Antifungal activities of the combination of tacrolimus and itraconazole against Trichophyton mentagrophytes

Tacrolimus was reported to have no antifungal activity against dermatophytes, but it might show a synergistic fungicidal activity with azoles against dermatophytes as in the case of pathogenic yeasts. Therefore, we examined the antifungal activity of tacrolimus combined with itraconazole (ITZ) against five strains of Trichophyton mentagrophytes by measuring cell growth and performing MIC (minimum inhibitory concentration) testing. The mean colony diameter of T. mentagrophytes on 1/10 diluted Sabouraud dextrose agar (dSDA) with tacrolimus combined with ITZ was significantly shorter than that on dSDA with tacrolimus or ITZ. MICs of tacrolimus, ITZ, and tacrolimus combined with ITZ against T. mentagrophytes were determined, respectively. The results revealed a synergistic activity of tacrolimus in combination with ITZ against T. mentagrophytes.     

Detection of rickettsial DNA in ixodid ticks recovered from dogs and cats in Japan

DNA from ticks recovered from 1137 dogs and 133 cats from all over Japan were examined for Rickettsia infection by citrate synthase gene (gltA)-based PCR and partial nucleotide sequencing. A total of 91 dog tick samples and 18 cat tick samples showed a single band of the appropriate size in the nested PCR. Sequence analysis was successfully performed on 102 samples. DNA of Rickettsia japonica or closely related Rickettsia spp. strains were detected from 38 ticks in 16 prefectures mainly in western Japan. The other 33, detected from 13 prefectures including Hokkaido and Okinawa, were found to be Rickettsia helvetica or closely related strains. A total of 29 DNA that showed highest homology with Rickettsia akari or closely related strains were detected in 19 prefectures, widespread throughout Japan. Rickettsia canada-like DNA was detected from Haemaphysalis sp. removed from a dog in Fukuoka, and ;Candidatus Rickettsia tarasevichiae'-like DNA was from Ixodes sp. removed from a dog in Hokkaido.     

Effects of initial handling and training on autonomic nervous function in young Thoroughbreds

OBJECTIVE: To determine the effects of initial handling and training on autonomic nervous functions in young Thoroughbreds. ANIMALS: 63 healthy Thoroughbreds. PROCEDURE: All horses were trained to be handled and initially ridden in September of the yearling year and then trained until the following April by conventional training regimens. To obtain the heart rate (HR), electrocardiograms were recorded in the stable before initial handling and training and following 7 months of training; variations in HR were then evaluated from the power spectrum in terms of the low frequency (LF; 0.01 to 0.07 Hz) power and high frequency (HF; 0.07 to 0.6 Hz) power as indices of autonomic nervous activity. To evaluate the fitness, the V200 (velocity at HR of 200 beat/min), which is reflective of the aerobic capacity of the horse, was measured. RESULTS: Mean (+/- SE) resting HR decreased significantly from 41.5 +/- 0.8 to 38.7 +/- 0.4 beat/min following 7 months of training. The LF power of horses increased significantly from 1,037 +/- 128 milliseconds2 in September of the yearling year to 2,944 +/- 223 milliseconds2 in the following April. Similarly, the HF power increased significantly from 326 +/- 30 milliseconds2 to 576 +/- 39 milliseconds2 at the corresponding time points. The V200 increased significantly following training. CONCLUSIONS AND CLINICAL RELEVANCE: Increases in LF and HF powers indicate that parasympathetic nervous activity increases in horses by 7 months of training. The decrease in resting HR may be dependent on the training-induced increase of parasympathetic nervous activity in Thoroughbreds.     

Light-chain multiple myeloma in a cat.

