sedimentation method to identify Giardia spp., and then, all Giardia positive samples from human and animals were processed for molecular detection of livestock-specific assemblage E through amplification of assemblage-specific triosephosphate isomerase (tpi) gene using nested polymerase chain reaction (PCR). The results of the study revealed high unexpected occurrence of assemblage E among human samples (62.5 %), whereas the distribution among patients with diarrhea and those without was 42.1 and 81 %, respectively. On the other hand, the prevalence of Giardia spp. among diarrheic dairy cattle was (8.7 %), while only calves yielded positive results (14.3 %) and all bovine Giardia spp. were genetically classified as Giardia intestinalis assemblage E. Moreover, DNA sequencing of randomly selected one positive human sample and another bovine one revealed 100 and 99 % identity with assemblage E tpi gene sequences available at GenBank after BLAST analysis. In conclusion, the current study highlights the wide dissemination of livestock-specific assemblage E among humans in rural areas, and thus, zoonotic transmission cycle should not be discounted during the control of giardiasis in such settings.