Hair loss and endocrine dermatosis on horses raised in a Mimosa pudica var. unijuga-invaded area.

A Mimosa pudica var. unijuga-associated toxicity affecting horses occurred in Araguari, Triângulo Mineiro, Southeast Brazil. Affected horses had gradual hair loss of the mane and tail and endocrine dermatosis after grazing for three months during the dry season on a paddock invaded by the plant. The main histological lesions include compact ortho-keratotic hyperkeratosis and numerous flame follicles. Toxicological analysis by HPLC-UV demonstrated 0.8 mg/g of mimosine in the leaves.

A review on mycotoxins and mycotoxicoses in ruminants and Equidae in South America.

Of the mycotoxicoses caused by molds contaminating grains or their byproducts, leukoencephalomalacia of horses and less frequently aflatoxicosis in cattle have been reported in South America. However, the most important group of mycotoxins in the region are those caused by fungi that infect forages and other types of plants and have regional distribution. In this group, ergotism is important, both caused by Claviceps purpurea infecting grains or by Epichloë coenophiala infecting Schedonorus arundinaceus. Other important mycotoxicoses are those caused by indole-diterpenes produced by Clavicipitaceous fungi including Claviceps paspali in Paspalum spp., Claviceps cynodontes in Cynodon dactylon, and by Periglandula a seed transmitted symbiont associated with the tremorgenic plant Ipomoea asarifolia. The latter is an important poisoning in the northeastern and northern Brazil. Other important mycotoxicoses are those caused by swainsonine containing plants. It was demonstrated that swainsonine contained in Ipomoea carnea var. fistulosa is produced by an epibiotic fungus of the order Chaetothyriales whose mycelia develop on the adaxial surface of the leaves. Swainsonine is also produced by the symbiotic, endobiotic fungi Alternaria section Undifilum spp., which is associated with Astragalus spp. in the Argentinian Patagonia causing poisoning. Another form of mycotoxicosis occurs in poisoning by Baccharis spp., mainly B. coridifolia, a very important toxic plant in South America that contains macrocyclic trichothecenes probably produced by an endophytic fungus that has not yet been identified. Pithomycotoxicosis caused by Pithomyces chartarum used to be an important mycotoxicosis in the region, mainly in cattle grazing improved pastures of legumes and grasses. Slaframine poisoning, diplodiosis and poisoning by barley contaminated by Aspergillus clavatus has been rarely diagnosed in Brazil, Uruguay and Argentina.

Bilateral stringhalt associated with Hypochaeris radicata in Uruguayan horses.

We reported thirteen cases of bilateral stringhalt associated with Hypochaeris radicata that occurred in horses in Uruguay during a severe drought in the summer of 2023. All horses were affected chronically and progressively by bilateral hyperflexion of hindlimbs. In two severely affected horses, the main histological lesions included neuronal chromatolysis and axonal spheroids in the ventral gray horn in the lumbar and sacral spinal cord and axonal degeneration and digestion chambers in ventral roots fibers and long peripheral nerves. We suggest that in addition to injuries to peripheral nerves, lesions in the spinal cord play an important role in the clinical signs of stringhalt in horses.

Hepatic and renal lesions in sheep intoxicated with Urochloa hybrid Mulato II in Argentina.

A flock of 48 sheep in Argentina grazing on a pasture of hybrid Urochloa (formerly Brachiaria) Mulato II (Urochloa ruziziensis × Urochloa decumbens × Urochloa brizantha) developed facial dermatitis, severe jaundice, and weakness after brief physical activity. Blood biochemistry of 3 animals revealed azotemia, elevated aspartate aminotransferase activity, and increased direct, indirect, and total bilirubin concentrations. The urine was markedly turbid and contained large concentrations of bile pigments and protein. At autopsy of 2 animals, there was severe jaundice and subcutaneous submandibular edema. The livers were enlarged, intensely yellow, and had a marked acinar pattern. Gallbladders were distended, and the kidneys were diffusely dark in one animal and yellow-green in the other. Microscopically, there was lymphoplasmacytic and histiocytic cholangiohepatitis with abundant crystals in the lumen of bile ducts and in the cytoplasm of macrophages. The proximal and distal convoluted renal tubules had protein casts in their lumens, and crystals were observed in the lumen and epithelial cells. Lectin histochemistry showed strong affinity for Arachis hypogaea agglutinin in hepatic macrophages. In the one sheep that was tested for heavy metals, copper concentrations in the liver and kidney were within the RIs. Despite the immediate change of pasture, morbidity and mortality were 100% within 3 mo. The association between the consumption of this pasture, and the clinical, biochemical, pathology, and lectin histochemistry findings confirmed intoxication with Urochloa hybrid Mulato II. To our knowledge, intoxication by this hybrid of Urochloa has not been reported previously.

