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Molecular identification and antifungal susceptibility patterns of dermatophytes isolated from companion animals with clinical symptoms of dermatophytosis Full text
2021
Katiraee, Farzad | Kouchak Kosari, Yasaman | Soltani, Minoo | Shokri, Hojjatollah | Hassan Minooieanhaghighi, Mohammad
Molecular identification and antifungal susceptibility patterns of dermatophytes isolated from companion animals with clinical symptoms of dermatophytosis Full text
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.
Show more [+] Less [-]Molecular identification and antifungal susceptibility patterns of dermatophytes isolated from companion animals with clinical symptoms of dermatophytosis Full text
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.
Show more [+] Less [-]Cytogenetic karyotype analysis in selected species of the Erinaceidae family Full text
2019
Grzesiakowska, Anna | Baran, Przemysław | Kuchta-Gładysz, Marta | Szeleszczuk, Olga
Cytogenetic karyotype analysis in selected species of the Erinaceidae family Full text
2019
Grzesiakowska, Anna | Baran, Przemysław | Kuchta-Gładysz, Marta | Szeleszczuk, Olga
Comparative analysis of the karyotype structure was made in two hedgehog species: the northern white-breasted hedgehog (Erinaceus roumanicus) and the African pygmy hedgehog (Atelerix albiventris). The cytogenetic analysis used differential staining techniques (DAPI, Ag-NOR, and C-banding/DAPI) and sequential QFQ/FISH banding with NOR20 and TEL20 probes which showed 45S rDNA and (TTAGGG)ₙ repeat sequences, respectively, on hedgehog chromosomes. It was confirmed that the somatic cells of the hedgehogs have a constant number of chromosomes (2n = 48,XY). Differences were observed in the NOR number between the species. NORs were identified on three autosome pairs in the northern white-breasted hedgehog and on only two pairs in the African pygmy hedgehog. Chromosome analysis by C-banding/DAPI showed large segments of heterochromatin rich in A-T pairs on three autosome pairs in both the northern white-breasted and African pygmy hedgehogs. The heterochromatin segments encompassed large fragments of the longer arm of chromosome pairs 13, 14 and 20. The (TTAGGG)ₙ repeat sequences on the hedgehog chromosomes were only observed in the terminal position of all the chromosomes in both species. Our observations provide new information on the level of diversity within the Erinaceidae family.
Show more [+] Less [-]Cytogenetic karyotype analysis in selected species of the Erinaceidae family Full text
2019
Grzesiakowska Anna | Baran Przemysław | Kuchta-Gładysz Marta | Szeleszczuk Olga
Comparative analysis of the karyotype structure was made in two hedgehog species: the northern white-breasted hedgehog (Erinaceus roumanicus) and the African pygmy hedgehog (Atelerix albiventris).
Show more [+] Less [-]Epidemiology and antibiogram of Riemerella anatipestifer isolated from waterfowl slaughterhouses in Taiwan Full text
2019
Chang, Fei-Fei | Chen, Chang-Chieh | Wang, Shao-Hung | Chen, Chiou-Lin
Epidemiology and antibiogram of Riemerella anatipestifer isolated from waterfowl slaughterhouses in Taiwan Full text
2019
Chang, Fei-Fei | Chen, Chang-Chieh | Wang, Shao-Hung | Chen, Chiou-Lin
Introduction: Laryngeal swab samples collected from three waterfowl slaughterhouses in central Taiwan were cultured and suspected isolates of Riemerella anatipestifer were identified by API 20NE and 16S rDNA PCR. Material and Methods: Serum agglutination was used for serotyping, and antimicrobial susceptibility was tested. Results: Seventy-six R. anatipestifer isolates were detected, and the prevalences in the ducks and geese were 12.3% (46/375) and 8.0% (30/375), respectively. The positive isolation rates were 65.6% for all arriving waterfowl, 76.0% for birds in the holding area, 1.6% for defeathered carcasses, but zero for degummed carcasses. A PCR examination detected R. anatipestifer in the slaughtering area frequently. Serotype B was dominant in both duck (34.8%) and goose (46.7%) isolates, but the wide serotype distribution may very well impede vaccination development. All isolates were resistant to colistin, and 79.7% were resistant to more than three common antibiotics. Conclusion: The results proved that most ducks had encountered antibiotic-resistant R. anatipestifer in rearing, which suggests that the bacterium circulates in asymptomatic waterfowl. It is worth noting that most waterfowl farms were found to harbour R. anatipestifer, and contaminated slaughterhouses are a major risk factor in its spread. Effective prevention and containment measures should be established there to interrupt the transmission chain of R. anatipestifer.
