PubMed Health⌕ Search

Biomedical subjects

B H Noden

Publications and source records attributed to B H Noden.

16 recordsLinked to original sources

Molecular identification of Rickettsia typhi and R. felis in co-infected Ctenocephalides felis (Siphonaptera: Pulicidae).

Rickettsia typhi and R. felis, 2 closely related rickettsial species, often have been identified in cat fleas, Ctenocephalides felis (Bouché) from the same geographical location. However, no fleas have been found to be naturally infected concurrently with both rickettsial species. To examine whether one rickettsial species can develop simultaneously with another species in the same flea host, cat fleas, naturally infected with R. felis, were allowed to feed on blood containing R. typhi (Ethiopian strain, 10(7) PFU/ml). Experimental controls consisted of uninfected cat fleas, fleas infected with only R. typhi; and fleas naturally infected with only R. felis. After 9 d at 28 degrees C, the fleas were examined by PCR amplification and subsequent restriction digest analysis and dot blot hybridization of PCR products. Results from these studies demonstrated that R. felis and R. typhi are capable of co-existing in the same flea host. Subsequent isolation of R. typhi from dually infected fleas by tissue culture indicated that R. typhi was viable and capable of being maintained in fleas naturally infected with R. felis. As more studies confirm the presence of R. felis and R. typhi in domestic pets and peridomestic vertebrates in urban areas, the ability of the individual cat fleas, which live on these animals, to support both rickettsial species could be an epidemiologically important consideration.

Animals↗

The impact of variations in temperature on early Plasmodium falciparum development in Anopheles stephensi.

The effect of temperature on early Plasmodium falciparum development was examined in Anopheles stephensi. The rates of both ookinete development and bloodmeal digestion were lengthened as temperatures decreased from 27 to 21 degrees C. However, low temperatures (21-27 degrees C) did not significantly influence infection rates or densities of either ookinetes or oocytes. In contrast, high temperatures (30 and 32 degrees C) significantly impacted parasite densities and infection rates by interfering with developmental processes occurring between parasite fertilization and ookinete formation, especially during zygote and early ookinete maturation. This study demonstrates clearly that temperature affects the sporogonic development of P. falciparum in anophelines by altering the kinetics of ookinete maturation. These studies not only confirm the ookinete as the key development stage affecting the probability of vector infectivity, they provide new insights into the epidemiology of P. falciparum infections.

Animals↗

An immunological factor that affects Anopheles gambiae survival.

High titers of antibodies against Anopheles gambiae midguts were produced in New Zealand rabbits to identify midgut targets for an antimosquito vaccine. The serum from one of 8 rabbits (designated R2B6) killed 71.6% (Abbott's adjusted % mortality) of An. gambiae within 7 days. Mosquitoes ingesting R2B6 serum were unable to absorb their blood meal nutrients, resulting in reduced oviposition and egg hatching rates. Anopheles stephensi and Anopheles arabiensis were also killed when ingesting R2B6 serum but Anopheles freeborni, Anopheles albimanus, and Aedes aegypti were not affected. The mosquitocidal factor was a relatively large molecule (> 100,000 MW) maintained at threshold levels in the sera and killing was complement independent. Mortality, however, was not IgG mediated, as determined by protein A-sepharose fractionation. This surprising finding confounds possibilities of using antibodies against whole mosquito midguts as a step in the development of antimosquito vaccines.

Aedes↗

Noninfectious sporozoites in the salivary glands of a minimally susceptible anopheline mosquito.

In studies to evaluate vector-malaria parasite relationships, we have found that Anopheles albimanus is minimally susceptible to the rodent malaria parasite Plasmodium yoelii. Normally, less than 10% of A. albimanus develop oocyst infections compared to 80-100% for Anopheles stephensi and Anopheles freeborni mosquitoes. Although sporozoites produced in A. albimanus invade the salivary glands, they are not infectious to BALB/c or ICR mice. In 11 experiments with sporozoites from A. albimanus, intravenous inoculations of up to 24,000 sporozoites in individual mice failed to produce host infections. In contrast, inoculation of 300 sporozoites obtained from the salivary glands of A. stephensi and A. freeborni always infected mice. The noninfectious sporozoites from A. albimanus were morphologically similar to the infectious sporozoites from A. stephensi and yielded 4+ circumsporozoite precipitin reactions when incubated with a monoclonal antibody against the circumsporozoite protein of P. yoelii. The presence of noninfectious sporozoites in the salivary glands of A. albimanus suggests that this minimally susceptible vector either possesses a toxic factor that abolishes sporozoite infectiousness or lacks a critical substance needed by the sporozoite to become infectious. Sporozoite infectiousness was neither attenuated by incubation of infectious sporozoites with A. albimanus salivary glands nor restored when noninfectious sporozoites were incubated with A. stephensi salivary glands. These studies provide a starting point for defining the biological basis of sporozoite infectivity.

