Accessory corpora lutea formation in pregnant Hokkaido sika deer (Cervus nippon yesoensis) investigated by examination of ovarian dynamics and steroid hormone concentrations

Generally, sika deer conceive a single fetus, but approximately 80% of pregnant females have two corpora lutea (CLs). The function of the accessory CL (ACL) is unknown; moreover, the process of ACL formation is unclear, and understanding this is necessary to know its role. To elucidate the process of ACL formation, the ovarian dynamics of six adult Hokkaido sika deer females were examined ultrasonographically together with peripheral estradiol-17β and progesterone concentrations. ACLs formed in three females that conceived at the first estrus of the breeding season, but not in those females that conceived at the second estrus. After copulation, postconception ovulation of the dominant follicle of the first wave is induced by an increase in estradiol-17β, which leads to formation of an ACL. A relatively low concentration of progesterone after the first estrus of the breeding season is considered to be responsible for the increase in estradiol-17β after copulation.     

Positional strategy of trunk muscles among aquatic,semi-aquatic and terrestrial species in Urodela

Clarification of the trunk structure in Urodela is important in understanding the locomotive evolution of basal tetrapods. The components of the muscular trunk wall among Urodela using different modes of locomotion were compared. Since the whole trunk may be used for swimming and the effect of limbs may be small in the more aquatic species, they showed smaller differences in the trunk muscles among anterior, middle and posterior sections of the trunk. By contrast, in the more terrestrial species, the dorsal and abdominal muscles are larger in the middle section than those in the anterior and posterior sections. High compressive stresses occur in the supporting limbs and their insertion at the trunk on the ventral side, and spread from the forelimbs along the back to the supporting hindlimbs on the dorsal side. Tensile stresses occur in the middle ventral part. The components of the trunk muscles among the three sections may reflect differences in stresses occurring in the trunk of the more terrestrial species. The findings also suggest that in the middle section, larger dorsal muscles for stiffening the back to maintain posture and larger abdominal muscles are responsible for balancing the body weight while it is supported by the limbs in the more terrestrial species.     

Comparative functional anatomy of hindlimb muscles and bones with reference to aquatic adaptation of the sea otter

Although the sea otter (Enhydra lutris) is a complete aquatic species, spending its entire life in the ocean, it has been considered morphologically to be a semi-aquatic animal. This study aimed to clarify the unique hindlimb morphology and functional adaptations of E. lutris in comparison to other Mustelidae species. We compared muscle mass and bone measurements of five Mustelidae species: the sea otter, Eurasian river otter (Lutra lutra), American mink (Neovison vison), Japanese weasel (Mustela itatsi) and Siberian weasel (M. sibirica). In comparison with the other 4 species, E. lutris possessed significantly larger gluteus, popliteus and peroneus muscles, but smaller adductor and ischiopubic muscles. The popliteus muscle may act as a medial rotator of the crus, and the peroneus muscle may act as an abductor of the fifth toe and/or the pronator of the foot. The bundles of the gluteus superficialis muscle of E. lutris were fused with those of the tensor fasciae latae muscle and gluteofemoralis muscles, and they may play a role in femur abduction. These results suggest that E. lutris uses the abducted femur, medially rotated crus, eversion of the ankle and abducted fifth digit or extended interdigital web as a powerful propulsion generator. Therefore, we conclude that E. lutris is a complete aquatic animal, possessing differences in the proportions of the hindlimb muscles compared with those in other semi-aquatic and terrestrial mustelids.     

Dual effects of reversed winter–spring temperatures on year‐to‐year variation in the recruitment of chub mackerel (Scomber japonicus)

