Análise morfométrica comparada entre Anuros endêmicos do Brasil e a espécie invasora Lithobates catesbeianus

Anfíbios são indicadores ambientais potencialmente confiáveis e eficientes. Estudos referentes a morfologia de leucócitos de anuros são limitados, com poucos estudos morfometricos disponíveis em literatura. O presente estudo empregou técnicas morfometricas para caracterizar leucócitos de anuros Neotropicais brasileiros selecionados e compara-los com a espécie exótica rã-touro (Lithobates catesbeianus), família Ranidae. Esfregaços sanguíneos de 28 espécimes pertencentes a seis gêneros diferentes (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus e Proceratophrys) foram comparados com amostras de esfregacos de L. catesbeianus. A média do diâmetro dos leucócitos foi calculada por um software de análise de imagens. One-way e teste de Bonferroni foram utilizados para avaliação estatística. Linfócitos, neutrófilos, eosinófilos e basófilos mostraram-se significativamente menores que os valores de referência reportados em outros gêneros de anfíbios, incluindo Lithobathes; por outro lado, a média do diâmetro dos monócitos não demonstrou variação significativa entre os gêneros. Esse e o primeiro estudo de avaliação morfometrica de leucócitos em espécies de anuros brasileiros. Nossos resultados sugerem que a separação geográfica possivelmente influencia a morfometria leucocitaria.Amphibians are potentially reliable and efficient bioindicators. Existing anuran white blood cell morphology studies are limited, with only a few morphometric studies available. We employed morphometric techniques to characterize leukocytes of selected Neotropical anurans from Brazil and compare our findings with the exotic American Bullfrog (Lithobates catesbeianus), genus Ranidae. We compared blood smears of 28 specimens from six different genera (Hyla, Phyllomedusa, Hypsiboas, Scinax, Physalaemus, and Proceratophrys) with samples from L. catesbeianus. Leukocyte average diameter was calculated by an image analysis software. One-way analyses of variance and Bonferroni tests were used on statistical analyses. Lymphocytes, neutrophils, eosinophils, and basophils were significantly smaller than the reference ranges reported for other amphibian genera, including Lithobathes, whereas monocyte diameters did not differ significantly between genera. This is the first study to evaluate leukocyte morphometrics of Brazilian anuran species. Our findings suggest that geographical separation could possibly influence leukocyte morphometry

Tumoral Resection followed by Blepharoplasty in Squamous Cell Carcinoma Treatment in the Lower Eyelid of a Horse

Background: Equines are routinely subjected to enucleation due to palpebral tumors. Blepharoplasties in horses, especially in the lower eyelid, are rarely performed due to the difficulty of sliding once the tissue around the eyes presents low mobility. Defects involving more than 50% of the lower eyelid is considered challenging after tumor removal. Squamous cell carcinoma (SCC) is the second most reported neoplasm in horses, being very common in regions of the lower eyelid, third eyelid, sclera and or cornea. The aim of this study is to present the Destro VY skin advancement flap as a blepharoplasty technique performed after surgical excision of a SCC, with total commitment of the lower eyelid, completely covering the right eye of a mare.Case: A 8-year-oldmarePaint Horse, weighing420 kg,was referred for evaluation of tumor tissue of 6.0 x 4.0 x 2.0 cm, with nodular and ulcerated appearance, involving the right lower eyelid, in its total extension and completely covering the eye, without adhering to it. Considering the initial suspicion of SCC, the treatment strategy performed was surgical eyelid excision and maintenance of the eye, followed by blepharoplasty as an attempt to reconstruct the eyelid. Under general inhalation anesthesia, the animal was placed in left lateral recumbency, when the surgical region was prepared and local anesthetic block was performed. After antisepsis, a skin incision was made circumscribing the tumor, respecting a margin of 10 mm apart and excision of all visible tumor tissue was performed followed by intralesional ozone therapy. Blepharoplasty was performed to cover the portions of the exposed lacrimal and zygomatic bones, as well as correction of the eyelid aesthetics. For this, Destro VY skin advancement flap was performed for reconstruction of the lower eyelid. An incision of approximately 7 cm in V-shaped skin was performed, and the subcutaneous tissue under the V was dissected, maintaining a central pedicle, responsible for the vascularization of the flap, which was slid, approximately 20 mm, towards the eye. After obtaining the desired skin approximation, Y-suture was performed, covering the exposed bone and reconstructing the lower eyelid. In the postoperative period, local instillation of mitomycin eye drops and systemic meloxicam administration were instituted. The mare had her vision restored, presenting satisfactory morpho functional and aesthetic results and no tumor recurrence during 1-year of follow-up. Discussion: The repair of lower eyelid imperfections is challenging, especially when they have large defects, and there are no reports of performing the Destro VY skin advancement flap technique in horses for lower eyelid reconstruction. In this case, the importance of the blepharoplasty technique is emphasized, avoiding enucleation, preserving horse’s vision and aesthetics. In addition, aiming to avoid tumor recurrence, especially if surgical safety margins can not be achieved, other complementary treatments should be associated, including intralesional ozone therapy, mitomycin, an antineoplastic chemotherapy drug, and meloxicam, a COX-2 selective, non-steroidal anti-inflammatory drug, as performed in this study. It is concluded that the use of the Destro VY skin advancement flap technique for reconstruction of the external lamella in cases of SCC in the lower eyelid of horses is a feasible technique, which preserves the animal's vision, as well as aesthetics. The safety margin in the surgical excision of the SCC and the association of complementary therapies in the resolution of the condition are important points also to be considered.Keywords: epidermoid carcinoma, equine, ophthalmology, plastic surgery, skin tumor. Título: Exérese tumoral seguida de blefaroplastia no tratamento de carcinoma de células escamosas em pálpebra inferior de equino

