Efficacy of antimicrobial therapy for bovine acute Klebsiella pneumoniae mastitis.

The purpose of this retrospective study was to assess the efficacy of antimicrobial therapy for bovine acute Klebsiella pneumoniae mastitis. We evaluated data from cattle in Ehime, Japan, with naturally occurring acute mastitis due to K. pneumoniae (n=208) or Escherichia coli (n=201). Survival was significantly shorter in cattle with acute K. pneumoniae mastitis (median, 76 days) compared with the disease caused by E. coli (median 464 days). In 2004-2008, because both species were highly susceptible to cefazolin, cases of K. pneumoniae and E. coli mastitis were treated solely with cefazolin, yielding clinical cure rates of 52.8% for K. pneumoniae and 86.0% for E. coli. However, since 2009, the efficacy of treatment of K. pneumoniae mastitis with cefazolin alone has decreased. When cefazolin administered on the first disease day led to clinical improvement, treatment with cefazolin was continued. However, when cefazolin administered on the first disease day failed to yield clinical improvement, the antibiotic was switched to a fluoroquinolone on the second day, resulting in cure rates of 76.7% for K. pneumoniae and 80.0% for E. coli. These findings suggest that, when the first-line drug (e.g., cefazolin) is ineffective, promptly changing to a second-line drug (e.g., a fluoroquinolone) increases the cure rate for bovine K. pneumoniae mastitis.

Severe fishhook-related ocular injury: A case series.

This study examined the treatment, clinical course, and prognosis of two patients who visited our institution with severe ocular fishhook-related injuries with complications. The first patient was a 57-year-old man injured in the right eye by a fishhook lacerating the right upper eyelid. Although no aqueous humor leakage was observed, intraocular hemorrhage was severe, and the best-corrected visual acuity (BCVA) was "counting fingers" at 15 cm. The eyelid was sutured and vitreous surgery with cataract surgery was performed for traumatic cataract, vitreous hemorrhage, and retinal detachment. The patient experienced recurring iritis and the BCVA recovered to 20/100 eight months postoperatively. The second patient was a 62-year-old man who incurred a penetrating right-eye fishhook injury on a ship with BCVA of "hand motion." The sclera and iris were ruptured with severe hemorrhage and a shallow anterior chamber without leakage of aqueous humor. A damaged lens and vitreous hemorrhage were observed with intraocular pressure of 38 mmHg. The ruptured sclera was sutured and vitreous surgery with lensectomy was performed. After 16 months, the BCVA improved to 20/40; however, glaucoma control was maintained by topical medication. Therefore, ocular fishhook-related injury could result in irreversible visual impairment. It is important to pay attention and protect the eyes from moving fishhooks during fishing. PLAIN LANGUAGE SUMMARY: There are few reports on fishhook injury of the eye with resulting complications such as retinal detachment. We present the characteristics of the injury, treatment, clinical course, and prognosis of two patients with severe fishhook-related injuries of the eye with complications. Following treatment, most complications, including vitreous hemorrhage, detached lens, and retinal detachment, safely resolved in these cases; however, both patients required further treatment for recurring inflammation of the iris or glaucoma. The visual acuity of both patients improved over several months. Fishhook-related injuries of the eyes might result in irreversible visual impairment. It is important to pay attention and protect the eyes from moving fishhooks during fishing.

Epidemiological Study of Pathogenic Leptospira in Raccoons (Procyon lotor) in a Suburb of Tokyo, Japan.

