A de novo reciprocal chromosomal translocation t(3;6)(p14;q26) in the black Lucano pig

Reprod Dom Anim. 2020;55:677–682. wileyonlinelibrary.com/journal/rda © 2020 Blackwell Verlag GmbH 

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1 | INTRODUCTION

The most frequent chromosomal abnormality in pigs is the balanced reciprocal translocation (rcp; Ducos et al., 2008), which represents about 90% of the rearrangements described in literature (Rothschild & Ruvinsky, 2011). Up to date, over 190 different types of rcp have been identified, in particular in commercial breeds (Donaldson, Villagomez, Revay, Rezaei, & King, 2019; Raudsepp & Chowdhary, 2011).

The structural balanced autosomal aberrations escape nor-mal breeding selection because the carriers have nornor-mal body

conformation. These chromosomal abnormalities are often the cause of low fertility or sterility (Genualdo et al., 2018; Kociucka, Szczerbal, Bugaj, Orsztynowicz, & Switonski, 2014; Szczerbal & Switonski, 2016; Villagomez & Pinton, 2008), resulting a variety of balanced and unbal-anced gametes (Danielak-Czech, Kozubska-Sobocińska, & Rejduch, 2016). For this reason, it is important to carry out cytogenetic screen-ing to select animals free of chromosomal aberrations that could easily reproduce without control, especially in the native breeds. The black Lucano pig (BLP) is an indigenous pig, ancient autochthonous genetic type (AAGT), typical of Basilicata region (Southern Italy). The breed Received: 6 February 2020 

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DOI: 10.1111/rda.13664 O R I G I N A L A R T I C L E

A de novo reciprocal chromosomal translocation t(3;6)(p14;q26)

in the black Lucano pig

Viviana Genualdo

_{| Cristina Rossetti}

_{| Alfredo Pauciullo}

_{| Petra Musilova}

_|

Domenico Incarnato

_{| Angela Perucatti}

Genualdo and Rossetti contributed equally to this work.

1_{Laboratory of Animal Cytogenetics and} Genomics, National Research Council (CNR), ISPAAM, Naples, Italy

2_{Department of Agricultural, Forest} and Food Sciences, University of Turin, Grugliasco, Italy

3_{Veterinary Research Institute, Brno, Czech} Republic

Correspondence

Alfredo Pauciullo, National Research Council (CNR), ISPAAM, Laboratory of Animal Cytogenetics and Genomics, Naples, Italy. Email: alfredo.pauciullo@unito.it

Abstract

In the past two decades, several cytogenetic screening programmes identified dif-ferent chromosome rearrangements in pig, most of which represented by reciprocal translocation (rcp). This chromosome abnormality does not involve the variation in the number of chromosomes, but only the rearrangement of genetic material, result-ing in phenotypically normal carriers with fertility problems. Durresult-ing an occasional cytogenetic screening, a new reciprocal translocation was detected in the black Lucano pig native breed. We analysed 15 animals reared by a family-run piggery in Basilicata region (Southern Italy). After karyotyping, four pigs (two boars and two sows) revealed two unpaired chromosomes. Analysis of the RBA karyotype and the dual-colour FISH technique confirmed that these pigs showed the same reciprocal translocation involving the chromosomes SSC3 and SSC6. The precise location of breakpoints was identified by RBH-FISH t(3;6)(p14;q26), whereas the analysis of the pedigree showed a case of Mendelian inheritance within a family, after the de novo occurrence of the new rcp. Considering the consequences of the rcp on the fertil-ity, this study points out the importance of the cytogenetic screening in the native breeds for the safeguard of the genetic biodiversity and the sustainability of the rural areas.