Show more [+] Less [-]Epidemiology and antibiogram of Riemerella anatipestifer isolated from waterfowl slaughterhouses in Taiwan Full text
2019
Chang Fei-Fei | Chen Chang-Chieh | Wang Shao-Hung | Chen Chiou-Lin
Introduction: Laryngeal swab samples collected from three waterfowl slaughterhouses in central Taiwan were cultured and suspected isolates of Riemerella anatipestifer were identified by API 20NE and 16S rDNA PCR.
Show more [+] Less [-]Taxonomic justification of the pathogenic strongylid infecting the Arabian camel Camelus dromedarius as Haemonchus longistipes by morphological and molecular phylogeny Full text
2019
Dajem, Saad | Morsy, Kareem | Al-Kahtani, Mohammed | Abdel-Gaber, Rewaida
Taxonomic justification of the pathogenic strongylid infecting the Arabian camel Camelus dromedarius as Haemonchus longistipes by morphological and molecular phylogeny Full text
2019
Dajem, Saad | Morsy, Kareem | Al-Kahtani, Mohammed | Abdel-Gaber, Rewaida
Introduction: There is still lack of morphological and phylogenetic information on the pathogenic nematode of the camel Haemonchus longistipes. In the present study, this parasite was isolated in Saudi Arabia and described. Material and Methods: The abomasa of two Arabian camels were collected from a slaughterhouse in Abha province and examined for nematode infection. Worms were described morphologically and morphometrically by electron microscopy. Multiple sequence alignment and the phylogenetic tree of the parasite were constructed from maximum likelihood analysis of its ITS-2 rDNA sequences. Results: These nematodes had a slender body terminating anteriorly at a conspicuous dorsal lancet. A pair of lateral cervical papillae distant from the anterior end was observed. The buccal aperture was hexagonal and surrounded by two amphids, six externo-labial papillae, and four cephalic papillae. Males terminated posteriorly at a bursa supported by spicules and lateral and dorsal rays. Females were linguiform and knobbed morphotypes with distinct ovijectors and a dorsal rim covering the anal pore. The taxonomy was confirmed by the morphology and number of the longitudinal cuticular ridges in a 43–46 range. The sequence alignment and phylogeny revealed 92% homology with H. longistipes (AJ577461.1), and the sequence was deposited into GenBank. Conclusion: The present study describes H. longistipes morphologically and molecularly which facilitates further discrimination of this species worldwide.
Show more [+] Less [-]Taxonomic justification of the pathogenic strongylid infecting the Arabian camel Camelus dromedarius as Haemonchus longistipes by morphological and molecular phylogeny Full text
2019
Dajem Saad | Morsy Kareem | Al-Kahtani Mohammed | Abdel-Gaber Rewaida
Introduction: There is still lack of morphological and phylogenetic information on the pathogenic nematode of the camel Haemonchus longistipes. In the present study, this parasite was isolated in Saudi Arabia and described.
Show more [+] Less [-]Morphology and molecular study of Fascioloides magna – a growing threat to cervids (Cervidae) in Poland Full text
2016
Houszka, Marek | Piekarska, Jolanta | Podkowik, Magdalena | Gorczykowski, Michał | Bania, Jacek
Morphology and molecular study of Fascioloides magna – a growing threat to cervids (Cervidae) in Poland Full text
2016
Houszka, Marek | Piekarska, Jolanta | Podkowik, Magdalena | Gorczykowski, Michał | Bania, Jacek
Introduction: The giant liver fluke, Fascioloides magna, has spread across Europe over the years posing a serious threat to the Polish cervid population. Material and Methods: Macroscopic and histopathological studies of the liver of 22 roe deer (Capreolus capreolus), 10 red deer (Cervus elaphus), and 6 fallow deer (Dama dama) were performed. Species determination of the recovered liver flukes and eggs was performed by PCR protocol amplifying fragments of ribosomal DNA (ITS2), according to a standard method. Results: The presence of F. magna was confirmed in three (13.6%) roe deer, seven (70.0%) red deer, and two (33.3%) fallow deer. The fluke eggs were found only in the stools of five red deer and one fallow deer. Conclusion: This study presents detailed pathological and histopathological changes in the liver of wild Polish cervids, including roe deer, which were subjected to such study for the first time. The hepatic lesions typical for different stages of liver cirrhosis varied depending on the host species and stage of the disease.
