Near-infrared spectroscopy evaluated as a technique for estimating udder haemodynamics in the lactating cow

Near-infrared spectroscopy (NIRS) that requires few compact instruments is a non-invasive and continuous method of monitoring haemodynamics in living organs. To assess the feasibility of using NIRS in evaluating the udder haemodynamics in six dairy cows, a probe component consisting of a light-emitting diode and a phototransistor was attached to the left side of the udder surface. Real-time variation of tissue haemodynamics was analysed and the data were transferred to the computer by way of a radiotelemetry system. On induction of temporary congestion by applying pressure with the hand pressed firmly against the left milk vein for 20 s, concentration of oxyhaemoglobin decreased throughout the 20-s duration of congestion, and concentration of deoxyhaemoglobin increased. A surge in blood volume was also observed from the beginning of congestion, then the volume attenuated to baseline. The data demonstrated that the reading seen on the monitor reflects the real-time status of the mammary gland. NIRS shows promise as a useful approach to physiological studies.     

Acute hepatitis in a piglet experimentally inoculated with tissue homogenates from pigs with postweaning multisystemic wasting syndrome

Five 2 day-old colostrum-deprived piglets were inoculated with tissue homogenates from pigs with postweaning multisystemic wasting syndrome. One of the five piglets developed icterus and died 23 days post-inoculation. Histologic examination revealed acute hepatitis. Porcine circovirus type 2 (PCV-2) antigen and nucleic acid were detected in hepatocytes and phagocytic cells. Ultrastructurally, hepatocytes and phagocytic cells had large numbers of cytoplasmic inclusions, which were composed of electron-dense paracrystalline arrays of small non-enveloped viral particles approximately 17 nm in diameter. Apoptotic hepatocytes were confirmed by the TUNEL method and electron microscopic examination. These findings may indicate that hepatocellular necrosis is associated with replication of PCV-2. Apoptosis of hepatocytes also contributes to the pathogenesis of hepatic lesions in this case.     

Lymphocyte subsets and specific IgG antibody levels in clindamycin-treated and untreated dogs experimentally infected with Babesia gibsoni

This study was carried out to clarify the role of lymphocyte subpopulations and Babesia-specific antibody on the treatment of clindamycin in dogs infected with B. gibsoni. Ten beagle dogs were divided into two groups: an untreated group (5 dogs) and a clindamycin-treated group (5 dogs), which was administered clindamycin at 25 mg/ kg body weight, per os, q 12 hr from 7 days to 21 days post-infection (PI). On the acute stage of infection, clindamycin treatment resolved anaemia and other clinical findings. There were no significant differences between treated and untreated dogs either in parasitemia levels or Babesial IgG antibody levels. However, morphological changes that indicated degeneration in the majority of parasites were observed. The numbers of CD4(+) showed a significant increase in treated dogs, especially after treatment. On the chronic stage, CD4(+) cells maintained high level both of the treated and untreated dogs. Although parasitemia maintained low level, their relapses were occurred on the 49th day PI in treated dogs and on the 42nd and 63rd PI in untreated dogs. A rapid humoral antibody response was observed in treated dogs, however, lower humoral antibody responses in untreated dogs after relapses. The antibody levels of treated dogs were significantly higher than those of untreated dogs. These results suggested that clindamycin might not eliminate rapidly parasites from peripheral blood, but damage parasites, which might stimulate efficiently humoral and cellular immunity against Babesia infection, and result in an improvement of clinical conditions.     

Characterization of some Asian isolates of Phytophthora infestans

A total of 401 isolates of Phytophthora infestans were collected from eight Asian regions (Korea, India, Taiwan, Indonesia, Thailand, Nepal, China and Japan) between 1992 and 2000 – 318 from potato and 83 from tomato. The isolates were analysed for mating type, metalaxyl resistance, RG57 fingerprinting, mitochondrial DNA (mtDNA) haplotype and the polymorphism of three allozyme loci, i.e. glucose-6-phosphate isomerase ( Gpi ), peptidase ( Pep ) and malic enzyme ( Me ). The isolates were multilocus-genotyped based on RFLP (RG57) fingerprint, dilocus allozyme genotype, mtDNA haplotype and mating type. Twenty multilocus genotypes were identified among 125 isolates. Of these genotypes, 14 had not been previously reported. Some of the multilocus genotypes were common to isolates from several geographical regions, suggesting migration. The metalaxyl-resistant isolates belonged to the multilocus genotypes JP-1, JP-2, and JP-3. Multilocus genotypes coexisting in a single field were found in following regions: Thailand (1994), central China (1996), Nepal (1997) and Japan (1998 and 2000). The possible origins of certain genotypes are discussed, including the possibility of sexual recombination within the P. infestans populations in Nepal and perhaps Thailand.     

A binding protein for fungal signal molecules in the cell wall of Pisum sativum

In the plant cell wall of Pisum sativum seedlings, we found an NTPase (E.C. 3.6.1.5.) with ATP-hydrolyzing activity that was regulated by an elicitor and suppressors of defense from pea pathogen Mycosphaerella pinodes. The ATPase-rich fraction was purified from pea cell walls by NaCl solubilization, ammonium sulfate precipitation, and chromatography with an ATP-conjugated agarose column and an anion-exchange column. The specific activity of the final ATPase-rich fraction increased 600-fold over that of the initial NaCl-solubilized fraction. The purified ATPase-rich fraction also had peroxidase activity and generated superoxide, both of which were regulated by the M. pinodes elicitor and suppressor (supprescins). Active staining and Western blot analysis also showed that the ATPase was copurified along with peroxidases. In this fraction, a biotinylated elicitor and the supprescins were bound primarily and specifically to ca. 55-kDa protein (CWP-55) with an N-terminal amino acid sequence of QEEISSYAVVFDA. The cDNA clone of CWP-55 contained five ACR domains, which are conserved in the apyrases (NTPases), and the protein is identical to a pea NTPase cDNA (GenBank accession AB071369). Based on these results, we discuss a role for the plant cell wall in recognizing exogenous signal molecules.     

