GAPEWORM (SYNGAMUS SPP.) PREVALENCE IN WISCONSIN GREATER PRAIRIE CHICKENS (TYMPANUCHUS CUPIDO PINNATUS).

Under Wisconsin state law, the greater prairie chicken (GRPC; Tympanuchus cupido pinnatus) has been listed as a threatened species since 1976. In 2014-15, we conducted a pilot study to determine the prevalence and intensity of gapeworms (Syngamus spp.) in female Wisconsin GRPCs collected from 2 monitored populations. We captured 62 female GRPCs using walk-in-style traps for females and night lighting for juveniles ≥45 days of age. From these individuals, we collected 15 carcasses of radio-marked birds, most of whom died due to predation events. Through dissection, we identified gapeworm in 20% of examined carcasses and report an intensity ranging between 4 and 74 worms.

Metabolic Flexibility in Response to Within-Season Temperature Variability in House Sparrows.

The climatic variability hypothesis (CVH) posits that more flexible phenotypes should provide a fitness advantage for organisms experiencing more variable climates. While typically applied across geographically separated populations, whether this principle applies across seasons or other conditions (e.g., open vs. sheltered habitats) which differ in climatic variability remains essentially unstudied. In north-temperate climates, climatic variability in winter usually exceeds that in summer, so extending the CVH to within-population seasonal variation predicts that winter phenotypes should be more flexible than summer phenotypes. We tested this prediction of the within-season extension of the CVH by acclimating summer and winter-collected house sparrows (Passer domesticus) to 24, 5, and -10°C and measuring basal metabolic rate (BMR) and summit metabolic rate (Msum = maximum cold-induced metabolic rate) before and after acclimation (Accl). To examine mechanistic bases for metabolic variation, we measured flight muscle and heart masses and citrate synthase and ß-hydroxyacyl coA-dehydrogenase activities. BMR and Msum were higher for cold-acclimated than for warm-acclimated birds, and BMR was higher in winter than in summer birds. Contrary to our hypothesis of greater responses to cold Accl in winter birds, metabolic rates generally decreased over the Accl period for winter birds at all temperatures but increased at cold temperatures for summer birds. Flight muscle and heart masses were not significantly correlated with season or Accl treatment, except for supracoracoideus mass, which was lower at -10°C in winter, but flight muscle and heart masses were positively correlated with BMR and flight muscle mass was positively correlated with Msum. Catabolic enzyme activities were not clearly related to metabolic variation. Thus, our data suggest that predictions of the CVH may not be relevant when extended to seasonal temperature variability at the within-population scale. Indeed, these data suggest that metabolic rates are more prominently upregulated in summer than in winter in response to cold. Metabolic rates tended to decrease during Accl at all temperatures in winter, suggesting that initial metabolic rates at capture (higher in winter) influence metabolic Accl for captive birds.

The Rous sarcoma virus Env glycoprotein contains a highly conserved motif homologous to tyrosine-based endocytosis signals and displays an unusual internalization phenotype.

The cytoplasmic domains of retroviral transmembrane (TM) glycoproteins contain conserved sequence motifs that resemble tyrosine-based (YXXO-type) endocytosis signals. We have previously described a mutant Rous sarcoma virus (RSV) Env protein, Env-mu26, with an L165R mutation in the membrane-spanning domain (MSD) of TM, that exhibited dramatically decreased steady-state surface expression (G. L. Davis and E. Hunter, J. Cell Biol. 105:1191-1203, 1987; P. B. Johnston, J. Y. Dong, and E. Hunter, Virology 206:353-361, 1995). We now demonstrate that the tyrosine of the Y(190)RKM motif in the RSV TM cytoplasmic domain is crucial for the mu26 phenotype and is part of an efficient internalization signal in the context of a mutant MSD. In contrast, despite the presence of the Y(190)RKM motif, wild-type RSV Env is constitutively internalized at a slow rate (1.1%/min) more characteristic of bulk uptake during membrane turnover than of active clustering into endocytic vesicles. The mu26 mutation and two MSD mutations that abrogate palmitoylation of TM resulted in enhanced Env endocytosis indicative of active concentration into coated pits. Surprisingly, an Env-Y190A mutant was apparently excluded from coated pits since its uptake rate of 0.3%/min was significantly below that expected for the bulk rate. We suggest that in RSV Env an inherently functional endocytosis motif is silenced by a counteracting determinant in the MSD that acts to prevent clustering of Env into endocytic vesicles. Mutations in either the cytoplasmic tail or the MSD that inactivate one of the two counteracting signals would thus render the remaining determinant dominant.

