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Biomedical subjects

W P Wergin

Publications and source records attributed to W P Wergin.

At least 19 recordsLinked to original sources

Development of Onchocerca volvulus (Filarioidea: Onchocercidae) in the West African black fly Simulium yahense (Diptera: Simuliidae) in Liberia.

Simulium yahense black flies infected with microfilaria of Onchocerca volvulus were kept in a defined insectary environment in Liberia, West Africa. A daily sample of infected flies was dissected for larvae developing in the thoracic muscles and examined for growth in stadial development. Microfilariae ingested by black flies transformed to the L1 larval stage without molting. Successive larval development included molting to the L2 stage and, finally, to the L3 stage, which was infective in humans. The cephalic cap, consisting of a laterally located hook and central stoma, occurs in the first larval stage. The caudal appendix and the laterally located anal opening are apparent in the L1 larva. In the L2 stage, the cephalic cap is lost and the large circular stoma becomes surrounded with elevated flaps. The caudal appendix was lost after larvae molted to the L3 stage, and in its place, 3 terminal papillae developed. Sense organs, such as 2 opposing phasmids and 8 papillae that were arranged into 2 circles, developed in the cephalic region of the L3 larva. The evidence of pathological consequences due to the presence of the L3 larva in the fly host are illustrated and discussed.

Animals↗

Odorant-binding proteins of true bugs. Generic specificity, sexual dimorphism, and association with subsets of chemosensory sensilla.

Odorant-binding proteins (OBPs) in insects occur within olfactory sensilla, and are thought to transport chemical stimuli to receptors on dendrites of sensory neurons. Until recently, knowledge of OBPs in insects was limited to moths and Drosophila. We discovered an antennal-specific protein (Lygus [lineolaris] antennal protein, LAP) with a unique N-terminal sequence in the true bug, Lygus lineolaris. We localized LAP to antennae, determined its molecular weight (16 kDa), and showed that while it was expressed in nymphal antennae, its levels dramatically increased in adults concurrent with increases in numbers of olfactory sensilla and electrical responses to odors. In our current study, we used immunological techniques to demonstrate in more detail that LAP occurs only in antennae, and to show its expression within Lygus species. LAP was expressed more in male antennae than in antennae of females for the Lygus species examined. Anti-LAP did not recognize antennal proteins of two other genera of bugs. Immunocytological studies showed LAP primarily within the sensillar lymph of type 1 and type 4 sensilla on antennae. These observations strongly suggest LAP to be an OBP, and our discovery and characterization of OBPs in true bugs provides a third order for use in the study of evolution of OBPs in insects.

Animals↗

Intergeneric distribution and immunolocalization of a putative odorant-binding protein in true bugs (Hemiptera, Heteroptera).

Lygus antennal protein (LAP) is an olfactory-related protein of the tarnished plant bug Lygus lineolaris (Hemiptera, Heteroptera: Miridae), a hemimetabolous insect. In previous work, a polyclonal antiserum was generated against the N-terminal sequence of LAP; LAP immunoreactivity was strongest in antennae of adult males, but was also present in antennae of adult females and of nymphs. In the current study, LAP immunoreactivity was examined to determine the species specificity and the tissue and cellular localization of LAP expression. Western blot analysis indicated that LAP immunoreactivity was present in the antennae of the male congeners L. lineolaris and L. hesperous, but was not detectable in male antennae of the more distant relatives Podisus maculiventris or Nezara viridula (Hemiptera, Heteroptera: Pentatomidae). Western blot analysis further confirmed that LAP expression was restricted to antennal tissue. Histological analyses showed that LAP expression within the antennae was specifically associated with chemosensory sensilla on the antenna. Within the sensilla, LAP immunoreactivity was distributed throughout the extracellular lumen and was concentrated in dense granules within the cytoplasm of sensillar support cells. LAP immunoreactivity was restricted to a subset of antennal chemosensory sensilla, specifically the multiporous olfactory sensilla. These findings suggest that LAP has an important olfactory function in Lygus sp., possibly related to that of odorant-binding proteins (OBP) found in other insect orders. If so, LAP would be the first OBP-like protein characterized outside the Endopterygota.

Animals↗

Imaging thin and thick sections of biological tissue with the secondary electron detector in a field-emission scanning electron microscope.

