2015, Articolo in rivista, ENG
Iannuzzi A.; Perucatti A.; Pauciullo A.; Genualdo V.; De Lorenzi L.; Parma P.; Iannuzzi L.
One of the goals of molecular cytogenetics applied to livestock is the extension of their genetic physical maps, especially of loci containing genes related to productions. In this study, a comparative fluorescence in situ (FISH) mapping of three genes related to fecundity of cattle, river buffalo, sheep and goat is reported using bovine BAC-clones taking in account the data available on the BovMap database and considering their physical position and the data obtained from banding experiments. The following three gene sequences were mapped: tumor necrosis factor-? (TNF), correlated to male fertility; signal transducer and activator of transcription 5A (STAT5A), important for its influence on milk production and reproduction activity; melatonin receptor 1A (MTNR1A) important for reproductive seasonality. BAC probes containing these gene sequences were assigned by FISH, for the first time, on RB-banded chromosomes of these four important bovids. TNF was assigned to BTA/CHI23q21-22, OAR20q21-22 and BBU 2p21-22; STAT5A was assigned to BTA/CHI19q17-21, OAR11q17-21 and BBU3p15-21; MTNR1A was assigned to BTA/CHI27q14-15, OAR26q14-15 and BBU1p21-22. The three loci were located in homoeologous chromosomes and chromosome bands, underling the high degree of chromosome homologies among Bovids and extending the cytogenetic maps of this economically important species.
2015, Articolo in rivista, ENG
Iannuzzi A, Genualdo V, Perucatti A, Pauciullo A, Varricchio G, Incarnato D, Matassino D, Iannuzzi L.
A newborn calf of the Agerolese cattle breed underwent clinical cytogenetic investigation because of hyperflexion of the forelimbs, red eyes and the inability to stand. Anamnesis revealed that the mother, phenotypically normal, carried a chromosomal aberration. The newborn died after 2 weeks, and no remarkable alterations were found by the veterinarian on postmortem examination. The mother was a carrier of a reciprocal balanced translocation rcp(11;25)(q11,q14~21) detected after a cytogenetic investigation in 2011; however, the analysis of the newborn revealed a different chromosomal aberration with partial trisomy of chromosome 25 and partial monosomy of chromosome 11. In fact, the results showed both chromosomes 25, one chromosome 11 and only one long derivative chromosome (der11). FISH analysis, performed using BAC clones, confirmed the chromosomes and their regions involved. Finally, both the localization of the breakpoints on band q11 (centromere) of chromosome 11 and band q14-21 of chromosome 25, and the complete loss of the der25 identified the aberration as an unbalanced translocation 60,XX,der(11)t(11;25)(q11;q14~21). A comparison with human chromosomes was also performed to search for similarities and possible genes involved in order to study their effects, thus extending the knowledge of these aberrations by case reports.
2015, Articolo in rivista, ENG
Genualdo V.; Perucatti A.; Pauciullo A.; Iannuzzi A.; Incarnato D.; Spagnuolo M.S.; Solinas N.; Bullitta S.; Iannuzzi L.
Over the last decades, an increase of pollutants of diverse origin (industrial, military, mining, etc.) was recorded in several areas of Sardinia Island. We report the results of a multidisciplinary and complementary study based on cytogenetic and physiological analyses. The data obtained show the effects of the environmental impact on six sheep flocks (Sardinian breed) grazing on natural pasturelands next to possible polluted areas and compared to three herds grazing in different areas far from those potentially contaminated and used as control. Sister chromatid exchange (SCE) test was used as cytogenetic test to analyze chromosomal damages and it was performed on peripheral blood samples collected from 129 adult sheep (age. >. 4. years) randomly selected from polluted (92 animals) and control (37 animals) areas. Two types of cell cultures were performed: without (normal cultures) and with the addition of 5-BrdU. SCE-mean values estimated over 35 cells counted for each animal were 8.65. ±. 3.40, 8.10. ±. 3.50, 8.05. ±. 3.08, 7.42. ±. 3.34, 9.28. ±. 3.56 and 8.38. ±. 3.29 in the exposed areas, whereas the average values were 7.86. ±. 3.31 in the control group. Significant increases (P. <. 0.01) of SCEs were found in three investigated areas of Southern Sardinia. Furthermore, sheep of the same flocks were characterized for blood redox homeostasis in order to define the potential targets of oxidative damage and to identify biomarkers of the extent of animal exposure to environmental contaminants. The plasma levels of Asc, Toc and Ret were found to be significantly lower (P. <. 0.001) in exposed sheep (I, II, IV and V) than in the control group. TAC as well as GPx and SOD activities were higher in control than in the exposed groups (P. <. 0.001). Finally, plasma levels of N-Tyr, PC, and LPO were significantly lower (P. <. 0.001) in the control group than in the exposed groups.
