RESUMEN
Although brain scars in adults have been extensively studied, there is less data available regarding scar formation during the neonatal period, and the involvement of peripheral immune cells in this process remains unexplored in neonates. Using a murine model of neonatal hypoxic-ischemic encephalopathy (HIE) and confocal microscopy, we characterized the scarring process and examined the recruitment of peripheral immune cells to cortical and hippocampal scars for up to 1 year post-insult. Regional differences in scar formation were observed, including the presence of reticular fibrotic networks in the cortex and perivascular fibrosis in the hippocampus. We identified chemokines with chronically elevated levels in both regions and demonstrated, through a parabiosis-based strategy, the recruitment of lymphocytes, neutrophils, and monocyte-derived macrophages to the scars several weeks after the neonatal insult. After 1 year, however, neutrophils and lymphocytes were absent from the scars. Our data indicate that peripheral immune cells are transient components of HIE-induced brain scars, opening up new possibilities for late therapeutic interventions.
Asunto(s)
Cicatriz , Hipoxia-Isquemia Encefálica , Adulto , Animales , Humanos , Ratones , Cicatriz/patología , Encéfalo/patología , Macrófagos , Hipoxia-Isquemia Encefálica/patologíaRESUMEN
The vast spectrum of clinical features of COVID-19 keeps challenging scientists and clinicians. Low resistance to infection might result in long-term viral persistence, but the underlying mechanisms remain unclear. Here, we studied the immune response of immunocompetent COVID-19 patients with prolonged SARS-CoV-2 infection by immunophenotyping, cytokine and serological analysis. Despite viral loads and symptoms comparable to regular mildly symptomatic patients, long-term carriers displayed weaker systemic IFN-I responses and fewer circulating pDCs and NK cells at disease onset. Type 1 cytokines remained low, while type-3 cytokines were in turn enhanced. Of interest, we observed no defects in antigen-specific cytotoxic T cell responses, and circulating antibodies displayed higher affinity against different variants of SARS-CoV-2 Spike protein in these patients. The identification of distinct immune responses in long-term carriers adds up to our understanding of essential host protective mechanisms to ensure tissue damage control despite prolonged viral infection.
RESUMEN
Despite the intramuscular route being the most used vaccination strategy against SARS-CoV-2, the intradermal route has been studied around the globe as a strong candidate for immunization against SARS-CoV-2. Adjuvants have shown to be essential vaccine components that are capable of driving robust immune responses and increasing the vaccination efficacy. In this work, our group aimed to develop a vaccination strategy for SARS-CoV-2 using a trimeric spike protein, by testing the best route with formulations containing the adjuvants AddaS03, CpG, MPL, Alum, or a combination of two of them. Our results showed that formulations that were made with AddaS03 or CpG alone or AddaS03 combined with CpG were able to induce high levels of IgG, IgG1, and IgG2a; high titers of neutralizing antibodies against SARS-CoV-2 original strain; and also induced high hypersensitivity during the challenge with Spike protein and a high level of IFN-γ producing CD4+ T-cells in mice. Altogether, those data indicate that AddaS03, CpG, or both combined may be used as adjuvants in vaccines for COVID-19.
RESUMEN
Serological tests detect antibodies generated by infection or vaccination, and are indispensable tools along different phases of a pandemic, from early monitoring of pathogen spread up to seroepidemiological studies supporting immunization policies. This work discusses the development of an accurate and affordable COVID-19 antibody test, from production of a recombinant protein antigen up to test validation and economic analysis. We first developed a cost-effective, scalable technology to produce SARS-COV-2 spike protein and then used this antigen to develop an enzyme-linked immunosorbent assay (ELISA). A receiver operator characteristic (ROC) analysis allowed optimizing the cut-off and confirmed the high accuracy of the test: 98.6% specificity and 95% sensitivity for 11+ days after symptoms onset. We further showed that dried blood spots collected by finger pricking on simple test strips could replace conventional plasma/serum samples. A cost estimate was performed and revealed a final retail price in the range of one US dollar, reflecting the low cost of the ELISA test platform and the elimination of the need for venous blood sampling and refrigerated sample handling in clinical laboratories. The presented workflow can be completed in 4 months from first antigen expression to final test validation. It can be applied to other pathogens and in future pandemics, facilitating reliable and affordable seroepidemiological surveillance also in remote areas and in low-income countries.
RESUMEN
The SARS-CoV-2 pandemic has had a social and economic impact worldwide, and vaccination is an efficient strategy for diminishing those damages. New adjuvant formulations are required for the high vaccine demands, especially adjuvant formulations that induce a Th1 phenotype. Herein we assess a vaccination strategy using a combination of Alum and polyinosinic:polycytidylic acid [Poly(I:C)] adjuvants plus the SARS-CoV-2 spike protein in a prefusion trimeric conformation by an intradermal (ID) route. We found high levels of IgG anti-spike antibodies in the serum by enzyme linked immunosorbent assay (ELISA) and high neutralizing titers against SARS-CoV-2 in vitro by neutralization assay, after two or three immunizations. By evaluating the production of IgG subtypes, as expected, we found that formulations containing Poly(I:C) induced IgG2a whereas Alum did not. The combination of these two adjuvants induced high levels of both IgG1 and IgG2a. In addition, cellular immune responses of CD4+ and CD8+ T cells producing interferon-gamma were equivalent, demonstrating that the Alum + Poly(I:C) combination supported a Th1 profile. Based on the high neutralizing titers, we evaluated B cells in the germinal centers, which are specific for receptor-binding domain (RBD) and spike, and observed that more positive B cells were induced upon the Alum + Poly(I:C) combination. Moreover, these B cells produced antibodies against both RBD and non-RBD sites. We also studied the impact of this vaccination preparation [spike protein with Alum + Poly(I:C)] in the lungs of mice challenged with inactivated SARS-CoV-2 virus. We found a production of IgG, but not IgA, and a reduction in neutrophil recruitment in the bronchoalveolar lavage fluid (BALF) of mice, suggesting that our immunization scheme reduced lung inflammation. Altogether, our data suggest that Alum and Poly(I:C) together is a possible adjuvant combination for vaccines against SARS-CoV-2 by the intradermal route.
