• Mode of action of FMRFamide-like peptide on drosophila body wall muscle conractions

      Milakovic, Maja; Department of Biological Sciences (Brock University, 2011-03-08)
      Neuropeptides are the largest group of signalling chemicals that can convey the information from the brain to the cells of all tissues. DPKQDFMRFamide, a member of one of the largest families of neuropeptides, FMRFamide-like peptides, has modulatory effects on nerve-evoked contractions of Drosophila body wall muscles (Hewes et aI.,1998) which are at least in part mediated by the ability of the peptide to enhance neurotransmitter release from the presynaptic terminal (Hewes et aI., 1998, Dunn & Mercier., 2005). However, DPKQDFMRFamide is also able to act directly on Drosophila body wall muscles by inducing contractions which require the influx of extracellular Ca 2+ (Clark et aI., 2008). The present study was aimed at identifying which proteins, including the membrane-bound receptor and second messenger molecules, are involved in mechanisms mediating this myotropic effect of the peptide. DPKQDFMRFamide induced contractions were reduced by 70% and 90%, respectively, in larvae in which FMRFamide G-protein coupled receptor gene (CG2114) was silenced either ubiquitously or specifically in muscle tissue, when compared to the response of the control larvae in which the expression of the same gene was not manipulated. Using an enzyme immunoassay (EIA) method, it was determined that at concentrations of 1 ~M- 0.01 ~M, the peptide failed to increase cAMP and cGMP levels in Drosophila body wall muscles. In addition, the physiological effect of DPKQDFMRFamide at a threshold dose was not potentiated by 3-lsobutyl-1-methylxanthine, a phosphodiesterase inhibitor, nor was the response to 1 ~M peptide blocked or reduced by inhibitors of cAMP-dependent or cGMP-dependent protein kinases. The response to DPKQDFMRFamide was not affected in the mutants of the phosholipase C-~ (PLC~) gene (norpA larvae) or IP3 receptor mutants, which suggested that the PLC-IP3 pathway is not involved in mediat ing the peptide's effects. Alatransgenic flies lacking activity of calcium/calmodul in-dependent protein kinase (CamKII showed an increase in muscle tonus following the application of 1 JlM DPKQDFMRFamide similar to the control larvae. Heat shock treatment potentiated the response to DPKQDFMRFamide in both ala1 and control flies by approximately 150 and 100 % from a non heat-shocked larvae, respectively. Furthermore, a CaMKII inhibitor, KN-93, did not affect the ability of peptide to increase muscle tonus. Thus, al though DPKQDFMRFamide acts through a G-protein coupled FMRFamide receptor, it does not appear to act via cAMP, cGMP, IP3, PLC or CaMKl1. The mechanism through which the FMRFamide receptor acts remains to be determined.
    • Modulation of muscle contraction by a FMRFamide-related peptide

      Quigley, Patricia A.; Department of Biological Sciences (Brock University, 1995-11-04)
      A FMRFamide-like neuropeptide with the sequence "DRNFLRF-NH2" was recently isolated from pericardial organs of crayfish (Mercier et aI., Peptides, 14, 137-143, 1993). This neuropeptide, referred to as "DF2'" has already been shown to elicit cardioexcitation and to enhance synaptic transmission at neuromuscular junctions. Possible effects ofDF2 on muscle were investigated using superficial extensor muscles of the abdomen of the crayfish, Procambarus clar/ai. These muscles are of the tonic type and generate slow contractions that affect posture. DF2, at concentrations of 10-8 M or higher, increased muscle tonus and induced spontaneous, rhythmic contractions. These effects were antagonized by 5 rnM Mn2+ but not by lO-7M tetrodotoxin (TTX). Thus, they represent direct actions on muscle cells (rather than effects on motor neurons) and are likely to involve calcium influx. In contrast, deep abdominal extensor muscles, responsible for rapid swimming movements, and superficial flexor muscles do not generate contractions in response to the peptide. 2 Spontaneous contractions were also induced in the superficial extensor muscles by decreasing the temperature to II-13°C. Such contractions were also TTX-insensitive and they were antagonized by adding calcium channel blockers (Mn2+, Cd2+ or Ni2+) or by removing calcium from the bathing solution. This suggests that the spontaneous contractions depend on an influx of calcium from the extracellular solution. N-type and L-type voltage dependent calcium channel blockers did not reduce the effect of the peptide or the spontaneous contractions suggesting that calcium influx is not through N- or L-type calcium channels.
    • Molecular cloning and restriction endonuclease analysis of bovine adenovirus type 3