Leucaena leucocephala toxicity in Brazilian horses.

Leucaena leucocephala poisoning is reported in horses in different Brazilian regions. The poisoning occurred one month after the horses were introduced into paddocks invaded by the plant or after 10 days of consuming cut Leucaena administered as the only food. Affected horses showed moderate to severe hair loss on the mane and tail, orthokeratotic hyperkeratosis with marked follicular telogenization, and hyperplasia of thyroid follicular cells. Mimosin concentration in leaves (5.5 mg/g) was determined by a new HLPC-UV method which is also reported.

Cecal dilatation in cattle ingesting Megathyrsus maximus (syn. Panicum maximum) cv. Mombaça and cv. Tanzânia.

Nine outbreaks of cecal dilatation are reported in cattle in rotationally-grazed pastures of Megathyrsus maximus during the rainy season with one day grazing and rest intervals of 23-28 days. Clinical signs, observed 6-24 h after entering the pasture, were inappetence, long periods in sternal recumbency, rough hair coat, arched back, tenesmus with difficult defecation, dry stools, reduced ruminal movements, intestinal tympany with right abdominal distention, and decreased milk production. The animals recovered 8-12 h after their removal from the pastures. All pastures were fertilized at 80-120 kg/ha of urea/year. In six farms, in addition to urea, chicken litter (CL) was also added. Water-soluble carbohydrates (WSC), non-fiber carbohydrates (NCF) and starch concentrations in the pastures causing tympany were higher (P < 0.05) than in control pastures. To demonstrate the relationship of the disease with fertilization of the pastures, a pasture of M. maximus was divided into five blocks of 16 m2 and each block subdivided into four plots each, totaling 20 plots. One plot of each block was randomly chosen for 1 of 4 different treatments: 1. fertilized with 8 t/ha of CL; 2. fertilized with 16 t/ha of CL; 3. fertilized with 180 kg/ha urea; 4. control without fertilization. In general, plots fertilized with urea or with CL had higher NFC and starch concentrations (p < 0,05) than the control plots. In another experiment a pasture of M. maximus was divided into 23 paddocks and rotationally grazed. Ten paddocks were fertilized with 180 kg/ha of urea and five fertilized with 8 t/ha of CL. The concentration of NFC and starch were significantly higher (p < 0.05) in the pastures fertilized with urea and with urea plus CL. These results suggest that cecal dilatation may be caused by the high concentrations of NFC and starch in the pastures fertilized with urea and/or poultry litter.

The Occurrence of Cattle Tick Fever in a Region of the Atlantic Forest on the Border with the Caatinga in Brazil.

The Atlantic Forest biome is considered an area in which tick fever is endemic, while the Caatinga biome is considered an area in which tick fever is non-endemic. A survey on cattle tick fever was carried out in 448 cattle farms located in an area of the Atlantic Forest biome which borders the Caatinga biome. A retrospective study of five years, conducted from January 2018 to October 2022, allowed for the identification of the occurrence of tick fever in 44 out of the 448 herds visited. In 70.5% (31/44) of the herds involved, the disease was caused by Anaplasma marginale; in 6.8% (3/44), the disease was caused by Babesia spp.; and in 22.7% (10/44), the disease was caused by a coinfection of Babesia spp. and A. marginale. The disease mainly occurred from August to November (23/44). Bos taurus or B. taurus crossbreed animals were most affected (29/44) in an area in which 94% of the cattle were Bos indicus and 6% were B. taurus and B. taurus crossbreeds. In 24 herds (with adults affected in 17 and calves in 7), the implicated animals had recently been moved to the Atlantic Forest. In the other 20 herds (calves with heavy tick infestations were affected in 17 and adults in 3), tick fever occurred in non-moved cattle. Even though it is an endemic zone, tick fever is common and mainly affects B. taurus cattle, including adults moved from areas with enzootic instability and calves under nine months old with high tick loads.

Poisoning by Baccharis coridifolia in Early-Weaned Beef Calves: Pathological Study and New Macrocyclic Trichothecene Identification.

