Lectinhistoquímica del granuloma inducido por el bacilo de Calmette Guérin en Piaractus mesopotamicus

Objective: This study was conducted to evaluate, by means of lectinhistochemistry (LHC), the expression of carbohydrates in granulomas induced by the bacillus Calmette-Guerin (BCG) in muscle tissue of Piaractus mesopotamicus after 33 days. Material and methods: Histological sections with 3 µm thick were incubated with the following lectins: WGA (Wheat germ agglutinin), DBA (Dolichos biflorus agglutinin) and HPA (Helix pomatia agglutinin), and the results were evaluated by light microscopy. Results: Acid fast bacilli were stained by Ziehl Neelsen (ZN) and strong labeled by WGA in the cytoplasm of macrophages. Labeling with DBA was intense in fibroblasts and weak in macrophages. On the other hand, HPA binding was stronger in macrophages, especially in those that were in close contact with epithelioid cells, without evidence of binding to fibroblasts. The epithelioid cells were not labeled by the used lectins, but they were identified by Hematoxilin-Eosin (HE). The lectins labeled specific type saccharides in glycoproteins, as N-acetylglucosamine present in bacilli and macrophages, as well as N-acetyl-galactosamine in macrophages. The control group showed no inflammation or lectin binding. Conclusions. This technique may be useful in identifying receptors for WGA, DBA and the HPA lectins in epithelioid granuloma induced by BCG in P. mesopotamicus.Objetivo: El presente estudio fue realizado para evaluar por medio de lectinhistoquímica (LHC), la expresión de carbohidratos en granulomas inducidos por el bacilo de Calmette-Guérin (BCG) en músculo de Piaractus mesopotamicus después de 33 días. Materiales y métodos: Cortes histológicos de 3 µm de grosor fueron incubados con las siguientes lectinas: WGA (Wheat germ aglutinin), DBA (Dolichos biflorus agglutin) y HPA (Helix pomatia agglutinin), y los resultados evaluados por medio de microscopia de luz. Resultados: Bacilos ácido resistentes fueron identificados por la tinción de Ziehl Neelsen(ZN). Se observó un marcaje intenso con WGA en el citoplasma de macrófagos. El marcaje con DBA fue intenso en fibroblastos y débil en macrófagos. Con la lectina HPA el marcaje fue intenso en macrófagos, principalmente en los que estaban en estrecho contacto con las células epitelióides, externamente se observó marcaje débil en fibroblastos. Las células epitelióides no fueron marcadas por las lectinas, pero fueron identificadas con la tinción de Hematoxilina-Eosina (HE). Las lectinas tuvieron un tipo de marcaje específico en algunos monosacáridos, como N-acetilglucosamina presente en los bacilos y en macrófagos, y N-acetilgalactosamina en macrófagos. En el grupo control no fue observada inflamación así como tampoco marcaje con las lectinas. Conclusiones. Esta técnica resultó eficiente en la identificación de receptores para las lectinas WGA, DBA y HPA en el granuloma epitelióide inducido por BCG en P. mesopotamicus.Facultad de Ciencias Veterinaria

Lectinhistoquímica del granuloma inducido por el bacilo de Calmette Guérin en Piaractus mesopotamicus