      Elgadi, Mabrouk Mohammed.; Department of Biological Sciences (Brock University, 1993-07-09)
      Bovine adenovirus type 3 (BAV3) is a medium size DNA virus that causes respiratory and gastrointestinal disorders in cattle. The viral genome consists of a 35,000 base pair, linear, double-stranded DNA molecule with inverted terminal repeats and a 55 kilodalton protein covalently linked to each of the 5' ends. In this study, the viral genome was cloned in the form of subgenomic restriction fragments. Five EcoRI internal fragments spanning 3.4 to 89.0 % and two Xb a I internal fragments spanning 35.7 to 82.9 % of the viral genome were cloned into the EcoRI and Xbal sites of the bacterial vector pUC19. To generate overlap between cloned fragments, ten Hi n dIll internal fragments spanning 3.9 to 84.9 and 85.5 to 96% and two BAV3 BamHI internal fragments spanning 59.8 to 84.9% of the viral genome were cloned into the HindllI and BamHI sites of pUC19. The HindlII cloning strategy also resulted in six recombinant plasmids carrying two or more Hi ndII I fragments. These fragments provided valuable information on the linear orientation of the cloned fragments within the viral genome. Cloning of the terminal fragments required the removal of the residual peptides that remain attached to the 5' ends of the genome. This was accomplished by alkaline hydrolysis of the DNA-peptide bond. BamH I restriction fragments of the peptide-free DNA were cloned into pUC19 and resulted in two plasmids carrying the BAV3 Bam HI terminal fragments spanning 0 to 53.9% and 84.9 to 100% of the viral genome.
    • Molecular cloning restriction enzyme analysis, bovine adenovirus type 2 genome

      Salmon, Kirsty Anne.; Department of Biological Sciences (Brock University, 1993-07-09)
      Adenoviruses are non-enveloped icosahedral-shaped particles which possess a double-stranded DNA genome. Currently, nearly 100 serotypes of adenoviruses have been identified, 48 of which are of human origin. Bovine adenoviruses (BAVs), causing both mild respiratory and/or enteral diseases in cattle, have been reported in many countries all over the world. Currently, nine serotypes of SAVs have been isolated which have been placed into two subgroups based on a number of characteristics which include complement fixation tests as well as the ability to replicate in various cell lines. Bovine adenovirus type 2 (BAV2), belonging to subgroup I, is able to cause pneumonia as well as pneumonic-like symptoms in calves. In this study, the genome of BAV2 (strain No. 19) was subcloned into the plasmid vector pUC19. In total, 16 plasmids were constructed; three carry internal San fragments (spanning 3.1 to 65.2% ), and 10 carry internal Pstl fragments (spanning 4.9 to 97.4%), of the viral genome. Each of these plasmids was analyzed using twelve restriction endonucleases; BamHI, CiaI, EcoRl, HiOOlll, Kpnl, Noll, NS(N, Ps~, Pvul, Saj, Xbal, and Xhol. Terminal end fragments were also cloned and analyzed, sUbsequent to the removal of the 5' terminal protein, in the form of 2 BamHI B fragments, cloned in opposite orientations (spanning 0 to 18.1°k), and one Pstll fragment (spanning 97.4 to 1000/0). These cloned fragments, along with two other plasmids previously constructed carrying internal EcoRI fragments (spanning 20.6 to 90.5%), were then used to construct a detailed physical restriction map using the twelve restriction endonucleases, as well as to estimate the size of the genome for BAV2(32.5 Kbp). The DNA sequences of the early region 1 (E1) and hexon-associated gene (protein IX) have also been determined. The amino acid sequences of four open reading frames (ORFs) have been compared to those of the E1 proteins and protein IX from other Ads.
    • The nesting biology of Ceratina (Hymenoptera: Apidae) in the Niagara Region : new species, nest site selection and parasitism