Asunto(s)
COVID-19 , Vacunas Virales , Adyuvantes Inmunológicos , Compuestos de Alumbre , Animales , Linfocitos T CD8-positivos , Vacunas contra la COVID-19 , Humanos , Inmunoglobulina G , Ratones , Poli I-C , SARS-CoV-2 , Glicoproteína de la Espiga del CoronavirusRESUMEN
Besides antigen-specific responses to viral antigens, humoral immune response in virus infection can generate polyreactive and autoreactive antibodies. Dengue and Zika virus infections have been linked to antibody-mediated autoimmune disorders, including Guillain-Barré syndrome. A unique feature of flaviviruses is the secretion of nonstructural protein 1 (NS1) by infected cells. NS1 is highly immunogenic, and antibodies targeting NS1 can have both protective and pathogenic roles. In the present study, we investigated the humoral immune response to Zika virus NS1 and found NS1 to be an immunodominant viral antigen associated with the presence of autoreactive antibodies. Through single B cell cultures, we coupled binding assays and BCR sequencing, confirming the immunodominance of NS1. We demonstrate the presence of self-reactive clones in germinal centers after both infection and immunization, some of which present cross-reactivity with NS1. Sequence analysis of anti-NS1 B cell clones showed sequence features associated with pathogenic autoreactive antibodies. Our findings demonstrate NS1 immunodominance at the cellular level as well as a potential role for NS1 in ZIKV-associated autoimmune manifestations.
Asunto(s)
Reacciones Cruzadas/inmunología , Proteínas no Estructurales Virales/inmunología , Infección por el Virus Zika/inmunología , Animales , Anticuerpos Antivirales/inmunología , Antígenos Virales/inmunología , Linfocitos B/virología , Femenino , Centro Germinal/patología , Centro Germinal/virología , Inmunización , Inmunoglobulina M/sangre , Ratones Endogámicos BALB C , Proteínas no Estructurales Virales/sangre , Infección por el Virus Zika/virologíaRESUMEN
The dynamics underlying severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) reinfection remain poorly understood. We identified a small cluster of patients in Brazil who experienced 2 episodes of coronavirus disease (COVID-19) in March and late May 2020. In the first episode, patients manifested an enhanced innate response compared with healthy persons, but neutralizing humoral immunity was not fully achieved. The second episode was associated with different SARS-CoV-2 strains, higher viral loads, and clinical symptoms. Our finding that persons with mild COVID-19 may have controlled SARS-CoV-2 replication without developing detectable humoral immunity suggests that reinfection is more frequent than supposed, but this hypothesis is not well documented.
Asunto(s)
COVID-19 , SARS-CoV-2 , Brasil/epidemiología , Humanos , Inmunidad Humoral , ReinfecciónRESUMEN
Background: Convalescent plasma is a potential therapeutic option for critically ill patients with coronavirus disease 19 (COVID-19), yet its efficacy remains to be determined. The aim was to investigate the effects of convalescent plasma (CP) in critically ill patients with COVID-19. Methods: This was a single-center prospective observational study conducted in Rio de Janeiro, Brazil, from March 17th to May 30th, with final follow-up on June 30th. We included 113 laboratory-confirmed COVID-19 patients with respiratory failure. Primary outcomes were time to clinical improvement and survival within 28 days. Secondary outcomes included behavior of biomarkers and viral loads. Kaplan-Meier analyses and Cox proportional-hazards regression using propensity score with inverse-probability weighing were performed. Results: 41 patients received CP and 72 received standard of care (SOC). Median age was 61 years (IQR 48-68), disease duration was 10 days (IQR 6-13), and 86% were mechanically ventilated. At least 29 out of 41CP-recipients had baseline IgG titers ≥ 1:1,080. Clinical improvement within 28 days occurred in 19 (46%) CP-treated patients, as compared to 23 (32%) in the SOC group [adjusted hazard ratio (aHR) 0.91 (0.49-1.69)]. There was no significant change in 28-day mortality (CP 49% vs. SOC 56%; aHR 0.90 [0.52-1.57]). Biomarker assessment revealed reduced inflammatory activity and increased lymphocyte count after CP. Conclusions: In this study, CP was not associated with clinical improvement or increase in 28-day survival. However, our study may have been underpowered and included patients with high IgG titers and life-threatening disease. Clinical Trial Registration: The study protocol was retrospectively registered at the Brazilian Registry of Clinical Trials (ReBEC) with the identification RBR-4vm3yy (http://www.ensaiosclinicos.gov.br).
