Nanocomposite Based on Bacterial Cellulose and Silver Nanoparticles Improve Wound Healing Without Exhibiting Toxic Effect.

Wound healing is an important and complex process, containing a multifaceted process governed by sequential yet overlapping phases. Certain treatments can optimize local physiological conditions and improve wound healing. Silver nanoparticles (AgNP) are widely known for their antimicrobial activity. On the other hand, bacterial cellulose (BC) films have been used as a dressing that temporarily substitutes the skin, offering many advantages in optimizing wound healing, in addition to being highly biocompatible. Considering the promising activities of AgNP and BC films, the present study aimed to evaluate the wound healing activity in Wistar Hannover rats using a nanocomposite based on bacterial cellulose containing AgNP (AgBC). In a period of 21 days, its influence on the wound area, microbial growth, histopathological parameters, and collagen content were analyzed. In addition, toxicity indicators were assessed, such as weight gain, water consumption, and creatinine and alanine transaminase levels. After 14 days of injury, the animals treated with AgBC showed a significant increase in wound contraction. The treatment with AgBC significantly reduced the number of microbial colonies compared to other treatments in the first 48 h after the injury. At the end of the 21 experimental days, an average wound contraction rate greater than 97 % in relation to the initial area was observed, in addition to a significant increase in the amount of collagen fibers at the edge of the wounds, lower scores of necrosis, angiogenesis and inflammation, associated with no systemic toxicity. Therefore, it is concluded that the combination of preexisting products to form a new nanocomposite based on BC and AgNP amplified the biological activity of these products, increasing the effectiveness of wound healing and minimizing possible toxic effects of silver.

Evaluation of the antiseptic and wound healing potential of polyhexamethylene guanidine hydrochloride as well as its toxic effects.

The synthetic polyhexamethylene guanidine hydrochloride (PHMGH) polymer presents antifungal and antimicrobial activities in vitro. However, in vivo reports regarding its antiseptic and healing activity are scarce in the scientific literature. Thus, the present study aimed to evaluate the antimicrobial and healing effects, as well as toxicological parameters, of a topical solution containing 0.5% PHMGH (Akwaton®) in the treatment of superficial skin wounds experimentally induced on the dorsum of rodents. In addition, non-clinical safety studies were also conducted for use in human health, such as acute oral toxicity and genotoxicity tests. Animals did clinically not present dermatitis. After two days of topical treatment, PHMGH showed a significant antiseptic effect compared to the untreated group, reducing the number of colony-forming units by 72%, reaching 100% on the fourth day of treatment. The animals treated with PHMGH showed a significant area reduction of the skin lesions in relation to the untreated group, indicating a healing effect of the polymer. Moreover, PHMGH treatment led to a significant increase in fibroblasts when compared to the untreated group, revealing its healing action. No significant differences were observed between the biochemical indicators of hepatoxicity and nephrotoxicity, nor genotoxicity between the PHMGH-treated and the negative control groups. The results of acute oral toxicity showed that PHMGH at 5% presents a lethal dose 50% greater than the 2000 mg/kg. At a concentration of 5%, PHMGH did not show genotoxicity nor cytotoxicity at doses up to 1500 mg/kg through the micronucleus assay in mice. Therefore, 0.5% PHMGH showed an antimicrobial and healing effect, with no toxicity, and could be a promising adjunct in the microbial control of healing wounds.

