Journal articles: 'Pig-male reproductor' – Grafiati (2024)

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Abstract. The sperm flagella 2 (SPEF2) gene is essential for normal sperm tail development and male fertility. To fully characterize the structure of the mutation and to further study the function of the pig SPEF2 gene, we explored the insertion/deletion (indel) and novel single-nucleotide polymorphisms (SNPs) within the pig SPEF2 gene, and tested their associations with the testicular sizes in male Large White (LW) and Landrace (LD) pigs from China. Herein, a large insertion located at the SPEF2 gene in chromosome 16 was found, and two alleles of "I" (insertion) and "D" (deletion) were designated. Allele "D" was dominant in all analyzed pigs. Two novel SNPs (namely (NC_010458) g.19642G > A, resulting in AfaI aCRS PCR–PFLP, and g.19886C > G, resulting in EcoRI aCRS PCR–PFLP) were found in LW and LD pigs. Association testing revealed that g.19886C > G was significantly associated with the testis long circumference (TLC) in LW pigs (P < 0.05), suggesting that this SNP would be the DNA marker for the marker-assisted selection (MAS) in reproduction traits. This preliminary result indicates that the pig SPEF2 gene had significant effects on male reproduction traits. These findings could not only extend the spectrum of genetic variations in the pig SPEF2 gene but also contribute to implementing MAS in genetics and breeding in pigs.

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Bone morphogenetic protein 15 (BMP15) is a member of the transforming growth factor-β (TGFB) superfamily that plays an essential role in mammalian ovary development, oocyte maturation and litter size. However, little is known regarding the expression pattern and biological function of BMP15 in male gonads. In this study we established, for the first time, a transgenic pig model with BMP15 constitutively knocked down by short hairpin (sh) RNA. The transgenic boars were fertile, but sperm viability was decreased. Further analysis of the TGFB/SMAD pathway and markers of reproductive capacity, namely androgen receptor and protamine 2, failed to identify any differentially expressed genes. These results indicate that, in the pig, the biological function of BMP15 in the development of male gonads is not as crucial as in ovary development. However, the role of BMP15 in sperm viability requires further investigation. This study provides new insights into the role of BMP15 in male pig reproduction.

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ABSTRACTEXPEDIENCY OF INCREASING PIG POPULATION BY ARTIFICIAL INSEMINATION TECHNIQUE IN NORTH SULAWESI PROVINCE.The objective of this study was to evaluate dominant factors affecting pig population breeding by artificial insemination (AI) in North Sulawesi province. sem*n of boar (Landrace, Yorkshire and Duroc breeds) were usedto mate the same breeds using AI.Data of 345 piglets from 33 sows were involved in this study. Reproductive data of sows and productivity of piglet form AI were observed at laboratory and field belonging to farmers in Minahasa regency. Pair data of male and female effect were analyzed by t-test for variables.Dominant reproduction factors affecting pig population breeding in North Sulawesi were includingservice per conception, Farrowing rate, AI service efficiency, and Farrowing index.Group of male and superior piglets were higher in productivity that those of female and inferior piglets. Preweaning Average Daily Gain (PW-ADG)groups of inferior (84.85g/h/d)were significantly lower compared with normal (86.59g/h/d), butnot significantly different with PW-ADG of superior birth weight group (85.64 g/h/d).PW-ADG.Thelitter size produced from sem*n bya boar naturally mated with the same sowsreached the average of 21 piglets per year, while those by a boar mated using AI technique with the same sowsreached the average of 225 piglets per year. Keywords:Artificial insemination. Pig reproduction, North Sulawesi province

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In spite of the vast distribution and considerable impact on human and animal health of Ascaris suum and A. lumbricoides, little is known of the sexual biology and reproductive capacity of these intestinal nematodes. By oral transfer of adult female worms to previously parasite-naive pigs we show that in the absence of males the egg production ceases after 2–3 weeks. Such females readily resume egg production a few days after oral transfer of male worms. These observations throw light on an important aspect of Ascaris biology, but also pave the way for possible experimental interbreeding between the human and pig Ascaris species.

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Cypermethrin is a large spectrum action insecticide, globally employed to control pests in agriculture and some human and domestic animals ectoparasites. This study aimed to evaluate its toxicity and reproduction impairment in male guinea pig. Forty adult male guinea pigs were divided into 4 groups and orally submitted to 0, 92, 137.5 and 275 mg/kg body weight/day for 90 days. The weight of the liver increased significantly, while that of kidneys decreased significantly in treated animals compared to controls. Serum concentrations of creatinine, urea, ALAT, ASAT, total cholesterol, prostatic acid phosphatase increased significantly, while the testicular total protein level decreased significantly in groups given the insecticide relatively to the control. The testes weight, libido, serum level of testosterone, mobility, sperm count and the percentage of spermatozoa with entire plasma membrane decreased significantly in animals exposed to cypermethrin with reference to controls. The percentages of abnormal spermatozoa increased significantly in animals submitted to 137.5 or 275 mg/kg body weight (bw) of cypermethrin compared to control ones. On the testis histological sections of pesticide-treated animals, immature germinal cells were observed in the lumen of seminiferous tubules. Cypermethrin was toxic in male guinea pig and damaged reproductive parameters.

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The pursuit of enhanced prolificacy and fertility has been a major goal for the British pig industry for some years and throughout the 1960s to the 1990s advances were made that were generally due to better nutrition and management but genetics also played a significant role. Health status also influences reproduction and the production of faster growing pigs to slaughter. In recent years as farms have become larger and more intensive, our national health status may have been in decline to the detriment of female and male reproduction. Figures 13.1, 13.2 and 13.3 illustrate the changes we have seen in 3 important reproduction characteristics, using data from MLC yearbooks, 1970 to 2003. Farrowing index improved rapidly in the 1970s due, in part, to early weaning reducing the average farrowing interval, but also increased producer awareness of mating management and heat detection procedures helped to push this upwards.

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Abstract The effect of endogenous estrogens on the male reproductive tissues and development of gametes is an essential for the reproductive success. Estrogens affect the target cells via estrogen receptors (ERs) by both genomic and non-genomic pathways. The ER localization in the testis, epididymis, and sperm cells is a key to understanding the effect of estrogens on the sperm development, maturation, and function. The ER detection in male reproductive tissues and sperm cells at different development stages is described in representative mammalian species (human, mouse, rat, horse, and pig), in which the ER localization has been most described. According to various authors the ER occurrence in the male reproductive tissues and spermatozoa is quite distinct. Discrepancy in the published results is probably caused either by the application of different tissue preparation methods, or the choice of specific antibodies. Inconsistent findings should be subjected to further investigation to better understand the role of ERs in the male gamete development and mammalian reproduction

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Several studies have recently investigated the role of leptin, the adipocyte-secreted hormone, in the growth and reproduction of rodents, humans, and domestic animals. The present study was designed to explore the expression of leptin and its receptor in pig spermatozoa. Successful Western blot evidenced a 16 kDa band for leptin and six isoforms, ranging from 120 to 40 kDa, for the leptin receptor. Both leptin and leptin receptor were interestingly located at sperm acrosomal level, suggesting their involvement in the oocyte fertilization events. In fact, both capacitation indexes and acrosin activity were enhanced by leptin, and these effects were reduced by the anti-leptin receptor antibody. Afterwards, we investigated the main transduction pathways regulated by the hormone. Our results showed that, in pig sperm, leptin can trigger the signal transducer and activator of transcription 3, a classical component of cytokine signal transduction pathways, whose expression has not been previously reported in male gamete; in addition it was found constitutively activated. Besides, leptin was able to induce the activation of phosphatidylinositol phosphate kinase 3 and MAP kinase pathways as well as of BCL2, a known antiapoptotic protein. These data address to a role of leptin and its receptor on pig sperm survival. The presence of leptin and its receptor in pig sperm suggests that they, through an autocrine short loop, may induce signal transduction and molecular changes associated with sperm capacitation and survival.

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This study aimed to evaluate the allometric growth of the Araucan pig breed, a creole breed from Arauca, East Colombia, locally known as “Sabaneros”, in relation to different quantitative traits and considering genders separately. To do this, a total of 31 male and 27 female Araucan pigs, ranging from 4 to 48 months of age, were studied in order to evaluate their growth patterns, using a multivariate approach. Animals belonged to different farms (“fincas”) of the Department of Arauca, Colombia. From each individual, 10 quantitative traits were obtained: face width, croup height, croup length, croup width, tail base height, hock height, loin height, cannon length, and length and width of ear. Our results, which must be interpreted as preliminary, showed that the Araucan pig is allometrically monomorphic as sexual differences do not increase with body size. We suggest that although males and females have evidently different reproductive roles, during growth they shift the allocation of energy to structures linked to environmental adaptation rather than those linked to reproduction.

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Cannabinoids regulate biological processes governed by the hypothalamus including, but not limited to, energy homeostasis and reproduction. The present study sought to determine whether cannabinoids modulate A-type K+ currents ( IA) in neurons of the hypothalamic arcuate nucleus (ARC). Whole cell patch-clamp recordings were performed in slices through the ARC prepared from castrated female and male guinea pigs. Forty percent of guinea pig ARC neurons exhibited a transient outward current that was antagonized by high (mM) concentrations of 4-aminopyridine and (100 nM) rHeteropodatoxin-2. Five of these neurons also were immunopositive for both β-endorphin and the Kv4.2 channel subunit. Bath application of the CB1 receptor agonists WIN 55,212–2 (1 μM) or ACEA (1 μM) selectively induced a rightward shift in the inactivation curve for the IA, significantly increasing the half-maximal voltage without affecting the peak current magnitude, in neurons from female but not male animals. The CB1 receptor antagonist AM251 (1 μM) reversed this action. Collectively, these data reveal that guinea pig ARC neurons, including proopiomelanocortin neurons, express a prominent IA that is positively modulated by cannabinoids in a sex-specific way by altering the voltage dependence of its inactivation. The resultant inhibitory effect on this neuronal population may shed some insight into the mechanism(s) by which cannabinoids influence hypothalamic function.

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Abstract Spermatogonial stem cells (SSCs) self-renew and contribute genetic information to the next generation. Pig is wildly used as a model animal for understanding reproduction mechanisms of human being. Inducing directional differentiation of porcine SSCs may be an important strategy in exploring the mechanisms of spermatogenesis and developing better treatment methods for male infertility. Here, we established an in-vitro culture model for porcine small seminiferous tubule segments, to induce SSCs to differentiate into single-tail haploid spermatozoa. The culture model subsequently enabled spermatozoa to express the sperm-specific protein acrosin and oocytes to develop to blastocyst stage after round spermatid injection. The addition of retinoic acid (RA) to the differentiation media promoted the efficiency of haploid differentiation. RT-PCR analysis indicated that RA stimulated the expression of Stra8 but reduced the expression of NANOS2 in spermatogonia. Genes involved in post-meiotic development, transition protein 1 (Tnp1) and protamine 1 (Prm1) were upregulated in the presence of RA. The addition of an RA receptor (RAR) inhibitor, BMS439, showed that RA enhanced the expression of cAMP responsive-element binding protein through RAR and promoted the formation of round spermatids. We established an efficient culture system for in-vitro differentiation of pig SSCs. Our study represents a model for human testis disease and toxicology screening. Molecular regulators of SSC differentiation revealed in this study might provide a therapeutic strategy for male infertility.

