Search

‘Candidatus Liberibacter asiaticus’ (Las) is the prominent species of Liberibacter associated with huanglongbing, a devastating disease of citrus worldwide. In this study, we report the identification of an ∼8.3-kb DNA region of the Las genome containing eight putative open reading frames flanked by two inverted repeats, which was not present in the Las str. psy62 genome. Comparisons with other genome sequences established this region as a unique genetic element associated with genome plasticity/instability. Primers specific for both the presence (Las wild type) and absence (Las mutant) of this region were designed to study the population dynamics and host adaptation of the two strains. Las populations with and/or without the wild-type strain were detected and differentiated in >2,300 samples that included psyllids, periwinkle, and several species of citrus. In psyllids, although a mixed population of the wild type and mutant was observed in most samples (88%), the wild-type Las was detected alone at a rate of 11%. In contrast, none of the infected citrus plants were positive for the wild type alone, which harbored either the mutant strain alone (8%) or a mixed population of the mutant and wild type (92%). Furthermore, the dynamics of these two major Las populations varied with different citrus hosts, whereas an in-depth study on grapefruit that did not rapidly succumb to disease revealed that the population of mutant alone increased with time, indicating that the absence of this genetic element is associated with the fitness of Las in planta under the selection pressure of its host.

Understanding vector-host interactions is crucial for evaluating the role of mosquito species in enzootic cycling and epidemic/epizootic transmission of arboviruses, as well as assessing vertebrate...

The hybrid cattail Typha xglauca Godr. is morphologically intermediate between Typha latifolia L. and Typha angustifolia L. We propose a method to distinguish these taxa based on morphology throughout emergent life stages. We evaluated four traits of gross morphology and four more from microscopic examination of leaf cross sections. All eight traits were assessed for 45 cattails collected from various types of wetlands in Manitoba and Saskatchewan, Canada, these shoots being identified by sequencing of microsatellite DNA as 17 specimens of T. latifolia and 14 specimens each for T. angustifolia and T. xglauca. Decision-tree analysis indicated that the single trait of mean leaf-apex angle for all leaves on the shoot was able to correctly align all specimens with their genetic identities. However, single-leaf measurements for leaf-apex angle were successful in this regard in only 80–85% of attempts. Ignoring mean leaf-apex angle and using a combination of all remaining seven traits, 44-out-of-45 samples were aligned correctly. Measurement of mean leaf-apex angle for a shoot can be used to distinguish reliably the parental Typha species and the hybrid. The remaining seven morphological traits can then be employed to provide additional support for determinations, possibly as an alternative to genetic determinations.

The detrimental effects of Huanglongbing (HLB) on citrus are well known and so is the need for effective methods to combat this disease. Solar thermotherapy (ST), one of the management methods to help alleviate some of the negative effects of HLB, is the process of heating trees to therapeutic temperatures by encompassing the tree within a plastic structure (Visqueen) to harness the sun's natural energy. ST was applied to mature ‘Valencia’ citrus trees in three locations. Tree vigor, yield, and Candidatus Liberibacter asiaticus (Las) titer in leaves were monitored for two consecutive seasons post ST treatment, while fruit and juice quality were evaluated at the end of the second season. ST promoted an increase in canopy density in most groves tested and did not have a significant effect on fruit quantity, despite the prolonged exposure of the trees to increased temperatures. Moreover, Las titer was reduced in both leaves and juice processed from affected trees post ST. With respect to fruit quality, the ratio of total soluble solids to titratable acidity was higher post ST compared to controls as was the sucrose content of the juice in most treated groves, whereas limonin was higher in juice from control trees. Principal components analysis of aroma volatiles showed significant differences between juice from treated versus control trees with volatiles imparting top-note and freshness to orange juice, such as acetaldehyde, hexanal, Z-3-hexenol and linalool, being higher in juice from treated trees. Taste panels confirmed that flavor differences existed, ultimately showing that ST resulted in improved juice flavor in well-managed groves. Although the effects of ST on the trees appear temporary, such outcomes emphasize its utility when used as part of an integrated management strategy for HLB-affected citrus, imparting beneficial horticultural responses with minimal to positive effects on subsequent juice flavor.

Targeting androgen signaling with the second-generation anti-androgen drugs, such as enzalutamide (Enza), abiraterone (Abi), apalutamide (Apal), and darolutamide (Daro), is the mainstay for the treatment of castration-resistant prostate cancer (CRPC). While these treatments are effective initially, resistance occurs frequently. Continued expression of androgen receptor (AR) and its variants such as AR-V7 despite AR-targeted therapy contributes to treatment resistance and cancer progression in advanced CRPC patients. This highlights the need for new strategies blocking continued AR signaling. Here, we identify a novel AR/AR-V7 degrader (ARVib) and found that ARVib effectively degrades AR/AR-V7 protein and attenuates AR/AR-V7 downstream target gene expression in prostate cancer cells. Mechanistically, ARVib degrades AR/AR-V7 protein through the ubiquitin-proteasome pathway mediated by HSP70/STUB1 machinery modulation. ARVib suppresses HSP70 expression and promotes STUB1 nuclear translocation, where STUB1 binds to AR/AR-V7 and promotes its ubiquitination and degradation. ARVib significantly inhibits resistant prostate tumor growth and improves enzalutamide treatment in vitro and in vivo. These data suggest that ARVib has potential for development as an AR/AR-V7 degrader to treat resistant CRPC.

Purpose: Fumarate hydratase–deficient renal cell carcinoma (FH-deficient RCC) is a rare but lethal subtype of RCC. Little is known about the genomic profile of FH-deficient RCC, and the therapeutic options for advanced disease are limited. To this end, we performed a comprehensive genomics study to characterize the genomic and epigenomic features of FH-deficient RCC. Experimental Design: Integrated genomic, epigenomic, and molecular analyses were performed on 25 untreated primary FH-deficient RCCs. Complete clinicopathologic and follow-up data of these patients were recorded. Results: We identified that FH-deficient RCC manifested low somatic mutation burden (median 0.58 mutations per megabase), but with frequent somatic copy-number alterations. The majority of FH-deficient RCCs were characterized by a CpG sites island methylator phenotype, displaying concerted hypermethylation at numerous CpG sites in genes of transcription factors, tumor suppressors, and tumor hallmark pathways. However, a few cases (20%) with low metastatic potential showed relatively low DNA methylation levels, indicating the heterogeneity of methylation pattern in FH-deficient RCC. Moreover, FH-deficient RCC is potentially highly immunogenic, characterized by increased tumor T-cell infiltration but high expression of immune checkpoint molecules in tumors. Clinical data further demonstrated that patients receiving immune checkpoint blockade–based treatment achieved improved progression-free survival over those treated with antiangiogenic monotherapy (median, 13.3 vs. 5.1 months; P = 0.03). Conclusions: These results reveal the genomic features and provide new insight into potential therapeutic strategies for FH-deficient RCC.

N-methyl-D-aspartate receptors (NMDARs) are excitatory ionotropic glutamate receptors expressed throughout the CNS, including in the spinal dorsal horn. The GluN2 subtypes of NMDAR subunit, which include GluN2A, GluN2B, and GluN2D in the dorsal horn, confer NMDARs with structural and functional variability, enabling heterogeneity in synaptic transmission and plasticity. Despite essential roles for NMDARs in physiological and pathological pain processing, the distribution and function of these specific GluN2 isoforms across dorsal horn laminae remain poorly understood. Surprisingly, there is a complete lack of knowledge of GluN2 expression in female rodents. We, therefore, investigated the relative expression of specific GluN2 variants in the dorsal horn of lumbar (L4/L5) spinal cord from both male and female rats. In order to detect synaptic GluN2 isoforms, we used pepsin antigen-retrieval to unmask these highly cross-linked protein complexes. We found that GluN2B and GluN2D are preferentially localized to the pain-processing superficial regions of the dorsal horn in males, while only GluN2B is predominantly localized to the superficial dorsal horn of female rats. The GluN2A subunit is diffusely localized to neuropil throughout the dorsal horn of both males and females, while GluN2B and GluN2D immunolabelling are found both in the neuropil and on the soma of dorsal horn neurons. Finally, we identified an unexpected enhanced expression of GluN2B in the medial division of the superficial dorsal horn, but in males only. These sex-specific localization patterns of GluN2-NMDAR subunits across dorsal horn laminae have significant implications for the understanding of divergent spinal mechanisms of pain processing.

