Abstract A lectin from the marine sponge Haliclona (Reniera) implexiformis (HiL) was isolated by affinity chromatography on Sepharose™ matrix. HiL showed specificity for galactose and its derivatives. The glycoproteins porcine stomach mucin (PSM) and bovine stomach mucin (BSM) were potent inhibitors. Hemagglutinating activity of the lectin was maximal between pH 5.0 and 9.0. The lectin remained active until 60°C. The presence of CaCl2 and EDTA did not affect the hemagglutinating activity. In SDS-PAGE, HiL showed a single band of 20 kDa under reduced conditions, whereas in the non-reducing conditions, it showed a band of 20 kDa and one additional band of 36 kDa. The average molecular mass determined by Electrospray Ionization Mass Spectrometry (ESI-MS) was 35.874 ± 2 Da in native and non-reducing conditions, whereas carboxyamidomethylated-lectin showed 18,111 Da. These data indicated that HiL consists in a dimer formed by identical subunits linked by disulfide bonds. Partial amino acid sequence of HiL was determined by mass spectrometry, and revealed that it is a new type of lectin, which showed no similarity with any protein. Secondary structure consisted of 6% α-helice, 31% β-sheet, 18% β-turn and 45% random coil. HiL showed significant reduction in the number of viable cells of Staphylococcus biofilms. Reniera (Reniera (HiL Sepharose matrix derivatives PSM (PSM BSM (BSM inhibitors 50 5 0 5. 90 9 9.0 60C C 60 60°C CaCl SDSPAGE, SDSPAGE SDS PAGE, PAGE SDS-PAGE conditions nonreducing non reducing 3 ESIMS ESI MS (ESI-MS 35874 35 874 35.87 carboxyamidomethylatedlectin carboxyamidomethylated 18111 18 111 18,11 bonds spectrometry protein 6 αhelice, αhelice α helice, helice α-helice 31 βsheet, βsheet β sheet, sheet β-sheet βturn turn 45 coil biofilms 9. 3587 87 35.8 1811 1 11 18,1 4 358 8 35. 181 18,

Abstract Health professionals working to mitigate the COVID-19 pandemic are one of the main risk groups for the disease, being prioritized for vaccination. Considering this, the aim of this study was to analyze the immune response of these professionals immunized with CoronaVac in the first and second doses. Blood samples were collected after the first and second doses of the vaccine (CoronaVac) and used to investigate hematological and biochemical parameters, analysis of immunoglobulin production, cytokines, and gene expression profile, as well as the identification of subsets of immune cells. Post-first dose immunological phenotypic memory (CD27+) profiles (T CD4+, TCD8+ and CD19+) showed a significant increase, as did Monocyte APCs (CD80+HLA-DR+) in relation to the second dose. The cytokines IL-2, IL-6 and IFN-° showed increased values in relation to the other analyzed cytokines. The Th2/Th17 profile in the second dose was characterized by gene expression analysis. The production of IgM and IgG after vaccination showed statistically significant values in the comparison between doses. CoronaVac showed activation of APCs monocytes, memory response of T and B lymphocytes, with immunoglobulins production. This set of responses is characterized by the Th2/Th17 immunological profile.

RESUMO Mycobacterium avium subesp. paratuberculosis (MAP) e o vírus da leucemia bovina (BLV) são agentes que causam grandes perdas econômicas nos rebanhos. O objetivo deste estudo foi avaliar a situação epidemiológica da paratuberculose bovina (PTB) e leucose enzoótica bovina (EBL) em rebanhos leiteiros de Lagoa Formosa, Minas Gerais, Brasil. Foram coletadas 236 amostras de sangue de vacas, as quais foram submetidas aos testes ELISA e imunodifusão em gel de ágar para detecção de anticorpos contra MAP e BLV. A soroprevalência de anticorpos contra MAP e BVL foi de 20% para os rebanhos e 6% para os animais e de 85% para os rebanhos e 50,42% para os animais, respectivamente. A presença dessas enfermidades deve servir como um alerta para os produtores e veterinários, para que concentrem maior atenção na implementação de medidas higiênico-sanitárias, incorporando elementos de vigilância com base nos riscos identificados no estudo.