Bacteria and nematodes in the conjunctiva of mule deer from Wyoming and Utah.

Swabs of conjunctiva were collected from 44 live and 226 hunter-harvested mule deer (Odocoileus hemionus) from Wyoming and Utah (USA). We identified 29 gram negative and 22 gram positive bacterial taxonomic categories, but many isolates from hunter-harvested animals were environmental contaminants. Staphylococcus spp. and Micrococcus spp. were the most common gram positive bacteria isolated, and Enterobacter spp., Escherichia coli, and Pseudomonas spp. were common gram negative bacteria isolated. Thelazia californiensis were found in 15% of hunter-harvested deer in Utah in 1994 and in 8% in 1995. Nematodes were found in 40% of live deer in 1995 and 66% in 1996. Three live animals showed clinical signs of infectious keratoconjunctivitis (IKC) in 1996, but pathogenic bacteria were not isolated from these individuals. Hemolytic, non-piliated Moraxella ovis was isolated from two clinically normal live deer in 1996 and isolates were similar to those cultured from IKC cases from Wyoming and Utah.

Association of Moraxella ovis with keratoconjunctivitis in mule deer and moose in Wyoming.

Six cases of infectious keratoconjunctivitis (IKC) in mule deer (Odocoileus hemionus) and moose (Alces alces) in Wyoming (USA) were investigated during fall and winter of 1995 and 1996. Excessive lacrimation, mucopurulent conjunctivitis, keratitis, and corneal opacity were observed in mule deer. Moose had severe mucopurulent conjunctivitis, keratitis, and corneal ulceration. Hemolytic, non-piliated Moraxella ovis was isolated from two mule deer and two moose. We attempted to reproduce IKC in three mule deer fawns using an isolate of M. ovis from a clinically affected mule deer. These fawns did not develop clinical signs of infection and the bacterium was not reisolated from inoculated deer. Inoculated deer may not have developed clinical signs because deer were not exposed to ultraviolet light or mechanical insult before inoculation. In addition, the isolate used for inoculation may have lost virulence factors through passage, or M. ovis may not have been the primary pathogen responsible for clinical disease in the natural cases of IKC we investigated. The etiology of IKC in free-ranging wild ruminants remains poorly understood.

Lack of relay toxicity in ferret hybrids fed carbaryl-treated prairie dogs.

Carbaryl (1-napthol methylcarbamate) is being considered for control of fleas on prairie dogs (Cynomys spp.) used in black-footed ferret (Mustela nigripes) recovery in the western United States. The potential for relay toxicity in ferrets was determined by feeding carbaryl treated prairie dogs to black-footed ferret x Siberian polecat (M. eversmanni) hybrids. Adult prairie dogs were treated topically with 2.5 g of commercial 5% carbaryl dust sold as flea powder. After 14 days prairie dogs were killed and fed to ferrets. Potential for relay toxicity was evaluated by analyzing ferret blood cholinesterase (CHe), prairie dog brain Che, and hepatic carbamate concentration. There was no difference between pre- and post-exposure blood CHe activity, nor did treated prairie dog brain CHe differ significantly from controls. Post-exposure blood CHe did not exhibit reactivation after dilution in aqueous buffer. Hepatic carbaryl concentrations were less than detection limits (50 ppb). Based on these results, we conclude that short-term use of carbaryl for flea control on prairie dogs does not pose a hazard of relay toxicity in black-footed ferrets.

Mutations within the putative membrane-spanning domain of the simian immunodeficiency virus transmembrane glycoprotein define the minimal requirements for fusion, incorporation, and infectivity.

