The influence of the host on the frequency of block formation in males and females of Citellophilus tesquorum altaicus Ioff, 1936 (Siphonaptera: Ceratophyllidae) and transmission of the plague pathogen by these fleas

Data obtained during feeding of infested females and males of Citellophilus tesquorum altaicus Ioff, 1936 (Siphonaptera: Ceratophyllidae), the main vector of plague in Tuva natural plague focus, on the natural hosts and on laboratory animals were analyzed. Sexual differences in fleas were found to depend on the host. Females fed more actively on longtailed ground squirrels Spermophilus undulatus than on white mice. The feeding activity of males on the two hosts did not differ. A higher mortality of infested females and males was observed during feeding on mice. The frequency of block formation and pathogen transmission in males was higher during bloodsucking on the ground squirrel; in females, during feeding on mice. Thus, sex-related differences in the plague pathogen transmission revealed in laboratory white mice may be not the same in nature. This should be taken into account when extrapolating the experimental data on the natural relations between the plague pathogen and ectoparasites.     

The Effect of Initial Infection of <Emphasis Type="Italic">Citellophilus tesquorum altaicus</Emphasis> Ioff, 1936 (Siphonaptera: Ceratophyllidae) with <Emphasis Type="Italic">Yersinia pestis</Emphasis> on Its Alimentary Activity,Mortality, and Biofilm Formation

Alimentary activity and mortality was assessed in fleas Citellophilus tesquorum altaicus non-infected with Yersinia pestis and those with initial infection levels 50 and 100% during feeding on a non-specific host (white mice). The presence of the plague pathogen in fleas significantly stimulated their feeding activity, especially in females. No effect of infection on flea mortality was observed. At the same time, male fleas died more frequently than females.     

Factors of Yersinia pestis providing circulation and persistence of plague pathogen in ecosystems of natural foci. Communication 2

To maintain continuous circulation of plague pathogen in natural foci, the pathogen should be capable of invading host organism, resisting the bactericide protective systems of rodent, and reproducing itself to maintain the content of bacteria at a level sufficient for further transmission by fleas to a new host. Each of these stages of the Yersinia pestis circulation is determined by a variety of factors of plague pathogen, which may act either individually or in combination. Each of the factors itself may be involved in the pathological process at different stages of its development or in pathogen transmission. However, it is only the aggregate of the factors (regardless of significant or insignificant individual contribution to the sum effect) that provides persistence of plague pathogen in natural foci. The plague pathogen factors providing its transmission from one host organism to the next as well as correlation of individual factors of pathogensis and expression of various household genes with plague pathogensis virulence are considered in the second communication. This review was compiled on the basis of not only well-known works but also some sources of limited availability, particularly, for English-speaking audience.     

Polymerase chain reaction (PCR) identification of rodent blood meals confirms host sharing by flea vectors of plague

Elucidating feeding relationships between hosts and parasites remains a significant challenge in studies of the ecology of infectious diseases, especially those involving small or cryptic vectors. Black‐tailed prairie dogs (Cynomys ludovicianus) are a species of conservation importance in the North American Great Plains whose populations are extirpated by plague, a flea‐vectored, bacterial disease. Using polymerase chain reaction (PCR) assays, we determined that fleas (Oropsylla hirsuta) associated with prairie dogs feed upon northern grasshopper mice (Onychomys leucogaster), a rodent that has been implicated in the transmission and maintenance of plague in prairie‐dog colonies. Our results definitively show that grasshopper mice not only share fleas with prairie dogs during plague epizootics, but also provide them with blood meals, offering a mechanism by which the pathogen, Yersinia pestis, may be transmitted between host species and maintained between epizootics. The lack of identifiable host DNA in a significant fraction of engorged Oropsylla hirsuta collected from animals (47%) and prairie‐dog burrows (100%) suggests a rapid rate of digestion and feeding that may facilitate disease transmission during epizootics but also complicate efforts to detect feeding on alternative hosts. Combined with other analytical approaches, e.g., stable isotope analysis, molecular genetic techniques can provide novel insights into host‐parasite feeding relationships and improve our understanding of the role of alternative hosts in the transmission and maintenance of disease.     

