Embryonic thermal manipulation of poultry birds: Lucrative and deleterious effects.

The major efforts to improve feed conversion, increase the body weight and breast muscle yield of broilers have been focused on feeding and management at the post hatch period. However, incubation temperature is the most significant factor for the egg hatching rate, chick quality, and post hatch performance. Therefore, incubation factors affecting the performance should be taken with necessary precautions. Incubation temperature not only affects the early development of the hatchlings but also has a lasting impact on the characteristics of the chicks, such as final body weight and meat quality traits. This article provides an overview about embryonic thermal manipulation (TM) of domestic fowls and review the lucrative and deleterious effects of embryonic TM on embryo development, muscle growth, thermotolerance acquisition, and immunity.

Morphometric studies on the appendicular bony skeleton of the ostriches (Struthio Camelus).

BACKGROUND: Morphometric study of the bony elements of the appendicular skeleton in the ostrich was fully described and identified. The appendicular skeleton included the bones of the pectoral girdle, the wing, the pelvic girdle and the pelvic limb. RESULTS: The shoulder girdle of the ostrich included the scapula and coracoid bones. The scapula appeared as a flattened spoon-like structure. The coracoid bone appeared quadrilateral in outline. The mean length of the scapula and coracoid (sternal wing) were 15.00 ± 0.23 and 10.00 ± 0.17 cm, respectively. The wing included the humerus, ulna, radius, radial carpal bone, ulnar carpal bone, carpometacarpus and phalanges of three digits. The mean length of the humerus, radius, and ulna were 33.00 ± 0.46, 10.50 ± 0.40 and 11.50 ± 0.29 cm respectively. The carpometacarpus was formed by the fusion of the distal row of carpal bones and three metacarpal bones. Digits of the wing were three in number; the alular, major and minor digits. Os coxae comprised the ilium, ischium and pubis. Their mean lengths were 36.00 ± 0.82 cm, 32.00 ± 0.20 and 55.00 ± 0.2.9 cm, respectively. The femur was a stout short bone, that appeared shorter than the tibiotarsus. The mean length of the femur, tibiotarsus, and tarsometatarsus were 30.00 ± 0.23, 52.00 ± 0.50 and 46.00 ± 0.28 cm. Tibiotarsus was the longest bone in the pelvic limb. The fibula was a long bone (44.00 ± 0.41 cm) lying along the lateral surface of the tibiotarsus. The tarsometatarsus was a strong long bone formed by the fusion of the metatarsal (II, III, IV) and the distal row of tarsal bones. It was worth mentioning that metatarsal II was externally absent in adults. CONCLUSIONS: In the appendicular skeleton of ostrich, there were special characteristic features that were detected in our study; the clavicle was absent, the coracoid bone was composed of a sternal wing and scapular wing, the ulna was slightly longer in length than the radius. The coupled patellae i.e., the proximal and distal patella were observed; and the ostrich pedal digits were only two; viz., the third (III) and fourth (IV) digits.

CRISPR/Cas9-mediated knock-in of masu salmon (Oncorhyncus masou) elongase gene in the melanocortin-4 (mc4r) coding region of channel catfish (Ictalurus punctatus) genome.

Channel catfish, Ictalurus punctatus, have limited ability to synthesize Ω-3 fatty acids. The ccßA-msElovl2 transgene containing masu salmon, Oncorhynchus masou, elongase gene driven by the common carp, Cyprinus carpio, ß-actin promoter was inserted into the channel catfish melanocortin-4 receptor (mc4r) gene site using the two-hit two-oligo with plasmid (2H2OP) method. The best performing sgRNA resulted in a knockout mutation rate of 92%, a knock-in rate of 54% and a simultaneous knockout/knock-in rate of 49%. Fish containing both the ccßA-msElovl2 transgene knock-in and mc4r knockout (Elovl2) were 41.8% larger than controls at 6 months post-hatch (p = 0.005). Mean eicosapentaenoic acid (EPA, C20:5n-3) levels in Elov2 mutants and mc4r knockout mutants (MC4R) were 121.6% and 94.1% higher than in controls, respectively (p = 0.045; p = 0.025). Observed mean docosahexaenoic acid (DHA, C22:6n-3) and total EPA + DHA content was 32.8% and 45.1% higher, respectively, in Elovl2 transgenic channel catfish than controls (p = 0.368; p = 0.025). To our knowledge this is the first example of genome engineering to simultaneously target transgenesis and knock-out a gene in a commercially important aquaculture species for multiple improved performance traits. With a high transgene integration rate, improved growth, and higher omega-3 fatty acid content, the use of Elovl2 transgenic channel catfish appears beneficial for application on commercial farms.

