Transplantation of human immature dental pulp stem cell in dogs with chronic spinal cord injury

Purpose: To investigate the therapeutic potential of human immature dental pulp stem cells in the treatment of chronic spinal cord injury in dogs. Methods: Three dogs of different breeds with chronic SCI were presented as animal clinical cases. Human immature dental pulp stem cells were injected at three points into the spinal cord, and the animals were evaluated by limb function and magnetic resonance imaging (MRI) pre and post-operative. Results: There was significant improvement from the limb function evaluated by Olby Scale, though it was not supported by the imaging data provided by MRI and clinical sign and evaluation. Conclusion: Human dental pulp stem cell therapy presents promising clinical results in dogs with chronic spinal cord injuries, if used in association with physical therapy.(AU)

Rabbit olfactory stem cells. Isolation protocol and characterization

PURPOSE:To describe a new technique for isolation of a mesenchymal stem cells (MSCs) population from the olfactory mucosa in rabbits.METHODS:Olfactory stem cells (OSCs) were retrieved from under the cribriform plate of the Ethmoid bone. Several assays were accomplished to characterize the cell population and attest its viability in vitro. The cells were submitted to flow cytometry with the antibodies CD34, CD45, CD73, CD79, CD90 and CD105 and also they were induced to differentiate in three lineages. Functional evaluation involved analysis of in vitro growth behavior, colony forming unit like fibroblasts (CFU-f) and cryopreservation response. Further transduction with Green Fluorescent Protein (GFP) was also performed.RESULTS:The OSCs showed mesenchymal features, as positive response to CD34, CD73 and CD90 antibodies and plasticity. Additionally, these cells have high proliferated rate, and they could be cultured through many passages and kept the ability to proliferate and differentiate after cryopreservation. The positive response to the transduction signalizes the possibility of cellular tracking in vivo. This is a desirable feature in case those cells are used for pre-clinical trials.CONCLUSION:The cells harvested were mesenchymal stem cells and the technique described is therefore efficient for rabbit olfactory stem cells isolation.(AU)

Isolation, expansion, differentiation and growth kinetics essay in mesenchymal stem cells culture from the bone marrow of collared peccaries (Tayassu tajacu)

Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing. Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS. Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. [...](AU)

Isolation, expansion, differentiation and growth kinetics essay in mesenchymal stem cells culture from the bone marrow of collared peccaries (Tayassu tajacu)

Background: There are few studies on stem cell isolation in wild animals that provide isolation and culture protocols of these cells in vitro. Among the wild species studied, we present the collared peccary (Tayassu tajacu) as a model with potential to obtain and use MSC in preclinical studies. These animals are phylogenetically close to the domestic pig, popularly known as peccaries and found naturally in South America, Central America and the South of the United States. The aim of the present study was to establish a protocol for the isolation, in vitro cell expansion, differentiation and assessment of the stromal MSC growth curve before and after thawing. Materials, Methods & Results: Mesenchymal stem cells (MSC) from collared peccary bone marrow (Tayassu tajacu) were isolated and expanded by centrifuge in Ficoll® solution and cultured in DMEM® High Glucose medium. The culture was assessed by assays of colony forming units CFU-F and growth curve by saturation (GCS). Cultures in the third passage, with 70% confluence, were replicated at 105 cells/mL concentration in the culture media to induce osteogenic cell differentiation and adipogenic cell differentiation, respectively. The MSC were frozen in nitrogen for 40 days, thawed and re-assessed for cell viability and GCS. Discussion: The bone marrow collected presented high mononuclear cellularity, with a mean variability of 94.5% and 60.83 ± 4.27 UFC were identified in the samples and cells with fibroblast-like-cell morphology were observed. When they were expanded, the mean cell viability was 95%, the mean cell concentration obtained was 233.31 ± 20.04 cells per 25cm2 bottle and the culture reached the growth plateau in GCS between the 13th and 16th day. The osteoblastic cell differentiation assay showed after 18 days, morphology similar to osteoblasts, with irregular cytoplasm limits, cell prolongation formation and flattened appearance. [...]

