... Histological analyses were done by first cutting the organ mass through the mid-section containing the digestive gland and gonad. Tissue samples were fixed in alcoholic Bouin fluid, and processed using standard histological techniques . After dehydration in an ethanol series and clearing with Histosol Clearing Agent, the sections were embedded in Paraplast wax. ...
... After dehydration in an ethanol series and clearing with Histosol Clearing Agent, the sections were embedded in Paraplast wax. Tissue sections (6 µm thick) were stained with a trichrome stain , which included Alcian blue (8GX Sigma-Aldrich) at pH 2.5 to differentiate proteoglycans (blue granules). Gonad and digestive gland examination were done using a Leica DM2700 microscope. ...
Fighting conch Strombus pugilis is one of six Strombidae species distributed throughout the Caribbean. It is used as food, as an aquarium organism and its shell are popular in jewelry production. Conch aquaculture has been done traditionally by extracting egg masses from wild adults. This is an issue for several conch species protected by CITES. Intensive conch culture requires good growth rates and gonad maturity under laboratory conditions using formulated feed. An evaluation was done of the effect of inclusion of the red algae Halymenia and Spirulina on gonad maturity in S. pugilis using two experimental diets containing low and high concentrations of these algae (2% and 8% of each). Each diet was fed to six groups of conch kept in 20 L aquaria at 27.5°C. They were fed twice daily at 0.1 g feed/conch for 105 days. Gonad development and digestive gland structure were analyzed with histological techniques. Analysis of gonad development and vitellus granule diameter were analyzed for the two treatments and a control (wild conch). Wild conch females exhibited a reproductive cycle with 100% maturity at the beginning of this study, followed immediately by spawning (in two peaks: 50% and 34%) and initiation of a new oogenesis cycle. Females fed the 8% H. floresii and 8% Spirulina diet exhibited two spawning peaks (75% and 100%) spaced a month apart, and larger yolk granules than those in the control and the 2% H. floresii and 2% Spirulina diet. Proteoglycan granule abundance in the digestive cells did not differ between treatments. H. floressi and Spirulina may function as a feeding stimulant, enhancing feed intake and promoting gonadal maturity in S. pugilis broodstock under laboratory conditions.
... Other fragments were embedded for 1 min in Nile Blue, which is used to examine neutral and acidic lipids [Jensen 1962]. Moreover, the plant fragments were treated with Ruthenium Red for 5 min to determine polysaccharides other than cellulose [Johansen 1940], 1% potassium dichromate for 10 min to detect tannins, and 2% ferric trichloride for 10 min to identify phenolic compounds [Johansen 1940, Soloway and Wilen 1952, Gabe 1968. Furthermore, a reaction with concentrated sulphuric acid was performed to detect sesquiterpenes [Cappelletti et al. 1986]. ...
... flavonoids [Dewick 1977, Combrinck et al. 2007. Moreover, the potassium dichromate test [Gabe 1968] revealed the presence of tannins. These substances were found in chlorophorm and methanol extracts of the eyebright by Dimitrova et al. . ...
Micromorphological and anatomical investigations of trichomes on vegetative and generative organs of the drug eyebright (Euphrasia stricta D. Wolff ex J.F. Lehm.) were carried out. Additionally, identification of the main components of the secreted products was conducted. The following types of glandular structures were found: capitate trichomes (i) with a unicellular head, a neck cell, and a 2-celled stalk, (ii) with a unicellular head, a neck cell, and 3-celled stalk, (iii) with a bicellular head and sporadically with a 3- or 4-celled head; moreover, on the surface of corolla unicellular and 2-3-celled clavate trichomes, ribbon-like trichomes, and papillae were present. Using fluorescence microscopy and histochemical tests, various substances such as lipids, phenolic compounds, terpenes containing steroids, terpenoids, sesquiterpenes, tannins, and flavonoids were determined but only in the capitate trichomes. The analysis of the essential oil showed that its content in the dry herb of E. stricta was on average 0.257%. Gas chromatography revealed the presence of 28 compounds.
... All samples were finally dehydrated in butanol for 3 days, impregnated in two baths of 5 paraffin for two days and embedded in paraffin. They were sectioned transversely (cross-sections perpendicular to osteoderm keel) at 50 microns with a microtome and stained with a one-step trichrome or with orcein following Gabe's instructions ( Gabe, 1968). ...
... All samples were finally dehydrated in butanol for 3 days, impregnated in two baths of paraffin for two days and embedded in paraffin. They were sectioned transversely (cross-sections perpendicular to osteoderm keel) at 50 microns with a microtome and stained with a one-step trichrome or with orcein following Gabe's instructions ( Gabe, 1968). ...
Vascularization in the core of crocodylian osteoderms, and in their superficial pits has been hypothesized to be a key feature involved in physiological thermoregulation and/or acidosis buffering during anoxia (apnea). However, up to now, there have been no quantitative data showing that the inner, or superficial, blood supply of the osteoderms is greater than that occurring in neighboring dermal tissues. We provide such data: our results clearly indicate that the vascular networks in both the osteoderms and the pits forming their superficial ornamentation are denser than in the overlying dermis. These results support previous physiological assumptions and indicate that vascularization in pseudosuchian (crocodylians and close relatives) ornamented osteoderms could be part of a broad eco-physiological adaptation towards ectothermy and aquatic ambush predation acquired by the crocodylomorphs during their post-Triassic evolution. Moreover, regressions demonstrate that the number of enclosed vessels is correlated with the sectional area of the cavities housing them (superficial pits and inner cavities). These regressions can be used to infer the degree of vascularization on dry and fossilized osteoderms and thus document the evolution of the putative function of the osteoderms in the Pseudosuchia. This article is protected by copyright. All rights reserved.
... To study monthly gonad development and to measure oocyte diameter, the gonads were first embedded in paraffin, sectioned at 5-7 mm with a digital microtome (Leica), heated for 24 h at 608C and then finally stained with haematoxylin and eosin (Gabe 1968). Histological slides were observed and photographed under a Zeiss Axiostar light microscope. ...
Marine gastropods are a popular seafood, abundant along coastal shores and healthy for human consumption. We studied variations in nutritional components during the gonadal cycle of Tegula patagonica, a novel gastropod resource protected by a recent artisanal fishery regulation. Similar to other marine gastropods consumed worldwide, T. patagonica was found to have a high protein content (mean ± s.d., 28.8 ± 15.4%) and a low lipid (1.36 ± 0.43%) and carbohydrate (1.7 ± 0.8%) content. The spawning peak was registered during spring and summer, concordant with increases in temperature and photoperiod, as observed for other gastropods from Atlantic Patagonia. According to our results, the optimal quality of T. patagonica for consumption is during the austral autumn, after the spawning season and when the gastropods have a high bodyweight and best nutritional contribution. Thus, this resource could be captured when its quality is optimal and its conservation promoted if established minimum catch sizes are respected.
... Slices were cut and mounted on histological slides or on "Super Frost" glass slides for immunohistochemistry. After hydration with decreasing concentrations of ethanol, the sections were stained with Masson's trichrome and haematoxylin and eosin [17,41]. ...
Background: The efferent ducts are mainly involved in the reabsorption of the seminiferous tubular fluid. Testosterone and oestrogens regulate efferent ducts functions via their receptors. Materials and methods: This paper presents an experimental investigation on the location of the P450 aromatase, the 17-b oestradiol (E2), the androgen receptor (AR), the oestrogen receptor 1 (ESR1), the oestrogen receptor 2 (ESR2) and the G protein-coupled oestrogen receptor 1 (GPER1) in the efferent ducts using Psammomys obesus as an animal model to highlight the effect of the season on the histology and the distribution of these receptors. Results: We observed a proliferation of the connective tissue, decreasing in the height of the epithelium during the resting season compared to the breeding season. Ciliated cells expressed P450 aromatase, AR, E2, ESR1, ESR2 and GPER1 during both seasons. Basal cells showed a positive staining for the ESR1 and the GPER1 during both season, the AR and E2 during the breeding season and ESR2 during the resting season. Conclusions: Our result shows that the expression of androgen receptor and oestrogen receptors in the efferent ducts vary by season witch suggest that they are largely involved in the regulation of the efferent ducts functions. (Folia Morphol 2020; 79, 4: 756–766) Key words: G protein-coupled oestrogen receptor 1, oestrogen receptor, androgen receptor, aromatase, efferent ducts, sand rat.
