Oxidative dimers produced from protocatechuic and gallic esters in the DPPH radical scavenging reaction

DPPH (2,2-diphenyl-1-picrylhydrazyl) radical scavenging reactions of protocatechuic and gallic acids, and their methyl esters, have been investigated by NMR. In acetone, methyl protocatechuate was gradually converted to a Diels-Alder adduct of two molecules of the intermediate quinone in the reaction with DPPH radical, whereas methyl gallate rapidly gave a symmetrical dimer via a putative quinone precursor. Both dimers are rather unstable and their structures have been deduced by in situ NMR measurements of the reaction mixtures. Gallic acid also gave a corresponding symmetrical dimer in the same reaction as methyl gallate, although protocatechuquinone produced from protocatechuic acid did not yield a Diels-Alder adduct, unlike its methyl ester. Interestingly, these dimer formations were not observed in methanol solution.     

COMPARATIVE EVALUATION OF HYDROPIC DEGENERATION OF THE LIVER IN DAIRY CATTLE THROUGH BIOCHEMISTRY, ULTRASONOGRAPHY AND DIGITAL ANALYSIS

Ultrasonography of the liver of 181 Holstein-Friesian cows was performed and blood samples were collected for analysis. The hepatic ultrasonograms were evaluated and the echoes were analyzed digitally. After slaughter, liver specimens were taken and examined histopathologically. Of the 181 animals, 120 had a normal liver and 61 had hydropic degeneration of the liver, diagnosed through histopathologic examination. Diagnostic accuracy rates for hydropic degeneration were determined based on the following test positive conditions: a) for biochemical analysis—high levels of aspartate aminotransferase, alanine aminotransferase, total bilirubin and non-esterified fatty acids; b) for ultrasonography—presence of dark pattern and blurring of edges; and c) for digital analysis—low echo means at 1 cm and 3 cm from the hepatic surface. Digital analysis had the highest overall specificity, accuracy and positive predictive values for hydropic degeneration, followed by ultrasonography. The results suggest that ultrasonography and digital analysis of hepatic ultrasonograms can be used for diagnosis of hydropic degeneration of the liver in place of biochemical analysis.     

EVALUATION OF FATTY INFILTRATION OF THE LIVER IN DAIRY CATTLE THROUGH DIGITAL ANALYSIS OF HEPATIC ULTRASONOGRAMS

Ultrasonography of the liver of 49 Holstein-Friesian cows was performed, liver specimens were taken, examined microscopically and the fatty occupying rate (FOR) was calculated. Echoes from the hepatic B-mode ultrasonograms were quantified as histogram mean (Emean) and histogram mode (Emode) of echo amplitudes within various areas at a depth of 1–9 cm from the hepatic surface. Of the 49 animals, 26 had a normal liver and 23 had fatty infiltration of the liver, diagnosed through histopathological examination. Fatty occupying rate ranged from 1.7 to 64.5%, with 11 animals having 1–15% FOR (mild fatty infiltration), 6 having 15.1–30% FOR (moderate fatty infiltration) and 6 having > 30% FOR (severe fatty infiltration). At 1 cm, severe fatty infiltration had higher Emean and Emode than normal liver (p < 0.05). At 7 cm and 9 cm, moderate and severe fatty infiltration had lower Emeans and Emodes than normal liver (p < 0.001). The results suggest that digital analysis of hepatic ultrasonogram can be useful in the evaluation of the degree of fatty infiltration of the liver in dairy cattle.     

