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One of the crucial aspects to be considered for successful in vitro production (IVP) of embryos is the composition of the various media used throughout the stages of this reproductive biotechnology. The cell culture media employed should fulfill the metabolic requirements of both gametes during oocyte maturation and sperm development, as well as the embryo during its initial cell divisions. Most IVP protocols incorporate blood serum into the media composition as a source of hormones, proteins, growth factors, and nutrients. Numerous studies have suggested Platelet-Rich Plasma (PRP) as a substitute for fetal sera in cell culture, particularly for stem cells. Therefore, the objective of this study is to assess the potential use of PRP as a replacement for fetal bovine serum (FBS) during oocyte maturation for in vitro production of bovine embryos. During in vitro maturation (IVM), cumulus-oocyte complexes (COCs) were allocated into the following experimental groups: Group G1 (IVM medium with 5% PRP); Group G2 (MIV medium with 5% PRP and 5% SFB); Group G3 (MIV medium with 5% SFB); and Group G4 (MIV medium without either PRP or SFB). Subsequently, the cumulus-oocyte complexes were fertilized with semen from a single bull, and the resulting zygotes were cultured for seven days. Cleavage and blastocyst formation rates were assessed on days 2 and 7 of embryonic development, respectively. The quality of matured COCs was also evaluated by analyzing the gene expression of HSP70, an important protein associated with cellular stress. The results demonstrated that there were no significant differences among the experimental groups in terms of embryo production rates, both in the initial cleavage stages and blastocyst formation (except for the G4 group, which exhibited a lower blastocyst formation rate on D7, as expected). This indicates that PRP could be a cost-effective alternative to SFB in the IVP of embryos.(AU)

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We evaluated the post-thaw development of in vitro-produced (IVP) bovine embryos cryopreserved using vitrification and slow-freezing techniques on different days after in vitro fertilization (IVF). Nine replicates of IVP were performed. Embryos at the expanded blastocyst stage with quality grades 1 and 2 (according to the International Embryo Technology Society (IETS) manual) were selected on days 7, 7.5, and 8 after IVF. Embryos (n = 472) were randomly divided and cryopreserved using slow freezing (n = 257) or vitrification (n = 215). The embryos were organized into six groups according to the cryopreservation technique and the day: 1) Group DT7 (embryos subjected to slow freezing on D7, n = 140); 2) Group DT7.5 (embryos subjected to slow freezing 12 hours after D7, on D7.5; n = 61); 3) Group DT8 (embryos subjected to slow freezing on D8, n = 56); 4) Group VIT7 (embryos vitrified on D7, n = 127); 5) Group VIT7.5 (embryos vitrified 12 hours after D7, on D7.5; n = 49); and 6) Group VIT8 (embryos vitrified on D8, n = 39). Data were arcsine-transformed and analyzed using analysis of variance and the GLIMMIX procedure in SAS (SAS 9.2), with P < 0.05. The re-expansion and embryo hatching rates were higher in vitrified embryos than in embryos subjected to slow freezing (P < 0.05). Embryo quality grade did not influence the total developmental rate (P > 0.05). However, grade 1 embryos re-expanded more rapidly (within 24 hours) than grade 2 embryos. Grade 1 embryos showed better results with vitrification than with slow freezing. The experimental groups represented a technique × day interaction and did not differ in terms of re-expansion and hatching rates (P > 0.05).(AU)

Avaliou-se o desenvolvimento pós-descongelamento de embriões bovinos produzidos in vitro (PIV) criopreservados pela técnica de vitrificação e de congelamento lento em diferentes dias após a fertilização in vitro (FIV). Foram realizadas 9 réplicas de PIV. Nos dias 7; 7,5 e 8 após a FIV, foram selecionados embriões no estágio de blastocisto expandido de grau de qualidade 1 e 2 (de acordo com o manual IETS). Os embriões (n=472) foram divididos aleatoriamente para serem criopreservados por congelamento lento (n=257) ou vitrificação (n=215). Organizou-se os embriões em 6 grupos de acordo com a técnica de criopreservação e o dia: 1) Grupo DT7 (embriões submetidos ao congelamento lento no D7, n= 140); 2) Grupo DT7,5 (embriões submetidos ao congelamento lento 12 horas após o D7, no D7,5; n= 61); 3) Grupo DT8 (embriões submetidos ao congelamento lento no D8, n=56); 4) Grupo VIT7 (embriões vitrificados no D7, n= 127); 5) Grupo VIT7,5 (embriões vitrificados 12 horas após o D7, no D7,5; n= 49); 6) Grupo VIT8 (embriões vitrificados no D8, n= 39). Os dados foram transformados para arcoseno e analisados por Análise de Variância utilizando o procedimento GLIMMIX do SAS (SAS 9.2) com P<0,05. A taxa de re-expansão e de eclosão embrionária foi maior para embriões vitrificados que para embriões submetidos ao congelamento lento (P<0,05). O grau de qualidade embrionária não influenciou as taxas de desenvolvimento totais (P>0,05). Porém embriões de grau 1 re-expandiram mais rapidamente (às 24 h) que embriões grau 2. Embriões grau 1 apresentaram melhores resultados com a técnica de vitrificação do que com congelamento lento. Os grupos experimentais representaram a interação técnica*dia e não diferiram entre si quanto as taxas de re-expansão e eclosão (P>0,05).(AU)