Objective: This study was conducted to evaluate, by means of lectinhistochemistry (LHC), the expression of carbohydrates in granulomas induced by the bacillus Calmette-Guerin (BCG) in muscle tissue of Piaractus mesopotamicus after 33 days. Material and methods: Histological sections with 3 µm thick were incubated with the following lectins: WGA (Wheat germ agglutinin), DBA (Dolichos biflorus agglutinin) and HPA (Helix pomatia agglutinin), and the results were evaluated by light microscopy. Results: Acid fast bacilli were stained by Ziehl Neelsen (ZN) and strong labeled by WGA in the cytoplasm of macrophages. Labeling with DBA was intense in fibroblasts and weak in macrophages. On the other hand, HPA binding was stronger in macrophages, especially in those that were in close contact with epithelioid cells, without evidence of binding to fibroblasts. The epithelioid cells were not labeled by the used lectins, but they were identified by Hematoxilin-Eosin (HE). The lectins labeled specific type saccharides in glycoproteins, as N-acetylglucosamine present in bacilli and macrophages, as well as N-acetyl-galactosamine in macrophages. The control group showed no inflammation or lectin binding. Conclusions. This technique may be useful in identifying receptors for WGA, DBA and the HPA lectins in epithelioid granuloma induced by BCG in P. mesopotamicus.Objetivo: El presente estudio fue realizado para evaluar por medio de lectinhistoquímica (LHC), la expresión de carbohidratos en granulomas inducidos por el bacilo de Calmette-Guérin (BCG) en músculo de Piaractus mesopotamicus después de 33 días. Materiales y métodos: Cortes histológicos de 3 µm de grosor fueron incubados con las siguientes lectinas: WGA (Wheat germ aglutinin), DBA (Dolichos biflorus agglutin) y HPA (Helix pomatia agglutinin), y los resultados evaluados por medio de microscopia de luz. Resultados: Bacilos ácido resistentes fueron identificados por la tinción de Ziehl Neelsen(ZN). Se observó un marcaje intenso con WGA en el citoplasma de macrófagos. El marcaje con DBA fue intenso en fibroblastos y débil en macrófagos. Con la lectina HPA el marcaje fue intenso en macrófagos, principalmente en los que estaban en estrecho contacto con las células epitelióides, externamente se observó marcaje débil en fibroblastos. Las células epitelióides no fueron marcadas por las lectinas, pero fueron identificadas con la tinción de Hematoxilina-Eosina (HE). Las lectinas tuvieron un tipo de marcaje específico en algunos monosacáridos, como N-acetilglucosamina presente en los bacilos y en macrófagos, y N-acetilgalactosamina en macrófagos. En el grupo control no fue observada inflamación así como tampoco marcaje con las lectinas. Conclusiones. Esta técnica resultó eficiente en la identificación de receptores para las lectinas WGA, DBA y HPA en el granuloma epitelióide inducido por BCG en P. mesopotamicus.Facultad de Ciencias Veterinaria

Inflamación crónica granulomatosa en el pez teleósteo Piaractus mesopotamicus: modelo de estudio histopatológico

Objective. This study evaluated the cell kinetic and formation of granuloma during chronic inflammation induced by Bacillus Calmette-Guérin (BCG) in the skeletal muscle of Piaractus mesopotamicus, as a histopathology model to study innate immunity. Materials and methods. Sixty fish were divided in two groups: BCG-inoculated and non-inoculated fish and the inflammatory response analyzed 3, 7, 14, 21 and 33 days post-inoculation (DPI) by histopathology after hematoxylin-eosin and Ziehl-Neelsen staining. Results. 3 DPI of BCG showed a diffuse inflammatory reaction mostly composed by mononuclear cells. The inflammation continued diffuse 7 DPI initiating the cellular organization surrounding the inoculum and have continued at 14 DPI with discrete presence of epithelioid-like type cells with acidophilic cytoplasm and floppy chromatin. Higher cellular organization (21 DPI) surrounding the granuloma with intense peripheral mononuclear inflammatory infiltrate and nevertheless, an increase in the number of fibroblasts and macrophage-like cells was observed. The inflammatory process became less diffuse 33 DPI with formation of small amount of granuloma surrounded by the same type of reaction found in bigger granuloma. Both the young and old granuloma presented typical characteristic around the inoculum composed by a layer of epithelioid-like type cells, besides macrophages, some lymphocytes and abundant fibroblasts. Conclusions. This study showed the feasibility in the use of pacus to study chronic granulomatous inflammatory response induced by BCG, characterized by changes in the kinetics of inflammatory cells in skeletal muscle classifying as immune-epithelioid type, similar to granulomatous inflammation caused by M. marinum in teleost fish.Objetivo. Este estudio evaluó la cinética celular y la formación de granuloma durante la inflamación crónica inducida por el Bacilo Calmette-Guérin (BCG) en el músculo esquelético de Piaractus mesopotamicus, como modelo histopatológico para estudiar la inmunidad innata. Materiales y métodos. Sesenta peces fueron divididos en dos grupos: peces inoculados con BCG y no inoculados y la respuesta inflamatoria analizada en 3, 7, 14, 21 y 33 días post-inóculo (DPI) por medio del análisis histopatológico y tinciones de hematoxilina-eosina y Ziehl-Neelsen. Resultados. 3 DPI de BCG se observó reacción inflamatoria difusa principalmente formada por infiltrado celular mononuclear. Al 7° DPI la inflamación continuaba difusa con inicio de organización celular alrededor del inoculo, que se observó hasta el 14° DPI con discreta presencia de células de tipo epiteliodes con citoplasma acidófilo y cromatina laxa. Para el 21° DPI se observó alta organización celular alrededor del granuloma con intenso infiltrado mononuclear periférico e incremento en el número de fibroblastos y macrófagos. El proceso inflamatorio se tornó menos difuso a los 33 DPI con formación de pequeños granulomas contenidos dentro de uno más grande. Los granulomas formados más rápidamente así como los formados tardíamente, presentaron características típicas alrededor del inóculo compuesta por una camada de células tipo epitelioides, macrófagos, linfocitos y fibroblastos. Conclusiones. Este estudio mostró la viabilidad del uso del P. mesopotamicus para estudiar la respuesta inflamatoria crónica granulomatosa inducida con BCG, caracterizado por la evolución de la cinética de células inflamatorias en el músculo esquelético clasificándolo como de tipo inmune-epitelioide, similar a la inflamación granulomatosa causada por M. marinum en peces teleósteos