      Vickruck, Jess; Department of Biological Sciences (Brock University, 2010-10-27)
      One of the most common bee genera in the Niagara Region, the genus Ceratina (Hymenoptera: Apidae) is composed of four species, C. dupla, C. calcarata, the very rare C. strenua, and a previously unknown species provisionally named C. near dupla. The primary goal of this thesis was to investigate how these closely related species coexist with one another in the Niagara ~ee community. The first necessary step was to describe and compare the nesting biologies and life histories of the three most common species, C. dupla, C. calcarata and the new C. near dupla, which was conducted in 2008 via nest collections and pan trapping. Ceratina dupla and C. calcarata were common, each comprising 49% of the population, while C. near dupla was rare, comprising only 2% of the population. Ceratina dupla and C. near dupla both nested more commonly in teasel (Dipsacus sp.) in the sun, occasionally in raspberry (Rubus sp.) in the shade, and never in shady sumac (Rhus sp.), while C. calcarata nested most commonly in raspberry and sumac (shaded) and occasionally in teasel (sunny). Ceratina near dupla differed from both C. dupla and C. calcarata in that it appeared to be partially bivoltine, with some females founding nests very early and then again very late in the season. To examine the interactions and possible competition for nests that may be taking place between C. dupla and C. calcarata, a nest choice experiment was conducted in 2009. This experiment allowed both species to choose among twigs from all three substrates in the sun and in the shade. I then compared the results from 2008 (where bees chose from what was available), to where they nested when given all options (2009 experiment). Both C. dupla and C. calcarata had the same preferences for microhabitat and nest substrate in 2009, that being raspberry and sumac twigs in the sun. As that microhabitat and nest substrate combination is extremely rare in nature, both species must make a choice. In nature Ceratina dupla nests more often in the preferred microhabitat (sun), while C. calcarata nests in the preferred substrate (raspberry). Nesting in the shade also leads to smaller clutch sizes, higher parasitism and lower numbers of live brood in C. calcarata, suggesting that C. dupla may be outcompeting C. calcarata for the sunny nesting sites. The development and host preferences of Ceratina parasitoids were also examined. Ceratina species in Niagara were parasitized by no less than eight species of arthropod. Six of these were wasps from the superfamily Chalcidoidea (Hymenoptera), one was a wasp from the family Ichneumonidae (Hymenoptera) and one was a physogastric mite from the family Pyemotidae (Acari). Parasites shared a wide range of developmental strategies, from ichneumonid larvae that needed to consume multiple Ceratina immatures to complete development, to the species from the Eulophidae (Baryscapus) and Encyrtidae (Coelopencyrtus), in which multiple individuals completed development inside a single Ceratina host. Biological data on parasitoids is scarce in the scientific literature, and this Chapter documents these interactions for future research.
    • Nestmate recognition in the large carpenter bee, Xylocopa virginica /

      Peso, Marianne.; Department of Biological Sciences (Brock University, 2008-06-15)
      Abstract Many species of social insects have the ability to recognize their nestmates. In bees, sociality is maintained by bees that recognize which individuals should be helped and which should be hanned in order to maximize fitness (either inclusive or individual) (Hamilton 1964; Lin and Michener 1972). Since female bees generally lay eggs in a single nest, it is highly likely that bees found cohabitating in the same nest are siblings. According to the kin selection hypothesis, individuals should cooperate and avoid aggression with same sex nestmates (Hamilton 1964). However, in opposite sex pairs that are likely kin, aggression should increase among nestmates as an expression of inbreeding avoidance (Lihoreau et al. 2007). Female bees often guard nest entrances, recognizing and excluding foreign conspecific females that threaten to steal nest resources (Breed and Page 1991). Conversely, males that aggressively guard territories should avoid aggression towards other males that are likely kin (Shellman-Reeve and Gamboa 1984). In order to test whether Xy/ocopa virginica can distinguish nestmates from non-nestmates, circle tube testing arenas were used. Measures of aggression, cooperation and tolerance were evaluated to detennine the presence of nestmate recognition in this species. The results of this study indicate that male and female X virginica have the ability to distinguish nestmates from non-nestmates. Individuals in same sex pairs demonstrated increased pushing, biting, and C-posturing when faced with non-nestmates. Males in same sex pairs also attempted to pass (unsuccessfully) nOIl-nestmates more often than ncstmates, suggesting that this behaviour may be an cxpression of dominancc in males. Increased cooperation exemplified by successful passes was not observed among nestmates. However, incrcased tolerance in the [onn of head-to-head touching was observed for nestmates in female same sex and opposite sex pairs. These results supported the kin selection hypothesis. Moreover, increased tolerance among opposite sex non-nestmates suggested that X virginica do not demonstrate inbreeding avoidance among nestmates. 3 The second part of this study was conducted to establish the presence and extent of drifting, or travelling to different nests, in a Xylocopa virgillica population. Drifting in flying Hymenoptera is reported to be the result of navigation error and guard bees erroneously admitting novel individuals into the nest (Michener 1966). Since bees in this study were individually marked and captured at nest entrances, the locations where individuals were caught allowed me to determine where and how often bees travelled from nest to nest. Ifbees were captured near their home nests, changing nests may have been deliberate or explained by navigational error. However, ifbees were found in nests further away from their homes, this provides stronger evidence that flying towards a novel nest may have been deliberate. Female bees are often faithful to their own nests (Kasuya 1981) and no drifting was expected in female X virginica because they raise brood and contribute to nest maintenance activities. Contrary to females, males were not expected to remain faithful to a single nest. Results showed that many more females drifted than expected and that they were most often recaptured in a single nest, either their home nest or a novel nest. There were some females that were never caught in the same nest twice. In addition, females drifted to further nests when population density was low (in 2007), suggesting they seek out and claim nesting spaces when they are available. Males, as expected, showed the opposite pattern and most males drifted from nest to nest, never recaptured in the same location. This pattern indicates that males may be nesting wherever space is available, or nesting in benches nearest to their territories. This study reveals that both female and male X virginica are capable of nestmate recognition and use this ability in a dynamic environment, where nest membership is not as stable as once thought.
    • Orientation Preference and Behavioural Thermoregulatory Coordination in Pogona vitticeps.