To clarify the effects of temperature on the recruitment of chub mackerel (Scomber japonicus) in the North Pacific, we investigated the influence of winter surface temperature (WST) on spawners at the time of maturity around the spawning grounds and the influence of ambient spring temperature on larvae using estimated temperature (ET) obtained from particle tracking experiments. We found a significant positive correlation between ET approximately 10 days following hatching and the recruitment per spawning stock biomass (RPS) after 2000. The closer (more meandering) the Kuroshio Current (KC) axis was in relation to the spawning ground, the higher (lower) the spring surface temperature and the higher (lower) RPS was in the spawning ground. In contrast, WST inside KC near the maturity/spawning ground was significantly negatively correlated with RPS. A significant negative correlation between the temperatures in winter and spring was detected in the area after 2000, when the conditions of the Pacific decadal oscillation index and the stability of the Kuroshio Extension were synchronous, indicating that KC shifted northward during this time. The reversed temperature pattern was consistent with the winter–spring movement of KC axis in the offshore direction and was correlated with the winter–spring difference in the intensity of the Aleutian low. These results suggest that the annual variation in chub mackerel recruitment after 2000 was strongly affected by the combined effects of ambient temperature because of the reversal of conditions that occurred between winter and spring around the maturity/spawning ground, which was related to the KC path.     

Sodium alginate supplementation modulates gut microbiota,health parameters,growth performance and growth‐related gene expression in Malaysian Mahseer Tor tambroides

This study investigated the effects of sodium alginate supplementation on gut microbiota composition, health parameters, growth performances and growth‐related gene expression of Malaysian mahseer. Five test diets were formulated by supplementing 0%, 0.1%, 0.2%, 0.4% and 0.8% sodium alginate. Triplicate groups of juvenile Tor tambroides (2.19 ± 0.05 g) were stocked in 15 aquaria (20 individuals per aquarium) and fed at 3.0% body weight per day for 60 days. PCoA and UPGMA analysis showed that gut bacterial community were more convergence in higher sodium alginate‐supplemented diets. The percentage of Porphyromonadaceae, Bacteroides, Plesiomonas and Shewanella were substantially higher and Aeromonas, Entomoplasmatales and Prevotellaceae were drastically lower in higher sodium alginate (0.2%–0.8%) diets. Sodium alginate supplementation (≥0.2%) significantly improved the haematocrit value and respiratory burst activity of T. tambroides. Growth performances and feed utilization were significantly higher in 0.2%–0.4% sodium alginate‐supplemented diets. The increased growth rate of T. tambroides was governed by both hyperplastic and hypertrophic muscle growth. Real‐time PCR data demonstrated that most of the growth‐related genes were significantly upregulated in 0.2%–0.4% sodium alginate‐supplemented diet. Finally, it can be concluded that sodium alginate should be supplemented at 2 g/kg in practical fish feed formulation.     

Census and effective population sizes of white‐spotted charr (Salvelinus leucomaenis) in a fragmented landscape

Several habitat models have been proposed to predict population size for stream fishes and to guide habitat assessment and monitoring techniques. However, most models do not incorporate the potential advantage of molecular genetic markers. We conducted a field survey and microsatellite DNA analyses to quantify the relationships among genetic diversity, census/effective population size and habitat variables in fragmented populations of white‐spotted charr (Salvelinus leucomaenis). The census population size significantly increased with the stream length, the number of pools and a pool‐riffle sequence index, a proxy for channel‐unit habitat type complexity within reaches. Population density was correlated with the pool‐riffle sequence index only. Genetic diversity and effective population size were not correlated with the habitat variables or census population size. There was a lack of isolation‐by‐distance population structure in the studied populations. Our results suggest that stream length and the number of pools within reaches associated with habitat complexity are the habitat variables that explain the majority of variation in population size of white‐spotted charr. Our findings provide further evidence that census population size per se is a poor indicator of the inclusive genetic diversity within populations in a fragmented landscape.     

Spectrophotometrical characteristics in the near infrared region in beech (<Emphasis Type="Italic">Fagus crenata</Emphasis>) and pine (<Emphasis Type="Italic">Pinus densiflora</Emphasis>) litters at the various decomposing stages