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Oviduct fluid extracellular vesicles (oEV) are essential for periconceptional events. The presence of EV has already been identified in the oviduct fluid (OF) from mammalian species, except in caprine. Therefore, this study aimed to isolate and characterize the caprine oEV (coEV). Initially, in Experiment 1, coEV were isolated from the OF of either each animal individually or from a pool of three animals. In experiment 2, coEV were isolated during the follicular or luteal phases of the estrous cycle. The coEV were characterized by size distribution, polydispersity index (PDI), and zeta potential using dynamic light scattering (DLS) analysis, as well as, by transmission electron microscopy (TEM) and dot blotting (DB). Our results indicated that the physicochemical characteristics of the coEV were similar (P > 0.05), regardless of the isolation method (individual or pool). However, coEV collected during the luteal phase were larger (P < 0.05) than those during the follicular phase. The TEM showed spherical and cup-shaped particles, characteristic of exosomes. The DB revealed the presence of exosomal proteins involved in the biogenesis of coEV. In conclusion, it is possible to isolate and characterize coEV from a single caprine female and the estrous cycle phase influences the vesicles average size and PDI.(AU)

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PICK1 plays a crucial role in mammalian spermatogenesis. Here, we integrated single-molecule long-read and short-read sequencing to comprehensively examine PICK1 expression patterns in adult Baoshan pig (BS) testes. We identified the most important transcript ENSSSCT00000000120 of PICK1, obtaining its full-length coding sequence (CDS) spanning 1254 bp. Gene structure analysis located PICK1 on pig chromosome 5 with 14 exons. Protein structure analysis reflected that PICK1 consisted of 417 amino acids containing two conserved domains, PDZ and BAR_PICK1. Phylogenetic analysis underscored the evolutionary conservation and homology of PICK1 across different mammalian species. Evaluation of protein interaction network, KEGG, and GO pathways implied that interacted with 50 proteins, predominantly involved in glutamatergic synapses, amphetamine addiction, neuroactive ligand-receptor interactions, dopaminergic synapses, and synaptic vesicle recycling, and PICK1 exhibited significant correlation with DLG4 and TBC1D20. Functional annotation identified that PICK1 was involved in 9 GOs, including seven cellular components and two molecular functions. ceRNA network analysis suggested BS PICK1 was regulated by seven miRNA targets. Moreover, qPCR expression analysis across 15 tissues highlighted that PICK1 was highly expressed in the bulbourethral gland and testis. Subcellular localization analysis in ST (Swine Tesits) cells demonstrated that PICK1 significantly localized within the cytoplasm. Overall, our findings shed new light on PICK1's role in BS reproduction, providing a foundation for further functional studies of PICK1.(AU)

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The effect of peptide toxins on voltage-gated ion channels can be reliably assessed using electrophysiological assays, such as the patch-clamp technique. However, much of the toxinological research done in Central and South America aims at purifying and characterizing biochemical properties of the toxins of vegetal or animal origin, lacking electrophysiological approaches. This may happen due to technical and infrastructure limitations or because researchers are unfamiliar with the techniques and cellular models that can be used to gain information about the effect of a molecule on ion channels. Given the potential interest of many research groups in the highly biodiverse region of Central and South America, we reviewed the most relevant conceptual and methodological developments required to implement the evaluation of the effect of peptide toxins on mammalian voltage-gated ion channels using patch-clamp. For that, we searched MEDLINE/PubMed and SciELO databases with different combinations of these descriptors: "electrophysiology", "patch-clamp techniques", "Ca2+ channels", "K+ channels", "cnidarian venoms", "cone snail venoms", "scorpion venoms", "spider venoms", "snake venoms", "cardiac myocytes", "dorsal root ganglia", and summarized the literature as a scoping review. First, we present the basics and recent advances in mammalian voltage-gated ion channel's structure and function and update the most important animal sources of channel-modulating toxins (e.g. cnidarian and cone snails, scorpions, spiders, and snakes), highlighting the properties of toxins electrophysiologically characterized in Central and South America. Finally, we describe the local experience in implementing the patch-clamp technique using two models of excitable cells, as well as the participation in characterizing new modulators of ion channels derived from the venom of a local spider, a toxins' source less studied with electrophysiological techniques. Fostering the implementation of electrophysiological methods in more laboratories in the region will strengthen our capabilities in many fields, such as toxinology, toxicology, pharmacology, natural products, biophysics, biomedicine, and bioengineering.

