Alternative methods for disposal of spent laying hens: Evaluation of the efficacy of grinding,mechanical deboning,and of keratinase in the rendering process

Besides the challenges of mortality and litter disposal, the poultry industry must find economical means of disposing of laying hens that have outlived their productive lives. Because spent hens have low market value and disposing of them by composting and burial is often infeasible, finding alternative disposal methods that are environmentally secure is prudent. The feasibility of grinding or mechanically deboning spent hens with and without prior mechanical picking was evaluated for the production of various proteinaceous by-product meals. The end products were analyzed for nutrient content and found to be high in protein (35.3–91.9% CP) and, with the exception of the feathers, high in fat (24.1–58.3%), making them potentially valuable protein and energy sources. After considering physical and economic feasibility, mechanical deboning was determined to be a logical first step for the conversion of spent hens into value-added by-product meals. Because the hard tissue fraction (primarily feathers, bones, and connective tissue) generated by mechanically deboning the hens presents the greatest challenge to their utilization as feedstuffs, attention was focused on technologies that could potentially improve the nutritional value of the hard tissue for use as a ruminant protein source. Traditional hydrolysis of this hard tissue fraction improved its pepsin digestibility from 74% to 85%; however, subsequent keratinase enzyme treatment for 1 h, 2 h, 4 h, or 20 h after steam hydrolysis failed to improve the pepsin or amino acid digestibility any further (P > 0.10). Enzyme hydrolysis did, however, increase the quantities of the more soluble protein fractions (A: 45.5, 46.6, 52.8, 51.6, and 55.8% of CP; B₁: 3.2, 9.8, 6.0, 4.6, and 4.1% of CP; B₂: 11.7, 18.1, 22.8, 29.6, and 22.0% of CP for 0, 1 h, 2 h, 4 h, and 20 h, respectively) and reduced quantities of the less soluble fractions (B₃: 30.2, 18.1, 10.8, 5.5, and 10.2% of CP; C: 9.4, 7.5, 7.6, 8.8, and 7.9% of CP for 0, 1 h, 2 h, 4 h, and 20 h, respectively). The protein digestibility of the steam hydrolyzed hard tissue fraction from the mechanical deboning of spent hens was found to be comparable to the digestibility of feather meal, but post-hydrolysis keratinase treatment did not improve feeding value for ruminants.     

Normal development of the tomato clownfish Amphiprion frenatus: live imaging and in situ hybridization analyses of mesodermal and neurectodermal development

The normal embryonic development of the tomato clownfish Amphiprion frenatus was analysed using live imaging and by in situ hybridization for detection of mesodermal and neurectodermal development. Both morphology of live embryos and tissue‐specific staining revealed significant differences in the gross developmental programme of A. frenatus compared with better‐known teleost fish models, in particular, initiation of somitogenesis before complete epiboly, initiation of narrowing of the neurectoderm (neurulation) before somitogenesis, relatively early pigmentation of melanophores at the 10–15 somite stage and a distinctive pattern of melanophore distribution. These results suggest evolutionary adaptability of the teleost developmental programme. The ease of obtaining eggs, in vitro culture of the embryo, in situ staining analyses and these reported characteristics make A. frenatus a potentially important model marine fish species for studying embryonic development, physiology, ecology and evolution.     

Electrochemical reduction of ferrous alpha-verdoheme in complex with heme oxygenase-1

The heme oxygenase (HO) reaction consists of three successive oxygenation reactions, i.e. heme to alpha-hydroxyheme, alpha-hydroxyheme to verdoheme, and verdoheme to biliverdin-iron chelate. Of these, the least understood step is the conversion of verdoheme to biliverdin-iron chelate. For the cleavage of the oxaporphyrin ring of ferrous verdoheme, involvement of a verdoheme pi-neutral radical has been proposed. To probe this hypothetical mechanism in the HO reaction, we performed electrochemical reduction of ferrous verdoheme complexed with rat HO-1 under anaerobic conditions. On the basis of the electrochemical spectral changes, the midpoint potential for the one-electron reduction of the oxaporphyrin ring of ferrous verdoheme was found to be -0.47+/-0.01 V vs the normal hydrogen electrode (NHE). Because this potential is far lower than those of both flavins of NADPH-cytochrome P450 reductase, and of NADPH, it is concluded that the one-electron reduction of the oxaporphyrin ring of ferrous verdoheme is unlikely to occur and that the formation of the pi-neutral radical cannot be the initial step in the degradation of verdoheme by HO. Rather, it appears more reasonable to consider an alternative mechanism in which binding of O(2) to the ferrous iron of verdoheme is the first step in the degradation of verdoheme.     

Molecular Pathogenesis of Wilson Disease Among Indians: A Perspective on Mutation Spectrum in ATP7B gene, Prevalent Defects, Clinical Heterogeneity and Implication Towards Diagnosis

Aims We aim to identify the molecular defects in the ATP7B, the causal gene for Wilson disease (WD), in eastern Indian patients and attempt to assess the overall mutation spectrum in India for detection of mutant allele for diagnostic purposes. Methods Patients from 109 unrelated families and their first-degree relatives comprising 400 individuals were enrolled in this study as part of an ongoing project. Genomic DNA was prepared from the peripheral blood of Indian WD patients. PCR was done to amplify the exons and flanking regions of the WD gene followed by sequencing, to identify the nucleotide variants. Results In addition to previous reports, we recently identified eight mutations including three novel (c.3412 + 1G > A, c.1771 G > A, c.3091 A > G) variants, and identified patients with variable phenotype despite similar mutation background suggesting potential role of modifier locus. Conclusions So far we have identified 17 mutations in eastern India including five common mutations that account for 44% of patients. Comparative study on WD mutations between different regions of India suggests high genetic heterogeneity and the absence of a single or a limited number of common founder mutations. Genotype–phenotype correlation revealed that no particular phenotype could be assigned to a particular mutation and even same set of mutations in different patients showed different phenotypes.     