RESUMEN
Extracellular vesicles (EVs) are lipid bilayered compartments released by virtually all living cells, including fungi. Among the diverse molecules carried by fungal EVs, a number of immunogens, virulence factors and regulators have been characterised. Within EVs, these components could potentially impact disease outcomes by interacting with the host. From this perspective, we previously demonstrated that EVs from Candida albicans could be taken up by and activate macrophages and dendritic cells to produce cytokines and express costimulatory molecules. Moreover, pre-treatment of Galleria mellonella larvae with fungal EVs protected the insects against a subsequent lethal infection with C. albicans yeasts. These data indicate that C. albicans EVs are multi-antigenic compartments that activate the innate immune system and could be exploited as vaccine formulations. Here, we investigated whether immunisation with C. albicans EVs induces a protective effect against murine candidiasis in immunosuppressed mice. Total and fungal antigen-specific serum IgG antibodies increased by 21 days after immunisation, confirming the efficacy of the protocol. Vaccination decreased fungal burden in the liver, spleen and kidney of mice challenged with C. albicans. Splenic levels of cytokines indicated a lower inflammatory response in mice immunised with EVs when compared with EVs + Freund's adjuvant (ADJ). Higher levels of IL-12p70, TNFα and IFNγ were detected in mice vaccinated with EVs + ADJ, while IL-12p70, TGFß, IL-4 and IL-10 were increased when no adjuvants were added. Full protection of lethally challenged mice was observed when EVs were administered, regardless the presence of adjuvant. Physical properties of the EVs were also investigated and EVs produced by C. albicans were relatively stable after storage at 4, -20 or -80°C, keeping their ability to activate dendritic cells and to protect G. mellonella against a lethal candidiasis. Our data suggest that fungal EVs could be a safe source of antigens to be exploited in vaccine formulations.
Asunto(s)
Candida albicans/inmunología , Candidiasis/inmunología , Vesículas Extracelulares/inmunología , Animales , Anticuerpos Antifúngicos/sangre , Antígenos Fúngicos/inmunología , Candidiasis/prevención & control , Frío , Citocinas/sangre , Células Dendríticas/inmunología , Células Dendríticas/metabolismo , Femenino , Vacunas Fúngicas/inmunología , Inmunoglobulina G/sangre , Inmunoglobulina M/sangre , Interleucina-6/biosíntesis , Ratones , Ratones Endogámicos BALB C , Mariposas Nocturnas/inmunología , Mariposas Nocturnas/microbiología , VacunaciónRESUMEN
The relative potency and quality of mouse B cell response to Toll-like receptors (TLRs) signaling varies significantly depending on the B cell subset and on the TLR member being engaged. Although it has been shown that marginal zone cells respond faster than follicular (FO) splenic B cells to TLR4 stimulus, FO B cells retain full capacity to proliferate and generate plasmablasts and plasma cells (PBs/PCs) with 2-3 days delayed kinetics. It is not clear whether this scenario could be extended to other members of the TLR family. Here, using quantitative cell culture conditions optimized for B cell growth and differentiation, we show that TLR9 signaling by CpG, while promoting vigorous proliferation, completely fails to induce differentiation of FO B cells into PBs/PCs. Little or absent Ig secretion following TLR9 stimulus was accompanied by lack of expression of cell surface markers and canonical transcription factors involved in PB/PC differentiation. Moreover, not only TLR9 did not induce plasmocyte differentiation, but it also strongly inhibited the massive PB/PC differentiation of FO B cells triggered by LPS/TLR4. Our study reveals unexpected opposite roles for TLR4 and TLR9 in the control of plasma cell differentiation program and disagrees with previous conclusions obtained in high-density cultures conditions on the generation of plasmocytes by TRL9 signaling. The potential implications of these findings on the role of TLR9 in controlling self-tolerance, clonal sizes and regulation of humoral responses are discussed.
Asunto(s)
Linfocitos B/inmunología , Diferenciación Celular/inmunología , Células Plasmáticas/inmunología , Receptor Toll-Like 9/inmunología , Animales , Linfocitos B/metabolismo , Diferenciación Celular/genética , Proliferación Celular/efectos de los fármacos , Proliferación Celular/genética , Células Cultivadas , Ratones Endogámicos C57BL , Ratones Noqueados , Oligodesoxirribonucleótidos/inmunología , Oligodesoxirribonucleótidos/farmacología , Células Plasmáticas/metabolismo , Transducción de Señal/efectos de los fármacos , Transducción de Señal/genética , Transducción de Señal/inmunología , Receptor Toll-Like 9/agonistas , Receptor Toll-Like 9/genéticaRESUMEN
This work aims to study the immunomodulation of B lymphocytes during L. amazonensis infection. We demonstrated in this study that follicular B cells from draining lymph nodes of infected wild type BALB/c mice are the major source of IL-10 during infection. We infected BALB/Xid mice that developed smaller lesions in comparison with the control, but the parasite load obtained from the infected tissues was similar in both groups. We observed a reduction in the number of follicular B cells from BALB/Xid mice in relation to WT mice and, consequently, lower levels of IgM, IgG, IgG1, IgG2a and IgG2b in the serum of BALB/Xid when compared with wild type mice. BALB/Xid mice also presented lower levels of IL-10 in the infected footpad, draining lymph nodes and in the spleen when compared with WT infected tissues. We did not detect differences in the number of IL-10 producing CD4+ and CD8+ T cells between WT and BALB/Xid mice; however, a strong reduction of IL-10 producing follicular B cells was noted in BALB/Xid mice. When analyzed together, our data indicate that B cells are related with lesion pathogenesis through the production of antibodies and IL-10.
