The aim of this study was to investigate staphylococcal enterotoxins (SEs) by ELISA, and detect the five classical sea, seb, sec, sed, and see genes by real-time PCR in Staphylococcus aureus isolates ...from raw and pasteurized milk samples. Staphylococcus spp. were isolated from 98 out of 100 raw milk samples, and in 6 out of 100 pasteurized milk samples. On further biochemical tests, S. aureus was isolated in 48 samples (48%) of raw milk (n=100) and in one sample (1%) of pasteurized milk (n=100). Ten (10%) out of 100 raw milk samples were positive for at least one enterotoxin, and the most frequently observed SE was SEA (10%), followed by SEE (7%) and SEB (6%), but none of the isolates were positive for SEC and SED. At least one of the SEs gene types (sea, seb, sec, sed, see) was detected in 45 (93.8%; 45/48) S. aureus isolates from raw milk samples. sec, sea, seb, sed, and see genes were observed in 56.2%, 39.5%, 31.2%, 29.1% and 14.5% of strains respectively. The enterotoxin genes were the single type in 21 (46.7%) of the 45 isolates, there were two in 15 (33.3%), three in six (13.3%), four in two (4.4%), and one (2.2%) in all gene regions. The SE gene was not detected in the S. aureus (n=1) isolate from pasteurized milk. As a result of this study, the presence of enterotoxigenic S. aureus in raw milk was revealed, and it was pointed out that these SEs may contribute to cases of staphylococcal foodborne poisoning (SPF).
Boiss. subsp.
(Borbas)
(TS) is a commonly used plant in the treatment of various complaints, including skin wounds in Turkish folk medicine. Despite the widespread traditional use of TS, there is not ...any scientific report confirming the effectiveness of this plant on the healing process. This research aimed to investigate the effects of different extracts obtained from TS on biological events during wound healing, on a cellular basis. In this context, proliferative activities of the extracts, as well as the effects on wound closure and hydroxyproline synthesis, were determined. In addition to wound healing properties, the antioxidant, antibacterial and anti-inflammatory activities of the extracts were evaluated. Decoction (D) and infusion (I) extracts contained the highest amount of phenolic content and showed the most potent activity against DPPH radical. All extracts exhibited complete protection against the damage induced by hydrogen peroxide (H
O
) by increasing cell viability compared to only H
O
-treated groups, both in co-treatment and pre-treatment protocols. None of the extracts exhibited cytotoxic activity, and most of the extracts from the TS stimulated fibroblast proliferation and migration. All TS extracts exert anti-inflammatory activity by suppressing the overproduction of tumor necrosis factor-alpha (TNF-α) and nitric oxide (NO). The most pronounced activity on hydroxyproline synthesis was observed in D extract. In summary, it was observed that TS extracts can promote the healing process by enhancing fibroblast migration, proliferation and collagen synthesis as well as suppressing pro-inflammatory cytokines. The obtained data in this work support the traditional use of TS as a valuable plant-based compound for the treatment of wounds.
This study was designed to investigate in vitro biological activities and phytochemical composition of aqueous and ethanolic extracts from Achillea sintenisii Hub- Mor. (AS). To determine the ...chemical composition of AS extracts, phytochemical analyses were performed by using HPLC–ESI-Q-TOF-MS-MS. Afterwards, both extracts were investigated in terms of their effect on fibroblast proliferation, collagen synthesis, and hydrogen peroxide-induced damage. In addition to cell culture analysis, antibacterial, antioxidant, hyaluronidase inhibitory activities and total phenolic contents of the extracts were analyzed in cell-free systems. Our results demonstrated that the aqueous and ethanolic extracts did not show cytotoxic activity on fibroblasts, on the contrary, promoted fibroblast proliferation. Both AS extracts potently inhibited hyaluronidase activity and the inhibitory effect of ethanolic extract was comparable with the positive control, especially at high concentrations. The aqueous extract was the potent stimulator of collagen synthesis at 200 µg/mL concentration. Although the ethanolic extract showed antibacterial activity against all gram-positive bacteria, the aqueous extract was only effective against K. pneumoniae and B. subtilis. The ability of AS extracts, which have a rich phenolic compound content (≥50 mg GAE/g), to scavenge free radicals and protect fibroblasts against hydrogen peroxide-induced damage can be considered as a result of their antioxidant potential. Our findings scientifically support the widespread use of this plant, by demonstrating the pharmacological properties of the extracts.
The increase of antimicrobial resistance in companion animals has already been reported worldwide; however, there is a deficiency of data and studies focusing on the resistance profiles of ...Escherichia coli isolates from Passeriformes and Psittaciformes which regarded to be among the most common and popular pet/companion bird species. For this reason, the current research was aimed to evaluate the presence of E. coli isolates from apparently healthy companion birds and their antimicrobial resistance profiles.