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ABSTRACT Blastocystis are common digestive tract parasites in humans and animals, extensively parasitic in humans and other primates. They exhibit extensive genetic diversity; Currently, 17 subtypes (STs) and some populations called non mammalian and avian STs (NMASTs) have been proposed. To understand the infection status and genotype distribution of Blastocystis sp. in wild mouse, this study used PCR technology to study the fecal DNA samples of 111 Leopoldomys edwardsi and 117 Berylmys bowersi collected from Guangdong and Chongqing. Among 228 fecal samples, 4 samples were positive for Blastocystis sp., with a total infection rate of 1.8% (4/228). Four positive samples formed two subtypes of ST3 and ST4, all of which were zoonotic genotypes. This article aims to investigate the infection status and genotype distribution of wild mouse Blastocystis sp., which will help reduce the infection of this pathogen to animals and thereby reduce their risk of zoonotic transmission.

RESUMO Blastocystis são parasitas comuns do trato digestivo em humanos e animais, amplamente parasitados em humanos e outros primatas. Atualmente, foram propostos 17 subtipos (STs) e algumas populações denominadas STs não mamíferos e aviários (NMASTs). Para entender o status da infecção e a distribuição do genótipo de Blastocystis sp. em camundongos selvagens, este estudo usou a tecnologia PCR para estudar as amostras de DNA fecal de 111 Leopoldomys edwardsi e 117 Berylmys bowersi coletadas em Guangdong e Chongqing. Entre 228 amostras fecais, 4 amostras foram positivas para Blastocystis sp. com uma taxa de infecção total de 1,8% (4/228). Quatro amostras positivas formaram dois subtipos de ST3 e ST4, todos os quais eram genótipos zoonóticos. Este artigo tem como objetivo investigar o status da infecção e a distribuição de genótipos de Blastocystis sp. em camundongos selvagens, o que ajudará a reduzir a infecção desse patógeno em animais e, assim, reduzir o risco de transmissão zoonótica.

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Mammalian prostate gland plays a role in alkaline substance synthesis including proteins. These functions are depending on glandular maturation and testosterone-androgen receptor (AR) dependent actions. Since tyrosine phosphorylated (TyrPho) proteins, also important for secreting pathways, have been localized in the androgen dependent organs, association between AR and TyrPho protein expressions in prostate is still unknown. This study aimed to investigate the changes of such proteins in prostate gland of male castrated rats. Nine prepubertal and adult twenty-two adult male rats were divided into the prepubertal (Pre, n=9), Sham (n=6), castrate for 3 (Cas-3, n=8) and for 7 (Cas-7, n=8) days groups, respectively. Serum testosterone level was determined. Histology and AR localization in each prostatic lobe were observed. TyrPho and AR protein expressions were also examined. The results showed undetectable testosterone level and low AR expression in Pre and Cas prostates with the decreased size. Few histopathologies were found in Cas groups. In ventral lobe, a Tyrpho protein was increased at the 48 kDa but the 52, 33, and 26 kDas were decreased in the Pre and Cas groups. For dorsolateral lobe, they were decreased at 33 and 30 kDas in Pre group and only 30 kDa was decreased in Cas-3 group. In the anterior lobe, the TyrPho proteins 57, 49, 39, 30, and 26 kDas were decreased in Pre group while 57, 30, and 26 kDas were decreased in Cas-3 group. In conclusion, the alterations of testosterone level and AR expressions associate with TyrPho protein expressions in prostate gland during development.

A próstata de mamíferos desempenha um papel na síntese de substâncias alcalinas, incluindo proteínas. Essas funções dependem da maturação glandular e das ações dependentes do receptor de testosterona-andrógeno (AR). Como as proteínas fosforiladas de tirosina (TyrPho), também importantes para as vias de secreção, foram localizadas nos órgãos dependentes de andrógeno, a associação entre as expressões de proteínas AR e TyrPho na próstata ainda é desconhecida. Este estudo teve como objetivo investigar as alterações dessas proteínas na próstata de ratos machos castrados. Nove ratos machos pré-púberes e adultos, vinte e dois adultos, foram divididos nos grupos pré-púberes (Pré, n=9), Sham (n=6), castrados por 3 (Cas-3, n=8) e por 7 (Cas-7, n=8) dias, respectivamente. O nível sérico de testosterona foi determinado. Histologia e localização de AR em cada lobo prostático foram observadas. As expressões de proteína TyrPho e AR também foram examinadas. Os resultados mostraram nível indetectável de testosterona e baixa expressão de AR em próstatas Pre e Cas com tamanho reduzido. Poucas histopatologias foram encontradas em grupos Cas. No lobo ventral, uma proteína TyrPho foi aumentada em 48 kDa, mas 52, 33 e 26 kDas foram diminuídos nos grupos Pre e Cas. Para o lobo dorsolateral, eles foram diminuídos em 33 e 30 kDas no grupo Pre e apenas 30 kDa foram diminuídos no grupo Cas-3. No lobo anterior, as proteínas TyrPho 57, 49, 39, 30 e 26 kDas foram diminuídas no grupo Pre, enquanto 57, 30 e 26 kDas foram diminuídas no grupo Cas-3. Em conclusão, as alterações do nível de testosterona e expressões de AR se associam às expressões da proteína TyrPho na próstata durante o desenvolvimento.

