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1.
J Vet Diagn Invest ; 36(1): 108-111, 2024 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-37919953

RESUMO

Mycoplasma equirhinis is the predominant equine Mycoplasma sp. isolated from clinically normal horses and is suspected to be associated with inflammatory airway disease in which cough is the primary sign. Quantitative evaluation of bacterial counts is useful in assessing the association between the bacteria in samples and observed clinical signs, but this evaluation has been difficult with conventional culture methods of M. equirhinis given the need for pre-enrichment using liquid cultures. We established a quantitative real-time PCR (qPCR) assay for the quantification of M. equirhinis, targeting the hypothetical protein FJM08_00025. We confirmed its high species-specificity for M. equirhinis and a limit of detection of 2.9 copies/reaction. We quantified M. equirhinis in tracheal wash samples from 20 clinically normal horses and 22 coughing horses. The copy numbers detected by qPCR in 18 of the 22 samples from clinically affected horses were within the range detected in the 20 clinically normal horses (0-84 copies/reaction). The remaining 4 samples had considerably higher copy numbers (734-1,620,000 copies/reaction), suggesting the likely involvement of M. equirhinis infection. Quantitative evaluation of M. equirhinis over time using our qPCR assay may allow a more accurate assessment of M. equirhinis infection in coughing horses compared to culture methods.


Assuntos
Doenças dos Cavalos , Mycoplasma , Cavalos , Animais , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Reação em Cadeia da Polimerase em Tempo Real/métodos , Mycoplasma/genética , Traqueia/microbiologia , Doenças dos Cavalos/diagnóstico , Doenças dos Cavalos/microbiologia
2.
Jpn J Radiol ; 42(2): 174-181, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-37815695

RESUMO

OBJECTIVE: This study aims to retrospectively evaluate the outcomes of uterine artery embolization (UAE) for uterine fibroids (UFs), specifically submucosal UFs, according to the International Federation of Gynecology and Obstetrics (FIGO) classification of UFs. MATERIALS AND METHODS: Forty-two patients with symptomatic UFs underwent UAE with Embosphere® between July 2016 and November 2021. MRI was performed before, at 3 and 6 months after the UAE. At each examination, the volume of UF was measured, and the percentage volume reduction rate (VRR) was calculated. The technical success rate (TSR), symptom improvement rate (SIR), regrowth rate (RR) after 6 months, and adverse events (AEs) were examined; VRR was compared between patients with submucosal UFs (FIGO types 0-2, group A), those with submucosal contacts (FIGO type 3, group B), and those without submucosal UFs (FIGO types 4-7, group C). Statistical analysis was performed on the difference in VRR between groups A, B, and C at 3 and 6 months after UAE. The relationship with hormone levels before UAE and VRR was evaluated. RESULTS: Thirty-seven of the 42 patients were evaluated. Overall, VRR was 37.0% at 3 months and 52.1% at 6 months; TSR, SIR, and RR were 100%, 95.2%, and 5.4%, respectively; VRR at 6 months was 80.7% for group A (n = 7), 57.8% for group B (n = 13), and 37.1% for group C (n = 17). Significant differences were found between A and C (p < 0.001) and B and C (p = 0.023). Hormone levels before UAE had no effect on VRR. There was no significant AEs other than grade 3 pulmonary embolism in one patient. CONCLUSION: UAE was effective for submucosal FIGO types 0-3. UAE was especially useful as an option for FIGO type 3 with a low protrusion rate that is difficult to treat with transcervical resection.


Assuntos
Leiomioma , Embolização da Artéria Uterina , Neoplasias Uterinas , Gravidez , Feminino , Humanos , Neoplasias Uterinas/terapia , Estudos Retrospectivos , Resultado do Tratamento , Leiomioma/diagnóstico por imagem , Leiomioma/terapia , Hormônios
3.
J Equine Vet Sci ; 133: 104990, 2024 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-38159581

