Analysis of the feeding behaviour of the mosquito Culex pipiens L. (Diptera: Culicidae) in relation to West Nile virus

The goal ofthis literature review is to inform the reader on several aspects of West Nile Virus (WNV) transmission by its mosquito vector, Culex pipiens and to elucidate how Cx. pipiens and WNV are intertwined. The first few sections of the literature review describe the life cycle and blood feeding behaviours ofmosquitoes so that baseline data ofmosquito biology are established. In addition to explaining how and why a mosquito blood feeds, the section on "Blood Meal Analysis" describes the different methods for determining the vertebrate source of mosquito blood meals and a brief history of these testing methods. Since this thesis looks at the feeding behaviour of Cx. pipiens, it is important to know how to determine what they are feeding upon. Discussion on other mosquito-borne diseases related to WNV gives a broader perspective to the thesis, and examines other diseases that have occurred in Ontario in the past. This is followed by background information on WNV and theories on how this virus came to North America and how it relates to Cx. pipiens. The final sections discuss Cx. pipiens and give background information to how this species of mosquito exists and behaves within North America.

Effects of reflection and social isolation on crayfish behaviour /

Visual stimuli and socialization influence exploratory behaviour in crayfish. The predominant components of spontaneous exploratory behaviour were determined by observing the activity of solitary adult crayfish (Procambarus clarkii) in a glass aquarium containing fresh water and no objects. Five distinct behaviours were observed: rearing up (climbing on the wall), turning around, cornering (facing the comer), backward walking, and crossing (crossing the midline of the aquarium). The frequency of rearing up, cornering and turning around decreased when reflection from the glass wall was blocked with black cardboard, black paint or non-reflective transparent plastic. In a tank containing mirrors on one side and non-reflective plastic on the other, crayfish cornered, reared up, and turned around more in front of the mirrors. Socialization was necessary for crayfish to respond to the reflection. Crayfish that were housed in pairs for two weeks exhibited more rearing up, turning around and cornering in front of the mirrors than in the non-reflective side. Crayfish isolated for two weeks did not show these differences. Socialized crayfish also exhibited more rearing up, turning around and cornering than did isolated crayfish. Thus, crayfish respond to visual stimuli provided by a glass tank, but the responds depends on socialization.

The efficacy of anti-predator behaviour in the wood fog tadpole (Rana sylvatica) /

Activity has been suggested as an important behaviour that is tightly linked with predator avoidance in tadpoles. In this thesis I examine predator-prey relationships using wood frog tadpoles {Rana sylvaticd) as prey and dragonfly larvae {AnaxJunius) and backswimmers {Notonecta undulatd) as predators. I explore the role of prey activity in predator attack rates, prey response to single and multiple predator introductions, and prey survivorship. The data suggest that Anax is the more successful predator, able to capture both active and inactive tadpoles. In contrast, Notonecta strike at inactive prey less frequently and are seldom successftil when they do. A mesocosm study revealed that the presence of any predator resulted in reduced activity level of tadpoles. Each predator species alone had similar effects on tadpole activity, as did the combined predator treatment. Tadpole survivorship, however, differed significantly among both predator treatments and prey populations. Tadpwles in the combined predator treatment had enhanced risk; survivorship was lower than that expected if the two predators had additive effects. Differences in survivorship among wood frog populations showed that tadpoles from a lake habitat had the lowest survivorship, those from a shallow pond habitat had an intermediate survivorship, and tadpoles from a marsh habitat had the highest survivorship. The frequency of interactions with predators in the native habitat may be driving the population differences observed. In conclusion, results from this study show that complex interactions exist between predators, prey, and the environment, with activity playing a key role in the survival of tadpoles.

Effect of citrate on the redox properties of cytochrome C and its kinetic and binding behaviour with cytochrome oxidase /

Increasing citrate concentration, at constant ionic strength (30 mM) decreases the rate of cytochrome ~ reduction by ascorbate. This effect is also seen at both high (600 mM) and low (19 mM) ionic strengths, and the Kapp for citrate increases with increasing ionic strength. Citrate binds d both ferri -and ferrocytochrome ~, but with a lower affinity for the latter form (Kox . .red d = 2 mM, Kd = 8 mM) as shown by an equilibrium assay with N,N,N',N', Tetramethyl E- phenylenediamine. The reaction of ferricytochrome ~with cyanide is also altered in the presence of citrate: citrate increases the K~PP for cyanide. Column chromatography of cytochrome ~-cytochrome oxidase mixtures shows citrate increases the dissociation constant of the complex. These results are confirmed in kinetic assays for the "loose"site (Km = 20 pM) only. The effect of increasing citrate observable at the "tight" site (Km = 0.25 pM) is on the turnover number and not on the K . These results suggest a mechanism m where anion binding to cytochrome £ at the tight site affects the equilibrium between two forms of cytochrome c bound cytochrome oxidase: an active and an inactive one.