Pesticide resistance in arthropod vectors of disease agents is a growing issue globally. Despite the importance of resistance monitoring to inform mosquito control programs, no regional monitoring programs exist in the United States. The Northeastern Regional Center for Excellence in Vector-Borne Diseases (NEVBD) is a consortium of researchers and public health practitioners with a primary goal of supporting regional vector control activities. NEVBD initiated a pesticide resistance monitoring program to detect resistant mosquito populations throughout the northeastern United States. A regionwide survey was distributed to vector control agencies to determine needs and refine program development and in response, a specimen submission system was established, allowing agencies to submit Culex pipiens (L.) (Diptera:Culicidae) and Aedes albopictus (Skuse) (Diptera: Culicidae) for pesticide resistance testing. NEVBD also established larvicide resistance diagnostics for Bacillus thuringiensis israelensis (Bti) and methoprene. Additional diagnostics were developed for Cx. pipiens resistance to Lysinibacillus sphaericus. We received 58 survey responses, representing at least one agency from each of the 13 northeastern U.S. states. Results indicated that larvicides were deployed more frequently than adulticides, but rarely paired with resistance monitoring. Over 18,000 mosquitoes were tested from six states. Widespread low-level (1 × LC-99) methoprene resistance was detected in Cx. pipiens, but not in Ae. albopictus. No resistance to Bti or L. sphaericus was detected. Resistance to pyrethroids was detected in many locations for both species. Our results highlight the need for increased pesticide resistance testing in the United States and we provide guidance for building a centralized pesticide resistance testing program.

Docetaxel and cabazitaxel-based taxane chemotherapy are critical components in the management of advanced prostate cancer. However, their efficacy is hindered due to de novo presentation with or the development of resistance. Characterizing models of taxane-resistant prostate cancer will lead to creation of strategies to overcome insensitivity. We have previously characterized docetaxel-resistant C4-2B and DU145 cell line derivatives, TaxR and DU145-DTXR, respectively. In the present study, we characterize cabazitaxel-resistant derivative cell lines created from chronic cabazitaxel exposure of TaxR and DU145-DTXR cells, CabR and CTXR, respectively. We show that CabR and CTXR cells are robustly resistant to both taxanes but retain sensitivity to antiandrogens. Both CabR and CTXR cells possess increased expression of ABCB1, which is shown to mediate resistance to treatment. Interestingly, we also present evidence for coordinated overexpression of additional genes present within the 7q21.12 gene locus where ABCB1 resides. This locus, known as the ABCB1 amplicon, has been demonstrated to be amplified in multidrug-resistant tumor cells, but little is known regarding its role in prostate cancer. We show that two ABCB1-amplicon genes other than ABCB1, RUNDC3B and DBF4, promote cellular viability and treatment resistance in taxane-resistant prostate cancer models. We present evidence that coordinated amplification of ABCB1-amplicon genes is common in a subset of prostate cancer patients. These data together suggest that ABCB1-amplicon activation plays a critical role in taxane resistance.

TFE3-translocation renal cell carcinoma (TFE3-tRCC) is a rare and heterogeneous subtype of kidney cancer with no standard treatment for advanced disease. We describe comprehensive molecular characteristics of 63 untreated primary TFE3-tRCCs based on whole-exome and RNA sequencing. TFE3-tRCC is highly heterogeneous, both clinicopathologically and genotypically. ASPSCR1-TFE3 fusion and several somatic copy number alterations, including the loss of 22q, are associated with aggressive features and poor outcomes. Apart from tumors with MED15-TFE3 fusion, most TFE3-tRCCs exhibit low PD-L1 expression and low T-cell infiltration. Unsupervised transcriptomic analysis reveals five molecular clusters with distinct angiogenesis, stroma, proliferation and KRAS down signatures, which show association with fusion patterns and prognosis. In line with the aggressive nature, the high angiogenesis/stroma/proliferation cluster exclusively consists of tumors with ASPSCR1-TFE3 fusion. Here, we describe the genomic and transcriptomic features of TFE3-tRCC and provide insights into precision medicine for this disease., TFE3-translocation renal cell carcinoma (TFE3-tRCC) is a rare subtype of kidney cancer with no standard treatment options for the advanced disease. Here, the authors perform genomic and transcriptomic profiling of 63 untreated primary TFE3-tRCC tumours and reveal potential therapeutic targets.

INTRODUCTION AND OBJECTIVE:Inhibition of poly (ADP-ribose) polymerase (PARP) is an exciting treatment strategy recently approved for prostate cancer patients with homologous recombination repair de...

Aedes albopictus is a vector of arboviruses with high rates of morbidity and mortality. The northern limit of Ae. albopictus in the northeastern United States runs through New York state (NYS) and Connecticut. We present a landscape-level analysis of mosquito abundance measured by daily counts of Ae. albopictus from 338 trap sites in 12 counties during May-September 2017. During the study period, the mean number of Ae. albopictus caught per day of trapping across all sites was 3.21. We constructed four sets of negative binomial generalized linear models to evaluate how trapping methodology, land cover, as well as temperature and precipitation at multiple time intervals influenced Ae. albopictus abundance. Biogents-Sentinel (BGS) traps were 2.78 times as efficient as gravid traps and 1.49 times as efficient as CO2-baited CDC light traps. Greater proportions of low- and medium-intensity development and low proportions of deciduous cover around the trap site were positively associated with increased abundance, as were minimum winter temperature and March precipitation. The cumulative precipitation within a 28-day time window before the date of collection had a nonlinear relationship with abundance, such that greater cumulative precipitation was associated with increased abundance until approximately 70 mm, above which there was a decrease in abundance. We concluded that populations are established in Nassau, Suffolk, and New York City counties in NYS; north of these counties, the species is undergoing population invasion and establishment. We recommend that mosquito surveillance programs monitoring the northward invasion of Ae. albopictus place BGS traps at sites chosen with respect to land cover. Centers for Disease Control and PreventionUnited States Department of Health & Human ServicesCenters for Disease Control & Prevention - USA [U01CK000509-01] This publication was supported by the Cooperative Agreement Number U01CK000509-01, funded by the Centers for Disease Control and Prevention.