Ovarian dysgerminoma causing multifocal metastases in young bitch / Disgerminoma ovariano causando metástases multifocais em cadela jovem

The ovarian neoplasias are unusual in canine species, mainly originated from germinative cells. Among these, dysgerminoma is a rare neoplasm in bitches, predominantly affecting senile and small and medium breeds. They are large, rounded, smooth surface, painless and firm in consistency. The clinical signs commonly demonstrated by patients with this neoplasia are abdominal distension and secondary signs of hyperestrogenism. The diagnosis should be based on the association of history, clinical signs, radiographic, ultrasonographic, cytological, histopathological and immunohistochemical exams. The treatment is the surgical castration and metastases are uncommon. Due to the rarity in bitches, specifically the young animals, this study aimed to describe a case of left ovarian dysgerminoma, of considerable size and adherence in adjacent organs, in a Brazilian Fila, whose showed considerable increase in abdominal volume and progressive weight loss. The diagnosis was confirmed by means of complementary x-ray, ultrasound, histopathological and immunohistochemical tests. Even with the recommended surgical treatment (castration), the patient demonstrated severe respiratory symptoms and increased abdominal volume after two months, and the complementary exams suggested pulmonary and multifocal metastases and the tutor chose euthanasia. As presented by the results, although rare in bitches, dysgerminoma should be included in the differential of ovarian neoplasias, including the young ones.

As neoplasias ovarianas são de ocorrência incomum na espécie canina, principalmente as originadas de células germinativas. Dentre estas, o disgerminoma é uma neoplasia rara em cadelas, acometendo predominantemente fêmeas senis e de raças pequenas e médias. Apresentam-se como massas grandes, arredondadas, de superfície lisa, indolores e de consistência firme. Os sinais clínicos comumente demonstrados por pacientes com este tipo neoplásico incluem distensão abdominal e sinais secundários de hiperestrogenismo. O diagnóstico deve ser baseado na associação do histórico, sinais clínicos, exames radiográficos, ultrassonográficos, citológicos, histopatológicos e imunohistoquímicos. O tratamento de escolha é a castração cirúrgica, sendo que as metástases são incomuns. Perante a raridade em cadelas, em específico as jovens, este relato teve como propósito discorrer um caso de disgerminoma ovariano esquerdo, de tamanho considerável e aderência em órgãos adjacentes, em uma Fila Brasileira, cuja sintomatologia incluiu considerável aumento de volume abdominal e emagrecimento progressivo. O diagnóstico foi confirmado por meio de exames complementares de raios-x, ultrassom, histopatológico e imunohistoquímicos. Mesmo com a instituição dotratamento cirúrgico preconizado (castração), a paciente demonstrou grave sintomatologia respiratória e aumento de volume abdominal após dois meses e, como os exames complementares foram sugestivos de metástases pulmonares e multifocais, o tutor optou pela eutanásia. Diante dos resultados, pode-se admitir que apesar de raro em cadelas, o disgerminoma deve ser incluído no diferencial das neoplasias ovarianas, incluindo as jovens.

Effect of systemic administration of phytosterol on lacrimal quality of dogs / Efeito da administração sistêmica de fitoesterol na qualidade lacrimal de cães

The tear lipid layer (oily outer layer) reduces evaporation and prevents tear overflow. In dogs, reductions in the lipid components of this layer (cholesterol, triglycerides and phospholipids) can cause eye serious diseases. In this way, the tear crystallization test analyzes the lacrimal quality, however, it is less used in veterinary. As phytosterol reduces blood cholesterol, the objective of this study was to investigate, through the tear crystallization test, whether the systemic administration of this drug influences the lacrimal quality of healthy dogs and, in addition, to verify differences in the interpretation of the ophthalmic test between different evaluators. Eight beagles, healthy, of both sexes, young and adults, without clinical ophthalmic signs apparent were selected. Basal lacrimal samples (D0) were collected from the right and left eye of all animals with glass capillary tube and arranged on a glass slide for scanning the images and subsequent microscopic analysis. Subsequently, all were medicated with the phytosterol (Collestra® 650 mg: 1 capsule, orally, every 12 hours, for 15 days). After seven (D7) and fifteen (D15) days of this systemic administration, the tear crystallization test in both eyes of all dogs was again performed for statistical comparison with the baseline results. The photographs of the slides were classified by four evaluators (AV1 and AV2 with professional experience in ophthalmology and AV3 and AV4 without previous professional experience in ophthalmology), following standards established by Rolando (1984). The results were statistically verified by analysis of simple variance (ANOVA One-Way). There was no statistical difference in the tear crystallization test between the established periods and in relation to the different ophthalmic test evaluators (p≤0.05). Although phytosterols reduce blood cholesterol levels, it was observed in the present study that these drugs when administered systemically did not interfere in the tear lipid layer and, consequently, in the lacrimal quality of healthy dogs, and may be prescribed as lipid-lowering agents for patients with ocular diseases, especially the lacrimal ones.

