A direct time-resolved fluoroimmunoassay (TR-FIA) for measuring plasma estradiol-17beta concentrations in cattle

A direct Time-Resolved Fluoroimmunoassay (TR-FIA) system for measuring estradiol-17beta (E(2)) in bovine plasma was developed and evaluated. A 100 microl sample of bovine plasma was used for a TR-FIA without prior extraction and purification. The dose-response curves of reference standards ranged from 0.0625 to 10 pg/well. The minimum detectable concentration of this assay system was 0.625 pg/ml, and 19 pg/ml of E(2) caused a 50% reduction of maximum binding. The intra- and inter-assay coefficients of variation were 10.2 and 17.4%, respectively. The plasma E(2) concentrations measured by direct TR-FIA correlated closely with those measured after extraction (r=0.939). The results in the present study indicate that the TR-FIA reagent for E(2), designed for human research can also be utilized, with some modification, for direct assaying in bovine plasma. This assay type seems to fulfill the requirements for safety, sensitivity, specificity, reproducibility and practical convenience.     

Optimal selection method for establishing an inbred strain of laboratory animals with high performance for litter size at weaning

The optimal selection method was investigated for establishing an inbred strain of laboratory animals with high performance for litter size at weaning (LSW). A Monte Carlo computer simulation was used to assess the effects of our selection methods on the genetic change of LSW under the continuous use of full‐sib mating for 20 generations. Smaller number of growing animals of each sex per litter and genetic evaluation for selection using a BLUP animal model increased LSW. Use of information on another trait genetically related to LSW, larger population size, and greater number of generations for random selection before starting full‐sib mating were useful for establishing an inbred strain of laboratory animals with high performance for LSW. It was concluded that LSW can be increased by directional selection when establishing inbred strains.     

Characterization and expression of non‐polymorphic liver expressed antimicrobial peptide 2: LEAP‐2 in the Japanese quail,Coturnix japonica

Liver‐expressed antimicrobial peptide 2 (LEAP‐2) is a cationic peptide that plays an important role in innate immunity for host defense. The aim of this study was to characterize the LEAP‐2 gene in the Japanese quail (Coturnix japonica). Japanese quail LEAP‐2 (CjLEAP‐2) was identified from the Japanese quail draft genome database by a local BLAST analysis using chicken LEAP‐2 (GgLEAP‐2). The exon‐intron structure of CjLEAP‐2, analyzed from three quails, is composed of three exons, as is the chicken LEAP‐2 homolog (GgLEAP‐2). An analysis of the coding sequence revealed that CjLEAP‐2 is 231 bp long, like GgLEAP‐2, and 93% identical to GgLEAP‐2 at the nucleic acid level. The predicted amino acid sequence of CjLEAP‐2 contained the liver‐expressed antimicrobial peptide 2‐precursor domain and four cysteine residues characteristic of the LEAP‐2 protein. The amino acid sequence of the mature peptide of CjLEAP‐2 was 100% identical to that of GgLEAP‐2. We confirmed that CjLEAP‐2 was transcribed in at least seven tissues, including the digestive system. Additionally, the mature peptide region of CjLEAP‐2 exhibited no polymorphisms in 99 quails from six strains. Taken together, these findings indicate that CjLEAP‐2 is non‐polymorphic and therefore, it likely plays an important role in the innate immunity of quail as it does in chicken.     

Puerperal and parental experiences alter rat preferences for pup odors via changes in the oxytocin system

In the rat, induction of maternal behavior depends on the parity of the female. For example, nulliparous (NP) females need longer exposure to pups than multiparous (MP) or lactating (L) females to exhibit similar maternal behavior. In this study, we investigated the role of brain oxytocin in the approaching behavior of these female rats. Olfactory preferences for pup odors were examined for 8 consecutive days. Each preference test was followed by direct overnight exposure to pups. On the 8th day, MP and L, but not NP females showed robust pup-odor preferences. After the behavioral test, half of the females were exposed to pups for 2 h, whereas the other half were not. The females were then sacrificed to analyze brain oxytocin (OXT) and vasopressin (AVP) activities by cFos immunohistochemistry and to quantify their receptor mRNA expression using real-time PCR. In the paraventricular nucleus (PVN), the percentage of cFos-positive OXT neurons was significantly larger in MP and L females than in NP females after pup exposure. No significant differences were found in cFos expression in OXT neurons of the supraoptic nucleus (SON) or in AVP neurons of either the PVN or SON. Expression of OXT receptor mRNA in the medial preoptic area and amygdala of the control groups was also higher in MP females than in NP females. Finally, we demonstrated that infusion of OXT into the lateral ventricle of NP females promoted preferences for pup odors. These results indicate that puerperal and parental experiences enhance the responsiveness of OXT neurons in the PVN to pup stimuli and establish olfactory preferences for these odors in a parity-dependent manner.     

Feeding territory and variations in behavioural modes of algae‐grazing fish Plecoglossus altivelis ryukyuensis (Ryukyu‐ayu) in subtropical island streams

Abstract – Social and foraging modes in fish often vary between individuals with different body sizes and between populations under different ecological conditions. We studied social and foraging behaviour of algae‐grazing fish Plecoglossus altivelis ryukyuensis (Ryukyu‐ayu) inhabiting Japanese subtropical island streams. Ryukyu‐ayu exhibited four behavioural modes: territorial, schooling, solitary and floating. Their body sizes differed among these modes. Territorial fish predominantly foraged on benthic algae, whereas schooling and solitary individuals frequently consumed drifting materials as well. Schooling fish were smaller than territorial fish, but, unlike some other algae‐grazing fish species, did not use schooling to gain access to food within territories. Territorial fish attacked smaller conspecifics but exhibited lateral display towards larger fish and schooling fish while occasionally attacking grazing gobies. Larger fish maintained larger feeding territories and occupied territories for longer periods than smaller fish did. This suggests that, in Ryukyu‐ayu, intra‐ and interspecific food competitions and relative body size can influence diverse behavioural modes and duration of territory occupation. We also found that Ryukyu‐ayu foraged more often and had larger feeding territories than ayu P. a. altivelis inhabiting temperate streams. We conclude that foraging strategies of Ryukyu‐ayu may have adapted to subtropical island streams, where algal productivity is much lower than that in temperate regions.