Accurate Measurement of the Optical Properties of Single Aerosol Particles Using Cavity Ring-Down Spectroscopy.

New approaches for the sensitive and accurate quantification of aerosol optical properties are needed to improve the current understanding of the unique physical chemistry of airborne particles and to explore their roles in fields as diverse as chemical manufacturing, healthcare, and atmospheric science. We have pioneered the use of cavity ring-down spectroscopy (CRDS), with concurrent angularly resolved elastic light scattering measurements, to interrogate the optical properties of single aerosol particles levitated in optical and electrodynamic traps. This approach enables the robust quantification of optical properties such as extinction cross sections for individual particles of known size. Our measurements can now distinguish the scattering and absorption contributions to the overall light extinction, from which the real and imaginary components of the complex refractive indices can be retrieved and linked to chemical composition. In this Feature Article, we show that this innovative measurement platform enables accurate and precise optical measurements for spherical and nonspherical particles, whether nonabsorbing or absorbing at the CRDS probe wavelength. We discuss the current limitations of our approach and the key challenges in physical and atmospheric chemistry that can now be addressed by CRDS measurements for single aerosol particles levitated in controlled environments.

Strategies for rapid rebreeding of lactating ewes in the spring.

Rapid rebreeding of winter- and spring-lambing ewes is essential if ewes are to lamb more than once per year, but fertility of lactating ewes is often low and early weaning of lambs may be undesirable in forage-based production systems. Selection to improve fertility in spring matings has been successful and led to development of ewes with a reduced seasonal anestrus. Potential for rapid rebreeding of lactating out-of-season breeding (OOS) ewes was tested in three studies. In Experiment 1, effects of short-term lamb removal on rebreeding was evaluated over 2 years using 71 January-lambing OOS ewes. At an average of 63 days postpartum, 36 ewes had lambs removed for 72 h, and all ewes were joined with rams. Circulating progesterone levels indicated that 74% of ewes ovulated before lamb separation; 91% of ewes mated within 5 weeks of ram exposure, 85% were diagnosed as pregnant and 75% lambed. The average interval between lambings was 225 days. In contrast to results observed in cattle, none of the measured variables was affected by lamb separation (P>0.20). Experiment 2 compared rebreeding performance of 24 OOS and 23 St. Croix ewes that lambed in January and averaged 60 days postpartum at ram introduction. More OOS ewes ovulated, mated and became pregnant during the first 21 days of ram exposure (83.3%, 58.3%, and 41.7%, respectively; P<0.001) compared with St. Croix ewes (26.1%, 0%, and 0%, respectively). After 39 days of ram exposure, pregnancy rates still favored OOS ewes (66.7% v. 39.1%; P=0.06), but the percentage of ewes that lambed did not differ (P>0.20) between OOS (47.8%) and St. Croix ewes (34.8%). In the third study, 34 March-lambing OOS ewes were exposed to rams on May 3 at an average of 40 days postpartum to characterize their reproductive performance. After 39 days of ram exposure, 52.9±8.7% of the ewes had mated, and 38.2±8.5% were diagnosed as pregnant. However, only 20.6±7.0% of the ewes produced viable lambs, suggesting a high level of uterine insufficiency. Spring fertility of lactating OOS ewes in these studies was one of the highest reported in the literature and indicated that selection for fertility in spring mating would improve reproductive performance in accelerated lambing programs. However, exposure of lactating OOS ewes to rams at 30 to 50 days postpartum was associated with high prenatal lamb mortality.

Ethics, politics and protests: using contentious issues in reproductive sciences as educational opportunities.

Contentious issues and polarized viewpoints can be utilized in the classroom and beyond to create a reflective dialogue among students and citizens. This dialogue leads to both a greater understanding, as well as an enhanced appreciation of alternative viewpoints. Exploring and discussing the scientific, ethical, moral, political, legal and societal aspects of contentious issues of human reproduction provides ideal subject matter for developing critical thinking skills in the field of reproductive science.

Effects of dietary supplementation with an organic source of selenium on characteristics of semen quality and in vitro fertility in boars.

