[Magnetic resonance imaging analysis on the effect of marathon on the dynamic changes of morphological characteristics of patellar tendon in amateur marathon runners after half marathon].

Objective: To explore the dynamic changes of morphological characteristics of patellar tendon (PT) in amateur athletes after a half-marathon using magnetic resonance imaging. Methods: A total of 19 male amateur marathon runners with 38 knee joints,aged from 26 to 53(41.5±7.9) years, running for 3 to 18 years, with a weekly running volume of 30-90 km and a monthly running volume of 100-300 km were enrolled and underwent 1.5T MRI scan before the half-marathon, within 3 hours after running and 3 days after running. Ten healthy male volunteers with 20 knee joints, who had never participated in marathon and exercised (including but not limited to running) per week for less than 150 minutes were recruited as the control group, aged from 26 to 54 (39.4±9.1) years. Firstly, the PT signal was qualitatively assessed on fat-suppressed proton density-weighted imaging (fs-PDWI) sequence to observe the presence of patellar tendinitis. Then, the length, proximal, middle and distal cross-sectional area (CSA), and volume of PT were measured using the post-processing tool ITK-SNAP, and the data were standardized. The independent sample t-test was used for comparing. One-way repeated measures analysis of variance was used to analyze the morphological changes of PT before and after half-marathon running. Results: The incidence of asymptomatic patellar tendinitis in amateur marathon runners was 26.3% (5/19). No significant MR signal changes of PT were observed in all runners after running. The proximal CSA in runners group was larger than that in controls [(4.20±0.62) mm2/kg3/4 vs (3.63±0.57) mm2/kg3/4, P<0.05], and there was no significant difference in length, medium and distal CSA and volume(all P>0.05). The length, proximal and distal CSA and volume of PT in runners group increased at 3 h after running [(47.35±3.22) mm vs (46.83±3.35) mm; (102.52±13.03) mm2 vs (98.98±13.14) mm2; (108.67±15.72) mm2 vs (100.27±14.37) mm2; (4 020.36±514.38) mm3 vs (3 826.57±499.23) mm3, all P<0.05]. There was no significant difference between before running and 3 days after running(all P>0.05). The middle CSA were not significantly different among different periods(all P>0.05). Conclusion: Marathon has effect on the normal PT morphology in male amateur marathon runners, showing an increase in proximal CSA. A half-marathon will cause reversible changes in PT length, regional CSA and volume.

A combined approach using slightly acidic electrolyzed water and UV exposure to improve egg internal quality during storage.

This study investigated the combined efficacy of slightly acidic electrolyzed water (SAEW) and UV light (UV) in improving egg internal quality (weight loss, Haugh unit, yolk index, albumen pH) over a 6-wk storage time at 25°C. Eggs were preserved after immersion for 4 min in SAEW (30 mg/L), irradiation for 4 min under a UV lamp, or a combination of SAEW and UV treatment for 4 min. The combination of SAEW and UV inhibited the deterioration of yolk index over the storage period, as well as reducing the extent of decrease in Haugh unit and of weight loss during storage at 25°C, and it was more effective than SAEW or UV alone in maintaining egg internal quality (P < 0.05). The results highlight the promising use of a SAEW and UV combination treatment to improve egg internal quality during storage.

Comprehensive analysis of miRNAs, lncRNAs, and mRNAs reveals potential players of sexually dimorphic and left-right asymmetry in chicken gonad during gonadal differentiation.