A field-emission scanning electron microscope (FESEM) equipped with the standard secondary electron (SE) detector was used to image thin (70-90 nm) and thick (1-3 microns) sections of biological materials that were chemically fixed, dehydrated, and embedded in resin. The preparation procedures, as well as subsequent staining of the sections, were identical to those commonly used to prepare thin sections of biological material for observation with the transmission electron microscope (TEM). The results suggested that the heavy metals, namely, osmium, uranium, and lead, that were used for postfixation and staining of the tissue provided an adequate SE signal that enabled imaging of the cells and organelles present in the sections. The FESEM was also used to image sections of tissues that were selectively stained using cytochemical and immunocytochemical techniques. Furthermore, thick sections could also be imaged in the SE mode. Stereo pairs of thick sections were easily recorded and provided images that approached those normally associated with high-voltage TEM.

Animals↗

Changes in microbial populations on fresh cut spinach.

The microbial populations found on fresh-cut spinach leaves that were stored in gas permeable bags at 10 degrees C for 12 days were examined and identified. The microorganisms consisted of mesophilic aerobic bacteria, psychrotrophic bacteria, Pseudomonadaceae, Enterobacteriaceae, Micrococcaceae, lactic acid bacteria and yeasts. Populations of mesophiles, psychrotrophs, Pseudomonadaceae and Enterobacteriaceae increased sharply during the storage period. The initial populations were 10(7), 10(6), 10(6) and 10(4) CFU.g-1 respectively. Populations reached 10(10) for the mesophiles, psychrotrophs and Pseudomonadaceae and 10(7) CFU.g-1 for Enterobacteriaceae after 12 days of storage. Micrococcaceae, lactic acid bacteria and yeasts remained constant (10(3)-10(4) CFU.g-1. The majority of the bacterial isolates were identified as Pseudomonas fluorescens, Aeromonas caviae and Staphylococcus xylosus. The yeasts, which were most frequently isolated, were classified in the genus Cryptococcus. No pathogens such as Listeria monocytogenes and Salmonella were detected. Observations with low temperature scanning electron microscopy (LTSEM) indicated that the microorganisms were not present on the surface of healthy unbroken leaves. Alternatively, they were found in areas where the cuticle was broken and could be seen infecting the internal palisade parenchyma.

Colony Count, Microbial↗

Characterization of a chicken monoclonal antibody that recognizes the apical complex of Eimeria acervulina sporozoites and partially inhibits sporozoite invasion of CD8+ T lymphocytes in vitro.

A stable chicken hybridoma secreting a monoclonal antibody (mAb) that detects the apical complex of Eimeria acervulina sporozoites has been developed by fusing a thymidine kinase (TK)-deficient chicken myeloma with spleen cells from chickens immunized with sporozoite antigen. The mAb, designated as 6D-12-G10, recognized the apical complex of a sporozoite of 20-21 kDa molecular mass on western blots. Immunoelectron microscopic examination revealed that mAb 6D-12-G10 stained the conoid antigen. Furthermore, mAb 6D-12-G10 inhibited the invasion of sporozoites into CD8+ T cells in vitro. These results suggest that the conoid may play an important role in the recognition and invasion of host cells by Eimeria sporozoites.

Animals↗

Plasmodium vivax: freeze-fracture studies on the ultrastructure of the sporozoites within the salivary gland of the mosquito Anopheles stephensi.

Freeze-fracturing has been used to study the ultrastructure of the sporozoites of the malarial parasite Plasmodium vivax within the salivary gland of the mosquito Anopheles stephensi. The architecture of the pellicular complex of the salivary gland sporozoites was essentially the same as that reported for the intraoocystic forms, but the outline of cross-fractured P. vivax sporozoites was more flattened and crescent shaped as opposed to the circular outline described for the intraoocystic sporozoites. The salivary gland sporozoites of P. vivax also exhibited apical rosettes and a cytosome connected to a food vacuole, two unique structures not previously reported for malarial sporozoites.

Animals↗

Bilateral, perivulval cuticular pores in trichostrongylid nematodes.

A new hypodermal gland was discovered in female nematodes of the family Trichostrongylidae. Because the new structure appears to be associated with the vulva, it was named the perivulval pore. It is similar, based on light and scanning electron microscopy, to phasmids that are located laterally on the tails of nematodes of the class Secernentea. Like phasmids, perivulval pores are paired and bilateral, with cuticular ducts to the surface in the areas of the lateral chords. They are located slightly posterior to the vulva in Haemonchus contortus, Haemonchus placei, Haemonchus similis, Mecistocirrus digitatus, Mazamastrongylus pursglovei, Ostertagia ostertagi, and Cooperia oncophora, but in Trichostrongylus colubriformis they are slightly anterior to the vulva. Because of the location near the vulva and the similarity in structure to phasmids, which are, at least in part, secretory, the perivulval pores should be considered as a possible source of a female attractant for males.

