Functional insulin‐like factor 3 (INSL3) hormone‐receptor system in the testes and spermatozoa of domestic ruminants and its potential as a predictor of sire fertility

Insulin‐like factor 3 (INSL3) is essential for fetal testis descent, and has been implicated in the testicular and sperm functions in adult males; however, similar functions in domestic ruminants remain largely unknown. This study investigated the functional INSL3 hormone‐receptor system in adult ruminant testes and spermatozoa, and explored its potential to diagnose the fertility of sires. Testes and spermatozoa were obtained from fertile bulls, rams and he‐goats, whereas subfertile testes and spermatozoa were obtained only from bulls. As expected, INSL3 was visualized in Leydig cells, while we clearly demonstrated that the functional receptor, relaxin family peptide receptor 2 (RXFP2), enabling INSL3 to bind was identified in testicular germ cells and in the sperm equatorial segment of bulls, rams and he‐goats. In comparison to fertile bulls, the percentage of INSL3‐ and RXFP2‐expressing cells and their expression levels per cell were significantly reduced in the testes of subfertile bulls. In addition, the population of INSL3‐binding spermatozoa was also significantly reduced in the semen of subfertile bulls. These results provide evidence for a functional INSL3 hormone‐receptor system operating in ruminant testes and spermatozoa, and its potential to predict subfertility in sires.     

The effect of pelleting on in situ rumen degradability of compound feed containing brown rice for dairy cows

This study was conducted to determine the effect of pelleting on in situ dry matter degradability of pelleted compound feed containing brown rice for dairy cows. Mash feed of the same composition was used as a control and the in situ study was conducted using three non‐lactating Holstein steers fitted with a rumen cannula. The feeds contained 32.3% brown rice, 19.4% rapeseed meal, 11.4% wheat bran and 10.6% soybean meal (fresh weight basis). Except for moisture content, the chemical composition of the feed was not affected by pelleting. In situ dry matter disappearance of the feed increased from 0 to 2 h and after 72 h of incubation with pellet processing. Integration of the dry matter disappearance values over time revealed that degradability parameter a (soluble fraction) increased with pellet processing, whereas parameter b (potentially degradable fraction) decreased. Parameter c (fractional rate of degradation) and effective degradability (5% passage rate) were not affected by pellet processing. We concluded that pellet processing promotes rumen degradability at early incubation hours when the pelleted feed contains brown rice.     

Symptoms and molecular characterization of apple dimple fruit viroid isolates from apples in Japan

Apple dimple fruit viroid was detected from an apple tree (‘Jonagold’) bearing apples with mild dapple apple symptom. The isolates in Japan were distinct from those in apples in Italy and China and in fig in Italy. Graft-inoculation experiments showed that the symptoms were variable depending on the cultivar, and the symptom on ‘Starking Delicious’ was virtually similar to those reported in Italy. Symptoms induced by apple dimple fruit viroid were similar in part to those by apple fruit crinkle viroid or apple scar skin viroid, indicating that they cannot be discriminated by symptoms on any specific variety.     

Ectomycorrhizal fungal community of naturally regenerated Pinus thunbergii seedlings in a coastal pine forest

To add to knowledge of ectomycorrhizal (ECM) community structure on the roots of Pinus thunbergii seedlings in a Japanese coastal forest, we sampled naturally regenerated current-year and 1- to 5-year-old seedlings. We classified the 667 root tips on current-year seedlings and the 1,927 root tips on older seedlings into 13 phylotypes based on morphological and genetic analyses. Cenococcum geophilum, members of the families Clavulinaceae, Russulaceae, or Thelephoraceae or the genus Trichophaea, were indicated to be fungi forming P. thunbergii ectomycorrhizas. Among them, C. geophilum and Clavulinaceae sp. 1 were the most or second most dominant species. A species accumulation curve based on the number of samples nearly reached a plateau, with observed species richness equal to 11 species and the Jackknife2 and Chao2 richness estimators indicating 14 and 12 species, respectively. In addition, Simpson’s 1/D was 3.89 and Shannon–Wiener’s H′ was 1.71, indicating a relatively low taxonomic diversity. There was no significant difference in the ectomycorrhizal formation rate, or the occurrence frequency and the number of ECM phylotypes between current-year and older seedlings. These results indicated that less diverse fungi were involved in ectomycorrhizal formation on coastal pine seedlings compared with those in comparable inland forests in the study area.     

Pathogenicity of Plasmodiophora brassicae populations from small, spheroid, resistant-type clubroot galls on roots of clubroot-resistant cultivars of Chinese cabbage (Brassica rapa L. subsp. pekinensis)

A resistant type of small, spheroid clubroot galls (SSGs) containing resting spores formed on the root surface of clubroot-resistant (CR) cultivars of Chinese cabbage (Brassica rapa L. subsp. pekinensis) inoculated with an avirulent population of Plasmodiophora  brassicae. Populations isolated from the SSGs severely affected a common (susceptible) cultivar but did not have the typical pathogenicity on CR cultivars, indicating similarity in pathogenicity between SSG and original spore populaions. Populations pathogenic on CR cultivars were not detected from SSGs. Therefore, the ability of the avirulent population among the SSGs to form resting spores may need to be considered to achieve clubroot control in common cultivars of crucifers.     

