Oversight probability in presence–absence sampling for detection of fish eggs

ABSTRACT:   Presence–absence sampling (PAS) is a sampling technique that observes only the presence or absence of objects, such as eggs, in sampling units. The purpose of the present paper was to study the oversight probability in a PAS study for detecting fish eggs in connection with the spatial distribution of eggs. It was shown that the oversight probability of presence of eggs under no-eggs observation is a monotone decreasing function of the parameter of the degree of contagion.     

Influence of low and high temperatures on the initiation and the development of a bulb primordium in isolated tulip embryos

The influence of 4°C and 24°C on the germination and the subsequent development of isolated tulip embryos was studied, using embryo culture techniques. A low temperature not only influenced the breaking of the embryo dormancy, but also the initiation of a bulb primordium. High-temperature treatment after chilling accelerated the development of an initiated bulb primordium.     

The relationship between plasma leptin concentrations and the distribution of body fat in crossbred steers

The objective of the present study was to examine the relationship between plasma leptin levels and fat distribution in crossbred steers. Twenty-four crossbred steers (Japanese Black × Holstein) were used in this study. Six steers belonged to the growing phase (11 months of age), and 18 steers to the finishing phase (from 21 to 28 months of age). Plasma leptin levels correlated positively with body fat weight ( r  = 0.807, P  < 0.01), body fat percentage ( r  = 0.802, P  < 0.01), visceral fat weight ( r  = 0.807, P  < 0.01), visceral fat percentage ( r  = 0.780, P  < 0.01), subcutaneous fat weight ( r  = 0.745, P  < 0.01), subcutaneous fat percentage ( r  = 0.734, P  < 0.01), intermuscular fat weight ( r  = 0.793, P  < 0.01), intermuscular fat percentage (0.775, P  < 0.01), renal fat weight ( r  = 0.694, P  < 0.01), renal fat percentage ( r  = 0.642, P  < 0.01), subcutaneous fat thickness ( r  = 0.755, P  < 0.01), intermuscular fat thickness ( r  = 0.721, P  < 0.01), beef marbling score ( r  = 0.706, P  < 0.01) and the crude fat content of Musculus longissimus thoracis ( r  = 0.592, P  < 0.01). The location of fat deposits did not affect the correlation values. However, when steers were divided into growing ( n  = 6) and finishing ( n  = 18) groups, there was no significant correlation between plasma leptin levels and the carcass characteristics related to fat deposition (fat weight, fat percentage, carcass traits) in either group. These results suggest that plasma leptin levels in crossbred steers positively correlate with body fat content and carcass traits when these parameters vary widely.     

Sexing of Dog Sperm by Fluorescence In Situ Hybridization

Effective preselection of sex has been accomplished in several species of livestock and also in humans using the flow cytometric sperm sorting method. A guaranteed high sorting accuracy is a key prerequisite for the widespread use of sperm sexing. The standard validation method is flow cytometric remeasurement of the DNA content of the sexed sperm. Since this method relies on the same instrument that produced the original sperm separation, it is not truly independent. Therefore, to be able to specifically produce either male or female offspring in the dog, we developed a method of direct visualization of sex chromosomes in a single sperm using fluorescence in situ hybridization (FISH) as a validation method. Denaturation of canine spermatozoa by immersion in 1 M NaOH for 4 min yielded consistent hybridization results with over 97% hybridization efficiency and a good preservation of sperm morphology. There was no significant difference between the theoretical ratio (50:50) and the observed ratio of X- and Y-chromosome-bearing spermatozoa in any of the three dogs. In addition, the mean purities of flow-sorted sex chromosomes in spermatozoa of the three dogs were 90.8% for the X chromosome fraction and 89.6% for the Y chromosome fraction. This sorting was evaluated by using the dual color FISH protocol. Therefore, our results demonstrated that the FISH protocol worked reliably for both unsorted and sexed sperm samples.     

Dynorphin-Kappa Opioid Receptor Signaling Partly Mediates Estrogen Negative Feedback Effect on LH Pulses in Female Rats

Accumulating evidence suggests that the arcuate nucleus (ARC) kisspeptin/neurokinin B (NKB)/dynorphin (KNDy) neurons play a role in estrogen negative feedback action on pulsatile gonadotropin-releasing hormone (GnRH)/luteinizing hormone (LH) release. The present study aimed to determine if dynorphin (Dyn) is involved in estrogen negative feedback on pulsatile GnRH/LH release. The effect of the injection of nor-binaltorphimine (nor-BNI), a kappa-opioid receptor (KOR) antagonist, into the third cerebroventricle (3V) on LH pulses was determined in ovariectomized (OVX) adult female rats with/without replacement of negative feedback levels of estradiol (low E₂). The mean LH concentrations and baseline levels of LH secretion in nor-BNI-injected, low E₂-treated rats were significantly higher compared with vehicle-treated controls. On the other hand, the nor-BNI treatment failed to affect any LH pulse parameters in OVX rats without low E₂ treatment. These results suggest that Dyn is involved in the estrogen negative feedback regulation of pulsatile GnRH/LH release. The low E₂ treatment had no significant effect on the numbers of ARC Pdyn (Dyn gene)-,Kiss1- and Tac2 (NKB gene)-expressing cells. The treatment also did not affect mRNA levels of Pdyn and Oprk1 (KOR gene) in the ARC-median eminence region, but significantly increased the ARC kisspeptin immunoreactivity. These findings suggest that the negative feedback level of estrogen suppresses kisspeptin release from the ARC KNDy neurons through an unknown mechanism without affecting the Dyn and KOR expressions in the ARC. Taken together, the present result suggests that Dyn-KOR signaling is a part of estrogen negative feedback action on GnRH/LH pulses by reducing the kisspeptin release in female rats.     