... The general morphology and anatomy were studied by means of a stereo dissecting microscope. The anatomical and histological details were studied following the Ramon & Cajal method for topographic staining (see Gabe, 1968). Histology and nematocysts were examined with a light microscope. ...
The presence of some endemic taxa of the genus Actinia from Madeira and the Canary Islands is discussed. A new species, Actinia nigropunctata, is described, and some characters of the poorly known Actinia virgata are discussed. In addition, we discuss the main characters proposed by Schmidt (1971) to distinguish species of Actinia.
... For histological examination, a portion of the liver was fixed in 10% neutral buffered formalin embedded in paraffin, sectioned and stained with hematoxylin and eosin as described by Gabe . Then semi-thin sections (0.5-1 μm) were prepared by using LKB ultramicrotome. ...
Abstract Background: This study investigated whether the combination of sodium valproate (SV) with l-cysteine (LC) can decrease the SV toxicity of kidneys. SV caused alternation in oxidative/antioxidant balance. Methods: Biochemical estimations included the determination of oxidative stress markers like thiobarbituric acid-reactive substances in kidney tissue, and enzymatic antioxidant activities such as superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GPx), glutathione reductase as well as total antioxidant capacity were evaluated in renal tissues. Creatinine and uric acid levels in the serum were also determined to assess kidney function. Pathological examination of the kidney was also performed. Results: Increasing the levels of lipid peroxidation and decreasing the enzymatic activity (SOD, CAT, and GPx) as well as total antioxidant capacity of rats was shown with different doses of SV. Impairment in renal function tests suggests a decreased glomerular filtration rate, as serum creatinine was elevated. Histopathological changes of kidney tissue treated with SV reveal the proximal and the distal convoluted tubules that show hydropic changes (small white vacuoles within the cytoplasm and the glomeruli show hypercellularity). Conclusions: The concurrent administration of LC with SV significantly had beneficial effects on the kidney and all the above parameters have been improved.
... After measuring the oysters, they were anesthetised with clove oil (1.5 ml/L) for 10 min and the edge of the mantle was dissected. The samples were then fixed in Bouin's fluid ( Gabe 1968) for 48 h for the histological process, and then stored in 96% ethanol until analysis. ...
Shells in pearl oysters are produced by the mantle which is also used as a graft in pearl operations. Here, we investigate the mantle structure of a new pearl oyster species of the Persian Gulf, Pinctada persica, and compare it to two other pearl-producing species, Pinctada radiata and Pteria penguin. The anterior, ventral and posterior segments of the mantle edge of each valve were fixed, and tissue sections were stained with haematoxylin and eosin. A new pentachrome method and PAS-alcian blue staining were used to characterise the different mucosubstances. The mantle edges were found to have an outer, middle and inner fold, which have different morphology in each species. The mantle edge is lined by cuboidal and columnar epithelia, and interspersed among these epithelial cells we found mucous cells and cells that contained brown granules. The outer and middle folds of the two Pinctada species show different shapes to that of Pteria penguin. Most of the mucous cells in the mantle contain acidic mucosubstances and small amounts of mixed acidic-neutral mucosubstances were observed in the middle and inner fold of Pinctada persica. This study reveals that the mantle edges of the three species possess similar cellular structure, but vary in the shape of the folds. ARTICLE HISTORY
... Les trois paramètres, numération érythrocytaire (NGR), hémoglobine (Hb) et hématocrite (Ht), permettent de déterminer les constantes érythrocytaires : volume globulaire moyen [VGM = Ht/NGR x 10, en microncubes ([*)], concentration corpusculaire moyenne en hémoglobine [CCMH = Hb/H t X 100 ; en g/100 m] et taux globulaire moyen en hémoglobine [TGMH = Hb/NGR X 10, en picogramme (pg)]. Des étalements sanguins sur lames de verre ont été réalisés et colorés selon la méthode de Giemsa-Pappenheim (Gabe, 1968). Ils ont permis l 'identification des différentes variétés leucocytaires, et l 'établissement de leurs pourcentages respectifs (formule leucocytaire). ...
... Serial sectionsDOI 10.1007/s00360-017-1115-4 First Online: 10July 2017Springer Link to the online version of the paper: http://rdcu.be/t41u Prenant-Gabe's trichrome according to a classical protocol (Gabe 1968). Digital pictures and cell measurements of the 204 digestive gland were achieved with the Nikon C system combining Eclipse 80i microscope / DXM1200-C digital 205 camera and NIS-elements D 3.0 software.206The ...
In Sepia officinalis (Linnaeus, 1758), the digestive gland matures during the first month post-hatching while a shift from intracellular acid to extracellular alkaline digestion occurs. The purpose of this study was to investigate the possibility of using enzymatic ratios for the description of digestive system maturation in early life stages of S. officinalis. Secondly, it is intended to apply these new tools as eco-physiological indicators for understanding the impact of cuttlefish eggs’ life history from different spawning sites of the English Channel on digestive performance of juveniles. An experimental rearing was performed over 35 days after hatching (DAH) on juveniles from wild collected eggs in 2010 and 2011. Four digestive enzyme activities and their ratios [i.e. trypsin, cathepsin, acid (ACP) and alkaline (ALP) phosphatase, ALP/ACP and trypsin/cathepsin] were studied along with histological features [e.g. internal yolk surface and digestive gland development]. The two enzyme ratios were good indicators of digestive system maturation allowing the study of the digestive gland’s development. They were highly correlated to juveniles’ weight increase and histological features of the gland in early DAH. These ratios described more accurately the shift occurring between the intracellular acid and the extracellular alkaline modes of digestion in S. officinalis and were more specific than separated enzyme activities. Their application as eco-physiological tools revealed that enzyme ratios reflected yolk content and digestive gland development in new hatching juveniles. Finally, ALP/ACP ratio was shown to be a powerful tool to describe growth performance of S. officinalis which is useful for aquaculture optimization.
... The extracted specimens were transported to the laboratory where they were dissected and sections of the middle area of the gonad–hepatic gland complex of 100% of the females (considering that they better represent the reproductive processes (Cantillánez et al., 2011)), were fixed in Bouin and included in paraffin. Cuts 6 µm thick were colored with hematoxylin and eosin (Gabe, 1968), and were then microscopically analyzed to classify their maturity state, according to the scale proposed byCantillánez et al. (2011). ...
The reproductive condition of Thais chocolata (copulative aggregation establishment, clutch laying in capsules and specimen density), was studied monthly between February 2009 and January 2010, in the locality of Chanavaya (Tarapacá, Chile). Results showed that the specimens are mainly distributed in aggregations, and that their gonad development is asynchronous with the presence of mature females being registered during the entire year. Reproductive aggregations were found in the shallow stratus from late January to August 2009 and reappearing in January 2010. However, the larger ones (which sustained extraction) occurred during June-July and January. An increase in specimen density in the 5 to 17 m stratum was registered in May-August and December-January, coinciding with the periods previous to, during and after the highest aggregation magnitudes registered. The aggregation periods coincided with temperatures over 15°C, with a lower reproductive activity associated to a decrease in temperature. We state that abrupt temperature changes occurring in short time periods could cause mature specimen to move, increasing their density in shallow waters for reproductive purposes. We suggest increasing protection for the resource during the reproductive aggregation processes given that extraction leaves the clutches unprotected and prone to predation from other organisms.
... Considerando que los procesos reproductivos son mejor representados por la gametogénesis de las hembras (), los ejemplares extraídos fueron disectados en el laboratorio y secciones de la zona media del complejo gónada glándula hepática, del 100% de éstas se fijaron en Bouin y se incluyeron en parafina. Cortes de 6 µm de espesor se colorearon con hematoxilina eosina (Gabe, 1968), para su análisis microscópico y se clasificaron según la siguiente escala: Inmaduras: gónada que presenta un predominio de ovocitos que poseen un núcleo central con finos gránulos de cromatina, su citoplasma es basófilo y no se distingue ningún tipo de inclusiones vitelinas. Vitelogénesis inicial: gónada con predominio de ovocitos adheridos a las paredes del folículo que tienen el núcleo con un nucléolo, y en el citoplasma se distinguen sectores hialinos y finas granulaciones eosinófilas correspondientes a gránulos de vitelo. ...