This study investigated two outbreaks of spontaneous poisoning by Baccharis coridifolia (Asteraceae) in early-weaned beef calves in Tacuarembó, Uruguay. A total of 34 affected calves showed signs of salivation, anorexia, apathy, marked dehydration, and diarrhea. Deaths occurred 36-72 h after consumption and mortality varied from 37.5% to 43.3% for outbreak 1 and outbreak 2, respectively. The main pathological findings include diffuse severe necrosis of the prestomachs and lymphoid tissues. Ultrastructurally, epithelial cells of the rumen showed swelling, lysis of the organelles, degradation of intercellular attachments, and degradation of the nuclear chromatin. Using LC-MS with diagnostic fragmentation filtering, 56 macrocyclic trichothecenes including glycosyl and malonyl conjugates were identified. The total concentration of macrocyclic trichothecenes, including conjugates, was estimated to be 1.2 ± 0.1 mg/g plant material. This is the first report of these malonyl-glucose conjugates from Baccharis coridifolia.

Lantana camara poisoning in cattle that took refuge during a storm in a forest invaded by this plant.

An outbreak of poisoning by Lantana camara occurred in cattle when a herd sought refuge in a Eucalyptus forest heavily infested by this plant. The animals showed apathy, elevated serum activities of hepatic enzymes, severe photosensitivity, jaundice, hepatomegaly and nephrosis. After a clinical manifestation period of 2-15 days, 74 out of 170 heifers died. The main histological changes were random hepatocellular necrosis, cholestasis, biliary proliferation and, in one animal, centrilobular necrosis. Immunostaining for Caspase 3 detected scattered apoptotic hepatocytes.

Epidemiological aspects of cattle tick fever in Brazil.

The present work reviews the epidemiologic situation of Anaplasma marginale and Babesia spp. infections and the occurrence of cattle tick fever outbreaks in Brazil. In areas of tick fever enzootic instability, environmental conditions interfere with the development of Rhipicephalus (Boophilus) microplus: chilly winter in the southern region, floods in the Pantanal, and low humidity in the Caatinga. In contrast, the climatic conditions of stable zones (Cerrado, Amazon and Atlantic Forest biomes) favor tick development. In enzootic areas, tick fever is uncommon because the animals are in frequent contact with the parasite, acquiring immunity naturally during the period of innate resistance; however, outbreaks may occur when calves become infested by considerable numbers of infected ticks during this period or in adults raised in tick-free environments that become infested for the first time when transporting to stable areas. It is necessary to better understand the disease's risk factors under stable conditions and the implications of the mechanical and other vector transmission of A. marginale. To prevent tick fever outbreaks in Brazil, it is important to develop and use anaplasmosis and babesiosis vaccines in cattle from enzootic unstable regions, especially when animals are moved to stable areas.

Plants causing poisoning outbreaks of livestock in South America: A review.

This paper reviews toxic plants for livestock in South America. We included 219 plants in this review, considering only the plants that caused at least one demonstrated outbreak of poisoning. Plants reported as toxic in other regions and present in South America, but with no confirmed outbreaks of poisoning in this region, are not included. We report the main aspects of the epidemiology, clinical signs, pathology and economical importance of the poisonings. Initially, the toxic plants are described based on their toxic compounds, but because the toxic compound is unknown in 92 (42%) of the plants we also classify them according to the affected system. This review highlights the great diversity of toxic plants capable of affecting livestock in South America, and for this reason studies in this regard should be intensified.

Toxic plants from the perspective of a "Quilombola" community in the Cerrado region of Brazil.

