Hepatic tyrosine aminotransferase activity is affected by chronic heat stress and dietary tyrosine in broiler chickens

1. Two experiments were conducted to investigate the impact of high temperature and dietary tyrosine (Tyr) content on performance and activity of hepatic tyrosine aminotransferase (EC 2.6.1.5.), an enzyme that catalyses the first step in the metabolic degradation of Tyr in broiler chickens. 2. Two-week-old birds were allocated to one of three temperature treatments: 24 degrees C (control), 36 degrees C (heat stress, HS) and 24 degrees C pair-fed (24PF) for 2 weeks and fed on diets containing 100% (Experiment 1) and 50, 100 and 200% (Experiment 2) of the NRC requirement for Tyr. 3. In Experiment 1, exposure of chickens to 36 degrees C for 2 weeks caused significant increase in hepatic tyrosine aminotransferase activity but no significant change in activity of hepatic phenylalanine 4-hydroxylase (EC 1.14.16.1) (an enzyme that catalyses conversion of phenylalanine to Tyr) compared with the 24PF birds. No significant changes attributable to heat stress were detected in hepatic glutamic-oxaloacetic transaminase (EC 2.6.1.1) activity. 4. In Experiment 2, heat stress caused reductions in weight gain and feed intake in chickens on all diets, compared with their control counterparts. Hepatic tyrosine aminotransferase activity was increased by heat stress compared with their 24PF counterparts in chickens fed on the 100 and 200% Tyr diets, while in chickens fed the 50% Tyr diet, it was reduced by heat stress. 5. From these results, it is suggested that hepatic tyrosine aminotransferase activity is affected by heat stress and dietary Tyr content and the increased tyrosine aminotransferase activity with, in part, relatively low phenylalanine hydroxylase activity in hepatic tissues may be involved in the Tyr metabolism characteristic of heat-stressed chickens.     

Isolation and serological survey of Salmonella in pigs in Japan

A total of 267 fecal and serum samples collected from individual pigs reared on a Salmonella-positive farm were subjected to bacteriological and serological examinations of Salmonella. Salmonella was isolated from 47 pigs (17.6%) and prevalence of antibody to lipopolysaccharide (LPS) of S. Typhimurium, which was partly common to S. O4, 12: d: -, was observed in 90 pigs (33.7%). Salmonella was isolated from 26 (28.9%) of 90 antibody-positive pigs and 21 (11.9%) of 177 antibody-negative pigs. Twenty-one of 36 pigs (58.3%) positive for S. O4, 12: d: -, five of 10 pigs (50.0%) positive for S. Havana, and none for S. Anatum had antibodies. Thus, seropositive rates were higher than isolation-positive rates, and antibody prevalence was associated with serovars of the isolates. Then, we analyzed antibody prevalence among pigs on Japanese pig farms. The antibodies to LPS of S. Typhimurium were found in 195 of 1,498 pigs (13.0%) and in at least one serum sample on 35 of 52 farms (67.3%). Our results indicate that Salmonella does not seem to be so prevalent in pigs though it is widely prevalent among pig farms.     

Serum activities of tartrate-resistant acid phosphatase and bone specific alkaline phosphatase as indices of bone metabolism in the cow

The correlation between the serum hydroxyproline concentration and serum activity levels of TRAP and BALP was examined in 41 cows. The correlated coefficient (r) was 0.6391 for TRAP and 0.3147 for BALP, respectively. Judging from the significant correlation to the serum hydroxyproline concentration, serum TRAP activity is an index for bone metabolism in cows. Serum TRAP activity was therefore measured in 205 healthy cows (2-9 years old) in order to observe the changes in bone resorption with aging and milk production. TRAP levels differed slightly between group A (< or =4 yrs) and B (5 yrs< or =) at the same stage of lactation. The activity levels rose slightly at the height of lactation stage and during the dry stage.     

Parent‐of‐origin effects on carcass traits in Japanese Black cattle

Variances caused by the differential expression of paternally and maternally imprinted genes controlling carcass traits in Japanese Black cattle were estimated in this study. Data on marbling score (BMS), carcass weight, rib thickness, rib‐eye area (REA) and subcutaneous fat thickness (SFT) were collected from a total of 13,115 feedlot steers and heifers in a commercial population. A sire–maternal grandsire model was used to analyse the data, and then, imprinting parameters were derived by replacing the genetic effect of the dam with the effect of the maternal grandsire in the imprinting model to calculate the genetic parameter estimates. The proportions of the total genetic variance attributable to imprinted genes ranged from 8.7% (SFT) to 35.2% (BMS). The remarkably large imprinting variance of BMS was mainly contributed by maternally expressed inheritance because the maternal contribution of the trait was much larger than that of the paternal trait. The parent‐of‐origin effect originating from maternal gene expression was also observed for REA. The results suggested the existence of genomic imprinting effects on the traits of the Japanese Black cattle. Hence, the parent‐of‐origin effect should be considered for the genetic evaluation of Japanese Black cattle in breeding programmes.     