Leptospirosis is a zoonosis that affects humans and animals worldwide. Raccoons (Procyon lotor), adopted in urban environments, may act as potential reservoirs of Leptospira. We investigated the prevalence of pathogenic Leptospira in the kidney and urine samples of raccoons living in Tokyo, as well as anti-leptospiral antibodies in their serum, and aimed to examine the factors that expose raccoons to Leptospira. Polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA) were used to detect leptospiral DNA and anti-leptospiral antibodies, respectively. Thirty-six of 156 raccoons (23.1%) were positive by PCR, and 16 of 165 raccoons (9.7%) were positive by ELISA. The prevalence and seroprevalence rates differed depending on the raccoon dispersal period. We used univariable logistic regression to estimate the environmental factors associated with pathogenic Leptospira and anti-leptospiral antibodies in raccoons. Significant differences were observed in the PCR results for the seasons (spring−summer) (p = 0.01), average monthly temperature (p < 0.01), and average monthly rainfall (p < 0.01). No significant difference was seen in the ELISA results, but raccoons in larger urban areas tended to have higher seroprevalence rates (p = 0.06). We identified a pattern of leptospiral spread in raccoon dispersal and environmental factors that expose raccoons to Leptospira.

Fecal microbiota transplantation as a new treatment for canine inflammatory bowel disease.

In human medicine, fecal microbiota transplantation (FMT) is an effective treatment for recurrent Clostridioides difficile infection. It has also been tested as a treatment for multiple gastrointestinal diseases, including inflammatory bowel disease (IBD). However, only a few studies have focused on the changes in the microbiome following FMT for canine IBD. Here, we performed FMT in nine dogs with IBD using the fecal matter of healthy dogs and investigated the subsequent changes in the fecal microbiome and clinical signs. In three dogs, the fecal microbiome was examined by 16S rRNA sequencing. Fusobacteria were observed at a low proportion in dogs with IBD. However, the post-FMT microbiome became diverse and showed a significant increase in Fusobacteria proportion. Fusobacterium was detected in the nine dogs by quantitative polymerase chain reaction. The proportion of Fusobacterium in the post-FMT fecal microbiome was significantly increased (p<0.05). The changes in clinical signs (e.g., vomiting, diarrhea, and weight loss) were evaluated according to the canine inflammatory bowel disease activity index. The score of this index significantly decreased in all dogs (p<0.05) with improvements in clinical signs. These improvements were related to the changes in the proportion of microbes, particularly the increase in Fusobacterium. The dogs with IBD showed a lower proportion of Fusobacterium than healthy dogs. This suggests that a low proportion of Fusobacterium is a characteristic feature of canine IBD and that Fusobacterium is involved in this disease. The results of this study may help elucidate the pathogenesis of this disease and its association with Fusobacterium.

Assessment of the Usefulness of Cefapirin and Cefalonium Disks for Susceptibility Testing of Staphylococcus aureus Isolates from Bovine Mastitis.

Cefapirin (CEP) and cefalonium (CNM) are first-generation cephalosporins widely used to treat bovine mastitis caused by Gram-positive bacteria including staphylococci. However, disks for susceptibility testing of those drugs in causative bacteria are not available. This study evaluated the efficacy of 10 µg and 30 µg pilot disks of CEP (CEP10 and CEP30) and CNM (CNM10 and CNM30) against 130 Staphylococcus aureus isolates from bovine mastitis. Scattergrams of minimum inhibitory concentrations (MICs) and zone diameters (ZDs) illustrated significant correlations between the MICs and ZDs of CEP10 (r = -0.912), CEP30 (r = -0.933), CNM10 (r = -0.847), and CNM30 (r = -0.807). The analysis by Normalized Resistance Interpretation indicated that the epidemiolocal cut-off value (ECV) of MIC for both cefapirin and cefalonium is ≤ 0.5 µg/mL, and the ECV of ZD for CEP10, CEP30, CNM10, and CNM30 were ≥ 22 mm, ≥ 25 mm, ≥ 22 mm, and ≥ 29 mm, respectively. We believe that both 10 µg and 30 µg CEP and CNM susceptibility disks will be helpful for guiding the appropriate use of these antibiotics for bovine mastitis. Further studies toward the establishment of clinical breakpoint of CEP and CNM would be needed for their routine use.

Improvement in Clinical Symptoms and Fecal Microbiome After Fecal Microbiota Transplantation in a Dog with Inflammatory Bowel Disease.