K E Y W O R D S

is included in the National Pedigree Register, where it is also known as Apulo-Calabrese breed (ANAS, 2020). According to FAO catego-ries described by Henson (1992), in 1997, the BLP population was in a critical status with only 22 pigs; therefore, an in situ conservation programme was started (DAD-IS, 2020). Nowadays, the BLP stock, reported by national farmer association of pig, consists of 509 sows, 80 boars, 2,086 and 1,891 young sows and boars respectively (ANAS, 2019). The BLP is particularly rustic, able to use scarce food resources and adapt to various types of environments, naturally raised in semi-wild pig grazing system and in non-cultivated areas. The BLP has a medium-small uniform blackish coat, elongated snout and long limbs, hard hooves and wild appearance. The females reach a weight of about 130 kg and males achieve about 150 kg. Peculiarities of this breed were found in qualitative and organoleptic traits of meat (Perna, Simonetti, Intaglietta, & Gambacorta, 2015a, 2015b). Indeed, a recent study ex-plained that native BLP meat might be considered a functional food since it is a good source of antioxidative and antihypertensive peptides (Simonetti, Gambacorta, & Perna, 2016).

To the best of our knowledge, the present study reports for the first time a reciprocal translocation in BLP breed, detected in four animals belonging to the same piggery. The cases were character-ised by classic banding analysis and fluorescence in situ hybridization (FISH) with chromosome-specific painting probe to precisely iden-tify the reciprocal translocation breakpoint.

2 | MATERIALS AND METHODS

2.1 | Ethical approval

Procedures were in accordance with the ethical standards of the na-tional ethics committee on research on animal science of 7th June 2011. All institutional and national guidelines for the care and use of laboratory animals were followed. The protocol was approved by the Committee on the Ethics of Animal experiments of the CNR-ISPAAM (Permit number 0001190 of 13th December 2018).

2.2 | Animals

Fifteen BLPs (nine boars and six sows) reared in the semi-wild state were sampled from a family-run piggery in Basilicata region (Southern Italy). Blood samples were collected by Vacutainer sodium-heparin tubes from all 15 BLPs. All subjects were in good physical condition and showed normal phenotype (Figure 1).

2.3 | Cell culture and banding techniques

Peripheral blood cultures were set up according to the standard protocols of our laboratory, as previously published (Perucatti et al., 2011). Two types of cell cultures were performed per each sample, normal cultures (without the addition of any analogue base) and

cultures treated for the late incorporation of 5-bromodeoxyurdine (BrdU) to obtain R-banding. The first were used for CBA-banding technique (C-banding by using acridine orange staining) and con-ventional dual-colour FISH, while the latter were used for both RBA-banding (R-banded karyotypes) and FISH mapping. CBA- and RBA-banding, as well as the sequential RBA/CBA techniques fol-lowed the protocol of Pauciullo et al. (2016).

2.4 | Probe production and fluorescent in situ

hybridization

Painting probes corresponding to the chromosomes SSC3 and SSC6 were produced by laser microdissection from GTG-banded meta-phases of a karyological normal domestic pig (Sus scrofa domestica) according to Kubickova, Cernohorska, Musilova, and Rubes (2002). Microdissected chromosomes were amplified by a DOP-PCR re-action and then labelled with Spectrum Green-dUTP (SSC3) or Spectrum Orange-dUTP (SSC6) in a secondary PCR. The probes were used for FISH. About 10 µl of the hybridization mixture (50% formamide, 10% dextran sulphate, 2× SSC) contained 5 µg of salmon sperm DNA (Sigma-Aldrich), 1.5 µg of pig competitor DNA (Applied Genetics Laboratories) and 0.1 µg of the probe.

Two fluorescent in situ hybridization (on conventional and RBH-band metaphases) were performed according to Pauciullo, Perucatti, Cosenza, et al. (2014) and Pauciullo, Perucatti, Iannuzzi, et al. (2014). The probes were denatured for 10 min at 72°C and pre-hybridized for 60 min at 37°C. Metaphase spreads were dena-tured for 3 min in a solution of 70% formamide in 2× SSC (pH 7.0) at 72°C. Denaturation was stopped in a scale 70%, 80% and 96% of cold ethanol and air-dried. The hybridization mixture was applied to the slides, covered with 24 × 24 mm coverslips and incubated in

a moist chamber at 37°C over-night. After hybridization, coverslips were removed by a gentle washing step in 2× SSC. The slides were then washed 3 × 4 min in a washing solution (50% formamide in 2× SSC) at 42°C, followed by three additional washing steps for 4 min in 2× SSC at 42°C and a further step for 5 min in PBST at room temperature. Finally, slides were counterstained with DAPI (4,6-di-amidino-2-phenylindole) solution (0.24 µg/ml; Sigma-Aldrich) in Antifade (Vector Laboratories).

