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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.
Показать больше [+] Меньше [-]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.
Показать больше [+] Меньше [-]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.
Показать больше [+] Меньше [-]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.
Показать больше [+] Меньше [-]Potential determinants of Clostridium spp. occurrence in Polish silage
2020
Goldsztejn, Magdalena | Grenda, Tomasz | Kozieł, Nina | Sapała, Magdalena | Mazur, Małgorzata | Sieradzki, Zbigniew | Król, Beata | Kwiatek, Krzysztof
Silage quality deteriorates with Clostridium spp. contamination, and if consumed, such silage jeopardises herd health and productivity. Minimising its occurrence reduces economic and animal welfare risks. The study investigated the influence of environmental and technological determinants on the Clostridium genus’ occurrence in silage. Analyses were conducted on 305 silage samples directly collected from farms located in all Polish provinces. Cultures and isolates were evaluated phenotypically and examined for occurrence of Clostridium spp., particularly C. perfringens and C. botulinum using PCR techniques. The results were statistically analysed using the ᵡ² test for continuous and Student’s t-test for non-continuous values. The most influential effect on Clostridium spp. occurrence is exerted by factors potentially associated with primary production, like the type of fertilisation and the contamination level of the ensiled feed material. Clostridium spp. was detected in 232 (76%) samples, and C. perfringens strains, predominantly toxinotype A, in 79 (26%). C. botulinum occurrence was not detected. Deterioration of silage by clostridia could be prevented by a properly conducted ensiling process with the addition of starter cultures, but the presence of spores mainly depends on primary production and the extent of contamination of the feed material.
Показать больше [+] Меньше [-]Evaluation of a canine transmissible venereal tumour cell line with tumour immunity capacity but without tumorigenic property
2019
Zayas, Yareellys Ramos | Molina, Moisés Armides Franco | Guerra, Reyes Tamez | Padilla, Cristina Rodríguez
Introduction: Canine transmissible venereal tumour (CTVT) is a sexually transmitted tumour affecting dogs worldwide, imposing a financial burden on dog owners. A stable culture cell line in continuous passages for >18 months has only been achieved once. The present study investigated a stable CTVT cell line isolated from a bitch and its potential as a vaccine. Material and Methods: A biopsy from a 2-year-old mongrel bitch with CTVT was obtained for histopathological confirmation and isolation of tumour cells. The isolated cells were cultured to passage 55 and characterised by flow cytometry, with karyotyping by GTG-banding and by PCR detection of myc S-2 and LINE AS1. The isolated CTVT cell line was also used as a preventive vaccine in a canine model. Results: Histopathological analysis of the isolated tumour cells revealed typical CTVT characteristics. Constant proliferation and stable morphological characteristics were observed during culture. Phenotypic analysis determined the expression of HLA-DR⁺, CD5.1⁺, CD14⁺, CD45⁺, CD83⁺, CD163⁺, and Ly-6G-Ly-6C⁺. GTG-banding revealed a mean of 57 chromosomes in the karyotype with several complex chromosomal rearrangements. LINE-c-myc insertion in the isolated CTVT cell line at 550 bp was not detected. However, a 340-bp band was amplified. Isolated CTVT cell line inoculation at a concentration of 1×10⁸ did not induce tumour growth in bitches, nor did a challenge with primary CTVT cells. Conclusion: The present study successfully identified and isolated a stable CTVT cell line that may be useful in CTVT prevention.
