Papers by Barbara Plytycz
Advances in Comparative Immunology, 2018
Journal of Herpetology, 1987
Previously undescribed large elongated bodies over the muscles of the groin region (inguinal bodi... more Previously undescribed large elongated bodies over the muscles of the groin region (inguinal bodies) have been found in some representatives of the genus Bufo but not in specimens of the genera Rana, Bombina, and Hyla. Anurans were captured in the field in several localities in southern Poland. On the same day they were killed and examined. Three species of the genus Bufo live in Poland: the common toad, Bufo bufo, the green toad, Bufo viridis, and the relatively rare natterjack, Bufo calamita. In May, the following specimens of the genus Bufo were examined: 7 adults (3 females and 4 males) of B. viridis; 1 adult female and 1 adult male of B. calamita; 8 adults (5 females and 3 males) of B. bufo. In July-August the following specimens of this genus were investigated: 13 adults (7 females and 6 males) and 7 juveniles of B. viridis; 2 adult males of B. calamita; 10 adults (6 females and 4 males) and 3 juveniles of B. bufo. In August, the representatives of both sexes of other anurans living in Poland were also examined, namely the European common frogsRana temporaria (7 adults and 10 juveniles); the frogsRana lessonae (4 adults and 6 juveniles); the yellowbellied toads-Bombina variegata (15 adults and 20 juveniles); the fire-bellied toads-Bombina bombina (5 adults and 7 juveniles); the European tree frogs-Hyla arborea (4 adults and 7 juveniles). In each individual investigated the pelvic part of the abdominal skin and the proximal part of the femoral skin were removed and the groin regions were macroscopically observed. A pair of large elongated bodies (inguinal bodies) supplied by pelvic arteries and veins were visible macroscopically in the groins of each investigated individual of B. viridis (Fig. 1A) and B. calamita. In B. bufo such distinctive structures were never observed (Fig. 1B), however, in a few individuals small accumulations of fat were present in either one or both groins along the pelvic vessels. Similar structures were absent in all investigated representatives of the genera Rana, Bombina, and Hyla. Inguinal bodies of B. viridis and B. calamita, as well as fat accumulations of specimens of B. bufo were easily excised, fixed in Bouin's fluid, routinely embedded in paraffin, serially sectioned, and stained with haematoxylin-eosin or May-Griinwald-Giemsa stain. In B. viridis captured in May, the inguinal bodies were macroscopically brownish and histologically classified as lymphomyeloid bodies (Fig. 2A) since they possesed abundant populations of lymphocytes and granulocytes. In B. viridis captured in July-August, the similar structures of both adults and juveniles were macroscopically yellow and fatty, but serial sections revealed within the fat tissue a few lymphoid foci (Fig. 2B). This suggests that the inguinal bodies of B. viridis undergo conspicuous seasonal changes. In FIG. 1. Inguinal regions of Bufo viridis (Fig. 1A) and Bufo bufo (Fig. 1B). Contours of the inguinal body in the left groin of B. viridis have been outlined (Fig. 1A).
Amphibia-Reptilia, 1993
Yellow-bellied toads were studied in their natural environment in a mountain locality in southeas... more Yellow-bellied toads were studied in their natural environment in a mountain locality in southeastern Poland. 608 specimens were captured, marked by yellow skin autografts placed in different parts of their dorsal surface according to body length, and released. Some of them were recaptured and measured from one to nine years later to estimate their growth and longevity. Yellow-bellied toads grew rapidly in early life; thereafter their growth was very limited. Body size was not an accurate age indicator of an individual of this species. The body length 51-55 mm was maximal in this locality. Yellow-bellied toads were long-lived in nature, some individuals surviving for much more than ten years, and perhaps even more than 20 years. The skeletochronological technique (counting the growth lines in phalangeal cross-sections of the clipped toes of some marked individuals) underestimated the actual age of these animals.
