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Biomedical subjects

Boris R Krasnov

Publications and source records attributed to Boris R Krasnov.

At least 19 recordsLinked to original sources

Locomotor response to light and surface angle in three species of desert fleas.

We studied the relocation of newly emerged and fed individuals of three species of desert fleas (Xenopsylla conformis, Xenopsylla ramesis, and Parapulex chephrenis) in response to light and surface angle. We observed flea movements inside of either horizontal or tilted cardboard tubes with different light regime at their ends. Proportion of relocating X. conformis and X. ramesis was significantly higher than that of P. chephrenis. In this species only, adult individuals relocated more frequently than newly emerged individuals, and females relocated more frequently than males. In general, the majority of fleas moved toward light independently of its position in relation to the surface angle. Fleas moved toward light even if it was positioned at the lower end of a tube. When both ends of a tube were darkened, newly emerged Xenopsylla moved randomly toward the upper or lower end of a tube, whereas newly emerged P. chephrenis moved mainly toward the upper end of a tube. Adult P. chephrenis and X. conformis also moved mainly toward the upper end of a tube, whereas adult X. ramesis moved mainly toward the lower end. When both ends of a tube were lighted up, newly emerged females of all species, as well as newly emerged female X. ramesis, randomly relocated toward the upper or lower end of a tube. In contrast, newly emerged males and adults of both sexes of P. chephrenis and X. conformis as well as adult female X. ramesis moved mainly toward the upper end of a tube, whereas adult male X. ramesis moved mainly down. Results of this study suggest that light is a more important abiotic signal for flea orientation than surface angle, and there are species-specific differences in flea responses to light and angle stimuli. These differences are related to spatial ecology and behavior of fleas' main hosts as well as to fleas' environmental preferences.

Age Factors↗

Temporal dynamics of a T-cell mediated immune response in desert rodents.

Immunocompetence, the general capacity of an individual host to mount an immune response against pathogens, is commonly assessed by the response to a challenge of the immune system by injection of phytohaemagglutinin (PHA). The response to PHA is commonly considered a reliable estimate of the T-cell mediated immune response. We investigated the temporal pattern of the PHA response in 10 rodent species from the Negev desert, Israel. We hypothesized that the temporal dynamics of the PHA response would differ among species with different natural patterns of flea parasitism. We injected PHA subcutaneously in the footpad of each rodent and measured its PHA response 6, 24 and 48 h after injection. Rodent species showed two types of PHA response. One type was rapid and characteristic of rodents that had either species-poor flea assemblages, or that are rarely attacked by fleas. This response peaked approximately 6 h after PHA injection. The second type of response was delayed and was typical of rodents that have either species-rich flea assemblages or high abundance and prevalence of fleas or both. Their response to PHA peaked 24 h after injection. Furthermore, rodents that responded promptly had a lower maximum response than rodents with a delayed response. Our results suggest the occurrence of a trade-off between intensity and latency of the PHA response and, therefore, the necessity to account for the relationship between maximum PHA response and time after injection when making interspecific comparisons of immunocompetence.

Animals↗

Is abundance a species attribute? An example with haematophagous ectoparasites.

Population density is a fundamental property of a species and yet it varies among populations of the same species. The variation comes from the interplay between intrinsic features of a species that tend to produce repeatable density values across all populations of the same species and extrinsic environmental factors that differ among localities and thus tend to produce spatial variation in density. Is inter-population variation in density too large for density to be considered a true species character? We addressed this question using data on abundance (number of parasites per individual host, i.e. equivalent to density) of fleas ectoparasitic on small mammals. The data included samples of 548 flea populations, representing 145 flea species and obtained from 48 different geographical regions. Abundances of the same flea species on the same host species, but in different regions, were more similar to each other than expected by chance, and varied significantly among flea species, with 46% of the variation among samples accounted by differences between flea species. Thus, estimates of abundance are repeatable within the same flea species. The same repeatability was also observed, but to a lesser extent, across flea genera, tribes and subfamilies. Independently of the identity of the flea species, abundance values recorded on the same host species, or in the same geographical region, also showed significant statistical repeatability, though not nearly as strong as that associated with abundance values from the same flea species. There were also no strong indications that regional differences in abiotic variables were an important determinant of variation in abundance of a given flea species on a given host species. Abundance thus appears to be a true species trait in fleas, although it varies somewhat within bounds set by species-specific life history traits.