Clinical safety of dichlorvos (45%), cypermethrin (5%) and piperonyl butoxide (25%) administered by spray on the skin of cattle

ABSTRACT Objective. Due to the importance of controlling ectoparasites, associated with the necessity of technical knowledge on the safety of topical treatment with organophosphates, pyrethroids and piperonyl butoxide to the animal organism, this bioassay was carried out to evaluate the clinical safety of the association of dichlorvos (45%) + cypermethrin (5%) + piperonyl butoxide (25%) administered by spray on the skin of cattle, through the study of clinical parameters, biochemical, haematological and behavioral changes. Materials and methods. Sixteen crossbred animals with a mean age of 18 months, males and females grouped into two treatments with eight animals each: T1 (1:800 v/v) and T2 (1:200 v/v). Were collected blood samples at six different times: before treatment (BT), 24, 48, 72, 96 and 192 hours post treatment (HPT). Results. The antiparasitic association administered by spray on the skin did not result in changes in the enzymatic activity of ALT, AST, GGT and ALP, as well as in serum albumin, triglycerides, cholesterol, urea and creatinine, demonstrating the safety of this antiparasitic compound for maintaining hepatic and renal functionality. The erythrocyte, leukocyte and platelet studies showed no changes caused by treatments, and no clinical signs and behavioral changes were observed after treatment. Conclusions. These findings demonstrated good safety margin for spray treatment on the skin with this antiparasitic compound, even when administered at a dilution of 1:200 v/v, which is four times the dose recommended for ectoparasite control

Lectinhistochemical staining of granuloma induced by bacillus Calmette-Guerin in Piaractus mesopotamicus

ABSTRACT Objetive. This study was conducted to evaluate, by means of lectinhistochemistry (LHC), the expression of carbohydrates in granulomas induced by the bacillus Calmette-Guerin (BCG) in muscle tissue of Piaractus mesopotamicus after 33 days. Material and methods. Histological sections with 3 μm thick were incubated with the following lectins :WGA (Wheat germ agglutinin), DBA (Dolichos biflorus agglutinin) and HPA (Helix pomatia agglutinin), and the results were evaluated by light microscopy. Results. Acid fast bacilli were stained by Ziehl Neelsen (ZN) and strong labeled by WGA in the cytoplasm of macrophages. Labeling with DBA was intense in fibroblasts and weak in macrophages. On the other hand, HPA binding was stronger in macrophages, especially in those that were in close contact with epithelioid cells, without evidence of binding to fibroblasts. The epithelioid cells were not labeled by the used lectins, but they were identified by Hematoxilin-Eosin (HE). The lectins labeled specific type saccharides in glycoproteins, as N-acetylglucosamine present in bacilli and macrophages, as well as N-acetyl-galactosamine in macrophages. The control group showed no inflammation or lectin binding. Conclusions. This technique may be useful in identifying receptors for WGA, DBA and the HPA lectins in epithelioid granuloma induced by BCG in P. mesopotamicu