ISJ-Invertebrate Survival Journal, Nov 10, 2011
Earthworm immune-competent cells, celomocytes, are easily retrieved for ex vivo analyses. Celomoc... more Earthworm immune-competent cells, celomocytes, are easily retrieved for ex vivo analyses. Celomocytes consist of amebocytes and species-specific numbers of chloragocyte-derived eleocytes, the latter accumulating free riboflavin in their chloragosome inclusions. Autofluorescent eleocytes are abundant in Eisenia sp., Allolobophora sp., Dendrobaena sp., Dendrodrilus sp., and Octolasion sp., and their numbers and riboflavin contents are affected in species-specific ways by soil quality, as observed by flow cytometry and spectrofluorimetry. The most striking results were obtained in the case of epigeic Dendrodrilus rubidus; in unpolluted soil its riboflavin content was high, but when the earthworm was resident in metalliferous (Pb/Zn-or Ni-polluted) soils, or transferred experimentally from unpolluted to the polluted field soils the riboflavin content was significantly reduced. Such extreme alterations in a cohort of immune-competent cells were not observed in E. andrei, D. veneta, or Al. chlorotica transferred into metalliferous soils. Worms from these three species were also transferred to Zn/Pb/Cd-polluted and unpolluted soils from Southern Poland. It was observed that species-specific changes in riboflavin content occurred not only due to metal pollution, but also other edaphic factors, possibly including organic matter content/quality. Hypothetically, riboflavin status (storage/mobilization) may depend on parasite-immune system balance, which is disrupted by soilderived stressors, including metals.
Central European Journal of Immunology, May 5, 2008
We have shown previously that supplementation of inflammation-inducing factor with a high dose of... more We have shown previously that supplementation of inflammation-inducing factor with a high dose of morphine inhibits intraperitoneal accumulation of exudatory cells in representatives of fish but not in amphibian species, and in some but not all strains of mice. In a case of mice, males were ip injected with zymosan (40 mg/kg, Z group) or with zymosan supplemented with morphine (20 mg/kg, ZM groups). At 4 th hour of peritonitis, the total numbers of intraperitoneal leukocytes (PTLs), and numbers of polymorphonuclear leukocytes (PMNs) were significantly lower in ZM animals than in their Z counterparts in Swiss, C57C3H, Balb/c, and C57BL/6 strains but not in CBA mice. The aim of the present investigations was to monitor the time course of peritonitis (up to 24 hours) in Z and ZM groups of males of four strains. The results fully confirmed the lack of anti-inflammatory action of morphine in CBA strain. In Balb/c, C57BL/6, and Swiss mice, the number of intraperitoneal PMNs was lower in ZM groups during the whole investigated period, being statistically different during the first 12 or 8 hours. In a sharp contrast, the time course of intraperitoneal PMNs accumulation was very similar in Z and ZM groups of CBA mice.
Comparative Biochemistry and Physiology Part C: Toxicology & Pharmacology, 2017
Lumbricid earthworms are often exposed to simultaneous action of various environmental stressors ... more Lumbricid earthworms are often exposed to simultaneous action of various environmental stressors like soil contamination, temperature fluctuation or predators' attacks, which may induce extrusion of coelomocyte-containing coelomic fluid or loss of tail segments. If the injuries are not lethal, renewal of the immune-competent cells and soluble components of coelomic fluid and/or the regeneration of tail segments occurs. The aim of our investigations was to test the hypothesis that exposure of adult earthworms Eisenia andrei to cadmium-polluted soil at room temperature (RT) and/or low temperature (6 o C) have adverse effects on restoration of experimentally depleted coelomocytes or on regeneration of amputated posterior segments. Intact control earthworms and their experimental counterparts subjected to electrostimulationinduced coelomocyte depletion or surgical amputation of posterior segments were maintained either in control soil or in soil spiked with cadmium chloride (500 mg/kg air-dried soil) at RT or 6 o C. Four weeks after the beginning of experiments, cadmium accumulation in earthworm bodies was significantly lower at 6 o C than at room temperature. The numbers of restored cells and fluorophore contents were hardly affected by temperature or cadmium. However, cocoon production was reduced by cadmium and completely abolished at 6°C and regeneration of amputated posterior segments was inhibited in cold but was enhanced by cadmium exposure at RT. Independently on the temperature, the 4-week cadmium exposure of adult earthworms was connected with significantly upregulated expression of Cd-metallothionein (but not of catalase, lysenin and phytochelatin) in coelomocytes.
