cFLIP regulates death receptor-mediated apoptosis in an ovarian granulosa cell line by inhibiting procaspase-8 cleavage

More than 99% of follicles in mammalian ovaries undergo atresia, but the mechanisms regulating the strict selection process are still unclear. Granulosa cell apoptosis is considered the trigger of follicular atresia, which occurs in advance of the death of an oocyte. Cellular FLICE-like inhibitory protein (cFLIP), a homologue of procaspase-8 (also called FLICE), is an intracellular anti-apoptotic protein. It is expressed in granulosa cells of porcine ovaries, where its levels decreases during follicular atresia. We hypothesized that cFLIP regulates granulosa cell apoptosis by acting as a pro-survival factor. In the present study, to further reveal the function of cFLIP in granulosa cells, we examined the anti-apoptotic mechanism of cFLIP using KGN, a human granulosa tumor cell line. Fas-mediated apoptosis was induced by co-treatment with anti-Fas antibody (CH-11), which acts as an agonist of Fas-ligand, and cycloheximide (CHX). When cFLIP was stably expressed in KGN cells following transfection of an expression vector, the Fas-mediated apoptosis was inhibited. Suppression of cFLIP by small interfering RNA (siRNA) spontaneously induced cell death. Silencing of cFLIP promoted cleavage of procaspase-8, and the cell death caused by cFLIP siRNA was completely blocked by a caspase-8 inhibitor (Z-IETD-FMK), indicating that cFLIP regulates apoptosis in KGN cells by inhibiting cleavage of procaspase-8. In conclusion, cFLIP is an essential pro-survival factor for granulosa cells, and it prevents granulosa cell apoptosis by inhibiting procaspase-8 activation.     

Effect of feeding wood kraft pulp on the growth performance,feed digestibility,blood components,and rumen fermentation in Japanese Black fattening steers

This study aimed to examine the effects of feeding kraft pulp (KP) on the growth performance, feed digestibility, and rumen fermentation of Japanese Black fattening steers. Ten Japanese Black fattening steers (aged 26 months) were randomly divided into control and KP groups. The control group (n = 5) was fed concentrate feed without KP, and the KP group (n = 5) was fed concentrate feed containing 10% KP. Both the groups were provided rice straw as roughage. The experiment was conducted over a period of 12 weeks. There was no significant difference in dry matter intake, daily body weight gain, and nutrient digestibility between both groups. No difference was observed in the ruminal concentrations of volatile fatty acids among the groups. At weeks 8 and 12 after the onset of the experiment, the acetate‐to‐propionate ratio in the ruminal fluid of the KP group was significantly higher than that of the control group. The average daily pH of ruminal fluid and activity of ruminal lipopolysaccharide did not differ between the groups. Our results suggested that the growth performance and feed digestibility in the Japanese Black fattening steers were not influenced by replacing concentrate feed with KP.     

Evaluation of three hormonal protocols for anovulatory lactating cows under regulations restricting the use of estrogenic compounds

When European Union regulations restricted the use of estrogenic compounds in food‐producing animals, refined hormonal protocols were no longer applicable for anovulatory cows. However, Ovsynch and its adaptations are routinely and uniformly applied to all cows regardless of ovarian function. To evaluate their efficacy on anovulatory cows, 143, 147 and 144 anovulatory cows received Ovsynch, Presynch and G6G protocols, respectively. In comparison, 150 cyclic cows were bred without using a synchronized protocol. Results showed that cows in the Presynch group had luteolysis responding to the last prostaglandin F_2α (PGF_2α) injection greater than the Ovsynch group. The serous progesterone levels at the first gonadotropin‐releasing hormone of Ovsych and the last PGF_2α injection was greater in the G6G group than the other two hormonal treatment groups. Concentrations of Ca²⁺ and total protein in cervical mucus in all three hormone‐treated groups before artificial insemination (AI) were significantly different from the controls. The G6G group obtained a greater pregnancy rate compared with Ovsynch and Presynch, but significantly less than the controls. For open cows in the Ovsynch group, estrus rate within 24 days after the first AI was significantly less than the controls. In conclusion, the G6G treatment resulted to better reproductive performance in anovulatory cows.     

Cover Image,Volume 16, Issue 3

The cover image, by K. Sakai et al., is based on the Original Article Association of tumour‐infiltrating regulatory T cells with adverse outcomes in dogs with malignant tumours, DOI: 10.1111/vco.12383 .