Muscle fiber types,growth and development in the whole myotome of cultured Pacific bluefin tuna Thunnus orientalis

Following the successful development of Pacific bluefin tuna (PBT) aquaculture, it is of considerable importance to determine the muscle fiber types and their growth patterns for future development. Muscle fiber profiles of dorsal ordinary, lateral ordinary (LO) and dark muscles and their growth patterns in PBT from 3.0 to 54.3 kg body weight were studied. Muscle fibers were histochemically stained for NADH-diaphorase and myosin adenosine triphosphatase activity (mATPase), and immunohistochemically stained with S-58 slow-muscle myosin antibody. All muscle fibers in dorsal and LO muscles showed low NADH-diaphorase activity, and acid-labile (pH 4.0 or 4.3) and alkali-stable mATPase activity. In LO muscle adjacent to dark muscle, three intensities of mATPase activity were observed after acid pre-incubation at pH 4.5 or 5.0, and the activity was related to the muscle fiber diameter. In dark muscle, all small and some large fibers stained intensely for NADH-diaphorase activity, related to their high aerobic metabolism. The high-active fibers with NADH-diaphorase in dark muscle were positive for S-58 antibody. Some large fibers in dark muscle showed intermediate NADH-diaphorase activity and high mATPase activity after alkali pre-incubations. These are fast-twitch oxido-glycolytic fibers in dark muscle and transformed to red muscle fibers with increasing body weight.     

Brewing Performance of Malted Lipoxygenase‐1 Null Barley and Effect on the Flavor Stability of Beer

We examined the malting and brewing performances of a lipoxygenase‐1 (LOX‐1) null line of barley (Hordeum vulgare L.). The LOX‐normal malt and the LOX‐null malt were prepared from F₄ populations derived from a single cross. We could not observe any major differences in the general malt characteristics between the two malts. A brewing trial was performed using these malts. The analysis of the wort and beer revealed that the absence of LOX‐1 had little effect on the general characteristics of the wort and beer. In contrast, beer made from the LOX‐null malt showed reduced levels of beer‐deteriorating substances, trans‐2‐nonenal (T2N), and trihydroxyoctadecenoic acid (THOD). In the sensory evaluation, well‐trained panel members recognized the significant superiority of the aged LOX‐null beer in terms of staleness. These results show that the LOX‐1 null barley line can be effectively used to improve the flavor stability of beer without changing the other important beer qualities.     

Female sex steroid residues in the tissues of steers treated with progesterone and oestradiol-17 beta.

Progesterone, oestrone and oestradiol-17 beta levels in the muscle, fat, liver, kidney and plasma of steers treated with Synovex S, untreated steers, and cows during the normal oestrous cycle, were examined. Progesterone levels in female tissues reached a maximum in the luteal phase and fell to a minimum in the follicular phase. Oestrogen levels (oestrone and oestradiol-17 beta) did not change during the cycle. Progesterone and oestrogen levels in the tissues of steers treated with Synovex S and untreated steers were not statistically different. Tissue progesterone levels in steers were much lower than those in cows during the luteal phase. The highest oestrogen levels were found in the fat of female animals. These results indicate that residue levels of progesterone and oestrogen in the muscle and fat of steers treated with Synovex S were within the physiological range, and lower than those of cows.     

Effects of environmental temperature on heat production associated with food intake and on abdominal temperature in laying hens.

An experiment was carried out to verify the relationship between the heat production associated with food intake and environmental temperature. Four laying hens were trained to eat two meals per day and were kept under artificial illumination with dark, dim and light periods. 2. Metabolic heat production was measured using two open-circuit respiratory chambers. Abdominal temperatures were measured, using thermocouples, at environmental temperatures of 12, 16, 20, 24, 28, 32 and 36 degrees C and at food intakes of 90, 60, 30 and 0 g/d. 3. The rate of heat production was dependent on both environmental temperature and food intake. Increasing environmental temperature resulted in a decrease in total metabolic rate at any food intake, indicating that heat production associated with food intake was not directly linked with thermoregulation at low environmental temperatures. 4. Abdominal temperature varied little with either food intake or environmental temperature below 28 degrees C. Above 28 degrees C, abdominal temperature increased with both environmental temperature and quantity of food, indicating that the heat production associated with food intake adds to the heat load at high environmental temperatures. 5. Both heat production and abdominal temperature declined with decreasing light intensity and increased before feeding time. These effects were considered to result from changes in physical activity.     

Elevated levels of tumor necrosis factor-α, (TNF-α) and interleukin-6 (IL-6) activities in the sera and milk of cows with naturally occurring coliform mastitis

The presence of tumor necrosis factor-α (tnf-α) and interleukin-6 (il-6) activities was determined in milk and serum of cows with naturally occurring coliform mastitis (cfm). tnf-α was detected in the sera from 26 of 32 cows with cfm. tnf-α levels were higher in the sera than in the milk. il-6 was high in the sera of surviving cfm animals, but was low in animals that died and in healthy controls. Furthermore, the mean level of il-6 was 20-fold higher in the milk than in the sera of mastitic cows. The level of il-6 in the serum was correlated to that in the milk in individual animals. The presence of il-6 and tnf-α in the sera appears to relate to severe clinical condition of cfm, in the milk whereas they may play a role in generating inflammation of the mammary gland.