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Purpose: Artesunate (ART) has been implicated in regulating the many processes of liver injury, but its roles in liver regeneration still need to be illustrated. Methods: In the present study, ART was used to pretreat hepatocyte cell line NCTC1469 to study the effect of ART on hepatocyte proliferation in vitro. Furthermore, the potency of ART as a regimen to promote liver regeneration and restore liver function was evaluated following partial hepatectomy (PH) on C57BL/6 mice. Results: ART concentration-dependently promoted hepatocyte proliferation and reduced apoptosis. Cell cycle and Ki-67 immunohistochemical analyses demonstrated that ART supplementation promoted the proliferation of hepatocytes and accelerated liver regeneration. Our results provided evidence that ART improved liver function in a dose-dependent manner, as indicated by decreased serum alanine aminotransferase, aspartate aminotransferase, and increased albumin, and hepatocyte growth factor levels in PH mice. Meanwhile, ART promoted the PI3K/Akt/mTOR signaling in NCTC1469 cells and liver tissue of PH mice. In addition, PI3K inhibitor LY294002 blocked the promotion effect of ART on hepatocyte proliferation and cell cycle progression. Conclusion: ART promoted hepatocyte proliferation via activation of the PI3K/Akt/mTOR pathway, which was beneficial to liver regeneration of PH-induced liver injury.

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Thalassemia is one of the most important genetic haemolytic diseases that cause the breakdown of red blood cells (RBCs) in patients with ß-Thalassemia major. The body does not produce enough haemoglobin, which is an important part of RBCs. When there is not enough haemoglobin, RBCs do not function properly in the body, so the condition continues for short periods of time. The current studies aimed to determine the extent of the impact of ß-Thalassemia major on some hormonal variables in the serum of 80 patients (40 males and 40 females) aged between (1-15) years, in addition to 20 healthy children of the same age range and of both sexes, who were considered as a control group. The results of this study showed a significant increase in the concentration of erythropoietin (EPO) by 187% in the serum of patients with ß-Thalassemia major compared to healthy of both sexes, with an increase of 188 in males and 183% in females. The highest significant increase was in the age group of (11-15) years in males and females compared to healthy control. The results also showed a significant decrease in the concentration of hepcidin and growth hormones in the serum of patients with a decrease of 55 and 56% respectively compared to healthy individuals of both sexes, with a highest significant decrease of 56 and 59% in males, and 55 and 52% in females respectively. The highest significant decrease was in the age group of (11-15) years for both hormones in males and females compared to healthy control based on age groups and sex.(AU)

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The aims of this study were to determine the effect of rutin on in vitro maturation (IVM) of oocytes from in vitro-grown sheep secondary follicles and to analyze the possible involvement of the mammalian target of rapamycin (mTOR) pathway in IVM under the influence of rutin. Secondary follicles were cultured for 18 days in α-Minimum Essential Medium (α-MEM) supplemented with bovine serum albumin (BSA), insulin, glutamine, hypoxanthine, transferrin, selenium, ascorbic acid, and leptin (control medium: α-MEM+). Next, the follicles were evaluated for morphology, antrum formation, and follicular diameter, and the rate of fully grown oocytes (≥110 µm). Fully grown oocytes were submitted to IVM in Tissue Culture Medium 199 (TCM199) supplemented with fetal bovine serum (FBS), luteinizing hormone (LH), and recombinant follicle-stimulating hormone (rFSH) (IVM control medium), or in this medium with 0.1, 1, or 10 µg mL-1rutin. At the end of IVM, the oocytes were evaluated for mitochondrial activity, the reactive oxygen species (ROS) and glutathione (GSH) levels, the percentage of meiotic resumption, DNA fragmentation, and mTOR pathway involvement. After 18 days of in vitro culture, 77.5% of the follicles were normal and 77.7% became antral follicles with a diameter of 380.41 µm. Furthermore, almost 70% of the oocytes grew in vitro, reaching a diameter ≥110 µm and were submitted to IVM. Supplementation with 10 µg mL-1rutin significantly increased the percentage of oocytes that resumed meiosis (47.27%) compared with the control medium (30.43%). There was a significant increase in the ROS and GSH levels in oocytes matured with 0.1 µg mL-1 rutin compared with the other rutin treatments (p < 0.05). Furthermore, oocytes matured with TCM199+ showed a higher (p < 0.05) percentage of DNA fragmentation (30%) than those that matured with 10 µg mL-1 rutin (0%). After IVM, oocytes matured in the presence or absence of rapamycin showed a similar percentage of meiotic resumption (61.76% for TCM199+ 10 µg mL-1 rutin and 70.73% for TCM + 10 µg mL-1 rutin + rapamycin; p > 0.05). In conclusion, supplementation with 10 µg mL-1 rutin increased meiosis resumption and reduced DNA damage.(AU)