Despite thousands of sex-biased genes being found in chickens, the genetic control of sexually dimorphic and left-right asymmetry during gonadal differentiation is not yet completely understood. This study aimed to identify microRNAs (miRNAs), long noncoding RNAs (lncRNAs), messenger RNAs (mRNAs), and signaling pathways during gonadal differentiation in chick embryos (day 6/stage 29). The left and right gonads were collected for RNA sequencing. Sex-biased, side-biased miRNAs, lncRNAs, mRNAs, and shared differentially expressed miRNAs (DEmiRNA)-differentially expressed mRNAs (DEmRNA)-differentially expressed lncRNAs (DElncRNA) interaction networks were performed. A total of 8 DEmiRNAs, 183 DElncRNAs, and 123 DEmRNAs were identified for the sex-biased genes, and 7 DEmiRNAs, 189 DElncRNAs, and 183 DEmRNAs for the side-biased genes. The results of quantitative real-time PCR were generally consistent with the RNA-sequencing results. The study suggested that miRNAs and lncRNAs regulation were novel gene-specific dosage compensation mechanism and they could contribute to left-right asymmetry of chicken, but sex-biased and side-biased miRNAs, lncRNAs, and mRNAs were independent of each other. The competing endogenous RNA (ceRNA) networks showed that 17 target pairs including miR-7b (CYP19A1, FSHR, GREB1, STK31, CORIN, and TDRD9), miR-211 (FSHR, GREB1, STK31, CORIN, and TDRD9), miR-204 (FSHR, GREB1, CORIN, and TDRD9), and miR-302b-5p (CYP19A1 and TDRD9) may play crucial roles in ovarian development. These analyses provide new clues to uncover molecular mechanisms and signaling networks of ovarian development.

Application of slightly acidic electrolyzed water and ultraviolet light for Salmonella enteritidis decontamination of cell suspensions and surfaces of artificially inoculated plastic poultry transport coops and other facility surfaces.

The efficiency of combination treatment of slightly acidic electrolyzed water (SAEW) and ultraviolet light (UV) for inactivation of Salmonella enteritidis (S. enteritidis) on the surface of plastic poultry coops and other facility surfaces was evaluated in the presence of organic matter. The bactericidal activities of SAEW, UV + SAEW, and composite phenol (CP) for inactivating S. enteritidis were also compared. Moreover, a model of UV + SAEW treatment of plastic transport coops with different times and available chlorine concentrations (ACC) was developed using multiple linear regression analysis. There are differences between SAEW and CP inactivation of S. enteritidis on coops, stainless steel, and glass surfaces (P < 0.05), and there are no differences between SAEW and CP on tire surfaces (P > 0.05). Disinfection of some rough material surfaces with SAEW treatment alone under feces interference on poultry farms may need a longer treatment time and/or a higher ACC than smooth surfaces. The combined treatment of UV and SAEW showed higher inactivation efficiency of S. enteritidis compared to CP and SAEW treatment alone (P < 0.05) in pure cultures or on the facility surfaces. A complete 100% inactivation of S. enteritidis on plastic poultry coop surfaces was obtained by using UV + SAEW with an ACC of 90 mg/L for more than 70 s. The established model had a good fit that was quantified by the determination coefficient R2 (0.93) and a lack of fit test (P > 0.05). The bactericidal efficiency of UV + SAEW increased with greater ACC and increasing time. The findings of this study indicate that the combination treatment of UV and SAEW may be a promising disinfection method and could be used instead of SAEW alone, especially on rough materials in the presence of organic matter on poultry farms.

The synergistic effects of slightly acidic electrolyzed water and UV-C light on the inactivation of Salmonella enteritidis on contaminated eggshells.

Salmonella enteritidis (S. enteritidis) infection has been recognized as one of the most common bacterial causes of human gastroenteritis worldwide and is closely associated with eggs. Slightly acidic electrolyzed water (SAEW) is an emerging environmentally friendly technology for disinfecting eggshell surfaces to remove dirt and pathogenic microorganisms. However, the efficiency of SAEW could be affected by the presence of manure. UV-based advanced oxidation processes have been studied to improve the microorganism's inactivation effect of disinfection. Therefore, in this study, the synergistic bactericidal efficacy of SAEW and UV-C light (ultraviolet lamp, λ = 254 nm) for inactivation of S. enteritidis on artificially inoculated eggshells with or without manure was evaluated, and the bactericidal efficacy of different combination treatments of SAEW and UV-C light was compared. Without manure interference, complete inactivation (reduction of 6.54 log10 CFU/g) of S. enteritidis on the surface of eggshells was achieved following a 4-min treatment with SAEW+UV at an available chlorine concentration (ACC) of 20 mg/L. In the presence of manure, a 3.02 log reduction was achieved following a 4-min treatment with SAEW+UV at an ACC of 30 mg/L. Simultaneous treatment with SAEW and UV light exhibits higher bactericidal activity for eggshells than other combination process methods with UV and SAEW. The results suggest that the combined treatment of SAEW+UV is a novel method to enhance the microbial safety of eggshells.

