Bilateral sertoli and interstitial cell tumours in abdominal testes of a goat with polled intersex syndrome (PIS).

An 8-year-old, mixed breed, polled goat was presented for evaluation of male-like behaviour. Clinical findings included clitoromegaly, a heavily muscled neck, pronounced beard, and erect dorsal guard hairs, which are phenotypic characteristics commonly observed in intersex animals. Transrectal ultrasonography revealed the presence of two abdominal masses caudolateral to the uterine horns. Serum concentration of estradiol was elevated. Genetic evaluation was compatible with polled intersex syndrome defined by an XX karyotype without a Y chromosome or SRY gene. Based on gross and histologic evaluation, the abdominal masses were determined to be intra-abdominal testes, each of which was effaced by Sertoli cell and interstitial (Leydig) cell tumours. The Sertoli cell tumours (SCTs) represented two unique histologic patterns. Regardless of pattern, neoplastic Sertoli cells were consistently lipid laden and positive for vimentin. Interstitial cell tumours (ICTs) were negative for vimentin. Clinical and histopathologic findings suggest that prolonged exposure to steroids secreted by neoplastic Sertoli cells contributed to virilization. In addition, results from immunohistochemistry indicated that vimentin may be a valuable immunodiagnostic tool for differentiation between interstitial and Sertoli cell tumours in goats.

Anatomical and genetic study of an ancient animal tooth showing brachyodont and hypsodont mixed taxonomical characteristics.

A non-human dental piece was found in a Roman Empire tomb dated the 3rd century A.C. in Zaragoza (Spain). The morphology of this piece showed mixed brachyodont (carnivores) and hypsodont (herbivores) characteristics. As a result, the taxonomical assignation of the piece was impossible. Therefore, a protocol based on the DNA sequence of the cytochrome c oxidase subunit 1 mitochondrial region (COI) was applied. For this purpose, a pair of primers able to amplify this region in a large variety of animals was designed. The results point to a species of the Genus Bos (Family Bovidae). This assignation was later confirmed by these quencing of a short fragment of the mitochondrial D-loop region. A complete morphological description of the tooth is presented together with the DNA sequence study and comparison protocol.

Haplotype analyses, mechanism and evolution of common double mutants in the human LDL receptor gene.

Familial hypercholesterolemia (FH), an autosomal dominant inherited disorder resulting in increased levels of circulating plasma low-density lipoprotein (LDL), tendon xanthomas and premature coronary artery disease (CAD), is caused by defects in the LDL receptor gene (LDLR). Three widespread LDLR alterations not causing FH (c.1061-8T>C, c.2177C>T and c.829G>A) and one mutation (c.12G>A) with narrow geographical distribution and thought to cause disease were investigated. In an attempt to improve knowledge on their origin, spread and possible selective effects, estimations of the ages of these variants (t generations) and haplotype analysis were performed by genotyping 86 healthy individuals and 98 FH patients in Spain for five LDLR SNPs: c.81T>C, c.1413G>A, c.1725C>T, c.1959T>C, and c.2232G>A; most patients carried two of these LDLR variants simultaneously. It was found that both the c.1061-8T>C (t = 54) and c.2177C>T alterations (t = 62) arose at about the same time (54 and 62 generations ago, respectively) in the CGCTG haplotype, while the c.12G>A mutation (t = 70) appeared in a CGCCG haplotype carrying an earlier c.829G>A alteration (t = 83). The estimated ages of selectively neutral alterations could explain their distribution by migrations. The origin of the c.12G>A mutation could be in the Iberian Peninsula; despite its estimated age, a low selective pressure could explain its conservation in Spain from where it could have spread to China and Mexico, since the sixteenth century through the Spanish/Portuguese colonial expeditions.

A case of ovine female XY sex reversal syndrome not related to anomalies in the sex-determining region Y (SRY).

The human XY gonadal sex reversal is a rare disease. Primary amenorrhea, lack of secondary sexual characteristics and the possible existence of streak gonads are the most common signs of this syndrome. Five cases of this syndrome have been described in the last 42 years in bovines. However, to the knowledge of the authors, no similar descriptions are available in sheep. In the present report, we are presenting the case of a 4 year old ewe 2n = 54,XY showing primary sterility and total absence of masculinization signs. The promoter and the complete coding segment of the sex determining region Y (SRY) were sequenced and found to be absolutely normal. The presence of the testis-specific protein gene (TSPY) was also verified by means of PCR. Necropsy findings include short vagina, lack of uterus and complete absence of masculine remains. Streak gonads concordant with previous descriptions in humans were also found.

Microsatellite markers for the analysis of genetic variability and relatedness in red-legged partridge (Alectoris rufa) farms in Spain.

