Cranial morphological homogeneity in two subspecies of water deer in China and Korea

The water deer (Hydropotes inermis) has conventionally been classified into two subspecies according to geographic distribution and pelage color pattern: H. i. inermis from China and H. i. argyropus from Korea. However, the results of a recent molecular study have called this into question. To further reappraise this classification, we examined morphological variation in craniodental measurements of these 2 subspecies. Results of univariate and multivariate analyses demonstrated that these 2 subspecies are not well-differentiated, suggesting that individuals of the 2 populations share common morphological traits. Despite the distribution of the subspecies at different latitudes, no clear morphocline was detected, suggesting that Bergmann’s rule does not apply in this case. Discriminant analysis indicated that the characteristics of individuals are shared by both populations, suggesting that not all individuals can be assigned to their original population. Results of principal component analysis showed that the two populations shared more than 75% of individuals, congruent with the “75% rule” of subspecies classification. In both the neighbor-joining and unweighted pair group methods with arithmetic mean cluster analyses, specimens of H. i. argyropus and H. i. inermis were highly mixed within the cladograms. These results suggest that the overall morphological variation in the 2 subspecies overlaps considerably and that there is no coherent craniofacial difference between the 2 groups. The present findings combined with prior observations from molecular biogeography point out that the taxonomic division of water deer into 2 subspecies should be revisited.     

Analysis of the equine ovarian structure during the first twelve months of life by three-dimensional internal structure microscopy

A three-dimensional internal structure microscopy (3D-ISM) can clarify the anatomical arrangement of internal structures of equine ovaries. In this study, morphological changes of the equine ovary over the first 12 months of life were investigated by 3D-ISM in 59 fillies and by histological analysis in 2 fillies. The weight and volume of the paired ovaries initially decreased from 0 to 1 months to 2 to 3 months of age and then significantly increased at 8 to 12 months of age. The ovulation fossa was first observed around the 3rd month and became evident after the 6th month. The number of follicles with a diameter of ≥10 mm and the diameter of the largest follicle increased gradually after 6 months of age. On a volume basis, the medulla accounted for nearly 90% of the whole ovary at 0 to 1 months of age, but significantly decreased from 2 to 3 months of age. The volume of the cortex increased progressively after birth and reached approximately 60% of the total volume at 8 to 12 months of age. This significant development of the cortex coincided with the increased number and size of large follicles observed from 6 months of age. These results suggest that the development of the cortex plays a role in the maturation of the follicles and the equine ovary undergoes substantial morphological changes postnatally until puberty.     

Glycophorin A-knockout mice, which lost sialoglycoproteins from the red blood cell membrane, are resistant to lethal infection of Babesia rodhaini

Recent in vitro-based studies using several Babesia spp. have suggested that sialic acids and/or sialoglycoproteins on host red blood cells (RBCs) play an important role in their invasion of RBCs. In the present study, we analyzed the RBC characteristics of glycophorin A (GPA)-knockout mice and studied their in vivo susceptibility to lethal infection of Babesia rodhaini for the first time. In immunoblot and lectin blot analyses, glycoproteins containing O-linked oligosaccharides terminated with alpha2-3-linked sialic acids disappeared from the RBCs of GPA homozygous ((-/-)) mice. Flow cytometric analysis showed a remarkable reduction of Maackia amurensis lectin II binding to the surface of GPA(-/-) RBCs relative to control RBCs, indicating an appreciable loss of alpha2-3-linked sialic acids on the RBC surface of GPA(-/-) mice. Importantly, while B. rodhaini caused lethal infection in wild-type mice, the infected GPA(-/-) mice showed inhibition of parasite growth and eventually survived. These results indicate that RBC sialoglycoproteins lost in GPA(-/-) mice are involved in the in vivo growth of B. rodhaini, probably functioning as essential molecule(s) for the parasite invasion of host RBCs in the blood circulation.     

Development of loop-mediated isothermal amplification (LAMP) method for diagnosis of equine piroplasmosis

Loop-mediated isothermal amplification (LAMP) is a novel nucleic acid method whereby DNA is amplified with high specificity, efficiency, and rapidity under isothermal conditions using a set of four specifically designed primers and a DNA polymerase with strand displacement activity. In this study, we used LAMP primer sets designed from EMA-1 and Bc 48 genes for detection of Theileria equi and Babesia caballi infections, respectively. These primer sets specifically amplified DNA of the respective parasites. Both primer sets amplified T. equi and B. caballi up to 10(-6) dilution of 10-fold serially diluted samples. Furthermore, DNA extracted from blood collected from a horse experimentally infected with T. equi was amplified by a T. equi LAMP primer set from days 2 to 35 post-infection, demonstrating the high sensitivity of these primers. Of 55 samples collected from China, 81.8% and 56.3% were positively detected by LAMP for T. equi and B. caballi infections, respectively. In contrast, 91.8% and 45.9% of the 37 samples collected from South Africa were LAMP positive for T. equi and B. caballi, respectively. These results suggest that LAMP could be a potential diagnostic tool for epidemiological studies of equine piroplasmosis.     

