DNA Methylation Status of the Estrogen Receptor α Gene in Canine Mammary Tumors.

Estrogen receptor α (ERα) has an important role in mammary carcinogenesis, prognosis, and treatment. In human and canine mammary cancer, the most aggressive tumors show loss of ERα expression, which in human breast cancer has been attributed to methylation of the cytosine followed by guanine (CpG) island within the estrogen receptor α gene ( ESR1) promoter. This study aimed to investigate the role of ESR1 CpG island (CGI) methylation in ERα expression in canine mammary tumors. Twenty-one canine mammary samples were sorted into three groups: malignant tumor (n = 9), benign tumor (n = 8), and normal gland (n = 4). Immunohistochemical analysis and reverse-transcription quantitative real-time PCR were performed to assess ERα expression and ESR1 mRNA levels. The methylation status was determined using sodium-bisulfite-treated DNA sequencing. All normal mammary glands and benign tumors showed high ERα expression (score range, 5-8). Six of the nine malignant tumors did not show ERα expression (score 0), two had score 2, and one had score 4. Lower ERα ( P < .005) and ESR1 mRNA levels ( P < .005) were found in malignant mammary tumors than in the other two groups. Canine ESR1 has an intragenic and non-promoter-associated CGI, different from humans. No significant variation in methylation percentage was observed among the groups, suggesting that ESR1 is not regulated by DNA methylation, unlike that in humans. This difference should be considered in further research using ERα as a biomarker for mammary tumors in canine studies on ERα-targeting therapy.

Detection of Neospora spp. and Sarcocystis neurona in amniotic fluid and placentas from mares.

The closely related apicomplexa protozoa, Toxoplasma gondii, Neospora spp., and Sarcocystis neurona, have all been associated with neurological and reproductive diseases in horses. However, there is limited data regarding the presence of these three parasites in equine placental tissues and amniotic fluid. The aim of the present report was to investigate the presence of the DNA of T. gondii, Neospora spp. and S. neurona in placentas and amniotic fluid in mares. Anti-T. gondii, anti- S. neurona and anti- Neospora spp. antibody titers were evaluated in 31 mares in the final third of pregnancy by indirect fluorescent antibody test (IFAT). The presence of parasite DNA in placentas and amniotic fluid was evaluated by polymerase chain reaction (PCR), using two target loci (ITS1 and Nc5). No antibodies to were identified nor was any T. gondii DNA detected in any mare. Antibodies to Neospora spp. were found in 6 mares (19.35 %) and DNA from this protozoan was detected in four placentas (12.9 %) and in five amniotic fluid samples (16.6 %). Antibodies to Sarcocystis spp. were detected in nine mares and S. neurona DNA was found in only one placenta (3.23 %). Our results suggest that the transplacental route may be a potential source of Neospora caninum infection in mares. Further studies are needed to understand the role of transplacental transmission in the epidemiology of these protozoa.

'Candidatus Mycoplasma haemosphiggurus' a novel haemoplasma species in orange-spined hairy dwarf porcupines (Sphiggurus villosus) from Southern Brazil.

In Brazil, the orange-spined hairy dwarf porcupine (Sphiggurus villosus) is widely distributed in the Atlantic Rainforest biome being amongst the most frequently road-killed animal. Porcupines may also be commonly found on forest borders and occasionally, near urban areas where human and domestic dogs injuries caused by its spines may occur. Therefore, the aims of this study were (a) to screen porcupines for TBD pathogens and haemoplasmas and (b) to identify the tick species parasitizing these rodents in Paraná State, southern Brazil. Blood and/or spleen samples were collected from nine orange-spined hairy dwarf porcupines. A total of 275 ticks (34 males, 11 females, 7 nymphs and 223 larvae) were collected from eight porcupines: Amblyomma longirostre, A. parkeri and Amblyomma spp. larvae. Two out of nine (22%; 95% CI: 3%-60%) porcupines were PCR-positive for haemoplasmas. All animals tested negative for Theileria/Babesia spp. and Ehrlichia/Anaplasma spp. by PCR. Phylogenetic and network analysis of the 16S and 23S rRNA gene fragments confirmed that animals were infected by a potentially novel haemotropic Mycoplasma sp. The name 'Candidatus Mycoplasma haemosphiggurus' is proposed for this novel organism that should be further fully characterized.

Transplacental transmission of Neospora caninum to lambs in successive pregnancies of naturally infected sheep in Southern Brazil.

