Aerobic Conditions and Endogenous Reactive Oxygen Species Reduce the Production of Infectious MS2 phage by Escherichia coli

Most of the defective/non-infectious enteric phages and viruses that end up in wastewater originate in human feces. Some of the causes of this high level of inactivity at the host stage are unknown. There is a significant gap between how enteric phages are environmentally transmitted and how we might design molecular tools that would only detect infectious ones. Thus, there is a need to explain the low proportion of infectious viral particles once replicated. By analyzing lysis plaque content, we were able to confirm that, under aerobic conditions, Escherichia coli produce low numbers of infectious MS2 phages (I) than the total number of phages indicated by the genome copies (G) with an I/G ratio of around 2%. Anaerobic conditions of replication and ROS inhibition increase the I/G ratio to 8 and 25%, respectively. These data cannot only be explained by variations in the total numbers of MS2 phages produced or in the metabolism of E. coli. We therefore suggest that oxidative damage impacts the molecular replication and assembly of MS2 phages.

The Effects of Temperature and Host Stage on Development Rate of Parasitoids of Giant Whitefly Aleurodicus dugesii (Hemiptera: Aleyrodidae)

The effects of rearing temperature and host stage parasitized on the development of three parasitoid species; Encarsia noyesi Hayat (Hymenoptera: Aphelinidae), Idioporus affinis LaSalle & Polaszek (Hymenoptera: Pteromalidae), and Entedononecremnus krauteri Zolnerowich & Rose (Hymenoptera: Eulophidae) were investigated. These parasitoids are part of the biological control program for the giant whitefly Aleurodicus dugesii Cockerell (Hemiptera: Aleyrodidae) in the United States. Temperature and host-dependent development was assessed for each species using the nonlinear Brière-1 model and simple linear regression to obtain critical thermal parameters. All three parasitoids successfully developed at constant temperatures ranging from 15°C to 30°C, which was narrower to their predicted thermal limits due to thermal constraints of A. dugesii. There were significant effects of both temperature and A. dugesii nymphal stage parasitized on immature parasitoid development times. All three parasitoid species’ development time decreased as nymphal stage age increased. Thermal tolerance limits and development times varied by parasitoid species. The results of these findings in the context of biological control potential are discussed.

INFLUENCE OF HOST STAGE ON OVIPOSITION, DEVELOPMENT, AND SEX RATIO OF Anagyrus lopezi (DE SANTIS) (HYMENOPTERA: ENCYRTIDAE), A PARASITOID OF THE CASSAVA MEALYBUG, Phenacoccus manihoti MATILE-FERRERO (HEMIPTERA: PSEUDOCOCCIDAE)

Influence of host stage on oviposition, development, and sex ratio of Anagyrus lopezi (De Santis) (Hymenoptera: Encyrtidae), a parasitoid of the cassava mealybug, Phenacoccus manihoti Matile-Ferrero (Hemiptera: Pseudococcidae). The parasitoid Anagyrus lopezi (De Santis) (Hymenoptera: Encyrtidae) was introduced from Thailand into Indonesia in early 2014 to control the invasive cassava mealybug, Phenacoccus manihoti Matile-Ferrero (Hemiptera: Pseudococcidae). Because of the need to produce large numbers of high-quality females, research was conducted in the laboratory to determine host stage preference for A. lopezi on different instars of P. manihoti. Individual female wasps were exposed to first, second, third instar nymphs, and pre-reproductive adult mealybugs. In the no-choice test, the frequency of parasitized hosts and the number of eggs laid per host was significantly higher in second and third instar nymphs as well as adult mealybugs compared to first instar nymphs. In the two-choice test, third instars nymphs and adult mealybugs were the most preferred host for oviposition. Immature development of parasitoids was faster and the ratio of female to male parasitoids was higher following oviposition in second and third instar nymphs and pre-reproductive adult hosts, compared to the first instar nymphs. Our findings indicate that the use of pre-reproductive adults as hosts in a mass-rearing program would be the most productive and fastest way to produce A. lopezi populations with a female-biased sex ratio. Field release of parasitoids should be conducted when the host’s third instar nymph is the most abundant because the period during which preferred and suitable host stages are available would be the longest.

Methodical Considerations and Resistance Evaluation against Fusarium graminearum and F. culmorum Head Blight in Wheat. Part 3. Susceptibility Window and Resistance Expression

Flowering is the most favorable host stage for Fusarium infection in wheat, which is called the susceptibility window (SW). It is not known how long it takes, how it changes in different resistance classes, nor how stable is the plant reaction in the SW. We have no information, how the traits disease index (DI), Fusarium-damaged kernel rate (FDK), and deoxynivalenol (DON) respond within the 16 days period. Seven winter wheat genotypes differing in resistance were tested (2013–2014). Four Fusarium isolates were used for inoculation at mid-anthesis, and 4, 8, 11, 13, and 16 days thereafter. The DI was not suitable to determine the length of the SW. In the Fusarium-damaged kernels (FDK), a sharp 50% decrease was found after the 8th day. The largest reduction (above 60%) was recorded for DON at each resistance level between the 8th and 11th day. This trait showed the SW most precisely. The SW is reasonably stable in the first 8–9 days. This fits for all resistance classes. The use of four isolates significantly improved the reliability and credit of the testing. The stable eight-day long SW helps to reduce the number of inoculations. The most important trait to determine the SW is the DON reaction and not the visual symptoms.

