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Daily oviposition patterns of the African malaria mosquito Anopheles gambiae Giles (Diptera: Culicidae) on different types of aqueous substrates.

BACKGROUND: Anopheles gambiae Giles is the most important vector of human malaria in sub-Saharan Africa. Knowledge of the factors that influence its daily oviposition pattern is crucial if field interventions targeting gravid females are to be successful. This laboratory study investigated the effect of oviposition substrate and time of blood feeding on daily oviposition patterns of An. gambiae mosquitoes. METHODS: Greenhouse-reared gravid and hypergravid (delayed oviposition onset) An. gambiae sensu stricto and wild-caught An. gambiae sensu lato were exposed to three types of substrates in choice and no-choice cage bioassays: water from a predominantly anopheline colonised ground pool (anopheline habitat water), swamp water mainly colonised by culicine larvae (culicine habitat water) and distilled water. The daily oviposition pattern and the number of eggs oviposited on each substrate during the entire egg-laying period were determined. The results were subjected to analysis of variance using the General Linear Model (GLM) procedure. RESULTS: The main oviposition time for greenhouse-reared An. gambiae s.s. was between 19:00 and 20:00 hrs, approximately one hour after sunset. Wild-caught gravid An. gambiae s.l. displayed two distinct peak oviposition times between 19:00 and 20:00 hrs and between 22:00 and 23:00 hrs, respectively. During these times, both greenhouse-reared and wild-caught mosquitoes significantly (P < 0.05) preferred anopheline habitat water to the culicine one. Peak oviposition activity was not delayed when the mosquitoes were exposed to the less preferred oviposition substrate (culicine habitat water). However, culicine water influenced negatively (P < 0.05) not only the number of eggs oviposited by the mosquitoes during peak oviposition time but also the overall number of gravid mosquitoes that laid their eggs on it. The differences in mosquito feeding times did not affect the daily oviposition patterns displayed. CONCLUSION: This study shows that the peak oviposition time of An. gambiae s.l. may be regulated by the light-dark cycle rather than oviposition habitat characteristics or feeding times. However, the number of eggs laid by the female mosquito during the peak oviposition time is affected by the suitability of the habitat.

Journal Article↗

Collateral uterine blood flow in Holstein cows during the third trimester of pregnancy.

Blood flow to the gravid and nongravid uterine horns of four multiparous Holstein cows (mean +/- SD, BW=641.8 +/- 95.4 kg; age=4.8 +/- 1.2 years; parity=3.0 +/- 1.2) was measured on days 225, 248, and 266 of gestation. Surgery was conducted on day 214.5 +/- 4.0 of gestation through the flank of the standing cows. Transit-time ultrasonic flow probes (diameter 12 or 14 mm) were fitted surgically around the uterine arteries of each cow. Surgery was completed within two hours of anesthesia, and the animals recovered rapidly following surgery. Uterine blood flow (UBF, l/min) was recorded at 10 sec intervals for approximately 23.5 hours; these values were averaged to determine UBF. The mean gravid UBF was significantly (P<0.05) greater than the nongravid UBF in this study. The range of the gravid and nongravid UBFs varied from 3.61 to 14.05 and 0.72 to 6.54 l/min, respectively. There were no changes (P>0.1) in the mean gravid and nongravid UBFs from day 225 to 266 of gestation.

Animals↗

Influence of latent 'asymptomatic' toxoplasmosis on body weight of pregnant women.

The latent toxoplasmosis is usually considered to be asymptomatic, however, this paradigm has never been rigorously tested. Here we searched for symptoms of deterioration of physical health (decrease of weight) in infected people by analysis of clinical records of 758 women tested for toxoplasmosis in the 16th week of gravidity. Toxoplasma-positive women have a lower body weight in the 16th week of gravidity (p = 0.02) than Toxoplasma-negative women. Moreover, a negative correlation between weight and the duration of toxoplasmosis was found in a subset of 174 Toxoplasma-positive women (p = 0.04), suggesting that slow and cumulative effects of latent toxoplasmosis, rather than a transient effect of acute toxoplasmosis, are responsible for the decreased weight of infected subjects. Longer duration of gravidity estimated from the date of last menstruation in the set of Toxoplasma-positive women in the 16th week of gravidity estimated with ultrasonography (p = 0.04) suggests a possibility of retarded foetal growth in Toxoplasma-positive women. The prevalence of latent toxoplasmosis is extremely high. Therefore, even its mild symptoms such as the decreased body weight in Toxoplasma-positive pregnant women might in fact indicate an unrecognized serious public health problem.