2.5 | Microscopic analysis

By Leica DM2000 LED fluorescence microscope equipped with 100× oil immersion lens, FITC-, TXR- and DAPI-specific filters and camera. At least 100 cells were analysed for each animal with CBA-banding, to check the frequency of sexual chromosomes, while 20 cells treated for R-banding were karyotyped followed the pig stand-ard karyotype (Gustavsson, 1988b) and constructed using cytovision

7.4 software. Furthermore, at least 30 metaphases were studied for the sequential RBA/CBA technique.

3 | RESULTS AND DISCUSSION

All animals used in this study were karyotyped by RBA-banding. Eleven out of 15 BLPs showed a normal arrangement (2n = 38,XX or XY), whereas four animals (two sows and two boars) showed an exchange of genetic material between two non-homologous

chromosomes. Therefore, they were identified as carrier of the same balanced reciprocal translocation 2n = 38,XX or XY t(3;6). In particular, the karyotype pointed out the break at the level of p-arm of the SSC3 chromosome and q-arm of SSC6 (Figure 2a,b). The translocated animals showed a normal phenotype, and no re-productive problems were recorded by the farmer. Furthermore, the CBA-banding and the sequential RBA/CBA-banding did not show constitutive heterochromatin anomaly in both autosomes and sex chromosomes (Figure S1).

Classical cytogenetic techniques are still very useful to assess genetic defects, including reciprocal translocations. However, the variability of chromosome preparations, and limited banding reso-lution, makes the clarification of certain genetic events difficult, in-cluding the precise localization of rcp breakpoints. In this respect, the molecular methods show a higher resolution level and their combination with classic methods are more accurate (Pauciullo, Fleck, Lühken, Berardino, & Erhardt, 2013). In light of these consid-erations, a dual-colour FISH on conventional metaphases and the use of species-specific painting probes confirmed the chromosome involved in the translocation (Figure 3a). Furthermore, since the DAPI banding does not allow clear chromosome identification, we set up a RBH-FISH experiment to determine the position of break-points more accurately. With this method, it is possible, in fact, to visualize simultaneous R-banding (by late BrdU-incorporation and Hoechst staining) and fluorescein signals (by specific SSC3 painting probe) providing immediate and clearer results. According to the standard idiogram, the breakpoint on SSC3 was located in the p13-14 band and resulted in the translocation of the little portion of the

F I G U R E 2   Classical and molecular cytogenetics for the detection of the t(3;6)(p14;q26). (a) RBA-stained metaphase plate showing both SSC3, der3, SSC6 and der6. (b) Karyotype arranged by the metaphase of the Figure 2a. (c) RBH-FISH obtained by using specific SSC3 painting probe on pig metaphases shows specific signals on the normal chromosome and on the translocated der3 and der6. Fluorescein isothiocyanate signals were superimposed on RBH-banding (R-banding using early BrdU-incorporation and Hoechst staining). (d) Details of the RBH-FISH and diagrammatic representation of the SSC chromosomes involved in the reciprocal translocation t(3;6)(p14;q26) with the breakpoints (dotted line). FISH, fluorescence in situ hybridization

(a) (b)

SSC3p14-p17 arm onto the SSC6 chromosome q25 band, while the derivative chromosome (der) of the SSC6q26-q35 was translocated on the SSC3p13 chromosome (Figure 2c,d).