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The Meishan pig is a traditional Chinese native breed, known for its excellent reproduction performance that is widely used in commercial pig production through two-way or three-way crossbreeding systems. However, the lean meat yield of Meishan crossbred pigs is still very low and cannot meet the market demand. To evaluate the lean meat yield of Meishan crossbred pigs, six wild-type Meishan sows were artificially inseminated by using the MSTN+/− Duroc boar sem*n in this experiment. Some reproductive performance-related traits of Meishan sows were recorded to ensure that sem*n from MSTN knockout Duroc boar did not affect offspring production, including total births, live births, sex, and litter weight. In total, 73 piglets were obtained and 63 were alive. Male to female ratio was close to 1: 1. because of factors such as disease, only 43 pigs were utilized, including 28 MSTN mutant pigs (MSTN+/−) and 15 MSTN hom*ozygous pigs (MSTN+/+). We compared the growth performance and carcass performance of these full or half-sib populations and found that there were no differences between MSTN+/− and MSTN+/+ genotypes for live animal measures including average daily gain (ADG), body dimensions, or ultrasonic measurement of fat thickness when pigs were harvested after 120 days of feeding. Conversely, the MSTN+/− pigs had higher dressing percentage and lean meat percentage, lower level of carcass fat, larger longissimus muscle area, less percentage of skin and skeleton, thinner average backfat thickness, and lower intramuscular fat (IMF) content than MSTN+/+ pigs. In conclusion, the production of MSTN+/− mutant progeny from Meishan females resulted in improved carcass composition, providing a feasible solution to improve the lean meat yield of Chinese local fat-type pig breeds.

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ABSTRACT Peroxisome proliferator-activated receptor gamma (PPARγ) acts as a ligand activated transcription factor and regulates processes, such as energy homeostasis, cell proliferation and differentiation. PPARγ binds to DNA as a heterodimer with retinoid X receptor and it is activated by polyunsaturated fatty acids and fatty acid derivatives, such as prostaglandins. In addition, the insulin-sensitizing thiazolidinediones, such as rosiglitazone, are potent and specific activators of PPARγ. PPARγ is present along the hypothalamic–pituitary–testis axis and in the testis, where low levels in Leydig cells and higher levels in Sertoli cells as well as in germ cells have been found. High amounts of PPARγ were reported in the normal epididymis and in the prostate, but the receptor was almost undetectable in the seminal vesicles. Interestingly, in the human and in pig, PPARγ protein is highly expressed in ejacul*ted spermatozoa, suggesting a possible role of PPARγ signaling in the regulation of sperm biology. This implies that both natural and synthetic PPARγ ligands may act directly on sperm improving its performance. Given the close link between energy balance and reproduction, activation of PPARγ may have promising metabolic implications in male reproductive functions. In this review, we first describe PPARγ expression in different compartments of the male reproductive axis. Subsequently, we discuss the role of PPARγ in both physiological and several pathological conditions related to the male fertility.

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Stimulated by retinoic acid 8 (STRA8) is a protein that is required for meiotic initiation in both male and female gametes in vertebrates. It is also expressed in embryonic germ cells and neonatal male germ cells of mice. The utility of using the Stra8 promoter to recognise and isolate pre-meiotic male germ cells has been reported by others in the mouse. In order to mark germ cells in male pigs, we cloned 1.6 kb of the mouse Stra8 promoter and used it to develop a reporter plasmid using mitochondrial-localised enhanced yellow fluorescent protein (mEYFP). The Stra8-mEYFP transgenic male pigs were produced using somatic cell nuclear transfer. The mEYFP reporter was expressed and easily detectable in the live germ cells of the mature animals and could be observed during tissue culture. The mitochondrial-localised expression of the EYFP reporter was helpful in observing the size and stage of the germ cell. The mEYPF protein was found to be expressed only in the testis of the transgenic pigs using Western blot analysis, whereas endogenous STRA8 protein was also detected in the lung and brain. Fluorescent immunohistochemistry of testicular sections of the transgenic pigs indicated a similar expression pattern to that of the endogenous STRA8 protein. There was an overlap in the expression of the mEYFP and the endogenous STRA8 protein; however, it was observed that the mEYFP protein was present at an earlier stage of spermatogenesis than the STRA8 protein. Immunocytochemistry performed on plated tubules similarly showed varying intensity in expression between the mEYFP transgene and the endogenous STRA8. The difference in the timing of protein expression may be due to the model created or the use of the mouse Stra8 promoter for the expression of mEYFP. Alternatively, the lag in expression between that of the endogenous STRA8 and mEYFP protein may be due to attenuated translation of the Stra8 mRNA. This transgenic model should be useful for the study of reproduction, development, transplantation, biotechnology, and culture of the pig male germ line. Supported by North Carolina Agricultural Research Service 02234.

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Abstract. Transmembrane protein 95 (TMEM95) is located on the acrosomal membrane of the sperm head involved in the acrosome reaction; thus, it is regarded as affecting spermatogenesis and reproduction traits. The aim of this study was to explore the novel single nucleotide polymorphisms (SNPs) within the pig TMEM95 gene as well as to evaluate their associations with the testicular sizes in male Landrace (LD) and Large White (LW) breeds. After pool sequencing and bioinformatics analysis, only one novel coding SNP was found in exon 1, namely NC_010454.3: g.341T > C, resulting in a synonymous mutation (A47A). This SNP could be genotyped using the StuI polymerase chain reaction–restriction fragment length polymorphism (PCR-RFLP) assay. The minor allelic frequencies (MAFs) were 0.259 and 0.480 in the LD and LW breeds. Their polymorphism information content (PIC) values were 0.310 and 0.375. The LW population was at the Hardy–Weinberg equilibrium (HWE) (p > 0.05), whereas the LD population was not (p < 0.05). Association analyses demonstrated that a significant relationship was found between this A47A polymorphism and testis weight at 40 days of age in the LW population (p = 0.047), and the heterozygote individuals showed lower testis weight than those with other genotypes. Moreover, this SNP was significantly associated with three testis measurement traits at 15 days of age in the LW population (p < 0.05); the individuals with genotypes TT and TC showed consistently superior testis measurement traits than those with genotype CC. These findings demonstrate that the A47A polymorphism had a significant effect on testis measurement traits, suggesting that the TMEM95 gene could be a candidate gene associated with reproductive traits. These results could contribute to breeding and genetics programs in the pig industry via DNA marker-assisted selection (MAS).

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In order to achieve fertilization sperm cells first need to successfully interact with the zona pellucida. Before reaching the zona pellucida the sperm cell undergoes extensive remodeling both in the male and female genital tract. These changes serve to mediate optimal recognition of the zona pellucida in the oviduct (primary zona pellucida binding). Optimal sperm-zona interactions are crucial for porcine oocyte fertilization: The zona pellucida- attached sperm cell is triggered to undergo the acrosome reaction and will also become hypermotile. Together these two responses allow the sperm cell to drill through the zona pellucida (secondary zona pellucida binding) this coincides with local sperm zona drilling so that a few sperm can reach the perivitellin space. This delaying strategy allows only one sperm cell in a given time-point to bind and fuse with the oocyte (fertilization) and thus minimizes the risk of polyspermy. The polyspermy block is essentially executed by the fertilized oocyte that immediately secretes its cortical granule content into the perivitellin space. This content blocks sperm-oocyte fusion either by sticking to the oolemma or by the induction of a biochemical reaction of the zona pellucida (zona pellucida hardening). The cortical reaction thus blocks sperm-zona pellucida binding and/or sperm-zona pellucida drilling. Note that zona pellcuida interactions under pig IVF conditions relatively frequently result in polyspermy. It is not clear whether this is also the case in vivo after natural mating. More importantly we do not know how other artificial reproductive technologies affect polyspermy rates. This is especially relevant for new sperm treatments and insemination technologies in which sperm are activated by capacitation media essentially mimicking the IVF media. Therefore, better understanding of sperm activation and of the arrangement of proteins involved in zona pellucida interactions are relevant for designing strategies to further improve pig reproduction.

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This study was conducted to evaluate the effect of different maturation times on the output of intracytoplasmic sperm injection (ICSI) with pig oocytes precultured with roscovitine, a specific inhibitor of MPF kinase activity (Meijer and Raymond 2003 Acc. Chem. Res. 36, 417–425). Cumulus-oocyte complexes (COCs) were collected from nonatretic follicles from slaughtered gilts. Just after recovery, oocytes (n = 456) were cultured in the presence of 50 μ­M roscovitine for 28–30 h, which maintains them at the GV stage until use without affecting further development (Coy et al. 2004 Human Reprod. 19, 41–42). Then, oocytes were matured in a conventional maturation system (Coy et al. 2002 Reproduction 124, 279–288) for 36, 40, or 44 h. Matured oocytes were fertilized by ICSI. Injection was performed as described by Martin (2000 Biol. Reprod. 63, 109–112) but in D-PBS medium with 10% FCS. Injected oocytes were transferred to TALP medium (Rath et al. 1999 J. Anim. Sci. 77, 3346–3352) for 24 h, after which they were fixed and Hoechst-stained to check for fertilization. Results (data were analyzed by ANOVA and Tukey test) showed similar activation rates (oocytes with female pronucleus) for 36, 40, and 44 h (83.6%, 81.7%, and 90.7%, respectively). Differences were observed in male pronuclear formation (80.2%, 67.6%, and 57.8%) and in putative embryos (zygotes showing two pronuclei or two cell stage 24 hpi; 74.3%, 62.9%, and 55.1%) for 36, 40, and 44 h, respectively (P < 0.05), where the 36-h group showed the best result. Data show that pre-treatment of pig oocytes with roscovitine and further maturation for 36 h can improve the output of ICSI. This work was supported by Ministerio de Ciencia y Tecnología (AGL2003-03144).

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Testis tissue grafting presents an option for preservation of genetic material when sperm recovery is not possible. Grafting of testis tissue from sexually immature males to immunodeficient mice results in germ cell differentiation and production of fertilization-competent sperm from different mammalian species (Honaramooz et al. 2002 Nature 418, 778–781). However, the efficiency of testis tissue xenografting from adult donors has not been critically evaluated. Spermatogenesis was arrested at meiosis in grafts from mature horses (Rathi et al. 2006 Reproduction 131, 1091–1098) and hamsters (Schlatt et al. 2002 Reproduction 124, 339–346), and no germ cell differentiation occurred in xenografts of adult human testis tissue (Schlatt et al. 2006 Hum. Reprod. 21, 384–389). The objective of this study was to investigate survival and germ cell differentiation of testis xenografts from sexually mature donors of different species. Small fragments of testis tissue from 10 donor animals of 5 species were grafted under the back skin of immunodeficient, castrated male mice (n = 37, 2–6/donor). Donors were pig (8 months old), goat (18 months old and 4 years old) (n = 2), bull (3 years old), donkey (13 months old), and rhesus monkey (3, 6, 11, and 12 years old). At the time of grafting, donor tissue contained elongated spermatids, albeit to different degrees (&gt;75% of seminiferous tubules in testis tissue from pig, goat, bull, and 6–12-year-old monkeys, and 33 or 66% of tubules in tissue from donkey or 3-year-old monkey, respectively). Grafts were recovered &lt;12 weeks (n = 14 mice), 12–24 weeks (n = 16 mice), and &gt;24 weeks (n = 7 mice) after grafting and classified histologically as completely degenerated (no tubules found), degenerated tubules (only hyalinized seminiferous tubules observed), or according to the most advanced type of germ cell present. Grafts from pig, goat, bull, and 6–12-year-old monkeys contained &gt;60% degenerated tubules or were completely degenerated at all time points analyzed. In contrast, in grafts from the 3-year-old monkey, only 18% of tubules were degenerated, 14% contained Sertoli cells only, 64% contained meiotic, and 4% haploid germ cells at 24 weeks after grafting. Similarly, donkey testis grafts recovered 12–24 weeks after grafting contained &lt;2% degenerated tubules, 46% of tubules had Sertoli cells only, 45% contained meiotic, and 7% haploid germ cells. These results show that survival and differentiation of germ cells in testis grafts from sexually mature mammalian donors is poor. However, better graft survival and maintenance of spermatogenesis occurred in donor tissue from donkey and 3-year-old monkey that were less mature at the time of grafting. Therefore, species and age-related differences appear to exist with regard to germ cell survival and differentiation in xenografts from adult donors. This work was supported by USDA/CSREES 03-35203-13486, NIH/NCRR 5-R01-RR17359-05, the Spanish Ministry of Education, and Science (BES-2004-4112).