Animals↗

Ecological characteristics of flea species relate to their suitability as plague vectors.

The ability of vector-borne diseases to persist and spread is closely linked to the ecological characteristics of the vector species they use. Yet there have been no investigations of how species used as vectors by pathogens such as the plague bacterium differ from closely related species that are not used as vectors. The plague bacterium uses mammals as reservoir hosts and fleas as vectors. The ability of different fleas to serve as vectors is assumed to depend on how likely they are to experience gut blockage following bacterial multiplication; the blockage causes fleas to regurgitate blood into a wound and thus inject bacteria into new hosts. Beyond these physiological differences, it is unclear whether there exist fundamental ecological differences between fleas that are effective vectors and those that are not. Here, using a comparative analysis, we identify clear associations between the ability of flea species to transmit plague and their ecological characteristics. First, there is a positive relationship between the abundance of flea species on their hosts and their potential as vectors. Second, although the number of host species exploited by a flea is not associated with its potential as a vector, there is a negative relationship between the ability of fleas to transmit plague and the taxonomic diversity of their host spectrum. This suggests a correlation between some ecological characteristics of fleas and their ability to develop the plague blockage. The plague pathogen thus uses mainly abundant fleas specialized on a narrow taxonomic range of mammals, features that should maximize the persistence of the disease in the face of high flea mortality, and its transmission to suitable hosts only. This previously unrecognized pattern of vector use is of importance for the persistence and transmission of the disease.

Animals↗

Temporal variation in parasite infestation of a host individual: does a parasite-free host remain uninfested permanently?

Host individuals not infested by parasites at a given time are either permanently free from parasites or could be infested at other times. We studied temporal variation in the presence or absence of fleas (Siphonaptera) on individual rodents of two species (Gerbillus dasyurus and Acomys cahirinus) and questioned if and how an individual rodent can change its infestation status temporally. Change in infestation status by fleas over time was found in 45.5% of G. dasyurus and 35.9% of A. cahirinus. In both host species, the proportion of individuals that either changed or did not change their infestation status did not differ from the null expectation of 1:1. No difference between the proportions of infested at two consecutive captures and uninfested at two consecutive captures animals was found in A. cahirinus, whereas the proportion of G. dasyurus infested by fleas at two consecutive captures was significantly greater than that of uninfested at two consecutive captures individuals. In both host species, similar proportions of individuals changed their infestation status either from being infested to being uninfested or vice versa. Among individuals of both species that were initially infested by fleas, similar proportions changed and did not change their subsequent infestation status. The same was true for the uninfested at the first capture A. cahirinus. However, among initially uninfested G. dasyurus, the proportion of individuals that remained uninfested was significantly greater than that of the individuals newly infested by fleas. In A. cahirinus, the probability to change the infestation status did not depend on the initial status of an individual. In G. dasyurus, the probability of the initially uninfested individuals to be subsequently found still uninfested was higher than to be subsequently found harboring fleas, whereas initially infested individuals could be subsequently found either still infested or flea-free with equal probability.

Animals↗

Ectoparasites and age-dependent survival in a desert rodent.

Host age is one of the key factors in host-parasite relationships as it possibly affects infestation levels, parasite-induced mortality of a host, and parasite distribution among host individuals. We tested two alternative hypotheses about infestation pattern and survival under parasitism in relation to host age. The first hypothesis assumes that parasites are recruited faster than they die and, thus, suggests that adult hosts will show higher infestation levels than juveniles because the former have more time to accumulate parasites. The second hypothesis assumes that parasites die faster than they are recruited and, thus, suggests that adults will show lower infestation levels because of acquired immune response and/or the mortality of heavily infested juveniles and, thus, selection for less infested adults. As the negative effects of parasites on host are often intensity-dependent, we expected that the age-related differences in infestation may be translated to lower or higher survival under parasitism of adults, in the cases of the first and the second hypotheses, respectively. We manipulated ectoparasite numbers using insecticide and assessed the infestation pattern in adult and juvenile gerbils (Gerbillus andersoni) in the Negev Desert. We found only a partial support for age-dependent parasitism. No age-related differences in infestation and distribution among host individuals were found after adjusting the ectoparasite numbers to the host's surface area. However, age-related differences in survival under parasitism were revealed. The survival probability of parasitized juveniles decreased in about 48% compared to unparasitized hosts while the survival probability of adults was not affected by ectoparasites. Our results suggest that the effect of host age on host-parasite dynamics may not explicitly be determined by age-dependent differences in ectoparasite recruitment or mortality processes but may also be affected by other host-related and parasite-related traits.

