Virus-infected Females Research Articles

Differential proteomics profiling of the ova between healthy and Rice stripe virus-infected female insects of Laodelphax striatellus.

Rice stripe virus-infected females of the small brown planthopper (SBPH, Laodelphax striatellus) usually lay fewer eggs with a longer hatch period, low hatchability, malformation and retarded or defective development compared with healthy females. To explore the molecular mechanism of those phenomena, we analyzed the differential proteomics profiling of the ova between viruliferous and healthy female insects using an isobaric tag for relative and absolute quantitation (iTRAQ) approach. We obtained 147 differentially accumulated proteins: 98 (66.7%) proteins increased, but 49 (33.3%) decreased in the ova of the viruliferous females. RT-qPCR was used to verify the 12 differential expressed proteins from iTRAQ, finding that trends in the transcriptional change for the 12 genes were consistent with those at the proteomic level. Differentially expressed proteins that were associated with meiosis (serine/threonine-protein phosphatase 2B and cyclin B3) and mitosis (cyclin B3 and dynein heavy chain) in viruliferous ova may contribute to low hatchability and defective or retarded development. Alterations in the abundance of proteins involved in the respiratory chain and nutrition metabolism may affect embryonic development. Our study begins to explain macroscopical developmental phenomena and explore the mechanisms by which Rice stripe virus impacts the development of SBPH.

Effects of Salivary Gland Hypertrophy Virus on the Reproductive Behavior of the Housefly, Musca domestica

Pathological studies demonstrated that the salivary gland hypertrophy virus of houseflies (MdSGHV) shuts down reproduction in infected females. The mechanism that underlay the disruption of reproduction functioned on several levels. Females infected at the previtellogenic stage did not produce eggs, reflecting a block in the gonadotropic cycle. Sodium dodecyl sulfate-polyacrylamide gel electrophoresis and Western blot analysis of hemolymph samples demonstrated that MdSGHV infection reduced the levels of both the female-specific hexamerin and egg yolk proteins. Furthermore, reverse transcriptase quantitative real-time PCR data demonstrated that infection blocked hexamerin and yolk protein gene transcription. When females were allowed to develop eggs prior to infection (postvitellogenic stage), the outcome of mating attempts depended upon when mating took place. If egg-containing, virus-infected females were mated within 24 h of infection, they copulated and deposited a single batch of fertilized eggs. However, if mating was delayed for a longer period, the egg-containing females refused to copulate with healthy males. Both of these results suggested that a virus-induced signal influenced the central nervous system, shutting down female receptivity and egg production. All experiments demonstrated that MdSGHV-infected males did not display azoospermia and were fertile. Both healthy females mated with infected males, and the resulting F1 progeny were free of salivary gland hypertrophy symptoms, which suggests that the virus is not sexually or vertically transmitted.

A TWO-YEAR EVALUATION OF ELEVATED CANOPY TRAPPING FOR CULEX MOSQUITOES AND WEST NILE VIRUS IN AN OPERATIONAL SURVEILLANCE PROGRAM IN THE NORTHEASTERN UNITED STATES

