Identification of resistance to Aspergillus flavus infection in cotton germplasm

Natural resistance in cottonseed to Aspergillus flavus infection has not been explored to date. A green fluorescent protein (GFP) expressing A. flavus strain was used to assess the resistance of seed from 35 cotton varieties from Gossypium arboreum, G. barbadense, and G. hirsutum. Mature cotyledons devoid of seed coat were wounded, inoculated, and assessed for innate resistance to A. flavus infection. Of the initial 35 varieties tested, we observed a range of resistance to infection in representatives of all three species. A subset of 15 representatives was further analyzed. Within this group, G. arboreum cultivar A2 186 and G. hirsutum cultivar SA 1582 were most resistant to fungal infection. The most susceptible cultivar in this group was G. hirsutum SA 1595. The remaining 12 representatives tested in the secondary screen (3 G. arboreum, 3 G. barbadense, and 6 G. hirsutum lines) exhibited intermediate resistance. We did not observe any relationship between species and resistance. Within each species, there was a range of responses to fungal infection. Future studies using this methodology to screen additional diploid and tetraploid cotton lines may enable us to identify naturally resistant germplasm that can be used to develop cotton with enhanced resistance to fungal infection.     

Effects of porcine relaxin on induced parturition in beef heifers

Two-year-old crossbred beef heifers were used to test the effects of porcine relaxin (pRelaxin) alone, or in combination with dexamethasone, on the induction of parturition, the incidence of dystocia, and retained placentas. Effects of treatment on pelvic area, postpartum interval, milk production, colostrum quality, calf birth weight, calf vigor, and calf performance were also evaluated. On Day 275 of gestation, heifers from two fetal-sire groups were randomly assigned to one of four groups in a 2 × 2 factorial design and received: no treatment (controls, N = 19), 20 mg of dexamethasone intramuscularly (im) (n = 22), 5 mg of pRelaxin (3,000 U/mg) im (n = 19), or 20 mg of dexamethasone plus 5 mg of pRelaxin (n = 17). Length of gestation (in days) was less (P < 0.05) in heifers treated with dexamethasone (279.8 ± 1.0) than in controls (286.6 ± 0.9), but was not influenced (P > 0.05) by treatment with pRelaxin. The incidence of retained placentas in heifers treated only with dexamethasone (27.3%) was not reduced by concomitant treatment with pRelaxin (35.3%). Retained placentas were not observed in any control heifers and in only one heifer (5.2%) treated solely with pRelaxin. Ease of calving (1 = unassisted, 5 = abnormal presentation) was not influenced by treatment (P > 0.05), even though birth weights (in kilograms) of calves from heifers treated with dexamethasone (36.4 ± 0.8) were less (P < .01) than those of calves from nondexamethasone-treated heifers (39.2 ± 0.8). Dexamethasone tended to reduce (P < 0.07) calf vigor (1 = healthy and strong, 5 = dead on arrival; 1.48 ± 0.11 vs. 1.18 ± 0.11), but was not (P > 0.05) influenced by pRelaxin. The duration of the postpartum anestrous interval (73.1 ± 1.8 d across groups) and pelvic areas following treatment and parturition were not influenced (P > 0.05) by dexamethasone or pRelaxin. Although determinants of colostrum quality (P < 0.01) and quantity (P < 0.08) of milk produced were influenced by dexamethasone, adjusted 205-d weights of calves did not differ (P > 0.05) among groups. In conclusion, treatment with pRelaxin alone failed to induce parturition or, when combined with dexamethasone, to reduce the incidence of retained placentas.     

