Serum growth hormone and insulin-like growth factor-1 concentrations in Japanese black cattle with renal tubular dysplasia

To elucidate the cause of growth retardation, 14 cattle with renal tubular dysplasia (RTD) were investigated by determining serum growth hormone (GH) and insulin-like growth factor (IGF)-1 concentrations. Compared with 6 healthy cattle, the baseline, maximum, area under the curve and peak amplitude of serum GH concentrations were higher or tended to be higher. Serum IGF-1 concentrations were lower normal or lower in cattle with RTD than in healthy cattle. Serum IGF-1 concentrations correlated significantly with body weight and chest girth, but not with serum GH and creatinine concentrations. Growth retardation in RTD might be associated partially with GH resistance.     

Clinical features of renal tubular dysplasia, a new hereditary disease in Japanese Black cattle

A new hereditary disease characterised by renal failure, poor growth and long hooves in Japanese Black cattle (wagyu) has been recognised in a region of central Japan since 1990. The number of calves affected has increased gradually, with the incidence reaching 17 of 485 (3.51 per cent) in 1995. Almost all the calves were slightly undersized at birth, and repeatedly had diarrhoea during the neonatal period. They began to show signs of growth retardation with proportional body and elongation of the hooves from about two to five months of age, but they had an almost normal or only slightly decreased appetite. The concentrations of urea nitrogen, creatinine and inorganic phosphorus in serum were high, and the affected calves excreted diluted urine frequently. Among 25 cases, the urine of 21 contained occult blood, 24 contained protein and two contained glucose. In 29 calves observed for 30 to 130 days, the course of the disease varied; in 21 of them it remained unchanged, six became gradually worse and two became severely debilitated and died. The disease was diagnosed as renal tubular dysplasia by histopathological examination.     

Multicystic renal dysplasia in a Japanese black bull.

Multicystic renal dysplasia was found in a 6-day-old Japanese black bull. Grossly, both kidneys were markedly small (2.0 x 3.5 cm) with numerous cysts ranging from 1 to 8 mm in diameter. Histopathologically, both kidneys consisted of many irregularly enlarged cysts, immature glomeruli, small ducts and anomalous stromal connective tissues containing focal persistent mesenchyme characterized by a proliferation of stellate cells with myxomatous area. These features are compatible with those of multicystic renal dysplasia in humans and other mammals.     

Renal reabsorption of magnesium and calcium by cattle with renal tubular dysplasia

The concentrations of magnesium and calcium in the serum and urine and their rates of clearance were determined in cattle with renal tubular dysplasia, an autosomal recessive hereditary disease associated with a deletion of the paracellin-1 gene in Japanese Black cattle. There were no significant differences in the serum or urine magnesium concentrations between normal cattle and cattle which were heterozygous or homozygous for the condition. Serum calcium concentrations tended to be lower in the homozygous cattle, and the serum creatinine and urea nitrogen concentrations were significantly higher in the homozygous cattle. The ratio of magnesium:creatinine and the fractional excretion of magnesium were higher in cattle with the disease than in normal cattle. There were no significant differences in urine calcium concentration, the calcium:creatinine ratio, and fractional excretion of calcium between normal cattle and cattle which were homozygous or heterozygous for the condition. The creatinine clearance was significantly lower in the homozygous cattle than in normal cattle. The clearance, excretion rate, reabsorption rate and reabsorption rate:clearance ratio of magnesium in cattle with renal tubular dysplasia were significantly lower than in normal cattle. The clearance rate and reabsorption rate of calcium were also significantly lower in the affected cattle, but the excretion rate and reabsorption rate:clearance of calcium were not different between the normal cattle and the cattle homozygous for the condition. In cattle with the condition the rate of reabsorption of magnesium by the kidneys was low, but the rate of reabsorption of calcium was normal.     

Idiopathic thrombocytopenia in Japanese black cattle

A female Japanese Black cow showed hemorrhage under the mucous membrane of the eye conjunctiva and the vagina, and thrombocytopenia in peripheral blood. The clinical manifestations repeated spontaneously during long-term observation. The clinical findings were stable except for severe periodic bleeding. Prothrombin time, activated partial thromboplastin time and fibrinogen levels were normal. The anti-platelet antibody in the serum was analyzed by indirect platelet coagulation tests. Because the platelets in the micro-plate wells of the subject series showed coagulation up to 1:128, it was judged that anti-platelet antibody was present in the serum. A number of megakaryocytes were recognized in smear samples obtained from bone marrow of the scapula immediately after euthanasia. Therefore, this case was diagnosed as idiopathic thrombocytopenia.     