This study aims to report the detection of N. caninum DNA in a newborn lamb (1) with neurological signs and congenital neosporosis and in a stillborn lamb (2), both born from the same ewe in a herd of Southern Brazil. The lambs were born during different pregnancies of a Suffolk ewe seropositive to N. caninum and seronegative to T. gondii. Histopathological lesions were observed only in the central nervous system of the lambs. The newborn lamb (1) showed mild and focal gliosis in the frontal lobe. In the hippocampal region of the stillborn lamb (2), lymphoplasmacytic perivascular cuffs and N. caninum cysts were observed in the cytoplasm of neurons and confirmed by IHC. PCR was performed using brain samples to detect the protozoa N. caninum and Toxoplasma gondii. The infection with N. caninum was confirmed in the newborn lamb (1) by PCR and in the stillborn lamb (2) by histopathology, immunohistochemistry, and PCR tests.

Safety and antiulcer efficacy studies of Achillea millefolium L. after chronic treatment in Wistar rats.

Achillea millefolium L. (Asteraceae), popularly known as yarrow, has been used in folk medicine to treat complaints such as inflammation, pain, wounds, hemorrhages and gastrointestinal disturbances. The aim of the present study was to assess the safety and efficacy of the aqueous extract (AE) of the plant after chronic exposure. Indeed, the AE was effective in protecting the gastric mucosa against acute gastric lesions induced by ethanol and indomethacin and in healing chronic gastric lesions induced by acetic acid with (ED(50)=32 mg/kg, p.o.). Safety studies were performed in female and male Wistar rats treated daily with AE (0.3-1.2 g/kg, p.o./day) or vehicle (water, 10 ml/kg/day) for 28 or 90 consecutive days. Satellite groups consisted of animals sacrificed 30 days after the end of these treatments. Clinical observations, body and organ weight measurements, gross autopsy, hematology, clinical biochemical and histopathological examinations were performed. Slight changes in liver weight, cholesterol, HDL-cholesterol and glucose were observed in male and female animals. These changes were not correlated with dose or time of exposure of the animals to the AE. Overall, the results show the antiulcer potential of the aerial parts of the Achillea millefolium which is accompanied by no signs of relevant toxicity even at very long chronic exposure.

Oxidative stress in hoof laminar tissue of horses with lethal gastrointestinal diseases.

Tissue damage caused by oxidative stress is involved in the pathogenesis of several diseases in animals and man, and is believed to play a role in the development of laminitis in horses. The aim of this study was to investigate the oxidative stress associated with laminar lesions in horses with lethal gastrointestinal disorders. Laminar tissue samples of the hoof of 30 horses were used. Tissue samples were divided as follows: six healthy horses (control group-CG), and 24 horses that died after complications of gastrointestinal diseases (group suffering from gastrointestinal disorders-GDG). Superoxide dismutase (SOD2) and nitrotyrosine immunostaining and the severity of laminar lesions were evaluated. Presence of laminar lesions and immunostaining for nitrotyrosine and SOD2 were only evident in horses from the GDG group. Thus, oxidative stress may play a role in the pathogenesis of laminar lesions secondary to gastrointestinal disorders.

Systemic leukopenia, evaluation of laminar leukocyte infiltration and laminar lesions in horses with naturally occurring colic syndrome.

The present study was aimed at identifying laminar lesions and leukocyte infiltration in hoof laminar tissue of horses with colic syndrome and its correlation with the total leukocyte count before death. Six healthy horses were used as control group (CG), and eighteen horses with lethal gastrointestinal disease were divided into two groups: leukopenic group (LG) with seven leukopenic horses, and non-leukopenic group (NLG) with 11 horses with total leukocyte count within reference range for the species. Leukocyte infiltration was examined by immunohistochemistry. Laminar lesions were observed in both LG and NLG, with no differences in severity between them. LG showed increase of the leukocyte infiltration in the hoof laminar tissue, when compared to CG and NLG. Horses with severe colic syndrome (LG and NLG) developed intense laminar lesions without clinical signs of laminitis, with increased leukocyte infiltration. However, the LG demonstrated an even higher increase of leukocyte infiltration compared to both CG and NLG.

Ethyl 2-cyanoacrylate tissue adhesive in partial lobectomy in rats.

OBJECTIVE: To determine the efficacy of ethyl 2-cyanoacrylate adhesive in repairing the lung parenchyma after partial lobectomy in rats, in terms of hemostasis/aerostasis, scarring, and surgical time. METHODS: The study involved 30 Wistar rats, randomly divided into five groups (one control group and four study groups). In the study groups, the lung parenchyma was repaired with either cyanoacrylate adhesive or surgical suture following resection of a small or large fragment (25% or 50%, respectively) of the left caudal lung lobe. RESULTS: Surgical time and hemostasis time were shorter in the two groups treated with the adhesive than in the two submitted to suture. There were no significant differences among the groups regarding specific lung compliance. Adherences and inflammatory reactions were more severe in the groups submitted to suture. CONCLUSIONS: In this study, the use of cyanoacrylate adhesive helped reduce the surgical time and the intensity of inflammatory reactions, as well as preserving lung compliance. Cyanoacrylate adhesives should be considered an option for lung parenchyma repair, decreasing the risk of complications after partial lobectomy in humans.

