Congruency of phylogenies derived from different proteins

Summary This communication examines the question of phylogenetic congruency- i.e., whether or not the branching order of evolutionary trees is independent of the protein studied. It was found that trees constructed for birds on the basis of immunological comparison of their transferrins, albumins, and ovalbumins agree approximately with a published tree based on the amino acid sequences of their lysozymesc. This congruency is especially noteworthy with respect to the phylogenetic position of the chachalaca, a Mexican bird classified on morphological grounds in the family Cracidae of the order Galliformes. At the protein level, this species differs as much from non-cracid galliform birds as does the duck, which belongs to another order. Despite the organismal similarity between cracid and non-cracid galliform birds, the molecular relationship is remote. If this contrast between organismal and molecular results had been based on comparative studies with only lysozyme, one could have ascribed the contrast to the possibility that chachalaca lysozyme was paralogous, rather than orthologous, to the other bird lysozymesc. Examination of several proteins is thus desirable in cases of possible paralogy.This work was supported in part by grants GB-42028X from NSF and GM-21509 from NIH     

Plasmodium gallinaceum: response of malarious chicken embryos to injection of serum from hyperimmunized chickens.

The injection of serum from chickens hyperimmune to the avian malaria parasite, Plasmodium gallinaceum, or of nonimmune serum into the chorioallantoic cavity of chicken embryos infected with the same parasite resulted in changes in the blood picture of such embryos. In those embryos receiving serum from hyperimmunized birds: The level and rate of parasite increase were depressed; the maturation of the erythroid elements was reduced; the hematocrit values increased only by 27% as compared with 69% in untreated embryos. While hematocrit levels and rates of erythrocytic maturation were depressed in those embryos injected with nonimmune serum, there was no suppression of parasitemia. The results suggest a definite role of immunity in the anemia accompanying malaria.     

细胞色素P450 3A7羟化维生素D3的功能研究

本研究通过体外生化实验研究细胞色素P450 3A7对维生素D3的羟化作用。根据GenBank报道的序列设计特异引物,扩增cyp3a7的编码区,将cyp3a7的编码区插入到pcDNATM3.1/myc-His(-) A的XhoⅠ/Bam HⅠ,通过测序检测序列的正确性。pcDNA-CYP3A7及pcDNA分别瞬时转染293T细胞,48 h后收集细胞,提取S9组分,用Bradford法测定蛋白质浓度。S9组分经12%SDS-PAGE凝胶电泳和Western blotting检测,用myc抗体作为一抗检测CYP3A7在293T细胞的表达水平。0.6 mg S9组分与1μmol/L维生素D3于37℃孵育30 min,用4倍体积的氯仿甲醇(体积比为3∶1)抽提,有机相在氮气流下吹干,残基用于HPLC分析。结果显示,重组表达CYP3A7的293T细胞的S9组分通过Western blotting检测到了特异的约60 kD的条带,对照样品未检测到特异条带的蛋白质。重组表达CYP3A7的293T细胞S9组分的孵育样品通过HPLC检测到了25-羟基维生素D3,对照样品未检测到25-羟基维生素D3。结果表明重组表达的CYP3A7羟化维生素D3生成25-羟基维生素D3。本研究为进一步探究还有哪些P450参与维生素D3在鸡体内的代谢,为阐明其代谢途径提供理论依据。     

The effects of supplemental levels of inorganic zinc,manganese and copper on growth performance in commercial broilers

Commercial broiler premixes provide trace minerals (TMs) such as Zinc (Zn), Manganese (Mn) and Copper (Cu) above the birds' requirements to maximise broiler performance. High inclusion levels of TM and their low absorption in the broiler's gastrointestinal tract have led to increased TM levels in their litter, raising concerns about TM's accumulation in the environment from poultry litter. A 35-day broiler trial was conducted at a broiler facility with 2 880-day-old Cobb 500 broiler males. The objective was to evaluate whether decreasing inorganic Zn, Mn and Cu levels in broiler diets would have a negative effect on broiler growth parameters such as BW, average daily gain, cumulative feed intake (FI) and feed conversion ratio (FCR). Consequently, these minerals were supplemented at 100%, 50%, 25%, and 0% of the Cobb standards. No significant difference in FI and FCR was observed between the treatments for the first 21 days, and there were no significant differences in BW between the positive control, negative control (NC), and 50% of Cobb recommended levels at day 35. However, BW for the NC was significantly higher than the 25% level. These results show that reducing TM levels in broiler diets below commercially recommended levels had no negative effect on broiler performance, with the subsequent economic benefits for producers.     

A Single Nucleotide Polymorphism of Chicken Acetyl-CoA Carboxylase A Gene Associated with Fatness Traits

Acetyl-CoA carboxylase α (ACCα) is a major rate-limiting enzyme in the biogenesis of long-chain fatty acids. It can catalyze the carboxylation of acetyl-CoA to form malonyl-CoA that plays a key role in the regulation of fatty acid metabolism. The objective of the present study was to investigate the associations of ACCα gene polymorphisms with chicken growth and body composition traits. The Northeast Agricultural University broiler lines divergently selected for abdominal fat content and the Northeast Agricultural University F₂ Resource Population were used in the current study. Body weight and body composition traits were measured in the aforementioned two populations. A synonymous mutation was detected in the exon 19 region of ACCα gene, then polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) method was developed to genotype all the individuals derived from the aforementioned populations. Association analysis revealed that the polymorphism was associated with abdominal fat weight and percentage of abdominal fat in the two populations. The results suggested that ACCα gene could be a candidate locus or linked to a major gene that affects abdominal fat content in the chicken.     