Adult↗

Effect of the conceptus on quantitative and qualitative aspects of uterine secretion in pigs.

Endometrial explants were removed from uteri of animals pregnant and pseudopregnant (5 mg oestradiol valerate on Days 11--15) for 60 days, from the gravid and non-gravid horns of unilaterally pregnant pigs at Day 60 of pregnancy and fron non-pregnant animals at Day 12 of the oestrous cycle. The tissues were cultured in the presence of L-[34S]methionine for 24 h, and the tissues and medium were then analysed separately by two-dimensional polyacrylamide gel electrophoresis. Radioactive polypeptides were identified by autoradiography of dried gels. Tissues from all all except the cyclic animals released an identical group of polypeptides into the culture medium: the major radioactive products were 4 acidic polypeptides of low molecular weight and several basic proteins, which included the purple phosphatase uteroferrin, and lysozyme. In separate experiments explants from 3 unilaterally pregnant pigs were cultured with L-[3H]leucine, and, on a fresh weight basis, the tissue from the non-gravid horn released significantly less radioactive macromolecular material into the medium in 24 h than did tissue from gravid horns. It therefore appears that although the nature of the secretion produced by the pregnant uterus is a consequence of maternal hormonal regulation alone, the tissue underlying a conceptus is quantitatively more active than that from unoccupied regions of a uterus.

Acid Phosphatase↗

Correlation between the release of ovine trophoblast protein-1 by the conceptus and the production of polypeptides by the maternal endometrium of ewes.

We studied the biosynthesis of two proteins, p70 (Mr 70,000; pI 4.0) and p15 (Mr 15,000; pI 5.7), by endometrial tissues from ewes between Days 12 and 24 of pregnancy and between Days 12 and 16 of the oestrous cycle to determine whether production of the two was correlated with the period of biosynthesis of ovine trophoblast protein-1 (oTP-1) by the conceptus. We also compared the protein synthetic activities of endometrium from gravid and non-gravid horns of pregnant ewes at Days 14, 16 and 18 in which the conceptus had been confined to one uterine horn. Proteins p70 and p15 were produced maximally between Days 14 and 20 of pregnancy, but synthesis by endometrial cultures from cyclic ewes was low or absent. Furthermore, synthesis of Protein p70 in particular was much greater by the gravid than non-gravid horn of unilaterally pregnant ewes. We conclude that synthesis of Proteins p70 and p15 by the uterus of sheep coincides with the time of oTP-1 production by the conceptus.

Animals↗

Significance of serum early pregnancy factor concentrations during pregnancy and embryonic development in Sminthopsis macroura (Spencer) (Marsupialia: Dasyuridae).

Marsupial pregnancy differs from that in eutherians in duration, placentation and hormonal profile so much so that maternal recognition of pregnancy may not occur in polyovular marsupials. However, a comparison of gravid and non-gravid uteri reveals differences indicative of histological and physiological adaptations to pregnancy. In the present study, the hypothesis that embryo-maternal signalling occurs in polyovular marsupials was tested by examining serum from non-pregnant and pregnant Sminthopsis macroura for the presence of early pregnancy factor (EPF), a serum protein secreted by the ovary in response to the presence of a newly fertilized egg in the oviduct. EPF is detectable in the serum of pregnant, but not in non-pregnant, females in all eutherians studied to date. In the present study, EPF was detected in S. macroura serum by the rosette inhibition test during the first 9 days of the 10.7 day gestation period in this marsupial. However, EPF was not detected on day 10, just before parturition, or in non-pregnant or preovulatory animals. Immunohistochemical analysis of ovaries from gravid and non-gravid animals demonstrates that EPF is found in the capillaries, interstitial spaces and secretory cells of the corpus luteum. It is concluded that the spatiotemporal pattern of EPF activity described strongly indicates that maternal recognition of pregnancy in marsupials is mediated, at least in part, by EPF. Because the endocrinological milieu is the same in pregnant and non-pregnant marsupials, the possibility of using marsupials as an experimental system for studying EPF function unconfounded by hormonal effects is presented.

