The Operophtera brumata Nucleopolyhedrovirus (OpbuNPV) Represents an Early, Divergent Lineage within Genus Alphabaculovirus

Operophtera brumata nucleopolyhedrovirus (OpbuNPV) infects the larvae of the winter moth, Operophtera brumata. As part of an effort to explore the pesticidal potential of OpbuNPV, an isolate of this virus from Massachusetts (USA)—OpbuNPV-MA—was characterized by electron microscopy of OpbuNPV occlusion bodies (OBs) and by sequencing of the viral genome. The OBs of OpbuNPV-MA consisted of irregular polyhedra and contained virions consisting of a single rod-shaped nucleocapsid within each envelope. Presumptive cypovirus OBs were also detected in sections of the OB preparation. The OpbuNPV-MA genome assembly yielded a circular contig of 119,054 bp and was found to contain little genetic variation, with most polymorphisms occurring at a frequency of \u3c 6%. A total of 130 open reading frames (ORFs) were annotated, including the 38 core genes of Baculoviridae, along with five homologous repeat (hr) regions. The results of BLASTp and phylogenetic analysis with selected ORFs indicated that OpbuNPV-MA is not closely related to other alphabaculoviruses. Phylogenies based on concatenated core gene amino acid sequence alignments placed OpbuNPV-MA on a basal branch lying outside other alphabaculovirus clades. These results indicate that OpbuNPV-MA represents a divergent baculovirus lineage that appeared early during the diversification of genus Alphabaculovirus

Insights Into the Feeding Behaviors and Biomechanics of \u3ci\u3eVarroa destructor\u3c/i\u3e Mites on Honey Bee Pupae Using Electropenetrography and Histology

Feeding behaviors and biomechanics of female Varroa destructor mites are revealed from AC-DC electropenetrography (EPG) recordings of mites feeding from Apis mellifera honey bee pupae and histology of mite internal ingestion apparatus. EPG signals characteristic of arthropod suction feeding (ingestion) were identified for mites that fed on pupae during overnight recordings. Ingestion by these mites was confirmed afterwards by observing internally fluorescent microbeads previously injected into their hosts. Micrographs of internal ingestion apparatus illustrate the connection between a gnathosomal tube and a pharyngeal lumen, which is surrounded by alternating dilator and constrictor muscles. Inspection of EPG signals showed the muscularized mite pharyngeal pump operates at a mean repetition rate of 4.5 cycles/s to ingest host fluids. Separate feeding events observed for mites numbered between 23 and 33 over approximately 16 h of recording, with each event lasting ~10 s. Feeding events were each separated by ~2 min. Consecutive feeding events separated by either locomotion or prolonged periods of quiescence were grouped into feeding bouts, which ranged in number from one to six. Statistical analyses of EPG data revealed that feeding events were prolonged for mites having lower pharyngeal pump frequencies, and mites having prolonged feeding events went unfed for significantly more time between feeding events. These results suggest that mites may adjust behaviors to meet limitations of their feeding apparatus to acquire similar amounts of food. Data reported here help to provide a more robust view of Varroa mite feeding than those previously reported and are both reminiscent of, as well as distinct from, some other acarines and fluid-feeding insects

A new species of Zambedania (Acari : Heterostigmatina : Pygmephoridae) from the two rivers platinum mine in South Africa and notes on the life-cycle of the genus

A new species of relatively poorly known genus Zambedania Mahunka, 1972 was found on the baboon spider, Harpactirella overdijki Gallon, 2010 (Araneae: Theraphosidae) in South Africa. Besides the abundantly available phoretic females, several males and one larva of this species in the spiders’ nests were also collected. Zambedania sekhukhunensis n. sp. is described and illustrated based on the phoretic females, males and larva. Improved diagnosis of the genus and a new key to species are also supplied. The descriptions and illustrations of the male and larva of this species represent the first ones of these stages in the genus Zambedania. Due to their discovery the generic diagnosis has been significantly improved.This paper is part of a South Africa/ Poland Research Cooperation Programme.The National Research Foundation of South Africa through the University of the Witwatersrand and Polish Ministry of Science and Higher Education through the A. Mickiewicz University (Poznań).http://www.zoologicalstudies.comam2017Zoology and Entomolog