Modeling aquifer storage and recovery in the eastern district of the United Arab Emirates using MODFLOW.

The Emirate of Abu Dhabi has relied on groundwater as a source of fresh water for several decades, which has resulted in the deterioration of non-renewable groundwater aquifers. This has led to the installation of desalination plants for fresh water supply. This research aims to increase strategic water reserves in the eastern district of Abu Dhabi by analyzing the best locations for aquifer storage and recovery (ASR). The ASR technology offers an opportunity to store large volumes of water for later beneficial use. This study explores an option of using excess desalination water for ASR recharge in the eastern district Al Ain region of Abu Dhabi. A limiting factor in the application of the ASR technology is the lack of suitable sites. Detailed hydrogeological and operational knowledge of the studied areas helped in identifying potential sites for ASR based on a scoring system. Determining best locations for managed aquifer recharge is a crucial design step. Five scenarios were studied at Al-Khrair and Al-Shuwaib sites in Al Ain region. Results show that a wider distribution of injection wells with intervals more than 1200 m is more suitable to overcome the excessive head buildup. Based on the adopted criteria, Al-Khrair was the best site for recharge followed by Al-Shuwaib. Al-Khrair site can be recharged at 64,000 m3 d-1 for seven years, while Al-Shuwaib site can be recharged at 64,000 m3 d-1 for only two years.

Improved Growth and High Inheritance of Melanocortin-4 Receptor (mc4r) Mutation in CRISPR/Cas-9 Gene-Edited Channel Catfish, Ictalurus punctatus.

Effects of CRISPR/Cas9 knockout of the melanocortin-4 receptor (mc4r) gene in channel catfish, Ictalurus punctatus, were investigated. Three sgRNAs targeting the channel catfish mc4r gene in conjunction with Cas9 protein were microinjected in embryos and mutation rate, inheritance, and growth were studied. Efficient mutagenesis was achieved as demonstrated by PCR, Surveyor® assay, and DNA sequencing. An overall mutation rate of 33% and 33% homozygosity/bi-allelism was achieved in 2017. Approximately 71% of progeny inherited the mutation. Growth was generally higher in MC4R mutants than controls (CNTRL) at all life stages and in both pond and tank environments. There was a positive relationship between zygosity and growth, with F1 homozygous/bi-allelic mutants reaching market size 30% faster than F1 heterozygotes in earthen ponds (p = 0.022). At the stocker stage (~ 50 g), MC4R × MC4R mutants generated in 2019 were 40% larger than the mean of combined CNTRL × CNTRL families (p = 0.005) and 54% larger than F1 MC4R × CNTRL mutants (p = 0.001) indicating mutation may be recessive. With a high mutation rate and inheritance of the mutation as well as improved growth, the use of gene-edited MC4R channel catfish appears to be beneficial for application on commercial farms.

Characteristics of Ultrasound and Magnetic Resonance Imaging of Normal Testes and Epididymis Besides Angiography of Testicular Artery in Dromedary Camel.

Decreasing male fertility encouraged the investigators to innovate accurate diagnostic non-invasive methods for detection of changes in the testicular parenchyma. Ultrasonography (US) has the potential to be used in this manner for decades, but magnetic resonance imaging (MRI) is still of limited application in animals for this purpose. The current study was designed to describe appearances and quantitative MRI attributes of the normal testes, epididymis besides angiography of testicular artery in camels. About 30 apparently healthy male dromedary camels aged 8-14 years were slaughtered during the rutting season. Immediately after slaughtering, the male gonads (n = 30 pairs of testicles and epididymis) were subjected to morphometric evaluation using a Vernier caliper and ultrasound scanning. Epididymial sperms were evaluated for motility, vitality and abnormality. MRI was performed for testes (n=16) by using a 1.5T Excite-II MRI apparatus of Sigma. Radiography and angioarchitecture of testicular artery (n=24) were done. Camel testicular length, width, and depth showed non-significant differences between a Vernier caliper or sonar. The MRI results revealed that both the testis and epididymis have homogenously intermediate signal (T1) and testes have hyperintense signal, with slightly lower signal in the epididymis (T2). In conclusion, both the ultrasonography and MRI techniques, with each respective computer-assisted imaging, could be used to detect the histomorphological changes of the camels' testicles. However, US imaging remains the first diagnostic technique for evaluating the reproductive health in men for its lower cost and accuracy. MRI is beneficial when the sonograms are inconclusive and/or equivocal. It shows the examined tissues in greater anatomical details compared to ultrasonography. Further studies are needed to compare between characteristics of US and MRI of normal testes and epididymis with testicular artery angiography in living camel during rut season and non-rut season and between normal healthy and affected diseased genitalia.