Host-parasite relationships in mycoparasite /

A mycoparasite, Piptocephalis virginiana ^ shows a resemblance to fungal parasites of higher plants in the fine structure of hyphae and haustoria. The morphology and fine structure of host and parasitic fungi have been described. The mode of penetration of the host cell, Choanephora cucurbitarum , probably involves mechanical forces. Although the presence of cell wall degrading enzyme was not detected by conventional techniques, its role in penetration can't be ruled out. A collar around the haustorial neck is formed as an extension of the host cell wall. No papilla was detected although appressorixim was seen during penetration. The young haustorium is enclosed in highly invaginating plasmalemma of the host cell and n\imerous cisternae of endoplasmic reticulum. Appearance of an electron—dense sheath around the mature haustorium seems to coincide with the disappearance of cisternae of endoplasmic reticulum from the host cystoplasm in the vicinity of the haustorium. The role of host cytoplasm particularly of endoplasmic reticulum in the development of the sheath is discussed. Extensive accumulation of spherosomes-like bodies, containing lipids, is found in haustorium, parasite and host hypha. Electron microscope revealed the parasiticculture spore has more lipid content than the axenic culture spore of P. virginiana . The biochemical and cytochemical tests also support these results. The mature spore of C. cucurbitarum possesses a thick three-layered cell wall, different from the hyphal wall. Its germination is accompanied by the formation of an elastic thin inner layer which surrounds the emerging germ tube and the growing hypha. High resolution autoradiography showed that H N-acetyl-glucosamine , a precursor of chitin, was incorporated preferentially in the thin inner layer of the spore wall and also in the cell wall of the growing hypha. When the label was fed to the infected cells, at different intervals after inoculation, grains were observed on the sheath which developed around the haustorium of P. virginiana , 30 hours after inoculation. The significance of these results in relation to the origin and composition of the sheath is discussed.

Host-parasite relations in a mycoparasitic system : alterations in membrane permeability and lipid composition in Choanephora cucurbitarum infected by Piptocephalis virginiana /

examined in Choanephora cucurbita rum during the early stages of infection by Piptocephalis virginiana » There was a small but consistent increase in the leakage of electrolytes, amino acids and sugars as a result of infection. These low levels of differential leakage in infected tissues are explained on the basis of the nature of this obligate, biotrophic, mycoparasitic system. Quantitative analysis of the twenty six amino acids and amino compounds detected in the leacheates — showed similar profiles in infected and control host and no new species of amino acids or amino compounds were detected in either infected or control host leacheates. Comparatively high amounts of aspartic acid, glutamic acid and alanine were found in the leacheates of host and infected host . Analyses of the sugars comprising the leacheates of infected and control host showed the presence of eight sugars, among which glucose was found in significant amounts (50-53%) ' The nutritional implication of this preferential leakage is discussed. No significant difference was observed in the leacheates of infected host sugar profiles compared with that of the control host. Profiles of the internal pool sugars of infected and control host did not reflect that obtained from the leacheate data, perhaps owing to leakage of sugars in a selective manner . Membrane lipid analyses yielded higher levels of lipid in infected host compared with the control, both at the 24 h and 36 h analyses. In addition, preliminary investigations of phosphorous-32 incorporation and turnover in phospholipids showed higher levels of 32p incorporation and turnover in infected host compared with the control. No apparent difference was noted in the profiles of the neutral lipid classes and the polar lipid classes of the membrane lipids as determined by one and two dimensional thin-layer chromatography respectively. However, a small but consistently higher degree of unsaturation was detected in the fatty acids of infected tissue compared with the control. Also, '^''-^^''^^'-'-^'^^c acid, a polyunsaturated fatty acid previously reported to show a direct correlation during the early stages of infection and the degree of parasitism of P. virginiana on C. cucurbitarum , was found in higher amounts in infected host membrane lipids compared with that of the control host. The implications of these membrane lipid alterations are discussed with particular reference to the small but consistently higher leakage of electrolytes, amino acids and sugars observed during infection in this study.