Mechanisms responsible for high levels of permethrin resistance in the house fly

The mechanisms responsible for > 6000-fold permethrin resistance in a pyrethroid-selected strain of house fly, Learn-PyR, were investigated. Through electrophysiological, in vitro metabolism, in vivo penetration and synergism studies it was demonstrated that the resistance mechanisms consisted of enhanced metabolic detoxification via the mixed-function oxidase (MFO) system, target-site insensitivity and decreased cuticular penetration. The major resistance mechanism was the MFO-mediated detoxification. The elevated MFO activity was correlated with higher levels of cytochrome P-450, cytochrome b₅ and NADPH-cytochrome c reductase activity. The kinetics of the latter showed similar K_m but greater V_max values in the Learn-PyR than in the susceptible strain, suggesting that the elevated activity was due to an altered amount, but not an altered form, of the enzyme. The Learn-PyR strain showed widely varying levels of resistance to the pyrethroids tested. Comparison of the pyrethroid structures with the resistance ratios revealed that resistance was highest in the presence of an unsubstituted phenoxybenzyl alcohol moiety. Substitution or certain modifications of the alcohol moiety reduced the level of resistance. Structure of the acid moiety or the presence or absence of an a-CN group did not affect the resistance level. These results are discussed with reference to the resistance mechanisms present.     

Activity of two novel insecticides against permethrin-resistant Pseudoplusia includens

Two experimental insecticides, AC 303,630 and MK 244, were tested against a laboratory colony and three field strains of Pseudoplusia includens (Walker). Topical application bioassays indicated that permethrin resistance in the field strains ranged from 3.9 to 489.0-fold. In leaf dip bioassays, LC₅₀ and LC₉₀ values for AC 303,630 ranged from 6.7 to 15.1 mg litre ?1 and 8.7 to 28.2 mg litre ?1, respectively, for third-instar larvae. The Louisiana 1991 field strain was significantly more susceptible to AC 303,630 than the laboratory and other field strains. The LC₅₀ (but not LC₉₀) for the Louisiana 1992 field strain was significantly higher than that of the laboratory strain. However, there was no difference in toxicity of AC 303,630 between the field strain with the highest level of permethrin resistance and the laboratory strain. LC₅₀ and LC₉₀ values for MK 244 in leaf dip bioassays ranged from 0.014 to 0.023 mg litre ?1 and 0.079 to 0.174 mg litre ?1, respectively. There were no significant differences in LC 50 or LC 90 among any of the strains for MK 244. Field trials in soybean were also conducted in 1991 and 1992 in an area of Louisiana where permethrin efficacy against P. includens has declined. In both years, AC 303,630 at 0.11–0.22 kg ha ?1, and MK 244 at 0.0042–0.0084 kg ha ?1, provided significantly better control than permethrin at 0.11 kg ha ?1, and control equal to the recommended standard, thiodicarb. These studies indicate no cross-resistance exists between the experimental insecticides and permethrin.     

In vitro pharmacologic effect of two endothelin-1 antagonists on equine colonic arteries and veins

OBJECTIVE: To evaluate the effectiveness of 2 potential endothelin (ET)-1 antagonists in blocking the contractile responses of equine colonic vessels to increasing concentrations of ET-1. SAMPLE POPULATION: Mesenteric vessels from 6 clinically healthy horses. PROCEDURE: Colonic vessels (arterial and venous rings) were placed in organ baths with oxygenated Tyrode solution at 37 C. Each was attached to a force transducer interfaced with a polygraph, and 2 g of tension was applied and equilibrated for 45 minutes. Then, B-1 (PD 142893) and B-2 (PD 145065) ET-1 antagonists were tested. One ring from each vessel type was used as a control for determining concentration-response relationships of ET-1 (10(-10) to 10(-6)M). Three rings of each vessel type were incubated with 3 concentrations of each antagonist (10(-7), 10(-6), and 10(-5) M) for 30 minutes before ET induced contractions were determined. The maximum contractile response and pA2 values were determined. RESULTS: Vessels contracted in a concentration-dependent manner to ET-1. Arteries responded slowly but reached greater contractions. Veins responded immediately with sustained contractions. Both antagonists inhibited contractions in a concentration-dependent manner with significant differences at 10(-6) and 10(-5)M for arteries and 10(-5) M for veins. Complete blockade of contractions was observed with B-2 (10(-5)M). The pA2 values for B-1 were 8.26 and 6.82 for arteries and veins, respectively, whereas they were 8.25 and 7.21 for B-2. CONCLUSION AND CLINICAL RELEVANCE: Both antagonists effectively blocked ET-1-induced contractions of equine colonic vessels. Because B-2 is water soluble and caused complete blockade at 10(-5) M, it appears to be the preferred antagonist.     