Blood-feeding of the gerbil flea Nosopsyllus laeviceps kuzenkovi (Yagubyants), vector of plague in Inner Mongolia,China

Abstract. The gerbil flea Nosopsyllus laeviceps (Wagner) (Siphonaptera: Ceratophyllidae) is an important Palaearctic vector of enzootic plague among sylvan and campestral rodents. Laboratory blood-feeding experiments with the subspecies N.l.kuzenkovi (Yagubynts) from Inner Mongolia, China, using three rodent species including the natural host Meriones unguiculatus (Milne-Edwards), demonstrated a positive relationship between per cent feeding and time allowed for feeding, particularly for female fleas. Also the bloodmeal size was proportional to sucking time and was significantly greater in females, which are larger, than in males. However, on all four strains of host tested, there was no significant correlation between the feeding success rate of the fleas and the amount of blood they imbibed. Fleas fed better on active compared to confined M.unguiculatus. These findings agree with studies of other flea species.     

The Effect of Male Ticks on the Feeding Performance of Immature Stages of Rhipicephalus Sanguineus and Amblyomma Americanum (Acari: Ixodidae)

A unique group of immunoglobulin-binding proteins (IGBPs), produced by ixodid male ticks during the latter half of their prolonged feeding period, improves the feeding performance of co-feeding females. As a follow-up to this observation, we investigated whether male tick feeding also affects the feeding of other developmental stages. Immature stages of Rhipicephalus sanguineus (Latreeille) and Amblyomma americanum (L.) were fed on rabbits in the presence or absence of conspecific males. The mean weight of larvae and nymphs of both species that fed around males and detached from the host on the first day of dropping was significantly higher than when the immature ticks fed on rabbits in the absence of males. However, larvae of both species and nymphs of R. sanguineus that fed slower and detached on the second day of dropping did not show significant differences in weight. A similar pattern was observed for A. americanum nymphs although, unlike R. sanguineus, the presence of males also influenced the feeding performance of the nymphs that fed slowly and detached on the second day of drop-off. The improved feeding performance demonstrated by immature ticks in the presence of males may be due to immunomodulatory saliva proteins, such as immunoglobulin-binding proteins (IGBPs) that are introduced into the co-feeding site. The results are considered in relation to the distribution of ixodid tick species on their natural hosts.     

Interactions between host sex and age of exposure modify the virulence–transmission trade‐off

The patterns of immunity conferred by host sex or age represent two sources of host heterogeneity that can potentially shape the evolutionary trajectory of disease. With each host sex or age encountered, a pathogen's optimal exploitative strategy may change, leading to considerable variation in expression of pathogen transmission and virulence. To date, these host characteristics have been studied in the context of host fitness alone, overlooking the effects of host sex and age on the fundamental virulence–transmission trade‐off faced by pathogens. Here, we explicitly address the interaction of these characteristics and find that host sex and age at exposure to a pathogen affect age‐specific patterns of mortality and the balance between pathogen transmission and virulence. When infecting age‐structured male and female Daphnia magna with different genotypes of Pasteuria ramosa, we found that infection increased mortality rates across all age classes for females, whereas mortality only increased in the earliest age class for males. Female hosts allowed a variety of trade‐offs between transmission and virulence to arise with each age and pathogen genotype. In contrast, this variation was dampened in males, with pathogens exhibiting declines in both virulence and transmission with increasing host age. Our results suggest that differences in exploitation potential of males and females to a pathogen can interact with host age to allow different virulence strategies to coexist, and illustrate the potential for these widespread sources of host heterogeneity to direct the evolution of disease in natural populations.     

Visualizing Non Infectious and Infectious Anopheles gambiae Blood Feedings in Naive and Saliva-Immunized Mice

Background

Anopheles gambiae is a major vector of malaria and lymphatic filariasis. The arthropod-host interactions occurring at the skin interface are complex and dynamic. We used a global approach to describe the interaction between the mosquito (infected or uninfected) and the skin of mammals during blood feeding.

Methods

Intravital video microscopy was used to characterize several features during blood feeding. The deposition and movement of Plasmodium berghei sporozoites in the dermis were also observed. We also used histological techniques to analyze the impact of infected and uninfected feedings on the skin cell response in naive mice.

