Nick Anthony’s research while affiliated with University of Arkansas at Fayetteville and other places

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Publications (9)


Comparing performance, morphological, physical, and chemical properties of eggs produced by 1940 Leghorn or a commercial 2016 Leghorn fed representative diets from 1940 or 2016
  • Article

July 2024

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15 Reads

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1 Citation

The Journal of Applied Poultry Research

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K.E. Anderson

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N. Anthony

Article Information Influence of Nutrition and Genetics on Bone Parameters of 1940 Leghorn and 2016 Commercial White Leghorns Description of Problem Humerus and Tibia Bones
  • Article
  • Full-text available

May 2024

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43 Reads

Bones play an essential role being responsible for the support of body mass, protection of internal organs, and providing musculature attachment sites while also serving as a reservoir for eggshell mineralization during the production phase. Targeted genetic selection has contributed to body morphometry and performance potential and could be inadvertently associated with undesirable effects on bone stability. In this study, bone parameters were compared between a contemporary and heritage line for the effect of strain and dietary regimen. A total of 320 White Leghorn laying hens (69 weeks of age) of two different strains were distributed into a 2 × 2 factorial arrangement creating 4 experimental treatment groups: 1). 2016 hen on 1940 diet, 2). 2016 hen on 2016 diet, 3). 1940 hen on 1940 diet, and 4). 1940 hen on 2016 diet with 8 replicates per treatment. Keel bones were assessed for deviations and or fractures. Significant differences (P ≥ 0.05) were observed for both deviations and/or fractures with the 2016 strain having more when compared to the 1940 strain. Humerus and tibia bones were analyzed for bone mineral density, breaking strength, and bone ash. Humerus and tibia weights which included both pre (with meat attached) and post weights (without meat attached) had significant differences (P ≤ 0.05) in the pre-weight in the 2016 hens, however no significant differences in the post weights. Results suggest that genetics played a role in the differences observed with the bone parameters measured and nutrition had few adverse effects.

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Figure 1. Alpha diversity of ceca and ileum. Alpha diversity metrics of ileum and ceca. Both ileum and ceca were significantly different but not among treatment groups.
Evaluating the Ileal and Cecal Microbiota Composition of a 1940 Heritage Genetic Line and a 2016 Commercial Line of white Leghorns Fed Representative Diets from 1940 to 2016

November 2023

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16 Reads

Applied Sciences

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Ramon D. Malherios

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[...]

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Nick Anthony

This study identified and evaluated differences between microbiome compositions of the ileum and ceca of 1940 and 2016 white leghorn genetic strains fed representative contemporary diets from those times. Ileal and cecal samples were collected at 69 weeks of age. Alpha and beta diversity metrics were generated, and the Analysis of Composition of Microbiomes (ANCOM) was utilized to determine significantly different taxa. Ileum and ceca alpha diversity were significantly different (p = 0.001; Q = 0.001); however, no differences between genetic lineage were observed (p > 0.05; Q > 0.05). Beta diversity between the ileum and ceca and the genetic lines was significantly different (p = 0.001; Q = 0.001). The ANCOM of the ileum showed significant differences between Proteobacteria and Actinobacteriota phyla (p ˂ 0.05) and significant differences between Pseudomonas, Rhizobiaceae, Leuconostoc, and Aeriscardovia genera (p ˂ 0.05). For ceca ANCOM, Proteobacteria, Firmicutes, Actinobacteriota, and Euryarchaeota phyla were significantly different (p ˂ 0.05), with Firmicutes having the highest relative abundance across all groups, and there were significant differences in genera Pseudomonas, Leuconostoc, Alloprevotella, and Aeri scardovia, with Alloprevotella having the highest relative abundance. The results suggest that genetic makeup in conjunction with the nutritional composition influences the cecal and ileal microbiota of corresponding hens.