A multi-disciplinary team surveyed ranchers at the Kalunga Historical and Cultural Heritage Site, in the Cerrado region of west central Brazil, to determine impacts promoted by toxic plants on cattle. The expedition to the Kalunga region was carried out by Brazilian and American researchers. Previously selected cattle ranch properties from "Vão das Almas", "Engenho II" and "Vão do Moleque" were visited. Twenty-four interviews were carried out with cattle ranchers and a questionnaire was applied to obtain information about outbreaks of native plant poisoning and their effects on livestock, and the use of local plants in phytotherapy. We classified problematic plants into three distinct categories. First, the toxic plants most cited by residents causing cattle losses were the flowers of Caryocar brasiliense Cambess ("pequi"), the fruits of Terminalia corrugata (Ducke) Gere & Boatwr. (Buchenavia tomentosa Eichler - "mirindiba" or "pau-pilão"), Eugenia dysenterica (Mart.) DC ("cagaita"), and Palicourea marcgravii A. St. Hil ("erva-café" or "cafezinho"). Secondly, other plants considered toxic, but causing less severe losses were Emmotum nitens (Benth.) Miers ("casco d'anta"), Indigofera lespedezioides (Kunth) ("timbozinho"), Ricinus communis L. ("mamona"), Pteridium esculentum (G. Forst.) Cockayne ("samambaia"), Stryphnodendron adstringens (Mart.) Coville ("barbatimão"), and Actinocladum verticillatum (Nees) McClure ex Soderstr. ("cambaúba"). The most important finding was the identification of the C. brasiliense flower as potentially toxic to cattle, which must be subject for future research. Further, we confirmed the toxicity and importance of P. marcgravii, E. dysenterica, and Terminalia corrugata. The survey highlighted phytotherapy plants used by the community, and greatly increased awareness by local livestock producers of poisonous plants for management purposes. We conclude that ethnobotanical knowledge, especially from the traditional community, is essential to understand livestock losses to toxic plants, and should be valued not only for reducing livestock losses, but also for cultural importance to the Kalunga communities in the Cerrado.

Pathologic and Immunohistochemical Evidence of Possible Francisellaceae among Aborted Ovine Fetuses, Uruguay.

The only genus of the Francisellaceae family known to contain species pathogenic to mammals is Francisella, for which reported cases in the Southern Hemisphere have been limited to Australia. We describe severe necrotizing and inflammatory lesions and intralesional immunohistochemical identification of Francisella sp. lipopolysaccharide among aborted ovine fetuses in Uruguay.

Epidemiological aspects of cattle tick fever in Brazil / Aspectos epidemiológicos da tristeza parasitária bovina no Brasil

The present work reviews the epidemiologic situation of Anaplasma marginale and Babesia spp. infections and the occurrence of cattle tick fever outbreaks in Brazil. In areas of tick fever enzootic instability, environmental conditions interfere with the development of Rhipicephalus (Boophilus) microplus: chilly winter in the southern region, floods in the Pantanal, and low humidity in the Caatinga. In contrast, the climatic conditions of stable zones (Cerrado, Amazon and Atlantic Forest biomes) favor tick development. In enzootic areas, tick fever is uncommon because the animals are in frequent contact with the parasite, acquiring immunity naturally during the period of innate resistance; however, outbreaks may occur when calves become infested by considerable numbers of infected ticks during this period or in adults raised in tick-free environments that become infested for the first time when transporting to stable areas. It is necessary to better understand the disease's risk factors under stable conditions and the implications of the mechanical and other vector transmission of A. marginale. To prevent tick fever outbreaks in Brazil, it is important to develop and use anaplasmosis and babesiosis vaccines in cattle from enzootic unstable regions, especially when animals are moved to stable areas.(AU)

O presente trabalho revisa a situação epidemiológica das infecções por Anaplasma marginale e Babesia spp. e a ocorrência de surtos de Tristeza parasitária bovina (TPB) no Brasil. Em áreas de instabilidade enzoótica, as condições ambientais interferem no desenvolvimento do Rhipicephalus (Boophilus) microplus: o frio do inverno na região Sul; as inundações no Pantanal; e a baixa umidade na Caatinga. Por outra parte, as condições climáticas das zonas de estabilidade (os biomas Cerrado, Amazônia e Mata Atlântica) favorecem o desenvolvimento do carrapato. A TPB não é comum nas áreas estáveis, porque os animais estão em contato frequente com os parasitas, adquirindo imunidade naturalmente. Podem, porém, ocorrer surtos quando um número considerável de carrapatos infectados infesta bezerros, durante o período de resistência inata, ou quando adultos que foram criados em ambientes livres de carrapatos infestam-se, pela primeira vez, ao serem transportados para áreas estáveis. É necessário entender melhor os fatores de risco da doença em condições de estabilidade e a implicação da transmissão de A. marginale de forma mecânica, ou por meio de vetores diferentes do carrapato. Para prevenir surtos de TPB, no Brasil, é necessário desenvolver e usar vacinas em bovinos de regiões de instabilidade, especialmente antes de transportá-los para áreas estáveis.(AU)

Cerebral infarction and thrombotic meningoencephalitis in a calf due to candidiasis / Infarto cerebral e meningoencefalite trombótica em um bezerro devido a candidíase