Animals↗

Plasmodium falciparum: the population structure of mature gametocyte cultures has little effect on their innate fertility.

In vitro cultured Plasmodium falciparum gametocytes were fed to Anopheles gambiae (G3) mosquitoes to identify parasite population characteristics useful for predicting successful mosquito infections. Parameters were collected from an initial study of 90 infections over a two year period and a second study of 55 infections over 12 weeks. Parasite isolate/clone was identified as the most reliable predictor of gametocyte infectiousness. Parameters such as gametocyte age structure (stage IV:V ratio), exflagellation rate and macrogametocyte maturity were not reliable for predicting infectiousness but were useful for monitoring overall culture maturity. Other variables such as gametocyte density, chronological age of the culture at the time of feed, gametocyte sex ratio, asexual parasitemia, and mixing cultures before mosquito feeding were not predictive. Thus, if a reliable parasite isolate or clone is used, there is no need to measure other characteristics of in vitro gametocyte populations because these will not significantly improve one's ability to predict oocyst infection rates.

Animals↗

Prior blood feeding effects on susceptibility of Anopheles gambiae (Diptera: Culicidae) to infection with cultured Plasmodium falciparum (Haemosporida: Plasmodiidae).

We examined the relative susceptibilities of Anopheles gambiae Giles of different physiological ages to infection with cultured Plasmodium falciparum (Welch). Cohorts of mosquitoes were divided into three groups; one was fed uninfected blood on day 3 after emergence (i.e., one prior blood meal); another on days 3 and 7 after emergence (i.e., two prior blood meals); and a control group was maintained on sucrose. On days 10 to 12 after emergence, mosquitoes were fed human blood containing P. falciparum gametocytes. Prior blood feeding accelerated digestion of the infective blood meals and subtly altered susceptibility to infection with P. falciparum. When gametocyte cultures were highly fertile, all experimental groups were equally susceptible to infection. However, when gametocyte fertility was low, accelerated digestion had a detrimental effect on the transition of ookinetes to oocysts. Accelerated digestion may raise the threshold density of ookinetes required for the successful conversion of ookinetes to oocysts.

Animals↗

Exflagellation responses of cultured Plasmodium falciparum (Haemosporida: Plasmodiidae) gametocytes to human sera and midguts of anopheline mosquitoes (Diptera: Culicidae).

The process of exflagellation was quantified for cultured Plasmodium falciparum gametocytes exposed to human sera and midgut homogenates from six vector species of Anopheles mosquitoes. Neither serum factors related to malaria exposure nor factors in the midguts of taxonomically diverse anophelines had significant inhibitory effects on the exflagellation of P. falciparum microgametocytes. Therefore, differences in vector competence among anopheline species most likely are caused by vector-parasite interactions occurring after microgametogenesis.

Adult↗

Sporogonic development of cultured Plasmodium falciparum in six species of laboratory-reared Anopheles mosquitoes.

Sporogonic development of cultured Plasmodium falciparum was compared in six species of Anopheles mosquitoes. A reference species, A. gambiae, was selected as the standard for comparison. Estimates of absolute densities were determined for each lifestage. From these data, four aspects of parasite population dynamics were analyzed quantitatively: 1) successive losses in abundance as parasites developed from gametocyte to ookinete to oocyst stages, 2) oocyst production of sporozoites, 3) correlation between various lifestage parameters, and 4) parasite distribution. Parasite populations in A. gambiae incurred a 316-fold loss in abundance during the transition from macrogametocyte to ookinete stage, a 100-fold loss from ookinete to oocyst stage, yielding a total loss of approximately 31,600-fold (i.e., losses are multiplicative). Comparative susceptibilities in order were A. freeborni >> A. gambiae, A. arabiensis, A. dirus > A. stephensi, A. albimanus. The key transition(s) determining overall susceptibility differed among species. Despite species differences in oocyst densities and infection rates, salivary gland sporozoite production per oocyst (approximately 640) was the same among species. The most consistent association among lifestage parameters was a positive correlation between densities and infection rates of homologous lifestages. A curvilinear relationship between ookinete and oocyst densities in A. gambiae indicated a threshold density was required for ookinete conversion to oocysts (approximately 30 ookinetes per mosquito). The same relationship in A. freeborni was linear, with no distinct threshold. Ookinete and oocyst populations were negative binomially distributed in all species. Indices of heterogeneity in mosquito susceptibility to infection indicated that gene frequencies determining susceptibility fluctuated with time in all species, except A. freeborni where susceptibility remained homogenous throughout the study. This approach provides a framework for identifying mechanisms of susceptibility and evaluating Plasmodium sporogonic development in naturally occurring vector species in nature.