A camada lipídica lacrimal (camada externa oleosa) reduz a evaporação e previne o transbordamento lacrimal. Em cães, reduções nos componentes lipídicos desta camada (colesterol, triglicérides e fosfolipídios) podem causar doenças graves nos olhos. Desta forma, o teste de cristalização lacrimal analisa a qualidade lacrimal, no entanto, é menos utilizado em veterinária. Como o fitoesterol reduz o colesterol sanguíneo, o objetivo deste estudo foi investigar, através do teste de cristalização lacrimal, se a administração sistêmica deste fármaco influencia na qualidade lacrimal de cães hígidos e, além disso, verificar diferenças na interpretação do teste oftalmológico entre diferentes avaliadores. Oito beagles, saudáveis, de ambos os sexos, jovens e adultos, sem sinais oftalmológicos clínicos aparentes foram selecionados. Amostras lacrimais basais (D0) foram coletadas do olho direito e esquerdo de todos os animais, com tubo capilar de vidro e, dispostas em lâmina de vidro para escaneamento das imagens e posterior análise microscópica. Ato contínuo todos foram medicados com o fitoesterol (Collestra® 650 mg: 1 cápsula, por via oral, a cada 12 horas, durante 15 dias). Após sete (D7) e quinze (D15) dias desta administração sistêmica, o teste de cristalização lacrimal em ambos os olhos de todos os cães foi novamente realizado para comparação estatística com os resultados basais. As fotografias das lâminas foram classificadas por quatro avaliadores (AV1 e AV2 com experiência profissional em oftalmologia e AV3 e AV4 sem experiência profissional prévia em oftalmologia), seguindo padrões estabelecidos por Rolando (1984). Os resultados foram estatisticamente verificados pela análise de variância simples (ANOVA One-Way). Não houve diferença estatística no teste de cristalização lacrimal entre os períodos estabelecidos e em relação aos diferentes avaliadores de teste oftalmológico (p≤0,05). Embora os fitoesteróis reduzam os níveis de colesterol no sangue, observou-se no presente estudo que esses fármacos quando administrados sistemicamente não interferiram na camada lipídica da lágrima e, consequentemente, na qualidade lacrimal de cães hígidos, podendo ser prescritos como agentes hipolipemiantes para pacientes com doenças oculares, especialmente as lacrimais.

Video-assisted ovariohysterectomy in domestic cats (Felis catus, Linnaeus, 1758) using two access portals.

PURPOSE: To describe video-assisted ovariohysterectomy (OHE) with two portals access in adult intact queens. METHODS Fifty-two females cats were used. A 4 mm cannula was positioned in the abdomen through an incision close to the umbilicus (first portal), and a pneumoperitoneum was established. A second portal was positioned in the midline of the pre-pubic region. Females were positioned in right lateral recumbency to locate the left ovarian pedicle, and the uterine horn was held by a transcutaneous suture. The pedicle was cauterized and incised. The procedure was then performed on the contralateral ovary. The ovaries were exteriorized from the abdomen, along with the uterus, through the second access point. The uterine body was exposed, fixed and sectioned, and the abdominal incisions were sutured. RESULTS Surgeries were performed in an average of 41.4±14.2 minutes. The main complications included hypotension (7.7%) and subcutaneous emphysema (7.7%), and 13.5% of the surgeries were converted to laparotomy. CONCLUSION Ovariohysterectomy using a video-assisted technique and two access portals is safe, has minimal risks and is effective for the spaying of queens.