Optic pathway degeneration in Japanese black cattle

Degeneration of the optic pathway has been reported in various animal species including cattle. We experienced a case of bilateral optic tract degeneration characterized by severe gliosis in a Japanese black cattle without any obvious visual defects. To evaluate the significance, pathological nature and pathogenesis of the lesions, we examined the optic pathway in 60 cattle (41 Japanese black, 13 Holstein and 6 crossbreed) with or without ocular abnormalities. None of these animals had optic canal stenosis. Degenerative changes with severe gliosis in the optic pathway, which includes the optic nerve, optic chiasm and optic tract, were only observed in 8 Japanese black cattle with or without ocular abnormalities. Furthermore, strong immunoreactivity of glial fibrillary acidic protein was observed in the retinal stratum opticum and ganglion cell layer in all 5 cattle in which the optic pathway lesions could be examined. As etiological research, we also examined whether the concentrations of vitamin A and vitamin B12 or bovine viral diarrhea virus (BVDV) infection was associated with optic pathway degeneration. However, our results suggested that the observed optic pathway degeneration was probably not caused by these factors. These facts indicate the presence of optic pathway degeneration characterized by severe gliosis that has never been reported in cattle without bilateral compressive lesions in the optic pathway or bilateral severe retinal atrophy.     

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.     

Detection of novel kobu-like viruses in Japanese black cattle in Japan

During surveillance for bovine diarrhea of unknown causes in Japanese black cattle in Kagoshima Prefecture,Japan, we found two types of novel kobu-like viruses in fecal samples of calves. Sequence analyses revealedthat they had L protein and 2A protein with H-box/NC sequence motif, which are present in kobuviruses.Phylogenetic analysis revealed that they were related to kobuviruses; however, they clustered apart from otherkobuviruses. In the prevalence study of two types of novel kobu-like viruses, 16.9% and 10.4% prevalence ofthese viruses were observed in the feces of diarrheal calves in this area.     

Economic losses related to internal diseases in Japanese black cattle

The objective of this study was to estimate the direct economic losses due to the condemnation of the liver and large intestine because of internal diseases (multifocal necrosis in the liver (MNL) and inflammation of the large intestine (ILI)), and the indirect losses because of reductions in carcass performance from MNL, bovine abdominal fat necrosis (BFN) and ILI using data from 5383 Japanese Black cattle. Direct losses were estimated by multiplying the price of the condemned part by the frequency of its occurrence owing to the disease. Similarly, indirect losses were estimated as the product of unit carcass price and reduction in carcass weight (CW) due to the disease. The direct impact on the beef cattle industry from MNL and ILI was estimated at around $1.29 million (US$1 = ¥120) per year. A least‐squares analysis showed that MNL had no influence on any carcass trait, whereas BFN and ILI significantly reduced CW, rib eye area and darkened the beef. ILI also reduced rib thickness. The indirect losses from BFN and ILI were estimated as a maximum of $131.7 and $256.4 per animal and around $6.26 million and $4.03 million for the industry, respectively, mostly because of the reduction in CW.     

Carrier rate of Factor XI deficiency in stunted Japanese black cattle

Blood examinations and genotyping of Factor XI (F11) were performed in growth retardation Japanese Black cattle and their dams. Genotyping of F11 revealed that the recessive homozygous and heterozygous genotype frequencies were 5.2% and 50.0% in the Claudin-16 (CL-16) deficiency group (n=58), 0% and 14.2% in the renal dysplasia group (n=7), 0% and 26.1% in the non-CL-16 deficiency nephritis group (n=23), 8.9% and 46.7% in the hypogenesis syndrome group (n=45), 6.2% and 25.0% in the neonatal weak calf syndrome group (n=32), 9.1% and 38.6% in the respective dams group (n=44), 0% and 23.1% in the normal cattle group (n=13), and 5.9% and 38.2% in total (n=222), respectively. These results showed that the carrier rate of F11 deficiency was high in Japanese Black cattle, and that the CL-16 deficiency, hypogenesis syndrome, neonatal weak calf syndrome, and dams groups had a large amount of recessive homozygous genotype than the other groups. No abnormal bleeding was observed clinically in the present study, and 4 of the recessive homozygous dams showed normal growth and parturition.     

