Morpho-histology of co-occurrence of somatic embryos, shoots, and inflorescences within a callus of Limonium 'Misty Blue'.

This is the first attempt to report the co-occurrence of somatic embryos, shoots, and inflorescences and their sequential development from stem cell niches of an individual callus mass through morpho-histological study of any angiosperm. In the presence of a proper auxin/cytokinin combination, precambial stem cells from the middle layer of a compact callus, which was derived from the thin cell layer of the inflorescence rachis of Limonium, expressed the highest level of totipotency and pluripotency and simultaneously developed somatic embryos, shoots, and inflorescences. This study also proposed the concept of programmed cell death during bipolar somatic embryo and unipolar shoot bud pattern formation. The unique feature of this research was the stepwise histological description of in vitro racemose inflorescence development. Remarkably, during the initiation of inflorescence development, either a unipolar structure with open vascular elements or an independent bipolar structure with closed vascular elements were observed. The protocol predicted the production of 6.6 ± 0.24 and 7.4 ± 0.24 somatic embryos and shoots, respectively, from 400 mg of callus, which again multiplied, rooted, and acclimatised. The plants' ploidy level and genetic fidelity were assessed randomly before acclimatisation by flow cytometry and inter simple sequence repeats (ISSR) marker analysis. Finally, the survivability and flower quality of the regenerated plants were evaluated in the field.

The pygidial gland secretion of the forest caterpillar hunter, Calosoma (Calosoma) sycophanta: the antimicrobial properties against human pathogens.

Recently, various insect secretions have been tested as possible antimicrobial agents. In beetles, these secretions are essentially products of various exocrine glands, of which particular emphasis is on pygidial glands that are common for the suborder Adephaga. The antimicrobial potential of the pygidial gland secretion isolated from the adults of Calosoma sycophanta against human pathogens has been tested and compared with the potential of other carabid species, particularly within the tribe Carabini. The antimicrobial assay includes a microdilution method which was applied in order to determine the minimal inhibitory, minimal fungicidal and minimal bactericidal concentrations. It has been tested the effect of the secretion against eight strains of fungal and eight strains of bacterial species. The secretion sample of the tested carabid species has shown statistically significant antifungal effect against all strains of treated micromycetes, the highest in comparison with previously tested carabids (Carabus spp., Laemostenus punctatus and Abax parallelepipedus). Aspergillus versicolor proved to be the most sensitive micromycete, while the remaining seven fungal strains have shown the same level of sensibility. In comparison with commercial mycotics ketoconazole and bifonazole, applied as positive controls, the tested secretion showed much higher antifungal activity for all fungal strains. Antibacterial effect has been manifested only against one bacterial strain (Escherichia coli), contrary to other previously studied carabid species. These observations might have a significant impact on the ecological domain and possible purpose in biomedical studies and applications in the future. Additionally, morpho-histology of the pygidial glands of C. sycophanta is investigated and discussed.

Antimicrobial activity of the pygidial gland secretion of three ground beetle species (Insecta: Coleoptera: Carabidae).

The antimicrobial properties of the pygidial gland secretions released by the adults of the three ground beetle species, Carabus ullrichii, C. coriaceus, and Abax parallelepipedus, have been tested. Microdilution method was applied for detection of minimal inhibitory concentrations (MICs), minimal bactericidal concentrations (MBCs), and minimal fungicidal concentrations (MFCs). Additionally, morpho-histology of the pygidial glands is investigated. We have tested 16 laboratory and clinical strains of human pathogens--eight bacterial both gram-positive and gram-negative species and eight fungal species. The pygidial secretion samples of C. ullrichii have showed the strongest antimicrobial effect against all strains of treated bacteria and fungi. Staphylococcus aureus, Lysteria monocytogenes, and Salmonella typhimurium proved to be the most sensitive bacterial strains. Penicillium funiculosum proved to be the most sensitive micromycete, while P. ochrochloron and P. verrucosum var. cyclopium the most resistant micromycetes. The pygidial secretion of C. coriaceus has showed antibacterial potential solely against Pseudomonas aeruginosa and antifungal activity against Aspergillus fumigatus, A. versicolor, A. ochraceus, and P. ochrochloron. Antibacterial properties of pygidial gland secretion of A. parallelepipedus were achieved against P. aeruginosa, while antifungal activity was detected against five of the eight tested micromycetes (A. fumigatus, A. versicolor, A. ochraceus, Trichoderma viride, and P. verrucosum var. cyclopium). Commercial antibiotics Streptomycin and Ampicillin and mycotics Ketoconazole and Bifonazole, applied as the positive controls, showed higher antibacterial/antifungal properties for all bacterial and fungal strains. The results of this observation might have a significant impact on the environmental aspects and possible medical purpose in the future.

