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1.
PLoS Pathog ; 20(8): e1012466, 2024 Aug.
Artículo en Inglés | MEDLINE | ID: mdl-39150989

RESUMEN

Most viral diseases display a variable clinical outcome due to differences in virus strain virulence and/or individual host susceptibility to infection. Understanding the biological mechanisms differentiating a viral infection displaying severe clinical manifestations from its milder forms can provide the intellectual framework toward therapies and early prognostic markers. This is especially true in arbovirus infections, where most clinical cases are present as mild febrile illness. Here, we used a naturally occurring vector-borne viral disease of ruminants, bluetongue, as an experimental system to uncover the fundamental mechanisms of virus-host interactions resulting in distinct clinical outcomes. As with most viral diseases, clinical symptoms in bluetongue can vary dramatically. We reproduced experimentally distinct clinical forms of bluetongue infection in sheep using three bluetongue virus (BTV) strains (BTV-1IT2006, BTV-1IT2013 and BTV-8FRA2017). Infected animals displayed clinical signs varying from clinically unapparent, to mild and severe disease. We collected and integrated clinical, haematological, virological, and histopathological data resulting in the analyses of 332 individual parameters from each infected and uninfected control animal. We subsequently used machine learning to select the key viral and host processes associated with disease pathogenesis. We identified and experimentally validated five different fundamental processes affecting the severity of bluetongue: (i) virus load and replication in target organs, (ii) modulation of the host type-I IFN response, (iii) pro-inflammatory responses, (iv) vascular damage, and (v) immunosuppression. Overall, we showed that an agnostic machine learning approach can be used to prioritise the different pathogenetic mechanisms affecting the disease outcome of an arbovirus infection.


Asunto(s)
Infecciones por Arbovirus , Virus de la Lengua Azul , Lengua Azul , Lengua Azul/virología , Lengua Azul/patología , Animales , Ovinos , Virus de la Lengua Azul/patogenicidad , Infecciones por Arbovirus/virología , Infecciones por Arbovirus/patología , Índice de Severidad de la Enfermedad , Modelos Animales de Enfermedad
2.
Vet Pathol ; 61(5): 803-814, 2024 Sep.
Artículo en Inglés | MEDLINE | ID: mdl-38613413

RESUMEN

ORF virus (ORFV) causes contagious ecthyma ("ORF"), a disease of sheep and goats characterized by lesions ranging from vesicles and pustules to atypical papilloma-like and angiomatous lesions in the skin and mucosae. The authors investigated the molecular factors leading to the ORF-associated atypical tumor-like changes. Fifteen lambs, 15 kids, and an adult ram clinically affected by natural ORFV infection were enrolled in the study and examined by several methods. ORFV was detected by viral culture or real-time polymerase chain reaction (RT-PCR) in the lesioned tissues and in the blood of the clinically affected sheep and goats. Surprisingly, ORFV was also detected in the blood of healthy goats from an affected herd. Microscopically, they found a pseudo-papillomatous proliferation of the epithelium, while the dermis and lamina propria were expanded by a proliferating neovascular component that highly expressed the viral vascular endothelial growth factor (vVEGF) and its host receptor vascular endothelial growth factor receptor 2 (VEGFR2). Immunohistochemistry, immunofluorescence, and in situ hybridization for mRNA showed that epidermal growth factor receptor (EGFR) was expressed in the fibrovascular component, in the infiltrating CD163+ macrophages, and in the basal stratum of the epidermis. Confocal immunofluorescence microscopy demonstrated that CD163+ macrophages were associated with VEGF and VEGFR2. Finally, they found by quantitative RT-PCR the overexpression of the interleukin-6 and VEGFR2 genes in the lesioned tissues. These findings suggest that ORFV activates an inflammatory reaction characterized by CD163+ macrophages expressing EGFR and VEGFR2, which might play an oncogenic role through synergistic action with vVEGF signaling.