PURPOSE: Recently, fecal microbiota transplantation (FMT) has been tested in veterinary medicine as a treatment option for multiple gastrointestinal (GI) diseases, such as inflammatory bowel disease (IBD). However, there are no reports of changes in the microbial diversity of fecal microbiome after treatment with FMT in canine IBD cases. Moreover, little is known about the long-term efficacy and safety of FMT treatment for dogs. Herein, we present a case of canine intractable IBD treated with repeated, long-term FMT. PATIENTS AND METHODS: The patient was a 10-year-old, neutered, male, 4-kg Toy Poodle with a prolonged history of vomiting and diarrhea. Fecal examination for pathogens was negative. Despite treatment with multiple antibacterial and antidiarrheal agents, the patient showed no improvement. Endoscopic mucus sampling diagnosed a case of lymphocytic-plasmacytic duodenitis, ie, idiopathic IBD. Eventually, we performed periodic, long-term fecal microbiota transplantation of fresh donor feces collected from a 4-year-old, 32.8-kg, neutered male Golden Retriever by rectal enema. Additionally, we performed 16S rRNA sequence analysis, before and after FMT, to evaluate the microbiome diversity. RESULTS: Fecal microbiome diversity after FMT resembled that of the healthy donor dog's fecal microbiome, before FMT, which led us to conclude that the fecal microbiome in our patient normalized with FMT. Moreover, the clinical symptoms improved remarkably with regard to the changes in the fecal microbiome. Additionally, we noted no observable side effects during FMT treatment. CONCLUSION: This report indicates the efficacy and safety of long-term, periodic FMT for a case of canine IBD based on attenuation of clinical symptoms and changes in fecal microbiome diversity. Therefore, FMT could be chosen as a treatment option for IBD in canines in the future.

Comparison of the effects of four commercially available prescription diet regimens on the fecal microbiome in healthy dogs.

The effects of prescription diets on canine intestinal microbiota are unknown. In this study, we used next generation sequencing to investigate the impact of four commercially available prescription diet regimens on the fecal microbiome in six healthy dogs. The diet regimens used were as follows: weight-loss diet, low-fat diet, renal diet, and anallergenic diet. We found a significantly decreased proportion of phylum Actinobacteria with the weight-loss diet compared to the anallergenic diet. There were no significant differences in the proportion of phylum Bacteroidetes between the four diets. The proportion of phylum Firmicutes was significantly decreased with the weight-loss diet compared to the anallergenic diet. The proportion of phylum Fusobacteria was significantly increased with the weight-loss diet compared to the anallergenic diet. There were no significant differences in the proportion of phylum Proteobacteria after consumption of the four diets. We therefore demonstrated that commercial prescription diet influences the fecal microbiome in healthy dogs. These results might be useful when choosing a prescription diet for targeting a disease.

Spontaneous resolution of myopic foveoschisis and a macular hole with retinal detachment.

PURPOSE: The aim of this report was to describe a case of myopic foveoschisis and a macular hole with retinal detachment that resolved spontaneously. OBSERVATIONS: A 62-year-old woman with bilateral blurred vision was referred to our department. Her best-corrected visual acuity (BCVA) was 0.4 bilaterally; the standard equivalent refractive error was -17.0 diopters in the right eye and -18.75 diopters in the left eye. The axial length was 31.4 mm and 31.2 mm, respectively. After cataract surgery was performed, the BCVA of the right and left eye was 0.6 and 1.0, respectively. Four years later, slight macular traction, foveoschisis, and the macular hole had progressed in the left eye and the BCVA had decreased to 0.7. However, the macular hole had closed, the foveoschisis had resolved, and retinal detachment had mostly resolved three weeks later. The BCVA improved to 0.8. The macular hole did not reoccur and retinal detachment resolved completely after six more months. CONCLUSIONS AND IMPORTANCE: This case demonstrated that myopic foveoschisis and a macular hole with retinal detachment could improve spontaneously. It is necessary to decide carefully whether surgery is required or whether the patient should be observed.

Comparison of two indirect ELISA coating antigens for the detection of dairy cow antibodies against Pasteurella multocida.