Os objetivos deste estudo foram verificar o efeito da rutina sobre a maturação in vitro (MIV) de oócitos provenientes de folículos secundários de ovelhas cultivados in vitro e analisar o possível envolvimento da via mTOR na MIV, sob influência da rutina. Os folículos secundários foram cultivados por 18 dias em meio α-Mínimo Essencial (α-MEM) suplementado com albumina sérica bovina (BSA), insulina, glutamina, hipoxantina, transferrina, selênio, ácido ascórbico e leptina (meio controle: α-MEM+). Em seguida, os folículos foram avaliados quanto à morfologia, formação do antro e diâmetro folicular e taxa de oócitos totalmente crescidos (≥110 µm). Oócitos totalmente crescidos foram submetidos à MIV em meio de cultivo de tecidos 199 (TCM199) suplementado com soro fetal bovino (FBS), hormônio luteinizante (LH), hormônio folículo estimulante recombinante (rFSH) (meio controle MIV) ou neste meio com 0,1, 1 ou 10 µg.mL-1 de rutina. Ao final da MIV, os oócitos foram avaliados quanto à atividade mitocondrial, concentração de espécies reativas de oxigênio (ERO) e glutationa (GSH), porcentagem de retomada de meiose, fragmentação de DNA e envolvimento da via mTOR. Após 18 dias de cultivo in vitro, 77,5% dos folículos estavam normais e 77,7% tornaram-se folículos antrais, com 380,41 µm de diâmetro. Além disso, 70% dos oócitos que cresceram in vitro atingiram diâmetro ≥110 µm e foram submetidos à MIV. A concentração de 10 µg.mL-1 de rutina aumentou significativamente a porcentagem de oócitos que retomaram a meiose (47,27%) em comparação ao meio controle (30,43%). Houve um aumento significativo nas concentrações de ROS e GSH em oócitos maturados com 0.1 µg.mL-1 de rutina em comparação com os outros tratamentos com rutina (p < 0,05). Além disso, a maturação de oócitos em TCM199+ aumentou (p<0,05) o percentual de fragmentação de DNA (30%) comparado ao tratamento com 10 µg.mL-1 de rutina (0%). Após MIV, ambos os tratamentos maturados na presença ou ausência de rapamicina apresentaram porcentagem semelhante de retomada meiótica (61,76% para TCM199+10 µg.mL-1 de rutina e 70,73% para TCM199+ 10 µg.mL-1 de rutina + rapamicina) (p>0,05). Em conclusão, a concentração de 10 µg.mL-1 de rutina aumentou a retomada da meiose e reduziu os danos ao DNA.(AU)

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Rabies is a viral encephalitis that affects mammals, including humans. Rapid and effective laboratory diagnosis of the rabies virus is critical for public health. This study evaluated the operational, technical, and financial viability of the RT-qPCR in replacement of inoculation in mice for diagnosing rabies in the laboratory routine. A total of 316 samples of mammalian brains were analyzed, 121 positives and 195 negatives, previously diagnosed by direct immunofluorescence (DIF) and mouse inoculation test (MIT). The samples were submitted to the duplex TaqMan RT-qPCR technique. The accuracy, sensitivity, and specificity of RT-qPCR were analyzed. We analyzed the costs for performing the RT-qPCR technique and compared it with the cost of MIT. The results showed 99.37% accuracy, 99.17% sensitivity, and 99.49% specificity by RT-qPCR when related to DIF and MIT results, which proved to be a robust and repeatable technique. The minimum time for a positive diagnosis was reduced in RT-qPCR (1 day) if compared to MIT (9.64 days), with a 17.7% reduction in the cost of the molecular technique. The present study demonstrated that the molecular biology technique is an efficient tool to diagnose rabies in the laboratory routine, being able to replace MIT.

A raiva é uma encefalite viral que acomete os mamíferos, incluindo humanos. O diagnóstico laboratorial rápido e eficaz do vírus da raiva é importante para a saúde pública. Este estudo teve como objetivo avaliar a viabilidade operacional, técnica e financeira da RT-qPCR em substituição à inoculação em camundongos para o diagnóstico da raiva na rotina laboratorial. Foram analisadas 316 amostras de cérebros de mamíferos, 121 positivas e 195 negativas, previamente diagnosticadas por imunofluorescência direta (IFD) e isolamento viral em camundongos, por inoculação intracerebral em camundongos. As amostras foram submetidas à técnica duplex TaqMan RT-qPCR. A precisão, sensibilidade e especificidade da RT-qPCR foram analisadas. Os custos para realizar a técnica de RT-qPCR foram analisados e comparados com o custo do isolamento viral em camundongos. Os resultados demostraram 99,37% de acurácia, 99,17% de sensibilidade e 99,49% de especificidade pela RT-qPCR quando relacionada aos resultados de IFD e isolamento viral em camundongos, que se mostrou uma técnica robusta e com repetibilidade. O tempo mínimo para diagnóstico positivo foi reduzido na RT-qPCR (1 dia) quando comparado ao isolamento viral em camundongos (9,64 dias), e com redução de 17,7% no custo pela técnica molecular. O presente estudo demonstrou que a técnica de biologia molecular é uma ferramenta eficiente para o diagnóstico da raiva na rotina laboratorial, podendo substituir o isolamento viral em camundongos.