Scientific Works of Art Reveal a Hidden World

It has been said that a picture is worth a thousand words, and at the Agricultural Research Service’s Electron and Confocal Microscopy Unit (ECMU) in Beltsville, Maryland, this adage couldn’t be more true. Led by unit director Gary Bauchan, the ECMU is tasked with producing highresolution images that provide a window to the extraordinary world of the unseen. “We have observed viruses, bacteria, fungi, nematodes, insects, mites, and parasites that threaten global food security, and we’ve contributed to the discovery of how pathogens spread by helping elucidate their relationship to the environment, hosts, and vectors,” says Bauchan. “We’ve also described new biocontrol agents for the management of pathogens and characterized healthy and infected plant and animal tissues to discern the structural changes caused by pathogens.” Other ECMU capabilities include tracking the development of genetically transformed plants using fluorescently tagged plant cells and tissues and contributing to improved food safety by determining the mechanisms by which bacteria, fungi, and parasites infect fresh produce

Genetic transformation of \u3ci\u3eFusarium oxysporum\u3c/i\u3e f.sp. \u3ci\u3egladioli\u3c/i\u3e with \u3ci\u3eAgrobacterium\u3c/i\u3e to study pathogenesis in \u3ci\u3eGladiolus\u3c/i\u3e

Fusarium rot caused by Fusarium oxysporum f.sp. gladioli (Fog) is one of the most serious diseases of Gladiolus, both in the field and in bulbs in storage. In order to study the mechanisms of pathogenesis of this fungus, we have transformed Fog with Agrobacterium tumefaciens binary vectors containing the hygromycin B phosphotransferase (hph) gene and fluorescence reporter genes EGFP (green), EYFP (yellow) or ECFP (cyan) using the AGL-1 strain of A. tumefaciens. Hygromycin B (100 μg/ml) resistant colonies were observed only when acetosyringone was added to the co-cultivation medium. Transformed colonies are more clearly visible when co-cultivated on cellophane membrane than on Hybond -N+ membrane. Transformed lines were stably maintained through four serial passages on medium containing hygromycin B, and they expressed green, yellow or cyano fluorescence. PCR with hph-specific primers and Southern blotting with an hph-specific probe were positive for HygR lines but not for the untransformed isolate. The cyano fluorescence of the ECFP-transformed isolate was clearly distinguishable from the green autofluorescence of Gladiolus roots, signifying the potential of these lines for further histopathological investigations. Transformed lines will be useful for identifying pathogenicity related genes, screening transgenic resistance, and in studies of host-pathogen interactions

Spinacia oleracea L. Leaf Stomata Harboring Cryptosporidium parvum Oocysts: a Potential Threat to Food Safety ▿ †

Cryptosporidium parvum is a cosmopolitan microscopic protozoan parasite that causes severe diarrheal disease (cryptosporidiosis) in mammals, including humans and livestock. There is growing evidence of Cryptosporidium persistence in fresh produce that may result in food-borne infection, including sporadic cases as well as outbreaks. However, drinking and recreational waters are still considered the major sources of Cryptosporidium infection in humans, which has resulted in prioritization of studies of parasite etiology in aquatic environments, while the mechanisms of transmission and parasite persistence on edible plants remain poorly understood. Using laser scanning confocal microscopy together with fluorescein-labeled monoclonal antibodies, C. parvum oocysts were found to strongly adhere to spinach plants after contact with contaminated water, to infiltrate through the stomatal openings in spinach leaves, and to persist at the mesophyll level. These findings and the fact that this pathogenic parasite resists washing and disinfection raise concerns regarding food safety