Anxiety and learning / response behaviour in the context of intellectual problem solving by young children

Twenty-eight grade four students were ca.tegorized as either high or low anxious subjects as per Gillis' Child Anxiety Scale (a self-report general measure). In determining impulsivity in their response tendencies, via Kagan's Ma.tching Familiar Figures Test, a significant difference between the two groups was not found to exist. Training procedures (verbal labelling plus rehearsal strategies) were introduced in modification of their learning behaviour on a visual sequential memory task. Significantly more reflective memory recall behaviour was noted by both groups as a result. Furthermore, transfer of the reflective quality of this learning strategy produced significantly less impulsive response behaviour for high and low anxious subjects with respect to response latency and for low anxious subjects with respect to response accuracy.

A social-cognitive model of monitoring and its relationship to behaviour in a naturalistic observation : development and individual differences

years 8 months) and 24 older (M == 7 years 4 months) children. A Monitoring Process Model (MPM) was developed and tested in order to ascertain at which component process ofthe MPM age differences would emerge. The MPM had four components: (1) assessment; (2) evaluation; (3) planning; and (4) behavioural control. The MPM was assessed directly using a referential communication task in which the children were asked to make a series of five Lego buildings (a baseline condition and one building for each MPM component). Children listened to instructions from one experimenter while a second experimenter in the room (a confederate) intetjected varying levels ofverbal feedback in order to assist the children and control the component ofthe MPM. This design allowed us to determine at which "stage" ofprocessing children would most likely have difficulty monitoring themselves in this social-cognitive task. Developmental differences were obselVed for the evaluation, planning and behavioural control components suggesting that older children were able to be more successful with the more explicit metacomponents. Interestingly, however, there was no age difference in terms ofLego task success in the baseline condition suggesting that without the intelVention ofthe confederate younger children monitored the task about as well as older children. This pattern ofresults indicates that the younger children were disrupted by the feedback rather than helped. On the other hand, the older children were able to incorporate the feedback offered by the confederate into a plan ofaction. Another aim ofthis study was to assess similar processing components to those investigated by the MPM Lego task in a more naturalistic observation. Together the use ofthe Lego Task ( a social cognitive task) and the naturalistic social interaction allowed for the appraisal of cross-domain continuities and discontinuities in monitoring behaviours. In this vein, analyses were undertaken in order to ascertain whether or not successful performance in the MPM Lego Task would predict cross-domain competence in the more naturalistic social interchange. Indeed, success in the two latter components ofthe MPM (planning and behavioural control) was related to overall competence in the naturalistic task. However, this cross-domain prediction was not evident for all levels ofthe naturalistic interchange suggesting that the nature ofthe feedback a child receives is an important determinant ofresponse competency. Individual difference measures reflecting the children's general cognitive capacity (Working Memory and Digit Span) and verbal ability (vocabulary) were also taken in an effort to account for more variance in the prediction oftask success. However, these individual difference measures did not serve to enhance the prediction oftask performance in either the Lego Task or the naturalistic task. Similarly, parental responses to questionnaires pertaining to their child's temperament and social experience also failed to increase prediction oftask performance. On-line measures ofthe children's engagement, positive affect and anxiety also failed to predict competence ratings.

Behavioural characteristics in phylogenetics : a case study using black fly (Diptera: Simuliidae) cocoon spinning behaviour