Contains fulltext : 219496.pdf (Publisher’s version ) (Closed access) DNA methylation is the most studied modification in human population epigenetics. Its information content can be explored in 2 principal ways - epigenome-wide association studies and epigenetic age. The latter likely reflects cellular/biological age and works with impressive accuracy across most tissues. In adults, it associates with various environments and health. However, current epigenetic clocks are not very accurate in the pediatric age range perhaps because DNA methylation changes much faster in children. Addressing this crucial gap, we created a precise tool to estimate DNA methylation age specific to pediatric buccal epithelial cells. This tool has the potential to become the standard reference for epigenetic studies broadly relevant to child development across the spectrum from health to disease.The development of biological markers of aging has primarily focused on adult samples. Epigenetic clocks are a promising tool for measuring biological age that show impressive accuracy across most tissues and age ranges. In adults, deviations from the DNA methylation (DNAm) age prediction are correlated with several age-related phenotypes, such as mortality and frailty. In children, however, fewer such associations have been made, possibly because DNAm changes are more dynamic in pediatric populations as compared to adults. To address this gap, we aimed to develop a highly accurate, noninvasive, biological measure of age specific to pediatric samples using buccal epithelial cell DNAm. We gathered 1,721 genome-wide DNAm profiles from 11 different cohorts of typically developing individuals aged 0 to 20 y old. Elastic net penalized regression was used to select 94 CpG sites from a training dataset (n = 1,032), with performance assessed in a separate test dataset (n = 689). DNAm at these 94 CpG sites was highly predictive of age in the test cohort (median absolute error = 0.35 y). The Pediatric-Buccal-Epigenetic (PedBE) clock was characterized in additional cohorts, showcasing the accuracy in longitudinal data, the performance in nonbuccal tissues and adult age ranges, and the association with obstetric outcomes. The PedBE tool for measuring biological age in children might help in understanding the environmental and contextual factors that shape the DNA methylome during child development, and how it, in turn, might relate to child health and disease. 7 p.

The recent Zika virus (ZIKV) and chikungunya virus epidemics highlight the explosive nature of arthropod-borne viruses (arboviruses) transmitted by Aedes spp. mosquitoes1,2. Vector competence and the extrinsic incubation period (EIP) are two key entomological parameters used to assess the public health risk posed by arboviruses3. These are typically measured empirically by offering mosquitoes an infectious blood meal and temporally sampling mosquitoes to determine the infection and transmission status. This approach has been used for the better part of a century; however, it does not accurately capture the biology and behaviour of many mosquito vectors that refeed frequently (every 2–3 d)4. Here, we demonstrate that acquisition of a second non-infectious blood meal significantly shortens the EIP of ZIKV-infected Aedes aegypti by enhancing virus dissemination from the mosquito midgut. Similarly, a second blood meal increases the competence of this species for dengue virus and chikungunya virus as well as Aedes albopictus for ZIKV, suggesting that this phenomenon may be common among other virus–vector pairings and that A. albopictus might be a more important vector than once thought. Blood-meal-induced microperforations in the virus-impenetrable basal lamina that surrounds the midgut provide a mechanism for enhanced virus escape. Modelling of these findings reveals that a shortened EIP would result in a significant increase in the basic reproductive number, R0, estimated from experimental data. This helps to explain how A. aegypti can sustain explosive epidemics such as ZIKV despite relatively poor vector competence in single-feed laboratory trials. Together, these data demonstrate a direct and unrecognized link between mosquito feeding behaviour, EIP and vector competence. This study reports that flavivirus transmission by mosquitoes to mice is increased if mosquitoes feed subsequently on non-infectious blood, possibly because feeding causes microperforations in the gut. Modelling shows this could explain how A. aegypti can sustain an explosive epidemic such as Zika virus despite its perceived poor vector competence.

West Nile virus (WNV) is transmitted by mosquitoes and can cause severe disease, including meningoencephalitis. AgBR1 is a mosquito salivary protein that enhances Aedes aegypti mosquito-borne Zika virus pathogenesis in mice. Here, we show that AgBR1 antibodies reduce the initial West Nile viral load and delay lethal infection after feeding by an infected Aedes aegypti mosquito. Targeting AgBR1 may therefore be incorporated into strategies to prevent mosquito-transmitted West Nile virus infection.

The bilaterally symmetrical, feeding larval stage is an ancestral condition in echinoderms. However, many echinoderms have evolved abbreviated development and form a pentamerous juvenile without a feeding larva. Abbreviated development with a non-feeding vitellaria larva is found in five families of brittle stars, but very little is known about this type of development. In this study, the external anatomy, ciliary bands, neurons, and muscles were examined in the development of the brooded vitellaria larva of Ophioplocus esmarki. The external morphology throughout development shows typical vitellaria features, including morphogenetic movements to set up the vitellaria body plan, an anterior preoral lobe, a posterior lobe, transverse ciliary bands, and development of juvenile structures on the mid-ventral side. An early population of neurons forms at the base of the preoral lobe at the pre-vitellaria stage after the initial formation of the coelomic cavities. These early neurons may be homologous to the apical neurons that develop in echinoderms with feeding larval forms. Neurons form close to the ciliary bands, but the vitellaria larva lacks the tracts of neurons associated with the ciliary bands found in echinoderms with feeding larvae. Additional neurons form in association with the axial complex and persist into the juvenile stage. Juvenile nerves and muscles form with pentamerous symmetry in the late vitellaria stage in a manner similar to their development within the late ophiopluteus larva. Even though O. esmarki is a brooding brittle star, its developmental sequence retains the general vitellaria shape and structure; however, the vitellaria larvae are unable to swim in the water column.

Cognitive aging varies tremendously across individuals and is often accompanied by regionally specific reductions in gray matter (GM) volume, even in the absence of disease. Rhesus monkeys provide a primate model unconfounded by advanced neurodegenerative disease, and the current study used a recognition memory test (delayed non-matching to sample; DNMS) in conjunction with structural imaging and voxel-based morphometry (VBM) to characterize age-related differences in GM volume and brain-behavior relationships. Consistent with expectations from a long history of neuropsychological research, DNMS performance in young animals prominently correlated with the volume of multiple structures in the medial temporal lobe memory system. Less anticipated correlations were also observed in the cingulate and cerebellum. In aged monkeys, significant volumetric correlations with DNMS performance were largely restricted to the prefrontal cortex and striatum. Importantly, interaction effects in an omnibus analysis directly confirmed that the associations between volume and task performance in the MTL and prefrontal cortex are age-dependent. These results demonstrate that the regional distribution of GM volumes coupled with DNMS performance changes across the lifespan, consistent with the perspective that the aged primate brain retains a substantial capacity for structural reorganization.

Background Mosquito larval control through the use of insecticides is the most common strategy for suppressing West Nile virus (WNV) vector populations in Connecticut (CT), United States. To evaluate the ability of larval control to reduce entomological risk metrics associated with WNV, we performed WNV surveillance and assessments of municipal larvicide application programs in Milford and Stratford, CT in 2019 and 2020. Each town treated catch basins and non-basin habitats (Milford only) with biopesticide products during both WNV transmission seasons. Adult mosquitoes were collected weekly with gravid and CO2 -baited light traps and tested for WNV; larvae and pupae were sampled weekly from basins within 500 m of trapping sites, and Culex pipiens larval mortality was determined with lab bioassays of catch basin water samples. Results Declines in 4th instar larvae and pupae were observed in catch basins up to 2-weeks post-treatment, and we detected a positive relationship between adult female Cx. pipiens collections in gravid traps and pupal abundance in basins. We also detected a significant difference in total light trap collections between the two towns. Despite these findings, Cx. pipiens adult collections and WNV mosquito infection prevalence in gravid traps were similar between towns. Conclusion Larvicide applications reduced pupal abundance and the prevalence of host-seeking adults with no detectable impact on entomological risk metrics for WNV. Further research is needed to better determine the level of mosquito larval control required to reduce WNV transmission risk. This article is protected by copyright. All rights reserved.