Association of host genetics with intestinal microbial relevant to body weight in a chicken F2 resource population.

Host-microbiota interactions describe a co-evolution and mutualistic symbiosis. Gut microbial communities are important for diverse host functions. However, in birds, the relationship between the composition of the intestinal microbiota and the genetic variation of the host is not clearly understood. To dissect these interactions, a Chinese yellow broiler line (genetically selected for a high growth rate) and Huiyang Beard chickens (low growth rate) were crossed, generating an F2 population. The population structures of the gut microbes in the phenotypically high and low 91-d body weight individuals of both sexes in the F2 population were studied. Interestingly, a non-metric multidimensional scaling analysis revealed that the microbiota of the high-weight and low-weight females was clearly separated into 2 clusters. A ß-diversity analysis showed that the locus rs16775833 within the doublesex and mab-3-related transcription factor (DMRT) gene cluster accounted for approximately 21% of the variation in the population structure of the gut microbiota. Furthermore, the 2 genetic loci rs15142709 and rs15142674 were significantly associated with specific species of Methanobacterium. These loci are located in the pleiomorphic adenoma gene 1 (PLAG1) and lck/yes-related novel tyrosine kinase (LYN) genes, which are involved in cell differentiation and growth. This finding suggests evidence for the influence of the host genetics on the composition of the gut microbiota in birds and the importance and utility of the host-microbe status to better understand its effect on the potential growth of birds.

Efficacy of slightly acidic electrolyzed water on the microbial safety and shelf life of shelled eggs.

This study evaluated the efficacy of slightly acidic electrolyzed water (SAEW) for the inactivation of Salmonella Enteritidis and Escherichia coli on shelled eggs as well as studied the shelf life and internal quality attributes (i.e., weight loss, Haugh unit, yolk index, albumen pH, and yolk pH) of eggs during storage at 25°C. The decontamination test egg samples (freshly laid) were inoculated and immersed for 1, 2, 3, and 4 min in each treatment (i.e., SAEW, acidic electrolyzed water, NaClO solution, and sterile deionized water) at available chlorine concentrations (ACCs) of 10, 18 and 26 mg/L. The storage test eggs (freshly laid) were immersed for 3 min in 4 treatments (i.e., SAEW, acidic electrolyzed water, NaClO solution, and no treatment) and stored for 30 D at 25°C. The effects of the SAEW on microbiological qualities and freshness parameters were investigated. Decontamination of eggs with SAEW showed an equivalent or higher bactericidal effect compared to other treatments. A complete inactivation of S. Enteritidis and E. coli on the surface of shelled egg samples resulted from treatment with SAEW at an ACC of 26 mg/L for 3 and 4 min, respectively. Almost all of the egg quality parameters investigated in the present study were significantly (P < 0.05) affected by the storage time. As storage time increased, the yolk index and Haugh unit value decreased, and weight loss, albumen pH, and yolk pH increased. However, SAEW treatment minimized weight loss (5.52%) and preserved the albumen and yolk quality better than no treatment at 25°C. Relative to acidic electrolyzed water and NaClO solution, the advantages of SAEW are reduced corrosion of egg surfaces and potentially less water and CO2 escaping from eggshell pores. The results highlight the promising use of SAEW to enhance the microbial safety and to extend the shelf life of shelled eggs. Future combined methods with SAEW and other treatments are also needed.

Effects of sodium butyrate on intestinal health and gut microbiota composition during intestinal inflammation progression in broilers.