Animals↗

Localization of transforming growth factor beta2 (TGFbeta2) and its receptors, TbetaRI and TbetaRII, in uteri and blastocysts of the stripe-faced dunnart (Sminthopsis macroura) during gastrulation.

This study describes for the first time the spatial and temporal distribution of a growth factor and its receptors in uteri and conceptuses of a marsupial species during the peri-gastrulation period. Uteri (gravid and non-gravid) and blastocysts from 40 female stripe-faced dunnarts (Sminthopsis macroura) were collected over the peri-gastrula period (days 6.0-8.5) and stained immunohistochemically for transforming growth factor beta2 (TGFbeta2) and its receptors, TbetaRI and TbetaRII, to determine possible roles for TGFbeta2 in marsupial embryonic development. The events that occurred during the period examined included the appearance and proliferation of hypoblast and mesoderm, primitive streak and node formation, and early neurulation. Differences in TGFbeta2 quantities between gravid and non-gravid uteri reflect differences in uterine morphology, indicating a role for TGFbeta2 in endometrial remodelling. In blastocysts, large quantities of all three proteins in the trophectoderm during the node stage coincide with both blastocyst expansion before implantation and the appearance of multinucleated cells, indicating that TGFbeta2 plays a role in conceptus elongation and trophectoderm differentiation. In contrast, lack of TbetaRII in blastocysts during hypoblast formation and proliferation negates any role for TGFbeta2 in these processes, as both receptors are required for a response to TGFbeta2. High concentrations of TGFbeta2 but low concentrations of TbetaRII in blastocysts during early primitive streak formation indicate that paracrinal embryo-maternal signalling may be occurring, as blastocysts cannot respond to TGFbeta2 at this stage, but uteri could. A similar situation, but reversed, also occurs during primitive node formation.

Activin Receptors, Type I↗

Differential expression of mesotocin receptors in the uterus and ovary of the pregnant tammar wallaby.

Mesotocin, an oxytocin-like peptide, is released in highest concentrations during parturition in macropodid marsupials. In late pregnant wallabies, uterine sensitivity to mesotocin increases markedly in the myometrium of the gravid uterus. This coincides with a significant increase in myometrial mesotocin receptor concentrations 3-4 days before term. To date, there is no information on mesotocin receptor gene expression in female wallaby reproductive tissues. This study aimed to examine mesotocin receptor gene expression in the uterus and ovaries of pregnant tammar wallabies, and to localise mesotocin receptors within the uterus. An RT-PCR strategy produced a consensus nucleotide sequence of 834 bp, which encoded 278 amino acids of transmembrane domains I to VI. This protein sequence has approximately 80% homology with the bovine and rat oxytocin receptor exon 2 region. Only one mesotocin receptor was detected in the tammar genome. The myometrium and mammary gland both expressed a 4.1 kb mesotocin receptor gene transcript. Myometrial mesotocin receptor gene expression increased on day 22 of the 26-day gestation and was significantly higher in the gravid than the non-gravid uterus in late pregnancy. This pattern of mesotocin receptor gene expression paralleled mesotocin receptor concentrations. Mesotocin binding sites were localised only to the myometrium, the highest densities being observed in the gravid uterus. Finally, this study showed high expression of mesotocin receptors in the corpus luteum. The pattern of luteal mesotocin receptor expression differed from the myometrium, with a decrease in mesotocin receptors occurring on the day of expected births.

Amino Acid Sequence↗

Ca2+ sensitivity of skinned uterine muscle fibers in unilaterally pregnant rats.