Analysis of Variance↗

Plasmodium falciparum: release of circumsporozoite protein by sporozoites in the mosquito vector.

The release of circumsporozoite (CS) protein by Plasmodium falciparum sporozoites was investigated to identify factors regulating this process within infected Anopheles gambiae mosquitoes. The potential for sporozoites to release CS protein in vitro was not dependent upon their site-specific developmental stage (i.e., mature oocysts, hemolymph, salivary glands), their duration in the vector, or their exposure to mosquito-derived components such as salivary glands or hemolymph. The capacity of sporozoites to release CS protein was depressed by mosquito blood feeding during periods of sporozoite migration to the salivary glands, but the effect was only temporary and those sporozoites already in the glands were not affected. Free CS protein in the salivary glands was present in 93.3% of 45 infective mosquitoes. Sporozoites from these same, individual mosquitoes were also tested in vitro for CS protein release. In both cases, the amount of soluble CS protein increased as a function of sporozoite density but the total amount of CS protein per sporozoite became progressively less with increasing numbers of sporozoites. Further experiments showed that sporozoite contact with increasing amounts of soluble CS protein caused a down-regulation of CS protein release. Thus, a primary factor regulating the production and release of CS protein by sporozoites is their contact with soluble CS protein within the mosquito.

Animals↗

Ingestion of Plasmodium falciparum sporozoites during transmission by anopheline mosquitoes.

We investigated the process of sporozoite transmission during blood feeding for Anopheles gambiae and An. stephensi experimentally infected with Plasmodium falciparum. When infective mosquitoes were fed 22-25 days postinfection on an anesthetized rat, sporozoites were detected in the midgut of 96.5% of 57 An. gambiae (geometric mean [GM] = 32.5, range 3-374) and in 96.2% of 26 An. stephensi (GM = 19.5, range 1-345). There were no significant differences between species either in salivary gland sporozoite loads or in the number of ingested sporozoites. There was a significant linear relationship between sporozoite loads and the numbers of ingested sporozoites for both An. gambiae (r = 0.38) and An. stephensi (r = 0.69). Subsequently, An. gambiae were tested for sporozoite transmission by allowing them to feed individually on a suspended capillary tube containing 10 microliters of blood. A total of 83.3% of 18 infective mosquitoes transmitted a GM of 5.9 (range 1-36) sporozoites. The same mosquitoes contained a GM of 23.4 (range 2-165) ingested sporozoites. The number of ingested sporozoites was related to sporozoite loads (r = 0.42) but not to the number of sporozoites ejected into capillary tubes. Ingested sporozoites remained in the midgut up to 10 hr after feeding. The comparable numbers of sporozoites ingested by infective mosquitoes in both experiments indicates that the actual number of sporozoites transmitted to the vertebrate host during blood feeding is significantly reduced by the blood ingestion process.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

Sporozoite transmission by Anopheles freeborni and Anopheles gambiae experimentally infected with Plasmodium falciparum.

A micro-membrane feeding technique was used to evaluate sporozoite transmission for Anopheles freeborni and An. gambiae experimentally infected with Plasmodium falciparum. From cohorts of infected mosquitoes with equivalent sporozoite loads, 75.9% of 29 An. freeborni transmitted a geometric mean (GM) of 4.9 sporozoites and 80% of 30 An. gambiae transmitted a GM of 11.3 sporozoites. Ingested sporozoites, in the blood meal immediately after feeding, were detected in 86.2% of 29 An. freeborni (GM = 9.0) and in 70% of 30 An. gambiae (GM = 44.1). Overall, sporozoites were transmitted and/or ingested by 90% of both species. Most infective mosquitoes transmitted < 1% of the total sporozoites in the salivary glands, and only up to 30% of the variation in transmission, ingestion, or total sporozoite output was related to sporozoite loads. The demonstration that An. gambiae transmitted more than twice as many sporozoites as An. freeborni is the first indication that vector species of anopheline mosquitoes differ in their innate potential for sporozoite transmission.

Animals↗

Population dynamics of Plasmodium falciparum sporogony in laboratory-infected Anopheles gambiae.