1-1 is torically, the predominan t method of reconstructing phylogenies has been through the use of morphological characters. There are new techniques now gaining acceptance, including molecular techniques al1d chromosomal information. Altl10ugh the study of behaviour has been used in a comparative framework, these analyses have, historically, been based on intuition. Hennig (1966) devised a neV\' method of reconstructing phylogenies which provided a 110ncircular method for formulating, testing and refining phylogenies. Subsequent s)Tstematists had virtually abandoned ecological and beha\lioural data as primary indicators of phylogenetic relationships (Brooks and McLennan 1991). Therefore, in a modern cladistic framework (sensu Hennig) the analysis of behavioural traits remains underrepresented as a method of reconstructing phylogenies. This thesis will reconstruct the phylogeny for species of black flies (Diptera: Simuliidae), using two steps. The first step is to thoroughl)' understand and explain the cocoon spinning in black fly larvae. There have bee115 previous descriptions of cocoon spinning, but all were incomplete or erroneous. The advances in technology, including video recorders and VCRs, have allowed this behaviour to be analyzed in great detail in 20 different species. A complete description of the cocoon spinning of Simulium \littatum is given. This description will be used as a template for the other species observed. The description and understanding of cococ)n spinning was the first step in undertaking a phylogenetic analysis using this behaviour. The behaviour was then broken down and analyzed, revealing 23 characters, 3 either qualitative and quantitative in nature. These characters were assessed in a cladistic framework (sensu Hennig) and a phylogenetic tree was reconstructed with a e.I of 0.91 and an R.I. of 0.96. This phylogenetic tree closely resembles a previously established pllylogenetic tree produced from morphological and cytological information. The importance of this result is the indication that, contrary to some authors, behavioural characters, if used properly, can add very informative characters to a data set.

Effectiveness of life skills training on the total behaviour of six individuals in a job preparation program

In the first week of a Job preparation Program all twelve female members of the class were invited to be part of a study to determine the effect of Life Skills training on their behaviour over the twelve-week period of the program. Six females volunteered and each was interviewed four times during the Job preparation Program and once after the program ended. The interviews focused on three areas of skill deficiency addressed in Life Skills lessons: their knowledge about themselves and attitude towards themselves; their interpersonal relationships; and their problem-solving ability. The participants' comments over the sixteen-week period of the interviews were used to decide if the total behaviour of the participants, (i.e., what they did, thought, and felt) changed so that each became more effective in satisfying her needs. The study suggested that the total behaviour of three of the six women changed so that they became more effective in satisfying their needs. The fourth female's total behaviour changed in only two of the three areas focused on in the interviews, and the total behaviour of the fifth and sixth females showed no change.

Individual differences and factors affecting male behaviour in field crickets

Individual differences in male sexual behav~our and the factors influencing calling behaviour were studied in the field crickets Gryllus 2 integer and Q. veletis. In a large (13m) outdoor arena individually numbered adult male ~~ integer started calling at three to five days of age but thereafter the age of individual G. integer males did not affect nightly calling duration. Calling also did not correlate with individual weight. In this study individual male calling was continuously distributed from 0 hrs. per night to 3.5 hrs. per night, on average. A temporal effect on the number of G. integer males calling was observed. The number of males calling through the night was uniform, but a sharp increase in the number calling was observed in the early morning. No difference in calling times was observed between the night and dawn callers. AlsC)' males calling at dawn usually didnotc'all during the preceeding night. Calling and reproductive success in 1979 demonstrated a negative logarithmic relationship while in the 1980(initial) population a negative linear relationship was observed. No relationship was seen in the 1980 high density population. The ratio of non-callers to callers also affected the mating of individuals in the 1979 and1980(initial) densities:-non~callers (males calling .5 hrs. per night, on average, or less) obtained more females when the population contained a high number of callers, this being a negative logarithmic relationship to, No such relationship was observed in the 1980 high density population. Individual displacement varied nightly and was not correlated to amount of calling or reproductive success of individual G. integer males. G. integer males were displa~ed more when in a higher density in the outdoor arena Male G. integer and G. veletis behaviours were also observed in an indoor arena at different densities and, in G. veletis, with respect to female presence. When females were present in the arena, in G. veletis, male calling was reduced. Males of both species called less, on average, when in ~ higher density, than when they were in a lower density. Male displacement of both species increased on average when in a higher density as compared to displacement in a lower density. Aggression was measured by aggressive call-ing and fighting and was studied in regards to density.G. integer demonstrated less aggression in all but one comparison at higher density. No difference was observed in the ratio of aggressive calling to f.ighting comparison in G. integer. G. veletis demonstrated mixed results. No difference in aggression between densities was observed in comparisons. Less.aggression did occur in higher densities when comparisons invol.ved fighting behaviour. Male behaviour represents a competitive strategy against ot~er males, strategy being defined as a genetic (in part) alternative to other strategies. In this sense, the factors of time, density, male-male aggression, and female presence are conditions demonstrated to affect male behaviour in G. integer and G. veletis. Individual male differences and other considerations suggest that alternative male behaviours are represented by at least two conditional strategies. This possibility, and the transient 'or stable nature of genetic polymorphisms in field cricket behaviour are considered.