Background Powassan virus (POWV; genus Flavivirus) is the sole North American member of the tick-borne encephalitis sero-complex and an increasing public health threat in the USA. Maintained in nature by Ixodes spp. ticks, POWV has also been isolated from species of other hard tick genera, yet it is unclear if these species can serve as vectors. Dermacentor variabilis and Amblyomma americanum share geographic and ecologic overlap with Ixodes spp. ticks and POWV transmission foci, raising the possibility that POWV could become established in these tick species and leading to range expansion and increased human risk. Therefore, we assessed the competency of Ixodes scapularis, D. variabilis and A. americanum for POWV lineage II (POWV II). Methods Larvae from all three species were co-infested on POWV-infected Balb/c mice. The engorged larvae were allowed to molt to nymphs and screened for the presence of POWV II RNA by reverse transcription-qPCR. Eight infected nymphs from each species were allowed to individually feed on a naïve mouse. Mice were screened for the presence of POWV II RNA to determine infection status. Results The results demonstrated that larvae from all three tick species were able to efficiently acquire POWV II via feeding on viremic mice, maintain infection through molting and successively transmit POWV to naïve mice at the nymphal stage at comparable rates across all three species. Conclusions Our findings reveal that non-Ixodes tick species can serve as competent vectors for POWV and highlight the potential role of these species in the ecology and epidemiology of POWV. Future studies examining the possible implications of these findings on POWV epidemiology and the adaptability of POWV in these new vectors are warranted. Graphical abstract

La Crosse virus (LACV) is the leading cause of pediatric viral encephalitis in North America, and is an important public health pathogen. Historically, studies involving LACV pathogenesis have focused on lineage I strains, but no former work has explored the pathogenesis between or within lineages. Given the absence of LACV disease in endemic regions where a robust entomological risk exists, we hypothesize that some LACV strains are attenuated and demonstrate reduced neuroinvasiveness. Herein, we compared four viral strains representing all three lineages to determine differences in neurovirulence or neuroinvasiveness using three murine models. A representative strain from lineage I was shown to be the most lethal, causing &gt, 50% mortality in each of the three mouse studies. However, other strains only presented excessive mortality (&gt, 50%) within the suckling mouse neurovirulence model. Neurovirulence was comparable among strains, but viruses differed in their neuroinvasive capacities. Our studies also showed that viruses within lineage III vary in pathogenesis with contemporaneous strains, showing reduced neuroinvasiveness compared to an ancestral strain from the same U.S. state (i.e., Connecticut). These findings demonstrate that LACV strains differ markedly in pathogenesis, and that strain selection is important for assessing vaccine and therapeutic efficacies.

Enzymes are biological catalysts that have many potential industrial and biomedical applications. However, the widespread use of enzymes in the industry has been limited by their instability and poor recovery. In biomedical applications, systemic administration of enzymes has faced two main challenges: limited bioactivity mostly due to rapid degradation by proteases and immunogenic activity, since most enzymes are from nonhuman sources. Herein, we propose a robust enzyme-encapsulation strategy to mitigate these limitations. Catalase (CAT) was encapsulated in nanoporous silica nanoparticles (CAT-SiNPs) by first chemically modifying the enzyme surface with a silica precursor, followed by silica growth and finally poly(ethylene glycol) (PEG) conjugation. The formulation was carried out in mild aqueous conditions and yielded nanoparticles (NPs) with a mean diameter of 230 ± 10 nm and a concentration of 1.3 ± 0.8 × 1012 NPs/mL. CAT-SiNPs demonstrated high enzyme activity, optimal protection from proteolysis by proteinase K and trypsin, and excellent stability over time. In addition, a new electrochemical assay was developed to measure CAT activity in a rapid, simple, and accurate manner without interference from chromophore usually present in biological samples. Concentrations of 2.5 × 1010 to 80 × 1010 CAT-SiNPs/mL not only proved to be nontoxic in cell cultures using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) assay but also conferred cell protection when cells were exposed to 1 mM hydrogen peroxide (H2O2). Finally, the ability of CAT-SiNPs to release oxygen (O2) when exposed to H2O2 was demonstrated in vivo using a rat model. Following the direct injection of CAT-SiNPs in the left kidney, partial pressure of oxygen (pO2) increased by more than 30 mmHg compared to the contralateral control kidney during the systemic infusion of safe levels of H2O2. This pilot study highlights the potential of CAT-SiNPs to generate O2 to relieve hypoxia in tissues and potentially sensitize tumors against radiation therapy.

Campylobacter jejuni is a major cause of foodborne gastroenteritis worldwide inflicting palpable socioeconomic costs. The ability of this pathogen to successfully infect its hosts is determined not only by the presence of specific virulence genes but also by the pathogen’s capacity to appropriately regulate those virulence genes. Therefore, DNA methylation can play a critical role in both aspects of this process because it serves as both a means to protect the integrity of the cellular DNA from invasion and as a mechanism to control transcriptional regulation within the cell. In the present study we report the comparative methylome data of C. jejuni YH002, a multidrug resistant strain isolated from retail beef liver. Investigation into the methylome identified a putative novel motif (CGCGA) of a type II restriction-modification (RM) system. Comparison of methylomes of the strain to well-studied C. jejuni strains highlighted non-uniform methylation patterns among the strains though the existence of the typical type I and type IV RM systems were also observed. Additional investigations into the existence of DNA methylation sites within gene promoters, which may ultimately result in altered levels of transcription, revealed several virulence genes putatively regulated using this mode of action. Of those identified, a flagella gene (flhB), a RNA polymerase sigma factor (rpoN), a capsular polysaccharide export protein (kpsD), and a multidrug efflux pump were highly notable.

Background La Crosse virus (LACV) (genus Orthobunyavirus, family Peribunyaviridae) is a mosquito-borne virus that causes pediatric encephalitis and accounts for 50–150 human cases annually in the USA. Human cases occur primarily in the Midwest and Appalachian regions whereas documented human cases occur very rarely in the northeastern USA. Methods Following detection of a LACV isolate from a field-collected mosquito in Connecticut during 2005, we evaluated the prevalence of LACV infection in local mosquito populations and genetically characterized virus isolates to determine whether the virus is maintained focally in this region. Results During 2018, we detected LACV in multiple species of mosquitoes, including those not previously associated with the virus. We also evaluated the phylogenetic relationship of LACV strains isolated from 2005–2018 in Connecticut and found that they formed a genetically homogeneous clade that was most similar to strains from New York State. Conclusion Our analysis argues for local isolation and long-term persistence of a genetically distinct lineage of LACV within this region. We highlight the need to determine more about the phenotypic behavior of these isolates, and whether this virus lineage poses a threat to public health.

Background: La Crosse virus [LACV] (genus Orthobunyavirus, family Peribunyaviridae) is a mosquito-borne virus that causes pediatric encephalitis and accounts for 50-150 human cases annually in the USA. Human cases occur primarily in the Midwest and Appalachian regions whereas documented human cases occur very rarely in the northeastern USA. Methods: Following detection of a LACV isolate from a field-collected mosquito in Connecticut during 2005, we evaluated the prevalence of LACV infection in local mosquito populations and genetically characterized virus isolates to determine whether the virus is maintained focally in this region. Results: During 2018, we detected LACV in multiple species of mosquitoes, including those not previously associated with the virus. We also evaluated the phylogenetic relationship of LACV strains isolated from 2005-2018 in Connecticut and found that they formed a genetically homogeneous clade that was most similar to strains from New York State. Conclusion: Our analysis argues for local isolation and long-term persistence of a genetically distinct lineage of LACV within this region. We highlight the need to determine more about the phenotypic behavior of these isolates, and whether this virus lineage poses a threat to public health.

Historical declines in multiple insect taxa have been documented across the globe in relation to landscape-level changes in land use and climate. However, declines have either not been universally observed in all regions or examined for all species. Because mosquitoes are insects of public health importance, we analyzed a longitudinal mosquito surveillance data set from Connecticut (CT), United States (U.S.) from 2001 to 2019 to identify changes in mosquito community composition over time. We first analyzed annual site-level collections and metrics of mosquito community composition with generalized linear/additive mixed effects models; we also examined annual species-level collections using the same tools. We then examined correlations between statewide collections and weather variables as well as site-level collections and land cover classifications. We found evidence that the average trap night collection of mosquitoes has increased by ~ 60% and statewide species richness has increased by ~ 10% since 2001. Total species richness was highest in the southern portion of CT, likely due to the northward range expansion of multiple species within the Aedes, Anopheles, Culex, and Psorophora genera. How the expansion of mosquito populations in the northeast U.S. will alter mosquito-borne pathogen transmission in the region will require further investigation.