Comparison of estrus induction and subsequent fertility with two different intravaginal devices in ewes during the non-breeding season

Two experiments were conducted to compare the effect of estrus induction by controlled internal drug release (CIDR) and intravaginal cream containing 500 mg progesterone (P cream) in ewes during the non-breeding season. In the first experiment, twenty-four ewes were randomly grouped for two treatments with the different intravaginal devices for 12 days: Group A was the CIDR group and Group B was the P cream group. Blood was collected from all treated ewes, and progesterone (P(4)), estradiol 17-beta (E(2)) and luteinizing hormone (LH) concentrations were measured by enzyme immunoassay. In the second experiment, the conception rates from natural mating, estrus-detected AI (inseminated 12 h after estrus detection), or fixed-time AI (inseminated 42 h after removal of an intravaginal device) in 127 ewes treated with CIDR or P cream were compared. In Experiment 1, the rate of estrus induction and the time of estrus onset after device removal were 91.7% and 36.3 +/- 15.7 h in Group A, and 100% and 35.0 +/- 12.6 h in Group B, respectively. There were no significant differences between the devices. The mean plasma P(4) concentration in Group B was significantly (P < 0.01) lower than Group A between day -9 and day -1 (Day 0: the day of device removal). However, no significant differences were found in the mean E(2) concentrations of the two groups after treatment. The mean time of estrus onset in ewes with an observed LH surge and the time of LH surge after treatment were 23.3 +/- 8.7 h and 30.3 +/- 5.0 h for Group A and 27.6 +/- 6.5 and 26.3 +/- 8.0 h for Group B, respectively, and there were no significant differences. However, a significant difference (P < 0.05) was found in the mean time from the time of estrus onset to LH surge between Group A (6.4 +/- 6.7 h) and Group B (-1.3 +/- 4.1 h). In Experiment 2, the conception rates for natural mating, estrus-detected AI, and fixed-time AI were 55.0, 29.4, and 25.0% for Group A and 40.7, 25.0, and 42.1% for Group B, respectively, and there were no significant differences. These results suggest that the effect of induction of estrus and ovulation and the rate of conception after treatment were comparable to CIDR even though the plasma P(4) concentration of the P cream method tended to be low during the insertion period.     

Recovery of the everted cystacanth of seven acanthocephalan species of birds from feral raccoons (Procyon lotor) in Japan

Acanthocephalan everted cystacanths were detected in the intestines of 61 out of 555 feral raccoons (Procyon lotor) trapped between May 2003 and April 2005 in the western region of mainland Japan (Honshu). All collected specimens were identified as the species of birds including 3 Centrorhynchus species (C. bazaleticus, C. elongatus, and C. teres), Sphaerirostris lanceoides, Plagiorhynchus ogatai, Porrorchis oti, and Southwelina hispida. Three species, C. bazaleticus, C. teres, and S. lanceoides, were new to distribution records in Japan and the Far East. Recovery rates of these acanthocephalans were higher from the raccoons trapped in spring and early summer (range 8.3-36.8%, average 21.5%) than during the remainder of the year (range 0-20.0%, average 6.3%). The reason for this seasonal change is unknown. There is little information on the acanthocephalan fauna of wild birds in Japan, however, recovery of the cystacanths from the feral raccoons captured through the year might provide useful information on the fauna and geographical distribution of avian acanthocephalans.     

Epidemiological study on periodontal diseases and some other dental disorders in dogs

The prevalence of dental disorders in dogs was studied by applying index systems for human with some modifications. A total of 251 mongrel dogs including 143 stray dogs kept in the Animal Protection Offices in Tokyo and Hokkaido and 108 pet dogs visiting veterinary clinicians in Chiba Prefecture and Hokkaido were used. Periodontitis was prevalent among these dogs regardless of their sources and its incidence was increased with age. The lesion was more severe and more frequent in the premolar and molar regions than in the maxillary and mandibular incisor regions. Missing of teeth was observed at a high and increasing incidence with age. The tooth most commonly lost was the first premolar, followed by the other premolars and molars, where severe periodontitis was frequently found. Calculus was seen on many teeth, and aging agravated its prevalence and severity. Dental caries was observed in stray dogs, but neither to a serious degree nor at a significant level. These findings emphasize the necessity of dental hygiene, proper dental care and continuous periodical survey for dogs.     

