Pedigree analysis of factor XI deficiency in Japanese black cattle

Using a DNA-based diagnostic test for factor XI deficiency in Japanese black cattle, we surveyed 123 cattle (42 sires and 81 dams) in Gifu and Hyogo prefectures, and calculated gene frequencies. In sires, we drew up the pedigree network of the cattle with the factor XI deficiency. Results showed that the mutated allele of factor XI deficiency was retroactive in at least 6 or more generations of sires. Frequencies of the mutant gene were higher at 26.4% in total, and at 33.3% in sires. All 7 cattle with the homozygote of mutated allele were clinically normal, and showed no bleeding episodes. The mutated allele of factor XI deficiency might be widespread among Japanese black cattle.     

Biological roles of monoterpene volatiles derived from rough lemon (<Emphasis Type="Italic">Citrus jambhiri</Emphasis> Lush) in citrus defense

Volatile compounds of plants, including monoterpenes, are a possible source of signal molecules that induce defense systems to protect plants from tissue damage. Volatile compounds from rough lemon leaves were trapped by solid-phase microextraction fibers in sealed vials, and subsequent gas chromatography–mass spectrometry and gas chromatography analyses identified the profile of the major components, mainly various monoterpenes. Among several monoterpenes examined, citral, citronellal, and linalool significantly inhibited the spore germination and hyphal growth of Alternaria alternata. The effect of linalool was fungistatic, while the effects of citral and citronellal were partially fungicidal. Wounding of rough lemon leaves induced a significant increase in release of monoterpenes. The release of linalool was the most abundant and was 14.5 times that of unwounded rough lemon leaves. Unlike the wounding treatment, microbe attack did not significantly change monoterpene releases, and there was statistically no difference found in the peak areas from microbe-treated and untreated leaves. Linalool, limonene, and β-pinene also had insect-repellant effects on wild-type Drosophila melanogaster. Expression patterns of defense-related genes in rough lemon and rice significantly changed after treatment with vapors of monoterpene volatiles. Taking these results together, monoterpene volatiles are likely to play roles in the defense of rough lemon against microbe and insect pathogens.     

Characterization of white sapote (Casimiroa edulis Llave & Lex.) germplasm using floral morphology,RAPD and AFLP markers

Floral morphology, random amplified polymorphic DNA (RAPD), and amplified fragment length polymorphism (AFLP) were used to characterize and verify genetic diversity within a white sapote cultivar collection and to develop molecular markers for germplasm identification. On the basis of floral morphology, the cultivars were classified into three types: type I included 23 cultivars with large ovaries and small anthers; type II included 13 cultivars with small ovaries and large anthers; and type III included one cultivar, named ‘Maltby’, with a large ovary and large anthers. DNA was isolated from 39 cultivars of white sapote and subjected to RAPD and AFLP analysis using 24 and 7 primers, respectively. One hundred and sixty-eight RAPD and 286 AFLP bands were used to assess genetic characterization among white sapote. Sixty percent of the RAPD and 77% of the AFLP amplification products were polymorphic among accessions. RAPD or AFLP markers differentiated all white sapote cultivars effectively. Moreover, each flower type was characterized as specially associated with two RAPD bands. UPGMA dendrograms based on RAPD and AFLP data, showed the majority of the cultivars from flower type I and flower type II clustering together. Finally 101 RAPD markers and 220 AFLP markers were used to construct a neighbor-joining dendrogram. This showed that the 37 cultivars could be classified into six distinct clusters, between which the similarity coefficient was as low as 0.00–0.55, even though the cultivars were morphologically very similar. The remaining two cultivars namely ‘Smathers’ and ‘Maltby’ were found genetically very distant from the other cultivars in RAPD, AFLP or combined RAPD and AFLP based dendrograms. The results suggested that the level of genetic variation among white sapote cultivars is diverse and the morphological and molecular data may lead to representation of the cultivar relationships as well as flower type discrimination.     

Successive changes of hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta).

Hematologic characteristics and plasma chemistry values of juvenile loggerhead turtles (Caretta caretta) from the ages of 1 mo to 3 yr were obtained to establish baseline values. Five clinically normal loggerhead turtles were selected from the same clutch and raised in an indoor artificial nesting beach. Blood samples were successively collected and examined for various blood characteristics for a maximum total of 15 times. Hematologic characteristics, including packed cell volume, white blood cell counts, and white blood cell differentials; and plasma chemistry values, including total bilirubin, total protein, albumin, glutamic oxaloacetic transaminase, glutamic pyruvic transaminase, gamma-glutamic transpeptidase, creatinine, blood urea nitrogen, uric acid, alkaline phosphatase, amylase, triglyceride, total cholesterol, ionized sodium, ionized potassium and ionized chlorine, were measured. These results were used to establish a hematology and blood chemistry baseline for captive juvenile loggerhead turtles and will aid in their medical management.     