The population dynamics of cultured Plasmodium falciparum parasites was examined during their sporogonic development in Anopheles gambiae mosquitoes. Estimates of absolute densities were determined for each life stage, and life tables were constructed for each of 38 experimental infections. Macrogametocyte and ookinete mortalities contributed equally to the overall mortality. On average, there was a 40-fold decrease in parasite numbers in the transition from the macrogametocyte to the ookinete stage, a 69-fold decrease in the transition from ookinete to oocyst stages, and a total net decrease in parasite numbers from macrogametocyte to oocyst stage of 2,754-fold (i.e., multiplicative). There was no relationship between macrogametocyte and ookinete densities due to the inherent variability in fertility among different gametocyte cultures. There was a curvilinear relationship (r2 = 0.66) between ookinete and oocyst densities. Above a threshold of about 30 ookinetes/mosquito, the oocyst yield per ookinete became increasingly greater with increasing ookinete density. There was a linear relationship (r2 = 0.73) between oocyst and sporozoite densities, with an average of 663 salivary gland sporozoites produced per oocyst. Sporozoite production per oocyst was not affected by oocyst density and virtually all oocyst infections resulted in sporozoite infections of the salivery glands. This quantitative study indicates that the sporogony of cultured P. falciparum in laboratory-infected A. gambiae is an inefficient process and that the ookinete is the key transitional stage affecting the probability of vector infectivity.

Animals↗

Concentrations of human erythrocytes by anopheline mosquitoes (Diptera: Culicidae) during feeding.

Erythrocyte densities in the blood meals of six Anopheles mosquito species were compared with those of human host erythrocyte densities. During engorgement, An. gambiae Giles and An. stephensi Liston concentrated erythrocytes by factors of 1.8 and 1.7, respectively; An. freeborni Aitken did not concentrate; and An. arabiensis Patton and An. dirus Peyton & Harrison demonstrated an intermediate level of erythrocyte concentration (1.4 and 1.2, respectively). An. albimanus concentrated host hemoglobin, but hemolysis during engorgement decreased bloodmeal erythrocyte density below that of host blood. The degree to which anopheline species concentrated erythrocytes was related to the frequency and time spent undergoing prediuresis (anal excretion of fluid during feeding), suggesting that prediuresis is responsible for erythrocyte concentration and that the fluid produced represents efflux from the filtration of ingested blood. Differences observed in erythrocyte concentration by different anopheline species are consistent with species-specific patterns of host selection.

Animals↗

Quantitation of Plasmodium falciparum sporozoites transmitted in vitro by experimentally infected Anopheles gambiae and Anopheles stephensi.

The frequency and numbers of Plasmodium falciparum sporozoites transmitted in vitro and corresponding sporozoite loads were determined for experimentally infected Anopheles gambiae and An. stephensi. Geometric mean (GM) sporozoite loads in three experiments ranged from 808 to 13, 905 for An. gambiae and from 6, 608 to 17, 702 for An. stephensi. A total of 44.1% of 68 infected An. gambiae and 49.2% of 63 infected An. stephensi transmitted sporozoites in vitro. The GM number of sporozoites transmitted was 4.5 for An. gambiae and 5.4 for An. stephensi. Overall, 86.9% of the mosquitoes transmitted from one to 25 sporozoites, and only 6.6% transmitted over 100 sporozoites (maximum = 369). Sporozoite loads were not a useful predictor of potential sporozoite transmission. Despite higher sporozoite loads, the numbers of sporozoites transmitted in vitro by the experimentally infected mosquitoes were similar to estimates obtained, using the same techniques, for naturally infected An. gambiae in western Kenya. The low but highly variable numbers of sporozoites transmitted in vitro by mosquitoes used in malaria vaccine challenge studies appears to be a reasonable simulation of natural sporozoite transmission.

Animals↗

Flea-borne rickettsioses: ecologic considerations.

Ecologic and economic factors, as well as changes in human behavior, have resulted in the emergence of new and the reemergence of existing but forgotten infectious diseases during the past 20 years. Flea-borne disease organisms (e.g., Yersinia pestis, Rickettsia typhi, R. felis, and Bartonella henselae) are widely distributed throughout the world in endemic-disease foci, where components of the enzootic cycle are present. However, flea-borne diseases could reemerge in epidemic form because of changes in vector-host ecology due to environmental and human behavior modification. The changing ecology of murine typhus in southern California and Texas over the past 30 years is a good example of urban and suburban expansion affecting infectious disease outbreaks. In these areas, the classic rat-flea-rat cycle of R. typhi has been replaced by a peridomestic animal cycle involving, e.g., free-ranging cats, dogs, and opossums and their fleas. In addition to the vector-host components of the murine typhus cycle, we have uncovered a second typhuslike rickettsia, R. felis. This agent was identified from the blood of a hospitalized febrile patient and from opossums and their fleas. We reviewed the ecology of R. typhi and R. felis and present recent data relevant to the vector biology, immunology, and molecular characterization and phylogeny of flea-borne rickettsioses.

Animals↗