Aging↗

Relationship between host abundance and parasite distribution: inferring regulating mechanisms from census data.

1. We studied the effect of host abundance on parasite abundance and prevalence using data on 57 associations of fleas (Siphonaptera) and their mammalian hosts from Slovakia. 2. We assumed that flea-induced host mortality could be inferred from the relationship between flea aggregation and flea abundance, whereas host-induced flea mortality could be inferred from the relationship between flea abundance or aggregation and host abundance. 3. Relationships between flea abundance or prevalence and host abundance were either negative (in 23 flea-host associations) or absent (in 34 flea-host associations). Negative relationships between flea abundance and host abundance were always accompanied by negative relationships between flea prevalence and host abundance. 4. The link between flea abundance/prevalence and host abundance was evaluated as the coefficient of determination of the respective regressions. Across flea-host associations, this link decreased with an increase in the degree of flea aggregation (measured as a parameter b of Taylor's power law). 5. Mean crowding of fleas decreased with an increase of host abundance in eight flea-host associations, being asymptotic in four of them. On the other hand, mean crowding of fleas increased with an increase in flea abundance in 49 flea-host associations, being asymptotic in 15 of them. 6. Results of this study suggest that different flea-host associations are governed by different regulating mechanisms, but different regulation mechanisms may act simultaneously within the same flea-host associations.

Animals↗

Are ectoparasite communities structured? Species co-occurrence, temporal variation and null models.

1. We studied temporal variation in the structure of flea communities on small mammalian hosts from eastern Slovakia using null models. We asked (a) whether flea co-occurrences in infracommunities (in the individual hosts) in different hosts as well as in the component communities (in the host species) demonstrate a non-random pattern; (b) whether this pattern is indicative of either positive or negative flea species interactions; (c) whether this pattern varies temporally; and (d) whether the expression of this pattern is related to population size of either fleas or hosts or both. 2. We constructed a presence/absence matrix of flea species for each temporal sample of a host species and calculated four metrics of co-occurrence, namely the C-score, the number of checkerboard species pairs, the number of species combinations and the variance ratio (V-ratio). Then we compared these metrics with the respective indices calculated for 5000 null matrices that were assembled randomly using two algorithms, namely fixed-fixed (FF) and fixed-equiprobable (FE). 3. Most co-occurrence metrics calculated for real data did not differ significantly from the metrics calculated for simulated matrices using the FF algorithm. However, the indices observed for 42 of 75 presence/absence matrices differed significantly from the null expectations for the FE models. Non-randomness was detected mainly by the C-score and V-ratio metrics. In all cases, the direction of non-randomness was the same, namely the aggregation, not competition, of flea species in host individuals and host species. 4. The inclusion or exclusion of the uninfested hosts in the FE models did not affect the results for individual host species. However, exclusion of the uninfested host species led to the acceptance of the null hypothesis for only six of 13 temporal samples of the component flea communities for which non-randomness was detected when the uninfested hosts were included in the analysis. 5. In most host species, the absolute values of the standardized size effect of both the C-score and V-ratio increased with an increase in host density and a concomitant decrease in flea abundance and prevalence. 6. Results of this study demonstrated that (a) flea assemblages on small mammalian hosts were structured at some times, whereas they appeared to be randomly assembled at other times; (b) whenever non-randomness of flea co-occurrences was detected, it suggested aggregation but never segregation of flea species in host individuals or populations; and (c) the expression of structure in flea assemblages depended on the level of density of both fleas and hosts.

Animals↗

Age-dependent flea (Siphonaptera) parasitism in rodents: a host's life history matters.

