Origin and distribution pattern of pelvic limb nerves of a Bengal tiger (Panthera tigris tigris).

The Bengal tiger (Panthera tigris tigris) is a species belonging to the Felidae family. In Argentina, tigers are currently only found in captivity. The longevity of individual animals in human-controlled environments depends on proper management and practices that prioritize animal welfare. Regular veterinary care is essential to maintain optimal health conditions. Professionals must have a comprehensive understanding of the anatomy and physiology of tigers to effectively perform medical procedures and administer treatments. The study described in the text focuses on the trajectory and distribution of nerves in the pelvic limb of a Bengal tiger specimen, providing detailed dissection findings. The results revealed that the lumbosacral plexus is formed from the ventral rami of the LIV, LV, LVI, LVII, SI, SII and SIII nerves. Among the observations to highlight is the great development of the nerves N. cutaneus femoris lateralis and N. cutaneus femoris caudalis some differences were observed in the distribution of the N. femoralis and N. obturatorius; the N. ischiadicus, together with its division into the fibularis communis and tibialis nerves, showed the same configuration observed in other cats. Finally, it was observed that the nerves N. gluteus cranialis and N. gluteus caudalis also originated from the truncus lumbosacralis. The similarities and differences with studies carried out on other cats are relevant and provide anatomical data for medical procedures in the Bengal tiger.

Embryonic-placental relationship in Lagostomus maximus as compared to other hystricognath rodents and eutherian mammals.

Reproductive specializations in caviomorphs (infraorder Hystricognathi), are very peculiar within the Order Rodentia. These include long gestations, the birth of offspring with an extreme degree of precociality, and short lactation periods. This study describes the embryo-placental relationship of viable implantation sites (IS) of the plains viscacha, Lagostomus maximus, after 46 post-coital days. The observations recorded in this study are comparatively discussed with those of other hystricognaths and eutherians. At this stage, the embryo resembles that of other eutherians. At this time of embryo development, the placenta exhibits a size, shape, and organization similar to that it will have in its mature state. Besides, the subplacenta is already highly folded. These characteristics are adequate to sustain the development of future precocial offspring. The mesoplacenta, a structure present in other hystricognaths and related to uterine regeneration is described for the first time in this species. This detailed description of the placental and embryonic structure contributes to the knowledge of the reproductive and developmental biology of the viscacha, as well as that of hystricognaths. These characteristics will allow testing other hypotheses related to the morphology and physiology of the placenta and subplacenta, and their relationship with the growth and development of precocial offspring in Hystricognathi.

Spontaneous embryonic death in plains viscacha (Lagostomus maximus - Rodentia), a species with unique reproductive characteristics.

Spontaneous embryonic death is a conserved reproductive event in Eutherians. The macro and microscopic characteristics of this type of death are similar between the different taxa. However, in the hystricomorphic rodent plains viscacha (Lagostomus maximus) is exceptional in terms of massiveness (80% embryonic resorption). In this species, of the 10-12 implantation sites (IS) (half in each uterine horn), only the caudal embryos will survive, resorbing the cranial and intermediate IS. We hypothesize that uterine structural variations in L. maximus restrict growth and promote embryo death, with the consequent loss of placental homeostasis in the cranial and middle IS. In this study, different studies (ultrasonography, macroscopy and microscopy) were carried out to analyze different aspects of the intermediate gestation of L. maximus (46 days postcoitus). Ultrasonographic studies revealed that the cranial and middle IS (IS-1, IS-2, and IS-3) had no recognizable embryonic and placental structures as compared to the caudal implantation sites (IS-4). Macroscopically, the areas corresponding to the embryos in the cranial and middle IS were occupied by a necrotic black semi-fluid mass. Moreover, the placenta in these IS was undifferentiated. However, in the caudal IS both the embryo and its placenta were distinguishable. Using histological and immunohistochemical techniques, it was observed that the placentas of IS-1, IS-2 and IS-3 were disorganized and showed hemorrhage, inflammatory infiltration containing neutrophils, macrophages, mast cells and foreign body giant cells, apoptotic trophoblast, and a layer of collagen fibers and fibroblasts that circumscribed each of these IS. In contrast, the placenta of the caudal IS showed an organized maternal-embryonic interface. The characteristics observed in IS in resorption of viscachas in intermediate gestation show that, regardless of gestation time, embryonic death has a similar macro and microscopic morphological pattern among eutherians with invasive placentation. However, the massiveness and sectorization of embryonic death in the plains viscacha make the species a unique model for the study of this reproductive event.