RESUMO

Infectious ulcerative keratitis is a common disease in racehorses. To improve treatment outcomes, this study aimed to assess the antimicrobial susceptibilities of bacterial and fungal isolates obtained from the cornea of Japanese Thoroughbred racehorses with equine infectious ulcerative keratitis. Bacterial and fungal cultures were performed for 166 corneal swabs from 107 cases. A disc diffusion test and minimum inhibitory concentration test were also performed to assess antimicrobial susceptibility of the bacterial and fungal isolates, respectively. Bacterial and/or fungal isolates were obtained from 85.0% (91/107) of the cases. Staphylococcus was primarily isolated from bacterial isolates, including methicillin-resistant Staphylococcus aureus (MRSA), Aerococcus, Streptococcus, Acinetobacter, and Pseudomonas. Aspergillus was primarily isolated from filamentous fungi, and Debaryomyces species was primarily identified in yeast-like fungi. Ofloxacin resistance was observed in 100% (12/12), 15.9% (7/44), and 25.0% (3/12) of MRSA, Staphylococcus, and Streptococcus isolates, respectively. The prevalence of quinolone-resistant Staphylococci and Streptococci has increased in the past two decades. All Aspergillus isolates were susceptible to voriconazole, whereas other filamentous fungi, including Fusarium, were less susceptible to voriconazole. Further studies are required to determine effective treatments for antimicrobial-resistant isolates.


Assuntos
Anti-Infecciosos , Úlcera da Córnea , Doenças dos Cavalos , Ceratite , Staphylococcus aureus Resistente à Meticilina , Cavalos , Animais , Úlcera da Córnea/tratamento farmacológico , Úlcera da Córnea/epidemiologia , Úlcera da Córnea/veterinária , Voriconazol/farmacologia , Ceratite/tratamento farmacológico , Ceratite/epidemiologia , Ceratite/veterinária , Bactérias , Anti-Infecciosos/farmacologia , Doenças dos Cavalos/tratamento farmacológico , Doenças dos Cavalos/epidemiologia , Doenças dos Cavalos/microbiologia
4.
Int J Gynecol Pathol ; 2023 Nov 13.
Artigo em Inglês | MEDLINE | ID: mdl-38085954

RESUMO

Mesonephric-like adenocarcinoma (MLA) has recently been described as a tumor of the endometrium or ovaries, which, morphologically and immunohistochemically, resembles mesonephric adenocarcinoma arising mostly in the uterine cervix. Herein, we report, to our knowledge, the first case of ovarian MLA that developed into an extremely rapidly growing recurrent mesonephric-like carcinosarcoma, as confirmed by a genomic profiling test. A 51-year-old woman underwent chemotherapy with complete debulking surgery for ovarian carcinoma. Pathologically, the patient was diagnosed with stage IVB ovarian MLA. Subsequent to 15 months of complete remission, an enhanced computed tomography scan revealed a solid tumor of 10 cm diameter in the abdominal cavity. Secondary surgery was terminated with a 2 cm2 tumor biopsy specimen collection considering perioperative complications. Histologically, the tumor consisted of short spindle cells, and immunohistochemical staining revealed a rhabdomyosarcomatous profile without an epithelial component. Despite treatment for the sarcoma, she died 3 months after the detection of the tumor. The genomic profiling of the primary ovarian carcinoma and secondary resected tumor biopsy specimens revealed an identical KRAS mutation in both. Therefore, we concluded that the ovarian MLA recurred with a rhabdomyosarcoma component.

5.
J Equine Vet Sci ; 131: 104956, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37879453

RESUMO

Equine influenza virus (EIV) infection is one of the most important respiratory diseases in the equine industry around the world. Rapid diagnosis, facilitated by point-of-care testing, is essential to implement movement restrictions and control disease outbreaks. This study evaluated a microfluidic immunofluorescence assay kit, which detects influenza virus and SARS-CoV-2 antigens in human specimens with a 12 min turnaround time, for its potential use in detecting EIV. The microfluidic immunofluorescence assay kit succeeded in detecting 11 EIV strains. Using the real-time reverse transcription polymerase chain reaction as a reference assay, the microfluidic immunofluorescence assay kit showed a sensitivity of 60.7% when evaluating nasopharyngeal swab samples of three horses experimentally infected with EIV. Comparing with the other two rapid antigen detection kits based on immunochromatography and silver amplification immunochromatography, the microfluidic immunofluorescence assay kit exhibited higher sensitivity than the former assay (53.6%) and the same sensitivity as the latter (60.7%). The microfluidic immunofluorescence assay kit did not detect nine non-EIV viruses including one equine coronavirus strain and seven bacteria, suggesting a high specificity for EIV antigens. Similar to other rapid antigen detection kits, the microfluidic immunofluorescence assay kit could be an effective diagnostic tool to detect EIV in the field.


