Biocontrol agents in the management of Meloidogyne incognita in tomato / Agentes de biocontrole no manejo de Meloidogyne incognita em tomateiro

Recognized as prominent among the principal pests of tomato crop, the root-knot nematode (Meloidogyne incognita) has generated much study with regards to control methods. This study aimed to assess the efficiency of biological products in decreasing the M. incognita population in tomato. Four experiments were conducted under greenhouse conditions. Two of these experiments had the purpose to determine the efficiency of NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), and the other two to assess the efficiency of other different biological products in M. incognita management. Tomato plants of the cultivar Santa Cruz Kada Giant were inoculated using 2000 eggs and J2 of M. incognita. In the first two experiments doses corresponding to 0, 4, 6, 8 and 10kg ha-1 of NemOut were applied after inoculation. In the second study, the treatments consisted of the use of biological products containing different other biocontrol agents. Assessments were made at 45 and 65 days after inoculation (DAI) to evaluate plant height, fresh shoot and fresh root weigh, gall index and egg mass index, population density and reproduction factor (RF). The NemOut doses reduced the M. incognita population density and RF at 45 DAI. The agent Pochonia chlamydosporia showed greater efficiency in controlling M. incognita on tomato. Biological products and the different doses of NemOut had no influence on the tomato plant development.

O nematoide das galhas (Meloidogyne incognita) é relatado como uma das principais pragas da cultura do tomate, sendo importante o estudo de alternativas para o seu manejo. O presente estudo objetivou avaliar a eficiência de produtos biológicos na redução da população de M. incognita em tomate. Foram conduzidos quatro experimentos em casa de vegetação, dois visando avaliar a eficiência de doses de NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), e outros dois para avaliar a eficiência de diferentes produtos biológicos no controle de M. incognita. Plantas de tomate Santa Cruz Kada Gigante foram inoculadas com 2000 ovos e J2 de M. incognita. No primeiro estudo foram utilizadas doses equivalentes a 0, 4, 6, 8 e 10kg ha-1 de NemOut, aplicadas após a inoculação. No segundo estudo os tratamentos constituíram de produtos biológicos com diferentes agentes de biocontrole. As avaliações foram realizadas 45 e 65 dias após a inoculação (DAI), determinando-se altura de plantas, massa fresca da parte aérea e da raiz, índice de galhas e de massas de ovos, densidade populacional e fator de reprodução (FR). Doses crescentes de NemOut reduziram a densidade populacional e o FR de M. incognita, aos 45 DAI. O agente Pochonia chlamydosporia mostrou-se mais eficiente no controle de M. incognita no tomateiro. Os produtos biológicos e as diferentes doses de NemOut não afetaram o desenvolvimento das plantas de tomate.

Biocontrol agents in the management of Meloidogyne incognita in tomato / Agentes de biocontrole no manejo de Meloidogyne incognita em tomateiro

Recognized as prominent among the principal pests of tomato crop, the root-knot nematode (Meloidogyne incognita) has generated much study with regards to control methods. This study aimed to assess the efficiency of biological products in decreasing the M. incognita population in tomato. Four experiments were conducted under greenhouse conditions. Two of these experiments had the purpose to determine the efficiency of NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), and the other two to assess the efficiency of other different biological products in M. incognita management. Tomato plants of the cultivar Santa Cruz Kada Giant were inoculated using 2000 eggs and J2 of M. incognita. In the first two experiments doses corresponding to 0, 4, 6, 8 and 10kg ha-1 of NemOut were applied after inoculation. In the second study, the treatments consisted of the use of biological products containing different other biocontrol agents. Assessments were made at 45 and 65 days after inoculation (DAI) to evaluate plant height, fresh shoot and fresh root weigh, gall index and egg mass index, population density and reproduction factor (RF). The NemOut doses reduced the M. incognita population density and RF at 45 DAI. The agent Pochonia chlamydosporia showed greater efficiency in controlling M. incognita on tomato. Biological products and the different doses of NemOut had no influence on the tomato plant development.(AU)

