Some young large farm animals show a laminar bone formation in the long-bone cortex. Such a laminar bone is gradually replaced by Haversian bone with osteons during their growth periods. In this preliminary study, we observed the transverse ground samples of tibia cortex in young calves, pigs, and sheep by backscattered electron imaging. The cortex bones of all the newborn (NB) animals were basically formed with laminar bone structures. The NB and 1-month-old (1-M) calves had a typical concentric structure of laminar bone, whereas the NB and 1-M pigs showed a wire-netting bone with laminar-bone units. The NB sheep was similar to the calf rather than the pig. In the growth rate of bone volume, sheep was similar to calf up to 6 months after birth (6-M). Such calf and sheep showed a more rapid ratio of bone volume than pig. A few osteons had initially appeared in the innermost layer of the 6-M calf. A 1-year-old (1-Y) calf showed scattered osteons in the bone cortex, but many laminar-bone units were still retained in the outer layer. A 6-M pig had many osteons in the entire cortex but only a few osteons in the outermost layer. In the 6-M sheep, no osteons were observed, whereas a 1-Y sheep showed a relatively small number of osteons mainly in the middle layer but a higher osteon-volume than the 1-Y calf. In the 1-Y sheep, the more widely absorbed areas by bone-remodeling with osteons were observed as compared with the 1-Y calf, and the bone volume was decreased from the 6-M into the 1-Y sheep because of the remarkable bone-absorption. Thus, calf kept on possessing many laminar-bone units for a longer time in the growth period than sheep, while pig showed the earliest bone-remodeling with osteons. These results may be caused by their different body size and withers height in calf and sheep after growing and the difference of the dependence upon mother's body during juvenile period between pig and calf with sheep. The initial region of osteon formation may be distinguishable among their animals, respectively. However, further detailed investigations of their young animals at successive stages will be necessary.

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A single olivocerebellar fiber branches off several climbing fibers. One Purkinje cell receives input from only one climbing fiber. A single inferior olivary neuron, therefore, synapses with several Purkinje cells, so that there are more Purkinje cells than the inferior olivary neurons. We aimed to elucidate the numerical ratio of the inferior olivary neurons to Purkinje cells in the chicken. The total numbers were 353,834 ± 5,274 in the Purkinje cells per the cerebellum and 21,553 ± 904 in the inferior olivary neurons of both sides. The numerical ratio of inferior olivary neurons to Purkinje cells was 1:16. The ratio of those neurons in mammals is about 1:4-17, so that the ratio in the chicken is within the range of mammals.

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One hundred thirty-seven broiler chickens at a poultry meat processing plant had dark green to black livers. Thirty-one chickens of these were collected at random and examined pathologically and biochemically. All of thirty-one chickens were female. The chickens showed mild retarded growth and a remarkable atrophy of the gallbladders. Microscopically, the livers showed dark brown pigments in the Kupffer cells, hepatocytes, and portal triads. These pigments showed birefringence with a Maltese-cross pattern under polarized light. Hyperplasia of the cholangioles, fibrosis, and infiltration of inflammatory cells were present in the portal triads. All the examined samples showed the same dark brown pigments in alveolar walls of the lungs. A high concentration of protoporphyrin was detected in affected livers, marrow, and feces (489, 104, and 116 μg/g wet wt., respectively) by biochemical assay.

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The immune responses of mice against glycosphingolipid (GSL) antigens and the effect of the phospholipid composition of liposomes on the immunogenicity in mice of liposome-associated GSL antigens were examined. The immunization with GSL antigen alone was unable to induce any detectable anti-GSL antibody responses. On the other hand, the immune responses against GSL antigens were detected after immunization with liposomes composed of dipalmitoylphosphatidylcholine (DPPC) (0.5 μmol), cholesterol (Chol) (0.5 μmol), Salmonella minnesota R595 lipopolysaccharides (LPS) (10 μg) and GSL (0.05 μmol) (DPPC-liposome). However, the administration with liposome composed of dimyristoylphosphatidylcholine (DMPC) (0.5 μmol), Chol (0.5 μmol), S. minnesota R595 LPS (10 μg) and GSL (0.05 μmol) and with liposomes composed of distearylphosphatidylcholine (DSPC) (0.5 μmol), Chol (0.5 μmol), and S. minnesota R595 LPS (10 μg) and GSL (0.05 μmol) was ineffective for the induction of the immune responses against GSL antigens. These results suggest that DPPC-liposome would serve effectively as a delivery vehicle for inducing immune responses against GSL antigen.