The Asian tiger mosquito (Aedes albopictus) is an important vector of a number of arboviruses, including Zika (ZIKV), dengue (DENV), and chikungunya (CHIKV) viruses, and has recently expanded its range in the eastern United States to southern New England and New York. Given the recent establishment and proliferation of Ae. albopictus in this region and the increasing amount of international travel between the United States and endemic countries, there is a need to elucidate the public health risk posed by this mosquito species in the Northeast. Accordingly, we evaluated the competence of four Ae. albopictus populations from Connecticut and New York, for two strains each of ZIKV, DENV serotype 2 (DENV-2), and CHIKV, currently circulating in the Americas, to evaluate the local transmission risk by this vector. We found that local Ae. albopictus populations are susceptible to infection by all three viruses but are most capable of transmitting CHIKV. Variation in competence was observed for ZIKV and CHIKV, driven by the virus strains and mosquito population, whereas competence was more homogeneous for the DENV-2 strains under evaluation. These results suggest that under optimal circumstances, Ae. albopictus could support localized transmission of these viruses and emphasize the importance of maintaining mosquito surveillance and control programs to suppress Ae. albopictus populations and limit further range expansion of this species.

Mosquito cell lines have been used extensively in research to isolate and propagate arthropod-borne viruses and understand virus-vector interactions. Despite their utility as an in vitro tool, these cell lines are poorly defined and may harbor insect-specific viruses. Accordingly, we screened four commonly-used mosquito cell lines, C6/36 and U4.4 cells from Aedes albopictus, Aag2 cells from Aedes aegypti, and Hsu cells from Culex quinquefasciatus, for the presence of adventitious (i.e. exogenous) viruses. All four cell lines stained positive for double-stranded RNA, indicative of RNA virus replication. We subsequently identified viruses infecting Aag2, U4.4 and Hsu cell lines using untargeted next-generation sequencing, but not C6/36 cells. PCR confirmation revealed that these sequences stem from active viral replication and/or integration into the cellular genome. Our results show that these commonly-used mosquito cell lines are persistently-infected with several viruses. This finding may be critical to interpreting data generated in these systems. Centers for Disease Control and Prevention [U50/CCU116806-01, U01/CK000509-01]; US Department of Agriculture Hatch Funds and Multistate Research Project [CONH00773, NE1443]; National Institute of Health, National Institute of Allergy and Infectious Diseases [AI067380, AI099042]; NIH/NCATS [UL1 TR001082] This work was supported in part by grants from the Centers for Disease Control and Prevention (U50/CCU116806-01 and U01/CK000509-01), the US Department of Agriculture Hatch Funds and Multistate Research Project (CONH00773 and NE1443), and the National Institute of Health, National Institute of Allergy and Infectious Diseases (AI067380, AI099042), and NIH/NCATS (UL1 TR001082).

Author(s): Power, Melinda C; Mormino, Elizabeth; Soldan, Anja; James, Bryan D; Yu, Lei; Armstrong, Nicole M; Bangen, Katherine J; Delano-Wood, Lisa; Lamar, Melissa; Lim, Yen Ying; Nudelman, Kelly; Zahodne, Laura; Gross, Alden L; Mungas, Dan; Widaman, Keith F; Schneider, Julie | Abstract: ObjectiveOur objectives were to characterize the inter-relation of known dementia-related neuropathologies in one comprehensive model and quantify the extent to which accumulation of neuropathologies accounts for the association between age and dementia.MethodsWe used data from 1,362 autopsied participants of three community-based clinicopathological cohorts: the Religious Orders Study, the Rush Memory and Aging Project, and the Minority Aging Research Study. We estimated a series of structural equation models summarizing a priori hypothesized neuropathological pathways between age and dementia risk individually and collectively.ResultsAt time of death (mean age, 89 years), 44% of our sample had a clinical dementia diagnosis. When considered individually, our vascular, amyloid/tau, neocortical Lewy body, and TAR DNA-binding protein 43 (TDP-43)/hippocampal sclerosis pathology pathways each accounted for a substantial proportion of the association between age and dementia. When considered collectively, the four pathways fully accounted for all variance in dementia risk previously attributable to age. Pathways involving amyloid/tau, neocortical Lewy bodies, and TDP-43/hippocampal sclerosis were interdependent, attributable to the importance of amyloid beta plaques in all three. The importance of the pathways varied, with the vascular pathway accounting for 32% of the association between age and dementia, wheraes the remaining three inter-related degenerative pathways together accounted for 68% (amyloid/tau, 24%; the Lewy body, 1%; and TDP-43/hippocampal sclerosis, 43%).InterpretationAge-related increases in dementia risk can be attributed to accumulation of multiple pathologies, each of which contributes to dementia risk. Multipronged approaches may be necessary if we are to develop effective therapies. Ann Neurol 2018;84:10-22.

According to theory, habitat selection by organisms should reflect underlying habitat-specific fitness consequences and, in birds, reproductive success has a strong impact on population growth in many species. Understanding processes affecting habitat selection also is critically important for guiding conservation initiatives. Northern pintails (Anas acuta) are migratory, temperate-nesting birds that breed in greatest concentrations in the prairies of North America and their population remains below conservation goals. Habitat loss and changing land use practices may have decoupled formerly reliable fitness cues with respect to nest habitat choices. We used data from 62 waterfowl nesting study sites across prairie Canada (1997-2009) to examine nest survival, a primary fitness metric, at multiple scales, in combination with estimates of habitat selection (i.e., nests versus random points), to test for evidence of adaptive habitat choices. We used the same habitat covariates in both analyses. Pintail nest survival varied with nest initiation date, nest habitat, pintail breeding pair density, landscape composition and annual moisture. Selection of nesting habitat reflected patterns in nest survival in some cases, indicating adaptive selection, but strength of habitat selection varied seasonally and depended on population density and landscape composition. Adaptive selection was most evident late in the breeding season, at low breeding densities and in cropland-dominated landscapes. Strikingly, at high breeding density, habitat choice appears to become maladaptive relative to nest predation. At larger spatial scales, the relative availability of habitats with low versus high nest survival, and changing land use practices, may limit the reproductive potential of pintails.

In general, the mammalian whole body mass-specific metabolic rate correlates positively with maximal urine concentration (Umax) irrespective of whether or not the species have adapted to arid or mesic habitat. Accordingly, we hypothesized that the thick ascending limb (TAL) of a rodent with markedly higher whole body mass-specific metabolism than rat exhibits a substantially higher TAL metabolic rate as estimated by Na+-K+-ATPase activity and Na+-K+-ATPase α1-gene and protein expression. The kangaroo rat inner stripe of the outer medulla exhibits significantly higher mean Na+-K+-ATPase activity (~70%) compared with two rat strains (Sprague-Dawley and Munich-Wistar), extending prior studies showing rat activity exceeds rabbit. Furthermore, higher expression of Na+-K+-ATPase α1-protein (~4- to 6-fold) and mRNA (~13-fold) and higher TAL mitochondrial volume density (~20%) occur in the kangaroo rat compared with both rat strains. Rat TAL Na+-K+-ATPase α1-protein expression is relatively unaffected by body hydration status or, shown previously, by dietary Na+, arguing against confounding effects from two unavoidably dissimilar diets: grain-based diet without water (kangaroo rat) or grain-based diet with water (rat). We conclude that higher TAL Na+-K+-ATPase activity contributes to relationships between whole body mass-specific metabolic rate and high Umax. More vigorous TAL Na+-K+-ATPase activity in kangaroo rat than rat may contribute to its steeper Na+and urea axial concentration gradients, adding support to a revised model of the urine concentrating mechanism, which hypothesizes a leading role for vigorous active transport of NaCl, rather than countercurrent multiplication, in generating the outer medullary axial osmotic gradient.