Effects of pomegranate peel and olive pomace supplementation on reproduction and oxidative status of rabbit doe.

The current study aimed to investigate the effects of pomegranate peel and olive pomace supplementation on the reproductive hormones, antioxidative status, reproductive capacity and maternal behaviour of rabbit does. Forty does were used for the experiments. The animals were randomly assigned to four groups of ten does. One group was fed on the control diet and was considered the control group (C). The second group was supplemented with 4.5% pomegranate peel in their diet (P), the third group was supplemented with 10% olive pomace in their diet (O), and the fourth group was supplemented with a mixture of pomegranate peel and olive pomace (PO). Compared with the control does, group P showed significantly increased serum levels of gonadotropic hormones and oestradiol-17ß two hours after mating, on the 20th day of lactation and after weaning; significantly increased progesterone levels at mid-pregnancy; and significantly increased in prolactin levels on the 10th day of lactation. Additionally, the results revealed significant increases in total DNA, protein concentration, litter size, milk yield and nest traits of groups P and PO. In conclusion, pomegranate peel supplementation improves the reproductive performance of does and increases their antioxidant parameters.

Direct and pleiotropic effects of the Masou Salmon Delta-5 Desaturase transgene in F1 channel catfish (Ictalurus punctatus).

Channel catfish (Ictalurus punctatus) is the primary culture species in the US along with its hybrid made with male blue catfish, I. furcatus. In an effort to improve the nutritional value of channel catfish, the masou salmon Δ5-desaturase like gene (D5D) driven by the common carp beta-actin promoter (ßactin) was inserted into channel catfish. The objectives of this study were to determine the effectiveness of ßactin-D5D for improving n-3 fatty acid production in F1 transgenic channel catfish, as well as examine pleiotropic effects on growth, proximate analysis, disease resistance, and other performance traits. Transgenic F1 channel catfish showed a 33% increase in the relative proportion of n-3 fatty acids coupled with a 15% decrease in n-6 fatty acids and a 17% decrease in n-9 fatty acids when compared to non-transgenic full-siblings (P < 0.01, P < 0.01, P < 0.01). However, while the relative proportion of n-3 fatty acids was achieved, the total amount of fatty acids in the transgenic fish decreased resulting in a reduction of all fatty acids. Insertion of the ßactin-D5D transgene into channel catfish also had large effects on the body composition, and growth of channel catfish. Transgenic channel catfish grew faster, were more disease resistant, had higher protein and moisture percentage, but lower fat percentage than full-sib controls. There were sex effects as performance changes were more dramatic and significant in males. The ßactin-D5D transgenic channel catfish were also more uniform in their fatty acid composition, growth and other traits.

Embryonic thermal manipulation of Japanese quail: effects on embryonic development, hatchability, and post-hatch performance.

Embryonic thermal manipulation led to several modifications in molecular, physiological, and biochemical parameters which affect pre- and post-hatch growth performance. The current study aims to elucidate the onset and long-term effects of intermittent thermal manipulations (TM) during two-time windows, early/late, of embryogenesis in Japanese quail (Coturnix japonica) on embryonic development, hatchability, muscle histogenesis, and post-hatch growth performance. Four groups were created; quail eggs in the control group were incubated at 37.7 °C and relative humidity (RH) 55%. Three thermally treated groups were incubated intermittently at 41 °C and 65% RH intermittently (3 h/day): early embryogenesis group (EE) was thermally treated during embryonic days (ED) 6-8, late embryogenesis group (LE) was thermally treated during (ED12-ED14), and early and late embryogenesis group (EL) was thermally manipulated in both time windows. Relative embryo weights in EL and EE were significantly lighter than those in LE and Ctrl groups. The hatched chicks were reared under optimal managemental conditions (three replicates per treatment). Average daily feed intake was recorded, and feed conversion ratio (FCR) was calculated. Histological and quantitative analyses of muscle fibers were performed. The results revealed that TM led to significant hypertrophy of quail breast muscle in (EE). Intermittent short-term (3-6 h) thermal manipulation (39-40 °C) protocols during early embryogenesis (ED6-ED8) could be recommended to enhance muscle mass growth and breast muscle yield in the Japanese quail.