The effectiveness of CO2-baited Centers for Disease Control and Prevention miniature light traps elevated in the tree canopy (approximately 7.6 m) was compared with light traps placed at ground level (approximately1.5 m) and grass-sod infused gravid traps for collecting Culex pipiens, Culex restuans, and Culex salinarius and detecting West Nile virus (WNV) activity in an operational surveillance program that encompassed 12 ecologically diverse sites in Connecticut in 2004 and 2005. More than twice as many Cx. pipiens were collected on average in light traps suspended in the tree canopy than in either light or gravid traps placed at ground level. This difference was generally restricted to those collection sites where markedly greater numbers of Cx. pipiens were collected with all trapping methods but was not associated with site-specific urbanization indices. Culex restuans was not preferentially attracted to light traps suspended in the tree canopy. No differences in the overall abundance of this species were recorded with either of the 2 trapping procedures, but both light traps were more effective than the gravid traps. Culex salinarius was significantly more attracted to ground-based light traps than traps suspended in the tree canopy, while gravid traps were ineffective at all sites regardless of the level of urbanization or any other specific land-use characteristic. CO2-baited light traps placed in the tree canopy were generally superior to ground-based light traps for detecting WNV in Cx. pipiens. West Nile virus-infected females were collected more regularly, and the frequency of infected pools was significantly greater. Twofold higher minimum field infection rates (maximum likelihood estimation [MLE] = 6.7 vs. 3.0 per 1,000 mosquitoes) were also recorded from canopy collections of this species, and virus was detected in canopy-collected females several weeks before it was detected in collections from light traps at ground level. We conclude that the use of CO2-baited light traps placed in the tree canopy for targeted trapping of Cx. pipiens and subsequent detection of WNV are likely to yield better overall results than light traps placed at ground level in this region of the northeastern United States. The virus isolation data obtained from Cx. pipiens collected in gravid traps compared favorably both temporally and spatially with results from canopy trap collections. There were no significant differences in the overall frequency of WNV-infected pools or MLEs for Cx. pipiens, but fewer total WNV isolations were made from Cx. pipiens collected in the gravid traps and virus was detected more infrequently. Results reaffirmed the utility of gravid traps as effective surveillance tools for detection of WNV in Cx. pipiens in the northeastern United States. However, findings also demonstrated that CO2-baited light traps placed in the tree canopy provided more consistent results where weekly detection of virus amplification is a critical objective. The comparative effectiveness of ground- and canopy-based light traps for detection of WNV-infected Cx. restuans and Cx. salinarius was inconclusive owing to the limited number of virus isolations that were made from these species during the 2 years of study. However, WNV virus isolations were made several weeks earlier and more frequently from Cx. restuans collected in traps placed in the canopy rather than at ground level in 2004. Results support the view that ground-based light traps are more effective for detection of WNV in Cx. salinarius.

Effect of La Crosse Virus Infection on Insemination Rates in Female <I>Aedes triseriatus</I> (Diptera: Culicidae)

Aedes triseriatus (Say) (Diptera: Culicidae) females orally infected with La Crosse virus after ingesting an infectious bloodmeal were compared for mating efficiency with females that ingested a noninfectious bloodmeal. After 14-d extrinsic incubation to allow for dissemination of the infection, all females were offered a second noninfectious bloodmeal and were placed in cages with age-matched males for 5 d. After 6 d, insemination rates were determined by detection of sperm in the spermathecae. Insemination rates of the La Crosse virus-infected females were significantly greater than in uninfected females.

Infection with the insect virus Hz-2v alters mating behavior and pheromone production in female Helicoverpa zea moths

The effect of Hz-2V virus infection on the reproductive physiology and behavior of infected Helicoverpa zea female moths was examined. In the absence of males, infected females exhibited calling behavior and called as often but for shorter periods on average than control females. As expected, control females mated with males for extend periods when they were present and did not call after mating, while virus-infected females made many frequent contacts with males and continued to call even after these contacts. Virus-infected females were found to produce five to seven times more pheromone than control females and attracted twice as many males as did control females in flight tunnel experiments. The ability of Hz-2V to alter the physiology and behavior of infected females observed here may serve to facilitate the transmission of virus in insect populations.

Infection with the insect virus Hz-2v alters mating behavior and pheromone production in female Helicoverpa zea moths

The effect of Hz-2V virus infection on the reproductive physiology and behavior of infected Helicoverpa zea female moths was examined. In the absence of males, infected females exhibited calling behavior and called as often but for shorter periods on average than control females. As expected, control females mated with males for extend periods when they were present and did not call after mating, while virus-infected females made many frequent contacts with males and continued to call even after these contacts. Virus-infected females were found to produce five to seven times more pheromone than control females and attracted twice as many males as did control females in flight tunnel experiments. The ability of Hz-2V to alter the physiology and behavior of infected females observed here may serve to facilitate the transmission of virus in insect populations.

Iridovirus in the root weevil Diaprepes abbreviatus

Invertebrate iridescent virus 6 (IIV6) was evaluated for mode of transmission and ability to cause infection in the root weevil, Diaprepes abbreviatus (L.). This is the first evidence of IIV6 infection in D. abbreviatus, which caused both patent and sub-lethal covert infections in both larvae and adults. Adults and larvae were successfully infected with IIV6 by puncture, injection and per os. Transmission of IIV6 was demonstrated between infected and healthy individuals regardless of gender. Virus was detected in egg masses produced by virus-infected females suggesting IIV6 is transmitted transovarially. Virus particles were observed in the cytoplasm of weevil cells, and were shown to infect fat bodies, muscle, and nerve tissues, as visualized using transmission electron microscopy. Patent infections resulted in death of individuals within 3 to 4 days post infection. Individuals with covert infections tested positive for virus infection on day 7 by polymerase chain reaction analysis. Sequencing of PCR amplicons confirmed virus infection. Discovery of new pathogens against root weevils may provide new management tools for development of control strategies based on induced epizootics. This is the first report of a virus infecting D. abbreviatus.

