Production of interspecific hybrids between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by ovule culture

Interspecific hybrids were efficiently produced in the cross-incompatible combination between Alstroemeria pelegrina L. var. rosea and A. magenta Bayer by culturing immature ovules with placenta 7–14 days after pollination on 2 g/l Gelrite-solidified MS medium containing 3% (w/v)sucrose. The plants showed intermediate characteristics between the parents and their hybridity was confirmed by karyotype and DNA analyses. The mean number of chromosome association per PMC at metaphase I was 2.60I+6.70II, pollen stainability was20.8%, and they produced viable seeds after self-pollination. Furthermore, mature plants were obtained when the hybrids were backcrossed as male parents with both the parents. The backcross-progeny from A. pelegrina var. rosea × hybrids exhibited 3.8 to 79.7% pollen stainability and that from A. magenta × hybrids 78.8 to 98.3%. Almost all of these plants produced viable seeds after self-pollination, which implies that they can beutilized for breeding of novel cultivars of Alstroemeria. This revised version was published online in July 2006 with corrections to the Cover Date.     

Energy budget of the Japanese flounder Paralichthys olivaceus (Temminck & Schlegel) larvae fed HUFA-enriched and non-enriched Artemia nauplii

The energy budget of the Japanese flounder Paralichthys olivaceus (Temminck & Schlegel) larvae fed enriched (EA) and non‐enriched (NEA) Artemia nauplii was determined by equating energy intake (EI) with the summation of energy channelled to faeces (F), metabolism (M), excretion (U) and growth (G). Larvae (21 days post hatching, 2.2 mg mean wet wt) were reared in six 80‐L circular tanks with three replicates of 160 larvae per tank and fed EA and NEA for 20 days. EI was calculated from the energy content of consumed nauplii, M from the summation of energy for routine, feeding and active metabolisms, U from ammonia excretion and G from energy gained based on weight gain, while F was the difference between EI and the total of other components. The heat increment of larvae was calculated from the difference of O₂ consumption at post‐prandial and routine conditions. Except for G and F, variables were correlated to the dry body weight (W) of larvae in a power function: Y=aW^b. Coefficients a and b were estimated by regression after a logarithmic transformation of the raw data. Overall, growth and survival rates of the larvae fed EA were higher than those fed NEA. For a larval flounder growing from 2 to 20 mg wet wt, the ingested energy was partitioned as follows: 22.8% to faecal loss, 38.3% to metabolism, 1.5% to urinary loss and 37.4% to growth for the EA group, whereas 35.4% to faecal loss, 28.4% to metabolism, 1.3% to urinary loss and 34.9% to growth for the NEA group. Gross conversion and assimilation efficiencies were higher, but the net conversion efficiency was lower in EA‐fed larvae than NEA‐fed larvae. This study suggests that the higher growth and survival rates of the EA‐fed group compared with the NEA‐fed group were attributed to their higher intake of essential fatty acids, higher metabolism and lower energy loss of faeces.     

Genetic differentiation of the bronze featherback Notopterus notopterus between Mekong River and Tonle Sap Lake populations by mitochondrial DNA analysis

ABSTRACT:   Although the bronze featherback Notopterus notopterus is an important fish in South-East Asia, the population structure has not been investigated. In this study, genetic diversity and population structure of the bronze featherback were examined using nucleotide sequence analysis of the mitochondrial DNA control region for 332 fish collected from Tonle Sap Lake and the Mekong River in Cambodia. The average nucleotide diversity (π) of all samples was 0.033, and the Mekong River samples had higher nucleotide diversity (0.034) than Tonle Sap Lake samples (0.028). The nucleotide diversity between the lake and river samples varied from 0.029 to 0.037. The genetic differentiation between the river and lake populations was also supported by the pairwise F -statistic values and hierarchical analysis of molecular variance, indicating that the Tonle Sap Lake population is genetically isolated from the population in main stream of the Mekong River.     

