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Identifying Corynebacterium pseudotuberculosis in Sheep of Kurdistan Province in Iran by Culture and Polymerase Chain Reaction and Determining the Antibiotic Resistance of its Isolates
2024
Ataei Kileh Golan, Jamil | Derakhshan, Safora | Sharifi, Aram | Nayeri Fasaei, Bahar | Zahraei Salehi, Taghi
BACKGROUND: Corynebacterium pseudotuberculosis is the etiological agent of caseous lymphadenitis (CLA), a chronic and very common disease in sheep and goats, which can lead to severe economic losses in the livestock industry.OBJECTIVES: This study aims to investigate the prevalence of CLA in sheep in Kurdistan province of Iran using phenotypic and molecular methods, and assess the antibiotic resistance of isolated Corynebacterium pseudotuberculosis.METHODS: In this study, from September to March 2022, 270 samples of skin abscesses were collected from sheep in livestock farms of Kurdistan province. Immediately, using the cold chain system, the samples were transferred to the microbiology laboratory of the Faculty of Medicine at Kurdistan University of Medical Sciences. Identification of isolates was done using biochemical tests and polymerase chain reaction (PCR) method. The antibiotic resistance of the isolates was examined using the Kirby-Bauer disk diffusion method.RESULTS: Based on biochemical tests, out of 270 samples, 82 suspected to have Corynebacterium pseudotuberculosis. Out 82 samples, the presence of bacteria was confirmed in 76 samples by the PCR. The antibiotic sensitivity test showed that the isolates had high sensitivity to doxycycline and ceftriaxone and high resistance to streptomycin and kanamycin.CONCLUSIONS: The CLA has a high prevalence in sheep in Kurdistan province. According to high resistance rate of Corynebacterium pseudotuberculosis to streptomycin and kanamycin, it recommended to avoid treatment of CLA cases with these antibiotics.
Mostrar más [+] Menos [-]Identification of Campylobacter spp. from poultry skin using methods based on bacterial culture and polymerase chain reactions
2016
ebrahimi lagha, fateme | zeynali, fariba | Rezazadeh bari, mahmood
BACKGROUND: Food infections caused by Campylobacter are one of the gastrointestinal inflammations in humans is health and economic losses in the community is important. OBJECTIVES: To determine the prevalence of Campylobacter contamination in chicken skin samples of Urmia, using bacterial culture and polymerase chain reactions. METHODS: 80 samples of chicken skin from the Protein Gostare Sina slaughter house located in the city of Urmia in equal numbers in the winter and spring seasons were collected. The survival of Campylobacter after 24 hours in refrigerated conditions was studied in samples. Positive samples were used for DNA extraction and PCR. To investigate the phylogenetic isolates, positive samples PCR were sequenced. RESULTS: 58/75% of chicken skin using bacterial cultures, Campylobacter were positive. The Results study the survival Campylobacter in cold conditions after 24 hours, showed that no significant decrease in the survival Campylobacter as well as contamination levels were significantly higher in spring than in winter, which may be due to the high temperature of environment that created the favorable conditions for Campylobacter. CONCLUSIONS: Chicken skin is the reservoir of Campylobacter. This issue of public health care and control at all stages of production and supply of poultry products, also the transfer of it to other parts of poultry carcasses should be considered.
Mostrar más [+] Menos [-]Detection of Toxoplasma gondii in feline and canine biological samples by use of the polymerase chain reaction.
1996
Stiles J. | Prade R. | Greene C.
Delayed seroconversion following naturally acquired caprine arthritis-encephalitis virus infection in goats.
1993
Rimstad E. | East N.E. | Torten M. | Higgins J. | DeRock E. | Pedersen N.C.