We examined the relationships between the absorptional characteristics in the near infrared region and the chemical changes of decomposing beech (Fagus crenata) and pine (Pinus densiflora) litters. Spectra as well as the concentrations of chemical substances approached each other and converged with decomposition, although both initial characteristics differed markedly between beech and pine. This indicated that the fundamental chemical structures were almost the same, although their organochemical composition differed. Specific absorption bands for lignin, polysaccharide, and protein were identified at 2,140 and 1,670 nm, 2,270, 1,720, 1,590, and 1,216 nm, and 2,350 nm, respectively. Absorbance at 1,670 nm, peculiar band of aromatics, showed a positive correlation with lignin concentration, which suggested the relative increment of aromatics due to condensed lignin in decomposing litters. Absorbance at 2,140 nm, characterized as the C–H bond in HRC = CHR, showed a negative correlation with lignin concentration, which suggested the decrements of some structures such as side-chains in lignin polymers unrelated to aromatics. Absorbance at 2,270, 1,720, and 1,216 nm, specified to O–H/C–O/C–H bonds in saccharide, might reflect the change of polysaccharide during decomposition because they showed a positive correlation to polysaccharide concentration. In the same way, absorbance at 2,350 nm, identified to the C–H/CH₂ bonds in protein, showed a negative correlation to nitrogen concentration in decomposing litters, which might indicate that the C–H/CH₂ bonds in protein decreased with decomposition due to microbial consumption of carbon in protein. Our findings suggested the possibility that the spectral changes indicate the litter digestibility during decomposition and that also explain the compositional change in decomposing litters.     

Effects of polyline simplification of dynamic GPS data under forest canopy on area and perimeter estimations

The objectives of this study were to determine Global Positioning System (GPS) positional errors while moving under the forest canopy and to clarify the effects of polyline simplification on area and perimeter estimations. We used the Pathfinder Pro XR and GPSMAP 76S, which are categorized as “high-end mapping” and “general navigation” GPS receivers, respectively. The field tests were conducted in both natural and plantation forests. The results showed that the Pathfinder Pro XR, which has better multipath rejection technology, worked well, especially in the plantation forest under unfavorable conditions of higher stand density. We used analysis of variance to clarify the effects of the receiver type, positioning mode, stand type, and polyline simplification method on area and perimeter estimations. The receiver type and positioning mode were found to be significant factors that affected area estimation. The Pathfinder Pro XR estimated the area more accurately than the GPSMAP 76S, and differential GPS estimated the area more accurately than autonomous GPS. With respect to the perimeter, the receiver type, positioning mode, and polyline simplification method were found to be significant factors. The results showed that perimeter estimation was improved by using the velocity filter, and further improved by using the velocity filter and Douglas-Peucker algorithm, especially when the Pathfinder Pro XR was used. The GPSMAP 76S estimated the perimeter accurately without any filtering because its default speed filter worked well, even though the GPSMAP 76S is a general navigation GPS receiver.     

Detection of the EGFP sequence in breast muscle of 3‐year‐old chicken after transfection using sonoporation

In our continuing effort to generate transgenic chickens, sonoporation was chosen to insert an exogenous gene into the chicken genome. An EGFP expression vector (pCAG‐EGFPac) and microbubbles were injected into the central disc of stage‐X blastoderm or the germinal crescent of stage‐4 embryos, followed by ultrasonic vibration. Nineteen chicks out of 108 treated embryos hatched, six females and six males out of these 19 chicks grew to sexual maturity and two females and three males lived for 3 years. Genomic DNA from 17 out of 35 gonads from embryos and chicks that died before sexual maturity was EGFP‐positive by PCR. No EGFP sequence was detected in the genomic DNA of 322 embryos from six sexually mature females and the semen from four sexually mature males by PCR. When genomic DNA was obtained from various tissues of five 3‐year‐old chickens, the EGFP sequence was amplified from the genomic DNA of the breast muscle of a female (No. 85). The above sequence was subjected to DNA sequencing and verified to be the EGFP sequence. These results showed that sonoporation is an effective tool for the transduction of exogenous genes into chicken embryos for the generation of transgenic chickens.     

Ecology of Armillaria species on conifers in Japan

Distribution, host preference and pathogenicity of Japanese Armillaria species on conifers were investigated on the basis of field collections of 65 isolates. We identified seven Armillaria species from 19 conifer species including six major Japanese plantation conifers using mating tests and sequences of the translation elongation‐1 α gene. Armillaria mellea, Armillaria ostoyae, Armillaria cepistipes and Armillaria sinapina were frequently collected, whereas Armillaria nabsnona, Armillaria tabescens and a biological species Nagasawa’s E were rare. On the basis of host condition when the isolates were collected, A. mellea, A. ostoyae, A. cepistipes and A. tabescens are considered as moderate to aggressive pathogens of conifers in Japan.