O nematoide das galhas (Meloidogyne incognita) é relatado como uma das principais pragas da cultura do tomate, sendo importante o estudo de alternativas para o seu manejo. O presente estudo objetivou avaliar a eficiência de produtos biológicos na redução da população de M. incognita em tomate. Foram conduzidos quatro experimentos em casa de vegetação, dois visando avaliar a eficiência de doses de NemOut (Bacillus subtilis + B. licheniformis + Trichoderma longibrachiatum), e outros dois para avaliar a eficiência de diferentes produtos biológicos no controle de M. incognita. Plantas de tomate Santa Cruz Kada Gigante foram inoculadas com 2000 ovos e J2 de M. incognita. No primeiro estudo foram utilizadas doses equivalentes a 0, 4, 6, 8 e 10kg ha-1 de NemOut, aplicadas após a inoculação. No segundo estudo os tratamentos constituíram de produtos biológicos com diferentes agentes de biocontrole. As avaliações foram realizadas 45 e 65 dias após a inoculação (DAI), determinando-se altura de plantas, massa fresca da parte aérea e da raiz, índice de galhas e de massas de ovos, densidade populacional e fator de reprodução (FR). Doses crescentes de NemOut reduziram a densidade populacional e o FR de M. incognita, aos 45 DAI. O agente Pochonia chlamydosporia mostrou-se mais eficiente no controle de M. incognita no tomateiro. Os produtos biológicos e as diferentes doses de NemOut não afetaram o desenvolvimento das plantas de tomate.(AU)

Adenocarcinoma papilar de seio nasal em cão: abordagem histomorfológica, imunofenotípica e terapêutica / Papillary Adenocarcinoma of Nasal Sinus in a Dog: Approach Histomorphologic, Immunophenotypic, and Therapeutic

Background: Sinus adenocarcinomas of papillary pattern are neoplasms rarely described in dogs and most cases are characterized by slow growth, local invasion and rare cases of metastasis. Their characteristics are presence of glandular structures that may have papillary pattern, tubulo-papillary, acinar and mixed pattern. Approximately 80% of primary nasal tumors are malignant and adenocarcinomas are the most frequently histology type. This paper reports a case of papillary adenocarcinoma of nasal sinus in the dog, emphasizing the use of histochemical and immunohistochemical techniques to aiding for appropriate therapy. Case: A 14-year-old male Poodle dog, presenting seropurulent nasal discharge, frequent sneezing and an enlargement in the nasal region, was submitted to surgical excision of a tumor. Macroscopically, was observed fragment coated with soft hairy skin and shear blackish color, and some fragments associated with more firm tissues like cartilage and bone tissue. Tumor specimens were collected, fixed in 10% neutral buffered formalin solution and embedded in paraffin. Afterwards, 4 µm histological sections were obtained and stained with HE. For immunohistochemical analysis a biotin-peroxidase system was used and secondary antibodies were identifi ed using Advance HRP. CKAE1/AE3, CK34ßE12, SMA, E-cadherin, Cox-2, Ki-67 and Her-2 expression were evaluated. For CK AE1AE3 antibody a qualitative method was used and neoplasia were classified as negative (-) or positive (+) and for CK34ßE12 was used semiquantitative evaluation and neoplasms were classified as negative (-), positive with focal staining (+) and positive with diffuse staining (++). The graduation for HER-2 expression was (+) for 4 (++++), were 4 (++++) was the most intensity membrane staining. For Cox-2 the system used was semiquantitative and the distribution was scored by estimating the percentage of tumor cells staining. Intensity was estimated on a scale from 0 (absent) to 12 (strong). The proliferative index was calculated by counting nuclei positive for Ki-67 (anti-MIB-1) in a total of 1000 neoplastic cells. Microscopically, was observed a proliferation of epithelial cells in papillary arrangement with delicate connective axis of support and areas of stromal invasion, and the tumor received the diagnosis of papillary adenocarcinoma of nasal sinus. Immunohistochemical analysis revealed strong staining for Cox-2 (score 12), CKAE1/AE3 positivity and CK34ßE12 (+ + +) and negative for Her-2 (+). Loss of E-cadherin expression was observed and SMA allowed the visualization of areas of basal membrane rupture. The rate of proliferation was evaluated by means of the expression of MIB-1 was high (16%). Chemotherapy was performed with carboplatin and gemcitabine. Discussion: Sinus carcinomas in general, despite their slow growth, are locally invasive and cause extensive destruction of adjacent tissues. In this case, the immunohistochemical analysis presented positive expression of CK AE1/AE3 and CK34ßE12, indicating epithelial histogenesis with basal phenotype. Strong intensity of COX-2 expression was observed, indicating a poor prognostic for the patient. The loss of expression of E-caderin, areas of invasive tumors and high rate of cell proliferation, demonstrated the aggressive character of papillary adenocarcinoma nasal sinus. Surgery with chemotherapy- associated was effective and tolerant treatment. Diagnosis was based on histopathological and immunohistochemical analysis. Correct characterization of this neoplasm associated with the appropriate therapy aided in prognosis and in the improvement of the animal's quality of life. The patient's overall survival has been evaluated since the surgical procedure up to this date and is over 18 months.