Animals↗

Encapsulation of Streptococcus uberis: influence of storage and cultural conditions.

Streptococcus uberis (n = 100) isolated from bovine mammary secretions were assessed by India ink for expression of capsule. Organisms were evaluated under four conditions; (1) after primary culture on blood agar, (2) following 5 passages on blood agar, (3) after 5 passages in Trypticase Soy Broth (TSB), and (4) after storage in 10% skim milk. Strains from primary culture (44 of 100) were positive for an unstained halo (capsule) by the India ink method. Number of strains expressing capsule decreased greatly after passage and following storage. Freeze-etching followed by electron microscopy confirmed results of India ink preparations. Strains were also cultured in various media to determine influence of medium components on capsule expression. Todd-Hewitt medium supplemented with either serum or egg yolk enhanced the size of capsule expressed. Results of this study may aid researchers investigating the pathogenicity of S. uberis.

Animals↗

Sublateral hypodermal chords in Haemonchus (Nematoda: Trichostrongyloidea): description and potential as a systematic character.

Prominent sublateral hypodermal chords (SHC) are described in Haemonchus spp. Four large SHC are located 1 muscle cell away from the lateral chords from the level of the middle of the esophagus to midbody. The SHC are largest in midbody and merge with the lateral chords at about 65% of the body length from the anterior end. With the aid of transmission electron microscopy, a band of nerve tissue was located parallel and medial to the SHC. Haemonchus spp. and Mecistocirrus digitatus have SHC that are so large they can be identified in thick freehand midbody cross sections cut with a cataract knife. Because SHC have not been described in other Trichostrongylidae, their presence in members of the Haemonchinae may have value as a systematic character.

Animals↗

Pasteuria nishizawae sp. nov., a mycelial and endospore-forming bacterium parasitic on cyst nematodes of genera Heterodera and Globodera.

This study describes Pasteuria nishizawae sp. nov., a fourth species of the genus Pasteuria. This mycelial and endospore-forming bacterium parasitizes the adult females of cyst-forming nematodes in the genera Heterodera and Globodera. The distinct ultrastructural features and unique host range found for this bacterium separate it from two closely related species, Pasteuria penetrans, which parasitizes several species of root-knot nematodes of the genus Meloidogyne, and Pasteuria thornei, which appears to parasitize only one species of the root-lesion nematode, Pratylenchus brachyurus. Because these obligate bacterial parasites of nematodes have not been cultured axenically, the taxonomic relationships described here for each species are based mainly on developmental morphology, fine structure of the respective sporangia and endospores, and their pathogenicity on nematode species.

Animals↗

Distribution of Thylakoid Proteins between Stromal and Granal Lamellae in Spirodela : Dual Location of Photosystem II Components.

We have quantified the lateral distribution of 12 thylakoid proteins of Spirodela oligorrhiza by immunoblot analysis of detergent-derived granal and stromal lamellae. The immunological, ultrastructural, cytochemical, and biophysical measurements each indicated the expected overall separation of photosystem II (PSII) and photosystem I (PSI) components; however, certain proteins were not completely localized to one lamellar fraction. The apoproteins of the light harvesting chlorophyll a/b complex, subunit 1 of PSI and the components of the PSII reaction center (the 32 kilodalton, D2, and cytochrome b(559) proteins) were dually located between granal and stromal lamellae. Proteins associated exclusively with one of the membrane types were: in granal lamellae, the 43 and 51 kilodalton PSII proteins, and in stromal lamellae, the alpha and beta subunits of the proton ATPase.

Journal Article↗

Interactions between spermatozoa and the crypts, cilia, and mucus of the cervix in the ewe.

In ruminants, minor cervical folds, commonly called crypts, temporarily store spermatozoa for a short interval of time between insemination and fertilization. However, the mechanism by which spermatozoa are transported to these crypts and subsequently to the uterus is not known. To study this problem, cervical tissue, which was removed from ewes that were naturally inseminated by rams at estrus, was examined with the scanning electron microscope to determine the physical associations that occur between the spermatozoa and the structural features of the cervix. The study indicates that the spermatozoa generally are not oriented parallel to the longitudinal axis of the cervix, exhibit no consistent association with the cervical cilia, and do not lie in any well defined channels formed by the cervical secretions. Alternatively, the majority of spermatozoa occur as isolated aggregations that lie in or near the shallow folds or crypts of the cervix. The vast numbers of spermatozoa in these aggregations and the lack of any common orientation suggest that some form of external stimulus, such as cervical contractions, might be responsible for the initial mass movement and distribution of spermatozoa in the cervix of the ewe.