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Investigating the interplay of factors that result in a viral zoonotic outbreak is difficult, though it is increasingly important. As anthropogenic influences shift the delicate balance of ecosystems, new zoonoses emerge in humans. Sub-Saharan Africa is a notable hotspot for zoonotic disease due to abundant competent mammalian reservoir hosts. Furthermore, poverty, corruption, and an overreliance on natural resources play considerable roles in depleting biological resources, exacerbating the population's susceptibility. Unsurprisingly, viral zoonoses have emerged in Africa, including HIV/AIDS, Ebola, Avian influenza, Lassa fever, Zika, and Monkeypox. These diseases are among the principal causes of death in endemic areas. Though typically distinct in their manifestations, viral zoonoses are connected by underlying, definitive factors. This review summarises vital findings on viral zoonoses in Africa using nine notable case studies as a benchmark for future studies. We discuss the importance of ecological recuperation and protection as a central strategy to control zoonotic diseases. Emphasis was made on moderating key drivers of zoonotic diseases to forestall future pandemics. This is in conjunction with attempts to redirect efforts from reactive to pre-emptive through a multidisciplinary "one health" approach.

Investigar a interação de fatores que resultam em um surto zoonótico viral é difícil, embora seja cada vez mais relevante. À medida que as influências antropogênicas mudam o delicado equilíbrio dos ecossistemas, novas zoonoses surgem em humanos. A África Subsaariana é um ponto crítico notável para doenças zoonóticas devido a abundantes reservatórios mamíferos competentes. Além disso, a pobreza, a corrupção e o excesso de confiança nos recursos naturais desempenham papéis consideráveis no esgotamento dos recursos biológicos, exacerbando a suscetibilidade da população. Sem surpresa, zoonoses virais surgiram na África, incluindo HIV/AIDS, Ebola, gripe aviária, febre de lassa, zika e varíola dos macacos. Essas doenças estão entre as principais causas de morte em áreas endêmicas. Apesar de serem tipicamente distintas em suas manifestações, as zoonoses virais estão conectadas por fatores subjacentes e definitivos. Esta revisão resume descobertas vitais sobre zoonoses virais na África usando nove estudos de caso notáveis como referência para estudos futuros. Discutimos a importância da recuperação e proteção ecológica como estratégia central para o controle de doenças zoonóticas. Foi dada ênfase à moderação dos principais impulsionadores de doenças zoonóticas para prevenir futuras pandemias. Isso ocorre em conjunto com tentativas de redirecionar os esforços de reativos para preventivos por meio de uma abordagem multidisciplinar de "uma só saúde".

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Quarenta e nove ovos embrionados de Psittaciformes com embriões que morreram durante a incubação foram examinados, provenientes de criatório comercial de aves de espécies exóticas ou nativas, ou provenientes de instituição de conservação de espécies da avifauna. Os ovos foram classificados, de acordo com o momento da morte, em mortalidade precoce, intermediária ou tardia. Conforme a idade embrionária, embriões inteiros ou tecidos embrionários foram coletados para extração de DNA e cultivo bacteriológico em ágar contendo acetato de tálio, soro equino e penicilina. Entre os embriões de espécies exóticas, 37,5% (12/32) foram detectados positivos para Mycoplasma spp. Considerando os embriões das espécies nativas, 52,4% foram detectados positivos (11/21). O DNA de Mycoplasma spp. foi detectado em um filhote de Pionus maximiliani morto na eclosão. Testes adicionais dos embriões e das colônias por PCR, com protocolo específico para M. gallisepticum, não revelaram nenhum resultado positivo. As implicações da presença de Mycoplasma na viabilidade embrionária e de filhotes de espécies de aves comerciais ou de conservação são discutidas.

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This study described changes in the serum biochemistry, morphology of genital organs, long bone, and eggshell during the daily egg formation cycle in Japanese quails. Sixty quails (18-wk) were distributed in 6 groups according to hours post-oviposition (POV): 0 hr POV (16h00), 2 hrs POV (egg in magnum), and 4, 8, 14, and 20 hrs POV (egg in uterus). The magnum had higher relative weight before the next ovulation (20 and 0 hr POV), and its tubular glands showed functional variation through periods: abundant eosinophilic, PAS+, and negative Alcian blue secretion at 0 and 2 hrs, empty glands aspect at 4 hrs, and filled again at 20 hrs POV. Serum albumin and total Ca had the highest value in the 2 hrs group, and the lowest in 8 and 14 hrs groups. Egg-cycle period affected the Ca% of the medullar bone of the femur and tibiotarsus, with the lowest mean at 14 hrs POV (06h00), and the highest mean after oviposition (0 hr POV), showing the recovery of Ca stores in long bones for the next egg cycle. Analysis of the eggshell using scanning electron microscopy evidenced that palisade layer formation starts during the night (8­14 hrs POV), and most parts are secreted during the day period. In conclusion, eggshell secretion in light periods, high magnum activity and medullary bone Ca deposition during midday and afternoon, as well as the ovulation/oviposition in the afternoon, are the main characteristics of the distinct physiological aspects of the egg cycle in quails.(AU)