The ELISA is recognized as an efficient diagnostic tool for antibody detection, but there is no standard ELISA assay for detection of antibodies against hemorrhagic septicemia (HS) in cattle. The present study reports on an indirect ELISA assay for antibody detection of HS in dairy cows, and evaluates the sensitivity (Se) and specificity (Sp) of the method using a Bayesian approach. An indirect ELISA was developed with two types of heat extract antigens, Pasteurella multocida strains P-1256 and M-1404, as coating antigens. A checkerboard titration was employed using dairy cow sera immunized with P. multocida bacterin and colostrum-deprived calf sera. The concentrations of heat extract antigen (160µg/mL), sample serum (1:100) and goat anti-bovine immunoglobulin G labeled with horseradish peroxidase (1:2000) were optimal for the assay. The cut-off values were 0.147 and 0.128 for P-1256 and M-1404 coating antigens, and there were no differences in the results of tests with positive and negative sera (p<0.05). The characteristics of three diagnostic tests were evaluated using a one-population Bayesian model, assuming conditional dependence between two types of coating antigen-based ELISAs and indirect hemagglutination assay (IHA). A total of 415 sera samples from dairy cows without HS vaccination and no history of disease were tested. The Se and Sp of the P-1256 and M-1404 ELISAs were higher than those of the IHA. The Se and Sp of the P-1256 ELISA were 90.3% and 90.1%, while the Se and Sp of the M-1404 ELISA were 92.1% and 71.9%. The median values of Se and Sp from the IHA were 36.0% and 58.2%.

A prospective multicenter study on genome wide associations to ranibizumab treatment outcome for age-related macular degeneration.

We conducted a genome-wide association study (GWAS) on the outcome of anti-VEGF treatment for exudative age-related macular degeneration (AMD) in a prospective cohort. Four hundred and sixty-one treatment-naïve AMD patients were recruited at 13 clinical centers and all patients were treated with 3 monthly injections of ranibizumab followed by pro re nata regimen treatment for one year. Genomic DNA was collected from all patients for a 2-stage GWAS on achieving dry macula after the initial treatment, the requirement for an additional treatment, and visual acuity changes during the 12-month observation period. In addition, we evaluated 9 single-nucleotide polymorphisms (SNPs) in 8 previously reported AMD-related genes for their associations with treatment outcome. The discovery stage with 256 patients evaluated 8,480,849 SNPs, but no SNPs showed genome-wide level significance in association with treatment outcomes. Although SNPs with P-values of <5 × 10-6 were evaluated in replication samples of 205 patients, no SNP was significantly associated with treatment outcomes. Among AMD-susceptibility genes, rs10490924 in ARMS2/HTRA1 was significantly associated with additional treatment requirement in the discovery stage (P = 0.0023), and pooled analysis with the replication stage further confirmed this association (P = 0.0013). ARMS2/HTRA1 polymorphism might be able to predict the frequency of injection after initial ranibizumab treatment.

Development and standardization of an in-house indirect ELISA for detection of duck antibody to fowl cholera.

Serological tests, such as agglutination and indirect hemagglutination assay (IHA), have been used to identify antibodies against Pasteurella multocida in poultry sera, but none are highly sensitive. An enzyme-linked immunosorbent assays (ELISA) has been used with varying degrees of success in attempts to monitor seroconversion in vaccinated poultry, but are not suitable for diagnosis. Commercial ELISA kits are available for chickens and turkeys, but not for ducks. The present study reports development and standardization of an in-house indirect ELISA for detection of duck antibody to fowl cholera. The characteristics of ELISA and IHA were analyzed using a one population Bayesian model assuming conditional dependence between the two diagnostic tests. An in-house indirect ELISA was developed using a heat extract antigen of P. multocida strain X-73 as a coating antigen and horseradish peroxidase conjugated goat anti-duck IgG antibody (dIgG-HRP). The checkerboard titration method was done using sera from ducks immunized with P. multocida bacterin as positive sera and 1day old duckling sera as negative sera. The heat extract antigen at 1µg/ml, sample serum at a dilution of 1:100, and dIgG-HRP 1:2000 were optimal concentrations for the assay. The cut-off value was 0.200. Of the duck sera, 89.05% (244/274) were considered seropositive by ELISA. Estimates for sensitivity and specificity of ELISA were higher than prior values with medians of 94.7% [95% posterior probability interval (PPI)=89.6-98.2%] and 87.2% (PPI=68.2-98.3%). Estimates for sensitivity of IHA were lower than prior values (median=97.6, PPI=93.2-99.7%) while the specificity was close to the prior value (median=76.5, PPI=65.8-85.4%). This finding suggests that an in-house indirect ELISA can be used to detect duck antibody to fowl cholera.