Dietary L-carnitine increases plasma insulin-like growth factor-I concentration in chicks fed a diet with adequate dietary protein level

1. The effect of L-carnitine supplemented into experimental diets with varying dietary protein concentrations (50, 200 and 400 g/kg) on body weight gain and plasma insulin-like growth factor-I (IGF-I) concentration in chicks was examined. 2. Dietary L-carnitine supplementation provided 0, 200, 500 and 1000 mg/kg. Chicks were given the diet ad libitum for 10 d. 3. When L-carnitine was provided as 500 or 1000 mg/kg, body weight gain was significantly improved in birds receiving the 200 and 400 g protein/kg diets. 4. There was an interaction between dietary L-carnitine and protein content on plasma IGF-I concentration. L-carnitine supplementation had little influence on plasma IGF-I concentrations in birds receiving the low protein (50 g/kg) diet. When dietary L-carnitine concentrations were increased from 0 to 1000 mg/kg in the adequate protein (200 g/kg) diet, plasma IGF-I concentrations were also increased. However, when dietary L-carnitine content was more than 500 mg/kg in the 400 g/kg protein group, plasma IGF-I concentration decreased with increasing dietary L-carnitine content. 5. Body weight change correlated significantly with the alteration in plasma IGF-I concentrations in chicks given diets with adequate dietary protein. 6. In conclusion, the improvement in body weight gain caused by dietary L-carnitine supplementation was achieved when chicks were given their dietary protein requirement, which may be partially explained by an increase in plasma IGF-I concentration.     

Demonstration of continuously seropositive population against Borna disease virus in Misaki feral horses,a Japanese strain: a four-year follow-up study from 1998 to 2001

Borna disease virus (BDV)-specific antibodies were monitored in Misaki feral horses annually for 4 years using an electrochemiluminescence immunoassay (ECLIA). Among 130 horses examined, 35 (26.9%) with an ECLIA count above 1000 once or more were judged as BDV seropositive. Throughout the study period, p24 antibodies were more frequent than p40 antibodies in almost all positive animals. Among the 35 seropositive horses, the ECLIA count was consistently high in 12 cases. Eleven horses seroconverted from negative to positive and 7 underwent reversal. The count in the remaining 95 horses (73.1%) remained low for 4 years and these animals were judged as seronegative.     

Inhibition of appressorial adhesion of Pyricularia oryzae to barley leaves by fungicides

The effects of rice blast fungicides known to inhibit melanisation or penetration on the adhesion of Pyricularia oryzae to the leaf surface of barley were investigated. Adhering appressoria were counted after shaking inoculated leaves vigorously with water. On untreated leaves, appressoria began to adhere at the time when appressorial melanisation was initiated. The number of adhering appressoria then increased gradually, and most were melanised. On chlobenthiazone-treated leaves, appressoria were not melanised and were easily detached from the surface. Similar results were obtained with tricyclazole. However, after treatment with tetrachlorophthalide or pentachlorobenzyl alcohol, which are also known to prevent penetration of P. oryzae, melanised appressoria still remained after shaking in water. It is suggested that appressorial melanisation in P. oryzae is involved in appressorial adhesion. The antipenetrant action of the melanin biosynthesis inhibitors chlobenthiazone and tricyclazole may be ascribed to the lack of adhesive intensity necessary to support the mechanical shearing of the cuticle of rice plants.     

Sensitivity to Protectant Fungicides and Pathogenic Fitness of Clonal Lineages of Phytophthora infestans in the United States

ABSTRACT Since 1991, dramatic changes have occurred in the genetic composition of populations of Phytophthora infestans in the United States. Clonal lineages recently introduced into the United States (US-7 and US-8) are more common now than the previously dominant lineage (US-1). To help determine why these changes occurred, four clonal lineages of P. in-festans common during the early 1990s in the United States and Canada were evaluated for sensitivity to the protectant fungicides mancozeb and chlorothalonil using amended agar assays for isolates collected from 1990 to 1994. No isolate or lineage was resistant to either mancozeb or chlorothalonil. There were significant differences among isolates for degree of sensitivity to one fungicide individually, but there were no significant (P = 0.05) differences among the US-1, US-6, US-7, and US-8 clonal lineages for degree of sensitivity to both fungicides. Therefore, resistance to protectant fungicides cannot explain the rapid increase in frequency of the US-7 and US-8 clonal lineages. Three components of pathogenic fitness (latent period, lesion area, and sporulation after 96 h) were tested for the three clonal lineages that were detected most commonly during 1994 (US-1, US-7, and US-8). All but one of the isolates in this analysis were collected during 1994 and evaluated within 10 months of collection by inoculating detached leaflets of the susceptible potato cultivar Norchip. There were significant differences between the US-1 and US-8 clonal lineages for lesion area and sporulation, and between US-1 and US-7 for latent period. The US-6 clonal lineage was excluded from the pathogenic fitness experiments, because no isolates of this lineage were collected during 1994. Compared with US-7 and US-8, US-1 had the longest latent period and the smallest lesions with the least sporulation. Incorporation of the differences between US-1 and US-8 in computer simulation experiments revealed that significantly more protectant fungicide (e.g., 25%) would be required to suppress epidemics caused by the US-8 clonal lineage compared with US-1. These differences in pathogenic fitness components probably contribute to the general predominance of the "new" clonal lineages (especially US-8) relative to the "old" US-1 lineage.