      Black, Ian; Department of Biological Sciences
      Regulating body temperature is a critical function for vertebrates and many invertebrates. Vertebrates that rely on ambient temperature as a heat source (ectotherms) make use of multiple voluntary and involuntary behaviours to thermoregulate, including body orientation. The purpose of this work was to examine orientation behaviour in bearded dragons (Pogona vitticeps) as well as possible coordination between thermoregulatory responses. Both adult and two week old neonatal bearded dragons were placed in a thermal gradient and left to thermoregulate behaviourally in order to observe the presence of a thermoregulatory orientation preference. Since a thermal orientation bias could manifest from a preference for favoring rostral versus caudal traits, animals were subsequently subjected to a separate experiment, in which either heat exchange across the tail or respiratory heat exchange from gaping was inhibited. Changes in thermoregulatory behaviour were then observed in the manipulated bearded dragons in order to assess the potential contributors to thermal preferences as well as isolate potential coordination between behaviours. Both adult and two week old neonatal dragons displayed a non-random orientation preference for facing a heat source that strengthened with time exposed to a thermal gradient, supporting this behaviour as a thermoregulatory response. Bearded dragons also exhibited changes in orientation preference and gaping behaviour when the tail was insulated and when gaping was inhibited, but no changes to selected ambient temperature, indicating that the changes to gaping and orientation behaviours are compensatory responses that support the presence of coordination between thermoregulatory behaviours.
    • Oxygen exchange in galls of Eurosta solidaginis (Diptera: Tephritidae)

      Long, Qian; Department of Biological Sciences (Brock University, 2013-01-24)
      Hypoxia in plant tissue should affect animals living within. Gallmakers stimulate their plant hosts to produce the gall they inhabit and feed on, and also influence the gall phenotype for other adaptations, such as defense against predators. The potential for hypoxia in galls of Eurosta solidaginis was studied in the context of potential adaptations to gall oxygen level, using a combination of direct measurement, mathematical modelling, and respirometry on both gallmakers and hosts. Modelling results suggested mild hypoxia tolerable to the larva persists for most of the growth season, whereas more severe hypoxia may occur earlier in fully-grown young galls. Field data from one of the two years studied showed hypoxia more severe than expected, and coincided with adverse weather conditions and high larval mortality. The hypoxia may be related to host response to adverse weather. Whether hypoxia directly caused larval mortality requires further study.
    • Palaeopigment accumulation and implcations for paleolimnology over the lost century for two Ontario lakes

      Han, Xiaozhong.; Department of Biological Sciences (Brock University, 1993-07-09)
      Crawford Lake is a meromictic lake, which is 24 m deep and has an area of 2.5 ha, and has never been reported to have mixed below 16 m. Lady Evelyn Lake, which became a reservoir when a dam was built in 1916, is dimictic with a maximum depth of about 35 m. 1 My research proved that both native chlorophylls and the ratio of chlorophyll derivatives to total carotenoids were better preserved in the shallower lake (Crawford Lake) because it was meromictic. Thus the anaerobic conditions in Crawford Lake below 16 m (monimolimnion) provide excellent conditions for pigment preservation. Under such conditions, the preservation of both chlorophylls and carotenoids, including oscillaxanthin and myxoxanthophyll, are extremely good compared with those of Lady Evelyn Reservoir, in which anaerobic conditions are rarely encountered at the mud-water interface. During the period from 1500 to 1900 A. D. in Crawford Lake, the accumulation rates of oscillaxanthin and myxoxanthophyll were extremely high, but those of chlorophyll derivatives and total carotenoids were relatively low. This was correlated with the presence of a dense benthic mat of cyanobacteria near the lake's chemocline. Competition for light between the deep dwelling cyanobacteria and overlying phytoplankton in this meromictic lake would have been intensified as the lake became more and more eutrophic (1955-1991 A. D.). During the period from 1955 to 1991 A. D., the accumulation rates of chlorophyll derivatives and total carotenoids in the sediment core from Crawford Lake (0-7.5 cm, 1955-present) increased. During this same period, the accumulation rates of cyanobacterial pigments (Le. oscillaxanthin and myxoxanthophyll) declined as the lake became more eutrophic. Because the major cyanobacteria in Crawford Lake are benthic mat forming Lyngbya and Oscillatoria and not phytoplankton, eutrophication resulted in a decline of the mat forming algal pigments. This is important because in previous palaeolimnological studies the concentrations of oscillaxanthin and myxoxanthophyll have been used as correlates with lake trophic levels. The results of organic carbon a13c analysis on the Crawford Lake sediment core supported the conclusions from the pigment study as noted above. High values of a13c at the depth of 34-48 cm (1500-1760 A. D.) were related to a dense population of benthic Oscillatoria and Lyngbya living on the bottom of the lake during that period. The Oscillatoria and Lyngbya utilized the bicarbonate, which had a high a 13C value. Very low values were found at 0-7 cm in the Crawford sediment core. At this time phytoplankton was the main primary producer, which enriched 12C by photosynthetic assimilation.
    • Paleoindicators of meromixis