     

Spontaneous osteosarcoma in a rabbit ( Oryctolagus cuniculus)

A 6-year-old male cross-breed rabbit ( Oryctolagus cuniculus) was presented with lameness and severe swelling from the right shoulder to brachium. On 16-detector helical computed tomography images of the amputated right forelimb after being fixed in formalin, evident proliferative and destructive lesions of bone were observed. On histologic examination, the tumor was composed of proliferating neoplastic cells that resembled histiocytes, with abundant osteoid production. A large number of multinucleated giant cells were found throughout. This case was diagnosed as osteosarcoma by clinical, radiographic, and histologic findings. This is a rare case report of osteosarcoma in a rabbit consistent with canine predilection sites.     

Effect of emerged shipment on the physiological condition of the adductor muscle in adult giant lion's paw scallop Nodipecten subnodosus (Sowerby 1835)

Currently, there is an increasing interest in shipping live adult scallops to markets and broodstock to hatcheries; nevertheless, information about the shipping effect on live scallops physiology is very scarce. In a previous study, a method in emersion was developed to ship scallop seed out of water, but it was not known whether this method is useful to transport adult organisms. As a consequence, the purpose of this study was to evaluate the effects of transportation by the emersion method on the physiological status of the adductor muscle of adult giant lion's paw scallop. Live specimens were packaged in a container and transported in emersion for 11 h. Six scallops were frozen in a farm and a similar number were frozen as soon as they arrived to the laboratory. Physiological indices were determined in each lot and the survival was estimated 24 h after re‐immersion. As a result of the transportation, a significant loss (P<0.05) of total carbohydrates, glycogen, adenosine 5′‐triphosphate and adenilated energetic charge, and a significant increase (P<0.05) in the free amino acids concentration were observed. Eighty‐eight per cent survival was achieved; therefore, we conclude that this method is appropriate for shipping live adult scallops of Nodipecten subnodosus.     

Quantitative loop-mediated isothermal amplification method for the detection of <Emphasis Type="Italic">Vibrio nigripulchritudo</Emphasis> in shrimp

Vibrio nigripulchritudo is considered one of the major pathogens threatening shrimp aquaculture. In this study, we developed a novel and highly specific quantitative loop-mediated isothermal amplification (Q-LAMP) assay. A set of four specific primers were designed targeting the V. nigripulchritudo intergenic spacer region. The reaction time and temperature were optimized for 60 min at 63°C. Quantitative analysis was then performed by measuring the turbidity of the reaction solution using a real-time turbidimeter, allowing for quantification of the initial DNA concentration with a sensitivity of 10² copy numbers equivalent to 2.3 colony forming units/ml or 0.3 fg/μl. The LAMP assay was able to specifically detect two representative strains of V. nigripulchritudo, whereas other Vibrio and non-Vibrio species were not amplified. A standard curve was generated for V. nigripulchritudo by plotting the threshold time (T _t) versus the log of bacterial number. A high correlation coefficient (R ² = 0.9749) was observed for the Q-LAMP reaction. In conclusion, Q-LAMP assay is a sensitive, rapid, and simple tool that can be used for the detection and quantification of V. nigripulchritudo in shrimp, thereby facilitating surveillance of vibriosis infection.     

Monitoring cotton (<Emphasis Type="Italic">Gossypium hirsutum</Emphasis> L.) germination using ultrahigh-resolution UAS images

Examination of seed germination rate is of great importance for growers early in the season to determine the necessity for replanting their fields. The objective of this study was to explore the potential of using unmanned aircraft system (UAS)-based visible-band images to monitor and quantify the cotton germination process. A light-weight UAS platform was used, which carried a consumer-grade red, green, and blue camera stabilized by a built-in gimbal system. In order to obtain ultrahigh image resolution during the germination stage, the UAS platform was flown at an altitude of approximately 15–20 m above ground. By applying the structure-from-motion (SfM) algorithm, the images were rectified and orthographically mosaicked with a ground sampling distance of approximately 6–9 mm/pixel. A novel solution was then developed for calculating the average plant size and the number of germinated cotton plants according to the leaf polygons extracted from the orthomosaic images. By using the estimated number of germinated cotton plants, the plant density and the cumulative germination rate can also be estimated in a straightforward manner using field-specific parameters. An assessment of the proposed solution was conducted by comparing the estimated number of the germinated cotton plants against ground observation data collected from six cotton row segments. The results demonstrated that the average estimation accuracy achieved 88.6% in terms of identifying the number of the germinated cotton plants. The accuracy may be further improved if images with near infrared band are employed.