The transport of myo-inositol is the main mechanism for the maintenance of its high intracellular levels. We aimed to measure the mRNA and protein levels of myo-inositol cotransporters in the sciatic nerve (SN) and dorsal root ganglia (DRG) during experimental diabetes. Streptozotocin-induced (STZ; 4, 8, and 12 weeks; 65 mg/kg; ip) diabetic rats (DB) and age-matched euglycemic (E) rats were used for the analysis of mRNA and protein levels of sodium myo-inositol cotransporters 1, 2 (SMIT1, SMIT2) or H+/myo-inositol cotransporter (HMIT). There was a significant reduction in the mRNA levels for SMIT1 in the SN and DRG (by 36.9 and 31.0%) in the 4-week DB (DB4) group compared to the E group. SMIT2 was not expressed in SN. The mRNA level for SMIT2 was up-regulated only in the DRG in the DB4 group. On the other hand, the protein level of SMIT1 decreased by 42.5, 41.3, and 44.8% in the SN after 4, 8, and 12 weeks of diabetes, respectively. In addition, there was a decrease of 64.3 and 58.0% of HMIT in membrane and cytosolic fractions, respectively, in the SN of the DB4 group. In the DRG, there was an increase of 230 and 86.3% for SMIT1 and HMIT, respectively, in the DB12 group. The levels of the main inositol transporters, SMIT1 and HMIT, were greatly reduced in the SN but not in the DRG. SMIT-1 was selectively reduced in the sciatic nerve during experimental STZ-induced diabetes.

RESUMO: Objetivou-se no estudo identificar a ocorrência de infecção pelo vírus da artrite encefalite caprina (CAEV) em propriedades produtoras de leite caprino com sistema intensivo no estado de Minas Gerais. Foram avaliadas cinco propriedades, localizadas em cidades distintas, totalizando 1072 animais, sendo 48 machos e 1024 fêmeas de diferentes faixas etárias, das raças Toggenburg, Alpina e Saanen. O método de diagnóstico utilizado foi o de imunodifusão em ágar gel (IDGA), para detecção de anticorpos anti-CAEV, por ser o diagnóstico preconizado pela Organização Mundial de Saúde Animal (OIE). A ocorrência de anticorpos anti-CAEV nas propriedades estudadas foi de 49,5% (531/1072), entretanto a mesma variou de acordo com a propriedade. Foram observados os seguintes resultados por propriedade: propriedade 1 = 69,6% (156/224); propriedade 2 = 41,5% (47/113); propriedade 3 = 40,3% (63/156); propriedade 4 = 24,6% (18/73) e propriedade 5 = 48,8% (247/506). Do total de machos avaliados, 14,5% (7/48) apresentaram sorologia positiva. De acordo com os resultados, uma alta ocorrência de animais soropositivos foi identificada no estado de Minas Gerais, o qual possui um dos maiores rebanhos de cabras leiteiras do Brasil. Portanto, salienta-se a necessidade de se adotar medidas estratégias sanitárias para o controle da CAE, como a realização de exames rotineiros nas propriedades e a separação de animais infectados dos sadios. A exclusão de reprodutores positivos nas propriedades também é uma medida de controle, pois já foi demonstrado que estes são fontes de infecção importantes.

ABSTRACT: The objective of the present study was to identify the occurrence of caprine arthritis encephalitis virus (CAEV) infection in dairy goat farms with intensive system in Minas Gerais State. Five properties were evaluated, totaling 1072 animals, being 48 males and 1024 females of different ages and breeds (Toggenburg, Saanen and Alpine). The diagnostic method used was the agar gel immunodiffusion (AGID) test, to detect antibodies anti-CAEV, which is the diagnostic test recommended by World Organization for Animal Health - OIE. The occurrence of anti-CAEV antibodies in the properties was 49.5% (531/1072), however varied according to the farm. The anti-CAEV antibodies detection varied according to the property: Farm 1 = 69.6% (156/224); farm 2 = 41.5% (47/113); farm 3 = 40.3% (63/156); farm 4 = 24.6% (18/73) and farm 5 = 48.8% (247/506). Of the total sampled males, 14.5% (7/48) were serologically positive. According to the results, there is a high occurrence rate of CAEV-seropositive animals in Minas Gerais, which has one of the largest Brazilian dairy goat herds. Therefore, it is essential to adopt strategies for CAE control, such as a routine of diagnostics tests in the properties and the separation of healthy from infected goats. The elimination of positive breeding goats in the properties is also a measure of control, because it has been shown that these are an important route of CAEV transmission in dairy farms.