One hundred eight milking goats from a dairy that had been using a modified caprine arthritis-encephalitis virus (CAEV) eradication program were tested for CAEV antibodies by serologic methods and for proviral CAEV DNA by use of polymerase chain reaction (PCR) technology. All goats were free of clinical symptoms of CAEV infection. Twenty-seven of the 108 goats were considered seropositive, on the basis of ELISA results. Proviral CAEV DNA was detected, using PCR techniques, in mononuclear leukocytes in blood samples obtained from 25 of the these 27 seropositive goats. Twenty of the 81 seronegative goats also had positive PCR test results. Ten of these goats seroconverted by 8 months later, and virus was readily isolated from mononuclear leukocytes in venous blood samples after the goats had seroconverted. Virus was also isolated from mononuclear leukocytes in blood samples collected from 4 of 11 goats that were seronegative, but had positive PCR test results. These results indicated that seroconversion can be delayed for many months following natural infection with CAEV. Delayed seroconversion appears to be a feature of CAEV infection, which may have direct implications for CAEV eradication programs and epidemiologic studies that rely on serologic methods to detect infected goats.
Mostrar más [+] Menos [-]Correlation between clinical signs of feline conjunctivitis and molecular detection of felid herpesvirus-1, feline calicivirus, chlamydophila felis and mycoplasma felis in cats from shelters in Rio de Janeiro
2017
Natasha Baumworcel | Ana Maria Barros Soares | Sheila Bruna Silva | Núbia Karla Oliveira Almeida | Tatiana Xavier de Castro
Objectives: To perform molecular diagnosis of microbial agents (FHV-1, FCV, Mycoplasma felis, and Chlamydophila felis) in kittens with conjunctivitis and correlate the clinical signs with clinical severity. Material and Methods: A total of 108 conjunctival swab were collected from kittens without (G1; n = 40) and with (G2; n = 68) clinical signs of conjunctivitis. Animals from G2 group were scored from 1 (mild) to 4 (severe) according to the severity of conjunctivitis. All samples were submitted to PCR and RT-PCR. Results: FHV-1 was detected in 62/108 (57.4%) of samples, FCV in 40/108 (37.0%), M. felis in 11/108 (10.2%) and C. felis in 26/108 (24.1%). Mixed infections were detected in 39/108 (36.1%). In G1, 28/40 (70.0%) were positive for one or more agents, in G2, 58/68 (85.3%) were positive (P = 0.03). In 1, single infections by FHV-1were found in 21/40 (52.5%) samples, FCV in 2/40 (5.0%), C. felis in 1/40 (2.5%), and no pathogens were detected in 12/40 (30%) of samples, while mixed infections accounted for 29/40 (72.5%) of the cases. In G2, single FHV-1 infections were found in 31/68 (45.6%) samples, FCV in 10/68 (14.7 %), M. felis in 2/68 (3.0%) and C. felis also in 2/68 (3.0%), and no pathogens were detected in 10/68 (14.7%) samples, while mixed infections accounted for 36/68 (52.0%) of the cases. They were categorized as grade 1, 20/68 (29.4%), grade 2, 14/68 (20.6%), grade 3, 21/68 (30.9%) and grade 4, 13/68 (19.1%). The presence of FHV-1 and FCV is equally distributed among the four categories. More severe clinical signs, scores 3 and 4, are related to coinfections by C. felis and M. felis. Conclusions: FHV-1, FCV, C. felis and M. felis were identified in feline conjunctivitis. Co-infections are related to more severe cases of conjunctivitis. Molecular diagnosis is helpful to detect asymptomatic carriers and is a rapid and accurate method to determine the pathogen of feline conjunctivitis.
