Sex-inducing effects toward planarians widely present among parasitic flatworms.

Various parasitic flatworms infect vertebrates for sexual reproduction, often causing devastating diseases in their hosts. Consequently, flatworms are of great socioeconomic and biomedical importance. Although the cessation of parasitic flatworm sexual reproduction is a major target of anti-parasitic drug design, little is known regarding bioactive compounds controlling flatworm sexual maturation. Using the planarian Dugesia ryukyuensis, we observed that sex-inducing substances found in planarians are also widespread in parasitic flatworms, such as monogeneans and flukes (but not in tapeworms). Reverse-phase HPLC analysis revealed the sex-inducing substance(s) eluting around the tryptophan retention time in the fluke Calicophoron calicophorum, consistent with previous studies on the planarian Bipalium nobile, suggesting that the substance(s) is likely conserved among flatworms. Moreover, six of the 18 ovary-inducing substances identified via transcriptome and metabolome analyses are involved in purine metabolism. Our findings provide a basis for understanding and modifying the life cycles of various parasitic flatworms.

Genotypic and Phenotypic Features of Eye-Origin Streptococcus canis Isolates from Dogs in 2021: Relatedness with Clonal Complex 46 and Antimicrobial Resistance.

The eye (including the cornea) and ear canal are the major sources of Streptococcus canis in companion animal practice. In this study, we aimed to clarify the genotypic and phenotypic features of eye-origin isolates collected in 2021 compared to ear-origin isolates collected in 2021 and eye-origin isolates collected in 2017. Of the 102 isolates in 2021, 9 eye-origin isolates were enrolled. Twenty ear-origin isolates in 2021 and 13 eye-origin isolates in 2017 were included as controls. Genotypic analyses included profiling of virulence-associated genes (VAGs; inl, sagA, slo, scp, lbp, fbp, gbp, ap1, fp1, and brp), S. canis M-like protein (SCM) allele typing, multilocus sequence typing, and antimicrobial resistance (AMR) genotyping and phenotyping analyses including hemolytic activity (HA) measurement and AMR phenotyping. One 2017-eye-origin isolate displayed high-level HA; the others displayed low-level HA. No association was evident between the 2021-eye-origin population and the detection rate of each VAG. There was no association between the 2021-eye-origin population and the main SCM allele 2. A significant association was evident between the 2021-eye-origin population and the main clonal complex (CC) 46 containing sequence type (ST) 46/ST2. A significant association was also detected between the 2021-eye-origin population and AMR phenotypes/genotypes. Our observations suggest unique microbiological features (CC46 with AMR phenotypes/genotypes) among the 2021-eye-origin population.

Draft Genome Sequence of Blood-Origin Pasteurella canis Strain PA42, Isolated from a Dog in Japan.

We report the draft genome sequence of Pasteurella canis strain PA42, which was isolated from the blood of a diseased dog in Japan in 2021. The 2.151-Mbp genome has a G+C content of 36.6%. Sequences unmapped to the reference genome sequence of NCTC 11621T (GenBank accession number UGTV00000000.1) were characterized.

Reduced rates of antimicrobial resistance in Staphylococcus intermedius group and Escherichia coli isolated from diseased companion animals in an animal hospital after restriction of antimicrobial use.

The 2016 National Action Plan aims for reduction in antimicrobial resistance (AMR) to tetracyclines, third-generation cephalosporins, and fluoroquinolones in Escherichia coli isolates from livestock: to lower the tetracycline resistance of E. coli to 33% or less; to maintain the third-generation cephalosporin resistance of E. coli at the same level as in the other G7 countries as of 2020; and to maintain the fluoroquinolone resistance of E. coli at the same level as in the other G7 countries as of 2020. A relatively unexplored facet of reducing AMR is the impact of minimizing transmission of AMR strains by companion animals. In this study we compared AMR rates in Staphylococcus intermedius group (SIG) and E. coli isolated from diseased companion animals in an animal hospital before and after restriction of antimicrobial use. Our study spanned a 4.5-year period from 2014 to June 2018 during which antimicrobial use was restricted in 2016. During this period, abundance of methicillin-resistant SIG isolates from the hospital dropped from 41.5% to 9.3%, and that of extended-spectrum ß-lactamase (ESBL)-producing E. coli isolates dropped from 29.5% to 9.5%. Tests for antimicrobial susceptibility revealed significantly reduced rates of AMR to enrofloxacin and levofloxacin in SIG isolates, and to cefazolin in E. coli isolates after antimicrobial use was restricted. Our observations suggest that restriction of antimicrobial use, especially that of third-generation cephalosporins and fluoroquinolones, is an effective method for reducing AMR rates. These findings will be relevant in guiding antimicrobial restriction approaches in other animal hospitals and clinics.

Bacteria isolated from companion animals in Japan (2014-2016) by blood culture.

