Integrated Effects of Genetic Resistance and Prothioconazole + Tebuconazole Application Timing on Fusarium Head Blight in Wheat.

Integrated Fusarium head blight (FHB) management programs consisting of different combinations of cultivar resistance class and an application of the fungicide prothioconazole + tebuconazole at or after 50% early anthesis were evaluated for efficacy against FHB incidence (INC; percentage of diseased spikes), index (IND; percentage of diseased spikelets per spike), Fusarium damaged kernel (FDK), deoxynivalenol (DON) toxin contamination, grain yield, and test weight (TW) in inoculated field trials conducted in 11 U.S. states in 2014 and 2015. Mean log response ratios and corresponding percent control values for INC, IND, FDK, and DON, and mean differences in yield and TW relative to a nontreated, inoculated susceptible check (S_CK), were estimated through network meta-analyses as measures of efficacy. Results from the analyses were then used to estimate the economic benefit of each management program for a range of grain prices and fungicide applications costs. Management programs consisting of a moderately resistant (MR) cultivar treated with the fungicide were the most efficacious, reducing INC by 60 to 69%, IND by 71 to 76%, FDK by 66 to 72%, and DON by 60 to 64% relative to S_CK, compared with 56 to 62% for INC, 68 to 72% for IND, 66 to 68% for FDK, and 58 to 61% for DON for programs with a moderately susceptible (MS) cultivar. The least efficacious programs were those with a fungicide application to a susceptible (S) cultivar, with less than a 45% reduction of INC, IND, FDK, or DON. All programs were more efficacious under conditions favorable for FHB compared with less favorable conditions, with applications made at 50% early anthesis being of comparable efficacy to those made 2 to 7 days later. Programs with an MS cultivar resulted in the highest mean yield increases relative to S_CK (541 to 753 kg/ha), followed by programs with an S cultivar (386 to 498 kg/ha) and programs with an MR cultivar (250 to 337 kg/ha). Integrated management programs with an MS or MR cultivar treated with the fungicide at or after 50% early anthesis were the most likely to result in a 50 or 75% control of IND, FDK, or DON in a future trial. At a fixed fungicide application cost, these programs were $4 to $319/MT more economically beneficial than corresponding fungicide-only programs, depending on the cultivar and grain price. These findings demonstrate the benefits of combining genetic resistance with a prothioconazole + tebuconazole treatment to manage FHB, even if that treatment is applied a few days after 50% early anthesis.

First Report of Charcoal Rot Caused by Macrophomina phaseolina in Soybean in New York.

Charcoal rot of soybean (Glycine max (L.) Merr.), incited by Macrophomina phaseolina (Tassi) Goidanich, is commonly found in much of the southern soybean production region of the United States, where it can be a major contributor to yield loss in warm, dry seasons (4). The disease has also been reported in northern states including Michigan, Minnesota, North Dakota, and Wisconsin (1,2,3). In early July of the warmer and drier than normal 2012 growing season, wilted soybean plants were observed and collected for diagnosis from a field in Cayuga County, NY. The diseased plants showed brown discoloration of the stem and roots but the diagnostic microsclerotia of M. phaseolina had not yet developed in the epidermis. Cut pieces of roots and crowns were surface-sterilized in 20% sodium hypochlorite, rinsed in sterile distilled water, and incubated on potato dextrose agar at room temperature with a 12-h photoperiod for 7 days. The pathogen was observed growing out of many of the crown and root pieces, and produced abundant microsclerotia as described by Smith and Wyllie (4). Pathogenicity of isolate Mp001NY12 was confirmed using a cut stem inoculation method. Five soybean plants were grown to the unifoliate stage and inoculated by cutting the stem above the unifoliate leaves and applying a fully colonized agar plug to the wound. Within 2 weeks, infection was obvious with microsclerotia formed in the epidermis of all infected plants, and M. phaseolina was reisolated from all infected plants. DNA of isolate Mp001NY12 was extracted from colonies grown on PDA with PrepMan Ultra DNA extraction kit (Applied Biosystems, Foster City, CA), and the DNA was submitted for ITS sequencing with the ITS1 and ITS4 primers used for PCR amplification. The ITS sequence (GenBank Accession No. KC800709) of Mp001NY12 was compared to those in the NCBI GenBank database using a BLAST search, and had 99% nucleotide sequence identity with M. phaseolina (accessions JX945170, FJ415067, and EU250575). To the best of our knowledge, charcoal rot has not been reported previously on soybean or other plant hosts in New York or in other states in the northeastern United States. References: (1) Baird et al. Mycopathologia 170:169, 2010. (2) C. A. Bradley and L. E. del Rio. Plant Dis. 87:601, 2003. (3) M. E. ElAraby et al. Plant Dis. 87:202, 2003. (4) G. S. Smith and T. D. Wyllie. Charcoal rot. Page 29 in: Compendium of Soybean Diseases, 4th ed. G. L. Hartman et al., eds. APS Press, St. Paul, MN, 1999.

