Mary J. Camp

Methyl 2,4,6-decatrienoates attract Stink bugs and tachinid parasitoids.

Halyomorpha halys (Stål) (Pentatomidae), called the brown marmorated stink bug (BMSB), is a newly invasive species in the eastern USA that is rapidly spreading from the original point of establishment in Allentown, PA. In its native range, the BMSB is reportedly attracted to methyl (E,E,Z)-2,4,6-decatrienoate, the male-produced pheromone of another pentatomid common in eastern Asia, Plautia stali Scott. In North America, Thyanta spp. are the only pentatomids known to produce methyl 2,4,6-decatrienoate [the (E,Z,Z)-isomer] as part of their pheromones. Methyl 2,4,6-decatrienoates were field-tested in Maryland to monitor the spread of the BMSB and to explore the possibility that Thyanta spp. are an alternate host for parasitic tachinid flies that use stink bug pheromones as host-finding kairomones. Here we report the first captures of adult and nymph BMSBs in traps baited with methyl (E,E,Z)-2,4,6-decatrienoate in central Maryland and present data verifying that the tachinid, Euclytia flava (Townsend), exploits methyl (E,Z,Z)-2,4,6-decatrienoate as a kairomone. We also report the unexpected finding that various isomers of methyl 2,4,6-decatrienoate attract Acrosternum hilare (Say), although this bug apparently does not produce methyl decatrienoates. Other stink bugs and tachinids native to North America were also attracted to methyl 2,4,6-decatrienoates. These data indicate there are Heteroptera in North America in addition to Thyanta spp. that probably use methyl 2,4,6-decatrienoates as pheromones. The evidence that some pentatomids exploit the pheromones of other true bugs as kairomones to find food or to congregate as a passive defense against tachinid parasitism is discussed.

Establishment of a bovine blastocyst-derived cell line collection for the comparative analysis of embryos created in vivo and by in vitro fertilization, somatic cell nuclear transfer, or parthenogenetic activation

Tools and methods for analyzing differences in embryos resulting from somatic cell nuclear transfer (NT) in comparison to those derived from normal fertilization are needed to define better the nature of the nuclear reprogramming that occurs after NT. To this end, a collection of bovine blastocyst-derived cell lines was created. In vitro expanded or hatched blastocysts, used as primary culture tissue, were from NT; in vitro maturation, fertilization, and culture (IVF); or parthenogenetic (P) activation. Also, five in vivo-fertilized and developed blastocysts were collected by uterine flushing on the eighth d postfertilization. Whole blastocysts were physically attached to STO feeder layers to initiate all of the cell lines generated. The majority of the cell lines in the collection are trophectoderm, 38 NT-derived, 6 in vivo-derived, 20 IVF-derived, and 13 P-derived. Trophectoderm identity was ascertained by morphology and, in many cases, interferon-tau production. Several visceral endoderm cell lines and putative parietal endoderm cell lines were also established. At approximately 5% efficiency, epiblast masses from NT and IVF blastocysts survived and were isolated in culture. Two epiblast masses were also isolated from P blastocysts. Spontaneous differentiation from the epiblast outgrowths resulted in the establishment of fibroblast cell lines. The use of the trophectoderm cell lines as a comparative in vitro model of bovine trophectoderm and placental function is discussed in relation to NT reprogramming.

Triple-acting Lytic Enzyme Treatment of Drug-Resistant and Intracellular Staphylococcus aureus

Multi-drug resistant bacteria are a persistent problem in modern health care, food safety and animal health. There is a need for new antimicrobials to replace over used conventional antibiotics. Here we describe engineered triple-acting staphylolytic peptidoglycan hydrolases wherein three unique antimicrobial activities from two parental proteins are combined into a single fusion protein. This effectively reduces the incidence of resistant strain development. The fusion protein reduced colonization by Staphylococcus aureus in a rat nasal colonization model, surpassing the efficacy of either parental protein. Modification of a triple-acting lytic construct with a protein transduction domain significantly enhanced both biofilm eradication and the ability to kill intracellular S. aureus as demonstrated in cultured mammary epithelial cells and in a mouse model of staphylococcal mastitis. Interestingly, the protein transduction domain was not necessary for reducing the intracellular pathogens in cultured osteoblasts or in two mouse models of osteomyelitis, highlighting the vagaries of exactly how protein transduction domains facilitate protein uptake. Bacterial cell wall degrading enzyme antimicrobials can be engineered to enhance their value as potent therapeutics.

Temperature Ranges, Growth Optima, and Growth Rates of Spiroplasma (Spiroplasmataceae, class Mollicutes) Species

Abstract. A new method was developed for determination of the doubling times of spiroplasmas. In this procedure, the time required for medium acidification of tubes in tenfold dilution series was recorded. Sixty-four spiroplasma strains, representing 24 groups and 11 subgroups, were studied. Eight strains representing putative new groups were also included in the study. Doubling times at 5, 10, 15, 20, 25, 30, 32, 37, 41, and 43°C were determined. The range of temperatures for spiroplasma growth was 5°–41°C. Twenty-three spiroplasmas had optima of 30°C, 29 had optima of 32°C, and 13 had optima of 37°C. The fastest growing spiroplasma was the MQ-4 strain (group XI), with a doubling time at optimal temperature of 0.6 h. The slowest was the Jamaican corn stunt strain B655 (subgroup I-3), with an optimal doubling time of 36.7 h. Spiroplasma strain B31 (group IV) had the widest range (5°–41°C), while the DW-1 strain and some subgroup I-3 strains had the narrowest, growing only at 25° and 30°C. Some spiroplasmas grew well at 41°C, but none grew at 43°C. The ability of spiroplasmas to withstand a wide range of temperatures may reflect the conditions to which they are exposed in nature, including the temperatures of the insect, tick, and/or plant hosts in which they are carried and the plant surfaces from which they may be acquired by arthropods.