Phenotypic and molecular characterization of antimicrobial resistance in Enterobacter spp. isolates from companion animals in Japan.

The emergence of antimicrobial resistance among Enterobacter spp., including resistance to extended-spectrum cephalosporins (ESC), is of great concern in both human and veterinary medicine. In this study, we investigated the prevalence of antimicrobial resistance among 60 isolates of Enterobacter spp., including E. cloacae (n = 44), E. aerogenes (n = 10), and E. asburiae (n = 6), from clinical specimens of dogs and cats from 15 prefectures in Japan. Furthermore, we characterized the resistance mechanisms harbored by these isolates, including extended-spectrum ß-lactamases (ESBLs) and plasmid-mediated quinolone resistance (PMQR); and assessed the genetic relatedness of ESC-resistant Enterobacter spp. strains by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Antimicrobial susceptibility testing demonstrated the resistance rates to ampicillin (93.3%), amoxicillin-clavulanic acid (93.3%), cefmetazole (93.3%), chloramphenicol (46.7%), ciprofloxacin (43.3%), tetracycline (40.0%), ceftazidime (33.3%), cefotaxime (33.3%), trimethoprim/sulfamethoxazole (28.3%), gentamicin (23.3%), and meropenem (0%). Phenotypic testing detected ESBLs in 16 of 18 ESC-resistant E. cloacae isolates but not in the other species. The most frequent ESBL was CTX-M-15 (n = 8), followed by SHV-12 (n = 7), and CTX-M-3 (n = 1). As for AmpC ß-lactamases, CMY-2 (n = 2) and DHA-1 (n = 2) were identified in ESC-resistant E. cloacae strains with or without ESBLs. All of the ESC-resistant E. cloacae strains also harbored one or two PMQRs, including qnrB (n = 15), aac(6')-Ib-cr (n = 8), and qnrS (n = 2). Based on MLST and PFGE analysis, E. cloacae clones of ST591-SHV-12, ST171-CTX-M-15, and ST121-CTX-M-15 were detected in one or several hospitals. These results suggested intra- and inter-hospital dissemination of E. cloacae clones co-harboring ESBLs and PMQRs among companion animals. This is the first report on the large-scale monitoring of antimicrobial-resistant isolates of Enterobacter spp. from companion animals in Japan.

Phenotypic and Molecular Characterization of Antimicrobial Resistance in Klebsiella spp. Isolates from Companion Animals in Japan: Clonal Dissemination of Multidrug-Resistant Extended-Spectrum ß-Lactamase-Producing Klebsiella pneumoniae.