Mother counts: how effects of environmental contaminants on maternal care could affect the offspring and future generations.

Various compounds of anthropogenic origin represent environmental contaminants (EC) that penetrate the food chain and are frequently detected in human milk and maternal blood at the time of delivery. These ECs can affect the development of the fetus and can be transferred to the newborn during lactation. Many studies have used animal models to study the impact of ECs on the development of the nervous system and have reported effects of early exposure on neural and neuroendocrine systems and on behavior, when the exposed animals are tested as adults. Some of these effects persist across generations and may involve epigenetic mechanisms. The majority of these studies in developmental toxicology treat the pregnant or lactating animal with ECs in order to deliver the contaminants to the developing offspring. Almost universally, the mother is viewed as a passive conduit for the ECs, and maternal behavior is rarely assessed. Here we review the literature on the effects of ECs on maternal care and find mounting evidence that important components of the care given to the offspring are affected by maternal exposure to different ECs. Some of these changes in maternal behavior appear to be secondary to changes in the behavior and/or stimulus properties of the exposed offspring, but others are likely to be direct effects of the ECs on the maternal nervous and endocrine systems. Considering the extent to which the quality of maternal care affects the development of the offspring, it becomes imperative to determine the contributions that changes in maternal behavior make to the deficits traditionally ascribed solely to direct effects of ECs on the developing organism. Given the complexity and importance of mother-infant interactions, future research on developmental toxicology must consider the effects of ECs not only on the offspring, but also on the mother and on the interactions and social bond between mother and infant.

Exposure to PCB 77 affects partner preference but not sexual behavior in the female rat.

In rats, exposure to the polychlorinated biphenyl congener 3, 4, 3', 4'-tetrachlorobiphenyl (PCB 77) affects the brain and behavior of the offspring as well as the maternal behavior of the dams. In the present study, a cross-fostering design was used to examine the effects of pre- and/or postnatal exposure to PCB 77 on sexual behavior and partner preference in female rats, and to determine the role of altered maternal behavior in the mediation of these effects. Pregnant rats were treated with oil or PCB dissolved in oil (2 mg/kg b.w.) on gestation days 6-18 and then given pups that had been exposed to either the oil vehicle or PCB during gestation. As adults, the female offspring were tested for partner preference (that is, whether they preferred to spend time with a sexually receptive female or a sexually active male) and sexual behavior. None of the treatments affected female sexual behavior. However, both double exposure and postnatal exposure diminished the animals' preference for a male over a female stimulus, but partner preference was not affected by prenatal exposure alone. There were no significant correlations between the changes in partner preferences due to PCB exposure and the amount of maternal grooming and licking received by the treated litters. Thus, female partner preference is affected by early PCB exposure, and the effects depend upon whether the exposure is in utero or via lactation and may be independent of any effects of the PCB on maternal care.

A cross-fostering analysis of the effects of PCB 77 on the maternal behavior of rats.

Polychlorinated biphenyls (PCBs) are environmental contaminants known to cause multiple behavioral and developmental problems in humans and animals. In rats, gestational exposure to the PCB congener 3, 4, 3', 4'-tetrachlorobiphenyl (PCB 77) affects the brain and behavior of the offspring as well as the maternal behavior of the dams. Whether the behavior of dams is affected by direct effects of the contaminant or indirectly by actions of the PCB on the developing offspring is not known. We investigated this question using a cross-fostering paradigm in which pregnant rats were exposed to either oil vehicle or 2 mg/kg of PCB 77 on gestational days 6 through 18, and then raised pups that had been exposed to either oil or PCB 77 during gestation. Maternal behavior was observed on postnatal days 1, 2, 4 and 6. Some of the effects on maternal behavior, including an increase in the frequency of nursing bouts and in the amount of maternal autogrooming, can be ascribed to prenatal exposure of the litters to the PCB. Other behavioral effects, including an increase in time on the nest and in the amount of pup grooming as well as a reduction in high-crouch nursing, appear to be due to both direct effects of the PCB on the dams and effects mediated by changes in the offspring. Our results show that exposure to PCB 77 can have complex effects on the behavioral interactions between the dams and their litters with a potential impact on the development of the offspring.