Results

The mouthparts were highly mobile within the skin during the probing phase. Probing time increased with mosquito age, with possible effects on pathogen transmission. Repletion was achieved by capillary feeding. The presence of sporozoites in the salivary glands modified the behavior of the mosquitoes, with infected females tending to probe more than uninfected females (86% versus 44%). A white area around the tip of the proboscis was observed when the mosquitoes fed on blood from the vessels of mice immunized with saliva. Mosquito feedings elicited an acute inflammatory response in naive mice that peaked three hours after the bite. Polynuclear and mast cells were associated with saliva deposits. We describe the first visualization of saliva in the skin by immunohistochemistry (IHC) with antibodies directed against saliva. Both saliva deposits and sporozoites were detected in the skin for up to 18 h after the bite.

Conclusion

This study, in which we visualized the probing and engorgement phases of Anopheles gambiae blood meals, provides precise information about the behavior of the insect as a function of its infection status and the presence or absence of anti-saliva antibodies. It also provides insight into the possible consequences of the inflammatory reaction for blood feeding and pathogen transmission.     

The Deer Mouse (<Emphasis Type="Italic">Peromyscus maniculatus</Emphasis>) as an Enzootic Reservoir of Plague in California

It has long been theorized that deer mice (Peromyscus maniculatus) are a primary reservoir of Yersinia pestis in California. However, recent research from other parts of the western USA has implicated deer mice as spillover hosts during epizootic plague transmission. This retrospective study analyzed deer mouse data collected for plague surveillance by public health agencies in California from 1971 to 2016 to help elucidate the role of deer mice in plague transmission. The fleas most commonly found on deer mice were poor vectors of Y. pestis and occurred in insufficient numbers to maintain transmission of the pathogen, while fleas whose natural hosts are deer mice were rarely observed and even more rarely found infected with Y. pestis on other rodent hosts. Seroprevalence of Y. pestis antibodies in deer mice was significantly lower than that of several chipmunk and squirrel species. These analyses suggest that it is unlikely that deer mice play an important role in maintaining plague transmission in California. While they may not be primary reservoirs, results supported the premise that deer mice are occasionally exposed to and infected by Y. pestis and instead may be spillover hosts.     

Hyperparasitism in Ornithodoros erraticus

Hyperparasitism is common in laboratory colonies of Egyptian Ornithodoros erraticus whether the ticks are on or off the rodent host. Sex, recent engorgement, and size appear to be major factors in this feeding relationship. Males, and nymphs that produced males (N-male), parasitized females and nymphs that produced females (N-female) more frequently than they parasitized males and N-male. Females and N-female seldom parasitized females and N-female and did not parasitize males or N-male. Engorging and recently engorged, large ticks attracted smaller unfed ones. No preferred attachment site was observed. Hyperparasitizing females and parasitized females weighed less, had a longer preoviposition period, and produced fewer eggs than normally feeding, unparasitized females. About 30% and 80% of the females parasitized by males and females, respectively, died within 3 mo; only 7% of unparasitized females died within the same period. The proximity of replete or engorging ticks may be sufficient stimulus for hyperparasitism. Additional factors may include production of an attracting kairomone by fed ticks or the absence under certain conditions of a deterrent to hyperparasitism normally produced by fed ticks. Borrelia crocidurae spirochetes are transmitted during hyperparasitism.     

Effects of a lipid-rich diet on adult parasitoid income resources and survival

Effectiveness of parasitoids as natural enemies in agro-ecosystems depends on key traits such as fecundity and longevity. Energy sources allocated into survival and reproduction can be mobilized from capital stores acquired during larval feeding, or from income resources through adult feeding. Adult parasitoids have a restricted diet consisting solely of carbohydrates and most species do not replenish lipid levels after emergence from their host. Here, we have adopted a novel approach that could improve pest elimination by parasitoids, in which we added different ratios of olive oil to the food of adults to reveal the potential of a lipid-rich diet to increase lipid levels and allocation into survival. Our results show that males of the parasitoid Cotesia glomerata had higher lipid levels when fed a diet containing a ratio of 90:10% honey to olive oil when compared to males at emergence and males fed honey-only. For females, lipid levels at emergence were similar to those of females fed a diet supplemented with a ratio of 75:25% honey to olive oil, yet in both of these treatments lipid levels were significantly higher when compared to females fed honey-only. This suggests that females on a lipid-containing diet economized on their lipid use. In contrast, the survival of males and females was negatively affected by the addition of olive oil and no differences in survival were found when wasps were fed 1 day on various ratios of honey to olive oil compared to honey-only. Our results show that the addition of a lipid source can increase or maintain nutrient availability and further research into lipid supplementation could minimize detrimental effects in order to increase the effectiveness of parasitoids in certain agro-ecosystems.     