ABSTRACT The antimicrobial potential of extracts of bark and leaves of Cassia bakeriana Craib, Fabaceae, against aerobic and anaerobic oral bacteria was evaluated by the microdilution broth method. For crude ethanol extracts and organic fractions tested, the bark dichloromethane phase showed a significant antibacterial effect, with MIC values ranging from 12.5 to 100 µg/ml for most of the microorganisms tested. Thus, a bioassay-guided fractionation of this fraction was performed. This fractionation led to isolation of the 1,8-dihydroxy-anthraquinone-3-carboxylic acid, also known as cassic acid or rhein. It is the first time that this bioactive anthraquinone has been isolated from this plant. Rhein exhibited good selectivity and high activity against anaerobic microorganisms, with MIC values ranging between 3.12 µg/ml (11.0 µM) and 25 µg/ml (88.0 µM). These results were considered very promising since the most active samples and rhein showed greater selectivity against oral microorganisms than toxicity to Vero cells.

Bauhinia forficata is native to South America and used with relative success in the folk medicine in Brazil. The diversity, antibacterial activity, and extracellular hydrolytic enzymes of endophytic fungi associated with this plant were studied. Plant samples, which included leaves, sepals, stems, and seeds, were used. Ninety-five endophytic fungal were isolated (18 from leaves, 22 from sepals, 46 from stems, and nine from seeds), comprising 28 species. The most frequently isolated species were Acremonium curvulum (9.5%), Aspergillus ochraceus (7.37%), Gibberella fujikuroi (10.53%), Myrothecium verrucaria (10.53%) and Trichoderma piluliferum(7.37%). Diversity and species richness were higher in stem tissues, and Sorensen’s index of similarity between the tissues was low. Eleven fungi showed antibacterial activity. Aspergillus ochraceus, Gibberella baccata, Penicillium commune, and P. glabrum were those with the greatest antibacterial activity against Staphylococcus aureus and/or Streptococcus pyogenes. Thirteen species showed proteolytic activity, particularly Phoma putaminum. Fourteen species were cellulase positive, particularly the Penicillium species and Myrmecridium schulzeri. All isolates tested were xylanase positive and 10 showed lipolytic activity, especially Penicillium glabrum. It is clear that the endophytic fungi from B. forficata have potential for the production of bioactive compounds and may be a source of new therapeutic agents for the effective treatment of diseases in humans, other animals, and plants. To our knowledge, this is the first study of endophytic fungi from different tissues of B. forficata and their biotechnological potential.

O presente artigo objetivou estudar a viabilidade da técnica de plasmaférese automatizada e padronizá-la em cinco equinos hígidos, bem como apresentar as complicações durante o procedimento, as adequações em relação aos procedimentos em humanos e avaliar a recuperação de volume globular e proteínas plasmáticas totais nos doadores. Os procedimentos foram realizados com o equipamento Fresenius AS104, com duração média de 1h46min, processamento de 5758mL de sangue total e colheita média de 3133mL de plasma. Não foram observadas alterações significativas do volume globular após a plasmaférese automatizada. A recuperação dos níveis plasmáticos de proteínas foi de 91,4% em 96 horas após o procedimento. A plasmaférese automatizada apresentou-se viável para a espécie equina, diminuindo o tempo de recuperação hematimétrica nos doadores.

This paper aimed to study feasibility and standardize the automated plasmapheresis in five healthy horses, showing the complications during the procedure, adjustments in relation to the procedures in humans and assessing the recovery of globular volume and plasma total proteins in donors. The procedures were performed with the Fresenius AS104 equipment, with an average duration of one hour and forty six minutes, processing 5758mL of whole blood and harvest average of 3133mL of plasma. There were no significant variations in globular volume after the automated plasmapheresis. The recovery of plasma total proteins was 91.4% at 96 hours after the procedure. The automated plasmapheresis appeared viable for the equine species, decreasing the time of hematimetric level recovery in donors.