We prepared chemically skinned uterine muscle fibers from both horns of unilaterally pregnant rats and investigated the effect of gestation on the Ca2+ sensitivity of the Ca(2)-induced contraction. The Ca2+ sensitivity was defined as the pCa (minus logarithm of free calcium concentration) required for half maximum activation of force production. Gravid horns did not show any statistical difference in the Ca2+ sensitivity between normal pregnant and unilaterally pregnant rats. Non-gravid horns of unilaterally pregnant rats before Day 14 of gestation showed the same behavior in the Ca2+ sensitivity as horns of normal pregnant rats. After Day 15 of gestation the Ca2+ sensitivity of non-gravid horn also increased but significantly less than that of gravid horn. Taking into account the hormonal situation, the present results suggest that some local factors, such as an increased size of conceptus (fetus and placenta) and/or intercellular binding conditions of myometrium modulated by the gap junction formation and the appearance of connexin molecules, may also be requisite for the regulation of the contractile activity associated with parturition and delivery of the fetus and placenta.

Animals↗

Population dynamics of sand flies (diptera: psychodidae) at two foci of leishmaniasis in Texas.

Sand flies were collected at a focus of leishmaniasis in Medina County, TX, from April through October 1997 and at a focus in Bexar County, TX, from April 1998 through December 1999. Lutzomyia diabolica (Hall) were collected from April through November with peak abundance in July. The male:female ratio of Lu. diabolica was 1:6.2. Almost all female Lu. diabolica in the collections were unfed. One gravid Lu. diabolica contained 49 ova. Female Lu. anthophora (Addis) were active from February through December with three peaks in abundance suggestive of successive generations. Unfed and gravid Lu. anthophora were collected in about equal numbers. Gravid females contained from 1 to 64 ova per female. The male:female ratio was 1:1.8, with male Lu. anthophora collected in all months. One female Lu. anthophora was found infected with Leishmania in July 1999. Lutzomyia texana (Dampf) were collected from April through October with peak abundance in April during 1997. The male:female ratio was 1:1.4, with most females unfed. Two gravid Lu. texana contained 32 and 102 ova. An undescribed species of Lutzomyia was found only at the Medina County site from May through September 1997. Trapping sites four times per month versus two times per month in 1999 did not appear to adversely affect the abundance of Lu. diabolica or Lu. anthophora. There were marked differences in the species composition and relative abundance at the different sites, indicating that the spatial distribution of sand flies is patchy in nature.

Animals↗

Responses of fertile and sterile screwworm (Diptera: Calliphoridae) flies to bovine blood inoculated with bacteria originating from screwworm-infested animal wounds.

ABSTRACT A simple bioassay system was developed to study locomotory and ovipositional responses of screwworm, Cochliomyia hominivorax (Coquerel), flies to bovine blood inoculated with eight species of coliform bacteria that were isolated from screwworm-infested animal wounds. When exposed to odors from bacteria-inoculated blood which was incubated for 72 h at 37 degrees C, approximately 50% of 7- and 10-d-old gravid females landed on the blood by the end of 15 min test exposure. Only 17% of 7-d-old reproductively sterile females (from irradiated pupae) with previtellogenic ovaries and 2% of 4-d-old vitellogenic females responded to the same treatment. Females generally reacted in greatest numbers to bacteria-inoculated blood incubated for 72 h, followed by 48 h, then 24 and 96 h. Males of all ages tested were unresponsive. Although oviposition occurred in tests with gravid females lasting for 1 h, with both inoculated blood and an uninoculated control, the inoculated sample was significantly better than the control at 48, 72, and 96 h incubation duration. Our results are consistent with the conclusion that the inoculated blood, when incubated for 48-72 h, gives off volatile chemicals which attract gravid females and contains an oviposition stimulant that acts following contact and feeding. The volatiles, once isolated and identified, may be useful for sampling gravid females in the field as well as improving the oviposition system in the mass-production facility of the screwworm eradication program.

Animals↗

Oestrogen metabolism in the endometrium, corpus luteum and ovarian residual tissue of the rabbit.