Peripheral blood mononuclear cell (PBMC) gene expression microarray profiling is a minimally invasive tool used in human diet intervention studies. In this study, PBMC gene expression was determined in dogs fed kibble or raw red meat diets for 9 weeks to test the hypothesis that diet influences canine immune cell gene expression profiles. The two diets were associated with differences in PBMC gene expression profiles, which corresponded with changes in plasma IgA concentrations. Analysis of PBMC gene expression profiles might provide useful insights into the long term effects of diet on health outcomes in dogs.

Seasonal abundance of mosquitoes, their viruses, and blood–feeding habits were determined at an open-faced quarry in North Branford, CT, in 2010 and 2011. This unique habitat had not previously been sampled for mosquitoes and mosquito-borne viruses. Thirty species of mosquitoes were identified from 41,719 specimens collected. Coquillettidia perturbans, Aedes trivittatus, and Ae. vexans were the most abundant species and represented 34.5%, 17.7%, and 14.8% of the totals, respectively. Jamestown Canyon virus was isolated from 6 species of mosquitoes collected from mid-June through July: Cq. perturbans (3 pools), Ae. cantator (3), Ae. trivittatus (2), Ae. aurifer (1), Ae. excrucians (1), and Culex pipiens (1). West Nile virus was cultured from 8 pools of Cx. pipiens and from 1 pool of Culiseta melanura collected from mid-August through late September. Cache Valley virus was isolated from 4 species of mosquitoes in 3 genera from about mid-August through late September 2011: Cq. perturbans (5 pools), Ae. trivittatus (2), Anopheles punctipennis (1), and An. quadrimaculatus (1). Nine different mammalian hosts were identified as sources of blood for 13 species of mosquitoes. White-tailed deer, Odocoileus virginianus, were the most common mammalian hosts (90.8%), followed by raccoon, Procyon lotor (3.1%), coyote, Canis latrans (2.4%), and human, Homo sapiens (1.2%). Exclusive mammalian blood-feeding mosquitoes included: Ae. canadensis, Ae. cantator, Ae. excrucians, Ae. japonicus, Ae. vexans, An. punctipennis, and Cx. salinarius. Fourteen species of birds, mostly Passeriformes, were identified as sources of blood from 6 mosquito species. Five species that fed on mammals (Ae. thibaulti, Ae. trivittatus, Ae. cinereus, Cq. perturbans, and Cx. pipiens) also fed on birds.

Background Eastern equine encephalitis virus (EEEV) is an expanding mosquito-borne threat to humans and domestic animal populations in the northeastern United States. Outbreaks of EEEV are challenging to predict due to spatial and temporal uncertainty in the abundance and viral infection of Cs. melanura, the principal enzootic vector. EEEV activity may be closely linked to wetlands because they provide essential habitat for mosquito vectors and avian reservoir hosts. However, wetlands are not homogeneous and can vary by vegetation, connectivity, size, and inundation patterns. Wetlands may also have different effects on EEEV transmission depending on the assessed spatial scale. We investigated associations between wetland characteristics and Cs. melanura abundance and infection with EEEV at multiple spatial scales in Connecticut, USA. Results Our findings indicate that wetland vegetative characteristics have strong associations with Cs. melanura abundance. Deciduous and evergreen forested wetlands were associated with higher Cs. melanura abundance, likely because these wetlands provide suitable subterranean habitat for Cs. melanura development. In contrast, Cs. melanura abundance was negatively associated with emergent and scrub/shrub wetlands, and wetland connectivity to streams. These relationships were generally strongest at broad spatial scales. Additionally, the relationships between wetland characteristics and EEEV infection in Cs. melanura were generally weak. However, Cs. melanura abundance was strongly associated with EEEV infection, suggesting that wetland-associated changes in abundance may be indirectly linked to EEEV infection in Cs. melanura. Finally, we found that wet hydrological conditions during the transmission season and during the fall/winter preceding the transmission season were associated with higher Cs. melanura abundance and EEEV infection, indicating that wet conditions are favorable for EEEV transmission. Conclusions These results expand the broad-scale understanding of the effects of wetlands on EEEV transmission and help to reduce the spatial and temporal uncertainty associated with EEEV outbreaks. Electronic supplementary material The online version of this article (10.1186/s13071-017-2482-0) contains supplementary material, which is available to authorized users.

Creation of grassland communities is a popular approach to conserve grassland-nesting waterfowl and songbirds. Much uncertainty remains regarding the most appropriate seed mixtures and the type and frequency of management. We conducted line-transect songbird surveys in grasslands converted from cropland to native and tame grass-forb mixtures and managed by burning (native only) or haying (native and tame) in Saskatchewan and Manitoba, Canada. Fields were 1–9 years post-management (YPM). Our objective was to improve our understanding of the effects of management regime (native hayed, native burned and tame hayed fields) and frequency (YPM) on passerine density and occurrence in planted grassland. We found limited support for an effect of management regime on passerine density and occurrence in both provinces. Vesper sparrow (Pooecetes gramineus) occurrence in Saskatchewan was greatest in tame hayed fields, and clay-colored sparrow (Spizella pallida) density in Manitoba was greater in tame hayed fields than in native burned fields up to 5 YPM. Years post-management most strongly influenced the occurrence of vesper sparrows in Saskatchewan and sedge wrens (Cistothorus platensis); common yellowthroats (Geothlypis trichas); clay-colored, Savannah (Passerculus sandwichensis), Le Conte's (Ammodramus leconteii), and Nelson's (A. nelsoni) sparrows; and bobolinks (Dolichonyx oryzivorus) in Manitoba. Five of the 8 species influenced by YPM exhibited a year-dependent non-linear response such that they reached their greatest density 4–5 YPM. Our results indicate that the type of stand (native or tame) or management (burning or haying) matters little to the grassland songbird community occupying planted cover fields and that the frequency of management will likely depend upon local environmental conditions in a given region or year. Some form of management of planted grasslands, however, should occur every 4–6 years to maintain habitat for a number of generalist grassland species. © 2017 The Wildlife Society.

Many of the current accredited methods for the molecular detection of Shiga toxin-producing Escherichia coli (STEC) in foods rely on a PCR-based screen for the pathotype-specific genetic markers stx and eae. Unfortunately, these methods can inaccurately conclude the presence of E.coli containing both stx and eae because of the inability of the methods to determine if the two genes originated from a single organism as opposed to a mixture of organisms. This study was undertaken to evaluate if a droplet digital PCR (ddPCR)-based method that does not require DNA isolation could reliably identify the presence of an STEC containing eae in beef samples by confirming that both genes reside within the same cell, even when present in a mixed culture. The ddPCR system used in this study, dd-Check STEC Solution (Bio-Rad), works without the need for DNA isolation by partitioning intact cells into emulsion droplets, where they are lysed, and subsequently undergo multiplexed endpoint PCR. This enables the assay to differentiate between samples where a single organism contains both stx and eae from samples in which stx and eae reside in different organisms. Comparisons were made between the dd-Check STEC Solution, the BAX System Real-Time PCR STEC assay suite (Hygiena), and the iQ-Check STEC PCR detection kit (Bio-Rad) using 37 unique simulations of E. coli contamination in ground beef. While no single platform was consistently superior at detecting eae and stx across all pathogens tested, the results indicated that the dd-Check STEC Solution has the potential to reduce the number of inaccurately identified samples when screening for E. coli with a stx+, eae+ genotype because it can identify the co-existence of multiple virulence genes within a cell even when in the presence of a mixed microbial population containing identical genes. Ultimately, incorporation of this system could result in substantial cost savings by reducing the expenses incurred when product samples are incorrectly classified as containing E. coli with a stx+, eae+ genotype.