A Indústria avícola brasileira cresce anualmente e se torna cada vez mais representativa na produção e exportação dos seus produtos. Os cuidados com a sanidade avícola têm acompanhado e favorecido essa evolução, entretanto, agentes respiratórios que afetam o peso e a qualidade da carcaça, continuam a provocar grandes prejuízos à produção avícola. A aerossaculite é considerada uma das principais causas da condenação total e/ou parcial de carcaças de frangos de corte, sendo de grande importância Mycoplasma gallisepticum (MG). O objetivo do presente estudo foi detectar MG pela PCR e correlacionar sua positividade a aerossaculite, queda de peso e condenação de carcaças de lotes de frangos de corte na Inspeção Sanitária Federal. Do total de 40 lotes de frangos de corte abatidos sob Inspeção Sanitária Federal, localizado no Estado do Rio Grande do Sul, Brasil, foram selecionados ao acaso. Em cada lote, três frangos de corte, independente de sexo, foram randomicamente selecionados para necropsia, sendo as traquéias coletadas e agrupadas em pool para formação de uma amostra para análise. Pela PCR, o DNA foi extraído pelo método de fenol-clorofórmio e amplificado com pares de "primers" específicos para MG. Dos 40 lotes analisados pela PCR, 20% (8/40) foram positivos para MG. Houve relação entre a positividade para MG, aumento da taxa de aerossaculite e queda de peso por Regressão Logística Múltipla (p<0,05), LogitPi= 7,9409 + (0,5601 x X1) - (3,3080 x X2). O aumento da taxa de aerossaculite esteve relacionada à queda de peso por Regressão Linear Simples (p<0,05), Y= 2, 1050-0,6397X. Em conclusão, a positividade por MG está relacionada à aerossaculite que provoca queda de peso em frangos de corte. Em adição, a PCR foi uma técnica eficaz para a detecção de MG em lotes de frangos de corte, não sendo este diagnóstico influenciado pelo tipo de colheita do material biológico, por escarificado ou swab de traquéia.

The Brazilian poultry industry improves annually and is more representative on production and exportation of their products. Care on poultry health cooperates to these developments, however, respiratory agents that affect weight and carcass quality, continue to threaten poultry production. Airsacculitis is considered the main cause of total and/or partial condemnation of broilers carcasses, being Mycoplasma gallisepticum (MG) the most important agent of it. This study aimed to detect MG by "Polymerase Chain Reaction" (PCR) and to correlate its positivity with airsacculitis, weight losses and condemnated broilers by Federal Sanitary Inspection. A total of 40 flocks of slaughter broilers under Federal Inspection in Rio Grande do Sul, Brazil, were randomly selected. In each flock three broilers, regardless of sex, were randomly chosen for necropsy where tracheas were collected and polled in one sample for analysis. For PCR, DNA was extracted by the method of phenol-chloroform and amplified with pairs of specific primers for MG. From the 40 flocks PCR analyzed, 20% (8/40) were positive for MG. MG detection was found to be correlected with airsacculitis increase and weight decrease by multiple logistic regression equation (p<0,05), LogitPi= 7.9409 + (0,5601 x X1) - (3.3080 x X2). Airsacculitis rate increase also correleted with decrease in body weight by simple linear regression equation (p<0.05), Y= 2.1050 - 0.6397X. In conclusion, MG positivity is related to airsacculitis which causes weight loss in broilers. In addition, the PCR was an effective technique for the detection of MG in flocks of broilers, but was not affected by the kind of biological specimens collected, as tracheal scraping or swab.