Show more [+] Less [-]Morphology and molecular study of Fascioloides magna – a growing threat to cervids (Cervidae) in Poland Full text
2016
Houszka Marek | Piekarska Jolanta | Podkowik Magdalena | Gorczykowski Michał | Bania Jacek
Introduction: The giant liver fluke, Fascioloides magna, has spread across Europe over the years posing a serious threat to the Polish cervid population. Material and Methods: Macroscopic and histopathological studies of the liver of 22 roe deer (Capreolus capreolus), 10 red deer (Cervus elaphus), and 6 fallow deer (Dama dama) were performed. Species determination of the recovered liver flukes and eggs was performed by PCR protocol amplifying fragments of ribosomal DNA (ITS2), according to a standard method. Results: The presence of F. magna was confirmed in three (13.6%) roe deer, seven (70.0%) red deer, and two (33.3%) fallow deer. The fluke eggs were found only in the stools of five red deer and one fallow deer. Conclusion: This study presents detailed pathological and histopathological changes in the liver of wild Polish cervids, including roe deer, which were subjected to such study for the first time. The hepatic lesions typical for different stages of liver cirrhosis varied depending on the host species and stage of the disease.
Show more [+] Less [-]First record of wild boar infected with Trichinella pseudospiralis in Poland Full text
2016
First record of wild boar infected with Trichinella pseudospiralis in Poland Full text
2016
Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa) in the most popular hunting region of the West Pomeranian Province of Poland.Material and Methods: The Trichinella larvae were identified by digestion method. For species identification of the larvae, multiplex PCR was used according to the European Reference Laboratory for Parasites Multiplex PCR protocol. The results were confirmed by molecular amplification of 5S rDNA gene and sequence analysis.Results: Prevalence of 0.54% Trichinella–positive wild boars in the West Pomeranian Province was recorded. Examination of the larvae showed the occurrence of T. spiralis in 79 %, T. britovi in 16.5 %, mixed infection with T. spiralis/T. britovi in 3.5%, and T. pseudospiralis in 1.0% of the boars.Conclusion: This is the first record of wild boar infected with non-encapsulated larvae of T. pseudospiralis in Poland. The species is very difficult to determine, especially using trichinoscopic method. The discovery of the larvae in the animals which may be intended for human consumption confirms that digestion technique should be the only method used for the inspection of meat, especially that from wild boars..
Show more [+] Less [-]First record of wild boar infected with Trichinella pseudospiralis in Poland Full text
2016
Bilska-Zając Ewa | Różycki Mirosław | Chmurzyńska Ewa | Karamon Jacek | Sroka Jacek | Antolak Ewelina | Próchniak Marek | Cencek Tomasz
Introduction: The paper describes identification of Trichinella species isolated from wild boars (Sus scrofa) in the most popular hunting region of the West Pomeranian Province of Poland.
Show more [+] Less [-]Differentiation of Haemobartonella canis and Mycoplasma haemofelis on the basis of comparative analysis of gene sequences Full text
2002
Birkenheuer, Adam J. | Breitschwerdt, Edward B. | Aleman, A Rick | Pitulle, Christian
Objective-To determine whether Haemobartonella canis and Mycoplasma haemofelis (formerly known as H felis [large form]) can be differentiated by use of comparative analysis of gene sequences. Sample Population-Blood samples obtained from 3 dogs infected with H canis and 2 cats infected with M haemofelis. Procedure-The partial 16S rDNA and ribonuclease P RNA (RNase P) genes were amplified, cloned, and sequenced in blood samples obtained from H canis-infected dogs and M haemofelis-infected cats. The DNA sequences were subjected to comparative analysis. Results-The 16S rDNA sequences of H canis and M haemofelis were nearly identical (homology of 99.3 to 99.7%). In contrast, RNase P gene sequences had a lower degree of sequence homology between the 2 organisms (94.3 to 95.5%). Conclusions and Clinical Relevance-Haemobartonella canis and M haemofelis are not identical organisms. Molecular differentiation of H canis and M haemofelis is more clearly evident by use of comparative analysis of RNase P gene sequences than by comparative analysis of 16S rDNA gene sequences.
Show more [+] Less [-]Assessment of in vitro oxalate degradation by Lactobacillus species cultured from veterinary probiotics Full text
2015
Cho, Jenny G. | Gebhart, Connie J. | Furrow, Eva | Lulich, Jody P.