2015, Articolo in rivista, ENG
Alfredo Mazza, Prisco Piscitelli, Cosimo Neglia, Giulia Della Rosa and Leopoldo Iannuzzi
The region of Campania (particularly Naples and Caserta) has experienced an emergency in the waste management cycle during past years. Although the most critical phase has been overcome after the construction of the incineration plant in Acerra (an old-fashioned technology built up over a few months, whose impact on environment and health has not yet been assessed), most of the underlying problems have not been resolved. The illegal burning of wheels, plastics, textiles, and other industrial residuals, along with the detection of two thousand toxic substance dumping sites, still represents major concerns of environmental pollution and population health. This review summarizes the most relevant studies, which analyzed chemical contamination (primarily dioxins and polychlorinated biphenyls (PCBs)) of the air, soil, water, animals, and humans in Campania. In addition, we reviewed information on population health (i.e., mortality data, congenital malformations, and cancer incidence). Moving from a detailed mapping of (mostly illegal) waste dumping sites in Campania, we have focused on recent studies which have found: (a) high concentrations of dioxins (>=5.0 pg TEQ/g fat) in milk samples from sheep, cows, and river buffaloes; (b) remarkable contamination of dioxin and PCBs in human milk samples from those living in the Naples and Caserta areas (PCDDs+PCDFs and dioxin-like-PCBs (dl-PCBs) assessed at 16.6 pg TEQ/g of fat; range: 7.5-43 pg/g of fat); (c) potential age-adjusted standardized mortality rates associated with some specific cancer types; (d) a statistically significant association between exposure to illegal toxic waste dumping sites and cancer mortality, even after adjustment by socio-economic factors and other environmental indicators.
2015, Articolo in rivista, ENG
Pauciullo A., Küpper J., Brandt H., Donat K., Iannuzzi L., Erhardt G.
Mycobacterium avium subspecies paratuberculosis (MAP) is a pathogenic bacteria responsible of the lethal Johne's disease in cattle. So far, several GWA studies have been carried out to identify chromosomal regions highly associated with Johne's disease. The aim of this study was to investigate the genetic variability within a pool of 7 genes (LAMB1, DLD, WNT2, PRDM1,SOCS5, PTGER4 and IL10) indicated by former GWA/RNASeq studies as putatively associated with MAP infections and to achieve a confirmation study of association with paratuberculosis susceptibility in a population of 324 German Holstein cattle (162 cases: MAP-positive and 162 controls MAP-negative) using ELISA and faecal cultural tests. SNP validation and genotyping information are provided, quick methods for allelic discrimination were set up and transcription factor binding analyses were performed. The SNP rs43390642:G>T in the WNT2 promoter region is associated with paratuberculosis susceptibility (p=0.013), suggesting a protective role of the T allele (p=0.043, OR 0.50 [0.25-0.97]). The linkage disequilibrium with the DLD rs134692583:A>T might suggest a combined mechanism of action of these neighbor genes in the resistance to MAP infection, which is also supported by a significant effect showed by the haplotype DLDT/WNT2T (p=0.047). In silico analysis predicted rs43390642:G>T and rs134692583:A>T as essential parts of binding sites for the transcription factors GR and C/EBP?, GATA-1 respectively, hence suggesting a potential influence on WNT2 and DLD gene expression. This study confirms the region (UMD 3.1:50639460-51397892) on BTA 4 as involved in tolerance/resistance to Johne's disesase. In addition,this study clarifies the involvement of the investigated genes in MAP infection and it contributes to the understanding of genetic variability involved in Johne's disease susceptibility.
2014, Comunicazione in rivista (Letter - Letter to editor), ENG
A. Perucatti, V. Genualdo, A. Iannuzzi, M.A.Colonna, F. Giannico, D. Incarnato, G. Lubrano-Lavadera, L. Vonghia, A. Caputi-Jambrenghi, L.Iannuzzi
2014, Comunicazione in rivista (Letter - Letter to editor), ENG
A. Iannuzzi, A. Perucatti, A. Pauciullo, V. Genualdo, D. Incarnato, L. Pucciarelli, L. De Lorenzi, P. Parma, L. Iannuzzi
2014, Comunicazione in rivista (Letter - Letter to editor), ENG
Rubessa, M.; Pauciullo, A.; Peretti, V.; Iannuzzi, L.; Ramunno, L.; Di Berardino, D.