The emergence of antimicrobial resistance in Klebsiella spp., including resistance to extended-spectrum cephalosporins (ESC) and fluoroquinolones, is of great concern in both human and veterinary medicine. In this study, we investigated the prevalence of antimicrobial resistance in a total of 103 Klebsiella spp. isolates, consisting of Klebsiella pneumoniae complex (KP, n = 89) and K. oxytoca (KO, n = 14) from clinical specimens of dogs and cats in Japan. Furthermore, we characterized the resistance mechanisms, including extended-spectrum ß-lactamase (ESBL), plasmid-mediated AmpC ß-lactamase (PABL), and plasmid-mediated quinolone resistance (PMQR); and assessed genetic relatedness of ESC-resistant Klebsiella spp. strains by multilocus sequence typing (MLST) and pulsed-field gel electrophoresis (PFGE). Antimicrobial susceptibility testing demonstrated that resistance rates to ampicillin, cephalothin, enrofloxacin, ciprofloxacin, trimethoprim/sulfamethoxazole, cefotaxime, gentamicin, tetracycline, chloramphenicol, amoxicillin-clavulanic acid, and cefmetazole were 98.1, 37.9, 37.9, 35.9, 35.0, 34.0, 31.1, 30.1, 28.2, 14.6, and 6.8%, respectively. Phenotypic testing detected ESBLs and/or AmpC ß-lactamases in 31 of 89 (34.8%) KP isolates, but not in KO isolates. Resistances to 5 of the 12 antimicrobials tested, as well as the three PMQRs [qnrB, qnrS, and aac(6')-Ib-cr], were detected significantly more frequently in ESBL-producing KP, than in non-ESBL-producing KP and KO. The most frequent ESBL was CTX-M-15 (n = 13), followed by CTX-M-14 (n = 7), CTX-M-55 (n = 6), SHV-2 (n = 5), CTX-M-2 (n = 2), and CTX-M-3 (n = 2). Based on the rpoB phylogeny, all ESBL-producing strains were identified as K. pneumoniae, except for one CTX-M-14-producing strain, which was identified as K. quasipneumoniae. All of AmpC ß-lactamase positive isolates (n = 6) harbored DHA-1, one of the PABLs. Based on MLST and PFGE analysis, ST15 KP clones producing CTX-M-2, CTX-M-15, CTX-M-55, and/or SHV-2, as well as KP clones of ST1844-CTX-M-55, ST655-CTX-M-14, and ST307-CTX-M-15, were detected in one or several hospitals. Surprisingly, specific clones were detected in different patients at an interval of many months. These results suggest that multidrug-resistant ESBL-producing KP were clonally disseminated among companion animals via not only direct but also indirect transmission. This is the first report on large-scale monitoring of antimicrobial-resistant Klebsiella spp. isolates from companion animals in Japan.

Serological surveillance for antibodies against Erysipelothrix species in wild boar and deer in Japan.

We investigated the seroprevalence of antibodies against Erysipelothrix in wild animals in Japan. Serum samples were collected from 48 wild boar, 26 Yezo deer and 26 Japanese deer in Japan. Growth agglutination (GA) test was performed to estimate antibody titers. As a result, positive results were obtained from 32 (66.7%), 1 (3.6%) and 6 (23.1%) samples from wild boar, Yezo deer and Japanese deer, respectively. Our findings suggest that wild animals may be an important reservoir of Erysipelothrix.

Area densitometry using rotating Scheimpflug photography for posterior capsule opacification and surface light scattering analyses.

PURPOSE: To compare area densitometry analysis using rotating Scheimpflug photography in quantifications of posterior capsule opacification (PCO) and surface light scattering with previous anterior-segment analyzer measurement. SETTING: Miyata Eye Hospital, Miyazaki, Japan. DESIGN: Prospective observational case series. METHODS: Scheimpflug images of eyes with foldable intraocular lenses (IOLs) were obtained using rotating and fixed Scheimpflug photography. Area densitometry on the posterior and anterior surfaces was conducted for PCO and surface light scattering analyses, respectively, with an identical area size. Correlation between two measurements was analyzed using linear regression. RESULTS: The study included 105 eyes of 74 patients who received IOLs 1 to 18 years (mean, 4.9 ± 4.5 years) postoperatively. In the PCO analysis on the posterior IOL surface, there was a significant correlation between the two measurements (P < .001, R(2) = 0.60). In the surface light scattering analysis, a significant and higher correlation was obtained (P < .001, R(2) = 0.91) until the fixed Scheimpflug photography exhibited saturation due to intensive scatterings. CONCLUSION: Area densitometry combined with a rotating Scheimpflug photography was exchangeable to previously established densitometry measurement, and allowed successive evaluation in longer-term observations.

Prospective Comparison of One-Piece and Three-Piece Tecnis Aspheric Intraocular Lenses: 1-year Stability and its Effect on Visual Function.