Asunto(s)
Linfocitos B/inmunología , Inmunomodulación/inmunología , Interleucina-10/inmunología , Leishmania mexicana/inmunología , Leishmaniasis Cutánea/inmunología , Animales , Linfocitos T CD4-Positivos/inmunología , Linfocitos T CD4-Positivos/parasitología , Linfocitos T CD8-positivos/inmunología , Linfocitos T CD8-positivos/parasitología , Inmunoglobulinas/inmunología , Leishmaniasis Cutánea/parasitología , Ganglios Linfáticos/inmunología , Ratones , Ratones Endogámicos BALB C , Piel/inmunología , Piel/parasitología , Bazo/inmunología , Bazo/parasitologíaRESUMEN
Protective adaptive immunity to Zika virus (ZIKV) has been mainly attributed to cytotoxic CD8+ T cells and neutralizing antibodies, while the participation of CD4+ T cells in resistance has remained largely uncharacterized. Here, we show a neutralizing antibody response, dependent on CD4+ T cells and IFNγ signaling, which we detected during the first week of infection and is associated with reduced viral load in the brain, prevention of rapid disease onset and survival. We demonstrate participation of these components in the resistance to ZIKV during primary infection and in murine adoptive transfer models of heterologous ZIKV infection in a background of IFNR deficiency. The protective effect of adoptively transferred CD4+ T cells requires IFNγ signaling, CD8+ T cells and B lymphocytes in recipient mice. Together, this indicates the importance of CD4+ T cell responses in future vaccine design for ZIKV.
Asunto(s)
Inmunidad Adaptativa , Traslado Adoptivo , Anticuerpos Antivirales/inmunología , Linfocitos T CD4-Positivos/inmunología , Interferón gamma/metabolismo , Infección por el Virus Zika/inmunología , Animales , Anticuerpos Neutralizantes/inmunología , Peso Corporal , Chlorocebus aethiops , Femenino , Inmunoglobulina G , Masculino , Ratones , Células Vero , Virus ZikaRESUMEN
Estudos acerca da morfologia de animais silvestres servem de subsídio para trabalhos de manejo e preservação de diferentes espécies, pois fornecem informações para a tomada de medidas que auxiliem na manutenção destes em cativeiro, na preservação em habitat natural ou mesmo para ações voltadas a reintrodução ao habitat de origem. Estudos referentes à morfologia de cutias abordam os diversos sistemas, mas nenhum faz referência à arquitetura ou estrutura de suas glândulas salivares. Assim este trabalho objetivou descrever macro e microscopicamente as glândulas salivares maiores de cutias. Foram utilizados dez animais adultos para o desenvolvimento de metodologias relativas à macroscopia propriamente dita das glândulas, microscopia de luz, microscopia eletrônica de transmissão e microscopia eletrônica de varredura. Foram identificadas quatro glândulas salivares maiores nos animais estudados, denominadas parótida, mandibular, zigomática e sublingual. As glândulas apresentaram-se como sendo do tipo tubuloacinares e contendo em seu parênquima ductos dos mais variados tamanhos. Com exceção da glândula parótida, que era estritamente serosa, as demais eram mistas. Da mesma forma, apenas a glândula mandibular foi identificada a presença de ducto do tipo granuloso. Apresentando as cutias os quatro pares de glândulas salivares maiores, estes animais podem servir de modelo para os estudos acerca das mudanças anatômicas sofridas pelos roedores para se adaptar aos diversos habitat do planeta.(AU)
Studies on wild animal morphology serve as theoretical basis for the management and conservation of different species, because they provide necessary information for measures to keep these animals in captivity, in their natural habitat or even to reintroduce them into their original habitat. Studies about the morphology of the red-rumped agouti, Dasyprocta leporina, approach the various organic systems, but not a single study refers to topography and structure arrangement of their salivary glands. Thus, this paper aimed to gross and microscopic description of the larger salivary glands of red-rumped agouti. Ten adult D. leporina were used to study the macroscopic aspect of the glands, as well as the microscopic aspects with light microscopy, scanning and transmission electronic microscopy. Four larger salivary glands were identified: parotid glands, mandibular glands, zygomatic glands and sublingual glands. The tubuloacinar glands contained in their parenchyma ducts of extremely varied sizes. With exception of the strictly serous parotid glands, the others were mixed, and only the mandibular glands had granulous ducts. The red-rumped agouti with four pairs of larger salivary glands may be a model for studies concerning the anatomical changes in rodents for adaptation to various habitats.(AU)
Asunto(s)
Animales , Dasyproctidae , Glándulas Salivales , Roedores , Animales Salvajes , Glándulas Salivales/anatomía & histología , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Microscopía/veterinariaRESUMEN
Estudos acerca da morfologia de animais silvestres servem de subsídio para trabalhos de manejo e preservação de diferentes espécies, pois fornecem informações para a tomada de medidas que auxiliem na manutenção destes em cativeiro, na preservação em habitat natural ou mesmo para ações voltadas a reintrodução ao habitat de origem. Estudos referentes à morfologia de cutias abordam os diversos sistemas, mas nenhum faz referência à arquitetura ou estrutura de suas glândulas salivares. Assim este trabalho objetivou descrever macro e microscopicamente as glândulas salivares maiores de cutias. Foram utilizados dez animais adultos para o desenvolvimento de metodologias relativas à macroscopia propriamente dita das glândulas, microscopia de luz, microscopia eletrônica de transmissão e microscopia eletrônica de varredura. Foram identificadas quatro glândulas salivares maiores nos animais estudados, denominadas parótida, mandibular, zigomática e sublingual. As glândulas apresentaram-se como sendo do tipo tubuloacinares e contendo em seu parênquima ductos dos mais variados tamanhos. Com exceção da glândula parótida, que era estritamente serosa, as demais eram mistas. Da mesma forma, apenas a glândula mandibular foi identificada a presença de ducto do tipo granuloso. Apresentando as cutias os quatro pares de glândulas salivares maiores, estes animais podem servir de modelo para os estudos acerca das mudanças anatômicas sofridas pelos roedores para se adaptar aos diversos habitat do planeta.
