Molecular and phenotypic characteristics of CMY-2 β-lactamase-producing Salmonella enterica serovar Typhimurium isolated from cattle in Japan

Isolates of extended-spectrum cephalosporin (ESC)-resistant Salmonella enterica serovar Typhimurium obtained from two different farms in Fukushima Prefecture, Japan, in 2007 were characterized in order to determine the genetic basis of resistance. ESC resistance in the two isolates was mediated by an AmpC β-lactamase encoded by the bla(CMY-2) gene, which is located in a large self-transmissible plasmid in each isolate. The sizes of the bla(CMY-2)-carrying plasmids were different. The replicon types of the plasmids were I1-Iγ and A/C. The results of macrorestriction analysis and phage typing suggest a close relationship between both isolates. This is the first report of ESC-resistant S. Typhimurium isolated from cattle in Japan.     

Phylogenetic groups and cephalosporin resistance genes of Escherichia coli from diseased food-producing animals in Japan

A total of 318 Escherichia coli isolates obtained from different food-producing animals affected with colibacillosis between 2001 and 2006 were subjected to phylogenetic analysis: 72 bovine isolates, 89 poultry isolates and 157 porcine isolates. Overall, the phylogenetic group A was predominant in isolates from cattle (36/72, 50%) and pigs (101/157, 64.3%) whereas groups A (44/89, 49.4%) and D (40/89, 44.9%) were predominant in isolates from poultry. In addition, group B2 was not found among diseased food-producing animals except for a poultry isolate. Thus, the phylogenetic group distribution of E. coli from diseased animals was different by animal species. Among the 318 isolates, cefazolin resistance (minimum inhibitory concentrations: ≥32 μg/ml) was found in six bovine isolates, 29 poultry isolates and three porcine isolates. Of them, 11 isolates (nine from poultry and two from cattle) produced extended spectrum β-lactamase (ESBL). The two bovine isolates produced bla_CTX-M-2, while the nine poultry isolates produced bla_CTX-M-25 (4), bla_SHV-2 (3), bla_CTX-M-15 (1) and bla_CTX-M-2 (1). Thus, our results showed that several types of ESBL were identified and three types of β-lactamase (SHV-2, CTX-M-25 and CTX-M-15) were observed for the first time in E. coli from diseased animals in Japan.     

Phylogenetic identification of Cystoisospora spp. from dogs, cats, and raccoon dogs in Japan

Cystoisospora spp. from feces in dogs, cats, and raccoon dogs were isolated, sequenced at the small subunit ribosomal RNA gene locus and compared to other Cystoisospora spp. Cystoisospora oocysts from dogs and raccoon dogs were morphologically similar with those of C. ohioensis, and cat isolates were similar with those of C. felis. The sequences from dogs and raccoon dogs, and cats have a homology with C. ohioensis and C. felis, respectively. Phylogenetic analysis of the DNA sequences showed that the dog and raccoon dog isolates were nested in a clade with other Cystoisospora spp. including C. ohioensis, C. belli, and C. orlovi. The cat isolate formed a sister group with C. felis that was a separate clade from the dog and raccoon dog group. We report sequence variation in these Cystoisospora sequences and have identified raccoon dogs as another carnivore host for Cystoisospora spp. infecting dogs.     

Characterization of monoclonal antibodies against canine P-selectin glycoprotein ligand-1 (PSGL-1)

Thirteen different monoclonal antibodies against canine P-selectin glycoprotein ligand-1 (cPSGL-1) were obtained by immunization of rats with cells of a canine lymphoma cell line (Ema). O-sialoglycoprotein endopeptidase treatment of Ema cells showed that all of these antibodies recognized O-glycosylated peptides of canine PSGL-1. Experiments using deletion or point mutants of cPSGL-1 indicated that these antibodies could be categorized into several groups based on their cPSGL-1 recognition characteristics. These anti-cPSGL-1 monoclonal antibodies will be useful for analysis of the canine P-selectin and PSGL-1 system.     

Molecular complex of lignosulfonic acid/poly(vinyl pyridine) via ionic interaction: characterization of chemical composition and application to material surface modifications

Poly ion complexes of anionic lignosulfonic acid (LSA) with cationic poly(2-vinyl pyridine) or poly(4-vinyl pyridine) were prepared by mixing the polymer solutions in dimethyl sulfoxide or dilute hydrochloric acid. The complexes were formed as soon as the polymer solutions were mixed. Fourier transform infrared (FTIR) measurements for the complexes revealed the presence of strong ionic interactions between sulfonate anions of LSA and pyridinium cations of the vinyl pyridine (VPy) polymers. The strong interaction was supported by detection of a glass transition temperature (T _g) much higher than pristine T _g values of VPy polymers in differential scanning calorimetry. By using the ionic complexation, a coating of LSA onto VPy polymer films was undertaken to modify their original surface property. Formation of a thin complex film was confirmed by observation of absorption bands that are characteristic of the pyridinium cation in reflection FTIR spectra measurements, and by visualization of the attached LSA component in atomic force microscopy. This treatment resulted in the surface of the VPy polymer films becoming hydrophobic. An adhesion test was also conducted by pasting two veneered woods with the polymer complex, followed by shear-tearing them off. The adhesive strength of the complex was estimated to be 1.1 MPa. Part of this report was presented at the 55th (Kyoto, March 2005) and 56th (Akita, August 2006) Annual Meetings of the Japan Wood Research Society     

Ammonia volatilization from Vietnamese acid sulfate paddy soil following application of digested slurry from biogas digester

Paddy and Water Environment - In some areas of Vietnam, digested slurry from biogas digesters is discharged into canals without wastewater treatment. In order to reduce environmental pollution,...     

Oncolytic reovirus synergizes with chemotherapeutic agents to promote cell death in canine mammary gland tumor

The oncolytic effects of reovirus in various cancers have been proven in many clinical trials in human medicine. Oncolytic virotherapy using reovirus for canine cancers is being developed in our laboratory. The objective of this study was to examine the synergistic anti-cancer effects of a combination of reovirus and low doses of various chemotherapeutic agents on mammary gland tumors (MGTs) in dogs. The first part of this study demonstrated the efficacy of reovirus in canine MGTs in vitro and in vivo. Reovirus alone exerted significant cell death by means of caspase-dependent apoptosis in canine MGT cell lines. A single injection of reovirus impeded growth of canine MGT tumors in xenografted mice, but was insufficient to induce complete tumor regression. The second part of this study highlighted the anti-tumor effects of reovirus in combination with low doses of paclitaxel, carboplatin, gemcitabine, or toceranib. Enhanced synergistic activity was observed in the MGT cell line treated concomitantly with reovirus and in all the chemotherapeutic agents except toceranib. In addition, combining reovirus with paclitaxel or gemcitabine at half dosage of half maximal inhibitory concentration (IC₅₀) enhanced cytotoxicity by activating caspase 3. Our data suggest that the combination of reovirus and low dose chemotherapeutic agents provides an attractive option in canine cancer therapy.