Electrophoretic comparison of the genomes of North American bluetongue viruses, one Australian bluetongue virus, and three other related orbiviruses

The genomes of U.S. bluetongue viruses, an Australian bluetongue virus, and three other related orbiviruses were analyzed by polyacrylamide gel electrophoresis. The genomes were comprised of ten segments of double-stranded (ds) RNA. Estimates of the molecular weights of the dsRNA segments revealed that the U.S. bluetongue serotypes were remarkably similar. Although the dsRNA profiles of the viruses exhibited common segments, each virus had a distinct dsRNA profile. The usefulness of the genome analysis as a diagnostic tool for identification and for epidemiologic studies is discussed.     

The recreational and commercial importance of feral swine in Florida: Relevance to the possible introduction of African swine fever into the U.S.A.

Florida has the largest population of feral swine in the U.S.A. Feral swine have recreational and economic importance to hunters, trappers, taxidermists, and also to dealers who sell feral swine to hunting clubs. Hunters spent over 522 00 man-days hunting feral swine during the 1980–1981 hunting year. They killed nearly 103 000 hogs which they valued at over $58 each, for a total value of nearly $6 million. Trappers caught 25 000 head valued at about $ 700 000, and landowners collected about $1.2 million in hunting lease fees. Taxidermists received about $389 000 in gross income from mounting feral swine. Sales of feral swine to hunting clubs amounted to about $81 000.

Sales of feral swine through commercial livestock auctions were limited to 1620 head valued at only $16 800 in 1980. Thus, feral swine are important to Florida from a recreational standpoint, but not from the perspective of commercial agriculture.

If African swine fever were to enter Florida, there are many socio-economic factors associated with feral swine that must be carefully evaluated for eradication to be successful.     

Failure to detect luteinizing hormone-releasing hormone-like bioactivity in fluid of preovulatory ovine follicles

To determine if a luteinizing hormone-releasing hormone (LH-RH)-like substance accumulates in fluid of follicles destined to ovulate, we obtained the fluid component of preovulatory ovine follicles at various times before, during, and after the endogenous surge in pituitary secretion of LH. The fraction of follicular fluid suspected of containing LH-RH was isolated by reverse-phase column chromatography. Luteinizing hormone releasing activity of the preparation was ascertained using an anterior pituitary cell bioassay of high sensitivity. In no instance was activity of a peptide similar to LH-RH revealed in fluid obtained from preovulatory follicles of ewes.     

Pasteurella haemolytica: purification of saline-extractable proteins by isoelectrofocusing

A procedure was developed for separating antigens associated with a saline extract of Pasteurella haemolytica serotype 1. Seven antigens were identified by immunoelectrophoresis to be associated with the extract. The extract was subjected to preparative isoelectrofocusing in a pH range of 3-10. The majority of extracted proteins were found to have pI's of 4-6, whereas the carbohydrate antigen(s) were distributed over a pI range of 3.0-8.0. The fractions that were of interest were pooled and refocused in a narrower pH range to improve resolution of the protein antigens. Specific antigens from defined pH ranges were pooled to form 6 antigen groups. These antigen groups were examined further by immunoelectrophoresis, analytical isoelectrofocusing, and sodium dodecyl sulphate-polyacrylamide gel electrophoresis. The molecular weights of the proteins found in the capsular extracts ranged from 33 k to greater than 80 k. Injection of mice with capsular extract or antigen Groups 1-6 in Freund's incomplete adjuvant resulted in a serum antibody response to the various antigens as detected by an enzyme-linked immunosorbent assay. Significant protection (P less than 0.05) against challenge with virulent P. haemolytica was seen in mice injected with antigen Groups 2 and 4. Six calves were immunized with saline extract. These calves had greater resistance to experimental pneumonic pasteurellosis than did 6 non-vaccinated calves. A serum antibody response to the crude extract and to each antigen group was detected in vaccinated calves by an enzyme-linked immunosorbant assay.     