Reproductive and population parameters of Thais chocolata that would contribute to the validation of the current extraction standards were obtained by performing a study in the protected area of La Rinconada Marine Reserve, Antofagasta, Chile, from December 2008 to January 2010. In terms of reproduction, the results revealed asynchronic gonad development in the population, with specimens in different stages of maturity throughout the year. Mature individuals were distributed between 5 and 13 m depth, forming aggregations at 5 m depth during much of the year. Periods of greater maturity were observed from July to August and November to January, with the most important aggregations at the end of both periods. A relationship was observed between the months with the greatest maturity and aggregations and the months with the highest daily temperature fluctuations. At the population level, the results allowed us to estimate 2.3*10(6) individuals, with 39% of this population over the minimum legal size (55 mm). The relatively slow growth estimated for this population was probably influenced by the high variability of the bottom temperature in this area. The critical size and size at first maturity of the population were higher than the minimal legal size, making it is necessary to review the current fishing regulation for this species. The effectiveness of the State marine reserves in the conservation of this marine resource was demonstrated.
... We analysed the morphology of the arms, paying a particular attention to the luminous areas, i.e. the spines. For light microscopy (LM), arm pieces were fixed in 4% paraformaldehyde in filtered sea water, decalcified in a saturated solution of EDTA di-sodium salt in distilled water (adjusted to pH 7 with potassium hydroxide), dehydrated in graded ethanol and embedded in paraffin using a routine method (Gabe, 1968). Sections, 5 µm in thickness, were cut using a Microm HM340E microtome, mounted on clean glass slides and stained with Heidenhain's Azan trichrome (Clark, 1981). ...
Amphiura filiformis is a common burrowing brittle star from muddy environments along the European shores. This species emits a blue light at the level of the arm spine tips when mechanically stimulated. Although the putative luciferase of A. filiformis was recently discovered, the ultrastructure of the luminous cells, i.e. the photocytes, remain unknown for this species as well as for most other bioluminescent echinoderms. In this study, we describe the morphology of the arm and spines in the brittle star A. filiformis and compare it to the luciferase expression pattern revealed by immunohistochemistry. Luciferase is expressed at the base of the spine and at the arm tips within well-defined photocyte clusters. Additionally, we investigated the ultrastructure of spine tissues before and after chemical stimulation of light emission in order to identify photocytes and improve our comprehension of the photogenesis phenomenon in brittle stars. Type II granular cells, found in the basal to mid-basal portion of the spine inner tissues and showing ultrastructural modifications during light-emission, are proposed as photocytes. Putative photocytes appear intimately associated with the spine nerve plexus as well as with mucus cells and presumptive pigment cells. Photocytes are characterised by the presence of specific vesicles with reticulated contents in their cytoplasm. Furthermore, the structure of the calcified spine ossicle has been investigated using microCT scanning.
... Six samples were extracted from each preselected ovary for histological analysis: one from the anterior, middle and posterior sections of the left and right ovarian lobes. These were fixed in Bouin's solution for four to five days, embedded in paraplast, sectioned at a 6 µm thickness (4 to five sections per sampled region), and stained with Gabe and Martoja's one-step trichrome stain (Gabe, 1968). Based on the histological analyses, the preselected females were classified by their gonad development phase into one of the following reproductive phases or sub-phases (Brown-Peterson, Wyansky, Saborido-Rey, Macewicz, & Lowerre-Barbieri, 2011): early developing (ED), developing (D), spawning capable (SC) and actively spawning (AS). ...
Studies of fish reproductive biology often involve quantitative estimates based on histological sections of the ovaries. Prior characterization of gonad oocyte distribution patterns in females is fundamental to these studies. Heterogeneous distribution requires a standardized sampling protocol to prevent bias in data such as fecundity estimations. In contrast, homogeneous distribution allows sampling of any portion of the gonads with no risk of bias. A preliminary study was done of gonad oocyte distribution mode in a hogfish Lachnolaimus maximus population in the southern Gulf of Mexico. A total of 23 sexually-active females were selected and classified by reproductive stage. Six histological sections were taken from the anterior, middle and posterior regions of the left and right ovary lobes of each individual. Digital image processing (AxioVision and Image ProPlus programs) was used to estimate oocyte density per unit area in different development stages. Contingency tables were used to analyze oocyte distribution frequencies between the regions of each lobe and between the lobes of each ovary. This was supported with a Pearson’s χ2 test for goodness-of-fit and a replicated G test to confirm distribution heterogeneity. Oocyte type distribution was homogeneous in almost all the females regardless of ovary region or lobe. This was particularly the case for females in the actively spawning reproductive subphase. Females in this subphase are normally used to estimate batch fecundity in fish species such as L. maximus, which have asynchronous oocyte development and batch spawning. However, heterogeneity was present in tertiary vitellogenesis oocytes in the right lobe of females in the spawning capable reproductive phase. In these cases, it is preferable to take histological sections from any region of the left ovary lobe to ensure unbiased estimation of fecundity and adequate characterization of this species’ sexual cycle.
This study aimed to determine the effect of gallic acid (GA) on ameliorating bisphenol A (BPA) nephrotoxicity in male rat kidneys. Forty rats were assigned randomly into two groups: control (ten animals) and BPA (40 mg/kg bwt) (thirty animals), the second group was divided into three subgroups: BPA alone, BPA + G50 (50 mg/kg bwt), and BPA + G200 (200 mg/kg bwt). The biochemical analysis included measurements of the contents of nitric oxide, lipid peroxidation, reactive oxygen species, and cytokines (interleukin-1α and interleukin-6) in the kidney. The antioxidant enzymes catalase and superoxide dismutase were also measured in the kidney. Kidney function was assessed by determining uric acid, urea, and creatinine levels. The morphological investigations included hematoxylin and eosin staining for assessing the general histology and determining the glomerular and corpuscular areas, the tubular cell degeneration mean area, and the mean leukocyte infiltration area. Also, collagen fiber intensity and polysaccharide content were analyzed. Furthermore, immunohistochemical, morphometric, and ultrastructural studies were carried out. The results revealed morphological, immunohistochemical, and biochemical alterations in the kidney. Most of these changes showed a satisfactory improvement of kidney damage when BPA-administered rats were treated with GA at both doses. In conclusion, GA exhibited a strong protective effect against BPA-induced nephrotoxicity.
Background: Obesity is a significant public health problem that is characterized by an increase in oxidative stress and enhanced inflammatory responses associated with immune cell invasion of adipose tissues. This study assessed several biochemical abnormalities, apoptosis, oxidative stress status, and associated histological changes in the liver, duodenum, and heart brought on by high-fat diet-induced obesity in rats. It also assessed the mechanistic benefits of curcumin in reversing these inflammatory, metabolic, and histological impairments. Methods: Rats were assigned into three groups each including ten rats: the control group (CD), the high-fat diet group (HFD), and the high-fat diet + curcumin (HFDC) group. Serum glucose, insulin, and triglycerides (TAGs) were observed. In addition, apoptosis (indicated by hepatic DNA fragmentation) and oxidative stress status (indicated by hepatic MPO, GSH, and SOD) were assessed. Histopathological examinations included the GIT (liver and duodenum) and heart in addition to quantitative real-time polymerase chain reaction (qRT-PCR) assays of the adipose tissue genetic expressions for inflammatory signaling pathways (TLR4, IL-6, and TNF-α). Results: The overall findings showed that the HFD group exhibited significantly higher levels of glucose, TAGs, and insulin than the control group (P < 0.01). The histological abnormalities of the studied organs in the HFD group were paralleled by these biochemical abnormalities, which were strongly associated with increased apoptosis, increased oxidative stress, and increased expression of the inflammatory signaling markers. There were significant improvements in the HFDC group in terms of biochemical, inflammatory, and histological investigations. Conclusions: This study's findings concluded that obesity is significantly associated with biochemical and microscopic alterations in many organs. Curcumin exerted potent antitoxic, antioxidant, tissue-protective, and antiobesity effects. Curcumin is recommended to be added to various dietary regimens to prevent or delay the organs' dysfunction among obese people.