We studied age-dependent patterns of flea infestation in 7 species of rodents from Slovakia (Apodemus agrarius, A. flavicollis, A. sylvaticus, A. uralensis, Clethrionomys glareolus, Microtus arvalis, and M. subterraneus). We estimated the age of the host from its body mass and expected the host age-dependent pattern of flea abundance, the level of aggregation, and prevalence to be in agreement with theoretical predictions. We expected that the mean abundance and the level of aggregation of fleas would be lowest in hosts of smallest and largest size classes and highest in hosts of medium size classes, whereas pattern of variation of prevalence with host age would be either convex or asymptotic. In general, mean abundance and species richness of fleas increased with an increase in host age, although the pressure of flea parasitism in terms of number of fleas per unit host body surface decreased with host age. We found 2 clear patterns of the change in flea aggregation and prevalence with host age. The first pattern demonstrated a peak of flea aggregation and a trough of flea prevalence in animals of middle age classes (Apodemus species and C. glareolus). The second pattern was an increase of both flea aggregation and flea prevalence with host age (both Microtus species). Consequently, we did not find unequivocal evidence for the main role of either parasite-induced host mortality or acquired resistance in host age-dependent pattern of flea parasitism. Our results suggest that this pattern can be generated by various processes and is strongly affected by natural history parameters of a host species such as dispersal pattern, spatial distribution, and structure of shelters.

Age Factors↗

Relationships between local and regional species richness in flea communities of small mammalian hosts: saturation and spatial scale.

The number of species coexisting in a community may be regulated by local factors (e.g., competitive interactions), or by regional processes (e.g., dispersal from a regional species pool). The relative importance of local and regional processes can be inferred from the shape of the relationship between local and regional species richness. We investigated this relationship in communities of fleas parasitic on small mammals at two spatial scales: between the richness of fleas on individual hosts (infracommunities) and that of fleas on host populations (component communities), and between the richness of component communities and that of the entire regional species pool. We tested linearity (proportional sampling) versus curvilinearity with an asymptote (species saturation) by plotting "local" against "regional" species richness of fleas either among host species or within host species among populations. At the two spatial scales, we found consistent curvilinear relationships between species richness of the more "local" communities and richness of the more "regional" communities. This was true across all host species in the data set and for geographic subsets, even after controlling for the influence of sampling effort on estimates of species richness, and that of host phylogeny in interspecific analyses. We also tested for density compensation in species-poor communities. There was no strong evidence for density compensation at the infracommunity level, although its existence at the component community level appeared likely. Our results suggest that identical patterns in local-versus-regional species richness observed on two different spatial scales arise via different mechanisms: infracommunities appear saturated with flea species most likely because of local processes, such as host immune defenses, whereas component communities are saturated with species through interspecific competition, possibly among larval stages.

Animals↗

Sex-biased parasitism, seasonality and sexual size dimorphism in desert rodents.

We investigated seasonality of gender differences in the patterns of flea infestation in nine rodent species to test if sex-biased parasitism in terms of mean abundance, species richness, prevalence and the level of aggregation (a) varies among hosts and between seasons, and (b) is linked to sexual size dimorphism. Sexual size differences were significant in both summer and winter in Acomys cahirinus, Gerbillus pyramidum and Meriones crassus, and in winter only in Acomys russatus, Gerbillus dasyurus, Gerbillus nanus and Sekeetamys calurus. Sexual size dimorphism was male biased except for A. russatus in which it was female biased. Manifestation of sexual differences in flea infestation was different among hosts between seasons. A significant effect of sex on mean flea abundance was found in six hosts, on mean flea species richness in five hosts and on prevalence in two hosts. Male-biased parasitism was found in summer in one host only and in winter in five hosts. Female-biased parasitism occurred in winter in A. russatus. Gender differences in the slopes of the regressions of log-transformed variances against log-transformed mean abundances occurred in three hosts. No relationship was found between sexual size dimorphism and any parasitological parameter in any season using both conventional regressions and the method of independent contrasts. Our results suggest that sex-biased parasitism is a complicated phenomenon that involves several different mechanisms.

Animals↗

Evolution of host specificity in fleas: is it directional and irreversible?