Neste experimento, foi definida uma combinação de sondas fluorescentes: iodeto de propídio (PI), aglutinina de Pisum sativum conjugada ao isotiocionato de fluoresceína (FITC-PSA) e JC-1. Para esta proposta, quatro ejaculados de três carneiros (n=12), que apresentavam motilidade >80% e alterações morfológicas <10%, foram diluídos em meio TALP e divididos em duas alíquotas. Uma alíquota foi submetida a três ciclos de flash frozen e descongelação, para induzir danos nas membranas celulares e distúrbios na função mitocondrial. Três tratamentos foram preparados com as seguintes proporções preestabelecidas de sêmen fresco: sêmen submetido a flash frozen: 0:100 (T0), 50:50 (T50) e 100:0 (T100). As amostras foram coradas no protocolo proposto e avaliadas por microscopia de epifluorescência. Para integridade de membrana plasmática, detectada pela sonda PI, foi obtida a equação: v=1,09+0,86X (R²=0,98). O acrossomo intacto, verificado pela sonda FITC-PSA, produziu a equação: v=2,76+0,92X (R²=0,98). O alto potencial de membrana mitocondrial, marcada em vermelho-alaranjado pelo JC-1, foi estimado pela equação: v=1,90+0,90X (R²=0,98). As equações lineares resultantes demonstraram que a técnica é eficiente e prática para avaliação simultânea das membranas plasmática, acrossomal e mitocondrial em espermatozoides de carneiros.

In this experiment, it was defined a protocol of fluorescent probes combination: propidium iodide (PI), fluorescein isothiocyanate-conjugated Pisum sativum agglutinin (FITC-PSA), and JC-1. For this purpose, four ejaculates from three different rams (n=12), all showing motility >80% and abnormal morphology <10%, were diluted in TALP medium and split into two aliquots. One of the aliquots was flash frozen and thawed in three continuous cycles, to induce damage in cellular membranes and to disturb mitochondrial function. Three treatments were prepared with the following fixed ratios of fresh semen:flash frozen semen: 0:100 (T0), 50:50 (T50), and 100:0 (T100). Samples were stained in the proposal protocol and evaluated by epifluorescence microscopy. For plasmatic membrane integrity, detected by PI probe, it was obtained the equation: v=1.09+0.86X (R²=0.98). The intact acrosome, verified by the FITC-PSA probe, produced the equation: v=2.76+0.92X (R²=0.98). The high mitochondrial membrane potential, marked in red-orange by JC-1, was estimated by the equation: v=1.90+0.90X (R²=0.98). The resulting linear equations demonstrate that this technique is efficient and practical for the simultaneous evaluations of the plasmatic, acrosomal, and mitochondrial membranes in ram spermatozoa.

O carbúnculo sintomático é causado pelo Clostridium chauvoei, enquanto o edema maligno é causado pelo C. chauvoei, C. septicum, C. sordellii, C. perfringens tipo A, e/ou C. novyi tipo A. Corpos policlonais anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C.novyi tipo A foram produzidos em coelhos e purificados em uma coluna de DEAE-celulose. Alíquotas das imunoglobulinas foram conjugadas com isotiocianato de fluoresceína e o restante foi usado na técnica de streptavidina biotina peroxidase (SBP). SBP foi padronizada para detectar C. chauvoei, C. septicum, C. sordellii e C. novyi tipo A em tecidos de cobaias fixados em formol e incluídos em parafina. A mesma foi comparada com a técnica de imunofluorescência direta (IFD). Secções e impressões do músculo da área de inoculação, coração, fígado, baço e rim, foram obtidas para ambas as técnicas. Nenhuma reação cruzada foi observada quando tecidos inoculados com outras espécies de Clostridia foram tratadas com esses quatro anticorpos primários. C. chauvoei e C. septicum foram detectados em todos os espécimes provenientes dos animais inoculados com esses microrganismos, enquanto somente secções de músculo obtidas de todos os animais inoculados com C. sordellii e C. novyi tipo A foram positivas. Os mesmos resultados observados pela técnica de SBP, foram obtidos em impressões de tecidos desses microrganismos corados pela IFD. Os resultados indicam que a técnica de SBP permite a detecção de C. chauvoei, C. septicum, C. sordellii e C. novyi tipo A em tecidos de cobaias fixados em formol e incluídos em parafina.