Molecular study on chicken tumor necrosis factor receptor-II and tumor necrosis factor receptor-associated factor-5

Tumor necrosis factor (TNF) receptor-associated factors (TRAFs) were identified as signal transducers for the tumor necrosis factor receptor (TNFR) superfamily. In this study, we cloned and characterized two genes that encode chicken TNFR-II and TRAF5. The initial cDNA fragments were obtained by suppressive subtractive hybridization (SSH) of chicken spleen cells with or without lipopolysaccharide stimulation (Salmonella typhimurium SL1181 (RE-mutant)). The results showed that chicken TNFR-II is 1518 bp in length with an open reading frame (ORF) of 1386 bp having 31% homology with human TNFR-II. Expression analysis of chicken TNFR-II revealed that it is highly expressed in the spleen and bursa of Fabricius. The chicken cell lines IN24, MSB1 and 1104B express TNFR-II abundantly. The time course analysis of expression in spleen, bursa of Fabricius and IN24 cell line showed that TNFR-II is maximally expressed at 6 h after stimulation in bursa of Fabricius and after 8 h stimulation in the IN24 cell line. With regard to TRAF5, the complete sequence was 1936 bp in length with an ORF of 1671 bp that showed 71.3% homology with human TRAF5. Expression analysis showed that, among the tissues examined, TRAF5 was strongly expressed in spleen and bursa of Fabricius, while among the cell lines examined, it was maximally expressed in IN24. Thus, both genes were expressed in the same tissues and cell line among examined materials. These results suggest that chicken TNFR-II may interact with TRAF5 adaptor protein to complete its signal transduction pathway.     

Acquisition of quinolone resistance and point mutation of the gyrA gene in Campylobacter jejuni isolated from broilers and in vitro-induced resistant strains

A dramatic rise in the number of resistant Campylobacter to quinolones has been documented in human patients and domestic animals. In this study, the mechanism of acquisition of quinolone resistance was studied by detecting point mutations in the gyrA gene of Campylobacter strains obtained from broilers and strains with in vitro-induced resistance. The minimal inhibitory concentrations (MICs) of norfloxacin (NFLX) and ofloxacin (OFLX) for the strains that had no point mutation were slightly increased from the source strain (Campylobacter jejuni ATCC 33560). The MICs of nalidixic acid (NA), NFLX, and OFLX for the strains that had the point mutation at Thr-86 were 100 or 200 microg/ml, 50 microg/ml, and 25 microg/ml, respectively. The MIC of NA for the strain that had a point mutation at Asp-90 higher than those for the strains that had the point mutation at Thr-86, but the MICs of NFLX and OFLX were relatively lower than those for the strains that had point mutation at Thr-86. These findings suggest that the degree of antimicrobial resistance against NA, NFLX, and OFLX in the in vitro-induced C. jejuni strains was associated with the location of the point mutation in gyrA. On the other hand, a point mutation in all seven resistant strains isolated from broilers was located only at Thr-86, while the MICs of the three quinolones varied in each wild strain. This suggests that another mechanism might also be involved in the acquisition of quinolone resistance in C. jejuni wild strains.     

Chicken peripheral blood CD3+CD4-CD8- cells are regulated by endocrine and nerve systems

The existence of CD3(+)CD4(-)CD8(-) T cells in thymus and spleen has already been known. However, because of the presence of large amounts of thrombocytes in peripheral blood (PB), the proportion of CD3(+)CD4(-)CD8(-) T cells in PB has yet to be investigated. Therefore, the proportion of peripheral T cell-subsets was investigated in 6-week-old chickens. The percentage of CD3(+) cells, CD4(+) cells, CD8 alpha(+) cells, CD8 beta(+), and CD3(+)CD4(-)CD8(-) cells was 76%, 41%, 14%, 5%, and 15%, respectively. The proportion of CD3(+)CD4(-)CD8(-) cells in PB increased during egg-laying periods and in chickens treated with an analog of estrogen, while it decreased with age and in response to restraint stress. All of the CD3(+)CD4(-)CD8(-) cells expressed TCR1, and did not have NK activity. CD3(+)CD4(-)CD8(-) cells represent about 60% of peripheral TCR1(+) cells. These findings indicate that the proportion of CD3(+)CD4(-)CD8(-) cells is regulated by the endocrine and nerve systems.