Показать больше [+] Меньше [-]New insight of apparently healthy animals as a potential reservoir for Clostridium perfringens: a public health implication
2018
Hamza, Dalia | Dorgham, Sohad M. | Elhariri, Mahmoud | Elhelw, Rehab | Ismael, Elshaimaa
Introduction: Clostridium perfringens is commonly found in the gastrointestinal tract of animals and humans and continues to cause one of the most prevalent foodborne diseases in man. Material and Methods: A total of 355 samples were examined for the occurrence of C. perfringens: rectal swabs from cattle, sheep, and goats, fresh stool samples from diarrhoea sufferers having been in contact with these animals, irrigation water and soil samples from the husbandry sites, and preharvesting fresh produce from farms irrigated with the sampled water. All samples were collected from Cairo and Giza governorates, Egypt. PCR analysis was carried out with positive isolates using the α-toxin gene. Sequence analysis of the gene of C. perfringens isolates was performed using the neighbour-joining approach. Bootstrap analysis was executed with 1,000 resamplings. Results: 174 C. perfringens strains were isolated with a 49.01% prevalence. The highest prevalence of C. perfringens in apparently healthy animals was found in sheep (65.45%) followed by goats (58%), buffaloes (55%), and cattle (47.1%). Its prevalence in humans being in contact with these animals was 47.5%. The bacterium’s isolation from the soil and irrigation water was achieved in 40% and 31.7% of samples, respectively, posing a risk, particularly when the water and soil contact food in the field, shown by the fresh produce isolation of 40%. A significant relationship between the prevalence of C. perfringens in animal and environmental samples was identified (P < 0.05). A significant relationship was identified neither between animal species and C. perfringens prevalence, nor between the environmental source and C. perfringens prevalence (P > 0.05). All isolates were positive for the α-toxin gene by PCR. The sequence analysis and the phylogenetic relationship of the α-toxin genes from different samples revealed that C. perfringens from faeces of apparently healthy cattle, buffaloes, sheep, and goats is a significant threat in places where it can contaminate the soil and water. In addition, the sequence of C. perfringens from humans suffering from diarrhoea was found in the same cluster with the sequence from cows, goats, and sheep. Conclusion: The role of apparently healthy animals in transmitting C. perfringens to humans, either through being in direct or indirect contact via water or soil in the cultivation of vegetables and fruits, was demonstrated.
Показать больше [+] Меньше [-]Genetic analysis of Toxocara spp. in stray cats and dogs in Van province, Eastern Turkey
2018
Oguz, Bekir | Ozdal, Nalan | Serdar Deger, M.
Toxocara canis and Toxocara cati are roundworms of dogs and cats. The purpose of this study was to investigate the infection caused by these ascarids in cats and dogs, using microscopic and molecular analysis methods. Adult ascarids were gathered from the faeces of dogs and cats in Van province, in 2015–2016. Existing keys and PCR sequencing of the ITS-2 fragment were used to identify the morphological features of the parasite species. It was observed that out of 20 adult ascarids, 17 and 3 were found to be Toxocara canis and Toxocara cati, respectively. The ITS-2 gene region was amplified by PCR to perform molecular analysis. Genotyping indicated that the dogs and cats were infected with T. canis and T. cati, respectively, and none had Toxascaris leonina. To the best of our knowledge, this is the first report on the molecular characteristics of adult ascaridoid nematodes from cats and dogs in Turkey. The molecular approaches established in this study enable molecular identification and genetic structure studies of the ascaridoids.
Показать больше [+] Меньше [-]Comparison of two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples
2016
Budniak, Sylwia | Kędrak-Jabłońska, Agnieszka | Szczawińska, Anna | Reksa, Monika | Krupa, Marek | Szulowski, Krzysztof
Introduction: The aim of the study was to optimise and compare two multiplex PCR assays for the detection of Listeria spp. and Listeria monocytogenes in biological samples including the liver, brain, and blood. Material and Methods: Three strains of L. monocytogenes and single strains of each of the species: L. ivanovii, L. innocua, L. grayi, L. welshimeri, and L. seeligeri were used. Additionally, five other species of bacterium were used to evaluate the specificity of the tests. Results: Specific amplification products were obtained for both multiplex PCR assays, which confirmed the tested strains as Listeria spp. and L. monocytogenes, respectively. Isolates of other species did not yield PCR products. Conclusion: Both multiplex PCR assays proved to be significantly sensitive and highly-specific methods for the detection of Listeria strains.
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