The only balanced reciprocal translocation described from a cy-togenetic point of view and involving the same chromosomes (SSC3 and SSC6) was reported by Villagomez and Pinton (2008). However, in that study, different chromosome breakpoints were found [t(3;6) (p14;q21)]. Despite an extensive analysis of the meiotic synaptone-mal complexes, the same authors refer to a finer molecular approach using DNA specific probes to confirm their observation.

In general, the chromosome 6 appears to be particularly prone to be involved in reciprocal translocation, resulting within the high-est tertile for the number of observed breakpoints (Donaldson et al., 2019). However, the presence of these chromosomal hotspots seems to be distributed in a non-random way and associated with the length of chromosomal band, chromatin density and presence of fragile sites (Donaldson et al., 2019; Riggs, Kuczek, Chrisman, & Bidwell, 1993; Rønne, 1995). In this respect, both SSC3 and SSC6 are characterised by common fragile sites (Rønne, 1995) that fall at the same band level (3p14) of the reciprocal translocation observed in the present study for SSC3, and in a very close proximity (6q28) for SSC6.

Excluding these specific examples involving SSC3 and SSC6, the prevalence of balanced structural chromosomal rearrangements in pigs was assessed between 0.5% and 1.5% depending on the inten-sity of cytogenetic screening within each population, and almost the totality of chromosomal aberrations found were reciprocal translo-cations (Ducos et al., 2007, 2008; Quach et al., 2016), also involving a sex chromosome t(Y;14)(q10;q11) in a single case of azoospermic boar identified by Pinton et al. (2008).

The reciprocal chromosome translocations are associated to a reduced litter size caused by lethal embryonic aneuploidy (Ducos, Berland, et al., 1998; Ducos, Pinton, et al., 1998; Gustavsson, 1980, 1988a; King, 1980; Pinton et al., 2000), reduction of the fertility and malformations not always manifested (Sánchez-Sánchez, Gómez-Fidalgo, Pérez-Garnelo, Martín-Lluch, & De la Cruz-Vigo, 2019). Although in most cases the carriers have no phenotypic disorders,

they have problems at the gametogenesis for the formation of a quadrivalent, a structure made of two normal and two rcp chromo-somes that can segregate in alternate way or in adjacent ways (type I or II). In the first case, gametes may result normal or carrying the bal-anced translocation; in the latter cases, they carry derivative chro-mosomes, are unbalanced and will generate segmental aneuploidy in the offsprings.

For avoiding reproductive problems or even infertility of the breeds, carriers are normally culled. For endangered breeds, where cytogenetic control does not exist, the presence of translocation carriers could be detrimental and carriers could spread quickly into the population, if not eliminated. However, in small breeds, the real occurrence might be different. Breeders might prefer to continue using the carrier subjects and karyotype the descendants with at least two aims. The first is to transmit the genetic value without the anomaly, and the second is to prevent an increase in consanguinity as in these breeds often the number of animal suitable for reproduc-tion is very limited.

Translocations detected in affected animals may be inherited from a parent who carry the balanced form of the rearrangement or may arise de novo. In this study, the pedigree analysis of the BLPs showed both situations (Figure 3b). In fact, the great-grandparents (first generation) were karyological normal, whereas the balanced rearrangement appeared de novo at the second generation with the grand-mother. The mating of this sow with two different boars gave at the third generation two piglets (a male and a female) with the same balanced rcp. Successively, the mating of the latter sow with a normal boar, gave at the fourth generation, a male piglet still carrying the same rcp. Therefore, this is a typical example of Mendelian in-herited balanced rcp within a family. Despite the chromosomal rear-rangement, all four BLPs carrying the rcp did not show any diversity compared with the characteristic of the breed, as confirmed by the ante- and post-mortem inspection (Figure S2).