A diagnosis of light-chain multiple myeloma was made in an 11-year-old male American Shorthair cat. The cat showed atypical plasma cell infiltration in the bone marrow, biclonal gammopathy caused by polymerization of myeloma protein (M-protein), and Bence-Jones proteinuria. The M-protein in the serum of the cat was analyzed by using 12% sodium dodeyl sulfate (SDS) polyacrylamide gel electrophoresis with Coomassie brilliant blue staining. An intense band with a size of 27 kDa, the size of the immunoglobulin light chain, was clearly observed, whereas the band corresponding to the immunoglobulin heavy chain (59 kDa) was undetectable. The 27-kDa band was confirmed to be an immunoglobulin light chain by Western blotting by using antibodies for feline immunoglobulin. These data suggested that the neoplastic plasma cells produce light chain only, leading to the diagnosis of light-chain multiple myeloma in the cat.     

Neuroendocrine mechanism mediating energetic regulation of gonadotropin release in female rats

Energy level is a critical factor controlling gonadal activity at various phases of reproduction. A female rat model has revealed that fasting‐induced luteinizing hormone (LH) suppression is mediated by a specific neural pathway, such as noradrenergic neurons originating in the A2 region and projecting to the hypothalamic paraventricular nucleus and corticotropin‐releasing hormone neurons. The pathway is shared with that mediating glucoprivic suppression of LH pulses. Among the peripheral signals altered by energy deficiency, glucose could be a signal molecule conveying the peripheral information to the brain to regulate feeding and gonadotropin‐releasing hormone/LH release through the noradrenergic pathway during undernutrition. The brain detects the energy availability to control feeding and reproductive function at various phases of an animal’s life. It is most likely that the central glucose‐sensing mechanism could be similar to the pancreatic one, involving a glucokinase‐mediated process to detect glucose availability. Further studies are needed to elucidate the mechanism integrating the energy signals.     

Combined administration of ghrelin and GHRH synergistically stimulates GH release in Holstein preweaning calves

Ghrelin is a gut peptide which participates in growth regulation through its somatotropic, lipogenic and orexigenic effects. Synergism of ghrelin and growth hormone-releasing hormone (GHRH) on growth hormone (GH) secretion has been reported in humans and rats, but not in domestic animals in vivo. In this study, effects of a combination of ghrelin and GHRH on plasma GH and other metabolic parameters, and changes in plasma active and total ghrelin levels were studied in Holstein bull calves before and after weaning. Six calves were intravenously injected with vehicle (0.1% BSA-saline), ghrelin (1 microg/kg BW), GHRH (0.25 microg/kg BW) or a combination of ghrelin plus GHRH at the age of 5 weeks and 10 weeks (weaning at 6 weeks of age). Ghrelin stimulated GH release with similar potency as GHRH and their combined administration synergistically stimulated GH release in preweaning calves. After weaning, GH responses to ghrelin and GHRH became greater compared with the values of preweaning calves, but a synergistic effect of ghrelin and GHRH was not observed. The GH areas under the concentration curves for 2h post-injection were greater in weaned than in preweaning calves (P<0.05) if ghrelin or GHRH were injected alone, but were similar if ghrelin and GHRH were injected together. Basal plasma active and total ghrelin levels did not change around weaning, but transiently increased after ghrelin injection. Basal plasma insulin, glucose and non-esterified fatty acid levels were reduced after weaning, but no changes by treatments were observed. In conclusion, ghrelin and GHRH synergistically stimulated GH release in preweaning calves, but this effect was lost after weaning.     

A case of adenocarcinoma of the endometrium extending into the leiomyoma of the uterus in a rabbit

In a pet rabbit, 2 tumor masses one on each horn were macroscopically seen in the wall of the uterus. On light microscopic examination, the right horn mass consisted of an admixture of neoplastic epithelial and mesenchymal element. The epithelial element was composed of neoplastic epithelial cells with numerous mitotic figures and formed varied sizes of acini, glandular, and solid structures. The tumor was diagnosed as an adenocarcinoma of the endometrium. The mesenchymal element was composed of well-differentiated smooth muscle cells and was diagnosed as a leiomyoma. While adenocarcinoma cells formed a protrusive mass in the uterine lumen, they also showed an extension into the leiomyoma of the myometrium. By immunohistochemistry, adenocarcinoma stained positive for cytokeratin (MNF116) and leiomyoma stained positive for smooth muscle actin, showing a substantial difference in the cytological nature of these tumor cells. The results may give a further evidence supporting the narrative of the tumor development that an adenocarcinoma of the endometrium extended into leiomyoma of the uterus. To the author's knowledge, this is the first report describing this type of combination of two independent tumors in a pet rabbit.     