Placentation and embryo death in the plains viscacha (Lagostomus maximus).

Caviomorpha are an exceptional group among rodents due to their extended gestational period and the delivery of precocial offspring. Among them, Lagostomus maximus is characterized by its polyovulation, polyembryony, and the highest embryonic death known in mammals. Its chorioallantoic placenta is hemomonochorial, an ancestral character among rodents. It resembles more the human placenta than the murine models. As in all caviomophs, the chorioallantoic placenta is divided in a main placenta and a subplacenta. The former is organized in labyrinth lobes surrounded by trophospongium, as in most caviomorphs. The giant cells (more numerous than in other caviomorphs) near the decidua could be related to invasiveness. During placentation of L. maximus, uterine natural killer cells are found. These cells have been related to invasiveness and remodeling of blood vessels in Mus musculus and Homo sapiens, although in other caviomorphs are not frequently found. In L. maximus, the placenta develops in all conceptuses (5-6 per uterine horn). Necrosis was observed in each implantation site at day 70 post-coitum, except in that closest to the vagina in each horn. This process of embryo death followed by resorption begins at day 26-30 post-coitum. Recently, we found variations in the percentage of blood vessel and uterine gland areas that could explain the regional differences in embryo survival. The characteristics of the placenta and implantation of L. maximus are important to stablish a unique model for studying placentation as well as early embryonic death, of interest for human and veterinary medicine.

Analysis of glycoconjugates and morphological characterization of the descending colon and rectum of the plains viscacha, Lagostomus maximus.

Herbivores exhibit specializations at the intestinal level that facilitate the bacterial fermentation. The available information on the digestive physiology of Lagostomus maximus makes this rodent an interesting model to evaluate morpho-functional adaptations to herbivory. The general objective of this work was centered on the study of the morphology and histochemistry of the descending colon and rectum of L. maximus. To do so, a comparative analysis of the morphology, ultrastructure and glycosylation pattern of both anatomical regions was carried out. Histochemical results revealed that in both sectors of the large intestine, there are goblet cells with different glycosylation pattern within a morphologically homogeneous cell population. The main difference between both intestinal segments lay in the fact that the most distal region of the large intestine showed a greater proportion of sialomucins, characterized by being slightly O-acetylated. Further specific differences were revealed by lectin histochemistry. These data allowed to perform a functional interpretation of the cell types and secreted substances, thus contributing to a better understanding of the role of mucins in the intestinal tract functioning.

Reactivity of microglia and astrocytes after an excitotoxic injury induced by kainic acid in the rat spinal cord.

After injury of the nervous system glial cells react according to the stimuli by modifying their morphology and function. Glia activation was reported in different kainic acid (KA)-induced neurodegeneration models. Here, we describe glial morphometric changes occurring in an excitotoxic KA-induced cervical spinal cord injury model. Concomitant degenerative and apoptotic processes are also reported. Male rats injected at the spinal cord C5 segment either with KA or saline were euthanized at post-injection (PI) days 1, 2, 3 or 7. Anti-IBA-1 and anti-GFAP antibodies were used to identify microglia and activated astrocytes, respectively, and to morphometrically characterized them. Fluoro-Jade B staining and TUNEL reaction were used to determine neuronal and glial degeneration and apoptosis. KA-injected group showed a significant increase in microglia number at the ipsilateral side by PI day 3. Different microglia reactive phenotypes were observed. Reactive microglia was still present by PI day 7. Astrocytes in KA-injected group showed a biphasic increase in number at PI days 1 and 3. Degenerative and apoptotic events were only observed in KA-injected animals, increasing mainly by PI day 1. Understanding the compromise of glia in different neurodegenerative processes may help to define possible common or specific therapeutic approaches directed towards neurorestorative strategies.

Ghrelin Recruits Specific Subsets of Dopamine and GABA Neurons of Different Ventral Tegmental Area Sub-nuclei.