In conclusion, reciprocal translocations usually induce a high per cent of genetically unbalanced gametes leading to an early mortality of the embryos produced. This implicates large negative F I G U R E 3   Dual-colour FISH and pedigree of the BLPs carriers of the t(3;6)(p14;q26). (a) The painting probe for SSC3 hybridizes on SSC3, the proximal part of der3 and the telomeric region of der6 (green signals). The painting probe for SSC6 hybridizes on SSC6, the proximal part of der6 and the distal part of der3 (red signals). (b) The pedigree of the reciprocal translocated BLPs shows the Mendelian inheritance within the family. Squares are boars; circles are sows. Empty symbols are normal; half-blacks are carriers of the balanced reciprocal translocation. BLP, black Lucano pig; FISH, fluorescence in situ hybridization

effects on productivity, produces management problems and in-come losses at the farm level. The identification of a new rcp in pigs confirms the importance of the cytogenetic screening as es-sential tool both for a genetic prophylaxis of commercial breeds and, in the specific case, for the safeguard and the protection of autochthonous/native genetic type (AGT) biodiversity like the Black Lucano Pig. In this respect, structuring a network among the associations involved in the protection of local breeds might be useful as model to establish national cytogenetic screening for the AGT worldwide and to sustain the rural areas by the enhancement and preservation of the traditional breeding and the related typi-cal food products.

ACKNOWLEDGEMENTS

The authors wish to thank Professor Leopoldo Iannuzzi (CNR-ISPAAM) for his invaluable scientific experience and advices, Mr. Giuseppe Grazioli and Giuseppe Auriemma (CNR-ISPAAM) for the technical support and Dr Domenico Mecca as breeder of BLPs. CONFLIC T OF INTEREST

None of the authors have any conflict of interest to declare. AUTHOR CONTRIBUTIONS

Viviana Genualdo and Cristina Rossetti contributed equally to conceptualization, methodology, investigation and data curation. Alfredo Pauciullo contributed equally to supervision, validation, writing-reviewing and editing. Petra Musilova contributed equally to investigation, validation and reviewing. Domenico Incarnato con-tributed equally to investigation and data curation. Angela Perucatti contributed equally to conceptualization, data curation, software and writing-original draft preparation.

DATA AVAIL ABILIT Y

All data supporting results presented herein are reported within this manuscript. No additional data archiving is necessary.

ORCID

Viviana Genualdo https://orcid.org/0000-0001-7635-4508

Cristina Rossetti https://orcid.org/0000-0003-1944-0068

Alfredo Pauciullo https://orcid.org/0000-0002-3140-9373

Petra Musilova https://orcid.org/0000-0001-7793-1960

Domenico Incarnato https://orcid.org/0000-0003-4945-986X

Angela Perucatti https://orcid.org/0000-0003-4642-9921

REFERENCES

ANAS (2019, June 28). Associazione Nazionale Allevatori Suini. Relazione

attività, parte terza, libro genealogico. Retrieved from http://www.

anas.it/circo lari/20190 0000.PDF

ANAS (2020, February 23). Associazione Nazionale Allevatori Suini,

Apulo-Calabrese breed. Retrieved from http://www.anas.it/docum enti/ Scheda_apulo calab rese.pdf

DAD-IS (2020, February 23). Domestic animal diversity information

sys-tem. Retrieved from http://www.fao.org/dad-is/brows e-by-count ry-and-speci es/en/

Danielak-Czech, B., Kozubska-Sobocińska, A., & Rejduch, B. (2016). Molecular cytogenetics in the diagnostics of balanced chromosome mutations in the pig (Sus scrofa)–A review. Annals of Animal Science,

16(3), 679–699.

Donaldson, B., Villagomez, D. A., Revay, T., Rezaei, S., & King, W. A. (2019). Non-random distribution of reciprocal translocation break-points in the pig genome. Genes, 10(10), 769.

Ducos, A., Berland, H.-M., Bonnet, N., Calgaro, A., Billoux, S., Mary, N., … Pinton, A. (2007). Chromosomal control of pig populations in France: 2002–2006 survey. Genetics Selection Evolution, 39(5), 583.