Exposure to PCB 77 affects the maternal behavior of rats.

Polychlorinated biphenyls are environmental contaminants known to affect neurobehavioral development in many laboratory studies using different animal models. Because of their bioaccumulation and long half-life they are a serious concern for our own species. The dioxin-like PCB congener 3,4,3',4'-tetrachlorobiphenyl (PCB 77) has estrogenic and anti-estrogenic properties, and has been shown to affect brain chemistry and behavior of developing rats when administered during gestation. Since many developmental outcomes in mammals depend upon the type of maternal care provided by the dams, we investigated the effects of two doses of PCB 77 (2 and 4 mg/kg administered during gestational days 6-18) on the maternal behavior of the treated dams. Both doses of PCB 77 reduced the amount of nursing time in which the dams displayed the high-crouch posture over postnatal days 1-6. In addition, the high dose increased the amount of maternal licking and grooming of the litters and the amount of time the dams spent on the nest. The high dose also increased pup mortality, and both doses reduced the weight gain of the litters during the first 6 days of life. These results document effects of PCB 77 on maternal behavior and serve to raise questions about the importance of maternal contributions to the developmental effects of this and similar contaminants.

Sulfate content and specific glycosaminoglycan backbone of perlecan are critical for perlecan's enhancement of islet amyloid polypeptide (amylin) fibril formation.

Islet amyloidosis is characterized by the deposition and accumulation of amylin in pancreatic beta-cells and is observed in 90% of patients with type 2 diabetes. Previous studies have also revealed the presence of the specific heparan sulfate proteoglycan, perlecan, colocalized to islet amyloid deposits, similar to perlecan's known involvement with other amyloid proteins. In the present study, perlecan purified from the Engelbreth-Holm-Swarm (EHS) tumor was used to define perlecan's interactions with amylin (i.e., islet amyloid polypeptide) and its effects on amylin fibril formation. Using a solid phase-binding immunoassay, human amylin, but not rat amylin, bound immobilized EHS perlecan with a single dissociation constant (Kd) = 2.75 x 10(-6) mol/l. The binding of human amylin to perlecan was similarly observed using perlecan heparan sulfate glycosaminoglycans (GAGs), and was completely abolished by 10 micromol/l heparin. Using thioflavin T fluorometry, Congo red staining, and electron microscopy methodology, intact perlecan was found to enhance amylin fibril formation in a dosage-dependent manner, with the majority of these effects attributed to the heparan sulfate GAG chains of perlecan. Other sulfated GAGs and related macromolecules were also effective in the enhancement of amylin fibril formation in the order of heparin > heparan sulfate > chondroitin-4-sulfate = dermatan sulfate = dextran sulfate > pentosan polysulfate, implicating the importance of the specific GAG/carbohydrate backbone. The sulfate content of heparin/heparan sulfate was also important for the enhancement of amylin fibril formation in the order of heparin > N-desulfated N-acetylated heparin > completely desulfated N-sulfated heparin > completely desulfated N-acetylated heparin. These studies suggest that the enhancement effects of perlecan on amylin fibril formation are mediated primarily by both specific GAG chain backbone and GAG sulfate content, and implicate perlecan as an important macromolecule that is likely involved in the pathogenesis of islet amyloidosis.

Novel purification and detailed characterization of perlecan isolated from the Engelbreth-Holm-Swarm tumor for use in an animal model of fibrillar A beta amyloid persistence in brain.

Co-infusion of the specific heparan sulfate proteoglycan (HSPG), perlecan, and beta-amyloid protein (A beta) into rodent hippocampus leads to a consistent animal model to study the effects of fibrillar A beta amyloid in brain [Snow, A.D. et al. (1994) Neuron 12, 219-234]. In the present study, we describe our rapid novel method of perlecan isolation. The isolation method does not require cesium chloride centrifugation and exploits a newly discovered aggregating property of a approximately 220 kDa PG observed during gel filtration chromatography, which allowed it to be affectively separated from non-aggregating perlecan. Fifty or 100 g of EHS tumor were routinely extracted using 4 M guanidine-HCl, followed by anion-exchange and gel filtration chromatography. SDS-PAGE (before and after digestion with heparitinase/heparinase or nitrous acid) followed by staining with silver demonstrated no other contaminating proteins in the perlecan preparations. Western blots using a specific perlecan core protein antibody (HK-102) following heparitinase digestion showed a characteristic doublet at 400 and 360 kDa indicative of intact perlecan core protein. Absence of contamination by other basement membrane components produced by the EHS tumor was confirmed by absence of immunoreactive bands on Western blots using antibodies against laminin, fibronectin, or type IV collagen. One week continuous co-infusion of perlecan obtained from this methodology, with A beta (1-40) into rodent hippocampus, led to deposition of fibrillar A beta amyloid in 100% (10 of 10) of animals. The detailed protocol for isolation and characterization of perlecan from EHS tumor ensures perlecan of the highest quality, and maximizes the potential effects of A beta amyloid deposition/persistence in brain using the animal model. High quality perlecan obtained from this novel isolation method will also allow future studies utilizing in vitro assays to determine the potential interactions of this specific HSPG with other macromolecules.