Surveillance for diurnal container-inhabiting mosquitoes such as Aedes albopictus (Skuse), Aedes japonicus japonicus (Theobald), and Aedes triseriatus (Say) have routinely relied on the deployment of multiple trap types, including CO2-baited light traps, gravid traps, oviposition traps, and BG-Sentinel. These trap configurations have met with varying degrees of effectiveness and in many instances likely under-sample these key mosquito vectors. Most recently, the BG-Sentinel trap used in conjunction with the human-scent lure has been largely accepted as the gold-standard for monitoring Ae. albopictus. However, its ability to attract other container-inhabiting Aedes species has not been fully evaluated. During 2018, we tested new scent lures, TrapTech Lure-A and Lure-H (Bedoukian Research, Inc.), using BG-Sentinel traps with CO2 in two regions of Connecticut, Stamford and Hamden, against the BG-Lure. Pooled mosquitoes were additionally screened for arbovirus infection. A total of 47,734 mosquitoes representing 8 genera and 32 species were captured during the study, with the Stamford site deriving on average three times as many mosquitoes per trap, adjusting for sampling effort. Lure-A and Lure-H outperformed the BG-Lure in terms of total numbers, diversity evenness, and the proportion of both Ae. j. japonicus and Ae. triseriatus. There were no significant differences among lures in capturing Ae. albopictus, and in terms of species richness. Fifty-seven isolates of virus (West Nile, Jamestown Canyon, and La Crosse viruses) were obtained during the study, with no significant difference between trap-lure. We highlight both novel lures as effective attractants for use in mosquito surveillance=, which either outperform, or equal, BG-Lure.

Bovine lactoferrin is an important milk protein with many health-promoting properties, including improving intestinal barrier integrity. Dysfunction of this barrier, commonly referred to as "leaky gut," has been linked to inflammatory and autoimmune diseases. With some processing techniques, lactoferrin isolated from milk collected at the start of the milking season (early lactation) may have lower purity than that isolated from milk collected during the rest of the milking season (mid-lactation) and could result in differences in bioactivity based on the stage of lactation. We compared reversed-phase HPLC chromatographs of early-lactation and mid-lactation preparations and found that both had large chromatograph peaks at the time predicted for lactoferrin. The notable difference between the 2 chromatographs was a much larger peak in the early-lactation lactoferrin sample that was determined to be angiogenin. Angiogenin was first identified due to its ability to induce new blood vessel formation, but is now known to be involved in numerous physiological processes. Then, we compared the effects of early-lactation and mid-lactation lactoferrin preparations in 2 bioassays: trans-epithelial electrical resistance (TEER), a measure of intestinal barrier integrity, and peripheral blood mononuclear cell cytokine secretion, a measure of immune-stimulatory properties. We found that early-lactation lactoferrin increased TEER across Caco-2 cell layers compared with control from 10 to 48 h, mid-lactation lactoferrin did not alter TEER. We also found that early-lactation lactoferrin reduced the amount of IL-8 produced by peripheral blood mononuclear cells (compared with those treated with control medium) to a greater extent than mid-lactation lactoferrin. A pro-inflammatory chemokine, IL-8 is also known to decrease barrier function. These results suggest that the decrease in IL-8 production in the presence of early-lactation lactoferrin may be the mechanism by which it increases TEER. The anti-inflammatory effect of early-lactation lactoferrin may be related to the presence of angiogenin, which is known to suppress inflammatory responses. This work indicates that products rich in angiogenin may have intestinal health benefits, and further work to investigate this is warranted.

We have developed a set of tools that operate within an aquatic geographic information system to improve the accessibility, and usability of remote-sensed satellite and computer-modeled oceanographic data for marine science and ecosystem-based management. The tools form the Pelagic Habitat Analysis Module (PHAM), which can be applied as a modeling platform, an investigative aid in scientific research, or utilized as a decision support system for marine ecological management. Applications include fisheries, marine biology, physical and biological oceanography, and marine spatial management. The GIS provides a home for diverse data types and automated tools for downloading remote sensed and global circulation model data. Within the GIS environment, PHAM provides a framework for seamless interactive four-dimensional visualization, for matching between disparate data types, for flexible statistic or mechanistic model development, and for dynamic application of user developed models for habitat, density, and probability predictions. Here we describe PHAM in the context of ecosystem-based fisheries management, and present results from case study projects which guided development. In the first, an analysis of the purse seine fishery for tropical tuna in the eastern Pacific Ocean revealed oceanographic drivers of the catch distribution and the influence of climate-driven circulation patterns on the location of fishing grounds. To support management of the Common Thresher Shark (Alopias vulpinus) in the California Current Ecosystem, a simple empirical habitat utilization model was developed and used to dynamically predict the seasonal range expansion of common thresher shark based on oceanographic conditions.

The vertebrate fauna of the Sir Joseph Banks Group includes 6 native (2 now extinct) and 7 introduced mammals, 76 native and 4 introduced birds and 15 native and 1 introduced reptile. The determination of the total mammal fauna includes examination of sub-fossil bone collections from both Reevesby and Spilsby Islands. Tammar wallaby, bush rat and greater stick-nest rats were only recorded from sub-fossils, but the successful reintroduction of the stick-nest rat to Reevesby Island begun in 1990, is described. Two species of bat are recorded from the two large islands, Spilsby and Reevesby. An Australian sea lion haul-out site on Blyth Island is described for the first time. Counts of the number of pairs of Cape Barren geese now breeding on Spilsby Island are also published here for the first time. The islands of the Sir Joseph Banks Group are very significant for the conservation of species with large and important breeding populations there such as Cape Barren geese, black-faced cormorants, white-...

Assessment of stress from varied sources is usually evaluated using individuals derived from a single population and is assumed to represent all populations of that species. However, recent research has identified intraspecies variations in the stress response, which may be mediated by life history. We examined how life history can influence the physiological responses to an acute stress event by evaluating sea lamprey (Petromyzon marinus) parasitism response in two lake trout (Salvelinus namaycush) morphotypes: the lean and the siscowet. The morphotypes differ in that the lean grows faster, is more fecund, and has a shorter life span. In contrast, the siscowet grows slower, is older at maturity, and lives longer. Our study compared long-term parasitism responses between wild leans and siscowets in Lake Superior and immediate responses in laboratory parasitism trials using lake trout reared under common environmental conditions. Leans, but not siscowets, showed changes in steroid-binding protein function and weak evidence of gigantism in response to parasitism. Both morphotypes showed indications of reproductive endocrine alterations in response to parasitism. Our results demonstrate intraspecies variation in physiological stressor response, which is mediated by life history differences that could potentially have differential population implications.

Ticks transmit a diverse array of viruses such as tick-borne encephalitis virus, Powassan virus, and Crimean-Congo hemorrhagic fever virus that are reemerging in many parts of the world. Most tick-borne viruses (TBVs) are RNA viruses that replicate using error-prone polymerases and produce genetically diverse viral populations that facilitate their rapid evolution and adaptation to novel environments. This article reviews the mechanisms of virus transmission by tick vectors, the molecular evolution of TBVs circulating in nature, and the processes shaping viral diversity within hosts to better understand how these viruses may become public health threats. In addition, remaining questions and future directions for research are discussed.