      Cheek, Michael Ross.; Department of Biological Sciences (Brock University, 1979-07-09)
      This study was undertaken to ascertain whether meromictic lakes could be differentiated from holomictic lakes on the basis of their surficial profundal sediments. Surface sediment cores (15 cm long) were collected from both the littoral and profundal zones of four meromictic and six holomictic lakes and analyzed for total number of fossil chironomid headcapsu~es, chlorophyll and carotenoid degradation products as well as \ iron and manganese concentrations. Littoral and profundal comparisons of the surface sediments were made between the two lake types using the Mann-Whitney U test. Iron, manganese and the iron to manganese ratio in the littoral sediments of meromictic lakes were significantly lower than those found in the littoral sediments of holomictic lakes. The observed differences are believed to represent an artifact of the significantly higher carbonate concentrations found in three of the four meromictic lakes studied. Profundal and littoral to profundal ratio comparison between holomictic and meromictic lakes suggest that the significantly lower iron and higher carotenoid concentrations in meromictic profundal sediments were a con~equence of meromixis. However, the overlap in distribution exhibited by both iron and carotenoid degradation products between the two lake types was sufficiently large in this study to nullify their use as a means of differentiating meromictic from holomictic lakes. A long core (4.25 m) was removed from the deepest part of the meromictic Crawford Lake (Ontario), sectioned at 5 cm intervals, and analyzed to assess when meromixis occurred, based on its fossil record. Temporal changes in the total number of chironomid headcapsules, and chlorophyll and carotenoid sediment degradation products were closely correlated with organic matter, indicating in my opinion that extensive redeposition of littoral chironomid headcapsules in the profundal zone has occurred. Temporal variations in carotenoid degradation products, in response to changes in organic matter, obscured increased preservation that may have occurred as a consequence of meromixis. Temporal variations in iron and manganese suggest that relatively stable redox conditions have existed throughout most of the lake's history. Therefore it would appear that Crawford Lake has been meromictic since its inception.
    • Paleolimnology of selected Precambrian Shield lakes /

      Smol, J. P. (John P.); Department of Biological Sciences (Brock University, 1979-06-29)
      The analysis of fossil diatoms and Mallomonas assemblages in a 2.85 m sediment core revealed that a series of distinct floristic changes have occurred in the development of Found Lake, a small Shield lake in southern Ontario. Climatic and vegetational changes in the lake's watershed were closely associated with successional changes in the lake's biota. Nutrients released by the deciduous component of the Found Lake watershed appeared to be especially important in determining diatom and Mallomonas standing crop. The top 20 cm of sediment of 3 ,Shield lakes was then investigated using close interval (1 cm) analyses of diatoms, Mallomonas scales, pollen grains and sedimentary phosphorus. Found and Jake Lake are adjacent to Highway 60, whereas Delano Lake has been undisturbed and was used as a control. Dramatic changes in the diatom and Mallomonas communities were recorded in the Found and Jake Lake stratigraphies and could be closely associated with known historical events. Increased turbidity and nutrient enrichment were believed responsible for these successional changes. In addition, diatom and Mallomonas standing crop increased substantially following road construction in Found Lake's drainage basin. Meanwhile, no. sharp changes in diatom or Mallomonas communities were recorded in the recent sediments of the control (Delano) lake. The use£ulness o£ Synuracean scales as paleoindicators, as well as the importance o£ sectioning cores at close intervals during transition periods in a laker's development, was stressed.
    • The pancreatic islet system of the rock bass /|nby Edward Francis Squires. -- 260 St. Catharines, Ont. : [s. n.],

