Contrasting Size Distributions among the Wing Spots of a Pierid Butterfly Suggest Different Selective Histories.

Males of the small cabbage butterfly Pieris rapae crucivora have two dark or melanic spots in the central white area of each dorsal forewing, an anterior spot (aS) and a posterior spot (pS). We used characteristics of the size distributions of these spots to infer how selection has acted on them during their evolution. Our study reveals that the aS size distribution is normal while that of pS is very right-skewed. Moreover, aS size is larger and less variable than pS size. These results suggest that the aS has been under stabilizing selection while the pS has not. The context in which this selection is acting is not yet clear, nor is it clear why pS persists as a wing marking.

Effect of vitamin K3 supplementation on immunoglobulin G concentration in colostrum of periparturient Holstein dairy cows.

This study was to examine the effects of dietary vitamin K (VK) 3 supplementation on immune-related substances in milk, oxidative stress indices in plasma and VK1, and menaquinone 4 (MK-4) in plasma and milk in periparturient dairy cows. Forty healthy perinatal Holstein-Friesian dairy cows were used in this study. Twenty-one animals were randomly selected and categorized into the VK3 supplemented (50 mg/day/head as VK3) group; the remaining 19 were categorized into the control group. On day 3 after calving, blood and milk were sampled, and their chemical components were determined. The VK3 supplemented group had significantly higher menaquinone 4 levels in plasma and milk on day 3 postpartum than the control group. In addition, there was a significant increase in the immunoglobulin G (IgG) level in milk. VK3 may be absorbed from the gastrointestinal tract and converted to MK-4, the biologically active form of VK, in the mammary gland and other tissues. It was thought that the increase in MK-4 level in plasma and milk induced an increase in the concentration of IgG in milk. VK3 supplementation to periparturient dairy cows may contribute to the production of colostrum with high concentrations of IgG and MK-4.

Effects of dietary vitamin K3 supplementation on vitamin K1 and K2 (menaquinone) dynamics in dairy cows.

The effect of dietary vitamin K3 (VK3) on ruminant animals is not fully investigated. The aim of this study was to examine the effects of dietary VK3 on lactation performance, rumen characteristics, and VK1 and menaquinone (MK, or VK2) dynamics in the rumen, plasma, and milk of dairy cows. Eight Holstein dairy cows in late lactation periods were used in two crossover trials including a control (nontreatment) and a 50 or 200 mg/day (d) VK3 supplementation group. After 14 days, plasma, ruminal fluid, and milk were sampled and their VK1 and MKs contents were measured using fluorescence-high-performance liquid chromatography (HPLC). Milk production was unchanged after feeding 50 mg/day VK3 but marginally decreased after feeding 200 mg/day VK3. The molar ratio of propionate in ruminal fluid was significantly increased on feeding 200 mg/day VK3. Additionally, MK-4 concentrations significantly increased in both plasma and milk after VK3 feeding (50 and 200 mg/day). In ruminal fluid, MK-4 concentrations increased after 200 mg/day VK3 feeding. These results suggest that VK3 may be a good source of MK-4, the biologically active form of VK, in Holstein dairy cows during their late lactation periods. This study provides a basis for understanding the physiological role of VK in dairy cows.

Effects of ruminal administration of soy sauce oil on functional fatty acids in the rumen, blood and milk of dairy cows.

OBJECTIVE: Soy sauce oil, a byproduct of whole soybean processing by the soy sauce industry, was evaluated as a source of linoleic acid for dairy cows for the purpose of manipulating the composition of milk. METHODS: Eight dairy Holstein cows fitted with rumen cannulas were used for ruminal administration of soy sauce oil for a 28-day period using a 4×4 Latin square study design with 4 doses (0, 200, 400, and 600 g soy sauce oil/d). RESULTS: Although dry matter intake and milk yield were not affected by soy sauce oil administration, ruminal concentrations of total volatile fatty acids and acetate were decreased, specifically at 600 g/d administration. While milk fat percentage was decreased with administration of soy sauce oil, proportions of linoleic, vaccenic and conjugated linoleic acids in the rumen, blood and milk were increased with increasing soy sauce oil dose. CONCLUSION: These results suggest that soy sauce oil feeding could be useful for improving milk functionality without adverse effects on animal production performance when fed at less than 400 g/d.

Cluster analysis to evaluate disease risk in periparturient dairy cattle.