Semen characteristics in boars fed organic or inorganic sources of Se were assessed in 3 experiments. Crossbred boars were randomly assigned at weaning to 1 of 3 dietary treatments: I) basal diets with no supplemental Se (control), II) basal diets with 0.3 mg/kg of supplemental Se from an organic source (Sel-Plex, Alltech Inc., Nicholasville, KY), and III) basal diets supplemented with 0.3 mg/kg of supplemental Se from sodium selenite (Premium Selenium 270, North American Nutrition Co. Inc., Lewisburg, OH). For Exp. 1, semen was collected from boars (n = 10/dietary treatment) on 5 consecutive days at 15 mo of age. Effects of treatment × day were detected for the proportions of progressively motile (P = 0.02) and rapidly moving (P = 0.03) spermatozoa, and measures of sperm velocity, including path velocity of the smoothed cell path (P = 0.05) and average velocity measured in a straight line from the beginning to the end of the track (P = 0.05). Negative effects of day of semen collection on sperm motility were least pronounced in boars fed Sel-Plex. Experiment 2 was conducted when boars were 17 mo of age, and semen was collected (n = 10 boars/dietary treatment), diluted in commercially available extenders, and stored at 18°C for 9 d. Effects of treatment × day were detected for percentages of motile (P = 0.01) and static (P = 0.01) spermatozoa, amplitude of lateral head displacement (P = 0.02), frequency with which the sperm track crossed the sperm path (P = 0.04), straightness (P = 0.01), and average size of all sperm heads (P = 0.03). In general, sperm cells from boars fed Sel-Plex were better able to maintain motility during liquid storage compared with boars fed sodium selenite. For Exp. 3, semen was collected from boars (n = 6/dietary treatment) at 23 mo of age, and spermatozoa were evaluated at d 1 and 8 after semen collection using in vitro fertilization procedures. There was a tendency for an effect (P = 0.11) of dietary treatment on fertilization rate with Sel-Plex-fed boars having the greatest value (70.7%). The results of this study suggest that there are positive effects of dietary supplementation with Sel-Plex on boar semen characteristics and that organic Se supplementation may help ameliorate the negative effects of semen storage on characteristics of sperm motility.

Effects of N-acetyl-cysteine and N-acetyl-cysteine-amide supplementation on in vitro matured porcine oocytes.

This study was conducted to evaluate the effects of different concentrations of the antioxidant N-acetyl-cysteine (NAC) supplemented to the maturation medium on porcine embryo development. Concentrations of NAC and its synthetic derivative, NAC-amide (NACA) were evaluated for effects on nuclear maturation, fertilization success and embryo development. Concentrations of NAC (0, 0.5, 1.0, 1.5, 2.0, 2.5 and 5.0 mm) were supplemented to maturing oocytes, and embryo development was analysed at 48 and 144 h post-fertilization. There were no differences among cleavage rates for any of the treatment groups. Blastocyst formation for 1.5 mm NAC (56.5 ± 9.2%) was higher (p < 0.05) than all other supplementations. There were no differences in nuclear maturation or fertilization or in cleavage rates when comparing 1.5 mm NAC and 1.5 mm NACA supplementation to the control. Blastocyst formation for 1.5 mm NAC (44.4 ± 4.7%) and 1.5 mm NACA (46.2 ± 3.4%) supplementation were higher (p < 0.05) than the control (32.1 ± 6.2%) oocytes. These results indicate that supplementing 1.5 mm of NAC or NACA to the oocyte maturation medium increased the percentage of viable embryos reaching the blastocyst stage of development.

Mechanisms of oxidative stress in porcine oocytes and the role of anti-oxidants.

The mechanisms of oxidative stress in in vitro maturing porcine oocytes and the effects of anti-oxidant supplementation of the medium in ameliorating these effects were investigated in the present study. In addition to intracellular reduced glutathione (GSH) concentrations and DNA fragmentation, the present study focused on superoxide dismutase (SOD), glutathione peroxidase (GPx) and catalase activity. The anti-oxidants used were N-acetylcysteine (NAC) and its derivative NAC-amide (NACA). The results indicate that when SOD is inhibited, supplementation of the maturarion medium with 1.5 mm NAC or NACA compensates for the decrease in SOD activity by reducing the degree of DNA fragmentation (P < 0.05). When GPx is inhibited, supplementation of the maturarion medium with 1.5 mm NAC alleviates the effects of no GPx activity, as indicated by a decrease in the degree of DNA fragmentation (P < 0.05). When the maturarion medium was supplemented with 1.5 mm NACA, intracellular GSH concentrations decreased (P < 0.05) and SOD and catalase activities increased (P < 0.05) along with the degree of DNA fragmentation. These results indicate that the mechanisms of alleviating oxidative stress in porcine oocytes are very complex and supplementing maturing oocytes with anti-oxidants may enhance enzyme activities and eliminate free radicals.

