Simone Passos Bianchi

Genetic variability and phospholipase production of Malassezia pachydermatis isolated from dogs with diverse grades of skin lesions

Little detailed information is available on the association of Malassezia pachydermatis genotypes and the extent of skin damage that they cause. In the present study, isolates of M. pachydermatis, recovered from the skin of healthy dogs and dogs with dermatitis in Brazil, were characterized on the basis of partial sequencing of the large subunit (LSU), first internal transcribed spacer (ITS-1) and chitin synthase 2 gene (chs-2). The determination of phospholipase production was also included in the investigations. The severity of lesions and hyperpigmentation of dogs with skin disease were evaluated. For each locus, two main sequence types were designated as genotypes A and C. Two other minor sequence types (A2(I)-C2(I)) were also recorded and defined for the ITS-1. Genotype A isolates were the most prevalent, being recovered from healthy and diseased animals. No significant difference was detected among genotypes or ITS-1 sequence types and grades of skin damage or hyperpigmentation in the dogs with skin lesions. The number of M. pachydermatis isolates that produced phospholipase was statistically higher for diseased dogs than for strains found in healthy animals. The present study reveals that multiple genetic variants of M. pachydermatis occur in dogs and that the distribution patterns of particular genotypes on the skin of dogs in Brazil might be related to environmental and ecological factors which maintain distinctive genotype assemblages in specific geographical areas.

Detection of Pneumocystis in lungs of bats from Brazil by PCR amplification

Pneumocystis has been isolated from a wide range of unrelated mammalian hosts, including humans, domestic and wild animals. It has been demonstrated that the genome of Pneumocystis of one host differs markedly from that of other hosts. Also, variation in the chromosome and DNA sequence of Pneumocystis within a single host species has been observed. Since information about the occurrence and nature of infections in wild animals is still limited, the objective of this work was to detect the presence of Pneumocystis sp. in lungs of bats from two states from Brazil by Nested-PCR amplification. The bats, captured in caves and in urban areas, were obtained from the Program of Rabies Control of two States in Brazil, Mato Grosso and Rio Grande do Sul, located in the Mid-Western and Southern regions of the country, respectively. DNAs were extracted from 102 lung tissues and screened for Pneumocystis by nested PCR at the mtLSU rRNA gene and small subunit of mitochondrial ribosomal RNA (mtSSU rRNA). Gene amplification was performed using the mtLSU rRNA, the primer set pAZ102H - pAZ102E and pAZ102X - pAZY, and the mtSSU rRNA primer set pAZ102 10FRI - pAZ102 10R-RI and pAZ102 13RI - pAZ102 14RI. The most frequent bats were Tadarida brasiliensis (25), Desmodus rotundus (20), and Nyctinomops laticaudatus (19). Pneumocystis was more prevalent in the species Nyctinomops laticaudatus (26.3% = 5/19), Tadarida brasiliensis (24% = 6/25), and Desmodus rotundus (20% = 4/20). Besides these species, Pneumocystis also was detected in lungs from Molossus molossus (1/11, 9.1%), Artibeus fimbriatus (1/1, 100%), Sturnira lilium (1/3, 33.3%), Myotis levis (2/3, 66.7%)and Diphylla ecaudata (1/2, 50%). PCR products which could indicate the presence of Pneumocystis (21.56%) were identified in DNA samples obtained from 8 out of 16 classified species from both states (5 bats were not identified). This is the first report of detection of Pneumocystis in bats from Brazil.

Tumoral Thrombus in Cranial Mesenteric Artery Secondary to Intestinal Leiomyosarcoma

Background: Leiomyosarcoma is a non-encapsulated tumor, belonging to the group of soft tissue sarcomas and often invasive. This histological type comprises the most aggressive solid tumors and, because they respond poorly to therapy, are considered the worst prognosis. The dissemination of metastatic cells can occur mainly hematologically to the liver, lungs and bones or by direct invasion, but metastases to blood vessels are rare and surgery is the treatment of choice in most cases. This paper aims to report a rare case of tumor thrombus in the cranial mesenteric artery secondary to an intestinal leiomyosarcoma. Case: A 9-year-old female S.R.D dog presenting inappetence, evolving to anorexia, weight loss, anemia and vomiting for approximately 15 days was referred to the Hospital of Veterinary Clinics of the Federal University of Rio Grande do Sul. Previous therapy at another clinic with three blood transfusions did not promote reversal of anemia. Ultrasonography revealed a nodule in the small intestine of seven cm in diameter and in the hemogram a hypochromic macrocytic anemia. Thoracic radiography did not reveal metastases. After a new blood transfusion, the patient was referred to an exploratory laparotomy, where the nodule was identified in the small intestine, which was cyanotic and two nodules in the cranial mesenteric artery, partially obstructing the local blood flow. Resection and intestinal anastomosis were performed from the final portion of the duodenum to the cecum, with ligation of the mesenteric artery and the nodules were sent for histopathological analysis. The animal presented a gradual improvement of the condition, with the introduction of liquid feeding on the second postoperative day. After these days, he presented hypochromic macrocytic anemia, with presence of Howell-Jolly corpuscle, spherocytes and icteric plasma, compatible with hemolytic anemia. Treatment with prednisone was started, with a worsening of the condition. The patient had generalized jaundice and died five days after surgery. The animal was referred to the necropsy in which it showed abdominal fluid of slightly reddish color, adherence of viscera (intestine, stomach and omentum). In the surgical area (enterectomy) there was purulent secretion on the points, the large intestine presented ecchymosis and petechiae on the mucosa. Histological examinations of the nodule and thrombus revealed malignant mesenchymal neoplasia formed by spindle cells with elongated nuclei, abundant eosinophilic cytoplasm and moderate anaplasia, interspersed by connective tissue, with areas of necrosis and neutrophilic infiltrate. The final diagnostic was intestinal leiomyosarcoma with mesenteric vessel metastasis and septicemia. Discussion: Because the tumor thrombus is located in the mesenteric artery, resection of a large part of the small intestine was necessary because this artery promotes the irrigation of this region. When this procedure is performed, a common complication is short bowel syndrome, necessitating a specific nutritional therapy. The animal probably developed hemolytic anemia by the successive blood transfusions that it underwent, causing in a worsening of the anemia and developing jaundice, not responding to the instituted therapy. In addition, there were rupture of the surgery points, extravasating fluid into the abdominal cavity, triggering a septicemia resulting in the death of the patient. Keywords: intestinal tumor, neoplasia, metastasis.

Células-tronco mesenquimais aplicadas nas fases inflamatória e proliferativa da cicatrização de feridas cutâneas

Wound healing is a process that requires the interaction of various cells in the dermis and epidermis. The aim of this study was to evaluate the action of ADSCs in the treatment of acute wounds in order to understand if application time of the cells results in a difference in healing the first seven days of injury. The stem cells were isolated from adipose tissue of C57BL / 6 mice GFP +. Thus, we used 49 mice C57BL / 6 divided into four groups: Group I (GI / control, n=14); Group II (GII; n=14): ADSCs injected to the d0; Group III (GIII; n=14): ADSCs injected on the 3rd day, and Group IV (GIV; n=7): ADSCs injected day 5(d5). Clinical evaluations were performed on days 0, 3, 5 and 7 and the histopathology on days 5 and 7. In the proposed methodology, the use of ADSCs increased vascularization, formation of granulation tissue, collagen deposition and increases the number of hair follicles in just seven days of evaluation. In addition, the time of application of the cells did not affect significant differences in the inflammatory and the proliferative phase of wound healing skin.