Standardization of seed production of the Picnic seabream, Acanthopagrus berda, was taken up in captivity. Broodstock development of A. berda was done in 6 m diameter galvanized iron cages during September 2020 to March 2021 using commercial feed (40% protein) supplemented with vitamins and minerals. Males with oozing milt were available consistently throughout the period while asynchronized maturation of female was observed. A pair of mature male and a female (oocyte diameter of 440 ± 12 μm) were induced using a dose of (0.25 ml kg−1) gonadotropin releasing hormone analogue and female was given a similar second dose after 6 h. Spawning occurred after 42 h of induction. Eggs were incubated in fibre glass tanks (100 L) in seawater (34 ppt) with gentle aeration. Number of eggs spawned was 0. 25 million with a fertilization rate of 88% and hatching rate of 80%. Planktonic larvae measured total length (TL) 1.7 ± 0.14 mm at 0 day post‐hatch (dph), started exogenous feeding from 3 dph (TL 2.7 ± 0.48 mm) when the mouth size was 180 ± 2.8 μm. Yolk sac was completely exhausted by 3 dph. Co‐feeding of rotifer Brachionus rotundiformis at 5 numbers (nos) ml−1 and nauplii of copepod (Parvocalanus crassirostris) at 3 nos ml−1 yielded better initial larval survival. Metamorphosis of the larvae started at 25 dph (TL12 ± 0.68 mm) and juveniles emerged from 35 dph (TL 22 ± 0.76 mm). This is the first holistic report about seed production of A. berda.
The paper describes the new land planarian genus Mapuplana gen. nov. from Chile, on the basis of the two new species Mapuplana guttulata sp. nov. and M. fjordica sp. nov. The genus Mapuplana is mainly characterised by two putative apomorphies: a subneural parenchymal musculature consisting of diagonal decussate muscle fibres; and a blind duct opening sideways into the female atrium. The new species are very similar to each other in their general anatomy and differ only in details in the pattern of dorsal colouration, the relative distance between mouth and gonopore, the relative thickness of the ventral cutaneous musculature, the orientation of the gonoduct, and the shape of the female atrium. http://www.zoobank.org/urn:lsid:zoobank.org:pub:120AC180-D4AC-433F-B3A0-BB96C6962DB4
This article describes the morpho‐anatomies of the leaves and stems of Ocotea paranaensis Brotto, Baitello, Cervi & Santos (Lauraceae) using light and scanning electron microscopy. The main anatomical features characterizing the species are the presence of simple non‐glandular trichomes in the leaves and stems, large secretory cells in the leaves, flat‐convex petioles with two small lateral ribs, brachysclereids in the stem cortex and pith, and the presence of starch grains and various types of calcium oxalate crystals in the pith. Histochemical tests indicated the presence of lignin in stone cells, fibers, and xylem. Lipophilic contents were found in the secretory cells. Phenolic compounds were detected in the epidermis, hypodermis, phloem, and xylem. The present study's findings can contribute to the taxonomy and authentication of O. paranaensis. This article is the first morpho‐anatomical study of Ocotea paranaensis. Anatomy and histochemistry of the leaves and stems were studied by light microscopy, FESEM and EDS. The findings of this study would aid in the species identification and taxonomy. Morpho‐anatomy of leaves of Ocotea paranaensis (b, e: light microscopy, sections stained with safranin; c, d, f, g: FESEM). a: a twig of plant. b–g: leaf epidermis in surface view (b, c: adaxial side; d–g: abaxial) (ct, cuticle; md, midrib; nt, nonglandular trichome; st, stomata; wa, epicuticular wax). Scale bars: a = 2 cm, d, f = 100, b, c, e = 50, g = 20 μm.
Broodstock development, induced maturation and breeding of the newly described seabream, Sparidentex jamalensis collected from Kali estuary, Karnataka, India were achieved successfully for the first time. Taxonomy of the fish was confirmed employing morphological and molecular tools. Majority of the brooders collected were in the 20–30 cm length class (140–510 g) and were ideal for developing matured broodstock. Periodic microscopic observation of ovarian biopsy indicated progressive gonadal development in the broodstock. The broodstock developed for this protandrous hermaphrodite consisted of females having maturing oocytes and males with oozing milt during the breeding season. Final maturation and spawning of the females (520 ± 8 g; ova diameter above 400 µm) was achieved using cholesterol‐based LHRH analogue pellets (50 µg/kg fish) and the males (450 ± 54 g) were not treated with hormones. Pelagic fertilized eggs (720 ± 8 µm) with a single oil globule were obtained with an absolute fecundity of 0.42 ± 0.06 million/kg fish and a fertilization rate of 82 ± 2.4%. Major embryological stages were recorded and the fertilized eggs hatched after 18–20 hours with 80 ± 4.6% hatching rate at 28°C. The newly hatched planktonic larvae measured 1.8 ± 0.12 mm in total length. Mouth opening of the larvae was observed on second day post hatch (dph). Resorption of the yolk sac was observed on third dph. These results will act as a baseline for developing a breeding programme for this species.
Oregano essential oil (OEO), one kind of Labiata has been known for a long time as a popular remedy. The essential oil of oregano has anti-bacterial, anti-oxidant, anti-fungal and nematicidal properties mainly due to the basic constituents. Once the shell’s microbial barriers have been breached, Gram-negative bacteria are more capable of withstanding the antimicrobials present in the albumen. Eggs were divided into, control group (GI), LPS group (GII) and LPS plus OEO 0.5% group (GIII), as natural disinfectants. Statistical data were analysed, histopathological, RT-PCR and in situ hybridization studies were performed to explain the OEO modulatory role. Lipopolysaccharide (LPS) group represented the highest (P < 0.05) increase of embryonic mortality percentage, greatest significance (P < 0.01) egg weight loss, a decrease of hatchability and high teratogenic effect compared with those for other groups in all types of experiments. Chicks produced from egg subjected to oregano 0.5% and LPS significantly (P < 0.05) recorded large body weight gain, enhancement of hatchability and produces no teratogenic effects in the developing chicken. OEO modulates the effect of LPS in histopathological, RT-PCR and in situ hybridization studies throughout modulation of the Daam2 and Tbx2 gene expression towards the normal one. Using Oregano oil as a natural disinfectant in this study modulate significantly the effect of LPS oxidative effect on the chick embryo even after hatching, so, we conclude that OEO is an effective anti-oxidant that recommended to be used in hatchery for good quality chicks, chick growth and finally improvement of laying production performance.
This study evaluated the toxicity of citric acid and the benefits of soya milk (SM) for preventing damage in mice. Thirty-five mice were divided into groups: control, mice administered citric acid (CA group) for 30 days, mice administered SM before the administration of citric acid for 30 days (SM + CA group), mice administered citric acid for 15 days and left for recovery (R group), and mice in recovery receiving SM for 15 days (R + SM). Mice in CA and R groups displayed downregulated p53, increased cleavage of caspase 3, and upregulation of Nrf2, CYP1A1, ALT, and AST activity in the liver. In contrast, SM + CA and R + SM treated mice were protected against CA toxicity and showed reversal of p53 downregulation, reduced cleavage of caspase 3, downregulation of Nrf2, and an increase in liver function enzymes. SM administration also restored blood cell and hemoglobin content and general histology of hepatocytes. Practical applications CA causes liver damage, increases inflammation, decreases blood cell numbers, and induces apoptosis. Some natural products, such as SM, have been used to scavenge free radicals that can cause liver damage and hemolysis. This study focuses on the effectiveness of SM in ameliorating CA toxicity and may be helpful in the food industry for managing oxidative stress that may be induced by common dietary constituents. SM may help suppress liver damage and inflammation.
Studying the dermal skeleton in fish is valuable for phylogenetic specification. The current study describes the detailed structure of the plecostomus dermal skeleton, including its morphogenesis and distribution in the skin. The denticles have a crown and a basal part and are embedded in bony depressions, to which they are attached by denticle ligaments. During denticle morphogenesis, denticle papillae formed from denticle precursor cells align in two cellular layers: an outer ameloblast precursor layer and an inner odontoblast precursor layer. The ameloblast precursors and odontoblast precursors differentiate and secrete enamel and dentine, respectively. We used different histochemical techniques, including Crossmon's trichrome staining, Weigert–Van Gieson staining, periodic acid–Schiff (PAS) staining, combined Alcian blue (AB; pH 2.5)/PAS staining, Weigert–Van Gieson staining, Mallory trichrome staining, and AB staining to distinguish the dentine and denticle ligaments. We used acridine orange to detect lysosome activity during denticle eruption. Transmission electron microscopy was used to detect the denticle ultrastructure, and scanning electron microscopy was used to detect the topographic distributions of different types of dermal tissues in different anatomical regions.