Collared Pecary (Tayassu tajacu) as a new model of renal ischemic injury induced by clamping the renal artery

PURPOSE:The use of the collared peccary as an experimental model for ischemic nephropathy.METHODS:A total of 12 collared peccary (Tayassu tajacu) was used and ischemic nephropathy was induced in six of these animals that constituted the experimental group (G1) while the other six formed the control group (G2). Ischemic nephropathy was induced surgically by partial occlusion of the left renal artery. The disease course was assessed by hematological tests, serum chemistry, urinalysis, ultrasound (US) and doppler ultrasound function of the renal artery before induction, and at five, 10, 15 and 20 days after surgery. Twenty days after the occlusion, unilateral nephrectomy and histopathological examination were performed to assess renal morphology.RESULTS:Statistical analysis by Fischer's test showed a significant difference (p<0.05) between the control group and the experimental group. The histopathological examination showed glomerular, tubular and interstitial lesions. In the experimental group, 83.3% (5 /6) showed moderate renal lesions and only 16.7% (1/6) were classified with no lesions. The ultrasound examination of the right kidney presented statistical difference between day 5 and day 10 post occlusion.CONCLUSION:The collared peccary as a good experimental model for ischemic renal disease, because it could be manipulated during the research time without death, with health conditions that permit any subsequent procedure for disease therapy.(AU)

Isolation, culture and differentiation potential of collared peccary (Tayassu tajacu) adipose-derived stem cells

Background: The understanding of cell biology and the isolation of mesenchymal stem cells in wild animals show prospects for conducting pre-clinical trials in these unconventional animals. The collared peccary (Tayassu tajacu) are suiforms that belong to the Artiodáctyla order, Tayassuidae family and Tayassu genus. They adapt easily to captivity conditions that favors their commercial rearing and is an alternative for biodiversity conservation. To evaluate the collared peccary (Tayassu tajacu) as a potential animal model for the isolation of mesenchymal progenitor cells, cell culture and cell differentiation protocols. Materials, Methods & Results: To perform this research we used four collared peccaries (Tayassu tajacu) from the Nucleus of Study and Preservation of Wild Animals (IBAMA/PI No . 02/08-618) from Federal University of Piauí (UFPI). Adipose tissue fragments were collected from the dorsocervical region and dissociated mechanically in laboratory. The material was placed in an incubator containing CO2 - 95% at 37C and the cultures were expanded to fifth passage, evalluating cell concentration and viability. The culture medium alfa-MEM supplemented was changed every three days. The cell kinetics was evaluated in triplicate using growth curve performed during ten days, plating the initial concentration of 5 x 104 cells/mL per well in P3 six-well culture plate...(AU)

Isolation, culture and differentiation potential of collared peccary (Tayassu tajacu) adipose-derived stem cells

Background: The understanding of cell biology and the isolation of mesenchymal stem cells in wild animals show prospects for conducting pre-clinical trials in these unconventional animals. The collared peccary (Tayassu tajacu) are suiforms that belong to the Artiodáctyla order, Tayassuidae family and Tayassu genus. They adapt easily to captivity conditions that favors their commercial rearing and is an alternative for biodiversity conservation. To evaluate the collared peccary (Tayassu tajacu) as a potential animal model for the isolation of mesenchymal progenitor cells, cell culture and cell differentiation protocols. Materials, Methods & Results: To perform this research we used four collared peccaries (Tayassu tajacu) from the Nucleus of Study and Preservation of Wild Animals (IBAMA/PI No . 02/08-618) from Federal University of Piauí (UFPI). Adipose tissue fragments were collected from the dorsocervical region and dissociated mechanically in laboratory. The material was placed in an incubator containing CO2 - 95% at 37C and the cultures were expanded to fifth passage, evalluating cell concentration and viability. The culture medium alfa-MEM supplemented was changed every three days. The cell kinetics was evaluated in triplicate using growth curve performed during ten days, plating the initial concentration of 5 x 104 cells/mL per well in P3 six-well culture plate...