母代高脂饮食诱导子代在小鼠生命早期出现糖脂代谢紊乱且具有雄性易感性

目的：探讨孕期和哺乳期的高脂饮食能否导致子代在生命早期出现糖脂代谢紊乱。方法成年雌性C57BL/6J小鼠与正常饮食雄性小鼠进行交配,孕鼠随机分为高脂饮食组和正常饮食组,在孕期和哺乳期喂养高脂饲料或正常饲料,至交配后第一代鼠断乳时（3周龄）观察其糖脂代谢相关性指标以及肝脏病理表现。结果较正常饮食组子鼠相比,高脂饮食子鼠出生体重更低（ P＜0.05）。在断乳时,高脂饮食组雄性子鼠体重较重（ P ＝0.038）,腹腔糖耐量实验30 min和60 min血糖明显升高（P值分别为＜0.001和＜0.01）,糖耐量曲线下面积较大（P＝0.0016）,HOMA-IR值较大（P＜0.05）,雌性子鼠腹腔糖耐量实验在30 min血糖高于正常组（P＜0.01）,而糖耐量曲线下面积和HOMA-IR值在两组之间无明显统计学意义。雄性和雌性子代小鼠空腹胆固醇水平高脂饮食组均高于正常饮食组（ P值分别为＜0.0001和0.0004）,而两组雄性和雌性子代小鼠空腹胰岛素和甘油三酯水平差异均无显著性（ P均＞0.05）。另外,在断乳时高脂饮食子鼠出现肝脏脂肪变性,雌性和雄性子鼠无明显差异。结论母鼠孕期和哺乳期高脂饮食能够诱导子代在生命早期就能出现糖脂代谢紊乱并且雄性子鼠更易出现肥胖、糖耐量异常、胰岛素抵抗。     

Ability of transovarially and subsequent transstadially infected Ixodes hexagonus ticks to maintain and transmit Borrelia burgdorferi in the laboratory

In a previous study, transstadial and transovarial survival of Borrelia burgdorferi in Ixodes hexagonus and transmission to laboratory mice via the bite of infected females were demonstrated. Here, we report the ability of I. hexagonus progeny infected transovarially to maintain and transmit the spriochaete to the host.Ticks were examined for spirochaetes by direct immunofluorescence antibody test. I. hexagonus larvae derived from the parental transstadially infected females were fed on two white mice: 21/54 (38.9%) of these ticks examined as unfed nymphs were infected. I. hexagonus nymphs were fed on three white mice and examined for spirochaetes after moulting as adults: 7/25 (28%) were found to harbour the spirochaete. The success of B. burgdorferi transmission to the mice by larval and nymphal I. hexagonus was determined by xenodiagnosis using I. ricinus larvae: 20/50 (40%) and 30/99 (30.3%) of the I. ricinus larvae fed on the mice infected by I. hexagonus larvae and nymphs respectively became infected.This study shows that B. burgdorferi can be maintained through transovarial and subsequent transstadial transmissions in I. hexagonus.     

Experimental transmission of Leishmania tropica to hamsters and mice by the bite of Phlebotomus sergenti

Phlebotomus sergenti is a natural vector of Leishmania tropica. However, the ability of P. sergenti to transmit L. tropica by bite has not been proven experimentally yet. We have transmitted L. tropica to golden hamsters and BALB/c mice by the bite of P. sergenti. Sand flies and Leishmania both originated from an anthroponotic cutaneous leishmaniasis focus in Urfa, Turkey. P. sergenti females from a laboratory colony were infected by feeding on lesions of needle-inoculated hamsters or mice. Gravid females were allowed to refeed on uninfected hosts 9-15 d after the infective feeding. At the second feeding, some infected females took a full blood meal, while others only a partial one; some females failed to feed at all. The ability of infected females to take a blood meal did not correlate with the parasite transmissibility. In four BALB/c mice, lesions developed after 1-6 months. In two albino hamsters (Mesocricetus auratus), lesions developed 1 month after the infective feeding, and Leishmania could be reisolated from these sites. Another hamster did not develop a lesion; however, the feeding site and the adjacent ear were PCR positive 1 year after infective feeding. Our results show that dissemination to other parts of host body occurs in L. tropica after sand fly bite. Experimental transmission of the parasite confirms that P. sergenti is a natural vector of L. tropica.     