An experiment was conducted for 120 days to evaluate optimum dietary lipid requirements for gonadal maturation of Cyprinus carpio fed with varying dietary lipid levels under biofloc‐based systems (BFT). About 180 fingerlings (22 g ± 0.05) were randomly distributed in 15 tanks (300 L) at the rate of 48 no./m2 and fed with varying lipid levels (T1—4% lipid with BFT, T2—6% lipid with BFT, T3—8% lipid with BFT, T4—10% lipid with BFT, control—10% lipid without BFT). The C/N ratio of 20:1 was maintained using tapioca flour as carbon source. Biofloc units supplemented with 8% dietary lipid (T3) showed advanced maturation in terms of absolute fecundity (9,913 ± 7.62), relative fecundity (229.0/g ± 11.92), gonadosomatic index (24.47% ± 1.27), hepatosomatic index (1.97% ± 0.07), condition factor (0.02 g/cm3 ± 0.00) compared with control (clear water with 10% lipid) (p < .05). Histological observations of gonads also revealed that the biofloc groups with supplementation of 8% dietary lipid promoted gonadal maturation for female oocyte and 6% dietary lipid promoted maturation for males, compared with control (clear water with 10% lipid). The results obtained in this experiment elucidate that the biofloc improves gonadal maturation of common carp broodstock at a dietary supplementation with 8% lipid compared with conventional system of broodstock management.
The histological study of hard pieces such as tendons and calcified lesions and tissues is a field that has been gaining increased attention owing to the rapid development of implantable prostheses, among other factors. In these studies, serial sectioning is utilized to detect areas of interest throughout the entire piece, as it enables the application of the appropriate light and electron microscopy techniques in these areas. We propose the "three-sectioning method" that subjects the pieces to three consecutive cycles of embedding and sectioning to localize and study the areas of interest, as an efficient technique for these histological studies. The pieces were first embedded in epoxy resin and then cut into thick sections (approximately 300 μm) for the first cycle. Next, areas of interest selected on these thick sections were re-embedded in epoxy resin to be sectioned again (second sectioning) to obtain a series of semithin sections (1-3 μm). These semithin sections are usually studied using the most relevant techniques for light microscopy. Smaller areas of interest are selected to be cut into ultrathin sections (60-90 nm) for transmission electron microscopy. If necessary, the selected areas of the semithin sections can be embedded again, and then cut into new ultrathin sections. The different kinds of sections we have described here may also be studied using scanning electron microscopy. This systematic method facilitates correlative microscopy from lower to higher magnifications along with the usage of a broad variety of histological techniques including electron microscopy.
Micromorphological studies were carried out using multiple microscopic techniques on the leaves and stem bark of Byrsonima sericea DC. (Malpighiaceae), a species popularly known as “murici” and used medicinally, in order to identify both qualitative and quantitative features of leaf and stem anatomy and histochemistry as differential parameters to support both the quality control of its ethnodrugs and the taxonomy of the genus. The study was conducted using traditional techniques of plant anatomy, histochemical tests, and the stomatal index (SI). Byrsonima sericea has hypostomatic leaves, anomocytic stomata, and its epidermal walls are anticlinal and straight on the adaxial and curved on the abaxial faces. T‐shaped trichomes were observed mainly on the abaxial surface. The leaf epidermis showed waxes syntopism on both surfaces, with the occurrence of different crystalloid forms on a single phylloplane. The mesophyll is dorsiventral, with 3‐4 collateral vascular bundles. Phenolic compounds, starch, and proteins were identified in the petiole and stem. The SI was 14.5 ± 0.53% (p < .05), but did not showed significant variations. A set of characters were found to be distinctive for the studied species, however, constituting parameters that could be used to separate B. sericea from other species of the genus. The abaxial surface is ferruginous, with T‐shaped trichomes. The leaf epidermis shows syntopism of epicuticular waxes, which, together with the shape of the vascular bundles can be used to separate B. sericea from other species of the genus.
The annual reproductive cycle of picnic seabream, Acanthopagrus berda (Forsskål, 1775), one of the potential aquaculture candidate from estuarine waters of Calicut, Kerala (India) was studied. Based on the morphological and histological studies, the ovotestes of A. berda were classified as active male, active female, inactive male, inactive female, transitional and undifferentiated. Histological observation of transitional gonads showed signs of degeneration in the testicular lobe, proliferation of connective tissue and empty sperm ducts indicating protandrous hermaphroditism in A. berda. Ovary was classified into seven maturity stages (virgin, developing virgin, developing, maturing, mature, running, spent) and testis into five maturity stages (resting, maturing, mature, running, spent). Gonadal development in A. berda indicated resting phase (February–July), pre‐spawning phase (March–August) and spawning phase (August–December). Inactive (24.6%) and active males (21.6%) were observed as dominant in smaller length classes (140–250 mm TL), whereas inactive (18%) and active females (51%) were observed as dominant in larger length classes (251–450 mm TL). Few primary females (28.1%) were observed in smaller (below 250 mm TL) and few primary males (28.5%) were observed in larger length classes (above 250 mm TL). From the present study, it can be concluded that in A. berda, most of the individuals function first as males and then change sex to female, but few continue to function as either male or female throughout their lifespan indicating digynous protandrous hermaphroditism.
Premise: Laticifers have evolved multiple times in angiosperms and have been interpreted as a key innovation involved in plant defense mechanisms. In Malpighiaceae, laticifers were previously known from a single lineage of trees and shrubs, the Galphimia clade, but with detailed anatomical analyses here, we show that their distribution is broader in the family, also encompassing large clades of lianas. Methods: From 15 genera, 70 species of Malpighiaceae were surveyed through careful anatomical ontogenetic analysis of roots, stems, and leaves and detailed histochemical tests to elucidate the nature of laticifers and latex in the family. Results: Articulated anastomosing laticifers were encountered in roots, stems, and leaves of two distantly related megadiverse genera of Malpighiaceae lianas: Stigmaphyllon (stigmaphylloid clade) and Tetrapterys s.s. (tetrapteroid clade). From the apex downward, in Stigmaphyllon the laticifers are derived from the procambium and from the cambium during its early activity and are present in the outermost part of the vascular cylinder of stems and leaves and in the pericycle of roots, whereas in Tetrapterys s.s. they are derived from the ground meristem, procambium, and cambium throughout the plant body and are present in the cortex and pith, either the pericycle in roots or the outermost part of the vascular system in stems and leaves, and the primary and secondary phloem. Conclusions: Laticifers seem to have evolved at least three times independently in Malpighiaceae, once in a lineage of trees and shrubs and twice in two distantly related megadiverse lianescent lineages. Laticifer evolution in Malpighiaceae is homoplastic and may be related to increases in species diversification.
Light microscopy studies of the female American lobster Homarus americanus reproductive system are essentially nonexistent or outdated. Based on samples taken in the spring, summer, and autumn from the southern Gulf of St. Lawrence between 1994 and 2014, and using a combination of histological and scanning electron microscope techniques, we propose an ovarian cycle with 10 stages, identifying for the first time a recovery stage. Also, an atypical resorption stage, characterized by massive reabsorption of mature oocytes, is occasionally observed during summer months. The oviducts are composed of connective tissue (elastic and collagen fibers) with no muscle or secretory activities. Their epithelium shows a cyclic pattern and phagocytosis activities linked to spawning. Although the role of the seminal receptacle is to store and protect semen, free spermatozoa (i.e., without the spermatophoric wall and the acellular gelatinous substance that constitute the semen) were also observed in its posteriolateral grooves immediately prior to spawning, which is consistent with an external fertilization mechanism at the seminal receptacle. Unexpectedly, free spermatozoa were observed externally near two pore‐like structures located on the gonopore's operculum, not at the seminal receptacle, after spawning; hence, more work is needed to fully understand the fertilization mechanism for the American lobster.