Butyric acid is a beneficial feed additive used in animal production, including poultry production. However, there are few reports on butyric acid as a prophylactic treatment against intestinal inflammation in broilers. The current study explored the effect of sodium butyrate (SB) as a prophylactic treatment on the intestinal health and gut microbiota of broilers with intestinal inflammation induced by dextran sulfate sodium (DSS) by monitoring changes in intestinal histopathology, gut leakiness indicators, inflammatory cytokines, and gut microbiota composition. Sodium butyrate supplementation prior to DSS administration significantly reduced the lesion scores of intestinal bleeding (P < 0.05) and increased villus height and the total mucosa of the ileum (P < 0.05). Regardless of intestinal inflammation, supplementation with SB at 300 mg/kg significantly decreased the levels of D (-)-lactate (P < 0.05), interleukin-6, and interleukin-1ß (P < 0.05) but increased the level of interleukin-10 (P < 0.05). The SB treatment did not affect the alpha diversity of intestinal microbiota during intestinal inflammation progression but altered their composition, and the microbial community structure of treated broilers was similar to that of control broilers. Taken together, our results reveal the importance of SB in improving intestinal development, inducing an anti-inflammatory effect during intestinal inflammation progression, and modulating the microbial community in broilers. Sodium butyrate seems to be optimized for anti-inflammatory effects at higher doses (300 mg/kg SB).

Associations between variants of bone morphogenetic protein 7 gene and growth traits in chickens.

1. Enhancing bone strength to solve leg disorders in poultry has become an important goal in broiler production. Bone morphogenetic protein 7 (BMP7), a member of the BMP family, represents an attractive therapeutic target for bone regeneration in humans and plays critical roles in skeletal development. 2. The objective of this study was to investigate the relationship between BMP7 gene expression, single-nucleotide polymorphisms (SNPs) and growth traits in chickens. Here, a SNP (c.1995T>C) in the chicken (Gallus gallus) BMP7 gene was identified, that was associated with growth and carcass traits. 3. Genotyping revealed that the T allele occurred more frequently in breeds with high growth rates, whereas the C allele was predominant in those with low growth rates. The expression level of BMP7 in the thigh bone of birds with the TT genotype was significantly higher than in those with the CC genotype at 21, 42 and 91 d of age. 4. These findings suggest that selecting the birds with the TT genotype of SNP c.1995T>C could improve bone growth, could reduce leg disorders in fast-growing birds. The SNP c.1995T>C may serve as a selective marker for improving bone growth and increasing the consistency of body weights in poultry breeding.

Unidirectional Spin-Wave-Propagation-Induced Seebeck Voltage in a PEDOT:PSS/YIG Bilayer.

We clarify the physical origin of the dc voltage generation in a bilayer of a conducting polymer film and a micrometer-thick magnetic insulator Y_{3}Fe_{5}O_{12} (YIG) film under ferromagnetic resonance and/or spin wave excitation conditions. The previous attributed mechanism, the inverse spin Hall effect in the polymer [Nat. Mater. 12, 622 (2013)NMAACR1476-112210.1038/nmat3634], is excluded by two control experiments. We find an in-plane temperature gradient in YIG which has the same angular dependence with the generated voltage. Both vanish when the YIG thickness is reduced to a few nanometers. Thus, we argue that the dc voltage is governed by the Seebeck effect in the polymer, where the temperature gradient is created by the nonreciprocal magnetostatic surface spin wave propagation in YIG.

Dextran sulphate sodium (DSS) causes intestinal histopathology and inflammatory changes consistent with increased gut leakiness in chickens.

1. The clinical severity, histological changes, indicators of gut leakiness and inflammatory cytokine profiles were studied in chickens with dextran sulphate sodium (DSS)-induced intestinal inflammation. 2. The experimental groups (1.25%, 1.5% and 2.5% DSS) showed clinical signs, such as loose stools and weight loss, which increased with additional treatment days and, as expected, the effects of DSS-induced intestinal inflammation were time and dose-dependent. 3. After 10 d, histological manifestations were evident, including goblet cell depletion, mucus layer loss, significantly shorter villi and a thinner total ileal mucosa. 4. The d(-)-lactate value, which was used as a gut leakiness indicator, was significantly increased in the 2.5% DSS group. 5. Expression of the inflammatory cytokines interleukin-1Beta, tumour necrosis factor alpha and interleukin-10 in the serum significantly increased with DSS treatment. 6. This study indicates that the experimental intestinal inflammation induced by DSS is an ideal model to study the pathogenic mechanisms of intestinal inflammation in chickens and to test the efficacy of therapies.

