Yoshihisa Naito

Genomic characterization of TT viruses (TTVs) in pigs, cats and dogs and their relatedness with species-specific TTVs in primates and tupaias.

Using PCR with primers derived from a non-coding region of the human TT virus (TTV) genome, the TTV sequence in serum samples obtained from pigs (Sus domesticus), dogs (Canis familiaris) and cats (Felis catus) was identified and the entire genomic sequence was determined for each representative isolate. Three TTV isolates (Sd-TTV31 from a pig, Cf-TTV10 from a dog and Fc-TTV4 from a cat) comprising 2878, 2797 and 2064 nucleotides, respectively, each had three open reading frames (ORFs) encoding 436-635 (ORF1), 73-105 (ORF2) and 224-243 (ORF3) aa but lacked ORF4, similar to tupaia TTV. ORF3 was presumed to arise from a splicing of TTV mRNA, similar to human prototype TTV. Although the nucleotide sequence of Sd-TTV31, Cf-TTV10 and Fc-TTV4 differed by more than 50% from each other and from previously reported TTVs of 3.4-3.9 kb and TTV-like mini viruses (TLMVs) of 2.8-3.0 kb isolated from humans and non-human primates as well as tupaia TTVs of 2.2 kb, they resembled known TTVs and TLMVs with regard to genomic organization and presumed transcriptional profile rather than animal circoviruses of 1.7-2.3 kb. Phylogenetic analysis revealed that Sd-TTV31, Cf-TTV10 and Fc-TTV4 were closer to TTVs from lower-order primates and tupaias than to TTVs from higher-order primates and TLMVs. These results indicate that domestic pigs, cats and dogs are naturally infected with species-specific TTVs with small genomic size and suggest a wide distribution of TTVs with extremely divergent genomic sequence and length in animals.

Comparative Histology of the Laminar Bone between Young Calves and Foals

Laminar bone or primary plexiform tissue, not Haversian bone, shows an alternative concentric pattern of laminar-bone units or plates around the bone marrow periphery of long bones, although the laminar bone is gradually replaced by osteons during the growth period. One laminar-bone unit is constructed with a hypercalcified line in the center, woven bone on both sides of the line, and lamellar bone with laminated appositional lines. Such a laminar bone showing a homogeneous calcification has been reported in young calves and some young large animals, but it has not been reported in foals although a previous report proposed that the bone structure was distinguishable from plexiform tissue. In this study, we compared young calves with foals by backscattered electron imaging mainly of transverse ground sections of mid-diaphysis. Foals had many hypercalcified lines arranged concentrically around the bone marrow periphery, which were similar to those of young calves. However, rows of cylindrical osteon-like structures with Haversian canal-like canals running along the long-bone axis were arranged between the concentric hypercalcified lines. Each Haversian canal-like structure was enclosed with laminated appositional rings of lamellar bone deposited on the woven bone. In the developing period, the bone units containing the concentric hypercalcified lines were basically equal to the laminar-bone units. The osteon-like structures or ‘pseudo-osteons’ were gradually replaced by ‘true osteons’ during the growth period. The blood vessels in the Haversian canal-like canals of foals ran along the long-bone axis, whereas the blood vessels in the concentrically prolonged bone cavities of young calves ran transversely to obliquely against the long-bone axis. Thus, the long-bone cortex of foals showing an alternative concentric pattern of a row of the osteon-like structures arranged between the hypercalcified lines will be histologically classified into a variety of laminar bone caused by the different arrangement of blood vessels. Such a laminar bone may have a biomechanical structure against physical stress, especially the modified laminar bone of foals with osteon-like structures, when compared with the typical concentric laminar bone of young calves and also Haversian bone possessing variously calcified numerous osteons caused by bone remodeling.

The evaluation of the curd forming ability of milk replacers

Inadequate milk curd formation in the abomasum of newborn calves causes malnutrition and diarrhea. In order to define the factors of inadequate curd formation, we compared the curd forming ability among 9 kinds of milk replacers, bulk milk (raw milk), and skim milk both in vitro and in vivo. When rennet was added, the raw milk and one milk replacer formed firm curds. The rest of the milk replacers and skim milk did not form any curd. When a solution of HCl was added, raw milk, three milk replacers and skim milk formed the curd at pH 4.5, but the other milk replacers did not. When HCl was added following the rennet, raw milk, one milk replacer and skim milk formed the curd. In vivo, raw milk, two milk replacers and skim milk showed good curd formation whereas the other milk replacers showed poor curd formation inside the abomasums of the calves. This study showed that most of the milk replacers sold in Japan could not form the curd with rennet.

Molecular Screening of Canine GM1 Gangliosidosis Using Blood Smear Specimens after Prolonged Storage: Detection of Carriers among Shiba Dogs in Northern Japan

Molecular screening of GM1 gangliosidosis in Shiba dogs was carried out in northern Japan using blood smear specimens after prolonged storage. Of 125 specimens obtained from 3 veterinary teaching hospitals for this screening, 68 specimens (54%) were adequate for direct amplification in a polymerase chain reaction (PCR)-based DNA test, and the percentage of adequacy was different at each hospital (34%, 73%. and 100%), suggesting that the amount of blood on the smear and the storage condition of specimens may affect adequacy. Of the 68 dogs examined, 2 dogs (2.9%) were heterozygous carriers for this disease and the other dogs were all genotypically normal. The results suggest blood smear specimens can be useful for PCR testing after prolonged storage provided specimens contain a generous amount of blood and have been adequately stored. The study also suggests that GM1 gangliosidosis may be widely prevalent in the Shiba dog population in northern Japan.

A Case of Japanese Black Steer Characterized by Epileptic Seizures

pupillary and corneal reflex. Symptomatic epilepsia continuing for 11 to 18 hours was recognized seven times at intervals of 15 hours to 6 days during the observation period of 28 days. After these syndromes disappeared, the steer got up and recovered normal vigor and appetite, but was gradually emaciated thereafter and fell into sopor. Electroencephalographically, a spectrum resembling the deep sleep wave was obtained in spite of the activated state of the brain. That is, ƒÀ-wave was extremely rare and ƒÂ-wave overwhelming in the spectrum. No stimulations by flash or sound changed the patterns of the spectrum. Histo-

Mycotic bronchitis in a dog affected with distemper.

体重9kg, 2歳の雌柴犬で, 発熱, 食欲不振, 下痢, 嘔吐を主徴とし, 多量の抗生物質その他の治療を受けたが改善を見ず, 発病後21日目に安楽死を行なった. 殺後の組織学的検査により, 強いジステンパー病巣と共に, 左肺横隔膜葉の気管枝内に, 形態学的にAspergillus sp.と考えられる菌糸塊をいれ, これから伸びる菌糸が粘膜に侵入し, 気管枝壁の壊死と強い細胞性惨出を伴う, 犬ではきわめて稀とされる真菌性気管枝炎が認められた.