OBJECTIVE To culture Lactobacillus spp from veterinary probiotics and measure their in vitro oxalate-degrading capacity. SAMPLE 2 commercial veterinary probiotics containing Lactobacillus spp. PROCEDURES Lactobacillus spp were cultured anaerobically on selective deMan, Rogosa, Sharpe agar medium and subcultured for speciation by 16S rDNA gene sequencing. Isolates were inoculated into broth containing sodium oxalate (5 mg/L) and incubated anaerobically for 72 hours. An oxalate-degrading isolate of Lactobacillus acidophilus (American Type Culture Collection [ATCC] 53544) was the positive control sample; sterile broth containing a known quantity of sodium oxalate was the negative control sample. Oxalate concentrations were detected with ion chromatography. Oxalate degradation was assessed with Dunnett tests to detect differences in mean oxalate concentration for each isolate, compared with results for the negative control. RESULTS Lactobacillus acidophilus, Lactobacillus plantarum, and Lactobacillus casei or Lactobacillus zeae (too closely related to differentiate) were isolated from probiotic 1, and L plantarum was isolated from probiotic 2. Sequencing of the 16S rDNA gene confirmed 100% homology to type species. Lactobacillus acidophilus (ATCC 53544) and L acidophilus from probiotic 1 significantly decreased oxalate concentrations by 85.3 and 161.9 mg/L, respectively. Lactobacillus plantarum from probiotics 1 and 2 significantly increased oxalate concentrations by 56.1 and 36.1 mg/L, respectively. Lactobacillus casei did not alter oxalate concentrations. CONCLUSIONS AND CLINICAL RELEVANCE Lactobacillus acidophilus isolates significantly reduced oxalate concentrations. In vivo studies are needed to determine whether probiotics containing L acidophilus decrease urine oxalate concentrations and reduce risk of urolith recurrence in dogs with a history of calcium oxalate urolithiasis.
Show more [+] Less [-]Development of a broad-range quantitative polymerase chain reaction assay to detect and identify fungal DNA in equine endometrial samples Full text
2013
Ferris, Ryan A. | Dern, Katy | Veir, Julia K. | Hawley, Jennifer R. | Lappin, Michael R. | McCue, Patrick M.
Objective: To develop a broad-range 28S ribosomal DNA quantitative PCR (qPCR) assay for detection of fungal DNA in equine endometrial samples. Sample: 12 fungal samples from a clinical diagnostic laboratory and 29 samples obtained from 17 mares. Procedures: The qPCR assay was optimized with commercially acquired fungal organisms and validated with samples obtained from the clinical diagnostic laboratory. Subsequently, 29 samples from 17 mares suspected of having fungal endometritis were evaluated via the qPCR assay and via traditional fungal culture and endometrial cytology. Amplicons from the qPCR assay were subjected to genetic sequencing to identify the organisms. Results: The qPCR assay theoretically had a detection threshold of 2 organisms of Candida albicans. Fungal DNA was amplified from all 12 fungal samples from the commercial diagnostic laboratory. Fungal identification by use of genetic sequencing was successful for 34 of 36 amplicons from the 12 samples assayed. A fungal agent was identified via qPCR assay and genetic sequencing in all 12 samples; in contrast, a fungal agent was identified in only 8 of 12 samples via standard fungal culture and biochemical analysis. The qPCR assay detected fungal DNA in samples from 12 of 17 mares suspected of having fungal endometritis. Conclusions and Clinical Relevance: A rapid, sensitive, and repeatable qPCR assay was developed for detection of fungal DNA from equine endometrial samples. The qPCR may prove to be clinically useful as an adjunct to microbial culture and cytologic examination to provide identification of fungal organisms in a timely manner.
Show more [+] Less [-]Ribotype characterization and antimicrobial susceptibility profiles of Campylobacter coli isolates from swine Full text
2003
Sheffield, C.L. | Hume, M.E. | Droleskey, R.E. | Harvey, R.B. | Bischoff, K.M.
Objective: To evaluate the genetic relatedness and antibiotic sensitivity profiles of Campylobacter coli isolates from sows and piglets housed in an integrated swine production facility. Sample Population: Ninety-nine isolates of Campylobacter coli were collected from 3 sows (Yorkshire-Landrace) and 18 piglets (Yorkshire-Landrace X Duroc or Hampshire) housed in a common farrowing barn. Procedure: When piglets were weaned (21 day of age) fecal samples were collected for the sows and rectal samples were collected from the piglets. Isolation of Campylobacter coli was performed using an enrichment broth and restrictive media under microaerophilic conditions. Results: The Campylobacter coli isolates segregated into 20 ribogroups and exhibited 32 antibiotic susceptibility profiles. The Ribogroup (224-373-S-5) contained 35 isolates from eleven animals. Thirty-eight percent of the animals exhibited a single ribogroup, while 10% of the animals exhibited four ribogroups. No discernible pattern of ribogroup relatedness was observed among the sows and piglets or among littermates. Conclusion: The data suggests a high level of diversity in both ribotypic patterns and antibiotic sensitivity profiles among the Campylobacter coli isolated from related pigs housed in a single facility. Further, no evidence was found for a direct transfer of specific Campylobacter coli ribotypes from a sow to her piglets.
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