Ghrelin is a stomach-derived hormone that regulates rewarding behaviors and reinforcement by acting on the ventral tegmental area (VTA). The VTA is a complex midbrain structure mainly comprised of dopamine (DA) and gamma-aminobutiric acid (GABA) neurons that are distributed in several VTA sub-nuclei. Here, we investigated the neuroanatomical distribution and chemical phenotype of ghrelin-responsive neurons within the VTA. In wild-type mice, we found that: (1) ghrelin binding cells are present in most VTA sub-nuclei but not in its main target, the nucleus accumbens (Acb); (2) systemically injected ghrelin increases food intake but does neither affect locomotor activity nor the levels of the marker of neuronal activation c-Fos in the VTA sub-nuclei; (3) centrally injected ghrelin increases food intake, locomotor activity and c-Fos levels in non-DA neurons of all VTA sub-nuclei; (4) intra-VTA-injected ghrelin increases food intake, locomotor activity and c-Fos levels in non-DA neurons of all VTA sub-nuclei; (5) both centrally and intra-VTA-injected ghrelin increase c-Fos levels in DA neurons of the parabrachial pigmented VTA sub-nucleus. In genetically modified mice in which a subset of GABA neurons expresses the red fluorescent protein tdTomato, we found that centrally injected ghrelin increases c-Fos levels in GABA neurons of the interfascicular VTA sub-nucleus. These results suggest that ghrelin can recruit specific subsets of VTA neurons in order to modulate food intake and locomotor activity.

Lechiguana em bovinos: aspectos patogênicos / Lechiguana in cattle: pathogenic aspects

Dois bovinos foram infectados com larvas de Dermatobia hominis na região escapular esquerda, e o local de infecção de um dos bovinos foi inoculado com Mannheimia granulomatis. Aos sete, 14, 21 e 28 dias, foram feitas biópsias cirúrgicas dos locais de inoculação das larvas para posterior estudo histológico, imuno-histoquímico e avaliação por microscopia eletrônica de transmissão. No bovino infectado somente com as larvas, as lesões histológicas se caracterizaram por dermatite eosinofílica, infiltrado de células mononucleares e proliferação de tecido conjuntivo. A inoculação simultânea de Dermatobia hominis e Mannheimia granulomatis provocou adicionalmente linfangite e microabscessos eosinofílicos, similares aos observados em casos espontâneos da enfermidade. Na amostra coletada aos 28 dias após a infecção bacteriana, foi observada, tanto na imuno-histoquímica, como por meio da microscopia eletrônica, a presença de corpos bacterianos aparentemente intactos no interior de fagossomas macrofágicos, sugerindo que a bactéria permanece dentro dos macrófagos até que algum fator desconhecido desencadeie o desenvolvimento da enfermidade. A Mannheimia granulomatis foi isolada de larvas coletadas aos 42 dias após a inoculação, demonstrando que a bactéria pode sobreviver por algum tempo na larva de Dermatobia hominis. Os dados do trabalho indicam que as larvas de Dermatobia hominis podem atuar como um fator contribuinte para o desenvolvimento da paniculite fibrogranulomatosa proliferativa.

Two cattle were infected at the left scapula with larvae of the Dermatobia hominis, while one of them was infected with the Mannheimia granulomatis. Biopsies of the lesions were taken at the 7, 14, 21 and 28 days p.i. for subsequent histopatological, immunohistochemical and electron microscopical studies. In the only animal infected with larvae, eosinophilic dermatitis, infiltration of mononuclear cells and proliferation of connective tissue characterized the histological lesions. The simultaneous inoculation of Dermatobia hominis and Mannheimia granulomatis caused, besides the characteristic lesions of the parasitosis, linfangitis and eosinophilic microabscesses, similar changes to those observed in spontaneous cases of the disease. In the samples taken after 28 days of the bacterial infection it was observed, in both immunohistochemestry and by electronic microscopy the presence of apparently intact bacterial bodies inside macrophages fagosomes, suggesting that the bacteria is inside the macrophages until some unknown factors trigger the development of the illness. Mannheimia granulomatis was isolated from larvae gathered 42 days after inoculation demonstrating that the bacteria can survive inside the larva Dermatobia hominis. Data from the study indicate that the larvae of Dermatobia hominis may act as a contributing factor to the development of proliferative fibrogranulomatous panniculitis.

Decrease in PTEN and increase in Akt expression and neuron size in aged rat spinal cord.