We aimed to identify microorganisms isolated by blood culture (BC) from companion animals and to determine antimicrobial resistance of these isolates during 2014-2016 at veterinary laboratory, in comparison with those during 2010-2013, in Japan. Clinical data (animal species, visiting animals/hospitalized animals, and others except for disease type and clinical course including history of antimicrobial agent use) on ill animals at veterinary clinics or hospitals were obtained. We retrospectively analyzed animal-origin BC results extracted from the database in 2014-2016 and those obtained in 2010-2013. BC-positive samples were from most of dogs (n = 174 in 2014-2016 and n = 86 in 2010-2013). Escherichia coli (n = 50, 25.1%) and Staphylococcus intermedius group (SIG) bacteria (n = 23, 11.6%) were most prevalent in 2014-2016, while the percentages of E. coli (n = 22, 25.3%) and SIG (n = 9, 10.3%) in 2010-2013 were similar to those in 2014-2016. Percentages of extended-spectrum ß-lactamase (ESBL)-producing E. coli and methicillin-resistant staphylococci (MRS) rate of SIG bacteria isolated in 2014-2016 were 28.0% and 69.6% (vs. 22.7% and 44.4% in 2010-2013), respectively. Fourteen ESBL-producing E. coli in 2014-2016 were isolated from 7 visiting animals and 7 hospitalized ones, whereas the sixteen MRS of SIG were from 7 visiting animals and 9 hospitalized ones. Our observations support the prevalent microorganisms isolated by BC and their antimicrobial resistance patterns for two study periods.

Identification of Group G Streptococcal Isolates from Companion Animals in Japan and Their Antimicrobial Resistance Patterns.

In this study, we conducted a species-level identification of group G streptococcal (GGS) isolates from companion animals in Japan and analyzed antimicrobial resistance (AMR) patterns. Strains were isolated from sterile and non-sterile specimens collected from 72 animals with clinical signs or symptoms in April-May, 2015. We identified the strain by 16S rRNA sequencing, mass spectrometry (MS), and an automated method based on their biochemical properties. Antimicrobial susceptibility was determined using the broth microdilution method and E-test. AMR determinants (erm(A), erm(B), mef(A), tet(M), tet(O), tet(K), tet(L), and tet(S)) in corresponding resistant isolates were amplified by PCR. The 16S rRNA sequencing identified the GGS species as Streptococcus canis (n = 68), Streptococcus dysgalactiae subsp. equisimilis (n = 3), and S. dysgalactiae subsp. dysgalactiae (n = 1). However, there were discrepancies between the sequencing data and both the MS and automated identification data. MS and the automated biochemical technique identified 18 and 37 of the 68 sequencing-identified S. canis strains, respectively. The AMR rates were 20.8% for tetracycline and 5.6% for clarithromycin, with minimum inhibitory concentrations (MIC)50 -MIC90 of 2-64 and ≤ 0.12-0.25µg/mL, respectively. AMR genotyping showed single or combined genotypes: erm(B) or tet(M)-tet(O)-tet(S). Our findings show the unique characteristics of GGS isolates from companion animals in Japan in terms of species-level identification and AMR patterns.

A study on Borna disease virus infection in domestic cats in Japan.

Borna disease virus (BDV) infection causes neurological disease in cats. Here, we report BDV infection in 199 hospitalized domestic cats in the Tokyo area. BDV infection was evaluated by detection of plasma antibodies against BDV-p24 or -p40. BDV-specific antibodies were detected in 54 cats (27.1%). Interestingly, the percentage of seropositive cats was not significantly different among the three clinical groups, i.e., healthy (29.8%), neurologically asymptomatic disease (22.2%) and neurological disease (33.3%). The specific antibodies were present even in cats aged below one year. The seropositive ratio was constant, irrespective of age and sampling season. The present study suggests that additional factors are required for onset of Borna disease in naturally infected cats and that BDV is transmitted through vertical routes in cats.

Large-scale survey of adverse reactions to canine non-rabies combined vaccines in Japan.

Canine non-rabies combined vaccines are widely used to protect animals from infectious agents, and also play an important role in public health. We performed a large-scale survey to investigate vaccine-associated adverse events (VAAEs), including anaphylaxis, in Japan by distributing questionnaires on VAAEs to veterinary hospitals from April 1, 2006 through May 31, 2007. Valid responses were obtained for 57,300 vaccinated dogs at 573 animal hospitals; we obtained VAAEs information for last 100 vaccinated dogs in each veterinary hospital. We found that of the 57,300, 359 dogs showed VAAEs. Of the 359 dogs, death was observed in 1, anaphylaxis in 41, dermatological signs in 244, gastrointestinal signs in 160, and other signs in 106. Onset of VAAEs was mostly observed within 12h after vaccination (n=299, 83.3%). In this study, anaphylaxis events occurred within 60 min after vaccination, and about half of these events occurred within 5 min (n=19, 46.3%). Furthermore, where anaphylaxis was reported, additional information to support the diagnosis was obtained by reinvestigation. Our resurvey of dogs with anaphylaxis yielded responses on 31 dogs; 27 of these demonstrated collapse (87.1%), 24 demonstrated cyanosis (77.4%), and both signs occurred in 22 (71.0%). Higher rates of animal VAAEs, anaphylaxis, and death were found in Japan than in other countries. Further investigations, including survey studies, will be necessary to elucidate the interaction between death and vaccination and the risk factors for VAAEs, and thus develop safer vaccines. Moreover, it may also be necessary to continually update the data of VAAEs.