Iridovirus in the root weevil Diaprepes abbreviatus.

Invertebrate iridescent virus 6 (IIV6) was evaluated for mode of transmission and ability to cause infection in the root weevil, Diaprepes abbreviatus (L.). This is the first evidence of IIV6 infection in D. abbreviatus, which caused both patent and sub-lethal covert infections in both larvae and adults. Adults and larvae were successfully infected with IIV6 by puncture, injection and per os. Transmission of IIV6 was demonstrated between infected and healthy individuals regardless of gender. Virus was detected in egg masses produced by virus-infected females suggesting IIV6 is transmitted transovarially. Virus particles were observed in the cytoplasm of weevil cells, and were shown to infect fat bodies, muscle, and nerve tissues, as visualized using transmission electron microscopy. Patent infections resulted in death of individuals within 3 to 4 days post infection. Individuals with covert infections tested positive for virus infection on day 7 by polymerase chain reaction analysis. Sequencing of PCR amplicons confirmed virus infection. Discovery of new pathogens against root weevils may provide new management tools for development of control strategies based on induced epizootics. This is the first report of a virus infecting D. abbreviatus.

The effects of a DNA virus infection on the reproductive potential of female tsetse flies, Glossina morsitans centralis and Glossina morsitans morsitans (Diptera: Glossinidae).

Reproductive anomalies associated with the tsetse DNA virus infection in the female tsetse hosts, Glossina morsitans centralis Machado and Glossina morsitans morsitans Westwood, inoculated with the virus during the 3rd instar larval stage were studied and the data compared to those obtained from the control females injected with sterile physiological saline. Virus infected flies had significantly longer first and second pregnancy cycles (P < 0.0001) and produced pupae that were of significantly less weight in milligrams (P < 0.0001) compared to controls. Transmission of the virus to progeny was not absolute and only 21% of G. m. centralis and 48% of G. m. morsitans first progeny flies from infected females developed salivary gland hypertrophy as a result of transmission from mother to progeny. The virus infected females produced significantly fewere pupae compared to the controls during the experimental period (P < 0.00001).

Limiting conditions of the horizontal transmission of the Drosophila C virus in its host (D. melanogaster)

Abstract We carried out experiments with virus infected females on the horizontal transmission of Drosophila C virus on artificial culture medium and under controlled conditions. The numerical results made it possible to build up an empirical model. We tested the values of the rearing parameters under nearly natural conditions in boxes containing pieces of rotting apple. A good agreement between observed and estimated values led to the following conclusions: (i) The horizontal transmission of the Drosophila C virus is facilitated via the contamination of the nutritive medium by the cadavers or the feces of virus-infected females (ii) The contamination depends on the richness of viral particles of the contaminating flies. We also confirmed under ‘natural’ conditions the insufficient contamination of the male flies which justifies the use of females as ‘reservoirs’ of the virus. The estimation of the biological limiting conditions for horizontal transfer of the virus is the following: a minimum of two contaminating females have to stay for at least two days on an initially uninfected milieu so that a durable contamination of at least some females results. Under these conditions, the richness in viral particles in the contaminated flies is lower than that in the contaminating females and therefore the long term virus infection remains problematical.

Gonadal lesions in virus-infected male and female tsetse, Glossina pallidipes (Diptera: Glossinidae)

Ovaries and testes, dissected out and collected from wild Glossina pallidipes with virus-infected, hypertrophied salivary glands, were processed for routine electron microscopy and examined for pathological changes. Lesions in the male gonads were more severe than in their female counterpart and involved both testes in every case. Examination of the ovaries of the virus-infected females with hypertrophied salivary glands showed that although the occasional germarium appeared normal, the majority were affected by degeneration and severe necrosis. The degenerate ovarioles also showed virus particles within germarial cells. Sections of testicular follicles of adult males with hypertrophied salivary glands revealed a total arrest of spermatogenesis, depicted by complete absence of meiotic and postmeiotic stages. The follicles contained only sheets of undifferentiated premeiotic spermatogenic cells, i.e., spermatogonia and primary spermatocytes. In some testes, degeneration (characterized by diffuse vacuolation and exfoliation of degenerate cells leading to empty lumina) was superimposed on lack of sperm development. Testes of normal adult male G. pallidipes, on the other hand, showed swarms of entangled spermatozoa. The demonstration of virus particles within the germarium of the ovariole of females with hypertrophied salivary glands and the complete sterility observed in males with hypertrophied salivary glands identify the virus of G. pallidipes as a potential candidate for the biological control of the species.