Figure 2. Beta Diversity of Ceca and Ileum
Feed Ingredients and Mash 1 Diet Compositions
Evaluating the ileal and cecal microbiota composition of a 1940 heritage genetic line and a 2016 commercial line of white leghorns fed representative diets from 1940 and 2016

June 2023

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27 Reads

This study was conducted to identify and evaluate the differences between the microbiome composition of the ileum and ceca of 1940 and 2016 genetic strains of white leghorns fed representative contemporary diets from those times. Ileal and cecal samples were aseptically collected from both genetic lines at 69 weeks of age. The genomic DNA of the ileal and cecal contents were extracted and the V4 region of the 16S rDNA was sequenced on an Illumina Miseq. Microbiota data were filtered and aligned using the QIIME2 2020.2 pipeline. Alpha and beta diversity metrics were generated and the Analysis of Composition of Microbiomes (ANCOM) was utilized to determine significantly different taxa. Data were considered significant at P ≤ 0.05 for main effects and Q ≤ 0.05 for pairwise differences. Alpha diversity of the ileum and ceca were significantly different (P = 0.001; Q = 0.001; however, no differences between genetic lineage were observed (P > 0.05; Q > 0.05). The beta diversity between the ileum and ceca, as well as between the genetic lines (1940 vs. 2016) were significantly different from one another (P = 0.001; Q = 0.001). Using ANCOM, Proteobacteria and Actinobacteriota were significantly different than other phyla (P ˂ 0.05) with a higher relative abundance of Proteobacteria being observed among treatment groups 2 and 3, while Actinobacteriota had higher relative abundance in treatment groups 1 and 4. Among the significantly different genera in the ileum, Pseudomonas , Rhizobiaceae , Leuconostoc , and Aeriscardovia were different (P ˂ 0.05) with treatment groups 1 and 4 having a higher relative abundance of Aeriscardovia , while treatment groups 2 and 3 had higher relative abundance in both Pseudomonas and Leuconostoc . In the ceca, Proteobacteria , Firmicutes , Actinobacteriota , and Euryarchaeota were significantly different phyla (P ˂ 0.05) with Firmicutes having the highest relative abundance across all treatment groups. Among the significantly different genera ( Pseudomonas , Leuconostoc , Alloprevotella , and Aeriscardovia ), Alloprevotella had the highest relative abundance across all treatment groups 1 and 2, while Leuconostoc and Pseudomonas had the highest relative abundance in treatment group 4. Results from this study suggest that genetic makeup in conjunction with the nutritional composition of laying hens influences the cecal and ileal microbiota of corresponding hens.