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In xeric environments, such as Caatinga Biome, habitat characteristics such as phytophysiognomy type and presence of water bodies can represent higher resource availability. In this context, the present study investigated the effect of phytophysiognomies and presence of water bodies in the abundance and community structure of medium and large mammal species (MLM) in the Serra de Santa Catarina, Paraíba, Brazil. To evaluate these variables we conduct an effort of 373 camera-trap days, between August 2012 and November 2014. We recorded 12 MLM species, distributed in six orders and 11 families. From those, Kerodon rupestris is the only one listed in the Brazilian List of Threatened Fauna. Regarding the habitat, the Mann-Whitney showed a significant higher frequency to the Shrubby habitat and the ANOSIM showed no shifts in the community structure between Arboreal and Shrubby. Concerning the presence of water bodies, both the Mann-Whitney and the ANOSIM showed significant higher frequency to the habitat with water presence. We observed that both phytophysiognomy and water bodies are important variables which affect mainly the abundance of mammalian species from semiarid environments. Nonetheless, whereas the forest remnants get smaller the existence of water bodies becomes a preponderant factor to the MLM species and its community structure.(AU)

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RFX2 plays critical roles in mammalian spermatogenesis and cilium maturation. Here, the testes of 12-month-old adult boars of Banna mini-pig inbred line (BMI) were subjected to whole-transcriptome sequencing. The results indicated that the average expression (raw count) of RFX2 gene in BMI testes was 16138.25, and the average expression value of the corresponding transcript ENSSSCT00000043271.2 was 123.1898. The CDS of RFX2 obtained from BMI testes was 2,817 bp (GenBank accession number: OL362242). Gene structure analysis showed that RFX2 was located on chromosome 2 of the pig genome with 19 exons. Protein structure analysis indicated that RFX2 contains 728 amino acids with two conserved domains. Phylogenetic analysis revealed that RFX2 was highly conserved with evolutionary homologies among mammalian species. Other analyses, including PPI networks, KEGG, and GO, indicated that BMI RFX2 had interactions with 43 proteins involving various functions, such as in cell cycle, spermatid development, spermatid differentiation, cilium assembly, and cilium organization, etc. Correlation analysis between these proteins and the transcriptome data implied that RFX2 was significantly associated with FOXJ1, DNAH9, TMEM138, E2F7, and ATR, and particularly showed the highest correlation with ATR, demonstrating the importance of RFX2 and ART in spermatogenesis. Functional annotation implied that RFX2 was involved in 17 GO terms, including three cellular components (CC), six molecular functions (MF), and eight biological processes (BP). The analysis of miRNA-gene targeting indicated that BMI RFX2 was mainly regulated by two miRNAs, among which four lncRNAs and five lncRNAs competitively bound ssc-miR-365-5p and ssc-miR-744 with RFX2, respectively. Further, the dual-luciferase report assay indicated that the ssc-miR-365-5p and ssc-miR-744 significantly reduced luciferase activity of RFX2 3'UTR in the 293T cells, suggesting that these two miRNAs regulated the expression of RFX2. Our results revealed the important role of RFX2 in BMI spermatogenesis, making it an intriguing candidate for follow-up studies.(AU)

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The identification of biodiversity conservation priority sectors that are not formally protected, have an essential part of conservation strategies and goals at global and local scale. Ecological niche modeling is a relevant and important tool for analysis and distribution of species, also is used to determine the biodiversity patterns through the regions. The main objective of this work was to identify the sites with high biodiversity patterns in a sector of Chaco Seco ecoregion that haven't been protected with environmental legislation. Through biodiversity sampling with foot transects, camera traps and interviews, it was registered the presence of large and medium mammals in Santiago del Estero Province. Biodiversity pattern maps were then developed from potential distribution models (SDMs) of 5 mammalian species selected for being relevant for conservation. To define zones that could be characterized like conservation gaps, pattern maps were contrasted with protected areas layers and legal schemes of land use planning and also, protected forest. For the SDM, 171 records were used, 43 for M. gouazoubira, 40 for P. concolor, 20 for M. trydactila, 43 on P. tajacu and 25 on C. wagneri. Three models were used to make the biodiversity patterns, one of these, Fuzzy union, were used for the subsequent calculation. The total area of high biodiversity increases to 39.486 km², which represents the 29% of provincial area. In consequence the 81% remaining represents the conservation gaps areas for that sector of the ecoregion.

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Abstract Brazil is the world's richest country in biodiversity, including mammal species. In the Brazilian Cerrado biome, mammalian diversity is vast, with about 251 species, 32 of them are endemic and 22 listed as threatened species. In this work, we investigated species diversity of medium- and large-sized mammals in the private protected area RPPN Pontal do Jaburu (RPPN-PJ) and its surroundings, which is a flooded area located in an important biological corridor in the Cerrado-Amazon ecotone zone, a priority area for biodiversity conservation in Brazil. We used camera-trapping, active search (night and day), and track survey during dry season (Apr Aug 2016). We recorded 29 mammal species, being the Carnivora order the most representative with 11 species. Regarding threat status, 35.7% of the recorded species were listed as threatened in Brazil and 32.1% worldwide. We highlight the high relative frequency of threatened species records such as Tapirus terrestris, Panthera onca, Blastocerus dichotomus, Pteronura brasiliensis, Priodontes maximus, and other, as well as the presence of the newly described aquatic mammal species Inia araguaiaensis. We stress the importance of RPPN-PJ and its surroundings for mammal conservation, which include complex habitats (wetlands) located in an important ecotone zone.