      Squires, Edward Francis.; Department of Biological Sciences (Brock University, 1972-07-09)
      The endocrine pancreas of the rock bass (Ambloplites rupestris) was examined by light and electron microscopy. Two cell types with staining properties similar to mammalian A and B cells, and a third, non-staining cell type were found in the spherical pancreatic islets that were surrounded by a connective tissue capsule and embedded in two small masses of exocrine tissue. From an analysis of the ultrastructure of the A and B cells, a secretory cycle for each of these cell types was proposed. The secretory cycle of the A cell consisted of three well defined stages: (1) A cell production stage: during which A granule formation occurred in the sacs of the Golgi apparatus and the cell was characterized by the presence of numerous secretory granules, some elements of lamellar endoplasmic reticulum, and a homogeneously granular nucleus. The cytoplasm contained few distended cisternae, variable numbers of free ribosomes, microtubules and small vesicles. (2) A cell release stage: during which the release of A granules occurred and the cell usually contained several large distended cisternae and variable numbers of secretory granules. Granule release mechanisms included exocytosis, by which individual granules were released into the extracellular space after their membranes fused with the plasmalemma, and emiocytosis, by which one or more granules were released into a large cisterna whose membrane fused with the plasmalemma and formed a pore through which the cisternal contents passed out of the cell. (3) A cell reorganization stage: during which the changeover from the release stage to the production stage occurred and the reorganization of organelles and membrane structures took place. The cell contained few secretory granules and numerous small endoplasmic reticular cisternae. The cytoplasm exhibited less electron density than either of the other two stages. The A granule after formation underwent a series of morphological changes which were described in four numerically identified phases. The secretory cycle of the B cell consisred of two stages: (1) B cell production stage: during which the B granule formation occurred in the sacs of the Go1gi apparatus. The cell was characterized by an irregular outline, the presence of numerous secretory granules, and an irregularly shaped nucleus which contained variable amounts of clumped chromatin. The cytoplasm contained moderate amounts of lamellar endoplasmic reticulum studded with ribosomes, several small vesicles, and an active Go1gi apparatus. (2) B cell release stage: during which the release of B granules occurred. The cell contained a rounded nucleus with dispersed chromatin, several distended endoplasmic reticular cisternae and a variable number of secretory granules. Granule release occu~ by emiocytosis and exocytosis similar to that found for the A cell.
    • Parental care in the common tern (Sterna Nirundo): sexual roles in a monogamous seabird

      Wiggins, David A.; Department of Biological Sciences (Brock University, 1984-07-09)
      The parental behaviour of male and female Common Terns (Sterna hirundo) was documented throughout two breeding seasons at a colony near Port Colborne , Ontario. Thirteen and fourteen pairs of terns were chosen for intensive study in 1982 and 1983, respectively. The delivery of fish by males to their mates ("courtship feeding") occurred prior-to, during, and following the egg-laying period. Following the laying of the second egg, courtship feeding rates declined significantly. There was a significant, positive correlation bebween courtship feeding rates and subsequent chick feeding rates by males. The incubation rates of females were significantly higher than those of males, especially during the first ten days of incubation. Territorial attendance rates during the incubation stage were similar for males and females. During the chick stage, territorial attendance rates of females were significantly higher than those of males. The size of fish fed to chicks by males increased as the chicks grew older and chick feeding rates of males were approximately three times higher than female rates. Based on these quantitative differences in parental care activities, the cumulative parental time investment by the two sexes was very similar. However, the energetic investment by males was likely greater than that by females, since male parental contributions (e.g. courtship feeding and chick feeding) often entailed extensive foraging behaviour.
    • Partial characterization of the cloned dihydrofolate reductase gene of Saccharomyces cerevisiae

      Nagel, Michael G.; Department of Biological Sciences (Brock University, 1985-07-09)
      The cloned dihydrofolate reductase gene of Saccharomyces cerevisiae (DFR 1) is expressed in Escherichia coli. Bacterial strain JF1754 transformed with plasmids containing DFR 1 is at least 5X more resistant to inhibition by the folate antagonist trimethoprim. Expression of yeast DFR 1 in E. coli suggests it is likely that the gene lacks intervening sequences. The 1.8 kbp DNA fragment encoding yeast dhfr activity probably has its own promotor, as the gene is expressed in both orientations in E. coli. Expression of the yeast dhfr gene cloned into M13 viral vectors allowed positive selection of DFR 1 - M13 bacterial transfectants in medium supplemented with trimethoprim. A series of nested deletions generated by nuclease Bal 31 digestion and by restriction endonuclease cleavage of plasmids containing DFR 1 physically mapped the gene to a 930 bp region between the Pst 1 and Sal 1 cut sites. This is consistent with the 21,000 molecular weight attributed to yeast dhfr in previous reports. From preliminary DNA sequence analysis of the dhfr DNA fragment the 3' terminus of DFR 1 was assigned to a position 27 nucleotides from the Eco Rl cut site on the Bam Hi - Eco Rl DNA segment. Several putative yeast transcription termination consensus sequences were identified 3' to the opal stop codon. DFR 1 is expressed in yeast and it confers resistance to the antifolate methotrexate when the gene is present in 2 - 10 copies per cell. Plasmid-dependent resistance to methotrexate is also observed in a rad 6 background although the effect is somewhat less than that conferred to wild-type or rad 18 cells. Integration of DFR 1 into the yeast genome showed an intermediate sensitivity to folate antagonists. This may suggest a gene dosage effect. No change in petite induction in these yeast strains was observed in transformed cells containing yeast dhfr plasmids. The sensitivity of rad 6 , rad 18 and wild-type cell populations to trimethoprim were unaffected by the presence of DFR 1 in transformants. Moreover, trimethoprim did not induce petites in any strain tested, which normally results if dhfr is inhibited by other antifolates such as methotrexate. This may suggest that the dhfr enzyme is not the only possible target of trimethoprim in yeast. rad 6 mutants showed a very low level of spontaneous petite formation. Methotrexate failed to induce respiratory deficient mutants in this strain which suggested that rad 6 might be an obligate grande. However, ethidium bromide induced petites to a level approximately 50% of that exhibited by wild-type and rad 18 strains.
    • Permissivity of human HeLa cells to bovine adenovirus type 2 (BaV2 infection