Silver Nanoparticles as Modulators of Myogenesis-Related Gene Expression in Chicken Embryos.

The present study was conducted to investigate the effects of colloidal nanoparticles of silver (Nano-Ag) on the expression of myogenesis-related genes in chicken embryos. The investigated genes included the members of the myogenic regulatory factors family (MRFs) and myocyte enhancer factor 2A (MEF2A) genes. A total of 200 fertilized broiler eggs (Indian River) were randomly distributed into four groups; non-injected control, injected control with placebo, treatment I in ovo injected with 20 ppm Nano-Ag, and treatment II in ovo injected with 40 ppm Nano-Ag. The eggs were then incubated for 21 days at the optimum temperature and humidity conditions. Breast muscle tissues were collected at the 5th, 8th, and 18th days of the incubation period. The mRNA expression of myogenic determination factor 1 (MYOD1), myogenic factor 5 (MYF5), myogenic factor 6 (MYF6), myogenin (MYOG), and MEF2A was measured at the three sampling points using real-time quantitative PCR, while MYOD1 protein expression was evaluated on day 18 using western blot. Breast muscle tissues were histologically examined on day 18 to detect the changes at the cellular level. Our results indicate that myogenesis was enhanced with the low concentration (20 ppm) of Nano-Ag due to the higher expression of MYOD1, MYF5, and MYF6 at the transcriptional level and MYOD1 at the translational level. Moreover, histological analysis revealed the presence of hyperplasia (31.4% more muscle fibers) in treatment I (injected with 20 ppm). Our findings indicate that in ovo injection of 20 ppm Nano-Ag enhances the development of muscles in chicken embryos compared with the 40-ppm dosage and provide crucial information for the use of silver nanoparticles in poultry production.

Relationship between angioarchitecture of the testicular artery and spermiogram parameters in Egyptian buffalo bulls (bubalus bubalis).

The current study aimed to investigate the effect of testicular artery angioarchitecture on the spermiogram parameters in Egyptian Buffalo bulls. Eight adult buffalo bulls aged between 2 and 8 years were used for semen evaluation. For anatomical studies, the masculine gonads were collected after slaughtering 30 adult bulls and prepared for injection by different masses (Urographine® , Latex and Epoxy) through the testicular artery. The mass activity of the ejaculate was assessed immediately after collection. The sperm motility in fresh bull ejaculate was more than 80%. The overall mean percentage of sperm abnormalities was <18%. The recorded sperm abnormalities were mostly secondary one including distal protoplasmic droplet, fragmented tail, detached head, detached galea capitis and bent tail. The highest percentage of sperm viability was recorded just after sperm collection (alive > 85%). The results revealed that testicular artery can be divided into three parts (abdominal, funicular and marginal parts) along its course. The coils of the funicular part forming a cone-like structure with its base fixed to the head of the testis. Two epididymal branches to the head and tail of epididymis emanate from the funicular part which continues as pars marginalis on the lateral surface of testis before its division into the lateral and medial testicular arteries on approaching the tail extremity of the testis. The increase in the length of the testicular artery with increase in the size of the testes played a great role in the degree of complexity of the architectural vascular patterns. The degree of complexity is affected by the number of coils formed by the vessel. The increase in the convolutions of the vessel will reduce the speed of blood flow to the gonads. Thus in turn will enabling the thermoregulatory mechanism to work more efficiently and will affect the semen value.

The Y chromosome sequence of the channel catfish suggests novel sex determination mechanisms in teleost fish.