Stimulatory male volatiles for the Neotropical brown stink bug, Euschistus heros (F.) (Heteroptera: Pentatomidae)

Volatiles compounds collected from the male Neotropical brown stink bug, Euschistus heros (F.), a serious Central and South American soybean pest, have been reevaluated. The proportion of three methyl esters is found to be quite different from previous study. A new blend is proposed as the male-produced stimulatory volatiles based on gas chromatographic-electroantennographic detection (GC-EAD) techniques. The three GC-EAD-active components reproducibly found in volatiles collected from males are methyl (2E,4Z)-decadienoate (53%), methyl 2,6,10-trimethyldodecanoate (3%), and methyl 2,6,10-trimethyltridecanoate (44%) respectively. Males of this hemipteran species needed to reach an age of ~15 days to produce enough male-specific compounds to be detected by capillary column GC equipped with flame ionization detector (FID). The amounts of these three stimulatory volatiles increased with age and appeared to reach a maximum at ~35 days old, with the release rate of ~2.5 mg/day/male and the ratio of these three components seemed not to be affected by aging.

Adult diapause morph of the brown stink bug, Euschistus servus (Say) (Heteroptera: Pentatomidae)

The brown stink bug, Euschistus servus (Say), is abundant throughout most of eastern North America and is commonly found feeding on soybean, mullein, beans, tomatoes, peas, cotton, wheat, corn, tobacco and peach. Color change in E. servus from green to reddish-brown was shown to be an indicator of reproductive diapause. Reddish-brown insects lived longer than green individuals, females laid no eggs, and males did not produce pheromone. The high mortality registered for the green colony of E. servus adults was associated with the physiological cost associated with reproduction. The main pheromone component of this species is methyl 2E,4Z-decadienoate, in agreement with previous work. The first generation of this species develops on noncrop hosts and the second generation often migrates to crops where they may then exceed economic damage thresholds. Traps or trap crops baited with pheromone to catch or concentrate females for destruction, or even a pheromone-based disruption of orientation behavior to decrease the mating success, are possible semiochemical techniques to suppress populations of second generation of E. servus.

α-Glucosidase inhibitory activity and antioxidant capacity in the peel and pulp of mixed-species blueberry hybrids

Inhibition of α-glucosidase activity is considered an effective means for controlling diabetes by regulating glucose uptake, and blueberries have been shown to possess high levels of inhibitory activity. In the present study, we examined the variations in α-glucosidase inhibition, phenolic and anthocyanin levels, and antioxidant capacity in the peel and pulp of 16 mixed-species rabbiteye hybrids ( Vaccinium ashei Readexa0×xa0 Vaccinium spp.), one rabbiteye cultivar ( V. ashei ) and two highbush hybrids ( Vaccinium corymbosum ). Peel tissue had, on average, about four times higher levels of α-glucosidase inhibitory activity than pulp, and exhibited significantly higher levels of all other measured activities, even though the peel comprised only a small portion of the fruit. Significant variations in the levels of antioxidant activity were observed; however, no consistent differences were observed between the hybrids with various species composition. Significant positive correlations ( r ≥xa00.84) were found among α-glucosidase inhibitory activity, total anthocyanin (TA) and phenolic levels, and scavenging activity against ROO∙, ∙OH, 1 O 2 and H 2 O 2 radicals in the extracts from the peel and pulp. There was a high correlation observed between α-glucosidase inhibitory activity levels and ROO∙(ORAC) peel ( r =xa00.95). A similarly high correlation with TA peel ( r =xa00.93) suggests that TA would be a suitable assay proxy if a broader genotypic evaluation of blueberry genotypes is desired.

Recovery of Phytophthora ramorum in plant tissue with mixed infections

This study was performed to investigate the frequency with which P. ramorum would be isolated from host tissue co-infected with P. ramorum as well as an indigenous Phytophthora species or P. kernoviae. Three separate experiments were tested in a similar manner using different combinations of pathogens and hosts. Overall for any of the individual experiments, very few segments did not have any growth of Phytophthora spp. For mixed infections of P. ramorum and P. kernoviae, a difference was observed between isolating both of the species and P. ramorum only on rhododendron. The data showed that P. ramorum or P. kernoviae will not be detected 29 or 12% of the time, respectively, in infected Rhododendron sp. Phytophthora kernoviae was not detected alone in mixed infections with P. ramorum on Pieris japonica. When two different P. ramorum isolates were co-inoculated individually with one P. citricola isolate, there was a significant difference between isolating P. ramorum and isolating both species. These results confirm that choice of host species used for baiting can strongly influence detection results. For example, if P. japonica were used for baiting in mixed infections, there is a 55% chance that P. kernoviae would not be detected. This study highlights the difficulty in being confident in isolating and identifying an individual Phytophthora sp. from host material when mixed infections are present, and emphasizes the importance of a large sample size in order to increase the chances to recover all possible different species in a mixed infection.