Distribution of immunoglobulin-containing cells in calves inoculated orally with ruminal Bacteroides succinogenes and Selenomonas ruminantium

Distribution of immunoglobulin(Ig)-containing cells was investigated in calves inoculated orally with live organisms of both Bacteroides succinogenes and Selenomonas ruminantium. Pathological changes and many Ig-containing cells were observed in calves which inoculated three times at 2, 3 and 26 days of age. Follicular germinal center was increased in number and size of the lymph nodes associated with the forestomach, suggesting activation of lymph apparatus. In the associated lymph nodes, IgG-containing cells were predominant and were located in both cortex and medulla, mainly in the medullary cord, B lymphocyte areas. Only a few IgA- and IgM-containing cells were observed in the lymph nodes. Accordingly, the inoculated bacteria may stimulate IgG-containing B lymphocyte populations. A few IgG-containing cells were detected in the mucosa of the forestomach. Ig-containing cells, predominantly IgG, were observed in the mucosa of the abomasum and intestine, and in the mesenteric lymph nodes. However, number of the cells in the mesenteric lymph nodes was smaller than that of the forestomach associated lymph nodes. The results suggest that the intraorally inoculated bacteria may stimulate the maturation of IgG positive lymphocytes in the lymph nodes associated with the forestomach.     

Spindle formation and microtubule organization during first division in reconstructed rat embryos produced by somatic cell nuclear transfer

The present study was conducted to demonstrate the spindle formation and behavior of chromosomes and microtubules during first division in reconstructed rat embryos produced by somatic cell nuclear transfer (SCNT) with cumulus cell nuclei. To demonstrate the effect of oocyte aging after ovulation on the cleavage of SCNT embryos, micromanipulation was carried out 11, 15 and 18 h after injection of hCG. SCNT oocytes were activated by incubation in culture medium supplemented with 5 microM ionomycin for 5 min followed by treatment with 2 mM 6-dimethylaminopurine (6-DMAP) in mR1ECM for 2-3 h. For immunocytochemical observation, the SCNT embryos were incubated with monoclonal anti-alpha-tubulin antibody and then fluorescein isothiocyanate (FITC)-conjugated goat anti-mouse IgG. Cleavage rates were significantly higher for oocytes collected after 15 and 18 h rather than for those collected 11 h after injection of hCG (56 and 53%, respectively vs. 28%; P<0.05). Premature chromosome condensation occurred before activation of the SCNT oocytes, but adequate spindle formation was only rarely observed. The distribution of microtubules in SCNT embryos after activation was different from those of fertilized and parthenogenic oocytes, i.e., a dense microtubule organization shaped like a ring was observed. Eighteen to 20 h post-activation, most SCNT embryos were in the 2-cell stage, but no nucleoli were clearly visible, which was quite different from the fertilized oocytes. In addition, first division with and without small cellular bodies containing DNA was observed in the rat SCNT embryos in some cases. The present study suggests that reorganization of transferred nuclei in rat SCNT embryos may be inadequate in terms of formation of the mitotic assembly and nucleolar reorganization.     

Effect of modification of ovary preservation solution by adding glucose on the maturation and development of pig oocytes after prolonged storage

Oocytes lose their developmental competence during prolonged storage of the ovary. In the present study, we supplemented the preservation solution for pig ovaries (phosphate buffered saline, PBS) with glucose and preserved the ovaries for 6 h at 25 C. Subsequently, we examined the glucose concentration of the follicular fluid (FF), pH of the FF, survival rate of the granulosa cells, and maturation and developmental competence of oocytes after storage. During storage, the glucose concentration of the FF (2.1 mM), pH of the FF (7.4), and survival rate of the granulosa cells (69.5%) rapidly decreased (glucose concentration: under 1.1 mM; pH: 6.8; and survival rate: 43%). On the other hand, when the preservation solution was supplemented with glucose (15 mM), the glucose concentration of the FF increased and the survival rate of the granulosa cells improved, although the pH of the FF decreased further (from 6.8 to 6.6). In addition, supplementation with glucose significantly improved the rates of oocytes at metaphase II (0 h: 65.0%; 6 h without glucose: 23.8%; and 6 h with glucose: 43.8%) and attenuated the decline in the rates of fertilization and development that resulted from prolonged storage, although there were no significant differences. In conclusion, modification of the preservation solution by the addition of glucose increased the glucose concentration of the FF and improved the rate of maturation of pig oocytes.     