Estreptococos beta-hemolítico en tilapias del Nilo (Oreochromis niloticus) cultivadas en Sullana, Piura - Perú

Objective. This investigation aimed to study the presence of Streptococcus spp. in tilapia (Oreochromis niloticus) from fish farm located in Sullana-Piura, Peru. Materials and methods. 150 fish with clinical signs of streptococcal disease were sampled, and the bacterium isolation was performed on blood agar, correlated to histopathological lesions description and molecular confirmation by real-time PCR. Results. The necropsy revealed exophthalmia, hyphema, congestion and/or haemorrhagic meninges, ascites, splenomegaly, hepatomegaly and diffuse haemorrhagic zones throughout the body. 102 isolated positives (54 tilapias) to Streptococcus spp. were identified in the microbiological analysis (prevalence of 26%), the brain was the organ with the highest percentage of this bacteria (34.31%), and 19 isolates were beta-haemolytic (18.63%) with prevalence of 10.12%. Fish beta-haemolytic streptococci presented epicarditis, perisplenitis and chronic meningitis, panophthalmitis, coagulative necrosis of skeletal muscle and granulomas formation. In the confirmatory test by real-time PCR, any positive tilapia to S. iniae was obtained. The results were analysed using a stochastic simulation of beta distribution using @Risk program uncertainty, reporting an average prevalence of 0.66% in sick tilapias. Conclusions. The analysed fishes were positive to bacteria of the genus Streptococcus, which confirms its presence in the fish farm. However, 19 isolates were beta-haemolytic, and the presence of S. iniae was not positive to the limit prevalence of 2.7% in real-time PCR.Objetivo. Esta investigación objetivó estudiar la presencia de Streptococcus spp. en tilapia (Oreochromis niloticus) de la piscifactoría localizada en Sullana-Piura, Perú. Materiales y Métodos. 150 peces con signos clínicos de la enfermedad estreptocócica fueron analizados, el aislamiento bacteriano se realizó en agar sangre, en correlación con las lesiones histopatológicas y diagnóstico molecular mediante PCR en tiempo real. Resultados. A la necropsia se observó exoftalmia, hifema, congestión y/o meninges hemorrágicas, ascitis, esplenomegalia, hepatomegalia y zonas hemorrágicas difusas en todo el cuerpo. 102 aislados positivos (54) tilapias para Streptococcus spp. fueron identificados en el análisis microbiológico (prevalencia del 26%), el cerebro fue el órgano con el más alto porcentaje del género de esta bacteria (34.31%), 19 aislados fueron beta-hemolítico (18.63%), con prevalencia de 10.12%. Peces con Streptococcus beta-hemolíticos presentaron epicarditis, perisplenitis y meningitis crónica, panoftalmitis, necrosis coagulativa del músculo esquelético y formación de granulomas. En el análisis de PCR en tiempo real, no se obtuvo ninguna tilapia positiva para S. iniae. Los resultados se analizaron mediante una simulación estocástica de la distribución beta usando el programa de incertidumbre @Risk, reportando una prevalencia media de 0.66% en tilapias enfermas. Conclusiones. Los peces analizados fueron positivos para bacterias del género Streptococcus, lo que confirma su presencia en la piscifactoría. Sin embargo, 19 aislados fueron beta-hemolíticos, y la presencia de S. iniae no fue positiva para la prevalencia límite de 2.7% en PCR en tiempo real

Influence of glyceryl guaiacolate ether on anesthetics in tilapia compared to benzocaine and eugenol