Existence of pachytene piRNAs in chickens
a Roosters express pachytene piRNAs during spermatogenesis. (i) Key biological events during chicken spermatogenesis. (ii) Length distribution of total small RNAs. Ppm, parts per million. Blue, miRNAs. (iii) Abundance of piRNAs as measured by small RNA-seq. (iv) Expression of CIWI as measured by RNA-seq. Tpm, transcript per million. b Immunolabeling of squashed pachytene spermatocytes from adult chicken testes using anti-CIWI, anti-SYCP1, and DAPI. Scale bar, 10 µm. SYCP1, marker for synaptonemal complex formed during pachynema. We took at least 30 pictures and the representative pictures were shown. c Heatmap of normalized piRNA abundance per piRNA locus across the eight developmental stages of chicken testes. d Box plots of piRNA abundance at piRNA loci (n = 1321) in adult chicken testes and at their homolog regions (n = 637) in adult duck testes. Ppm: parts per million. Box plots show the 25th and 75th percentiles, whiskers represent the 5th and 95th percentiles, and midlines show median values. e Median value of (left) the mean phastCons score from 77 vertebrate genome alignments (probability that each nucleotide belongs to a conserved element) and (right) the mean phyloP score from 363 bird genome alignments (represent −log p-values under a null hypothesis of neutral evolution) of piRNA loci (red, n = 1321) and randomly shuffled control sequences (yellow, n = 10,000). Violin plots represent the medians of randomly shuffled control sequences that were computed 10,000 times.
Chicken piRNA loci are SV hotspots
a The landscape of SVs in chickens. (i) Quantity of each type of SV, (ii) density plots showing their length distributions, and (iii) pie chart showing their overlapping genomic regions. b Bar plots of the quantity of SVs in each chicken. INS, insertion; DEL, deletion; INV, inversion; DUP, tandem duplication. The Red Jungle Fowl had a significantly lower number of SVs compared to that of domesticated chickens (Z score = −15.9). Given our Red Jungle Fowl is from the same population selected for reference genome sequencing, the 4934 SVs detected in Red Jungle Fowl likely underrepresented the level of genetic diversity in the wild chicken population. c The number of SVs (red) and randomly shuffled control sequences (purple) falling into the piRNA loci (n = 1321). Violin plots represent the randomly shuffled control sequences that were computed 10,000 times.
Conserved mechanisms to achieve piRNA plasticity
a Example of a duplication overlapping with a piRNA locus and its piRNA abundance from two chicken individuals. Blue represents Watson strand mapping reads; Red represents Crick strand mapping reads. Ppm, parts per million. b Example of an inversion overlapping with two piRNA loci (cluster 1047 and cluster 1195) along with their nonoverlapping control piRNA loci (cluster 542 and cluster 377) and their piRNA abundance from two chicken individuals. Blue represents Watson strand mapping reads; Red represents Crick strand mapping reads. Ppm, parts per million. c Example of a duplication overlapping with a piRNA locus. (Left) piRNA abundance from two chicken individuals. Blue represents Watson strand mapping reads; Red represents Crick strand mapping reads. Ppm, parts per million. (Right) piRNA species abundance from the two chicken individuals that have read counts of 1 to 9. d Box plots of piRNA variance of Abundance (left), Strand bias (middle), and Shannon diversity index (right) among 23 chickens from 6 breeds. Box plots show the 25th and 75th percentiles, whiskers represent the 5th and 95th percentiles, and midlines show median values. e Number of human pachytene piRNA loci (red) and randomly shuffled control sequences (aquamarine) overlapping with SV hotspots within de novo pathogenic SVs detected in patients (left), healthy human populations (middle), and historical SVs in the common ancestor of humans and great apes (right). Violin plots represent the medians of randomly shuffled control sequences that were computed 10,000 times.
Convergent evolution drives the association between SV hotspots and pachytene piRNA loci
a Three models explain the association between pachytene piRNA loci and SV hotspots. b Number of pachytene piRNA loci (red) and randomly shuffled control sequences (magenta) overlapping with SDs from chickens (n = 861), humans (n = 3802), and mice (n = 659,775). Violin plots represent the medians of randomly shuffled control sequences that were computed 10,000 times. c Summary of diverse mutational mechanisms contributing to genetic instability at pachytene piRNA loci.
Silencing active TEs is a conserved function driving pachytene piRNA evolution
a The percentage of active TE sequences and total TE sequences in piRNA loci (red) and in randomly shuffled control sequences (aquamarine). Human n = 88, and mouse n = 100. Violin plots represent 10,000 randomly shuffled control sequences. b Immunofluorescence labeling of mouse round spermatids. γH2AX, marker for double strand breaks. The foci numbers were quantified from 90 round spermatids from three biological replicates. Scale bar, 10 µm. c Scatter plot of mean TE transcript abundance in Mov10l1 CKO mutants versus that of littermate controls (n = 3). Each filled circle represents a TE family. Red, q value < 0.1. Each large circle represents an active TE family. Tpm transcript per million. d The 5′-5′ overlap between sense and anti-sense piRNAs mapping to TEs that are significantly increased in Mov10l1 CKO mutants. Data are mean ± standard deviation (n = 3). Ppm parts per million. e Scatter plot of mean TE transcript abundance in 19 chickens from the 6 breeds versus mean TE piRNA abundance in 23 chickens from the 6 breeds. 30 active TE families (red). Rpkm reads per kilobase pair per million reads mapped to the genome. p value was calculated by Spearman’s rank correlation coefficient statistical test. f Box plots of the distance between SV hotspots and nearest protein coding genes in (upper) humans (piRNA n = 88, SV minus piRNA n = 269) and in (lower) chicken macrochromosomes (piRNA n = 779, SV minus piRNA n = 26). We only calculated the distance on macrochromosomes including chromosome Z in chickens where most of the piRNA loci localized (751/1321) because the assembly of microchromosomes has not been completed. p value is smaller than the threshold we can compute. Box plots show the 25th and 75th percentiles, whiskers represent the 5th and 95th percentiles, and midlines show median values. g Example of a pachytene piRNA locus overlapping with 16 SVs deposited in ClinVar. From top to bottom: RefSeq, pathogenic SVs (each SV is labeled by its Variation ID, and Red is associated with autism spectrum disorder), and piRNA reads from adult human testes (Blue represents Watson strand mapping reads; Red represents Crick strand mapping reads).
Amniotes co-opt intrinsic genetic instability to protect germ-line genome integrity