Effect of P.G. 600 on the timing of ovulation in gilts treated with altrenogest.

We previously reported that ovulation rate, but not pregnancy rate or litter size at d 30 after mating, was enhanced by treatment with P.G. 600 (400 IU of PMSG and 200 IU of hCG, Intervet America, Inc., Millsboro, DE) in gilts fed the orally active progestin, altrenogest (Matrix, Intervet America, Inc.) to synchronize estrus. We hypothesized that in addition to increasing ovulation rate, P.G. 600 may have altered the timing of ovulation. Therefore, mating gilts 12 and 24 h after first detection of estrus, as is common in the swine industry, may not have been the optimal breeding regimen, and as a consequence, pregnancy rate and litter size were not altered. The objective of the present study was to determine the effect of P.G. 600 on the timing of ovulation in gilts treated with altrenogest. Randomly cycling, crossbred gilts (5.5 mo old, 117 kg BW, and 14.7 mm of backfat) were fed a diet containing altrenogest (15 mg/d) for 18 d. Twenty-four hours after altrenogest withdrawal, gilts received i.m. injections of P.G. 600 (n = 25) or saline (n = 25). Gilts were checked for estrus at 8-h intervals. After first detection of estrus, transrectal ultrasonography was performed at 8-h intervals to determine the time of ovulation. Gilts were killed 9 to 11 d after the onset of estrus to determine ovulation rate. All gilts displayed estrus by 7 d after treatment with P.G. 600 or saline. Compared with saline, P.G. 600 increased (P = 0.07) ovulation rate (14.8 vs. 17.5, respectively; SE = 1.1). The intervals from injection to estrus (110.9 vs. 98.4; SE = 2.7 h; P < 0.01) and injection to ovulation (141.9 vs. 128.6; SE = 3.2 h; P < 0.01) were greater in gilts treated with saline than in gilts treated with P.G. 600. Duration of estrus (54.4 vs. 53.7; SE = 2.5 h), the estrus-to-ovulation interval (30.2 vs. 31.7; SE = 2.2 h), and the time of ovulation as a percentage of estrus duration (55.8 vs. 57.5; SE = 3.0%) did not differ for the P.G. 600 and saline-injected gilts, respectively. In summary, P.G. 600 advanced the onset of estrus and ovulation following termination of altrenogest treatment and increased ovulation rate; however, treatment of gilts with P.G. 600 had no effect on the timing of ovulation relative to the onset of estrus.

Plasma reduced folates, reproductive performance, and conceptus development in sows in response to supplementation with oxidized and reduced sources of folic acid.

The study was conducted to determine the response of sows to oxidized and reduced forms of supplemental folic acid in the diet. Gilts were mated and fed a standard corn-soybean meal diet with no supplemental folic acid. On d 105 of gestation, gilts were randomly assigned to one of four dietary treatments for the remainder of the study. Treatments were: 1) diet with no supplemental folate (control), 2) diet with 2.1 ppm (calculated) of added folate supplied by a synthetic pteroylmonoglutamate form (MG), 3) diet with 2.1 ppm (calculated) of added folate supplied by N5-formyl-5,6,7,8-tetrahydrofolic acid (THFA), or 4) a commercial bacterial cell powder source (Aj-PG) rich in reduced folates. Blood samples for high-performance liquid chromotography determination of reduced plasma folates were collected from gilts on d 105 of gestation, at weaning, at mating, and when the females were slaughtered on d 45 after mating for the second parity. There were 19, 18, 18, and 22 sows for the control, MG, THFA, and Aj-PG treatments, respectively. Supplementing folacin just before farrowing and during lactation had no effect on sow and litter performance during parity 1 (P > 0.10). Live fetuses at d 45 of gestation in Parity 2 were 10.06, 12.23, 10.87, and 11.07 for the control, MG, THFA, and Aj-PG treatments, respectively, and did not differ (P > 0.10). Fetal survival and placental size and protein content were generally unaffected by folate treatment. Concentration of reduced folates in sow plasma was 13.50, 13.58, 22.50, and 17.79 nM at weaning and 12.55, 19.29, 18.96, and 21.88 nM at mating for the control, MG, THFA, and Aj-PG treatments, respectively, with the THFA treatment elevated above the controls at weaning (P < 0.05) and the Aj-PG treatment greater than controls at mating (P < 0.05). At weaning, the reduced sources of supplemental folate (THFA and Aj-PG) were more effective in elevating plasma reduced folates than the oxidized folate supplement (MG; P < 0.05). Nonetheless, folate supplementation did not significantly improve sow reproductive performance in the subsequent parity, and there was no indication that reduced folate sources were superior to the oxidized pteroylmonoglutamate form as folate supplements for sows.