Milk-borne transmission of human T-cell leukemia virus type I in rabbits

The mode of vertical transmission of human T-cell leukemia virus type I (HTLV-I) was investigated by foster-nursing in rabbits. Two of five rabbits born to noninfected mothers and fostered by virus-infected females seroconverted for HTLV-I after 7 weeks, whereas all seven rabbits born to virus-infected mothers and fostered by noninfected females remained seronegative for the observation period of 6 months. Culture of peripheral blood lymphocytes gave rise to an HTLV-I-carrying T-cell line from each of the two seroconverted rabbits. These findings suggest that HTLV-I is transmitted through milk from mother to offspring.

Mother-to-Offspring Transmission of Human T Cell Leukemia Virus Type I in Rabbits

We studied the mode of natural transmission of human T-cell leukemia virus type I (HTLV-I) in rabbits. Four virus-infected rabbits (2 males and 2 females) were individually mated with 4 noninfected rabbits. Two virus-infected females mated with noninfected males gave birth to 7 offspring, and 2 noninfected females mated with infected males delivered 5 offspring. Four of the seven offspring born to the virus-infected mothers seroconverted for HTLV-I when aged 6 to 13 weeks with antibody titers of 1:40 to 1:160. None of the five offspring born to the noninfected mothers became seropositive during the observation period of 6 months, however. Peripheral lymphocytes were cultured with T cell growth factor, and HTLV-I-carrying lymphoid cell lines were established from the four seroconverted rabbits. All four cell lines were of T cells positive for Ia antigens. In addition, none of five newborn rabbits killed immediately after birth to a virus-infected rabbit was infected with HTLV-I. These findings provide an experimental support for the milkborne transmission of HTLV-I from mother to child in humans and indicate that the virus is tropic for T cells in rabbits as well.

Mother-to-offspring transmission of human T cell leukemia virus type I in rabbits

We studied the mode of natural transmission of human T-cell leukemia virus type I (HTLV-I) in rabbits. Four virus-infected rabbits (2 males and 2 females) were individually mated with 4 noninfected rabbits. Two virus-infected females mated with noninfected males gave birth to 7 offspring, and 2 noninfected females mated with infected males delivered 5 offspring. Four of the seven offspring born to the virus- infected mothers seroconverted for HTLV-I when aged 6 to 13 weeks with antibody titers of 1:40 to 1:160. None of the five offspring born to the noninfected mothers became seropositive during the observation period of 6 months, however. Peripheral lymphocytes were cultured with T cell growth factor, and HTLV-I-carrying lymphoid cell lines were established from the four seroconverted rabbits. All four cell lines were of T cells positive for Ia antigens. In addition, none of five newborn rabbits killed immediately after birth to a virus-infected rabbit was infected with HTLV-I. These findings provide an experimental support for the milkborne transmission of HTLV-I from mother to child in humans and indicate that the virus is tropic for T cells in rabbits as well.

Transmission of a baculovirus in populations of Oryctes rhinoceros

The transmission of the baculovirus of Oryctes rhinoceros, previously called Rhabdionvirus oryctes, was studied. O. rhinoceros adults became infected with the virus when kept in a mixture of sawdust and ground-up virus-killed larvae or together with other virus-infected adults. In the field, mated females were more frequently infected than unmated females. Adults developing from larvae that had survived exposure to various dosages of the virus were not infected. No virus infections occurred in larvae hatching from eggs surface-contaminated with the virus. Larvae hatching from eggs laid by virus-infected females very rarely were infected. In the O. rhinoceros population the virus is transmitted most frequently during mating, possibly when the uninfected partner contacts virus material excreted by the infected partner. The virus can be transmitted in a similar way when infected and healthy beetles feed together in palm trees. Beetles visiting larval breeding sites containing freshly virus-killed larvae can become infected. Virus-infected beetles can pass the infection to healthy larvae when visiting a breeding site.