Ducos, A., Berland, H. M., Pinton, A., Guillemot, E., Seguela, A., Blanc, M. F., … Darre, R. (1998). Nine new cases of reciprocal translocation in the domestic pig (Sus scrofa domestica L.). Journal of Heredity, 89(2), 136–142.

Ducos, A., Pinton, A., Berland, H. M., Seguela, A., Blanc, M. F., Darre, A., & Darre, R. (1998). Five new cases of reciprocal translocation in the domestic pig. Hereditas, 128(3), 221–229.

Ducos, A., Revay, T., Kovacs, A., Hidas, A., Pinton, A., Bonnet-Garnier, A., … Iannuzzi, L. (2008). Cytogenetic screening of livestock populations in Europe: An overview. Cytogenetic and Genome Research, 120(1–2), 26–41.

Genualdo, V., Perucatti, A., Marletta, D., Castiglioni, B., Bordonaro, S., Iannaccone, M., … Iannuzzi, A. (2018). Cytogenetic investigation in two endangered pig breeds raised in Southern-Italy: Clinical and en-vironmental aspects. Livestock Science, 216, 36–43.

Gustavsson, I. (1988a). Reciprocal translocation in four boars producing decreased litter size. Hereditas, 109(2), 159–168.

Gustavsson, I. (1988b). Standard karyotype of the domestic pig: Committee for the standardized karyotype of the domestic pig.

Hereditas, 109(2), 151–157.

Gustavsson, I. (1980). Chromosome aberrations and their influence on the reproductive performance of domestic animals - A review.

Zeitschrift fur Tierzuchtung und Zuchtungsbiologie, 97(3), 176–195.

Henson, E. L. (1992). In situ conservation of livestock and poultry. Rome, Italy: FAO-ed.

King, W. A. (1980). Spontaneous chromosome translocations in cattle

and pigs: a study of the causes of their fertility reducing effect. PhD

Dissertation. Uppsala, Sweden: Swedish University of Agricultural Sciences.

Kociucka, B., Szczerbal, I., Bugaj, S., Orsztynowicz, M., & Switonski, M. (2014). A high incidence of adjacent-1 meiotic segregation pat-tern, revealed by multicolor sperm FISH, in a carrier boar of a new reciprocal translocation t (6; 16)(p13; q23). Cytogenetic and Genome

Research, 142(1), 21–27.

Kubickova, S., Cernohorska, H., Musilova, P., & Rubes, J. (2002). The use of laser microdissection for the preparation of chromosome-spe-cific painting probes in farm animals. Chromosome Research, 10(7), 571–577.

Pauciullo, A., Fleck, K., Lühken, G., Di Berardino, D., & Erhardt, G. (2013). Dual-color high-resolution fiber-FISH analysis on lethal white syn-drome carriers in sheep. Cytogenetic and Genome Research, 140(1), 46–54.

Pauciullo, A., Knorr, C., Perucatti, A., Iannuzzi, A., Iannuzzi, L., & Erhardt, G. (2016). Characterization of a very rare case of living ewe-buck hybrid using classical and molecular cytogenetics. Scientific Reports, 6, 34781. Pauciullo, A., Perucatti, A., Cosenza, G., Iannuzzi, A., Incarnato, D.,

Genualdo, V., … Iannuzzi, L. (2014). Sequential cross-species chro-mosome painting among river buffalo, cattle, sheep and goat: A use-ful tool for chromosome abnormalities diagnosis within the family Bovidae. PLoS ONE, 9(10), e110297.

Pauciullo, A., Perucatti, A., Iannuzzi, A., Incarnato, D., Genualdo, V., Di Berardino, D., & Iannuzzi, L. (2014). Development of a sequential multicolor-FISH approach with 13 chromosome-specific painting probes for the rapid identification of river buffalo (Bubalus bubalis, 2n= 50) chromosomes. Journal of Applied Genetics, 55(3), 397–401.

Perna, A., Simonetti, A., Intaglietta, I., & Gambacorta, E. (2015a). Fatty acids composition, cholesterol and vitamin E contents of Longissimus dorsi and Semitendinosus muscles of Suino Nero Lucano pigs slaugh-tered at two different weights. Animal Production Science, 55(8), 1037–1043.

