Determination of nucleotide sequence of SRY gene in sika deer (Cervus nippon)

The aim of the present study was to determine the whole nucleotide sequence of the open reading frame of the sex‐determining region Y (SRY‐ORF) in wild sika deer. The SRY gene of wild sika deer was obtained by polymerase chain reaction (PCR) with DNA from blood samples. The whole nucleotide sequence of the SRY‐ORF in wild sika deer consisted of 687 bp and encoded 229 deduced amino acids. In comparison with the bovine SRY gene, the percentage of nucleotide sequence homology was 91.0% in the overall ORF, and those of the N‐terminal, high mobility group (HMG) box, and C‐terminal regions within ORF were 88.9%, 96.2% and 87.9%, respectively. The nucleotide sequences of sika deer SRY‐ORF characterized in the present study can be used for phylogenetic analysis or sexing in wild sika deer.     

Daily incremental lines in sika deer (Cervus nippon) dentine

This work was designed to observe the dentine incremental lines of the sika deer (Cervus nippon) fawns and to investigate their periodicity using the chronological labeling method with fluorochromes. The incremental lines were observed in decalcified specimens stained by Bodian's silver technique, and the fluorescence-labeled lines were observed in undecalcified and ground specimens. In the silver stained specimens, there were two types of lines, deeply stained thick lines and faintly stained minute regular incremental lines. The intervals and staining intensities of the deeply stained thick lines were very similar to those of the fluorescence-labeled lines in the ground specimens obtained from the same tooth, and hence, it appeared that the both lines were identical. The number of minute incremental lines between the deeply stained thick lines was the same as that of days between the time when each fluorescent labeling injection was made. Therefore, it seemed that each minute incremental line was formed each day. The possibility of age estimation in days using diurnal dentine increments was discussed.     

Identification and morphological characteristics of dental neonatal line in sika deer (Cervus nippon)

The dental neonatal line of the sika deer (Cervus nippon) was identified experimentally using chronological labeling methods. In the enamel, prominent dark lines were observed under transmitted light, and the number of increments between the dark line and labeling line was almost consistent with the day-age at the time of labeling injection. Therefore, we identified the dark line as the enamel neonatal line. In the dentin, the bright line was observed under polarized light. Since the bright line corresponded to the enamel neonatal line, we recognized the bright line as the dentin neonatal line. Neonatal lines intersected with the enamel-dentin junction at approximately one-third cervical in the first molar. Using these features, it would make possible to distinguish the neonatal line in wild sika deer.     

Hematology in sika deer (Cervus nippon yesoensis Heude, 1884)

Blood samples were taken from 78 wild and 21 farmed sika deer (Cervus nippon yesoensis) using ketamine-xylazine sedation during their excited (82 deer) and resting (17 deer) states. Red cell count (RBC), hemoglobin (Hb), packed cell volume (PCV) and mean corpuscular volume (MCV) were significantly higher and mean corpuscular hemoglobin concentration (MCHC) was lower in excited deer than in resting deer. There was no significant difference in total leukocyte count (WBC) between excited and resting wild males, while a marked increase of WBC with neutrophilia was observed in excited wild females. RBC and PCV were significantly higher and MCH was lower in excited males than in excited females. In wild deer, WBC was significantly higher in females than in males, but there was no significant difference in WBC between farmed males and females. Sex differences in the hematological parameters were not observed in fawns (10 months).     

An occurrence of Salmonella typhimurium infection in sika deer (Cervus nippon)

Seven sika deer (Cervus nippon) in a herd of 30 deer in a park died. Upon examination of three dead deer, Salmonella Typhimurium was isolated from the organs and intestinal contents. Histopathological examination revealed catarrhal enteritis and focal necroses in the liver. Immunohistochemically, Salmonella antigen of O4 was detected in the enteric lesions. The case was diagnosed as S. Typhimurium infection in the sika deer. Because of the importance of Salmonella in public health, fecal and soil samples were continuously collected from the paddock. However, no Salmonella was isolated from any samples collected after medication of the deer and thorough disinfection of the immediate environment.     