Applied Soil Ecology, 2016
Earthworms irritated by various noxious stimuli expel coelomic fluid through dorsal pores in the ... more Earthworms irritated by various noxious stimuli expel coelomic fluid through dorsal pores in the body wall. Coelomic fluid contains coelomocytes (amoebocytes and riboflavin-storing eleocytes) and soluble factors responsible for immune functions, thus efficient restoration of the depleted coelomocyte system has an important adaptive value for soil-dwelling species. The aim of present experiments was to compare restoration of coelomocytes and riboflavin (vitamin B2) after experimental expulsion performed in composting earthworms reared for generations in the same laboratory conditions. Dendrobaena veneta,Eisenia andrei and Eisenia fetida were identified according to morphological criteria, species-specific fluorophores, and DNA sequences of mitochondrial COI genes. Analyses (cell counting, flow cytometry, spectrofluorimetry) were conducted on juveniles and adults at the time of electric shockinduced depletion of coelomocyte-containing coelomic fluid and then three weeks later in worms recovering their immune systems, and in their untreated counterparts. Electric shock induced expulsion of approximately 90% amoebocytes, eleocytes, and riboflavin, followed by their gradual restoration that was still in progress 3 weeks later. Amoebocytes were restored faster than riboflavin and eleocytes. Riboflavin content was higher in restored than senescent eleocytes of adult worms. Process of restoration was faster in juveniles than in adults and in the former was connected with accelerated gain in body mass. Senescent coelomocytes of adult earthworms easily undergo fragmentation, while it was seldom observed in coelomocytes of juveniles and in newly formed coelomocytes replacing expelled cells in both adult and juvenile worms. Molecular basis of these phenomena are worth further elucidation.
The Journal of Experimental Zoology, 1998
The effects of ambient temperature (22 degrees C/10 degrees C) and season (summer/winter) on anur... more The effects of ambient temperature (22 degrees C/10 degrees C) and season (summer/winter) on anuran skin allograft and xenograft rejection was tested in frogs (Rana temporaria and R. esculenta) and toads (Bufo bufo, Bombina variegata, and Bombina bombina). Mean graft survival times were significantly prolonged at the low temperature in a species-specific manner, the edible frog (R. esculenta) being the most sensitive and the common toad (Bufo bufo) relatively resistant. Allografts were more temperature-dependent than xenografts; in the latter case, temperature sensitivity was specific to each donor-host combination. Rejection of second-set grafts in R. esculenta was accelerated both in warmth and in cold, but second-set grafts were less temperature-sensitive than sensitizing ones. Both in summer and in winter, R. esculenta rejected allografts promptly at 22 degrees C but slowly at 10 degrees C. In both seasons, Bombina variegata kept at 22 degrees C rejected allografts in a chronic manner. This indicates that amphibian transplantation immunity depends on the donor-host genetic disparity and ambient temperature but is independent of season.
Inflammation
The initial phase of zymosan-induced peritonitis involves an increase of vascular permeability (p... more The initial phase of zymosan-induced peritonitis involves an increase of vascular permeability (peak at 30 min) that is correlated with high levels of vasoactive eicosanoids, namely, prostaglandins (PGI2 and PGE2) of cyclooxygenase-1 origin (as estimated by RT-PCR) and cysteinyl-leukotrienes. Previously, we showed that the increase of vascular permeability can be attributed only partially to mast cells and their histamine, as seen in mast cell-deficient WBB6F1-W/Wv mice. Thus we aimed to identify the major cellular source(s) that mediate vasopermeability, as well as particular vasoactive mediators operating in this model. For this purpose, some mice were selectively depleted of either peritoneal macrophages or mast cells, and/or they were treated with several pharmacologic inhibitors of cyclooxygenase- and lipoxygenase-metabolic pathways. More-over, macrophage-depleted mast cell-deficient WBB6F1-W/Wv mice and their controls (+/+) were used. The macrophage depletion always caused a p...
Developmental and comparative immunology, Jan 8, 2015
Earthworm decerebration causes temporary inhibition of reproduction which is mediated by certain ... more Earthworm decerebration causes temporary inhibition of reproduction which is mediated by certain brain-derived neurohormones; thus, cocoon production is an apposite supravital marker of neurosecretory center functional recovery during brain regeneration. The core aim of the present study was to investigate aspects of the interactions of nervous and immune systems during brain regeneration in adult Dendrobaena veneta (Annelida; Oligochaeta). Surgical brain extirpation was combined, either with (i) maintenance of immune-competent coelomic cells (coelomocytes) achieved by surgery on prilocaine-anaesthetised worms or (ii) prior extrusion of fluid-suspended coelomocytes by electrostimulation. Both brain renewal and cocoon output recovery were significantly faster in earthworms with relatively undisturbed coelomocyte counts compared with individuals where coelomocyte counts had been experimentally depleted. These observations provide empirical evidence that coelomocytes and/or coelomocyte...