Studies on wild animal morphology serve as theoretical basis for the management and conservation of different species, because they provide necessary information for measures to keep these animals in captivity, in their natural habitat or even to reintroduce them into their original habitat. Studies about the morphology of the red-rumped agouti, Dasyprocta leporina, approach the various organic systems, but not a single study refers to topography and structure arrangement of their salivary glands. Thus, this paper aimed to gross and microscopic description of the larger salivary glands of red-rumped agouti. Ten adult D. leporina were used to study the macroscopic aspect of the glands, as well as the microscopic aspects with light microscopy, scanning and transmission electronic microscopy. Four larger salivary glands were identified: parotid glands, mandibular glands, zygomatic glands and sublingual glands. The tubuloacinar glands contained in their parenchyma ducts of extremely varied sizes. With exception of the strictly serous parotid glands, the others were mixed, and only the mandibular glands had granulous ducts. The red-rumped agouti with four pairs of larger salivary glands may be a model for studies concerning the anatomical changes in rodents for adaptation to various habitats.
Asunto(s)
Animales , Dasyproctidae , Roedores , Glándulas Salivales , Animales Salvajes , Glándulas Salivales/anatomía & histología , Microscopía Electrónica de Rastreo , Microscopía Electrónica de Transmisión , Microscopía/veterinariaRESUMEN
O objetivo deste trabalho foi estudar o período de inversão do saco vitelino bem como a dinâmica resultante deste processo na gestação inicial em preás, utilizando-se microscopia de luz, microscopia eletrônica de varredura e de transmissão. No décimo segundo dia de gestação observou-se o desenvolvimento dos endodermas parietal e visceral delimitando a cavidade do saco vitelino. O endoderma parietal foi evidenciado revestindo a superfície fetal da placenta corioalantoidea bem como contornando o espaço delimitado pela decídua capsular. Estes endodermas apresentaram formato prismático e encontraram-se separados do trofoblasto por uma desenvolvida membrana de Reichert. Já o endoderma visceral continha vasos vitelínicos e possuía vilosidades apenas em determinadas áreas. No décimo quarto dia de gestação verificou-se a inversão do saco vitelino, caracterizada pela degeneração do endoderma parietal e trofoblasto mural, associado ao desaparecimento gradual da membrana de Reichert. Como consequência deste fenômeno, o endoderma visceral passou a constituir uma interface com o epitélio uterino. Após a inversão, o endoderma parietal que permaneceu íntegro foi aquele que se apoiava na superfície da placenta corioalantóidea, apresentando células em formato colunar alto e característica de epitélio pseudoestratificado. O endoderma visceral apresentou numerosas vilosidades apicais principalmente em regiões próximas a placenta corioalantóidea. Com o contínuo desenvolvimento do embrião e placenta corioalantóidea, observou-se o surgimento de importante área de aposição entre os endodermas visceral e parietal. A inversão do saco vitelino representou uma disposição anatômica favorável ao desenvolvimento embrionário, além de ser uma característica evolutiva nesta espécie de roedor.
The aim of this study was to study the time of yolk sac inversion as well as the dynamics resulting from this process in galea throughout pregnancy. For this, conventional histological techniques, scanning electron microscopy and transmission electron microscopy were used. Parietal and visceral endoderm delimiting the yolk sac cavity was observed at 12 days of pregnancy. The parietal endoderm was coating the fetal surface of the chorioallantoic placenta as well as delimiting the decidua capsularis area. This endoderm had prismatic format and were apart from the trophoblast by an enlarged Reichert's membrane. The visceral endoderm had vitelline vessels and there were villi only in certain areas. At 14 days of pregnancy the yolk sac inversion was characterized by the degeneration of parietal endoderm and mural trophoblast, and also the gradual disappearance of the Reichert's membrane. So it made the visceral endoderm establish an interface with the uterine epithelium. After the inversion, the parietal endoderm which remained intact was the one that rested on the chorioallantoic placenta surface. It presented cells with high columnar format and pseudostratified epithelium featured. The visceral endoderm presented many apical villi, especially in areas close to the chorioallantoic placenta. The continued development of the embryo and chorioallantoic placenta evidenced the emergence of an important apposition area between visceral and parietal endoderm. The yolk sac inversion represented an anatomical arrangement in favor of the embryo development as well as an evolutionary trait in this rodent species.