Digestive enzyme activities of pancreas and intestinal digesta in streptozotocin-induced diabetic piglets

Six (Landrace × Large White) × Duroc crossbred, castrated weanling piglets were used to evaluate the effect of a streptozotocin (STZ) injection to induce diabetes on the activities of digestive enzymes derived from the pancreas (lipase, amylase, trypsin and chymotrypsin) in the pancreas and intestinal digesta. Fasting plasma glucose after the administration of STZ was maintained at the level of 302–491 mg/dL during this experiment, compared with the level of 89–125 mg/dL in the control group, and they were defined as the STZ‐induced diabetic piglets for about 7 weeks. Although their bodyweight increased in proportion to a quadratic curve (P < 0.0001) during 49 days after the administration of STZ, the growth of the STZ‐induced diabetic piglets was slower compared with the control. The STZ‐injection did not affect the percentage of the pancreas in bodyweight. The administration of STZ in the piglets tended to accelerate the activity of lipase (P = 0.06) and depress the activity of amylase (P = 0.15) or chymotrypsin (P = 0.18), as units/kg bodyweight, in the pancreas. In the case of measurement as units/kg bodyweight, the activities of intestinal digesta in the STZ group showed a tendency to be higher than those in control group, irrespective of the sort of enzymes. In conclusion, STZ‐induced diabetic piglets have a moderate digestive ability and the administration of exogenous digestive enzymes is not necessary when they are used as a diabetic animal model.     

Acinic cell carcinoma in an African pygmy hedgehog (Atelerix albiventris)

Abstract: A male African pygmy hedgehog ( Atelerix albiventris ), estimated to be 3 years old, presented with exophthalmos and fixed abduction of the right eye. Radiographic examination revealed a retrobulbar tumor in the right orbital cavity. The mass was surgically resected but recurred 3 months later and the hedgehog died. There was no gross or microscopic evidence of salivary or lacrimal gland involvement of the tumor at surgery or at necropsy. The histopathologic, immunohistochemical, and ultrastructural findings were those of acinic cell carcinoma, the origin of which was unknown. This is the first known case of acinic cell carcinoma in an African hedgehog.     

Cytochrome oxidase I gene sequence of Hypoderma sinense infecting yaks in the Qinghai-Tibet high plateau of China

The previous scanning electron microscopy study of the Hypoderma species suggested that Hypoderma sinense Pleske (H. sinense) was different from Hypoderma bovis (H. bovis) and Hypoderma lineatum (H. lineatum). In this study, the mitochondrial cytochrome oxidase I (COI) gene sequence of H. sinense was compared with those of the other two species. The H. sinense sequence showed only 88.3% homology to H. bovis and 88.5% to H. lineatum. The results of the sequencing analysis confirm that H. sinense is a different species from H. bovis and H. lineatum.     

Genetic and serological characterization of novel serotype G8 bovine group A rotavirus strains isolated in Japan

G8 bovine group A rotaviruses isolated in Japan were genetically and serologically characterized. The VP7 gene nucleotide and amino acid sequences revealed high identity with each other. All Japanese G8 strains were classified into the same lineage in the phylogenetic analysis based on VP7 gene sequences. Antisera to four Japanese G8 strains neutralized other G8 strains, but their neutralizing titers were between 8-fold lower and 2-fold higher than homologous strains. These results suggest that the VP7s of Japanese G8 strains have similar genetic and serologic characteristics. Observed differences in the neutralizing abilities of antisera for each strain appear to depend on differences in the P serotypes/genotypes.     

Assessment of bovine X- and Y-bearing spermatozoa in fractions by discontinuous percoll gradients with rapid fluorescence in situ hybridization

This study was designed to apply the method of discontinuous Percoll gradients for sex preselection in bovine semen by using a current developed molecular technique, fluorescence in situ hybridization (FISH). In addition, we attempted to amplify the level of enrichment of X- or Y-bearing spermatozoa by treating for activating sperm motility performance with 10 mM caffeine. Bovine spermatozoa were fractionated on Percoll gradients into two major subpopulations of motile spermatozoa (bottom fraction) and weak motile spermatozoa (top fraction). The percentage of Y-bearing spermatozoa in the top fraction (52.9%) slightly exceeded and that in the bottom fraction (44.3%) decreased significantly (P<0.001) compared with the theoretical ratio (50:50). Washing sperm with BO medium affected a deviation between the two sex populations, whereas semen activated with caffeine showed no difference in the percentage of X- and Y-bearing spermatozoa in both fractions compared with the theoretical ratio (50:50). These results show that the proportion of X- and Y-bearing bovine spermatozoa can deviate after discontinuous Percoll gradients, although the proportion of X- and Y-bearing bovine spermatozoa was affected by sperm motility of the sample applied.     

Immunocytochemical evaluation of epithelial–mesenchymal transition in epithelial tumors of dogs and cats

Epithelial–mesenchymal transition (EMT) plays a crucial role in metastasis of epithelial tumors; however, it is challenging to detect EMT by cytology. In the present study, EMT was visualized by fluorescence-immunocytochemistry (FICC). Air-dried smears from epithelial tumors of dogs (n=22) and cats (n=9) were stained using mouse monoclonal anti-E-cadherin and rabbit monoclonal anti‐vimentin antibodies. Enzymatic immunohistochemistry (IHC) revealed that 51.6% (8/22 in dogs, 8/9 in cats) of the cases showed EMT. In dogs, FICC could detect EMT in 62.5% (5/8) of those cases. In cats, FICC could detect EMT in 100% (8/8) of the cases. In conclusion, the present FICC method could successfully detect EMT using conventional air-dried cytology smear slides.