ISOLATION OF 105 MICROSATELLITE LOCI FROM AN OVINE GENOMIC LIBRARY ENRICHED FOR MICROSATELLITES

This paper reports on the development of a small-insert (～700 bp) total-genomic library for sheep specifically designed for enrichment for microsatellite (ms) loci. Four enriched libraries were prepared by amplification of the primary library with CA₁₅, CA₁₁, TG₁₅ and TG₁₁ oligonucleotide primers. A total of 11,020 clones was recovered, screened for dinucleotide repeats and over 500 positive clones sequenced. Sequence analysis indicated low clone redundancy and yielded 105 new ovine ms loci. Seventy-two percent of the new loci were found to be polymorphic in the sires of the AgResearch International Mapping Flock (IMF). The 105 new microsatellite loci increase the number of microsatellites available for sheep by >7%.     

Purification and Separation of A and B Forms of N-Acetyl-β-D-Hexosaminidase from Human Aorta

Human aorta has been shown to possess multiple forms of N-Acetyl-6-D-hexosaminidase (β-2-acetamido-2-deoxy-D-glucoside-acetamido-deoxyglucohydro-lase, EC 3.2, 1.30). The enzyme was separable, by gel electrophoresis, into 2 enzymatically active bands representing A and B forms. By gel electro-focussing, A and B forms were further subdivided into at least 5 and 8 bands, respectively. The B form consisted of 4 bands (B₁) and 4 bands (B₂), which were not inactivated at 50° for 3 hr. (at pH 4.4) in the presence of serum; whereas, the 5 bands found in A form were completely inactivated. All forms of the enzyme were active towards naphthol-AS-BI-N-acetyl-β-D-glucosaminide and the corresponding galactosaminide (about one-eighth of the hydrolysis rate of the former), suggesting each single enzyme acts on both substrates. The N-acetyl-hexosaminidases of bull epididymis, by comparison, were also found to be active towards both substrates and to possess 13 bands having pis more alkaline than those of the B form of the human enzyme, By heat inactivation we found that the aortic enzyme consisted of 51% of A and 49% of B (B₁ + B₂ .). Neuraminidase had no effect on either form of the aortic preparation. Both forms were partially purified and separated by conventional methods. They required BSA for their maximal activity; the A form being more dependent BSA than the B form, With PNP-N-acetyl-β-D-glucosaminide and the corresponding galactosaminide, Km of 1.04 mH and 0.54 mM, respectively, for A form and of 1.74 and 1.48 mM, respectively, for B form were obtained. While the purified B form was stable and did not transform into other species, the purified A form gradually transformed into B form as well as into other new forms during storage at -20°.     

Sine Wave Extremely Low Frequency Magnetic Fields Protect Chick Embryos Against UV-Induced Death

Four-day-old chicken embryos were exposed to extremely low frequency (ELF) magnetic fields (MF) prior to UV exposure (75 min, predominantly UV-C, 0.4 mW/cm²) to investigate possible MF-mediated protection against lethal effects of UV. The UV exposure typically resulted in a 20% survival rate (as judged by beating hearts) in sham-exposed embryos 3 h postexposure. In contrast, exposure to a 50 (10, 50, or 100 µT) or 60 Hz (10 µT) vertical MF caused a significant increase in survival rate, observed only 30 min after UV exposure. No difference in protection levels was seen between these exposure intensities. A horizontal 50 Hz MF (10, 50, or 100 µT) did not result in the general protection against UV-induced death observed for vertical fields, suggesting that the size of the induced electric field (which differs between horizontal and vertical exposure) is important for the MF-induced protection. To explore the molecular mechanisms involved in this effect, immunoblotting experiments with an antibody against the inducible form of hsp70 were performed. These showed that application of MF (50 Hz, 200 µT, 1 h) induced hsp70 expression in human K562 cells.     

Domestication‐related variation in social preferences in chickens is affected by genotype on a growth QTL

A growth‐related QTL on chicken chromosome 1 has previously been shown to influence domestication behaviour in chickens. In this study, we used Red Junglefowl (RJF) and White Leghorn (WL) as well as the intercross between them to investigate whether stress affects the way birds allocate their time between familiar and unfamiliar conspecifics in a social preference test (‘social support seeking’), and how this is related to genotype at specific loci within the growth QTL. Red Junglefowl males spent more time with unfamiliar chickens before the stressful event compared to the other birds, whereas all birds except WL males tended to spend less time with unfamiliar ones after stress. A significant QTL locus was found to influence both social preference under undisturbed circumstances and social support seeking. The WL allele at this QTL was associated not only with a preference for unfamiliar individuals but also with a shift towards familiar ones in response to stress (social support seeking). A second, suggestive QTL also affected social support seeking, but in the opposite direction; the WL allele was associated with increased time spent with unfamiliar individuals. The region contains several possible candidate genes, and gene expression analysis of a number of them showed differential expression between RJF and WL of AVPR2 (receptor for vasotocin), and possibly AVPR1a (another vasotocin receptor) and NRCAM (involved in neural development) in the lower frontal lobes of the brains of RJF and WL animals. These three genes continue to be interesting candidates for the observed behavioural effects .