Oviductal transcriptional profiling of a bovine fertility model by next-generation sequencing

In cattle, the oviduct plays a fundamental role in the reproductive process. Oviductal functions are controlled by the ovarian sex steroids: estradiol and progesterone. Here, we tested the hypothesis that the exposure to contrasting sex steroid milieus differentially impacts the oviductal transcriptional profile. We manipulated growth of the pre-ovulatory follicle to obtain cows that ovulated a larger (LF group) or a smaller (SF group) follicle. The LF group presented greater proestrus/estrus concentrations of estradiol and metaestrus concentrations of progesterone (Gonella-Diaza et al. 2015 [1], Mesquita et al. 2014 [2]). Also, the LF group was associated with greater fertility in timed-artificial insemination programs (Pugliesi et al. 2016 [3]). Cows were slaughtered on day 4 of the estrous cycle and total RNA was extracted from ampulla and isthmus fragments and analyzed by RNAseq. The resulting reads were mapped to the bovine genome (Bos taurus UMD 3.1, NCBI). The differential expression analyses revealed that 325 and 367 genes in ampulla and 274 and 316 genes in the isthmus were up-regulated and down-regulated in LF samples, respectively. To validate the RNAseq results, transcript abundance of 23 genes was assessed by qPCR and expression patterns were consistent between the two techniques. A functional enrichment analysis was performed using Database for Annotation, Visualization and Integrated Discovery (DAVID) software. Processes enriched in the LF group included tissue morphology changes (extracellular matrix remodeling), cellular changes (proliferation), and secretion changes (growth factors, ions and metal transporters). An overview of the gene expression data was deposited in the NCBI's Gene Expression Omnibus (GEO) and is accessible through the accession number GSE65681. In conclusion, differences in the peri-ovulatory sex steroid milieu modify the oviductal gene expression profiles. Such differences may be associated with the greater fertility of the LF cows. This dataset is useful for further investigations of the oviductal biology and the impact of sex-steroid on the female reproductive tract

Size of the Ovulatory Follicle Dictates Spatial Differences in the Oviductal Transcriptome in Cattle

In cattle, molecular control of oviduct receptivity to the embryo is poorly understood. Here, we used a bovine model for receptivity based on size of the pre-ovulatory follicle to compare oviductal global and candidate gene transcript abundance on day 4 of the estrous cycle. Growth of the pre-ovulatory follicle (POF) of Nelore (Bos indicus) cows was manipulated to produce two groups: large POF large corpus luteum (CL) group (LF-LCL; greater receptivity) and small POF-small CL group (SF-SCL). Oviductal samples were collected four days after GnRH-induced ovulation. Ampulla and isthmus transcriptome was obtained by RNA-seq, regional gene expression was assessed by qPCR, and PGR and ERa protein distribution was evaluated by immunohistochemistry. There was a greater abundance of PGR and ERa in the oviduct of LF-LCL animals thus indicating a greater availability of receptors and possibly sex steroids stimulated signaling in both regions. Transcriptomic profiles indicated a series of genes associated with functional characteristics of the oviduct that are regulated by the periovulatory sex steroid milieu and that potentially affect oviductal receptivity and early embryo development. They include tissue morphology changes (extra cellular matrix remodeling), cellular changes (proliferation), and secretion changes (growth factors, ions and metal transporters), and were enriched for the genes with increased expression in the LF-LCL group. In conclusion, differences in the periovulatory sex steroid milieu lead to different oviductal gene expression profiles that could modify the oviductal environment to affect embryo survival and development