Two hundred sixty-five cloacal swab samples collected from apparently healthy companion birds (116 parakeets, 59 canaries, 56 parrots, 30 Indian nightingales, 3 finches, and 1 Golden finch) were examined by conventional bacteriological procedures for the identification of E. coli. Susceptibilities against 16 antimicrobials from 8 different classes and extended-spectrum beta-lactamase (ESBL) production was also determined. Moreover, all isolates were analysed by PCR assays for ESBL, Metallo-β-lactamases, serin-carbapenemase, AmpC beta-lactamase, plasmid-mediated quinolone resistance (PMQR), and aminoglycoside resistance genes.
E. coli were isolated from 37.7% of the samples. Majority of the isolates were found resistant to tetracycline (84%) followed by sulfamethoxazole/trimethoprim (46%), streptomycin (34%), and kanamycin (25%). Eleven parakeet and 2 parrot isolates were found resistant to all quinolone class antimicrobial agents, and three parakeet isolates showed resistance to all aminoglycosides. Additionally, 67% of the isolates exhibited multi-resistance, defined herein as 3 or more antimicrobial classes.
PMQR determinants (qnrB and qnrS) were determined in 3 E. coli isolates from a parrot and two parakeets. Furthermore, seven aminoglycoside resistance genes aac(3)-IIa(aacC2), strA, strB, aadA (aadA1 or aadA2), aphA1, aphA2, and ant(2’’)-Ia(aadB) were found in 3 parakeet isolates. Also, the most promising result of this study is that ESBL production was not determined phenotypically and genotypically.
Here, we present the first report that various aminoglycosides and quinolone resistance genes of E. coli isolates from parakeets, and a parrot was present in Turkey. In summary, the consequences of the current research emphasise that the companion birds may act as substantial reservoirs carrying antimicrobial resistance and estimate the potential risk for humans, it is critical to define their role as reservoirs.
Exotic pet animal ownership is on the rise all over the world. Regardless of being companion animals which are important subjects in veterinary medicine, they also play a role in the transmission of ...diseases to other animals and human beings. Therefore, veterinarians are expected to have the knowledge and good practice in exotic pet medicine. This survey was performed among small animal practitioners with the aim of identifying their self-evaluation of competency and knowledge about the exotic pet medicine in Istanbul. As a data collection tool, a three-part questionnaire developed by the researchers was used in the current study. The first part of the survey covered demographic variables of respondents; the second part consisted of 6 questions, which examine the self-evaluation of small animal practitioners on competency and knowledge about the exotic pet medicine practice. The third part consisted of 8 Likert type questions about the husbandry, transmission, prevention, diagnosis and therapy of the diseases for the four different exotic pets including fish, turtle, other reptiles and bird. The results showed that approximately 80% of the respondents consider that exotic pet disease is essential regarding veterinary medicine. Thirty-five percent of the veterinarian said that they were not sure about "what is an exotic pet disease" while, 53% of them responded that question, as they did not have any knowledge about the exotic pet diseases. For the 91.4% of the clinics investigated, the exotic animals as a patient were 1-10% or less than 1% of all the patients. It has shown that 42.4%, 32%, 16.9%, and 8.7% of the exotic pets' species examined were birds, turtles, the other reptiles, and 8.7% fish, respectively. Ninety percent of the veterinarians consider that they did not get enough education about the exotic pet animal practice during their undergraduate study at the Faculty of Veterinary Medicine, Istanbul. It has been determined that more than half of the participants (65%) were considered themselves as having adequate knowledge of the husbandry, transmission, prevention, diagnosis and therapy of the diseases of the birds. However, they did not have enough experience about turtle, other reptiles and fish. As a conclusion, the results indicated the importance of education, specialisation and practice on potential exotic pet species. Keywords: Exotic pets, istanbul, self-evaluation, survey. Veterinary Medicine
This study aimed to develop direct PCR methods, which enable the diagnosis of brucellosis agents from ruminant aborted fetus samples at species and genus levels, and determine the applicability of ...the newly developed methods. For this purpose, 137 lung, 137 liver, and 52 fetal stomach fluid samples belonging to 166 ruminant aborted fetuses (326 samples in total) were examined. Firstly, agent isolation and identification were performed and species-specific multiplex PCR (m-PCR) from the culture was applied to the samples. In addition, the Mayer-Scholl m-PCR method was modified and termed ‘modified Mayer-Scholl’, and genus specific Bcsp31 PCR was also modified with minor changes. Four different methods were applied to direct examination samples and the obtained results were compared. The conventional culture method was set as the standard method to which sensitivities and specificities of the molecular methods were calculated. According to the assessments on the basis of fetus (n = 166), sensitivity and specificity values for modified Mayer-Scholl m-PCR method were 94.11% and 98.76%, and the same indicators for the modified Bcsp31 PCR were 95.29% and 98.76%, respectively. When all organ samples were taken into account (n = 326), sensitivity and specificity values for the modified Mayer-Scholl m-PCR method were 85.38% and 98.06%, and for the modified Bcsp31 PCR, they were 83.62% and 98.06%, respectively. As a result, it was found that the diagnostic power of the tests were ‘high’ when results were evaluated at fetus level. On the other hand, it was found to be ‘clinically useful’ when evaluated at organ level. We concluded that species level identifications can be made through the modified Mayer-Scholl method, which is a direct m-PCR method, with a high diagnostic power by specifying DNAs belonging to Brucella species directly from clinical samples.