Vitamin C requirement of kuruma shrimp postlarvae, Marsupenaeus japonicus (Bate), using l-ascorbyl-2-monophosphate-Na/Ca

l ‐ascorbyl‐2‐monophosphate‐Na/Ca (AMP‐Na/Ca) was used as a vitamin C source to investigate the ascorbic acid (AsA) requirements on growth performance and stress resistance of the post‐larval kuruma shrimp, Marsupenaeus japonicus. Purified carrageenan‐microbound diets with six levels of AMP‐Na/Ca, AsA equivalent to 0, 20, 56, 87, 759 and 1697 mg kg^?1 diet were fed to triplicate groups of M. japonicus (mean initial weight 16±0.3 mg) for 30 days. The diets with AsA 0, 20 and 56 mg kg^?1 showed high cumulative mortality after 10 days of feeding. After the 30‐day trial, the shrimp fed these diets had significantly lower survival and weight gain (WG, %) than those fed the 87, 759 and 1697 mg AsA kg^?1 diets. Specific growth rate and individual dry weight showed the same pattern as WG (%). There were no significant differences in growth performance among the groups fed the AsA levels at 87, 759 and 1697 mg kg^?1 at the termination of feeding trial. Broken‐line regression analysis indicated that 91.8 mg AsA kg^?1 in the diet was the optimum for post‐larval shrimp. On the other hand, dietary level of more than 800 mg AsA kg^?1 was needed to ensure high resistance to stressful conditions such as osmotic and formalin stressors.     

Influence of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder, Paralichthys olivaceus

A feeding trial using five semi-purified diets (50% crude protein) was conducted to investigate the effects of different dietary amino acid patterns on growth and body composition of juvenile Japanese flounder. The control diet contained casein and gelatin as intact protein sources and four other diets contained 30% casein–gelatin (2:1, w/w) and 20% crystalline amino acids (CAA). CAA were added to the diets to simulate the amino acid pattern found in red sea bream egg protein (REP), Japanese flounder larvae whole body protein (FLP), Japanese flounder juvenile whole body protein (FJP), and brown fish meal protein (BFP), respectively. The test diets were fed to triplicate groups of juveniles (2.75±0.05 g) twice a day for 40 days to evaluate weight gain, survival, feed conversion efficiency (FCE), protein efficiency ratio (PER), and apparent protein utilization (APU). The apparent retention of total dietary amino acids in the whole body and A/E ratios of the whole body were also evaluated. The highest weight gain was observed in fish fed the diet containing the dietary amino acid pattern of BFP followed by fish fed the control, FJP, FLP and the REP diets. Percent survival, FCE, PER and APU were also significantly (P<0.05) affected by the amino acid pattern in the diets, indicating the highest value in fish fed the BFP diet. Except for a few amino acids, the amino acid composition of the whole body did not show marked differences with different dietary amino acid pattern. Results suggest that BFP could be more suitable as a reference amino acid pattern in the diet of juvenile Japanese flounder compared to the amino acid pattern of FLP, FJP or REP.     

Lactic acid bacteria effective for regulating the growth of contaminant bacteria during the fermentation of <Emphasis Type="Italic">Undaria pinnatifida</Emphasis> (Phaeophyta)

ABSTRACT:   Lactic acid fermentation of seaweed is a recent topic and quite limited information is available on culture conditions. To know the suitable strains for use as a starter culture for seaweed fermentation, 14 lactic acid bacteria (LAB) strains, including 11 species, were tested in culture conditions prepared with or without salt. A commercial product of Undaria pinnatifida powder was used as a substrate for fermentation without sterilizing. Starter-suitability of the LAB strains was assessed from their predominance after culture. Among the tested strains, Lactobacillus brevis, Lactobacillus plantarum, Lactobacillus casei and Lactobacillus rhamnosus showed high (>90%) predominance in their cultures, while control cultures prepared without inoculation of LAB did not show any detectable growth of acid producing bacteria and spoiled. A total of 102 strains not showing acid producing activity were isolated and characterized from spoiled cultures, and all the isolates were observed as Bacillus strains, including 64 strains (62.7%) and 16 strains (15.7%) of a Bacillus cereu s-related and B. fusiformis -related species, respectively. The Undaria substrate before fermentation contained culturable microorganisms at 1.4–3.1 × 10² CFU/g, but the Bacillus cereu s-related strain was not a major composition, suggesting a concern of selective growth of the Bacillus cereu s-related strain during the spoiled fermentation.     