Isolation and characterization of mesenchymal progenitors derived from the bone marrow of goats native from northeastern Brazil

PURPOSE: To characterize bone marrow progenitors cells grown in vitro, using native goats from northeastern Brazil as animal model. METHODS: Ten northeastern Brazil native goats of both genders were used from the Piauí Federal University Agricultural Science Center's (UFPI) - Goat Farming Sector. Bone marrow aspirates where taken from the tibial ridge and seeded on culture plates for isolation, expansion and Flow Cytometry (expression markers - Oct-3/4, PCNA, Ck-Pan, Vimentina, Nanog). RESULTS: Progenitor cells showed colonies characterized by the presence of cell pellets with fibroblastoid morphology. Cell confluence was taken after 14 days culture and the non-adherent mononuclear cell progressive reduction. After the first passage, 94.36% cell viability was observed, starting from 4.6 x 106 cell/mL initially seeded. Cells that went through flow cytometry showed positive expression for Oct-3/4, PCNA, Ck-Pan, Vimentina, and Nanog. CONCLUSIONS: Bone marrow progenitor isolated of native goats from northeastern Brazil showed expression markers also seen in embryonic stem cells (Oct-3/4, Nanog), markers of cell proliferation (PCNA) and markers for mesenchymal cells (Vimentina and Ck-pan), which associated to morphological and culture growth features, suggest the existence of a mesenchymal stem cell (MSC) population in the goat bone marrow stromal cells studied.(AU)

Protocols for obtainment and isolation of two mesenchymal stem cell sources in sheep / Protocolos para obtenção e isolamento de células tronco mesenquimais de duas fontes em ovinos

PURPOSE: To evaluate different protocols to isolate stem cells from ovine umbilical cord blood and adipose tissue. METHODS: There were used 5 samples of umbilical blood and 5 samples of perirenal adipose tissue from 10 female sheep. All the samples were obtained through surgery, to harvest aseptic samples. There were used 3 protocols for obtainment and culture of umbilical cord blood stem cells and 4 protocols for ovine adipose tissue stem cells. RESULTS: It was possible to observe only one successful protocol for the obtainment of umbilical cord blood stem cells. When analyzing the techniques used to obtain adipose tissue stem cells, only one of the methods was effective as well. Through colony forming unit assay, there were obtained 58 colonies of cells after seven days in culture. Flow citometry tests revealed the cells were positive to CD44 and exhibited negative reaction to CD38, CD45, CD41/61. These cells showed a growth curve with very well defined phases LOG, LAG and PLATEAU. This phases are typically seem in mesenchymal stem cells growth curves. CONCLUSIONS: The isolation and culture of mesenchymal stem cells from ovine umbilical cord blood are complex and request more detailed assays. Stem cells from fat tissue sheep showed mesenchymal characteristics, according to their cell growth curve, ability to origin colonies of fibroblastoid cells and positive reactivity with the antibody CD44 by flow citometry.(AU)

OBJETIVO: Testar diferentes protocolos para o isolamento de células tronco a partir de sangue de cordão umbilical e tecido adiposo de ovinos. MÉTODOS: Foram utilizadas cinco amostras de sangue de cordão umbilical e cinco amostras de tecido adiposo perirrenal de 10 fêmeas de ovelha. A coleta das amostras foi realizada através de procedimento cirúrgico para coleta do material de forma mais asséptica possível. Foram realizados três protocolos de isolamento e cultivo das células-tronco do cordão umbilical e quatro protocolos para o isolamento e cultivo das células-tronco de gordura de ovinos RESULTADOS: Somente um dos protocolos utilizados para o isolamento das células-tronco de cordão umbilical foi efetivo. Dos quatro protocolos utilizados para isolamento das células-tronco de gordura, da mesma forma, apenas um obteve sucesso. Foi realizado o ensaio de unidades formadoras de colônias destas células, sendo contadas 58 colônias ao final de sete dias. Na citometria de fluxo essas células mostraram-se positivas para CD44 e negativas para CD38, CD45, CD41/61. Estas células apresentaram curva de crescimento com fases de LOG, LAG e PLATEAU bem definidas, características das curvas de crescimento das células-tronco de origem mesenquimal. CONCLUSÕES: O isolamento e cultivo das células-tronco mesenquimais do cordão umbilical de ovinos é de difícil realização, exigindo maiores ensaios e estudos profundos. Células tronco do tecido adiposo de ovelhas demonstraram características mesenquimais, de acordo com a curva de crescimento, habilidade de formação de colônias, células com morfologia fibroblastóide e reação positiva ao anticorpo CD44.(AU)