Design and performance of an X-ray scanning microscope at the Hard X-ray Nanoprobe beamline of NSLS-II.

A hard X-ray scanning microscope installed at the Hard X-ray Nanoprobe beamline of the National Synchrotron Light Source II has been designed, constructed and commissioned. The microscope relies on a compact, high stiffness, low heat dissipation approach and utilizes two types of nanofocusing optics. It is capable of imaging with ∼15 nm × 15 nm spatial resolution using multilayer Laue lenses and 25 nm × 26 nm resolution using zone plates. Fluorescence, diffraction, absorption, differential phase contrast, ptychography and tomography are available as experimental techniques. The microscope is also equipped with a temperature regulation system which allows the temperature of a sample to be varied in the range between 90 K and 1000 K. The constructed instrument is open for general users and offers its capabilities to the material science, battery research and bioscience communities.

Improvement of iturin A production in Bacillus subtilis ZK0 by overexpression of the comA and sigA genes.

Bacillus subtilis ZK0, which was isolated from cotton, produces a type of lipopeptide antibiotic iturin A that inhibits the growth of pathogenic fungi on agricultural crops. However, the low level of iturin A production by B. subtilis ZK0 does not support its large-scale application. In this study, B. subtilis ZK0 was subjected to genetic manipulation to improve iturin A production. By the independent or simultaneous overexpression of two regulatory genes (comA and sigA), iturin A production by B. subtilis ZK0 was significantly increased. When both genes were simultaneously overexpressed under optimal conditions, iturin A production increased up to 215 mg l-1 (an approximate 43-fold increase compared with B. subtilis ZK0). Moreover, overexpression of both genes was unexpectedly found to inhibit biofilm formation by B. subtilis ZK0, indicating that the phenomenon of 'stuck fermentation' would be avoided during B. subtilis ZK0 fermentation. In conclusion, a genetic manipulation method that improves iturin A production and inhibits biofilm formation in B. subtilis ZK0 is reported for the first time and this method has the potential to be widely applied in B. subtilis ZK0 commercial fermentation. SIGNIFICANCE AND IMPACT OF THE STUDY: This study provides new perspectives on improving iturin A production by Bacillus subtilis. Our newly engineered strains could be applied to commercial fermentation by enhancing yields of iturin A and reducing the rate of 'stuck fermentation'. Increased production would facilitate more widespread application of this powerful antibiotic.

Association of single nucleotide polymorphisms in the microRNA miR-1596 locus with residual feed intake in chickens.

MicroRNAs are an abundant class of small non-coding RNAs that regulate gene expression. Genetic variations in microRNA sequences may be associated with phenotype differences by influencing the expression of microRNAs and/or their targets. This study identified two single nucleotide polymorphisms (SNPs) in the genomic region of the microRNA miR-1596 locus of chicken. Of the two SNPs, one was 95 bp upstream of miR-1596 (g.5678784A>T) and the other was in the middle of the sequence producing the mature microRNA gga-miR-1596-3p (g.5678944A>G). Genotypic distribution of the two SNPs had large differences among 12 chicken breeds (lines), especially between the fast-growing commercial lines and the slow-growing Chinese indigenous breeds for the g.5678784A>T SNP. Only the g.5678784A>T SNP was significantly associated with residual feed intake (RFI) in the F2 population derived from a fast-growing and a slow-growing broiler as well as in the pure Huiyang bearded chicken. The birds with the AA genotype of the g.5678784A>T SNP had lower RFI and higher expression of the mature gga-miR-1596-3p microRNA of miR-1596 than did those with the other genotypes of the same SNP. We also found that the expression of the mature gga-miR-1596-3p microRNA of miR-1596 was significantly associated with RFI. These findings suggest that miR-1596 can become a candidate gene related to RFI, and its genetic variation may contribute to changes in RFI by altering expression levels of the mature gga-miR-1596-3p microRNA in chicken.

Lycopene reduces the negative effects induced by lipopolysaccharide in breeding hens.