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This review was carried out with the purpose of describing the main reproductive management used in the rearing of pigs. Currently, swine breeding in Brazil has average production parameters of 9.5 piglets / calving with 1.9 calving / year, with a genetic potential to reach 13 piglets / calving with 2.6 calving / year. To obtain these zootechnical indexes in pig production, it is necessary to invest in reproductive technologies such as artificial insemination, adequate facilities and frequent training of the workforce. In this sense, to be successful in reproduction, it becomes necessary to know the anatomy and physiology of the male and female reproductive system. The good reproductive performance of the animals also depends on the criterion of selection of the boar and the matrices applied in the breeding. It is fundamental to know the estrous cycle, as well as, the behavior of the nut in estrus to obtain success in artificial insemination or natural mating. The gestation period, as well as childbirth, are critical breeding phases and should be well conducted with the aim of avoiding or reducing piglet mortality. Therefore, the reproductive stages of pigs should be well conducted with special attention to food and sanitary management. The knowledge of the reproductive management by the producer and / or technician responsible for the farm is of fundamental importance to achieve good productive indexes

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Abstract This study was aimed at investigating the use of intra-testicular calcium chloride (CaCl2) and 4-vinylcyclohexene 1,2-monoepoxide (VCM) injections as a side effect-free alternative method for the control of reproduction in guinea pigs. Fifty male guinea pigs were randomly assigned to five groups. In all groups, the chemical agents were injected into both testes in 1% lidocaine hydrochloride. While Groups I, II and III were administered with a single dose (0.25 mL) of sterile physiological saline, 15 mg/100 g CaCl2, and 240 mg/kg VCM, respectively, Group IV and V received a daily dose of 15 mg/100 g CaCl2, and 240 mg/kg VCM for 3 days, respectively. On day 90 post-administration, all animals were weighed and later decapitated under ether anaesthesia. Blood and tissue (testis, liver, hypophysis and adrenal gland) samples were taken. Sperm samples from the cauda epididymis were examined for spermatological parameters. Blood was used for hormone analyses and tissue samples were examined histopathologically (haematoxylin-eosin) and immunohistochemically (Tunel staining). The epididymal sperm count decreased in all treatment groups. Excluding 2 animals, Group V displayed azoospermia. When compared to the control group, Group V displayed the highest prolactin and lowest testosterone levels, and Group III showed the highest testosterone level. Histopathological examination revealed no intoxication finding. Chemical castration with VCM may be a good alternative to surgical castration as it enables mass sterilization without postoperative risks in guinea pig.

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Transgenic (Tg) pigs with organ/tissue-specific fluorescence expression provide invaluable research tools for many types of studies, such as organogenesis analysis, in vitro tissue generation from pluripotent cells, and progenitor/stem cell transplantation therapy. We aimed to develop a Tg pig characterised by pancreas- and liver-specific fluorescence expression. A 8.4 kb transgene construct expressing Venus (green fluorescence) under the control of the mouse Pdx1 (pancreatic duodenal homeobox-1) promoter and a BAC-derived construct (170 kb) consisting of the whole-length porcine albumin (Alb) promoter and humanized Kusabira-Orange (huKO, red fluorescence) was introduced into porcine in vitro-matured oocytes using the intracytoplasmic sperm injection (ICSI)-mediated gene transfer method. Injected embryos were transferred to the oviducts of oestrus-synchronized recipients after culture for 1 to 3 days. The transfer of 370 Pdx1-Venus embryos into 4 recipients produce 22 (5.9%) fetuses/piglets, and 9 (40.9%) Tg pigs exhibited pancreas-specific Venus expression. Two (1 male and 1 female) founder Pdx1-Venus-Tg pigs were mated with wild-type (WT) pigs and produced 32 offspring in 3 litters, of which 16 (50.0%) were transgenic. Pancreas-specific Venus expression was inherited in these Tg offspring. The transfer of 523 Alb-huKO embryos into 4 recipients resulted in 19 (3.6%) piglets including a Tg female, which showed liver-specific huKO fluorescent expression. Expression of huKO was detected by RT-PCR exclusively in liver, but not in 7 other organs/tissues examined, including heart, lung, stomach, small intestine, spleen, kidney and skin. This founder Tg female produced a total of 12 non-Tg and 5 Tg offspring (in 2 litters) after mating with a WT boar. Liver-specific huKO expression was inherited in these Tg offspring. Furthermore, the mating of a female Pdx1-Venus pig with an Alb-huKO boar yielded 7 non-Tg and 10 Tg pigs. Four of these Tg pigs carrying both of the transgenes exhibited both pancreas-specific Venus and liver-specific huKO expression in single individuals. Double-Tg pigs with pancreas-specific green fluorescence and liver-specific red fluorescence grew normally, and tests of their reproduction ability are currently underway. These data demonstrate that transgene introduction by ICSI-mediated gene transfer into in vitro-matured oocytes is a feasible option for generating pigs expressing fluorescent proteins in a tissue-specific manner.

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Feral pigs (Sus scrofa) are an invasive species widely distributed throughout North and South America, as well as Australia. Feral pigs frequently destroy farm fields, causing $1.5 billion in damage annually in the United States alone. These pigs are known carriers of over 30 diseases, thereby posing a threat to livestock and public health. Current control methods are ineffective due to high fecundity rates. An orally consumed contraceptive bait may be effective, but currently none exists for pigs. In rats, an orally consumed contraceptive that affects both male and females has shown considerable promise. This bait contains 2 active ingredients: triptolide, a diterpenoid diepoxide isolated from the Chinese medicinal plant Tripterygium wildfordii, and 4-vinylcyclohexene diepoxide (VCD), an industrial chemical. Triptolide prevents ovarian follicle maturation and disrupts spermatogenesis; VCD acts in the female to cause premature ovarian failure and, in one report, disrupt male mouse spermatogenesis. There are no studies reported for these active ingredients on boar spermatogenesis. A proprietary pig bait has been developed containing these 2 ingredients and was fed to commercial (com.) and Sinclair (sin.) boars (com. n = 3, sin. n = 3) for 15 consecutive days to evaluate changes in reproductive parameters. Boars were given 250 g of bait twice daily that contained 0.25 mg of triptolide and 1.55 mM of VCD. ejacul*tes were collected from boars before, during, and after the treatment period (Day 0, 7, 15, 30, 37, 45, and 60) and were evaluated for possible impacts on fertility. ejacul*tes were able to be collected from 4 of 6 boars (com. n = 2, sin. n = 2). Previous studies indicate triptolide acts during the differentiation phase of spermatogenesis, suggesting a delayed onset of perturbation allowing for comparison of sem*n collected early in the study (Day 0, 7, and 15) to post-treatment samples. Progressive motility (PM) was rated on a 1 to 5 scale. Plasma testosterone and testes volume were also monitored at these time points. Statistical analysis was performed using Kruskal–Wallis test. No significant differences were found in plasma testosterone, testes volume, or sperm concentration. No significant differences in viability, morphology, or PM were found at Day 0, 7, or 15. However, differences (P < 0.05) were observed in sem*n parameters at Day 37 and 45 in comparison to D0, 7, and 15. The percentage of sperm with normal morphology at treatment Day 37 (3.3%) and 45 (3.3%) was lower than at Day 0, 7, and 15 (72.3%, P < 0.05). Differences (P < 0.05) were also found in viability at Day 37 (13.3%) and 45 (8%) in comparison to Day 0, 7, and 15 (71.6%). Differences were also found in PM at Day 37 (0.33) and 45 (1) in comparison to Day 0, 7, and 15 (4.8). These results suggest an impact on spermatogenesis, specifically spermiogenesis, suggesting a decrease in fertility due to lower quality of spermatozoa. Further studies are needed to evaluate the bait’s impact on feral pig reproduction.

Abstract:

The Dunkin Hartley is the most common guinea pig strain used in biomedical research, particularly for studies of asthma, allergy, infectious disease, reproduction, and osteoarthritis. Minimally invasive blood tests, such as complete blood counts and serum biochemistry profiles, are often collected for diagnostics and laboratory analyses. However, reference intervals for these assays have not yet been well-documented in this strain. The purpose of this study was to establish reference intervals for hematologic and biochemical parameters of Dunkin Hartley guinea pigs and determine age- and sex-related differences. Hematologic and biochemical parameters were retrospectively obtained from 145 male and 68 female guinea pigs between 2 and 15 months of age. All blood parameters were analyzed by a veterinary clinical pathology laboratory. Reference intervals were established according to the American Society for Veterinary Clinical Pathology guidelines. Age- and sex-related differences were determined using unpaired t-tests or nonparametric Mann-Whitney tests. Hematocrit, red blood cell distribution width, mean platelet volume, white blood cell count, heterophils, monocytes, eosinophils, glucose, blood urea nitrogen, creatinine, calcium, magnesium, total protein, albumin, globulin, cholesterol, aspartate aminotransferase, gamma glutamyl transferase, and bicarbonate increased with age. Mean corpuscular hemoglobin concentration, cellular hemoglobin concentration mean, platelets, lymphocytes, phosphorus, albumin/globulin ratio, alkaline phosphatase, anion gap, and calculated osmolality decreased with age. Males had higher hemoglobin, hematocrit, red blood cell count, mean corpuscular hemoglobin concentration, white blood cell count, heterophils, Foa-Kurloff cells, alanine aminotransferase, and bicarbonate and lower mean corpuscular volume, red blood cell distribution width, platelets, mean platelet volume, eosinophils, total protein, albumin, globulin, cholesterol, potassium, anion gap, calculated osmolality, and iron compared to females. Establishing age and sex differences in hematologic and biochemical parameters of Dunkin Hartley guinea pigs provides valuable insight into their physiology to better evaluate diagnostics and experimental results.

Abstract:

Abstract. Title of the paper: Biotechnical control of reproduction in sows and the opportunity of a hidden change of gene frequency for estrous behaviour Since more than three decades hormones have been used for the synchronisation of ovulation in sows often combined with timed artificial insemination. This will be done without considering the estrous behaviour. A selection with regard to symptoms of standing behaviour is not necessary. Regarding to that it is possible to breed pigs that are not suitable for that purpose. The aim of this study was to examine the opportunity of hidden gene frequency changes in the genetics of pigs in two herds of German Landrace. This will be shown by the example of the standing behaviour and especially the time they need to come in heat after weaning of the piglets. The basic datas was provided by two pig-breeding farms. The calculation is based only on the information to the 2nd and 3rd litter of these sows in order to reduce the influence of selection. It is assumed that the path of heredity is monofacturial. Only pigs with the genotype “aa” will be negatively affected. At the beginning the father of boars (SS) is the sole carrier of the defect gene. The gene frequency, frequency for hom*ozygotes and the correct number of concerned pigs after 10 generations will be calculated for a more accurate description of the expansion of this gene. The gene frequency results in the balance of 0.4 %. This figure has already been achieved after eight generations. According to this value the frequency of hom*ozygotes is 0.0017 % on all paths of heredity. After ten generations the population will contain 0.0093 animals of the hom*ozygot recessive genotype “aa”. There will be 0.0055 male and 0.0038 female pigs of the negative genotype. Finally, no negative effect of the usage of hormones for the synchronisation of ovulation in pigs has been proved.