Isolation of Pasteurella haemolytica and correlation with serum antibody response in clinically normal beef calves

Bacteria from the nasal cavity and trachea were cultured, and serum antibody titers determined for Pasteurella haemolytica serotype 1 in 164 beef calves obtained from a closed herd on range pasture. At the first sampling, P. haemolytica serotype 1 was cultured from 16.4% of the calves. Antibody titers were determined by a quantitative fluorimetric method and the mean titer was 9.5 +/- 5.8. Fifty-seven randomly selected calves were used to study the correlation of serum antibody response and positive culture of P. haemolytica under natural conditions. Clinical signs of respiratory disease were not observed in those calves. During the observation periods, there was a two-fold increase in the percentage of calves that were culture positive. There was no significant difference between mean serum antibody titers or frequency distribution of antibody titers from the two samplings. Comparisons between serum antibody titers, rise in titers, and P. haemolytica isolation failed to reveal any significant correlation. Of the 9 calves that had a decline in antibody titer to P. haemolytica, none was culture positive. Seroconversion to respiratory viruses did not correlate with P. haemolytica related variables.     

The role of specific immunoglobulins in antibody-dependent cell-mediated cytotoxicity assays during Babesia bovis infection

A stabilate prepared from Babesia bovis-infected Boophilus microplus ticks was used to infect intact adult cattle. Whole sera and immunoglobulin fractions from representative sera were tested by complement fixation (CF), indirect fluorescent antibody (IFA), and antibody-dependent cell-mediated cytotoxicity (ADCC) assays. The last test utilized 51Cr-labeled chicken erythrocytes coated with Babesia bovis antigen as targets. Mononuclear cell preparations, obtained from peripheral blood of normal donors and consisting of lymphocytes with 2--6% large monocytes, were used as the source of effector cells. Antibody activity was detected by all tests between 14 and 16 days following infection. Specific IgM and IgG1 were reactive in both CF and IFA tests, although the development of high titers was attributable to IgG, alone. The ADCC activity was restricted to IgG1 fractions and was greater in those sera or fractions with greater CF activity. No activity was demonstrated in IgG2 fractions by any test used.     

Landowner Perceptions of Legal Liability for Using Prescribed Fire in the Southern Plains,United States

Suppression of fire in the Southern Plains has led to proliferation of woody plants and fuel load accumulation that spurs wildfires. These effects have led to calls for widespread application of prescribed fire to reduce fuel loads, but there is substantial landowner resistance to the use of this land management tool. Here we explore factors that affect perceptions of landowners in the Southern Plains about prescribed fire liability and their willingness to apply this land management tool. This region was selected for the study because of the preponderance of private landholdings and widespread woody plant encroachment. The study used a mail survey of 1 853 landowners in 16 counties in Texas and Oklahoma, resulting in a data set from 680 respondents (37% useable response rate). Logistic regression models were developed to test three hypotheses relating to the likelihood that a landowner will apply prescribed fire. The study corroborated that landowners who perceived higher levels of fire-related legal liability were less likely to apply prescribed fire on their land or assist with its application on other properties. In addition, burn bans were found to inhibit landowner willingness to apply fire during periods that result in higher woody plant mortality. Oklahoma respondents, landowners who believed prescribed fire to be an affordable woody plant management tool, and members of prescribed burning associations (PBAs) were more likely to use prescribed fire. These results have important implications for policies aimed at overcoming resistance to the use of prescribed fire to curb woody plant encroachment and reduce fuel load accumulation. Specifically, language in state statutes pertaining to prescribed fire should be modified to reduce landowner concerns over legal liability; PBAs should be established more widely; and public cost-sharing funds for woody plant management should prioritize prescribed fire.     

Ovarian inhibition of pulsatile luteinizing hormone secretion in prepuberal holstein heifers

Fifteen prepuberal Holstein heifers were utilized to examine pulsatile luteinizing hormone (LH) secretion before and after ovariectomy. Heifers were ovariectornized at 3, 6 or 9 months of age (n=5/group) and scheduled for blood sampling at 1 week before, 1 week after and 4 weeks following ovariectomy. During each 8 hr sampling period (0600–1400 hr), blood samples (10 ml) were collected via indwelling jugular canulae at 10 min intervals. Prior to ovariectomy, mean plasma LH concentration and both number and amplitude of LH pulses per 8 hr sampling period were similar (P>.05) among age groups, and the absence of a pulsatile LH secretion profile was accompanied by a low mean LH concentration. Within 1 week after ovariectomy, both number of LH pulses and mean LH concentrations increased (P<.O1) in all age groups. Between 1 and 4 weeks after ovariectomy, both amplitude of LH pulses and mean LH concentrations increased (P<.O1) when the data from the three age groups were combined. We conclude that ovarian inhibition of pulsatile LH secretion is established by 3 months of age and is maintained through 9 months of age. In addition, the initial elevation mean plasma LH concentration is due to greater pulse frequency, while the subsequent rise in mean LH concentration reflects increased amplitude of LH pulses.     