Linking predator–prey interactions with exposure to a trophically transmitted parasite using PCR‐based analyses

Parasite transmission is determined by the rate of contact between a susceptible host and an infective stage and susceptibility to infection given an exposure event. Attempts to measure levels of variation in exposure in natural populations can be especially challenging. The level of exposure to a major class of parasites, trophically transmitted parasites, can be estimated by investigating the host's feeding behaviour. Since the parasites rely on the ingestion of infective intermediate hosts for transmission, the potential for exposure to infection is inherently linked to the definitive host's feeding ecology. Here, we combined epidemiological data and molecular analyses (polymerase chain reaction) of the diet of the definitive host, the white‐footed mouse (Peromyscus leucopus), to investigate temporal and individual heterogeneities in exposure to infection. Our results show that the consumption of cricket intermediate hosts accounted for much of the variation in infection; mice that had consumed crickets were four times more likely to become infected than animals that tested negative for cricket DNA. In particular, pregnant female hosts were three times more likely to consume crickets, which corresponded to a threefold increase in infection compared with nonpregnant females. Interestingly, males in breeding condition had a higher rate of infection even though breeding males were just as likely to test positive for cricket consumption as nonbreeding males. These results suggest that while heterogeneity in host diet served as a strong predictor of exposure risk, differential susceptibility to infection may also play a key role, particularly among male hosts. By combining PCR analyses with epidemiological data, we revealed temporal variation in exposure through prey consumption and identified potentially important individual heterogeneities in parasite transmission.     

The nutritional contribution of males affects the feeding behavior and spatial distribution of females in a bruchid beetle,<Emphasis Type="Italic"> Bruchidius dorsalis</Emphasis>

The relationship between the quality/quantity of male investment and the feeding behavior of females was investigated in a bruchid weevil, Bruchidius dorsalis (Fahraeus), whose males donate nutrition via seminal fluid to females. Experiments on the effect of feeding regimes of both sexes on the mating frequency of females showed that females mated at a higher frequency if given low-quality food or poor male investment. On the other hand, the experiment that examined the effect of male investment quality on female feeding behavior showed that females receiving the high-quality investment exhibited feeding behavior less often. These results suggest that male investment and feeding behavior play the same role for B. dorsalis females. These experiments also showed that there are sex-related asymmetries in mating and feeding behaviors: females mated more often but males fed more often. Moreover, a field census suggested that only males visited non-host flowers to feed on the pollen and nectar during the non-flowering period of the host plants; females always stayed on the host plants irrespective of the flowering phenology. These results suggest that in B. dorsalis courtship role reversal and sex-specific feeding modes are fairly fixed and obligatory, and that male investment, derived from sexual selection, could affect the feeding behavior and spatial distribution of both sexes, which may have far-reaching impact in various ecological contexts.     

Interstadial and Infestation Level-dependent Variation in the Transmission Efficiency of Borrelia Burgdorferi from Mice to Ixodes Ricinus Ticks

The efficiency with which the spirochaete Borrelia burgdorferi sensu stricto was transmitted from laboratory mice to larval and nymphal Ixodes ricinus ticks was assessed, using the polymerase chain reaction. The transmission efficiency to nymphs was significantly greater than to larvae when both fed together on the same host. Increased tick infestation levels of mice were correlated with significantly greater engorgement weights and higher B. burgdorferi transmission coefficients from mice to nymphs. These observations indicate that both the feeding success of ticks and the transmission coefficients from host to tick may be influenced by the tick infestation level of an infected host. The infestation level and the relative numbers of each life stage of the tick are factors which should be considered in the design of transmission experiments.     