Asunto(s)
Animales , Endodermo/embriología , Saco Vitelino/anatomía & histología , Cobayas/clasificación , Embrión de Mamíferos/embriologíaRESUMEN
O objetivo deste trabalho foi estudar o período de inversão do saco vitelino bem como a dinâmica resultante deste processo na gestação inicial em preás, utilizando-se microscopia de luz, microscopia eletrônica de varredura e de transmissão. No décimo segundo dia de gestação observou-se o desenvolvimento dos endodermas parietal e visceral delimitando a cavidade do saco vitelino. O endoderma parietal foi evidenciado revestindo a superfície fetal da placenta corioalantoidea bem como contornando o espaço delimitado pela decídua capsular. Estes endodermas apresentaram formato prismático e encontraram-se separados do trofoblasto por uma desenvolvida membrana de Reichert. Já o endoderma visceral continha vasos vitelínicos e possuía vilosidades apenas em determinadas áreas. No décimo quarto dia de gestação verificou-se a inversão do saco vitelino, caracterizada pela degeneração do endoderma parietal e trofoblasto mural, associado ao desaparecimento gradual da membrana de Reichert. Como consequência deste fenômeno, o endoderma visceral passou a constituir uma interface com o epitélio uterino. Após a inversão, o endoderma parietal que permaneceu íntegro foi aquele que se apoiava na superfície da placenta corioalantóidea, apresentando células em formato colunar alto e característica de epitélio pseudoestratificado. O endoderma visceral apresentou numerosas vilosidades apicais principalmente em regiões próximas a placenta corioalantóidea. Com o contínuo desenvolvimento do embrião e placenta corioalantóidea, observou-se o surgimento de importante área de aposição entre os endodermas visceral e parietal. A inversão do saco vitelino representou uma disposição anatômica favorável ao desenvolvimento embrionário, além de ser uma característica evolutiva nesta espécie de roedor.(AU)
The aim of this study was to study the time of yolk sac inversion as well as the dynamics resulting from this process in galea throughout pregnancy. For this, conventional histological techniques, scanning electron microscopy and transmission electron microscopy were used. Parietal and visceral endoderm delimiting the yolk sac cavity was observed at 12 days of pregnancy. The parietal endoderm was coating the fetal surface of the chorioallantoic placenta as well as delimiting the decidua capsularis area. This endoderm had prismatic format and were apart from the trophoblast by an enlarged Reichert's membrane. The visceral endoderm had vitelline vessels and there were villi only in certain areas. At 14 days of pregnancy the yolk sac inversion was characterized by the degeneration of parietal endoderm and mural trophoblast, and also the gradual disappearance of the Reichert's membrane. So it made the visceral endoderm establish an interface with the uterine epithelium. After the inversion, the parietal endoderm which remained intact was the one that rested on the chorioallantoic placenta surface. It presented cells with high columnar format and pseudostratified epithelium featured. The visceral endoderm presented many apical villi, especially in areas close to the chorioallantoic placenta. The continued development of the embryo and chorioallantoic placenta evidenced the emergence of an important apposition area between visceral and parietal endoderm. The yolk sac inversion represented an anatomical arrangement in favor of the embryo development as well as an evolutionary trait in this rodent species.(AU)
Asunto(s)
Animales , Saco Vitelino/anatomía & histología , Endodermo/embriología , Cobayas/clasificación , Embrión de Mamíferos/embriologíaRESUMEN
Objetivou-se com este trabalho estudar o perfil hematológico de caprinos (Capra hircus) da raça Canindé, criados no Estado do Rio Grande do Norte, como também a busca de valores de referência que se adequem à nossa região. Foram coletadas amostras sanguíneas de 58 animais clinicamente sadios, distribuídos em 4 grupos (machos acima de 5 meses, fêmeas gestantes, fêmeas não gestantes e filhotes até 4 meses de idade). A partir de única amostra de sangue de cada animal foram realizados o eritrograma (contagem de hemácias, hematócrito, hemoglobina, volume corpuscular médio e concentração de hemoglobina corpuscular média) e o leucograma (contagem total e diferencial de leucócitos). Os dados obtidos foram avaliados por meio do teste de Tukey para variáveis paramétricas e Kruskal-Wallis seguido pelo teste de Dunn para não paramétricos em nível de significância de (p<0.05). Os resultados apontam para existência de maiores quantidades de He nos machos adultos e fêmeas não gestantes, diferente do que ocorre com o VCM; já com relação ao leucograma temos que os eosinófilos mostram-se mais elevados nas fêmeas adultas e os monócitos se elevam quando na presença da gestação. Os valores deste trabalho podem servir de referência para raça Canindé, tornando possível futuras interpretações para os parâmetros verificados, além de subsidiar novos estudos em animais hígidos ou doentes. Mostra a necessidade de pesquisas que evidenciem as condições semiáridas de manejo e alimentação, bem como a avaliação dos fatores de variação, sobre constituintes do sangue, que levem em conta o sexo, a idade e estado fisiológico dos animais.(AU)
