Isolation of exfoliative toxin from Staphylococcus intermedius and its local toxicity in dogs

A rounding effect was demonstrated in cultured cells inoculated with the culture filtrates (CFs) of 60 strains of Staphylococcus intermedius derived from dogs affected with pyoderma. Exfoliative toxin (ET)-like toxin (ETLT) was isolated from the CF of S. intermedius strain D-52, which exhibited strong rounding activity and then was purified by gel filtration on a Sephadex G-75 column, and by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The ETLT caused exfoliation in 1-day-old chickens, suckling Syrian hamsters, and dogs, but not in suckling mice. The ETLT was serologically different from exfoliative toxin A (ETA), exfoliative toxin B (ETB), exfoliative toxin C (ETC), S. hyicus exfoliative toxin A (SHETA), and SHETB, as shown by Western blot analysis. The molecular weight of the ETLT was estimated at 30 kDa by SDS-PAGE. In the present study, we propose the ETLT was a novel type of ET, S. intermedius exfoliative toxin (SIET).     

Fenprostalene-induced abortion in bitches

A long-acting prostaglandin F(2alpha) analogue, fenprostalene, was subcutaneously administered singly at doses of 5, 10, 25, 50, 100 or 150 microg on day 25 after ovulation during pregnancy in bitches (n=4 for each dosage), and maintenance of pregnancy, changes in plasma progesterone concentration, interval between the treatment and subsequent estrus and conception rate at that estrus were studied. Abortion was associated with a decrease in the plasma progesterone concentration below about 2 ng/ml, and the abortifacient effect was dose-dependent. Administration of 50 microg or more of fenprostalene induced abortion in all dogs 3 to 13 days after the treatment. The interval between administration and subsequent estrus was markedly shorter in the aborted bitches than in the non-aborted ones (P<0.01). The conception rate at the estrus in the aborted dogs was 50%, whereas all of the bitches who had not aborted became pregnant. The results indicate that a single administration of fenprostalene could induce abortion during mid-pregnancy in bitches, and the subsequent estrus may come early with a low conception rate.     

Ammonia-assimilating microbes in microbial community in a lagoon for wastewater from paddock of dairy cattle

In the present article the distribution and abundance of ammonia‐assimilating microbes among the natural habitants in a lagoon were investigated. In the medium containing lagoon‐extract, about 20% of total 82 isolates showed ammonia‐assimilating ability. By sequencing of 16S ribosomal DNA (rDNA), the highest ammonia‐assimilating isolates at 10°C and 37°C were identified as Janthinobacterium lividum and Bacillus sp., respectively. The structure of the microbial community was analyzed by polymerase chain reaction‐denaturing gradient gel electrophoresis (PCR‐DGGE). Almost of the dominant species that were detected by PCR‐DGGE did not coincide with isolates, which showed the high ammonia‐assimilating ability, but one specie by PCR‐DGGE coincided with ammonia‐assimilating isolate. These results suggested that ammonia‐assimilating microbes existed as non‐dominant species in the microbial community in a lagoon.     

Estrous cycle based on blood progesterone profiles and changes in vulvar appearance of Malayan tapirs (Tapirus indicus)

The progesterone (P(4)) profiles and macroscopic vulvar changes of female Malayan tapirs were investigated in order to understand their fundamental reproductive physiology and to search for visual indicators of estrus. Blood was collected once or twice a week from seven female Malayan tapirs kept at four zoos. Serum or plasma P(4) concentrations were determined by radioimmunoassay. The P(4) concentrations changed cyclically throughout the years, and a total of 56 cycles was confirmed in the seven females. The length of the estrous cycle based on the P(4) profiles was 43.6+/-2.0 days; however, this mean includes great variation in length, from 21 to 84 days. Mucous discharge from the vulva and vulvar swelling were seen when the P(4) concentrations were low before the beginning of a rise in most cases. In conclusion, captive female Malayan tapirs have variations of approximately 1 to 3 months in estrous cycle length, and visual changes in the vulva are helpful in estimating estrus in female Malayan tapirs.     