Asunto(s)
Ectima Contagioso , Receptores ErbB , Enfermedades de las Cabras , Cabras , Inflamación , Virus del Orf , Animales , Virus del Orf/genética , Virus del Orf/aislamiento & purificación , Ovinos , Ectima Contagioso/patología , Ectima Contagioso/virología , Enfermedades de las Cabras/virología , Enfermedades de las Cabras/patología , Inflamación/veterinaria , Inflamación/patología , Inflamación/virología , Receptores ErbB/metabolismo , Receptores ErbB/genética , Masculino , Receptor 2 de Factores de Crecimiento Endotelial Vascular/metabolismo , Receptor 2 de Factores de Crecimiento Endotelial Vascular/genética , Factor A de Crecimiento Endotelial Vascular/metabolismo , Factor A de Crecimiento Endotelial Vascular/genética , Femenino , Antígenos CD/metabolismo , Antígenos de Diferenciación Mielomonocítica/metabolismo , Macrófagos/virología , Macrófagos/patología , Enfermedades de las Ovejas/virología , Enfermedades de las Ovejas/patología , Receptores de Superficie Celular
3.
J Virol ; 92(19)2018 10 01.
Artículo en Inglés | MEDLINE | ID: mdl-30021901

RESUMEN

Arboviruses can cause a variety of clinical signs, including febrile illness, arthritis, encephalitis, and hemorrhagic fever. The recent Zika epidemic highlighted the possibility that arboviruses may also negatively affect the male reproductive tract. In this study, we focused on bluetongue virus (BTV), the causative agent of bluetongue and one of the major arboviruses of ruminants. We show that rams that recovered from bluetongue displayed signs of testicular degeneration and azoospermia up to 100 days after the initial infection. Importantly, testicular degeneration was induced in rams experimentally infected with either a high (BTV-1IT2006)- or a low (BTV-1IT2013)-virulence strain of BTV. Rams infected with the low-virulence BTV strain displayed testicular lesions in the absence of other major clinical signs. Testicular lesions in BTV-infected rams were due to viral replication in the endothelial cells of the peritubular areas of the testes, resulting in stimulation of a type I interferon response, reduction of testosterone biosynthesis by Leydig cells and destruction of Sertoli cells and the blood-testis barrier in more severe cases. Hence, BTV induces testicular degeneration and disruption of spermatogenesis by replicating solely in the endothelial cells of the peritubular areas unlike other gonadotropic viruses. This study shows that a naturally occurring arboviral disease can cause testicular degeneration and affect male fertility at least temporarily.IMPORTANCE During the recent Zika epidemic, it has become apparent that arboviruses could potentially cause reproductive health problems in male patients. Little is known regarding the effects that arboviruses have on the male reproductive tract. Here, we studied bluetongue virus (BTV), an arbovirus of ruminants, and its effects on the testes of rams. We show that BTV was able to induce testicular degeneration in naturally and experimentally infected rams. Testicular degeneration was caused by BTV replication in the endothelial cells of the peritubular area surrounding the seminiferous tubules (the functional unit of the testes) and was associated with a localized type I interferon response, destruction of the cells supporting the developing germinal cells (Sertoli cells), and reduction of testosterone synthesis. As a result of BTV infection, rams became azoospermic. This study highlights that problems in the male reproductive tract caused by arboviruses could be more common than previously thought.


Asunto(s)
Virus de la Lengua Azul/patogenicidad , Lengua Azul/complicaciones , Endotelio Vascular/patología , Infertilidad Masculina/etiología , Enfermedades de las Ovejas/etiología , Espermatogénesis , Testículo/patología , Animales , Lengua Azul/patología , Lengua Azul/virología , Endotelio Vascular/metabolismo , Endotelio Vascular/virología , Infertilidad Masculina/patología , Masculino , Ovinos , Enfermedades de las Ovejas/patología , Testículo/metabolismo , Testículo/virología , Testosterona/análisis , Virulencia , Replicación Viral
4.
Parasitol Res ; 117(6): 1985-1988, 2018 Jun.
Artículo en Inglés | MEDLINE | ID: mdl-29671047

RESUMEN

Herein, we examined the brain of adult ewes and lambs less than 30 days old which were found affected by neurological signs in a flock located in Sardinia (Italy). Gross anatomo-pathological examination of all brains of the animals revealed multiple linear reddish-yellow foci of necrotic purulent inflammation due to oncosphere migration. Histologically, we confirmed a multifocal pyo-granulomatous meningo-encephalitis both in ewes and in lambs, confirming acute coenurosis. Morphological examination and DNA sequencing identified the Taenia multiceps we isolated as Tm1 strain. This report describes for the first time a natural acute coenurosis infection in suckling lambs under 30 days of age.