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Pulsed tissue Doppler imaging (pulsed TDI) has been demonstrated to be useful for the estimation of left ventricular (LV) systolic and diastolic functions in various human cardiac diseases. The objectives of this study were to investigate the relationship between pulsed TDI and LV function by using cardiac catheterization in healthy dogs and to evaluate the clinical usefulness of pulsed TDI in dogs with spontaneous mitral regurgitation (MR). The peak early diastolic velocity (E'), peak atrial systolic velocity (A'), and peak systolic velocity (S') were detectable in the velocity profiles of the mitral annulus in all the dogs. In the healthy dogs, S' and E' were correlated with LV peak +dP/dt and -dP/dt, respectively. E' was lower in dogs with MR than in dogs without cardiac diseases. E/E' in the MR dogs with decompensated heart failure was significantly increased in comparison with those with compensated heart failure. The sensitivity and specificity of the E/E' cutoff value of 13.0 for identifying decompensated heart failure were 80% and 83%, respectively. In addition, E/E' was significantly correlated with the ratio of left atrial to aortic diameter. These findings suggest that canine pulsed TDI can be applied clinically for estimation of cardiac function and detection of cardiac decompensation and left atrial volume overload in dogs with MR.

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The antifungal activity of β-thujaplicin was evaluated against 51 Malassezia pachydermatis strains isolated from canine ear canals with or without otitis externa. For comparison, sensitivity tests were performed on M. pachydermatis isolates for nystatin, ketoconazole, and terbinafine HCl, all clinically available antifungal agents. The minimal inhibition concentrations over 50% of the tested isolates (MIC₅₀) were 3.13 μg/ml for β-thujaplicin and nystatin, 0.016 μg/ml for ketoconazole, and 1.56 μg/ml for terbinafine HCl. The antifungal effect for M. pachydermatis of β-thujaplicin compared favorably with commercial antifungal agents. None of the 51 M. pachydermatis isolates showed resistance against any of the tested antibiotics investigated in this study. Ten representative isolates of M. pachydermatis were subcultured for 30 generations at concentrations close to the MIC levels of β-thujaplicin, nystatin, ketoconazole, and terbinafine HCl, and examined to determine whether they had acquired resistance to each drug. As a result, M. pachydermatis was found to achieve resistance more easily for ketoconazole and terbinafine HCl than for β-thujaplicin or nystatin. The MIC₅₀ of β-thujaplicin did not change during the course of subculture, and it is thought that the potential development of a resistant strain is low, even with continuous infusion for otitis externa therapy. β-Thujaplicin is an inexpensive and safe treatment with anti-inflammatory and deodorant effects that can be recommended as an effective remedy for canine otitis externa.

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Transforming growth factor-β1 (TGF-β1), an inflammatory cytokine, plays a role in tissue fibrosis, such as glomerular sclerosis and tubulointerstitial fibrosis of the kidneys. In the present study, the urinary TGF-β1 level of cats diagnosed with chronic renal failure (CRF) was measured to investigate its relationship to the pathogenesis of feline CRF. Urinary TGF-β1 levels (TGF-β1/creatinine ratio) were significantly increased compared with healthy controls, whereas serum levels of TGF-β1 were not. These results indicate that TGF-β1 is expressed in the kidneys of CRF cats, and that it was reflected in the urinary TGF-β1 level. Therefore, TGF-β1 may play a role in feline CRF, and urinary TGF-β1 could be used as a clinical marker for renal fibrosis.

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A cDNA encoding canine mucosal addressin cell adhesion molecule-1 (MAdCAM-1) was cloned. The entire open reading frame of canine MAdCAM-1 cDNA comprises 1137 bp, corresponding to 378 amino acid residues. The deduced amino acid sequence of canine MAdCAM-1 was 55.2%, 53.7%, and 52.4% identical to rat, mouse, and human MAdCAM-1, respectively. Canine MAdCAM-1 appeared to contain two immunoglobulin-like domains at the N-terminus, followed by a mucin-like domain and a third immunoglobulin-like domain. The structures of the dog, rat, and mouse proteins are likely similar because all of the cysteine residues in the immunoglobulin-like domains were conserved. Canine MAdCAM-1 mRNA was confirmed to express extremely in the mesenteric lymph node by RT-PCR.