Glutathione supplementation to semen extender improves the quality of frozen-thawed canine spermatozoa for transcervical insemination

The present study was conducted to evaluate whether supplementation of semen extender with glutathione (GSH) can maintain the quality of frozen-thawed canine spermatozoa. Eighteen ejaculates were obtained from 5 dogs and placed in extender (20% egg yolk, Tris, citric acid, lactose, raffinose, antibiotics and 6.5% glycerol) containing 0 (control), 2.5, 5, 7.5 or 10 mM GSH. The samples were cooled to 4 C and then frozen in liquid nitrogen vapor. Motility parameters of the sperm were evaluated at 0, 1, 2, 3, 4, 12 and 24 h after thawing. Sperm motility was higher in the 5 mM GSH group than in the control or 2.5 and 10 mM GSH groups; this effect was observed at 1 to 24 h after thawing (P < 0.05). The 5 mM GSH group had a higher sperm viability index at 12 and 24 h after thawing compared with the other groups (P < 0.05). Acrosome integrity, evaluated at 4 h after thawing, was greater in two of the GSH-treated groups (5 and 10 mM) compared with the control. Lipid peroxidation (LP) levels immediately after thawing were lower in the 5 and 10 mM GSH groups compared with the control, while those at 12 h after thawing did not differ significantly. Frozen-thawed semen in the 5 mM GSH group was used for transcervical insemination of 4 bitches, resulting in delivery of 5 puppies from 2 bitches. These results indicate that supplementation of semen extender with 5 mM GSH was effective in improving motility, longevity and acrosomal integrity and inhibiting LP levels in post-thaw canine spermatozoa, without any adverse impacts on full-term development after transcervical insemination.     

Stromal cell-derived factor-1 (SDF1)-dependent recruitment of bone marrow-derived renal endothelium-like cells in a mouse model of acute kidney injury

Ischemic acute kidney injury (AKI) is the most key pathological event for accelerating progression to chronic kidney disease through vascular endothelial injury or dysfunction. Thus, it is critical to elucidate the molecular mechanism of endothelial protection and regeneration. Emerging evidence indicates that bone marrow-derived cells (BMCs) contribute to tissue reconstitution in several types of organs post-injury, but little is known whether and how BMCs contribute to renal endothelial reconstitution, especially in an early-stage of AKI. Using a mouse model of ischemic AKI, we provide evidence that incorporation of BMCs in vascular components (such as endothelial and smooth muscle cells) becomes evident within four days after renal ischemia and reperfusion, associated with an increase in stromal cell-derived factor-1 (SDF1) in endothelium and that in CXCR4/SDF1-receptor in BMCs. Notably, anti-CXCR4 antibody decreased the numbers of infiltrated BMCs and BMC-derived endothelium-like cells, but not of BMC-derived smooth muscle cell-like cells. These results suggest that reconstitution of renal endothelium post-ischemia partially depends on a paracrine loop of SDF1-CXCR4 between resident endothelium and BMCs. Such a chemokine ligand-receptor system may be attributable for selecting a cellular lineage (s), required for renal vascular protection, repair and homeostasis, even in an earlier phase of AKI.     

Acquired Fanconi syndrome in a dog exposed to jerky treats in Japan

A 6-year-old spayed female Jack Russell Terrier presented with a 1-month history of lethargy, anorexia, vomiting and weight loss. The dog was fed beef and chicken jerky treats daily in addition to a commercial diet. Laboratory tests revealed azotemia, hypokalemia, hyperchloremia, metabolic acidosis and glucosuria with normoglycemia. Urine amino acid analysis showed significant amino acid loss into the urine. Thus, Fanconi syndrome was diagnosed, and based on the case history and extensive diagnostic testing, excessive consumption of jerky treats was strongly suspected as the cause. Glucosuria resolved 7 days after the withdrawal of jerky treats and fluid therapy. Aminoaciduria was substantially, but not completely, improved 3 months after diagnosis. Mild azotemia remained, suggesting chronic renal disease. To the best of our knowledge, this is the first reported case of Fanconi syndrome following the consumption of jerky treats in Japan.