Comparison of two different nutritive conditions against the changes in peripheral blood mononuclear cells of periparturient dairy cows

To clarify the relationship between cellular immune status and nutritive condition in periparturient dairy cows, feeding content, blood profiles, and immune condition were observed in cows from two dairy herds with different types of feed content. Immunological analyses such as leukocyte population and peripheral blood mononuclear cell (PBMC) mRNA of IFN-gamma, TNF-alpha, IL-4, and IL-10, quantified by real-time RT-PCR were performed. With regard to feed content during dry periods, there were six cows in the herd with insufficient non-structural carbohydrate (NFC) intake (group I) and six cows in the herd with sufficient NFC intake (group II). Significantly lower levels of blood glucose were observed in group I between weeks -12 and 16 compared with group II. Serum cholesterol level was significantly lower in group I between weeks 2 and 10 than in group II. The numbers of CD3+ and CD4+ T cells in group I were significantly lower than those in group II in weeks 6 and 14. The numbers of CD21+ B cells were significantly lower in group I than in group II in weeks -16, -12, 2, and 10. On the other hand, the CD4+/CD8+ ratio in group II was significantly higher than group I between weeks 2 and 14. The IFNgamma/IL-4 mRNA rate in group I was significantly lower than group II in week 6. We concluded that cellular immune depression occurrs after calving in dairy cows with low nutritional status in the periparturient period.     

Matrix metalloproteinases-2 and -9 activities in bovine follicular fluid of different-sized follicles: relationship to intra-follicular inhibin and steroid concentrations

Matrix metalloproteinases (MMPs) play very important roles in extracellular matrix (ECM) remodeling during ovarian follicular development, ovulation and atresia. The aim of the present study was to determine the content of gelatinases in follicular fluid in various sized bovine follicles. Bovine ovaries were collected from local slaughterhouse and follicular fluid from follicles of 2 to over 25 mm in diameter was collected. Gelatinase activity within the follicular fluid was analyzed by gelatin zymography. The concentration of inhibin in the follicular fluid was also measured by immunoblot analysis. The proMMP-2 and alpha-subunit (alphaN) inhibin was detected in all follicles regardless of their size. The abundance of proMMP-2 varied with follicular size, while alphaN inhibin increased significantly (P<0.01) in follicles of 10-14 and 15-20 mm in size. There was a positive and negative correlation between estradiol (E(2)) and progesterone (P(4)) concentrations with abundance of proMMP-2, respectively. Follicles of diameter over 25 mm had greater proMMP-9 activity than other follicles. These same follicles had significantly (P<0.01) lower inhibin levels than follicles of 10-14 and 15-20 mm in size. In conclusion, these results suggest a significant role of these proteases in growth and development of bovine follicle, particularly proMMP-2 and active MMP-2 activities in the follicular fluid could serve as markers of follicular health while abundance of proMMP-9 may possibly denote a follicular cyst.     

Effect of dietary addition of seaweed and licorice on the immune performance of pigs

In pig production, dietary additive antibiotics are usually used for growth stimulation and disease prevention, although there is public concern about the increased incidence of resistant antibiotics and food safety. It is possible that such antibiotics might be replaced by naturally derived products such as seaweed and licorice. In this study, we evaluated the effect of dietary addition of seaweed and licorice on enhancing the immune function in swine. The animals of each group (eight animals per group) were sensitized at day 42 and 49, and the immunoglobulin production and the expression of cytokines were detected by the ELISA and real‐time PCR. As the results, saliva IgA production of the seaweed‐treated group increased around five times compared to that of control (day 56). Delayed hypersensitivity reaction and IgG production of the seaweed‐treated group increased around 1.8–2.0 times. In addition, enhanced saliva IgA production was detected at day 50 (around two times) and day 51 (around five times) by the licorice treatment, and lower expression level of tumor necrosis factor‐α messenger RNA at day 51 (around 1/25) was observed in the licorice treatment. We conclude that the replacement of antibiotics by naturally derived dietary additives might be feasible for immune system enhancement.