Asunto(s)
Infecciones por Cestodos/veterinaria , Meningoencefalitis/diagnóstico , Meningoencefalitis/veterinaria , Enfermedades de las Ovejas/diagnóstico , Taenia/aislamiento & purificación , Animales , Encéfalo , Infecciones por Cestodos/diagnóstico , Infecciones por Cestodos/parasitología , Ciclooxigenasa 1/genética , Brotes de Enfermedades , Femenino , Italia/epidemiología , Meningoencefalitis/parasitología , Ovinos , Enfermedades de las Ovejas/epidemiología , Enfermedades de las Ovejas/parasitología , Taenia/genética
5.
J Virol ; 89(19): 10044-52, 2015 Oct.
Artículo en Inglés | MEDLINE | ID: mdl-26202249

RESUMEN

UNLABELLED: Thirty-seven goats carrying different prion protein genotypes (PRNP) were orally infected with a classical scrapie brain homogenate from wild-type (ARQ/ARQ) sheep and then mated to obtain 2 additional generations of offspring, which were kept in the same environment and allowed to be naturally exposed to scrapie. Occurrence of clinical or subclinical scrapie was observed in the experimentally infected goats (F0) and in only one (F1b) of the naturally exposed offspring groups. In both groups (F0 and F1b), goats carrying the R154H, H154H, R211Q, and P168Q-P240P dimorphisms died of scrapie after a longer incubation period than wild-type, G37V, Q168Q-P240P, and S240P goats. In contrast, D145D and Q222K goats were resistant to infection. The immunobiochemical signature of the scrapie isolate and its pathological aspects observed in the sheep donors were substantially maintained over 2 goat generations, i.e., after experimental and natural transmission. This demonstrates that the prion protein gene sequence, which is shared by sheep and goats, is more powerful than any possible but unknown species-related factors in determining scrapie phenotypes. With regard to genetics, our study confirms that the K222 mutation protects goats even against ovine scrapie isolates, and for the first time, a possible association of D145 mutation with scrapie resistance is shown. In addition, it is possible that the sole diverse frequencies of these genetic variants might, at least in part, shape the prevalence of scrapie among naturally exposed progenies in affected herds. IMPORTANCE: This study was aimed at investigating the genetic and pathological features characterizing sheep-to-goat transmission of scrapie. We show that in goats with different prion protein gene mutations, the K222 genetic variant is associated with scrapie resistance after natural and experimental exposure to ovine prion infectivity. In addition, we observed for the first time a protective effect of the D145 goat variant against scrapie. Importantly, our results demonstrate that the phenotypic characteristic of the wild-type sheep scrapie isolate is substantially preserved in goats carrying different susceptible PRNP gene variants, thus indicating that the prion protein gene sequence, which is shared by sheep and goats, plays a fundamental role in determining scrapie phenotypes.


Asunto(s)
Enfermedades de las Cabras/genética , Enfermedades de las Cabras/patología , Proteínas PrPSc/genética , Proteínas PrPSc/aislamiento & purificación , Scrapie/genética , Scrapie/patología , Animales , Encéfalo/metabolismo , Encéfalo/patología , Femenino , Estudios de Seguimiento , Predisposición Genética a la Enfermedad , Genotipo , Enfermedades de las Cabras/transmisión , Cabras , Especificidad del Huésped , Italia , Masculino , Mutación , Fenotipo , Polimorfismo Genético , Embarazo , Scrapie/transmisión , Oveja Doméstica
6.
Viruses ; 16(2)2024 01 30.
Artículo en Inglés | MEDLINE | ID: mdl-38399985

RESUMEN

Canine herpesvirus 1 (CaHV-1) infects dogs, causing neonatal death and ocular, neurological, respiratory, and reproductive problems in adults. Although CaHV-1 is widespread in canine populations, only four studies have focused on the CaHV-1 whole genome. In such context, two CaHV-1 strains from both the kidney and spleen of 20-day-old deceased French Bulldog puppies were recently isolated in Sardinia, Italy. The extracted viral DNA underwent whole-genome sequencing using the Illumina MiSeq platform. The Italian CaHV-1 genomes were nearly identical (>99%), shared the same tree branch, and clustered near the ELAL-1 (MW353125) and BTU-1 (KX828242) strains, enlarging the completely separated clade discussed by Lewin et al., in 2020. This study aims to provide new insights on the evolution of the CaHV-1, based on high-resolution whole-genome phylogenetic analysis, and on its clinicopathological characterization during a fatal outbreak in puppies.