Predicting periparturient disease risk is of immense value to the dairy industry. Periparturient diseases are interrelated with each other; however, predicting the onset risk of these diseases has predominantly been based on a single blood parameter for a single disease. This study examined a new diagnostic method to predict the risk of periparturient diseases. We conducted cluster analysis of multiple blood constituents from 20 Holstein cattle at 1 week post-partum, and the cattle were divided into two groups, A or B. We then compared the periparturient and early-lactation blood constituents of these groups. Group B had significantly higher 3-hydroxybutyric acid concentrations and were suspected to have subclinical ketosis. Group B also had significantly lower calcium concentrations, with a tendency for subclinical hypocalcemia. We also performed discriminant analysis using blood parameters at 1 week post-partum, which grouped the population into the same two groups as the cluster analysis based on three variables: inorganic phosphorus, calcium, and either phospholipids or total cholesterol. We further showed that these discriminant functions could be used to predict the risk of periparturient disease even before parturition. Our results indicate that cluster analysis with multiple blood constituents is useful for predicting periparturient disease risks.

Effect of ruminal administration of soy sauce oil on rumen fermentation, milk production and blood parameters in dairy cows.

OBJECTIVE: To evaluate soy sauce oil (a by-product of making whole soybean soy sauce) as a new dietary lipid source, a large amount of soy sauce oil was administered into the rumen of dairy cows. METHODS: Four Holstein dairy cows fitted with rumen cannulae were used in a 56-day experiment. Ruminal administration of soy sauce oil (1 kg/d) was carried out for 42 days from day 8 to day 49 to monitor nutritional, physiological and production responses. RESULTS: Dry matter intake and milk yield were not affected by soy sauce oil administration, whereas 4% fat-corrected milk yield and the percentage of milk fat decreased. Although ruminal concentration of total volatile fatty acids (VFA) and the proportion of individual VFA were partially affected by administration of soy sauce oil, values were within normal ranges, showing no apparent inhibition in rumen fermentation. Administration of soy sauce oil decreased the proportions of milk fatty acids with a carbon chain length of less than 18, and increased the proportions of stearic, oleic, vaccenic and conjugated linoleic acids. Conjugated linoleic acid content in milk became 5.9 to 8.8 times higher with soy sauce oil administration. Blood serum concentrations of non-esterified fatty acid, 3-hydroxybutyric acid, total cholesterol, free cholesterol, esterified cholesterol, triglyceride and phospholipid increased with administration of soy sauce oil, suggesting a higher energy status of the experimental cows. CONCLUSION: The results suggest that soy sauce oil could be a useful supplement to potentially improve milk functionality without adverse effects on ruminal fermentation and animal health. More detailed analysis is necessary to optimize the supplementation level of this new lipid source in feeding trials.

Time course of changes in antioxidant activity of milk from dairy cows fed a trehalose-supplemented diet.

This study was to investigate the time course of changes to the antioxidant activity of milk from cows fed a trehalose-supplemented diet, and to determine possible underlying mechanisms for observed changes. Six Holstein cows were used, and subjected to two experimental feeding periods consisting of a 1% trehalose-supplemented diet for 10 days, followed by a basal diet only (no trehalose) for 10 days. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) radical scavenging activities in milk were gradually increased during the trehalose supplementation period and were highest at the end of the second period. However, trehalose was not detected in the milk and plasma of dairy cows fed a diet supplemented with trehalose for 10 days, indicating that the increased antioxidant activity in the milk of trehalose-fed cows is not due to the direct transfer of trehalose to the milk. Plasma DPPH activities exhibited a similar time course to that seen for milk. Relative superoxide dismutase (SOD) activities in the rumen were higher 3 days after the end of trehalose supplementation than at any other time during the experimental periods. These results suggested that the improved antioxidant activity in milk and plasma of cows fed a trehalose-supplemented diet was due to improved ruminal relative SOD activity.

Changes in circulating adiponectin and metabolic hormone concentrations during periparturient and lactation periods in Holstein dairy cows.

Although our previous report demonstrated that adiponectin and AdipoR1 gene expressions changed among different lactation stages in the bovine mammary gland, its in vivo kinetics remain unclear in ruminant animals. In this study, we investigated the changes in circulating concentrations of adiponectin, as well as other metabolic hormones and metabolites, (i) during the periparturient period and (ii) among different lactation stages, in Holstein dairy cows. In experiment 1, serum adiponectin concentrations increased after parturition. Serum insulin concentrations were lower in the postpartum than prepartum period, whereas serum growth hormone (GH) concentrations increased in the postpartum period. Serum nonesterified fatty acids (NEFA) levels were increased during the postpartum period and were dependent on the parity. In experiment 2, there was no significant difference in plasma adiponectin concentrations among lactational stages. Plasma insulin concentrations tended to be lower in early lactation while plasma GH levels tended to be higher. Plasma NEFA concentrations were significantly lower in mid- and late-lactation stages than non-lactation stages. These findings indicate that elevation of serum adiponectin might be involved in energy metabolism just around parturition, and might exert its action through regulation of receptor expression levels in target tissues in each lactational stage in Holstein dairy cows.