Successful transplant of mesenchymal stem cells in induced osteonecrosis of the ovine femoral head: preliminary results / Sucesso no transplante de células tronco mesenquimais em ovinos com osteonecrose induzida da cabeça do fêmur: resultados preliminares

PURPOSE: Evaluate the bone tissue recovery following transplantation of ovine mesenchymal stem cells (MSC) from bone marrow and human immature dental-pulp stem cells (hIDPSC) in ovine model of induced osteonecrosis of femoral head (ONFH). METHODS: Eight sheep were divided in three experimental groups. First group was composed by four animals with ONFH induced by ethanol through central decompression (CD), for control group without any treatment. The second and third group were compose by two animals, six weeks after ONFH induction received transplantation of heterologous ovine MSC (CD + oMSC), and hIDPSC (CD + hIDPSC), respectively. In both experiments the cells were transplanted without application of any type of immunosupression protocol. RESULTS: Our data indicate that both cell types used in experiments were able to proliferate within injured site providing bone tissue recovery. The histological results obtained from CD+hIDPSC suggested that the bone regeneration in such animals was better than that observed in CD animals. CONCLUSION: Mesenchymal stem cell transplant in induced ovine osteonecrosis of femoral head by central decompression technique is safe, and apparently favors bone regeneration of damaged tissues.(AU)

OBJETIVO: Verificar os efeitos das células-tronco mesenquimais da medula óssea de ovinos e da polpa dentária imatura humana em ovinos com osteonecrose induzida, da cabeça do fêmur. MÉTODOS: Oito ovelhas foram distribuídas em três grupos experimentais. O primeiro grupo foi composto por quatro animais com osteonecrose da cabeça do fêmur induzida por etanol através da descompressão central, que não receberam nenhum tratamento. O segundo e o terceiro grupo, cada um composto por dois animais, receberam transplante heterólogo de células tronco mesenquimais de ovinos e polpa dentária imatura humana seis semanas após a indução da osteonecrose da cabeça do fêmur, respectivamente. Em ambos os grupos experimentais as células foram transplantadas sem o uso de drogas imunossupressoras. RESULTADOS: Os achados demonstram que as células-tronco mesenquimais injetadas na cabeça do fêmur se encontravam viáveis após o transplante no novo sítio e proliferaram em pouco tempo. Os dados histológicos sugerem que a regeneração óssea nos animais transplantados com polpa dentária imatura humana foi mais rápida do que nos animais submetidos somente a descompressão central. CONCLUSÃO: O transplante de células tronco mesenquimais na osteonecrose da cabeça do fêmur induzida em ovinos através da técnica de descompressão central é um procedimento seguro, e aparentemente favorece a regeneração óssea de tecidos lesados.(AU)

Terapia celular com células-tronco em coelhos com lesão medular induzida e em cães com lesão medular crônica espontânea / Stem cell therapy in rabbits with induced spinal cord injury and in dogs with spontaneous chronic spinal cord injury