Abstract:

Relaxin is a polypeptide hormone secreted by male and female reproductive tissues to facilitate spermatozoa progression in the female tract and parturition. Relaxin secretions are found in the vicinity of oocytes and embryos, and exert their effects through membrane receptors, which have not yet been described in porcine embryos. Here, we determined the presence of RXFP1 and RXFP2 receptors in porcine gametes and embryo, and evaluated the developmental effects of porcine relaxin (pRLX; Yan et al. 2006 Reproduction 131, 943-950). Cumulus-oocyte complexes (COC) were aspirated from sows ovaries collected at a local abattoir. hom*ogeneous COC were selected for IVM (44 h) and fertilization (6 to 8 h). Presumptive zygotes were cultured in NCSU-23 + 0.4% BSA for up to 7 days. All procedures were done at 39°C, under 5% CO2 in a humidified atmosphere. Matured oocytes, BTS-diluted spermatozoa, and embryos were collected for gene expression studies. For developmental studies, COC were matured (experiment 1), or embryos cultured from the zygote stage (experiment 2) in the presence ofpRLX (0, 20, or 40 ng mL-1). In experiment 3, zygotes derived from oocytes matured in the presence of pRLX (40 ng mL-1) were cultured with pRLX (20 or 40 ng mL-1). The pRLX effects were assessed on cleaved embryos and blastocysts recorded on Days 2 and 7 postinsemination, respectively. The total cell numbers of Day-7 blastocysts were also evaluated. All data were analyzed using ANOVA. Gametes and embryos expressed RXFP1 and RXFP2 at both the mRNA and protein level. The amounts of both gene transcripts were higher in mature oocytes (metaphase II) compared with spermatozoa (P < 0.05). The RXFP1/2 mRNA ratios were in favor of RXFP2 in mature oocytes (0.9×), zygotes (0.8 ×), and cleaved embryos (0.8×), and for RXFP1 in spermatozoa (1.1 ×) and blastocysts (1.1 ×). A similar pattern during embryo development was revealed at the protein level, showing a higher RXFP2 fluorescence signal in cleaved embryos and a lower signal in blastocysts compared with RXFP1 protein. In experiment 1, COC exposed to 40 ng mL-1 pRLX resulted in fewer cleaved embryos (36 ± 4%) compared with controls (42 ± 5%, P < 0.05). Of the 40 ng mL-1 pRLX-derived cleaved embryos, a greater proportion developed to the blastocyst stage (38 ± 6%; P < 0.05) compared with control and 20 ng mL-1 pRLX-derived cleaved embryos (26 ± 4% and 17 ± 8%, respectively). In experiment 2, however, 40 ng mL-1 pRLX induced higher cleavage but lower blastocyst rates (51 ± 5% and 20 ± 4%, respectively) compared with the control group (37 ± 4% and 32 ± 7%, respectively) (P < 0.05). In experiment 3, the exposure of both oocytes and derived embryos did not affect the developmental rates (P > 0.05). Nevertheless, pRLX significantly increased the mean cell number of blastocysts in all experiments (P < 0.05). We concluded that pig embryos express RXFP1 and RXFP2 receptors, which may facilitate a role for pRLX during oocyte maturation and embryo development in the pig. This work was supported by the USDA-ARS Biophotonics Initiative project# 58-6402-3-0120 and the Mississippi Agricultural and Forestry Experiment Station (MAFES).

Abstract:

Sexual reproduction requires that the gamete carrying the male-derived haploid chromatin join with the gamete carrying the female-derived haploid chromatin during fertilization to produce the diploid zygote. To accomplish this feat, the sperm must not only meet the egg, it must recognize the egg and be recognized in turn by the egg, and in the end must enter and be engulfed by the egg. In this selective overview of gamete interactions that lead to fertilization, encounters of three kinds, followed by the finale of gamete fusion, are considered from the sperm's viewpoint, with particular emphasis on the mammalian species with the mouse as the principal model. The first encounter is with the zona pellucida of the egg, to whose surface the sperm must bind. Mouse sperm appear to have four binding sites for zona ligands. Three interact with sugar moieties of the oligosaccharide chains of the mouse zona glycoprotein ZP3; the fourth binds a peptide backbone arginine. Capacitation is not required for this encounter, but is obligate for the second encounter--induction of the acrosome reaction in the bound sperm. The acrosome reaction is an exocytotic process that makes available the enzymatic machinery needed for sperm penetration the zona which is the end point of a sequence of reactions directed by intracellular signalling systems. In mouse sperm, these systems are presumed to be activated by ligands on ZP3 binding to ligand-specific sperm receptors with consequent aggregation of receptors. No receptor has been identified with certainty, nor have candidates for putative ZP3 ligands been identified. Completion of the acrosome reaction allows the sperm to penetrate the zona and, bind to the egg plasma membrane, thereby completing the third encounter. In the mouse, a 94-kDa protein appears essential for this binding. In the guinea-pig, a sperm plasma membrane protein (formerly PH-30, now fertilin), is a strong candidate for the mediator of the fusion process by which the egg engulfs the sperm. Decondensation of the sperm chromatin reverses the remarkable packing of DNA organized by sperm protamines. Mitochondrial DNA is also engulfed by the egg; the question of whether this DNA makes a small finite, or null, contribution to cytosolic inheritance is still in debate. The puzzles attending these encounters are presented as reminders of the intricacy and fascination, as well as of the vital necessity, of gamete interaction.

Abstract:

Osteopontin (OPN) is an acidic single-chain phosphorylated glycoprotein found in both the female and the male reproductive tract that is believed to facilitate reproduction. It was recently reported in swine that OPN improves in vitro fertilization (Hao et al. 2006 Biol. Reprod. 75, 726–733). In bovine, OPN improves the efficiency of in vitro embryo production, and it has been detected on the zona pellucida (ZP) of immature and mature oocytes (Monaco et al. 2007 J. Anim. Sci. 85, 529 abst). This study was designed to evaluate the gene expression of OPN (SPP1) in immature and mature swine cumulus cells and oocytes. Ovaries from a local slaughterhouse were collected on three different days. Half of the immature cumulus–oocyte complexes (COCs) from each day were vortexed in 1 mL TCM-199 HEPES containing 10 mg mL–1 hyaluronidase to separate immature cumulus cells, while the other half of the COCs were maturated for 40-42 h and then the cumulus cells were separated. RNA from cumulus cells and oocytes was extracted and genomic DNA was removed by DNase treatment. cDNA was synthesized using 100 ng of RNA and diluted 50% with DNase–RNase-free water. The RT-PCR product from cumulus cells and oocytes was run in a 2% agarose gel stained with ethidium bromide to verify the presence of SPP1. Relative mRNA abundance between immature and mature cumulus cells was assessed by SYBR green real-time RT–PCR run in triplicate using a 6-point twofold dilution standard curve. GAPDH was used as internal control. Primers for SPP1 and GAPDH were designed spanning an exon/exon junction. The presence of a single and specific PCR product was assessed by gel electrophoresis (a single band was expected at 100 and 90 bp, respectively, for SPP1 and GAPDH), dissociation curve, and sequencing. A t-test was used to assess differences between immature (n = 3) and mature (n = 3) cumulus cells. SPP1 was detected in agarose gel in both immature and mature oocytes and cumulus cells. Cumulus cells presented a Ct for SPP1 ranging from 20 (immature) to 29 (mature). Immature cumulus cells showed a 32-fold larger mRNA abundance compared to the mature cumulus cells (P < 0.05) when data were transformed using a standard curve. Results showed expression of SPP1 in porcine oocytes and cumulus cells. However, maturation significantly decreased the expression of SPP1 in cumulus cells. The presence of SPP1 mRNA in oocytes and cumulus cells and the larger mRNA abundance before maturation may suggest a role of this protein prior to maturation of oocytes. Additional studies will be required to determine the specific role of SPP1 in oocyte maturation in the pig.

Abstract:

Androgenic haploid embryos were originally produced for the study of certain aspects of early embryo development. The generation of androgenic haploid embryos allows us to better understand the complementary parental contribution to embryonic development, and to examine the effects of haploid development on gene expression. Because mare oocytes for research are scarce, the generation of heterospecific androgenic embryos could be useful to study aspects of the biology of early embryo development, or to identify genes and their variations or mutations that are responsible for reproduction-related problems in mares and stallions, which is of interest for the breeding industry. Therefore, the aim of this work was to study the capability of equine sperm to induce embryonic development after injection into an enucleated oocyte from a different species. Porcine cumulus-oocyte complexes (COC) were obtained from abattoir ovaries and placed in 100-µL drops in vitro maturation (IVM) medium for 42h. Cumulus cells were removed with hyaluronidase and vortexing. Then, mature oocytes were subjected to intracytoplasmic sperm injection (ICSI) with stallion frozen-thawed sem*n (according to Rodriguez et al. 2015). Immediately after the last injection, the zona pellucida of injected oocytes was removed with protease treatment, the oocytes were treated with cytochalasin B, and the metaphase II enucleated with a 20-µm micropipette. Finally, embryos were placed in culture medium (SOF) in plates with the well-of-the-well (WOW) system. As control treatment, non-enucleated pig oocytes were injected with stallion (CE) and boar (CC) sem*n. At Day 4, embryos were evaluated for cleavage and number of blastomeres, and stained with Hoechst 33342 to verify the presence of DNA in each blastomere under the UV light. Embryos were stored for future PCR studies to validate the presence of equine DNA. Data were analysed by chi-squared test to compare the cleavage of both controls with the androgenic embryos. From a total of 53 androgenic haploid embryos, the cleavage rate was 62% (33/53). Embryos were cleaved in 2 to 4 cells in 72.7%, 5 to 8 cells in 18.2%, and 9+ cells in 9.1% at Day 4. Presence of DNA in all blastomeres was observed in 60.6% (20/33) of the androgenic haploid embryos, while 21.2% (7/33) of the embryos had 10 to 50% of blastomeres with DNA, and 18.6% (6/33) of the embryos did not have DNA in their blastomeres. The ICSI control embryos cleaved in 45.3% (34/75) and 64.9% (98/151) for groups CC and CE, respectively. Cleavage rates in control CE were significantly higher than those in control CC (P&lt;0.004). No statistical difference was observed in the control groups versus androgenic embryos. This preliminary results showed that a heterospecific ooplasm can be successfully used to allow an equine sperm DNA to decondense and to develop, even in absence of the female counterpart. Using this method, copies of a single sperm DNA can be produced to potentially evaluate individual aspects of early embryo development concerning the male contribution. This is the first report of successful androgenic embryos using a heterospecific oocyte to create copies of a horse sperm DNA.