Potential Spread of Cheatgrass and Other Invasive Species by Feral Horses in Western Colorado

The invasive grass cheatgrass (Bromus tectorum L.) presents major challenges for land management and habitat conservation in the western United States. Feral horses (Equus ferus caballus) have become overabundant in some areas of the West and can impact fragile semiarid ecosystems. Amid ongoing efforts to control cheatgrass in the Great Basin, we conducted a study to determine if feral horses contribute to the spread of cheatgrass through distribution via their feces. We collected feral horse fecal samples from Little Book Cliffs Herd Management Area in western Colorado in 2014. Fecal samples were dried, and 20 from each of 3 collection sessions were cultivated to examine germination success. Six species germinated from 18 samples (30%; mostly one plant per sample where germination occurred), including cheatgrass from 8% of samples. In a separate study we examined the diet of this same horse population using fecal plant DNA barcoding. Plant species that germinated were rare in the diet and germinated from fewer samples than expected relative to their detection in the diet. Our results suggest that feral horses could be contributing to cheatgrass propagation. Native ungulates and domestic cattle also have this potential. Although management of all large ungulates is necessary to mitigate cheatgrass spread, control of feral horse numbers is particularly necessary.     

Cellular composition of granulomatous lesions in gut-associated lymphoid tissues of goats during the first year after experimental infection with Mycobacterium avium subsp. paratuberculosis

Mycobacterium avium subsp. paratuberculosis (MAP) causes lesions in naturally and experimentally infected ruminants which greatly differ in severity, cellular composition and number of mycobacteria. Morphologically distinct lesions are already found during the clinically inapparent phase of infection. The complex local host response and number of MAP were characterized at the initial sites of lesions, organized gut-associated lymphoid tissue, in experimentally infected goats. Tissues were collected at 3, 6, 9 and 12 month post-inoculation (mpi) from goat kids that had orally received 10 times 10 mg of bacterial wet mass of MAP (JII-1961). The cellular composition of lesions in Peyer's patches in the jejunum and next to the ileocecal valve was evaluated in 21 MAP-inoculated goats, where lesions were compared with unaltered tissue of six control goats. CD68⁺, CD4⁺, CD8⁺, γδ T lymphocytes, B lymphocytes and plasma cells, MHC class II⁺ and CD25⁺ cells were demonstrated by immunohistochemistry in serial cryostat sections.At 3 mpi, extensive granulomatous infiltrates predominated, consisting of numerous epitheloid cells admixed with many CD4 and γδ T lymphocytes. Only single MAP were detected. This indicates a strong cellular immune reaction able to control MAP infection. γδ T lymphocytes were markedly increased in this type of lesion which may reflect their important role early in the pathogenesis of paratuberculosis. At 9 and 12 mpi, divergent lesions were observed which may reflect different outcomes of host–pathogen interactions. In five goats, minimal granulomatous lesions were surrounded by extensive lymphoplasmacytic infiltrates and no MAP were detected by immunohistochemistry. This was interpreted as effective host response that was able to eliminate MAP locally. In three goats, decreased numbers of lymphocytes, but extensive granulomatous infiltrates with numerous epitheloid cells containing increased numbers of mycobacteria were seen. This shift of the immune response resulted in uncontrolled mycobacterial multiplication. Focal and multifocal circumscribed granulomatous infiltrates of mainly epitheloid cells may represent sites of new infection, since they were observed in goats at all times after inoculation. Their presence in goats with minimal granulomatous lesions surrounded by extensive lymphoplasmacytic infiltrates may indicate that despite the local clearance, the infection may be perpetuated.The complex cellular immune reactions postulated for the pathogenesis of paratuberculosis were demonstrated at the local sites of infection. These early host–pathogen interactions are most likely essential for the eventual outcome of the MAP infection.