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We conducted combined electrophysiological examinations including F-wave, motor nerve conduction velocity (MNCV), spinal cord-evoked potential (SCEP), and needle electromyography (EMG) in two cats involved in traffic accidents that consequently developed hind limb paralysis caused by lumbar hematomyelia. F-wave could no longer be elicited within 3 days after the accident, and the MNCV and compound muscle action potential (CMAP) amplitude decreased in a time-dependent manner, with CMAP no longer being evoked after 7 or 8 days. EMG showed abnormalities such as fibrillation and positive sharp waves after 6 to 8 days. These results suggest that such combined electrophysiological examinations may provide objective, quantitative data for motor nerve dysfunction in cats with lumbar hematomyelia.

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To evaluate the nutriture of Japanese black cattle with growth retardation, a metabolic profile test was carried out in 8 cattle with growth retardation and in 10 cattle with normal growth. During our observation for 1 month before blood sampling, the cattle with growth retardation ingested their forage completely. They showed lower low-density lipoprotein and albumin concentrations, and higher urea nitrogen, actoacetic acid and β-hydroxybutyric acid concentrations than the control. There were no significant differences in glucose, free fatty acid, total cholesterol, ammonia, inorganic phosphorus, and calcium concentrations between the cattle with growth retardation and the control. These data suggested that the cattle with growth retardation subjected to energy-negative condition in spite of their good forage intake.

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Acanthocephalan everted cystacanths were detected in the intestines of 61 out of 555 feral raccoons (Procyon lotor) trapped between May 2003 and April 2005 in the western region of mainland Japan (Honshu). All collected specimens were identified as the species of birds including 3 Centrorhynchus species (C. bazaleticus, C. elongatus, and C. teres), Sphaerirostris lanceoides, Plagiorhynchus ogatai, Porrorchis oti, and Southwelina hispida. Three species, C. bazaleticus, C. teres, and S. lanceoides, were new to distribution records in Japan and the Far East. Recovery rates of these acanthocephalans were higher from the raccoons trapped in spring and early summer (range 8.3-36.8%, average 21.5%) than during the remainder of the year (range 0-20.0%, average 6.3%). The reason for this seasonal change is unknown. There is little information on the acanthocephalan fauna of wild birds in Japan, however, recovery of the cystacanths from the feral raccoons captured through the year might provide useful information on the fauna and geographical distribution of avian acanthocephalans.

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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.

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DNA fragments of `Candidatus Mycoplasma haemominutum', a feline heamobartonella pathogen, were detected from unfed Ixodes ovatus collected from vegetation in Hokkaido, Fukushima and Yamaguchi Prefectures, and unfed Haemaphysalis flava in Yamaguchi Prefecture. This finding suggests that ixodid tick is a possible vector of `C. Mycoplasma haemominutum'. Spiroplasma DNA was also detected from unfed I. ovatus in Hokkaido, Fukushima and Yamaguchi Prefectures. The analysis of nucleotides sequence suggested that this Spiroplasma was distinct from registered species.

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A survey on the presence of Cryptosporidium oocyst and Giardia cyst in livestock drinking water as well as the urban tap water throughout Taiwan was carried out. Water examination for the presence of the protozoa was conducted by filtering through a PTFE membrane followed by immunomagnetic separation (IMS) and immunostaining the sediment with commercially available monoclonal antibody against Cryptosporidium and Giardia. Of the 55 different water samples from various sources examined, 2 were found to contain both of Cryptosporidium oocyst and Giardia cyst, 1 was found to contain Cryptosporidium oocyst only. These protozoa-positive water samples, originating from underground well and from the mountain spring, were also used as drinking water for livestock. However, no Cryptosporidium oocyst was found in the city tap water. This is the first report of Cryptosporidium oocyst and Giardia cyst being found in the drinking water for livestock.