Abstract:

Abstract Background MicroRNAs (miRNAs) are small non-coding RNAs playing vital roles in regulating posttranscriptional gene expression. Elucidating the expression regulation of miRNAs underlying pig testis development will contribute to a better understanding of boar fertility and spermatogenesis. Results In this study, miRNA expression profile was investigated in testes of Duroc and Meishan boars at 20, 75, and 270 days of age by high-throughput sequencing. Forty-five differentially expressed miRNAs were identified from testes of Duroc and Meishan boars before and after puberty. Integrated analysis of miRNA and mRNA profiles predicted many miRNA-mRNA pairs. Gene ontology and biological pathway analyses revealed that predicted target genes of ssc-mir-423-5p, ssc-mir-34c, ssc-mir-107, ssc-mir-196b-5p, ssc-mir-92a, ssc-mir-320, ssc-mir-10a-5p, and ssc-mir-181b were involved in sexual reproduction, male gamete generation, and spermatogenesis, and GnRH, Wnt, and MAPK signaling pathway. Four significantly differentially expressed miRNAs and their predicted target genes were validated by quantitative real-time polymerase chain reaction, and phospholipase C beta 1 (PLCβ1) gene was verified to be a target of ssc-mir-423-5p. Conclusions This study provides an insight into the functional roles of miRNAs in testis development and spermatogenesis and offers useful resources for understanding differences in sexual function development caused by the change in miRNAs expression between Duroc and Meishan boars.

Abstract:

AbstractBoar taint is an unpleasant odor in male pig meat, mainly caused by androstenone, skatole, and indole, which are deposited in the fat tissue. Piglet castration is the most common practice to prevent boar taint. However, castration is likely to be banished in a few years due to animal welfare concerns. Alternatives to castration, such as genetic selection, have been assessed. Androstenone and skatole have moderate to high heritability, which makes it feasible to select against these compounds. This review presents the latest results obtained on genetic selection against boar taint, on correlation with other traits, on differences in breeds, and on candidate genes related to boar taint. QTLs for androstenone and skatole have been reported mainly on chromosomes 6, 7, and 14. These chromosomes were reported to contain genes responsible for synthesis and degradation of androstenone and skatole. A myriad of work has been done to find markers or genes that can be used to select animals with lower boar taint. The selection against boar taint could decrease performance of some reproduction traits. However, a favorable response on production traits has been observed by selecting against boar taint. Selection results have shown that it is possible to reduce boar taint in few generations. In addition, modifications in diet and environment conditions could be associated with genetic selection to reduce boar taint. Nevertheless, costs to measure and select against boar taint should be rewarded with incentives from the market; otherwise, it would be difficult to implement genetic selection.

Abstract:

As the name suggests, the humanities is largely a study of the human condition, in which history sits as a discipline concerned with the past. Environmental history is a new field that brings together scholars from a range of disciplines to consider the changing relationships between humans and the environment over time. Critiques of anthropocentrism that place humans at the centre of the universe or make assessments through an exclusive human perspective provide a challenge to scholars to rethink our traditional biases against the nonhuman world. The movement towards nonhumanism or posthumanism, however, does not seem to have had much of an impression on history as a discipline. What would a nonhumanist history look like if we re-centred the historical narrative around pigs? There are histories of pigs as food (see for example, The Cambridge History of Food which has a chapter on “Hogs”). There are food histories that feature pork in terms of its relationship to multiethnic identity (such as Donna Gabaccia’s We Are What We Eat) and examples made of pigs to promote ethical eating (Singer). Pigs are central to arguments about dietary rules and what motivates them (Soler; Dolander). Ancient pig DNA has also been employed in studies on human migration and colonisation (Larson et al.; Durham University). Pigs are also widely used in a range of products that would surprise many of us. In 2008, Christien Meindertsma spent three years researching the products made from a single pig. Among some of the more unexpected results were: ammunition, medicine, photographic paper, heart valves, brakes, chewing gum, porcelain, cosmetics, cigarettes, hair conditioner and even bio diesel. Likewise, Fergus Henderson, who coined the term ‘nose to tail eating’, uses a pig on the front cover of the book of that name to suggest the extraordinary and numerous potential of pigs’ bodies. However, my intention here is not to pursue a discussion of how parts of their bodies are used, rather to consider a reorientation of the historical narrative to place pigs at the centre of stories of our co-evolution, in order to see what their history might say about humans and our relationships with them. This is underpinned by recognition of the inter-relationality of humans and animals. The relationships between wild boar and pigs with humans has been long and diverse. In a book exploring 10,000 years of interaction, Anton Ervynck and Peter Rowley-Conwy argue that pigs have been central to complex cultural developments in human societies and they played an important role in human migration patterns. The book is firmly grounded within the disciplines of zoology, anthropology and archaeology and contributes to an understanding of the complex and changing relationship humans have historically shared with wild boar and domestic pigs. Naturalist Lyall Watson also explores human/pig relationships in The Whole Hog. The insights these approaches offer for the discipline of history are valuable (although overlooked) but, more importantly, such scholarship also challenges a humanist perspective that credits humans exclusively with historical change and suggests, moreover, that we did it alone. Pigs occupy a special place in this history because of their likeness to humans, revealed in their use in transplant technology, as well as because of the iconic and paradoxical status they occupy in our lives. As Ervynck and Rowley-Conwy explain, “On the one hand, they are praised for their fecundity, their intelligence, and their ability to eat almost anything, but on the other hand, they are unfairly derided for their apparent slovenliness, unclean ways, and gluttonous behaviour” (1). Scientist Niamh O’Connell was struck by the human parallels in the complex social structures which rule the lives of pigs and people when she began a research project on pig behaviour at the Agricultural Research Institute at Hillsborough in County Down (Cassidy). According to O’Connell, pigs adopt different philosophies and lifestyle strategies to get the most out of their life. “What is interesting from a human perspective is that low-ranking animals tend to adopt one of two strategies,” she says. “You have got the animals who accept their station in life and then you have got the other ones that are continually trying to climb, and as a consequence, their life is very stressed” (qtd. in Cassidy). The closeness of pigs to humans is the justification for their use in numerous experiments. In the so-called ‘pig test’, code named ‘Priscilla’, for instance, over 700 pigs dressed in military uniforms were used to study the effects of nuclear testing at the Nevada (USA) test site in the 1950s. In When Species Meet, Donna Haraway draws attention to the ambiguities and contradictions promoted by the divide between animals and humans, and between nature and culture. There is an ethical and critical dimension to this critique of human exceptionalism—the view that “humanity alone is not [connected to the] spatial and temporal web of interspecies dependencies” (11). There is also that danger that any examination of our interdependencies may just satisfy a humanist preoccupation with self-reflection and self-reproduction. Given that pigs cannot speak, will they just become the raw material to reproduce the world in human’s own image? As Haraway explains: “Productionism is about man the tool-maker and -user, whose highest technical production is himself […] Blinded by the sun, in thrall to the father, reproduced in the sacred image of the same, his rewards is that he is self-born, an auto telic copy. That is the mythos of enlightenment and transcendence” (67). Jared Diamond acknowledges the mutualistic relationship between pigs and humans in Guns, Germs and Steel and the complex co-evolutionary path between humans and domesticated animals but his account is human-centric. Human’s relationships with pigs helped to shape human history and power relations and they spread across the world with human expansion. But questioning their utility as food and their enslavement to this cause was not part of the account. Pigs have no voice in the histories we write of them and so they can appear as passive objects in their own pasts. Traces of their pasts are available in humanity’s use of them in, for example, the sties built for them and the cooking implements used to prepare meals from them. Relics include bones and viruses, DNA sequences and land use patterns. Historians are used to dealing with subjects that cannot speak back, but they have usually left ample evidence of what they have said. In the process of writing, historians attempt to perform the miracle, as Curthoys and Docker have suggested, of restoration; bringing the people and places that existed in the past back to life (7). Writing about pigs should also attempt to bring the animal to life, to understand not just their past but also our own culture. In putting forward the idea of an alternative history that starts with pigs, I am aware of both the limits to such a proposal, and that most people’s only contact with pigs is through the meat they buy at the supermarket. Calls for a ban on intensive pig farming (RSPCA, ABC, AACT) might indeed have shocked people who imagine their dinner comes from the type of family farm featured in the movie Babe. Baby pigs in factory farms would have been killed a long time before the film’s sheep dog show (usually at 3 to 4 months of age). In fact, because baby pigs do grow so fast, 48 different pigs were used to film the role of the central character in Babe. While Babe himself may not have been aware of the relationship pigs generally have to humans, the other animals were very cognisant of their function. People eat pigs, even if they change the name of the form it takes in order to do so:Cat: You know, I probably shouldn’t say this, but I’m not sure if you realize how much the other animals are laughing at you for this sheep dog business. Babe: Why would they do that? Cat: Well, they say that you’ve forgotten that you’re a pig. Isn't that silly? Babe: What do you mean? Cat: You know, why pigs are here. Babe: Why are any of us here? Cat: Well, the cow’s here to be milked, the dogs are here to help the Boss's husband with the sheep, and I’m here to be beautiful and affectionate to the boss. Babe: Yes? Cat: [sighs softly] The fact is that pigs don’t have a purpose, just like ducks don’t have a purpose. Babe: [confused] Uh, I—I don’t, uh ... Cat: Alright, for your own sake, I’ll be blunt. Why do the Bosses keep ducks? To eat them. So why do the Bosses keep a pig? The fact is that animals don’t seem to have a purpose really do have a purpose. The Bosses have to eat. It’s probably the most noble purpose of all, when you come to think about it. Babe: They eat pigs? Cat: Pork, they call it—or bacon. They only call them pigs when they’re alive (Noonan). Babe’s transformation into a working pig to round up the sheep makes him more useful. Ferdinand the duck tried to do the same thing by crowing but was replaced by an alarm clock. This is a common theme in children’s stories, recalling Charlotte’s campaign to praise Wilbur the pig in order to persuade the farmer to let him live in E. B. White’s much loved children’s novel, Charlotte’s Web. Wilbur is “some pig”, “terrific”, “radiant” and “humble”. In 1948, four years before Charlotte’s Web, White had published an essay “Death of a Pig”, in which he fails to save a sick pig that he had bought in order to fatten up and butcher. Babe tried to present an alternative reality from a pig’s perspective, but the little pig was only spared because he was more useful alive than dead. We could all ask the question why are any of us here, but humans do not have to contemplate being eaten to justify their existence. The reputation pigs have for being filthy animals encourages distaste. In another movie, Pulp Fiction, Vincent opts for flavour, but Jules’ denial of pig’s personalities condemns them to insignificance:Vincent: Want some bacon? Jules: No man, I don’t eat pork. Vincent: Are you Jewish? Jules: Nah, I ain’t Jewish, I just don’t dig on swine, that’s all. Vincent: Why not? Jules: Pigs are filthy animals. I don’t eat filthy animals. Vincent: Bacon tastes gooood. Pork chops taste gooood. Jules: Hey, sewer rat may taste like pumpkin pie, but I’d never know ’cause I wouldn’t eat the filthy motherf*cker. Pigs sleep and root in sh*t. That’s a filthy animal. I ain’t eat nothin’ that ain’t got sense enough to disregard its own feces [sic]. Vincent: How about a dog? Dogs eats its own feces. Jules: I don’t eat dog either. Vincent: Yeah, but do you consider a dog to be a filthy animal? Jules: I wouldn’t go so far as to call a dog filthy but they’re definitely dirty. But, a dog’s got personality. Personality goes a long way. Vincent: Ah, so by that rationale, if a pig had a better personality, he would cease to be a filthy animal. Is that true? Jules: Well we’d have to be talkin’ about one charming motherf*ckin’ pig. I mean he’d have to be ten times more charmin’ than that Arnold on Green Acres, you know what I’m sayin’? In the 1960s television show Green Acres, Arnold was an exceptional pig who was allowed to do whatever he wanted. He was talented enough to write his own name and play the piano and his attempts at painting earned him the nickname “Porky Picasso”. These talents reflected values that are appreciated, and so he was. The term “pig” is, however, chiefly used a term of abuse, however, embodying traits we abhor—gluttony, obstinence, squealing, foraging, rooting, wallowing. Making a pig of yourself is rarely honoured. Making a pig of the humanities, however, could be a different story. As a historian I love to forage, although I use white gloves rather than a snout. I have rubbed my face and body on tree trunks in the service of forestry history and when the temperature rises I also enjoy wallowing, rolling from side to side rather than drawing a conclusion. More than this, however, pigs provide a valid means of understanding key historical transitions that define modern society. Significant themes in modern history—production, religion, the body, science, power, the national state, colonialism, gender, consumption, migration, memory—can all be understood through a history of our relationships with pigs. Pigs play an important role in everyday life, but their relationship to the economic, social, political and cultural matters discussed in general history texts—industrialisation, the growth of nation states, colonialism, feminism and so on—are generally ignored. However “natural” this place of pigs may seem, culture and tradition profoundly shape their history and their own contribution to those forces has been largely absent in history. What, then, would the contours of such a history that considered the intermeshing of humans and pigs look like? The intermeshing of pigs in early human history Agricultural economies based on domestic animals began independently in different parts of the world, facilitating increases in population and migration. Evidence for long-term genetic continuity between modern and ancient Chinese domestic pigs has been established by DNA sequences. Larson et al. have made an argument for five additional independent domestications of indigenous wild boar populations: in India, South East Asia and Taiwan, which they use to develop a picture of both pig evolution and the development and spread of early farmers in the Far East. Domestication itself involves transformation into something useful to animals. In the process, humans became transformed. The importance of the Fertile Crescent in human history has been well established. The area is attributed as the site for a series of developments that have defined human history—urbanisation, writing, empires, and civilisation. Those developments have been supported by innovations in food production and animal husbandry. Pig, goats, sheep and cows were all domesticated very early in the Fertile Crescent and remain four of the world’s most important domesticated mammals (Diamond 141). Another study of ancient pig DNA has concluded that the earliest domesticated pigs in Europe, believed to be descended from European wild boar, were introduced from the Middle East. The research, by archaeologists at Durham University, sheds new light on the colonisation of Europe by early farmers, who brought their animals with them. Keith Dobney explains:Many archaeologists believe that farming spread through the diffusion of ideas and cultural exchange, not with the direct migration of people. However, the discovery and analysis of ancient Middle Eastern pig remains across Europe reveals that although cultural exchange did happen, Europe was definitely colonised by Middle Eastern farmers. A combination of rising population and possible climate change in the ‘fertile crescent’, which put pressure on land and resources, made them look for new places to settle, plant their crops and breed their animals and so they rapidly spread west into Europe (ctd in ScienceDaily). Middle Eastern farmers colonised Europe with pigs and in the process transformed human history. Identity as a porcine theme Religious restrictions on the consumption of pigs come from the same area. Such restrictions exist in Jewish dietary laws (Kashrut) and in Muslim dietary laws (Halal). The basis of dietary laws has been the subject of much scholarship (Soler). Economic and health and hygiene factors have been used to explain the development of dietary laws historically. The significance of dietary laws, however, and the importance attached to them can be related to other purposes in defining and expressing religious and cultural identity. Dietary laws and their observance may have been an important factor in sustaining Jewish identity despite the dispersal of Jews in foreign lands since biblical times. In those situations, where a person eats in the home of someone who does not keep kosher, the lack of knowledge about your host’s ingredients and the food preparation techniques make it very difficult to keep kosher. Dietary laws require a certain amount of discipline and self-control, and the ability to make distinctions between right and wrong, good and evil, pure and defiled, the sacred and the profane, in everyday life, thus elevating eating into a religious act. Alternatively, people who eat anything are often subject to moral judgments that may also lead to social stigmatisation and discrimination. One of the most powerful and persuasive discourses influencing current thinking about health and bodies is the construction of an ‘obesity epidemic’, critiqued by a range of authors (see for example, Wright & Harwood). As omnivores who appear indiscriminate when it comes to food, pigs provide an image of uncontrolled eating, made visible by the body as a “virtual confessor”, to use Elizabeth Grosz’s term. In Fat Pig, a production by the Sydney Theatre Company in 2006, women are reduced to being either fat pigs or shrieking shallow women. Fatuosity, a blog by PhD student Jackie Wykes drawing on her research on fat and sexual subjectivity, provides a review of the play to describe the misogyny involved: “It leaves no options for women—you can either be a lovely person but a fat pig who will end up alone; or you can be a shrill bitch but beautiful, and end up with an equally obnoxious and shallow male counterpart”. The elision of the divide between women and pigs enacted by such imagery also creates openings for new modes of analysis and new practices of intervention that further challenge humanist histories. Such interventions need to make visible other power relations embedded in assumptions about identity politics. Following the lead of feminists and postcolonial theorists who have challenged the binary oppositions central to western ideology and hierarchical power relations, critical animal theorists have also called into question the essentialist and dualist assumptions underpinning our views of animals (Best). A pig history of the humanities might restore the central role that pigs have played in human history and evolution, beyond their exploitation as food. Humans have constructed their story of the nature of pigs to suit themselves in terms that are specieist, racist, patriarchal and colonialist, and failed to grasp the connections between the oppression of humans and other animals. The past and the ways it is constructed through history reflect and shape contemporary conditions. In this sense, the past has a powerful impact on the present, and the way this is re-told, therefore, also needs to be situated, historicised and problematicised. The examination of history and society from the standpoint of (nonhuman) animals offers new insights on our relationships in the past, but it might also provide an alternative history that restores their agency and contributes to a different kind of future. As the editor of Critical Animals Studies, Steve Best describes it: “This approach, as I define it, considers the interaction between human and nonhuman animals—past, present, and future—and the need for profound changes in the way humans define themselves and relate to other sentient species and to the natural world as a whole.” References ABC. “Changes to Pig Farming Proposed.” ABC News Online 22 May 2010. 10 Aug. 2010 http://www.abc.net.au/news/stories/2010/05/22/2906519.htm Against Animal Cruelty Tasmania. “Australia’s Intensive Pig Industry: The Intensive Pig Industry in Australia Has Much to Hide.” 10 Sep. 2010 http://www.aact.org.au/pig_industry.htm Babe. Dir. Chris Noonan. Universal Pictures, 1995. Best, Steven. “The Rise of Critical Animal Studies: Putting Theory into Action and Animal Liberation into Higher Education.” Journal for Critical Animal Studies 7.1 (2009): 9-53. Cassidy, Martin. “How Close are Pushy Pigs to Humans?”. BBC News Online 2005. 10 Sep. 2010 http://news.bbc.co.uk/2/hi/uk_news/northern_ireland/4482674.stmCurthoys, A., and Docker, J. “Time Eternity, Truth, and Death: History as Allegory.” Humanities Research 1 (1999) 10 Sep. 2010 http://www.anu.edu.au/hrc/publications/hr/hr_1_1999.phpDiamond, Jared. Guns, Germs and Steel: The Fates of Human Societies. New York: W. W. Norton, 1999. Dolader, Miguel-Àngel Motis. “Mediterranean Jewish Diet and Traditions in the Middle Ages”. Food: A Culinary History. Eds. Jean-Louis Flandrin and Massimo Montanari. Trans. Clarissa Botsford, Arthus Golhammer, Charles Lambert, Frances M. López-Morillas and Sylvia Stevens. New York: Columbia UP, 1999. 224-44. Durham University. “Chinese Pigs ‘Direct Descendants’ of First Domesticated Breeds.” ScienceDaily 20 Apr. 2010. 29 Aug. 2010 http://www.sciencedaily.com/releases/2010/04/100419150947.htm Gabaccia, Donna R. We Are What We Eat: Ethnic Food and the Making of Americans. Cambridge: Harvard University Press, 1998. Grosz, Elizabeth. Volatile Bodies: Toward a Corporeal Feminism. St Leonards: Allen & Unwin, 1994. Haraway, D. “The Promises of Monsters: A Regenerative Politics for Inappropriate/d Others.” The Haraway Reader. New York: Routledge, 2005. 63-124. Haraway, D. When Species Meet: Posthumanities. 3rd ed. London: University of Minnesota Press, 2008. Henderson, Fergus. Nose to Tail Eating: A Kind of British Cooking. London: Bloomsbury, 2004. Kiple, Kenneth F., Kriemhild Coneè Ornelas. Cambridge History of Food. Cambridge: Cambridge University Press, 2000. Larson, G., Ranran Liu, Xingbo Zhao, Jing Yuan, Dorian Fuller, Loukas Barton, Keith Dobney, Qipeng Fan, Zhiliang Gu, Xiao-Hui Liu, Yunbing Luo, Peng Lv, Leif Andersson, and Ning Li. “Patterns of East Asian Pig Domestication, Migration, and Turnover Revealed by Modern and Ancient DNA.” Proceedings of the National Academy of Sciences, United States 19 Apr. 2010. 10 Sep. 2010 http://www.pnas.org/cgi/content/full/0912264107/DCSupplemental Meindertsma, Christien. “PIG 05049. Kunsthal in Rotterdam.” 2008. 10 Sep. 2010 http://www.christienmeindertsma.com/index.php?/books/pig-05049Naess, A. “The Shallow and the Deep, Long-Range Ecology Movement.” Inquiry 16 (1973): 95-100. Needman, T. Fat Pig. Sydney Theatre Company. Oct. 2006. Noonan, Chris [director]. “Babe (1995) Memorable Quotes”. 10 Sep. 2010 http://www.imdb.com/title/tt0112431/quotes Plumwood, V. Feminism and the Mastery of Nature. London: Routledge, 1993. Pulp Fiction. Dir. Quentin Tarantino. Miramax, 1994. RSPCA Tasmania. “RSPCA Calls for Ban on Intensive Pig Farming.” 10 Sep. 2010 http://www.rspcatas.org.au/press-centre/rspca-calls-for-a-ban-on-intensive-pig-farming ScienceDaily. “Ancient Pig DNA Study Sheds New Light on Colonization of Europe by Early Farmers” 4 Sep. 2007. 10 Sep. 2010 http://www.sciencedaily.com/releases/2007/09/070903204822.htm Singer, Peter. “Down on the Family Farm ... or What Happened to Your Dinner When it was Still an Animal.” Animal Liberation 2nd ed. London: Jonathan Cape, 1990. 95-158. Soler, Jean. “Biblical Reasons: The Dietary Rules of the Ancient Hebrews.” Food: A Culinary History. Eds. Jean-Louis Flandrin and Massimo Montanari. Trans. Clarissa Botsford, Arthus Golhammer, Charles Lambert, Frances M. López-Morillas and Sylvia Stevens. New York: Columbia University Press, 1999. 46-54. Watson, Lyall. The Whole Hog: Exploring the Extraordinary Potential of Pigs. London: Profile, 2004. White, E. B. Essays of E. B. White. London: HarperCollins, 1979. White, E. B. Charlotte’s Web. London: HarperCollins, 2004. Wright, J., and V. Harwood. Eds. Biopolitics and the ‘Obesity Epidemic’. New York: Routledge, 2009. Wykes, J. Fatuosity 2010. 29 Aug. 2010 http://www.fatuosity.net

Abstract:

IntroductionAccording to Friedrich Nietzsche, the death of Man follows the death of God. Man as a concept must be overcome. Yet Nietzsche extends humanism’s jargon of creativity that privileges Man over animal. To truly overcome the notion of Man, one must undercome Man, in other words go below Man. Once undercome, creativity devolves into a type of building and unbuilding, affording art the ability to conceive of the subject emptied of divine creation. This article will examine how Man is unbuilt in three works by three different artists: Francis Bacon’s “Study of a Baboon” (1953), Jan Švankmajer’s Darkness, Light, Darkness (1989), and Patricia Piccinini’s “The Young Family” (2002). All three artists evoke the animalistic in their depiction of what could be called the sub-subject, a diminished agent. Unbuilding the subject becomes the basis for building the sub-subject in these depictions of the human remainder. Man, from this vantage, will be examined as a cultural construct. Man largely means human, yet the Renaissance concept favoured a certain type of powerful male. Instead of rescuing Man, Bacon, Švankmajer and Piccinini, present the remnants of the human amidst the animal rather than the human subject detached from the animal. Such works challenge humanism, expressed in Giorgio Vasari’s analysis of art and creativity as indicative of Man’s closeness to the divine, which in a strange way, is extended in Nietzsche’s writings. These artists dismantle and build a subhuman form of subjectivity and thereby provide a challenge to traditional conceptions of creativity that historically favour Man as the creator beneath only God Himself. In the course of this article, I explore the violence of Bacon’s painted devolution, the deflationary animation of Švankmajer and Piccinini’s subhuman tenderness. I do not argue that we must abandon humanism altogether as there are a multiplicity of humanisms, or attempt to invalidate all the various posthumanisms, transhumanisms and antihumanisms. Rather, I attempt to show that Nietzsche’s posthumanism is a suprahumanism and that one possible way to frame the death of Man is through undercoming Man. Art, held in high esteem by Renaissance humanism, becomes a vehicle to imagine and engage with subhuman subjectivity.What Is Humanism? Humanism has numerous connotations from designating atheism to celebrating culture to privileging humans above other animals. The type of humanism I am interested in is not secular humanism, but rather humanism that celebrates and conceptualises Man’s place in the universe and does so through accentuating his (and I mean his given humanism’s often sexist, masculinist history) creativity and intellectual power. This celebration of creativity depends in part on a type of religious view, where Man is at the centre of God’s design. Such a view holds that Man’s power to shape nature’s materials resembles God. This type of humanism remains today but usually in a more humbled form, enfeebled by the scientific realisations that characterised the Enlightenment, namely the realisation that Man was not the centre of God’s universe. The Enlightenment is sometimes characterised as the birth of modern humanism, where the human subject undergoes estrangement from his surroundings through the conceptualisation of the subject–object division, and gains control over nature. A common narrative is that the subject’s autonomy and power came to extend to art itself, which in turn, became valued as possessing its own aesthetic legitimacy and yet also becoming an alienated commodity. Yet Cary Wolfe, in What Is Posthumanism?, echoes Michel Foucault’s claim that the Enlightenment could be viewed in tension to humanism (“Introduction” n.p.). Indeed, the Enlightenment’s creation of modern science would come to seriously challenge any view of humanity’s privileged status in this world. In contrast, Renaissance humanism conceived of Man as the centrepiece of God’s design and gifted with artistic creation and the ability to uncover truth. Renaissance HumanismRenaissance humanism is encapsulated by Vasari’s preface to The Lives of the Artists. In his preface, Vasari contends that God was the first artist, being both a painter and sculptor: God on High, having created the great body of the world and having decorated the heavens with its brightest lights, descended with His intellect further down into the clarity of the atmosphere and the solidity of the earth, and, shaping man, discovered in the pleasing invention of things the first form of sculpture and painting. (3)Interestingly, God discovers creation, which is a type of decoration, where the skies are decorated with bright lights—the stars. Giving colour, light and shade to the world and heavens, qualifies God as a painter. The human body, according to Vasari, is sculpted by God, which in turn inspires artists to depict the human form. Art and design—God’s design—is thereby ‘at the origin of all things’ and not merely painting and sculpture, though the reality we know is still the product of God’s painting and sculpture. According to Vasari, God privileges Man not for his intellect per se, but by bestowing him with the ability of creation and design. Indeed, creativity and design are for Vasari a part of all intellectual discovery. Intellect is the mode of discovering design, which for Vasari, is also creation. Vasari claims “that divine light infused in us by a special act of grace which has not only made us superior to other animals but even similar, if it is permitted to say so, to God Himself” (4). God is more than just a maker, he is a creator with an aesthetic sense. All intellectual human endeavours, claims Vasari, are aesthetic and creative, in their comprehension of God’s design of the world. Vasari’s emphasis on design became outmoded as Renaissance humanism was challenged by the Enlightenment’s interest in humans and other animals as machines. However, evolution challenges even some mechanistic understandings of the human subject, which sometimes presupposed that the human-machine had a maker, as with William Paley’s watchmaker theory. As Richard Dawkins put it in The Blind Watchmaker, nature “has no vision, no foresight, no sight at all. If [evolution] can be said to play the role of the watchmaker in nature, it is the blind watchmaker” (“Chapter One: Explaining the Very Improbable” n.p.). No longer was God’s universe designed for Man’s comprehension and appreciation, foretelling humanity’s own potential extinction.Man and God’s DeathThe idea that humanity was created by blind processes raises the question of what sort of depiction of the human subject is possible after the death of God and the Enlightenment’s tendency toward disenchantment? An art and self-understanding founded on atheism would be in sharp distinction to Vasari’s characterisation of the nature as an artwork coloured by the divine painter and sculptor in the heavens. Man’s creativity and design are, for the Renaissance humanist, part of discovery, the embodied realisations and iterations of the Platonic realm of divine forms. But such designs, wondrous for Vasari, can be viewed as shadows without origin in a post-God world. In Vasari, Platonism is still present where the artist’s creation becomes a way of discerning the origin of all forms, God himself. Yet, without divine origin, these forms are no longer discoveries and the possibility emerges that they are not even creations, emptied of the divine meaning that gave Man’s creative and scientific work value. Nietzsche understood that the loss of God called for the revaluation of all values. This is why Nietzsche claims that God’s death signifies the death of Man. For Nietzsche, the last Man was such an iteration, a shadow of what man had been (Thus Spoke Zarathustra 9-10). The Post-Man, the Übermensch, is one who extends the human power of creation and evaluation. In Vasari, Man is a model created by God. Nietzsche extends this logic: Man is his own creation as is God Man’s model. Man is capable of self-construction and overcoming without the hindrance of the divine. This freedom unlocked by auto-creation renders Man capable of making himself God. As such, art remains a source of sacred power for Nietzsche since it is a process of creative evaluation. The sacred is affirmed against secular profanity. For Nietzsche, God must be envisaged as Dionysus, a God that Nietzsche claims takes on a human form in Greek festivals dedicated to creation and fecundity. Mankind, in order to continue to have value after God’s death, “must become gods”, must take the place of God (The Gay Science 120). Nietzsche, All-Too HumanistNietzsche begins a project of rethinking Man as a category. Yet there is much in common with Renaissance humanism generated by Nietzsche’s Dionysian belief in a merger between God and Man. Man is overcome by a stronger and more creative figure, that of the Übermensch. By comparing Nietzsche with Vasari we can understand just how humanist Nietzsche remained. Indeed, Nietzsche fervently admired the Renaissance as a rebirth of paganism. Such an assessment of the rebirth of pagan art and values can almost be found in Vasari himself. Vasari claimed that pagan art, far from being blasphemous, brought Man closer to the divine in a tribute to the creativity of God. Vasari’s criticism of Christianity is careful but present. Indeed, Vasari—in a way that anticipates Nietzsche’s view that secular sacrilege was merely an extension of Christian sacrilege—attacks Christian iconoclasm, noting that barbarians and Christians worked together to destroy sacred forms of art: not only did [early Christianity] ruin or cast to the ground all the marvellous statues, sculptures, paintings, mosaics, and ornaments of the false pagan gods, but it also did away with the memorials and testimonials to an infinite number of illustrious people, in whose honour statues and other memorials had been constructed in public places by the genius of antiquity. (5) In this respect, Vasari embodies the values Nietzsche so praised in the Italian Renaissance. Vasari emphasises the artistic creations that enshrine distinctions of value and social hierarchy. While Vasari continues Platonic notions that ideals exist before human creation, he nevertheless holds human creation as a realisation and embodiment of the ideal, which is not dissimilar to Nietzsche’s notion of divine embodiment. For Nietzsche and Vasari, Man is exulted when he can rise, like a god, above other men. Another possibility would be to lower Man to just another animal. One way to envision such a lowering would be to subvert the mode by which Man is deemed God-like. Art that engages with the death of Man helps conceptualise subhumanism and the way that the subject ceases to be raised above the animal. What follows are studies of artworks that unbuild the subject. Francis Bacon’s “Study of a Baboon”Francis Bacon’s work challenges the human subject by depicting nonhuman subjects, where the flesh is torn open and Man’s animal flesh is exposed. Sometimes Bacon does not merely disfigure the human form but violently abandons it to focus on animals that reveal animal qualities latent in the subject. Bacon’s “Study of a Baboon”, expresses a sense of human devolution: Man devolved to monkey. In the work, we see a baboon within an enclosure, sitting above a tree that simultaneously resembles a gothic shadow, a cross, and even a smear. The dark, cross-like tree may suggest the conquering of God by a baboon, a type of monkey, recalling the old slander of Darwin’s theory, namely that Darwinism entailed that humanity descended from monkeys (which Darwin’s theory does not claim). But far from victorious, the monkey is in a state of suffering. While the baboon is not crucified on or by the tree, suffering pervades the frame. Its head resembles some sort of skull. The body is faintly painted in a melancholy blue with smudges of purple and is translucent and ghostly—at once a lump of matter and a spectral absence. We do not see the baboon through the cage. Instead we see through the baboon at the cage. Indeed, its very physiology involves the encountering of trauma as the head of the baboon does not simply connect to the body but stabs through the body as a sharp bone, perhaps opaquely evoking the violence of evolution. Similarly, the baboon’s tail seems to stab through the tree. Its eye is an enlarged void and a pupil is indicated by a bluish white triangle splitting through the void. The tree has something of the menacing and looming quality of a shadow and there is a sense of wilderness confronted by death and entrapment, evoked through the background. The yellowy ground is suggestive of dead grass. While potentially gesturing to the psychical confusion and intensity of Vincent Van Gogh or Edvard Munch, the yellowed grass more likely evokes the empty, barren and hostile planes of the desert and contrasts with the darkened colours. The baboon sitting on the cross/tree may seem to have reached some sort of pinnacle but such a status is mocked by the tree that manages to continue outside the fence: the branches nightmarishly protrude through the fence to conquer the frame, which in turn furthers the sense of inescapable entrapment and threat. The baboon is thereby precluded from reaching a higher point on the tree, unable to climb the branches, and underscores the baboon’s confines. The painting is labelled a study, which may suggest it is unfinished. However, Bacon’s completed works preserve an unfinished quality. This unfinished quality conveys a sense in which Man and evolution are unfinished and that being finished in the sense of being completed is no longer possible. The idea that there can finished work of art, a work of art that preserves an eternal meaning, has been repeatedly subject to serious doubt, including by artists themselves. Indeed, Bacon’s work erases the potential for perfection and