Assuntos
Doenças dos Cavalos , Vírus da Influenza A Subtipo H3N8 , Infecções por Orthomyxoviridae , Humanos , Animais , Cavalos , Infecções por Orthomyxoviridae/diagnóstico , Infecções por Orthomyxoviridae/veterinária , Microfluídica , Doenças dos Cavalos/diagnóstico , Imunofluorescência/veterinária
6.
Sci Rep ; 13(1): 13099, 2023 08 11.
Artigo em Inglês | MEDLINE | ID: mdl-37567893

RESUMO

We encountered 34 Clostridioides difficile (C. difficile) infection (CDI) cases among Thoroughbred horses in Japan from 2010 to 2021. Among them, 79.4% (27/34) either died or were euthanised. The risk factors associated with CDI and mortality among Japanese Thoroughbred horses remain unclear. We used genetic methods to examine C. difficile strains and their relationships with prognosis. Twenty-two (64.7%) cases were hospitalised at the onset of colitis. Outcomes were balanced for hospitalisation rates at the onset of colitis. The mortality rates of cases treated with metronidazole (65.0%) were significantly lower than untreated cases (100%). The predominant genotype of C. difficile isolate was polymerase chain reaction ribotype (RT) 078, isolated from 12 cases (35.3%), followed by RT014 (six cases, 17.6%). Binary toxin (C. difficile transferase [CDT])-positive strains, including all RT078 strains, were isolated from 16 horses. Mortality rates in RT078 strain (75.0%) or CDT-positive strain (83.3%) cases were comparable to that in cases of other types. Sufficient infection control is needed to prevent CDI in Thoroughbred horses. A timely and prompt CDI diagnosis leading to metronidazole treatment would improve CDI outcomes.


Assuntos
Clostridioides difficile , Infecções por Clostridium , Cavalos/genética , Animais , Metronidazol/uso terapêutico , Clostridioides difficile/genética , Japão/epidemiologia , Infecções por Clostridium/tratamento farmacológico , Infecções por Clostridium/epidemiologia , Infecções por Clostridium/veterinária , Ribotipagem
7.
J Vet Med Sci ; 85(7): 751-754, 2023 Jul 17.
Artigo em Inglês | MEDLINE | ID: mdl-37258221

RESUMO

A two-dose revaccination against tetanus is recommended for horses over 2 years old in Japan with no history of vaccination in the previous year. Here, the need for two-dose revaccination was evaluated in terms of antibody titers for each vaccine type, namely monovalent or multivalent. There was no difference in antibody titers between one- and two-dose regimens for up to 1 year, except at 8 weeks with the multivalent vaccine, and all horses had sufficient antibody titers for 1 year of tetanus prophylaxis. These results suggest that one-dose revaccination, regardless of the vaccine type, is as effective as two-dose in preventing tetanus for at least 1 year in horses not vaccinated in the previous year.


Assuntos
Doenças dos Cavalos , Tétano , Cavalos , Animais , Tétano/prevenção & controle , Tétano/veterinária , Imunização Secundária/veterinária , Toxoide Tetânico , Vacinação/veterinária , Japão , Anticorpos Antibacterianos , Doenças dos Cavalos/prevenção & controle
8.
Arch Virol ; 168(2): 35, 2023 Jan 07.
Artigo em Inglês | MEDLINE | ID: mdl-36609628