Life Sciences, 2011
We investigated the effects of riboflavin (vitamin B2) on the kinetics of zymosan-induced periton... more We investigated the effects of riboflavin (vitamin B2) on the kinetics of zymosan-induced peritonitis in three strains of mice. Main methods: Peritonitis was induced in males of C57BL/6J, BALB/c and CBA mice by intraperitoneal injection of zymosan (40 mg/kg) or zymosan supplemented with riboflavin (50 mg/kg). During the first 45 min of inflammation the pain symptoms were scored. At the selected time points (4, 6, 8, 10, 24, and 30 h) the mice were sacrificed and peritoneal exudates were retrieved. Leukocytes, among them polymorphonuclear cells (PMNs) and macrophages (Mac3 + cells) were counted. Levels of inducible nitric oxide synthase (iNOS) were measured in cell pellets while supernatants were used for measurements of nitric oxide, cytokine/chemokines (IL-6, IL-10, MCP-1, IFNγ, TNF-α, and IL-12p70), and matrix metalloproteinase-9 (MMP-9). Key finding: A riboflavin ip injection induced pain symptoms itself, but reduced zymosan-induced pain in C57BL/6J and CBA strains of mice when coinjected with zymosan. In comparison with the mice injected with zymosan only, riboflavin coinjection prolonged inflammation in C57BL/6J mice due to prolonged macrophage accumulation; inhibited peritoneal leukocytes (PTL) accumulation in BALB/c due to inhibited influx of macrophages and PMNs; and inhibited PTL accumulation in CBA mice due to delayed PMN influx. These effects corresponded with the delayed (C57BL/6J) or inhibited (BALB/c and CBA) expression of iNOS in PTL lysates, and with the prolonged (C57BL/6) or inhibited (BALB/c) intraperitoneal accumulation of MMP-9. Moreover, cytokine accumulation was affected in a strain-specific way. Significance: Riboflavin is antinociceptive during yeast-induced peritonitis, but its anti-inflammatory effects are strain-specific.
Life Sciences, 2012
We compared the effects of riboflavin pre-injection, co-injection and post-injection on several s... more We compared the effects of riboflavin pre-injection, co-injection and post-injection on several symptoms of zymosan-induced peritonitis in male Swiss mice. Additionally, the effects of i.p. injection of riboflavin itself were elucidated. Peritonitis was induced in Swiss mice (50 animals) by i.p. zymosan (Z; 40mg/kg) injection. Riboflavin (R; 0, 20, 50, or 100mg/kg) was applied either alone or in combination with zymosan. In the latter case riboflavin was administered either together with zymosan (R group), or 30min before zymosan (R-Z group), or 1h later (Z-R group). The nociceptive response was evaluated by counting body writhes. The peritoneal exudates retrieved 4h after the R or Z injection were analyzed for the numbers and apoptosis of polymorphonuclear leukocytes (PMNs), and levels of metalloproteinase 9 (MMP-9), nitric oxide, and inflammatory cytokines, IL-12p70, TNFα, MCP-1, IL-6, IL-10, IFNγ. Riboflavin itself induced nociceptive-related body writhes and a moderate inflammatory response manifested by PMN influx and the release of some cytokines and MMP-9. In contrast, antinociceptive properties of riboflavin were significant in the ZR group co-injected with the lowest dose of riboflavin (ZR20). At the 4th hour of zymosan-induced peritonitis an intraperitoneal accumulation of PMNs was decreased in the riboflavin-treated groups and cytokine profiles were modified according to riboflavin dose and the time of injection. Riboflavin itself induces low-grade nociception and inflammation while its effects on zymosan-induced inflammation are dependent on the dose and time of its application: either before or during inflammation.
Trends in Immunology, 2002
Ex vivo staining of metastatic lymph nodes by class I major histocompatibility complex tetramers ... more Ex vivo staining of metastatic lymph nodes by class I major histocompatibility complex tetramers reveals high numbers of antigen-experienced tumorspecific cytolytic T lymphocytes. J. Exp. Med. 188, 1641-1650 3 Zippelius, A. et al. (2002) Thymic selection generates a large T-cell pool recognizing a self-peptide in humans.
Photochemistry and Photobiology, 2006
; the latter are filled with numerous granules. We have previously shown that eleocytes of severa... more ; the latter are filled with numerous granules. We have previously shown that eleocytes of several (but not all) earthworm species exhibit strong autofluorescence detectable by fluorescent microscopy and flow cytometry. In the present article, the molecular origin of eleocytes autofluorescence was elucidated in coelomocytes expelled via dorsal pores in the integument of Eisenia fetidu subjected to electric shock (1 min at 4.5 V). Spectrofluorometry (excitation and emission spectra and fluorescence lifetime), together with HPLC analysis of coelomocyte suspensions and supernatants, indicated that riboflavin but not FMN (flavin mononucleotide) or FAD (flavin-adenine dinucleotide) is the main fluorophore responsible for eleocyte fluorescence in this species. Additionally, lipofuscins are suspected to participate in this phenomenon.