Effects at early stage of life of elevated milk replacer feeding on growth rate, plasma IGF-I concentration and intestinal nutrient transporter expression in Holstein bull calves.

In order to evaluate the effects of an elevated amount of modified milk replacer on body weight, daily gain, starter intake, plasma endocrine parameters and expression of nutrient transporters in small intestinal epithelia, Holstein bull calves (n=24) were fed for 60days either with the usual amount of 24% crude protein (CP) and 20% fat milk (CF) replacer (C group), or with a double amount of a modified milk replacer of 28% CP and 16% CF (E group). Body weight from D20 to D60 and daily gain before D40 was greater or tended to be greater for the E group than the C group. Plasma concentrations of insulin-like growth factor-1 (IGF-I) and insulin were greater for the E group than the C group on D28 but not on D56, without changing plasma growth hormone levels. Gene expression for sodium-dependent glucose transporter 1 and fatty acid translocase (CD36) was altered in day- and intestine-dependent manners. From these findings, we conclude that an elevated intake of milk replacer given up to 40days old is sufficient to enhance body weight, which may be associated with increased plasma IGF-I concentrations, in Holstein bulls.

Serotonin-induced mate rejection in the female cabbage butterfly, Pieris rapae crucivora.

Virgin female cabbage butterflies, Pieris rapae crucivora, accept and mate with courting males, whereas mated females reject them and assume the "mate refusal posture". This study tested whether the biogenic amines, serotonin (5HT), dopamine (DA), and octopamine (OA), were responsible for this change in behavior. The results showed that 2-3-day-old virgin females fed with 5HT rejected courting males significantly more frequently compared with controls fed on sucrose. In contrast, the proportions of courting males rejected by virgin females fed with either DA or OA did not differ from sucrose-fed controls. Oral application of each amine resulted in significantly increased levels of the amine applied (or its metabolite) in the brain. The results strongly suggest that 5HT or a 5HT metabolite may be responsible for the post-mating change in behavioral response of 2-3-day-old virgin females to courting males. Similar effects of 5HT treatment were observed in 6-8-day-old virgin females, but in this case the results were only marginally different from the controls, suggesting that the effect may decline with increasing female age.

Effect of supplementation of soy sauce oil and Ca salts of fatty acids on rumen fermentation, milk production and conjugated linoleic acid in milk of dairy cows.

Eight cows were used to evaluate the effects of supplementation of soy sauce oil (SO) or Ca salts of fatty acids (FA) on rumen fermentation and milk production. The control diet (CO) consisted mainly of hay, corn silage and a concentrate. In the experimental diets, 400 g/day per cow of SO or FA (soybean oil and rapeseed oil) was supplemented to the CO diet. Experimental period for the three treatments was 14 days, and milk samples were taken during the last 2 days and rumen sample was taken on the last day. Dry matter intake was not affected by the treatments. The number of rumen protozoa at 0 h increased by SO and FA diets. Total volatile fatty acids at 2 h after feeding of SO diet was decreased compared to CO. The milk composition yield did not differ among treatments, although the percentages of fat and protein were decreased by SO and FA diets. The proportions of C8-C16 fatty acids in milk fat decreased, and those of C18 increased by SO and FA diets. The proportion of cis-9, trans-11 conjugated linoleic acid in milk fat by SO and FA diets increased by 120% and 135%, respectively. In spite of the slight suppression of rumen fermentation by SO diet, negative effects on feed intake and milk production were not detected.

Saturated fatty acids stimulate and insulin suppresses BMCP1 expression in bovine mammary epithelial cells.

Brain-specific uncoupling proteins such as uncoupling protein 4 (UCP4) and brain mitochondrial carrier protein 1 (BMCP1; also known as UCP5) were identified by computational analysis for expressed sequence tag and hybridization screening. Both were detected at the mRNA level by RT-PCR in cloned bovine mammary epithelial cells (bMEC) and lactating bovine mammary glands. Physiological concentrations of saturated fatty acids (stearate and palmitate), but not unsaturated fatty acids (oleate and linoleate), induced up-regulation of BMCP1 mRNA in bMEC. Treatment with insulin induced down-regulation of UCP4 and BMCP1. These results suggest that UCP4 and BMCP1 are regulated by insulin and/or fatty acids in mammary epithelial cells and lactating mammary glands, and thereby may play an important role in lipid and energy metabolism.