The usefulness of several microsatellites in the management of Alectoris rufa restocking farms is evaluated in the present report. We analysed seven microsatellite loci in 147 randomly chosen individuals from four Spanish farms. Global F(IS) values were not significant. Slight genetic differentiation was only found between Farms 1 and 4. The global first and second parent exclusionary powers were 0.742 and 0.930, respectively. Microsatellite analysis can estimate the genetic relatedness between pairs of individuals by means of the Identity index. Since genealogies are unknown in most farms, pairwise Identity index values proved to be helpful in the management of matings, and the improvement of reproductive success and fitness of the descendants. Mean Identity index values showed that individuals within farms were not genetically more related than expected under random mating. Variance of the Identity index values suggest the absence of closed familial groups in these farms.

Scrapie resistance alleles are not associated with lower prolificity in Rasa Aragonesa sheep.

Scrapie is a prion disease characterised by the accumulation of the pathological associated form of cellular prion protein (PrP(SC)) in the central nervous system. Susceptibility to scrapie is associated with polymorphism in the ovine prion protein (PrP) gene. The European Union has implemented scrapie control programs, relying on selective breeding for scrapie resistance; the use of ARR-carrier and the exclusion of VRQ-carrier were recommended. In this study, 4323 individuals from Rasa Aragonesa Sheep breed were genotyped for the PrP gene and the individual estimated breeding values (EBV) for prolificity were calculated. Most represented PrP alleles do not work against prolificity. Only a significant association between VRQ/VRQ genotype and a lower EBV was observed (p = 0.027, eta2 = 0.002). Therefore, avoiding reproduction of VRQ/VRQ individuals would not cause negative effect regarding prolificity.

Population genetics and paternity in Rasa Aragonesa sheep using DNA fingerprinting.

DNA fingerprinting was used to study the population genetics and paternity of the Rasa Aragonesa sheep (Ovis aries). Using a combination of Hae III and the M13 derived probe pV47, the mean number of bands per individual (5.422 +/- 0.309), the background band sharing coefficient (0.347), the mean population frequency of alleles (0.192) and the mean heterozygosity for bands (0.893) were calculated for 45 individuals from eight different farms in Aragon, northern Spain. Therefore, the estimated probability of missing a wrong paternity was 0.126 for unrelated males, and this probability was 0.438 for full-sibs males. In addition, in a field trial of this technique, paternity was assigned for three ewe-lamb pairs with a probability of 97 per cent in two cases and 77 per cent in one case.

Analysis of microsatellites and paternity testing in Rasa Aragonesa sheep.

Four microsatellite loci (MAF50, MAF18, OarFCB20 and MCM527) were studied in Rasa Aragonesa sheep in order to evaluate their use in paternity testing. Several population characteristics were estimated [allele frequencies. effective allele number (Ne), polymorphism informative content (PIC) and probability of excluding wrong paternities (Pe)]. In 32 randomly chosen individuals, four alleles were detected for MAF50, with 2.55 effective alleles, 0.58 PIC and 0.35 Pe. For MAF18, five alleles were identified, with 2.99 effective alleles, 0.51 PIC and 0.32 Pe. For oarFCB20, 10 alleles were observed, with 6.06 effective alleles, 0.82 PIC and 0.68 Pe. Finally, for MCM527, six alleles were found, with 3.75 effective alleles, 0.69 PIC and 0.50 Pe. When these loci were used together with serum transferrin locus, Pe rose to 97.20 per cent. Field trials confirmed the real usefulness of these techniques.

Confirmation of assignment of the bovine K-casein locus by PCR.

The provision of a bovine gene map will allow the ready identification of genetic disease in cattle and will lead to the identification of the genetic loci responsible for quantitative traits of economic importance. An extension of the polymerase chain reaction to the identification of linkage in bovine-Chinese hamster cell hybrids has improved the speed and facility of the assignment of genes to linkage groups and thus makes it easier to achieve a bovine linkage map.

Canine stage 1 periodontal disease: a latent pathology.

To evaluate the potential health issues associated with periodontal disease (PD) in dogs, 1004 teeth from 25 dogs were examined. The dogs were randomly selected, aged 2-14 years, and had at least 95% of their teeth at the first PD stage. Significant positive correlations between plaque grade (PG) and gum inflammation, gingival regression, periodontal pocket, age and serum alanine aminotransferase (ALT) activity were identified. In contrast, PG was negatively correlated to total platelet count. Altogether, these findings suggest that prevention and therapy at the first PD stages can have an important impact on the general health condition of dogs.

Ovine heritable arthrogryposis multiplex congenita with probable lethal autosomal recessive mutation.