Citrus canker, caused by Xanthomonas citri ssp. citri, is a serious disease that causes substantial economic losses to the citrus industry worldwide. The bactericide bismerthiazol has been used to control rice bacterial blight (X. oryzae pv. oryzae). In this paper, we demonstrate that bismerthiazol can effectively control citrus canker by both inhibiting the growth of X. citri ssp. citri and triggering the plant’s host defense response through the expression of several pathogenesis-related genes (PR1, PR2, CHI, and RpRd1) and the nonexpresser of PR genes (NPR1, NPR2, and NPR3) in ‘Duncan’ grapefruit, especially at early treatment times. In addition, we found that bismerthiazol induced the expression of the marker genes CitCHS and CitCHI in the flavonoid pathway and the PAL1 (phenylalanine ammonia lyase 1) gene in the salicylic acid (SA) biosynthesis pathway at different time points. Moreover, bismerthiazol also induced the expression of the priming defense-associated gene AZI1. Taken together, these results indicate that the induction of the defense response in ‘Duncan’ grapefruit by bismerthiazol may involve the SA signaling pathway and the priming defense and that bismerthiazol may serve as an alternative to copper bactericides for the control of citrus canker.

The genus Flavivirus includes a number of newly recognized viruses that infect and replicate only within mosquitoes. To determine whether insect-specific flaviviruses (ISFs) may infect Culiseta (Cs.) melanura mosquitoes, we screened pools of field-collected mosquitoes for virus infection by RT-PCR targeting conserved regions of the NS5 gene. NS5 nucleotide sequences amplified from Cs. melanura pools were genetically similar to other ISFs and most closely matched Calbertado virus from Culex tarsalis, sharing 68.7% nucleotide and 76.1% amino acid sequence identity. The complete genome of one virus isolate was sequenced to reveal a primary open reading frame (ORF) encoding a viral polyprotein characteristic of the genus Flavivirus. Phylogenetic analysis showed that this virus represents a distinct evolutionary lineage that belongs to the classical ISF group. The virus was detected solely in Cs. melanura pools, occurred in sampled populations from Connecticut, New York, New Hampshire, and Maine, and infected both adult and larval stages of the mosquito. Maximum likelihood estimate infection rates (MLE-IR) were relatively stable in overwintering Cs. melanura larvae collected monthly from November of 2012 through May of 2013 (MLE-IR = 0.7-2.1/100 mosquitoes) and in host-seeking females collected weekly from June through October of 2013 (MLE-IR = 3.8-11.5/100 mosquitoes). Phylogenetic analysis of viral sequences revealed limited genetic variation that lacked obvious geographic structure among strains in the northeastern United States. This new virus is provisionally named Culiseta flavivirus on the basis of its host association with Cs. melanura.

Understanding factors that influence the demography of grassland species in agricultural landscapes is essential given the amount of land under agricultural production. Winter wheat is promoted by avian conservation organizations in North America because it provides cover early in the breeding season without much disturbance to nesting waterfowl. We conducted point-count surveys in fields of winter wheat, spring wheat, summerfallow, planted grassland and native grassland to determine: 1) the relative benefits of winter wheat to songbirds, 2) the extent to which cropland suitability changes over the breeding season, and 3) the extent to which the surrounding landscape influences songbird abundance. Horned lark (Eremophila alpestris), Savannah sparrow (Passerculus sandwichensis) and vesper sparrow (Pooecetes gramineus) were most commonly recorded in wheat and summerfallow fields, and their abundance varied over the breeding season and with the amount of native grassland in the surrounding landscape. Horned lark was more abundant in winter wheat early in the breeding season and its abundance increased with the amount of native grassland in the surrounding landscape. Savannah sparrow abundance increased in all field-types over the breeding season and decreased in cropland fields as the amount of native grassland in the surrounding landscape increased. Programs that encourage or incentivize winter wheat seeding in grassland-dominated landscapes will yield the greatest benefits for horned lark whereas Savannah sparrows would derive the greatest benefit in cropland-dominated landscapes. However, such programs must target cultivated fields to ensure that grassland obligate species are not compromised.

Mycobacterium tuberculosis is an acid-fast intracellular pathogen of macrophages and the etiological agent of the human respiratory disease tuberculosis (TB). A key aspect of the M. tuberculosis lifecycle is its ability to establish long-term persistent (i.e. latent) infection within the lungs of infected individuals. During latency, M. tuberculosis produces, stores, and utilizes lipid droplets containing the neutral fatty acid triacylglycerol. In many organisms including M. tuberculosis, lipid droplets have been shown to function as a long-term energy storage depots, providing energy for key metabolic pathways, and serving as a carbon source for the generation of important biosynthetic precursors. Failure of M. tuberculosis to generate, maintain, or utilize triacylglycerol stored within lipid droplets leads to reduced survival of the bacterium in in vitro and in vivo models of persistence. These findings implicate lipid droplets as central organelles vital to the physiology and pathogenesis of M. tuberculosis within the host. In this chapter, the persistence of M. tuberculosis in the context of lipid droplet biogenesis, storage, and utilization is discussed.

West Nile virus (WNV; Flaviviridae: Flavivirus) is a widely distributed arthropod-borne virus that has negatively affected human health and animal populations. WNV infection rates of mosquitoes and human cases have been shown to be correlated with climate. However, previous studies have been conducted at a variety of spatial and temporal scales, and the scale-dependence of these relationships has been understudied. We tested the hypothesis that climate variables are important to understand these relationships at all spatial scales. We analyzed the influence of climate on WNV infection rate of mosquitoes and number of human cases in New York and Connecticut using Random Forests, a machine learning technique. During model development, 66 climate-related variables based on temperature, precipitation and soil moisture were tested for predictive skill. We also included 20-21 non-climatic variables to account for known environmental effects (e.g., land cover and human population), surveillance related information (e.g., relative mosquito abundance), and to assess the potential explanatory power of other relevant factors (e.g., presence of wastewater treatment plants). Random forest models were used to identify the most important climate variables for explaining spatial-temporal variation in mosquito infection rates (abbreviated as MLE). The results of the cross-validation support our hypothesis that climate variables improve the predictive skill for MLE at county- and trap-scales and for human cases at the county-scale. Of the climate-related variables selected, mean minimum temperature from July-September was selected in all analyses, and soil moisture was selected for the mosquito county-scale analysis. Models demonstrated predictive skill, but still over- and under-estimated WNV MLE and numbers of human cases. Models at fine spatial scales had lower absolute errors but had greater errors relative to the mean infection rates.

An animal's ability to navigate an olfactory environment is critically dependent on the activities of its first-order olfactory receptor neurons (ORNs). While considerable research has focused on ORN responses to odorants, the mechanisms by which olfactory information is encoded in the activities of ORNs and translated into navigational behavior remain poorly understood. We sought to determine the contributions of most Drosophila melanogaster larval ORNs to navigational behavior. Using odorants to activate ORNs and a larval tracking assay to measure the corresponding behavioral response, we observed that larval ORN activators cluster into four groups based on the behavior responses elicited from larvae. This is significant because it provides new insights into the functional relationship between ORN activity and behavioral response. Subsequent optogenetic analyses of a subset of ORNs revealed previously undescribed properties of larval ORNs. Furthermore, our results indicated that different temporal patterns of ORN activation elicit different behavioral outputs: some ORNs respond to stimulus increments while others respond to stimulus decrements. These results suggest that the ability of ORNs to encode temporal patterns of stimulation increases the coding capacity of the olfactory circuit. Moreover, the ability of ORNs to sense stimulus increments and decrements facilitates instantaneous evaluations of concentration changes in the environment. Together, these ORN properties enable larvae to efficiently navigate a complex olfactory environment. Ultimately, knowledge of how ORN activity patterns and their weighted contributions influence odor coding may eventually reveal how peripheral information is organized and transmitted to subsequent layers of a neural circuit.