Evolutionary trends in the evolution of host specificity have been the focus of much discussion but little rigorous empirical testing. On the one hand, specialization is often presumed to lead irreversibly into evolutionary dead ends and little diversification; this would mean that generalists might evolve into specialists, but not vice versa. On the other hand, low host specificity may limit the risk of extinction and provide more immediate fitness benefits to parasites, such that selection may favour evolution toward a generalist strategy. Here, we test for directionality in the evolution of host specificity using a large data set and phylogenetic information on 297 species of fleas parasitic on small mammals. The analyses determined whether host specificity, measured both as the number of host species exploited and their taxonomic diversity, was related to clade rank of the flea species, or the number of branching events between an extant species and the root of the phylogenetic tree (i.e., the total path length from the root of the tree to the species). Based on regression analyses, we found positive relationships between the number of host species used and clade rank across all 297 species, as well as within one (Hystrichopsyllidae) of four large families and one of seven large genera investigated separately; in addition, we found a positive relationship between the taxonomic diversity of host species used and clade rank in another of the seven genera. These results suggest a slight evolutionary trend of decreasing host specificity. Using a much more conservative likelihood ratio test, however, a random walk, or null model, of evolution could not be discarded in favour of the directional trends in all cases mentioned above. Still, these results suggest that host specificity may have tended to decrease in many flea lineages, a process that could have been driven by the benefits of exploiting a wide range of host species.

Adaptation, Biological↗

What are the factors determining the probability of discovering a flea species (Siphonaptera)?

Our aim was to determine which of four variables (number of host species exploited by the parasite, taxonomic distinctness of these hosts, geographic range of the principal host, and year of description of this host) was the best predictor of description date of fleas. The study used previously published data on 297 flea species parasitic on 197 species of small mammals from 34 different regions of the Holarctic and one region from the Neotropics. We used both simple linear and multiple regressions to evaluate the relationships between the four predictor variables and the year of flea description, on species values as well as on phylogenetically independent contrasts. Whether or not the analyses controlled for flea phylogeny, all predictor variables correlated significantly with year of flea description when tested separately. In multiple regressions, however, the number of exploited host species was the best predictor of the date of flea description, with the geographic range of the principal host species as well as the date of its description having a lesser, though significant, influence. Overall, our results indicate that a flea species is more likely to be discovered and described early if its biological characteristics (exploitation of many host species) and those of its hosts (long-known to science, broad geographic distributions) increase its chances of being included in a collection. Because the variables we investigated only explained about 10-11% of the variation in year of description among flea species, other factors must be important, such as temporal variability in the activity of flea taxonomists.

Animals↗

Distribution of fleas (Siphonaptera) among small mammals: mean abundance predicts prevalence via simple epidemiological model.

We used data on the abundance and distribution of fleas parasitic on small mammals in Slovakia and aimed: (i) to confirm a positive relationship between abundance and distribution fleas within and across host species; and (ii) to test if prevalence of fleas can be reliably predicted from a simple epidemiological model that takes into account flea mean abundance and its variance. Prevalence of a flea species increased with an increase in its mean abundance both within and across host species. We calculated prevalences both for each flea-host association and for each flea species across all hosts. Observed prevalences did not differ significantly from those predicted by the epidemiological model using parameters of Taylor's power relationship between mean abundance of fleas and its variance. Regressions of predicted prevalences against observed prevalences produced slope values that did not differ significantly from unity and were independent of scale (within or across host species). Our results demonstrated that up to 96% of variance in flea prevalence can be explained solely by their mean abundance. We concluded that, in general, there is no need to invoke other, more complex factors for the explanation of the variation in flea prevalence.

Animals↗

Energy costs of blood digestion in a host-specific haematophagous parasite.

Fleas consume and digest blood from their hosts. We hypothesized that the energy costs of digestion of blood by fleas is dependent on the host species. To test this hypothesis, we studied CO2 emission, a measure of energy expenditure, during digestion of a blood meal taken by Parapulex chephrenis from a preferred (Acomys cahirinus) and a non-preferred (Gerbillus dasyurus) host. We predicted that the energy cost of digestion would be lower for A. cahirinus blood than that for G. dasyurus. Male and female fleas consumed similar amounts of blood per unit body mass, independent of host species. Our prediction was supported in that fleas expended significantly more energy digesting blood of G. dasyurus than blood of A. cahirinus. We also found CO2 emission rates of fed fleas were higher than those of unfed fleas and differed significantly among stages of blood digestion when a flea fed on G. dasyurus but not when it fed on A. cahirinus. When fed on G. dasyurus, fleas spent less energy during earlier than later stages of digestion.

Analysis of Variance↗

Ectoparasitic "jacks-of-all-trades": relationship between abundance and host specificity in fleas (Siphonaptera) parasitic on small mammals.