Ca2+ signaling requirements for long-term depression in the hippocampus.

It has been hypothesized that the direction of synaptic weight change elicited by synaptic activity depends on the magnitude of the activity-dependent rise in intracellular Ca2+ concentration. Several aspects of this hypothesis were examined at the Schaffer collateral CA1 synapse, where both long-term depression (LTD) and long-term potentiation (LTP) can be elicited and are Ca2+ dependent. Brief tetanic stimulation, which normally generated LTP, could induce LTD when Ca2+ entry via NMDA receptors was limited either by moderate concentrations of D-APV or by voltage clamping cells at negative membrane potentials. Repetitive activation of voltage-dependent Ca2+ channels in the absence of afferent stimulation could also elicit an LTD that was Ca2+ dependent and was occluded by prior generation of homosynaptic LTD using prolonged low evidence that the minimal requirements for inducing LTD involve simply a transient influx of Ca2+ into the postsynaptic cell, via either NMDA receptors or voltage-dependent Ca2+ channels.

Long-term survival followed by degradation of neurofilament proteins in severed mauthner axons of goldfish.

The morphology and protein composition of intact and severed Mauthner axons (M-axons) from goldfish were examined on electron micrographs, sodium dodecyl sulfate gels, and immunoblots. Neurofilaments were the most common cytoskeletal element on electron micrographs, and neurofilament proteins (NFPs) were the most intensely silver-stained bands in M-axoplasm microdissected from control M-axons. NFPs at about 235, 145, 123, 105, 80, and 60 kD in M-axoplasm were identified with four monoclonal and three polyclonal antibodies. Similar immunoblots of samples of the M-axon myelin sheath (M-sheath) showed no reactivity to antibodies against NFPs. For up to 62 days following spinal cord severance in goldfish maintained at 15 degrees C, the ultrastructure, protein banding pattern, and anti-NFP immunoreactivity of several distal segments of M-axons did not change compared with control M-axons. At 62 to 81 days after severance, novel bands appeared in many silver-stained gels and anti-NFP immunoblots of distal M-axons. NFP bands completely disappeared from distal M-axon segments of some M-axons as early as 72 days after severance. However, NFP bands persisted in some distal segments for up to 81 days after severance. The degradation of NFPs occurred equally along the entire length of a distal M-axon segment, that is, there was no indication of a proximal-to-distal or distal-to-proximal sequence of NFP degradation in distal segments of severed M-axons. These biochemical data were consistent with morphological data that showed little change in the diameter or ultrastructure of severed M-axons held at 15 degrees C for about 2 months followed by a rapid collapse of the entire distal segment at 72 to 85 days postseverance.

Induction in the rat hippocampus of long-term potentiation (LTP) and long-term depression (LTD) in the presence of a nitric oxide synthase inhibitor.

Several recent studies have suggested a critical role for nitric oxide (NO) production in hippocampal LTP and LTD. In this study we show that normal LTP and LTD can be induced in rat hippocampal slices incubated in the NO synthase inhibitor L-NG-nitroarginine (NOArg) (100 microM). A test of NMDA-stimulated cGMP production demonstrated that incubation of slices in 100 microM NOArg effectively inhibited NO synthase. Our results suggest that NO synthase activity may not be required for the generation of LTP or LTD in CA1 of rat hippocampus.

Coccidioidomycosis in a California sea lion (Zalophus californianus).

Coccidioidomycosis in an adult male California sea lion (Zalophus californianus) is described. The animal was housed in a zoo in Tucson, Arizona, for approximately 5 years. This is believed to be the first reported case of coccidioidomycosis in a marine mammal.