Sex determination based on fecal DNA analysis of the amelogenin gene in sika deer (Cervus nippon)

A sex determination method using DNA extracted from feces has been developed for sika deer (Cervus nippon). We determined a partial sequence of the amelogenin gene of sika deer, which exists on both X and Y chromosomes with a deletion region on the Y chromosome. Based on the sexually dimorphic sequences, we designed a pair of primers which could amplify DNA fragments the lengths of which are different between males and females. PCR products were detected in 34 out of 37 fecal samples collected from captured deer and the sexes estimated by the present method were perfectly matched with the actual sexes.     

Chronic malignant catarrhal fever: a case in a sika deer (Cervus nippon)

A condition resembling chronic malignant catarrhal fever was seen in a 9-month-old pet sika hind (Cervus nippon). From an initial acute depression, pyrexia, and anorexia, the condition progressed to include hypopyon, keratitis, lethargy, loss of condition, sloughing of one hoof and eventually death after seven weeks. There were multiple, 5 to 8mm diameter dark-red nodules throughout the mesenteries and mediastinum, along abdominal organ ligaments, and about the uterus and kidneys. Histopathology showed the nodules to be organising vascular thrombi. Concurrent perivascular mononuclear infiltrations and intravascular thrombi in many tissues confirmed that the condition was malignant catarrhal fever.     

Isolation of a Megatrypanum trypanosome from sika deer (Cervus nippon yesoensis) in Japan

A trypanosome was isolated from a sika deer (Cervus nippon yesoensis) in Hokkaido, Japan, during the primary culture of sika deer renal cells. This is the first report of isolation of a Megatrypanum trypanosome from Japanese Cervidae. The trypanosome, designated TSD1, was propagated and maintained in Eagle's modified essential medium containing 20% fetal bovine serum with sika deer renal cells as feeder. The TSD1 trypanosome was morphometrically similar to Trypanosoma cervi, which is commonly isolated from American and European deer. PCR analysis with primers for 18S ribosomal DNA and nucleotide sequencing showed that TSD1 is a member of genus Trypanosoma, subgenus Megatrypanum. Phylogenetically TSD1 is closely related to T. theileri, a common trypanosome of cattle, but is distinguishable from T. theileri by some morphometrical and biological features.     

Intestinal spirochetosis in wild sika deer (Cervus nippon yesoensis) infected with Brachyspira species

Seven adult free-ranging sika deer (Cervus nippon yesoensis) were examined by histology, immunohistochemistry and electron microscopy for intestinal spirochetal infection. Histologically epithelial and goblet cell hyperplasia and edema of the lamina propria mucosa with macrophage and lymphocyte infiltration were observed in the cecum and colon in 6 of the 7 deer. Numerous argyrophilic spirochetes were present in the crypts and some had invaded epithelial and goblet cells and caused degeneration. Immunohistochemically the organisms stained positively with polyclonal antisera against Brachyspira (Serpulina) hyodysenteriae and B. pilosicoli. Ultrastructurally they were 6-14 microm long, 0.2-0.3 microm wide and had 4-6 coils and 13 axial filaments per cell; such features were closely similar to those in the Brachyspira species. These results showed that the spirochetes were capable of inducing enteritis in deer and this intestinal spirochete infection might already be prevalent among wild sika deer in Japan. There is a possibility that this spirochetal colitis is a new syndrome in sika deer and that the same and/or similar spirochetes have infected ruminants, including sika deer and cattle.     

In vivo erythrocyte sickling in the Japanese sika deer (Cervus nippon): methodology.