We described a case of cerebral infarction and thrombotic meningoencephalitis due to candidiasis in a seven-month-old calf. The death occurred three days after the onset of apathy, fever, and the head's lateral deviation to the left. Macroscopic changes in the brain consisted of asymmetry of telencephalic hemispheres; the right hemisphere was enlarged, causing cerebellar herniation. A focally extensive red area was observed on the surface of the right occipital lobe. At cross-sections of the fixed brain, the lesions revealed to be extensive, red-brown, soft or cavitated areas affecting the white and grey matter from the level of the thalamus to the cerebellum and compressing subjacent structures. Histologically, there was acute, coalescent, multifocal necrosupurative meningoencephalitis, associated with vasculitis, congestion, thrombosis, edema, infarction, and intralesional fungal hyphae. The diagnosis of cerebral infarction and thrombotic meningoencephalitis due to candidiasis was made by the pathological changes, the staining and morphological characteristics of the agent, and immunohistochemistry. The cerebral infarction and thrombotic meningoencephalitis in cattle can result from vascular lesions due to infection by Candida sp.; although uncommon, this case demonstrated that candidiasis should be part of a list of differential diagnoses of severe brain injuries in cattle.

Descreve-se um caso de infarto cerebral e meningoencefalite trombótica devido a candidíase em um bezerro de sete meses de idade. A morte ocorreu três dias após o início de apatia, febre e desvio lateral da cabeça para a esquerda. As alterações macroscópicas no cérebro consistiam em assimetria dos hemisférios telencefálicos; o hemisfério direito estava aumentado, causando herniação cerebelar. Uma extensa área vermelha focal foi observada na superfície do lobo occipital direito. Nos cortes transversais do encéfalo fixado, as lesões revelaram áreas extensas, marrom-avermelhadas, moles ou cavitadas, afetando a substância branca e cinzenta desde o nível do tálamo até o cerebelo e comprimindo as estruturas subjacentes. Histologicamente, havia meningoencefalite necrossupurativa multifocal aguda, coalescente, associada a vasculite, congestão, trombose, edema, infarto e hifas fúngicas intralesionais. O diagnóstico de infarto cerebral e meningoencefalite trombótica devido a candidíase foi feito pelas alterações patológicas, coloração e características morfológicas do agente e imuno-histoquímica. O infarto cerebral e meningoencefalite trombótica em bovinos pode resultar de lesões vasculares devido à infecção por Candida sp.; embora incomum, este caso demonstra que a candidíase deve fazer parte de uma lista de diagnósticos diferenciais de lesões cerebrais graves em bovinos.

Molecular of Anaplasma marginale Theiler (Rickettsiales: Anaplasmataceae) in horseflies (Diptera: Tabanidae) in Uruguay.

Anaplasma marginale is transmitted biologically by infected ticks or mechanically by biting flies and contaminated fomites. In tick-free areas, such as southern Uruguay, horseflies could be the principal vectors of this pathogen for bovines, causing anaplasmosis. The objective of this work was to detect the presence of A. marginale by MSP-5 PCR and Sanger sequencing in the most prevalent species of horseflies obtained using different collection methods in Colonia, Tacuarembó and Paysandú, Uruguay. Eight horsefly species were tested (Dasybasis missionum, Poeciloderas lindneri, Tabanus campestris, T. claripennis, T. fuscofasciatus, T. platensis, T. tacuaremboensis and T. triangulum); four species were found to be positive for A. marginale, with D. missionum and P. lindneri having the most frequent infections, while only one individual each of T. fuscofasciatus and T. tacuaremboensis was positive. Both D. missionum and P. lindneri were positive for A. marginale in tick-free areas, and the implications are discussed in this report.

Primary Photosensitization by Chamaecrista serpens in Santa Inês Sheep.

This study aimed to clarify the type of photosensitization induced by C. serpens and to verify if the plant remains toxic after being collected and stored. Eight crossbred sheep, aged between 6 and 36 months, were divided into three groups (G1 to G3). Over 30 days, daily, G1 received an exclusive diet of C. serpens, and G2 and G3 received 10 g/kg/BW and 20 g/kg/BW, respectively. Two other sheep were used as controls (CG). Before administration, the plant had been harvested every 15 days. Liver biopsies and blood samples were taken from all sheep on day zero and weekly. All sheep that received the plant developed clinical signs of photosensitization, and no changes were observed in the serum activities of AST and GGT. On day 30, all sheep except Ov1 from G1 and Ov7 were euthanized and necropsied. All sheep that received the plant developed clinical signs. Macroscopic or histologic lesions were not observed in the liver. Ov 1 recovered 13 days after the end of ingestion. These results demonstrated that C. serpens causes primary photosensitization. It is advisable to avoid grazing on pastures invaded by the plant or to remove them from the pastures immediately after observing the first signs.