PURPOSE: To compare the stability and visual function of one-piece hydrophobic intraocular lens (IOL) (ZCB00, Abbot Medical Optics) and a three-piece hydrophobic IOL (ZA9003, Abbot Medical Optics) after cataract surgery. MATERIALS AND METHODS: Eighteen eyes of 18 patients with the one-piece IOL and 20 eyes of 20 patients with the three-piece IOL who underwent cataract surgery were evaluated prospectively on best corrected distance visual acuity, postoperative refractive error, IOL decentration and tilt, anterior chamber depth (ACD), spherical aberration and contrast sensitivity. Measurements were performed 1 week and 1, 3, 6 and 12 months postoperatively. The postoperative outcomes were compared between the two groups. RESULTS: In the one-piece group, a hyperopic refractive error (p < 0.002) and deeper ACDs during 1-year post-operatively were observed compared with the three-piece group. There were no significant differences in BCVA, IOL tilt, spherical aberration or contrast sensitivity. CONCLUSIONS: The stability of the one-piece IOL was greater than that of the three-piece IOL. The postoperative ACD and refractive error demonstrated that an anteriorly offset haptic in the one-piece IOL allowed incremental fixation to the posterior capsule.

Phenotypic and molecular characterization of antimicrobial resistance in Proteus mirabilis isolates from dogs.

Large-scale monitoring of resistance to 14 antimicrobial agents was performed using 103 Proteus mirabilis strains isolated from dogs in Japan. Resistant strains were analysed to identify their resistance mechanisms. Rates of resistance to chloramphenicol, streptomycin, enrofloxacin, trimethoprim/sulfamethoxazole, kanamycin, ampicillin, ciprofloxacin, cephalothin, gentamicin, cefoxitin and cefotaxime were 20.4, 15.5, 12.6, 10.7, 9.7, 8.7, 5.8, 2.9, 2.9, 1.9 and 1.9%, respectively. No resistance to ceftazidime, aztreonam or imipenem was found. Class 1 and 2 integrases were detected in 2.9 and 11.7% of isolates, respectively. Class 1 integrons contained aadB or aadB-catB-like-blaOXA10-aadA1, whereas those of class 2 contained sat-aadA1, dhfr1-sat-aadA1 or none of the anticipated resistance genes. Of five distinct plasmid-mediated quinolone-resistance (PMQR) genes, only qnrD gene was detected in 1.9% of isolates. Quinolone-resistance determining regions (QRDRs) of gyrA and parC from 13 enrofloxacin-intermediate and -resistant isolates were sequenced. Seven strains had double mutations and three had single mutations. Three of nine ampicillin-resistant isolates harboured AmpC-type ß-lactamases (i.e. blaCMY-2, blaCMY-4 and blaDHA-1). These results suggest that canine Proteus mirabilis deserves continued surveillance as an important reservoir of antimicrobial resistance determinants. This is the first report, to our knowledge, describing integrons, PMQRs and QRDR mutations in Proteus mirabilis isolates from companion animals.

Antimicrobial susceptibility of enterococcal species isolated from antibiotic-treated dogs and cats.

In this study, we examined the antimicrobial susceptibility of the enterococci isolated from dogs and cats in Japan during 2011-2012. Fecal samples were collected from 84 dogs and 16 cats that underwent antibiotic treatment. Enterococci were detected in 70 of 84 dogs (83.3%) and 7 of 16 cats (43.8%). The most prevalent Enterococcus species was Enterococcus faecalis (64.9%); Enterococcus faecium and Enterococcus durans were also isolated from 14 of 77 (18.2%) and 5 of 77 (6.5%) of these animals, respectively. The most active resistance was observed for erythromycin (44.2%) and oxytetracycline (44.2%), and there was considerable resistance to lincomycin (41.6%), gentamicin (31.2%) and kanamycin (31.2%). Compared with the results of a similar study conducted in 2006 and 2007, enterococci susceptibility to enrofloxacin and ampicillin had significantly increased. Enterococcus gallinarum harboring vanC1 and Enterococcus casseliflavus harboring vanC2/3 were isolated from 4 of 77 enterococcal isolates. However, no enterococcal isolates were resistant to vancomycin. Multidrug resistance was found for as few as two and as many as nine antimicrobials regardless of the class. These results demonstrate that dogs and cats treated with antibiotics are commonly colonized with antimicrobial-resistant enterococci.