This study assessed the effects of lycopene on the antioxidant capacity, biochemical parameters, and immune organ index of breeding hens following lipopolysaccharide (LPS) injection. The study had a 2 × 2 factorial design. Healthy Xing-hua breeding hens (720) were divided into groups with 6 replicates per group and 30 hens per replicate. Hens were fed on a rice-soya bean basal meal supplemented with different levels of lycopene (0, 20, 40, or 80 mg/kg diet). On d 35, two hens from each replicate were injected subcutaneously with 1 mg/kg body weight of either LPS or sterile saline (control group). Blood samples were collected at 0, 6, and 24 h post-injection. At 24 h post-injection, hens were sacrificed and the thymus, spleen, and bursa of Fabricius were removed. The results revealed that LPS significantly decreased high density lipoprotein cholesterol (HDLC), triidothyronine (T3), reduced glutathione to oxidised glutathione ratio (GSH/GSSG), thymus and bursal indexes, and increased low density lipoprotein cholesterol (LDLC). Lycopene supplementation significantly increased HDLC, T3, GSH/GSSG, and immune organ index, and decreased total cholesterol, LDLC, and blood urea nitrogen (BUN). The interaction effects of lycopene and LPS were significant on BUN and T3. Lycopene supplementation affected inflammatory immune response based on increased immune organ index of breeding hens by relieving the LPS-induced stress.

Single nucleotide polymorphism variants within tva and tvb receptor genes in Chinese chickens.

Avian leukosis is an immunosuppressive neoplastic disease caused by avian leukosis viruses (ALV), which causes tremendous economic losses in the worldwide poultry industry. The susceptibility or resistance of chicken cells to subgroup A ALV and subgroup B, D, and E ALV are determined by the receptor genes tumor virus locus A (tva) and tumor virus locus B (tvb), respectively. Four genetic resistant loci (tva(r1), tva(r2), tva(r3), and tva(r4)) in tva receptor gene and a genetic resistant locus tvb(r) in the tvb receptor gene have been identified in inbred lines of White Leghorn. To evaluate the genetic resistance to subgroup A, B, D, and E ALV, genetic variations within resistant loci in tva and tvb genes were screened in Chinese local chicken breeds and commercial broiler lines. Here, the heterozygote tva(s1/r1) and the resistant genotype tva(r2/r2), tva(r3/r3), and tva(r4/r4) were detected in Chinese chickens by direct sequencing. The heterozygote tva(s1/r1) was detected in Huiyang Bearded chicken (HYBC), Rizhaoma chicken, and commercial broiler line 13 to 15 (CB13 to CB15), with the frequencies at 0.08, 0.18, 0.17, 0.25, and 0.15, respectively. The resistant genotype tva(r2/r2) was detected in Jiningbairi chicken (JNBRC), HYBC, and CB15, with the frequencies at 0.03, 0.08, and 0.06, respectively, whereas tva(r3/r3) and tva(r4/r4) were detected in 19 and 17 of the 25 Chinese chickens tested, with the average frequencies at 0.13 and 0.20, respectively. Furthermore, the resistant genotype tvb(r/r) was detected in JNBRC, CB07, CB12, CB14, and CB15 by pyrosequencing assay, with the frequencies at 0.03, 0.03, 0.11, 0.09, and 0.15, respectively. These results demonstrated that the potential for genetic improvement of resistance to subgroup A, B, D, and E ALV were great both in Chinese local chickens and commercial broilers. This study provides valuable insight into the selective breeding for chickens genetically resistant to ALV.

An early Cambrian agglutinated tubular lophophorate with brachiopod characters.

The morphological disparity of lophotrochozoan phyla makes it difficult to predict the morphology of the last common ancestor. Only fossils of stem groups can help discover the morphological transitions that occurred along the roots of these phyla. Here, we describe a tubular fossil Yuganotheca elegans gen. et sp. nov. from the Cambrian (Stage 3) Chengjiang Lagerstätte (Yunnan, China) that exhibits an unusual combination of phoronid, brachiopod and tommotiid (Cambrian problematica) characters, notably a pair of agglutinated valves, enclosing a horseshoe-shaped lophophore, supported by a lower bipartite tubular attachment structure with a long pedicle with coelomic space. The terminal bulb of the pedicle provided anchorage in soft sediment. The discovery has important implications for the early evolution of lophotrochozoans, suggesting rooting of brachiopods into the sessile lophotrochozoans and the origination of their bivalved bauplan preceding the biomineralization of shell valves in crown brachiopods.