2014, Comunicazione in rivista (Letter - Letter to editor), ENG
A. Pauciullo, A. Perucatti, A. Iannuzzi, D. Incarnato, V. Genualdo, L. Pucciarelli, D. Di Berardino, L. Iannuzzi
2014, Comunicazione in rivista (Letter - Letter to editor), ENG
A. Iannuzzi, A. Perucatti, A. Pauciullo, V.Genualdo, D. Incarnato, L. Pucciarelli, L. De Lorenzi, G. Varricchio, D. Matassino, P. Parma, L.Iannuzzi
2014, Articolo in rivista, ENG
Alfredo Pauciullo*, Angela Perucatti, Gianfranco Cosenza, Alessandra Iannuzzi,Domenico Incarnato, Viviana Genualdo, Dino Di Berardino, Leopoldo Iannuzzi
The main goal of this study was to develop a comparative multi-colour Zoo-FISH on domestic ruminants metaphases using a combination of whole chromosome and sub-chromosomal painting probes obtained from the river buffalo species (Bubalus bubalis, 2n = 50,XY). A total of 13 DNA probes were obtained through chromosome microdissection and DOP-PCR amplification, labelled with two fluorochromes and sequentially hybridized on river buffalo, cattle (Bos taurus, 2n = 60,XY), sheep (Ovis aries, 2n = 54,XY) and goat (Capra hircus, 2n = 60,XY) metaphases. The same set of paintings were then hybridized on bovine secondary oocytes to test their potential use for aneuploidy detection during in vitro maturation. FISH showed excellent specificity on metaphases and interphase nuclei of all the investigated species. Eight pairs of chromosomes were simultaneously identified in buffalo, whereas the same set of probes covered 13 out 30 chromosome pairs in the bovine and goat karyotypes and 40% of the sheep karyotype (11 out of 27 chromosome pairs). This result allowed development of the first comparative M-FISH karyotype within the domestic ruminants. The molecular resolution of complex karyotypes by FISH is particularly useful for the small chromosomes, whose similarity in the banding patterns makes their identification very difficult. The M-FISH karyotype also represents a practical tool for structural and numerical chromosome abnormalities diagnosis. In this regard, the successful hybridization on bovine secondary oocytes confirmed the potential use of this set of probes for the simultaneous identification on the same germ cell of 12 chromosome aneuploidies. This is a fundamental result for monitoring the reproductive health of the domestic animals in relation to management errors and/or environmental hazards.
2014, Poster, ENG
Pauciullo A., Küpper J., Brandt H., Donat K., Iannuzzi L., Erhardt G.
2014, Poster, ENG
A. Perucatti, V. Genualdo, A. Iannuzzi, L. Pucciarelli, D. Incarnato, A. Pauciullo, D. Marletta, G. Scopino, B. Castiglioni, V. Peretti, L. Iannuzzi
2014, Poster, ENG
A. Perucatti, V. Genualdo, A. Iannuzzi, M.A. Colonna, F. Giannico, D. Incarnato, G. Lubrano-Lavadera, L. Vonghia, A. Caputi-Jambrenghi, L. Iannuzzi
2014, Poster, ENG
A. Iannuzzi, A. Perucatti, A. Pauciullo, V. Genualdo, D. Incarnato, L. Pucciarelli, L. De Lorenzi, P. Parma, L. Iannuzzi
2014, Articolo in rivista, ENG
L. Cigliano, M. Strazzullo, C. Rossetti, G. Grazioli, G. Auriemma, F. Sarubbi, C. Iannuzzi, L. Iannuzzi, M.S. Spagnuolo
The effect of the stage of lactation on blood redox homeostasis of bovine and buffalo cows was evaluated. The investigation was carried out on early lactating and mid-late lactating cows, reared in a farm located in Campania (southern Italy). Plasma concentration of ?-tocopherol and ascorbate, the total antioxidant capacity (TAC), glutathione peroxidase (GPx), and superoxide dismutase activities were higher (P < 0.01) in mid-late lactating cows, thus suggesting a higher consumption of antioxidants during early lactation. Plasma concentration of protein-bound carbonyls (PC) and nitrotyrosine (N-Tyr), and the level of lipid hydroperoxides (LPO) were higher (P < 0.01) in early lactating cows, thus suggesting that lipid peroxidation and peroxynitrite production are crucial in determining oxidative modifications in plasma. TAC was positively correlated with ascorbate concentration (P < 0.03), and negatively correlated with PC concentration (P < 0.002), and ascorbate was negatively correlated with PC (P < 0.03) in mid-late lactating group. These findings demonstrate that circulating ascorbate plays a major role in preventing protein modifications induced by carbonyls, and that ascorbate scavenging effect is impaired during early lactation. We calculated a protein oxidative stress index as the ratio (PC + N-Tyr)/TAC multiplied by 100, and we found that this parameter was higher (P < 0.0001) in early lactating cows. Therefore, it could be useful for assessing the extent of protein oxidative damage in relation to the whole antioxidant status. Further, we suggest that the LPO/GPx ratio multiplied by 100 might be used as lipid oxidative stress index in lactating cows. This index was higher (P < 0.0001) in early lactating cows, and might represent a standard parameter for evaluating the lipid damage depending on a deficiency of the enzymatic antioxidant defence. These parameters are proposed for a possible effective description of physiological changes associated with lactation.
