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Morphological and Molecular Study on Ctenocephalides Fleas Isolated from Stray Dogs in Tehran
2021
Azadbeh, Sara | Dalimi, Abdolhossein | Jamshidi, Shahram
BACKGROUND: Various flea species have already been reported from dogs, among which the most important ones include Ct. felis, Ct. canis, and P. irritans. Fleas can cause annoyance in dogs and human and transmit a variety of bacterial, fungal, and viral agents to the host. In addition, they could function as an intermediate host of Dipylidium caninum and Hymenolepis diminuta. OBJECTIVES: Due to the lack of molecular species-associated identification data, we conducted the current study to differentiate Ct. felis and Ct. canis with molecular assay. METHODS: In the present study, 605 fleas were primarily collected from the dogs referred to Tehran Veterinary Faculty hospital. Subsequently, the flea species were identified under a microscope with morphological keys. Afterwards, COX1 genes of Ct. felis and Ct. canis were amplified via PCR and the locus was finally compared utilizing RFLP and sequencing. RESULTS: Totally, 605 fleas were isolated from 20 dogs. In morphological studies, three species were identified: Ctenocephalides felis, Ctenocephalides canis, Pulex irritans. Pulex irritans had the highest frequency (61.8 %). In molecular study, 552 bp fragment of COX1 gene in two species was amplified and seen on agarose gel. After sequencing, it was seen that two species sequences in COX1 locus had a similarity of 99 % and all of them depended on Ct. canis. In PCR-RFLP, in which Taq1 enzyme was used for differentiation of two species, the same result was obtained. CONCLUSIONS: Even though these two species of dog flea are distinct morphologically, their molecular differentiation using COX1 genes was not successful.
Show more [+] Less [-]Isolation and detection of antibiotics resistance genes of Escherichia coli from broiler farms in Sukabumi, Indonesia
2021
Aprilia Hardiati | Safika Safika | I Wayan Teguh Wibawan | Agustin Indrawati | Fachriyan Hasmi Pasaribu
Objective: This study aimed to isolate and identify Escherichia coli from broiler samples from Sukabumi, Indonesia. Also, antibiogram studies of the isolated bacteria were carried out consid¬ering the detection of the antibiotic resistance genes. Materials and Methods: Cloaca swabs (n = 45) were collected from broilers in Sukabumi, Indonesia. Isolation and identification of E. coli were carried out according to standard bacterio¬logical techniques and biochemical tests, followed by confirmation of the polymerase chain reac¬tion targeting the uspA gene. Antibiotic sensitivity test, using several antibiotics [tetracycline (TE), oxytetracycline (OT), ampicillin (AMP), gentamicin (CN), nalidixic acid (NA), ciprofloxacin (CIP), enrofloxacin (ENR), chloramphenicol, and erythromycin] was carried out following the Kirby Bauer disk diffusion method. Detection of antibiotic resistance coding genes was carried out by PCR using specific oligonucleotide primers. Statistical analysis was carried out with one-way anal¬ysis of variance. Results: The results showed that 55.6% (25/45) of the samples were associated with the pres¬ence of E. coli. Antibiotic sensitivity test showed that the E. coli isolates were resistant to TE (88%; 22/25), OT (88%; 22/25), AMP (100%; 25/25), CN (64%; 16/25), NA (100%; 22/25), CIP (88%; 22/25), ENR (72%; 18/25), chloramphenicol (0%; 0/25), and erythromycin (92%; 23/25). On the other hand, the antibiotic resistance coding genes were tetA (86.4%; 19/22), blaTEM (100%; 25/25), aac(3)-IV (0%; 0/16), gyrA (100%; 25/25), and ermB (13%; 3/23). It was found that chlor¬amphenicol is markedly different from other antibiotic treatment groups. Conclusion: Escherichia coli was successfully isolated from cloacal swabs of broiler in Sukabumi, Indonesia. The bacteria were resistant to TE, OT, AMP, CN, NA, CIP, ENR, and erythromycin. Chloramphenicol was more sensitive and effective than other antibiotics in inhibiting the growth of E. coli. The antibiotic resistance genes detected were tetA, blaTEM, gyrA, and ermB. [J Adv Vet Anim Res 2021; 8(1.000): 84-90]
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