A novel congenital syndrome in lambs is described that is characterized by: (1) kyphoscoliosis (severe curvature, hunching and twisting of the thoracic spine, with associated abnormalities of the ribs and sternum), (2) distal arthrogryposis of the carpal and tarsal joints, and (3) cleft hard and soft palate or palatoschisis (a median fissure of the palate). Male and female lambs are affected equally and in multiple births there may be only one affected lamb. The affected lambs are born full term, but die shortly after birth because they are unable to start breathing. Affected lambs have slightly reduced bodyweight (due to low muscle mass) compared with normal newborn lambs of the same flock. The syndrome is remarkably similar to the recently described bovine heritable arthrogryposis multiplex congenita in the Angus breed. Genetic analysis of the flock by means of DNA microsatellite markers, together with study of the reproductive management of the flock, allowed us to discount an environmental aetiology (e.g. intoxications or infections). In contrast, evidence supports an autosomal recessive inherited disease for which the name ovine heritable arthrogryposis multiplex congenita is proposed.

Sry-negative XX true hermaphroditism in a roe deer.

A two-year-old roe deer was brought down in the course of a hunt in the north of Spain (Asturias). On physical examination the individual presented well-developed bared antlers, but surprisingly a female external genitalia. Several anatomical, histological and genetic analyses were performed in order to explain the observed phenotype. Necropsy evidenced ovary-like structures with follicles on the surface; histological analyses of testes evidenced positive immunolabel against testosterone in Leydig cells; genetic analyses showed that the sex of the individual was consistent with a female individual. PCR analysis failed to detect SRY sequences; no PIS deletion, which is responsible for XX sex-reversal in goats, was detected. On the basis of its presumptive normal female sexual karyotype (XX) and the presence of two functional abdominal bilateral testes and ovaries, the roe deer was finally diagnosed as possessing an XX hermaphroditism syndrome. However, as in many other cases, the specific reason for the occurrence of this case of hermaphroditism could not be determined.

Bilateral Leydig cell tumor in a six-year-old intersex goat affected by Polled Intersex Syndrome.

A 6-year-old, sterile, Blanca Celtibérica breed adult doe was referred to our faculty. The doe had external female genitalia, a short anogenital distance, and normally shaped udders. Masculinization signs in the head shape and male behavior were also noted at the time of referral. Genetic analysis demonstrated normal 2n = 60 XX karyotype and an absence of the sex-determining region Y (SRY). The animal was homozygous for a DNA deletion responsible for the Polled Intersex Syndrome (PIS). A uterus and 2 uterine horns were present at the postmortem examination. Gartner's ducts and degenerated Wolffian derivatives persisted. There were 2 intra-abdominal testicle-like structures, one of which consisted of epididymal and deferent ducts. An advanced Leydig cell tumor, resulting in almost total destruction of the intratesticular structures, was also observed. Leydig cell tumors usually produce testosterone. Thus, these histologic findings are compatible with the evident virilization.

Phylogenetic relationships among Spanish goats breeds.

We partially sequenced the mitochondrial hypervariable region 1 (HVR1) in 60 goats belonging to six Spanish breeds. The analysis of these and previously published sequences reveals a weak phylogeographical structure in the Iberian Peninsula breeds. Individuals from a single breed did not group into a single cluster. Furthermore, individuals from different breeds often shared single phylogenetic tree branches after UPGMA analysis. This could reflect the non-existence of breed isolation because of traditional seasonal pastoralism and annual long-distance migrations. Three goats belonging to the C maternal lineage were found, demonstrating a wider than previously thought distribution for this lineage.

Assignment of two markers carried by human chromosome 1 to different cattle synteny groups: FH to U1 and PEPC to U17 (chromosome 8).

Two loci located on human chromosome 1 were mapped in cattle by means of interspecific (hamster x cattle) somatic cell hybridization. FH is assigned to the U1 synteny group together with PGD, ENO1, AT3, and REN, while PEPC is found to belong to the U17 group (chromosome 8), like FN1, CRYG, and VIL1.

Cloning of the bovine activin receptor type II gene (ACVR2) and mapping to chromosome 2 (BTA2).

The cDNA for the bovine activin receptor type II (ACVR2) gene has been cloned and sequenced. It encodes a protein of 513 amino acids which is highly homologous (approximately 98% identity) to the human, mouse, and rat proteins. Using PCR analysis of bovine x hamster somatic cell lines, the ACVR2 gene was mapped to syntenic group U17, which has been localised to bovine chromosome 2. Comparative mapping has shown that the genes within U17 are also in a syntenic group on the long arm of human and sheep chromosome 2, as well as on mouse chromosome 1. This group of genes represents an evolutionarily conserved mammalian chromosomal segment. Genotyping a highly polymorphic microsatellite locus, (AT)4(GT)9(AT)11, found within an intron of this gene confirmed the localisation by linking the ACVR2-associated microsatellite to the region of chromosome 2 flanked by CSSM42 and TGLA226. This gene locus, which has the characteristics of a type I and type II mapping locus, represents the first localisation of this gene in any species to date.