Plant species identification using fecal DNAs from red-eared slider and Reeves’ pond turtle in agricultural canals for rural ecosystem conservation

Fecal DNA samples from the red-eared slider and Reeves’ pond turtle, suspected pests of lotus root paddies, were used to identify the plant species eaten by these turtles in order to develop a strategy for rural ecosystem conservation. The fecal samples were obtained from young and adult individuals (mostly female) of both species living in agricultural canals surrounding lotus root paddies in Tokushima Prefecture, Japan. The samples were screened for the presence or absence of DNA from nine plant species using PCR and plant species-specific primers for the rbcL gene of chloroplast DNA. In the red-eared slider, our analysis identified seven plant species in the fecal DNA samples of adults and three plant species in those of young individuals. In Reeves’ pond turtle, our analysis identified two plant species from adult fecal samples and one species from those of young individuals. Thus, adult red-eared sliders consume a greater range of plants than young red-eared sliders or Reeves’ pond turtles. Both turtle species, independently of age, consumed lotus plants and were likely to cause feeding damage to lotus roots. Considering the plant species detected in adult red-eared sliders and these plant habitats, we suggest that this adult turtle is likely to travel between the agricultural canals and the lotus root paddies. These findings will help the development of strategies for preventing damage to lotus roots by these turtles; furthermore, they indicate that fecal DNA analysis will be applicable to investigation of the feeding habits of other animal species.     

Antitumor effect of adriamycin entrapped in liposomes conjugated with anti-bovine tumor antigen monoclonal antibody in leukemic cows

Monoclonal antibody c143 against tumor-associated antigen (TAA) expressed on bovine leukemia cells was conjugated to liposomes containing adriamycin (ADM), and therapeutic effects of conjugates were examined in leukemic or preleukemic cows to prevent the progression of the disease. Five cows with TAA-positive in their peripheral blood lymphocytes were divided into two groups. Each group of cows received 4 injections of ADM alone (0.4 mg/kg) or c143-conjugated liposomes containing the same dose of ADM (L-AMD-c143) through the jugular vein at about 4 day intervals. In three animals treated with L-ADM-c143, the TAA-positive cells gradually decreased with treatment and finally two animals became TAA-negative during a 6 week period and a 14 week period after treatment, respectively. In the control, two animals treated with ADM alone showed only a slight decrease of TAA-positive cells.     

Morphological study on the stomach of the lesser mouse deer (Tragulus javanicus) with special reference to the internal surface.

The stomach of the lesser mouse deer (Tragulus javanicus) was observed macroscopically. It consisted of only three compartments, rumen, reticulum and abomasum without omasum. The rumen was S-shaped with large ventral and caudoventral blind sacs and the reticulum was larger than the abomasum. Internally, the rumen was covered with numerous ruminal papillae even on the pillars and the ruminoreticular fold. These papillae were leaf- or tongue-like shaped and varied in size and density. The reticulum had honey-combed crests and the secondary crests were found rarely. The lips of the reticular groove were prominent and more developed in the aboral part than in the oral one. A sac-like transition zone, which had more prominent mucosal folds than had the floor of the reticular groove, was observed between the caudal end of the reticular groove and the abomasum. Mucosal folds of the abomasum were spiral, low but rather thick. These findings were discussed in view of comparison with other ruminants and of possible functional implications.     

Bovine haptoglobin: single radial immunodiffusion assay of its polymeric forms and dramatic rise in acute-phase sera.

Using purified bovine haptoglobin (Hp) and specific antisera, a single radial immunodiffusion (SRID) assay method has been developed to measure the serum Hp level in cattle. Bovine Hp is a highly polymerized protein showing heterogeneous molecular forms in serum. After treatment with cysteine or glutathione, Hp was partially reduced to a homogeneous form, suitable for SRID assay. This method gives values comparable to those obtained by hemoglobin-binding capacity assay, and has the advantage of being simple and convenient. Although serum Hp was not detectable in healthy cattle, it was found more than 50-fold after invasive surgery, indicating that Hp is a characteristic acute-phase protein in cattle.     

Localization of extracellular matrix receptors in ICGN mice, a strain of mice with hereditary nephrotic syndrome.

