Interaction of proteinase inhibitors with Cry1Ac toxicity and the presence of 15 chymotrypsin cDNAs in the midgut of the tobacco budworm, Heliothis virescens (F.) (Lepidoptera: Noctuidae)

BACKGROUND: The potential development of resistance to Bacillus thuringiensis (Bt) cotton and surging of non‐targeted insects is a major risk in the durability of Bt plant technology. Midgut proteinases are involved in Bt activation and degradation. Proteinase inhibitors may be used to control a wide range of insects and delay Bt resistance development. Proactive action to examine proteinase inhibitors for synergistic interaction with Bt toxin and cloning of proteinase cDNAs for RNAi is necessary to make transgenic cotton more versatile and durable. RESULTS: A sublethal dose (15 ppb) of Cry1Ac, 0.5% benzamidine and 0.02% phenylmethylsulfonyl fluoride significantly suppressed midgut azocaseinase, tryptic and chymotryptic activities, and resulted in reductions in larval and pupal length and mass of Heliothis virescens. The combination of proteinase inhibitor and Bt suppressed 20–37% more larval body mass and 26–80% more enzymatic activities than the inhibitor only or Bt only. To facilitate knockdown‐resistance‐related proteinase genes, 15 midgut chymotrypsin cDNAs were sequenced. Most predicted chymotrypsins contained the conserved N‐termini IVGG, three catalytic center residues (His, Asp and Ser), substrate specificity determinant (Ser or Gly) and cysteines for disulfide bridges. These putative chymotrypsins were separated into three distinct groups, indicating the diverse proteinases evolved in this polyphagous insect. CONCLUSION: H. virescens has evolved diverse midgut proteinase genes. Proteinase inhibitors have potential insecticidal activity, and the interaction of Bt with proteinase inhibitors is desirable for enhancing Bt toxicity and delaying resistance development. Intensive sequencing of chymotrypsin cDNAs will facilitate future functional examinations of individual roles in Bt toxicity and resistance development and facilitate targeted control using RNAi and/or proteinase inhibitors. Copyright © 2012 Society of Chemical Industry     

Kaempferol improves carcase characteristics in broiler chickens by regulating ANGPTL3 gene expression

1. The objective of this study was to examine the beneficial effects of kaempferol, a naturally occurring polyphenol, on carcase characteristics in broiler chickens and the mechanisms involved in this regulation.

2. Broiler chickens were randomly divided into 7 groups: control, carrier control, kaempferol (0·3%), kaempferol (0·6%), hypercholesterolemic (HLD), HLD and kaempferol (0·3%), HLD and kaempferol (0·6%).

3. Seven weeks after treatment, carcase characteristics, lipid levels in the blood and liver, expression of hepatic Angiopoietin-like protein 3 (ANGPTL3) mRNA, and expression of adipose lipoprotein lipase (LPL) protein were determined.

4. Treatment with kaempferol (0·3 or 0·6%) significantly increased plasma high-density lipoprotein cholesterol levels, decreased percentage of abdominal fat, thickness of subcutaneous fat, plasma and hepatic total cholesterol and triglyceride concentrations, muscle malondialdehyde level and down-regulated expression of ANGPTL3 mRNA concomitantly with up-regulated expression of LPL protein in normal and hypercholesterolemic broiler chickens.

5. Kaempferol (0·3 or 0·6%) treatment had no significant effect on the values of percentage of breast muscle, percentage of leg muscle, carcase weight and eviscerated percentage.

6. The results suggest that kaempferol improves carcase characteristics by decreasing expression of ANGPTL3 in broiler chickens.     

Modeling the relationships between quality and biochemical composition of fatty liver in mule ducks

The fatty liver of mule ducks (i.e., French "foie gras") is the most valuable product in duck production systems. Its quality is measured by the technological yield, which is the opposite of the fat loss during cooking. The purpose of this study was to determine whether biochemical measures of fatty liver could be used to accurately predict the technological yield (TY). Ninety-one male mule ducks were bred, overfed, and slaughtered under commercial conditions. Fatty liver weight (FLW) and biochemical variables, such as DM, lipid (LIP), and protein content (PROT), were collected. To evaluate evidence for nonlinear fat loss during cooking, we compared regression models describing linear and nonlinear relations between biochemical measures and TY. We detected significantly greater (P = 0.02) linear relation between DM and TY. Our results indicate that LIP and PROT follow a different pattern (linear) than DM and showed that LIP and PROT are nonexclusive contributing factors to TY. Other components, such as carbohydrates, other than those measured in this study, could contribute to DM. Stepwise regression for TY was performed. The traditional model with FLW was tested. The results showed that the weight of the liver is of limited value in the determination of fat loss during cooking (R(2) = 0.14). The most accurate TY prediction equation included DM (in linear and quadratic terms), FLW, and PROT (R(2) = 0.43). Biochemical measures in the fatty liver were more accurate predictors of TY than FLW. The model is useful in commercial conditions because DM, PROT, and FLW are noninvasive measures.     