A regeneração de danos às células nervosas do sistema nervoso central (SNC) é limitada devido aos fatores intrínsecos ao próprio nicho celular. A cicatriz glial, a liberação de inibidores de crescimento axonal, e a ação fagocítica dos astrócitos são fatores que limitam essa regeneração, coibindo os processos de reestruturação axonal. As lesões medulares causam um grande impacto na vida de seus portadores e de seus familiares. O uso das células tronco surgiu como uma alternativa viável ao tratamento das lesões medulares em humanos e animais. No entanto, apesar das tentativas terapêuticas em animais apresentarem resultados animadores, os ensaios clínicos realizados em humanos carecem de estudos mais aprofundados, e ainda não apresentaram resultados satisfatórios. O presente trabalho propôs a avaliação clínica do uso de células tronco da polpa dentária humana em cães com lesão medular crônica, e o uso das células tronco do epitélio olfatório de coelhos em coelhos com lesão medular induzida por hemissecção medular dorsal. Três cães foram submetidos à intervenção cirúrgica e avaliados através de teste comportamental e de ressonância magnética por imagem. Seis coelhos foram submetidos à técnica cirúrgica de hemissecção medular dorsal, e foram transplantados com células tronco do epitélio olfatório de coelhos e avaliados através de histologia e imunohistoquímica. As células-tronco do epitélio olfatório de coelhos foram rastreadas na medula espinhal dos animais 20 dias após a realização da terapia celular, confirmando o sucesso do transplante. A expressão da proteína fluorescente verde (GFP) através da imunohistoquímica comprovam o sucesso do transplante celular. O caso clínico canino um apresentou uma melhora clínica acentuada, pulando de um escore 4 para um escore de 8 em um curto espaço de tempo, mas não apresentou melhora nos exames de ressonância magnética. O caso clínico canino dois apresentou melhora clínica, embora a ressonância magnética mostrasse uma piora na situação da medula espinhal. E nosso caso clínico canino três apresentou mínima melhora clínica, sem alteração na imagem pré e pós operatória. Os dados obtidos com este estudo comprovam que a terapia celular com células tronco na lesão medular podem ser benéficas, no entanto ainda não representam uma cura para as lesões medulares crônicas

The regeneration of damaged nerve cells of the central nervous system (CNS) is limited due to factors intrinsic to the own cell niche. The glial scar, the release of axonal growth inhibitors, and the phagocytic action of astrocytes are factors that limit this regeneration, deterring axonal restructuring. The spinal cord injuries cause a great impact on the lives of sufferers and their families. The use of stem cells has emerged as a viable alternative to the treatment of spinal cord injuries in humans and animals. However, despite attempts therapies in animals show encouraging results, clinical trials performed in humans require further study, and have not had satisfactory results. This paper proposed the clinical use of stem cells from human dental pulp in dogs with chronic spinal cord injury, and the use of stem cells from the rabbit olfactory epithelium in rabbits with spinal cord injury induced by dorsal hemisection. Three dogs underwent surgery and evaluated by behavioral test and magnetic resonance imaging. Six rabbits underwent surgical technique for spinal cord dorsal hemisection and were transplanted with stem cells from the olfactory epithelium of rabbits and evaluated by histology and immunohistochemistry. Stem cells of the olfactory epithelium of rabbits were traced in the spinal cord of animals 20 days after the completion of cell therapy, confirming the success of transplantation. Green fluorescent protein (GFP) tracked by immunohistochemical evidence the success of cell transplantation. The canine clinical case one presented a marked clinical improvement, jumping from a score of 4 to a score of 8 in a short time, but there was no improvement in magnetic resonance imaging. The canine clinical case two showed clinical improvement, although the MRI showed a worsening situation in the spinal cord. And our canine clinical case 3 showed minimal clinical improvement, no change in image pre-and postoperatively. The data obtained from this study show that cell therapy with stem cells in spinal cord injury may be beneficial, but still not a cure for chronic spinal cord injuries

Transplante heterólogo de células-tronco mesenquimais em ovinos com osteonecrose da cabeça do fêmur induzida / Heterologous Transplant of Bone Marrow Mesenchymal Stem Cells in Ovine With Osteonecrosis of The Femoral Head Induced