Fibrotic degeneration was examined in the kidneys of ICR-derived glomerulonephritis (ICGN) mice, a novel inbred mouse line with a hereditary nephrotic syndrome of unknown etiology considered to be a good model of human idiopathic nephrotic syndrome. In the present study, we histochemically revealed changes in accumulation of extracellular matrix (ECM) components and in localization of integrins, cellular receptors for ECM, in the kidneys of ICGN mice with the progression of renal failure. Excessive accumulation of basement membrane (laminin and collagen IV) and interstitial (type III collagen) ECM components were demonstrated in the glomeruli and tubulointerstitum of ICGN mice. Marked deposition of type I collagen and tenascin was seen only in the glomeruli of ICGN mice but not in those of ICR mice as normal controls. Increased expression of integrin alpha1-, alpha2-, alpha5- and beta1-subunits in glomeruli with fibrotic degeneration and abnormal distribution of alpha6-subunit were noted in the kidneys of ICGN mice. Excessive laminin, a ligand of alpha6beta1-integrin, was demonstrated on the tubular basement membrane, but alpha6-subunit diffusely disappeared on the basal side of the tubular epithelial cells. We presumed that abnormal integrin expression in renal tubules causes epithelial cell detachment, and consequently tubular nephropathy, and results in disorder of ECM metabolism causing excessive accumulation of ECM components in the kidneys of ICGN mice.     

Mycobacterium avium infection through the alimentary tract in mice

Intestinal infection by Mycobacterium avium was investigated in C57BL/6 and BALB/c mouse strains. Single intragastric administration of a massive dose (10(8] or multiple administration of a lower dose (10(7), 10 times) established infection principally in the mesenteric lymph-node (MLN); a continuous or intermittent fecal excretion of the bacilli was detected by 6-8 weeks after the administration. Based on three criteria--isolation of the organisms from the MLN and from feces, and detection of acid-fast bacilli in sections of the MLN--germ-free (GF) BALB/c mice exhibited clearer dose-effect relations than the flora-bearing (FB) counterparts. After intragastric administration, the organisms were probably trapped in the Peyer's patch and then transferred to the MLN at an early period (by 4-7 days), persistent infection thus being established in the MLN. Systemic involvement evolved both in athymic and euthymic mice after a prolonged period of time (more than 40 weeks) showing far more severe involvement in the former regardless of the presence of floral organisms.     

Quantitative capillary reversed passive latex agglutination test for C-reactive protein (CRP) in the dog

A capillary reversed passive latex agglutination test (capillary RPLA) was developed which allows quantification of serum C-reactive protein (CRP) within approximately 15 min. The logarithmic regression line (calibration curve) obtained after measuring each CRP concentration three times in twofold dilutions of a standard canine serum containing 222 g/ml of CRP was y=6.394+0.030x (r=0.995). Capillary RPLA permitted quantification of CRP in the range 6.9–222 g/ml. The coefficients of variation ranged from 10.28% to 12.40%. The recovery rates (percentage recovery) of CRP by capillary RPLA were within the range 87% to 106%. On measuring the CRP concentrations in sera from 78 dogs by capillary RPLA, single radial immunodiffusion (SRID) and enzyme-linked immunosorbent assay (ELISA), close correlations were demonstrated between SRID and capillary RPLA (y=7.250+1.109x, r=0.978), between SRID and ELISA (y=3.042+1.059x, r=0.967), and between capillary RPLA and ELISA (y=1.778+0.929x, r=0.962). Capillary RPLA may be considered useful as a routine biochemical technique for measurement of serum CRP concentration in the dog.Abbreviations CRP C-reactive protein - ELISA enzyme-linked immunosorbent assay - RPLA reversed passive latex agglutination test - SRID single radial immunodiffusion     

Ultrastructural Localization of Hydrogen Peroxide in Host Leaves Treated with AK-toxin I Produced by Alternaria alternata Japanese Pear Pathotype

The rapid generation of reactive oxygen species (ROS), called the oxidative burst, is one of the earliest host responses to pathogen infection or elicitor treatments. Therefore, we looked for the induction of ROS generation in Japanese pear leaves by the host-specific toxin, AK-toxin I using a cytochemical method for detecting H₂O₂. A small amount of non-specific generation of H₂O₂ was found in the cell walls in toxin- and water-treated susceptible and resistant leaves. Thus, the generation of H₂O₂ at cell walls appears to be caused by wounding stress during sampling. Specific generation of ROS, however, was found only in the membrane fragments and extended desmotubules characteristic of modified sites of the plasma membrane in the toxin-treated susceptible leaves. In addition, generation of H₂O₂ at plasma membranes was observed with higher frequency in toxin-treated susceptible leaves. This result indicates that the H₂O₂ generation was associated with damaged sites in the plasmalemma after toxin treatment and perhaps with the formation of membrane fragments from altered portions of the invaginated plasma membrane. Received 21 September 2001/ Accepted in revised form 25 October 2001