Magdalenapalpus Mesa, Welbourn and Evans 2009

<i>Magdalenapalpus</i> Mesa, Welbourn and Evans, 2009 <p> <b>Type species.</b> <i>Meyeraepalpus strandtmanni</i> Smiley, Frost and Gerson, 1996, by original designation.</p> <p> <b>Diagnosis.</b> <i>All life stages:</i> dorsal opisthosoma with 12 or 13 pairs of lanceolate setae; <i>c2</i>, <i>d2</i>, and <i>e2</i> present; seta <i>f2</i> present or absent; setae <i>e2</i> inserted in more-or-less marginal position; setae <i>h2</i> similar in size and form to other dorsal setae; palps 5-segmented, setal formula 0,0,0,2,3(1); immature stages with anterior margin of prodorsum smoothly rounded, without projections/notches, gnathosoma not concealed; ventral plate absent; 3 pairs of pseudanal setae (<i>ps1–3</i>) on weakly developed anal plates. <i>Adult female:</i> anterior margin of prodorsum deeply incised, forming 1 pair of broad fleshy lobes, each lobe bearing setae <i>v2</i> (also in male); gnathosoma partially concealed by anterior margin of prodorsum (also in male); genital plate weakly developed, membranous; metapodal plates not developed; coxae I without setae <i>1c</i>; trochanters I–IV 0-0-1-0 (<i>v ′</i> absent on tr I–IV; <i>l ′</i> present on tr III); femora I–IV 3-3 -2-1; genua 1-1-0-0 (<i>d</i> present on ge I–II); tibiae 4-4-3-3; tarsi I–IV 8 (1)-8(1)-4-4 (without <i>tc′′</i>). Solenidia of male much thicker and longer than in female.</p> <p> <b>Species.</b> Three species: <i>M. strandtmanni</i>, <i>M. caperatus</i>, <i>M. forsteri</i>.</p> <p> <b>Hosts and distribution.</b> Casuarinaceae, Australia.</p> <p> <b>Remarks.</b> The new species described herein differ from the type species by lacking seta <i>f2</i>. Although this is an important difference, these species share the same leg setation, a deeply incised prodorsum with setae <i>v2</i> inserted on the resultant lobes, and a partially concealed gnathosoma.</p>Published as part of <i>Beard, Jennifer J., Seeman, Owen D. & Bauchan, Gary R., 2014, Tenuipalpidae (Acari: Trombidiformes) from Casuarinaceae (Fagales), pp. 1-157 in Zootaxa 3778 (1)</i> on page 51, DOI: 10.11646/zootaxa.3778.1.1, <a href="http://zenodo.org/record/251337">http://zenodo.org/record/251337</a&gt

Pentamerismus wardo Seeman and Beard, sp. nov.