Morpho-Histology and Morphometry of Chicken Testes and Seminiferous Tubules among Yellow-Feathered Broilers of Different Ages.

Unlike in many mammals, poultry testes are found in the abdominal cavity where they develop and perform spermatogenesis at high body temperature. Scarce reports among current publications detail the growth of testes and ST morphometry among juvenile chicks. Therefore, this study aims to investigate changes in components occurring in Gallus domesticus testes, by assessing the GSI and morphologically and histologically evaluating the testes and ST morphometry from 1-wk- to 4-mo-old. Right and left testes were collected from 70 healthy chickens divided into seven age-related groups (n = 10) and then immersed into the alcoholic acetate formalin (AAF) fixative solution. Hematoxylin- and eosin-stained tissues were used for microscopic observations. The findings revealed that both testes exhibited smooth features from 1-wk-old to 1-mo-old, and thereafter showed a consistent increase in vascularization until 4-mo-old. Histologically, both testes exhibited unclear ST, with ST apoptotic resorption observed in the 1-wk-old chicks. Until 1-mo-old, ST formed and few spermatogonia differentiated into primary spermatocytes, with all spermatogenic cells observed at 3-mo-old, i.e., sexual maturity. These findings suggest that both testes develop in analogy, and their sizes including increases in length and diameter are related to the spermatogenic activity in the ST. Subsequently, ST resorption by apoptosis is assumed to participate in the physiological mechanism regulating germ cells (GC). Finally, the GSI tended to increase with growth.

Seeds cryopreservation of Vriesea reitzii Leme & A.F. Costa endemic bromeliad from Atlantic Rainforest / Criopreservação de sementes de Vriesea reitzii Leme & A.F. Costa bromélia endêmica da Mata Atlântica

Vriesea reitzii is an endemic bromeliad from the Atlantic Rainforest. The objective of this research was to evaluate the cryopreservation using the method of direct immersion of its seeds, collected from capsules at 120, 135, and 150 days after anthesis (DAA). The water content was determined before cryopreservation, while the germination percentage, germination speed index (GSI), and total soluble carbohydrates were quantified after cryopreservation. The highest percentage of moisture (17.6%) was observed in 120 DAA, while the highest percentage of germination (89.6%) and GSI (17.0) were observed in 150 DAA. Optical and transmission electron microscopy analyses were performed, and no cell damage or changes at the morpho-histological and ultrastructural levels were observed after the cryopreservation process. From these results, V. reitzii seeds can be classified as orthodox seeds and the cryopreservation (+LN) is an efficient tool for an ex situ conservation of this species.

Vriesea reitzii é uma bromélia endêmica da Mata Atlântica. O objetivo deste trabalho foi avaliar a criopreservação de sementes utilizando o método de imersão direta, coletadas de cápsulas aos 120, 135 e 150 dias após a antese (DAA). O teor de água foi determinado antes da criopreservação. A porcentagem de germinação, o índice de velocidade de germinação (GSI) e os carboidratos solúveis totais foram quantificados após a criopreservação. A maior porcentagem de umidade (17,6%) foi observada aos 120 DAA, enquanto a maior porcentagem de germinação (89,6%) e GSI (17,0) foram observadas aos 150 DAA. Em análises de microscopia ótica e eletrônica de transmissão não foram observados danos celulares ou alterações a nível morfohistológico e ultraestrutural após o processo de criopreservação. A partir desses resultados, as sementes de V. reitzii podem ser classificadas como ortodoxas e a criopreservação (+LN) é uma ferramenta eficiente para a conservação ex situ desta espécie.