Serum and milk concentrations of leptin in gilts fed a high- or low-energy diet during gestation.

Concentrations of leptin in serum and milk were assessed in gilts fed diets during gestation that differed in energy level. Beginning at day 45 and continuing throughout pregnancy, gilts received either a high-energy (6882 kcal metabolizable energy (ME) per day) or low-energy (5221 kcal ME per day) diet (n = 9 per group). All gilts had ad libitum access to a standard lactation diet throughout a 21 day lactation. During gestation, gilts consuming the high-energy diet gained more body weight (P < 0.01) and backfat thickness (P = 0.03) than gilts fed the low-energy diet; however, serum concentrations of leptin remained similar between groups (P = 0.35). Within 24 h after farrowing, gilts fed the high-energy diet had greater levels of leptin in serum and milk than gilts that consumed the low-energy diet during gestation (P < 0.07); Across treatments, backfat thickness and leptin levels in serum were positively correlated (r(2) = 0.51; P = 0.03). At weaning, backfat thickness (P < 0.07), but not body weights or serum and milk levels of leptin (P > 0.1), were greater for gilts fed the high-energy, versus the low-energy, diet during gestation. Gilts that were fed the low-energy diet during gestation consumed more feed during week 2 of lactation (P = 0.06). Our results suggest that altering the level of energy in the diets of gestating swine can influence circulating and milk concentrations of leptin, as well as feed consumption, during lactation.

Temporal and geographic genetic variation in Culex pipiens quinquefasciatus (Diptera: Culicidae) from Florida.

Culex (Culex) pipiens quinquefasciatus Say field population from Vero Beach, FL, sampled monthly over a period of 8 mo, a colony sample, and six geographic samples were analyzed for genetic variation at 12 enzymes (10 "neutral" gene enzymes with 11 putative loci and two "complex" gene enzymes) by using polyacrylamide gel electrophoresis. The analysis of the 11 putative loci in both temporal and geographic samples showed that the four loci (Gpi, Hk, Mdhp-2, and Pgm) diagnostic of Cx. p. quinquefasciatus in the southern United States are present in similar frequencies in Florida samples. The Cx. p. quinquefasciatus colony sample showed significantly lower genetic variation than the temporal field samples, measured by mean number of alleles per locus (colony 1.2 +/- 0.1 versus field 1.44 +/- 0.03), percentage of polymorphic loci (colony 18.2% versus field 28.4%), mean observed heterozygosity (H(o) = colony 0.027 +/- 0.02 versus field 0.09 +/- 0.01), and mean Hardy-Weinberg expected heterozygosity (H(e) = colony 0.025 +/- 0.02 versus field 0.085 +/- 0.01). Three of the 11 loci (Acoh, Pgd, and Pgm) from the Vero Beach field samples showed bimodal patterns in their frequencies of the most common allele during peak density of the population. The low value of F(st) of 0.058 indicated minimum population substructuring among the temporal samples. Genetic variability values between geographic samples from the Florida panhandle and south Florida were not significant. Gene flow estimates based on F(ST), = 0.05, indicating low levels of gene flow among the geographic samples of Cx. p. quinquefasciatus. The average Nei's and modified Rogers' genetic distances among the six populations were 0.005 +/- 0.001 and 0.077 +/- 0.007, respectively. The cluster analysis did not suggest geographic clustering. The analysis of the "complex" gene enzymes in both temporal and geographic samples of Cx. p. quinquefasciatus from Florida showed the presence of two highly amplified esterases (Estbeta1 and Estalpha2\Estbeta2), indicating resistance to organophosphate insecticides and highly amplified Aldox enzyme (an enzyme that indicates resistance to at least one insecticide and a herbicide). Comparison of our results with previous studies on Cx. p. quinquefasciatus populations in the United States indicates that the genetic characteristics of the Florida populations of Cx. p. quinquefasciatus are very similar to populations from areas where ecological conditions are very different.

Temporal and geographic genetic variation in Culex nigripalpus theobald (Culicidae: Diptera), a vector of St. Louis encephalitis virus, from Florida.