completion, and breaks down, through devolution, what has been achieved by Man and the forces that shaped him. The subject is lowered from that of human to that of a baboon and is therefore, by Vasari’s Renaissance reasoning, not a subject at all. Bacon’s sketch and study exist to evoke a sense of incompletion, involving pain without resolution. The animal state of pain is therefore married with existential entrapment and isolation as art ceases to express the Platonic ideal and aims to show the truth of the shadow—namely that humanity is without a God, a God that previously shed light on humanity’s condition and anchored the human subject. If there is a trace or echo of human nobility left, such a trace functions through the wild and violent quality of animal indignation. A scream of painful indignity is the last act approaching (or descending from) any dignity that is afforded. Jan Švankmajer’s Darkness, Light, DarknessAn even more extreme case of the subject no longer being the subject, of being broken and muted—so much so that animal protest is annulled—can be witnessed in Jan Švankmajer’s animated short Darkness, Light, Darkness. In the animation, green clay hands mould and form a human body in order to be part of it. But when complete, the human body is trapped, grotesquely out of proportion with its environment. The film begins in a darkened house. There is a knocking of the door, and then the first green hand opens the door and turns on the light. The hand falls to the floor, blindly making its way to another door on the opposite side of the house. The hand opens the door only for eyeballs to roll out. The eyes look around. The hand pushes its clay fingers against the eyeballs, and the eyeballs become attached to the fingers. Suddenly with sight, the hand is able to lift itself up. The hand discovers that another hand is knocking at the door. The first hand helps the second hand, and then goes to the window where a pair of ears are stuck together flapping like a moth. The hands work together and break the ears apart. The first hand, the one with eyes, attaches the ears to the second hand. Then a head with a snout, but missing eyes and ears, enters through the door. The hands pull the snout until it becomes a nose, suppressing and remoulding the animal until it becomes human. As with Bacon, the violence of evolution, of auto-construction is conveyed indirectly: in Bacon’s case, through painted devolution and, in the case of the claymation, through a violent construction based on mutilation and smashing body parts together.Although I have described only three minutes of the seven-minute film, it already presents an image of human construction devoid of art or divine design. Man, or rather the hands, become the blind watchman of evolution. The hands work contingently, with what they are provided. They shape themselves based on need. The body, after all, exists as parts, and the human body is made up of other life forms, both sustaining and being sustained by them. The hands work together, and sacrifice sight and hearing for the head. They tear off the ears and remove the eyes and give them to the head. Transcendence is exchanged for subsistence. The absurdity of this contingency becomes most apparent when the hands attempt to merge with the head, to be the head’s feet. Then the feet actually arrive and are attached to the head’s neck. The human subject in such a state is thereby deformed and incomplete. It is a frightened form, cowering when it hears banging at the door. It turns out that the banging is being produced by an angry erect penis pounding at the door. However, even this symbol of masculine potency is subdued, rendered harmless by the hands that splash a bucket of cold water on it. The introduction of the penis signifies the masculinist notions implicit in the term Man, but we only ever see the penis when it is flaccid. The human subject is able to be concluded when clay pours from both doors and the window. The hands sculpt the clay and make the body, which, when complete is oversized and barely fits within the house. The male subject is then trapped, cramped in a foetal position. With its head against the ceiling next to the light, breathing heavily, all it can do is turn out the light. The head opens its mouth either in horror or a state of exertion and gasps. The eyes bulge before one of the body’s hands turns switch, perhaps suggesting terror before death or simply the effort involved in turning off the light. Once completed and built, the human subject remains in the dark. Despite the evident quirky, playful humour, Švankmajer’s film reflects an exhaustion with art itself. Human life becomes clay comically finding its own form. For Vasari, the ideal of the human form is realised first by God and then by Man through marble; for Švankmajer it is green clay. He demotes man back to the substance for a God to mould but, as there is no God to breathe life into it and give form, there is just the body to imperfectly mould itself. The film challenges both Vasari’s humanism and the suprahumanism of Nietzschean spectacle. Instead of the self-generating power and radical interdependence and agency of Übermensch, Švankmajer’s sub-subject is Man undercome—man beneath as opposed to over man, man mocked by its ambition, and with no space to stand high. Švankmajer thereby realises the anti-Nietzschean potential inherent within cinema’s anti-spectacular nature. Antonin Artaud, who extends the aesthetics advanced by Nietzsche in The Birth of Tragedy, contrasts the theatre’s sense of animal life with cinema. Artaud observes that movies “murder us with second-hand reproductions […] filtered through machines” (84). Thus, films murder creative and animal power as film flattens life to a dead realm of reproduction. Continuing Jacques Derrida’s hauntological framing of the screen, the animation theorist Alan Cholodenko has argued that the screen implies death. Motion is dead and replaced by illusion, a recording relayed back to us. What renders cinema haunting also renders it hauntological. For Cholodenko, cinema’s animation challenges ontology and metaphysics by eschewing stable ontologies through a process that entails both presence and absence. As Cholodenko points out, all film is a type of animation and reanimation, of making images move that are not in fact moving. Thus, one can argue that the animated-animation (such as Švankmajer’s claymation) becomes a refinement of death, a Frankesteinian reanimation of dead material. Indeed, Darkness, Light, Darkness accentuates the presence of death with the green clay almost resembling putrefaction. The fingerprints on the clay accentuate a lack of life, for the autonomous and dead matter that constructs and shapes a dead body from seemingly severed body parts. Even the title of the film, Darkness, Light, Darkness reflects an experience of cinema as deflation rather than joyous spectacle. One goes to a darkened space, watches light flicker on a screen and then the light goes out again. The cartoonish motions of the hands and body parts in the film look only half alive and therefore seem half-dead. Made in the decaying Communist state of Czechoslovakia, Švankmajer’s film aptly acknowledges the deflation of cinema, reflecting that illumination—the light of God, is put out, or more specifically, switched off. With the light of God switched off, creation becomes construction and construction becomes reconstruction, filtered through cinema’s machine processes as framed through Cholodenko. Still, Švankmajer’s animation is not unsympathetic to the plight of the hands. We do see the body parts work together. When a vulgar, meaty, non-claymation tongue comes out through the door, it goes straight to the other door to let the teeth in. The teeth and tongue are aided by the hands to complete the face. Indeed, what they produce is a human being, which has some sense of coherence and success—a success enmeshed with failure and entrapment. Piccinini’s “The Young Family”Patricia Piccinini’s sculptural works offer a more tender approach to the subject, especially when her works focus on the nonhuman animal with human characteristics. Piccinini is interested in the combinations of the animal and the machine, so her ideas can be seen almost as transhuman, where the human is extended beyond humanism. Her work is based on connection and connectedness, but does not emphasise the humanist values of innovation and self-creation often inherent to transhumanism. Indeed, the emphasis on connection is distinct from the entrapment of Bacon’s baboon and Švankmajer’s clay human, which half lament freedom’s negation.The way that Piccinini preserves aspects of humanism within a framework of subhumanism is evident in her work “The Young Family”. The hypperrealistic sculpture depicts a humanoid pig form, flopped, presumably exhausted, as piglet-babies suckle on her nipples. The work was inspired by a scientific proposal for pigs to be genetically modified to provide organs for humans (“Educational Resource” 5). Such a transhuman setting frames a subhuman aesthetic. Care is taken to render the scene with sentiment but without a sense of the ideal, without perfection. One baby-piglet tenderly grasps its foot with both hands and stares with love at its mother. We see two piglets enthusiastically sucking their mother’s teat, while a third baby/piglet’s bottom is visible, indicating that there is a third piglet scrambling for milk. The mother gazes at us, with her naked mammalian body visible. We see her wrinkles and veins. There is some fur on her head and some hair on her eyebrows humanising her. Indeed, her eyes are distinctly human and convey affection. Affection seems to be a motif that carries through to the materials (carefully crafted by Piccinini’s studio). The affection displayed in the artwork is trans-special, emphasising that human tenderness is in fact mammalian tenderness. Such tenderness conflates the human, the nonhuman animal and the material out of which the humanoid creature and its young are constructed. The sub-agency brings together the young and the old by displaying the closeness of the family. Something of this sub-subjectivity is theorised in Malcolm Bull’s Anti-Nietzsche, where he contrasts Nietzsche’s idea of the Übermensch with the idea of the subhuman. Bull writes that subhumanism involves giving up on “becoming more than a man and think[ing] only of becoming something less” (n.p.; Chapter 2, sec. “The Subhuman”). Piccinini depicts vulnerability and tenderness with life forms that are properly speaking subhuman, and reject the displays of strength of Nietzsche’s suprahumanism or Vasari’s emphasis on art commemorating great men. But Piccinini’s subhumanism preserves enough humanism to understand art’s ability to encourage an ethics of nurturing. In this respect, her works offer an alternative to Bull’s subhumanism that aims, so Bull argues, to devalue art altogether. Instead, Piccinini affirms imagination, but through its ability to conjure new ways to perceive animal affection. The sub-subject thereby functions to reveal states of emotion common to mammals (including humans) and other animals. ConclusionThese three artists therefore convey distinct, if related and intersecting, ways of visualising the sub-subject: Bacon through animal suffering, Švankmajer through adaptation that ultimately leads to the agent’s entrapment, and Piccinini who, instead of marrying anti-humanism with the subhumanism (the procedure of Švankmajer, and Bacon), integrates aspects of transhumanism and Renaissance humanism into her subhuman vision. As such, these works present a realisation of how we might think of the going under of the human subject after Darwin, Nietzsche and the deaths of God, Man and the diminishment of creativity. Such works remain not only antithetical to Vasari’s humanism but also to Nietzsche’s suprahumanism. These artists use art’s power to humble—not through overpowering awe but through the visible breakdown of the human agent, speaking for and to the sub-subject. Such art, by unbuilding and dismantling the subject, draws on prehuman trajectories of evolution, and in the case of Piccinini, transhuman trajectories. Art ceases to be about the grandiose evocations of power. Rather, more modestly, these works build a connection between the human with other mammals. Acknowledgements I wish to acknowledge Daniel Canaris for his valuable insights into Christianity and the Italian Renaissance, Alan Cholodenko for providing copies of his works that were central to my interpretation of Švankmajer, and Rachel Franks and Simon Dwyer for their invaluable assistance and finding very helpful reviewers. References Artaud, Antonin. The Theatre and Its Double. New York: Grove P, 1958.Art Gallery of South Australia. “Educational Resource Patricia Piccinini.” Adelaide: Art Gallery of South Australia. 11 Dec. 2016 <https://www.artgallery.sa.gov.au/agsa/home/Learning/docs/Online_Resources/Piccinini_online_resource.pdf>.Bacon, Francis. “Head I.” 1948. Oil on Canvas. 100.3 x 74.9cm. ———. “Study of a Baboon.” 1953. Oil on Canvas. 198.3 x 137.3cm. Bull, Malcolm. Anti-Nietzsche. New York: Verso, 2011. Cholodenko, Alan. “First Principles of Animation.” Animating Film Theory. Ed. Karen Beckman. Duke UP, 2014. 98-110.———. “The Crypt, the Haunted House, of Cinema.” Cultural Studies Review 10.2 (2004): 99-113. Darkness, Light, Darkness. Jan Švankmajer, 1990. 35mm. Nietzsche, Friedrich. The Birth of Tragedy and Other Writings. Cambridge: Cambridge UP, 2007. ———. The Gay Science. Cambridge: Cambridge UP, 2007. ———. Thus Spoke Zarathustra. Cambridge: Cambridge UP, 2006.Piccinini, Patricia. “The Young Family.” 2002. Silicone, Polyurethane, Leather, Plywood, Human Hair, 80 x 150 x 110cm. Vasari, Giorgio. The Lives of Artists. Oxford: Oxford UP, 1998.Wolfe, Cary. What Is Posthumanism? Minneapolis: U of Minnesota P, 2010.