RESUMO

Mosquitoes and EDTA-treated blood samples from febrile racehorses were investigated for Getah virus infection from 2016 to 2019 at the Miho Training Center, where several outbreaks of Getah virus have occurred. We collected 5557 mosquitoes and 331 blood samples from febrile horses in this study. The most frequently captured mosquito species was Culex tritaeniorhynchus (51.9%), followed by Aedes vexans nipponii (14.2%) and Anopheles sinensis (11.2%). Getah virus was detected in mosquitoes (Aedes vexans nipponii) in 2016 (strain 16-0810-26) but not in 2017-2019. Six of 74 febrile horses in 2016 and one of 69 in 2019 tested positive for Getah virus; none of the horses tested positive in 2017 or 2018. Phylogenetic and sequence analysis showed that strain 16-0810-26 was closely related to strains that had been isolated from horses and a pig around the training center in 2014-2016 but have not been detected in samples collected at the training center since 2017. In contrast, the strain isolated from the infected horse in 2019 (19-I-703) was genetically distinct from the strains isolated from horses and a pig in 2014-2016 and was more closely related to a strain isolated in 1978 at the training center. The source of strain 19-I-703 is unclear, but the virus was not detected in other horses sampled in 2019. In summary, we found that the distribution of mosquito species present at the training center had not changed significantly since 1979, and although a small outbreak of Getah virus infection occurred among horses at the training center in 2016, limited Getah virus activity was detected in mosquitoes and horses at the training center from 2017 to 2019.


Assuntos
Aedes , Alphavirus , Viroses , Cavalos , Animais , Suínos , Japão/epidemiologia , Filogenia , Surtos de Doenças/veterinária , Viroses/epidemiologia
9.
Arch Biochem Biophys ; 729: 109379, 2022 10 30.
Artigo em Inglês | MEDLINE | ID: mdl-36002083

RESUMO

Obesity is a risk factor for gastrointestinal malignancies and tumors. However, which factors either protect or predispose the gastrointestinal organs to high-fat diet (HFD)-induced neoplasia remains unclear. Here, we demonstrate that HFD impacts the stomach to a greater extent as compared to the colorectum, resulting in leptin receptor (LepR) signaling-mediated neoplasia in the tissues. HFD activated leptin signaling, which in turn, accelerates the pathogenesis in the gastric mucosa more than that in the colorectum along with ectopic TFF3 expression. Moreover, in the stomach, higher levels of phosphorylated epidermal growth factor receptor (EGFR) in addition to the activation of STAT3 and Akt were observed as compared to the colorectum. The mice with LepR deletion in the gastrointestinal epithelium exhibited a suppressed induction of leptin, TFF3, and phosphorylated EGFR in the stomach, whereas the levels in the colorectum were insignificant. In co-transfected COS-7 cells with LepR and EGFR plasmid DNA, leptin transactivated EGFR to accelerate TFF3 induction along with activation of STAT3, ERK1/2, Akt, and PI3K p85/p55. Furthermore, TFF3 could bind to EGFR but did not transactivate LepR. Leptin-induced TFF3 induction was markedly suppressed by inhibitors of PI3K (LY294002) and EGFR (Erlotinib). Together, these results suggest a novel role of LepR-mediated signaling in transactivating EGFR that leads to TFF3 expression via the PI3K-Akt pathway. Therefore, this study sheds light on the identification of potentially new therapeutic targets for the treatment of pre-cancerous symptoms in stomach and colorectum.


Assuntos
Leptina , Receptores para Leptina , Animais , DNA , Gorduras na Dieta/efeitos adversos , Receptores ErbB/metabolismo , Cloridrato de Erlotinib , Leptina/metabolismo , Camundongos , Camundongos Obesos , Fosfatidilinositol 3-Quinases/metabolismo , Proteínas Proto-Oncogênicas c-akt/metabolismo , Receptores para Leptina/genética , Receptores para Leptina/metabolismo , Estômago/patologia , Ativação Transcricional , Fator Trefoil-3/genética , Fator Trefoil-3/metabolismo
10.
Vet Anim Sci ; 17: 100259, 2022 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-35800153

RESUMO

Methicillin-resistant Staphylococcus aureus (MRSA) has often been isolated from livestock and companion animals, including horses. Seven cases of MRSA infection in Thoroughbred racehorses were observed in an equine hospital in Japan in 2020. In this study, MRSA isolates from these seven horses and nine veterinarians in the equine hospital were studied to examine their genetic relatedness and evaluate the possibility of MRSA transmission. The MRSA isolates were subjected to whole-genome sequencing for multi-locus sequence typing, S. aureus protein A (spa) typing, staphylococcal cassette chromosome typing, and antimicrobial resistance gene detection. Minimum inhibitory concentrations of antibiotics were assessed to determine the antimicrobial susceptibility phenotype of the isolates. Phylogenetic trees based on single nucleotide polymorphisms were constructed to identify genetically close isolates. All isolates from horses and veterinarians belonged to sequence type (ST) 1, spa type t1784, with a point mutation in gyrA and double point mutations in grlA, which is known to cause fluoroquinolone resistance. All ST1-t1784 isolates were genetically closely related based on the phylogenetic tree. Our results suggested an outbreak and horse-veterinarian transmission of ST1-t1784 strains in an equine hospital.