2014, Articolo in rivista, ENG
Pauciullo A, Perucatti A, Iannuzzi A, Incarnato D, Genualdo V, Di Berardino D, Iannuzzi L.
The development of new molecular techniques (array CGH, M-FISH, SKY-FISH, etc.) has led to great advancements in the entire field of molecular cytogenetics. However, the application of these methods is still very limited in farm animals. In the present study, we report, for the first time, the production of 13 river buffalo (Bubalus bubalis, 2n = 50) chromosome-specific painting probes, generated via chromosome microdissection and the DOP-PCR procedure. A sequential multicolor-FISH approach is also proposed on the same slide for the rapid identification of river buffalo chromosome/arms, namely, 1p-1q, 2p-2q, 3p-3q, 4p-4q, 5p-5q, 18, X, and Y, using both conventional and late-replicating banded chromosome preparations counterstained by DAPI. The provided 'bank' of chromosome-specific painting probes is useful for any further cytogenetic investigation not only for the buffalo breeds, but also for other species of the family Bovidae, such as cattle, sheep, and goats, for chromosome abnormality diagnosis, and, more generally, for evolutionary studies.
2013, Abstract in atti di convegno, ENG
Pauciullo A.1, Cosenza G.2, Apicella E.2, Gallo D.2, Erhardt G.3, Iannuzzi L.1, Ramunno L.2
2013, Articolo in rivista, ENG
Leopoldo IANNUZZI
Cytogenetics represents one of the modern biotechnologies applied to the genetic improvement of livestock, including the water buffalo (Bubalus bubalis ), one of the most important economic species raised in the world, especially in the East Countries. Cytogenetics covers several aspects of the genetic improvement. In the present paper, an update on the evolutionary, clinical and molecular cytogenetics of the water buffalo is reported. Evoluzionary cytogenetics: Two main types of buffaloes are present in the world: the African buffalo (Syncerus caffer) and the Asiatic buffalo (Bubalus bubalis). The latter has two subspecies: the river type (2n=50) and the swamp type (2n=48). These two species diverged by a tandem fusion translocation involving river buffalo chromosomes 4 and 9 (telomere of 4p and centromere of 9). This fusion was accompanied by loss of the nucleolus organizer regions (NORs) present in river buffalo chromosome 4p, and large portions of constitutive heterochromatin (HC) present in river buffalo chromosome 9. River and swamp buffaloes are normally crossbreed, especially to increase milk production in swamp buffaloes. The hybrid (2n=49) could have a lower fertility due to unbalanced chromosome embryos. Further informations differentiating tribe bovinae (cattle and water buffalo) from the remaining ones belonging to bovidae family will be given. Clinical cytogenetics: The study of water buffalo karyotype is very important because several studies have demonstrated that about 20% of females with reproductive problems (lack of oestrus in fertility age or large delay in the return in oestrus) show sex chromosome abnormalities and almost of them have been found to be sterile for serious damages occurring in internal sex adducts. In Italy, in collaboration with the National Buffalo Breeder Association (ANASB), cytogenetic controls of bulls and males addressed to the reproduction, as well a s on some females with reproductive problems, have been performed to eliminate the carriers of chromosome abnormalities, adding economic value to river buffalo breeding. These studies are routinely performed by using both CBA- and RBA- banding techniques. Specific cases are also studied by fluorescence in situ hybridization (FISH) technique and specific molecular markers (generally bovine BAC-clones). Molecular cytogenetics: The availability of specific molecular markers containing coding sequences (generally bovine or ovine BAC-clones) and the use of the FISH-technique have opened the door to the molecular cytogenetics also for water buffalo. Indeed, cytogenetic maps with specific and detailed location of loci containing expressed coding (and no-coding) sequences along the chromosomes, have been performed for this species, although these maps are not particularly dense. However, they are useful: (a) to increase our knowledge on water buffalo genome by detailed physical description of mapped loci per single chromosome and chromosome band; (b) to study such chromosome abnormalities; (c) to anchor genetic maps (linkage, RH-maps) to specific chromosome regions; (d) to delineate the karyotype evolution of river buffalo versus cattle, sheep and related bovids, as well as between bovids and humans, especially to transfer useful information from human to animal genomes.
2013, Abstract in atti di convegno, ENG
PAUCIULLO A.1, PERUCATTI A.1, IANNUZZI A.1, INCARNATO D.1, GENUALDO V.1, PUCCIARELLI L.1, DI BERARDINO D.2, IANNUZZI L.1