Reproductive tissues (uterine endometrium, corpus luteum and ovarian residual tissues) from pregnant and pseudopregnant rabbits were incubated with equimolar concentrations of [3H]oestrone and [3H]oestrone sulphate (0.375 pmol) to monitor the changes in oestrogen metabolism during the early stages of pregnancy (days 0, and 3-8 post coitum) and to investigate the embyonic effect upon maternal oestrogen metabolism. Oestradiol-17 beta was the major metabolite formed from oestrone and sulphoconjugation occurred in all tissues studied. Oestrone sulphate was converted primarily to oestradiol-17 beta-3-monosulphate. Endometrial 17 beta-oxidoreductase significantly decreased and sulphotransferase increased in activity during the preimplantation period, but no differences were noted between gravid and non-gravid horns in unilaterally pregnant animals, nor between pregnant or pseudopregnant animals. Significant decreases occurred in 17 beta-oxidoreductase and sulphotransferase activity in luteal tissue, but these were more than offset by increases in tissue weight. No differences in the activities in luteal tissues were detected between pregnant or pseudopregnant animals, nor between ovarian tissue adjacent to gravid or non-gravid uterine horns. The results show that significant changes occur in oestrogen metabolism in the rabbit endometrium and corpus luteum within 8 days after ovulation, and that these changes result from maternal factors expressed systemically rather than by the effects of the developing conceptus expressed locally.

Animals↗

Ookinete rates in Afrotropical anopheline mosquitoes as a measure of human malaria infectiousness.

Anopheles gambiae s.1. and An. funestus were sampled for Plasmodium spp. ookinetes in two P. falciparum-endemic sites in western Kenya. Since the ookinete is a transitional stage of short duration, occurring after fertilization and before oocyst development, only females in the half-gravid and gravid stages of blood digestion were examined. Preparations of homogenized midguts were spotted onto microslides and examined microscopically after staining with Giemsa. Overall, ookinetes were detected in 4.4% of 1,079 anophelines examined over an eight-month period. Anopheles funestus had higher ookinete rates than An. gambiae s.1., and ookinete rates were higher in half-gravid than in gravid An. gambiae s.1. Geometric mean numbers of ookinetes per infected female were less than five for each species at the two sites, and the maximum number observed was only 12. The low frequencies and numbers of ookinetes were sufficient to produce sporozoite rates of 4-18% in the vector populations. The intense transmission of P. falciparum in these two sites is maintained by anthropophilic vectors where only one in 23 blood meals initiates an infection of generally less than five ookinetes. Relationships between human malaria infectiousness and vector infectivity are dependent upon the high efficiency of the developmental transition from the ookinete to the subsequent oocyst and sporozoite stages.

Animals↗

Detection of falciparum malarial forms in naturally infected anophelines in Cameroon using a fluorescent anti-25-kD monoclonal antibody.

Anopheles gambiae s.s. and An. funestus were sampled in houses located in a Plasmodium falciparum-holoendemic site in southern Cameroon. The midguts of female mosquitoes in half-gravid or gravid stages of blood digestion were incubated with a fluorescent monoclonal antibody directed against the P. falciparum zygote/ookinete surface protein Pfs25 and examined using a fluorescent light microscope. Malarial forms were detected in 11.6% of the half-gravid mosquitoes and in 0.0% of the gravid ones (P = 0.012). No difference in infections or the occurrence of malarial forms between An. gambiae and An. funestus was observed. Overall, 127 malarial forms were counted and distributed among round forms, retorts, and ookinetes in 77.2%, 9.5%, and 13.4%, respectively. Round forms include macrogametes, activating microgametocytes, and zygotes. The mean number of malarial forms per infected midgut was 2.16 and the maximum number observed was 13. In four anophelines, round forms, retorts, and ookinetes were simultaneously observed. Sporozoite rates were 5.7% for An. gambiae and 3.8% for An. funestus. In the human population, the gametocyte index for P. falciparum was 38% with a mean density of 1.11 gametocytes per microliter of blood. Differences concerning malarial forms in mosquito midguts were observed between houses (range percentage = 4.7--21.3%; mean range of forms per positive anopheline = 1.1--3.1). In each house, relationships existed between infected vectors and the gametocyte reservoir of their inhabitants. The role in transmission of people with very low gametocytemia, approximately one per microliter, as a reservoir of falciparum malaria in highly endemic areas, is emphasized.

Adolescent↗

Comparative effectiveness of three adult mosquito sampling methods in habitats representative of four different biomes of California.