Mostrar más [+] Menos [-]First molecular assessment of the African swine fever virus status of Ornithodoros ticks from Swaziland
2014
Carin I. Boshoff | Armanda D.S. Bastos | Mzwandi M. Dube | Livio Heath
African swine fever (ASF) is an economically significant haemorrhagic disease of domestic pigs. It is caused by the African swine fever virus (ASFV), a deoxyribonucleic acid (DNA)arbovirus. Argasid ticks of the genus Ornithodoros, which are widely distributed throughout southern Africa, play a primary role in virus maintenance and spread within the endemic sylvatic cycle. The ASF status of Swaziland is unknown, but this land-locked country is surrounded by ASF-positive countries, has a burgeoning pig industry and sylvatic cycle hosts present within its borders. In this first assessment of ASF status, warthog burrows in seven nature reserves and game management areas in Swaziland were investigated for tick and virus presence. Tick infestation rates of between 33.3% – 88.8% were recovered for the four Ornithodoros-infested reserves. A total of 562 ticks were screened for virus genome presence using a duplex Polymerase Chain Reaction (PCR) that targets the C-terminal end of the p72 gene of the ASFV and confirms DNA integrity through amplification of the 16S rRNA tick host gene. All samples were negative for virus genome presence and positive for the tick genome target. Nucleotide sequencing of the latter confirmed that Ornithodoros ticks from Swaziland are identical to those from the Kruger National Park in South Africa across the gene region characterised. Whilst this first evaluation of ASF presence in Swaziland indicates that the virus does not appear to be present in the key virus vector, the presence of sylvatic cycle hosts, together with the country’s proximity to ASF-affected countries calls for expanded investigations and regular monitoring of the ASF status of Swaziland.
Mostrar más [+] Menos [-]Detection of Haemophilus parasuis isolates from South China by loop-mediated isothermal amplification and isolate characterisation
2012
Jian-min Zhang | Hai-yan Shen | Ming Liao | Tao Ren | Li-li Guo | Cheng-gang Xu | Sai-xiang Feng | Hui-ying Fan | Jing-yi Li | Ji-dang Chen | Bin Zhang
Haemophilus parasuis is the etiological agent of Glässer’s disease, which is characterised by fibrinous polyserositis, meningitis and polyarthritis, causing severe economic losses to the swine industry. In this study, a loop-mediated isothermal amplification (LAMP) test was developed to improve the specificity, facility and speed of diagnosis of H. parasuis isolates. The LAMP assay rapidly amplified the target gene within 50 min incubation at 63 °C in a laboratory water bath. The LAMP amplicon could be visualised directly in the reaction tubes following the addition of SYBR Green I dye. The detection limit of this LAMP method was 10 CFU/mL, which was 10 times more sensitive than the earlier 16S rRNA polymerase chain reaction (PCR) test conducted by Oliveira, Galina and Pijoan (2001), and no cross-reactivity was observed from other non-H. parasuis strains. This LAMP test was evaluated further on 187 clinical specimens from pigs suspected of being infected with H. parasuis. Forty-three were found positive by bacterial isolation of H. parasuis, as well as by the 16S rRNA PCR and LAMP tests. The 43 H. parasuis isolates were classified into 9 serovars and had 37 genetic patterns when analysed by pulsed-field gel electrophoresis (PFGE). This displayed that various H. parasuis serovars and genotypes were widely distributed in South China. Therefore, the speed, specificity and sensitivity of the LAMP test, the lack of a need for expensive equipment, and the visual readout showed great potential for a correct clinical diagnosis of H. parasuis in favour of controlling Glässer’s disease.
Mostrar más [+] Menos [-]Use of real-time quantitative reverse transcription polymerase chain reaction for the detection of African horse sickness virus replication in Culicoides imicola
2011
Elisabeth G. Scheffer | Gert J. Venter | Christopher Joone | Nikolaus Osterrieder | Alan J. Guthrie
Despite its important role as vector for African horse sickness virus (AHSV), very little information is available on the dissemination of this virus in Culicoides (Avaritia) imicola Kieffer (Diptera: Ceratopogonidae). This study reports on the applicability of a real-time quantitative reverse transcription polymerase chain reaction (RT-qPCR) to detect AHSV in dissected midges. A total of 96 midges were fed on AHSV-infected blood, after which one test group was dissected into head/thorax and abdomen segments immediately after feeding and the other only after 10 days of incubation. The majority of the midges (96%) ingested the virus successfully and there was no significant difference between the virus concentration in the heads/thoraxes and the abdomens immediately after feeding. After incubation, virus was detected in 51% of the midges and it was confined to the abdomen in the majority of these. The fact that virus was detected only in the heads/thoraxes of four Culicoides midges after incubation suggests the presence of a mesenteronal escape barrier. Replication in the salivary glands was not shown. An increase of the mean virus concentration in the abdomen after incubation indicates localised viral replication. The real-time RT-qPCR is recommended for further studies investigating the replication and dissemination of AHSV in Culicoides midges.