      Dobric, Nikolina.; Department of Biological Sciences (Brock University, 1998-07-09)
      Infection of hUlnan cells by bovine adenovirlls type 2 (BAV2) is abortive. To obtain a better understanding of this pllenomel1011, and in particular to identify Wllich steps in the viral replicative cycles are altered dllring this virlls-host cells interaction, we have llndertaken a detailed study of BAV2 infections of the nonpennissive hUlnan IIeLa cells. Using autoradiography and 3H-thymidine-labeled vvhole virus particles for infection of HeLa cells, vve determined that viral attachluent appears normal. Furthermore, Southern analysis revealed that internalization and transport to the nuclells occurs in BAV2 infected HeLa cells. To investigate viral DNi\ synthesis, infectivity assays involving hydroxyllrea, a viral DN-A synthesis inhibitor, were carried out. The results revealed that Bft:LV2 DNA synthesis does not occur in HeLa cells. Fllrtller investigations into viral early gene expression by northern blotting analyses indicated that HeLa cells fail to support expression of EIA. This suggested that abortive infection by BAV2 could be attributed to faiiure of EIA to express. To test the possibility that the failure to express ElA was due to the inability of the host cell to recognize the E lA prOlTIoter, ,ve carried out transient expression transfection experiments using plaslnids \vith the bacterial lacZ linder the control of either BAV2 or i\d5 EIA promoter. X-gal histochelIlical assays sho\ved expression of lacZ from the Ad5 ElA prOlnoter but no expression of lacZ [rOln the BAV2 EIA prOlTIoter. This further suggests that the abortive infection b:y BAV2 could be attributed to failure of EIA to express dlle to a nonfllnctional prOlTIoter in hlunan cells. Thus we speClllated that abortive infection of HeLa cells by adenoviruses may be averted by providing EtA functions in trans. To demonstrate this, we coinfected HeLa cells with Ad5 and BAV2, reasoning that Ad5 could cOlnpensate for EIA deficiency in BAV2. OUf results showed that BAV2 DNA synthesis was indeed Sllpported in HeLa cells coinfected with Ad5dlE3 as revealed by Southern analysis. In contrast, coinfection of HeLa cells \vith BAV2 and Ad5dlElE3 mutallt did not support BLt\V2 DNA synthesis. Interestingly, BAV2 failed to replicate in 293 cells which are constitlltively expressing the El genes. This could ilnply that El is necessary but not sufficient to avert the failllre ofBAV2 to undergo productive infection ofhulnan cells.
    • Perturbing the impulse activity of a single identified neuron augments the formation of long-term memory in a molluscan semi-intact preparation /

      Lowe, Michael R.; Department of Biological Sciences (Brock University, 2004-06-29)
      The aim of this study was to investigate the neural correlates of operant conditioning in a semi-intact preparation of the pond snail, Lymnaea stagnalis. Lymnaea learns, via operant conditioning, to reduce its aerial respiratory behaviour in response to an aversive tactile stimulus to its open pneumostome. This thesis demonstrates the successful conditioning of na'ive semiintact preparations to show learning in the dish. Furthermore, these conditioned preparations show long-term memory that persists for at least 18 hours. As the neurons that generate this behaviour have been previously identified I can, for the first time, monitor neural activity during both learning and long-term memory consolidation in the same preparation. In particular, I record from the respiratory neuron Right Pedal Dorsal 1 (RPeD 1) which is part of the respiratory central pattern generator. In this study, I demonstrate that preventing RPeDl impulse activity between training sessions reduces the number of sessions needed to produce long-term memory in the present semi-intact preparation.
    • Pharmacological analysis of 50 kHz vocalizations in the male rat

      Woodford, Timothy; Department of Biological Sciences
      The production of 50 kHz ultrasonic vocalizations in rats has been associated with both positive social interactions and appetitive behavioural situations. Furthermore, there is significant evidence showing that these vocalizations are controlled by the meso-limbic dopamine system. The purpose of this study was to perform a pharmacological analysis of 50 kHz calls by using dopamine and two dopamine agonists amphetamine and apomorphine, to induce calls. The acoustic parameters of the different call types were compared across each agonist. All three agonists were able to significantly induce more 50 kHz vocalizations compared to the vehicle control. Furthermore, calls elicited by apomorphine had a significantly higher bandwidth compared to those elicited by dopamine and amphetamine. All three agonists also had significantly different pharmacokinetic properties. These observations suggest that the D2 receptor sub-type is involved in the length of call bandwidths.
    • Photogrammetric Exploration of Demographic Change in Juvenile Rhizocarpon geographicum Thalli