Adesivo cirúrgico de etil-2-cianoacrilato em lobectomia parcial em ratos / Ethyl 2-cyanoacrylate tissue adhesive in partial lobectomy in rats

OBJETIVO: Determinar experimentalmente a eficácia do adesivo cirúrgico de etil-2-cianoacrilato na reparação do parênquima pulmonar após lobectomias parciais em ratos em relação a hemostasia/aerostasia, cicatrização e tempo cirúrgico. MÉTODOS: O estudo envolveu 30 ratos Wistar, divididos aleatoriamente em cinco grupos (grupo controle e quatro grupos de estudo. Nos grupos de estudo, o reparo do parênquima pulmonar foi realizado ou com o adesivo ou por sutura após lobectomia parcial de um fragmento pequeno ou grande (25 por cento ou 50 por cento, respectivamente) do lobo inferior caudal esquerdo. RESULTADOS: O tempo cirúrgico e o tempo de hemostasia foram menores nos grupos submetidos ao uso do adesivo. Não houve diferenças significativas na complacência pulmonar específica entre os grupos. Aderências e reações inflamatórias foram mais severas nos grupos submetidos a sutura. CONCLUSÕES: Neste estudo, o uso de adesivo de cianoacrilato ajudou a reduzir o tempo cirúrgico e a intensidade de reações inflamatórias, assim como preservou a complacência pulmonar. Adesivos de cianoacrilato devem ser considerados como uma opção no reparo do parênquima pulmonar, diminuindo o risco de complicações após lobectomia parcial em humanos.

OBJECTIVE: To determine the efficacy of ethyl 2-cyanoacrylate adhesive in repairing the lung parenchyma after partial lobectomy in rats, in terms of hemostasis/aerostasis, scarring, and surgical time. METHODS:The study involved 30 Wistar rats, randomly divided into five groups (one control group and four study groups). In the study groups, the lung parenchyma was repaired with either cyanoacrylate adhesive or surgical suture following resection of a small or large fragment (25 percent or 50 percent, respectively) of the left caudal lung lobe. RESULTS: Surgical time and hemostasis time were shorter in the two groups treated with the adhesive than in the two submitted to suture. There were no significant differences among the groups regarding specific lung compliance. Adherences and inflammatory reactions were more severe in the groups submitted to suture. CONCLUSIONS: In this study, the use of cyanoacrylate adhesive helped reduce the surgical time and the intensity of inflammatory reactions, as well as preserving lung compliance. Cyanoacrylate adhesives should be considered an option for lung parenchyma repair, decreasing the risk of complications after partial lobectomy in humans.

Intoxicaçäo experimental por Phalaris angusta (Gramineae) em bovinos / Experimental poisoning by Phalaris angusta (Gramineae) in cattle

Para investigar os efeitos da ingestäo de diferentes quantidades da planta Phalaris angusta em bovinos, oito bezerros, com idade variando entre 6-8 meses, foram divididos em 4 grupos com 2 animais cada. Os animais do grupo I receberam somente P. angusta na alimentaçäo, enquanto que os animais do grupo II receberam P. angusta (75 por cento), aveia (Avena sativa) e azevém (Lolium multiflorum) (25 por cento). Os animais do grupo III receberam P. angusta (50 por cento), aveia e azevém (50 por cento) e os animais do grupo IV receberam somente aveia e azevém e serviram como controles. Todos os animais que ingeriram P. angusta adoeceram. Um animal do grupo I morreu 34 dias após o início da ingestäo da planta e os outros animais foram sacrificados, in extremis, em um período que variou de 18 a 32 dias após o início do experimento. Os principais sinais clínicos observados foram alterações de locomoçäo, tremores generalizados, quedas e crises convulsivas. Alterações macroscópicas foram observadas apenas no encéfalo e eram caracterizadas por focos de coloraçäo verde-azulada no tálamo, mesencéfalo e medula oblonga. Microscopicamente observou-se pigmento granular marrom-amarelado no citoplasma de neurônios das regiões macroscopicamente afetadas. Alterações ultra-estruturais consistiram de lisossomos contendo material com densidade e orientaçäo variáveis. A quantidade de P. angusta ingerida näo foi um fator determinante na gravidade do quadro clínico, nem na intensidade das lesões observadas. A intensidade dos sinais clínicos também näo teve uma relaçäo direta com a severidade das lesões macro e microscópicas. Phalaris angusta demonstrou ter açäo exclusivamente neurotóxica e deve ser considerada no diagnóstico diferencial em casos de animais com sinais clínicos de origem nervosa, consistentes com síndrome tremorgênica