Asunto(s)
Enfermedades de los Perros , Infecciones por Herpesviridae , Herpesvirus Cánido 1 , Animales , Perros , Herpesvirus Cánido 1/genética , Filogenia , ADN Viral/genética , ADN Viral/análisis
7.
Viruses ; 16(1)2024 01 21.
Artículo en Inglés | MEDLINE | ID: mdl-38275968

RESUMEN

Orf virus (ORFV) belongs to the genus Parapoxvirus (Poxviridae family). It is the causative agent of contagious ecthyma (CE) that is an economically detrimental disease affecting small ruminants globally. Contagious ecthyma outbreaks are usually reported in intensive breeding of sheep and goats but they have also been reported in wildlife species. Notably, ORFV can infect humans, leading to a zoonotic disease. This study aims to elucidate the global evolutionary history of ORFV genomes in sheep and goats, including the first genomes from Central America in the analyses. In comparison to the last study on ORFV whole genomes, the database now includes 11 more sheep and goat genomes, representing an increase of 42%. The analysis of such a broader database made it possible to obtain a fine molecular dating of the coalescent time for ORFV S and G genomes, further highlighting the genetic structuring between sheep and goat genomes and corroborating their emergence in the latter half of 20th century.


Asunto(s)
Ectima Contagioso , Virus del Orf , Humanos , Ovinos , Animales , Virus del Orf/genética , Ectima Contagioso/epidemiología , Cabras , Rumiantes , Evolución Biológica , Filogenia
8.
Viruses ; 14(7)2022 07 04.
Artículo en Inglés | MEDLINE | ID: mdl-35891452

RESUMEN

Orf virus (ORFV) is distributed worldwide and is the causative agent of contagious ecthyma that mainly occurs in sheep and goats. This disease was reported for the first time at the end of 18th century in Europe but very little is currently known about the temporal and geographic origins of this virus. In the present study, the use of new Italian whole genomes allowed for better inference on the evolutionary history of ORFV. In accordance with previous studies, two genome types (S and G) were described for infection of sheep and goats, respectively. These two well-differentiated groups of genomes originated for evolutive convergence in the late 1800s in two different areas of the world (Europe for S type and Asia for G type), but it was only in the early 1900s that the effective size of ORFV increased among hosts and the virus spread across the whole European continent. The Italian strains which were sequenced in the present study were isolated on the Mediterranean island of Sardinian and showed to be exclusive to this geographic area. One of them is likely representative of the early European forms of ORFV which infected sheep and became extinct about one century ago. Such an ancient Sardinian strain may have reached the island simple by chance, where it quickly adapted to the new habitat.


Asunto(s)
Ectima Contagioso , Virus del Orf , Animales , Cabras , Virus del Orf/genética , Filogenia , Ovinos , Secuenciación Completa del Genoma
9.
Vet Sci ; 9(8)2022 Aug 19.
Artículo en Inglés | MEDLINE | ID: mdl-36006357

RESUMEN

Using a multidisciplinary approach, this report describes a clinical case of malignant catarrhal fever (MCF) occurring in a calf, which shared the pasture with sheep on a farm located in the island of Sardinia (Italy). We confirmed the conventional clinico-histopathological features of MCF, as well was the presence of Ovine herpesvirus type 2 (OvHV-2) DNA in several tissues, employing histological and virological investigations. The phylogenetic analysis revealed that this Sardinian OvHV-2 strain is genetically similar to all the other Italian strains. By Real Time PCR examinations of blood samples collected across Sardinia's sheep population, which is considered the most important reservoir species, we discovered an OvHV-2 prevalence ranging from 20 to 30 percent. Despite the high prevalence of OvHV-2 in the Sardinian sheep population, clinical disease in bovine remains sporadic; further investigations are needed to understand the risk factors that regulate this epidemiological aspect.