February 2023

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279 Reads

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7 Citations

Unlike PIWI-interacting RNA (piRNA) in other species that mostly target transposable elements (TEs), >80% of piRNAs in adult mammalian testes lack obvious targets. However, mammalian piRNA sequences and piRNA-producing loci evolve more rapidly than the rest of the genome for unknown reasons. Here, through comparative studies of chickens, ducks, mice, and humans, as well as long-read nanopore sequencing on diverse chicken breeds, we find that piRNA loci across amniotes experience: (1) a high local mutation rate of structural variations (SVs, mutations ≥ 50 bp in size); (2) positive selection to suppress young and actively mobilizing TEs commencing at the pachytene stage of meiosis during germ cell development; and (3) negative selection to purge deleterious SV hotspots. Our results indicate that genetic instability at pachytene piRNA loci, while producing certain pathogenic SVs, also protects genome integrity against TE mobilization by driving the formation of rapid-evolving piRNA sequences.


Fourth Report on Chicken Genes and Chromosomes 2022

January 2023

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1,094 Reads

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20 Citations

Cytogenetic and Genome Research

The chicken continues to hold its position as a leading model organism within many areas of research, as well as a being major source of protein for human consumption. The First Report on Chicken Genes and Chromosomes [Schmid et al., 2000], which was published in 2000, was the brainchild of the late, and sadly missed, Prof Michael Schmid of the University of Würzburg. It was a publication bringing together updates on the latest research and resources in chicken genomics and cytogenetics. The success of this First report led to the subsequent publication of the Second [Schmid et al., 2005] and Third [Schmid et al., 2015] reports proving popular references for the research community. It is now our pleasure to be able to introduce publication of the Fourth report. Being seven years since the last report, this publication captures the many advances that have taken place during that time. This includes presentation of the detailed genomic resources that are now available, largely due to increasing capabilities of sequencing technologies and which herald the pangenomic age, allowing for a much richer and more complete knowledge of the avian genome. Ongoing cytogenetic work also allows for examination of chromosomes, specific elements within chromosomes and the evolutionary history and comparison of karyotypes. We also examine chicken research efforts with a much more ‘global’ outlook with a greater impact on food security and the impact of climate change, and highlight the efforts of international consortia, such as the Chicken Diversity Consortium. We dedicate this Report to Michael.


Citations (3)


... До голяма степен това влияние се обуславя от процеса на хибридизация, чийто ефект е най-силно изразен при четирилинейния хибрид "ISA-Brown", където стойностите на тези показатели са най-високи (43,27 mm; 78,90 ). Това се дължи на силно изразен хетерозисен ефект, повлияващ количествено и качествено яйценосната продук-тивност на птиците (Buzała et al., 2015;Heflin et al., 2018;Dannica et al., 2024). ...

Reference:

Comparative study on the morphological traits and hatchability of egg from dual purpose and laying purebred lines and crosses
Comparing performance, morphological, physical, and chemical properties of eggs produced by 1940 Leghorn or a commercial 2016 Leghorn fed representative diets from 1940 or 2016
  • Citing Article
  • July 2024

The Journal of Applied Poultry Research

... Several full-length LTR containing elements also remain intact in the chicken genome and possess potentially functional ORFs ( 147 ). Thus, CR1 and LTR containing elements should be targeted by piRNAs to prevent their spreading ( 148 ). In addition, since many genes carry fragments of retrotransposable elements within their introns, piRNAs may be attracted to introns and promote intron removal during nascent pre-mRNA splicing ( 149 ). ...

Amniotes co-opt intrinsic genetic instability to protect germ-line genome integrity

... Since the release of the initial draft genome assembly (galgal2) of an RJF individual 2 , multiple improved assemblies (galgal3-galgal5 and GRCg6a) have been developed 3,4 . More recently, the Vertebrate Genomes Project (VGP) also assembled pseudo-haplotype genomes (GRCg7b and GRCg7w) of a hybrid individual from a broiler mother and a layer father using long sequencing reads and multiple scaffolding data 5,6 . There are also several assemblies for indigenous chickens deposited in GenBank, such as Yeonsan Ogye chicken (Ogye1.0) ...

Fourth Report on Chicken Genes and Chromosomes 2022

Cytogenetic and Genome Research