Cancer prevention by adult-onset calorie restriction after infant exposure to ionizing radiation in B6C3F1 male mice.

Children are especially sensitive to ionizing radiation and chemical carcinogens, and limiting their cancer risk is of great public concern. Calorie restriction (CR) is a potent intervention for suppressing cancer. However, CR is generally not appropriate for children. This study, therefore, examined to see if adult-onset CR influences the lifetime cancer risk in mice after early-life exposure to ionizing radiation. Infant male mice (1-week-old) were exposed to 3.8 Gy X-rays, fed a control 95 kcal/week or CR 65 kcal/week diet from 7 weeks of age (adult stage), and their lifespan and tumor development were assessed. Irrespective of CR, X-rays shortened lifespan by 38%, and irrespective of irradiation CR extended lifespan by 20%. Thymic lymphoma (TL) and early-occurring non-TL were induced by radiation. The liver and Harderian gland were more susceptible to radiation-induced tumors than the lungs and non-thymic lymphoid tissues (late occurring). CR reduced the risk of hepatocellular carcinoma, late-occurring non-TL, lung tumor, Harderian tumor, and hemangioma but had less impact on TL and early-occurring non-TL. Most notably, the effects of X-rays on induction of lung tumors, late-occurring non-TL and hemangioma were essentially canceled by CR. The ability of CR to prevent late-occurring tumors was the same for non-irradiated and irradiated mice, indicating that the mechanism by which CR influences cancer is independent of irradiation. Our results indicate that adult-onset CR significantly inhibits late-occurring tumors in a tissue-dependent manner regardless of infant radiation exposure.

Mutant prevention concentration of orbifloxacin: comparison between Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus pseudintermedius of canine origin.

BACKGROUND: The mutant prevention concentration (MPC) is an important parameter to evaluate the likelihood of growth of fluoroquinolone-resistant mutants for antimicrobial-pathogen combinations. The MPCs of fluoroquinolones for different canine pathogens have not been compared. In this study, we compared for the first time orbifloxacin MPCs between susceptible strains of Escherichia coli, Pseudomonas aeruginosa, and Staphylococcus pseudintermedius of canine origin. METHODS: More than 1010 CFU/ml of 10 strains of each bacterial species were inoculated onto Muller-Hinton agar supplemented with different concentrations of orbifloxacin from 1× to 64× minimum inhibitory concentration (MIC) and the MPCs were recorded. MICs of original strains and of mutants arising after exposure to sub-MPC concentrations (one per original strain) were determined in the presence or absence of efflux pump inhibitors (EPIs). The effects of quinolone resistance-determining region (QRDR) mutations were also examined. RESULTS: MPCs were significantly higher for P. aeruginosa (16-128 µg/ml) than for E. coli (0.5-32 µg/ml). MPCs for S. pseudintermedius varied between the low-susceptible (16-128 µg/ml) and the high-susceptible strains (4-16 µg/ml) and were the most broadly distributed among the three species. Regarding resistance mechanisms, only one QRDR mutation in gyrA was found in all of the 10 mutants of E. coli and in 4 of the 10 mutants of P. aeruginosa, whereas mutations in both grlA and gyrA were found in 3 mutants and one mutation in grlA was found in 2 mutants among the 10 mutants of S. pseudintermedius. In the presence of an EPI, the MICs of P. aeruginosa mutants decreased markedly, those of E. coli mutants decreased moderately, and those of S. pseudintermedius mutants were unaffected. CONCLUSIONS: MPCs of orbifloxacin vary between bacterial species of canine pathogens, possibly due to the diversity of the main fluoroquinolone resistance mechanism among these species. Therefore, the type of bacterial species should be taken into consideration when using fluoroquinolone drugs such as orbifloxacin in canines.