Pedobiologia, 2003
Coelomocytes (extruded via dorsal pores of earthworms subjected to 5V electric shock) of adult Al... more Coelomocytes (extruded via dorsal pores of earthworms subjected to 5V electric shock) of adult Allolobophora chlorotica, Lumbricus terrestris, Dendrobaena veneta and Eisenia fetida consisted of species-specific proportions of hyaline amoebocytes, granular amoebocytes, and eleocytes. In all species a significant increase of granular amoebocytes with a parallel decrease of hyaline amoebocytes was recorded by the end of Spring and beginning of Summer, both in animals from the natural environment and in those kept in the laboratory at the room temperature. It indicates that coelomocyte proliferation/maturation is dependent mainly on the annual endogenous rhythm, which is relatively stable even at atypical ambient temperatures. During four successive seasons viability and functions (plastic adherence or neutral red pinocytosis) were recorded in coelomocytes of A. chlorotica assayed at various in vitro temperatures. In coelomocytes retrieved from animals freshly collected from their natural environment, coelomocyte activity was better at low temperatures (0 °C and 10 °C) during the Winter, while during other seasons (especially in the Summer) it was best at 22 °C. Such seasonal effects were abolished in coelomocytes from animals tested after one-month acclimation to the room temperature, as in all seasons cell functions were best at 22 °C. These observations indicate that earthworm coelomocytes can thermally adapt. This phenomenon was also recorded in L. terrestris, D. veneta and E. fetida. In conclusion, earthworm coelomocyte proliferation/maturation seems to be endogenously controlled while coelomocyte activity seems to be more plastic and subjected to thermal adaptation.
Pedobiologia, 2011
... Biomarkers 8, 229-239.34 Page 14 of 23 Ac ce pte d M an usc rip t 14 Hankard, PK,Svendsen, C.... more ... Biomarkers 8, 229-239.34 Page 14 of 23 Ac ce pte d M an usc rip t 14 Hankard, PK,Svendsen, C., Wright, J., Wienberg, C., Fishwick, SK, Spurgeon, DJ, 1 Weeks, JM, 2004. ... In vitro fusion of newt peritoneal 19 macrophages. J. Exp. Zool. ...
Pedobiologia, 2011
... Agnieszka Irena Mazur a , Malgorzata Klimek a , A. John Morgan b , Barbara Plytycz a , Corres... more ... Agnieszka Irena Mazur a , Malgorzata Klimek a , A. John Morgan b , Barbara Plytycz a , Corresponding Author Contact Information , E-mail The ... Mayeno et al., 1992 AN Mayeno, KJHamann and GJ Gleich, Granule-associated flavin adenine dinucleotide (FAD) is responsible ...
Journal of Experimental Zoology Part A: Ecological Genetics and Physiology, 2013
Supravital species identification of morphologically similar syntopic earthworms inhabiting dung ... more Supravital species identification of morphologically similar syntopic earthworms inhabiting dung and compost heaps or those from commercial cultures is difficult. The aim of the studies was to find out non-invasive species-specific markers for proper segregation of earthworm species from a dense mixed colony of waste decomposers. Worms were segregated according to external characteristics into Eisenia andrei, Eisenia fetida, and Dendrobaena veneta, and left for reproduction and analysis of non-invasively retrieved coelomocyte-containing coelomic fluid and/ or species-specific partial sequences of cytochrome c oxidase subunit I (COI) gene in DNA extracted from amputated tail tips of adults and their offspring. Flow cytometric analysis of coelomocyte samples revealed that amount of nuclear DNA increases in order D. veneta ( E. andrei < E. fetida, and intensity of eleocyte-derived fluorescence is lower in D. veneta than in Eisenia spp. Spectrofluorimetry of coelomocyte lysates revealed that the amount of eleocyte-stored riboflavin is significantly lower in coelomocyte lysates from D. veneta than from Eisenia spp., and the emission peak of X-fluorophore is much more distinct in D. veneta than in Eisenia spp. Coelomic fluid of E. andrei exhibits a very distinct spectra of MUG fluorophore which are absent in D. veneta and in the majority of E. fetida, while some E. fetida possess MUG-like fluorophore. Sequences of the COI gene in the DNA of the worms from the mixed colony and their offspring confirmed species identity. In conclusion, species-specific coelomocyte-derived markers may be a useful complement to morphological and DNA-based taxonomy during studies on syntopic earthworms.
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Papers by Barbara Plytycz