Diluted isoflurane as a suitable alternative for diethyl ether for rat anaesthesia in regular toxicology studies

Despite its explosive properties and toxicity to both animals and humans, diethyl ether is an agent long used in Japan in the anaesthesia jar method of rat anaesthetises. However, in response to a recent report from the Science Council of Japan condemning diethyl ether as acceptable practice, we searched for an alternative rat anaesthesia method that provided data continuous with pre-existing regular toxicology studies already conducted under diethyl ether anaesthesia. For this, we examined two candidates; 30% isoflurane diluted with propylene glycol and pentobarbitone. Whereas isoflurane is considered to be one of the representatives of modern volatile anaesthetics, the method of propylene glycol-diluted 30% isoflurane used in this study was our modification of a recently reported method revealed to have several advantages as an inhalation anaesthesia. Intraperitoneal pentobarbitone has long been accepted as a humane method in laboratory animal anaesthesiology. These 2 modalities were scrutinized in terms of consistency of haematology and blood chemistry with previous results using ether. We found that pentobarbitone required a much longer induction time than diethyl ether, which is suspected to be the cause of fluctuations in several haematological and blood chemical results. Conversely, only calcium ion concentration showed a slight difference from traditional results in the case of 30% isoflurane. Additionally, serum prolactin and corticosterone levels indicated that 30% isoflurane induced less stress than ether, confirming that 30% isoflurane can both provide results consistent with diethyl ether, while at the same time remove its disadvantages. As such 30% isoflurane appears to be a strong alternative anaesthetic agent for future regular toxicology studies in Japan.     

Changes of multi-drug resistance pattern in Salmonella enterica subspecies enterica serovar typhimurium isolates from food-producing animals in Japan

A total of 153 isolates of Salmonella Typhimurium derived from food-producing animals in Japan between 2002 and 2005 were investigated for antimicrobial resistance and phage types related to definitive phage type 104 (DT104). The predominant resistance type was resistance to ampicillin, dihydrostreptomycin, kanamycin, and oxytetracycline in bovine (45.2%, 48/104) and resistance to dihydrostreptomycin and oxytetracycline in porcine isolates (58.7%, 27/48). DT104-related phage type was found in 32 of 104 bovine isolates, two of 48 porcine isolates, and one of eight isolates from poultry, showing that the proportion of the phage type in S. Typhimurium isolates from cattle and pigs significantly (P<0.01) decreased from 71.9% and 31.4% in 1999-2001 to 30.8% and 4.1% in 2002-2005, respectively.     

Genomic organization of the T-cell receptor gamma gene and PCR detection of its clonal rearrangement in canine T-cell lymphoma/leukemia

Because the T-cell receptor gamma (TCRgamma) gene is rearranged at an early stage of T-cell development in both TCRalphabeta and TCRgammadelta lineages, it has been preferentially targeted to detect T-cell clonality in human lymphoma/leukemia. We isolated 22 independent cDNA clones encoding canine TCRgamma and the following analysis of nucleotide sequences using the dog genome database revealed that the canine TCRgamma locus contains at least four repertories of variable genes that can be organized into two distinct subgroups and six repertories of joining genes belonging to two distinct subgroups according to the nucleotide sequence similarity. The findings allowed us to design PCR primers that were directed to the conserved or specific nucleotide sequences for each subgroup of variable and joining genes. By using four different combinations of primers, a PCR-based analysis was performed on cell samples collected from T-cell lymphoma/leukemia and B-cell lymphoma cases and hyperplastic and normal lymph nodes. All cell samples from 11 T-cell malignancy cases exhibited clonal amplification by two out of four primer combinations. This finding was considered to be valuable in PCR-based analysis for detecting T-cell clonality in canine lymphoma/leukemia.