<i>Pentamerismus wardo</i> Seeman and Beard sp. nov. <p>(Figs 82–87)</p> <p> <b>Type material examined.</b> Holotype female ex. <i>Allocasuarina scleroclada</i> (Casuarinaceae), <b>AUSTRALIA:</b> Western Australia, approx. 3 km N Coomberdale, along Moora-Watheroo Road, 30°24’17” S 116°02’29” E, 15 April 2009, coll. J.J. Beard (WAM) (BRI voucher, BRI [AQ814925]). Paratypes. 6 females, 3 pharate females, 2 males, 2 pharate males, 4 larvae, same data as holotype (WAM, QM, ANIC, USNM).</p> <p> <b>Diagnosis.</b> Dorsal opisthosomal setae <i>f2</i> present. Palp setal formula 0-0-0-2-3(1), with palp tibial setae <i>l’PTi</i> and <i>l′′PTi</i> both present. Anterior margin of prodorsal shield with short notch. Dorsal shields with irregular weakly reticulate to rugose mosaic sculpturing; dorsal setae broadly lanceolate, barbed; lateral margins of opisthosoma with transverse to oblique folds or striae. Posterior ventral setae <i>ag</i> fine, barbed; setae <i>g1–2</i> fine, barbed; setae <i>ps1– 3</i> fine, <i>ps1</i> slightly thicker than <i>ps2–3</i>, <i>ps1–2</i> barbed, <i>ps3</i> smooth. Seta <i>1c</i> absent. Setae <i>d</i> on femora and genua I–II lanceolate; seta <i>ev ′</i> on femora III fine, smooth; setae <i>v ′, v′′</i> on tibiae III fine, smooth to weakly barbed.</p> <p> FEMALE (n = 7). <i>Dorsum.</i> (Fig. 82 a) Body measurements: distance between setae <i>v2 -h1</i> 220–260 [260], <i>sc2- sc2</i> 105–120 [110]; other measurements: <i>v2-v2</i> 40 –46 [45], <i>sc1-sc1</i> 77–86 [86], <i>c1-c1</i> 49–72 [51], <i>c2-c2</i> 110–135 [110], <i>c3-c3</i> 160–180 [160], <i>d1-d1</i> 33–38 [38], <i>d2-d2</i> 90–115 [98], <i>d3-d3</i> 130–150 [135], <i>e1- e1</i> 36–43 [41], <i>e2-e2</i> 120–140 [130], <i>e3-e3</i> 110–125 [115], <i>f2-f2</i> 96–110 [100], <i>f3-f3</i> 77–89 [80], <i>h1-h 1</i> 24–29 [24], <i>h2-h2</i> 54–63 [54].</p> <p> Anterior margin of prodorsal shield with 1 pair of small lobes forming short medial notch (internal depth 9–12). Prodorsal shield with irregular weakly reticulate to rugose mosaic of polygonal cells medially, with cells elongate anteriorly and laterally. Opisthosomal shield with similar pattern to prodorsal shield, except pattern less complete, especially laterally. Lateral cuticle surrounding prodorsal shield smooth to weakly rugose medially, with some weak folds posteriorly; lateral cuticle surrounding opisthosoma with transverse to oblique folds and striae. All dorsal setae barbed, lanceolate, becoming broader posteriorly. Setal lengths: <i>v 2</i> 22–27 [22], <i>sc 1</i> 24–27 [24], <i>sc 2</i> 26–31 [26], <i>c 1</i> 22–26 [22], <i>c 2</i> 25–26 [25], <i>c 3</i> 20–23 [20], <i>d 1</i> 16–17 [not measurable], <i>d 2</i> 16–20 [16], <i>d 3</i> 18–20 [18], <i>e 1</i> 15 –17 [not measurable], <i>e 2</i> 17 –19 [17], <i>e 3</i> 19 –21 [19], <i>f 2</i> 17–19 [17], <i>f 3</i> 17–19 [17], <i>h 1</i> 17–18 [17], <i>h 2</i> 18– 20 [18]. <i>Palps</i>. (Fig. 82 b) Setal formula 0, 0, 0, 2, 3(1s+2e). Tibial setae, dorsal 5–6 [6] long, ventral 7–9 [7] long; tarsal eupathidia 3–4 [3] long, 5 [5] long; solenidion 5–6 [5] long. <i>Venter.