10.
Life (Basel) ; 11(5)2021 May 04.
Artículo en Inglés | MEDLINE | ID: mdl-34064326

RESUMEN

Orf virus (ORFV) represents the causative agent of contagious ecthyma, clinically characterized by mild papular and pustular to severe proliferative lesions, mainly occurring in sheep and goats. In order to provide hints on the evolutionary history of this virus, we carried out a study aimed to assess the genetic variation of ORFV in Sardinia that hosts a large affected small ruminant population. We also found a high worldwide mutational viral evolutionary rate, which resulted, in turn, higher than the rate we detected for the strains isolated in Sardinia. In addition, a well-supported genetic divergence was found between the viral strains isolated from sheep and those from goats, but no relevant connection was evidenced between the severity of lesions produced by ORFV and specific polymorphic patterns in the two species of hosts. Such a finding suggests that ORFV infection-related lesions are not necessarily linked to the expression of one of the three genes here analyzed and could rather be the effect of the expression of other genes or rather represents a multifactorial character.

11.
Vet Ital ; 54(4): 349-353, 2018 12 31.
Artículo en Inglés | MEDLINE | ID: mdl-30681135

RESUMEN

This paper reports that Bluetongue virus serotype 1 (BTV-1) infected blood collected during the 2006 Sardinia (Italy) epidemic from a ewe with clinical disease and stored at ~ 5°C for 10 years, caused Bluetongue (BT)-like clinical disease and death when inoculated into a susceptible Sarda breed ram. Anatomo-histopathological examination and Real-Time Reverse Transcriptase PCR (Real-Time RT-PCR) confirmed the presence of BTV-1 in several tissues proving that the BTV-1 2006 isolate has maintained its infectivity and virulence.


Asunto(s)
Virus de la Lengua Azul/fisiología , Virus de la Lengua Azul/patogenicidad , Lengua Azul/sangre , Criopreservación , Animales , Sangre/virología , Lengua Azul/virología , Virus de la Lengua Azul/genética , Italia , Refrigeración , Estaciones del Año , Serogrupo , Ovinos , Virulencia
12.
Parasite ; 25: 25, 2018.
Artículo en Inglés | MEDLINE | ID: mdl-29727269

RESUMEN

Echinococcus granulosus sensu lato is a zoonotic agent with a life cycle consisting of definitive hosts (dogs and wild carnivores), and intermediate hosts (usually ungulates). Other animals and humans may accidentally ingest eggs and contract cystic echinococcosis, acting as aberrant hosts. A 3-year-old neutered female cat was brought to a veterinary practice in Sassari (Italy) with abdominal distension. Ultrasound showed multiple intraperitoneal vesicles, which on laparotomy were found to be metacestodes of E. granulosus. Videos of the extraction of cysts are provided. Phylogenetic analysis based on a fragment of the cytochrome oxidase subunit 1 (cox1) mitochondrial gene identified the isolate as E. granulosus sensu stricto genotype G1, the most common genotype circulating in Europe and the Mediterranean basin. This is the first case report of cystic echinococcosis in domestic cats from Italy.


Asunto(s)
Equinococosis/veterinaria , Echinococcus granulosus/aislamiento & purificación , Zoonosis/epidemiología , Abdomen/diagnóstico por imagen , Abdomen/parasitología , Animales , Gatos , Equinococosis/diagnóstico , Equinococosis/epidemiología , Equinococosis/parasitología , Echinococcus granulosus/genética , Complejo IV de Transporte de Electrones/genética , Femenino , Genotipo , Italia/epidemiología , Laparotomía , Filogenia , Ultrasonografía , Zoonosis/diagnóstico , Zoonosis/parasitología
13.
Acta Vet Scand ; 58(1): 37, 2016 Jun 07.
Artículo en Inglés | MEDLINE | ID: mdl-27267214

RESUMEN

BACKGROUND: Neuroaxonal dystrophy (NAD) is a neurodegenerative condition affecting humans and animals characterized by the widespread presence of swollen axons (spheroids). CASE PRESENTATION: Herein, we report the pathological findings in a case of neuroaxonal dystrophy in a Dachshund-cross puppy, which was euthanized because of a proprioceptive positioning deficits and irreversible ataxia of the hind limbs. Histologically, there was a bilaterally symmetric neuroaxonal dystrophy with eosinophilic axonal spheroids exclusively localized at the level of the ventral posterior lateral nucleus of the thalamus, medial lemniscus, gracilis nucleus, medial cuneatus nucleus in the brain as well as the gracilis and cuneatus fasciculi throughout the spinal cord. CONCLUSION: To the authors' knowledge, this is the first report of canine neuroaxonal dystrophy with this exclusive and specific localization only in the neuronal circuit implicated in the transmission of conscious proprioceptive information.