Video-assisted ovariohysterectomy in domestic cats (Felis catus, Linnaeus, 1758) using two access portals

PURPOSE : To describe video-assisted ovariohysterectomy (OHE) with two portals access in adult intact queens. METHODS Fifty-two females cats were used. A 4 mm cannula was positioned in the abdomen through an incision close to the umbilicus (first portal), and a pneumoperitoneum was established. A second portal was positioned in the midline of the pre-pubic region. Females were positioned in right lateral recumbency to locate the left ovarian pedicle, and the uterine horn was held by a transcutaneous suture. The pedicle was cauterized and incised. The procedure was then performed on the contralateral ovary. The ovaries were exteriorized from the abdomen, along with the uterus, through the second access point. The uterine body was exposed, fixed and sectioned, and the abdominal incisions were sutured. RESULTS Surgeries were performed in an average of 41.4±14.2 minutes. The main complications included hypotension (7.7%) and subcutaneous emphysema (7.7%), and 13.5% of the surgeries were converted to laparotomy. CONCLUSION Ovariohysterectomy using a video-assisted technique and two access portals is safe, has minimal risks and is effective for the spaying of queens.

Description of six autochthonous cases of canine visceral leishmaniasis diagnosed in Pedregulho (São Paulo, Brazil).

Visceral leishmaniasis is an infectious disease of chronic, emerging and zoonotic nature that presents various degrees of severity. In Brazil, this illness is caused by Leishmania infantum (Leishmania chagasi), which is transmitted by the bite of the sand fly Lutzomyia longipalpis, and dogs are its main reservoir. Given the increasing spread of this disease across Brazil, the aim of this study was to report on six cases of canine visceral leishmaniasis, diagnosed in June 2013, in the city of Pedregulho, State of São Paulo, considered to be a non-endemic area and free of phlebotomine sand flies. The diagnosis was based on clinical signs of the patients and additional tests (serological and parasitological). It was concluded that the diagnosis of leishmaniasis is complex because the clinical signs are similar to other systemic diseases, thus justifying the importance of parasitological test of bone marrow, considered "gold standard", in the confirmation of the disease. In addition, the area was not, until now, considered risk place, despite notification.

Description of six autochthonous cases of canine visceral leishmaniasis diagnosed in Pedregulho (São Paulo, Brazil) / Descrição de seis casos autóctones de leishmaniose visceral canina, diagnosticados em Pedregulho (São Paulo, Brasil)

Visceral leishmaniasis is an infectious disease of chronic, emerging and zoonotic nature that presents various degrees of severity. In Brazil, this illness is caused by Leishmania infantum (Leishmania chagasi), which is transmitted by the bite of the sand fly Lutzomyia longipalpis, and dogs are its main reservoir. Given the increasing spread of this disease across Brazil, the aim of this study was to report on six cases of canine visceral leishmaniasis, diagnosed in June 2013, in the city of Pedregulho, State of São Paulo, considered to be a non-endemic area and free of phlebotomine sand flies. The diagnosis was based on clinical signs of the patients and additional tests (serological and parasitological). It was concluded that the diagnosis of leishmaniasis is complex because the clinical signs are similar to other systemic diseases, thus justifying the importance of parasitological test of bone marrow, considered "gold standard", in the confirmation of the disease. In addition, the area was not, until now, considered risk place, despite notification.