Despite supporting a valuable fishery, the reproductive system of the male American lobster (Homarus americanus) is poorly understood. The elongated H‐shaped testis is responsible for spermatogenesis and is composed of follicles, a common collecting duct with interlaced scattered striated muscles, and a serosa as an external wall. Sertoli cells are associated with the spermatogenesis that produces spermatozoa, which are transferred to the collecting duct through a temporary passageway. Spermatogenesis is asynchronous between follicles and occurs on a continuous basis. The anterior and posterior lobes of the testes are independent and connect to the vasa deferentia through the Y‐shaped collecting tubules that have a different cell anatomy and function than the two organs they connect. The vas deferens is divided into four regions. Spermatophores, produced in the proximal vas deferens, are packets of spermatozoa encapsulated in a single layer—the spermatophoric wall, which is composed of mucopolysaccharide acid. Large dense ovoid granules and the seminal fluid, composed of acidic sulfated mucosubstances, are secreted in the median vas deferens. Spermatophores within these secreted substances (i.e., semen) are stored in the distal vas deferens that, with the spermiduct (last region of the vas deferens), is responsible for the extrusion of the semen by striated muscle contractions. Smooth muscles suggest a peristaltic movement of the spermatophores within the vas deferens. Finally, the gonopores and the first pair of pleopods (i.e., gonopod) move the semen to the female seminal receptacle during copulation.
The chapter provides a concise account of collecting techniques, and basic histological techniques for investigating planarians, intended for use by practicing scientists and postgraduate students new to the field. Selected hands-on methods are described that will lead to well-stained preparations suitable for light microscopic examination for subsequent taxonomic identification or general investigations into planarian functional cytology and histology. Major equipment items that are minimal essential requirements of a functional histological laboratory are indicated. The chapter includes sectioning and staining troubleshooting guides that will be very helpful for those who must work in the laboratory without the opportunity to consult qualified and experienced technicians or microscopists.
Hundreds of planarian species exist worldwide, representing a rich phenotypic diversity. This chapter presents an overview of the morphology and anatomy of various taxonomic groups of planarian flatworms, focusing on features enabling recognition and identification of the animals. The most recent view on the phylogenetic relationships of the planarians is presented, together with geographic distribution patterns of major groups of triclads. The chapter concludes with a brief methodological section outlining species identification on basis of anatomical features. In conjunction with the established laboratory model species, the phenotypic diversity of planarians provides rich opportunities for comparative studies, which this chapter aims to inspire.
This study reports a detailed anatomical and histological study of the digestive system of Octopus vulgaris. Emphasis was placed on characterising the glands and glandular cells and their distribution throughout the digestive tract. The use of classic histological and histochemical techniques revealed two morphological types of glandular cells: granular and mucous. Moreover, the histochemical analysis indicated specialisation of mucous glandular cells in the buccal mass, the submandibular gland and the caecum for secreting acid and neutral glycoconjugates. The cells of the anterior salivary glands are specialised for secreting neutral glycoproteins, and those of the posterior salivary glands are specialised for granular and mucous secretion. The oesophagus, crop and stomach lack glandular cells, but both granular and mucous glandular cells are found in the intestine. An unusual structure resembling the typhlosole of bivalves is described for the first time in the intestine of O. vulgaris. The highly ciliated epithelium and location of the structure in the anterior part of the intestine suggest a possible role in bypassing the caecum, stomach and intestine. We discuss how these cells and organs contribute to the process of digestion in the light of the present histological and histochemical data and of previously published information on the morphology and physiology of digestion in the octopus.
The Citrullus colocynthis, commonly called colocynth, is known because of its purgative effects and whose seeds are commonly used as certain diseases treatment, namely liver diseases, in the Mediterranean countries traditional medicine. This study aims to analyze the effect of two colocynth extracts « glycosides » and « alkaloids » on metabolic and histological disorders associated with liver function in Wistar rats (Rattus norvegicus). This pathology is due to an enriched oil palm diet. For this purpose, Wistar male rats n = 18, weighing between 130g and 150g, are divided into two lots. A control group (C) n = 6, receives a standard laboratory diet ; an experimental group (E) n = 12, receives a standard laboratory diet supplemented with palm oil. After seven months of experimentation, 8 experimental rats were sacrificed for the morphological study and the remaining 12 rats undergo a colocynth treatment (Tr) for eight weeks. They are subdivided into: The first six experimental rats receive a 70mg/kg single intraperitoneal injection of ethanol extract of cucurbitacin glycosides (Glc). The second lot receives a 70mg/kg single intraperitoneal injection of total alkaloids extract (Alc). The animals of (E) group showed hyperglycemia, hyperinsulinemia, hyperlipemia, dyslipoproteinemia, a significant increase of the enzymatic activity of transaminase (AST and ALT) and alkaline phosphatase (ALP). Histological examination of the liver gland shows major damages Non-alcoholic steatohepatitis [NASH]. Treatment with colocynth glycosides and alkaloids reveals a significant improvement at different levels in plasma as well as in tissue. Treatment with colocynth glycosides and alkaloids shows a hypoglycemic effect, lipid-lowering a well as a hepato-protective effect.
Titanium dioxide nanoparticles (TiO2NPs) are used extremely and represent one of the top five most regularly used nanoparticles worldwide. Currently, numerous studies referred to their toxic possible on organs using different treatment course. Materials and methods: The present study was carried out to evaluate the potential protective role of blackberry and selenium (Se) against (TiO2NPs)-induced oxidative stress and histological changes in liver and heart tissues of rats. Rats were divided into seven groups according to the treatment into control, Titanium dioxide treated group (TiO2NPs) (500 mg/kg B.W.), Black berry (Bb) treated group (1.6 g/kg body), Selenium (Se) (0.5mg/kg;), (TiO2NPs)plus Bb and (TiO2NPs)plus Se and the last group treated with combination of (TiO2NPs) plus Bb and Se. Results: All the Animals were treated orally for 30 successive days. (TiO2NPs) increased alkaline phosphatase (ALP), aspartate aminotransferase (AST), alanine aminotransferase (ALT), lactate dehydrogenase (LDH) and γ-glutamyl transpeptidase (γ-GT) activities and changed the levels of lipid profile as well as creatinine and uric acid levels. It marked decreased hepatic reduced glutathione (GSH) levels, antioxidant enzymes activities and increased the levels of lipid peroxidation. Bb and Se prevented the (TiO2NPs)-induced liver injury as indicated by improving all the parameters previously illustrated. Histopathological results confirm the biochemical finding and the ameliorating effect of both Bb and Se on liver toxicities and heart failure. Conclusion: Cotreatment of either Bb or Se possessed different protective mechanisms against (TiO2NPs)- induced liver toxicity and heart damage. So, the intake of (TiO2NPs) should be restricted due to it's increased toxicity.
In this paper, we focus on the reproductive behavior of the cockroach Oxyhaloa deusta. Within the Oxyhaloinae subfamily, mating strategies and glandular areas involved in pheromone production have been studied for several genera belonging to the Nauphoetini and Gromphadorhini tribes. However, this is the first time that courtship and mating behavior have been explored within the genus Oxyhaloa, and in a more general way within the Oxyhaloini tribe. This work, comprising behavioral observations, olfactometric bio-assays and morphological data, highlights unusual behaviors and novelties in potential sex pheromone gland location, in both males (tergite 8) and females (supra-valvular area). Surprisingly, our results also indicate that mate finding is initiated by the female. This is quite remarkable inasmuch as the Oxyhaloinae subfamily is the only cockroach group in which males initiate mate finding by means of a sex pheromone, emitted during the calling posture by extending abdominal tergites. In the Oxyhaloinae subfamily, this occurrence of various reproductive behavioral patterns (including all the mating patterns known at present in Blattaria) in closely-related species is striking, and makes this group a suitable model for studying the changes in mating behavior correlated with the location of sexual glands.