BACKGROUND: Sex determination mechanisms in teleost fish broadly differ from mammals and birds, with sex chromosomes that are far less differentiated and recombination often occurring along the length of the X and Y chromosomes, posing major challenges for the identification of specific sex determination genes. Here, we take an innovative approach of comparative genome analysis of the genomic sequences of the X chromosome and newly sequenced Y chromosome in the channel catfish. RESULTS: Using a YY channel catfish as the sequencing template, we generated, assembled, and annotated the Y genome sequence of channel catfish. The genome sequence assembly had a contig N50 size of 2.7 Mb and a scaffold N50 size of 26.7 Mb. Genetic linkage and GWAS analyses placed the sex determination locus within a genetic distance less than 0.5 cM and physical distance of 8.9 Mb. However, comparison of the channel catfish X and Y chromosome sequences showed no sex-specific genes. Instead, comparative RNA-Seq analysis between females and males revealed exclusive sex-specific expression of an isoform of the breast cancer anti-resistance 1 (BCAR1) gene in the male during early sex differentiation. Experimental knockout of BCAR1 gene converted genetic males (XY) to phenotypic females, suggesting BCAR1 as a putative sex determination gene. CONCLUSIONS: We present the first Y chromosome sequence among teleost fish, and one of the few whole Y chromosome sequences among vertebrate species. Comparative analyses suggest that sex-specific isoform expression through alternative splicing may underlie sex determination processes in the channel catfish, and we identify BCAR1 as a potential sex determination gene.

Effects of CRISPR/Cas9 dosage on TICAM1 and RBL gene mutation rate, embryonic development, hatchability and fry survival in channel catfish.

The current study was conducted to assess the effects of microinjection of different dosages of guide RNA (gRNA)/Cas9 protein on the mutation rate, embryo survival, embryonic development, hatchability and early fry survival in channel catfish, Ictalurus punctatus. Guide RNAs targeting two of the channel catfish immune-related genes, toll/interleukin 1 receptor domain-containing adapter molecule (TICAM 1) and rhamnose binding lectin (RBL) genes, were designed and prepared. Three dosages of gRNA/Cas9 protein (low, 2.5 ng gRNA/7.5 ng Cas9, medium, 5 ng gRNA/15 ng Cas9 and high, 7.5 ng gRNA/22.5 ng Cas9) were microinjected into the yolk of one-cell embryos. Mutation rate increased with higher dosages (p < 0.05). Higher dosages increased the mutation frequency in individual embryos where biallelic mutations were detected. For both genes, microinjection procedures increased the embryo mortality (p < 0.05). Increasing the dosage of gRNA/Cas9 protein increased the embryo mortality and reduced the hatching percent (p < 0.05). Embryonic development was delayed when gRNAs targeting RBL gene were injected. Means of fry survival time were similar for different dosages (p > 0.05). The current results lay the foundations for designing gene editing experiments in channel catfish and can be used as a guide for other fish species.

Microinjection of CRISPR/Cas9 Protein into Channel Catfish, Ictalurus punctatus, Embryos for Gene Editing.

The complete genome of the channel catfish, Ictalurus punctatus, has been sequenced, leading to greater opportunities for studying channel catfish gene function. Gene knockout has been used to study these gene functions in vivo. The clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 (CRISPR/Cas9) system is a powerful tool used to edit genomic DNA sequences to alter gene function. While the traditional approach has been to introduce CRISPR/Cas9 mRNA into the single cell embryos through microinjection, this can be a slow and inefficient process in catfish. Here, a detailed protocol for microinjection of channel catfish embryos with CRISPR/Cas9 protein is described. Briefly, eggs and sperm were collected and then artificial fertilization performed. Fertilized eggs were transferred to a Petri dish containing Holtfreter's solution. Injection volume was calibrated and then guide RNAs/Cas9 targeting the toll/interleukin 1 receptor domain-containing adapter molecule (TICAM 1) gene and rhamnose binding lectin (RBL) gene were microinjected into the yolk of one-cell embryos. The gene knockout was successful as indels were confirmed by DNA sequencing. The predicted protein sequence alterations due to these mutations included frameshift and truncated protein due to premature stop codons.

Generation of Myostatin Gene-Edited Channel Catfish (Ictalurus punctatus) via Zygote Injection of CRISPR/Cas9 System.

The myostatin (MSTN) gene is important because of its role in regulation of skeletal muscle growth in all vertebrates. In this study, CRISPR/Cas9 was utilized to successfully target the channel catfish, Ictalurus punctatus, muscle suppressor gene MSTN. CRISPR/Cas9 induced high rates (88-100%) of mutagenesis in the target protein-encoding sites of MSTN. MSTN-edited fry had more muscle cells (p < 0.001) than controls, and the mean body weight of gene-edited fry increased by 29.7%. The nucleic acid alignment of the mutated sequences against the wild-type sequence revealed multiple insertions and deletions. These results demonstrate that CRISPR/Cas9 is a highly efficient tool for editing the channel catfish genome, and opens ways for facilitating channel catfish genetic enhancement and functional genomics. This approach may produce growth-enhanced channel catfish and increase productivity.