The objective of this paper was to study the hematological profile of Canindé breed goats (Capra hircus) raised in the state of Rio Grande do Norte, as well as the acquirement of reference values appropriate to our region. Blood samples were collected from 58 clinically healthy animals, distributed in 4 groups (males, over 5-month-old, pregnant and not pregnant females and young goats, up to 4 months of age). From a single blood sample from each animal, it was possible to perform the red blood cell (RBC) count, packed cell volume (PCV), hemoglobin, mean cell volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and the white blood cell (WBC) count and differential). The obtained data was statically evaluated by the Tukey test for parametric variables and Kruskal-Wallis test followed by Dunn's test for nonparametric-level significance (p<0.05). The results of this study can be used as a reference for this breed of goats, making future interpretations possible from the evaluated parameters, and can support further studies in healthy or diseased animals. It shows the need for further research that demonstrates the semiarid conditions of handling and feeding, as well as assessment of variation factors on the constituents of blood taking into account sex, age and the physiological state of animals.(AU)
Asunto(s)
Animales , Cabras/sangre , Recuento de Células Sanguíneas/veterinaria , Zona Semiárida , Hematócrito/veterinaria , Leucocitos , Valores de ReferenciaRESUMEN
Objetivou-se com este trabalho estudar o perfil hematológico de caprinos (Capra hircus) da raça Canindé, criados no Estado do Rio Grande do Norte, como também a busca de valores de referência que se adequem à nossa região. Foram coletadas amostras sanguíneas de 58 animais clinicamente sadios, distribuídos em 4 grupos (machos acima de 5 meses, fêmeas gestantes, fêmeas não gestantes e filhotes até 4 meses de idade). A partir de única amostra de sangue de cada animal foram realizados o eritrograma (contagem de hemácias, hematócrito, hemoglobina, volume corpuscular médio e concentração de hemoglobina corpuscular média) e o leucograma (contagem total e diferencial de leucócitos). Os dados obtidos foram avaliados por meio do teste de Tukey para variáveis paramétricas e Kruskal-Wallis seguido pelo teste de Dunn para não paramétricos em nível de significância de (p<0.05). Os resultados apontam para existência de maiores quantidades de He nos machos adultos e fêmeas não gestantes, diferente do que ocorre com o VCM; já com relação ao leucograma temos que os eosinófilos mostram-se mais elevados nas fêmeas adultas e os monócitos se elevam quando na presença da gestação. Os valores deste trabalho podem servir de referência para raça Canindé, tornando possível futuras interpretações para os parâmetros verificados, além de subsidiar novos estudos em animais hígidos ou doentes. Mostra a necessidade de pesquisas que evidenciem as condições semiáridas de manejo e alimentação, bem como a avaliação dos fatores de variação, sobre constituintes do sangue, que levem em conta o sexo, a idade e estado fisiológico dos animais.(AU)
The objective of this paper was to study the hematological profile of Canindé breed goats (Capra hircus) raised in the state of Rio Grande do Norte, as well as the acquirement of reference values appropriate to our region. Blood samples were collected from 58 clinically healthy animals, distributed in 4 groups (males, over 5-month-old, pregnant and not pregnant females and young goats, up to 4 months of age). From a single blood sample from each animal, it was possible to perform the red blood cell (RBC) count, packed cell volume (PCV), hemoglobin, mean cell volume (MCV), mean corpuscular hemoglobin concentration (MCHC) and the white blood cell (WBC) count and differential). The obtained data was statically evaluated by the Tukey test for parametric variables and Kruskal-Wallis test followed by Dunn's test for nonparametric-level significance (p<0.05). The results of this study can be used as a reference for this breed of goats, making future interpretations possible from the evaluated parameters, and can support further studies in healthy or diseased animals. It shows the need for further research that demonstrates the semiarid conditions of handling and feeding, as well as assessment of variation factors on the constituents of blood taking into account sex, age and the physiological state of animals.(AU)
Asunto(s)
Animales , Rumiantes , Hemoglobinas , Cabras/sangre , Fármacos Hematológicos/análisisRESUMEN
Apesar de sua conhecida resistência, os ovinos estão sujeitos as endoparasitoses, sendo este o principal fator limitante para a sua produção em todo o mundo, especialmente em regiões tropicais. Este trabalho objetiva avaliar a carga parasitária, o perfil hematológico e bioquímico sérico, e o grau Famacha de ovinos sem padrão racial definido infectados e tratados com ivermectina e closantel. O trabalho foi realizado no mês de julho de 2010, numa propriedade rural no município de Mossoró-RN. Foram selecionados 41 animais, os quais foram divididos em três grupos: grupo I- controle (sem tratamento), grupo II- tratado com ivermectina (0,2mg/kg) e Grupo III- tratado com closantel (5mg/kg). As amostras de fezes foram coletadas nos dias 0, 7, 14 e 21, pós-tratamento para análise quantita tiva (OPG) e para análise qualitativa (coprocultura), foram utilizadas amostras dos dias 0, 14 e 21. A primeira coleta de sangue se deu no dia 0, a segunda e a terceira 24 e 72 horas após a primeira, respectivamente, a quarta e a quinta 7 e 14 dias após a primeira, dos quais foram analisados o hemograma, os níveis séricos de proteínas totais, albumina e globulinas, e a relação albumina/globulinas. Foi determinado o grau Famacha de cada animal em todos os momentos de coleta de amostras. O tratamento com closantel foi eficaz no tratamento das helmintoses, principalmente Haemonchus contortus, nos ovinos avaliados. Por outro lado, houve resistência dos parasitas à ivermectina. Os dados de Famacha apresentaram correlação negativa com hematócrito, leucócitos, hemoglobina, albumina, proteínas totais, globulinas e relação albumina/globulinas. O hematócrito apresentou uma correlação forte e positiva com Hemoglobina, Albumina e Proteínas totais. Os tratamentos com closantel e ivermectina não foram responsáveis por alterações consideráveis nos parâmetros hematológicos e bioquímicos avaliados.