Cestrum axillare (Solanaceae) poisoning in ruminants.

Cestrum axillare poisoning causes significant economic losses in farms of ruminant production due to a fatal acute hepatic disease. The consumption of C. axillare occurs on farms or pastures with a scarcity of feed or with dry forage. Epidemiological, clinical, and pathological data of poisoning outbreaks by C. axillare from 1953 to 2021 in grazing ruminants in southeastern Brazil are reported. A total of 68 bovines, two buffaloes, and two goats exhibited clinical signs and resulted in death due to C. axillare consumption, with 79% of the cases occurring during the dry period. Clinical signs were apathy, anorexia, ruminal arrest, arched back, and constipation with hard stools, sometimes with blood or mucus. Cases with neurological signs due to hepatic encephalopathy showed excitement, aggressiveness, drooling, staggering, and muscle tremors. The pathological findings included hepatocellular necrosis in the liver and microcavitations in the brain's white matter (status spongiosus). The hepatotoxins, carboxyparquin and parquin, were detected in C. axillare leaf samples collected from paddocks grazed by cattle in three southeastern Brazilian municipalities where outbreaks of C. axillare poisoning occurred. This is the first report of parquin and carboxyparquin in C. axillare.

A Retrospective Study of Pythiosis in Domestic Animals in Northeastern Brazil.

Pythiosis is an endemic disease in northeastern Brazil and we now report the epidemiological, clinical and pathological findings in a retrospective study of naturally occurring cases in domestic animals. From January 1985 to December 2020, the Laboratory of Animal Pathology of the Federal University of Campina Grande examined 13,542 tissue samples from necropsies and biopsies. Among these samples, 306 were diagnosed as pythiosis: 195 cases in horses, 75 in sheep, 19 in dogs, six in mules, four in cattle, three in cats, two in goats, one in a donkey and one in an ostrich. Affected equids had lesions in the skin, mammary glands and nasal cavities. Affected sheep had cutaneous, nasal and digestive lesions while cattle and goats had cutaneous lesions. Carnivores developed lesions, mainly in the alimentary tract, of sufficient severity to cause death or result in euthanasia. The single affected bird had lesions in the alimentary tract and surgical excision resulted in remission. The disease had a long and life-threatening clinical course in most affected species but resolved spontaneously in cattle. Clinical signs were directly related to the location of the lesions, which were invariably characterized by chronic inflammation associated with intralesional hyphae. Veterinary clinicians and pathologists should be familiar with the clinicopathological features of pythiosis and the wide range of susceptible animal species.

Poisoning by Nierembergia veitchii: Effects on vascular smooth muscle cells in the pathogenesis of enzootic calcinosis.

Vascular mineralization is a hallmark of enzootic calcinosis. Histopathological, ultrastructural, and immunohistochemical investigations were performed on the external carotid arteries of seven sheep naturally poisoned by Nierembergia veitchii. Histologically, moderate to marked hyperplasia of the tunica intima was observed without mineralization. The tunica media exhibited mild to severe mineralization and osteochondroid metaplasia. Sheep with enzootic calcinosis showed arterial overexpression of osteopontin and tissue-nonspecific alkaline phosphatase and immunolabeling for osteonectin and osteocalcin in both intima and media layers of the tested arteries. The main ultrastructural finding in the tunica media was a marked phenotypic change of vascular smooth muscle cells from a contractile phenotype (VSMC-C) into a synthetic phenotype (VSMC-S). In the tunica media, VSMC-S produced matrix and extracellular vesicles, forming mineralizable granules associated with arterial mineralization. VSMC-S were also present in the tunica intima, but matrix and extracellular vesicles and mineralization were not observed. The absence of matrix and extracellular vesicles in the intimal hyperplasia, even in the presence of noncollagenous bone proteins, tissue-nonspecific alkaline phosphatase, and vitamin D receptors, reinforces the hypothesis that the presence of matrix and extracellular vesicles are crucial for the development of vascular mineralization in enzootic calcinosis. It is proposed that the two different VSMC-S phenotypes in calcinosis are due to the expression of at least two genetically different types of these cells induced by the action of 1,25(OH)2D3.