11.
J Equine Vet Sci ; 114: 104004, 2022 07.
Artigo em Inglês | MEDLINE | ID: mdl-35526726

RESUMO

Antimicrobial administration can lead to imbalances of gastrointestinal microbiota, called dysbiosis. Dysbiosis sometimes results in diarrhea and enteritis in horses. Fecal microbiota transplantation (FMT) is used to treat affected horses, but whether it is effective as a prophylactic approach for dysbiosis in horses receiving antimicrobials remains unknown. The aim of this study was to assess the efficacy of simultaneous FMT against metronidazole-induced dysbiosis in horses. Changes in the ratios of bacterial families, determined by metagenomic analysis, were similar between the metronidazole-treated group and the simultaneous metronidazole- and FMT-treated group, notably in the Clostridiaceae, Ruminococcaceae, and Enterobacteriaceae. Differences in fecal bacterial compositions were due mainly to metronidazole administration (P = .0003), but not to FMT (P = .3136). Simultaneous FMT at 500 g of donor feces in 1 L of suspension once a day did not inhibit metronidazole-induced dysbiosis. The results show that the FMT protocol needs to be improved to prevent metronidazole-induced gut dysbiosis in horses.


Assuntos
Disbiose , Microbioma Gastrointestinal , Doenças dos Cavalos , Animais , Disbiose/induzido quimicamente , Disbiose/veterinária , Transplante de Microbiota Fecal/métodos , Transplante de Microbiota Fecal/veterinária , Fezes/microbiologia , Doenças dos Cavalos/induzido quimicamente , Cavalos , Metronidazol
12.
Arch Virol ; 167(8): 1611-1618, 2022 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-35639190

RESUMO

Equine coronavirus (ECoV) causes pyrexia, anorexia, lethargy, and sometimes diarrhoea. Infected horses excrete the virus in their faeces, and ECoV is also detected in nasal samples from febrile horses. However, details about ECoV infection sites in the intestinal and respiratory tracts are lacking. To identify the ECoV infection sites in the intestinal and respiratory tracts, we performed an experimental infection study and analysed intestinal and respiratory samples collected from four infected horses at 3, 5, 7, and 14 days post-inoculation (dpi) by real-time reverse transcription polymerase chain reaction (real-time RT-PCR) and in situ hybridization (ISH). Two horses became febrile, but the other two did not. None of the horses had diarrhoea or respiratory signs, and severe cases were not observed in this study. None of the horses showed obvious abnormalities in their intestinal or respiratory tracts. Real-time RT-PCR and ISH showed that ECoV RNA was present throughout the intestinal tract, and ECoV-positive cells were mainly detected on the surface of the intestine. In one horse showing viremia at 3 dpi, ECoV RNA was detected in the lung by real-time RT-PCR, but not by ISH. This suggests that the lung cells themselves were not infected with ECoV and that real-time RT-PCR detected viremia in the lung. The other three horses were positive for ECoV RNA in nasal swabs but were negative in the trachea and lung by real-time RT-PCR and ISH. This study suggests that ECoV broadly infects the intestinal tract and is less likely to infect the respiratory tract.