The effectiveness of New Jersey (NJ) light, dry ice baited, and gravid female traps for collecting adult mosquitoes was compared at representative habitats in the Coachella, San Joaquin, and Sacramento valleys and the Los Angeles basin of California. The NJ light traps effectively sampled Anopheles freeborni, Culex tarsalis, Psorophora columbiae, and several Aedes when abundance was high in rural areas with minimal competitive illumination. Dry ice-baited encephalitis virus surveillance or CDC style traps collected significantly more females of most species at most localities than did NJ light traps, regardless of background illumination. The Cummings modification of the Reiter gravid female trap baited with a bulrush (Schoenoplectus) infusion was the best method for collecting Culex pipiens complex females in most habitats. In the Los Angeles basin, gravid traps baited with bulrush infusion collected, on average, 4.5 times more Culex quinquefasciatus females than did traps baited with the Reiter infusion. The bulrush infusion in combination with the Cummings trap design seemed to provide resting site cues and collected males as well as empty and bloodfed females. Mosquito surveillance programs in California should include the systematic operation of dry ice-baited and gravid female traps to improve surveillance sensitivity for selected species in appropriate habitats.

Animals↗

West Nile virus in mosquitoes of Northern Ohio, 2001-2002.

From June 14 to August 30, 2001 and June 10 to August 22, 2002, the mosquitoes of Oberlin, Ohio were surveyed and tested for West Nile virus (WNV). Mosquitoes were trapped weekly using gravid traps and CO(2)-baited Centers for Disease Control light traps at seven sites in 2001 and eight sites in 2002 in woodlots within the city limits. A total of 12,151 mosquitoes, representing 14 species and one species group, were collected in gravid traps, with Culex pipiens/restuans being the most abundant, followed by Ochlerotatus triseriatus. In light traps, 12,510 mosquitoes were collected, with Aedes vexans being the most abundant, followed by Culex pipiens/restuans. All gravid trap collections were tested for WNV via reverse transcriptase-polymerase chain reaction. West Nile virus was first detected in July 2002. Of all species tested, the Cx. pipiens/restuans species group exhibited the highest minimum infection rate (MIR) and the greatest percentage of positive pools. Both the MIR and percent of positive pools increased significantly with the date, although abundance of Cx. pipiens/restuans in gravid trap collections did not.

Animals↗

Differential preferences of oviposition by Aedes mosquitos in man-made containers under field conditions.

A study was made of the oviposit behavior of gravid female Aedes mosquitos in man-made habitats under field conditions. The study showed that the gravid female Aedes mosquitos preferred containers with relatively easy access but not too open to external environmental influence. The dark surface of the containers served as the initial and long-range attractant to the breeding sites. Volatile chemicals generated by the decaying vegetation in the container may serve as a close-range attractant. Finally, the water quality and the quantity of 'food' derived from decaying vegetative matter in the water determined the amount of eggs deposited in each container. The study confirmed previous findings that each gravid female Aedes mosquito had the tendency to lay her eggs in more than one container. However, the results of the study suggests that under favorable conditions, each gravid female Aedes mosquito could be encouraged to lay all her eggs in a single breeding site.

Aedes↗

Distribution, behavior, and course of patency of Dracunculus insignis in experimentally infected ferrets.

From 106 ferrets experimentally infected with Dracunculus insignis, 273 female worms and 42 male worms were recovered. The percentage of female worms that mated, the location of all worms, and the rate of emergence of gravid females were recorded. Of 174 mature female worms recovered, only 11% had emerged before necropsy. Eighty-one percent of the male worms and 87% of all unmated (immature) females were found on the trunk of the animal or on the skin overlying it. Gravid female worms were found on the extremities (legs) 88% of the time. Almost one-third of gravid female worms were found on the left front leg. On 4 occasions, females were found in unusual locations: the tail, the cheek, the lumbar region of the spinal cord, and the peritoneal cavity associated with the omentum. The results showed that only a small percentage of mature female worms emerge, which suggests that infected persons harbor many more gravid female worms than indicated by reports of emergent worms. More than one-third of female worms were not mated, but it is likely that these immature females would be capable of further development. Furthermore, male worms have been observed to survive for up to 330 days, suggesting that male or unmated female worms may carry over from one transmission season to the next.

Animals↗