Midgut juice of Plutellaxylostella highly resistant to Bacillusthuringiensis Cry1Ac contains a three times larger amount of glucosinolate sulfatase which binds to Cry1Ac compared to that of susceptible strain

Midgut juice of Plutellaxylostella strain PXR which is resistant to Cry1Ac was biochemically characterized relative to the susceptible PXS strain. The midgut juice of PXR (PXR-Juice) was shown to process Cry1Ac protoxin to 60 kDa active toxin with the same processing pattern as that of juice from PXS (PXS-Juice) in SDS–PAGE. PXS larvae which were given the Cry1Ac toxin pre-processed with PXR-Juice were killed with the same rate as that with Cry1Ac pre-activated by trypsin. PXR-Juice was found to contain three times larger amount of 66 kDa protein (P66) than PXS-Juice and the N-terminal amino acid sequence of P66 was matched to that of glucosinolate sulfatase in data base search. The protein band of P66 was coincided with the band of p-nitro phenyl sulfatase activity in zymogram. P66 purified to homogeneity in SDS-PAGE bound to Cry1Ac and soybean agglutinin, and K_D for Cry1Ac was estimated to be 718 nM with surface plasmon resonance analysis. Using purified sulfatase, K_m and V_max were estimated and involvement of the enzyme in the PXR resistance was discussed.     

Identification of cancer stem cells derived from a canine lung adenocarcinoma cell line

Accumulating evidence supporting the cancer stem cell (CSC) hypothesis is based on the finding that tumors contain a small population of self-renewing cells that generate differentiated progeny and thereby contribute to tumor heterogeneity. CSCs are reported to exist in several human cancers, yet only a few reports demonstrate the existence of CSCs in primary lung cancer in dogs. In this study, the authors established a cancer cell line derived from a canine primary lung adenocarcinoma and identified a side population (SP) of cells that displayed drug-resistant features. To confirm the characteristics of these SP cells, the authors investigated the tumorigenicity of the cells in vivo by using a nude mouse xenograft model. Only 100 SP cells were able to give rise to new tumors, giving a 10-fold enrichment over the main population (MP) of cells, suggesting that these cells have the cancer-initiating ability of CSCs. Further studies characterizing CSCs in canine lung adenocarcinoma might contribute to the elucidation of the mechanisms of tumorigenesis and to the establishment of novel therapeutic strategies.     

Echocardiographic estimation of left atrial pressure in beagle dogs with experimentally-induced mitral valve regurgitation

Non-invasive and immediate estimation of left atrial pressure (LAP) is very useful for the management of mitral regurgitation (MR), and many reports have assessed echocardiographic estimations of LAP to date. However, it has been unclear of which examination and evaluate article possess the best accuracy for the MR severity. The present research aims to establish the echocardiographic estimation equation of LAP that is well applicable for clinical MR dogs. After the chordae tendineae rupture was experimentally induced via left atriotomy in six healthy beagle dogs (three males and three females, two years old, weighing between 9.8 to 12.8 kg), a radio telemetry transmitter catheter was inserted, which allows the continuous recordings of LAP without the use of sedation. Approximately 5 weeks after the surgery, echocardiographic examination, indirect blood pressure measurement, measurement of plasma atrial natriuretic peptide (ANP) and LAP measurement by way of the radio telemetry system was performed simultaneously. Subsequently, simple linear regression equations between LAP and each variable were obtained, and the equations were evaluated whether to be applicable for clinical MR dogs. As a result, the ratio of early diastolic mitral flow to early diastolic lateral mitral annulus velocity (E/Ea) had the strongest correlation as maximum LAP=7.03*(E/Ea)-54.86 (r=0.74), and as mean LAP=4.94*(E/Ea)-40.37 (r=0.70) among the all variables. Therefore, these two equations associated with E/Ea should bring more precise and instant estimations of maximum and mean LAP in clinical MR dogs.     

Acute myeloid leukemia with mastocytic and megakaryocytic differentiation in a calf

A diagnosis of acute myeloid leukemia was made in a 10-month-old Holstein female calf. The leukemia was macroscopically characterized by great enlargement of the spleen and moderate enlargement of some lymph nodes. Histochemical and immunohistochemical examination disclosed the presence of neoplastic cells either containing metachromatic and tryptase-positive granules or expressing factor VIII-related antigen. The granules, which were positive for naphthol AS-D chloroacetate esterase and did not have particulate contents, were distinct from those of basophilic leukemia cells. This leukemia was thought to be derived from a common myeloid progenitor capable of giving rise to megakaryocyte-erythrocyte progenitors and granulocyte-monocyte progenitors with the ability to differentiate into mast cells.