•Multiplex PCR is applicable to the direct diagnosis of brucellosis from clinical samples•The sensitivity and specificity values of the PCR are considered as satisfactory•For the diagnosis of brucellosis, first, the fetal stomach fluid should be preferred•Then, lung and liver tissues should be opted for diagnosis, respectively
The aim of this study was to investigate staphylococcal enterotoxins (SEs) by ELISA, and detect the fivee classical sea, seb, sec, sed, and see genes by real-time PCR in Staphylococcus aureus ...isolates from raw and pasteurized milk samples. Staphylococcus spp. were isolated from 98 out of 100 raw milk samples, and in 6 out of 100 pasteurized milk samples. On further biochemical tests, S. aureus was isolated in 48 samples (48%) of raw milk (n=100) and in one sample (1%) of pasteurized milk (n=100). Ten (10%) out of 100 raw milk samples were positive for at least one enterotoxin, and the most frequently observed SE was SEA (10%), followed by SEE (7%) and SEB (6%), but none of the isolates were positive for SEC and SED. At least one of the SEs gene types (sea, seb, sec, sed, see) was detected in 45 (93.8%; 45/48) S. aureus isolates from raw milk samples. sec, sea, seb, sed, and see genes were observed in 56.2%, 39.5%, 31.2%, 29.1% and 14.5% of strains respectively. The enterotoxin genes were the single type in 21 (46.7%) of the 45 isolates, there were two in 15 (33.3%), three in six (13.3%), four in two (4.4%), and one (2.2%) in all gene regions. The SE gene was not detected in the S. aureus (n=1) isolate from pasteurized milk. As a result of this study, the presence of enterotoxigenic S. aureus in raw milk was revealed, and it was pointed out that these SEs may contribute to cases of staphylococcal foodborne poisoning (SPF). Key words: ELISA; staphylococcal enterotoxins (SEs); real-time PCR; staphylococcal foodborne poisoning (SPF) Cilj je bio u izolatima bakterije Staphilococcus aureus iz sirovog i pasteriziranog mlijeka pomocu ELISA testa istraziti enterotoksine staflokoka (SEs) te uporabom PCR-a u stvarnom vremenu provjeriti prisustvo pet gena, sea, seb, sec, sedi i see. Bakterija Staphylococcus spp. izolirana je iz 98 od 100 uzoraka sirovog mlijeka i iz 6 od 100 uzoraka pasteriziranog mlijeka. Daljnjim je biokemijskim testiranjem S. aureus izolirana iz 48 uzoraka (48%) sirovog mlijeka (n=100) i iz jednog uzorka (1%) pasteriziranog mlijeka (n=100). Ukupno je 10 uzoraka (10%) od 100 uzoraka sirovog mlijeka bilo pozitivno na barem jedan enterotoksin, a najcesci je enterotoksin bio SEA (10%), zatim SEE (7%) i SEB (6%). Ni jedan od izolata nije bio pozitivan na SEC i SED. Najmanje je jedan tip gena za enterotoksine (sea, seb, sec, sed, see) otkriven u 45 izolata (93,8%, 45/48) S. aureus iz uzoraka sirovog mlijeka. Gen sec uocen je u 56, 2% sojeva, gen sea u 39,5% sojeva, gen seb u 31,2% sojeva gen sed u 29,1% sojeva i gen see u 14,5% sojeva. Geni za enterotoksine pronadeni su kao pojedinacan tip u 21 od 45 izolata (46,7%), dva od 15 izolata (33,3%), tri od sest izolata (13,3%), cetiri od dva izolata (4,4%) i jedan (2,2%) u svim regijama gena. Gen za enterotoksin nije pronaden u izolatu S. aureus (n=1) iz pasteriziranog mlijeka. Ovo je istrazivanje pokazalo da se enterotoksigena bakterija S. aureus nalazi u sirovom mlijeku, pri cemu je naglaseno da enterotoksini mogu pridonijeti trovanju hranom uzrokovanom staflokokima (SPF). Kljucne rijeci: ELISA; enterotoksini staflokoka (SEs); PCR u stvarnom vremenu; trovanje hranom uzrokovano staflokokima (SPF)
This study, for the first time in Turkey, investigated the existence of Chlamydophila psittaci and determined the prevalence of its disease, chlamydiosis, in pet birds. Polymerase chain reaction ...