Abnormal development of nephrons in claudin-16-defective Japanese black cattle

The kidneys of 37 Japanese Black calves aged 2 to 65 months diagnosed with Claudin 16 (CL-16) defect by the DNA-based test were examined pathologically. The animals exhibited clinical symptoms such as growth impairment, renal failure, overgrowth of hooves, and anemia at a young age. There was no correlation between the time of onset and age. Kidney weights relative to body weight were similar to those in normal animals, but both kidney net weights and size were reduced due to atrophy in animals that showed severe renal dysfunction. Histopathological examination of the kidneys showed reduction in the number of glomeruli, compensatory hypertrophy of glomeruli and tubules, and glomerular and tubular atrophy accompanied by interstitial fibrosis and lymphocytic infiltration. Glomeruli were clearly less in number in the kidneys of CL-16-defective animals than those of normal animals even in the cases with mild lesions. A small number of immature glomeruli and tubules were also detected, suggesting that there were fewer nephrons developed at birth in CL-16-defective animals. It was suggested that a defect of the CL-16 gene is involved in the "abnormal development of nephrons". Immunohistopathological examination of the kidneys showed that the epithelium of thick ascending limb of Henle was stained with anti-CL-16 antibody in the control animals, but not in the affected animals, suggesting a defect of CL-16 in the epithelium of renal tubules in the affected animals.     

Premature capacitation of frozen-thawed spermatozoa from subfertile Japanese black cattle

Artificial insemination (AI) subfertility is an indication of failure of AI with frozen-thawed sperm classified as normal by conventional semen examination. Recently, 8 AI-subfertile Japanese Black cattle (S1-S8) were identified using the routine AI test or in vivo fertilization test, which included AI with frozen-thawed sperm of superovulated females and subsequent non-surgical recovery of presumptive zygotes. In the present study, we assessed capacitation states and in vitro oocyte penetration of frozen-thawed sperm from these bulls to estimate causal factors of AI subfertility. Frozen-thawed sperm from 8 AI-subfertile (S1-S8) and 9 fertile (F1-F9, control) bulls were washed and then used for a chlortetracycline (CTC) staining assay and in vitro fertilization test. The CTC staining assay revealed that approximately 50% of the sperm from 4 of the AI-subfertile bulls (S5-S8) were prematurely progressing into the capacitation state immediately after washing and resuspension in a CaCl(2)-lacking medium. In contrast, most of the sperm from the fertile bulls and other AI-subfertile bulls (S1-S4) remained uncapacitated. Addition of CaCl(2) to the medium effectively promoted a spontaneous acrosome reaction in the sperm samples from the AI-subfertile bulls (S5-S8). Moreover, the in vitro fertilization test showed that rates of sperm penetration into oocytes were significantly lower in sperm samples from the AI-subfertile bulls (S5-S8) than in the control sperm samples from the fertile bulls (F2-F4 and F7-F9). It has previously been suggested that prematurely capacitated sperm undergo a spontaneous acrosome reaction possibly due to uncontrolled influx of calcium ion, and consequently they possess relatively lower in vitro fertilizing ability. It is therefore possible that premature capacitation of sperm used for AI is a causal factor of subfertility of male Japanese Black cattle and a potentially good marker for identification of subfertile bulls for removal from AI programs.     

Results of metabolic profile test in Japanese black cattle with growth retardation

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 beta-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.     

Endocrine patterns in two strains of Japanese black cattle with growth retardation

Endocrine patterns were compared in 2 strains of Japanese black cattle with growth retardation; MHO- and HSK-paternal strains (MHO and HSK cattle, respectively). MHO cattle (n=8) displayed lower serum concentrations of insulin-like growth factor-1 (IGF-1), triiodothyronine (T3), thyroxine (T4), and cortisol (31.1+/-20.7 ng/ml, 73.9+/-51.9 ng/dl, and 2.9+/-2.9 microg/dl, 1.3+/-0.7 microg/dl, respectively) than those in both HSK cattle (n=5) (64.9+/-47.6 ng/ml, 97.8+/-40.7 ng/dl, 4.1+/-2.1 microg/dl and 1.8+/-1.1 microg/dl, respectively), and the controls (n=6) (314.7+/-197.2 ng/ml, 140.2+/-21.3 ng/dl, 5.8+/-1.7 microg/dl, and 3.0+/-1.4 microg/dl, respectively). The area under the concentration curve of growth hormone (GH-AUC 0-600 min) in MHO cattle (22210+/-18951 ng.min/ml) tended to be greater than those in HSK (7887+/-6340 ng.min/ml) and the controls (2811+/-1275 ng.min/ml). MHO cattle showed a high GH-AUC0-600 min in contrast to a low serum IGF-1 concentration, as well as lower serum T3, T4, and cortisol concentrations. HSK cattle exhibited the same secretory patterns, but much more moderately. Growth retardation in Japanese black cattle exhibits some variations based on pedigree.     