Expression of interleukin-1beta in the digital laminae of horses in the prodromal stage of experimentally induced laminitis

OBJECTIVE: To study expression of interleukin-1beta (IL-1beta) in the digital laminae of horses in the prodromal stage of experimentally induced laminitis. ANIMALS: 8 healthy adult horses with no signs of laminitis. PROCEDURES: Black walnut extract was administered via nasogastric tube to 4 horses, and water was administered to the remaining 4 (controls). Complete blood counts and physical examinations were performed every 30 minutes after administration of black walnut extract or water. General anesthesia was induced when total WBC count decreased by 30% in horses given the black walnut extract and 3 hours after water administration in control horses. The left forefoot was perfusion fixed with neutral-buffered 10% formalin, and paraffin-embedded sections of the digit were used for in situ hybridization with an equine-specific IL-1beta probe. RESULTS: IL-1beta mRNA expression was observed in perivascular cells of the small laminar venules and capillaries in all 4 horses given black walnut extract and in interstitial cells remote from the microvasculature in 1 of the 4. Other cellular components of the laminar tissue and cellular components of the digital arterioles and veins did not exhibit IL-1beta mRNA expression. Expression of IL-1beta mRNA was not detected in laminae from control horses. CONCLUSIONS AND CLINICAL RELEVANCE: Results suggest that IL-1beta mRNA is expressed by perivascular cells in the laminar tissues of horses in the prodromal stage of experimentally induced laminitis. This provides evidence of an inflammatory process during the prodromal stage of laminitis, indicating that local digital proinflammatory cytokine expression may be an initiating factor in laminitis.     

Causes of death or reasons for euthanasia in military working dogs: 927 cases (1993-1996)

OBJECTIVE: To determine causes of death or reasons for euthanasia in a population of military working dogs. DESIGN: Retrospective study. ANIMALS: 927 military working dogs. PROCEDURE: Records of all military working dogs that died during the period from 1993 to 1996 were evaluated for cause of death or reason for euthanasia by review of necropsy and histopathology reports, death certificates, and daily clinical treatment sheets. A single primary cause of death or euthanasia was determined. RESULTS: Although sexually intact male dogs were more numerous in the study population, castrated male dogs typically lived longer than spayed females or sexually intact males. Leading causes of death or euthanasia (76.3% of all dogs) were appendicular degenerative joint disease, neoplasia, spinal cord disease, nonspecific geriatric decline, and gastric dilatation-volvulus. Compared with German Shepherd Dogs, Belgian Shepherd Dogs were at increased risk for death attributable to neoplasia, behavior, and respiratory tract disease. German Shepherd Dogs had nearly twice the risk for death associated with spinal cord diseases, compared with Belgian Shepherd Dogs. CONCLUSIONS AND CLINICAL RELEVANCE: For most military working dogs, death or euthanasia results from a few diseases commonly associated with advanced age. Some breed differences in risk for these diseases may exist, which clinicians should consider in the procurement and long-term management of these dogs.     

Demonstration of Heterogeneous Genotypes of Taylorella equigenitalis Isolated from Horses in Six European Countries by Pulsed-Field Gel Electrophoresis

Forty-six isolates of Taylorella equigenitalis were analysed by pulsed-field gel electrophoresis (PFGE) after separate digestion of the genomic DNA with ApaI and with NotI. The isolates had been obtained from horses in six European countries and were classified into 18 genotypes. In Belgium, 2 genotypes were detected in 2 isolates, in England 9 among 15, in Finland 2 in 2, in France 2 among 10, in Sweden 3 among 5, and in Switzerland 3 among 12. Two English isolates and 4 French isolates gave identical PFGE profiles to those of Kentucky 188 from the United States. A common genotype was found in 5 isolates from Belgium and England and also in 10 isolates from France and Switzerland. The analysis of genomic DNA from 12 isolates of T. equigenitalis obtained from male horses in France, Sweden and Switzerland gave no evidence of a sex-related difference in the genomic DNA. Genomic DNA from 11 streptomycin (STM)-susceptible isolates obtained in Sweden and Switzerland were classified into four genotypes by PFGE. Each of the six genotypes determined among the 17 isolates from these two countries had single phenotypes for resistance or susceptibility to STM.