(PCR) was compared with other testing methods that have been typically used in the diagnosis of C. psittaci. Fecal specimens (n = 96) of avian origin were tested by PCR and two identification methods, modified Gimenez staining (mGS) and direct fluorescein–conjugated monoclonal antibody staining (FA). The identification methods were implemented by staining the yolk sacs of embryonated chicken eggs inoculated at 6 days of age and harvested between 3 and 10 days after inoculation. Fecal specimens from pet birds were randomly collected from pet shops and homes. These specimens were then used to isolate C. psittaci and to detect its specific DNA. The inocula that were prepared from fecal specimens were then inoculated into yolks of 6-day-old embryonated chicken eggs. The preparations from egg yolk sacs were examined with mGS and direct FA after three blind passages. The PCR method was used to detect specific DNA in feces. In 96 fecal specimens, 33 (34.4%) were positive with PCR, 21 (21.9%) were positive with mGS, and 29 (30.2%) were positive with FA. Among 33 positive specimens with PCR, 28 specimens were positive with FA, and 20 specimens were positive with mGS. The sensitivity and specificity were 59% and 94% between FA and mGS, and 97% and 93% between FA and PCR, respectively.
The purpose of the study was to examine the prevalence of Escherichia coli in shrimps and mussels, and to determine the distribution of β-lactam, aminoglycoside, quinolone, and multi-drug resistance ...phenotypically and genotypically in E. coli isolates obtained from mussels and shrimps in Istanbul. Faecal samples were collected from mussels (n = 96) and shrimps (n = 96) from the Marmara Sea coastline and fish markets in Istanbul. For the detection of antibiotic susceptibilities, seven antibiotic groups were used. β-lactamase, aminoglycoside, and quinolone genes were also determined. A total of 34 (17.7%, 15 shrimps, and 19 mussels) E. coli were isolated, and 17 (50%) were found to be resistant to one or more antimicrobials. The highest resistance was seen against aminoglycosides with 11 isolates (32.35%), followed by quinolones with 10 isolates (29.41%) and extended-spectrum β-lactamase (ESBL) with 4 isolates (11.76%). Multi-drug resistance was detected in 5 isolates (14.7%) from 3 shrimp and 2 mussel samples. The prevalence of ESBL genes was demonstrated at 3.84% in mussels and shrimp samples. There were no AmpC and carbapenemase-producing genes. These samples harbored blaCTX-M-1 (n = 3) and blaTEM (n = 4). Ten isolates were resistant to aminoglycosides genotypically. Resistance genes detected were strB in 2 isolates, aadA in 5, strB and aadA together in 3, ANT('')-Ia, aphA1 and aphA2 simultaneously in 3, aphA1 in 1, aac(3)-IIa in 1 isolate. aac(6')-Ib-cr gene was detected in only one of 10 phenotypically resistant isolates to quinolones.
Tetracycline is a broad-spectrum antibiotic which is commonly used in humans and animals for treatment of bacterial infections. Therefore, tetracycline-resistant Enterobacteriaceae species found in ...the nature, humans, and animals are usually considered a serious health concern. The feces of birds that live with humans may be a source for of these antibiotic resistant bacteria. For this reason, presence of tetracycline-resistant Enterobacteriaceae in bird droppings collected from 18 different breeders and pet shops fed in Istanbul was investigated by cultural and molecular methods in terms of the presence of that a/b gene. In addition, susceptibilities of isolates to various antibiotics were also determined. The current study reported that Enterobacter cloacae, Citrobacter freundi, Escherichia coli, Edwardsiella spp., Enterobacter spp., Escherichia vulneris, Klebsiella pneumonia, Pantoea agglomerans, Proteus mirabilis, Pantoe spp., Pseudomonas spp., Serratia marcescens, Salmonella spp., Shigella spp. were isolated from the samples. Ninety-seven of the isolates (97/150) were resistant to tetracycline (65.2%). In addition, the isolates were resistant to other antibiotic medications and 114 of them (58%) evaluated as multi drug resistant. The tet (A) and tet (B) genes were found in bacteria isolated from synantrophic birds’ feces as 46.6% and 8%, respectively. Although working conditions are limited, the results obtained provide baseline information regarding antibiotic resistance to Enterocatericeae isolates obtained from captive birds’ feces. Thus, the results of the present study emphasize the necessity of genotypic resistance research in future studies to help maintain antibiotic treatment efficacy for Enterocatericeae infections.
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