Mostrar más [+] Menos [-]Detection of Chlamydophila psittaci in chickens by complement fixation test and polymerase chain reaction
2007
Wafaa A. Osman | A. L. El-Naggar | Azza S. A. Gooda | Mona A. Mahmoud
This study was carried out on 68 randomly collected chickens located at Ras Sedr Research Station, Desert Research Center, 68 serum samples were examined serologically by complement fixation test (CFT). Twenty out of 68 (29.91%) had antibodies against Chlamydophila psittaci . Ten blood samples of the serologically positive cases were subjected to polymerase chain reaction (PCR) and showed positive results for Chlamydophila psittaci at 119 bp. Therefore PCR was found to be reliable, rapid, sensitive and specific technique for the detection Chlamydophila psittaci in birds. Serologically positive birds did not show any clinical symptoms of disease, but they were in contact with sheep and goat that showed previous abortion and were positive for C. abortus. It is recommended to avoid breeding of chickens with other animal species in the same yard because chickens become asymptomatic carrier with shedding of Chlamydophila psittaci in their feaces and respiratory discharges.
Mostrar más [+] Menos [-]Molecular identification and antifungal susceptibility patterns of dermatophytes isolated from companion animals with clinical symptoms of dermatophytosis
2021
Katiraee, Farzad | Kouchak Kosari, Yasaman | Soltani, Minoo | Shokri, Hojjatollah | Hassan Minooieanhaghighi, Mohammad
Dermatophytosis is a common skin disease in cats and dogs caused by Microsporum and Trichophyton fungi. Species identification and knowledge of their antifungal susceptibility are therapeutically and epidemiologically important. This study assessed the prevalence of feline and canine dermatophytosis in Iran, identified the aetiological agents molecularly and tested their antifungal susceptibility. A total of 308 companion animals (134 dogs and 174 cats) with skin lesions were examined from March 2015 to March 2018. Hair and skin samples were examined by microscopy with 20% KOH and cultured on Sabouraud dextrose agar with cycloheximide and chloramphenicol. Fungal isolates were confirmed by sequencing of the internal transcribed spacer (ITS) r-DNA region. The antifungal susceptibility of dermatophytes was tested by broth microdilution assay using standard drugs. Dermatophytes were found in 130 (42.2%) samples, 62 of them feline and 68 canine. Based on sequencing of all strains, M. canis (78.5%, P<0.05), M. gypseum (10.7%), and T. mentagrophytes (10.7%) were the dermatophytes isolated. The non-dermatophyte species Nannizziopsis vriesii was also isolated from two feline dermatomycosis cases. Dogs and cats younger than one year (61.5%) showed a statistically significantly higher prevalence of infection (P<0.05). Caspofungin produced the lowest geometric mean MIC at 0.0018 μg/mL, followed by ketoconazole, terbinafine, itraconazole, miconazole, griseofulvin, clotrimazole and fluconazole, in a 0.038–1.53 μg/mL range. This is the first molecular study to identify the causes of pet dermatophytosis in north-western Iran. ITS-PCR was shown to be a useful and reliable method for the identification of closely related species of dermatophytes in clinical and epidemiological settings. The lowest MIC of caspofungin indicated that this drug was the most potent in vitro.
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