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Sheep were inoculated with high tax coded pBLV-IF (H group, Nos.1-5) of bovine leukemia virus (BLV), wild tax coded pBLV-IF (W group, Nos. 6-11), or control plasmid (C group, Nos. 12-14). During the observation period (4 to 46 months), 5 of 5 cases in H group and 3 of 6 cases (Nos. 6, 7, 9) in W group became positive for gp 51. Only 1 case in H group became leukemic, and one case each of H and W groups developed lymphoma. In No. 3, lesions were found in multiple organs including the lymph nodes, gastrointestinal tract following abomasum, and heart. In No. 6, lesions of lymphoma were found only in the jejunum and heart. Morphologically, small to middle-sized lymphocytic neoplastic (NP) cells were found in both cases, but lymphoblastic NP cells were found only in No. 3. By immunohistochemical examination, the phenotypes of NP cells were determined as CD1^-, CD4^-, CD5^--, CD8α^-, sIgM⁺, λ light chain⁺, B-B4⁺, MHC class II⁺ in both case. The results of this study indicate that inoculation of pBLV-IF can induce lymphocytic and lymphoblastic leukemia/lymphoma in sheep. Additionally, it is suggested that the expression rate of tax gene is not associated with the development of leukemia/lymphoma in sheep experimentally inoculated with pBLV-IF.

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Eight new feline mammary adenocarcinoma cell lines derived from either primary or metastatic lesions were established. The morphology of all the cell lines was epithelioid and round to spindle in shape, with cell growth occurring in a monolayer fashion. On immunohistochemistry, these cells reacted with anti-keratin and anti-vimentin antisera. The doubling time of these cells was between 19 and 54 hr. Tumor masses were developed in nude mice by subcutaneous inoculation of the cells that were histologically identical to their original mammary tumor lesions. Telomerase activities measured using the telomeric repeat amplification protocol assay revealed high telemetric activity in all of the cells.

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The effect of acupuncture on intraocular pressure (IOP) was evaluated in normal dogs. After determination of baseline pressure, acupuncture was applied at 3 acupoints (LI-4, LIV-3 and GB-37) for 20 min. After acupuncture treatment, IOP were significantly lowered 2.7 ± 0.1 in left eye, 1.7 ± 0.7 in right eye, respectively (p<0.05). From these results of this study, an acupuncture therapy may be valuable treatment for decreasing on IOP in dogs.

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This study was performed to evaluate the efficacy of acupuncture on induced chronic arthritis of the dog by thermography. Complete Freund's adjuvant was injected into the left knee joint of 8 dogs to induce arthritis. Acupuncture was applied to BL-40, GB-33, GB-34, and LIV-8 once a week for 4 consecutive weeks, from 3 weeks after induction of chronic arthritis, in treatment group. At 3 weeks of acupuncture treatment, skin temperature difference (ΔT) of treatment group returned to normal range (< 0.3°C), while ΔT remained high in non-treatment group. Infrared thermography (IRT) is useful to evaluate the treatment of acupuncture for induced canine chronic arthritis. Therefore, it is considered that clinical application of IRT in arthritis treatment would be also valuable.

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Three male Beagles with low plasma luteinizing hormone and testosterone levels and spermatogenic dysfunction (SD-dogs) were given 3 weekly subcutaneous injections of a gonadotropin-releasing hormone analogue (Buserelin; GnRH-AB). The plasma T level of all of the SD-dogs increased and peaked at 4-6 weeks after the first injection. The total number of sperm and the sperm motility of the three SD-dogs increased 5-7 weeks after the first injection. Therefore, the authors concluded that 3 weekly injections of GnRH-AB transiently improves semen quality in some cases of canine spermatogenic dysfunction.

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Three individual peptide sequences, EVSHPKVG, WVTTSNQW, and SGGSNRSP, which have potentials to bind to Newcastle disease virus (NDV), were identified by the biopanning method using phage display technology. The binding specificities of these peptides presented on phages were confirmed by ELISA competition assay using chicken anti-NDV antiserum. The synthetic peptides designed based on these results partially neutralized the infection of NDV in vitro. The peptide-motives identified here have the potential to lead to the identification of novel molecules that inhibit the NDV infection independent of the immune system.

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