Growth hormone stimulates protein synthesis in bovine skeletal muscle cells without altering insulin-like growth factor-I mRNA expression

Growth hormone is a major stimulator of skeletal muscle growth in animals, including cattle. In this study, we determined whether GH stimulates skeletal muscle growth in cattle by direct stimulation of proliferation or fusion of myoblasts, by direct stimulation of protein synthesis, or by direct inhibition of protein degradation in myotubes. We also determined whether these direct effects of GH are mediated by IGF-I produced by myoblasts or myotubes. Satellite cells were isolated from cattle skeletal muscle and were allowed to proliferate as myoblasts or induced to fuse into myotubes in culture. Growth hormone at 10 and 100 ng/mL increased protein synthesis in myotubes (P < 0.05), but had no effect on protein degradation in myotubes or proliferation of myoblasts (P > 0.05). Insulin-like growth factor-I at 50 and 500 ng/mL stimulated protein synthesis (P < 0.01), and this effect of IGF-I was much greater than that of GH (P < 0.05). Besides stimulating protein synthesis, IGF-I at 50 and 500 ng/mL also inhibited protein degradation in myotubes (P < 0.01), and IGF-I at 500 ng/mL stimulated proliferation of myoblasts (P < 0.05). Neither GH nor IGF-I had effects on fusion of myoblasts into myotubes (P > 0.1). These data indicate that GH and IGF-I have largely different direct effects on bovine muscle cells. Growth hormone at 10 and 100 ng/mL had no effect on IGF-I mRNA expression in either myoblasts or myotubes (P > 0.1). This lack of effect was not because the cultured myoblasts or myotubes were not responsive to GH; GH receptor mRNA was detectable in them and the expression of the cytokine-inducible SH2-containing protein (CISH) gene, a well-established GH target gene, was increased by GH in bovine myoblasts (P < 0.05). Overall, the data suggest that GH stimulates skeletal muscle growth in cattle in part through stimulation of protein synthesis in the muscle and that this stimulation is not mediated through increased IGF-I mRNA expression in the muscle.     

Evaluation of 1-stage and 2-stage selection in yellow perch I: genetic and phenotypic parameters for body weight of F1 fish reared in ponds using microsatellite parentage assignment

Two selection methods, 1-stage selection (OSS) and 2-stage selection (TSS), for improving efficiency and profitability of selective breeding of yellow perch were evaluated, through examining the genetic and phenotypic parameters for BW of F(1) fish using microsatellite parentage assignment in this study. Approximately 94% of the sampled yellow perch progeny were assigned to single parental pairs using 8 microsatellite markers, which confirmed the applicability of the communal rearing technique in yellow perch breeding. Within OSS, the genetic correlation between 1-yr-BW and 2-yr-BW was high (0.98), indicating that the growth of yellow perch recorded at yr 1 could predict their growth for yr 2. Also mean family BW and family EBV for BW between yr 1 and 2 were found to be significantly correlated, suggesting yr 1 fast-growing yellow perch families continued to be the fast growing families in yr 2. Two-year random fish undergoing TSS were significantly heavier (P < 0.01) than those undergoing OSS. In addition, top males and females with TSS were heavier (P < 0.01) than those with OSS. Based on these results we concluded that the TSS was more desirable and effective for yellow perch breeding compared with OSS in terms of improving selection efficiency and reducing costs.     

Cross‐Sectional Study of the Prevalence of Radiographic Degenerative Joint Disease in Domesticated Cats

Objective: To determine the prevalence of radiographic signs of degenerative joint disease (DJD) in a randomly selected sample of domestic cats. Study Design: Prospective observational study. Animals: Client‐owned cats. Methods: Cats (n=100) from a single practice and equally distributed across 4 age groups (0–5; 5–10; 10–15, and 15–20 years old) were randomly selected (regardless of heath status) and sedated for orthogonal radiographic projections of all joints and the spine. Quasi‐Poisson regression analysis was used to investigate the relationship between patient demographics, blood biochemistry, hematologic and urine analysis variables, and DJD severity. Results: Most (92%) cats had radiographic evidence of DJD; 91% had at least 1 site of appendicular DJD and 55% had ≥1 site of axial column DJD. Affected joints in descending order of frequency were hip, stifle, tarsus, and elbow. The thoracic segment of the spine was more frequently affected than the lumbosacral segment. Although many variables were significantly associated with DJD, when variables were combined, only the association between age and DJD was significant (P<.0001). For each 1‐year increase in cat age, the expected total DJD score increases by an estimated 13.6% (95% confidence interval: 10.6%, 16.8%). Conclusion: Radiographically visible DJD is very common in domesticated cats, even in young animals and is strongly associated with age. Clinical Relevance: DJD is a common disease of domesticated cats that requires further investigation of its associated clinical signs.     

Improving in sacco incubation technique to evaluate fresh forage for selecting fast‐degrading perennial ryegrass (Lolium perenne L.)

Grasses with fast fibre degradation are required by intensive pasture‐based animal production systems to maximize intakes and productivity. To select fast‐degrading elite cultivars, a repeatable, rapid and inexpensive screening method should be developed, so large numbers of samples can be evaluated. This study refined the experimental procedure for the in sacco incubation technique using fresh (not dried) perennial ryegrass (Lolium perenne L.). Pre‐ruminal incubation treatment and ratio of forage weight to the surface area of the in sacco bag have been tested to evaluate their effects on ryegrass degradation kinetic parameters in the bovine rumen. The timing of sampling and the number of sampling time points were also examined. Results indicated that warming the bags in water prior to incubation led to a faster dry‐matter (DM) degradation in the first 12 h. If ratio of forage to bag surface area was between 26 and 45 mg DM cm^?2, degradation parameters were not affected by bag fill. Sampling between 9 and 12 h was critical for determining degradation rate. From these results, an improved in sacco incubation procedure is recommended for screening of ryegrasses used for cultivar selection. The principles demonstrated here for ryegrass may be applicable to other forages, although the critical sampling times for measuring degradation rate are likely to differ.