The Laurentian Great Lakes contain one fifth of the world’s surface freshwater and have been impacted by human activity since the Industrial Revolution. In addition to legacy contaminants, nitrification and invasive species, this aquatic ecosystem is also the recipient of Contaminants of Emerging Concern (CECs) with poorly understood biological consequences. In the current study, we documented the presence, concentrations, and biological effects of CECs across 27 field sites in six Great Lakes tributaries by examining over 2250 resident and caged sunfish (Lepomis ssp.) for a variety of morphological and physiological endpoints and related these results to CEC occurrence. CEC were ubiquitous across studies sites and their presence and concentrations in water and sediment were highest in effluent dominated rivers and downstream of municipal wastewater treatment plant discharges. However, even putative upstream reference sites were not free of CEC presence and fish at these sites exhibited biological effects consistent with CEC exposure. Only the Fox River exhibited consistent adverse biological effects, including increased relative liver size, greater prominence of hepatocyte vacuoles and increased plasma glucose concentrations. Canonical Redundancy Analysis revealed consistent patterns of biological consequences of CEC exposure across all six tributaries. Increasing plasma glucose concentrations, likely as a result of pollutant-induced metabolic stress, were associated with increased relative liver size and greater prominence of hepatocyte vacuoles. These indicators of pollutant exposure were inversely correlated with indicators of reproductive potential including smaller gonad size and less mature gametes. The current study highlights the need for greater integration of chemical and biological studies and suggests that CECs in the Laurentian Great Lakes Basin may adversely affect the reproductive potential of exposed fish populations.
The current study aimed at evaluating the ability of a mineral and antioxidant-rich extract from Chondrus canaliculatus to improve maneb (MB)-induced toxicity in adult rat. The animals were divided into four groups: group 1 used as a control group, group 2 received MB, group 3 received MB + C. canaliculatus extract, and group 4 received only the algal extract. MB, a Mn-containing ethylene-bis-dithiocarbamate fungicide, induced oxidative stress damages, mineral perturbations in the plasma, urine, and bone, and genotoxicity in rats. Hematological analysis revealed in the MB-treated group a disruption in the number of red blood cells, platelets, and white blood cells associated with a striking genotoxicity. Interestingly, a significant increase in malondialdehyde and advanced oxidation protein product levels in erythrocytes and bones were found. On the other hand, an impairment of the antioxidant status in both tissues was occurred. Along, our results revealed that MB injection caused a striking drop and disruption in bone’s mineral rates, especially calcium and phosphorus. These biochemical results were in accordance with the histological and molecular changes. However, co-treatment with C. canaliculatus extract showed, for the first time, that this alga was effective against MB-induced hematotoxicity, genotoxicity, and oxidative stress in the blood and bone and maintained osteomineral metabolism and bone histo-architecture. Such observations might be explained by the strong in vitro antioxidant and antibacterial activities exhibited by the alga, as well as by its high levels in several minerals: calcium, phosphorus, sodium, potassium, magnesium, iron, and zinc.
Squamates present a unique challenge to our understanding of dental evolution in amniotes because they are the only extant tooth-bearing group for which a ligamentous tooth attachment is considered to be absent. This has led to the assumption that mammals and crocodilians have convergently evolved a ligamentous tooth attachment, composed of root cementum, periodontal ligament, and alveolar bone, whereas squamates are thought to possess a single bone of attachment tissue that fuses teeth to the jaws. The identity and homology of tooth attachment tissues between squamates, crocodilians, and mammals have thus been a focal point of debate for decades. We provide a novel interpretation of the mineralized attachment tissues in two focal taxa in this debate, mosasaurids and snakes, and compare dental tissue histology with that of the extant crocodilian Caiman sclerops. We identify a periodontal ligament in these squamates that usually exists temporarily as a soft connective tissue anchoring each tooth to the alveolar bone. We also identify two instances where complete calcification of the periodontal ligament does not occur: in a durophagous mosasaur, and in the hinged teeth of fossil and modern snakes. We propose that the periodontal ligament rapidly calcifies in the majority of mosasaurids and snakes, ankylosing the tooth to the jaw. This gives the appearance of a single, bone-like tissue fusing the tooth to the jaw in ankylosed teeth, but is simply the end stage of dental tissue ontogeny in most snakes and mosasaurids.
The ultrastructure of the ovary during development and yolk production is poorly known in Brachyura and Majoidea in particular. Here, we describe the histology, histochemistry and ultrastructure of the adult ovarian cycle in four Mithracidae species from three different genera: Mithrax hispidus, Mithrax tortugae, Mithraculus forceps and Omalacantha bicornuta. All species showed a similar pattern of ovarian development and vitellogenesis. Macroscopically, we detected three stages of ovarian development: rudimentary (RUD), developing (DE) and mature (MAT); however, in histological and ultrastructural analyses, we identified four stages of development. The oocytes of the RUD stage, during endogenous vitellogenesis, have basophilic cytoplasm filled with dilated rough endoplasmic reticulum. The reticulum lumen showed many granular to electron-dense materials among the different stages of development. The Golgi complexes were only observed in the RUD stage and are responsible for releasing vesicles that merge to the endogenous or immature yolk vesicles. At the early DE stage, the oolemma showed many coated and endocytic vesicles at the cortex. The endocytic vesicles merge with the endogenous yolk to form the exogenous or mature yolk vesicles, always surrounded by a membrane, characterizing exogenous vitellogenesis. The exogenous yolk vesicles comprise glycoproteins, showing only neutral polysaccharides. At the late DE stage, endocytosis still occurs, but the amount of endogenous yolk decreases while the exogenous yolk increases. The late DE stage is characterized by the beginning of chorion production among the microvilli. The MAT stage is similar to the late DE, but the endogenous yolk is restricted to a few cytoplasmic areas, the ooplasma is filled with exogenous yolk, and the oolemma has very few coated vesicles. In the MAT stage, the chorion is fully formed and shows two electron-dense layers. The ovarian development of the species studied has many similarities with the very little known Majoidea in terms of the composition, arrangement and increment of the yolk vesicles during oocyte maturation. The main differences are in the vitellogenesis process, where immature yolk formation occurs without the direct participation of the mitochondria but with the participation of the rough endoplasmic reticulum in the endogenous phase.
Male albino rats of Wistar strain were injected intraperitoneally with nicotine or/and resveratrol for 4 weeks. Serum Interleukin-2, Interleukin-6, alpha-fetoprotein and tumor necrosis-alpha, as well as plasma 8-hydroxydeoxyguanosine of nicotine-treated rats were increased significantly. Myeloperoxidase, xanthine oxidase, nitric oxide, lipid peroxidation and total oxidative status of the lung in nicotine-treated rats were increased significantly, which were brought down to normal in resveratrol co-treated group. Endogenous antioxidant status as the activity of superoxide dismutase, catalase, glutathione peroxidase, and glucose-6-phosphate dehydrogenases were found to be decreased significantly in the lung of the nicotine-treated group, which were significantly raised in resveratrol-administered groups. The non-enzymatic antioxidants as total antioxidant and thiol levels were decreased significantly as the effect of nicotine that was effectively enhanced by resveratrol treatment. The lung of nicotine-treated rats showed severe congestion of the alveolar lung tissues with scattered congestion per bronchiolar and perivascular cells, as well as, inflammatory cells were observed. The data suggested that resveratrol exerts its protective effect by modulating the extent of oxidative status and improving the enzymatic/non-enzymatic antioxidant defense system, moreover, decreases the pathological changes in animals against the lung damage caused by nicotine.
Inserted in one of the largest families among the Angiosperms, Genipa americana L., Rubiaceae, can be found in all Brazilian territory, presenting great medicinal importance, where several uses have been attributed. In view of this, this work has the purpose of analyzing the ultrastructural, biomineral, phytochemical and histochemical characteristics of the leaves of this native species from Brazil. For this, light microscopy, polarization and scanning electron microscopy techniques were used with X-ray scattering energy, associated to chromatographic and histochemical tests. The anatomical ultrastructural characteristics of the leaves detailed information about the type and arrangement of the cuticle, trichomes, surface and arrangement of the tissues that determine the botanical identity of this species. The phytochemical tests allowed determining their chromatographic pattern and histochemistry to determine the exact storage site of these substances in the leaf. It was observed that the characterization of the crystalline macro-pattern present in the analyzed species, as well as its exact elemental composition, can be considered an important differential diagnosis factor. The results characterize the leaves of this species in different aspects, being a native species and pharmacologically promising, with different popular uses and proven pharmacological activities, and more in depth studies is needed.