PTEN is a tumor suppressor gene known to play an important role in the regulation of cell size. In this study we compared PTEN expression in the spinal cord of young (5 months old) vs. aged (32 months old) female rats and correlated them with alterations in neuron size and morphology in the same animals. Total and phosphorylated PTEN (pPTEN) as well as its downstream target phosphorylated Akt (pAkt) were assessed by Western blotting. Spinal cord neurons were morphometrically characterized. Total PTEN, pPTEN and total Akt expression were significantly higher in young rats than in aged animals. Expression of pAkt was stronger in aged animals. A significant increase in neuronal size was observed in large motoneurons of aged as compared with young rats. Our data show that in the spinal cord of rats, neuronal PTEN expression diminishes with advanced age while neuronal size increases. These results suggest that in the spinal cord, an age-related reduction in PTEN and increase of pAkt expression may be involved in the progressive enlargement of neurons.

Impact of very old age on the expression of cervical spinal cord cell markers in rats.

Aging is a process associated with both anatomical changes and loss of expression of some cell markers. Intermediate filaments are known to impart mechanical stability to cells and tissues. Some of them are present in different cell populations of the central nervous system. In order to explore the impact of extreme age we immunohistochemically characterized the changes in intermediate filaments and other cellular markers present in cells populating the gray matter cervical spinal cord of very old rats (28 months) taking young (5 months) counterparts as a reference. The spinal cord weight of the senile animals (12.6+/-1.1 g) was significantly higher (P<0.001) than that of the young animals (8.4+/-1.1 g). Spinal cord length also increased significantly (P<0.05) with age (7.9+/-0.3 cm vs. 8.28+/-0.1 cm for young and senile, respectively). An increase in both neurofilament staining area and density was observed in senile rats in comparison to young animals. A significant (P<0.05) age-related increment in the mean area of the cervical segments was observed. Vimentin expression in the ependymal zone decreased in area and intensity during aging. Our data show that there are some significant changes in the morphological and histochemical patterns of the cervical spinal cord in senile rats. However, they do not necessarily represent a pathologic situation and may rather reflect plastic reorganization.

Presence of binucleate neurons in the spinal cord of young and senile rats.

The presence of binucleate cells constitutes a normal feature of some animal tissues but is rare in the normal brain and has not been documented in the spinal cord. We assessed different segments of the rat spinal cord in order to determine the frequency and distribution of binucleate neurons in this structure as well as the impact of aging on this neuronal population. Young (4-5 months) and senile (32 months) female Sprague-Dawley rats were used. Sections from cervical, thoracic and lumbar segments were histochemically and immunohistochemically (NeuN) stained and the frequency and distribution of binucleate neurons was determined by manual counting. The frequency of binucleate neurons in all of the analysed segments was comparable between young and senile animals. Binucleate neurons were particularly frequent in the C5 and C6 segments. The overall distribution of binucleate neurons in the different laminae assessed was, Lm-III = 19%; Lm-VI = 17%; Lm-VII = 39%; LmVIII = 8%; Lm-IX = 11%; Lm-X = 6%, and was comparable between young and senile rats. We conclude that binucleate neurons occur as a normal feature of the rat spinal cord and that their frequency and distribution does not change with aging.

Loss of NeuN immunoreactivity in rat spinal cord neurons during aging.

A morphologic study of the impact of aging on neuron marker expression was performed in different segments of the rat spinal cord. Spinal cord specimens from young (5 months), middle-aged (12 months) and senile (32 months) female rats were assessed. We found a complete loss of neuron-specific nuclear protein (NeuN) immunoreactivity in cervical, thoracic and lumbar segments of the senile animals whereas neuron-specific enolase (NSE) immunoreactivity was comparable in young and senile rats. These findings in otherwise morphologically well preserved spinal cord neurons are of interest and reveal that NeuN may not be a reliable marker to identify neurons in the spinal cord of aging rats.

Intoxicação experimental com frutos de Melia azedarach (Meliaceae) em suínos / Experimental poisoning by fruits of Melia azedarach (Meliaceae) in pigs

Frutos maduros, triturados, de Melia azedarach foram administrados misturados à ração a 8 suínos em doses únicas de 5-20g/kg. Diarréia passageira foi o único sinal nos 2 suínos que consumiram 5g/kg. Os sinais clínicos observados nos animais que ingeriram doses únicas de 10, 15 e 20g/kg caracterizaram-se por incoordenação, tremores musculares, dificuldade para manter-se de pé, relutância para levantar-se, decúbito esternal e hipotermia. Os 2 suínos que ingeriram 20g/kg morreram. Macroscopicamente observou-se congestão do intestino delgado e da mucosa do estômago. As alterações histológicas caracterizaram-se por discreta a moderada necrose do tecido linfático, necrose dos músculos esqueléticos, congestão e necrose da mucosa gástrica e do intestino delgado e alterações degenerativas moderadas no fígado e rim. Os níveis séricos de AST e CPK aumentaram após a ingestão do tóxico. Observaram-se valores altos de CPK nos animais que morreram, evidenciando a mionecrose. Os resultados obtidos demonstram que, para suínos, a ingestão de altas doses de frutos de Melia azedarach induz ao aparecimento de sinais clínicos da intoxicação e causa lesões em diversos órgãos, inclusive a morte.