The Japanese sika deep (Cervus nippon) proved to be a suitable animal model for the study of acute phases of in vivo erythrocyte sickling. Ophthalmologic studies can be conducted during or after 1 to 6 hours of effective in vivo sickling. Intravenous administration of 1.75 to 3.5% sodium bicarbonate solution at a rate of 500 to 1,000 ml per hour produced a transient state of alkalosis and in vivo erythrocyte sickling in the sika deer. The percentage of sickled erythrocytes increased as the blood pH increased. Concurrently, the packed cell volume decreased. Sickling was enhanced by 100% oxygen ventilation after endotracheal intubation and light anesthetization. After the induction of erythrocyte sickling, a sickling-reversal phenomenon occurred despite continued bicarbonate administration. During the course of this reversal, the percentage of sickled erythrocytes steadily decreased, the venous blood pH decreased, and the packed cell volume slowly increased. Because of the sickling-reversal phenomenon, chronic erythrocyte sickling was not achieved.     

Population pharmacokinetics of methadone hydrochloride after a single intramuscular administration in adult Japanese sika deer (Cervus nippon nippon)

ObjectiveTo assess the population pharmacokinetics of methadone in deer.Study designProspective non-randomized experimental trial.AnimalsTwelve healthy adult sika deer (nine males and three females).MethodsDeer received intramuscular administration of racemic methadone hydrochloride at 0.5 mg kg⁻¹ or 1 mg kg⁻¹. Plasma methadone and its metabolite 2-Ethylidene-1,5-Dimethyl-3,3-Diphenyl-Pyrolidine (EDDP) concentrations were determined by validated liquid chromatography coupled to tandem mass spectrometry methods, at times 0, 30 minutes, 1, 2, 3, 4, 5, 6, 8, 12 and 24 hours. Population pharmacokinetics analysis was undertaken using a non-linear mixed effects modelling (NONMEM).ResultsA two-compartment linear disposition model best described observed time-concentration profiles of methadone and EDDP. Population parameter estimates of methadone were elimination clearance (17.3 L hour⁻¹), metabolic clearance (34.6 L hour⁻¹), volume of distribution of compartment 1 (216.0 L) and volume of distribution of compartment 2 (384.0 L). Population parameter estimates of EDDP were elimination clearance (121.0 L hour⁻¹), volume of distribution of compartment 3 (1.08 L) and volume of distribution of compartment 4 (499.5 L). The total clearance and total volume of distribution of methadone and EDDP were 51.9 L hour⁻¹, 121.0 L hour ⁻¹, 600.0 L and 500.6 L, respectively. The methadone terminal elimination half-life was 8.19 hours. No adverse effects were observed after methadone administration.Conclusions and Clinical relevanceFollowing intramuscular injection, methadone was characterized by a large total volume of distribution, high systemic clearance and intermediate terminal half-life in sika deer.     

Detection of Ehrlichia muris DNA from sika deer (Cervus nippon yesoensis) in Hokkaido, Japan

Ehrlichia muris DNA was detected in the blood of sika deer (Cervus nippon yesoensis) by species-specific PCR based on the citrate synthase gene, which was shown to be more sensitive than species-specific PCR based on the 16S rRNA gene. Among 102 deer examined, one deer was positive. Deer may be a possible mammalian reservoir of E. muris.     

Tick infestation of sika deer (Cervus nippon) in the western part of Yamaguchi Prefecture,Japan

Ticks were collected from 94 sika deer (Cervus nippon) hunted in the western part of Yamaguchi Prefecture, Japan from August to November 1999, and March to July 2000. Haemaphysalis longicornis and H. yeni were the dominant species from April to August, while H. flava and H. megaspinosa were dominant in October, November and March. This is the first report of H. yeni in the mainland of Japan. Small numbers of H. kitaokai, Amblyomma testudinarium and Ixodes ovatus were also recorded.     

Ophthalmic manifestations of induced sickling of erythrocytes in Japanese sika deer (Cervus nippon).