Assuntos
Betacoronavirus 1 , Infecções por Coronavirus , Doenças dos Cavalos , Animais , Infecções por Coronavirus/veterinária , Diarreia , Febre , Cavalos , Intestinos , RNA , Sistema Respiratório , Viremia
13.
J Equine Sci ; 33(4): 71-74, 2022 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-36699202

RESUMO

Equine proliferative enteropathy (EPE) is an equine infectious disease that can lead to severe weight loss and hyperplasia of the intestinal mucosa due to infection with Lawsonia intracellularis. In this study, we investigated the prevalence of EPE in a major Thoroughbred breeding area: Hidaka district, Hokkaido, Japan. Of the 252 symptomatic horses that we tested, 192 EPE cases (76.2%), including 8 fatal cases, were confirmed from April 2015 to March 2020 by etiological and/or serological investigation. Most of the EPE cases were observed in foals (88.5%), with fewer cases in yearlings (7.3%) and adults (4.2%). Asymptomatic infection was observed in 62.9% of the horses kept with affected horses. These results suggest that EPE is an enzootic disease in Hidaka district.

14.
J Vet Med Sci ; 84(1): 129-132, 2022 Jan 24.
Artigo em Inglês | MEDLINE | ID: mdl-34853198

RESUMO

Taylorella equigenitalis causes contagious equine metritis. Here we compared seven nucleic acid amplification tests for T. equigenitalis to select a rapid and reliable diagnostic method. The 95% detection limits of each assay varied greatly: real-time PCR had the lowest detection limit (0.77 fg/reaction); those of some of the conventional PCRs (cPCRs) were >100 fg/reaction. In experimentally infected samples, real-time PCR and semi-nested PCR showed the highest positive numbers (33 out of 42 samples), but two of the cPCRs detected only 2 and 7 positive results. Our results indicate that the use of sensitive molecular assays is important for the efficient detection of T. equigenitalis in clinical samples.


Assuntos
Infecções por Bactérias Gram-Negativas , Doenças dos Cavalos , Taylorella equigenitalis , Animais , Infecções por Bactérias Gram-Negativas/diagnóstico , Infecções por Bactérias Gram-Negativas/veterinária , Doenças dos Cavalos/diagnóstico , Cavalos , Técnicas de Amplificação de Ácido Nucleico/veterinária , Reação em Cadeia da Polimerase em Tempo Real/veterinária , Taylorella equigenitalis/genética
15.
J Vet Med Sci ; 83(12): 1907-1912, 2021 Dec 09.
Artigo em Inglês | MEDLINE | ID: mdl-34732605

RESUMO

Mycoplasma species are often isolated from horses with respiratory symptoms; however, the pathogenicity of Mycoplasma is still unclear. In autumn of 2018, we encountered an increase in cases with respiratory symptoms, mainly coughing, in a group of Thoroughbred racehorses in Japan. We examined tracheal wash samples obtained from 40 of those cases. Bacteria and viruses that commonly cause respiratory symptoms were investigated, and anaerobes were detected in only 5 cases and Streptococcus equi subsp. zooepidemicus (S. zooepidemicus) was detected in only 1 case of 40 cases with loop-mediated isothermal amplification assay. S. zooepidemicus and Streptococcus pneumoniae were isolated at a bacterial count of higher than 1.0 × 104 CFU/ml from 5 and 2 cases of 28 cases cultured, respectively. None of the viruses investigated was detected in 40 cases. Mycoplasma equirhinis (M. equirhinis) was isolated from 40.0% (16/40) of the cases, which was higher than previously reported isolation rates. The rate of M. equirhinis isolation in the cases from 2018 was significantly higher than the isolation rates in the other horses: clinical cases with respiratory symptoms in 2019-2020 (13.6%, 3/22) and healthy horses (13.5%, 5/37) in Japan. In this study, the isolation rate of M. equirhinis from horse group with cough symptoms in 2018 was high and no other common etiological agents were detected. The pathogenesis of M. equirhinis is still unclear, however, M. equirhinis might have been associated with respiratory symptoms in the Thoroughbred horse cases in 2018.