Animals↗

Adaptive reorganization of protein and lipid components in chloroplast membranes as associated with herbicide binding.

Cultivation of Spirodela oligorrhiza ( Kurtz ) Hegelm on a sublethal dose of atrazine results in a higher linolenic to linoleic acid ratio in the thylakoid membrane lipids, less starch, more osmiophilic globules, and a reduced stroma lamellar system. Also, the grana become randomly oriented and contain more numerous and elongated lamellae. These alterations in the lipid composition and ultrastructure of the chloroplast resemble those previously observed in triazine-resistant weed biotypes and in chloroplasts developed under low light. Thylakoid membranes from atrazine-adapted plants revealed an additional high-affinity binding constant for [14C]-diuron but the number of diuron binding sites actually decreased by 20 times compared to controls. The 32,000-dalton membrane protein of the chloroplast is synthesized actively, but its breakdown appears decreased compared to control plants. The adaptive reorganization of thylakoid components may be a compensatory mechanism for maintenance of a functional interaction of the proteins and lipids of the photosystem II complex.

Adaptation, Physiological↗

Rhythmical changes in the sensitivity of cotton seedlings to herbicides.

Cotton (Gossypium hirsutum L.) seedlings that were grown under a photoperiod of 12 hours darkness and 12 hours light showed oscillations in their sensitivity to the herbicides sodium 5-(2-chloro-4-trifluoromethyl)-phenoxy)-2-nitrobenzoate (acifluorfen), butyl 2-(4-((5-(trifluoromethyl)-2-pyridinyl)oxy)phenoxy)propanoate (fluazifop) and 3-isopropyl-1H-2,1,3-benzothiadiazin-4(3H)-one 2,2-dioxide (bentazon). Sensitivity was expressed in appearance of necrotic areas on the cotyledons and in decreased growth of the shoot. The seedlings were least sensitive in the beginning and middle of the light period, then the sensitivity increased and reached its maximum during the beginning and middle of the dark period and then declined. Seedlings grown from germination under continuous light exhibited very small or no oscillations in sensitivity. The oscillations in sensitivity were entrained by one cycle of darkness and light. A cycle of 12 hours darkness and 12 hours light triggered the greatest oscillations while either increasing or decreasing the duration of the dark period resulted in smaller oscillations. Apparently, these oscillations in sensitivity to herbicides were endogenously controlled since after entrainment they continued irrespective of the light conditions.

Journal Article↗

Cyclic changes in the surface structure of the cervix from the ewe as revealed by scanning electron microscopy.

Thirty parous ewes were divided into six groups and sacrificed on day 0 (first day of estrus), 1, 2, 10, 15 or 16 of the estrous cycle. The cervices were removed immediately and processed for examination with the scanning electron microscope. Observation of the tissues reveals that the surface of the cervix is highly convoluted, which results in the formation of numerous folds or crypts. Two forms of columnar epithelial cells, a ciliated and a non-ciliated cell with microvilli, line the luminal surface of the cerix in the day 10, luteal-phase ewes. However, on day 15, 2 days before estrus, the non-ciliated cells differentiate into two morphologically distinct types of secretory cells. One type forms when the apex of the non-ciliated cell dilates outward into the lumen of the cervix. Concurrent with apical enlargement, the microvilli are lost and the limiting cell membrane becomes smooth. The other type of cell is characterized by only a slight apical swelling. Consequently, remnants of microvilli along with secretory granules can be observed on the limiting membrane of this cell. Both cells release a particulate component, which is believed to be a precursor of mucus, into the lumen of the cerix. These particles undergo a series of morphological transformations to form a fibrillar layer, generally referred to as 'cervical mucus', that covers the epithelial surface at estrus. One to 2 days following the onset of estrus, the fibers become more closely assoicated with amorphous material that begins to coagulate, thereby revealing the underlying ciliated and non-ciliated cells that characterize the cervix of the luteal-phage ewe. The cyclical variation in secretory cells and factors that may influence that structural transformations which occur in mucus are discussed.

Animals↗