A Osteonecrose da cabeça do fêmur (ONCF) é uma patologia degenerativa que pode levar ao colapso da cabeça do fêmur tendo como conseqüência a necessidade de substituição da articulação coxofemoral. O objetivo principal no tratamento da ONCF é preservar a articulação coxofemoral e não substituí-la. Estudos realizados em pacientes humanos com ONCF demonstraram que os níveis de atividade e o número de células mesenquimais nos compartimentos hematopoiéticos e estromais da medula óssea estão deprimidos. Estes achados indicam a possibilidade da utilização de células-tronco mesenquimais (MSC) nas lesões necróticas da cabeça do fêmur. O presente trabalho propõe a utilização da terapia celular com MSC da medula óssea de ovinos e polpa dentária humana em 8 ovinos com ONCF induzida pela injeção intra-óssea de etanol, além da avaliação da reestruturação óssea através da microscopia de luz, e rastreamento das células transplantadas. Dois animais foram eutanasiados com 6 semanas após a indução da lesão e foram transplantadas as células-tronco mesenquimais em 4 animais, e realizada a descompressão central em 2 outros animais. Com dez semanas foram eutanasiados os animais restantes. O sucesso do transplante da medula óssea foi confirmado pela expressão do gene repórter LacZ na cabeça do fêmur dos animais transplantados macroscopicamente. A análise histológica evidenciou maior processo regenerativo na cabeça do fêmur do animal experimental submetido à terapia com células-tronco da polpa dentária imatura humana

The osteonecrosis of femoral head (ONFH) is a degenerative disease that can lead to the collapse of the femoral head and necessity of total hip replacement. The main objective in the treatment of ONFH is to preserve the hip and not replace it. Studies in human patients with ONFH demonstrated that levels of activity and the number of mesenchymal cells in the hematopoietic and bone marrow stromal compartments are depressed. These findings indicate the possibility of the use of mesenchymal stem cells (MSC) in necrotic lesions on the head of the femur. This research proposes the use of cell therapy with MSC from ovine bone marrow and immature human dental pulp stem cell in ovine ONCF induced by intraosseous injection of ethanol, in addition, the assessment of bone restructuring by light microscopy and tracking of the transplanted cells. Two animals were euthanasied with 6 weeks after the induction of injury, and were transplanted mesenchymal stem cells in 4 animals, and held the central decompression in 2 other animals. After ten weeks were euthanasied the remaining animals. The successful of the bone marrow transplantation was confirmed by the macroscopic expression of the LacZ gene reporter in the femoral head of animals transplanted. The histological analysis showed an advanced regenerative process in the femoral head in the animal using human immature dental pulp stem cell

Contaminação por enteroparasitas em amostras de alfaces (Lactuca sativa) de hortas produtoras de verduras da ilha de São Luís, MA / Contamination for parasites in samples of lettuces (Lactuca sativa) of producing of vegetables of the island of São Luís, MA

Um grande número de enfermidades entéricas são veiculadas através de verduras contaminadas. Este estudo teve como objetivo a verificação da contaminação por enteroparasitas em amostras de alfaces (Lactuca sativa) provenientes de hortas da ilha de São Luís, MA. Do total de 60 amostras analisadas, 96,6 por cento apresentaram-se contaminadas. Foram identificados ovos de ancilostomídeos, Ascaris lumbricoides, Enterobius vermiculares, Hymenolepis sp., Taenia sp., oocistos de coccídeo, cistos de Entamoeba sp. Também se detectou ácaros, ovos de ácaro, fragmentos de ácaros e artrópode, piolho, hifas de fungo, larvas de nematóides. Conclui-se que, as hortaliças já saem das hortas contaminadas e podem veicular enteroparasitas para a população.(AU)

A great number of enteric diseases are transmitted by the ingestion of contaminated vegetables. The objective of this study was to evaluate the contamination by parasites in lettuce produced in kitchen gardens from São Luís lsland, Maranhão. Lt was observed that 96,6% of the 60 samples was contaminated. The following parasites were identified: eggs of aneilostomid, Ascaris lumbricoides, Enterobius vermiculares, Hymenolepis sp., Taenia sp., coceidea oocystis, cystis of Entamoeba coli, Entamoeba histolytica, Entamoeba sp. Arthropod and mite fragments, mites and eggsmites, louse, fungus hyphas, nematode larvae were also detected. We concluded that lettuce analyzed are contaminated in the kitchen gardens and may transmit intestinal parasites. (AU)