Assuntos
Doenças dos Cavalos , Mycoplasma , Infecções Estreptocócicas , Streptococcus equi , Animais , Doenças dos Cavalos/epidemiologia , Cavalos , Prevalência , Infecções Estreptocócicas/epidemiologia , Infecções Estreptocócicas/veterinária
16.
J Glob Antimicrob Resist ; 27: 225-227, 2021 12.
Artigo em Inglês | MEDLINE | ID: mdl-34655811

RESUMO

OBJECTIVES: Extended-spectrum ß-lactamase (ESBL)-producing Enterobacteriaceae have become a cause for great concern. Although some studies have reported the prevalence of ESBL-producing bacteria and ESBL-encoding genes in horses worldwide, the genetic structure surrounding the ESBL gene has not been analysed in detail. In the present study, we isolated two ESBL-producing Escherichia coli strains from diseased racehorses in Japan and demonstrated the mechanisms underlying the acquisition of their antimicrobial resistance (AMR) genes. METHODS: Two ESBL-producing E. coli strains (E148 and E189) were isolated from the heart and liver of horses with endocarditis and sepsis in 2014 and 2016, respectively, in Japan. Complete genomic sequences of the two strains were analysed using a PacBio RSII sequencer. Antimicrobial susceptibility testing was performed by the agar dilution method. RESULTS: The two isolates possessed a chromosomal AMR gene cluster containing blaCTX-M-1 that was similar to the pEQ1 plasmid found in E. coli isolated from a racehorse in the Czech Republic. In one of the two strains, tandem duplication of the 16-kb region containing blaCTX-M-1 and a class 1 integron, which occurred via IS26-mediated recombination, increased minimum inhibitory concentrations (MICs) associated with the duplicated AMR genes. CONCLUSION: Chromosomal blaCTX-M-1 possibly derived from the pEQ1 or pEQ1-like plasmid was found in Japanese equine E. coli isolates. In Japanese strains, many AMR genes containing blaCTX-M-1 and the class 1 integron are highly accumulated in one region on the chromosome, and the AMR of E. coli was enhanced via the IS26-mediated duplication of the AMR gene cluster.


Assuntos
Anti-Infecciosos , Escherichia coli , Animais , Antibacterianos/farmacologia , Cromossomos , Farmacorresistência Bacteriana , Escherichia coli/genética , Cavalos , Integrons , Japão , beta-Lactamases/genética
17.
J Equine Vet Sci ; 103: 103664, 2021 08.
Artigo em Inglês | MEDLINE | ID: mdl-34281642

RESUMO

Matrix-assisted laser desorption ionization-time of flight mass spectrometry (MALDI-TOF MS) is used for identification of bacterial species isolated from horses. However, because of insufficiencies in the reference database, some bacterial species isolated from horses are difficult to identify with MALDI-TOF MS, and enriching the databases is expected to enhance the accuracy of MALDI-TOF MS identification. Here we created an in-house database including 271 bacterial isolates from horses. Furthermore, we used an enhanced database (our in-house database plus a commercially provided database) to examine 91 newly obtained isolates that could not be identified with MALDI-TOF MS using the commercially provided database. The enhanced database could identify 15 of those 91 isolates to the species level; including streptococcus (3/19), Gram-positive rod (4/17), Gram-negative rod (8/17) isolates. The enhanced database increased the average identification score of the 91 isolates (1.64-1.76). The in-house database increased the number of isolates that MALDI-TOF MS could identify to the species level and contributed to more accurate identification of bacterial isolates from horses.


Assuntos
Bactérias , Animais , Bases de Dados Factuais , Cavalos , Espectrometria de Massas por Ionização e Dessorção a Laser Assistida por Matriz/veterinária
18.
Sci Rep ; 11(1): 11884, 2021 06 04.
Artigo em Inglês | MEDLINE | ID: mdl-34088956

RESUMO

Microbial communities are commonly studied by using amplicon sequencing of part of the 16S rRNA gene. Sequencing of the full-length 16S rRNA gene can provide higher taxonomic resolution and accuracy. To obtain even higher taxonomic resolution, with as few false-positives as possible, we assessed a method using long amplicon sequencing targeting the rRNA operon combined with a CCMetagen pipeline. Taxonomic assignment had > 90% accuracy at the species level in a mock sample and at the family level in equine fecal samples, generating similar taxonomic composition as shotgun sequencing. The rRNA operon amplicon sequencing of equine fecal samples underestimated compositional percentages of bacterial strains containing unlinked rRNA genes by a fourth to a third, but unlinked rRNA genes had a limited effect on the overall results. The rRNA operon amplicon sequencing with the A519F + U2428R primer set was able to detect some kind of archaeal genomes such as Methanobacteriales and Methanomicrobiales, whereas full-length 16S rRNA with 27F + 1492R could not. Therefore, we conclude that amplicon sequencing targeting the rRNA operon captures more detailed variations of equine microbiota.