Successive cambia development in Avicennia marina (Forssk.) Vierh. is not climatically driven in the seasonal climate at Gazi Bay, Kenya

This study is intended to provide early access to recent findings on the formation of the successive cambia of Avicennia marina (Forssk.) Vierh. in Kenya. The non-annual character of the growth layers was demonstrated by using three trees from a cambial marking experiment and three trees from a plantation of known age. The respective number of growth layers produced during one year was on average a half and three. Considering 28 stem disks of trees at three study sites, differing in local site conditions, growth layer development was shown to be strongly correlated with stem diameter (R²=0.84, p<0.0001, n=31). However, an additional influence of the site conditions was also demonstrated (homogeneity-of-slopes model test: F=54.72, p<0.0001, n=28). With increasing salinity and/or inundation class the width of the growth layers decreased. The significance of these variations in growth layer width offer interesting perspectives. The larger proportion of xylem in comparison with phloem in trees with wide as opposed to narrow growth layers may provide extra mechanical strength. On the other hand, the larger fraction of living tissue (phloem and parenchyma) in trees with thin growth layers may be beneficial for the water balance of the tree. Next to the non-annual nature of the growth layers and their networking pattern, more than one cambium was found to be simultaneously active. We conclude that classical dendrochronological methods (ring width measurements) should not be applied to A. marina (from Kenya).     

Effect of neurotrophins on differentiation, calcification and proliferation in cultures of human pulp cells

Neurotrophins (NTs) are expressed during tooth development. However, little is known about a role of NTs in differentiation of pulp cells into mineralizing cells. In this study, mRNA expressions of hard tissue-related proteins, calcification and proliferation are examined in cultures of human pulp (HP) cells. Nerve growth factor (NGF), brain derived neurotrophic factor (BDNF), neurotrophin (NT)-3 and NT-4/5 increased the mRNA levels of dentin sialophsphoprotein, alkaline phosphatase, osteopontin, type I collagen and bone morphogenetic protein-2 and mineral deposition in cultures of HP cells. The increased levels and manners varied, depending on the concentrations of NTs and hard-tissue related protein tested. On the other hand, only NGF significantly stimulated DNA synthesis in cultures of HP cells. These findings suggest that NTs characteristically regulate hard-tissue related protein expression, calcification and proliferation in pulp cells. NTs may accelerate pulp cell differentiation.     

Formation of filopodia in earthworm (Lumbricus terrestris) coelomocytes in response to osmotic stress

Coelomocytes, the immunodefense cells of the earthworm Lumbricus terrestris, are exposed to changing osmotic pressures as the worm's coelomic fluid responds to fluctuating wet-dry conditions of the surrounding soil. Using light and fluorescence microscopy combined with actin and tubulin disrupting drugs, we determined the effects of changing osmotic pressure on coelomocyte morphology. The coelomocytes from L. terrestris respond to an increase in environmental osmotic pressure from isotonic conditions (170 mOsm) to hypertonic conditions (715 mOsm) by changing from a round/petalloid morphology to a filopodial morphology. Cytoskeletal fluorescent staining studies indicate that for filopodia to form, the actin cortical ring, present in most coelomocytes in isotonic conditions, must be disrupted. Breakdown of the actin ring by exposure to a hypertonic environment or actin disrupting drugs allows the formation of actin or tubulin-based filopodia. The filopodia, or podial-like extensions formed by earthworm coelomocytes, may enable the cells to better explore their environment.     

A dynamic Brugada sign due to left anterior descending coronary artery occlusion

Brugada phenocopies (BrP) include several conditions with a common electrocardiographic (ECG) pattern that are indistinguishable from classical Brugada syndrome (BrS). In this report, we describe two cases of acute myocardial infarction (AMI) presenting as BrP. The majority of cases of BrP in AMI have been reported due to right coronary artery (RCA) occlusion. Rarely, the left anterior descending artery (LAD) is incriminated as the cause. In both our cases of BrP, LAD was the culprit vessel.     

宁夏河东沙地生物土壤结皮对土壤性质及入渗过程的影响

为了揭示沙漠化治理过程中生物土壤结皮覆盖对土壤入渗过程的影响规律,以宁夏河东沙地人工沙漠治理区4种典型地表覆盖类型：裸沙(BS)、藻类结皮(AC)、藓类结皮(MC)、草本-藓类结皮(H-MC)为研究对象,基于野外双环入渗试验与室内模拟,分析了3种生物土壤结皮覆盖下土壤性质的变化与土壤入渗特征。结果表明：(1)与BS相比,3种结皮覆盖下表层土壤砂粒含量减少2.0%-5.1%,粉粒含量增加3.6%-5.8%,有机质含量增加了5-6倍,AC和MC覆盖下土壤总孔隙度与饱和含水量降低,而H-MC与之相反;(2)平均入渗速率表现为BS>H-MC>AC>MC,1h累计入渗量表现为H-MC>BS>AC>MC,与BS相比,AC、MC和H-MC的初渗速率依次减少了14.3%、37.2%、11.8%,AC、MC的稳渗速率分别降低了14.4%和18.3%,H-MC的稳渗速率增加了4.5%;(3)三种模型中,Kostiakov模型最适用于模拟生物土壤结皮覆盖下土壤水分入渗过程。综上,研究区内不同发育程度生物土壤结皮改变了下层土壤的性质以及土壤的入渗特征,MC与AC阻碍水分入渗,H-MC促进水分入渗。