Structural changes in intramuscular connective tissue during the fattening of Japanese black cattle: effect of marbling on beef tenderization

We investigated changes in structures and mechanical properties of the intramuscular connective tissue during the fattening of Japanese Black steers, using the cell maceration method for scanning electron microscopy. During the early fattening period, from 9 to 20 mo of age, collagen fibrils of the endomysium in longissimus muscle associated more closely with each other, and collagen fibers in the perimysium increased in thickness and their wavy pattern became more regular. These changes were closely related to the increase in mechanical strength of the intramuscular connective tissue and resulted in a toughening of the beef during the period. The shear force value of longissimus muscle decreased after 20 mo of age, concomitantly with the rapid increase in the crude fat content. Scanning electron micrographs of the longissimus muscle dissected from 32-mo-old steers clearly showed that the adipose tissues were formed between muscle fiber bundles, that the honeycomb structure of endomysia was partially broken, and that the perimysium separated into thinner collagen fibers. In semitendinosus muscle, in which the crude fat content was lower (P<.05) than that in longissimus muscle, the structure of the intramuscular connective tissue remained rigid at 32 mo of age. The shear force value of the muscle increased even in the late fattening period, from 20 to 32 mo of age. Thus, the development of adipose tissues in longissimus muscle appears to disorganize the structure of the intramuscular connective tissue and contributes to tenderization of highly marbled beef from Japanese Black cattle during the late fattening period.     

Prevalence of Borna disease virus antibodies in healthy Japanese black cattle in Kyushu

Epidemiological studies have demonstrated that asymptomatic infection of Borna disease virus (BDV) is found in various species of animals in Japan. Recent reports have also revealed that neurological diseases caused by this virus could exist in horses, cattle, a dog, and cats in this country. In this study, we investigated seroprevalence of BDV antibodies in Japanese black cows reared in Kyushu, the southernmost main island of Japan, using ELISA and Western-immunoblotting. Of 101 serum samples, 11 (10.9%) and 21(20.7%) sera were identified as having antibodies to the BDV N and P antigens, respectively. Among the positive sera, three cows (2.9%) were seropositive for both of the antigens. Furthermore, interestingly, only female cows showed antibodies to P, whereas N antibodies were detected in male and female cows with a comparative ratio. Together with previous studies, our results indicate that BDV might be widely spread in cattle raised in Japan. Furthermore, this is the first report to show that beef cattle, Japanese black cattle, have antibodies against a possible zoonotic pathogen, BDV.     

Gene dropping analysis of founder contributions in a closed Japanese black cattle population

Gene dropping simulation was applied to Japanese Black cattle population in Hyogo prefecture, to examine the survivals of alleles originated from founder animals. In the analysis, unique alleles were assigned to founders, and the genotypes of all descendants along the actual pedigree were generated through Monte Carlo simulation following Mendelian segregation rules. By replicating this process 10 000 times, the distribution of frequencies of alleles from each founder was estimated. From the distribution, several quantities useful for the management of genetic diversity, such as the probability of allele extinction and the probability of alleles surviving at a critically low frequency were derived. The materials used were 68 781 animals born in 1955–1998 and their pedigree records traced back to the population in 1937 or before. The expected number of alleles retained in the population drastically decreased during the analyzed period, and reached to 57.9 in the population of 1998, which was only 3.3% of the total number of alleles assigned to founders. Detailed analysis of major founders with relatively high genetic contributions to the current population revealed that alleles from most of the major founders are now at high risk of future extinction. These results strongly suggest that for the management of genetic diversity, the genetic contributions of founders are not fully informative, and emphasize the importance of the detection of live animals having founder alleles with high extinction possibilities.