A field population of Culex (Culex) nigripalpus Theobald from Vero Beach, FL sampled monthly over a period of 24 mo, a colony sample and 10 geographic samples were analyzed for genetic variation at 14 enzyme loci using polyacrylamide gel electrophoresis. The Cx. nigripalpus colony sample showed significantly lower genetic variation than the field-collected samples, measured by mean number of alleles per locus (colony 1.4 +/- 0.1 versus field 2.1 +/- 0.22), percentage of polymorphic loci (colony 35.7% versus field 54.8 +/- 7.7%), but mean observed heterozygosity (Ho = colony 0.16 +/- 0.07 versus field 0.17 +/- 0.03) and mean Hardy-Weinberg expected heterozygosity (He = colony 0.14 +/- 0.06 versus field 0.18 +/- 0.02) did not differ significantly. Three of the 14 loci (Aldox, Gpd, and Gpi) from the Vero Beach field samples showed distinct temporal patterns in the frequency of the most common allele. Higher mean observed heterozygosity (Ho) occurred during months following high rainfall in the Vero Beach field samples than during months following low rainfall. The average Nm value of 3.6 indicated high gene flow among the temporally distributed samples of the Vero Beach population. Genetic variability values between geographic samples from Panhandle, FL and south Florida were not significant. Gene flow estimates based on F(ST) = 0.039 provided a Nm of 6.2 indicating high levels of gene flow among the geographic samples of Cx. nigripalpus. The average Nei's and modified Rogers' genetic distances among the 10 populations were 0.009 +/- 0.001 and 0.081 +/- 0.004, respectively. The cluster analysis did not suggest geographic clustering, Because Cx. nigripalpus is the vector of St. Louis encephalitis (SLE) in Florida, temporal and geographic genetic variation in this species is discussed in relation to the seasonal and geographic SLE virus activity in Florida.

Isoenzyme variation in Aedes aegypti correlated with Dirofilaria immitis infectability.

From the Vero Beach strain of the mosquito Aedes (Stegomyia) aegypti (L.) (Diptera: Culicidae), substrains were selected for susceptibility (SS) and refractoriness (RR) to the dog heartworm Dirofilaria immitis (Leidy) (Filarioidea: Onchocercidae). These two lines and their reciprocal F1 hybrids were analysed for genetic variation at 14 enzyme loci, using polyacrylamide gel electrophoresis. Six of the enzyme loci showed variation (sample size 48 alleles/locus/line). Three of these were monomorphic in the refractory line but polymorphic in the susceptible, i.e. aconitase hydratase (Acoh), isocitrate dehydrogenase-1 (Idh-1) and phosphoglucomutase (Pgm). The other three loci, glucose-6-phosphate isomerase (Gpi), hexokinase-1 (Hk-1) and isocitrate dehydrogenase-2 (Idh-2), were polymorphic in both SS and RR lines and their hybrids. At two loci (Hk-1, Pgm) three alleles were detected, whereas the other polymorphic loci had only two alleles. For Hk-1, the most frequent allele was Hk-1(80) (0.563) in refractory and Hk-1(100) in the susceptible (0.521) and F1 hybrids. For Pgm the most frequent alleles were Pgm125 in the susceptible line (0.646) and Pgm100 in the F1 hybrids (0.563 and 0.604) and refractory line (1.000). The mean observed heterozygosity (Ho), the mean Hardy-Weinberg expected heterozygosity (He) and the mean number of alleles per locus in the refractory line were lower, but not significantly so, than in the susceptible line and their reciprocal F1 hybrids; the proportion of polymorphic loci was significantly lower in the refractory than in the susceptible line and their F1 hybrids. Within both lines all polymorphisms were in Hardy-Weinberg equilibrium, whereas significant departures from predicted frequencies were observed in SS x RR hybrids at four polymorphic loci (Acoh, Gpi, Hk-1, Pgm) and at three polymorphic loci (Acoh, Hk-1, Pgm) in RR x SS hybrids. The average Nei's and modified Rogers' genetic distances between the lines were 0.024 and 0.139, respectively. These electrophoretic data show that the refractory line (putatively lacking fi allele) can be distinguished from the susceptible line (fi/fi) and their hybrids (heterozygous fi) by isozyme marker frequencies, but it remains to be seen whether this difference is causal or chance linkage. In any case, this model system of Ae. aegypti/D. immitis provides opportunities to better understand and manipulate the molecular biology of filariasis transmission.