Changes in walking parameters of milking cows after hoof trimming

We examined the walking parameters of milking cows in relation to hoof trimming. The hooves of eight Holstein cows were trimmed. Two days before and after the hoof trimming, walking behavior, limb angles, back posture and vertical movement of the back while walking were measured. Walking rate (1.27 vs 0.87 m/s; P = 0.003), step length (1.46 vs 1.33 m; P = 0.009) and stepping rate (0.78 vs 0.65 steps/s; P = 0.046) were found to increase significantly after hoof trimming. Limb angles at the start and end of the support phase changed significantly after hoof trimming, thus indicating an improvement in the ‘on tiptoe’ appearance. The parameters relating to backline indicated that the marked arched shape of the back diminished after hoof trimming. Before hoof trimming, the positions of the withers varied widely and irregularly while walking; however, it changed after hoof trimming as the sinusoid curve and the range of vertical movement of withers decreased significantly. Overall, it has been quantitatively indicated that walking characteristics are improved after hoof trimming. The parameters used in the present study are expected to be useful for monitoring the walking characteristics of cows.     

Suspected spontaneous hyperadrenocorticism in a young experimental beagle dog

A 6-month-old female beagle dog, assigned to the low-dose group in a toxicity study, was evaluated for compound toxicity, and spontaneous hyperadrenocorticism was suspected. The animal had an externally apparent distended abdomen on clinical examination upon arrival. Pre-dose clinical pathology showed slightly higher erythroid parameters and stress leukogram on hematology; plasma biochemistry showed higher total protein, gamma-glutamyl transferase, total cholesterol, and triglyceride levels than the reference data. On necropsy, a prominent increase in adipose tissues of the subcutis and abdomen and increased weight of the adrenal gland and liver were observed. Histopathology revealed diffuse hyperplasia of adrenocortical cells in the zona fasciculata and reticularis, cortical atrophy of the thymus, and abundant glycogen accumulation in the hepatocytes. These findings were incidental and not test-substance-related. Electron microscopy of the adrenocortical cells in the zona fasciculata revealed decreased typical translucent lipid droplets, increased electron-dense lipid droplets, and abundant smooth endoplasmic reticulum and lysosomes. Additionally, increased numbers of various sizes and forms of mitochondria with tubular, vesicular, or lamellar cristae compared to that of normal animals were observed. These ultrastructural characteristics of the adrenocortical cells suggested hyperfunction. The pre-dose plasma cortisol levels were slightly higher than those of other females assigned to the toxicity study, while plasma adrenocorticotropic hormone levels were within the normal range. These findings indicate that hyperadrenocorticism is a possible cause of the systemic changes in this case.     

An intracytoplasmic injection of deionized bovine serum albumin immediately after somatic cell nuclear transfer enhances full-term development of cloned mouse embryos

In mouse somatic cell nuclear transfer (SCNT), polyvinylpyrrolidone (PVP) is typically included in the nuclear donor injection medium. However, the cytotoxicity of PVP, which is injected into the cytoplasm of oocytes, has recently become a cause of concern. In the present study, we determined whether bovine serum albumin deionized with an ion-exchange resin treatment (d-BSA) was applicable to the nuclear donor injection medium in SCNT as an alternative to PVP. The results obtained showed that d-BSA introduced into the cytoplasm of an enucleated oocyte together with a donor nucleus significantly enhanced the rate of in vitro development of cloned embryos to the blastocyst stage compared with that of a conventional nuclear injection with PVP in SCNT. We also defined the enhancing effects of d-BSA on the blastocyst formation rate when d-BSA was injected into the cytoplasm of oocytes reconstructed using the fusion method with a hemagglutinating virus of Japan envelope before oocyte activation. Furthermore, immunofluorescence experiments revealed that the injected d-BSA increased the acetylation levels of histone H3 lysine 9 and histone H4 lysine 12 in cloned pronuclear (PN) and 2-cell embryos. The injection of d-BSA before oocyte activation also increased the production of cloned mouse offspring. These results suggested that intracytoplasmic injection of d-BSA into SCNT oocytes before oocyte activation was beneficial for enhancing the in vitro and in vivo development of mouse cloned embryos through epigenetic modifications to nuclear reprogramming.     

Immunohistochemical analysis of the distribution of classical swine fever (CSF) viral antigen in boar-pig hybrids and pigs four weeks after infection

We detected the classical swine fever virus (CSFV) antigen in three boar-pig hybrids (hybrids) and three pigs. All animals were experimentally infected with CSFV strain JPN/27/2019 to optimize diagnostic sampling and risk assessment of virus dissemination. Two hybrids died 17- and 19-days post-inoculation (dpi). The other animals were euthanized at 28 dpi. The detection of CSFV antigen at 28 dpi in epithelial cells of the apocrine sweat and sebaceous glands in the skin, salivary glands, mucosal epithelial cells in the rectum, and epithelial cells in the kidney and urinary bladder, suggests that CSFV persists in these tissues and spreads via sweat, saliva, feces, and urine for at least 4 weeks. These findings reveal that hybrids and pigs represent a high risk of virus dissemination four weeks after infection with CSFV strain JPN/27/2019. Prominent CSFV antigens were also detected in hair follicles of the skin. These results suggest that postmortem sampling of animal skin may be effective for CSF diagnosis and can be used to develop a rapid and easy diagnostic method using hair follicles.