</i> (Fig. 83) Cuticle with transverse striae, becoming longitudinal just anterior to setae <i>ag</i>, becoming coarse around genital area. Circular thickening present in metapodal region, ca. 25 diameter, rugose. Setae <i>g1</i> inserted in more-or-less transverse line with <i>g2</i>, <i>g2</i> slightly anterior to <i>g1</i>. Genital shield punctate, margins irregular, 30–32 [32] long, 40–45 [43] wide; anal setae <i>ps1–3</i> inserted in longitudinal row on anal plates. Coxal setae fine, except <i>2c</i> thickened, barbed; setae <i>ag1</i> fine, barbed; <i>g1–2</i>, <i>ps1–2</i> fine, barbed; <i>ps3</i> fine, smooth. Setal lengths: <i>1a</i> 47–52 [47], <i>1b</i> 18–31 [17], <i>2b</i> 20–25 [20], <i>2c</i> 16–20 [17], <i>3a</i> 49–58 [49], <i>3b</i> 15–19 [19], <i>4a</i> 46–55 [46], <i>4b</i> 11–17 [15], <i>ag 1</i> 12–20 [19], <i>g 1</i> 18–24 [20], <i>g 2</i> 15–22 [15], <i>ps 1</i> 13–15 [13], <i>ps 2</i> 13–14 [13], <i>ps3</i> 7–8 [17]. <i>Spermatheca</i>. Spermathecal tube long, narrow, convoluted, <1 wide,> 70 long. Spermatheca vesicle obscured in all specimens. Genital opening between anal valves and posterior margin of genital shield. <i>Legs.</i> (Fig. 84) Setal formula for legs I–IV (coxae to tarsi) 1-1-3-3-4-9(1), 2-1-3-3-4-9(1), 1-2-2-1-3-5, 1-1-1-0-3-5. Tarsi I and II each with 1 antiaxial solenidion <i>ω"</i> (9–10 [9] long) and 2 eupathidia <i>pζ'-pζ"</i> (6–7 [7] long). Leg setation as in Table 1 except coxae I without <i>1c.</i></p> <p> MALE (n = 2). <i>Dorsum.</i> (Fig. 85) Body measurements: distance between setae <i>v2 -h1</i> 185–190, <i>sc2-sc2</i> 85–88; other measurements: <i>v2-v 2</i> 26–30, <i>sc1-sc1</i> 63–67, <i>c1-c1</i> 39–41, <i>c2-c2</i> 89–93, <i>c3-c3</i> 120–125, <i>d1-d 1</i> 20–22, <i>d2-d2</i> 70–74, <i>d3-d3</i> 93–97, <i>e1- e 1</i> 29–34, <i>e2- e2</i> 89–93, <i>e3- e3</i> 17, <i>f2-f2</i> 76–77, <i>f3-f3</i> 61–63, <i>h1-h 1</i> 13–14, <i>h2-h2</i> 15. Anterior margin of prodorsum with pair of small lobes forming a short medial notch (internal depth 3–4). Prodorsal, mesonotal and pygidial shields with sculpture and dorsal setae similar to female. Medial soft cuticle striated, lateral cuticle smooth, becoming festo1d posteriorly. Setal lengths: <i>v 2</i> 20–22, <i>sc1</i> 21, <i>sc 2</i> 21–22, <i>c1</i> 18, <i>c 2</i> 19–21, <i>c3</i> 17, <i>d 1</i> 11–13, <i>d2</i> 15, <i>d 3</i> 15–16, <i>e1</i> 12, <i>e2</i> 16, <i>e3</i> 17, <i>f2</i> 16, <i>f 3</i> 16–17, <i>h 1</i> 13–14, <i>h2</i> 15. <i>Palps</i>. (Fig. 85) Palps similar to female. Tibial setae, dorsal 6 long, ventral 7–8 long; tarsal eupathidia 3, 5–6 long; solenidion 4–5 long. <i>Venter.</i> (Fig. 86) Striae entirely transverse, becoming coarse behind cx IV. Posterior opisthosoma with 2 irregular, poorly defined, striated subcircular plates, ca. 30 in diameter; <i>g1–2</i>, <i>ps1–3</i> on weakly sclerotised anal valves; <i>ps1</i> modified, thickened. Coxal setae fine, except <i>2c</i> thickened, barbed. Seta <i>ag1</i> narrowly lanceolate, barbed; <i>g1</i> lanceolate; <i>g2</i>, <i>ps2, ps3</i> smooth; <i>ps1</i> spine-like, thickened. Setal lengths: <i>1a</i> 38–48, <i>1</i> b 26, <i>2</i> b 26, <i>2</i> c 13, <i>3a</i> 30–37, <i>3</i> b 18–19, <i>4</i> a 47, <i>4</i> b 20, <i>ag1</i> 13, <i>g1</i> 11, <i>g 2</i> 11–13, <i>ps 1</i> 11–14, <i>ps2</i> 7, <i>ps3</i> 6–7. <i>Aedeagus</i>. Narrow, sclerotised, tapering to a point, 66 long, curved. Membranous duct runs from inside aedeagus. <i>Legs.