Effects of P.G. 600 on the onset of estrus and ovulation rate in gilts treated with Regu-mate.

Three experiments assessed the onset of estrus and ovulation rate in gilts treated with gonadotropins after the withdrawal of an orally active progestin. In Exp. 1, all cycling gilts received the progestin (Regu-mate; Intervet America Inc., Millsboro, DE) at a rate of 15 mg/d for 18 d. Twenty-four hours after the last feeding of Regu-mate, 32 gilts received an i.m. injection of 400 I.U. PMSG and 200 I.U. hCG (P.G. 600, Intervet America, Inc.), and 32 gilts received an i.m. injection of deionized water. The percentage of gilts displaying estrus < or = 7 d (P = 0.64) and the injection-to-estrus interval (P = 0.37) were similar for P.G. 600-treated gilts (93.8% and 4.1 +/- 0.1 d) and controls (90.6% and 4.3 +/- 0.1 d). Ovulation rate was greater (P < 0.01) in P.G. 600-treated gilts (28.8 +/- 1.1) compared with controls (17.4 +/- 1.1). In Exp. 2, 58 cycling gilts received Regu-mate (15 mg/d) for 18 d. Twenty-four hours after Regu-mate withdrawal, gilts received i.m. P.G. 600 or water (n = 29/treatment). Gilts were bred via AI 12 and 24 h after first detection of estrus. The percentage of gilts displaying estrus < or = 7 d (P = 0.45) and the injection-to-estrus interval (P = 0.27) were similar for P.G. 600-treated gilts (82.7% and 4.0 +/- 0.1 d) and controls (89.7% and 4.2 +/- 0.1 d). Ovulation rate was greater (P < 0.01) in P.G. 600-treated gilts (26.2 +/- 1.8) compared with controls (18.1 +/- 1.7). Pregnancy rate (P = 0.71) and the number of live embryos at d 30 postmating (P = 0.40) were similar for P.G. 600-treated gilts (91.7% and 15.6 +/- 1.2) and controls (88.5% and 14.1 +/- 1.2). In Exp. 3, prepubertal gilts (142.6 +/- 0.7 d of age) received Regumate (15 mg/d) (n = 20) or a control diet not including Regu-mate (n = 20) for 18 d. Twenty-four hours after Regu-mate withdrawal, all gilts received i.m. P.G. 600. The percentage of gilts displaying estrus < or = 7 d (P = 0.49) and the P.G. 600-to-estrus interval (P = 0.69) were similar for Regu-mate-fed gilts (95% and 4.3 +/- 0.2 d) and controls (88.9% and 4.2 +/- 0.2 d). Ovulation rate was similar (P = 0.38) for Regu-mate fed gilts (16.6 +/-1.6) and controls (14.4 +/- 1.8). In cycling gilts, administration of P.G. 600 after withdrawal of Regu-mate increased ovulation rate, but not litter size at d 30 postmating. There was no beneficial effect of Regu-mate pretreatment on the response to P.G. 600 in prepubertal gilts.

Aedes albopictus (Diptera: Culicidae): an experimental and natural host of Dirofilaria immitis (Filarioidea: Onchocercidae) in Florida, U.S.A.

Females of an Aedes albopictus (Skuse) colony from southeastern Florida, U.S.A., ingested low (22.9 +/- 3.2 mg/Female) and high (243.2 +/- 37.6 mf/Female) numbers of microfilariae from a dog infected with Dirofilaria immitis (Leidy). High mortality of females occurred during the first 4 d after infection regardless of the number of microfilariae ingested; daily mortality was almost negligible during 5-15 d after infection. Percentage of survival 15 d after infection was higher (63%) in females that ingested low numbers of microfilariae than those (15%) that ingested high numbers of microfilariae. The development of most of the D. immitis larvae was arrested in late L1 stage with some of the L1 stage larvae becoming melanized intracellularly in the Malpighian tubule cells of Ae. albopictus. Fifteen days after infection, development of D. immitis to the infective L3 stage occurred in only 10.9% of the surviving F1 and F2 Ae. albopictus that ingested low numbers of microfilariae, but in 94% of Anopheles quadrimaculatus Say ingesting similar numbers of microfilariae, as a control. Females of Ae. albopictus ingesting high numbers of microfilariae had more surviving females with L3 than those ingesting low numbers of microfilariae. The number of L3 larvae in the Malpighian tubules, hemocoel, head capsule and proboscis ranged from 1 to 37 per female, indicating the potential of Ae. albopictus to transmit D. immitis. Development of D. immitis larvae was not affected by co-infection with Ascogregarina taiwanensis (Lien & Levine), although both parasites infect the Malpighian tubules, the first intracellularly and the second extracellularly. After one generation of selection, a strain of Ae. albopictus susceptible to D. immitis developed 2.5 times more L3 than the parent strain. These results show that a small portion of the natural population of Ae. albopictus is susceptible to infection with D. immitis and that susceptibility may be increased rapidly by selection. The presence of developing Dirofilaria sp. larvae in the Malpighian tubules of field-caught females indicated that Ae. albopictus may be infected naturally with D. immitis in Florida.