Bone changes caused by experimental Solanum malacoxylon poisoning in rabbits

The aim of this study was to describe the bone changes observed after a daily oral administration of the calcinogenic plant Solanum malacoxylon (syn. S. glaucophyllum) (Sm) during 9 days. The Sm-poisoned rabbits had an increase of bone resorption in the endosteal surface of the cortical zone and also in the surface covered by osteoblasts of the primary and secondary spongiosa of the trabecular bone compartment. Moreover, the epiphyseal growth plates in long bones appeared narrower than in the control rabbits, with reduction of the proliferative and hyperthrophic chondrocyte zones. The electron microscopic study revealed a significant decrease of proteoglycans in the hyperthrophic chondrocyte zone evidenced by a significant reduction of rutenium red positive granules in the poisoned rabbit. Altogether, these data suggest that cell differentiation may play a pivotal role in the pathogenesis of Sm-induced bone lesions.

Morphometry of cervical segments grey matter in the male rat spinal cord.

The cervical portion of the spinal cord is an area frequently affected by alterations of medical and veterinary importance. Since there is scarce quantitative anatomical data on this region, we undertook a morphometric study of the grey matter of all segments of the rat cervical spinal cord of male rats in order to generate reference patterns to be used in future experimental studies. Using image analysis software, the total spinal cord length and grey and white matter area of each segment was recorded. The morphometric characteristics of the neurones populating the laminae of the grey matter of the cervical segments was also recorded. Neurones were classified into small, medium and large sizes for each lamina and statistically compared. The present data fill an anatomical information gap by providing quantitative data about the normal anatomical features of the rat cervical cord. The anatomical data found could be used to better understand the physiological relevance of that region in the rat.

Impact of very old age on hypothalamic dopaminergic neurons in the female rat: a morphometric study.

Dopaminergic neurons of the A(12) (tuberoinfundibular dopaminergic system) and A(14) (periventricular dopaminergic system) hypothalamic areas exert a tonic inhibitory control of prolactin secretion. Tuberoinfundibular dopaminergic system neuron function is known to decline during aging in rats, but little is known about the impact of extreme age on neuron number and morphology in the two systems. We morphometrically assessed the neurons of the tuberoinfundibular dopaminergic system and the periventricular dopaminergic system in female rats 6 (young, Y), 24 (old, O), and 30-32 (senescent, S) months old. Serial coronal sections of fixed hypothalami were immunohistochemically labeled for tyrosine hydroxylase, and immunoreactive perikarya from the A(12) and A(14) areas were quantitatively characterized and compared among the three age groups. Radioimmunoassay was used to measure serum prolactin. The number of A(12) tyrosine hydroxylase-immunoreactive perikarya showed a steady decline with age, whereas the number of A(14) tyrosine hydroxylase-immunoreactive perikarya remained stable from young to old age but showed a sharp drop in the senescent rats. In the old rats, tyrosine hydroxylase-immunoreactive neuronal area (A(12) = 135.37 and A(14) = 158.79 microm(2)) was significantly higher than that of young (A(12) = 72.56 and A(14) = 99.7 microm(2)) and senescent animals (A(12) = 95.5 and A(14) = 106.5 microm(2)). Densitometric assessment of median eminence tyrosine hydroxylase immunoreactivity revealed a steady age-related reduction of tyrosine hydroxylase content in the median eminence. Serum prolactin levels increased steadily with age. We conclude that, in the female rat, aging brings about a progressive loss of both tuberoinfundibular dopaminergic system and periventricular dopaminergic system neurons, which becomes more conspicuous at extreme ages.