The membrane-spanning domain (MSD) of a number of retroviral transmembrane (TM) glycoproteins, including those from the human and simian immunodeficiency viruses (HIV and SIV), have been predicted to contain a charged arginine residue. The wild-type SIV TM glycoprotein is 354 amino acids long. The entire putative cytoplasmic domain of SIV (amino acids 193 to 354) is dispensable for virus replication in vitro, and such truncation-containing viruses are capable of reaching wild-type titers after a short delay. We show here that further truncation of eight additional amino acids to TM185 results in a protein that lacks fusogenicity but is, nevertheless, efficiently incorporated into budding virions. By analyzing a series of nonsense mutations between amino acids 193 and 185 in Env expression vectors and in the SIVmac239 proviral clone, a region of the SIV TM that contains the minimum requirement for glycoprotein-mediated cell-to-cell fusion and that for virus replication was identified. Virus entry and infectivity were evident in truncations to a minimum of 189 amino acids, whereas cell-cell fusion was observed for a protein of only 187 amino acids. Glycoprotein was efficiently incorporated into budding virions in truncations up to 185 amino acids, indicating that such proteins are membrane anchored and are transported to the cell surface. However, truncation of the TM to 180 amino acids resulted in a protein that displays a transport defect and may be retained in the endoplasmic reticulum. Based on our analyses of these mutants, an alternative model for the MSD of SIV is proposed. Our model suggests that membrane-imbedded charged residues can be neutralized by side-chain interactions with lipid polar head groups. As a consequence, the membrane-spanning region can be reduced by more than a helical turn. This new model accounts for the ability of truncations within the predicted MSD to remain membrane anchored and maintain biological activity.

Analysis of the cleavage site of the human immunodeficiency virus type 1 glycoprotein: requirement of precursor cleavage for glycoprotein incorporation.

Endoproteolytic cleavage of the glycoprotein precursor to the mature SU and TM proteins is an essential step in the maturation of retroviral glycoproteins. Cleavage of the precursor polyprotein occurs at a conserved, basic tetrapeptide sequence and is carried out by a cellular protease. The glycoprotein of the human immunodeficiency virus type 1 contains two potential cleavage sequences immediately preceding the N terminus of the TM protein. To determine the functional significance of these two potential cleavage sites, a series of mutations has been constructed in each site individually, as well as in combinations that altered both sites simultaneously. A majority of the mutations in either potential cleavage site continued to allow efficient cleavage when present alone but abrogated cleavage of the precursor when combined. Despite being transported efficiently to the cell surface, these cleavage-defective glycoproteins were unable to initiate cell-cell fusion and viruses containing them were not infectious. Viruses that contained glycoproteins with a single mutation, and that retained the ability to be processed, were capable of mediating a productive infection, although infectivity was impaired in several of these mutants. Protein analyses indicated that uncleaved glycoprotein precursors were inefficiently incorporated into virions, suggesting that cleavage of the glycoprotein may be a prerequisite to incorporation into virions. The substitution of a glutamic acid residue for a highly conserved lysine residue in the primary cleavage site (residue 510) had no effect on glycoprotein cleavage or function, even though it removed the only dibasic amino acid pair in this site. Peptide sequencing of the N terminus of gp41 produced from this mutant glycoprotein demonstrated that cleavage continued to take place at this site. These results, demonstrating that normal cleavage of the human immunodeficiency virus type 1 glycoprotein can occur when no dibasic sequence is present at the cleavage site, raise questions about the specificity of the cellular protease that mediates this cleavage and suggest that cleavage of the glycoprotein is required for efficient incorporation of the glycoprotein into virions.

Knowledge, awareness and attitudes of guardians in the community towards distribution of the HPV vaccine to primary and secondary school children in Trinidad

OBJECTIVE: To investigate the knowledge, awareness and attitudes of guardians towards the distribution of Gardasil to school children in the North Central Regional Health Authority jurisdiction of Trinidad. DESIGN AND METHODS: A cross sectional study utilizing a questionnaire was employed to collect data from 368 participants who reside in the North Central division of Trinidad. The statistical analysis included chi square tests and logistical regression. RESULTS: There was an increase in the approval rate of Human Papillomavirus (HPV) vaccine (32.4%) after the participants received further education. Guardians preferred the 12-14 (24.2%) and 9-11 (23.1%) age groups for vaccination. Religion did not influence the decision to vaccinate. About half (49.2%) of parents believed vaccination would not cause promiscuity. Women were more likely to approve of the vaccine (p=0.023). Most participants did not talk to a doctor about the vaccine (70.7%). The general opinion (44.9%) was that insufficient information was imparted for an informed decision to be made. CONCLUSION: There was approval of the HPV vaccine despite risks and lack of knowledge. A more effective vaccination program requires prolonged sensitization of the public on the HPV vaccine.