Nine female Japanese sika deer (Cervus nippon) were used in a total of 25 experiments in which sickling was chemically induced. During these experiments, color fundic and color fluorescein photographs were taken. Fundic changes included retinal vascular attenuation, blood column pallor, and decreased tapetal reflectivity. These changes were most likely directly associated with a decreased hematocrit and a generalized shocklike condition. Three deer had a congested appearance in retinal blood vessels and tapetum lucidum. Two of the 3 deer developed serous detachment of the retina. These changes seemingly were associated with severe venous statis; all 3 deer died shortly after the experiment was terminated. These experiments yielded data only for the acutely affected deer. None of the ocular changes could be considered the result of chronic sickling because of the reversal of sickling that occurred despite continued intravenous administration of bicarbonate. None of the deer developed ocular changes characteristic of sickle cell retinopathy in human beings. The changes in human beings probably result from continued stress and prolongation of sickling, and especially from a multiplicity of repeated severe episodes of sickling occurring over many years.     

Seasonal changes of the mineral contents in the rumen of wild Yeso sika deer (Cervus nippon yesoensis)

The rumen contents were collected from 36 wild Yeso sika deer (Cervus nippon yesoensis) captured by deer culling or by hunting in the spring, summer, autumn and winter in Hokkaido, Japan. Botanical classification was conducted, and the contents of mineral (calcium (Ca), phosphorus (P), potassium (K), sodium (Na), iron (Fe), copper (Cu) and zinc (Zn)) were measured. The animals were captured around pastures or fallow field areas in the Kushiro area. The rumen contents consisted of grasses and Sasa sp. leaves regardless of the season. Leaves and bark were ingested in the spring, autumn and winter. The macro‐mineral contents in the rumen showed seasonal changes. In the summer, the Ca, K and P contents were high, and the Na content was low. There were no seasonal changes in the Fe content. The P, Na and Fe contents were higher than the animals' requirements. In a future survey, it is needed to determine the mineral contents of the food ingested by wild Yeso sika deer.     

Effects of adding a commercial capsaicin fertilizer on the feeding behavior of sika deer (Cervus nippon)

We investigated the inhibitory effect of capsaicin fertilizer on feeding in deer. We tested four captive adult female deer. In Experiment 1, in addition to the treatment (intact) containing only a solid feed (HC), we mixed the fertilizer not containing capsaicin (F) or the capsaicin fertilizer (CF) in the solid feed. In addition, the solid feed was put on a wire net that capsaicin fertilizer was placed 5 cm below (SCF). We investigated their feeding behavior response. In Experiment 2, we changed the amount of substance (fertilizer and capsaicin fertilizer) mixed in the HC. We mixed different amounts (0, 50, 100, and 200 g) of the treatments other than the intact with HC and presented them to the deer, and investigated their feeding behavior response. In Experiment 1, intake in the F and CF decreased (p < .05). In Experiment 2, HC intake was significantly lower in the 100 and 200 g CF (p < .05). However, HC intake relatively increased by the last day in the CF 200 g too. The capsaicin fertilizer decreased the feeding behavior of deer by directly touching the mucous membranes of the deer nose and lips. However, the effects were decreased over time.     

Occurrence of two distinct Theileria lineages in sika deer (Cervus nippon) of Iwate Prefecture, Japan

In the present study, we tried to detect protozoan blood parasites from the liver or blood of 141 Japanese sika deer (Cervus nippon) in Iwate Prefecture of Japan by polymerase chain reaction. Approximately 500-bp amplicons were obtained in 76 (53.9%) of 141 samples by amplification for V4 hyper-variable regions of the 18S rRNA gene, and the amplicons were considered to be DNA of Theileria species. The complete nucleotide sequences (1701-bp) of the 18S rRNA gene were determined in 25 samples and were divided into 8 genotypes that were phylogenetically separated into two distinct lineages showing a monophyletic relation. The two lineages of Theileria were detected in different rates (12 and 88%) from sika deer in Iwate Prefecture.