Involvement of fimbriae in host-mycoparasite recognition

Extracellular, non-flagellar appendages, termed fimbriae are widespread among fungi. Fungal fimbriae range in diameter from 6-10 nm and exhibit lengths of up to 30 ~m. Fungal fimbriae have been implicated in several functions: adhesion, conjugation and flocculation. A possible role of fimbriae in host-mycoparasite interactions was the focus of this study . Using electron microscopy, fimbriae were observed on the surfaces of Mortiere lla cande labrum, Mortie re lla pusi lla and Phascolomyces articulosus with diameter means of 9.1±0.4 nm, 9.4±0.5 nm and 8.6±0.6 nm, respectively, and lengths of up to 25 ~m. Fimbriae were not observed on the surface of the mycoparasite, Piptocephalis virginiana. Polyclonal antiserum (AU) prepared against the fimbrial protein of Ustilago violacea cross-reacted with 60 and 57 kDa M. candelabrum proteins. In addition, AU cross-reacted with 64 kDa proteins from both M. pusilla and P. articulosus. The proteins that cross-reacted with AU were electroeluted from polyacrylamide gels and were shown to subsequently form fibrils. The diameter means for the electroeluted fibrils were: for M. candelabrum 9.7±0.3 nm, M. pusilla 8.4±0.6 nm and P articulosus 9.2±0.5 nm. Finally, to ascertain the role of fimbriae in host-mycoparasite interactions, AU was incubated with P. virginiana and M. pusilla (mycoparasite/susceptible host) and with P. virginiana and P . articulosus (mycoparasite/ resistant host). It was observed that AU decreased significantly the level of contact between P. virginiana and M. pusilla and between P. virginiana and P. articulosus in comparison to prelmmune serum treatments. Thus, it was proposed that fimbriae might play recognition and attachment roles in early events of mycoparasitism.

Isolation and partial characterization of host cell wall surface glycoproteins: their involvement in agglutination, attachment and appressorium formation by piptocephalis virginiana

Cell surface proteins obtained by alkaline extraction from isolated cell walls of Mortierella pusilla and M. candelabrum, host and nonhost, respectively, to the mycoparasite, Piptocephalis virginiana, were tested for their ability to agglutinate mycoparasite spores. The host cell wall protein extract had a high agglutinating activity (788 a.u. mg- t ) as compared with the nonhost extract (21 a.li. mg- t ). SDS-polyacrylamide gel electrophoresis of the cell wall proteins revealed four protein bands, a, b, c, and d (Mr 117, 100, 85 and 64 kd, respectively) at the host surface, but not at the nonhost surface, except for the faint band c. Deletion of proteins b or c from the host cell wall protein extract significantly reduced its agglutinating activity. Proteins band c, obtained as purified preparations by a series of procedures, were shown to be two glycoproteins. Carbohydrate analysis by gas chromatography demonstrated that glucose and Nacetylglucosamine were the major carbohydrate components of the glycoproteins. It was further shown that the agglutinating activity of the pure preparation containing both band c was 500-850 times that of the single glycoproteins, suggesting the involvement of both glycoproteins in agglutination. The results suggest that the glycoproteins band c are the two subunits of agglutinin present at the host cell surface. The two glycoproteins band c purified from the host cell wall protein extract were further examined after various treatments for their possible role in agglutination, attachment and appressorium formation by the mycoparasite. Results obtained by agglutination and attachment tests showed: (1) the two glycoprotein-s are not only an agglutinin responsible for the mycoparasite spore agglutination, but may also serve as a receptor for the specific recognition, attachment and appressorium formation by the mycoparasite; (2) treatment of the rnycoparasite spores with various sugars revealed that arabinose, glucose and N-acetylglucosamine inhibited the agglutination and attachment activity of the glycoproteins, however, the relative percentage of appressorium formation was not affected by the above sugars; (3) the two glycoproteins are relatively stable with respect to their agglutinin and receptor functions. The present results suggest that the agglutination and attachment may be mediated directly by certain sugars present at the host and mycoparasite cell surfaces while the appressorlum formation may be the response of complementary combinations of both sugar and protein, the two parts of the glycoproteins at the interacting surfaces of two fungi.