ABSTRACT Objective. The study aimed to investigate the effectiveness of glyceryl guaiacolate ether (GGE) and compare the times of induction, recovery, hematological changes, total protein and glycaemia among anesthetics in Nile tilapia, Oreochromis niloticus. Materials and methods. A total of 60 tilapia distributed in 3 aquariums (N=20) were used, which formed the group benzocaine (100 mg/L), eugenol (50 mg/L) and guaiacol glyceryl ether (9.000 mg/L). After the induction of anesthesia fish blood samples were collected to determine the complete hemogram and glycemia. Then the animals were placed in aquariums with running water for assessing the anesthesia recovery. Results. It was verified that GGE showed longer induction and recovery times as well a significant increase (p0.05). An increase in the number of monocytes in the group treated with benzocaine (p <0.05) was observed in the analysis of the hematological parameters with no difference between groups for other variables. Conclusions. Eugenol and benzocaine allow rapid induction and recovery in Nile tilapia, without evidence of stress during handling and GGE showed high induction and recovery times, being inadequate for anesthetic use in Nile tilapia

Vaccine protection against simian immunodeficiency virus in monkeys using recombinant gamma-2 herpesvirus

Recombinant strains of replication-competent rhesus monkey rhadinovirus (RRV) were constructed in which strong promoter/enhancer elements were used to drive expression of simian immunodeficiency virus (SIV) Env or Gag or a Rev-Tat-Nef fusion protein. Cultured rhesus monkey fibroblasts infected with each recombinant strain were shown to express the expected protein. Three RRV-negative and two RRV-positive rhesus monkeys were inoculated intravenously with a mixture of these three recombinant RRVs. Expression of SIV Gag was readily detected in lymph node biopsy specimens taken at 3 weeks postimmunization. Impressive anti-SIV cellular immune responses were elicited on the basis of major histocompatibility complex (MHC) tetramer staining and gamma interferon enzyme-linked immunospot (ELISPOT) assays. Responses were much greater in magnitude in the monkeys that were initially RRV negative but were still readily detected in the two monkeys that were naturally infected with RRV at the time of immunization. By 3 weeks postimmunization, responses measured by MHC tetramer staining in the two Mamu-A*01(+) RRV-negative monkeys reached 9.3% and 13.1% of all CD8(+) T cells in peripheral blood to the Gag CM9 epitope and 2.3% and 7.3% of all CD8(+) T cells in peripheral blood to the Tat SL8 epitope. Virus-specific CD8(+) T cell responses persisted at high levels up to the time of challenge at 18 weeks postimmunization, and responding cells maintained an effector memory phenotype. Despite the ability of the RRVenv recombinant to express high levels of Env in cultured cells, and despite the appearance of strong anti-RRV antibody responses in immunized monkeys, anti-Env antibody responses were below our ability to detect them. Immunized monkeys, together with three unimmunized controls, were challenged intravenously with 10 monkey infectious doses of SIVmac239. All five immunized monkeys and all three controls became infected with SIV, but peak viral loads were 1.2 to 3.0 log(10) units lower and chronic-phase viral loads were 1.0 to 3.0 log(10) units lower in immunized animals than the geometric mean of unimmunized controls. These differences were statistically significant. Anti-Env antibody responses following challenge indicated an anamnestic response in the vaccinated monkeys. These findings further demonstrate the potential of recombinant herpesviruses as preventive vaccines for AIDS. We hypothesize that this live, replication-competent, persistent herpesvirus vector could match, or come close to matching, live attenuated strains of SIV in the degree of protection if the difficulty with elicitation of anti-Env antibody responses can be overcome

Computing the effective action with the functional renormalization group

The \u201cexact\u201d or \u201cfunctional\u201d renormalization group equation describes the renormalization group flow of the effective average action \u393 k. The ordinary effective action \u393 0 can be obtained by integrating the flow equation from an ultraviolet scale k= \u39b down to k= 0. We give several examples of such calculations at one-loop, both in renormalizable and in effective field theories. We reproduce the four-point scattering amplitude in the case of a real scalar field theory with quartic potential and in the case of the pion chiral Lagrangian. In the case of gauge theories, we reproduce the vacuum polarization of QED and of Yang\u2013Mills theory. We also compute the two-point functions for scalars and gravitons in the effective field theory of scalar fields minimally coupled to gravity. \ua9 2016, The Author(s)