Technical note: machine milking of sows--lactational milk yield and litter weights.

This study was designed to 1) determine milk yield of sows that were machine milked; 2) assess the effects of pulsation rate, pulsation ratio, and pig removal on milk yield; and 3) assess litter weights. In Exp. 1, four sows were milked daily to 60 d postpartum. There were differences (P < .05) in milk yield among sows, the greatest being 1,898 mL/d. Daily milk yield peaked between 15 and 25 d postpartum. Litter weights were 18.0 +/- 1.0 kg at farrowing and 60.8 +/- 12.1 kg at d 60. In Exp. 2, four sows were milked daily for 28 d. Pulsation rate and ratio (150/min and 28:72, milk:rest, and 60/min and 50:50) were alternated on a daily basis and pigs were isolated for either 0 or 60 min before milking. The higher pulsation rate resulted in more milk per milking (202 +/- 13 vs 168 +/- 13 mL; P < .05). Pig removal resulted in 221 +/- 11 vs 148 +/- 14 mL milk (P < .01). Pig removal times and pulsation characteristics affect the amount of milk obtained, but milk removal from sows has a severe effect on litter weight. This system can be used to harvest sow's milk for pharmaceutical purposes, but supplementation of the pigs is necessary.

Influence of pig substitution on milk yield, litter weights, and milk composition of machine milked sows.

This study was conducted to 1) determine milk yield of sows that were machine milked up to four times daily; 2) determine the effect of pig substitution on milk yield; 3) assess litter weight changes for sows that are milked; and 4) determine milk composition. Eight sows were milked four times daily to d 51 postpartum. Sows either maintained their own litter or had a week-old replacement litter to replace 25-d-old pigs. Individual gland milk yields were obtained on random days throughout lactation, and different diameter and weighted teat cups were rotated so that all glands received all combinations. Composite milk samples were analyzed for fat, protein, and somatic cells. Milk yields peaked at about 19 d postpartum and declined to 45 d postpartum in sows with their own litter, whereas milk yields peaked earlier and had a more dramatic decline after fostering of a younger litter. Litter weights were 17.1 +/- 1.0 kg at farrowing with 13.6 +/- .6 pigs born alive. Final litter weights were 34.4 +/- 11.7 kg for sows with replacement litters and 74.4 +/- 13.5 kg for sows with their own litters, and numbers of pigs weaned were 6.5 +/- 1.3 and 9.7 +/- 1.5, respectively. Milk fat was influenced by route of oxytocin administration (6.53 +/- .12 for i.v. vs 7.21 +/- .19% for i.m. administration; P < .05). Milk fat percentage was highest on d 2 and declined to 13 d postpartum. Milk protein was influenced by time of day of milking (lowest at the fourth milking, 5.57 +/- .11%) and followed a pattern similar to that for milk fat. Milk protein was affected in a linear manner by milk yield, with highest protein associated with lowest milk yields. Somatic cells in milk were influenced by litter replacement (P < .05) and oxytocin administration (P < .01). There was a linear increase in somatic cells from about 8 x 10(6) cells/mL milk at d 2 to more than 12 x 10(6) cells/mL milk at d 51 postpartum. These results show that pig replacement affects the amount of milk obtained. Moreover, milk composition changes throughout lactation. However, milk removal from sows has a severe impact on litter weight gains, and in systems where sow's milk is needed for commercial purposes, pig supplementation is necessary.

Laboratory evaluation of biotic and abiotic factors that may influence larvicidal activity of Bacillus thuringiensis serovar. israelensis against two Florida mosquito species.