Animal species with larger local populations tend to be widespread across many localities, whereas species with smaller local populations occur in fewer localities. This pattern is well documented for free-living species and can be explained by the resource breadth hypothesis: the attributes that enable a species to exploit a diversity of resources allow it to attain a broad distribution and high local density. In contrast, for parasitic organisms, the trade-off hypothesis predicts that parasites exploiting many host species will achieve lower mean abundance on those hosts than more host-specific parasites because of the costs of adaptations against multiple defense systems. We test these alternative hypotheses with data on host specificity and abundance of fleas parasitic on small mammals from 20 different regions. Our analyses controlled for phylogenetic influences, differences in host body surface area, and sampling effort. In most regions, we found significant positive relationships between flea abundance and either the number of host species they exploited or the average taxonomic distance among those host species. This was true whether we used mean flea abundance or the maximum abundance they achieved on their optimal host. Although fleas tended to exploit more host species in regions with either larger number of available hosts or more taxonomically diverse host faunas, differences in host faunas between regions had no clear effect on the abundance-host specificity relationship. Overall, the results support the resource breadth hypothesis: fleas exploiting many host species or taxonomically unrelated hosts achieve higher abundance than specialist fleas. We conclude that generalist parasites achieve higher abundance because of a combination of resource availability and stability.

Animals↗

Immune responses to fleas in two rodent species differing in natural prevalence of infestation and diversity of flea assemblages.

We studied in vivo immune response to fleas in two gerbils, Gerbillus dasyurus and Gerbillus andersoni allenbyi, which differed in their natural species richness of flea assemblages and prevalence of infestation. G.dasyurus is parasitized naturally by several flea species, but the prevalence of infestation is low, whereas G. a. allenbyi is parasitized by a single flea species, with high prevalence of infestation. We hypothesized that immunological parameters and the cell-mediated specific immune response to an antigen from an unfamiliar flea species differ between the two gerbil species. Parasitized and control gerbils of both species demonstrated similar, relatively low levels of spontaneous glucose consumption. The same was true for the phytohemagglutinin treatment. Responses to antigen from unfamiliar flea species were higher than both spontaneous glucose consumption and response to phytohemagglutinin in parasitized and control G. a. allenbyi and parasitized G. dasyurus. However, no significant difference in the spontaneous blast transformation index and responses to both phytohemagglutinin and flea antigen was found in control G. dasyurus. The number of white blood cells was significantly lower in control than in parasitized G. dasyurus, whereas no difference in the number of white blood cells was found between control and parasitized G. a. allenbyi. The levels of circulating immune complexes and concentrations of immunoglobulins did not differ between parasitized and control individuals in both species. Phagocytic activity was significantly higher in males than in females of G. a. allenbyi but not of G. dasyurus. In addition, phagocytes of G. dasyurus appeared to be significantly more active than those of G. a. allenbyi.

Animals↗

Immune response to fleas in a wild desert rodent: effect of parasite species, parasite burden, sex of host and host parasitological experience.

We studied immune responses of the jird Meriones crassus to different flea species belonging to the same family. We used jirds maintained in an outdoor enclosure (enclosure; N=18) and parasitized by fleas Xenopsylla conformis mycerini and Xenopsylla ramesis, and also jirds born in the laboratory to previously parasitized mothers (laboratory animals; N=23). We asked (i) whether cross-immunity to different fleas occurs, (ii) whether there is a sex difference in immune responses to flea parasitism and (iii) whether the severity of the immune responses depends on parasite load. In the enclosure animals, immune response to antigen from the unfamiliar flea Synosternus cleopatrae pyramidis did not differ from those to antigens from the familiar fleas. In contrast, laboratory rodents demonstrated no difference in the immune response between S. c. pyramidis antigen and either the phytohemagglutinin treatment or controls, although their responses to antigens of fleas familiar to their mothers (X. c. mycerini and X. ramesis) were significantly higher than those to antigen of S. c. pyramidis and phytohemagglutinin. The results clearly demonstrated that (i) cross-reactivity in rodent responses to different flea species occurred for enclosure but not for laboratory jirds and (ii) immune-naïve animals whose mothers were parasitized by fleas had some degree of immunity against fleas. The only sex difference in immunological parameters was the higher level of circulating immune complexes in females than in males. Only phagocytic activity was affected by flea burden, decreasing with an increase in flea numbers.

Analysis of Variance↗