A leishmaniose visceral é uma doença infecciosa de caráter crônico, emergente e zoonótico, apresentando variados graus de severidade. No Brasil, tal enfermidade é causada pela Leishmania infantum (Leishmania chagasi), a qual é transmitida pela picada de flebótomo Lutzomyia longipalpis, tendo o cão como o principal reservatório. Perante a crescente distribuição da doença no território brasileiro, o objetivo do presente trabalho é relatar seis casos de leishmaniose visceral canina, diagnosticados em Junho de 2013, no município de Pedregulho, Estado de São Paulo, considerado área não endêmica e livre de flebotomíneos. O diagnóstico foi baseado nos sinais clínicos dos pacientes e exames complementares (sorológicos e parasitológicos). Concluiu-se que o diagnóstico da leishmaniose é complexo pelo fato dos sinais clínicos serem semelhantes a outras enfermidades sistêmicas, justificando assim a importância do teste parasitológico de medula óssea, considerado padrão ouro na confirmação da doença. Além disso, a área não foi, até o presente momento, considerada local de risco, apesar da notificação.

Endodontics pastes formulated with copaiba oil: action on oral microbiota and dentin bridge formation in dogs / Pastas endodônticas formuladas com óleo de copaíba: ação na microbiota bucal e formação de ponte dentinária em cães

This study analysed the effect of pastes formulated with calcium hydroxide P.A. and different vehicles (saline solution - paste A and Copaifera langsdorffii Desfon oil - paste B) on oral microorganisms and dentin bridge formation in dogs. The antimicrobial action of the pastes and their components was analysed by the minimum inhibitory concentration in agar gel technique. The components were diluted and tested on fifteen standard strains of microorganisms associated with endodontic diseases. The microorganisms were cultivated and after incubation data was analysed using One-Way ANOVA and Turkey's test (P≤0.05). Four superior incisors of ten animals were used to evaluate dentin bridge formation. Two incisors were capped with paste A (GA) and two with paste B (GB). After 90 days, the teeth were extracted for histological analysis and the degree of dentin bridge formation evaluated. Data was analysed by the Kruskal-Wallis test (P<0.05). The pastes and their components were classified in the following decreasing order of antimicrobial action: calcium hydroxide P.A., paste A, paste B and Copaifera langsdorffii Desfon oil. Calcium hydroxide P.A. showed significantly higher antimicrobial action than the pastes or their vehicles. No significant difference was observed between the two pastes in dentin bridge formation. Based on the microorganisms studied, it can be concluded that the pastes analysed showed similar antimicrobial potential but differed significantly from their individual components. No significant difference was observed in dentin bridge formation between the different pastes tested.

Foi avaliada a ação de pastas formuladas com hidróxido de cálcio P.A. e diferentes veículos (solução fisiológica - pasta A e óleo de Copaifera langsdorffii Desfon - pasta B) sobre microrganismos bucais e formação de ponte dentinária em cães. A ação antimicrobiana das pastas e de seus componentes individuais foi avaliada pela técnica da concentração inibitória mínima pela diluição em ágar. Os materiais foram diluídos e avaliados contra quinze cepas padrão de microrganismos relacionados a doenças endodônticas. Os microrganismos foram cultivados e, após a incubação, os dados foram analisados (Anova One-Way, Tukey, P≤0,05). Para a avaliação da formação de ponte dentinária, quatro incisivos superiores de dez animais foram tratados, sendo dois capeados com pasta A (GA) e dois com a pasta B (GB). Após 90 dias, os dentes foram extraídos para obtenção de cortes histológicos, com o objetivo de se avaliar o grau de formação de ponte dentinária. Os dados obtidos foram submetidos à análise estatística (Kruskal-Wallis, P<0,05). Segundo a ação antimicrobiana, os materiais foram classificados em ordem decrescente: hidróxido de cálcio P.A., pasta A, pasta B e óleo de Copaifera langsdorffii Desfon, com diferenças estatisticamente significantes entre o hidróxido de cálcio P.A., as pastas e os veículos. Ao final do estudo, observou-se que as duas pastas avaliadas foram semelhantes quanto à formação de ponte dentinária. Considerando-se os microrganismos selecionados, conclui-se que as pastas avaliadas foram semelhantes entre si em termos de potencial antimicrobiano, diferindo dos componentes individuais. Em relação à formação de ponte dentinária, não foram observadas diferenças significantes entre os dentes tratados com as diferentes pastas.