ASPECTS OF BIOLOGY OF Acerophagus papayae Noyes & Schauff (HYMENOPTERA: ENCYRTIDAE), PARASITOID OF THE PAPAYA MEALYBUG

Aspects of biology of Acerophagus papayae Noyes & Schauff (Hymenoptera: Encyrtidae), parasitoid of papaya mealybug. Acerophagus papayae Noyes & Schauff (Hymenoptera: Encyrtidae) is an important parasitoid of the papaya mealybug, Paracoccus marginatus Williams & Granara de Willink (Hemiptera: Pseudococcidae). The study was conducted with the objective to determine various aspects of the biology of A. papayae which include the effect of diet on adult longevity, fecundity and progeny, host stage susceptibility and preference, the effect of host stages on immature development, body size, and sex ratio of progenies. Effects of diet on adult longevity was done in the absence of hosts. Fecundity was measured by the number of mealybugs parasitized. Host stage susceptibility and preference were carried out by exposing 2nd and 3rd nymphal instars and pre-reproductive adults of mealybugs to parasitoids. Results showed adult parasitoids fed with 10% honey solution lived almost fourfold longer than those provided only water. A. papayae parasitized 30.1±4.92 mealybugs, with a range of 13-60 mealybugs, during 5.8 days of adult life. In no-choice (susceptibility) and paired-choice (preference) tests, the percentage of parasitized hosts were significantly greater in 2nd and 3rd instar nymphs than in adults. The mean immature developmental time of A. papayae was longer when the parasitoids develop in large host. Developmental time of male parasitoids was shorter than the females. Female wasps which emerged from hosts parasitized at the 3rd instar nymphs and adults were significantly larger than those from the 2nd instar nymphs. Sex ratios of the offspring emerged from hosts that were parasitized as 2nd instars were strongly male-biased, while the later stages yielded more females than males.

The evolution of stage-specific virulence: differential selection of parasites in juveniles

The impact of infectious disease is often very different in juveniles and adults, but theory has focused on the drivers of stage-dependent defense in hosts rather than the potential for stage-dependent virulence evolution. Stage-structure has the potential to be important to the evolution of pathogens because it exposes parasites to heterogeneous environments in terms of both host characteristics and transmission routes. We develop a stage-structured (juvenile-adult) epidemiological model and examine the evolutionary outcomes of stage-specific virulence under the classic assumption of a transmission-virulence trade-off. We show that selection on virulence against adults remains consistent with the classic theory. However, the evolution of juvenile virulence is sensitive to both demography and transmission pathway with higher virulence against juveniles being favored either when the transmission pathway is assortative (juveniles preferentially interact together) and the juvenile stage is short, or in contrast when the transmission pathway is disassortative and the juvenile stage is long. These results highlight the potentially profound effects of host stage-structure on determining parasite virulence in nature. This new perspective may have broad implications for both understanding and managing disease severity.Impact summaryUnderstanding the evolution of parasite virulence remains one of the most important questions in evolutionary ecology. Virulence is often very different in young and old hosts, but previous theory has presumed that these differences are attributed to adaptation in host defense rather than parasite adaptation. However, stage-structure within host populations can expose parasites to heterogeneous environments, which may lead to differential selection on parasite virulence (stage-specific virulence). Surprisingly, no study has investigated the effects of hosts’ stage-structure on the evolution of stage-specific virulence. We present a theoretical analysis to examine when selection can favor higher virulence against juveniles (juvenile-virulence) versus adults (adult-virulence). Our key result is that higher juvenile-virulence is selected for either when the transmission is assortative within age classes and maturation is slow, or when the transmission is disassortative (occurring predominantly between-classes) and maturation is relatively fast. These at first sight contrasting outcomes can be understood as adaptation to the exploitation of the more available host stage. Although the data on assortativity in infectious disease systems is limited, empirical studies for the virulence of Great Island Virus in guillemots (Uria aalge) and for salmon louse in pink salmon (Oncorhynchus gorbuscha) are consistent with our predictions. Our work provides testable predictions for stage-specific virulence and presents a novel mechanism that may explain variation in virulence in nature. There are also management implications for conservation, public health, vaccination programs, and farming to understanding the drivers of stage dependent virulence.