Clinical and Complementary Diagnosis of Sinus Neoplasia in Horses

Background: Sinus neoplasms are reported as low frequency in horses. Its clinical characteristics are often nonspecific, depend on complementary methods for diagnosis, and when diagnosed, generally they are already advanced, limiting therapeutic possibilities. The objective of this case series was to detail clinical aspects and complementary exams for sinus neoplasms for early diagnosis, comparing them with the literature.Cases: Four horses were treated at the College of Veterinary Medicine and Animal Science of the University of São Paulo, with different clinical signs and previous diagnoses, which when evaluated by respiratory endoscopy, radiography, oral cavity evaluation and histopathological exams, revealed the diagnosis of anaplastic carcinoma, poorly differentiated carcinoma, ossifying fibroma and lymphoma. Previous diagnosis, based mainly on clinical signs, were sinusitis secondary to apical infection, ethmoidal hematoma, sinus cyst and sinusitis secondary to periapical disease. The cases of anaplastic carcinoma (case 1) and lymphoma (case 4) presented with an advanced degree of the disease with involvement and destruction of paranasal structures and adjacent tissues, in addition to a poor general condition, which made surgical treatment impossible and led to euthanasia. In these cases, previous treatment was made to apical infection and periodontal disease with secondary sinusitis, but negative evolution led to suspicion of neoplasia, confirmed by histopathological exam of incisional biopsy of the mass in oral cavity. Benign ossifying fibroma (case 3) presented with progressive respiratory difficulty due to occlusion of the lumen of some nasal meatus and radiographic signs of invasion and deformation of the adjacent bones, it was submitted for surgical excision and there was no recurrence until hospital discharge. The poorly differentiated carcinoma (case 2) was a multilobulate neoformation in the ethmoidal region, similar to ethmoid hematoma in endoscopic and radiographic evaluation, it was submitted for excision and two sessions of electrochemotherapy with bleomycin, associated with administration of piroxicam, which obtained a good result until the period of 1 year after discharge.Discussion: It was found that many characteristics are common with these types of neoplasms, and the clinical signs, such as nasal secretions, airway obstruction, increased facial volume, severe alterations in oral cavity, although unspecific, suggest the differential diagnosis for neoplasms. It is important to differentiate from other diseases noting the evolution and growth of these tumors, like in cases 1 and 4, especially the growth to internal tissues, using complementary methods described here, like endoscopic and radiographic examination. Late identification or even manipulation of neoplasms, without proper diagnosis, leads to a few prognoses regarding life. When it is possible to perform surgical excision, complementary methods are important to guide the procedure, and definitive diagnosis is made through histopathologic exam and some need immunohistochemistry analysis. Cases 2 and 3 had surgical access, were submitted to excision and treatment with good results, both with close monitoring in first months and prolonged quality of life. These results highlight the importance of complementary methods for early diagnosis, correct intervention and monitoring of evolution.Keywords: sinus neoplasia, paranasal sinus, tumor, head, horses

The CÃOCER Project: an Educational Approach for Cancer Prevention in Pet Animals

The CÃOCER projetct started four years ago and comprised several intra and extramural activities with the purpose to aware people of the importance of cancer in animals, focusing mainly in cancer prevention. With this intent, we provided a series of lectures for undergraduates and professionals of veterinary medicine. In addition, we participated in science fairs and exhibitions to disseminate the knowledge regarding cancer prevention for the population from Pirassununga, a city in the country of São Paulo State. Also, we conducted the first campaign of cancer prevention in animals named I Cãopanha of Cancer Prevention in Animals in which preventive physical clinical examinations were done on animals and educational material on cancer prevention was distributed. At last, it was also an objective of this work to perform a statistical survey of cancer cases in animals of Pirassununga. Some undergraduate students of Veterinary Medicine collaborated in this project with or without scholarships. Past four years since its inception, we believe that the project has achievedits initial goals and now our intention is to expand it. For this, we created a website(www.projetocaocer.com.br) that provides articles on cancer prevention in animals.We also wanted to increase the extension of the projetct activities, associating it withthe Brazilian Association of Veterinary Oncology (ABROVET) and the Support ResearchCenter in Veterinary Oncology from USP.O projeto CÃOCER há quatro anos compreende atividades intra e extramurais à USP com a finalidade de conscientizar a população sobre a importância dos cânceres em animais, focando em sua prevenção. Foram realizados ciclos de palestras para alunos de graduação em Medicina Veterinária e profissionais. Participamos de feiras de ciência e exposições para divulgação à população de Pirassununga, e realizamos a I Cãopanha de Prevenção de Câncer em Animais em que foram feitos exames clínicos nos animais e distribuído material educativo referente à prevenção de câncer. Por fim, ainda objetivou-se realizar um levantamento estatístico da casuística de animais portadores de neoplasias atendidos no Hospital Veterinário da FMVZ no campus de Pirassununga da USP. Colaboraram com este projeto diversos alunos de graduação em Medicina Veterinária da FZEA-USP e alunos do ensino médio que obtiveram bolsas pelo Programa de Pré-Iniciação Científica da USP. Passados quatro anos desdeseu início, o projeto atingiu seus objetivos iniciais e agora o intuito é expandi-lo. Para isso criamos um site (www.projetocaocer.com.br) que disponibiliza artigos sobreprevenção do câncer em animais. Pretendemos, também, aumentar as atividades extensionistas do projeto, junto à Associação Brasileira de Oncologia Veterinária e ao NAP de Oncologia Veterinária da Pró-Reitoria de Pesquisa da USP