Immunohistochemical detection of Tritrichomonas foetus in experimentally infected mice

A necessidade de aumentar o conhecimento da patogenia da tricomoníase genital bovina (BGT) conduziu ao uso de modelos experimentais alternativos como o camundongo. Näo obstante, é necessário elucidar a dinâmica da infecçäo nesta espécie e avaliar as diferentes fases da colonizaçäo e evoluçäo das alterações patológicas. A imunohistoquímica (IHQ) oferece vantagens sobre as técnicas histoquímicas de rotina para a observaçäo do protozoário em tecidos, detritos celulares e dentro de macrófagos. O objetivo do presente trabalho foi demonstrar pelo uso de uma técnica de IHQ a presença de Tritrichomonas foetus no sistema reprodutivo de camundongos infectados. Camundongos BALB/c fêmeas foram infectados pela via intravaginal, com uma suspensäo de T. foetus, na fase de estro, detectado com citologia exfoliativa vaginal. Depois de 10 semanas, os animais foram sacrificados; útero e vagina forma fixados e processados para histologia. Alguns cortes foram corados com HE. O restante dos cortes foi processado para IHQ. Foi usado um soro policlonal imunoadsorvido anti-T. foetus. A técnica de avidina-biotina (HistoMouse, ZymedÖ) foi empregada. Os estudos histopatológicos mostraram uma dilataçäo das glândulas uterinas, presença de macrófagos no lúmen do órgäo e parte interna das glândulas endometriais. T. foetus näo foi identificado por esse método. A IHQ permitiu identificar as mesmas lesões observadas e a presença do protozoário no endométrio, nas glândulas endometriais, no lúmen uterino e dentro de neutrófilos e macrófagos. O estudo citológico em lâminas coradas por IHQ, mostrou T. foetus aderido a células epiteliais, ou dentro de macrófagos. Esta técnica demonstra ser uma ferramenta útil para o estudo da patogenia da tricomoníase genital bovina (BGT) utilizando-se o camundongo como modelo experimental.

Detection of illegal estrogen administration through immunohistochemical markers in the bovine prostate

A imunodeteccäo de marcadores celulares foi avaliada em amostras prostáticas de bovinos com hiperplasia ou hiperplasia-metaplasia epiteliais, induzidas por estrógenos administrados ilegalmente e em próstatas de bovinos inoculados com dietilstilbestrol (DES). A técnica de recuperacäo antigênica permitiu o uso de tecidos fixados em formalina, por longos períodos. Foram utilizados os anticorpos monoclonais K8.12, anti-vimentina e anti-desmina para determinaçäo de células basais coradas/epitélio hiperplásico-metaplásico, células do estroma e células musculares, respectivamente. As alterações tissulares observadas nos casos de campo e nos experimentais foram semelhantes, através do que se concluiu que houve administraçäo ilegal de estrógenos. O teste imuno-histoquímico com esses marcadores específicos foi útil ao exame histológico da próstata, uma vez que a análise das imagens permite maior e melhor quantificaçäo das alterações observadas. Os testes bioquímicos, entretanto, säo necessários para uma avaliaçäo mais precisa

Modificación del método original para la determinación de proteínas / Modification of the original method for protein determination

Se presenta una modificación de la técnica original para la determinación cuantitativa de proteínas, propuesta por Lowry y colaboradores. Se modificó la concentración de los reactivos empleados para dicha determinación, lo que permite una reducción en la cantidad de los mismos. Asimismo, se estandarizó en 10 min el tiempo de incubación con cada uno de los reactivos empleados, reduciendo, de esta manera, a sólo 20 min el tiempo para la lectura de la reacción. La densidad óptica se leyó con una longitud de onda de 660 nm. Esta técnica ha demostrado ser tan sensible para la determinación proteica de los líquidos corporales analizados, como la originalmente propuesta por Lowry y col

Modificación del método original para la determinación de proteínas / Modification of the original method for protein determination

Se presenta una modificación de la técnica original para la determinación cuantitativa de proteínas, propuesta por Lowry y colaboradores. Se modificó la concentración de los reactivos empleados para dicha determinación, lo que permite una reducción en la cantidad de los mismos. Asimismo, se estandarizó en 10 min el tiempo de incubación con cada uno de los reactivos empleados, reduciendo, de esta manera, a sólo 20 min el tiempo para la lectura de la reacción. La densidad óptica se leyó con una longitud de onda de 660 nm. Esta técnica ha demostrado ser tan sensible para la determinación proteica de los líquidos corporales analizados, como la originalmente propuesta por Lowry y col (AU)