</i> (Fig. 85) Setal formula same as female. Tarsi I and II each with 1 antiaxial solenidion <i>ω"</i> (9–10 long) and 2 eupathidia <i>pζ'-pζ"</i> (about 6 long).</p> <p> LARVA (n = 2). <i>Dorsum.</i> (Fig. 87) Body measurements: distance between setae <i>v2 -h1</i> 130–140, <i>sc2-sc2</i> 54– 59; other measurements: <i>v2-v 2</i> 20–22, <i>sc1-sc1</i> 44–48, <i>c1-c 1</i> 17–20, <i>c2-c2</i> 61, <i>c3-c3</i> 87–89, <i>d1-d1</i> 13, <i>d2-d2</i> 51–52, <i>d3-d3</i> 72–76, <i>e1- e 1</i> 8–10, <i>e2- e2</i> 56–61, <i>e3- e3</i> 50–53, <i>f2-f2</i> 36–38, <i>f3-f 3</i> 24–31, <i>h1-h1</i> 4–6, <i>h2-h 2</i> 11–16. Anterior margin of prodorsum without medial notch. Prodorsal shield 54–57 long, 64 wide, defined by numerous longitudinal striations. Opisthosomal shield absent; coarse, irregular transverse striae, becoming obsolete between <i>d1-e1</i>. Setae similar in length to those of adult; setae narrowly lanceolate to thickened, barbed; setae <i>d1</i> and <i>e1</i> broadest; setae <i>v2, sc1, sc2, h2</i> narrowest. Setal lengths: <i>v 2</i> 14–19, <i>sc 1</i> 13–15, <i>sc 2</i> 14–17, <i>c 1</i> 19–20, <i>c2</i> 14, <i>c 3</i> 14– 15, <i>d 1</i> 19–20, <i>d 2</i> 17–18, <i>d 3</i> 17–20, <i>e 1</i> 21 –22, <i>e 2</i> 20 –21, <i>e 3</i> 20 –23, <i>f2</i> 20, <i>f 3</i> 24–26, <i>h 1</i> 24–25, <i>h2</i> 20. <i>Palps</i>. (Fig. 87) Palps similar to adult. Tibial setae, dorsal 4 long, ventral 5 long; tarsal eupathidia 3, 4 long; solenidion 3 long. <i>Vent er.</i> Cuticle with transverse striae to setae <i>3a</i>, oblique posterior to <i>3a,</i> transverse to midway between <i>3a</i> and <i>ps3</i>, then longitudinal anal area, slightly coarser around anal area. Anal setae <i>ps1–3</i> on weakly defined anal plates. Coxal setae fine. Setal lengths: <i>1a</i> 25–30, <i>1</i> b 15, <i>3a</i> 35–38, <i>ps1</i> 3–4, <i>ps2</i> 4–5, <i>ps3</i> 4–5. <i>Legs.</i> (Fig. 87) Setal formula for legs I–III (coxae to tarsi) 1-0-3-1-4-7(1), 0-0-3-1-4-7(1), 0-0-2-1-3-3. Tarsi I and II each with 1 antiaxial solenidion <i>ω"</i> (ta I 5 long, ta II 4 long) and 2 eupathidia <i>pζ'-pζ"</i> (about 4 long). Leg setation as in Table 1.</p> <p> <b>Etymology.</b> The specific name, <i>wardo</i>, is an Australian indigenous word meaning “a little bird” referring specifically to the Willy Wagtail (<i>Rhipidura leucophrys</i> Latham (Rhipiduridae)), and is the source of the town name Watheroo, near which this mite was collected.</p> <p> <b>Remarks.</b> <i>Pentamerismus wardo</i> is similar to <i>P. hicklingorum</i>, but can be separated by having two setae on the palp tibia (one seta on the palp tibia of <i>P. hicklingorum</i>) and the dorsal cuticle being weakly reticulate (with broadly rounded folds on <i>P. hicklingorum</i>).</p> <p>Individuals were found buried in the thick pubescence present on the tips of the needle-like stems and branchlets of the host.</p>Published as part of <i>Beard, Jennifer J., Seeman, Owen D. & Bauchan, Gary R., 2014, Tenuipalpidae (Acari: Trombidiformes) from Casuarinaceae (Fagales), pp. 1-157 in Zootaxa 3778 (1)</i> on pages 97-104, DOI: 10.11646/zootaxa.3778.1.1, <a href="http://zenodo.org/record/251337">http://zenodo.org/record/251337</a&gt