      Bukovics, Theresa; Department of Biological Sciences
      Lichens, although ubiquitous and morphologically diverse, are one group of organisms that remains under-represented in life history and demographic studies, especially in early years. A photographic time series and image analysis were used to identify, track, and quantify morphological and demographic changes of the smallest, observable Rhizocarpon geographicum cohorts over a four-year period (2009 to 2013) at Illecillewaet Glacier, BC. Two objectives were examined: (a) quantify mortality, recruitment, and survivorship, and (b) examine trends correlated with thallus survival and visual, morphological changes in areole formation. Two general predictions were tested: (a) thallus survivorship is positively correlated with average areole area, and (b) there is a linear increase in the number of areoles per year per thallus. Results revealed age-specific mortality was highly variable among different years (8 to 44%), and recruitment occurred in every year. Thallus coalescence was found in every cohort and gradually increased over time. Results also showed that thalli that coalesced had a higher survivorship than solitary thalli. Survivorship was independent of areole area, and areole accumulation did not increase linearly in the number of areoles per year. Overall, this study demonstrates the use of a photographic time series to track and examine demographic patterns and quantify visual changes in morphology in small, newly found R. geographicum thalli. This thesis marked the first attempt at creating an age-specific cohort life table (up to 4 years) and tracking age-specific changes in morphology and survival in R. geographicum thalli.
    • The Physiological and Behavioural Consequences of Reduced Scalation in Captive-bred Phenotypes of the Bearded Dragon (Pogona vitticeps Ahl 1926)

      Sakich, Nicholas; Department of Biological Sciences
      Lepidosaurs as a group are known for their tough, scaled integument and low rates of evaporative water loss. Whether or not there is a causal relationship between the two has been a contentious issue. There also remains the question of whether the lepidosaur scale forms a barrier to ultraviolet (UV) light. Thirdly, there is evidence to suggest that rate of evaporative water loss influences behavioural thermoregulation in lepidosaurs. Lepidosaurs with higher rates of evaporative water loss should be expected to choose cooler temperatures than lepidosaurs with lower rates of evaporative water loss in order to reduce water loss. To investigate these ideas, I used three captive-occurring phenotypes of the bearded dragon (Pogona vitticeps Ahl 1926): Wild Type, animals exhibiting scales of reduced prominence (“Leatherback”), and scaleless animals (“Silkback”). I a priori expected that Silkbacks would have the highest rates of evaporative water loss, the lowest thermal preferences, and the lowest UV light intensity preferences. By the same token, I expected Wild Types to be at the opposite end of the spectrum from Silkbacks for each of these measurements, and I expected Leatherbacks to be intermediate between the two. I used respirometry to measure the animals’ rates of evaporative water loss, a thermal gradient to measure their thermal preferences, and a UV light intensity gradient to measure their UV light intensity preferences. Silkbacks on average lost water at about twice the rate that Wild Types did, with Leatherbacks being intermediate in their water loss rates. The three phenotypes did not visibly differ in their thermal preference. Silkbacks had lower UV light intensity preferences than either Leatherbacks or Wild Types. These results suggest that the lepidosaur scale is indeed a barrier to evaporative water loss and suggest that it is also a barrier to UV light. However, the lack of obvious difference in thermal preference suggests that thermal preference in bearded dragons is not plastic enough to respond to a phenotype that increases the animal’s rate of evaporative water loss. In addition to answering basic questions about lepidosaur biology, my data have relevance to the fields of animal welfare and conservation.
    • Phytoplankton species diversity of 27 lakes and ponds of Costa Rica (Central America)

      Umana Villalobos, Gerardo.; Department of Biological Sciences (Brock University, 1985-10-02)
      The literature on species diversity of phytoplankton of tropical lakes is scarce, and for the main part comes from studies of the big lakes in Africa, or deep lakes in South America, leaving a gap in the information about small shallow tropical lakes. In the present work the phytoplankton species composition and diversity of 27 shallow lakes and ponds in Costa Rica (Central America) was studied. The species composition was found to agree with other studies of tropical lakes, with a dominance of Chlorophyta, Cyanophyta, or in some cases Bacillariophyta or Euglenophyta; and a general paucity of Chrysophyta and Cryptophyta. Species richness varied considerably among the lakes, and tended to decrease with an increase in lake elevation. A low evenness in the species abundances was found, with one or more species outnumbering the rest by several orders of magnitude. Individual species abundances and species composition was found to vary with time in Rio Cuarto Lake, a meromictic lake situated in a region with low seasonal change in precipitation. In comparison with the phytoplankton of temperate lakes, the phytoplankton of the tropical lakes studied tended to have a lower evenness of species abundances, although species richness may be similar to temperate figures in some cases. Diversity indices sensitive to changes in the abundance of rare species tend to be higher in the tropical lakes studied; diversity indices sensitive to changes in the numbers of abundant species tend to be similar between the temperate and tropical lakes examined.