Microarray analysis of gene expression changes in feeding female and male lone star ticks,Amblyomma americanum (L)

A collection of EST clones from female tick Amblyomma americanum salivary glands was hybridized to RNA from different feeding stages of female tick salivary glands and from unfed or feeding adult male ticks. In the female ticks, the expression patterns changed dramatically upon starting feeding, then changed again towards the end of feeding. On beginning feeding, genes possibly involved in survival on the host increased in expression as did many housekeeping genes. As feeding progressed, some of the survival genes were downregulated, while others were upregulated. When the tick went into the rapid feeding phase, many of the survival genes were downregulated, while a number of transport‐associated genes and genes possibly involved in organ degeneration increased. In the males, the presence of females during feeding made a small difference, but feeding made a larger difference. Males showed clear differences from females in expression, as well. Protein synthesis genes were expressed more in all male groups than in the partially fed females, while the putative secreted genes involved in avoiding host defenses were expressed less. © 2009 Wiley Periodicals, Inc.     

Attraction to Host Plant Volatiles and Feeding Performance of <Emphasis Type="Italic">Naupactus Xanthographus</Emphasis> (Coleoptera: Curculionidae) is Affected by Starvation

The grape weevil, Naupactus xanthographus Germar (Coleoptera: Curculionidae), is a polyphagous insect which is a cause of important damage to several economically relevant crops, including grape (Vitis vinifera) and avocado (Persea americana), in several countries of Latin America. The larvae cause damage to the roots and rootlets of plants, and adults feed on leaves of their host plant. Despite its economic importance, there are few reports on the behavioral and nutritional ecology of this weevil. In this context, laboratory feeding and olfactometer bioassays with N. xanthographus were performed. The feeding performance was evaluated by measuring the weight variation of the insects after 1 and 6 h of feeding on grape or avocado leaves, respectively. After 1 h of feeding, insects showed no significant differences in weight increase. However, after a period of 6 h of feeding, males had continued feeding on grape leaves, but not on avocado leaves. Bioassays using a Y-tube olfactometer showed that males are attracted to volatiles of both host plants. Furthermore, starved males and females showed no preference to volatiles of grape or avocado. However, non-starved males and females preferred grape volatiles over avocado volatiles. Based on the combined results of the assays, we conclude that grape is preferred over avocado for N. xanthographus. Furthermore, this is the first report on the effect of starvation on the attraction to host plant volatiles in Curculionidae.     

Development and field application of a molecular probe for the primary pathogen of the coral disease white plague type II

One of the current problems in the field of coral disease research is that of tracking coral pathogens in the natural environment. A promising method to do this is by use of pathogen-specific molecular probes. However, this approach has been little used to date. We constructed, and validated in the laboratory, a fluorochrome-labeled molecular probe specific to Aurantimonas coralicida, the bacterial pathogen of the Caribbean coral disease white plague type II (WPIl). We then used the probe to test field samples of diseased coral tissue for the presence of this pathogen. Probe design was based on a unique subset (25 nucleotides) of the complete l6S rRNA gene sequence derived from a pure culture of the pathogen. The pathogen-specific probe was labeled with the fluorochrome GreenStar* FITC (fluorescein isothiocyanate, GeneDetect Ltd, New Zealand). As a control, we used the universal eubacterial probe EUB 338, labeled with a different fluorochrome (TRITC, tetra-methylrhodamine isothiocyanate). Both probes were applied to laboratory samples of pure cultures of bacteria, and field samples collected from the surface of the disease line of corals exhibiting signs of white plague (types I and II), healthy controls, and corals with an uncharacterized disease ("patchy necrosis"). All samples were analyzed using fluorescence in situ hybridization (FISH). We have determined that the probe is specific to our laboratory culture of the coral pathogen, and does not react with other bacterial species (the eubacterial probe does). The WPII pathogen was detected in association with diseased coral samples collected from coral colonies on reefs of the Bahamas (n= 9 samples) exhibiting signs of both WPI and WPII. Diseased (and healthy) tissue samples (n- 4) from corals exhibiting signs of "patchy necrosis" were also assayed. In this case the results were negative, indicating that the same pathogen is not involved in the two diseases. Incorporation and use of pathogen-specific probes can significantly expand our knowledge of the etiology of coral diseases.