Assuntos
DNA Intergênico , Microbioma Gastrointestinal , Microbiota , RNA Ribossômico 16S/genética , RNA Ribossômico 23S/genética , Análise de Sequência de DNA/métodos , Animais , Bactérias/genética , DNA Bacteriano/genética , Fezes , Feminino , Sequenciamento de Nucleotídeos em Larga Escala/métodos , Cavalos , Filogenia , Análise de Sequência de DNA/instrumentação , Óperon de RNAr
19.
Microb Genom ; 7(3)2021 03.
Artigo em Inglês | MEDLINE | ID: mdl-33684029

RESUMO

The equine disease strangles, which is characterized by the formation of abscesses in the lymph nodes of the head and neck, is one of the most frequently diagnosed infectious diseases of horses around the world. The causal agent, Streptococcus equi subspecies equi, establishes a persistent infection in approximately 10 % of animals that recover from the acute disease. Such 'carrier' animals appear healthy and are rarely identified during routine veterinary examinations pre-purchase or transit, but can transmit S. equi to naïve animals initiating new episodes of disease. Here, we report the analysis and visualization of phylogenomic and epidemiological data for 670 isolates of S. equi recovered from 19 different countries using a new core-genome multilocus sequence typing (cgMLST) web bioresource. Genetic relationships among all 670 S. equi isolates were determined at high resolution, revealing national and international transmission events that drive this endemic disease in horse populations throughout the world. Our data argue for the recognition of the international importance of strangles by the Office International des Épizooties to highlight the health, welfare and economic cost of this disease. The Pathogenwatch cgMLST web bioresource described herein is available for tailored genomic analysis of populations of S. equi and its close relative S. equi subspecies zooepidemicus that are recovered from horses and other animals, including humans, throughout the world. This article contains data hosted by Microreact.


Assuntos
Doenças dos Cavalos/microbiologia , Doenças dos Cavalos/transmissão , Infecções Estreptocócicas/veterinária , Streptococcus equi/isolamento & purificação , Animais , Feminino , Genoma Bacteriano , Cavalos , Masculino , Filogenia , Infecções Estreptocócicas/microbiologia , Infecções Estreptocócicas/transmissão , Streptococcus equi/classificação , Streptococcus equi/genética , Streptococcus equi/fisiologia
20.
Vet Med Sci ; 7(3): 621-625, 2021 05.
Artigo em Inglês | MEDLINE | ID: mdl-33336899

RESUMO

Bacterial placentitis in horses commonly results in abortion, premature birth or compromised neonatal foal health. Although mycobacterial infections are generally uncommon in horses, 10 equine abortion cases caused by Mycobacterium avium subsp. hominissuis (MAH) infections occurred between 2018 and 2019 in Japan. They occurred on seven Thoroughbred horse farms in the Hidaka district of Hokkaido, but direct contact among the mares on different farms was not recorded. Most cases were characterized by extensive pathological lesions of the placenta, which are not typical in cases of common pathogenic bacteria such as Streptococcus zooepidemicus and Escherichia coli. All abortions featured white-yellow exudates on the surface of the placenta. Mycobacterial granuloma formations were histologically found in the placenta and fetal organs, and acid-fast bacteria were isolated from the placenta, fetal samples (heart, lung, liver, kidney, spleen and stomach contents) or uterine lavage fluid. The greatest number of bacteria was isolated from necrotic lesions on the placenta, which could be an important site for bacterial isolation in mycobacterial equine abortions. The isolates were identified as MAH based on internal genome sequences. In variable number tandem repeat analysis, all patterns of the strains were identical. Single nucleotide polymorphism analysis of the core genome grouped all strains in the II-a/SC3 subcluster. Both results reveal that these strains share the same genetic background, suggesting that the horses had been infected by the same unknown contagious source.


Assuntos
Aborto Animal/microbiologia , Doenças dos Cavalos/microbiologia , Infecções por Mycobacterium/veterinária , Mycobacterium/fisiologia , Animais , Cavalos , Japão , Infecções por Mycobacterium/microbiologia
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