Diagnóstico diferencial de tumor estromal gastrointestinal canino / Differential diagnosis of canine gastrointestinal stromal tumor

Background: Stromal Grastrointestinal Tumors (GIST) are rare mesenchymal neoplasms that affect the wall of the gastrointestinal tract (GIT). Histologically this tumor shows a spindle, epithelioid or mixed aspect and routine hematoxylin and eosin (HE) staining does not allow adequate differentiation from other mesenquimal neoplasms. Histochemical and imunohistochemical techniques are useful tools in this differentiation. The aim of this study was to report a case of canine GIST, emphasizing the use of histochemical and immunohistochemical techniques to differentiate it from other histomorfologically similar mesenchymal neoplasms. Case: A ten-year-old male Pinscher dog was submitted to surgical excision of a tumor in the pyloric region of stomach. The dog was euthanized 90 days after surgery. Tumor specimens were collected, fixed in 10% neutral buffered formalin solution and embedded in paraffin. Afterwards, 4 µm histological sections were obtained and stained with HE and Gomori's Tricrome (GT). For immunohistochemical analysis a biotin-peroxidase system was used and secondary antibodies were identified using Advance HRP. Cytokeratin (CK) AE1/AE3, c-KIT, vimentin, Ki-67, S-100 and Smooth Muscle Alfa Actin (SMA) expression were evaluated. For vimentin, CKAE1AE3, SMA, S100 and c-KIT antibodies, a semiquantitative method was used and neoplasms were classified as negative (-), positive with focal or multifocal staining (+) and positive with diffuse staining (++). The proliferative index was calculated by counting nuclei positive for Ki-67 (anti-MIB-1) in a total of 1000 neoplastic cells. Macroscopically, the stomach fragment revealed an intramural nodule covered by mucosa and with a slightly lobulated surface. Microscopically, the neoplastic mass was located in the submucous layer of stomach and composed by a spindle cell proliferation forming intersecting fascicles. Neoplastic cells revealed vacuolated eosinophilic cytoplasm and multiple conspicuous nucleoli. Absence of chromatic affinity for GT was observed in this tumor. Immunohistochemical analysis revealed intense and diffuse cytoplasmic staining for vimentin (++), moderate and diffuse staining for c-KIT (++), moderate and multifocal staining for S-100 (+) and negative staining (-) for SMA and CK AE1/AE3. The proliferation index was 6.8%. Discussion: GISTs are mesenquimal neoplasms that affect the GIT. The spindle aspect of this tumor is similar to other mesenquimal tumors of the GIT. A mesenquimal appearance associated with epithelioid areas was observed in this case report. Mesenquimal histogenesis was confirmed by positivity for vimentin and negativity for cytokeratin. GT evaluation revealed absence of muscular and collagenous components within the neoplasm, discarding the diagnosis of leiomiosarcoma or fibrosarcoma. The absence of smooth muscle tumor cells was confirmed by immunohistochemical negativity for SMA. In addition, multifocal and diffuse cytoplasmic imunostaining was observed for S-100 and c-KIT, respectively. According the literature, human and canine GISTs are positive for c-KIT and negative for S-100. The S-100 expression described in this study, has been reported in humans GISTs and is probably associated with neural differentiation. Therefore, histomorfological and immunohistochemical findings allowed a definitive diagnosis of canine GIST, differentiating the tumour from other mesenchymal neoplasms of the GIT.