Blackleg is caused by Clostridium chauvoei, whereas malignant oedema is caused by C. chauvoei, C. septicum, C. sordellii, C. perfringens type A, and/or C. novyi type A. Anti-C. chauvoei, anti-C. septicum, anti-C. sordellii and anti-C. novyi type A polyclonal antibodies were produced in rabbits and purified in a column of DEAE-cellulose. Aliquots of the antisera were conjugated with fluorescein isothiocyanate and the remaining was used for the streptavidin biotin peroxidase technique (SBPT). SBPT was standardized to detect C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs. SBPT was compared to a fluorescent antibody technique (FAT). Sections and smears of muscle from inoculation area (MIA), heart, liver, spleen and kidney, were obtained for both SBPT and FAT. Cross-reactions between the different Clostridial species were not observed. C. chauvoei and C. septicum were detected in all specimens from the animals inoculated with these microorganisms, while only sections of muscle obtained from all the animals inoculated with C. sordellii and C. novyi type A were positive. The same results observed by the SBPT, were obtained on tissue smears of these microorganisms stained by the FAT. The results indicate that SBPT is suitable for detection of C. chauvoei, C. septicum, C. sordellii and C. novyi type A in formalin-fixed, paraffin-embedded tissues of guinea pigs.

São descritos surtos de enterotoxemia em caprinos em cinco propriedades no Estado do Rio Grande do Sul. Os animais afetados eram, normalmente, encontrados mortos ou apresentavam evolução aguda de 2 a 3 horas com acentuada depressão, cólicas abdominais e diarréia profusa com fibrina. Em duas propriedades relataram-se casos com a evolução de até 12 horas. Em treze animais necropsiados observaram-se aumento de líquidos nas cavidades abdominal, torácica e pericárdica, congestão e hiperemia da serosa e mucosa do intestino, conteúdo do cólon líquido com fibrina além de hemorragias de serosa e fibrina. Em um animal constatou-se microangiopatia cerebral caracterizada por acúmulo de material homogêneo e eosinofílico no espaço perivascular. No conteúdo intestinal, colônias com bastonetes morfológica e bioquimicamente sugestivos de Clostridium perfringens foram caracterizadas no estudo bacteriológico. A soroneutralização em camundongos com conteúdo intestinal dos animais afetados, revelou a presença da toxina épsilon. Estes achados evidenciam a enterotoxemia como doença de importância para criação de caprinos no Rio Grande do Sul.

Five outbreaks of caprine enterotoxaemia in Rio Grande do Sul, Brazil, were studied. The animals were found dead or had a clinical course that usually lasted 2-3 hours. From two farms a clinical manifestation period of 12 hours was reported. Clinical signs were characterized by depression, marked abdominal discomfort, profuse watery diarrhea with fibrin clots, and death. Thirteen necropsies were performed and hydropericardium, hydrothorax and hydroperitoneum were commonly found. The mucosa and serosa of the colon were congested, and its contents was watery with multiple fibrin clots. Serosal hemorrhages and fibrin clots in the gallbladder were also seen. Fibrinous colitis and thyphilitis were the most frequent histological changes. Cerebral microangiopathy was observed in one case, which was characterized by acute perivascular proteinaceous edema around arterioles of the nucleus caudatus and thalamus. Large numbers of Gram-positive rods, often in clumps and with morphology consistent with Clostridium perfringens, were seen in intestinal smears. The intestinal contents of six goats were positive for epsilon toxin by mouse neutralization test. These findings suggest that enterotoxaemia is an important disease of goat flocks in southern Brazil.

We studied the parasite fauna and food contents of 38 juvenile of Notothenia c.f. angustata specimens captured in the intertidal of Island Santa María and Maule, in the Arauco Gulf, Chile. Trophic habits of Notothenia c.f. angustata were not different between localities, which was mainly algae. A total 11 parasitic taxa among were found with the digenean Lecitasther macrocotyle, cestode larvae Tetraphyllidea and Diphyllidea and nematode Ascarophis sp. being the dominant taxa. The abundance of parasites was similar in the two localities, although the infracommunity richness was higher in fish from Maule. The most abundant parasite was L. macrocotyle in hosts from Isla Santa María, while those from Maule they were the Diphyllidea larvae and nematodes Ascarophis sp. The body length of the hosts was not a good predictor of the variations in abundance and richness at infracommunities, probably because of the early ontogenetic stage of fishes.