Many Viburnum species are popular ornamental shrubs and, simultaneously, highly valued medicinal plants as a source of many bioactive compounds, including antioxidants. Viburnum bark, flowers, and fruits are widely used in traditional and folk medicine, and the fruits of some species are used as cooking ingredients. The knowledge of the microstructure of Viburnum fruits and the accumulation sites of bioactive substances in these organs is rather poor. Comparative analyses of the microstructure of ripe Viburnum opulus and Viburnum lantana drupes were carried out using light, scanning, and transmission electron microscopes. The location of various groups of metabolites in the fruits of both species was determined with the use of histochemical tests and fluorescence microscopy. Additionally, the major antioxidants, i.e. carotenoids, polyphenols, and flavonoids, were quantified and a number of morphometric traits of the drupes were presented. The V. opulus and V. lantana fruits were found to differ in some morphological traits and in many characteristics of the pericarp anatomy and ultrastructure. It was shown that the Viburnum fruits contained lipids and lipid compounds (carotenoids, essential oils, steroids, and saponins), polyphenols (tannins, flavonoids, and anthocyanins), pectins, and proteins. The fruits of V. opulus contained greater quantities of carotenoids, polyphenols, flavonoids, steroids, and pectins than the V. lantana drupes, whereas the latter were characterised by higher contents of essential oils, saponins, and proteins. The metabolites were located in different pericarp layers, but the greatest amounts were identified in the drupe skin.
The larval presence of M. chilensis was determined monthly from September 2007 to March 2008, in Quillaipe, Metri, Sotomo, Canutillar, and Pichicolo (interior sea of Chiloé) using vertical plankton trawls through a water column from 0 to 7.5 m. At the same time, the reproductive stage of adult individuals was determined histologically. Larvae were detected continuously at the five study sites, with a significant increase from October to December, exceeding 49 thousand per m-3 at Canutillar where, along with Sotomo, the greatest densities were recorded during the study. Cohort analysis showed a similarity in the larval cohorts present at the different sites from September onwards; this was accentuated in November and December, as was the constant appearance of cohorts in the initial stages. In contrast with these results, the reproductive process of M. chilensis at these sites occurred from January to March. Whether or not the spawning recorded at these sites is responsible for the observed larvae is a matter of debate, as the sites may receive and retain larvae exported from other sites. These results suggest a need to compile further biological and ecological knowledge on the areas where this species is distributed and its larval and post-larval dynamics in order to identify exporting and retention areas and thereby contribute to efficient management and the optimization of spat collection.
The global dynamic spread of chemical contamination through the aquatic environment calls for the development of biomarkers of interest. Reproduction is a key element to be considered because it is related to the sustainability of species. Spermatogenesis is a complex process that leads to the formation of mature germ cells, whose steps and impairments need to be finely described in ecotoxicological analyses. The physiological process has been commonly described by histological analyses of gonads in different taxa. In the present paper, we describe the development of a novel technique to characterize spermatogenesis based on the analysis of the DNA content of germ cells by flow cytometry, using a DNA-intercalating agent. This new biomarker, referred to as an index of sexual maturity, proved relevant to describe the seasonal reproductive cycle of the zebra mussel, Dreissena polymorpha (Pallas, 1771), used as a sentinel species in the biomonitoring of continental waters and sensitive to highlight the reprotoxicity of carbamazepine (an anti-epileptic pharmaceutical) tested under ecosystemic conditions (mesocosms).
When applied to hermaphrodite organisms, the local mate competition hypothesis predicts an increase of the ratio of sperm to ova produced as the number of mates increases. Here we test this prediction using a hermaphroditic platyhelminth parasite (trematode), Echinostoma caproni. This worm inhabits the small intestine of vertebrates, inevitably inducing the formation of highly subdivided populations, a condition known to promote local mate competition. Moreover this echinostome exhibits an unrestricted mating pattern involving both selfing and outcrossing as well as multiple fertilizations. We quantified the investment in reproductive organs by estimations of testes, cirrus sac, ovary, and egg size and fecundity when echinostomes were isolated alone, in pairs, or in groups of 20 worms. Adult body size was also recorded as a covariate. When mating group size increases (singles, pairs, or groups) we observed a significant increase in resource allocation to male function in addition to a significant decrease in ovary size. Smaller ovaries do not seem to affect egg size, but do result in a reduction in fecundity. Finally, our results are in accordance with the expected theoretical relationship between male allocation and the number of potential mates given local mate competition.
The production of the flat oyster Ostrea edulis (L.) natural spat in Europe has decreased almost by 60% in the past ten years. Thus, the importance of the production of oyster spat in hatcheries is evident. One of the critical steps in hatchery production is broodstock conditioning, especially difficult in autumn, when gonadal development is in resting period. Conditioning is influence by temperature, photoperiod and nutrition. In this work, the effects of two temperature and three photoperiod regimes on the conditioning of O. edulis were studied for three years by stereological analyses and registering number and dates of spawning and larval yield. Temperature had a positive effect on the gonadal development of O. edulis during conditioning. The percentages of germinal cells in oysters conditioned with a gradient of temperature (14-18°C) were double compared to oysters conditioned at 15°C. Oysters conditioned with longer photoperiods showed higher percentages of germinal cells. There was no interaction between temperature and photoperiod. Spawning was observed in the oysters treated with daylight (8-16 h) ten weeks from the beginning of conditioning. Flat oysters conditioned with 8 h and 8-12 h of daylight delayed the first spawning for a month. Total larval production was higher in the oysters treated with the longest daylight gradient. Gonadal and gametogenic development was a non-synchronic process and the spawning extended for around two months. A protocol for flat oyster broodstock conditioning in autumn by using both a gradient of temperature (14-18°C) and daylight (8-16 h) is proposed.
Anatomical and ultrastructural investigations of the glandular trichomes of aboveground organs of the white horehound (Marrubium vulgare L.) were carried out. Additionally, identification of the main components of the secreted products was conducted. The following types of glandular structures were found: peltate with a stalk cell and a large eight-celled head and capitate trichomes, which were divided into three types: I. long-stalked with a 1-celled head, II. short-stalked with a 2-celled head, III. short-stalked capitate with a 4-celled head. Histochemical tests showed that the glands contained various substances such as lipids, polysaccharides, phenolic compounds, terpenes, tannins, and flavonoids. During the secretory stage, the ultrastructure of the bicellular head of the capitate trichome was characterized by a highly developed rough endoplasmic reticulum, numerous plastids, and mitochondria. After exocytosis, the secretion was temporarily stored in the periplasmic space and passed through the cell wall to the subcuticular space.
The aim of this study was to elucidate the redox effects of Thiophanate methyl (MT) in the rat liver and kidney. Our results showed, after 3 days of MT injection (700 mg/kg), an increase in malondialdehyde (MDA), hydrogen peroxide and advanced oxidation protein products levels. Glutathione peroxidase and superoxide dismutase activities were also remarkably increased in the liver but decrease in the kidney. Glutathione and vitamin C values were significantly reduced. The changes in biochemical parameters were substantiated by histological and molecular data. A smear without ladder formation on agarose gel was shown, indicating random DNA degradation in the liver and the kidney of MT treated rats. The increase in cyclooxygenase-2 gene expression, marker of inflammation, and an increase in genes expression of glutathione peroxidase and superoxide dismutase in liver and their decrease in the kidney were also occurred after MT exposure. These data confirmed the pro-oxidant and genotoxic effects of this fungicide.
Oxidative stress caused by sodium valproate (SV) is known to play a key role in the pathogenesis of brain tissue. The present study was designed to evaluate the protective effect of l-cysteine (LC) on the antioxidants of brain tissue of rats. The animals were divided into six groups: control group 1 was treated with saline as vehicle, groups 2 and 3 were treated with low and high doses of SV (100 and 500 mg/kg, respectively), group 4 was treated with LC (100 mg/kg), and groups 5 and 6 were treated with low-dose SV + LC and high-dose SV + LC, respectively. All the groups were treated orally by gastric tube for 30 successive days. Some antioxidant parameters were determined. Brain tissue (cerebral cortex) of SV-treated animals showed an increase in lipid peroxidation (LPO) and reduction in activity of enzymatic antioxidant and total antioxidant levels. Histopathological examination of cerebral cortex of SV rats showed astrocytic swelling, inflammation, and necrosis. After 4 weeks of the combination treatment of SV and LC daily, results showed significant improvement in the activity of cathepsin marker enzymes and restored the structure of the brain. LC was able to ameliorate oxidative stress deficits observed in SV rats. LC decreased LPO level and was also able to restore the activity of antioxidant enzymes as well as structural deficits observed in the brain of SV animals. The protective effect of LC in SV-treated rats is mediated through attenuation of oxidative stress, suggesting a therapeutic role for LC in individuals treated with SV.
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