Accumulation of melanin in the peritoneum causes black abdomens in broilers.

A suspected case of localized visceral hyperpigmentation was described for a breed of broiler in China. Using optical microscopy, the accumulation of pigments in the abdominal skin and visceral peritoneum was observed. Electron microscopy was used to further study the ultrastructure of the pigmented peritoneum, and pigment granules resembling melanosomes at different stages were found, and melanocytes were present in this tissue. Infrared spectroscopy was used to analyze the physical-chemical properties of pigments extracted from these broilers. Using synthetic melanin as a reference and the melanin from the peritoneum of Silkie fowls as a control, the pigments in the peritonea of these broilers were found to be melanin, and it had a chemical structure similar to that of melanin from the Silkie fowl peritoneum. In this way, the black abdomens of these broilers were found to have been caused by accumulation of melanin produced by melanocytes in visceral peritonea.

An association between genetic variation in the roundabout, axon guidance receptor, homolog 2 gene and immunity traits in chickens.

The roundabout, axon guidance receptor, homolog 2 (ROBO2) gene is one member of the roundabout (ROBO) family, which belongs to the immunoglobulin superfamily. The ROBO molecules are known to function in axon guidance and cell migration and are involved in SLIT/ROBO signaling. In this study, we obtained the full-length cDNA sequence of the chicken ROBO2 gene. Sequence analysis indicated that 3 SNP (1418G > A, 1421C > A and 2462T > C) exist in exons 5 and 12 of the ROBO2 gene. Genotyping results revealed that the allele frequency of SNP 1421C > A was similar in all tested breeds, but the allele frequencies of the other 2 SNP were different between White Leghorn and Chinese indigenous chickens. Allele G of 1418G > A and allele T of 2462T > C predominated in the Chinese indigenous breed, whereas alleles A and C predominated in the White Leghorn breed. Association analyses revealed that birds with the GG genotype of SNP 1418G > A or the TT genotype of SNP 2462T > C had significantly higher antibody responses to Newcastle disease virus (NDV_S/P; P < 0.01) than carriers of the A allele (GA and AA) or the C allele (TC), respectively. Real-time PCR further revealed that ROBO2 expression in the spleens of the birds with higher antibody responses (GG and TT genotypes at SNP 1418 and 2462, respectively) was significantly higher than in the spleens of birds with the AA and AG genotypes at SNP 1418 or the TC genotype at SNP 2462 (P < 0.01). The results demonstrated that genetic variation at the ROBO2 gene plays a key role in the immune response to Newcastle disease virus, and SNP 1418G > A and 2462T > C can be used as genetic markers for the selection of chickens with stronger immune responses to Newcastle disease virus.

The Bionanoprobe: hard X-ray fluorescence nanoprobe with cryogenic capabilities.

Hard X-ray fluorescence microscopy is one of the most sensitive techniques for performing trace elemental analysis of biological samples such as whole cells and tissues. Conventional sample preparation methods usually involve dehydration, which removes cellular water and may consequently cause structural collapse, or invasive processes such as embedding. Radiation-induced artifacts may also become an issue, particularly as the spatial resolution increases beyond the sub-micrometer scale. To allow imaging under hydrated conditions, close to the `natural state', as well as to reduce structural radiation damage, the Bionanoprobe (BNP) has been developed, a hard X-ray fluorescence nanoprobe with cryogenic sample environment and cryo transfer capabilities, dedicated to studying trace elements in frozen-hydrated biological systems. The BNP is installed at an undulator beamline at sector 21 of the Advanced Photon Source. It provides a spatial resolution of 30 nm for two-dimensional fluorescence imaging. In this first demonstration the instrument design and motion control principles are described, the instrument performance is quantified, and the first results obtained with the BNP on frozen-hydrated whole cells are reported.