Despite its known resistance, the sheep are subject to endoparasitoses, which are the main limiting factor for its production worldwide, especially in tropical regions. This study aims to evaluate the changes in parasitic load, serum biochemical and hematological panel, and Famacha scores of mixed-bred sheep naturally infected with nematodes and treated with ivermectin and closantel. The study was conducted at July 2010, in a farm in the municipality of Mossoró, Rio Grande do Norte state, Brazil. We selected 41 animals were divided into three groups: group I- control (without treatment), group II- treated with ivermectin (0.2 mg/kg), and Group III- treated with closantel (5mg/kg). Stool samples were collected on days 0, 7, 14 and 21 after treatment for quantitative analysis (EPG) and qualitative analysis (stool culture), we used samples from days 0, 14 and 21. The first blood sample was given on day 0, the second and third 24 and 72 hours after the first, respectively the fourth and fifth 7 and 14 days after the first of which analyzed the blood count, serum total protein, albumin and globulins, and albumin/globulins ratio. Famacha scores were determined the degree of each animal at all times of sampling. Closantel administration was effective in the treatment of helminthiasis, especially Haemonchus contortus in evaluated sheep. On the other hand, there was parasite resistance to ivermectin. Famacha data showed negative correlation with packed cell volume, leukocytes, hemoglobin, albumin, total protein, globulin and albumin/globulin ratio. The packed cell volume showed a strong and positive correlation with hemoglobin, albumin and total protein. Treatment with ivermectin and closantel were not responsible for considerable changes in hematological and biochemical parameters evaluated.
Asunto(s)
Animales , Antiparasitarios/uso terapéutico , Nematodos , Ovinos/parasitología , Técnicas y Procedimientos Diagnósticos/veterinaria , Recuento de Células Sanguíneas/veterinaria , Pruebas SerológicasRESUMEN
Apesar de sua conhecida resistência, os ovinos estão sujeitos as endoparasitoses, sendo este o principal fator limitante para a sua produção em todo o mundo, especialmente em regiões tropicais. Este trabalho objetiva avaliar a carga parasitária, o perfil hematológico e bioquímico sérico, e o grau Famacha de ovinos sem padrão racial definido infectados e tratados com ivermectina e closantel. O trabalho foi realizado no mês de julho de 2010, numa propriedade rural no município de Mossoró-RN. Foram selecionados 41 animais, os quais foram divididos em três grupos: grupo I- controle (sem tratamento), grupo II- tratado com ivermectina (0,2mg/kg) e Grupo III- tratado com closantel (5mg/kg). As amostras de fezes foram coletadas nos dias 0, 7, 14 e 21, pós-tratamento para análise quantita tiva (OPG) e para análise qualitativa (coprocultura), foram utilizadas amostras dos dias 0, 14 e 21. A primeira coleta de sangue se deu no dia 0, a segunda e a terceira 24 e 72 horas após a primeira, respectivamente, a quarta e a quinta 7 e 14 dias após a primeira, dos quais foram analisados o hemograma, os níveis séricos de proteínas totais, albumina e globulinas, e a relação albumina/globulinas. Foi determinado o grau Famacha de cada animal em todos os momentos de coleta de amostras. O tratamento com closantel foi eficaz no tratamento das helmintoses, principalmente Haemonchus contortus, nos ovinos avaliados. Por outro lado, houve resistência dos parasitas à ivermectina. Os dados de Famacha apresentaram correlação negativa com hematócrito, leucócitos, hemoglobina, albumina, proteínas totais, globulinas e relação albumina/globulinas. O hematócrito apresentou uma correlação forte e positiva com Hemoglobina, Albumina e Proteínas totais. Os tratamentos com closantel e ivermectina não foram responsáveis por alterações consideráveis nos parâmetros hematológicos e bioquímicos avaliados.(AU)
Despite its known resistance, the sheep are subject to endoparasitoses, which are the main limiting factor for its production worldwide, especially in tropical regions. This study aims to evaluate the changes in parasitic load, serum biochemical and hematological panel, and Famacha scores of mixed-bred sheep naturally infected with nematodes and treated with ivermectin and closantel. The study was conducted at July 2010, in a farm in the municipality of Mossoró, Rio Grande do Norte state, Brazil. We selected 41 animals were divided into three groups: group I- control (without treatment), group II- treated with ivermectin (0.2 mg/kg), and Group III- treated with closantel (5mg/kg). Stool samples were collected on days 0, 7, 14 and 21 after treatment for quantitative analysis (EPG) and qualitative analysis (stool culture), we used samples from days 0, 14 and 21. The first blood sample was given on day 0, the second and third 24 and 72 hours after the first, respectively the fourth and fifth 7 and 14 days after the first of which analyzed the blood count, serum total protein, albumin and globulins, and albumin/globulins ratio. Famacha scores were determined the degree of each animal at all times of sampling. Closantel administration was effective in the treatment of helminthiasis, especially Haemonchus contortus in evaluated sheep. On the other hand, there was parasite resistance to ivermectin. Famacha data showed negative correlation with packed cell volume, leukocytes, hemoglobin, albumin, total protein, globulin and albumin/globulin ratio. The packed cell volume showed a strong and positive correlation with hemoglobin, albumin and total protein. Treatment with ivermectin and closantel were not responsible for considerable changes in hematological and biochemical parameters evaluated.(AU)