Isolation and partial characterization of a complementary protein from the mycoparasite, Piptocephalis virginiana, which specifically binds to two glycoproteins b and c of the host cell surface

Presence of surface glycoprotein in Piptocephalis virginiana that recognizes the host glycoproteins band c, reported earlier from our laboratory, was detected by immunofluorescence microscopy. Germinated spores of P. virginiana treated with Mortierella pusilla cell wall protein extract, primary antibodies prepared against glycoproteins band c and FITC-goat anti-rabbit IgG conjugate showed fluorescence. This indicated that on the surfaces of the biotrophic mycoparasite P. virginiana , there might be a complementary molecule which recognizes the glycoproteins band c from M. pusilla. Immunobinding analysis identified a glycoprotein of Mr 100 kDa from the mycoparasite which binds with the host glycoproteins band c, separately as well as collectively. Purification of this glycoprotein was achieved by (i) 60% ammonium sulfate precipitation, (ii) followed by heat treatment, and (iii) Sephadex G-IOO gel filtration. The glycoprotein was isolated by preparative polyacrylamide gel electrophoresis by cutting and elution. The purity of the protein ·was ascertained by SDS-PAGE and Western blot analysis. Positive reaction to periodic acid-Schiff reagent revealed the glycoprotein nature of this 100 kDa protein. Mannose was identified as a major sugar component of this glycoprotein by using a BoehringerMannheim Glycan Differentiation Kit. Electrophoretically purified glycoprotein was used to raIse polyclonal antibody in rabbit. The specificity of the antibody was determined by dot-immunobinding test and western-blot analysis. Immunofluorescence mIcroscopy revealed surface localization of the protein on the germ tube of Piptocephalis virginiana. Fluorescence was also observed at the surfaceJ of the germinated spores and hyphae of the host, M. pusilla after treatment with complementary protein from P. virginiana, primary antibody prepared against the complementary protein and FITC-goat anti-rabbit IgG conjugate.

Light and electron microscope studies of host-parasite relations in a mycoparasite

Light microscope studies of the mycoparasite Piptocephalis virginiana revealed that the cylindrical spores of the parasite became spherical upon germination and produced 1-4 germ tubes. Generally t"l.vO germ tubes were produced by each spore. When this parasite was inoculated on its potential hosts, Choanephora cucurbitarum and Phascolomyces articulosus, the germ tube nearest to the host hypha continued to grow and made contact with the host hypha. The tip of the parasite's germ tube became swollen to form a distinct appressorium. Up to this stage the behavior of the parasite was similar regardless of the nature of the host. In the compatible host-parasite combination, the parasite penetrated the host, established a nutritional relationship and continued to grow to cover the host completely with its buff colored spores in 3-4 days. In the incompatible host-parasite combination, the parasite penetrated the host but its further advance was arrested. As a result of failure to establish a nutritional relationship with the resistant host, the parasite made further attempts to penetrate the host at different sites producing multiple infections. In the absence of nutrition the parasite weakened and the host outgrew the parasite completely. In the presence of a non-host species, Linderina pennispora the parasite continued to grow across the non-host 1).yp_hae vlithout establishing an initial contact. Germination studies showed that the parasite germinated equally well in the presence of host and non-host species. Further electron microscope studies revealed that the host-parasite interaction between P. virginiana and its host, C. cucurbi tarum, was compatible when the host hyphae were young slender, with a thin cell wall of one layer. The parasite appeared to penetrate mechanically by pushing the host-cell wall inward. The host plasma membrane invaginated along the involuted cell wall. The older hyphae of C. cucurbitarum possessed two distinct layers of cell wall and-showed an incompatible interaction when challenged vlith the parasite. At the point of contact, the outer layer of the host-cell wall dissolved, probably by enzymatic digestion, and the inner layer became thickened and developed a papilla as a result of its response to the parasite. The haustoria of the parasite in the old hyphae were always surrounded by a thick, well developed sheath, whereas the haustoria of the same age in the young host mycelium were devoid of a sheath during early stages of infection. Instead, they were in direct contact with the host protoplast. The incompatible interaction between a resistant host, P. articulosus and the parasite showed similar results as with the old hyphae of C. cucurbitarum. The cell wall of P. articulosus appeared thick-with two or more layers even in the 18-22 h-old hyphae. No contact or interaction was established between the parasite and the non-host L. pennispora. The role of cell wall in the resistance mechanism is discussed.