Aquaculture genomics, genetics and breeding in the United States: current status, challenges, and priorities for future research.

Advancing the production efficiency and profitability of aquaculture is dependent upon the ability to utilize a diverse array of genetic resources. The ultimate goals of aquaculture genomics, genetics and breeding research are to enhance aquaculture production efficiency, sustainability, product quality, and profitability in support of the commercial sector and for the benefit of consumers. In order to achieve these goals, it is important to understand the genomic structure and organization of aquaculture species, and their genomic and phenomic variations, as well as the genetic basis of traits and their interrelationships. In addition, it is also important to understand the mechanisms of regulation and evolutionary conservation at the levels of genome, transcriptome, proteome, epigenome, and systems biology. With genomic information and information between the genomes and phenomes, technologies for marker/causal mutation-assisted selection, genome selection, and genome editing can be developed for applications in aquaculture. A set of genomic tools and resources must be made available including reference genome sequences and their annotations (including coding and non-coding regulatory elements), genome-wide polymorphic markers, efficient genotyping platforms, high-density and high-resolution linkage maps, and transcriptome resources including non-coding transcripts. Genomic and genetic control of important performance and production traits, such as disease resistance, feed conversion efficiency, growth rate, processing yield, behaviour, reproductive characteristics, and tolerance to environmental stressors like low dissolved oxygen, high or low water temperature and salinity, must be understood. QTL need to be identified, validated across strains, lines and populations, and their mechanisms of control understood. Causal gene(s) need to be identified. Genetic and epigenetic regulation of important aquaculture traits need to be determined, and technologies for marker-assisted selection, causal gene/mutation-assisted selection, genome selection, and genome editing using CRISPR and other technologies must be developed, demonstrated with applicability, and application to aquaculture industries.Major progress has been made in aquaculture genomics for dozens of fish and shellfish species including the development of genetic linkage maps, physical maps, microarrays, single nucleotide polymorphism (SNP) arrays, transcriptome databases and various stages of genome reference sequences. This paper provides a general review of the current status, challenges and future research needs of aquaculture genomics, genetics, and breeding, with a focus on major aquaculture species in the United States: catfish, rainbow trout, Atlantic salmon, tilapia, striped bass, oysters, and shrimp. While the overall research priorities and the practical goals are similar across various aquaculture species, the current status in each species should dictate the next priority areas within the species. This paper is an output of the USDA Workshop for Aquaculture Genomics, Genetics, and Breeding held in late March 2016 in Auburn, Alabama, with participants from all parts of the United States.

Drag reduction using lubricant-impregnated surfaces in viscous laminar flow.

Lubricant-impregnated surfaces (LIS), where micro/nanotextured surfaces are impregnated with lubricating liquids, have received significant attention for their robust, superslippery properties. In this study, we systematically demonstrate the potential for LIS to reduce drag in laminar flows. We present a scaling model that incorporates the viscosity of the lubricant and elucidates the dependence of drag reduction on the ratio of the viscosity of the working fluid to that of the lubricant. We experimentally validate this dependence in a cone and plate rheometer and demonstrate a drag reduction of 16% and slip length of 18 µm in the case where the ratio of working fluid viscosity to lubricant viscosity is 260.

Binary colloidal structures assembled through Ising interactions.

New methods for inducing microscopic particles to assemble into useful macroscopic structures could open pathways for fabricating complex materials that cannot be produced by lithographic methods. Here we demonstrate a colloidal assembly technique that uses two parameters to tune the assembly of over 20 different pre-programmed structures, including kagome, honeycomb and square lattices, as well as various chain and ring configurations. We programme the assembled structures by controlling the relative concentrations and interaction strengths between spherical magnetic and non-magnetic beads, which behave as paramagnetic or diamagnetic dipoles when immersed in a ferrofluid. A comparison of our experimental observations with potential energy calculations suggests that the lowest energy configuration within binary mixtures is determined entirely by the relative dipole strengths and their relative concentrations.