An outbreak of teat papillomatosis in cattle caused by bovine papilloma virus (BPV) type 6 and unclassified BPVs

Out of 700 heifers at a local farm in Hokkaido, the Northern island of Japan, 560 (80%) were found to have benign teat tumors. All of the analyzed tumors were macroscopically of the flat-and-round type, and no other types such as rice-grain or frond epithelial type were found. The lesions were characterized by epithelial hyperplasia, acanthosis and hyperkeratosis. Unlike in typical fibropapilloma, fibroplasia of the underlying dermis was not observed. Bovine papilloma virus (BPV) capsid antigen and virus particles were found in basophilic intranuclear inclusions of the stratum granulosum of the epidermis by immunohistochemistry and electron microscopy, respectively. BPV-specific DNA was also detected in the lesions. By means of the polymerase chain reaction (PCR) and DNA sequencing of the PCR products, the viruses causing this outbreak were identified mainly as BPV-6 (64%), partly as unclassified BPVs (14%) and their co-infections (21%). Our findings suggest that this outbreak of benign teat tumors was associated with several BPV types.     

Dimethoxypyrimidines as Novel Herbicides. Part 1. Synthesis and Herbicidal Activity of Dimethoxyphenoxyphenoxypyrimidines and Analogues

A number of 6-(4-phenoxyphenoxy)pyrimidines and triazines were synthesized and their herbicidal activity was measured. Compounds with the methoxy groups at the 2- and 4-positions on the pyrimidine and triazine rings exhibited high herbicidal activity. Introduction of a substituent into the 5-position of the pyrimidine ring diminished the activity. In the phenoxyphenoxy substructure at the 6-position, the central ether bond can be replaced by a methylene group without loss of activity. The optimum substituent on the terminal phenyl ring was 3-trifluoromethyl. The compounds showed a strong Hill reaction inhibition, but unfortunately showed poor selectivity between weeds and crops.     

Effects of double harvesting on estimated total digestible nutrient yield of forage rice

We examined the effects of cultivar (Taporuri, Mohretsu, Tachiaoba, and Hinohikari) on estimated total digestible nutrient (TDN) yield of forage rice in double harvesting. The total estimated whole-plant TDN yield (i.e., of the first crop plus second crop) of Taporuri was the highest in double harvesting. In the first crop, the estimated whole-plant TDN yield of Taporuri was the highest. In the second crop, the estimated whole-plant TDN yield of Taporuri was almost the same as those of Mohretsu and Tachiaoba but higher than that of Hinohikari. We also examined the effects of cultivation method (double harvesting or single harvesting) on estimated TDN yield of forage rice. In Taporuri and Mohretsu, the total estimated whole-plant TDN yield was similar in double harvesting and single harvesting. However, in Tachiaoba and Hinohikari, it was lower in double harvesting than in single harvesting. It is important to maximize the amount of nutrition in leaf and stem rather than panicle because the grain is not digested well by cattle. In the double harvesting, the total estimated leaf blade (leaf) TDN yield of Taporuri was higher than that of Hinohikari and almost the same as those of Tachiaoba and Mohretsu. The total estimated leaf sheath plus stem (stem) TDN yield of Taporuri was the highest. In comparison of the total TDN yield of double harvesting with that of single harvesting, the total estimated leaf and stem TDN yields were higher in double harvesting than in single harvesting in all cultivars except for Hinohikari. Thus, double harvesting of Taporuri is an effective way of reducing the overall loss of nutrition from the crop.     

Utilization of genetically modified soybean meal in Nile tilapia <Emphasis Type="Italic">Oreochromis niloticus</Emphasis> diets

The utilization of genetically modified soybean meal (GM SBM) was compared with that of non-GM SBM in Nile tilapia. Four experimental diets were formulated to include either non-GM or GM SBM at 34 or 48%, respectively. These diets were fed to juvenile Nile tilapia (49.5 g average weight) for 12 weeks. The uptake of the cauliflower mosaic virus 35S promoter fragment of the GM SBM in fish muscle was examined at 8th and 12th week. After 12th week, fish were fed the non-GM SBM diets to determine the residual span of the incorporated promoter fragment. There was no significant difference in specific growth rate or feed efficiency between GM and non-GM groups at the same inclusion level. A small number of muscles from fish receiving both levels of GM SBM diet were positive for the promoter fragment. Additionally, the promoter fragment was not detected by the second day after changing to the non-GM SBM diets. These results indicate that the utilization of GM SBM was similar to that of non-GM SBM and the promoter fragment was rarely found in fish muscles, suggesting that suitability and safety of GM SBM in Nile tilapia diet were similar to those of non-GM SBM.     

Isolation and molecular characterization of hemocyte sub-populations in kuruma shrimp Marsupenaeus japonicus

Fisheries Science - Crustacean hemocytes, which have usually been classified morphologically based on methods using Giemsa or May-Giemsa stains, have recently been categorized using monoclonal...     

Possibility of anisakid larvae infection in farmed salmon

SUMMARY: Nematodes belonging to the family Anisakidae including Anisakis simplex and Pseudoterranova decipiens are known to cause anisakiasis when their live larvae are ingested by humans. We estimated the possibility of anisakid infection to salmonids, farmed in sea net-pens at Onagawa Bay, Miyagi, Japan, in 1992, 1998 and 1999, by direct examination of the edible muscle and examination of the contents of the alimentary canal. From direct examination of the muscle, no nematode was found in the 249 farmed coho salmon Oncorhynchus kisutch and 40 farmed rainbow trout Oncorhynchus mykiss . In contrast, third-stage larvae of Anisakis simplex were found in seven of 14 wild coho salmon caught in Russia and all the 40 wild chum salmon Oncorhynchus keta caught at Nemuro and Kesennuma in 1998. The stomach and intestines of 521 farmed coho salmon and 40 farmed rainbow trout were examined carefully for the existence of possible carrier organisms such as crustaceans, fish or squid. Such carrier organisms were not found in the stomach and intestines of farmed fish. Thus, we conclude that the possibility of anisakid infection is very low in farmed salmonids.