A technical powder of Bacillus thuringiensis serovar. israelensis (B.t.i.) (VectoBac TP, 5,000 international toxic units [ITU]/mg), an aqueous suspension (VectoBac 12AS, 1,200 ITU/mg), and a granular formulation (VectoBac CG, 200 ITU/mg) were tested in the laboratory under different biotic and abiotic, conditions for efficacy against larvae of saltwater (Aedes taeniorhynchus) and freshwater (Culex nigripalpus) mosquitoes. Second-, 3rd-, and 4th-instar larvae of Cx. nigripalpus were 1.3-3-fold more susceptible to both VectoBac TP and VectoBac 12AS than were the respective larval instars of Ae. taeniorhynchus. For each species, 2nd-instar larvae were several-fold more susceptible to these B.t.i. preparations than were the 4th instars. In test cups, larvae under lower densities exposed to B.t.i. concentrations sustained 5-9-fold higher mortalities than larvae under high-density conditions. VectoBac TP and VectoBac 12AS stayed in suspension for up to 24 h with similar larvicidal efficacy, which was greater at 32-35 degrees C than at 15-20 degrees C. At 60 degrees C maintained for 24 h, VectoBac 12AS was adversely affected 2-3-fold in terms of potency, but VectoBac TP was not affected. Significant loss of potency of both VectoBac 12AS and VectoBac TP occurred when exposed to 35-37 degrees C under high light intensity (140,000-170,000 lux) for 6 h. Increasing salinity levels from 0 (fresh water) to 50% sea water caused gradual efficacy declines of VectoBac 12AS and VectoBac TP against Ae. taeniorhynchus larvae. VectoBac CG caused insignificant initial and residual (up to 8 days) larval mortalities of both mosquito species. This formulation did not release the active ingredient of B.t.i. in any significant mosquito larvicidal concentration in surface layers of water, and the formulation was more effective in shallower water. Storage of all 3 formulations under constant laboratory and variable field conditions for up to 8 months did not produce detectable potency loss of these products.

Colonization of Culex nigripalpus theobald (Diptera: Culicidae) by stimulation of mating using males of other mosquito species.

Culex nigripalpus Theobald (Diptera: Culicidae) was recolonized successfully by cohabiting Aedes taeniorhynchus males or males of other mosquito species starting in a large outdoor cage under natural light-dark cycles and temperatures and ending in a 1-ft.3 cage under artificial light-dark cycles at 24 degrees C without added stimulation.

Vector ability of mosquitoes for isolates of Plasmodium elongatum from raptors in Florida.

Three isolates of Plasmodium elongatum were obtained from 3 species of raptors (red-tailed hawk [Buteo jamaicensis], bald eagle [Haliaeetus leucocephalus], and eastern screech owl [Otus asio]) from Florida using isodiagnostic techniques in Pekin ducks (Anas platyrhynchos). Six to 10 species of mosquitoes were tested for susceptibility to these 3 isolates. Complete development of the sporogonic cycle of the 3 isolates of P. elongatum occurred in 3 species of mosquitoes, Culex nigripalpus, Culex restuans, and Culex salinarius. The pattern of susceptibility was similar among the 3 isolates of P. elongatum in Cx. nigripalpus. Culex restuans and Cx. salinarius were significantly more susceptible than Cx. nigripalpus to the 3 isolates of P. elongatum tested. Culex nigripalpus transmitted all 3 isolates of P. elongatum from duck to duck both by bite and after intraperitoneal injection of sporozoites. Infections of the 2 isolates tested occurred in ducks after intraperitoneal injection of sporozoites from Cx. restuans and Cx. salinarius. The results suggest that these 3 Culex species are potential vectors of P. elongatum from raptors in Florida.

Transgenic pigs produce functional human factor VIII in milk.

Deficiency or abnormality of coagulation factor VIII (FVIII) causes a bleeding disorder called hemophilia A. Treatment involves FVIII concentrates prepared from pooled human plasma or recombinant FVIII (rFVIII) prepared from mammalian cell culture. The cost of highly purified FVIII or rFVIII is a major factor in hemophilia therapy and restricts prophylaxis. We have sought to generate a new source of rFVIII by targeting expression of the human FVIII cDNA to the mammary gland of transgenic pigs using the regulatory sequences of the mouse whey acidic protein gene. The identity of processed heterodimeric rFVIII was confirmed using specific antibodies, by thrombin digestion and activity assays. The secretion of as much as 2.7 micrograms/ml of rFVIII in milk was over tenfold higher than in normal plasma. Up to 0.62 U/ml of rFVIII was detected in an assay in which rFVIII restored normal clotting activity to FVIII-deficient human plasma.