Inaccurate mate recognition as a mating strategy of a 'pioneer male'.

Heterosubspecific mating experiments were carried out between two subspecies of cabbage butterflies, British Pieris rapae rapae and Japanese P. rapae crucivora, to examine how accurately males recognize the mates. The two subspecies are different in that the wings of female P. rapae rapae reflect little UV light, whereas those of female P. rapae crucivora are strongly UV-reflective. The wing colouration of P. rapae crucivora involving UV is believed to be critical in mate recognition. The results showed that males of both subspecies displayed mating behaviours, to and copulated with, females of both subspecies. Furthermore, P. rapae crucivora males exhibited mating behaviours and attempted to copulate with females of Pieris melete with low UV reflectance which are critically different from P. rapae crucivora females with high UV reflectance. Based on these findings, we propose the "pioneer male" hypothesis, which argues that such inaccurate mate recognition may sometimes be selectively beneficial for males and thus an adaptive mating strategy. The "pioneer male" was discussed in terms of its possible role in the evolution.

Saturated fatty acids stimulate and insulin suppresses CIDE-A expression in bovine mammary epithelial cells.

Cell death-inducing DNA fragmentation factor-alpha-like effector A (CIDE-A) was first identified by its sequence homology with the N-terminal domain of DNA fragmentation factor (DFF). CIDE-A negatively regulates the activity of uncoupling protein 1 (UCP1) in brown adipose tissue. CIDE-A and UCP1 mRNA were detected by RT-PCR in cloned bovine mammary epithelial cells (bMEC) and lactating bovine mammary glands. Physiological concentrations of saturated fatty acids (stearate and palmitate), but not unsaturated fatty acids (oleate and linoleate) induced up-regulation of CIDE-A mRNA in bMEC. Treatment with insulin (5-10 ng/ml) induced down-regulation of CIDE-A and UCP1. The expression levels of CIDE-A and UCP1 mRNA in bovine mammary glands at various stages of the lactation cycle were determined by quantitative RT-PCR analysis. CIDE-A mRNA expression at peak lactation (2 months after parturition) was significantly higher than at dry off and non-pregnancy but not late lactation. These results suggest that CIDE-A and UCP1 are regulated by insulin and/or fatty acids in mammary epithelial cells and lactating mammary glands, and thereby play an important role in lipid and energy metabolism.

Short-chain fatty acid signaling pathways in bovine mammary epithelial cells.

GPR41 and 43 have recently been identified as G-protein-coupled cell-surface receptors for short-chain fatty acids (SCFAs). Bovine orthologs of GPR41 and 43 (bGPR41 and 43) mRNA were detected by RT-PCR in cloned bovine mammary epithelial cells (bMEC) and various lactation stages of bovine mammary gland. Acetate and propionate caused an increase in intracellular Ca(2+) concentrations in these cells that was blocked by the treatment with pertussis toxin (PTX). SCFAs significantly reduced forskolin-induced cAMP concentrations in these cells. The phosphorylation of mitogen-activated protein kinase (MAPK) p38 was selectively increased by SCFAs. The downstream substrate heat shock protein 27 (HSP27) was also phosphorylated by SCFAs at Ser-78 and -82, but not -15. These results suggest that bGPR41 mainly, but not bGPR43, mediate SCFA signaling in mammary epithelial cells and thereby plays some important role in mammary gland.

Effects of GH gene polymorphism and sex on carcass traits and fatty acid compositions in Japanese Black cattle.

To investigate the effects of bovine growth hormone (bGH) gene polymorphism on carcass traits and fatty acid compositions in Japanese Black cattle caused by nucleotide substitution of CTG (allele A)/GTG (allele B) at codon 127 and of ACG (allele A and B)/ATG (allele C) at codon 172 of bGH, GH genotypes of 135 cattle were determined using allele specific-multiplex polymerase chain reaction (PCR). Allele A gave greater rib thickness and lower melting point of fat (MP) while allele B gave higher C18:1% (P < 0.05). Allele C gave higher C18:1, monounsaturated fatty acid (MUFA), unsaturated fatty acid (USFA) percentages (P < 0.05). It also gave lower saturated fatty acid (SFA) percentages, higher MUFA/SFA and USFA/SFA ratios, and lower MP (P < 0.05). Interactions of sex and GH alleles were analyzed. In heifers, allele A gave higher carcass weight, daily carcass gain, rib eye area, rib thickness, subcutaneous fat thickness, and BMS while allele B gave greater rib eye area and rib thickness (P < 0.05). Allele C gave higher C18:1 (P < 0.01), MUFA (P < 0.01), USFA percentages (P < 0.05) and MUFA/SFA and USFA/SFA ratios (P < 0.01), and lower C16:0 and SFA percentages (P < 0.05) and MP (P < 0.01). GH gene polymorphism affected carcass traits and fatty acid compositions although the effects were more pronounced in heifers.