Resumo O Brasil é o país mais rico em biodiversidade no mundo, incluindo espécies de mamíferos. No bioma Cerrado, a diversidade de mamíferos é enorme, com cerca de 251 espécies, sendo 32 delas endêmicas e 22 listadas como ameaçadas de extinção. Neste estudo, investigamos a diversidade de espécies de mamíferos de médio e grande porte da RPPN Pontal do Jaburu (RPPN-PJ) e seu entorno, que é uma floresta de inundação localizada em um importante corredor biológico na zona de ecótono Cerrado-Amazonia, uma área prioritária para conservação da biodiversidade no Brasil. Os dados foram coletados por armadilhas fotográficas, busca ativa (noturna e diurna) e identificação de pegadas durante a estação seca (abril - agosto de 2016). Registramos um grande número de espécies de mamíferos (n = 29), sendo a ordem carnívora a mais representativa com 11 espécies. Com relação ao status de ameaça, 34,5% das espécies registradas foram listadas como ameaçadas na lista vermelha do Brasil e 20,7% na lista vermelha da IUCN. Destacamos a alta frequência relativa de registros de espécies ameaçadas como Tapirus terrestris, Panthera onca, Blastocerus dichotomus, Pteronura brasiliensis, Priodontes maximus, bem como a presença da recém descrita espécie de mamífero aquático Inia araguaiaensis. Nós discutimos a importância da RPPN-PJ e seus arredores para a conservação de espécies de mamíferos, onde inclui habitats complexos (áreas de inundação) localizados em uma importante zona de ecótono.. Os resultados reforçam a relevância desta área para a conservação de mamíferos.

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O tecido testicular contém células germinativas, que possuem potencial para se desenvolver em espermatozoides viáveis por meio do cultivo in vitro ou xenotransplante, sendo uma alternativa interessante a ser utilizada na formação de biobancos. Esta revisão compila as atualizações, desafios e perspectivas relacionadas às técnicas de criopreservação e cultivo de tecido testicular como estratégia para a conservação de espécies mamíferas. O tecido testicular pode ser obtido tanto de indivíduos adultos como pré púberes, seja após orquiectomia ou até mesmo após a sua morte. O tecido fragmentado pode ser criopreservado por congelação lenta ou rápida e por métodos de vitrificação. Os crioprotetores são indispensáveis durante a criopreservação e podem variar o tipo e concentração de acordo com a espécie. Com os avanços da criopreservação deste material, espermatozoides podem ser obtidos por transplante de fragmentos testiculares ou células germinativas isoladas em camundongos imunodeficientes. No entanto, a obtenção de espermatozoides no cultivo in vitro ainda é um desafio.(AU)

The testicular tissue contains germ cells, which have the potential to develop into viable spermatozoa through in vitro culture or xenotransplantation, being an interesting alternative to be used in the formation of biobanks. This review compiles updates, challenges and perspectives related to cryopreservation techniques and testicular tissue culture as a strategy for the conservation of mammalian species. Testicular tissue can be obtained from both adult and pre-pubertal individuals, either after orchiectomy or even after their death. Fragmented tissue can be cryopreserved by slow or fast freezing and by vitrification methods. Cryoprotectants are indispensable during cryopreservation and may vary in type and concentration according to the species. With advances in cryopreservation of this material, spermatozoa can be obtained by transplanting testicular fragments or isolated germ cells into immunodeficient mice. However, obtaining spermatozoa in in vitro culture is still a challenge.(AU)

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Plastic pollution in our environment is one of the most important global health concerns right now. Micro- and nanoplastics (MNPs) are taken up by both humans and animals, mainly via food and water, and can pass important epithelial barriers. Indications of plastics in the blood circulation have recently been shown in both humans and farm animals, but standardized methods to quantify the exact levels of MNPs to which we are exposed are currently lacking. Potential hazards of MNPs are being investigated very recently, including the impact that MNPs may have on reproduction. However, studies on mammalian reproduction are scarce, but a wealth of data from aquatic species indicates reproductive effects of MNPs. The first studies in rodent models demonstrate that MNPs reach the gonads after oral exposure and may impact offspring after maternal exposure during the gestational period. These effects may arise from the particles themselves or the presence of plastic contaminants that leach from plastics. Plastic contamination has been detected in human placentas, fetal fluid and the meconium of newborns, indicating the presence of plastics from the very first start of life. Currently there is a lack of studies that investigate the impact of MNP exposure during the periconception and embryonic period, whereas this is an extremely sensitive period that needs considerable attention with the growing amount of plastics in our environment.(AU)