Asunto(s)
Enfermedades de los Perros/patología , Distrofias Neuroaxonales/veterinaria , Animales , Axones/patología , Encéfalo/patología , Enfermedades de los Perros/diagnóstico , Perros , Miembro Posterior/patología , Distrofias Neuroaxonales/diagnóstico , Distrofias Neuroaxonales/patología , Médula Espinal/patología
14.
J Vet Diagn Invest ; 28(4): 449-54, 2016 Jul.
Artículo en Inglés | MEDLINE | ID: mdl-27154317

RESUMEN

Herein we describe a glioblastoma partially occupying the telencephalic portion of the left cerebral hemisphere of a Sardinian (syn. Sarda) breed ewe. Microscopically, the mass consisted of a pleomorphic spindle-shaped cell component organized as bundles and numerous small areas of round cells displaying an oligodendroglioma-like aspect. A high number of mitotic figures, large areas of necrosis surrounded by pseudopalisading glial cells, and multiple foci of dystrophic mineralization were also observed. The neoplasm was highly vascularized with glomerular vascular proliferation. Immunohistochemically, neoplastic cells proved to be strongly positive for nestin, vimentin, and olig-2, whereas they were invariably negative for synaptophysin. Few neoplastic cells and reactive astrocytes, mainly located at the edge of necrotic foci, proved to be positive for glial fibrillary acidic protein, whereas glomerular vascular proliferation was clearly positive for factor VIII and vascular endothelial growth factor. Gene sequencing analysis demonstrated homozygous p53 tumor suppressor gene (TP53) point mutations in the DNA-binding domain located in exon 8. The presence of round cells immunoreactive for olig-2 demonstrated that this tumor is a glioblastoma with oligodendroglioma component. Our pathologic, immunohistochemical, and molecular findings largely overlap those previously reported in humans and dogs.


Asunto(s)
Neoplasias del Ventrículo Cerebral/veterinaria , Glioblastoma/veterinaria , Oligodendroglioma/veterinaria , Enfermedades de las Ovejas/diagnóstico , Animales , Neoplasias del Ventrículo Cerebral/diagnóstico , Neoplasias del Ventrículo Cerebral/patología , Femenino , Glioblastoma/diagnóstico , Glioblastoma/patología , Oligodendroglioma/diagnóstico , Oligodendroglioma/patología , Mutación Puntual , Análisis de Secuencia de ADN/veterinaria , Ovinos , Enfermedades de las Ovejas/patología , Proteína p53 Supresora de Tumor
15.
Vet Microbiol ; 177(3-4): 409-13, 2015 Jun 12.
Artículo en Inglés | MEDLINE | ID: mdl-25840469

RESUMEN

In this study, we examined in Sardinia the brain of 555 autochthonous sheep, 50 goats, and 4 mouflons which were found affected by neurological signs. We found 6 goats and one mouflon with meningoencephalitis caused by Cryptococcus sp. There was no evidence of cryptococcal infections in any of the examined sheep. MLST genotyping on Cryptococcus sp. isolates identified Cryptococcus gatti genotype AFLP4/VGI and Cryptococcus neoformans var. neoformans genotype AFLP2/VNIV. Phylogenetically, all Cryptococcus gattii isolates fell within the autochthonous animal, human and environmental Mediterranean isolate cluster, forming a distinct branch along with environmental strains from Alicante, in the southern Mediterranean coast of Spain.


Asunto(s)
Criptococosis/veterinaria , Cryptococcus gattii/genética , Cryptococcus neoformans/genética , Enfermedades de las Cabras/microbiología , Meningoencefalitis/veterinaria , Enfermedades de las Ovejas/microbiología , Animales , Encéfalo/microbiología , Encéfalo/patología , Secuencia de Consenso , Criptococosis/microbiología , Cryptococcus gattii/clasificación , Cryptococcus gattii/patogenicidad , Cryptococcus neoformans/clasificación , Cryptococcus neoformans/patogenicidad , ADN de Hongos/análisis , Genotipo , Cabras , Humanos , Italia , Meningoencefalitis/microbiología , Tipificación de Secuencias Multilocus , Filogenia , Alineación de Secuencia/veterinaria , Ovinos , Oveja Doméstica , España
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