Perna, A., Simonetti, A., Intaglietta, I., & Gambacorta, E. (2015b). Quality and sensory characteristics of culatello dry-cured products obtained from the Italian autochthonous pig Suino Nero Lucano and from a modern crossbred pig. Animal Production Science, 55(9), 1192–1199. Perucatti, A., Genualdo, V., Iannuzzi, A., De Lorenzi, L., Matassino, D.,

Parma, P., … Di Meo, G. P. (2011). A new and unusual reciprocal trans-location in cattle: Rcp (11; 25)(q11; q14–21). Cytogenetic and Genome

Research, 134(2), 96–100.

Pinton, A., Ducos, A., Berland, H., Seguela, A., Brun-Baronnat, C., Darré, A., … Yerle, M. (2000). Chromosomal abnormalities in hypoprolific boars. Hereditas, 132(1), 55–62.

Pinton, A., Raymond Letron, I., Berland, H. M., Bonnet, N., Calgaro, A., Garnier-Bonnet, A., … Ducos, A. (2008). Meiotic studies in an azo-ospermic boar carrying a Y; 14 translocation. Cytogenetic and Genome

Research, 120(1–2), 106–111.

Quach, A. T., Revay, T., Villagomez, D. A. F., Macedo, M. P., Sullivan, A., Maignel, L., … King, W. A. (2016). Prevalence and consequences of chromosomal abnormalities in Canadian commercial swine herds.

Genetics Selection Evolution, 48(1), 66.

Raudsepp, T., & Chowdhary, B. (2011). Cytogenetics and chromosome maps. In M. P. Rothschild & A. Ruvinsky (Eds.), The genetics of the pig (2nd Ed., pp. 134–178). Wallingford, CT: CAB International.

Riggs, P., Kuczek, T., Chrisman, C., & Bidwell, C. (1993). Analysis of aphid-icolin-induced chromosome fragility in the domestic pig (Sus scrofa).

Cytogenetic and Genome Research, 62(2–3), 110–116.

Rønne, M. (1995). Localization of fragile sites in the karyotype of Sus

scrofa domestica: Present status. Hereditas, 122(2), 153–162.

Rothschild, M. F., & Ruvinsky, A. (2011). The genetics of the pig. Wallingford, CT: CABI.

Sánchez-Sánchez, R., Gómez-Fidalgo, E., Pérez-Garnelo, S., Martín-Lluch, M., & De la Cruz-Vigo, P. (2019). Prevalence of chromosomal aberra-tions in breeding pigs in Spain. Reproduction in Domestic Animals, 54, 98–101.

Simonetti, A., Gambacorta, E., & Perna, A. (2016). Antioxidative and an-tihypertensive activities of pig meat before and after cooking and in vitro gastrointestinal digestion: Comparison between Italian autoch-thonous pig Suino Nero Lucano and a modern crossbred pig. Food

Chemistry, 212, 590–595.

Szczerbal, I., & Switonski, M. (2016). Chromosome abnormalities in do-mestic animals as causes of disorders of sex development or impaired fertility. In R. Payan-Carreira (Ed.), Insights from animal reproduction (pp. 207–225). IntechOpen. https://www.intec hopen.com/books / insig hts-from-anima l-repro duction

Villagomez, D., & Pinton, A. (2008). Chromosomal abnormalities, meiotic behavior and fertility in domestic animals. Cytogenetic and Genome

Research, 120(1–2), 69–80.

SUPPORTING INFORMATION

Additional supporting information may be found online in the Supporting Information section.

How to cite this article: Genualdo V, Rossetti C, Pauciullo A, Musilova P, Incarnato D, Perucatti A. A de novo reciprocal chromosomal translocation t(3;6)(p14;q26) in the black Lucano pig. Reprod Dom Anim. 2020;55:677–682. https://doi.