Effects of 5'-uridylic acid feeding on postprandial plasma concentrations of metabolites and metabolic hormones in pre-weaning goats.

5'-Uridylic acid (UMP), which is present at high concentrations in cow's colostrum, has been shown to cause a reduction in increased plasma levels of insulin and glucose after ingestion of milk replacer in pre-weaning calves. However, the precise mechanisms of UMP action have not been investigated, and its action has not been investigated in other pre-weaning ruminants. In order to demonstrate whether UMP causes changes in postprandial metabolic and hormonal parameters in pre-weaning goats, 11 Saanen kids were given milk replacer (twice a day) without (n = 5) or with (n = 6) UMP (1 g for each meal, 2 g/day for each head) for 14 days. Analysis of blood samples taken in the morning of day 14 demonstrated that the feeding of milk replacer with UMP abolished the significant changes in postprandial plasma glucose, NEFA, GH and insulin concentrations induced by feeding of milk replacer alone, and demonstrated a tendency to increase IGF-I levels. However, there was no significant difference between the two groups at any sampling time. We conclude that UMP feeding with milk replacer showed a tendency to blunt the postprandial changes in levels of some plasma metabolites and hormones that are induced by replacer alone in pre-weaning goats.

Better mate in the shade: enhancement of male mating behaviour in the cabbage butterfly, Pieris rapae crucivora, in a UV-rich environment.

Ultraviolet (UV) vision is widespread in a variety of animals, playing important roles in behaviours such as foraging and reproduction. Despite accumulated information about UV vision and UV-dependent behaviours of animals, little is known about the effect of temporal changes and local variations in UV light on UV-dependent behaviour. Here we report the mating behaviour of male cabbage butterfly, Pieris rapae crucivora, in environments with varying content of UV light. We first confirmed that the relative UV content is higher in shaded places than in sunny places. We furthermore arranged experimental areas with varying UV contents in the field, where we compared three aspects of male mating behaviour: visual localization of females, female-searching flight and copulation success rate. In all aspects males performed more actively in UV-rich environments: males searched females for longer, approached females preferentially in the shade and copulated there more frequently. Apparently, female-searching males detect females more easily in a UV-rich environment. The present findings should be taken into consideration when UV-dependent behaviours, visual mate choice in particular, are studied.

Regulation of uncoupling protein 2 expression by long-chain fatty acids and hormones in bovine mammary epithelial cells.

Although mammary epithelial cells are known to synthesize and accumulate triacylglycerol (TAG) in order to produce milk lipid in the cytosol, lipid and energy metabolism is still not fully understood. In this study, we assessed the effects of long-chain fatty acid (LCFA) on the accumulation of cytosolic TAG and uncoupling protein (UCP) 2 in cloned bovine mammary epithelial cells (bMEC). LCFAs significantly raised the expression of UCP2 mRNA and the accumulation of TAG. We observed the rapid elevation in UCP2 shown at 6h after LCFA treatment. Insulin (5-50 ng/ml) or dexamethasone (500 nM) significantly suppressed the expression of UCP2 mRNA. These results suggest that UCP2 play an important role of lipid and energy metabolism in mammary epithelial cells.

Regulation of hormone-sensitive lipase expression by saturated fatty acids and hormones in bovine mammary epithelial cells.

Hormone-sensitive lipase was firstly identified as an epinephrine-induced lipase in adipocyte. HSL mRNA was detected by RT-PCR in cloned bovine mammary epithelial cells (bMEC) and bovine lactating mammary gland. Saturated fatty acids (stearate and palmitate), but not unsaturated fatty acids (oleate and linoleate) induced up-regulation of HSL mRNA in a time- and concentration-dependent manner in bMEC. Treatment with insulin (5-10 ng/ml), dexamethasone (50-250 nM) or GH (50 ng/ml) induced down-regulation of HSL. These results suggest that HSL was regulated by fatty acids and some hormones in mammary epithelial cells and thereby play an important role of lipid and energy metabolism.